Patent Application
20080200454
The present invention relates to certain carbon-linked tetrahydro-pyrazolo-pyridine compounds, pharmaceutical compositions containing them, and methods of using them for the treatment of disease states, disorders, and conditions mediated by cathepsin S activity.
Cathepsin S is one of the major cysteine proteases expressed in the lysosome of antigen presenting cells, mainly dendritic cells, B cells and macrophages. Cathepsin S is best known for its critical function in the proteolytic digestion of the invariant chain chaperone molecules, thus controlling antigen presentation to CD4+ T cells by major histocompatibility complex class II molecules or to NK1.1+ T cells via CD1 molecules. Cathepsin S also appears to participate in direct processing of exogenous antigens for presentation by MHC class II to CD4+ T cells or crosspresentation by MHC class I molecules to CD8+ T cells. In addition, cathepsin S in secreted form is implicated in degradation of extracellular matrix, which may contribute to the pathology of a number of diseases, including arthritis, atherosclerosis, and chronic obstructive pulmonary disease. Therefore, inhibition of cathepsin S is a promising target for the development of novel therapeutics for a variety of indications. For a review, see: Thurmond, R. L. et al. Curr. Opin. Invest. Drugs 2005, 6(5), 473-482.
Pyrazole inhibitors of cathepsin S were disclosed in a series of applications from Ortho-McNeil, and additional publications on this work have also appeared (See: U.S. Pat. Appl. Publ. Nos. 2002/0040020, 2003/0078419, and 2002/0040019; Wei, J. et al. Bioorg. Med. Chem. Lett. 2007, 17, 5525-5528; Grice, C. A. et al. Bioorg. Med. Chem. Lett. 2006, 16, 2209-2212; and Gustin, D. J. et al. Bioorg. Med. Chem. Lett. 2005, 15, 1678-1691). See also: Thurmond, R. L. et al. J. Pharm. Exp. Ther. 2004, 308, 268-276; and Thurmond, R. L. et al. J. Med. Chem. 2004, 47, 4799-4801). However, there remains a need for potent cathepsin S modulators with desirable pharmaceutical properties.
In one aspect the invention relates to compounds of the following Formula (I):
wherein:
In certain embodiments, the compound of Formula (I) is a compound selected from those species described or exemplified in the detailed description below.
In a further aspect, the invention relates to pharmaceutical compositions each comprising: (a) an effective amount of at least one chemical entity selected from compounds of Formula (I), and pharmaceutically acceptable salts, prodrugs, and metabolites thereof; and (b) a pharmaceutically acceptable excipient.
In another aspect, the invention is directed to a method of treating a subject suffering from or diagnosed with a disease, disorder, or medical condition mediated by cathepsin S activity, comprising administering to the subject in need of such treatment an effective amount of at least one chemical entity selected from compounds of Formula (I), and pharmaceutically acceptable salts, prodrugs, and metabolites thereof. Diseases, disorders and medical conditions that are mediated by cathepsin S activity include those referred to herein.
In another aspect, the chemical entities of the present invention are useful as cathepsin S modulators. Thus, the invention is directed to a method for modulating cathepsin S activity, including when such receptor is in a subject, comprising exposing cathepsin S to an effective amount of at least one chemical entity selected from compounds of Formula (I), pharmaceutically acceptable salts of compounds of Formula (I), pharmaceutically acceptable prodrugs of compounds of Formula (I), and pharmaceutically active metabolites of compounds of Formula (I).
Additional embodiments, features, and advantages of the invention will be apparent from the following detailed description and through practice of the invention.
For the sake of brevity, the disclosures of the publications, including patents, cited in this specification are herein incorporated by reference.
As used herein, the terms “including”, “containing” and “comprising” are used herein in their open, non-limiting sense.
The term “alkyl” refers to a saturated, straight- or branched-chain alkyl group having from 1 to 12 carbon atoms in the chain. Examples of alkyl groups include methyl (Me, which also may be structurally depicted by a bond, “/”), ethyl (Et), n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl (tBu), pentyl, isopentyl, tert-pentyl, hexyl, isohexyl, and groups that in light of the ordinary skill in the art and the teachings provided herein would be considered equivalent to any one of the foregoing examples.
The term “alkenyl” refers to a straight- or branched-chain alkenyl group having 2 to 12 carbons in the chain. Examples of alkenyl groups include vinyl, propenyl, 2-methyl-propenyl, butenyl, butadienyl, 2-methyl-butenyl, pentenyl, 2-methyl-pentenyl, 2-ethyl-pentenyl, 3-methyl-pentenyl, hexenyl, and groups that in light of the ordinary skill in the art and the teachings herein would be considered equivalent to any one of the foregoing examples.
The term “cycloalkyl” refers to a saturated or partially saturated, monocyclic, fused polycyclic, or spiro polycyclic carbocycle having from 3 to 12 ring atoms per carbocycle. Illustrative examples of cycloalkyl groups include the following entities, in the form of properly bonded moieties:
A “heterocycloalkyl” refers to a monocyclic, or fused, bridged, or spiro polycyclic ring structure that is saturated or partially saturated and has from 3 to 12 ring atoms per ring structure selected from carbon atoms and up to three heteroatoms selected from nitrogen, oxygen, and sulfur. In preferred embodiments, heterocycloalkyl rings have one or two heteroatoms. The ring structure may optionally contain up to two oxo groups on carbon or sulfur ring members. Illustrative entities, in the form of properly bonded moieties, include:
The term “heteroaryl” refers to a monocyclic, fused bicyclic, or fused polycyclic aromatic heterocycle (ring structure having ring atoms selected from carbon atoms and up to four heteroatoms selected from nitrogen, oxygen, and sulfur) having from 3 to 12 ring atoms per heterocycle. Illustrative examples of heteroaryl groups include the following entities, in the form of properly bonded moieties:
Those skilled in the art will recognize that the species of heteroaryl, cycloalkyl, and heterocycloalkyl groups listed or illustrated above are not exhaustive, and that additional species within the scope of these defined terms may also be selected.
The term “halogen” represents chlorine, fluorine, bromine, or iodine. The term “halo” represents chloro, fluoro, bromo, or iodo.
The term “substituted” means that the specified group or moiety bears one or more substituents. The term “unsubstituted” means that the specified group bears no substituents. The term “optionally substituted” means that the specified group is unsubstituted or substituted by one or more substituents. Where the term “substituted” is used to describe a structural system, the substitution is meant to occur at any valency-allowed position on the system that yields a stable chemical structure.
Any formula given herein is intended to represent compounds having structures depicted by the structural formula as well as certain variations or forms. In particular, compounds of any formula given herein may have asymmetric centers and therefore exist in different enantiomeric forms. All optical isomers and stereoisomers of the compounds of the general formula, and mixtures thereof, are considered within the scope of the formula. Thus, any formula given herein is intended to represent a racemate, one or more enantiomeric forms, one or more diastereomeric forms, one or more atropisomeric forms, and mixtures thereof. Furthermore, certain structures may exist as geometric isomers (i.e., cis and trans isomers), as tautomers, or as atropisomers. Additionally, any formula given herein is intended to represent hydrates, solvates, and polymorphs of such compounds, and mixtures thereof.
To provide a more concise description, some of the quantitative expressions given herein are not qualified with the term “about”. It is understood that, whether the term “about” is used explicitly or not, every quantity given herein is meant to refer to the actual given value, and it is also meant to refer to the approximation to such given value that would reasonably be inferred based on the ordinary skill in the art, including equivalents and approximations due to the experimental and/or measurement conditions for such given value. Whenever a yield is given as a percentage, such yield refers to a mass of the entity for which the yield is given with respect to the maximum amount of the same entity that could be obtained under the particular stoichiometric conditions. Concentrations that are given as percentages refer to mass ratios, unless indicated differently.
Reference to a chemical entity herein stands for a reference to any one of: (a) the actually recited form of such chemical entity, and (b) any of the forms of such chemical entity in the medium in which the compound is being considered when named. For example, reference herein to a compound such as R—COOH, encompasses reference to any one of, for example, R—COOH(s), R—COOH(sol), and R—COO−(sol). In this example, R—COOH(s) refers to the solid compound, as it could be for example in a tablet or some other solid pharmaceutical composition or preparation; R—COOH(sol) refers to the undissociated form of the compound in a solvent; and R—COO−(sol) refers to the dissociated form of the compound in a solvent, such as the dissociated form of the compound in an aqueous environment, whether such dissociated form derives from R—COOH, from a salt thereof, or from any other entity that yields R—COO− upon dissociation in the medium being considered. In another example, an expression such as “exposing an entity to compound of formula R—COOH” refers to the exposure of such entity to the form, or forms, of the compound R—COOH that exists, or exist, in the medium in which such exposure takes place. In this regard, if such entity is for example in an aqueous environment, it is understood that the compound R—COOH is in such same medium, and therefore the entity is being exposed to species such as R—COOH(aq) and/or R—COO−(aq), where the subscript “(aq)” stands for “aqueous” according to its conventional meaning in chemistry and biochemistry. A carboxylic acid functional group has been chosen in these nomenclature examples; this choice is not intended, however, as a limitation but it is merely an illustration. It is understood that analogous examples can be provided in terms of other functional groups, including but not limited to hydroxyl, basic nitrogen members, such as those in amines, and any other group that interacts or transforms according to known manners in the medium that contains the compound. Such interactions and transformations include, but are not limited to, dissociation, association, tautomerism, solvolysis, including hydrolysis, salvation, including hydration, protonation, and deprotonation. No further examples in this regard are provided herein because these interactions and transformations in a given medium are known by any one of ordinary skill in the art.
Any formula given herein is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds. Isotopically labeled compounds have structures depicted by the formulas given herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, and iodine, such as 2H, 3H, 11C, 13C, 14C, 15N, 18O, 17O, 33P, 33P, 35S, 18F, 36Cl, and 125I, respectively. Such isotopically labelled compounds are useful in metabolic studies (preferably with 14C), reaction kinetic studies (with, for example 2H or 3H), detection or imaging techniques [such as positron emission tomography (PET) or single-photon emission computed tomography (SPECT)] including drug or substrate tissue distribution assays, or in radioactive treatment of patients. In particular, an 18F or 11C labeled compound may be particularly preferred for PET or SPECT studies. Further, substitution with heavier isotopes such as deuterium (i.e., 2H) may afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements. Isotopically labeled compounds of this invention and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the schemes or in the examples and preparations described below by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent.
When referring to any formula given herein, the selection of a particular moiety from a list of possible species for a specified variable is not intended to define the same choice of the species for the variable appearing elsewhere. In other words, where a variable appears more than once, the choice of the species from a specified list is independent of the choice of the species for the same variable elsewhere in the formula, unless stated otherwise.
By way of a first example on substituent terminology, if substituent S1example is one of S1 and S2, and substituent S2example is one of S3 and S4, then these assignments refer to embodiments of this invention given according to the choices S1example is S1 and S2example is S3; S1example is S1 and S2example is S4; S example is S2 and S2example is S3; S1example is S2 and S2example is S4; and equivalents of each one of such choices. The shorter terminology “S1example is one of S1 and S2, and S2example is one of S3 and S4” is accordingly used herein for the sake of brevity, but not by way of limitation. The foregoing first example on substituent terminology, which is stated in generic terms, is meant to illustrate the various substituent assignments described herein. The foregoing convention given herein for substituents extends, when applicable, to any generic substituent symbol used herein.
Furthermore, when more than one assignment is given for any member or substituent, embodiments of this invention comprise the various groupings that can be made from the listed assignments, taken independently, and equivalents thereof. By way of a second example on substituent terminology, if it is herein described that substituent Sexample is one of S1, S2, and S3, this listing refers to embodiments of this invention for which Sexample is S1; Sexample is S2; Sexample is S3; Sexample is one of S1 and S2; Sexample is one of S1 and S3; Sexample is one of S2 and S3; Sexample is one of S1, S2 and S3; and Sexample is any equivalent of each one of these choices. The shorter terminology “Sexample is one of S1, S2, and S3” is accordingly used herein for the sake of brevity, but not by way of limitation. The foregoing second example on substituent terminology, which is stated in generic terms, is meant to illustrate the various substituent assignments described herein. The foregoing convention given herein for substituents extends, when applicable, to any generic substituent symbol used herein.
The nomenclature “Ci-j” with j>i, when applied herein to a class of substituents, is meant to refer to embodiments of this invention for which each and every one of the number of carbon members, from i to j including i and j, is independently realized. By way of example, the term C1-3 refers independently to embodiments that have one carbon member (C1), embodiments that have two carbon members (C2), and embodiments that have three carbon members (C3).
The term Cn-malkyl refers to an aliphatic chain, whether straight or branched, with a total number N of carbon members in the chain that satisfies n≦N≦m, with m>n.
Any disubstituent referred to herein is meant to encompass the various attachment possibilities when more than one of such possibilities are allowed. For example, reference to disubstituent -A-B-, where A≠B, refers herein to such disubstituent with A attached to a first substituted member and B attached to a second substituted member, and it also refers to such disubstituent with A attached to the second substituted member and B attached to the first substituted member.
According to the foregoing interpretive considerations on assignments and nomenclature, it is understood that explicit reference herein to a set implies, where chemically meaningful and unless indicated otherwise, independent reference to embodiments of such set, and reference to each and every one of the possible embodiments of subsets of the set referred to explicitly.
In some embodiments of Formula (I), —NR1R2 is a structure of Formula (II):
In other embodiments, —NR1R2 is a structure of Formula (III):
wherein W is O or S; and Rb5 and Rb6 are each independently H or C1-4alkyl.
In other embodiments of Formula (I), R1 and R2 taken together with the nitrogen to which they are attached form azetidine, pyrrolidine, piperidine, piperazine substituted with Ra, morpholine, or thiomorpholine, each unsubstituted or substituted with one, two, or three R substituents as described for Formula (I). In other embodiments, R1 and R2 taken together with the nitrogen to which they are attached form piperidine, piperazine substituted with Ra, or morpholine, each unsubstituted or substituted with one, two, or three R substituents as described for Formula (I). In other embodiments, R1 and R2 taken together with the nitrogen to which they are attached form 1,1-dioxo-1λ6-thiomorpholine, thiomorpholine 1-oxide, piperazinone substituted with Ra, [1,4]oxazepane, each unsubstituted or substituted with one, two, or three Rb substituents; or 2,5-diaza-bicyclo[2.2.1]heptane substituted with Ra, 2-oxa-5-aza-bicyclo[2.2.1]heptane, 2-oxa-6-aza-spiro[3.3]heptane, or hexahydro-furo[3,4-c]pyrrole, each of the latter four groups unsubstituted or substituted with one Rb substituent.
In some embodiments, Ra is H, methyl, isopropyl, acetyl, or tert-butoxycarbonyl. In other embodiments, Ra is phenyl, 2-hydroxyphenyl, 3-hydroxyphenyl, 4-hydroxyphenyl, benzoyl, pyridyl, 1-hydroxy-pyridyl, or cyclobutyl.
In some embodiments, each Rb substituent is independently OH, methyl, CF3, methoxycarbonyl, dimethylamino, acetamido, tert-butoxycarbamoyl, fluoro, or methoxy. In other embodiments, each Rb substituent is independently carbamoyl, amino, ethoxycarbonyl, carboxy, hydroxymethyl, 2-hydroxyacetylamino, methanesulfonylamino, or tert-butyl; or two R substituents on the same carbon taken together with the carbon to which they are attached form a dioxolane ring. In other embodiments, Rb is pyrrolidinyl, 2-oxo-pyrrolidinyl, or piperidinyl, each optionally substituted. In other embodiments, Rb is 2-oxo-piperidinyl, morpholinyl, 1-tert-butoxycarbonyl-piperidin-4-yl, 1-methyl-piperidin-4-yl, or 1-acetyl-piperidin-4-yl. In still other embodiments, Rb is pyrrolidin-1-yl, 2-oxo-pyrrolidin-1-yl, or 5-dimethylamino-1-methyl-1,3-dihydro-imidazo[4,5-b]pyridin-2-onyl, or two Rb substituents on the same carbon taken together with the carbon to which they are attached form 2-oxo-pyrrolidin-3-yl. In still other embodiments, Rb is phenyl or pyridyl, each optionally substituted.
In some embodiments, R3 is H or OH.
In some embodiments, R4 is —SO2CH3, —CONH2, or —COCONH2. In other embodiments, R4 is dimethylaminooxalyl, acetyl, dimethylsulfamoyl, methylcarbamoyl, dimethylcarbamoyl, 2-aminoacetyl, 2-acetoxyacetyl, 2-acetylamino-acetyl, tetrahydrofuran-2-carbonyl, or morpholine-4-carbonyl. In other embodiments, R4 is —SO2CH3.
In some embodiments, R5 is chloro or CF3. In other embodiments, R5 is chloro.
In some embodiments, each R6 is H.
In some embodiments, D is —C≡C—R7, and R7 is benzyl, phenethyl, phenpropyl, hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, butyl, phenoxymethyl, 2-methyl propyl, diethylaminomethyl, (1,1-dioxo-1λ6-thiomorpholin-4-yl)-methyl, benzamidomethyl, or (benzenesulfonamido)methyl. In other embodiments, R7 is cyclopentyl, cyclohexyl, phenyl, thiophenyl, or pyridyl, each unsubstituted or substituted with one or two Rk substituents. In other embodiments, R7 is phenyl, unsubstituted or substituted with two Rk substituents. In still other embodiments, R7 is 1H-indol-5-yl, 4-cyanomethyl-phenyl, 3-cyanomethyl-phenyl, 4-hydroxymethyl-phenyl, 3-hydroxymethyl-phenyl, 4-hydroxy-phenyl, 4-(3-carboxy-propyl)-phenyl, 4-(2-carboxy-ethyl)-phenyl, 4-(methoxycarbonyl)methyl-phenyl, 3-(methoxycarbonyl)methyl-phenyl, thiophen-2-yl, 3,4-dichloro-phenyl, 4-(4-iodo-phenoxy)-phenyl, 4-carboxymethyl-phenyl, 3-carboxymethyl-phenyl, 4-phenoxy-phenyl, 4-bromo-phenyl, 4-carboxy-phenyl, pyridin-4-yl, pyridin-3-yl, pyridin-2-yl, thiophen-3-yl, 2-methoxy-phenyl, 3-chlorophenyl, 2-chlorophenyl, 3-hydroxyphenyl, 4-chlorophenyl, 4-methylphenyl, 4-trifluoromethylphenyl, 4-fluorophenyl, 4-methoxyphenyl, 2,4-difluorophenyl, 2-trifluoromethylphenyl, 2-methylphenyl, 3-trifluoromethylphenyl, 4-amino-phenyl, phenyl, 4-(tert-butoxcarbamoyl)methyl-phenyl, benzyl, phenethyl, phenpropyl, hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, butyl, cyclohexyl, (diethylamino)methyl, (1,1-dioxo-1λ6-thiomorpholin-4-yl)methyl, 2-methylpropyl, phenoxymethyl, (benzamido)methyl, (benzenesulfonamido)methyl, 4-acetamido-phenyl, 4-aminomethyl-phenyl, 4-(methanesulfonamido)methyl-phenyl, 4-(benzenesulfonamido)methyl-phenyl, 4-(acetamido)methyl-phenyl, 4-(benzamido)methyl-phenyl, 4-(benzylamino)methyl-phenyl, 4-(4-methyl-benzylamino)methyl-phenyl, 4-(4-chloro-benzylamino)methyl-phenyl, 4-[benzyl(methyl)amino]methyl-phenyl, 4-pyrrolidin-1-ylmethyl-phenyl, 4-piperidin-1-ylmethyl-phenyl, or 1,2,3,4-tetrahydro-isoquinolin-1-yl.
