Patent Grant
7910606
The present invention is directed to a compound of formula I,
in which R0; R1; R2; R3; R4; Q; V, G and M have the meanings indicated below. The compound of formula I is a pharmacologically active compound. It exhibits a strong anti-thrombotic effect and is suitable, for example, for the therapy and prophylaxis of a cardiovascular disorder like a thromboembolic disease or restenosis. It is a reversible inhibitor of the blood clotting enzymes factor Xa (FXa) and/or factor VIIa (FVIIa), and can in general be applied in conditions in which an undesired activity of factor Xa and/or factor VIIa is present or for the cure or prevention of which an inhibition of factor Xa and/or factor VIIa is intended. The invention furthermore is directed to a process for the preparation of the compound of formula I, and pharmaceutical preparation comprising it.
Normal haemeostasis is the result of a complex balance between the processes of clot initiation, formation and clot dissolution. The complex interactions between blood cells, specific plasma proteins and the vascular surface, maintain the fluidity of blood unless injury and blood loss occurs (EP-A-987274). Many significant disease states are related to abnormal haemeostasis. For example, local thrombus formation due to rupture of atheroslerotic plaque is a major cause of acute myocardial infarction and unstable angina. Treatment of an occlusive coronary thrombus by either thrombolytic therapy or percutaneous angioplasty may be accompanied by acute thrombolytic reclosure of the affected vessel.
There continues to be a need for safe and effective therapeutic anticoagulants to limit or prevent thrombus formation. It is most desirable to develop agents that inhibit coagulation without directly inhibiting thrombin but by inhibiting other steps in the coagulation cascade like factor Xa and/or factor VIIa activity. It is now believed that inhibitors of factor Xa carry a lower bleeding risk than thrombin inhibitors (A. E. P. Adang & J. B. M. Rewinkel, Drugs of the Future 2000, 25, 369-383).
Low molecular weight, factor Xa-specific blood clotting inhibitors that are effective but do not cause unwanted side effects have been described, for example, in WO-A-95/29189. However, besides being an effective factor Xa-specific blood clotting inhibitor, it is desirable that such inhibitors also have further advantageous properties, for instance stability in plasma and liver and selectivity versus other serine proteases whose inhibition is not intended, such as thrombin. There is an ongoing need for further low molecular weight factor Xa specific blood clotting inhibitors, which are effective and have the above advantages as well.
Specific inhibition of the factor VIIa/tissue factor catalytic complex using monoclonal antibodies (WO-A-92/06711) or a protein such as chloromethyl ketone inactivated factor VIIa (WO-A-96/12800, WO-A-97/47651) is an extremely effective means of controlling thrombus formation caused by acute arterial injury or the thrombotic complications related to bacterial septicemia. There is also experimental evidence suggesting that inhibition of factor VIIa/tissue factor activity inhibits restenosis following balloon angioplasty. Bleeding studies have been conducted in baboons and indicate that inhibition of the factor VIIa/tissue factor complex has the widest safety window with respect to therapeutic effectiveness and bleeding risk of any anticoagulant approach tested including thrombin, platelet and factor Xa inhibition. Certain inhibitors of factor VIIa have already been described. EP-A-987274, for example discloses compounds containing a tripeptide unit, which inhibit factor VIIa. However, the property profile of these compounds is still not ideal, and there is an ongoing need for further low molecular weight factor VIIa inhibitory blood clotting inhibitors
In view of the current situation, it is clear that there is a need for a compound that exhibits better factor Xa and/or factor VIIa inhibitory activity and has high bioavailability.
The present invention is directed to a compound of formula I,
wherein
The invention also is directed to a method for preparing the compound according to the invention, a pharmaceutical preparation thereof, and its use for inhibiting factor Xa and/or factor VIIa or influencing blood coagulation or fibrinolysis.
The following are abbreviations used herein:
In general, the meaning of any group, residue, heteroatom, number etc., which can occur more than once in the compound of formulae I, Ib and Ic, is independent of the meaning of this group, residue, heteroatom, number etc. in any other occurrence. All groups, residues, heteroatoms, numbers etc., which can occur more than once in the compound of formulae I, Ib and Ic can be identical or different.
As used herein, the term alkyl is to be understood in the broadest sense to mean hydrocarbon residues which can be linear, i.e. straight-chain, or branched and which can be acyclic or cyclic residues or comprise any combination of acyclic and cyclic subunits. Further, the term alkyl as used herein expressly includes saturated groups as well as unsaturated groups which latter groups contain one or more, for example one, two or three, double bonds and/or triple bonds, provided that the double bonds are not located within a cyclic alkyl group in such a manner that an aromatic system results. All these statements also apply if an alkyl group occurs as a substituent on another residue, for example in an alkyloxy residue, an alkyloxycarbonyl residue or an arylalkyl residue. Examples of “—(C1-C8) -alkyl” or “—(C1-C8)-alkylene” are alkyl residues containing 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms are methyl, methylene, ethyl, ethylene, propyl, propylene, butyl, butylene, pentyl, pentylene, hexyl, heptyl or octyl, the n-isomers of all these residues, isopropyl, isobutyl, 1-methylbutyl, isopentyl, neopentyl, 2,2-dimethylbutyl, 2-methylpentyl, 3-methylpentyl, isohexyl, sec-butyl, t-Bu, tert-pentyl, sec-butyl, tert-butyl or tert-pentyl. The term “—(C0-C6) -alkyl” or “—(C0-C8)-alkylene” is a hydrocarbon residue containing 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms. The term “—C0-alkyl” or “—C0-alkylene” is a covalent bond.
Unsaturated alkyl residues are, for example, alkenyl residues such as vinyl, 1-propenyl, 2-propenyl (=allyl), 2-butenyl, 3-butenyl, 2-methyl-2-butenyl, 3-methyl-2-butenyl, 5-hexenyl or 1,3-pentadienyl, or alkynyl residues such as ethynyl, 1-propynyl, 2-propynyl (=propargyl) or 2-butynyl. Unsaturated alkyl residues can also be substituted.
Examples of —(C3-C8)-cycloalkyl cyclic alkyl residues are cycloalkyl residues containing 3, 4, 5, 6, 7 or 8 ring carbon atoms like cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyloheptyl or cyclooctyl, which can also be substituted and/or unsaturated. Unsaturated cyclic alkyl groups and unsaturated cycloalkyl groups like, for example, cyclopentenyl or cyclohexenyl can be bonded via any carbon atom.
Of course, a cyclic alkyl group has to contain at least three carbon atoms, and an unsaturated alkyl group has to contain at least two carbon atoms. Thus, a group like (C1-C8) -alkyl is to be understood as comprising, among others, saturated acyclic (C1-C8)-alkyl, (C3-C6)-cycloalkyl, and unsaturated (C2-C8)-alkyl like (C2-C8)-alkenyl or (C2-C8)-alkynyl. Similarly, a group like (C1-C4)-alkyl is to be understood as comprising, among others, saturated acyclic (C1-C4)-alkyl, and unsaturated (C2-C4)-alkyl like (C2-C4)-alkenyl or (C2-C4)-alkynyl.
Unless stated otherwise, the term alkyl preferably comprises acyclic saturated hydrocarbon residues which have from one to six carbon atoms and which can be linear or branched. A particular group of saturated acyclic alkyl residues is formed by (C1-C4)-alkyl residues like methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and t-Bu.
Unless stated otherwise, and irrespective of any specific substituents bonded to alkyl groups which are indicated in the definition of the compound of formulae I, Ib and Ic, alkyl groups can in general be unsubstituted or substituted by one or more, for example one, two or three, identical or different substituents. Any kind of substituents present in substituted alkyl residues can be present in any desired position provided that the substitution does not lead to an unstable molecule. Examples of substituted alkyl residues are alkyl residues in which one or more, for example 1, 2 or 3, hydrogen atoms are replaced with halogen atoms, in particular fluorine atoms.
The terms “a monocyclic or bicyclic 6- to 14-membered aryl” or “—(C6-C14)-aryl” are understood as meaning aromatic hydrocarbon radicals containing from 6 to 14 carbon atoms in the ring. Examples of —(C6-C14)-aryl radicals are phenyl, naphthyl, for example 1-naphthyl and 2-naphthyl, biphenylyl, for example 2-biphenylyl, 3-biphenylyl and 4-biphenylyl, anthryl or fluorenyl. Biphenylyl radicals, naphthyl radicals and, in particular, phenyl radicals are preferred aryl radicals.
The terms “mono- or bicyclic 4- to 15-membered heterocyclyl” or “—(C4-C15)-heterocyclyl” refer to heterocycles in which one or more of the 4 to 15 ring carbon atoms are replaced by heteroatoms such as nitrogen, oxygen or sulfur. Examples are acridinyl, azaindole (1H-pyrrolopyridinyl), azabenzimidazolyl, azaspirodecanyl, azepinyl, azetidinyl, aziridinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydrochinolinyl, 4,5-dihydrooxazolinyl, dioxazolyl, dioxazinyl, 1,3-dioxolanyl, 1,3-dioxolenyl, 6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]-tetrahydrofuranyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl (benzimidazolyl), isothiazolyl, isothiazolidinyl, isothiazolinyl, isoxazolyl, isoxazolinyl, isoxazolidinyl, 2-isoxazolinyl, ketopiperazinyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2-oxa-thiepanyl, 1,2-oxathiolanyl, 1,4-oxazepanyl, 1,4-oxazepinyl, 1,2-oxazinyl, 1,3-oxazinyl, 1,4-oxazinyl, oxazolidinyl, oxazolinyl, oxazolyl, oxetanyl, oxocanyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazolyl, pyridoimidazolyl, pyridothiazolyl, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolidinonyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydrothiophenyl, tetrazinyl, tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, 1,2-thiazinyl, 1,3-thiazinyl, 1,4-thiazinyl, 1,3-thiazolyl, thiazolyl, thiazolidinyl, thiazolinyl, thienyl, thietanyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thietanyl, thiomorpholinyl, thiophenolyl, thiophenyl, thiopyranyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5-triazinyl, 1,2,3-triazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl and xanthenyl.
Preferred are heterocyclyls, include benzimidazolyl, 1,3-benzodioxolyl, benzofuranyl, benzothiazolyl, benzothiophenyl, benzoxazolyl, chromanyl, cinnolinyl, 2-furyl, 3-furyl; imidazolyl, indolyl, indazolyl, isochromanyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, oxazolyl, phthalazinyl, pteridinyl, purinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridoimidazolyl, pyridopyridinyl, pyridopyrimidinyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrimidinyl, pyrrolyl; 2-pyrrolyl, 3-pyrrolyl, quinolinyl, quinazolinyl, quinoxalinyl, tetrazolyl, thiazolyl, 2-thienyl and 3-thienyl.
Also preferred are:
The terms “het” or “a 3- to 7-membered cyclic residue, containing up to 1, 2, 3 or 4 heteroatoms” refer to structures of heterocycles which can be derived from compounds such as azepine, azetidine, aziridine, azirine, 1,4 diazepane, 1,2-diazepine, 1,3-diazepine, 1,4-diazepine, diaziridine, diazirine, dioxazole, dioxazine, dioxole, 1,3-dioxolene, 1,3-dioxolane, furan, imidazole, imidazoline, imidazolidine, isothiazole, isothiazolidine, isothiazoline, isoxazole, isoxazoline, isoxazolidine,
The term “R1—N—R2—V can form a 4- to 8-membered cyclic group” or “R11 and R12 together with the nitrogen atom to which they are bonded can form a 4- to 8-membered monocyclic heterocyclic ring which in addition to the nitrogen atom can contain one or two identical or different ring heteroatoms chosen from oxygen, sulfur and nitrogen” refer to structures of heterocycles which can be derived from compounds such as azepane, azepine, azetidine, dioxazole, dioxazine, 1,4-diazepane, 1,2-diazepine, 1,3-diazepine, 1,4-diazepine, imidazole, imidazoline, imidazolidine, isothiazole, isothiazolidine, isothiazoline, isoxazole, isoxazoline, isoxazolidine, 2-isoxazoline, ketopiperazine, morpholine, [1,4]oxazepane, oxazole, piperazine, piperidine, pyrazine, pyrazole, pyrazoline, pyrazolidine, pyridazine, pyridine, pyrimidine, pyrrole, pyrrolidine, pyrrolidinone, pyrroline, tetrahydropyridine, tetrazine, tetrazole, thiazole, thiadiazole, thiazolidine, thiazoline, thiomorpholine, 1,2,3-triazine, 1,2,4-triazine, 1,3,5-triazine, 1,2,3-triazole or 1,2,4-triazole.
The term “R15 and R16 together with the carbon atom to which they are bonded can form a 3-to 6 membered carbocyclic ring” refer to structures, which can be derived from compounds such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
The term “R1 and R3 together with the atoms to which they are bonded can form a 6- to 8-membered cyclic group, containing up to 1, 2, 3 or 4 heteroatoms chosen from nitrogen, sulfur or oxygen” refers to structures of heterocycles which can be derived from compounds such as azocane, azocane-2-one, cyloheptyl cyclohexyl, cyclooctane, cyclooctene, 1,4-diazepane, 1,2-diazepine, 1,3-diazepine,
The fact that many of the before-listed names of heterocycles are the chemical names of unsaturated or aromatic ring systems does not imply that the, 4-15 membered mono- or polycyclic group could only be derived from the respective unsaturated ring system. The names here only serve to describe the ring system with respect to ring size and the number of the heteroatoms and their relative positions. As explained above, the 4-15 membered mono- or polycyclic group can be saturated or partially unsaturated or aromatic, and can thus be derived not only from the before-listed heterocycles themselves but also from all their partially or completely hydrogenated analogues and also from their more highly unsaturated analogues if applicable. As examples of completely or partially hydrogenated analogues of the before-listed heterocycles from which this group may be derived the following may be mentioned: pyrroline, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, dihydropyridine, tetrahydropyridine, piperidine, 1,3-dioxolane, 2-imidazoline, imidazolidine, 4,5-dihydro-1,3-oxazole, 1,3-oxazolidine, 4,5-dihydro-1,3-thiazole, 1,3-thiazolidine, perhydro-1,4-dioxane, piperazine, perhydro-1,4-oxazine (=morpholine), perhydro-1,4-thiazine (=thiomorpholine), perhydroazepine, indoline, isoindoline, 1,2,3,4-tetrahydroquinoline, 1,2,3,4-tetrahydroisoquinoline, etc.
The 4-15 membered mono- or polycyclic group may be bonded via any ring carbon atom, and in the case of nitrogen heterocycles via any suitable ring nitrogen atom. Thus, for example, a pyrrolyl residue can be 1-pyrrolyl, 2-pyrrolyl or 3-pyrrolyl, a pyrrolidinyl residue can be pyrrolidin-1-yl
Unless stated otherwise, and irrespective of any specific substituents bonded to the 4-15 membered mono- or polycyclic group or any other heterocyclic groups which are indicated in the definition of the compound of formulae I, Ib and Ic, the 4-15 membered mono- or polycyclic group can be unsubstituted or substituted on ring carbon atoms with one or more, for example one, two, three, four or five, identical or different substituents like (C1-C8)-alkyl, in particular (C1-C4)-alkyl, (C1-C8)-alkyloxy, in particular (C1-C4)-alkyloxy, (C1-C4)-alkylthio, halogen, nitro, amino, ((C1-C4) -alkyl)carbonylamino like acetylamino, trifluoromethyl, trifluoromethoxy, hydroxy, oxo, hydroxy-(C1-C4)-alkyl such as, for example, hydroxymethyl or 1-hydroxyethyl or 2-hydroxyethyl, methylenedioxy, ethylenedioxy, formyl, acetyl, cyano, aminosulfonyl, methylsulfonyl, hydroxycarbonyl, aminocarbonyl, (C1-C4)-alkyloxycarbonyl, optionally substituted phenyl, optionally substituted phenoxy, benzyl optionally substituted in the phenyl group, benzyloxy optionally substituted in the phenyl group, etc. The substituents can be present in any desired position provided that a stable molecule results. Of course an oxo group cannot be present in an aromatic ring. Each suitable ring nitrogen atom in the 4-15 membered mono- or polycyclic group can independently of each other be unsubstituted, i.e. carry a hydrogen atom, or can be substituted, i.e. carry a substituent like (C1-C8)-alkyl, for example (C1-C4)-alkyl such as methyl or ethyl, optionally substituted phenyl, phenyl-(C1-C4)-alkyl, for example benzyl, optionally substituted in the phenyl group, hydroxy-(C2-C4)-alkyl such as, for example 2-hydroxyethyl, acetyl or another acyl group, methylsulfonyl or another sulfonyl group, aminocarbonyl, (C1-C4)-alkyloxycarbonyl, etc. In general, in the compound of formulae I, Ib and Ic a nitrogen heterocycle can also be present as N-oxides or as quaternary salts. Ring sulfur atoms can be oxidized to the sulfoxide or to the sulfone. Thus, for example a tetrahydrothienyl residue may be present as S,S-dioxotetrahydro-thienyl residue or a thiomorpholinyl residue like thiomorpholin-4-yl may be present as 1-oxo -thiomorpholin-4-yl or 1,1-dioxo-thiomorpholin-4-yl. A substituted 4 to 15 membered mono- or polycyclic group that can be present in a specific position of the compounds of formulae I, Ib and Ic can independently of other groups be substituted by substituents selected from any desired subgroup of the substituents listed before and/or in the definition of that group.
The 3-7 membered monocyclic group may be bonded via any ring carbon atom, and in the case of nitrogen heterocycles via any suitable ring nitrogen atom. Thus, for example, a pyrrolyl residue can be 1-pyrrolyl, 2-pyrrolyl or 3-pyrrolyl, a pyrrolidinyl residue can be pyrrolidin-1-yl (=pyrrolidino), pyrrolidin-2-yl or pyrrolidin-3-yl, a pyridinyl residue can be pyridin-2-yl, pyridin-3-yl or pyridin-4-yl, a piperidinyl residue can be piperidin-1-yl (=piperidino), piperidin-2-yl, piperidin-3-yl or piperidin-4-yl. Furyl can be 2-furyl or 3-furyl, thienyl can be 2-thienyl or 3-thienyl, imidazolyl can be imidazol-1-yl, imidazol-2-yl, imidazol-4-yl or imidazol-5-yl, 1,3-oxazolyl can be 1,3-oxazol-2-yl, 1,3-oxazol-4-yl or 1,3-oxazol-5-yl, 1,3-thiazolyl can be 1,3-thiazol-2-yl, 1,3-thiazol-4-yl or 1,3-thiazol-5-yl, pyrimidinyl can be pyrimidin-2-yl, pyrimidin-4-yl (=6-pyrimidinyl) or 5-pyrimidinyl, piperazinyl can be piperazin-1-yl (=piperazin-4-yl=piperazino) or piperazin-2-yl. Unless stated otherwise, and irrespective of any specific substituents bonded to the 3-7 membered monocyclic group or any other heterocyclic groups which are indicated in the definition of the compound of formulae I, Ib and Ic, can be unsubstituted or substituted on ring carbon atoms with one or more, for example one, two, three, four or five, identical or different substituents like (C1-C8)-alkyl, in particular (C1-C4)-alkyl, (C1-C8)-alkyloxy, in particular (C1-C4)-alkyloxy, (C1-C4)-alkylthio, halogen, nitro, amino, ((C1-C4)-alkyl)carbonylamino like acetylamino, trifluoromethyl, trifluoromethoxy, hydroxy, oxo, hydroxy-(C1-C4)-alkyl such as, for example, hydroxymethyl or 1-hydroxyethyl or 2-hydroxyethyl, methylenedioxy, ethylenedioxy, formyl, acetyl, cyano, aminosulfonyl, methylsulfonyl, hydroxycarbonyl, aminocarbonyl, (C1-C4)-alkyloxycarbonyl, optionally substituted phenyl, optionally substituted phenoxy, benzyl optionally substituted in the phenyl group, benzyloxy optionally substituted in the phenyl group, etc. The substituents can be present in any desired position provided that a stable molecule results. Of course an oxo group cannot be present in an aromatic ring. Each suitable ring nitrogen atom in the 3-7 membered monocyclic group can independently of each other be unsubstituted, i.e. carry a hydrogen atom, or can be substituted, i.e. carry a substituent like (C1-C8)-alkyl, for example (C1-C4)-alkyl such as methyl or ethyl, optionally substituted phenyl, phenyl-(C1-C4)-alkyl, for example benzyl, optionally substituted in the phenyl group, hydroxy-(C2-C4)-alkyl such as, for example 2-hydroxyethyl, acetyl or another acyl group, methylsulfonyl or another sulfonyl group, aminocarbonyl, (C1-C4)-alkyloxycarbonyl, etc. In general, in the compound of formulae I nitrogen heterocycles can also be present as N-oxides or as quaternary salts. Ring sulfur atoms can be oxidized to the sulfoxide or to the sulfone. Thus, for example a tetrahydrothienyl residue may be present as S,S-dioxotetrahydrothienyl residue or a thiomorpholinyl residue like thiomorpholin-4-yl may be present as 1-oxo-thiomorpholin-4-yl or 1,1-dioxo-thiomorpholin-4-yl. A substituted 3-7 membered monocyclic group that can be present in a specific position of the compounds of formulae I can independently of other groups be substituted by substituents selected from any desired subgroup of the substituents listed before and/or in the definition of that group.
The term “—(C1-C3)-perfluoroalkyl” is a partial or totally fluorinated alkyl-residue, which can be derived from residues such as —CF3, —CHF2, —CH2F, —CHF—CF3, —CHF—CHF2, —CHF—CH2F, —CH2—CF3, —CH2—CHF2, —CH2—CH2F, —CF2—CF3, —CF2—CHF2, —CF2—CH2F, —CH2—CHF—CF3, —CH2—CHF—CHF2, —CH2—CHF—CH2F, —CH2—CH2—CF3, —CH2—CH2—CHF2, —CH2—CH2—CH2F, —CH2—CF2—CF3, —CH2—CF2—CHF2, —CH2—CF2—CH2F, —CHF—CHF—CF3, —CHF—CHF—CHF2, —CHF—CHF—CH2F, —CHF—CH2—CF3, —CHF—CH2—CHF2, —CHF—CH2—CH2F, —CHF—CF2—CF3, —CHF—CF2—CHF2, —CHF—CF2—CH2F, —CF2—CHF—CF3, —CF2—CHF—CHF2, —CF2—CHF—CH2F, —CF2—CH2—CF3, —CF2—CH2—CHF2, —CF2—CH2—CH2F, —CF2—CF2—CF3, —CF2—CF2—CHF2 or —CF2—CF2—CH2F.
The term “—(C1-C3)-perfluoroalkylene” is a partial or totally fluorinated alkylene-residue, which can be derived from residues such as —CF2—, —CHF—, —CHF—CHF2—, —CHF—CHF—, —CH2—CF2—, —CH2—CHF—, —CF2—CF2—, —CF2—CHF—, —CH2—CHF—CF2—, —CH2—CHF—CHF—, —CH2—CH2—CF2—, —CH2—CH2—CHF, —CH2—CF2—CF2—, —CH2—CF2—CHF—, —CHF—CHF—CF2—, —CHF—CHF—CHF—, —CHF—CH2—CF2—, —CHF—CH2—CHF—, —CHF—CF2—CF2—, —CHF—CF2—CHF—, —CF2—CHF—CF2—, —CF2—CHF—CHF—, —CF2—CH2—CF2—, —CF2—CH2—CHF—, —CF2—CF2—CF2—, or —CF2—CF2—CHF.
Halogen is fluorine, chlorine, bromine or iodine, preferably fluorine, chlorine or iodune, particularly preferably chlorine or iodine.
Optically active carbon atoms present in the compound of formulae I, Ib and Ic can independently of each other have R configuration or S configuration. the compound of formulae I, Ib and Ic can be present in the form of pure enantiomers or pure diastereomers or in the form of mixtures of enantiomers and/or diastereomers, for example in the form of racemates. The present invention is directed to pure enantiomers and mixtures of enantiomers as well as to pure diastereomers and mixtures of diastereomers. The invention comprises mixtures of two or of more than two stereoisomers of the formulae I, Ib and Ic, and it comprises all ratios of the stereoisomers in the mixtures. In case the compound of formulae I, Ib and Ic can be present as E isomers or Z isomers (or cis isomers or trans isomers) the invention relates both to pure E isomers and pure Z isomers and to E/Z mixtures in all ratios. The invention also comprises all tautomeric forms of the compound of formulae I, Ib and Ic.
Diastereomers, including E/Z isomers, can be separated into the individual isomers, for example, by chromatography. Racemates can be separated into the two enantiomers by customary methods, for example by chromatography on chiral phases or by resolution, for example by crystallization of diastereomeric salts obtained with optically active acids or bases. Stereochemically uniform a compound of formulae I, Ib and Ic can also be obtained by employing stereochemically uniform starting materials or by using stereoselective reactions. Physiologically tolerable salts of the compounds of formulae I, Ib and Ic are nontoxic salts that are physiologically acceptable, in particular pharmaceutically utilizable salts. Such salts of a compound of formulae I, Ib and Ic containing acidic groups, for example a carboxyl group COOH, are for example alkali metal salts or alkaline earth metal salts such as sodium salts, potassium salts, magnesium salts and calcium salts, and also salts with physiologically tolerable quaternary ammonium ions such as tetramethylammonium or tetraethylammonium, and acid addition salts with ammonia and physiologically tolerable organic amines, such as methylamine, dimethylamine, trimethylamine, ethylamine, triethylamine, ethanolamine or tris-(2-hydroxyethyl)amine. Basic groups contained in the compound of formulae I, Ib and Ic, for example amino groups or guanidino groups, form acid addition salts, for example with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid or phosphoric acid, or with organic carboxylic acids and sulfonic acids such as formic acid, acetic acid, oxalic acid, citric acid, lactic acid, malic acid, succinic acid, malonic acid, benzoic acid, maleic acid, fumaric acid, tartaric acid, methanesulfonic acid or p-toluenesulfonic acid. A compound of formulae I, Ib and Ic, which simultaneously contain a basic group and an acidic group, for example a guanidino group and a carboxyl group, can also be present as zwitterions (betaines) which are likewise included in the present invention.
Salts of a compound of formulae I, Ib and Ic can be obtained by customary methods known to those skilled in the art, for example by combining a compound of the formulae I, Ib and Ic with an inorganic or organic acid or base in a solvent or dispersant, or from other salts by cation exchange or anion exchange. The present invention also includes all salts of the compound of formulae I, Ib and Ic which, because of low physiologically tolerability, are not directly suitable for use in pharmaceuticals but are suitable, for example, as intermediates for carrying out further chemical modifications of the compound of formulae I, Ib and Ic or as starting materials for the preparation of physiologically tolerable salts.
The present invention furthermore includes all solvates of a compound of formulae I, Ib and Ic, for example hydrates or adducts with alcohols.
The invention also includes derivatives and modifications of the compound of formulae I, Ib and Ic, for example prodrugs, protected forms and other physiologically tolerable derivatives, as well as active metabolites of the compound of formulae I, Ib and Ic. The invention relates in particular to prodrugs and protected forms of the compound of formulae I, Ib and Ic, which can be converted into a compound of formulae I, Ib and Ic under physiological conditions. Suitable prodrugs for the compound of formulae I, Ib and Ic, i.e. chemically modified derivatives of the compound of formulae I, Ib and Ic having properties which are improved in a desired manner, for example with respect to solubility, bioavailability or duration of action, are known to those skilled in the art. More detailed information relating to prodrugs is found in standard literature like, for example, Design of Prodrugs, H. Bundgaard (ed.), Elsevier, 1985; Fleisher et al., Advanced Drug Delivery Reviews 19 (1996) 115-130; or H. Bundgaard, Drugs of the Future 16 (1991) 443 which are all incorporated herein by reference. Suitable prodrugs for the compound of formulae I, Ib and Ic are especially acyl prodrugs and carbamate prodrugs of acylatable nitrogen-containing groups such as amino groups and the guanidino group and also ester prodrugs and amide prodrugs of carboxylic acid groups which may be present in a compound of formulae I, Ib and Ic. In the acyl prodrugs and carbamate prodrugs one or more, for example one or two, hydrogen atoms on nitrogen atoms in such groups are replaced with an acyl group or a carbamate, preferably a —(C1-C6)-alkyloxycarbonyl group. Suitable acyl groups and carbamate groups for acyl prodrugs and carbamate prodrugs are, for example, the groups Rp1—CO— and Rp2O—CO—, in which Rp1 is hydrogen, (C1-C18)-alkyl, (C3-C8)-cycloalkyl, (C3-C8)-cycloalkyl-(C1-C4)-alkyl-, (C6-C14)-aryl, Het-, (C6-C14)-aryl-(C1-C4)-alkyl- or Het-(C1-C4)-alkyl- and in which Rp2 has the meanings indicated for Rp1 with the exception of hydrogen.
Especially preferred a compound of formulae I, Ib and Ic are those wherein two or more residues are defined as indicated before for preferred a compound of formulae I, Ib and Ic, or residues can have one or some of the specific denotations of the residues given in their general definitions or in the definitions of preferred compounds before. All possible combinations of definitions given for preferred definitions and of specific denotations of residues explicitly are a subject of the present invention.
A particular embodiment of the compound of formula I according to the invention is wherein
Another particular embodiment of the compound of formula I according to the invention is wherein
Another particular embodiment of the compound of formula I according to the invention is wherein
Another particular embodiment of the compound of formula I according to the invention is wherein
Another particular embodiment of the compound of formula I according to the invention is wherein
Another particular embodiment of the compound of formula I according to the invention is wherein
Another particular embodiment of the compound of formula I according to the invention is wherein
Another particular embodiment of the compound of formula I according to the invention is wherein
Another particular embodiment of the compound of formula I according to the invention is wherein
Another particular embodiment of the compound of formula I according to the invention is wherein
Another particular embodiment of the invention is wherein the compound of formula I has the following formula
Another particular embodiment of the invention is wherein the compound of formula I has the following formula
Another particular embodiment of the invention is wherein the compound of formula I is a species selected from the group consisting of
Another particular embodiment of the invention is a process for the preparation of the compound according to claim 1, comprising condensing a carboxylic acid of the formula 2
Another particular embodiment of the invention is a pharmaceutical preparation comprising a pharmaceutically effective amount of at least one compound according to claim 1 and a pharmaceutically acceptable carrier.
Another particular embodiment of the invention is wherein a method of inhibiting the activity of factor Xa or factor VIIa comprising contacting an inhibitory amount of a compound according to claim 1 with a composition containing factor Xa or factor VIIa to influence blood coagulation.
Another particular embodiment of the invention is wherein the blood coagulation is connected with abnormal thrombus formation, acute myocardial infarction, cardiovascular disorders, unstable angina, thromboembolism, acute vessel closure associated with thrombolytic therapy or percutaneous transluminal coronary angioplasty (PTCA), transient ischemic attacks, stroke, intermittent claudication, bypass grafting of the coronary or peripheral arteries, vessel luminal narrowing, restenosis post coronary or venous angioplasty, maintenance of vascular access patency in long-term hemodialysis patients, pathologic thrombus formation occurring in the veins of the lower extremities following abdominal, knee or hip surgery, pathologic thrombus formation occurring in the veins of the lower extremities following abdominal, knee and hip surgery, a risk of pulmonary thromboembolism, or disseminated systemic intravascular coagulatopathy occurring in vascular systems during septic shock, viral infections or cancer, or reducing an inflammatory response, fibrinolysis, or treatment of coronary heart disease, myocardial infarction, angina pectoris, vascular restenosis, adult respiratory distress syndrome, multi-organ failure and disseminated intravascular clotting disorder, deep vein or proximal vein thrombosis.
Another particular embodiment of the invention is wherein a method of inhibiting the activity of factor Xa or factor VIIa comprising contacting an inhibitory amount of a compound according to claim 1 with a composition containing factor Xa or factor VIIa to influence fibrinolysis.
Also with respect to all preferred a compound of formulae I, Ib and Ic all their stereoisomeric forms and mixtures thereof in any ratio and their physiologically acceptable salts explicitly are a subject of the present invention, as well as are their prodrugs. Similarly, also in all preferred a compound of formulae I, Ib and Ic, all residues that are present more than one time in the molecule are independent of each other and can be identical or different.
The compound of formulae I, Ib and Ic can be prepared by utilising procedures and techniques, which per se are well known and appreciated by one of ordinary skill in the art. Starting materials or building blocks for use in the general synthetic procedures that can be applied in the preparation of the compounds of formulae I, Ib and Ic are readily available to one of ordinary skill in the art. In many cases they are commercially available or have been described in the literature. Otherwise they can be prepared from readily available precursor compounds analogously to procedures described in the literature, or by procedures or analogously to procedures described in this application.
In general, a compound of formulae I, Ib and Ic can be prepared, for example in the course of a convergent synthesis, by linking two or more fragments which can be derived retrosynthetically from the formulae I, Ib and Ic. More specifically, suitably substituted starting Pyrazole derivatives are employed as building blocks in the preparation of the compounds of formulae I, Ib and Ic. If not commercially available, such Pyrazole derivatives can be prepared according to the well-known standard procedures for the formation of the Pyrazole ring system. By choosing suitable precursor molecules, these pyrazole syntheses allow the introduction of a variety of substituents into the various positions of the pyrazole system, which can be chemically modified in order to finally arrive at the molecule of the formulae I, Ib and Ic having the desired substituent pattern. As one of the comprehensive reviews in which numerous details and literature references on the chemistry of pyrazole and on synthetic procedures for their preparation can be found, J. Eiguero in “Comprehensive Heterocyclic Chemistry II”; Eds. A. Katritzky, Ch. Rees, E. Scriven; Elsevier 1996, Vol. 3; K. Kirschke in Houben-Weyl, “Methoden der Organischen Chemie” (Methods of Organic Chemistry), Georg Thieme Verlag, Stuttgart, Germany 1994, Vol. E8b Hetarene; T. Nagai et al. Org. Prep. Proced. Int. (1993), 25, 403; M. Elnagdi et al. Heterocycles (1985) 23, 3121; K. Makino et al. J. Heterocycl. Chem. (1998) 35, 489; K. Makino et al. J. Heterocycl. Chem. (1999) 36, 321.