In some embodiments, each Rk substituent in D is independently a methyl or ethyl group unsubstituted or substituted with OH, methoxy, fluoro, —CO2CH3, CO2H, CN, amino, tert-butoxycarbamoyl, methylsulfonamido, acetamido, pyrrolidinyl, or piperidinyl.
In still other embodiments, each Rk substituent in D is a methyl group substituted with NRrRs. In other embodiments, each Rk substituent in D is methylaminomethyl, dimethylaminomethyl, diethylaminomethyl, isobutylaminomethyl, tert-butoxycarbonylamino-methyl, (2-hydroxyethyl)aminomethyl, (3-hydroxypropyl)aminomethyl, (methoxycarbonylmethyl-amino)-methyl, (carboxymethyl-amino)-methyl, (2,2,2-trifluoroethyl-amino)-methyl, allylamino-methyl, (2-hydroxy-2-methyl-propylamino)-methyl, ethylaminomethyl, propylaminomethyl, [bis-(2-hydroxy-ethyl)-amino]-methyl, 3-hydroxy-propoxymethyl, phenylsulfonylamino-methyl, or benzoylamino-methyl. In other embodiments, each Rk substituent in D is 3,4-dihydro-1H-isoquinolin-2-ylmethyl, 1,3-dihydro-isoindol-2-ylmethyl, 4-(2-oxo-pyrrolidin-1-yl)-piperidin-1-ylmethyl, or 4-(4-hydroxy-2-oxo-pyrrolidin-1-yl)-piperidin-1-ylmethyl, morpholin-4-ylmethyl.
In still other embodiments, each Rk substituent in D is independently OH, methoxy, chloro, bromo, fluoro, CF3, CO2H, CN, amino, dimethylamino, acetylamino, methylsulfonamido, or methylsulfonyl. In other embodiments, each Rk substituent in D is phenoxy, 4-iodo-phenoxy, benzylamino, cyanomethyl-amino, benzimidazol-2-yl, phenethyl-amino, 3-(tert-butoxycarbonyl-methyl-amino)-propylcarbamoyl, 3-methylamino-propylcarbamoyl, pyrrolidine-1-carbonyl, 3-hydroxy-pyrrolidine-1-carbonyl, piperazine-1-carbonyl, [1,4]diazepane-1-carbonyl, 3-hydroxy-propylcarbamoyl, or 2-morpholin-4-yl-ethylcarbamoyl.
In further embodiments, Rk is a substituent of formula
In other embodiments, Rk is phenethylamino-methyl, cyclopropylamino-methyl, cyclobutylamino-methyl, cyclopentylamino-methyl, cyclohexylamino-methyl, cyclopropylmethylamino-methyl, benzylamino-methyl, (4-chloro-benzylamino)-methyl, (4-methanesulfonyl-benzylamino)-methyl, (2-chloro-benzylamino)-methyl, (3-chloro-benzylamino)-methyl, (2-fluoro-benzylamino)-methyl, (3-fluoro-benzylamino)-methyl, (4-fluoro-benzylamino)-methyl, (3,4-dichloro-benzylamino)-methyl, (2-methoxy-benzylamino)-methyl, (3-methoxy-benzylamino)-methyl, (4-methoxy-benzylamino)-methyl, (2-methyl-benzylamino)-methyl, (3-methyl-benzylamino)-methyl, (4-methyl-benzylamino)-methyl, (4-dimethylamino-benzylamino)-methyl, (4-isopropoxy-benzylamino)-methyl, (4-difluoromethoxy-benzylamino)-methyl, (4-amino-benzylamino)-methyl, (benzyl)-methyl-amino)-methyl, [(4-chloro-benzyl)-methyl-amino]-methyl, (1-phenyl-ethylamino)-methyl, phenylaminomethyl, [(pyridin-2-ylmethyl)-amino]-methyl, [(pyridin-3-ylmethyl)-amino]-methyl, [(pyridin-4-ylmethyl)-amino]-methyl, (2-hydroxy-1-phenyl-ethylamino)-methyl, [(methoxycarbonyl-phenyl-methyl)-amino]-methyl, [(thiophen-2-ylmethyl)-amino]-methyl, [(thiophen-3-ylmethyl)-amino]-methyl, (2-thiophen-2-yl-ethylamino)-methyl, [(3-methyl-thiophen-2-ylmethyl)-amino]-methyl, [(furan-2-ylmethyl)-amino]-methyl, [(2-trifluoromethyl-furan-3-ylmethyl)-amino]-methyl, (1,2,3,4-tetrahydro-naphthalen-1-ylamino)-methyl, indan-1-ylaminomethyl, (2-hydroxy-indan-1-ylamino)-methyl, [(thiazol-2-ylmethyl)-amino]-methyl, [(1-methyl-1H-imidazol-2-ylmethyl)-amino]-methyl, [(tetrahydro-furan-2-ylmethyl)-amino]-methyl, or [(tetrahydro-pyran-4-ylmethyl)-amino]-methyl. In still other embodiments, Rk is (pyridin-2-ylmethyl)-carbamoyl, (pyridin-3-ylmethyl)-carbamoyl, (pyridin-4-ylmethyl)-carbamoyl, benzyl-carbamoyl, (4-chlorobenzyl)-carbamoyl, (pyrrolidin-2-ylmethyl)-carbamoyl, (pyrrolidin-3-ylmethyl)-carbamoyl, 2-hydroxy-1-phenyl-ethylcarbamoyl, (morpholin-2-ylmethyl)-carbamoyl, (piperidin-3-ylmethyl)-carbamoyl, or (azetidin-3-ylmethyl)-carbamoyl. In still other embodiments, Rk is pyridin-2-ylmethoxymethyl, pyridin-3-ylmethoxymethyl, pyridin-4-ylmethoxymethyl, piperidin-4-ylmethoxymethyl, morpholin-2-ylmethoxymethyl, pyrrolidin-3-yloxymethyl, or 1-tert-butoxycarbonyl-pyrrolidin-3-yloxymethyl.
In still further embodiments, two adjacent Rk substituents taken together with the ring to which they are attached form a bicyclic fused ring system selected from the group consisting of indole, tetrahydroisoquinoline, 3,4-dihydro-2H-isoquinolin-1-one, 2,3,4,5-tetrahydro-1H-benzo[d]azepine, 2,3,4,5-tetrahydro-1H-benzo[c]azepine, 2,3-dihydro-1H-isoindole, benzimidazole, imidazole, 1H-pyrrolo[2,3-b]pyridine, and 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine, each fused ring system optionally substituted. In other embodiments, the fused ring system is substituted with methyl, isopropyl, isobutyl, 2,2,2-trifluoroethyl, hydroxymethyl, ethoxycarbonylmethyl, allyl, acetyl, —COCF3, tert-butoxycarbonyl, methoxycarbonyl, carboxy, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, pyrrolidine-1-carbonyl, piperidine-1-carbonyl, 4-methyl-piperazine-1-carbonyl, or morpholine-4-carbonyl.
In some embodiments, Rr is H or methyl.
In some embodiments, Rs is H, methyl, acetyl, or tert-butoxycarbonyl. In still other embodiments, Rr and R5 taken together with the nitrogen to which they are attached form azetidinyl, pyrrolidinyl, or piperidinyl, each unsubstituted or substituted with methyl, OH, methoxy, fluoro, or CF3.
In some embodiments, D is —CH═CH—R8 or —(CH2)2-3—R8, and R8 is phenyl, unsubstituted or substituted with one or two Rk substituents. In other embodiments, R8 is 1H-indol-5-yl, phenyl, 4-phenoxyphenyl, 3-hydroxyphenyl, 4-chlorophenyl, 4-methoxyphenyl, 2,4-difluorophenyl, 2-methylphenyl, or 4-hydroxymethyl-phenyl.
In some embodiments, D is —(CH2)3-5—R9, and R9 is OH. In other embodiments, R9 is NRnRo. In other embodiments, R9 is dimethylamino, cyclopentylamino, acetamido, or methanesulfonamido. In other embodiments, R9 is benzamido, benzenesulfonamido, or benzylsulfonamido, each unsubstituted or substituted with one or two Rk substituents. In other embodiments, R9 is pyrrolidine, piperidine, morpholine, piperazine, or azepine, each unsubstituted or substituted with methyl, OH, fluoro, or CF3. In other embodiments, R9 is OH, benzamido, methanesulfonamido, benzene-sulfonamido, benzylsulfonamido, 3,4-dichlorobenzenesulfonamido, 4-chlorobenzene-sulfonamido, 4-methylbenzenesulfonamido, 4-methoxybenzene-sulfonamido, N,N-dimethyl-sulfamoylurea, acetamido, 2-carboxybenzenesulfonamido, 2-nitrobenzene-sulfonamido, 3-chlorobenzenesulfonamido, 3-methoxybenzene-sulfonamido, 2-methylbenzene-sulfonamido, 2-chlorobenzenesulfonamido, 3-nitrobenzenesulfonamido, 3-methylbenzenesulfonamido, 3-cyanobenzenesulfonamido, 3-methanesulfonyl-benzenesulfonamido, 2-methanesulfonyl-benzenesulfonamido, pyrrolidin-1-yl, piperidin-1-yl, 3-methyl-pipieridin-1-yl, 4,4-difluoro-piperidin-1-yl, morpholin-4-yl, 4-methyl-piperazin-1-yl, azepan-1-yl, or cyclopentylamino.
In some embodiments, compounds of Formula (I) are selected from compounds of the following Formula (IV):
Other embodiments include combinations of the above variable definitions for Formula (I) and Formula (IV).
The invention includes also pharmaceutically acceptable salts of the compounds represented by Formula (I), preferably of those described above and of the specific compounds exemplified herein, and methods of treatment using such salts.
A “pharmaceutically acceptable salt” is intended to mean a salt of a free acid or base of a compound represented by Formula (I) that is non-toxic, biologically tolerable, or otherwise biologically suitable for administration to the subject. See, generally, S. M. Berge, et al., “Pharmaceutical Salts”, J. Pharm. Sci., 1977, 66:1-19, and Handbook of Pharmaceutical Salts, Properties, Selection, and Use, Stahl and Wermuth, Eds., Wiley-VCH and VHCA, Zurich, 2002. Preferred pharmaceutically acceptable salts are those that are pharmacologically effective and suitable for contact with the tissues of patients without undue toxicity, irritation, or allergic response. A compound of Formula (I) may possess a sufficiently acidic group, a sufficiently basic group, or both types of functional groups, and accordingly react with a number of inorganic or organic bases, and inorganic and organic acids, to form a pharmaceutically acceptable salt. Examples of pharmaceutically acceptable salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogen-phosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caproates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyne-1,4-dioates, hexyne-1,6-dioates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates, sulfonates, xylenesulfonates, phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, γ-hydroxybutyrates, glycolates, tartrates, methane-sulfonates, propanesulfonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates, and mandelates.
If the compound of Formula (I) contains a basic nitrogen, the desired pharmaceutically acceptable salt may be prepared by any suitable method available in the art, for example, treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, nitric acid, boric acid, phosphoric acid, and the like, or with an organic acid, such as acetic acid, phenylacetic acid, propionic acid, stearic acid, lactic acid, ascorbic acid, maleic acid, hydroxymaleic acid, isethionic acid, succinic acid, valeric acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, oleic acid, palmitic acid, lauric acid, a pyranosidyl acid, such as glucuronic acid or galacturonic acid, an alpha-hydroxy acid, such as mandelic acid, citric acid, or tartaric acid, an amino acid, such as aspartic acid or glutamic acid, an aromatic acid, such as benzoic acid, 2-acetoxybenzoic acid, naphthoic acid, or cinnamic acid, a sulfonic acid, such as laurylsulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, any compatible mixture of acids such as those given as examples herein, and any other acid and mixture thereof that are regarded as equivalents or acceptable substitutes in light of the ordinary level of skill in this technology.
If the compound of Formula (I) is an acid, such as a carboxylic acid or sulfonic acid, the desired pharmaceutically acceptable salt may be prepared by any suitable method, for example, treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary or tertiary), an alkali metal hydroxide, alkaline earth metal hydroxide, any compatible mixture of bases such as those given as examples herein, and any other base and mixture thereof that are regarded as equivalents or acceptable substitutes in light of the ordinary level of skill in this technology. Illustrative examples of suitable salts include organic salts derived from amino acids, such as glycine and arginine, ammonia, carbonates, bicarbonates, primary, secondary, and tertiary amines, and cyclic amines, such as benzylamines, pyrrolidines, piperidine, morpholine, and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum, and lithium.
The invention also relates to pharmaceutically acceptable prodrugs of the compounds of Formula (I), pharmaceutical compositions containing such pharmaceutically acceptable prodrugs, and treatment methods employing such pharmaceutically acceptable prodrugs. The term “prodrug” means a precursor of a designated compound that, following administration to a subject, yields the compound in vivo via a chemical or physiological process such as solvolysis or enzymatic cleavage, or under physiological conditions (e.g., a prodrug on being brought to physiological pH is converted to the compound of Formula (I)). A “pharmaceutically acceptable prodrug” is a prodrug that is non-toxic, biologically tolerable, and otherwise biologically suitable for administration to the subject. Illustrative procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in “Design of Prodrugs”, ed. H. Bundgaard, Elsevier, 1985.
Examples of prodrugs include compounds having an amino acid residue, or a polypeptide chain of two or more (e.g., two, three or four) amino acid residues, covalently joined through an amide or ester bond to a free amino, hydroxy, or carboxylic acid group of a compound of Formula (I). Examples of amino acid residues include the twenty naturally occurring amino acids, commonly designated by three letter symbols, as well as 4-hydroxyproline, hydroxylysine, demosine, isodemosine, 3-methylhistidine, norvalin, beta-alanine, gamma-aminobutyric acid, citrulline homocysteine, homoserine, ornithine and methionine sulfone.
Additional types of prodrugs may be produced, for instance, by derivatizing free carboxyl groups of structures of Formula (I) as amides or alkyl esters. Examples of amides include those derived from ammonia, primary C1-6alkyl amines and secondary di(C1-6alkyl) amines. Secondary amines include 5- or 6-membered heterocycloalkyl or heteroaryl ring moieties. Examples of amides include those that are derived from ammonia, C1-3alkyl primary amines, and di(C1-2alkyl)amines. Examples of esters of the invention include C1-7alkyl, C5-7cycloalkyl, phenyl, and phenyl(C1-6alkyl) esters. Preferred esters include methyl esters. Prodrugs may also be prepared by derivatizing free hydroxy groups using groups including hemisuccinates, phosphate esters, dimethylaminoacetates, and phosphoryloxymethyloxycarbonyls, following procedures such as those outlined in Adv. Drug Delivery Rev. 1996, 19, 115. Carbamate derivatives of hydroxy and amino groups may also yield prodrugs. Carbonate derivatives, sulfonate esters, and sulfate esters of hydroxy groups may also provide prodrugs. Derivatization of hydroxy groups as (acyloxy)methyl and (acyloxy)ethyl ethers, wherein the acyl group may be an alkyl ester, optionally substituted with one or more ether, amine, or carboxylic acid functionalities, or where the acyl group is an amino acid ester as described above, is also useful to yield prodrugs. Prodrugs of this type may be prepared as described in J. Med. Chem. 1996, 39, 10. Free amines can also be derivatized as amides, sulfonamides or phosphonamides. All of these prodrug moieties may incorporate groups including ether, amine, and carboxylic acid functionalities.
The present invention also relates to pharmaceutically active metabolites of compounds of Formula (I), and uses of such metabolites in the methods of the invention. A “pharmaceutically active metabolite” means a pharmacologically active product of metabolism in the body of a compound of Formula (I) or salt thereof. Prodrugs and active metabolites of a compound may be determined using routine techniques known or available in the art. See, e.g., Bertolini, et al., J. Med. Chem. 1997, 40, 2011-2016; Shan, et al., J. Pharm. Sci. 1997, 86 (7), 765-767; Bagshawe, Drug Dev. Res. 1995, 34, 220-230; Bodor, Adv. Drug Res. 1984, 13, 224-331; Bundgaard, Design of Prodrugs (Elsevier Press, 1985); and Larsen, Design and Application of Prodrugs, Drug Design and Development (Krogsgaard-Larsen, et al., eds., Harwood Academic Publishers, 1991).
The compounds of Formula (I) and their pharmaceutically acceptable salts, pharmaceutically acceptable prodrugs, and pharmaceutically active metabolites (collectively, “active agents”) of the present invention are useful in the methods of the invention.
The compounds of Formula (I) and their pharmaceutically acceptable salts, pharmaceutically acceptable prodrugs, and pharmaceutically active metabolites, whether alone or in combination, (collectively, “active agents”) of the present invention are useful as cathepsin S modulators in the methods of the invention. Such methods for modulating cathepsin S activity comprise exposing cathepsin S to an effective amount of at least one chemical entity selected from compounds of Formula (I), pharmaceutically acceptable salts of compounds of Formula (I), pharmaceutically acceptable prodrugs of compounds of Formula (I), and pharmaceutically active metabolites of compounds of Formula (I). Embodiments of this invention inhibit cathepsin S activity.
In some embodiments, the cathepsin S is in a subject with a disease, disorder, or medical condition mediated through modulation of the cathepsin S, such as those described herein. Symptoms or disease states are intended to be included within the scope of “medical conditions, disorders, or diseases.”
Accordingly, the invention relates to methods of using the active agents described herein to treat subjects diagnosed with or suffering from a disease, disorder, or condition mediated through cathepsin S activity, such as an autoimmune disease, an allergic condition, inflammation, a bowel disorder, tissue transplant rejection, pain, or cancer. Active agents according to the invention may therefore be used as immunomodulating agents, immunosuppressants, anti-allergy agents, anti-inflammatory agents, analgesics, or anti-cancer agents.
In some embodiments, an active agent of the present invention is administered to treat lupus, asthma, allergic reaction, atopic allergy, hay fever, atopic dermatitis, food allergy, rhinitis (such as allergic rhinitis and the inflammation caused by non-allergic rhinitis), skin immune system disorders (such as psoriasis), uveitis, inflammation, upper airway inflammation, Sjögren's syndrome, arthritis, rheumatoid arthritis, osteoarthritis, type I diabetes, atherosclerosis, multiple sclerosis, coeliac disease, inflammatory bowel disease (IBD), chronic obstructive pulmonary disorder (COPD), tissue transplant rejection, pain, neuropathic pain, chronic pain (such as pain due to conditions such as cancer, neuropathic pain, rheumatoid arthritis, osteoarthritis and inflammatory conditions), or cancer (and cancer-related processes such as angiogenesis, tumor growth, cell proliferation, and metastasis). In certain embodiments, an active agent of the present invention is administered to treat psoriasis, pain, multiple sclerosis, atherosclerosis, or rheumatoid arthritis.
Thus, the active agents may be used to treat subjects diagnosed with or suffering from a disease, disorder, or condition mediated through cathepsin S activity. The term “treat” or “treating” as used herein is intended to refer to administration of an active agent or composition of the invention to a subject for the purpose of effecting a therapeutic or prophylactic benefit through modulation of cathepsin S activity. Treating includes reversing, ameliorating, alleviating, inhibiting the progress of, lessening the severity of, or preventing a disease, disorder, or condition, or one or more symptoms of such disease, disorder or condition mediated through modulation of cathepsin S activity. The term “subject” refers to a mammalian patient in need of such treatment, such as a human. “Modulators” include both inhibitors and activators, where “inhibitors” refer to compounds that decrease, prevent, inactivate, desensitize or down-regulate cathepsin S expression or activity, and “activators” are compounds that increase, activate, facilitate, sensitize, or up-regulate cathepsin S expression or activity.
In treatment methods according to the invention, an effective amount of at least one active agent according to the invention is administered to a subject suffering from or diagnosed as having such a disease, disorder, or condition. An “effective amount” means an amount or dose sufficient to generally bring about the desired therapeutic or prophylactic benefit in patients in need of such treatment for the designated disease, disorder, or condition. Effective amounts or doses of the active agents of the present invention may be ascertained by routine methods such as modeling, dose escalation studies or clinical trials, and by taking into consideration routine factors, e.g., the mode or route of administration or drug delivery, the pharmacokinetics of the agent, the severity and course of the disease, disorder, or condition, the subject's previous or ongoing therapy, the subject's health status and response to drugs, and the judgment of the treating physician. An exemplary dose is in the range of from about 0.001 to about 200 mg of active agent per kg of subject's body weight per day, preferably about 0.05 to 100 mg/kg/day, or about 1 to 35 mg/kg/day, or about 0.1 to 10 mg/kg daily in single or divided dosage units (e.g., BID, TID, QID). For a 70-kg human, an illustrative range for a suitable dosage amount is from about 0.05 to about 7 g/day, or about 0.2 to about 2.5 g/day.
Once improvement of the patient's disease, disorder, or condition has occurred, the dose may be adjusted for preventative or maintenance treatment. For example, the dosage or the frequency of administration, or both, may be reduced as a function of the symptoms, to a level at which the desired therapeutic or prophylactic effect is maintained. Of course, if symptoms have been alleviated to an appropriate level, treatment may cease. Patients may, however, require intermittent treatment on a long-term basis upon any recurrence of symptoms.
In addition, the active agents of the invention may be used in combination with additional active ingredients in the treatment of the above conditions. The additional active ingredients may be coadministered separately with an active agent of Formula (I) or included with such an agent in a pharmaceutical composition according to the invention. In an exemplary embodiment, additional active ingredients are those that are known or discovered to be effective in the treatment of conditions, disorders, or diseases mediated by cathepsin S activity, such as another cathepsin S modulator or a compound active against another target associated with the particular condition, disorder, or disease. The combination may serve to increase efficacy (e.g., by including in the combination a compound potentiating the potency or effectiveness of an agent according to the invention), decrease one or more side effects, or decrease the required dose of the active agent according to the invention.
The active agents of the invention are used, alone or in combination with one or more additional active ingredients, to formulate pharmaceutical compositions of the invention. A pharmaceutical composition of the invention comprises: (a) an effective amount of at least one active agent in accordance with the invention; and (b) a pharmaceutically acceptable excipient.
A “pharmaceutically acceptable excipient” refers to a substance that is non-toxic, biologically tolerable, and otherwise biologically suitable for administration to a subject, such as an inert substance, added to a pharmacological composition or otherwise used as a vehicle, carrier, or diluent to facilitate administration of a agent and that is compatible therewith. Examples of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.
Delivery forms of the pharmaceutical compositions containing one or more dosage units of the active agents may be prepared using suitable pharmaceutical excipients and compounding techniques known or that become available to those skilled in the art. The compositions may be administered in the inventive methods by a suitable route of delivery, e.g., oral, parenteral, rectal, topical, or ocular routes, or by inhalation.
The preparation may be in the form of tablets, capsules, sachets, dragees, powders, granules, lozenges, powders for reconstitution, liquid preparations, or suppositories. Preferably, the compositions are formulated for intravenous infusion, topical administration, or oral administration.
For oral administration, the active agents of the invention can be provided in the form of tablets or capsules, or as a solution, emulsion, or suspension. To prepare the oral compositions, the active agents may be formulated to yield a dosage of, e.g., from about 0.05 to about 50 mg/kg daily, or from about 0.05 to about 20 mg/kg daily, or from about 0.1 to about 10 mg/kg daily.
Oral tablets may include the active ingredient(s) mixed with compatible pharmaceutically acceptable excipients such as diluents, disintegrating agents, binding agents, lubricating agents, sweetening agents, flavoring agents, coloring agents and preservative agents. Suitable inert fillers include sodium and calcium carbonate, sodium and calcium phosphate, lactose, starch, sugar, glucose, methyl cellulose, magnesium stearate, mannitol, sorbitol, and the like. Exemplary liquid oral excipients include ethanol, glycerol, water, and the like. Starch, polyvinyl-pyrrolidone (PVP), sodium starch glycolate, microcrystalline cellulose, and alginic acid are exemplary disintegrating agents. Binding agents may include starch and gelatin. The lubricating agent, if present, may be magnesium stearate, stearic acid or talc. If desired, the tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate to delay absorption in the gastrointestinal tract, or may be coated with an enteric coating.
Capsules for oral administration include hard and soft gelatin capsules. To prepare hard gelatin capsules, active ingredient(s) may be mixed with a solid, semi-solid, or liquid diluent. Soft gelatin capsules may be prepared by mixing the active ingredient with water, an oil such as peanut oil or olive oil, liquid paraffin, a mixture of mono and di-glycerides of short chain fatty acids, polyethylene glycol 400, or propylene glycol.
Liquids for oral administration may be in the form of suspensions, solutions, emulsions or syrups or may be lyophilized or presented as a dry product for reconstitution with water or other suitable vehicle before use. Such liquid compositions may optionally contain: pharmaceutically-acceptable excipients such as suspending agents (for example, sorbitol, methyl cellulose, sodium alginate, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel and the like); non-aqueous vehicles, e.g., oil (for example, almond oil or fractionated coconut oil), propylene glycol, ethyl alcohol, or water; preservatives (for example, methyl or propyl p-hydroxybenzoate or sorbic acid); wetting agents such as lecithin; and, if desired, flavoring or coloring agents.
The active agents of this invention may also be administered by non-oral routes. For example, compositions may be formulated for rectal administration as a suppository. For parenteral use, including intravenous, intramuscular, intraperitoneal, or subcutaneous routes, the agents of the invention may be provided in sterile aqueous solutions or suspensions, buffered to an appropriate pH and isotonicity or in parenterally acceptable oil. Suitable aqueous vehicles include Ringer's solution and isotonic sodium chloride. Such forms may be presented in unit-dose form such as ampules or disposable injection devices, in multi-dose forms such as vials from which the appropriate dose may be withdrawn, or in a solid form or pre-concentrate that can be used to prepare an injectable formulation. Illustrative infusion doses range from about 1 to 1000 μg/kg/minute of agent admixed with a pharmaceutical carrier over a period ranging from several minutes to several days.
For topical administration, the agents may be mixed with a pharmaceutical carrier at a concentration of about 0.1% to about 10% of drug to vehicle. Another mode of administering the agents of the invention may utilize a patch formulation to affect transdermal delivery.
Active agents may alternatively be administered in methods of this invention by inhalation, via the nasal or oral routes, e.g., in a spray formulation also containing a suitable carrier.
Exemplary chemical entities useful in methods of the invention will now be described by reference to illustrative synthetic schemes for their general preparation below and the specific examples that follow. Artisans will recognize that, to obtain the various compounds herein, starting materials may be suitably selected so that the ultimately desired substituents will be carried through the reaction scheme with or without protection as appropriate to yield the desired product. Alternatively, it may be necessary or desirable to employ, in the place of the ultimately desired substituent, a suitable group that may be carried through the reaction scheme and replaced as appropriate with the desired substituent. In addition, artisans will note that the various transformations described in the following Schemes may be performed in a different order than that depicted. Unless otherwise specified, the variables are as defined above in reference to Formula (I).
Abbreviations and acronyms used herein include the following:
Referring to Scheme A, the tetrahydro-pyrazolo-pyridine core structure of Formula (I) may be prepared from commercially available piperidones (X). Installation of the R4 substituent is accomplished through, for example, alkylation, acylation, sulfonylation, amide formation, or other suitable methods known in the art to provide ketones (XI). Alternatively, an amine protecting group, such as a Boc group, may be installed, and, at a later point in the synthesis, be removed (for example, by treatment with an acid such as HCl or TFA) and replaced with R4 (See Scheme F). Enamine formation according to general methods gives enamines (XII), which are then reacted with acyl chlorides, ArC(O)Cl, where Ar is a suitable substituted phenyl group, in the presence of a suitable tertiary amine base, to form enamines (XIII) or the corresponding beta-diketones, or a mixture thereof (not isolated). In situ reaction of the enamines with hydrazine generates pyrazoles (XIV).
Where Ar is a suitably substituted group as in Formula (XV), where substituent X is iodide, bromide, or trifluoromethanesulfonate (triflate), formation of alkynes (XVI) may be accomplished by any one of several methods, as depicted in Scheme B. In some embodiments, a three-step process is employed, including: 1) palladium-catalyzed coupling with a suitably protected alkyne ≡-PG, where PG is, for example, a trialkylsilyl group such as trimethylsilyl; 2) deprotection of the alkyne with, for example, a fluoride source such as TBAF; and 3) coupling with a suitable reagent R7X, where R7 is as defined for Formula (I) and substituent X is as defined above, to give alkynes (XVI). Alternatively, palladium-mediated coupling of compounds (XV) with alkynes ≡-R7 yields alkynes (XVI) in one step. In preferred embodiments, palladium-catalyzed couplings are performed in the presence of a palladium(0) catalyst such as PdCl2(PPh3)2, Pd(PPh3)4, or Pd2(dba)3, or a mixture thereof, a copper(1) salt such as copper(1) iodide, a tertiary amine base such as Et3N or DBU, in a polar aprotic solvent such as THF or DMF or a mixture thereof, at a temperature from about room temperature to the reflux temperature of the solvent. Alkynes (XVI) may then be hydrogenated or otherwise reduced using methods known in the art to provide alkanes and alkenes (XVII), where ALK is C2-5alkyl or C2-alkenyl. In another embodiment, phenyl groups (XV) where substituent X is iodide may be coupled with suitably substituted Reformatsky reagents (where substituent X is preferably Cl) in the presence of a palladium catalyst to form alkanes (XVIIIa) in one step.
One skilled in the art will recognize that the chemistry described in Scheme B may be used with alkynes other than ≡-R7 or ≡-PG or reagents other than XZn(C2-5alkyl)R8/R9 to generate other embodiments of Formula (I) or intermediates that are amenable to subsequent conversion into other embodiments of Formula (I). Examples of such conversions are shown in Scheme C. For example, where R7 is substituted with one or two Rk substituents, such substituents may be present for the couplings shown in Scheme B, or such substituents may be installed in subsequent reaction step(s).
As shown in Scheme C, protected amines (XVIII), where Y is ethynyl, ethynyl-(C1-4alkyl), or C3-5alkyl, and PG is a suitable nitrogen protecting group, such as a Boc or phthalimide group, may be protected and converted into substituted amines (XIX). Exemplary transformations include reactions with suitably substituted alkyl halides, sulfonyl chlorides, acyl chlorides, or carboxylic acids. Alternatively, ethers (XX), where PG is H or a suitable hydroxyl protecting group, may be optionally deprotected and subsequently acylated, alkylated, or activated by conversion to the corresponding iodide, bromide, chloride, tosylate, or mesylate, and then displaced with suitable nucleophiles. The intermediate alcohols may also be oxidized to the corresponding aldehydes and reacted with amines using reductive amination methods known in the art.
Two variations for the installation of the propyl amino chain are shown in Scheme D. One skilled in the art will recognize an amine protecting group, such as a Boc group, may be installed in place of the R4 shown, and, at a later point in the synthesis, be removed (for example, by treatment with an acid such as HCl or TFA) and replaced with R4. Pyrazoles (XXI) are alkylated with optionally protected reagents (XXII), where R3 is H, C1-4alkyl, —OC1-4alkyl, or a protected hydroxyl group, T is an aldehyde, a protected aldehyde, —CH2—OH, a —CH2— (protected hydroxyl) group, —CH2—Cl, or —CH2NR1R2, and LG is a suitable leaving group, such as a chloride, bromide, iodide, mesylate or tosylate, to give compounds (XXIII). Where T is a protected aldehyde (such as an acetal) or protected hydroxyl group, deprotection of (XXIII) is accomplished under general conditions. Resulting aldehydes are reacted with amines (XXIV) under reductive amination conditions, to provide propyl amines (XXV) where R3 is H, C1-4alkyl, or —OC1-4alkyl. Alcohols are converted to suitable leaving groups (LG), and displaced with amines HNR1R2, or are oxidized to the corresponding aldehydes for coupling with amines HNR1R2 by reductive amination. One skilled in the art will recognize that transformations from T to —CH2NR1R2 may be performed at any one of several later points in the synthesis. Where T is —CH2NR1R2, the alkylation step leads to compounds (XXV) directly.
Alternatively, pyrazoles (XXI) are reacted with epichlorohydrin or glycidyinosylate (each optionally as racemic mixtures or single enantiomers), in the presence of a suitable base, to give epoxides (XXVI). Epoxide opening with amines (XXIV), preferably at elevated temperatures, yields propyl amines (XXV) where R3 is OH.
In another embodiment, shown in Scheme E, addition of pyrazoles (XXI) to α,β-unsaturated nitriles (XXVI), in the presence of a suitable base, such as aq. NaOH, generates nitriles (XXVII). Reduction of the nitriles to the corresponding aldehydes (XXIII, not shown) is accomplished with a reducing agent such as DIBAL-H. Reductive amination of aldehydes (XXIII) with amines (XXIV) gives amines (XXV) as described in Scheme D.
In preferred embodiments, compounds of Formula (IV) are prepared as shown in Scheme F. N-Boc-Piperidone (XXX) is reacted with cyclic secondary amines HNR2, such as pyrrolidine, morpholine, or piperidine, in the presence of a catalytic amount of an acid catalyst such as p-toluenesulfonic acid or citric acid, in a solvent such as benzene or toluene, under dehydrating conditions (involving, for example, addition of molecular sieves or reaction at reflux temperature with a Dean-Stark trap), to form enamines (XXXI). Enamines (XXXI) are reacted with acid chlorides (XXXII, where substituent X is as defined above), prepared using methods known in the art, in the presence of a tertiary amine base such as TEA, DIPEA, or DBU, in a solvent such as CH2Cl2, DCE, or toluene, to provide enamines (XXXIII) or their corresponding beta-diketones (not shown), or a mixture thereof. Such compounds are not isolated, but are reacted directly with hydrazine, in a solvent such as MeOH or EtOH, to form pyrazoles (XXXIV).
The Boc protecting group of compounds (XXXIV) is removed using methods known in the art. In preferred embodiments, compounds (XXXIV) are treated with HCl or TFA in a solvent such as CH2Cl2 or 1,4-dioxane, to give the corresponding secondary amines (not shown, R4═H). Installation of R4 substituents other than H is accomplished using standard methods, including alkylation, acylation, amide coupling, sulfonylation, and other suitable transformations. In preferred embodiments, reaction with methanesulfonyl chloride in presence of a suitable tertiary amine base, or with oxamic acid in the presence of a coupling agent such as 1,1′-carbonyldiimidazole, provides pyrazoles (XXXV).
Alkylation of pyrazoles (XXXV) with reagents (XXXVI, where substituent X is iodo, bromo, chloro, mesylate or trifluoromethanesulfonate (triflate)), in the presence of a suitable base, gives compounds (XXXVII). Alkylations may also be performed as shown in Schemes D and E. In preferred embodiments, X in formula (XXXVI) is chloro, and reactions are performed using in DMF using Cs2CO3 as the base.
Compounds of Formula (IV) are prepared by reaction of compounds (XXXVII) under palladium-catalyzed conditions with alkynes (XXXVIII). In preferred embodiments, palladium-catalyzed couplings are performed in the presence of a palladium(0) catalyst such as PdCl2(PPh3)2, Pd(PPh3)4, or Pd2(dba)3, or a mixture thereof, a copper(1) salt such as copper(1) iodide, a tertiary amine base such as TEA, DIPEA, or DBU, in a polar aprotic solvent such as THF or DMF or a mixture thereof, at a temperature from about room temperature to the reflux temperature of the solvent. In further preferred embodiments, substituents Rk are present in reagents (XXXVIII). In alternative embodiments, suitable surrogate substituents are present, and the Rk substituents are formed in subsequent reaction steps using standard chemical transformations.
Additional preferred embodiments, including preferred solvents, temperatures, reagents, and other conditions, are provided in the Examples below.
Compounds of Formula (I) may be converted to their corresponding salts using methods described in the art. For example, an amine of Formula (I) may be treated with trifluoroacetic acid, HCl, citric acid, oxalic acid, tartaric acid, 2-oxo-butanoic acid, 2-oxo-hexanoic acid, 2-keto-glutaric acid, 2-pyrrolidone-5-carboxylic acid, or phosphoric acid in a solvent such as CH3CN, Et2O, CH2Cl2, THF, or MeOH to provide the corresponding salt form. Alternatively, compounds of Formula (I) may be converted to their corresponding tartrate salts by reaction with tartaric acid in Et2O, CH2Cl2, THF, or MeOH; to their corresponding monoethyl oxalate salts by reaction with mono- or diethyl oxalate in CH3CN; or to their corresponding 2-oxo-pentanoate salts by reaction with 2-oxo-pentanoic acid in CH3CN.
Those skilled in the art will recognize that the chemical transformations described above may be performed in a different order than that depicted in the above Schemes. Compounds prepared according to the schemes described above may be obtained as single enantiomers, diastereomers, or regioisomers, by enantio-, diastero-, or regiospecific synthesis, or by resolution. Compounds prepared according to the schemes above may alternately be obtained as racemic (1:1) or non-racemic (not 1:1) mixtures or as mixtures of diastereomers or regioisomers. Where racemic and non-racemic mixtures of enantiomers are obtained, single enantiomers may be isolated using conventional separation methods known to one skilled in the art, such as chiral chromatography, recrystallization, diastereomeric salt formation, derivatization into diastereomeric adducts, biotransformation, or enzymatic transformation. Where regioisomeric or diastereomeric mixtures are obtained, single isomers may be separated using conventional methods such as chromatography or crystallization.