If starting pyrazole derivatives are not commercially available and have to be synthesized this can be done, for example, according to the well-known pyrazole syntheses mentioned above. In the following procedures of particuluar interest for the embodiment of this invention are listed and referenced briefly, however, they are standard procedures comprehensively discussed in the literature, and are well known to one skilled in the art. Although not always shown explicitly, in certain cases positional isomers will occur during the synthesis of the below mentioned reactions. Nevertheless such mixtures of positional isomers, can be separated by modern separation techniques like, for example, preparative HPLC.
Further, in order to obtain the desired substituents at the pyrazole ring system in the formulae I, Ib and Ic, the functional groups introduced into the ring system during the pyrazole synthesis can be chemically modified. Especially the groups present in the pyrazole ring system can be modified by a variety of reactions and thus the desired residues R1
Depending on the reaction conditions, reagent, stochiometry and substitution pattern the halogen is introduced in the 3-position and/or 4-position and/or 5-position. By selective halogen/metal exchange or metalation by selective hydrogen/metal exchange and subsequent reaction with a wide range of electrophiles various substituents can be introduced at the heterocyclic nucleus. (M. R. Grimmett, Heterocycles (1994) 37, 2087; V. D. Gardner et al., J. Heterocycl. Chem. (1984), 21, 121; D. Butler et al., J. Org. Chem. (1971) 36, 2542). Halogens or hydroxy groups (via their triflates or nonaflates)—or primary amines (via their diazonium salts) present in the pyrazole structure—can be converted directly, or after interconversion to the corresponding stannane, or boronic acid, into a variety of other functional groups like for example —CN, —CF3, —C2F5, ethers, acids, amides, amines, alkyl- or aryl-groups mediated by means of transition metals, namely palladium or nickel catalysts or copper salts and reagents for example referred to below (F. Diederich, P. Stang, Metal-catalyzed Cross-coupling Reactions, Wiley-VCH, 1998; or M. Beller, C. Bolm, Transition Metals for Organic Synthesis, Wiley-VCH, 1998; J. Tsuji, Palladium Reagents and Catalysts, Wiley, 1996; J. Hartwig, Angew. Chem. (1998) 110, 2154; B. Yang, S. Buchwald, J. Organomet. Chem. (1999) 576, 125; T. Sakamoto, K. Ohsawa, J. Chem. Soc. Perkin Trans I (1999) 2323; D. Nichols, S. Frescas, D. Marona-Lewicka, X. Huang, B. Roth, G. Gudelsky, J. Nash, J. Med. Chem. (1994) 37, 4347; P. Lam, C. Clark, S. Saubem, J. Adams, M. Winters, D. Chan, A. Combs, Tetrahedron Lett. (1998) 39, 2941; D. Chan, K. Monaco, R. Wang, M. Winters, Tetrahedron Lett. (1998) 39, 2933; V. Farina, V. Krishnamurthy, W. Scott, The Stille Reaction, Wiley, 1994; F. Qing et al. J. Chem. Soc. Perkin Trans. 1 (1997) 3053; S. Buchwald et al. J. Am. Chem. Soc. (2001) 123, 7727; S. Kang et al. Synlett (2002) 3, 427; S. Buchwald et al. Organic Lett. (2002) 4, 581; T. Fuchikami et al. Tetrahedron Lett. (1991) 32, 91; Q. Chen et al. Tetrahedron Lett. (1991) 32, 7689).
Nitro groups can be reduced to amino groups by means of various reducing agents, such as sulfides, dithionites, complex hydrides or by catalytic hydrogenation. A reduction of a nitro group may also be carried out at a later stage of the synthesis of a compound of the formulae I, Ib and Ic, and a reduction of a nitro group to an amino group may also occur simultaneously with a reaction performed on another functional group, for example when reacting a group like a cyano group with hydrogen sulfide or when hydrogenating a group. In order to introduce the residues R1
Ester groups present in the pyrazole nucleus can be hydrolyzed to the corresponding carboxylic acids, which after activation can then be reacted with amines or alcohols under standard conditions to give amides or alcohols, respectively. Ester groups present in the pyrazole nucleus can be converted to other esters by transesterification. Carboxylic acids attached to a suitable pyrazole nucleus can also be alkylated to give esters. Ether groups present at the pyrazole nucleus, for example benzyloxy groups or other easily cleavable ether groups, can be cleaved to give hydroxy groups which then can be reacted with a variety of agents, for example etherification agents or activating agents allowing replacement of the hydroxy group by other groups. Sulfur-containing groups can be reacted analogously.
During the course of the synthesis in order to modify the groups R87 or R8′ attached to the pyrazole ring system by application of parallel synthesis methodology, a variety of reactions can be extremely useful, including, for example, palladium, nickel or copper catalysis. Such reactions are described for example in F. Diederich, P. Stang, Metal-catalyzed Cross-coupling Reactions, Wiley-VCH (1998); or M. Beller, C. Bolm, Transition Metals for Organic Synthesis, Wiley-VCH (1998); J. Tsuji, Palladium Reagents and Catalysts, Wiley (1996); J. Hartwig, Angew. Chem. (1998), 110, 2154; B. Yang, S. Buchwald, J. Organomet. Chem. (1999), 576, 125; P. Lam, C. Clark, S. Saubern, J. Adams, M. Winters, D. Chan, A. Combs, Tetrahedron Lett. (1998), 39, 2941; D. Chan, K. Monaco, R. Wang, M. Winters, Tetrahedron Lett. (1998), 39, 2933; J. Wolfe, H. Tomori, J. Sadight, J. Yin, S. Buchwald, J. Org. Chem. (2000), 65, 1158; V. Farina, V. Krishnamurthy, W. Scott, The Stille Reaction, Wiley, (1994); S. Buchwald et al., J. Am. Chem. Soc. (2001), 123, 7727; S. Kang et al., Synlett (2002), 3, 427; S. Buchwald et al., Org. Lett. (2002), 4, 581.
The previously-mentioned reactions for the conversion of functional groups are furthermore, in general, extensively described in textbooks of organic chemistry like M. Smith, J. March, March's Advanced Organic Chemistry, Wiley-VCH, 2001 and in treatises like Houben-Weyl, “Methoden der Organischen Chemie” (Methods of Organic Chemistry), Georg Thieme Verlag, Stuttgart, Germany, or “Organic Reactions”, John Wiley & Sons, New York, or R. C. Larock, “Comprehensive Organic Transformations”, Wiley-VCH, 2nd ed (1999), B. Trost, I. Fleming (eds.) Comprehensive Organic Synthesis, Pergamon, 1991; A. Katritzky, C. Rees, E. Scriven Comprehensive Heterocyclic Chemistry II, Elsevier Science, 1996) in which details on the reactions and primary source literature can be found. Due to the fact that in the present case the functional groups are attached to an pyrazole ring it may in certain cases become necessary to specifically adapt reaction conditions or to choose specific reagents from a variety of reagents that can in principle be employed in a conversion reaction, or otherwise to take specific measures for achieving a desired conversion, for example to use protection group techniques. However, finding out suitable reaction variants and reaction conditions in such cases does not cause any problems for one skilled in the art.
The structural elements present in the residues attached at the 1-position of the pyrazole ring in the compound of formulae I, Ib and Ic and in the COR8′ group present in the 3-position and/or in the 5-position of the pyrazole ring can be introduced into the starting pyrazole derivative obtainable as outlined above by consecutive reaction steps using synthesis methodologies like those outlined below using procedures which per se are well known to one skilled in the art.
The residues R8′ that can be introduced in formula 2, for example, by condensing a corresponding carboxylic acid of the formula 2 with a compound of the formula HR8′, i.e. with an amine of the formula HN(R1′)R2′—V-G-M to give a compound of the formula 3. the compound of the formula 3 thus obtained can already contain the desired final groups, i.e. the groups R8′ and R8
Thus, the residues R8′ and the residues R1′ and R2′-V-G-M contained therein can have the denotations of R1 and R2—V-G-M, respectively, given above or in addition in the residues R1′ and R2′—V-G-M functional groups can also be present in the form of groups that can subsequently be transformed into the final groups R1 and R2—V-G-M, i.e. functional groups can be present in the form of precursor groups or of derivatives, for example in protected form. In the course of the preparation of the compound of formulae I, Ib and Ic, it can generally be advantageous or necessary to introduce functional groups which reduce or prevent undesired reactions or side reactions in the respective synthesis step, in the form of precursor groups which are later converted into the desired functional groups, or to temporarily block functional groups by a protective group strategy suited to the synthesis problem. Such strategies are well known to those skilled in the art (see, for example, Greene and Wuts, Protective Groups in Organic Synthesis, Wiley, 1991, or P. Kocienski, Protecting Groups, Thieme 1994). Examples of precursor groups are cyano and nitro. The cyano group can in a later step be transformed into carboxylic acid derivatives or by reduction into aminomethyl groups, or the nitro groups may be transformed by reduction like catalytic hydrogenation into amino groups. Protective groups can also have the meaning of a solid phase, and cleavage from the solid phase stands for the removal of the protective group. The use of such techniques is known to those skilled in the art (Burgess K (Ed.) Solid Phase Organic Synthesis, New York, Wiley, 2000). For example, a phenolic hydroxy group can be attached to a trityl-polystyrene resin, which serves as a protecting group, and the molecule is cleaved from this resin by treatment with TFA at a later stage of the synthesis.
The residue R87 in the compound of formulae 2 and 3 can denote the group -Q-R0 as defined above which finally is to be present in the desired target molecule of the formulae I, Ib and Ic, or it can denote a group which can subsequently be transformed into the group -Q-R0, for example a precursor group or a derivative of the group -Q-R0 in which functional groups are present in protected form, or R87 can denote a hydrogen atom or a protective group for the nitrogen atom of the pyrazole ring. Similarly, the residues R1
The residues R86 in the compound of formula 2 which can be identical or different, can be, for example, hydroxy or (C1-C4)-alkoxy, i.e., the groups COR86 present in the compound of formula 2 can be, for example, the free carboxylic acids or esters thereof like alkyl esters as can be the groups COR8′ in the compound of formulae I, Ib and Ic. The groups COR86 can also be any other activated derivative of a carboxylic acid which allows amide formation, ester formation or thioester formation with a compound of the formula HR8′. The group COR86 can be, for example, an acid chloride, an activated ester like a substituted phenyl ester or an N-hydroxysuccinimide or a hydroxybenzotriazole ester, an azolide like an imidazolide, an azide or a mixed anhydride, for example a mixed anhydride with a carbonic acid ester or with a sulfonic acid, which derivatives can all be prepared from the carboxylic acid by standard procedures and can be reacted with an amine, an alcohol or a mercaptan of the formula HR8′ under standard conditions. A carboxylic acid group COOH representing COR86 in a compound of the formula 2 can be obtained, for example, from an ester group introduced into the pyrazole system during a pyrazole synthesis by standard hydrolysis procedures. It can also be obtained, for example, by hydrolysis of a nitrile group introduced into the pyrazole system during a pyrazole synthesis.
A compound of formulae I, Ib and Ic in which a group COR8′ is an ester group can also be prepared from a compound of formula 2 in which COR86 is a carboxylic acid group by common esterification reactions like, for example, reacting the acid with an alcohol under acid catalysis, or alkylation of a salt of the carboxylic acid with an electrophile like an alkyl halogenide, or by transesterification from another ester. A compound of formulae I, Ib and Ic in which a group COR8′ is an amide group can be prepared from amines and a compound of formula 2 in which COR86 is a carboxylic acid group or an ester thereof by common amination reactions. Especially for the preparation of amides the compound of formula 2 in which COR86 is a carboxylic acid group can be condensed under standard conditions with a compound of formula HR8′ which are amines by means of common coupling reagents used in peptide synthesis. Such coupling reagents are, for example, carbodiimides like dicyclohexylcarbodiimide (DCC) or diisopropylcarbodiimide, carbonyldiazoles like carbonyldiimidazole (CDI) and similar reagents, propylphosphonic anhydride, O-((cyano-(ethoxycarbonyl)-methylene)amino)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TOTU), diethylphosphoryl cyanide (DEPC) or bis-(2-oxo-3-oxazolidinyl)-phosphoryl chloride (BOP-Cl) and many others.
If the residue -Q-R0 present in an pyrazole of the formulae I, Ib and Ic or the residue R87 present in an pyrazole of the formula 2, or a residue in which functional groups within the residue -Q-R0 or R87 are present in protected form or in the form of a precursor group, have not already been introduced during a preceding step, for example during a synthesis of the pyrazole nucleus, these residues can, for example, be introduced into the 1-position of the pyrazole system by conventional literature procedures well known to one skilled in the art for N-alkylation, reductive amination, N-arylation, N-acylation or N-sulfonylation of ring nitrogen atoms of heterocycles. The starting pyrazole derivative that is to be employed in such a reaction carries a hydrogen atom in the 1-position. N-Alkylation of a ring nitrogen atom can, for example, be performed under standard conditions, preferably in the presence of a base like K2CO3, Cs2CO3, NaH or KOtBu, using an alkylating compound of the formula LG-Q-R0 or of the formula R87-LG, wherein the atom in the group Q or in the group R87 bonded to the group LG in this case is an aliphatic carbon atom of an alkyl moiety and LG is a leaving group, for example halogen like chlorine, bromine or iodine, or a sulfonyloxy group like tosyloxy, mesyloxy or trifluormethylsulfonyloxy. LG may, for example, also be a hydroxy group which, in order to achieve the alkylation reaction, is activated in a well-known Mitsunobu reaction by a conventional activating agent. The regioselectivity of the N-alkylation can be controlled by the choice of the base, solvent and reaction conditions. Nevertheless mixtures of positional isomers, can be separated by modern separation techniques like, for example, flash chromatography, crystallisation or preparative HPLC.
For the preparation of compounds in which A is a direct linkage and an aromatic group is directly bonded to the 1-position of the pyrazole system, conventional arylation procedures can be used. For example aryl fluorides like alkyl fluorobenzoates or 4-fluorophenyl nitriles can be employed as arylating agents. Such processes are described, for example, by K. Cooper et al., J. Med. Chem. (1992), 35, 3115; M. Artico et al., Eur. J. Med. Chem. Chim. Ther. (1992) 27, 219; X.-J. Wang et al., Tetrahedron Letters (2000) 41, 5321; M. L. Cerrada et al., Synth. Commun. (1993) 23, 1947. Alternatively a wide variety of substituted aryl iodides, aryl bromides or aryl triflates can serve as arylating agents at the 1-position of the heterocyclic nitrogen in a copper salt or palladium mediated reaction according for example to P. Cozzi et al. Farmaco (1987) 42, 205; P. Unangst, D. Connor, R. Stabler, R. Weikert, J. Heterocycl. Chem. (1987) 24, 811; G. Tokmakov, I. Grandberg, Tetrahedron (1995) 51, 2091; D. Old, M. Harris, S. Buchwald, Org. Lett. (2000) 2, 1403, G. Mann, J. Hartwig, M. Driver, C. Fernandez-Rivas, J. Am. Chem. Soc. (1998) 120, 827; J. Hartwig, M. Kawatsura, S. Hauk, K. Shaughnessy, L. J. Org. Chem. (1999) 64, 5575; S. Buchwald et al., J. Am. Chem. Soc. (2001) 123, 7727. Moreover such arylations can also be accomplished by reaction of a wide range of substituted aryl boronic acids as demonstrated for example by P. Lam et al., Tetrahedron Lett. (1998) 39, 2941; V. Collot et al., Tetrahedron Lett. (2000) 41, 9053; P. Lam et al., Tetrahedron Lett. (2001) 42, 3415;
Preferred methods include, but are not limited to those described in the examples.
The compounds of the present invention are serine protease inhibitors, which inhibit the activity of the blood coagulation enzyme factors Xa and/or factor VIIa. In particular, they are highly active inhibitors of factor Xa. They are specific serine protease inhibitors inasmuch as they do not substantially inhibit the activity of other proteases whose inhibition is not desired. The activity of the compound of formulae I, Ib and Ic can be determined, for example, in the assays described below or in other assays known to those skilled in the art. With respect to factor Xa inhibition, a preferred embodiment of the invention comprises compounds which have a Ki<1 mM for factor Xa inhibition as determined in the assay described below, with or without concomitant factor VIIa inhibition, and which preferably do not substantially inhibit the activity of other proteases involved in coagulation and fibrinolysis whose inhibition is not desired (using the same concentration of the inhibitor). the compounds of the invention inhibit factor Xa catalytic activity either directly, within the prothrombinase complex or as a soluble subunit, or indirectly, by inhibiting the assembly of factor Xa into the prothrombinase complex.