The following specific examples are provided to further illustrate the invention and various preferred embodiments.
In obtaining the compounds described in the examples below and the corresponding analytical data, the following experimental and analytical protocols were followed unless otherwise indicated.
Unless otherwise stated, reaction mixtures were magnetically stirred at room temperature (rt). Where solutions are “dried,” they are generally dried over a drying agent such as Na2SO4 or MgSO4. Where mixtures, solutions, and extracts were “concentrated”, they were typically concentrated on a rotary evaporator under reduced pressure.
Microwave reactions were performed on a Personal Chemistry Emrys Optimizer. Individual reactions were heated to the desired temperature and held at that temperature for the allotted time.
Analytical HPLC retention times are reported in minutes, and were obtained on an Agilent HP-1100 instrument with a Phenomenex Luna C-18 (5 uM, 4.6×150 mm) column, with a flow rate of 1 mL/min, detection at 230, 254, and 280 nM, and a gradient of 10 to 100% CH3CN (0.05% TFA)/H2O (0.05% TFA).
Preparatory HPLC purifications were typically performed under one of the following sets of conditions:
Method A: Compounds were purified on a Phenomenex Synergi column (4 μm, 21×150 mm), with a flow rate of 25 mL/min, and solvent conditions as described for Analytical HPLC.
Method B: Compounds were injected onto a YMC column (C-18, 5 PM, 30×75 mm); with a flow rate of 30 mL/min; UV detection at 254 and 280 nM; and a gradient of 0 to 100% CH3CN (0.05% TFA)/H2O (0.05% TFA) over 21 min. The purified compounds were analyzed and tested as TFA salts following lyophilization, or as HCl salts following basic aqueous work up and treatment with dry HCl in 1,4-dioxane, Et2O, or MeOH.
Method C: Compounds were injected onto an Intersil ODS-3 column (C-18, 3 μM, 30×100 mm); with a flow rate of 90 mL/min; UV detection at 254 and 280 nM; and a gradient of 0 to 60% CH3CN/H2O (0.05% TFA) over 2 min. The purified compounds were analyzed and tested as TFA salts following lyophilization, or as HCl salts following basic aqueous work up and treatment with dry HCl in 1,4-dioxane, Et2O, or MeOH.
Method D: Compounds were injected onto an Intersil ODS-3 column (C-18, 3 μM, 30×100 mm); a flow rate of 90 mL/min; UV detection at 254 and 280 nM, and a gradient of 0 to 60% CH3CN/H2O (0.1% formic acid) over 2 min. The purified compounds were analyzed and tested as formic acid salts unless noted otherwise.
Method E: Compounds were injected onto a Phenomenex Luna column (C-18,10 μM, 50×250 mm); with a flow rate of 100 mL/min; UV detection at 254 and 280 nM; and a gradient of 0 to 100% CH3CN (0.05% TFA)/H2O (0.05% TFA) over 35 min. The purified compounds were analyzed and tested as TFA salts following lyophilization, or as HCl salts following basic aqueous work up and treatment with dry HCl in 1,4-dioxane, Et2O, or MeOH.
Method F: Compounds were injected onto a Xbridge Prep column (C-18, 5 μM, 30×100 mm); with a flow rate of 30 mL/min; UV detection at 254 and 280 nM; and a gradient of 5 to 99% CH3CN/H2O (20 mM NH4OH) over 18 min. The purified compounds were analyzed and tested as HCl salts following lyophilization and treatment with dry HCl in 1,4-dioxane, Et2O, or MeOH.
Mass spectra (MS) were obtained on an Agilent series 1100 MSD using electrospray ionization (ESI) in positive mode unless otherwise indicated. The MS data presented is the m/z found (typically [M+H]+) for the molecular ion.
Nuclear magnetic resonance (NMR) spectra were obtained on Bruker model DRX spectrometers (400, 500, or 600 MHz). The format of the 1H NMR data below is: chemical shift in ppm downfield of the tetramethylsilane reference (multiplicity, coupling constant J in Hz, integration). All 1H NMR data was acquired in CD3OD solvent unless otherwise indicated.
Chemical names were generated using ChemDraw Version 6.0.2 (CambridgeSoft, Cambridge, Mass.) or ACD/Name Version 9 (Advanced Chemistry Development, Toronto, Ontario, Canada).
A. 1-Methanesulfonyl-piperidin-4-one. To a solution of 4-piperidone monohydrate hydrochloride (90 g, 586 mmol) in CHCl3 (300 mL) and H2O (300 mL) was added K2CO3 (324 g, 2340 mmol). The slurry was cooled to 0 LC and treated with methanesulfonyl chloride (MsCl; 136 mL, 1.76 mol) by dropwise addition over a 1 h period (gas evolution was observed). The reaction mixture was allowed to stir for 72 h and was partitioned between CH2Cl2 (500 mL) and aq. NaHCO3 (500 mL). The aqueous layer was extracted with CH2Cl2 (3×200 mL). The organic layer was washed with 1% KHSO4 (250 mL), dried (Na2SO4), and concentrated to give the desired product (90.5 g, 87%) as a white solid. HPLC: Rt=2.2. MS (ESI): mass calcd. for C6H11NO3S, 178.1; m/z found, 178.1 [M+H]+. 1H NMR (CDCl3): 3.60 (t, J=6.5, 4H), 2.89 (s, 3H), 2.59 (t, J=6.3, 4H).
B. 3-(4-Chloro-3-iodo-phenyl)-5-methanesulfonyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine. To a solution of the above piperidone (10 g, 56 mmol) and p-toluenesulfonic acid (40 mg) in benzene (60 mL) was added morpholine (4.9 mL, 56 mmol). The reaction mixture was heated in a flask equipped with a condenser and a Dean-Stark trap at 90° C. for 16 h. The reaction mixture was cooled and concentrated to give the desired enamine as a beige solid, which was used without further purification. The enamine was dissolved in CH2Cl2 (40 mL), treated with TEA (9.4 mL, 67.2 mmol), and cooled to 0 LC. To this solution was added 4-chloro-3-iodobenzoyl chloride* (16.9 g, 56 mmol). The reaction mixture was allowed to warm to rt, stirred for 14 h, and then concentrated. The resulting red oil was diluted with EtOH (56 mL) and treated with hydrazine (5.34 mL, 170 mmol) at 0 LC. The resulting slurry was allowed to warm to rt and stirred for 16 h. EtOAc (120 mL) was added, and after 2 h the resulting precipitate was filtered and washed with additional EtOAc to afford the desired product as a white solid (8.80 g, 36%). HPLC: Rt=6.08. MS (ESI): mass calcd. for C13H13CIIN3O2S, 437.7; m/z found, 438.1 [M+H]+. 1H NMR (DMSO-d6): 8.05 (d, J=1.9, 1H), 7.51 (d, J=8.3, 1H), 7.43 (dd, J=8.4, 1.9, 2H), 4.30 (s, 2H), 3.36 (t, J=5.8, 2H), 3.30 (br s, 1H), 2.86 (s, 3H), 2.69 (t, J=5.6, 2H).
*4-Chloro-3-iodobenzoyl chloride was prepared by dissolving 4-chloro-3-iodobenzoic acid (15.8 g, 56 mmol) in CH2Cl2 (40 mL) and treating with oxalyl chloride (4.1 mL, 46.7 mmol) and a catalytic amount of DMF (400 μL; vigorous gas evolution). The mixture was stirred at rt for 3 h. The reaction mixture was concentrated to afford a white solid, which was used without further purification.
C. 3-(4-Chloro-3-iodo-phenyl)-1-(2-[1,3]dioxolan-2-yl-ethyl)-5-methanesulfonyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine. A slurry of the above pyrazole (10 g, 22.8 mmol) and Cs2CO3 (11.9 g, 45.6 mmol) in DMF (75 mL) was stirred at rt for 2 h. 2-(2-Bromoethyl)1,3-dioxolane (3.5 mL, 34.2 mmol) was added dropwise and stirring maintained for 2 days. Ice water was added slowly to form a precipitate. The white solid was collected by suction filtration and washed with H2O and Et2O to afford the desired product (10.4 g, 85%). HPLC: Rt=6.98. MS (ESI): mass calcd. for C18H21CIIN3O4S, 537.8; m/z found, 538.2 [M+H]+. 1H NMR (CDCl3): 8.15 (s, 1H), 7.46-7.45 (m, 2H), 4.83 (t, J=4.6, 1H), 4.49 (s, 2H), 4.17 (t, J=7.1, 2H), 4.01-3.97 (m, 2H), 3.89-3.86 (m, 2H), 3.65 (t, J=5.8, 2H), 2.89 (s, 3H), 2.87 (t, J=5.8, 2H), 2.28-2.26 (m, 2H).
D. 3-(4-Chloro-3-iodo-phenyl)-5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine. A mixture of the above acetal (4.65 g, 8.64 mmol) and 1 N HCl (19 mL) in acetone (75 mL) was heated at 55 LC for 5 h. The clear solution was concentrated to remove acetone, and the aqueous layer was extracted with CH2Cl2 (3×). The combined organic extracts were dried (Na2SO4), filtered, and concentrated to give a white solid, which was used directly in the next reaction. The crude aldehyde was dissolved in CH2Cl2 (80 mL) and morpholine (2.5 mL, 28.6 mmol) and acetic acid (1.0 mL) were added sequentially. After 10 min, NaB(OAc)3H (3.48 g, 13 mmol) was added and stirring was continued for 2.5 days. After the addition of 1 N NaOH, the layers were separated and the aqueous layer was extracted with CH2Cl2 (3×). The combined organic extracts were washed with brine, dried (Na2SO4), filtered, and concentrated to give an orange oil. Purification (SiO2; 0-3% 2 M NH3 in MeOH/CH2Cl2) afforded the title compound as a white solid (2.9 g, 60%). HPLC: Rt=4.78. MS (ESI): mass calcd. for C20H26CIIN4O3S, 564.9; m/z found, 566.2 [M+H]+. 1H NMR (CDCl3): 8.15 (s, 1H), 7.46-7.45 (m, 2H), 4.49 (s, 2H), 4.09 (t, J=6.8, 2H), 3.70 (t, J=4.6, 4H), 3.64 (t, J=5.8, 2H), 2.90 (s, 3H), 2.89 (t, J=5.8, 2H), 2.44-2.39 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.03 (m, 2H).
E. 3-(4-Chloro-3-trimethylsilanylethynyl-phenyl)-5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine. To a solution of the above iodoarene (1.0 g, 1.8 mmol), PdCl2(PPh)3 (124 mg, 0.177 mmol), and CuI (34 mg, 0.177 mmol) in degassed THF (10 mL) was added TEA (1.2 mL, 8.85 mmol), followed by (trimethylsilyl)acetylene (TMSA; 276 μL). The reaction mixture was stirred under N2 at rt for 17 h. Satd. aq. NaHCO3 was added, and the aqueous layer was extracted with CH2Cl2 (3×). The combined organic extracts were washed with brine, dried (Na2SO4), filtered, and concentrated to give a brown oil. Purification (SiO2; 0-3% 2 M NH3 in MeOH/CH2Cl2) afforded the desired product as a beige solid (860 mg, 91%). HPLC: Rt=5.45. MS (ESI): mass calcd. for C25H35ClN4O3SSi, 535.2; m/z found, 536.3 [M+H]+. 1H NMR (CDCl3): 7.73 (d, J=2.0, 1H), 7.46 (dd, J=8.4, 2.1, 1H), 7.40 (d, J=8.2, 1H), 4.51 (s, 2H), 4.09 (t, J=6.8, 2H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 2.90 (s, 3H), 2.89 (t, J=5.7, 2H), 2.43-2.38 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.10-2.03 (m, 2H), 0.29 (s, 9H).
A. 3-(4-Chloro-3-ethynyl-phenyl)-5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine. To a solution of 3-(4-chloro-3-trimethylsilanylethynyl-phenyl)-5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine (947 mg, 1.77 mmol) in THF (7 mL) was added tetrabutylammonium fluoride (1.0 M in THF; 2.0 mL, 1.95 mmol) dropwise. The resulting solution was stirred at rt for 30 min, then diluted with H2O. The aqueous layer was extracted with CH2Cl2 (3×). The combined organic extracts were washed with brine, dried (Na2SO4), filtered, and concentrated to give a black oil. Purification (SiO2; 0-3% 2 M NH3 in MeOH/CH2Cl2) provided the title compound as a yellow oil (550 mg, 67%). HPLC: Rt=4.62. MS (ESI): mass calcd. for C22H27ClN4O3S, 463.0; m/z found, 464.3 [M+H]+. 1H NMR (CDCl3): 7.75 (d, J=2.1, 1H), 7.52 (dd, J=8.4, 2.2, 1H), 7.43 (d, J=8.4, 1H), 4.49 (s, 2H), 4.09 (t, J=6.9, 2H), 3.71 (t, J=4.6, 4H), 3.64 (t, J=5.7, 2H), 2.91 (s, 3H), 2.88 (t, J=5.7, 2H), 2.47-2.42 (br m, 4H), 2.36 (t, J=7.1, 2H), 2.11-2.00 (m, 3H).
B. 3-[4-Chloro-3-(1H-indol-5-ylethynyl)-phenyl]-5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]oyridine. To a solution of the above alkyne (120 mg, 0.259 mmol), 5-iodoindole (82 mg, 0.337 mmol), PdCl2(PPh3)2 (18 mg, 0.026 mmol), and CuI (4.9 mg, 0.026 mmol) in degassed THF (2.4 mL) was added TEA (108 μL, 0.777 mmol). The reaction mixture was stirred at rt under N2 for 16 h. The mixture was concentrated and the resulting black oil was purified (SiO2; 0-5% 2 M NH3 in MeOH/CH2Cl2) to provide the title compound as a white solid (132 mg, 88%). HPLC: Rt=4.73. MS (ESI): mass calcd. for C30H32ClN5O3S, 578.1; m/z found, 579.3 [M+H]+. 1H NMR (CDCl3): 8.60 (s, 1H), 7.92-7.90 (m, 1H), 7.80 (t, J=1.2, 1H), 7.44 (d, J=1.2, 2H), 7.40 (dd, J=8.4, 1.5, 1H), 7.35 (d, J=8.4, 1H), 7.23-7.20 (m, 1H), 6.56-6.54 (m, 1H), 4.50 (s, 2H), 4.07 (t, J=6.8, 2H), 3.69 (t, J=4.6, 4H), 3.60 (t, J=5.7, 2H), 2.88 (s, 3H), 2.83 (t, J=5.7, 2H), 2.41-2.37 (br m, 4H), 2.31 (t, J=6.8, 2H), 2.08-2.01 (m, 2H).
A pressure tube containing a suspension of 3-[4-chloro-3-(1H-indol-5-ylethynyl)-phenyl]-5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine (45 mg, 0.079 mmol) and PtO2 (8.5 mg) in 1:1 EtOH/EtOAc (4 mL) was placed in a shaker under H2 (20 psi). After 17 h, the reaction mixture was filtered through diatomaceous earth and concentrated to give a brown oil. Purification (SiO2; 0-3% 2 M NH3 in MeOH/CH2Cl2) provided the title compound as a white powder (39 mg, 85%). HPLC: Rt=5.10. MS (ESI): mass calcd. for C30H36ClN5O3S, 582.2; m/z found, 583.5 [M+H]+. 1H NMR (CDCl3): 8.25 (br s, 1H), 7.52-7.45 (m, 2H), 7.40 (d, J=8.2, 1H), 7.35-7.31 (m, 2H), 7.19-7.16 (m, 1H), 7.07 (dd, J=8.3, 1.5, 1H), 6.51-6.48 (m, 1H), 4.39 (s, 2H), 4.08 (t, J=6.8, 2H), 3.69 (t, J=4.6, 4H), 3.62 (t, J=5.8, 2H), 3.13-3.08 (m, 2H), 3.06-3.00 (m, 2H), 2.91-2.84 (m, 2H), 2.84 (s, 3H), 2.44-2.38 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.02 (m, 2H).
The compounds in Examples 3-14 were prepared according to the methods described for Intermediate 1 and Example 1, substituting the appropriate iodide for 5-iodoindole in Example 1, Step B.
HPLC: Rt=4.81. MS (ESI): mass calcd. for C30H32ClN5O3S, 578.1; m/z found, 579.4 [M+H]+. 1H NMR (CDCl3): 7.81 (d, J=1.9, 1H), 7.62-7.59 (m, 2H), 7.50 (dd, J=8.4, 2.1, 2H), 7.34 (d, J=8.4, 2H), 4.54 (s, 2H), 4.10 (t, J=6.8, 2H), 3.79 (s, 2H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 2.91 (s, 3H), 2.90-2.87 (m, 2H), 2.43-2.38 (br m, 4H), 2.32 (t, J=6.8, 2H).
HPLC: Rt=5.43. MS (ESI): mass calcd. for C30H32ClN5O3S, 578.1; m/z found, 579.3 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=2.0, 1H), 7.59-7.55 (m, 2H), 7.53-7.33 (m, 4H), 4.55 (s, 2H), 4.11 (t, J=6.8, 2H), 3.78 (s, 2H), 3.70 (t, J=4.6, 4H), 3.66 (t, J=5.8, 2H), 2.91 (s, 3H), 2.90 (t, J=5.6, 2H), 2.43-2.39 (br m, 4H), 2.33 (t, J=6.8, 2H), 2.12-2.04 (m, 2H).
HPLC: Rt=4.82. MS (ESI): mass calcd. for C29H33ClN4O4S, 569.1; m/z found, 570.3 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=1.8, 1H), 7.61-7.57 (m, 2H), 7.51-7.44 (m, 2H), 7.37 (d, J=8.3, 2H), 4.73 (s, 2H), 4.54 (s, 2H), 4.10 (t, J=6.8, 2H), 3.69 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 2.90 (s, 3H), 2.90-2.87 (m, 2H), 2.43-2.38 (m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.04 (m, 2H), 1.87 (br s, 1H).
MS (ESI): mass calcd. for C29H33ClN4O4S, 569.1; m/z found, 570.3 [M+H]+.
HPLC: Rt=4.61. MS (ESI): mass calcd. for C28H31ClN4O4S, 555.1; m/z found, 556.4 [M+H]+. 1H NMR (CDCl3): 7.77-7.75 (m, 1H), 7.42-7.37 (m, 4H), 6.81-6.77 (m, 2H), 4.50 (s, 2H), 4.11 (t, J=6.8, 2H), 3.70 (t, J=4.5, 4H), 3.61 (t, J=5.3, 2H), 2.91 (s, 3H), 2.86 (t, J=5.6, 2H), 2.46-2.41 (m, 4H), 2.37 (t, J=6.9, 2H), 2.12-2.04 (m, 2H).