As inhibitors of factor Xa and/or factor VIIa the compound of formulae I, Ib and Ic and their physiologically tolerable salts and their prodrugs are generally suitable for the therapy and prophylaxis of conditions in which the activity of factor Xa and/or factor VIIa plays a role or has an undesired extent, or which can favorably be influenced by inhibiting factor Xa and/or factor VIIa or decreasing their activities, or for the prevention, alleviation or cure of which an inhibition of factor Xa and/or factor VIIa or a decrease in their activity is desired by the physician. As inhibition of factor Xa and/or factor VIIa influences blood coagulation and fibrinolysis, the compound of formulae I, Ib and Ic and their physiologically tolerable salts and their prodrugs are generally suitable for reducing blood clotting, or for the therapy and prophylaxis of conditions in which the activity of the blood coagulation system plays a role or has an undesired extent, or which can favorably be influenced by reducing blood clotting, or for the prevention, alleviation or cure of which a decreased activity of the blood coagulation system is desired by the physician. A specific subject of the present invention thus are the reduction or inhibition of unwanted blood clotting, in particular in an individual, by administering an effective amount of a compound I or a physiologically tolerable salt or a prodrug thereof, as well as pharmaceutical preparations therefor.
The present invention also is directed to the compound of formulae I, Ib and Ic and/or their physiologically tolerable salts and/or their prodrugs for use as pharmaceuticals (or medicaments), to the use of the compound of formulae I, Ib and Ic and/or their physiologically tolerable salts and/or their prodrugs for the production of pharmaceuticals for inhibition of factor Xa and/or factor VIIa or for influencing blood coagulation, inflammatory response or fibrinolysis or for the therapy or prophylaxis of the diseases mentioned above or below, for example for the production of pharmaceuticals for the therapy and prophylaxis of cardiovascular disorders, thromboembolic diseases or restenoses. The invention also is directed to the use of the compound of formulae I, Ib and Ic and/or their physiologically tolerable salts and/or their prodrugs for the inhibition of factor Xa and/or factor VIIa or for influencing blood coagulation or fibrinolysis or for the therapy or prophylaxis of the diseases mentioned above or below, for example for use in the therapy and prophylaxis of cardiovascular disorders, thromboembolic diseases or restenoses, and to methods of treatment aiming at such purposes including methods for said therapies and prophylaxis. The present invention also is directed to pharmaceutical preparations (or pharmaceutical compositions) which contain an effective amount of at least one compound of the formulae I, Ib and Ic and/or its physiologically tolerable salts and/or its prodrugs in addition to a customary pharmaceutically acceptable carrier, i.e. one or more pharmaceutically acceptable carrier substances or excipients and/or auxiliary substances or additives.
The invention also is directed to the treatment of disease states such as abnormal thrombus formation, acute myocardial infarction, unstable angina, thromboembolism, acute vessel closure associated with thrombolytic therapy or percutaneous transluminal coronary angioplasty (PTCA), transient ischemic attacks, stroke, intermittent claudication or bypass grafting of the coronary or peripheral arteries, vessel luminal narrowing, restenosis post coronary or venous angioplasty, maintenance of vascular access patency in long-term hemodialysis patients, pathologic thrombus formation occurring in the veins of the lower extremities following abdominal, knee or hip surgery, pathologic thrombus formation occurring in the veins of the lower extremities following abdominal, knee and hip surgery, a risk of pulmonary thromboembolism, or disseminated systemic intravascular coagulatopathy occurring in vascular systems during septic shock, certain viral infections or cancer. The compounds of the present invention can also be used to reduce an inflammatory response. Examples of specific disorders for the treatment or prophylaxis of which the compound of formulae I, Ib and Ic can be used are coronary heart disease, myocardial infarction, angina pectoris, vascular restenosis, for example restenosis following angioplasty like PTCA, adult respiratory distress syndrome, multi-organ failure and disseminated intravascular clotting disorder. Examples of related complications associated with surgery are thromboses like deep vein and proximal vein thrombosis, which can occur following surgery.
The compound of formulae I, Ib and Ic and their physiologically tolerable salts and their prodrugs can be administered to animals, preferably to mammals, and in particular to humans as pharmaceuticals for therapy or prophylaxis. They can be administered on their own, or in mixtures with one another or in the form of pharmaceutical preparations, which permit enteral or parenteral administration.
The pharmaceuticals can be administered orally, for example in the form of pills, tablets, lacquered tablets, coated tablets, granules, hard and soft gelatin capsules, solutions, syrups, emulsions, suspensions or aerosol mixtures. Administration, however, can also be carried out rectally, for example in the form of suppositories, or parenterally, for example intravenously, intramuscularly or subcutaneously, in the form of injection solutions or infusion solutions, microcapsules, implants or rods, or percutaneously or topically, for example in the form of ointments, solutions or tinctures, or in other ways, for example in the form of aerosols or nasal sprays.
The pharmaceutical preparations according to the invention are prepared in a manner known per se and familiar to one skilled in the art, pharmaceutically acceptable inert inorganic and/or organic carriers being used in addition to the compound(s) of the formulae I, Ib and Ic and/or its (their) physiologically tolerable salts and/or its (their) prodrugs. For the production of pills, tablets, coated tablets and hard gelatin capsules it is possible to use, for example, lactose, cornstarch or derivatives thereof, talc, stearic acid or its salts, etc. Carriers for soft gelatin capsules and suppositories are, for example, fats, waxes, semisolid and liquid polyols, natural or hardened oils, etc. Suitable carriers for the production of solutions, for example injection solutions, or of emulsions or syrups are, for example, water, saline, alcohols, glycerol, polyols, sucrose, invert sugar, glucose, vegetable oils, etc. Suitable carriers for microcapsules, implants or rods are, for example, copolymers of glycolic acid and lactic acid. The pharmaceutical preparations normally contain about 0.5% to 90% by weight of the compound of formulae I, Ib and Ic and/or their physiologically tolerable salts and/or their prodrugs. The amount of the active ingredient of the formulae I, Ib and Ic and/or its physiologically tolerable salts and/or its prodrugs in the pharmaceutical preparations normally is from about 0.5 mg to about 1000 mg, preferably from about 1 mg to about 500 mg.
In addition to the active ingredients of the formulae I, Ib and Ic and/or their physiologically acceptable salts and/or prodrugs and to carrier substances, the pharmaceutical preparations can contain additives such as, for example, fillers, disintegrants, binders, lubricants, wetting agents, stabilizers, emulsifiers, preservatives, sweeteners, colorants, flavorings, aromatizers, thickeners, diluents, buffer substances, solvents, solubilizers, agents for achieving a depot effect, salts for altering the osmotic pressure, coating agents or antioxidants. They can also contain two or more compounds of formulae I, Ib and Ic, and/or their physiologically tolerable salts and/or their prodrugs. In case a pharmaceutical preparation contains two or more a compound of formulae I, Ib and Ic, the selection of the individual compounds can aim at a specific overall pharmacological profile of the pharmaceutical preparation. For example, a highly potent compound with a shorter duration of action may be combined with a long-acting compound of lower potency. The flexibility permitted with respect to the choice of substituents in the compound of formulae I, Ib and Ic allows a great deal of control over the biological and physico-chemical properties of the compounds and thus allows the selection of such desired compounds. Furthermore, in addition to at least one compound of formulae I, Ib and Ic and/or its physiologically tolerable salt and/or its prodrug, the pharmaceutical preparations can also contain one or more other therapeutically or prophylactically active ingredients.
When using the compound of formulae I, Ib and Ic the dose can vary within wide limits and, as is customary and is known to the physician, is to be suited to the individual conditions in each individual case. It depends, for example, on the specific compound employed, on the nature and severity of the disease to be treated, on the mode and the schedule of administration, or on whether an acute or chronic condition is treated or whether prophylaxis is carried out. An appropriate dosage can be established using clinical approaches well known in the medical art. In general, the daily dose for achieving the desired results in an adult weighing about 75 kg is from 0.01 mg/kg to 100 mg/kg, preferably from 0.1 mg/kg to 50 mg/kg, in particular from 0.1 mg/kg to 10 mg/kg, (in each case in mg per kg of body weight). The daily dose can be divided, in particular in the case of the administration of relatively large amounts, into several, for example 2, 3 or 4, part administrations. As usual, depending on individual behavior it may be necessary to deviate upwards or downwards from the daily dose indicated.
A compound of the formulae I, Ib and Ic can also advantageously be used as an anticoagulant outside an individual. For example, an effective amount of a compound of the invention can be contacted with a freshly drawn blood sample to prevent coagulation of the blood sample. Further, a compound of the formulae I, Ib and Ic or its salts can be used for diagnostic purposes, for example in in vitro diagnoses, and as an auxiliary in biochemical investigations. For example, a compound of the formulae I, Ib and Ic can be used in an assay to identify the presence of factor Xa and/or factor VIIa or to isolate factor Xa and/or factor VIIa in a substantially purified form. A compound of the invention can be labeled with, for example, a radioisotope, and the labeled compound bound to factor Xa and/or factor VIIa is then detected using a routine method useful for detecting the particular label. Thus, a compound of the formulae I, Ib and Ic or a salt thereof can be used as a probe to detect the location or amount of factor Xa and/or factor VIIa activity in vivo, in vitro or ex vivo.
Furthermore, the compound of formulae I, Ib and Ic can be used as synthesis intermediates for the preparation of other compounds, in particular of other pharmaceutical active ingredients, which are obtainable from the compound of formulae I, Ib and Ic, for example by introduction of substituents or modification of functional groups.
The general synthetic sequences for preparing the compounds useful according to the present invention are outlined in the examples given below. Both an explanation of, and the actual procedure for, the various aspects of the present invention are described where appropriate. The following examples are intended to be merely illustrative of the present invention, and not limiting thereof in either scope or spirit. Those with skill in the art will readily understand that known variations of the conditions and processes described in the examples can be used to synthesize the compounds of the present invention.
It is understood that changes that do not substantially affect the activity of the various embodiments of this invention are included within the invention disclosed herein. Thus, the following examples are intended to illustrate but not limit the present invention.
When in the final step of the synthesis of a compound an acid such as trifluoroacetic acid or acetic acid was used, for example when trifluoroacetic acid was employed to remove a t-Bu group or when a compound was purified by chromatography using an eluent which contained such an acid, in some cases, depending on the work-up procedure, for example the details of a freeze-drying process, the compound was obtained partially or completely in the form of a salt of the acid used, for example in the form of the acetic acid salt or trifluoroacetic acid salt or hydrochloric acid salt.
To a solution of 5.0 g Piperidin-4-yl-carbamic acid tert-butyl ester in 15 ml methanol 7.34 ml acetone, 3.14 g Na(CN)BH3 and 0.3 ml acetic acid were added. After stirring for 16 h at RT the solvent was removed under reduced pressure and the residue was partitioned between 30 ml of water and 30 ml of ethyl acetate. The organic layer was washed with saturated Na2CO3 solution, water and then dried over Na2SO4. Following filtration, the solvent was removed under reduced pressure to yields a white solid. Yield: 4.8 g MS (ES+): m/e=243.
To 4.8 g (1-Isopropyl-piperidin-4-yl)-carbamic acid tert-butyl ester in 15 ml methanol, 20 ml methanolic hydrochloric acid (8M) were added and the mixture was stirred for 16 h. Removal of the solvent under reduced pressure yielded a white solid, which was coevaporated twice with 20 ml toluene. The product was obtained as its hydrochloride.
Yield: 5.42 g MS (ES+): m/e=143.
To a solution of 2.0 g 5-Thiophen-2-yl-2H-pyrazole-3-carboxylic acid ethyl ester in 5 ml DMF, 360 mg NaH (60% in mineral oil) and subsequently 2.8 g 3-Bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole [prepared by adopting a procedure described by Ewing, William R.; Becker, Michael R.; Choi-Sledeski, Yong Mi; Pauls, Heinz W.; He, Wei; Condon, Stephen M.; Davis, Roderick S.; Hanney, Barbara A.; Spada, Alfred P.; Burns, Christopher J.; Jiang, John Z.; Li, Aiwen; Myers, Michael R.; Lau, Wan F.; Poli, Gregory B; PCT Int. Appl. (2001) 460 pp. WO 0107436 A2] were added and the mixture was stirred at 80° C. for 1 h. After the addition of 5 ml water the mixture was filtered through a Chem Elut® cartridge by elution with ethyl acetate and then concentrated under reduced pressure. The residue was directly subjected to the subsequent saponification reaction without further purification. Yield: 4 g.
To a solution of 4 g 2-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-thiophen-2-yl-2H-pyrazole-3-carboxylic acid ethyl ester in 20 ml THF, 10 ml water and 500 mg lithium hydroxide monohydrate were added. After stirring for 2 h at 60° C. the reaction was cooled to RT. The mixture was acidified with half concentrated hydrochloric acid to pH 3 and the precipitate collected by filtration and washed with 10 ml water. The product was obtained as a white solid which was dried under reduced pressure. Yield: 3.8 g.
To a solution of 200 mg 2-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-thiophen-2-yl-2H-pyrazole-3-carboxylic acid, 0.3 ml N-NEM in 2 ml DCM, 168 mg TOTU were added and the mixture was stirred for 30 min at RT. Then 136 mg 1-Isopropyl-piperidin-4-ylamine hydrochloride were added and the reaction was further stirred for 2 h. After the addition of 2 ml sat. NaHCO3 the mixture was filtered through a Chem Elut® cartridge by elution with ethyl acetate and then concentrated under reduced pressure. The residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. The product was obtained as its trifluoroacetate salt. Yield: 120 mg MS (ES+): m/e=516, chloro pattern.
This compound was isolated as a by-product in example 1.
MS (ES+): m/e=516, chloro pattern.
The title compound was prepared analogously to example 1 with the difference that 5-Methyl-2H-pyrazole-3-carboxylic acid methyl ester was used instead of 5-Thiophen-2-yl-2H-pyrazole-3-carboxylic acid ethyl ester. MS (ESI+): m/e=448, chloro pattern.
The title compound was prepared analogously to example 1 with the difference that 2-(6-Chloro-benzothiazol-2-yl)-5-methyl-2H-pyrazole-3-carboxylic acid was used instead of of 5-Thiophen-2-yl-2H-pyrazole-3-carboxylic acid ethyl ester. MS (ESI+): m/e=418, chloro pattern.
The title compound was prepared analogously to example 1 with the difference that 5-(5-Chloro-thiophen-2-yl)-2H-pyrazole-3-carboxylic acid methyl ester was used instead of of 5-Thiophen-2-yl-2H-pyrazole-3-carboxylic acid ethyl ester. MS (ESI+): m/e=550, chloro pattern.
This compound was isolated as a by-product in example 5.
MS (ES+): m/e=550, chloro pattern.
The title compound was prepared analogously to example 1 with the difference that 4-(2,4-Dichloro-phenyl)-2H-pyrazole-3-carboxylic acid methyl ester was used instead of 5-Thiophen-2-yl-2H-pyrazole-3-carboxylic acid ethyl ester. MS (ESI+): m/e=578, chloro pattern.
The title compound was prepared analogously to example 1 with the difference that 5-Propyl-2H-pyrazole-3-carboxylic acid methyl ester was used instead of 5-Thiophen-2-yl-2H-pyrazole-3-carboxylic acid ethyl ester. MS (ESI+): m/e=476, chloro pattern.
The title compound was prepared analogously to example 1 with the difference that 5-tert-Butyl-2H-pyrazole-3-carboxylic acid methyl ester was used instead of 5-Thiophen-2-yl-2H-pyrazole-3-carboxylic acid ethyl ester. MS (ESI+): m/e=490, chloro pattern.
1.0 g (5.5 mmol) of 5-propyl-2H-pyrazole-3-carboxylic acid ethyl ester was dissolved in 15 ml of dichloromethane and 1.23 g (5.5 mmol) of N-iodosuccinimide was added. The resulting solution was stirred at room temperature for 16 h. The solution was washed with aqueous sodium thiosulfate solution. The organic phase was dried with sodium sulfate and filtered. The resulting solution was passed through a short silica gel column, washing with dichloromethane. The solvent was removed under reduced pressure.
Yield: 1.5 g MS (LCMS-ES+): m/e=309.
1.5 g (4.9 mmol) of 4-Iodo-5-propyl-2H-pyrazole-3-carboxylic acid ethyl ester, 0.87 g (9.7 mmol) of copper cyanide and 404 mg (2.4 mmol) of tetraethylammonium cyanide were dissolved in 10 ml of DMF and 20 ml of tetrahydrofuran and the solution was degassed with argon. 223 mg (0.2 mmol) of Tris(dibenzylideneacetone)dipalladium (0) and 404 mg (0.7 mmol) of 1,1′-bis-(diphenylphosphino) ferrocene were added at RT. The reaction was stirred at 120° C. for 5 h. The solvent was removed under reduced pressure. The residue was dissolved in ethyl acetate and this solution was washed with saturated aqueous sodium bicarbonate. The organic phase was dried with sodium sulfate, filtered and the solvent was removed under reduced pressure. The product was purified by silica gel chromatography eluting with n-heptane:ethyl acetate/1:1.
Yield: 110 mg MS (LCMS-ES+): m/e=208.
112 mg (0.5 mmol) of 4-cyano-5-propyl-2H-pyrazole-3-carboxylic acid ethyl ester was dissolved in 2 ml of DMF and 23.8 mg (0.6 mmol) of sodium hydride (60% in mineral oil) were added at RT. After stirring for 20 min at room temperature the solution was cooled to −70° C. and 166 mg (0.6 mmol) of 3-bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole were added. The reaction was stirred at room temperature for 3 h. The reaction solution was treated with 1 ml of 2N aqueous NaOH for 16 h at room temperature. The product was purified by preparative RP-HPLC eluting with a gradient of 0-100% acetonitrile in water (+0.01% trifluoroacetic acid). After lyophilization the product was obtained as a white solid.
Yield 55.3 mg. MS (LCMS-ES+): m/e=377, chloro pattern.
To a solution of 55 mg 2-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-4-cyano-5-propyl-2H-pyrazole-3-carboxylic acid, 0.1 ml N-NEM in 2 ml DMF, 48 mg TOTU were added and the mixture was stirred for 30 min at RT. Then 31 mg of 1-Isopropyl-piperidin-4-ylamine hydrochloride were added and the reaction was further stirred for 2 h. After addition of 2 ml sat. NaHCO3 the mixture was filtered through a Chem Elut® cartridge by elution with ethyl acetate and then concentrated under reduced pressure. After removal of the solvent under reduced pressure the residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. The product was obtained as its trifluoroacetate salt.
Yield: 18 mg MS (ES+): m/e=501, chloro pattern.
This compound was isolated as a by-product in example 10.
MS (ES+): m/e=501, chloro pattern.
1.0 g (4.5 mmol) of 5-thiophen-2-yl-2H-pyrazole-3-carboxylic acid ethyl ester was dissolved in 15 ml of dichloromethane and 1.01 g (4.5 mmol) of N-iodosuccinimide was added. The resulting solution was stirred at room temperature for 16 h. The solution was washed with aqueous sodium thiosulfate solution. The organic phase was dried with sodium sulfate and filtered. The resulting solution was passed through a short silica gel column, washing with dichloromethane. The solvent was removed under reduced pressure.
Yield: 1.57 g MS (LCMS-ES+): m/e=349.
1.57 g (4.5 mmol) of 4-Iodo-5-thiophen-2-yl-2H-pyrazole-3-carboxylic acid ethyl ester, 0.81 g (9.0 mmol) of copper cyanide and 352 mg (2.3 mmol) of tetraethylammonium cyanide were dissolved in 10 ml of DMF and 20 ml of tetrahydrofuran and the solution was degassed with argon. 223 mg (0.2 mmol) of Tris(dibenzylideneacetone)dipalladium (0) and 374 mg (0.7 mmol) of 1,1′-bis-(diphenylphosphino) ferrocene were added at RT. The reaction was stirred at 120° C. for 5 h. The solvent was removed under reduced pressure. The residue was dissolved in ethyl acetate and this solution was washed with saturated aqueous sodium bicarbonate. The organic phase was dried with sodium sulfate, filtered and the solvent was removed under reduced pressure. The product was purified by silica gel chromatography eluting with n-heptane:ethyl acetate/1:1.
Yield: 287 mg MS (LCMS-ES+): m/e=248.
287 mg (1.2 mmol) of 4-Cyano-5-thiophen-2-yl-2H-pyrazole-3-carboxylic acid ethyl ester was dissolved in 2 ml of DMF and 51.1 mg (1.3 mmol) of sodium hydride (60% in mineral oil) were added at RT. After stirring for 20 min at room temperature the solution was cooled to −70° C. and 355 mg (1.3 mmol) of 3-bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole were added. The reaction was stirred at room temperature for 3 h. The reaction solution was treated with 1 ml of 2N aqueous NaOH for 16 h at room temperature. The product was purified by preparative RP-HPLC eluting with a gradient of 0-100% acetonitrile in water (+0.01% trifluoroacetic acid). After lyophilization the product was obtained as a white solid.
Yield 122 mg. MS (LCMS-ES+): m/e=417.
To a solution of 31 mg 2-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-4-cyano-5-thiophen-2-yl-2H-pyrazole-3-carboxylic acid, 0.1 ml N-NEM in 1 ml DMF, 24 mg TOTU were added and the mixture was stirred for 30 min at RT. Then 16 mg 1-Isopropyl-piperidin-4-ylamine hydrochloride were added and the reaction was further stirred for 2 h. After addition of 2 ml sat. NaHCO3 the mixture was filtered through a Chem Elut® cartridge by elution with ethyl acetate and then concentrated under reduced pressure. After removal of the solvent under reduced pressure the residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. The product was obtained as its trifluoroacetate salt.
Yield: 18 mg MS (ES+): m/e=541, chloro pattern.
This compound was isolated as a by-product in example 12.
MS (ES+): m/e=541, chloro pattern.
The title compound was prepared analogously to example 1 with the difference that 5-Bromomethyl-2-(5-chloro-thiophen-2-yl)-thiazole [prepared by adopting a procedure described by Ewing, William R. et al.; PCT Int. Appl. (2001) 460 pp. WO 0107436 A2] was used instead of 3-Bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole. MS (ES+): m/e=532, chloro pattern.
The title compound was prepared analogously to example 1 with the difference that 2-Bromomethyl-6-chloro-benzo[b]thiophene [prepared by adopting a procedure described by Ewing, William R. et al.; PCT Int. Appl. (1999) 300 pp. WO 9937304 A1; and Ewing, William R. et al. PCT Int. Appl. (2001) 460 pp. WO 0107436 A2] was used instead of 3-Bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole. MS (ES+): m/e=499, chloro pattern.
A suspension of 5 g (23.3 mmol) Piperidin-4-ylmethyl-carbamic acid tBu ester, 3.85 g (25.7 mmol) 4-Chloropyridine hydrochloride in 15 ml n-BuOH/H2O/NEt3 1:1:1 was boiled under reflux for 3 days. After removal of the solvent under reduced pressure the residue was purified by chromatography on silica with DCM/MeOH 100:1->50:1->10:1->5:1 to yield a white solid.
Yield: 4.3 g.
To a solution of 4.58 g (3,4,5,6-Tetrahydro-2H-[1,4′]bipyridinyl-4-ylmethyl)-carbamic acid tBu ester in 12 ml DCM, 12 ml of TFA were added at RT. After stirring for 30 min the solution was diluted with 20 ml of toluene and then evaporated under reduced pressure. The residue was codestilled twice with toluene and was used in the subsequent reactions without further purification. The product was obtained as its trifluoroacetate salt. Yield: 3.3 g.
To a solution of 50 mg 2-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-thiophen-2-yl-2H-pyrazole-3-carboxylic acid, 0.3 ml N-NEM in 1 ml DCM, 59 mg TOTU were added and the mixture was stirred for 30 min at RT. Then 36 mg of C-(3,4,5,6-Tetrahydro-2H-[1,4′]bipyridinyl-4-yl)-methylamine trifluoroacetate were added and the reaction was further stirred for 2 h. After the addition of 2 ml sat. NaHCO3 the mixture was filtered through a Chem Elut® cartridge by elution with ethyl acetate and then concentrated under reduced pressure. The residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. The product was obtained as its trifluoroacetate salt.
Yield: 23 mg MS (ES+): m/e=565, chloro pattern.
To a solution of 1.0 g Piperidin-4-ylmethyl-carbamic acid tert-butyl ester in 20 ml acetonitrile 2.6 ml acetone and 586 mg Na(CN)BH3 were added. After stirring for 16 h at RT the solvent was removed under reduced pressure and the residue was partitioned between 30 ml of water and 30 ml of ethylacetate. The organic layer was washed with saturated Na2CO3 solution, water and then was dried over Na2SO4. Removal of the solvent under reduced pressure yielded a white solid. Yield: 802 mg.
To a solution of 802 mg (1-Isopropyl-piperidin-4-ylmethyl)-carbamic acid tert-butyl ester in 5 ml DCM 4 ml of TFA were added at RT. After stirring for 20 h the solution was diluted with 20 ml of toluene and the solvents were evaporated under reduced pressure. The residue was codestilled twice with toluene and used in the subsequent reaction without further purification. The product was obtained as its trifluoroacetate salt. Yield: 1.7 g
To a solution of 50 mg 2-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-thiophen-2-yl-2H-pyrazole-3-carboxylic acid, 0.3 ml N-NEM in 1 ml DCM, 59 mg TOTU were added and the mixture was stirred for 30 min at RT. Then 30 mg C-(1-Isopropyl-piperidin-4-yl)-methylamine trifluoroacetate were added and the reaction was stirred for a further 2 h. After the addition of 2 ml sat. NaHCO3 the mixture was filtered through a Chem Elut® cartridge by elution with ethyl acetate and then concentrated under reduced pressure. After removal of the solvent under reduced pressure the residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. The product was obtained as its trifluoroacetate salt. Yield: 14 mg MS (ES+): m/e=530, chloro pattern.
A solution of 3 g Piperidin-4-yl-carbamic acid tert-butyl ester and 2.5 g 4-Chloropyridine in 9 ml n-butanol/water/NEt3 1:1:1 was heated at 100° C. for 48 h. Then the solution was cooled to RT diluted with DCM and washed with NaHCO3 solution and water. The organic layer was dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. Chromatographic purification of the residue on silica with DCM as eluent gave after evaporation of the fractions containing the product a white foam. Yield 1.7 g.
To a solution of 4 g (3,4,5,6-Tetrahydro-2H-[1,4′]bipyridinyl-4-yl)-carbamic acid tert-butyl ester in 4 ml DCM, 12 ml TFA were added at RT. After stirring for 20 h the solution was diluted with 20 ml of toluene and the solvents were evaporated under reduced pressure. The residue was codestilled twice with toluene and then used in the subsequent reaction without further purification. The product was obtained as its trifluoroacetate salt. Yield: 2.7 g.
To a solution of 50 mg 2-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-thiophen-2-yl-2H-pyrazole-3-carboxylic acid, 0.3 ml N-NEM in 1 ml DCM, 59 mg TOTU were added and the mixture was stirred for 30 min at RT. Then 33 mg 3,4,5,6-Tetrahydro-2H-[1,4′]bipyridinyl-4-ylamine trifluoroacetate were added and the reaction was further stirred for 2 h. After the addition of 2 ml sat. NaHCO3 the mixture was filtered through a Chem Elut® cartridge by elution with ethyl acetate and then concentrated under reduced pressure. The residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. The product was obtained as its trifluoroacetate salt. Yield: 25 mg MS (ES+): m/e=551, chloro pattern.
The title compound was prepared analogously to example 12 with the difference that 1-Bromomethyl-4-chloro-benzene was used instead of 3-Bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole in the alkylation step. MS (ESI+): m/e=468, chloro pattern.
To 12 g of 1H-Pyrazole-3,5-dicarboxylic acid 100 ml HCl in methanol (8M) were added at RT and stirred for 48 h. Then the solvents were removed under reduced pressure and the residue codestilled with toluene (2×50 ml). Yield: 14 g.
To a solution of 1 g 1H-Pyrazole-3,5-dicarboxylic acid diethyl ester in 20 ml of DMF and 188 mg of sodium hydride (60% in mineral oil) were added at RT. After stirring for 20 min at room temperature 1.32 g of 3-bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole were added. The reaction was stirred at room temperature for 3 h. Then 100 ml water were added and the precipitating product was collected by filtration. Yield: 1.7 g.
To a solution of 1.7 g 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-1H-pyrazole-3,5-dicarboxylic acid dimethyl ester in 10 ml water/THF 1:1, 4 ml of a 1M aqueous NaOH were added at RT and the mixture was stirred for 3 h with LCMS reaction control. Then the reaction mixture was acidified to pH 3 with half concentrated HCl and extracted with DCM (3×50 ml). The organic phase was dried over MgSO4, filtered and the solvent removed under reduced pressure. The residue was subjected to the next reaction step without further purification.
To 1 g 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-1H-pyrazole-3,5-dicarboxylic acid 3-methyl ester in 10 ml DCM and 1.4 ml NEt3, 667 mg BOP-Cl were added at RT and the mixture was stirred for 30 min. After addition of 563 mg 1-Isopropyl-piperidin-4-ylamine hydrochloride the mixture was stirred for 16 h. After removal of the solvent under reduced pressure the residue was purified by silica gel chromatography eluting with DCM/MeOH/AcOH/H2O 20:10:1:1 to yield a white solid. Yield: 800 mg MS (ES+): m/e=492, chloro pattern.
To a solution of 800 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid methyl ester in 5 ml water/THF 1:1, 3 ml of a 1M aqueous NaOH were added at RT and the mixture was heated for 10 h at 60° C. Then the reaction mixture was acidified to pH 3 with half concentrated HCl and extracted with DCM (3×50 ml). The organic phase was dried over MgSO4, filtered and the solvent removed under reduced pressure. The residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. The product was obtained as its trifluoroacetate salt.
Yield: 503 mg MS (ES+): m/e=478, chloro pattern.
This compound was isolated as a by-product in example 21.
MS (ES+): m/e=478, chloro pattern.
The title compound was prepared analogously to example 20 with the difference that 1H-Pyrazole-3,5-dicarboxylic acid diethyl ester was used instead of 1H-Pyrazole-3,5-dicarboxylic acid diethyl ester. MS (ES+): m/e=506, chloro pattern.
This compound was isolated as a by-product in example 23.
MS (ES+): m/e=506, chloro pattern.
To a solution of 100 mg 2-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-thiophen-2-yl-2H-pyrazole-3-carboxylic acid, 0.5 ml N-NEM in 2 ml DCM, 68 mg TOTU were added and the mixture was stirred for 30 min at RT. Then 49 mg morpholine were added and the reaction was further stirred for 16 h. After the addition of 2 ml sat. NaHCO3 the mixture was filtered through a Chem Elut® cartridge by elution with ethyl acetate and then concentrated under reduced pressure. The residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. The product was obtained as its trifluoroacetate salt.
Yield: 73 mg MS (ES+): m/e=547, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that methyl-amine hydrochloride was used instead of morpholine. MS (ES+): m/e=491, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 2-amino-ethanol was used instead of morpholine. MS (ES+): m/e=521, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that amino-acetic acid hydrochloride was used instead of morpholine.
MS (ES+): m/e=535, chloro pattern.
To a solution of 50 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid, 0.1 ml N-NEM in 1 ml DCM, 34 mg TOTU were added and the mixture was stirred for 10 min at RT. Then 10 μl hydrazine hydrate were added and the reaction was further stirred for 2 h. After removal of the solvent under reduced pressure the residue was codestilled with toluene (2×10 ml) and the dissolved in 1 ml THF. Then 91 mg Carbonic acid ditrichloromethyl ester were added at RT and the reaction mixture was stirred for 48 h. After the addition of 2 ml sat. NaHCO3 the mixture was filtered through a Chem Elut® cartridge by elution with ethyl acetate and then concentrated under reduced pressure. The residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. The product was obtained as its trifluoroacetate salt.
Yield: 7 mg MS (ES+): m/e=518, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 2-piperazin-1-yl-ethanol was used instead of morpholine. MS (ES+): m/e=590, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that bis-(2-methoxy-ethyl)-amine was used instead of morpholine. MS (ES+): m/e=593, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 1-(2-hydroxy-ethyl)-imidazolidin-2-one was used instead of morpholine.
MS (ES+): m/e=590, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that methylamino-acetic acid was used instead of morpholine. MS (ES+): m/e=549, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that thiomorpholine was used instead of morpholine. MS (ES+): m/e=563, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that piperidin-4-ol was used instead of morpholine. MS (ES+): m/e=561, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that pyrrolidin-3-ol was used instead of morpholine. MS (ES+): m/e=547, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that piperidin-4-yl-methanol was used instead of morpholine. MS (ES+): m/e=575, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 8-aza-spiro[4.5]decane hydrochloride was used instead of morpholine.
MS (ES+): m/e=599, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 3-methanesulfonyl-pyrrolidine was used instead of morpholine.
MS (ES+): m/e=609, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that (1,1-Dioxo-tetrahydro-1-thiophen-3-yl)-methyl-amine was used instead of morpholine.
MS (ES+): m/e=609, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that piperazin-2-one was used instead of morpholine. MS (ES+): m/e=560, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that[1,4]oxazepane was used instead of morpholine. MS (ES+): m/e=561, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 2-trifluoromethyl-pyrrolidine was used instead of morpholine. MS (ES+): m/e=599, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 2-amino-ethanesulfonic acid amide was used instead of morpholine. MS (ES+): m/e=584, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that cyclopropylamine was used instead of morpholine. MS (ES+): m/e=517, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that cyclobutylamine was used instead of morpholine. MS (ES+): m/e=531, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 2-methoxy-ethylamine was used instead of morpholine. MS (ES+): m/e=535, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that pyrrolidine was used instead of morpholine. MS (ES+): m/e=531, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that cyanamide was used instead of morpholine. MS (ES+): m/e=502, chloro pattern.
To a solution of 5 g 5-Chloro-pyridin-2-ylamine and 1.5 ml pyridine in 30 ml toluene, 8 g bromo-acetyl bromide dissolved in 10 ml toluene was added dropwise under ice cooling. After 2 h the precipitate was isolated by filtration and recrystallized from toluene to yield a white solid.
Yield: 12 g.
The title compound was prepared analogously to example 20 with the difference that 2-Bromo-N-(5-chloro-pyridin-2-yl)-acetamide was used instead of 3-Bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole in the alkylation step. MS (ES+): m/e=463, chloro pattern.
The title compound was prepared analogously to example 21 with the difference that 2-Bromo-N-(5-chloro-pyridin-2-yl)-acetamide was used instead of 3-Bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole in the alkylation step. MS (ES+): m/e=449, chloro pattern.