HPLC: Rt=4.89. MS (ESI): mass calcd. for C32H37ClN4O5S, 625.2; m/z found, 626.5 [M+H]+. 1H NMR (CDCl3): 7.77 (s, 1H), 7.52 (d, J=8.0, 2H), 7.47 (s, 2H), 7.21 (d, J=8.0, 2H), 4.52 (s, 2H), 4.11 (t, J=6.7, 2H), 3.74 (t, J=4.2, 4H), 3.68-3.63 (m, 4H), 3.10 (dd, J=14.6, 7.3, 1H), 2.93 (s, 3H), 2.92-2.88 (m, 2H), 2.69 (t, J=7.6, 2H), 2.53-2.47 (br m, 3H), 2.41 (t, J=7.0, 2H), 2.33 (t, J=7.4, 2H), 2.14-2.06 (m, 2H), 2.00-1.91 (m, 2H).
HPLC: Rt=4.64. MS (ESI): mass calcd. for C31H35ClN4O5S, 611.2; m/z found, 612.4 [M+H]+. 1H NMR (CDCl3): 7.70 (s, 1H), 7.45 (d, J=7.5, 2H), 7.38 (d, J=8.4, 2H), 7.22 (d, J=7.8, 2H), 6.68 (br s, 1H), 4.50 (s, 2H), 4.01 (t, J=6.5, 2H), 3.77-3.71 (m, 4H), 3.61-3.56 (m, 2H), 2.97 (t, J=7.1, 2H), 2.88 (s, 3H), 2.78-2.73 (m, 2H), 2.65 (t, J=7.1, 2H), 2.61-2.54 (m, 4H), 2.49 (t, J=6.7, 2H), 2.07 (t, J=6.6, 2H).
MS (ESI): mass calcd. for C31H35ClN4O5S, 611.2; m/z found, 612.3 [M+H]+.
MS (ESI): mass calcd. for C31H35ClN4O5S, 611.2; m/z found, 612.3 [M+H]+.
HPLC: Rt=4.91. MS (ESI): mass calcd. for C26H29ClN4O3S2, 545.1; m/z found, 546.3 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=2.0, 1H), 7.49 (dd, J=8.4, 2.0, 1H), 7.45 (d, J=8.4, 1H), 7.37-7.33 (m, 2H), 7.04 (dd, J=5.2, 3.7, 1H), 4.53 (s, 2H), 4.13-4.08 (m, 2H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 2.91 (s, 3H), 2.88 (t, J=5.8, 2H), 2.44-2.38 (br m, 4H), 2.33 (t, J=6.8, 2H), 2.11-2.03 (m, 2H).
MS (ESI): mass calcd. for C28H29Cl3N4O3S, 608.0; m/z found, 609.2 [M+H]+.
MS (ESI): mass calcd. for C34H34CIIN4O4S, 757.1; m/z found, 758.2 [M+H]+.
To a solution of (4-{2-chloro-5-[5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-phenylethynyl}-phenyl)-acetic acid methyl ester (19 mg, 0.031 mmol) in 2:1 EtOH/H2O (0.9 mL) was added NaOH (3.7 mg, 0.093 mmol). The reaction mixture was stirred at rt for 30 min and then purified directly by reverse-phase HPLC (0.05% TFA/10-98% MeCN/H2O) to afford the title compound as an off-white solid (11.7 mg, 53%). HPLC: Rt=4.75. MS (ESI): mass calcd. for C30H33ClN4O5S, 597.1; m/z found, 598.3 [M+H]+. 1H NMR (CDCl3): 7.74 (d, J=1.6, 1H), 7.56-7.53 (m, 2H), 7.46-7.43 (m, 2H), 7.32-7.29 (m, 2H), 4.50 (s, 2H), 4.12 (t, J=6.5, 2H), 3.95 (br m, 4H), 3.65-3.63 (m, 4H), 3.62 (br s, 1H), 3.15-3.09 (m, 2H), 2.92 (s, 3H), 2.82 (t, J=5.6, 4H), 2.39-2.31 (br m, 4H).
The title compound was prepared from (3-{2-chloro-5-[5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-phenylethynyl}-phenyl)-acetic acid methyl ester according to the method described in Example 15. MS (ESI): mass calcd. for C30H33ClN4O5S, 597.1; m/z found, 598.3 [M+H]
Formic acid (12 μL) was added to a degassed solution of 3-{4-chloro-3-[4-(4-iodo-phenoxy)-phenylethynyl]-phenyl}-5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine (119 mg, 0.157 mmol), Pd(OAc)2 (3.6 mg, 0.016 mmol), PPh3 (8.2 mg, 0.032 mmol), and TEA (66 μL, 0.473 mmol) in DMF (1.5 mL). The reaction mixture was heated at 60° C. under N2 for 1.5 h. After cooling to rt, saturated (satd.) aq. NaHCO3 was added, and the aqueous layer was extracted with CH2Cl2 (3×). The combined organic extracts were dried (Na2SO4), filtered, and concentrated to give a brown oil. Purification (SiO2; 0-3% 2 M NH3 in MeOH/CH2Cl2) afforded the title compound as a white solid (51 mg, 52%). MS (ESI): mass calcd. for C34H35ClN4O4S, 631.2; m/z found, 632.3 [M+H]+.
The compounds in Example 18-54 were prepared using methods analogous to those described for Intermediate 1, substituting the appropriate alkyne for TMSA in Step E.
HPLC: Rt=5.61. MS (ESI): mass calcd. for C28H30BrClN4O3S, 618.0; m/z found, 619.2 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=1.9, 1H), 7.53-7.44 (m, 6H), 4.54 (s, 2H), 4.10 (t, J=6.7, 2H), 3.70 (t, J=4.4, 4H), 3.66 (t, J=5.7, 2H), 2.91 (s, 3H), 2.91-2.88 (m, 2H), 2.32 (t, J=6.6, 2H).
HPLC: Rt=4.66. MS (ESI): mass calcd. for C29H31ClN4O5S, 583.1; m/z found, 584.4 [M+H]+. 1H NMR (CDCl3): 8.06 (d, J=8.2, 2H), 7.80 (s, 1H), 7.67 (d, J=8.2, 2H), 7.54-7.50 (m, 2H), 4.54 (s, 2H), 4.28-4.18 (m, 2H), 4.06-4.00 (br m, 4H), 3.60 (t, J=5.7, 2H), 3.24-3.13 (m, 4H), 3.08-3.01 (m, 2H), 2.98 (s, 3H), 2.82-2.77 (m, 2H).
HPLC: Rt=4.02. MS (ESI): mass calcd. for C27H30ClN5O3S, 540.1; m/z found, 541.3 [M+H]+. 1H NMR (CDCl3): 8.65-8.62 (m, 2H), 7.83 (d, J=2.0, 1H), 7.54 (dd, J=8.4, 2.1, 1H), 7.50-7.44 (m, 3H), 4.54 (s, 2H), 4.11 (t, J=6.8, 2H), 3.70 (t, J=4.5, 4H), 3.66 (t, J=5.8, 2H), 2.91 (s, 3H), 2.92-2.88 (m, 2H), 2.44-2.39 (m, 4H), 2.33 (t, J=6.8, 2H), 2.12-2.04 (m, 2H).
HPLC: Rt=4.34. MS (ESI): mass calcd. for C27H30ClN5O3S, 540.1; m/z found, 541.3 [M+H]+. 1H NMR (CDCl3): 8.83 (s, 1H), 8.58 (d, J=3.9, 1H), 7.88 (dt, J=1.9, 7.9, 1H), 7.83 (d, J=2.0, 1H), 7.52 (dd, J=8.4, 2.1, 1H), 7.47 (d, J=8.4, 1H), 7.34-7.29 (m, 1H), 4.54 (s, 2H), 4.10 (t, J=6.8, 2H), 3.69 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 2.91 (s, 3H), 2.90 (t, J=5.8, 2H), 2.43-2.38 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.04 (m, 2H).
HPLC: Rt=4.50. MS (ESI): mass calcd. for C27H30ClN5O3S, 540.1; m/z found, 541.3 [M+H]+. 1H NMR (CDCl3): 8.65 (d, J=4.5, 1H), 7.86 (d, J=2.1, 1H), 7.72 (dt, J=1.7, 7.7, 1H), 7.63-7.60 (m, 1H), 7.56 (dd, J=8.4, 2.1, 1H), 7.47 (d, J=8.4, 1H), 7.30-7.26 (m, 1H), 4.52 (s, 2H), 4.09 (t, J=6.8, 2H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 2.92 (s, 3H), 2.89 (t, J=5.7, 2H), 2.43-2.39 (br m, 4H), 2.33 (t, J=6.8, 2H), 2.11-2.03 (m, 2H).
HPLC: Rt=5.15. MS (ESI): mass calcd. for C26H29ClN4O3S2, 545.1; m/z found, 546.3 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.9, 1H), 7.61 (dd, J=3.0, 1.1, 1H), 7.49 (dd, J=8.4, 2.1, 1H), 7.44 (d, J=8.4, 1H), 7.33 (dd, J=5.0, 3.0, 1H), 7.25 (dd, J=5.0, 1.1, 1H), 4.53 (s, 2H), 4.09 (t, J=6.8, 1H), 3.69 (t, J=4.5, 4H), 3.65 (t, J=5.8, 2H), 2.90 (s, 3H), 2.89 (t, J=5.7, 2H), 2.42-2.38 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.10-2.03 (m, 2H).
HPLC: Rt=5.18. MS (ESI): mass calcd. for C29H33ClN4O4S, 569.1; m/z found, 570.3 [M+H]+. 1H NMR (CDCl3): 7.83 (d, J=1.5, 1H), 7.56 (dd, J=7.6, 1.7, 1H), 7.47-7.44 (m, 2H), 7.36-7.31 (m, 1H), 6.98-6.91 (m, 2H), 4.52 (s, 2H), 4.09 (t, J=6.8, 2H), 3.93 (s, 3H), 3.70 (t, J=4.6, 4H), 3.64 (t, J=5.8, 2H), 2.90 (s, 3H), 2.88 (t, J=5.7, 2H), 2.44-2.38 (br m, 4H), 2.33 (t, J=6.8, 2H), 2.11-2.03 (m, 2H).
HPLC: Rt=5.60. MS (ESI): mass calcd. for C28H30Cl2N4O3S, 573.6; m/z found, 574.3 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=2.0, 1H), 7.60-7.57 (m, 1H), 7.53-7.45 (m, 3H), 7.36-7.28 (m, 2H), 4.54 (s, 2H), 4.10 (t, J=6.8, 2H), 3.70 (t, J=4.6, 4H), 3.66 (t, J=5.8, 2H), 2.91 (s, 3H), 2.90 (t, J=5.7, 2H), 2.43-2.39 (br m, 4H), 2.33 (t, J=6.8, 2H), 2.12-2.05 (m, 2H).
HPLC: Rt=5.48. MS (ESI): mass calcd. for C28H30Cl2N4O3S, 573.6; m/z found, 574.3 [M+H]+. 1H NMR (CDCl3): 7.85 (d, J=1.8, 1H), 7.65-7.62 (m, 1H), 7.50-7.43 (m, 3H), 7.32-7.27 (m, 2H), 4.53 (s, 2H), 4.15-4.08 (m, 2H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 2.91-2.87 (m, 2H), 2.90 (s, 3H), 2.43-2.39 (br m, 4H), 2.33 (t, J=6.8, 2H), 2.11-2.04 (m, 2H).
HPLC: Rt=4.80. MS (ESI): mass calcd. for C28H31ClN4O4S, 555.1; m/z found, 556.6 [M+H]+. 1H NMR (CDCl3): 7.69 (d, J=2.1, 1H), 7.45 (dd, J=8.5, 2.1, 1H), 7.36 (d, J=8.4, 1H), 7.18 (t, J=7.9, 1H), 7.10-7.06 (m, 1H), 6.88-6.87 (m, 1H), 6.81-6.78 (m, 1H), 4.47 (s, 2H), 4.07 (t, J=6.7, 2H), 3.71 (t, J=4.5, 4H), 3.54 (t, J=5.7, 2H), 2.88 (s, 3H), 2.77 (t, J=5.5, 2H), 2.48-2.43 (br m, 4H), 2.38 (t, J=7.1, 2H), 2.09-2.01 (m, 2H).
HPLC: Rt=5.56. MS (ESI): mass calcd. for C28H30Cl2N4O3S, 573.6; m/z found, 574.3 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=2.0, 1H), 7.54-7.48 (m, 2H), 7.46 (d, J=8.4, 2H), 7.37-7.33 (m, 2H), 4.53 (s, 2H), 4.10 (t, J=6.8, 2H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 2.90 (s, 3H), 2.89 (t, J=5.9, 2H), 2.42-2.38 (br m, 4H), 2.32 (t, J=6.9, 2H), 2.11-2.04 (m, 2H).
MS (ESI): mass calcd. for C29H33ClN4O3S, 553.1; m/z found, 554.6 [M+H]+.
MS (ESI): mass calcd. for C29H30ClF3N4O3S, 607.1; m/z found, 608.3 [M+H]+.
MS (ESI): mass calcd. for C28H30ClFN4O3S, 557.1; m/z found, 558.3 [M+H]+.
MS (ESI): mass calcd. for C29H33ClN4O4S, 569.1; m/z found, 570.3 [M+H]+.
MS (ESI): mass calcd. for C28H29ClF2N4O3S, 575.1; m/z found, 576.3 [M+H]+.
MS (ESI): mass calcd. for C29H30ClF3N4O3S, 607.1; m/z found, 608.3 [M+H]+.
MS (ESI): mass calcd. for C29H33ClN4O3S, 553.1; m/z found, 554.6 [M+H]+.
MS (ESI): mass calcd. for C29H30ClF3N4O3S, 607.1; m/z found, 608.3 [M+H]+.
MS (ESI): mass calcd. for C29H31ClN4O4S, 567.1; m/z found, 568.3 [M+H]+.
HPLC: Rt=4.47. MS (ESI): mass calcd. for C28H32ClN5O3S, 554.1; m/z found, 555.4 [M+H]+. 1H NMR (CDCl3): 7.74 (s, 1H), 7.43-7.36 (m, 4H), 6.64 (d, J=8.5, 2H), 4.50 (s, 2H), 4.09 (t, J=6.9, 2H), 3.93 (br s, 2H), 3.72 (t, J=4.3, 4H), 3.62 (t, J=5.6, 2H), 2.89 (s, 3H), 2.88-2.84 (m, 2H), 2.50-2.45 (br m, 4H), 2.39 (t, J=6.8, 2H), 2.14-2.06 (m, 2H).
HPLC: Rt=5.19. MS (ESI): mass calcd. for C28H31ClN3O3S, 539.1; m/z found, 540.4 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=1.7, 1H), 7.61-7.59 (m, 2H), 7.47-7.44 (m, 2H), 7.38-7.36 (m, 3H), 4.52 (s, 2H), 4.10 (t, J=6.8, 2H), 3.69 (t, J=4.4, 4H), 3.64 (t, J=5.7, 2H), 2.89-2.87 (m, 5H), 2.40-2.39 (m, 4H), 2.32 (t, J=1.7, 2H), 2.08-2.05 (m, 2H).
MS (ESI): mass calcd. for C34H42ClN5O5S, 668.3; m/z found, 669.6 [M+H]+.
HPLC: Rt=5.20. MS (ESI): mass calcd. for C29H33ClN4O3S, 553.1; m/z found, 554.4 [M+H]+. 1H NMR (CDCl3): 7.71 (d, J=1.9, 1H), 7.49-7.40 (m, 4H), 7.38-7.33 (m, 2H), 7.28-7.23 (m, 1H), 4.49 (s, 2H), 4.07 (t, J=6.8, 2H), 3.92 (s, 2H), 3.69 (t, J=4.4, 4H), 3.62 (t, J=5.7, 2H), 2.87 (s, 3H), 2.86 (t, J=5.4, 2H), 2.44-2.39 (br m, 4H), 2.33 (t, J=6.8, 2H), 2.10-2.03 (m, 2H).
HPLC: Rt=5.35. MS (ESI): mass calcd. for C30H35ClN4O3S, 567.2; m/z found, 568.3 [M+H]+. 1H NMR (CDCl3): 7.63 (d, J=2.1, 1H), 7.47-7.44 (m, 2H), 7.39 (d, J=8.4, 1H), 7.33-7.30 (m, 3H), 7.25-7.20 (m, 1H), 4.50 (s, 2H), 4.09 (t, J=6.8, 2H), 3.69 (t, J=4.5, 4H), 3.64 (t, J=5.8, 2H), 2.98 (t, J=7.5, 2H), 2.89 (s, 3H), 2.88 (t, J=5.7, 2H), 2.78 (t, J=7.5, 2H), 2.43-2.38 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.10-2.03 (m, 2H).
HPLC: Rt=5.58. MS (ESI): mass calcd. for C31H37ClN4O3S, 581.2; m/z found, 582.4 [M+H]+. 1H NMR (CDCl3): 7.67 (d, J=2.0, 1H), 7.45 (dd, J=8.4, 2.1, 1H), 7.40 (d, J=8.4, 1H), 7.22-7.17 (m, 1H), 4.49 (s, 2H), 4.08 (t, J=6.8, 2H), 3.68 (t, J=4.6, 4H), 3.62 (t, J=5.8, 2H), 2.88-2.82 (m, 4H), 2.87 (s, 3H), 2.49 (t, J=6.9, 2H), 2.41-2.37 (br m, 4H), 2.31 (t, J=6.8, 2H), 2.09-2.01 (m, 2H), 2.01-1.92 (m, 2H).
MS (ESI): mass calcd. for C23H29ClN4O4S, 493.0; m/z found, 494.4 [M+H]+.
MS (ESI): mass calcd. for C24H31ClN4O4S, 507.1; m/z found, 508.4 [M+H]+.
MS (ESI): mass calcd. for C25H33ClN4O4S, 521.1; m/z found, 522.4 [M+H]+.
HPLC: Rt=5.33. MS (ESI): mass calcd. for C26H35ClN4O3S, 519.1; m/z found, 520.4 [M+H]+. 1H NMR (CDCl3): 7.65 (d, J=2.0, 1H), 7.43 (dd, J=8.4, 2.1, 1H), 7.39 (d, J=8.4, 1H), 4.50 (s, 2H), 4.08 (t, J=6.8, 2H), 3.69 (t, J=4.6, 4H), 3.64 (t, J=5.8, 2H), 2.89 (s, 3H), 2.88 (t, J=5.8, 2H), 2.49 (t, J=7.0, 2H), 2.42-2.38 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.09-2.02 (m, 2H), 1.68-1.60 (m, 2H), 1.58-1.48 (m, 2H), 0.96 (t, J=7.3, 3H).
HPLC: Rt=5.61. MS (ESI): mass calcd. for C28H37ClN4O3S, 545.2; m/z found, 546.4 [M+H]+. 1H NMR (CDCl3): 7.67-7.65 (m, 1H), 7.41-7.40 (m, 2H), 4.51 (s, 2H), 4.09 (t, J=6.8, 2H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 2.89 (s, 3H), 2.88 (t, J=5.8, 2H), 2.73-2.66 (m, 1H), 2.44-2.38 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.10-2.03 (m, 2H), 1.94-1.86 (m, 2H), 1.84-1.75 (m, 2H), 1.66-1.52 (m, 3H), 1.44-1.35 (m, 3H).