Alternatively 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid can be prepared by the following procedure: To a solution of 1.5 g 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid ethyl ester in 100 ml CH2Cl2, 13.96 ml BBR3 (1M in CH2Cl2) were added and the mixture stirred at RT for 2 days. The solvent was removed in vacuo and the residue was purified by chromatography on silica gel using CH2Cl2/MeOH/HOAC/H2O=9/1/0.1/0.1. The fractions containing the product were evaporated and lyophilized. The product was obtained as its hydrobromide. Yield: 1.33 g
MS (ES+): m/e=449, chloro pattern.
The title compound was prepared analogously to example 46 with the difference that 2-Bromo-N-(5-chloro-pyridin-2-yl)-acetamide was used instead of 3-Bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole in the alkylation step. MS (ES+): m/e=502, chloro pattern.
The title compound was prepared analogously to example 1 with the difference that 3-Trifluoromethyl-1H-pyrazole-4-carboxylic acid ethyl ester was used instead of 5-Thiophen-2-yl-2H-pyrazole-3-carboxylic acid ethyl ester. MS (ES+): m/e=502, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 1-Isopropyl-piperidin-4-ylamine dihydrochloride was used instead of morpholine.
MS (ES+): m/e=602, chloro pattern.
The title compound was prepared analogously to example 1 with the difference that 4-Cyano-2H-pyrazole-3-carboxylic acid ethyl ester was used instead of 5-Thiophen-2-yl-2H-pyrazole-3-carboxylic acid ethyl ester. MS (ES+): m/e=459, chloro pattern.
This compound was isolated as a by-product in example 55. MS (ES+): m/e=459, chloro pattern.
The title compound was prepared analogously to example 1 with the difference that 2-Bromo-N-(4-chloro-phenyl)-acetamide and 4-Cyano-2H-pyrazole-3-carboxylic acid ethyl ester were used instead of 2-Bromo-N-(5-chloro-pyridin-2-yl)-acetamide and 5-Thiophen-2-yl-2H-pyrazole-3-carboxylic acid ethyl ester in the alkylation step. MS (ESI+): m/e=429, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 3-Amino-propionic acid was used instead of morpholine. MS (ES+): m/e=549, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that O-Methyl-hydroxylamine hydrochloride was used instead of morpholine.
MS (ES+): m/e=507, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 2-Amino-acetamide hydrochloride was used instead of morpholine. MS (ES+): m/e=534, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that Amino-acetic acid ethyl ester hydrochloride was used instead of morpholine.
MS (ES+): m/e=563, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 3-Amino-propan-1-ol was used instead of morpholine. MS (ES+): m/e=535, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 2S-Azetidine-2-carboxylic acid was used instead of morpholine.
MS (ES+): m/e=561, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 2S-Pyrrolidin-2-yl-methanol was used instead of morpholine.
MS (ES+): m/e=561, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 2S-Pyrrolidine-2-carboxylic acid was used instead of morpholine.
MS (ES+): m/e=575, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 2S,2-Methoxymethyl-pyrrolidine was used instead of morpholine.
MS (ES+): m/e=575, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 2R,5R,2,5-Bis-methoxymethyl-pyrrolidine was used instead of morpholine.
MS (ES+): m/e=619, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 4,5-Dihydro-oxazol-2-ylamine hydrochloride was used instead of morpholine.
MS (ES+): m/e=546, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that Piperidine-4-carboxylic acid ethyl ester was used instead of morpholine.
MS (ES+): m/e=617, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 2-Morpholin-4-yl-ethylamine was used instead of morpholine.
MS (ES+): m/e=590, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 4,4-Difluoro-piperidine hydrochloride was used instead of morpholine.
MS (ES+): m/e=581, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that Oxazolidin-2-one was used instead of morpholine. MS (ES+): m/e=547, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 1-(2-Amino-ethyl)-imidazolidin-2-one was used instead of morpholine.
MS (ES+): m/e=589, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 2,2,2-Trifluoro-ethylamine was used instead of morpholine. MS (ES+): m/e=559, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 1,1-Dioxo-tetrahydro-1-thiophen-3-ylamine was used instead of morpholine.
MS (ES+): m/e=595, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 1-(3-Amino-propyl)-pyrrolidin-2-one was used instead of morpholine.
MS (ES+): m/e=602, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that Azetidine was used instead of morpholine. MS (ES+): m/e=517, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that Thiazolidine was used instead of morpholine. MS (ES+): m/e=549, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 2-Amino-3,3,3-trifluoro-propionic acid was used instead of morpholine.
MS (ES+): m/e=603, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that C-Trimethylsilanyl-methylamine was used instead of morpholine.
MS (ES+): m/e=563, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that 1-Piperidin-4-yl-pyrrolidin-2-one hydrochloride was used instead of morpholine.
MS (ES+): m/e=628, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that Methanesulfonamide was used instead of morpholine. MS (ES+): m/e=555, chloro pattern.
The title compound was prepared analogously to example 25 with the difference that Azetidin-3-ol hydrochloride was used instead of morpholine. MS (ES+): m/e=533, chloro pattern.
To a solution of 16 g oxalic acid diethyl ester in 350 ml THF, 10.1 g KOt-Bu were added at 0° C. Then 10 g 1-furan-2-yl-ethanone in 50 ml THF were added dropwise. After 1 h the reaction mixture was diluted with 300 ml ethyl acetate and 200 ml water. This solution was acidified with diluted hydrochloric acid to pH 5. The organic layer was separated, washed with 150 ml water, dried over MgSO4, filtered and concentrated under reduced pressure to yield a white solid.
Yield: 12 g.
To a solution of 1 g [5-(5-Chloro-thiophen-2-yl)-isoxazol-3-yl]-methanol [prepared by adopting a procedure described by Ewing, William R.; Becker, Michael R.; Choi-Sledeski, Yong Mi; Pauls, Heinz W.; He, Wei; Condon, Stephen M.; Davis, Roderick S.; Hanney, Barbara A.; Spada, Alfred P.; Burns, Christopher J.; Jiang, John Z.; Li, Aiwen; Myers, Michael R.; Lau, Wan F.; Poli, Gregory B; PCT Int. Appl. (2001) 460 pp. WO 0107436 A2] and 3.01 g polymer bound triphenyl phosphine (Fluka, 3 mmol triphenylphosphine/g resin) added at 0° C. Then 2.1 g di-tert-butyl azodicarboxylate were added and the reaction mixture was stirred at RT for 2 h. The solids were filtered off and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with a n-heptane/ethyl acetate gradient 100%->50%.
Yield: 1.6 g.
A solution of 1 g N,N′-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-hydrazine dicarboxylic acid tert-butyl ester was stirred in 15 ml methanolic hydrochloric acid (8M) for 16 h at RT. Then 150 ml toluene were added and the solvents were removed under reduced pressure.
Yield: 780 mg.
A solution of 550 mg 4-Furan-2-yl-2,4-dioxo-butyric acid ethyl ester and 601 mg [5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-hydrazine in 10 ml acetic acid was heated to 80° C. for 2 h. Then the reaction mixture was diluted with 20 ml water and extracted with ethyl acetate (3×100 ml). The combined organic layers were dried over MgSO4. The solvents were removed under reduced pressure and the residue was purified by silica gel chromatography eluting with a n-heptane:ethyl acetate gradient 100%->50%.
To a solution of 400 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-furan-2-yl-1H-pyrazole-3-carboxylic acid ethyl ester in 5 ml THF and 1 ml water, 1 ml aqueous NaOH (1M) were added and the mixture was stirred for 16 h at RT. Then the solution was acidified to pH 3 with half concentrated hydrochloric acid to precipitate the pure product, which was collected by filtration. Yield: 360 mg.
To 240 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-furan-2-yl-1H-pyrazole-3-carboxylic acid in 4 ml DCM and 0.4 ml NEt3, 173 mg 1-Isopropyl-piperidin-4-ylamine dihydrochloride and 163 mg BOP-Cl were added at RT and the mixture was stirred for 16 h. After addition of 5 ml of water the mixture was filtered through a Chem Elut® cartridge by elution with ethyl acetate and then concentrated under reduced pressure. The residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. The product was obtained as its trifluoroacetate salt.
Yield: 260 mg MS (ES+): m/e=500, chloro pattern.
To a solution of 260 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-furan-2-yl-1H-pyrazole-3-carboxylic acid (1-isopropyl-piperidin-4-yl)-amide in 10 ml CCl4/MeCN/water 2:2:3, 500 mg NaIO4 and 2.1 mg Ru(III) Cl3 were added at RT. The reaction mixture was vigorously stirred for 16 h and then filtered through a Chem Elut® cartridge by elution with ethyl acetate and then concentrated under reduced pressure. The residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a brown solid. The product was obtained as its trifluoroacetate salt.
Yield: 130 mgMS (ES+): m/e=478, chloro pattern.
To a solution of 50 mg 2-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-2H-pyrazole-3-carboxylic acid, 0.1 ml N-NEM in 2 ml DCM, 34 mg TOTU and 9 mg azetidine were added and the mixture was stirred for 16 h at RT. Then, the reaction mixture was concentrated under reduced pressure and the residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. The product was obtained as its trifluoroacetate salt.
Yield: 2.7 mg MS (ES+): m/e=517, chloro pattern.
The title compound was prepared analogously to example 86 with the difference that 2-Amino-ethanesulfonic acid amide hydrochloride was used instead of azetidine.
MS (ES+): m/e=584, chloro pattern.
To a solution of 650 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid hydrochloride and 133 mg dietanolamine in 20 ml absolute DMF, 413 mg TOTU and 441 μl DIPEA were added and the mixture was stirred at RT for 3 h. The solvent was removed in vacuo and the residue purified by chromatography on silica gel using CH2Cl2/MeOH/HOAC/H2O=8/2/0.2/0.2. The fractions containing the product were evaporated and lyophilized after addition of acetic acid to give a white solid. The product was obtained as its acetate. Yield: 280 mg
MS (ES+): m/e=565, chloro pattern.
To a solution of 600 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid and 141 mg 2-Amino-2-hydroxymethyl-propane-1,3-diol in 20 ml absolute DMF, 381 mg TOTU and 407 μl DIPEA were added and the mixture was stirred at RT for 3 h. The solvent was removed in vacuo and the residue purified by chromatography on silica gel using CH2Cl2/MeOH/HOAC/H2O=8/2/0.2/0.2. The fractions containing the product were evaporated and lyophilized after addition of acetic acid to give a white solid. The product was obtained as its acetate.
Yield: 210 mg MS (ES+): m/e=581, chloro pattern.
To a solution of 800 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid hydrochloride and 239 mg L-glycine-isopropylester hydrochloride in 10 ml absolute DMF, 509 mg TOTU and 813 μl DIPEA were added and the mixture was stirred at RT for 3 h. The solvent was removed in vacuo and the residue purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized after addition of hydrochloric acid to give a white solid. The product was obtained as its hydrochloride.
Yield: 585 mg MS (ES+): m/e=577, chloro pattern.
To a solution of 10 g 1H-Pyrazole-3,5-dicarboxylic acid diethyl ester in 200 ml absolute DMF 1.885 g of a 60% suspension of NaH in mineral oil were added in an argon atmosphere. The mixture was stirred for 15 min at RT. 11.76 g 2-Bromo-N-(5-chloro-pyridin-2-yl)-acetamide were added and the mixture stirred for 2 h at RT. After concentration in vacuo the residue was purified by chromatography on silica gel using CH2Cl2/ethylacetate=8/2. The fractions containing the product were evaporated.
Yield: 16.05 g.
To a solution of 8 g 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-1H-pyrazole-3,5-dicarboxylic acid diethyl ester in 200 ml THF and 50 ml H2O 17.2 ml 1N NaOH is added. After standing for 16 h, the solution was acidified using 1 N HCl. THF was removed in vacuo and water was removed by lyophilization. The residue was purified by chromatography on silica gel using ethyl acetate followed by CH2Cl2/MeOH/HOAC/H2O=9/1/0.1/0.1. The fractions containing the product were evaporated and lyophilized to give a white solid.
Yield: 4.42 g.
To a solution of 4.42 g 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-1H-pyrazole-3,5-dicarboxylic acid 3-ethyl ester and 2.69 g 1-Isopropyl-piperidin-4-ylamine dihydrochloride in 100 ml absolute DMF, 4.1 g TOTU and 6.54 ml DIPEA were added and the mixture was stirred at RT for 4 h. Then 1.345 g 1-Isopropyl-piperidin-4-ylamine dihydrochloride, 2.05 g TOTU and 3.27 ml DIPEA were added. After standing for 16 h the solvent was removed in vacuo, the residue was dissolved in CH2Cl2 and the CH2Cl2 solution washed two times with a saturated NaHCO3 solution. The organic phase was dried over Na2SO4. After filtration and removal of the solvent in vacuo the residue was purified by chromatography on silica gel using CH2Cl2/MeOH/HOAC/H2O=9/1/0.1/0.1. The fractions containing the product were evaporated and lyophilized to give a white solid. The product was obtained as its acetate. Yield: 2.96 g MS (ES+): m/e=477, chloro pattern.
To a solution of 1.5 g 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid ethyl ester in 100 ml CH2Cl2, 13.96 ml BBR3 (1M in CH2Cl2) were added and the mixture stirred at RT for 2 days. The solvent was removed in vacuo and the residue was purified by chromatography on silica gel using CH2Cl2/MeOH/HOAC/H2O=9/1/0.1/0.1. The fractions containing the product were evaporated and lyophilized. The product was obtained as its hydrobromide.
Yield: 1.33 g.
To a solution of 400 mg 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid hydrobromide and 116 mg glycine-isopropyl ester-hydrochloride in 15 ml absolute DMF, 247 mg TOTU and 401 μl DIPEA were added and the mixture was stirred at RT for 2 h. The solvent was removed in vacuo, the residue was dissolved in CH2Cl2 and the CH2Cl2 solution washed two times with a saturated NaHCO3 solution. The organic phase was dried over Na2SO4. After filtration and removal of the solvent in vacuo the residue was purified by chromatography on silica gel using CH2Cl2/MeOH/HOAC/H2O=9/1/0.1/0.1. The fractions containing the product were evaporated and lyophilized. The residue was dissolved in CH2Cl2. Water was added and the pH of the mixture was adjusted to pH 13 by adding 1N NaOH. The phases were separated and the organic phase dried over Na2SO4. After filtration, the solvent was evaporated, the residue dissolved in water and lyophilized after addition of hydrochloric acid. The product was obtained as its hydrochloride.
Yield: 352 mg MS (ES+): m/e=548, chloro pattern.
To a solution of 400 mg 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid hydrobromide and 105 mg glycine-ethyl ester-hydrochloride in 15 ml absolute DMF, 247 mg TOTU and 401 μl DIPEA were added and the mixture was stirred at RT for 2 h. The solvent was removed in vacuo, the residue was dissolved in CH2Cl2 and the CH2Cl2 solution washed two times with a saturated NaHCO3 solution. The organic phase was dried over Na2SO4. After filtration and removal of the solvent in vacuo the residue was purified by chromatography on silica gel using CH2Cl2/MeOH/HOAC/H2O=9/1/0.1/0.1. The fractions containing the product were evaporated and lyophilized. The residue was dissolved in CH2Cl2. Water was added and the pH of the mixture was adjusted to pH 13 by adding 1N NaOH. The phases were separated and the organic phase dried over Na2SO4. After filtration, the solvent was evaporated, the residue dissolved in water and lyophilized after addition of hydrochloric acid. The product was obtained as its hydrochloride.
Yield: 352 mg MS (ES+): m/e=534, chloro pattern.
To a solution of 250 mg 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid hydrobromide and 62 mg glycine-tert.butyl ester in 10 ml absolute DMF, 154 mg TOTU and 167 μl DIPEA were added and the mixture was stirred at RT for 2 h. The solvent was removed in vacuo, the residue was dissolved in CH2Cl2 and the CH2Cl2 solution washed two times with a saturated NaHCO3 solution. The organic phase was dried over Na2SO4. After filtration and removal of the solvent in vacuo the residue was purified by chromatography on silica gel using CH2Cl2/MeOH/HOAC/H2O=9/1/0.1/0.1. The fractions containing the product were evaporated and lyophilized. The residue was dissolved in CH2Cl2. Water was added and the pH of the mixture was adjusted to pH 13 by adding 1N NaOH. The phases were separated and the organic phase dried over Na2SO4. After filtration, the solvent was evaporated and the residue dissolved in 10 ml of 90% trifluoro acetic acid. After 1 h at RT trifluoro acetic acid was removed in vacuo, the residue dissolved in water by adding CH3CN and lyophilized after addition of hydrochloric acid. The product was obtained as its hydrochloride.
Yield: 114 mg MS (ES+): m/e=506, chloro pattern.
To a solution of 500 mg 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid hydrobromide and 103 mg Azetidin-3-ol in 20 ml absolute DMF 309 mg TOTU and 501 μl DIPEA were added and the mixture was stirred at RT for 2 h. The solvent was removed in vacuo, the residue was dissolved in CH2Cl2 and the CH2Cl2 solution washed two times with a saturated NaHCO3 solution. The organic phase was dried over Na2SO4. After filtration and removal of the solvent in vacuo the residue was purified by chromatography on silica gel using CH2Cl2/MeOH/HOAC/H2O=9/1/0.1/0.1. The fractions containing the product were evaporated and lyophilized. The residue was dissolved in CH2Cl2. Water was added and the pH of the mixture was adjusted to pH 13 by adding 1N NaOH. The phases were separated and the organic phase dried over Na2SO4. After filtration, the solvent was evaporated, the residue dissolved in water and lyophilized after addition of acetic acid. The product was obtained as its acetate. Yield: 249 mg MS (ES+): m/e=504, chloro pattern.
To a solution of 400 mg 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid hydrobromide and 327 mg Cyclopropyl-methanol in 15 ml absolute DMF 171 mg Dicyclohexylcarbodiimide and 83 mg DMAP were added and the mixture was stirred at RT for 16 h. Then additional 171 mg Dicyclohexylcarbodiimide were added. After 1 day at RT the solvent was removed in vacuo and the residue was purified by chromatography on silica gel using CH2Cl2/MeOH/HOAc/H2O=9/1/0.1/0.1 and by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized. The residue was dissolved in CH2Cl2. Water was added and the pH of the mixture was adjusted to pH 13 by adding 1N NaOH. The phases were separated and the organic phase dried over Na2SO4. After filtration, the solvent was evaporated, the residue dissolved in water and lyophilized after addition of acetic acid. The product was obtained as its acetate. Yield: 92 mg MS (ES+): m/e=503, chloro pattern.
To a solution of 1.5 g 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid hydrochloride and 530 mg L-valine-isopropylester hydrochloride in 20 ml absolute DMF 954 mg TOTU and 1.524 ml DIPEA were added and the mixture was stirred at RT for 3 h. After standing for 16 h at RT, the solvent was removed in vacuo. The residue was dissolved in ethyl acetate and the solution washed with a solution of KHSO4/K2SO4 in water (2 times) and a saturated NaHCO3 solution. The phases were separated and the organic phase dried over Na2SO4. After filtration, the solvent was removed in vacuo and the residue purified by chromatography on silica gel using CH2Cl2/MeOH=100/0->40/60 and preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized after addition of hydrochloric acid to give a white solid. The product was obtained as its hydrochloride.
Yield: 1.36 g MS (ES+): m/e=605, chloro pattern.
To a solution of 760 mg 2-{[1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carbonyl]-amino}-3-methyl-butyric acid ethyl ester in 12.5 ml THF and 3.1 ml water 1.256 ml of 1N NaOH were added and the mixture stirred for 8 h at RT. The solution was diluted with water, acidified by adding HCl and lyophilized. The residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized after addition of hydrochloric acid to give a white solid. The product was obtained as its hydrochloride.
Yield: 608 mg MS (ES+): m/e=577, chloro pattern.
A mixture of 24.5 g morpholine and 13.3 g 1-Fluoro-4-nitro-benzene in 30 ml DMSO was heated to 100° C. for 4 h. This solution was poured on to 300 ml of water and the resulting precipitate was collected by filtration to yield a bright yellow crystalline product, which was dried in vacuo.
Yield: 19.7 g.
To a solution of 10 g 4-(4-Nitro-phenyl)-morpholine in 200 ml DCM, 32 g Benzyl-triethyl-ammonium chloride and 22.7 g potassium permanganate (325 mesh) were cautiously added at RT. After stirring for 1 h at RT the reaction mixture was heated to reflux for 10 h. Then a solution of 95 g Na2SO3 in 450 ml water were added under ice cooling and vigourous stirring. The mixture was filtered through a pad of celite and the filtrate was concentrated under reduced pressure. The yellow solid was stirred with 250 ml water and the precipitated product was collected by filtration. This crude product was purified by chromatography on silica gel eluting with a gradient of DCM/MeOH 100%->50%. The fractions containing the product were combined and the solvent evaporated under reduced pressure. Yield: 2.6 g.
To a solution of 2.6 g 4-(4-Nitro-phenyl)-morpholin-3-one in 350 ml ethyl acetate and 17 ml ethanol, 13.2 g SnCl2 dihydrate were added and the reaction mixture was heated to reflux for 2 h. Then after cooling to RT the mixture was stirred for 16 h. The precipitated product was collected by filtration and was pure enough for the next reaction step. Yield: 2.07 g.
To 100 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-furan-2-yl-1H-pyrazole-3-carboxylic acid in 2 ml DCM and 0.1 ml NEt3, 62 mg 4-(4-Amino-phenyl)-morpholin-3-one and 67 mg BOP-Cl were added at RT and the mixture was stirred for 16 h. The mixture was was concentrated under reduced pressure and the residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid.
Yield: 63 mg MS (ES+): m/e=550, chloro pattern.
To a solution of 4 g 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-1H-pyrazole-3,5-dicarboxylic acid 3-ethyl ester in 50 ml THF 26 ml BH3*THF (1M in THF) were added slowly at RT. Then the mixture was warmed to 40° C. for 6 h. After cooling to 0° C. 20 ml MeOH were added cautiously and the mixture was concentrated to dryness. The residue was again codistilled with 20 ml of MeOH and then purified by chromatography on silica gel eluting with n-heptane/ethyl acetate. The fractions containing the product were combined and the solvent evaporated under reduced pressure. Yield: 1.9 g.
To a solution of 380 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-hydroxymethyl-1H-pyrazole-3-carboxylic acid ethyl ester in 5 ml THF and 5 ml of water 3 ml of a 1M NaOH were added and the reaction mixture was stirred for 3 h at RT. Then the mixture was acidified with half concentrated hydrochloric acid to pH 3 and the precipitate collected by filtration and washed with 10 ml water. The product was obtained as a white solid which was dried under reduced pressure. Yield: 320 mg.
To 100 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-hydroxymethyl-1H-pyrazole-3-carboxylic acid in 2 ml DCM and 0.1 ml NEt3, 67 mg 4-(4-Amino-phenyl)-morpholin-3-one and 74 mg BOP-Cl were added at RT and the mixture was stirred for 16 h. The mixture was concentrated under reduced pressure and triturated in a mixture of water/DMF. The precipitate was collected by filtration and washed with water containing 0.5% TFA. The product was obtained as a white solid which was dried under reduced pressure.
Yield: 108 mg MS (ES+): m/e=514, chloro pattern.
To a solution of 100 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-hydroxymethyl-1H-pyrazole-3-carboxylic acid ethyl ester in 2 ml DMF 11 mg NaH (60% in mineral oil) were added at RT and stirred for 10 min. Then 100 mg 1-Bromo-2-(2-methoxy-ethoxy)-ethane were added and the mixture was stirred for 16 h. After addition of 5 ml of water the mixture was filtered through a Chem Elut® cartridge by elution with ethyl acetate and then concentrated under reduced pressure. The crude residue was directly subjected to the next reaction step. Yield: 130 mg.
To a solution of 130 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-[2-(2-methoxy-ethoxy)-ethoxymethyl]-1H-pyrazole-3-carboxylic acid ethyl ester in 5 ml THF and 5 ml of water 3 ml of a 1M NaOH were added and the reaction mixture was stirred for 3 h at RT. Then the mixture was acidified with half concentrated hydrochloric acid to pH 3 and the precipitate collected by filtration and washed with 10 ml water. The product was obtained as a white solid which was dried under reduced pressure. Yield: 60 mg.
To 60 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-[2-(2-methoxy-ethoxy)-ethoxymethyl]-1H-pyrazole-3-carboxylic acid in 2 ml DCM and 0.1 ml NEt3, 30 mg 1-Isopropyl-piperidin-4-ylamine hydrochloride and 34 mg BOP-Cl were added at RT and the mixture was stirred for 16 h. The mixture was concentrated under reduced pressure and the residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. Yield: 19 mg MS (ES+): m/e=566, chloro pattern.
To a solution of 1 g 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-1H-pyrazole-3,5-dicarboxylic acid 3-ethyl ester in 15 ml THF 314 mg LiBH4 were added cautiously. Then the reaction mixture was stirred for 16 h, quenched with diluted HCl and filtered through a Chem Elut® cartridge by elution with ethyl acetate and DCM. After concentration under reduced pressure the crude residue was directly subjected to the next reaction step.
Yield: 800 mg.
To a solution of 500 mg of 2-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-hydroxymethyl-2H-pyrazole-3-carboxylic acid in 5 ml DMF 480 mg Cs2CO3 and 204 mg 1-Bromo-2-methoxy-ethane were added and the mixture was heated to 80° C. for 5 h. Then the mixture was acidified to pH 4 with aqueous HCl and filtered through a Chem Elut® cartridge by elution with ethyl acetate. After concentration the crude product was directly subjected to the next reaction step.
To a solution of 200 mg 2-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(2-methoxy-ethoxymethyl)-2H-pyrazole-3-carboxylic acid, 0.25 ml N-NEM in 5 ml DCM, 165 mg TOTU were added and the mixture was stirred for 30 min at RT. Then 165 mg 1-Isopropyl-piperidin-4-ylamine hydrochloride were added and the reaction was further stirred for 16 h. The reaction mixture was concentrated under reduced pressure and then purified by preparative HPLC (C18 reverse phase column, elution with a H2OAMeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. The product was obtained as its trifluoroacetate salt.
Yield: 200 mg MS (ES+): m/e=523, chloro pattern.
To a solution of 2 g 2-(2-Methoxy-ethoxy)-ethanol in 20 ml THF 1.8 g KOt-Bu were added at 0° C. After stirring for 10 min 8.1 ml 3-Bromo-propyne (75% in toluene) were added and the mixture was warmed to RT and stirred for 4 h. Then 10 ml of water were added and the mixture was filtered through a Chem Elut® cartridge by elution with CHCl3. This solution containing the desired product was subjected to the next reaction step.
To a solution containing approx. 2.5 g 3-[2-(2-Methoxy-ethoxy)-ethoxy]-propyne, 2.8 g Diazo-acetic acid tert-butyl ester were added and the mixture was heated to 70° C. for 5 days. Then the solution was concentrated under reduced pressure and directly purified by chromatography on silica eluting with a n-heptane/ethyl acetate gradient. Yield: 1.7 g.
To a solution of 350 mg 5-[2-(2-Methoxy-ethoxy)-ethoxymethyl]-2H-pyrazole-3-carboxylic acid tert-butyl ester in 5 ml of DMF and 46 mg of sodium hydride (60% in mineral oil) were added at RT. After stirring for 20 min at room temperature 290 mg of 2-Bromo-N-(5-chloro-pyridin-2-yl)-acetamide were added. The reaction was stirred at room temperature for 3 h. Then 10 ml of water were added and the mixture was filtered through a Chem Elut® cartridge by elution with DCM. After concentration under reduced pressure the crude was subjected to the next reaction without further purification. Yield: 400 mg.
To a solution of 400 mg 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-[2-(2-methoxy-ethoxy)-ethoxymethyl]-1H-pyrazole-3-carboxylic acid tert-butyl ester in 5 ml DCM, 15 ml TFA were added at RT. After 3 h 30 ml toluene were added and the solvents were removed under reduced pressure. The residue was then three times codistillied with toluene and subjected to the next reaction step without further purification.
To 400 mg 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-[2-(2-methoxy-ethoxy)-ethoxymethyl]-1H-pyrazole-3-carboxylic acid in 5 ml DCM and 0.3 ml NEt3, 217 mg 1-Isopropyl-piperidin-4-ylamine hydrochloride and 250 mg BOP-Cl were added at RT and the mixture was stirred for 16 h. The mixture was concentrated under reduced pressure and the residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid.
Yield: 87 mg MS (ES+): m/e=537, chloro pattern.
To a solution of 5 g Diethyl Oxalacetate sodium salt in 100 ml ethanol, 1.5 g hydrazine monohydrochloride were added and the reaction mixture was heated to 80° C. for 3 h. Then the solution was diluted with 100 ml of water containing 3 ml of half concentrated HCl and extracted with DCM (3×100 ml). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The crude product was subjected to the next reaction step without further purification. Yield: 3.4 g
To a mixture of 3.4 g 5-Hydroxy-2H-pyrazole-3-carboxylic acid ethyl ester and 3 g K2CO3 in 50 ml acetonitrile, 4 g 1-(2-Bromo-ethoxy)-2-methoxy-ethane were added. After stirring for 1 h at RT the reaction was heated to 50° C. for 4 h. Then 50 ml of water were added and the mixture was extracted with DCM (3×100 ml). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by chromatography on silica eluting with a DCM/MeOH gradient. Yield: 1 g.
To a solution of 1 g 5-[2-(2-Methoxy-ethoxy)-ethoxy]-2H-pyrazole-3-carboxylic acid ethyl ester in 10 ml of DMF and 154 mg of sodium hydride (60% in mineral oil) were added at RT. After stirring for 5 min at room temperature, 966 mg of 2-Bromo-N-(5-chloro-pyridin-2-yl)-acetamide were added. The reaction was stirred at room temperature for 2 h. Then 50 ml of water were added and the mixture was extracted with DCM (3×100 ml). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The product was used in the next reaction step without further purification. Yield: 1.2 g.
To a solution of 1.2 g 2-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-[2-(2-methoxy-ethoxy)-ethoxy]-2H-pyrazole-3-carboxylic acid ethyl ester in 5 ml THF 10 ml of a aqueous KOH solution (10%) were added and the reaction mixture was stirred for 4 h at RT. Then the mixture was acidified with half concentrated hydrochloric acid to pH 3 and the precipitate collected by filtration and washed with 10 ml of water The product was obtained as a white solid which was dried under reduced pressure. Yield: 310 mg.
To 310 mg 2-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-[2-(2-methoxy-ethoxy)-ethoxy]-2H-pyrazole-3-carboxylic acid in 5 ml DCM and 0.3 ml NEt3, 110 mg 1-Isopropyl-piperidin-4-ylamine hydrochloride and 197 mg BOP-CL were added at RT and the mixture was stirred for 16 h. The mixture was concentrated under reduced pressure and the residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid.
Yield: 108 mg MS (ES+): m/e=523, chloro pattern.
The title compound was prepared analogously to example 104 with the difference that Trifluoro-methanesulfonic acid 2,2,2-trifluoro-ethyl ester was used instead of 1-(2-Bromo-ethoxy)-2-methoxy-ethane in step (ii). MS (ES+): m/e=503, chloro pattern.
To a solution of 600 mg 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid in 10 ml DMF, 0.9 ml 2-Methoxyethanol, 934 mg DCC and 552 mg DMAP were added. After stirring for 8 h at 40° C. the reaction mixture was directly purified by chromatography on silica gel using CH2Cl2/MeOH/HOAc/H2O=9/1/0.1/0.1 and by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized. The residue was dissolved in CH2Cl2. Water was added and the pH of the mixture was adjusted to pH 13 by adding 1N NaOH. The phases were separated and the organic phase dried over Na2SO4. After filtration, the solvent was evaporated, the residue was dissolved in water and lyophilized after addition of hydrochloric acid. The product was obtained as its hydrochloride.
Yield: 345 mg MS (ES+): m/e=507, chloro pattern.
The title compound was prepared analogously to example 106 with the difference that 10 equivalents of Ethane-1,2-diol were used instead of 2-Methoxyethanol.
MS (ES+): m/e=493, chloro pattern.
The title compound was prepared analogously to example 92 with the difference that[1,4]Oxazepane hydrochloride was used instead of glycine-isopropyl ester-hydrochloride.
MS (ES+): m/e=532, chloro pattern.
The title compound was prepared analogously to example 21 with the difference that 1-Bromomethyl-3-methoxy-benzene was used instead of 3-bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole. MS (ES+): m/e=401.
The title compound was prepared analogously to example 107 with the difference that 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid was used instead of 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid. MS (ES+): m/e=522, chloro pattern.
The title compound was prepared analogously to example 110 with the difference that Hydroxy-acetic acid tert-butyl ester was used instead of 1 Ethane-1,2-diol.
A solution of 180 mg 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid tert-butoxycarbonylmethyl ester in 20 ml TFA were allowed to stand for 20 min at RT. Then the solvent was removed under reduced pressure and the residue was dissolved in water and lyophilized after addition of hydrochloric acid. The product was obtained as its hydrochloride. Yield: 145 mg MS (ES+): m/e=536, chloro pattern.
To a solution of 10 g 1H-Pyrazole-3,5-dicarboxylic acid diethyl ester in 400 ml acetonitrile 13 g ammonium cerium(iv) nitrate (CAN) and 7.17 g iodine were added and the mixture was heated to reflux for 5 h. Then, after cooling to RT, 30 ml sat. sodium thiosulfate solution were added. The mixture was extracted with ethyl acetate (3×100 ml), the combined organic layers were washed with water and then dried over MgSO4, filtered and the solvents were removed under reduced pressure. The residue was filtered through a pad of silica gel eluting with heptane/ethyl acetate 1:1. Yield: 13 g.