HPLC: Rt=3.98. MS (ESI): mass calcd. for C27H38ClN5O3S, 548.2; m/z found, 549.4 [M+H]+. 1H NMR (CDCl3): 7.61 (d, J=2.0, 1H), 7.39 (dd, J=8.4, 2.1, 1H), 7.34 (d, J=8.4, 1H), 4.43 (s, 2H), 4.02 (t, J=6.8, 2H), 3.66 (s, 2H), 3.62 (t, J=4.6, 4H), 3.57 (t, J=5.8, 2H), 2.82 (s, 3H), 2.81 (t, J=5.9, 2H), 2.61 (q, J=7.2, 4H), 2.35-2.31 (br m, 4H), 2.25 (t, J=6.8, 2H), 2.03-1.96 (m, 2H), 1.07 (t, J=7.2, 6H).
MS (ESI): mass calcd. for C27H36ClN5O5S2, 610.2; m/z found, 611.3 [M+H]+.
MS (ESI): mass calcd. for C26H35ClN4O3S, 519.1; m/z found, 520.4 [M+H]+.
MS (ESI): mass calcd. for C29H33ClN4O4S, 569.1; m/z found, 570.3 [M+H]+.
MS (ESI): mass calcd. for C30H34ClN5O4S, 596.2; m/z found, 597.4 [M+H]+.
MS (ESI): mass calcd. for C29H34ClN5O5S2, 632.2; m/z found, 633.4 [M+H]+.
The compounds in Examples 55-65 were prepared using methods analogous to those described in Example 2. The alkynes used as starting materials for Examples 55-65 are described in the preceding examples.
MS (ESI): mass calcd. for C34H39ClN4O4S, 635.2; m/z found, 636.4 [M+H]+.
MS (ESI): mass calcd. for C28H35ClN4O4S, 559.1; m/z found, 560.4 [M+H]+.
HPLC: Rt=5.89. MS (ESI): mass calcd. for C28H34Cl2N4O3S, 577.6; m/z found, 578.3 [M+H]+. 1H NMR (CDCl3): 7.42-7.38 (m, 2H), 7.33 (dd, J=8.3, 2.1, 1H), 7.27-7.23 (m, 2H), 7.17-7.13 (m, 2H), 4.44 (s, 2H), 4.10 (t, J=6.8, 2H), 3.70 (t, J=4.6, 2H), 3.65 (t, J=5.8, 2H), 3.06-3.01 (m, 2H), 2.94-2.87 (m, 4H), 2.89 (s, 3H), 2.43-2.37 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.10-2.03 (m, 2H).
HPLC: Rt=5.22. MS (ESI): mass calcd. for C29H37ClN4O4S, 573.2; m/z found, 574.4 [M+H]+. 1H NMR (CDCl3): 7.46 (d, J=2.0, 1H), 7.39 (d, J=8.3, 1H), 7.33 (dd, J=8.3, 2.1, 1H), 7.18-7.14 (m, 2H), 6.87-6.83 (m, 2H), 4.45 (s, 2H), 4.10 (t, J=6.8, 2H), 3.79 (s, 3H), 3.70 (t, J=4.6, 4H), 3.64 (t, J=5.8, 2H), 3.06-3.01 (m, 2H), 2.91-2.87 (m, 4H), 2.88 (s, 3H), 2.44-2.40 (br m, 4H), 2.34 (t, J=6.9, 2H), 2.11-2.03 (m, 2H).
HPLC: Rt=5.80. MS (ESI): mass calcd. for C28H33ClF2N4O3S, 579.1; m/z found, 580.3 [M+H]+. 1H NMR (CDCl3): 7.43 (d, J=2.0, 1H), 7.39 (d, J=8.3, 1H), 7.33 (dd, J=8.3, 2.1, 1H), 7.15-7.08 (m, 1H), 6.82-6.76 (m, 2H), 4.45 (s, 2H), 4.10 (t, J=6.8, 2H), 3.71 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 3.06-3.01 (m, 2H), 2.97-2.91 (m, 2H), 2.90-2.86 (m, 2H), 2.89 (s, 3H), 2.45-2.41 (br m, 4H), 2.35 (t, J=6.9, 2H), 2.11-2.05 (m, 2H).
HPLC: Rt=5.40. MS (ESI): mass calcd. for C29H37ClN4O3S, 557.2; m/z found, 558.5 [M+H]+. 1H NMR (CDCl3): 7.49 (d, J=2.0, 1H), 7.40 (d, J=8.3, 1H), 7.33 (dd, J=8.3, 2.1, 1H), 7.22-7.11 (m, 3H), 4.44 (s, 2H), 4.09 (t, J=6.8, 2H), 3.69 (t, J=4.6, 4H), 3.63 (t, J=5.8, 2H), 3.04-2.99 (m, 2H), 2.96-2.86 (m, 4H), 2.87 (s, 3H), 2.43-2.38 (br m, 4H), 2.36 (s, 3H), 2.32 (t, J=6.6, 2H), 2.10-2.03 (m, 2H).
HPLC: Rt=4.70. MS (ESI): mass calcd. for C29H37ClN4O4S, 573.2; m/z found, 574.2 [M+H]+. 1H NMR (CDCl3): 7.39 (d, J=8.2, 1H), 7.37-7.32 (m, 2H), 7.27 (d, J=8.1, 2H), 7.19 (d, J=8.0, 2H), 4.63 (s, 2H), 4.39 (s, 2H), 4.07 (t, J=6.8, 2H), 3.67 (t, J=4.6, 4H), 3.63 (t, J=5.8, 2H), 3.10-3.02 (m, 2H), 2.98-2.92 (m, 2H), 2.89-2.83 (m, 2H), 2.86 (s, 3H), 2.45 (br s, 1H), 2.42-2.36 (br m, 4H), 2.31 (t, J=6.9, 2H), 2.10-2.01 (m, 2H).
HPLC: Rt=5.24. MS (ESI): mass calcd. for C28H35ClN4O3S, 543.1; m/z found, 544.5 [M+H]+. 1H NMR (CDCl3): 7.48 (d, J=2.0, 1H), 7.39 (d, J=8.3, 1H), 7.32 (dd, J=8.1, 2.2, 2H), 7.31-7.18 (m, 5H), 4.44 (s, 2H), 4.09 (t, J=6.7, 2H), 3.69 (t, J=4.6, 4H), 3.64 (t, J=5.7, 2H), 3.09-3.04 (m, 2H), 2.97-2.92 (m, 2H), 2.89-2.86 (m, 2H), 2.87 (s, 3H), 2.43-2.38 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.10-2.03 (m, 2H).
HPLC: Rt=5.42. MS (ESI): mass calcd. for C29H37ClN4O3S, 557.2; m/z found, 558.3 [M+H]+. 1H NMR (CDCl3): 7.51 (d, J=2.0, 1H), 7.36 (d, J=8.3, 1H), 7.31-7.26 (m, 4H), 7.23-7.17 (m, 2H), 4.49 (s, 2H), 4.09 (t, J=6.8, 2H), 3.69 (t, J=4.6, 4H), 3.64 (t, J=5.8, 2H), 2.90-2.85 (m, 2H), 2.86 (s, 3H), 2.84-2.78 (m, 2H), 2.72 (t, J=7.8, 2H), 2.42-2.38 (m, 4H), 2.32 (t, J=6.8, 2H), 2.09-1.95 (m, 4H).
MS (ESI): mass calcd. for C30H38ClN5O4S, 600.2; m/z found, 601.5 [M+H]+.
HPLC: Rt=4.82. MS (ESI): mass calcd. for C29H38ClN5O5S2, 636.2; m/z found, 637.4 [M+H]+. 1H NMR (CDCl3): 7.87-7.84 (m, 2H), 7.59-7.54 (m, 1H), 7.52-7.47 (m, 2H), 7.44 (d, J=1.7, 1H), 7.37-7.31 (m, 2H), 4.87 (t, J=6.1, 1H), 4.51 (s, 2H), 4.09 (t, J=6.8, 2H), 3.69 (t, J=4.6, 4H), 3.64 (t, J=5.8, 2H), 2.99 (q, J=6.5, 2H), 2.91 (s, 3H), 2.89 (t, J=5.8, 2H), 2.79-2.74 (m, 2H), 2.43-2.38 (m, 4H), 2.32 (t, J=6.8, 2H), 2.10-2.03 (m, 2H), 1.86-1.78 (m, 2H).
To a solution of 4-{2-chloro-5-[5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-phenylethynyl}-phenylamine (80 mg, 0.144 mmol) and pyridine (28 μL, 0.347 mmol) in CH2Cl2 (1.4 mL) was added Ac2O (17 μL, 0.173 mmol) dropwise. The reaction mixture was stirred at rt for 14 h and transferred directly to a silica gel column. Purification (SiO2; 0-10% 2 M NH3 in MeOH/CH2Cl2) afforded the title compound as a clear oil (65 mg, 76%). HPLC: Rt=5.90. MS (ESI): mass calcd. for C30H34ClN5O4S, 596.2; m/z found, 597.4 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.8, 1H), 7.72 (s, 1H), 7.57-7.51 (m, 4H), 7.48-7.42 (m, 2H), 4.52 (s, 2H), 4.09 (t, J=6.8, 2H), 3.69 (t, J=4.6, 4H), 3.64 (t, J=5.8, 2H), 2.90 (s, 3H), 2.90-2.86 (m, 2H), 2.42-2.38 (m, 4H), 2.32 (t, J=6.8, 2H), 2.18 (s, 3H), 2.10-2.04 (m, 2H).
To a pressure tube containing a mixture of 3-(4-chloro-3-phenylethynyl-phenyl)-5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine (100 mg, 0.185 mmol), and Lindlar's catalyst (100 mg) in EtOAc (6 mL) was added quinoline (75 mg). The tube was placed in a shaker under 15 psi of H2 for 5 h. The reaction mixture was filtered through diatomaceous earth and concentrated to give a yellow oil. Purification (SiO2; 0-3% 2 M NH3 in MeOH/CH2Cl2) provided the title compound as a clear oil. The compound was obtained and analyzed as a 1.5:1 mixture of atropisomers at rt. HPLC: Rt=5.11 (major), 5.24 (minor). MS (ESI): mass calcd. for C28H33ClN4O3S, 541.1; m/z found, 542.3 [M+H]+. Major atropisomer: 1H NMR (CDCl3): 7.81-7.15 (m, 4.8H), 6.73 (d, J=12.2, 0.6H), 6.68 (d, J=12.2, 0.6H), 4.04 (s, 1.2H), 4.02 (t, J=6.8, 1.2H), 3.67 (t, J=4.7, 2.4H), 3.53 (t, J=5.8, 1.2H), 2.79 (t, J=5.7, 1.2H), 2.77 (s, 1.8H), 2.39-2.34 (br m, 2.4H), 2.28 (t, J=6.9, 1.2H), 2.04-1.96 (m, 1.2H). Minor atropisomer: 1H NMR (CDCl3): 7.81-7.15 (m, 4H), 4.53 (s, 0.8H), 4.10 (t, J=6.8, 0.8H), 3.70 (t, J=4.5, 1.6H), 3.64 (t, J=5.7, 0.8H), 2.90 (s, 1.2H), 2.88 (t, J=4.4, 0.8H), 2.43-2.39 (br m, 1.6H), 2.33 (t, J=6.7, 0.8H), 2.11-2.04 (m, 0.8H).
A solution of 3-(4-chloro-3-iodo-phenyl)-5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine (100 mg, 0.177 mmol), trans-cinnamylboronic acid (29 mg, 0.195 mmol), PdCl2(dppf)-CH2Cl2 (14.5 mg, 0.018 mmol), and K3PO4 (45 mg, 0.212 mmol) in degassed DMF (1.0 mL) was heated at 50° C. under N2 for 16 h. After cooling to rt, the reaction mixture was diluted with H2O and extracted with CH2Cl2 (2×). The combined organic extracts were washed with brine, dried (Na2SO4), filtered, and concentrated to give a brown oil. Purification (SiO2; 0-3% 2 M NH3 in MeOH/CH2Cl2) provided the title compound as a clear oil (57 mg, 60%). HPLC: Rt=5.29. MS (ESI): mass calcd. for C28H33ClN4O3S, 541.1; m/z found, 542.3 [M+H]+. 1H NMR (CDCl3): 7.97 (d, J=2.0, 1H), 7.59-7.56 (m, 2H), 7.52 (d, J=16.3, 1H), 7.46-7.28 (m, 5H), 7.16 (d, J=16.3, 1H), 4.55 (s, 2H), 4.13 (t, J=6.9, 2H), 3.71 (t, J=4.6, 4H), 3.67 (t, J=5.8, 2H), 2.91 (t, J=5.8, 2H), 2.89 (s, 3H), 2.45-2.40 (br m, 4H), 2.35 (t, J=6.8, 2H), 2.13-2.05 (m, 2H).
To a solution of (4-{2-chloro-5-[5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-phenylethynyl}-benzyl)-carbamic acid tert-butyl ester (622 mg, 0.931 mmol) in CH2Cl2 (4.7 mL) was added TFA (1.2 mL). The reaction mixture was stirred at rt for 1 h and then concentrated. Purification (SiO2; 0-5% 2 M NH3 in MeOH/CH2Cl2) afforded the title compound as a white foam (400 mg, 76%). MS (ESI): mass calcd. for C29H34ClN5O3S, 568.1; m/z found, 569.5 [M+H]+.
To a solution of 4-{2-chloro-5-[5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-phenylethynyl}-benzylamine (70 mg, 0.123 mmol) and pyridine (22 μL, 0.272 mmol) in CH2Cl2 (1.2 mL) was added MsCl (11 μL, 0.136 mmol). The reaction mixture was stirred at rt for 16 h and transferred directly to a silica gel column. Purification (SiO2; 0-3% 2 M NH3 in MeOH/CH2Cl2) afforded the title compound as a clear oil (65 mg, 82%). HPLC: Rt=4.73. MS (ESI): mass calcd. for C30H36ClN5O5S2, 646.2; m/z found, 647.5 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=1.8, 1H), 7.61-7.57 (m, 2H), 7.51-7.44 (m, 2H), 7.38-7.34 (m, 2H), 4.98 (br t, J=6.0, 1H), 4.53 (s, 2H), 4.34 (d, J=6.1, 2H), 4.10 (t, J=6.8, 2H), 3.69 (t, J=4.5, 4H), 3.65 (t, J=5.8, 2H), 2.90 (s, 3H), 2.90-2.87 (m, 2H), 2.89 (s, 3H), 2.43-2.38 (m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.03 (m, 2H).
The compounds in Examples 71-72 were prepared using methods analogous to those described in Example 70, substituting the appropriate sulfonyl chloride or acid chloride for MsCl.
HPLC: Rt=5.19. MS (ESI): mass calcd. for C35H38ClN5O5S2, 708.3; m/z found, 709.5 [M+H]+. 1H NMR (CDCl3): 7.88-7.84 (m, 1H), 7.78 (d, J=1.9, 1H), 7.62-7.57 (m, 1H), 7.54-7.45 (m, 6H), 7.19 (d, J=8.2, 2H), 5.10 (t, J=6.1, 1H), 4.52 (s, 2H), 4.16 (d, J=6.2, 2H), 4.10 (t, J=6.8, 2H), 3.68 (t, J=4.5, 4H), 3.64 (t, J=5.8, 2H), 2.90 (s, 3H), 2.90-2.87 (m, 2H), 2.42-2.37 (m, 4H), 2.32 (t, J=6.8, 2H), 2.10-2.03 (m, 2H).
MS (ESI): mass calcd. for C31H36ClN5O4S, 610.2; m/z found, 611.5 [M+H]+.
HPLC: Rt=5.05. MS (ESI): mass calcd. for C36H38ClN5O4S, 672.3; m/z found, 673.5 [M+H]+. 1H NMR (CDCl3): 7.83-7.79 (m, 3H), 7.58-7.54 (m, 2H), 7.53-7.41 (m, 5H), 7.35 (d, J=8.2, 2H), 6.65 (br t, J=5.6, 1H), 4.66 (d, J=5.8, 2H), 4.52 (s, 2H), 4.09 (t, J=6.7, 2H), 3.68 (t, J=4.6, 4H), 3.64 (t, J=5.8, 2H), 2.89 (s, 3H), 2.89-2.86 (m, 2H), 2.42-2.37 (br m, 4H), 2.31 (t, J=6.8, 2H), 2.10-2.02 (m, 2H).
A solution of 4-{2-chloro-5-[5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-phenylethynyl}-benzylamine (87 mg, 0.153 mmol), benzaldehyde (17.2 μL, 0.17 mmol), and ACOH (9 μL, 0.153 mmol) in CH2Cl2 (1.5 mL) was stirred at rt for 30 min. The mixture was treated with NaB(OAc)3H (42 mg, 0.20 mmol) and was stirred for 16 h. The reaction mixture was transferred directly to a silica gel column and purified (SiO2; 0-5% 2 M NH3 in MeOH/CH2Cl2) to afford the title compound as a clear oil (25 mg, 25%). HPLC: Rt=4.37. MS (ESI): mass calcd. for C36H40ClN5O3S, 658.3; m/z found, 659.5 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=1.8, 1H), 7.58-7.55 (m, 2H), 7.50-7.45 (m, 2H), 7.37-7.33 (m, 6H), 7.29-7.25 (m, 1H), 4.53 (s, 2H), 4.10 (t, J=6.8, 2H), 3.83 (s, 2H), 3.81 (s, 2H), 3.69 (t, J=4.5, 4H), 3.65 (t, J=5.7, 2H), 2.90 (s, 3H), 2.89 (t, J=5.7, 2H), 2.42-2.38 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.03 (m, 2H), 1.73 (br s, 1H).
A solution of 4-{2-chloro-5-[5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-phenylethynyl}-benzaldehyde (50 mg, 0.088 mmol), 4-methylbenzylamine (13.4 μL, 0.106 mmol), and ACOH (6 μL, 0.106 mmol) in CH2Cl2 (1.0 mL) were stirred at rt for 30 min. The mixture was treated with NaB(OAc)3H (24 mg, 0.106 mmol) and was stirred for 14 h. The reaction mixture was transferred directly to a silica gel column and purified (SiO2; 0-5% 2 M NH3 in MeOH/CH2Cl2) to afford the title compound as a clear oil (30 mg, 50%). MS (ESI): mass calcd. for C37H42ClN5O3S, 672.3; m/z found, 673.2 [M+H]+.
The compounds in Examples 76-79 were prepared using methods analogous to those described in Example 75, substituting the appropriate amine for 4-methylbenzylamine.
MS (ESI): mass calcd. for C36H39Cl2N5O3S, 692.7; m/z found, 693.2 [M+H]+.
MS (ESI): mass calcd. for C37H42ClN5O3S, 672.3; m/z found, 673.2 [M+H]+.
MS (ESI): mass calcd. for C33H40ClN5O3S, 622.2; m/z found, 623.2 [M+H]+.
MS (ESI): mass calcd. for C34H42ClN5O3S, 636.3; m/z found, 637.3 [M+H]+.