To a solution of 1 g 4-Iodo-1H-pyrazole-3,5-dicarboxylic acid diethyl ester in 5 ml 2,2,2-trifluoro-ethanol 1.4 g Cs2CO3, 56 mg CuI and 106 mg 1,10-Phenanthroline were added. The reaction mixture heated for 4 h to 100° C. under microwave irradiation (100 W, CEM Discover™ apparatus). Then 10 HCl in ethanol (8M) was added and the solution was stirred at RT. After 16 h the solvents were removed under reduced pressure and the residue taken up in DCM and water. The organic phase was separated and the aqueous layer was extracted with DCM (2×50 ml). The combined organic layers were dried over MgSO4, filtered and the solvent was removed under reduced pressure. The residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. Yield: 359 mg.
To a solution of 260 mg 4-(2,2,2-Trifluoro-ethoxy)-1H-pyrazole-3,5-dicarboxylic acid diethyl ester in 4 ml absolute DMF 33.5 mg of a 60% suspension of NaH in mineral oil were added under an argon atmosphere. The mixture was stirred for 15 min at RT. Then 209 mg 2-Bromo-N-(5-chloro-pyridin-2-yl)-acetamide were added and the mixture stirred for 2 h at RT. After concentration in vacuo the residue was directly subjected to the next reaction step without further purification.
Yield: 400 mg.
To a solution of 400 mg 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-4-(2,2,2-trifluoro-ethoxy)-1H-pyrazole-3,5-dicarboxylic acid diethyl ester in 5 ml THF and 0.9 ml 1N NaOH were added. After standing for 16 h, the solution was acidified using 1 N HCl to pH 1. The precipitating product was collected by filtration and dried under reduced pressure.
Yield: 142 mg.
To 100 mg 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-4-(2,2,2-trifluoro-ethoxy)-1H-pyrazole-3,5-dicarboxylic acid 3-ethyl ester in 2 ml DCM and 0.2 ml NEt3, 128 mg 1-Isopropyl-piperidin-4-ylamine dihydrochloride and 87 mg BOP-Cl were added at RT and the mixture was stirred for 16 h. The mixture was concentrated under reduced pressure and the residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. The product was obtained as its trifluoroacetate salt.
Yield: 10 mg MS (ES+): m/e=575, chloro pattern.
A solution of 2 g Prop-2-yn-1-ol and 3 g Ethyl diazoacetate in 16 ml trichloromethane was stirred at 70° C. for 24 h. Then the solvent was removed under reduced pressure and the residue was purified on silica gel eluting with a gradient n-heptane/ethyl acetate 1:1->1:2. The fractions containing the product were collected and evaporated under reduced pressure.
Yield: 1.9 g.
To a solution of 768 mg 5-Hydroxymethyl-2H-pyrazole-3-carboxylic acid ethyl ester in 5 ml DMF, 1.9 g Imidazole and 3.2 g tert-Butyl-chloro-diphenyl-silane were added at RT and stirred for 3 h. Then 10 ml of water were added and the mixture was extracted with ethyl acetate (2×30 ml). The combined organic layers were washed with brine, dried over MgSO4 and filtered. The solids were removed under reduced pressure to yield the product as a yellow oil. Yield: 5 g.
To a solution of 5 g 5-(tert-Butyl-diphenyl-silanyloxymethyl)-2H-pyrazole-3-carboxylic acid ethyl ester in 10 ml DMF 4 g Cs2CO3 and 3 g 2-Bromo-N-(5-chloro-pyridin-2-yl)-acetamide were added and the mixture was stirred for 3 h. Then 10 ml of water were added and the mixture was extracted with ethyl acetate (2×50 ml). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. Inspection of the TLC and HPLC/MS indicated that a 1:1 mixture of the desired product together with the regioisomeric 5-(tert-Butyl-diphenyl-silanyloxymethyl)-2-[(5-chloro-pyridin-2-ylcarbamoyl)-methyl]-2H-pyrazole-3-carboxylic acid ethyl ester was present. Purification of this mixture on silica gel eluting with a gradient on n-heptane/ethylacetate yielded the desired product as the faster eluting less polar isomer.
Yield: 2 g.
To a solution of 2 g 5-(tert-Butyl-diphenyl-silanyloxymethyl)-1-[(5-chloro-pyridin-2-ylcarbamoyl)-methyl]-1H-pyrazole-3-carboxylic acid ethyl ester in 10 ml THF, 7 ml aqueous KOH solution (10%) were added at RT and the mixture was stirred for 16 h. Then the solution was acidified by addition of 10 ml half concentrated acetic acid and extracted with ethyl acetate (3×50 ml). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. Yield: 1.8 g.
To 500 mg 5-(tert-Butyl-diphenyl-silanyloxymethyl)-1-[(5-chloro-pyridin-2-ylcarbamoyl)-methyl]-1H-pyrazole-3-carboxylic acid in 10 ml DCM and 0.8 ml NEt3, 370 mg 1-Isopropyl-piperidin-4-ylamine dihydrochloride and 208 mg BOP-Cl were added at RT and the mixture was stirred for 16 h. Then 3 ml half concentrated HCl were added and the mixture was stirred for 2 h. After neutralization with of saturated aqueous NaHCO3 the mixture was extracted with ethyl acetate (2×50 ml) and DCM (1×50 ml). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC (C18 reverse phase column, elution with a H2O/MeCN gradient with 0.1% TFA). The fractions containing the product were evaporated and lyophilized to yield a white solid. The product was obtained as its trifluoroacetate salt.
Yield: 16 mg MS (ES+): m/e=435, chloro pattern.
can be prepared from 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-4-(2,2,2-trifluoro-ethoxy)-1H-pyrazole-3-carboxylic acid ethyl ester by a procedure analogous to example 21 or example 51.
can be prepared from 2,2-Difluoro-ethanol using a procedure analogous to example 112.
can be prepared from 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-4-(2,2-difluoro-ethoxy)-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid ethyl ester using a procedure analogous to example 21 or example 51.
can be prepared from 5-(tert-Butyl-diphenyl-silanyloxymethyl)-2-[(5-chloro-pyridin-2-ylcarbamoyl)-methyl]-2H-pyrazole-3-carboxylic acid ethyl ester using a procedure analogous to example 113.
can be prepared from 2-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-hydroxymethyl-2H-pyrazole-3-carboxylic acid (1-isopropyl-piperidin-4-yl)-amide using a procedure described by Q. Y. Chen et al. J. Fluorine Chem. (1989) 44, 433.
can be prepared from Trifluoro-methanesulfonic acid 2,2,2-trifluoro-ethyl ester using a procedure analogous to example 103.
can be prepared from Trifluoro-methanesulfonic acid 2,2-difluoro-ethyl ester using a procedure analogous to example 103.
can be prepared from Trifluoro-methanesulfonic acid 2,2-difluoro-3-hydroxy-propyl ester using a procedure analogous to example 103.
can be prepared from Trifluoro-methanesulfonic acid 2,2-difluoro-3-methoxy-propyl ester using a procedure analogous to example 103.
can be prepared from Trifluoro-methanesulfonic acid 3-difluoromethoxy-2,2-difluoro-propyl ester using a procedure analogous to example 103.
can be prepared from 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid using a procedure analogous to example 49.
can be prepared from 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid and Methyl-cyanamide [can be prepared adapting a procedure described by R. Niwa et al. Chem. Pharm. Bull. (1996) 44, 2314]using a procedure analogous to example 49.
can be prepared from 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid and Methyl-cyanamide [can be prepared adapting a procedure described by R. Niwa et al. Chem. Pharm. Bull. (1996) 44, 2314] using a procedure analogous to example 49.
can be prepared from 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid and 2,2,2-Trifluoro-ethyl-cyanamide [can be prepared adapting a procedure described by R. Niwa et al. Chem. Pharm. Bull. (1996) 44, 2314] using a procedure analogous to example 49.
can be prepared from 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid and 2,2,2-Trifluoro-ethyl-cyanamide [can be prepared adapting a procedure described by R. Niwa et al. Chem. Pharm. Bull. (1996) 44, 2314] using a procedure analogous to example 49.
can be prepared from 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid and 2,2-difluoro-ethyl-cyanamide [can be prepared adapting a procedure described by R. Niwa et al. Chem. Pharm. Bull. (1996) 44, 2314] using a procedure analogous to example 49.
can be prepared from 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid and 2,2-difluoro-ethyl-cyanamide [can be prepared adapting a procedure described by R. Niwa et al. Chem. Pharm. Bull. (1996) 44, 2314] using a procedure analogous to example 49.
can be prepared from 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid using a procedure analogous to example 59.
can be prepared from 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid and O,N-dimethyl-hydroxylamine [can be prepared by adapting a procedure described by M. Strasser et al. Helv. Chim. Acta (1988) 71, 1156 or P. Beak et al. J. Org. Chem. (1989) 54, 5574] using a procedure analogous to example 59.
can be prepared from 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid and O,N-dimethyl-hydroxylamine [can be prepared by adapting a procedure described by M. Strasser et al. Helv. Chim. Acta (1988) 71, 1156 or P. Beak et al. J. Org. Chem. (1989) 54, 5574] using a procedure analogous to example 59.
can be prepared from 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid and O-methyl-N-(2,2,2-trifluoro-ethyl)-hydroxylamine [can be prepared by adapting a procedure described by M. Strasser et al. Helv. Chim. Acta (1988) 71, 1156 or P. Beak et al. J. Org. Chem. (1989) 54, 5574] using a procedure analogous to example 59.
can be prepared from 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid and O-methyl-N-(2,2,2-trifluoro-ethyl)-hydroxylamine [can be prepared by adapting a procedure described by M. Strasser et al. Helv. Chim. Acta (1988) 71, 1156 or P. Beak et al. J. Org. Chem. (1989) 54, 5574] using a procedure analogous to example 59.
can be prepared from 1-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid and N-(2,2-difluoro-ethyl)-O-methyl-hydroxylamine [can be prepared by adapting a procedure described by M. Strasser et al. Helv. Chim. Acta (1988) 71, 1156 or P. Beak et al. J. Org. Chem. (1989) 54, 5574] using a procedure analogous to example 59.
can be prepared from 1-[5-(5-Chloro-thiophen-2-yl)-isoxazol-3-ylmethyl]-5-(1-isopropyl-piperidin-4-ylcarbamoyl)-1H-pyrazole-3-carboxylic acid and N-(2,2-difluoro-ethyl)-O-methyl-hydroxylamine [can be prepared by adapting a procedure described by M. Strasser et al. Helv. Chim. Acta (1988) 71, 1156 or P. Beak et al. J. Org. Chem. (1989) 54, 5574] using a procedure analogous to example 59.
can be prepared from 5-Hydroxy-2H-pyrazole-3-carboxylic acid ethyl ester using a procedure described by Q. Y. Chen et al. J. Fluorine Chem. (1989) 44, 433 and example 105.
can be prepared from 5-Hydroxy-2H-pyrazole-3-carboxylic acid ethyl ester and 3-Bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole using a procedure described by Q. Y. Chen et al. J. Fluorine Chem. (1989) 44, 433 and example 105.
can be prepared from 5-Hydroxy-2H-pyrazole-3-carboxylic acid ethyl ester and Trifluoro-methanesulfonic acid 2,2-difluoro-ethyl ester using a procedure analogous to example 105.
can be prepared from 5-Hydroxy-2H-pyrazole-3-carboxylic acid ethyl ester, 3-Bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole and Trifluoro-methanesulfonic acid
2,2-difluoro-ethyl ester using a procedure analogous to example 105.
can be prepared from 5-Hydroxy-2H-pyrazole-3-carboxylic acid ethyl ester and Trifluoro-methanesulfonic acid 2,2-difluoro-3-hydroxy-propyl ester using a procedure analogous to example 105.
can be prepared from 5-Hydroxy-2H-pyrazole-3-carboxylic acid ethyl ester, 3-Bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole and Trifluoro-methanesulfonic acid 2,2-difluoro-3-hydroxy-propyl ester using a procedure analogous to example 105.
can be prepared from 5-Hydroxy-2H-pyrazole-3-carboxylic acid ethyl ester and Trifluoro-methanesulfonic acid 2,2-difluoro-3-methoxy-propyl ester using a procedure analogous to example 105.
can be prepared from 5-Hydroxy-2H-pyrazole-3-carboxylic acid ethyl ester, 3-Bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole and Trifluoro-methanesulfonic acid 2,2-difluoro-3-methoxy-propyl ester using a procedure analogous to example 105.
can be prepared from 5-Hydroxy-2H-pyrazole-3-carboxylic acid ethyl ester and Trifluoro-methanesulfonic acid 3-difluoromethoxy-2,2-difluoro-propyl ester using a procedure analogous to example 105.
can be prepared from 5-Hydroxy-2H-pyrazole-3-carboxylic acid ethyl ester, 3-Bromomethyl-5-(5-chloro-thiophen-2-yl)-isoxazole and Trifluoro-methanesulfonic acid 3-difluoromethoxy-2,2-difluoro-propyl ester using a procedure analogous to example 105.
Pharmacological Testing
The ability of the compound of formulae I, Ib and Ic to inhibit factor Xa or factor VIIa or other enzymes like thrombin, plasmin, or trypsin can be assessed by determining the concentration of the compound of the formulae I, Ib and Ic that inhibits enzyme activity by 50%, i.e. the IC50 value, which was related to the inhibition constant Ki. Purified enzymes were used in chromogenic assays. The concentration of inhibitor that causes a 50% decrease in the rate of substrate hydrolysis was determined by linear regression after plotting the relative rates of hydrolysis (compared to the uninhibited control) versus the log of the concentration of the compound of formulae I, Ib and Ic. For calculating the inhibition constant Ki, the IC50 value was corrected for competition with substrate using the formula wherein Km is the Michaelis-Menten constant (Chen and Prusoff, Biochem.
Ki=IC50/{1+(substrate concentration/Km)}
Pharmacol. 22 (1973) 3099-3108; I. H. Segal, Enzyme Kinetics, 1975, John Wiley & Sons, New York, 100-125; which were incorporated herein by reference).
a) Factor Xa Assay
In the assay for determining the inhibition of factor Xa activity TBS-PEG buffer (50 mM Tris-HCl, pH 7.8, 200 mM NaCl, 0.05% (w/v) PEG-8000, 0.02% (w/v) NaN3) was used. The IC50 was determined by combining in appropriate wells of a Costar half-area microtiter plate 25 μl human factor Xa (Enzyme Research Laboratories, Inc.; South Bend, Ind.) in TBS-PEG; 40 μl 10% (v/v) DMSO in TBS-PEG (uninhibited control) or various concentrations of the compound to be tested diluted in 10% (v/v) DMSO in TBS-PEG; and substrate S-2765 (N(α)-benzyloxycarbonyl-D-Arg-Gly-L-Arg-p-nitroanilide; Kabi Pharmacia, Inc.; Franklin, Ohio) in TBS-PEG.
The assay was performed by pre-incubating the compound of formulae I, Ib and Ic plus enzyme for 10 min. Then the assay was initiated by adding substrate to obtain a final volume of 100 μl. The initial velocity of chromogenic substrate hydrolysis was measured by the change in absorbance at 405 nm using a Bio-tek Instruments kinetic plate reader (Ceres UV900HDi) at 25° C. during the linear portion of the time course (usually 1.5 min after addition of substrate). The enzyme concentration was 0.5 nM and substrate concentration was 140 μM.
b) Factor VIIa Assay
The inhibitory activity towards factor VIIa/tissue factor activity was determined using a chromogenic assay essentially as described previously (J. A. Ostrem et al., Biochemistry 37 (1998) 1053-1059 which was incorporated herein by reference). Kinetic assays were conducted at 25° C. in half-area microtiter plates (Costar Corp., Cambridge, Mass.) using a kinetic plate reader (Molecular Devices Spectramax 250). A typical assay consisted of 25 μl human factor VIIa and TF (5 nM and 10 nM, respective final concentration) combined with 40 μl of inhibitor dilutions in 10% DMSO/TBS-PEG buffer (50 mM Tris, 15 mM NaCl, 5 mM CaCl2, 0.05% PEG 8000, pH 8.15). Following a 15 minute preincubation period, the assay was initiated by the addition of 35 μl of the chromogenic substrate S-2288 (D-Ile-Pro-Arg-p-nitroanilide, Pharmacia Hepar Inc., 500 μM final concentration). The results (inhibition constants Ki (FXa) for inhibition of factor Xa) are shown in Table 1.
| Number | Date | Country | Kind |
|---|---|---|---|
| 02028915 | Dec 2002 | EP | regional |
| 03011308 | May 2003 | EP | regional |
This application is a continuation of application Ser. No. 12/185,388, filed Aug. 4, 2008, which is a continuation of application Ser. No. 10/744,744, filed Dec. 23, 2003, now U.S. Pat. No. 7,429,581, which claims the benefit of U.S. Provisional Application No. 60/468,905, filed May 8, 2003, and U.S. Provisional Application No. 60/507,142, filed Sep. 30, 2003.
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| Number | Date | Country | |
|---|---|---|---|
| 20100010045 A1 | Jan 2010 | US |
| Number | Date | Country | |
|---|---|---|---|
| 60468905 | May 2003 | US | |
| 60507142 | Sep 2003 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 12185388 | Aug 2008 | US |
| Child | 12562276 | US | |
| Parent | 10744744 | Dec 2003 | US |
| Child | 12185388 | US |