A. 2-(3-[2-Chloro-5-[5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-phenyl}-prop-2-ynyl)-isoindole-1,3-dione. This compound was prepared according to the methods described for Intermediate 1, substituting N-propargylphthalimide for TMSA in Step E. HPLC: Rt=4.48. MS (ESI): mass calcd. for C31H32ClN5O5S, 622.2; m/z found, 623.4 [M+H]+. 1H NMR (CDCl3): 7.90 (dd, J=5.4, 3.0, 2H), 7.75 (dd, J=5.5, 3.1, 2H), 7.68 (d, J=2.1, 1H), 7.45 (dd, J=8.5, 2.1, 1H), 7.37 (d, J=8.4, 1H), 4.75 (s, 2H), 4.48 (s, 2H), 4.07 (t, J=6.8, 2H), 3.68 (t, J=4.5, 4H), 3.63 (t, J=5.7, 2H), 2.90 (s, 3H), 2.87 (t, J=5.7, 2H), 2.42-2.37 (m, 4H), 2.30 (t, J=6.8, 2H), 2.08-2.01 (m, 2H).
B. 3-[2-Chloro-5-[5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-phenyl]-prop-2-ynylamine. To a solution of the above phthalimide (172 mg, 0.276 mmol) in 1:1 EtOH/DMF (2 mL) was added hydrazine monohydrate (134 μL). The reaction mixture was stirred at rt for 16 h, diluted with satd. aq. NaHCO3, and extracted with CH2Cl2 (3×). The combined organic extracts were dried (Na2SO4), filtered, and concentrated to give a yellow oil. Purification (SiO2; 0-10% 2 M NH3 in MeOH/CH2Cl2) afforded the title compound as a white solid (86 mg, 63%). HPLC: Rt=3.84. MS (ESI): mass calcd. for C23H30ClN5O3S, 492.0; m/z found, 493.4 [M+H]+. 1H NMR (CDCl3): 7.67 (d, J=2.1, 1H), 7.49 (dd, J=8.4, 2.1, 1H), 7.41 (d, J=8.4, 1H), 4.52 (s, 2H), 4.09 (t, J=6.8, 2H), 3.73 (s, 2H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 2.90 (s, 3H), 2.89 (t, J=5.7, 2H), 2.43-2.38 (m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.04 (m, 2H), 1.58 (br s, 2H).
C. 3-[2-Chloro-5-[5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-phenyl]-propylamine. This compound was prepared from the above propynyl amine according to the methods described in Example 2. HPLC: Rt=3.97. MS (ESI): mass calcd. for C23H34ClN5O3S, 496.1; m/z found, 497.5 [M+H]+. 1H NMR (CDCl3): 7.54 (d, J=2.0, 1H), 7.36 (d, J=8.3, 1H), 7.30 (dd, J=8.3, 2.2, 1H), 4.50 (s, 2H), 4.09 (t, J=6.8, 2H), 3.70 (t, J=4.5, 4H), 3.65 (t, J=5.8, 2H), 3.28 (t, J=7.1, 2H), 2.90-2.80 (m, 4H), 2.88 (s, 3H), 2.43-2.38 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.10-1.99 (m, 6H).
D. N-(3-[2-Chloro-5-[5-methanesulfonyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl-phenyl]-propyl)-methanesulfonamide.
To a solution of the above amine (50 mg, 0.101 mmol) and pyridine (20 μL) in CH2Cl2 (1.0 mL) was added MsCl (10 μL, 0.121 mmol). The reaction mixture was stirred at rt for 16 h. The mixture was concentrated and the crude residue transferred directly to a silica gel column. Purification (SiO2; 0-3% 2 M NH3 in MeOH/CH2Cl2) provided the title compound as a white foam (49 mg, 84%). HPLC: Rt=4.42. MS (ESI): mass calcd. for C24H36ClN5O5S2, 574.2; m/z found, 575.4 [M+H]+. 1H NMR (CDCl3): 7.49 (s, 1H), 7.37 (d, J=8.3, 1H), 7.38-7.36 (m, 2H), 4.69 (t, J=6.1, 1H), 4.51 (s, 2H), 4.09 (t, J=6.9, 2H), 3.70 (t, J=4.6, 4H), 3.64 (t, J=5.8, 2H), 3.19 (q, J=6.7, 2H), 2.96 (s, 3H), 2.91 (s, 3H), 2.90-2.83 (m, 4H), 2.43-2.38 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.10-2.03 (m, 2H), 1.98-1.91 (m, 2H).
The compounds in Examples 81-87 were prepared using methods analogous to those described in Example 80, Step D, substituting the appropriate sulfonyl chloride, acid chloride, or sulfamoyl chloride for MsCl.
MS (ESI): mass calcd. for C30H40ClN5O5S2, 650.3; m/z found, 651.5 [M+H]+.
MS (ESI): mass calcd. for C29H36Cl3N5O5S2, 705.1; m/z found, 706.4 [M+H]+.
MS (ESI): mass calcd. for C29H36Cl3N5O5S2, 705.1; m/z found, 706.4 [M+H]+.
MS (ESI): mass calcd. for C30H40ClN5O5S2, 650.3; m/z found, 651.5 [M+H]+.
MS (ESI): mass calcd. for C30H40ClN5O6S2, 666.3; m/z found, 667.5 [M+H]+.
MS (ESI): mass calcd. for C25H39ClN6O5S2, 603.2; m/z found, 604.4 [M+H]+.
MS (ESI): mass calcd. for C25H36ClN5O4S, 538.1; m/z found, 539.5 [M+H]+.
A. 1-(1-{3-[3-(4-Chloro-3-iodo-phenyl)-5-methanesulfonyl-4,5,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-1-yl]-propyl}-piperidin-4-yl)-pyrrolidin-2-one. This compound was prepared according to the method described in Intermediate 1, Step D, substituting 1-piperidin-4-yl-pyrrolidin-2-one for morpholine. HPLC: Rt=4.65. MS (ESI): mass calcd. for C25H33CIIN5O3S, 646.0; m/z found, 647.3 [M+H]+. 1H NMR (CDCl3): 8.15 (s, 1H), 7.45 (m, 2H), 4.48 (s, 2H), 4.07 (t, J=6.8, 2H), 4.00-3.91 (m, 1H), 3.64 (t, J=5.7, 2H), 3.35 (t, J=7.0, 2H), 2.93-2.86 (m, 4H), 2.90 (s, 3H), 2.39 (t, J=8.1, 2H), 2.32 (t, J=6.9, 2H), 2.08-1.97 (m, 6H), 1.70-1.63 (m, 4H).
B. 2-[3-[2-Chloro-5-(5-methanesulfonyl-1f-3-[4-(2-oxo-pyrrolidin-1-yl)-piperidin-1-yl]-pro yl]-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-phenyl-prop-2-ynyl]-isoindole-1,3-dione. This compound was prepared from the above iodide according to the method described in Example 80, Step A. HPLC: Rt=4.79. MS (ESI): mass calcd. for C36H39ClN6O5S, 703.3; m/z found, 704.4 [M+H]+. 1H NMR (CDCl3): 7.90 (dd, J=5.4, 3.1, 2H), 7.75 (dd, J=5.4, 3.1, 2H), 7.68 (d, J=2.0, 1H), 7.45 (dd, J=8.4, 2.1, 1H), 7.37 (d, J=8.4), 4.75 (s, 2H), 4.48 (s, 2H), 4.06 (t, J=6.7, 2H), 4.00-3.91 (m, 1H), 3.64 (t, J=5.7, 2H), 3.35 (t, J=7.0, 2H), 2.93-2.85 (m, 4H), 2.91 (s, 3H), 2.38 (t, J=8.1, 2H), 2.32 (t, J=6.9, 2H), 2.08-1.96 (m, 6H), 1.70-1.62 (m, 4H).
C. 2-[3-[2-Chloro-5-(5-methanesulfonyl-1-[3-[4-(2-oxo-pyrrolidin-1-yl)-piperidin-1-yl]-propyl]-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-phenyl]-propyl]-isoindole-1,3-dione. This compound was prepared from the above alkyne according to the method described in Example 2. HPLC: Rt=4.82. MS (ESI): mass calcd. for C36H43ClN6O5S, 707.3; m/z found, 708.4 [M+H]+. 1H NMR (CDCl3): 7.84 (dd, J=5.4, 3.0, 2H), 7.72 (dd, J=5.4, 3.1, 2H), 7.57 (d, J=2.0, 1H), 7.35-7.33 (m, 2H), 4.54 (s, 2H), 4.08 (t, J=6.8, 2H), 4.00-3.91 (m, 1H), 3.79 (t, J=7.0, 2H), 3.65 (t, J=5.6, 2H), 3.35 (t, J=7.0, 2H), 2.93-2.86 (m, 4H), 2.92 (s, 3H), 2.83 (t, J=7.5, 2H), 2.38 (t, J=8.1, 2H), 2.33 (t, J=6.9, 2H), 2.10-1.97 (m, 8H), 1.70-1.62 (m, 4H).
D. 2-[3-[2-Chloro-5-(5-methanesulfonyl-1-{3-[4-(2-oxo-pyrrolidin-1-yl)-piperidin-1-yl]-propyl}-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-phenyl]1-propylsulfamoyl}-benzoic acid methyl ester. To a solution of the above phthalimide (461 mg, 0.656 mmol) in EtOH (3.3 mL) was added hydrazine monohydrate (319 μL, 6.56 mmol). The reaction mixture was stirred at rt for 18 h. The resulting precipitate was filtered, and the filter cake washed with CH2Cl2. The filtrate was concentrated to provide a white solid, which was used directly in the next reaction without further purification.
To a solution of the crude amine (74 mg, 0.128 mmol) and pyridine (46 μL, 0.563 mmol) in CH2Cl2 (1.3 mL) was added 2-chlorosulfonyl-benzoic acid methyl ester (66 mg, 0.282 mmol). The reaction mixture was stirred at rt for 26 h and then transferred directly to a silica gel column. Purification (SiO2; 0-5% 2 M NH3 in MeOH/CH2Cl2) afforded the title compound as a clear oil (88 mg, 89%). HPLC: Rt=4.85. MS (ESI): mass calcd. for C36H47ClN6O7S2, 775.4; m/z found, 776.3 [M+H]+. 1H NMR (CDCl3): 8.09-8.06 (m, 1H), 7.84-7.81 (m, 1H), 7.64-7.61 (m, 2H), 7.45 (s, 1H), 7.35-7.33 (m, 2H), 6.04 (t, J=6.1, 1H), 4.52 (s, 2H), 4.08 (t, J=6.7, 2H), 4.00-3.92 (m, 1H), 3.99 (s, 3H), 3.65 (t, J=5.6, 2H), 3.35 (t, J=7.0, 2H), 3.06 (q, J=6.5, 2H), 2.93-2.86 (m, 4H), 2.91 (s, 3H), 2.79 (t, J=7.5, 2H), 2.38 (t, J=8.1, 2H), 2.33 (t, J=6.9, 2H), 2.09-1.96 (m, 6H), 1.88-1.80 (m, 2H), 1.70-1.62 (m, 4H).
E. 2-{3-[2-Chloro-5-(5-methanesulfonyl-1-{3-[4-(2-oxo-pyrrolidin-1-yl)-piperidin-1-yl]-propyl}-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-phenyl]-propylsulfamoyl}-benzoic acid, trifluoroacetic acid salt. To a solution of the above ester (63 mg, 0.078 mmol) in THF (0.8 mL) was added LiOH (2 M in H2O; 0.2 mL). The reaction mixture was stirred at rt for 12 h, and then purified directly by reverse-phase HPLC (0.05% TFA/10-98% MeCN/H2O) to provide the TFA salt of the title compound as a white solid (63 mg, 91%). HPLC: Rt=4.68. MS (ESI): mass calcd. for C35H45ClN6O7S2, 761.4; m/z found, 762.2 [M+H]+. 1H NMR (CD3OD): 7.98-7.94 (m, 1H), 7.83-7.78 (m, 1H), 7.70-7.61 (m, 2H), 7.58 (s, 1H), 7.43-7.39 (m, 2H), 4.46 (s, 2H), 4.26 (t, J=6.0, 2H), 4.10-4.02 (m, 1H), 3.67-3.58 (br m, 2H), 3.35-3.24 (m, 4H), 3.17-2.95 (m, 4H), 2.97 (s, 3H), 2.92 (t, J=5.6, 2H), 2.81 (t, J=7.3, 2H), 2.34 (t, J=8.0, 2H), 2.33-2.29 (m, 2H), 2.01-1.94 (m, 4H), 1.90-1.78 (m, 6H).
The compounds in Examples 89-100 were prepared using methods analogous to those described in Example 88, Steps A-D, with the appropriate substituent changes.
HPLC: Rt=4.82. MS (ESI): mass calcd. for C34H44ClN7O7S2, 762.4; m/z found, 763.3 [M+H]+. 1H NMR (CDCl3): 8.13-8.10 (m, 1H), 7.85-7.82 (m, 1H), 7.74-7.71 (m, 2H), 7.45 (d, J=1.5, 1H), 7.35-7.33 (m, 2H), 5.54 (br s, 1H), 4.50 (s, 2H), 4.08 (t, J=6.8, 2H), 4.00-3.91 (m, 1H), 3.65 (t, J=5.7, 2H), 3.35 (t, J=7.0, 2H), 3.17 (t, J=6.4, 2H), 2.93-2.86 (m, 4H), 2.91 (s, 3H), 2.80 (t, J=7.5, 2H), 2.38 (t, J=8.1, 2H), 2.33 (t, J=6.9, 2H), 2.10-1.96 (m, 6H), 1.93-1.85 (m, 2H), 1.70-1.62 (m, 4H).
HPLC: Rt=4.95. MS (ESI): mass calcd. for C34H44ClN6O5S2, 751.8; m/z found, 753.3 [M+H]+. 1H NMR (CDCl3): 7.85 (t, J=1.8, 1H), 7.76-7.73 (m, 1H), 7.55-7.52 (m, 1H), 7.47-7.42 (m, 2H), 7.35-7.33 (m, 2H), 5.19 (br t, J=5.7, 1H), 4.51 (s, 2H), 4.07 (t, J=6.8, 2H), 4.00-3.91 (m, 1H), 3.65 (t, J=5.7, 2H), 3.35 (t, J=7.0, 2H), 3.17 (t, J=6.4, 2H), 3.00 (dd, J=12.8, 6.6, 4H), 2.94-2.86 (m, 4H), 2.91 (s, 3H), 2.79 (t, J=7.5, 2H), 2.39 (t, J=8.1, 2H), 2.34 (t, J=6.9, 2H), 2.10-1.96 (m, 6H), 1.89-1.85 (m, 2H), 1.70-1.62 (m, 4H).
MS (ESI): mass calcd. for C34H45ClN6O5S2, 717.4; m/z found, 718.5 [M+H]+.
MS (ESI): mass calcd. for C35H47ClN6O5S2, 731.4; m/z found, 732.5 [M+H]+.
MS (ESI): mass calcd. for C35H47ClN6O6S2, 747.4; m/z found, 748.3 [M+H]+.
MS (ESI): mass calcd. for C35H47ClN6O5S2, 731.4; m/z found, 732.3 [M+H]+.
MS (ESI): mass calcd. for C34H44Cl2N6O5S2, 751.8; m/z found, 753.2 [M+H]+.
MS (ESI): mass calcd. for C34H44ClN7O7S2, 762.4; m/z found, 763.2 [M+H]+.
MS (ESI): mass calcd. for C35H47ClN6O5S2, 731.4; m/z found, 732.3 [M+H]+.
MS (ESI): mass calcd. for C35H44ClN7O5S2, 742.4; m/z found, 743.3 [M+H]+.
MS (ESI): mass calcd. for C35H47ClN6O7S3, 795.5; m/z found, 796.2 [M+H]+.
MS (ESI): mass calcd. for C35H47ClN6O7S3, 795.5; m/z found, 796.3 [M+H]+.
A. 1-[1-(3-[3-{4-Chloro-3-(3-hydroxy-prop-1-ynyl)-phenyl]-5-methanesulfonyl-4,5,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-1-yl}-propyl)-piperidin-4-yl]-pyrrolidin-2-one. This compound was prepared following the methods described in Example 80, Step A, substituting propargyl alcohol for N-propargylphthalimide. HPLC Rt=4.30. MS (ESI): mass calcd. for C28H36ClN5O4S, 574.2; m/z found, 575.3 [M+H]+. 1H NMR (CDCl3): 7.64 (d, J=2.1, 1H), 7.48 (dd, J=8.4, 2.1, 1H), 7.39 (d, J=8.4, 1H), 4.54 (s, 2H), 4.46 (s, 2H), 4.05 (t, J=6.8, 2H), 3.99-3.91 (m, 1H), 3.61 (t, J=5.7, 2H), 3.35 (t, J=7.0, 2H), 2.94 (br s, 1H), 2.91 (s, 3H), 2.89-2.83 (m, 4H), 2.39 (t, J=7.9, 2H), 2.34 (t, J=7.2, 2H), 2.10-1.97 (m, 6H), 1.71-1.64 (m, 4H).
B. 1-fi-(3-[3-{4-Chloro-3-(3-hydroxy-propyl)-phenyl]-5-methanesulfonyl-4,5,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-4-yl-pyrrolidin-2-one. This compound was prepared from the above alkyne following the methods described in Example 2. MS (ESI): mass calcd. for C28H40ClN5O4S, 578.2; m/z found, 579.3.
C. 1-fi-(3-[3-[4-Chloro-3-(3-pyrrolidin-1-yl-propyl)-phenyl]-5-methanesulfonyl-4,5,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-1-yl]-propyl)-piperidin-4-yl]-pyrrolidin-2-one. To a solution of the above alcohol (584 mg, 1.01 mmol) and pyridine (250 μL, 3.03 mmol) in CH2Cl2 (8 mL) was added MsCl (120 μL, 1.52 mmol). The reaction mixture was stirred at rt for 4 h, diluted with H2O, and extracted with CH2Cl2 (3×). The combined organic extracts were dried (Na2SO4), filtered, and concentrated to give a tan solid, which was used directly without further purification. To a solution of the crude mesylate (100 mg, 0.152 mmol) in EtOH (1.0 mL) was added pyrrolidine (3.04 mmol). The reaction mixture was heated to 60° C. and stirred for 16 h. The solvent was removed and the crude residue was purified (SiO2; 2 M NH3 in MeOH/CH2Cl2) to provide the title compound as a white solid (54 mg, 71%). HPLC: Rt=4.01. MS (ESI): mass calcd. for C32H47ClN6O3S, 631.3; m/z found, 632.3 [M+H]+. 1H NMR (CDCl3): 7.52 (d, J=2.0, 1H), 7.36 (d, J=8.2, 1H), 7.29 (dd, J=8.3, 2.1, 1H), 4.50 (s, 2H), 4.08 (t, J=6.8, 2H), 4.01-3.93 (m, 1H), 3.65 (t, J=5.7, 2H), 3.35 (t, J=7.0, 2H), 2.93-2.86 (m, 4H), 2.89 (s, 3H), 2.83-2.78 (m, 2H), 2.55-2.49 (m, 6H), 2.39 (t, J=7.9, 2H), 2.33 (t, J=7.0, 2H), 2.09-2.03 (m, 8H), 2.09-1.97 (m, 6H), 1.92-1.85 (m, 2H), 1.80-1.76 (m, 4H), 1.70-1.63 (m, 4H).
The compounds in Examples 102-108 were prepared using methods analogous to those described in Example 101 with the appropriate substituent changes.
MS (ESI): mass calcd. for C33H49ClN6O3S, 645.3; m/z found, 646.3 [M+H]+.
MS (ESI): mass calcd. for C34H51ClN6O3S, 659.3; m/z found, 660.4 [M+H]+.
MS (ESI): mass calcd. for C33H47ClF2N6O3S, 681.3; m/z found, 682.4 [M+H]+.
MS (ESI): mass calcd. for C32H47ClN6O4S, 647.3; m/z found, 648.3 [M+H]+.
MS (ESI): mass calcd. for C33H50ClN7O3S, 660.3; m/z found, 661.4 [M+H]+.
MS (ESI): mass calcd. for C34H51ClN6O3S, 659.3; m/z found, 660.3 [M+H]+.
MS (ESI): mass calcd. for C33H49ClN6O3S, 645.3; m/z found, 646.3 [M+H]+.
A. 1-[1-(3-[3-{4-Chloro-3-(4-hydroxy-but-1-ynyl)-phenyl]-5-methanesulfonyl-4,5,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-1-yl}-prolyl)-piperidin-4-yl]-pyrrolidin-2-one. This compound was prepared following the methods described in Example 101, Step A, substituting for 3-butyn-1-ol for propargyl alcohol. HPLC: Rt=4.33. MS (ESI): mass calcd. for C29H38ClN5O4S, 588.2; m/z found, 589.3 [M+H]+. 1H NMR (CDCl3): 7.68 (d, J=1.3, 1H), 7.45 (dd, J=8.4, 1.6, 1H), 7.40 (d, J=8.4, 1H), 4.49 (s, 2H), 4.06 (t, J=6.6, 2H), 3.99-3.91 (m, 1H), 3.85 (t, J=6.2, 1H), 3.63 (t, J=5.7, 2H), 3.35 (t, J=7.0, 2H), 2.93-2.84 (m, 4H), 2.90 (s, 3H), 2.76 (t, J=6.3, 2H), 2.39 (t, J=7.9, 2H), 2.33 (t, J=7.2, 2H), 2.09-1.97 (m, 6H), 1.70-1.62 (m, 4H).
B. 1-[1-(3-{3-[4-Chloro-3-(4-hydroxy-butyl)-phenyl]-5-methanesulfonyl-4,5,6,7-tetrahydro-pyrazolo[43-c]pyridin-4-yl-pyrrolidin-2-one. This compound was prepared from the above alkyne following the methods described in Example 101, Step B. MS (ESI): mass calcd. for C29H42ClN5O4S, 592.2; m/z found, 593.4 [M+H]
C. 1-[1-(3-{3-[4-Chloro-3-(4-pyrrolidin-1-yl-butyl)-phenyl]-5-methanesulfonyl-4,5,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-1-yl}-propyl)-piperidin-4-yl]-pyrrolidin-2-one. The title compound was prepared from the above alcohol following the methods described in Example 101, Step C. HPLC: Rt=4.07. MS (ESI): mass calcd. for C33H49ClN6O3S, 645.3; m/z found, 646.4 [M+H]+. 1H NMR (CDCl3): 7.50 (d, J=2.0, 1H), 7.35 (d, J=8.2, 1H), 7.28 (dd, J=8.3, 2.1, 1H), 4.50 (s, 2H), 4.08 (t, J=6.8, 2H), 4.01-3.92 (m, 1H), 3.65 (t, J=5.7, 2H), 3.35 (t, J=7.0, 2H), 2.92-2.87 (m, 4H), 2.89 (s, 3H), 2.78 (t, J=7.3, 2H), 2.54-2.47 (m, 6H), 2.39 (t, J=7.9, 2H), 2.33 (t, J=7.0, 2H), 2.09-1.98 (m, 6H), 1.80-1.75 (m, 4H), 1.71-1.60 (m, 8H).
The compounds in Examples 110-115 were prepared using methods analogous to those described in Example 109 with the appropriate substituent changes.
MS (ESI): mass calcd. for C34H51ClN6O3S, 659.3; m/z found, 660.4 [M+H]+.
MS (ESI): mass calcd. for C35H53ClN6O3S, 673.4; m/z found, 674.4 [M+H]+.
MS (ESI): mass calcd. for C34H52ClN7O3S, 674.4; m/z found, 675.4 [M+H]+.
MS (ESI): mass calcd. for C33H49ClN6O4S, 661.3; m/z found, 662.4 [M+H]+.
MS (ESI): mass calcd. for C35H53ClN6O3S, 673.4; m/z found, 674.5 [M+H]+.
MS (ESI): mass calcd. for C34H51ClN6O3S, 659.3; m/z found, 660.4 [M+H]+.
A. 3-Amino-4-trifluoromethyl-benzoic acid. A mixture of 3-nitro-4-trifluoromethylbenzoic acid (19.5 g, 83 mmol) and 10% Pd/C (4 g) in EtOAc (300 mL) was shaken under 40 psi of H2 in a pressure vessel. The vessel was repeatedly refilled until the pressure stabilized at 40 psi. Shaking was continued for 10 min, and then N2 was bubbled through the mixture for 30 min. The resulting black mixture was filtered through diatomaceous earth and washed with MeOH. The filtrate was concentrated to afford the desired product as a white solid (19.5 g, 98%), which needed no further purification. HPLC: Rt=5.46. MS (ESI): mass calcd. for C8H6F3NO2, 205.1; m/z found, 204.3 [M−H]−. 1H NMR (CD3OD): 7.51 (s, 1H), 7.45 (d, J=8.2, 1H), 7.30 (d, J=8.0, 1H), 5.02 (br s, 3H).
B. 3-Iodo-4-trifluoromethyl-benzoic acid. To a −5 LC slurry of the above aniline (16.6 g, 80.9 mmol) and conc. H2SO4 (33 mL) in H2O (17 mL) was added a solution of NaNO2 (6.7 g, 97.1 mmol) in H2O (25 mL). The mixture was stirred at 0 LC for 1 h, then treated with a solution of KI (26.9 g, 161.8 mmol) and 12 (1 crystal) in H2O (30 mL). The reaction mixture was stirred for 1 h at rt and then heated at reflux for 4 h, during which time significant foaming and frothing was observed. The thick brown mixture was cooled to 0° C., diluted with satd. aq. Na2S2O3 (100 mL), and filtered to provide a beige solid. The solid was washed with H2O and recrystallized from CH3CN to afford the desired product as a white solid (18.3 g, 72%). HPLC: Rt=6.33. MS (ESI): mass calcd. for C8H4F3IO2, 316.0; m/z found, 315.2 [M−H]−. 1H NMR (CD3OD): 8.68 (s, 1H), 8.11 (d, J=7.9, 1H), 7.73 (d, J=8.2, 1H), 4.44 (br s, 1H).
C. 3-(3-Iodo-4-trifluoromethyl-phenyl)-5-methanesulfonyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine. To a 0 LC solution of the above acid (6.3 g, 20 mmol) and oxalyl chloride (1.9 mL, 22 mmol) in benzene (20 mL) was added DMF (0.15 mL). The reaction mixture was warmed to rt and stirred for 5 h. The mixture was concentrated to afford a white solid, which was used directly in the next reaction following the procedure outlined in Intermediate 1, Step B. HPLC: Rt=6.25. MS (ESI): mass calcd. for C14H13F31N3O2S, 471.2; m/z found, 472.3 [M+H]+. 1H NMR (DMSO-d6): 8.39 (s, 1H), 7.82 (d, J=8.3, 1H), 7.72 (d, J=8.3, 1H), 4.49 (s, 2H), 3.52 (t, J=5.7, 2H), 3.00 (s, 3H), 2.85 (t, J=5.6, 2H).
D. 1-[1-(3-[3-[3-(4-Chloro-phenylethynyl)-4-trifluoromethyl-phenyl]-5-methanesulfonyl-4,5,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-1-yl]-2-hydroxy-propyl)-piperidin-4-yl]-pyrrolidin-2-one. To a slurry of the above pyrazole (1.28 g, 2.72 mmol) and Cs2CO3 (1.33 g, 4.08 mmol) in DMF (9 mL) was added epichlorohydrin (1.26 g, 13.6 mmol). The reaction mixture was stirred at rt for 17 h, then cooled to 0 LC, and treated slowly with ice water. The resulting precipitate was filtered to provide the desired epoxide as a white solid, which was used directly in the next reaction. The crude epoxide (105 mg, 0.199 mmol) suspended in EtOH (1.0 mL) and treated with 1-piperidin-4-yl-pyrrolidin-2-one (37 mg, 0.219 mmol). The reaction mixture was heated at reflux for 16 h and then concentrated. Purification (SiO2; 0-3% 2 M NH3 in MeOH/CH2Cl2) provided a white solid, which was converted to the title compound following the procedure described in Intermediate 1, Step E, substituting 4-chloro-ethynylbenzene for TMSA. HPLC: Rt=5.45. MS (ESI): mass calcd. for C34H37ClF3N5O4S, 704.2; m/z found, 705.4 [M+H]+. 1H NMR (CDCl3): 7.91 (s, 1H), 7.72 (d, 8.3, 1H), 7.61 (d, J=8.3, 1H), 7.50 (d, J=8.5, 2H), 7.35 (d, J=8.6, 2H), 4.58 (dd, J=18.4, 14.6, 2H), 4.19 (dd, J=13.8, 2.8, 1H), 4.15-4.09 (br m, 1H), 4.02-3.94 (br m, 2H), 3.73-3.62 (m, 2H), 3.34 (t, J=7.0, 2H), 3.09-2.99 (m, 2H), 2.95-2.92 (m, 2H), 2.89 (s, 3H), 2.49-2.34 (m, 5H), 2.16-2.09 (m, 1H), 2.04-1.96 (m, 2H), 1.78-1.61 (m, 4H).
The compounds in Examples 117-122 were prepared using methods analogous to those described in Example 116, with the appropriate substituent changes.
HPLC: Rt=5.73. MS (ESI): mass calcd. for C32H34ClF3N4O5S, 679.2; m/z found, 680.4 [M+H]+. 1H NMR (CDCl3): 7.91 (s, 1H), 7.71 (d, J=8.3, 1H), 7.61 (d, J=8.2, 1H), 7.50 (d, J=8.6, 2H), 7.35 (d, J=8.6, 2H), 4.58 (dd, J=22.5, 14.4, 2H), 4.20 (dd, J=13.7, 2.9, 1H), 4.17-4.14 (m, 1H), 4.01 (dd, J=13.6, 6.5, 1H), 3.75-3.61 (m, 2H), 3.68 (s, 3H), 3.11-3.04 (m, 1H), 2.96-2.93 (m, 2H), 2.89 (s, 3H), 2.49-2.31 (m, 4H), 2.15-2.10 (m, 1H), 1.94-1.90 (m, 2H), 1.83-1.66 (m, 4H).
HPLC: Rt=5.43. MS (ESI): mass calcd. for C33H35ClF3N5O4S, 690.2; m/z found, 691.4 [M+H]+. 1H NMR (CDCl3): 7.91 (s, 1H), 7.71 (d, J=8.3, 1H), 7.60 (d, J=8.3, 1H), 7.50 (d, J=8.6, 2H), 7.35 (d, J=8.6, 2H), 6.37 (s, 1H), 4.58 (dd, J=21.4, 14.5, 2H), 4.20 (dd, J=13.7, 2.8, 1H), 4.16-4.12 (br m, 1H), 4.02 (dd, J=13.6, 6.6, 1H), 3.75-3.61 (m, 2H), 3.32 (t, J=6.9, 2H), 3.11-3.04 (m, 1H), 2.96-2.93 (m, 2H), 2.89 (s, 3H), 2.83-2.80 (br m, 1H), 2.50-2.37 (m, 3H), 2.18-2.12 (m, 1H), 2.02 (t, J=6.9, 2H), 2.01-1.85 (m, 3H), 1.46 (br m, 2H).
HPLC: Rt=5.31. MS (ESI): mass calcd. for C31H33ClF3N5O4S, 664.2; m/z found, 665.4 [M+H]+. 1H NMR (CDCl3): 7.91 (s, 1H), 7.71 (d, J=8.3, 1H), 7.60 (d, J=8.3, 1H), 7.50 (d, J=8.5, 2H), 7.35 (d, J=8.6, 2H), 6.58 (br s, 1H), 6.49 (br s, 1H), 4.57 (dd, J=19.1, 14.5, 2H), 4.19 (dd, J=13.8, 2.8, 1H), 4.16-4.09 (br m, 1H), 4.00 (dd, J=13.8, 6.7, 1H), 3.73-3.61 (m, 2H), 3.14-3.03 (m, 1H), 2.99-2.84 (m, 2H), 2.89 (s, 3H), 2.45-2.35 (m, 2H), 2.32-2.24 (m, 1H), 2.20-2.12 (m, 1H), 2.06-1.98 (m, 1H), 1.86-1.63 (m, 4H).
HPLC: Rt=5.69. MS (ESI): mass calcd. for C39H42ClF3N8O4S, 811.3; m/z found, 812.5 [M+H]+. 1H NMR (CDCl3): 7.92 (s, 1H), 7.71 (d, J=8.3, 1H), 7.62 (d, J=7.7, 1H), 7.50 (dt, J=2.1, 8.6, 2H), 7.35 (dt, J=2.1, 8.6, 2H), 7.03 (d, J=8.5, 1H), 6.16 (d, J=8.5, 1H), 4.59 (dd, J=23.5, 15.0, 2H), 4.41-4.33 (m, 1H), 4.23 (dd, J=13.8, 2.7, 1H), 4.19-4.13 (br m, 1H), 4.05 (dd, J=13.8, 6.8, 1H), 3.77-3.63 (m, 2H), 3.34 (s, 3H), 3.16-3.05 (m, 1H), 3.03 (s, 6H), 2.98-2.93 (m, 2H), 2.90 (s, 3H), 2.86-2.70 (m, 2H), 2.57-2.44 (m, 3H), 2.26-2.20 (m, 1H), 1.77-1.74 (br m, 2H).
HPLC: Rt=5.96. MS (ESI): mass calcd. for C35H41ClF3N5O5S, 736.3; m/z found, 737.5 [M+H]+. 1H NMR (CDCl3): 7.91 (s, 1H), 7.71 (d, J=8.3, 1H), 7.60 (d, J=8.8, 1H), 7.52-7.48 (m, 2H), 7.38-7.33 (m, 2H), 4.57 (dd, J=20.5, 14.5, 2H), 4.47 (br s, 1H), 4.18 (dd, J=13.8, 2.8, 1H), 4.15-4.07 (br m, 1H), 3.99 (dd, J=13.8, 6.8, 1H), 3.73-3.60 (m, 2H), 3.47 (br s, 1H), 3.09-3.02 (m, 1H), 2.96-2.86 (m, 3H), 2.88 (s, 3H), 2.77-2.70 (br m, 1H), 2.47-2.34 (m, 3H), 2.12 (t, J=11.1, 1H), 1.93 (br d, J=11.1, 2H), 1.49-1.36 (m, 2H), 1.44 (s, 9H).
MS (ESI): mass calcd. for C29H30ClF3N4O4S, 623.1; m/z found, 624.4 [M+H]+.
The compounds in Examples 123-131 were prepared from 3-(4-chloro-3-iodo-phenyl)-5-methanesulfonyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine (Intermediate 1, Step B) according to the methods described in Example 116, Step D, substituting the appropriate amine for 1-piperidin-4-yl-pyrrolidin-2-one.
HPLC: Rt=5.29. MS (ESI): mass calcd. for C33H37Cl2N5O4S, 670.7; m/z found, 671.5 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=1.7, 1H), 7.54-7.47 (m, 4H), 7.35 (dd, J=6.6, 1.9, 2H), 4.55 (d, J=4.3, 2H), 4.17 (dd, J=13.8, 2.8, 1H), 4.13-4.07 (m, 1H), 4.02-3.96 (m, 2H), 3.73-3.62 (m, 3H), 3.34 (t, J=7.0, 2H), 3.08-2.99 (m, 2H), 2.94-2.91 (m, 3H), 2.85 (s, 3H), 2.45-2.37 (m, 5H), 2.00 (t, J=7.6, 2H), 1.75-1.61 (m, 4H).
HPLC: Rt=5.47. MS (ESI): mass calcd. for C28H30Cl2N4O4S, 589.6; m/z found, 590.2 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.5, 1H), 7.54-7.50 (m, 2H), 7.48-7.45 (m, 2H), 7.37-7.32 (m, 2H), 4.53 (dd, J=18.3, 14.5, 2H), 4.18 (dd, J=13.3, 2.9, 1H), 4.17-4.10 (m, 1H), 4.02-3.96 (m, 1H), 3.74-3.59 (m, 7H), 3.07-2.99 (m, 1H), 2.94-2.88 (m, 1H), 2.87 (s, 3H), 2.65-2.58 (m, 2H), 2.48-2.38 (m, 4H).
HPLC: Rt=5.64. MS (ESI): mass calcd. for C29H32Cl2N4O3S, 587.6; m/z found, 588.5 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=1.9, 1H), 7.53 (d, J=8.5, 2H), 7.49-7.47 (m, 2H), 7.35 (d, J=8.5, 2H), 4.55 (dd, J=24.4, 14.5, 2H), 4.16 (dd, J=13.7, 2.8, 1H), 4.13-4.08 (m, 1H), 3.98 (dd, J=13.7, 6.8, 1H), 3.73-3.60 (m, 2H), 3.12-3.05 (m, 1H), 2.96-2.90 (m, 1H), 2.88 (s, 3H), 2.56 (br m, 2H), 2.42-2.27 (m, 4H), 1.59-1.52 (m, 4H), 1.47-1.42 (m, 3H).
HPLC: Rt=5.60. MS (ESI): mass calcd. for C38H42Cl2N8O4S, 777.8; m/z found, 778.4 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=1.9, 1H), 7.53-7.45 (m, 4H), 7.34 (dd, J=6.6, 2.0, 2H), 7.03 (d, J=8.5, 1H), 6.16 (d, J=8.5, 1H), 4.56 (dd, J=23.7, 14.5, 2H), 4.40-4.32 (m, 1H), 4.21 (dd, J=13.8, 2.7, 1H), 4.17-4.11 (m, 1H), 4.02 (dd, J=13.8, 6.8, 1H), 3.76-3.61 (m, 2H), 3.33 (s, 3H), 3.16-3.07 (m, 2H), 3.03 (s, 6H), 2.96-2.90 (m, 2H), 2.89 (s, 3H), 2.86-2.69 (m, 2H), 2.53-2.42 (m, 3H), 2.23-2.18 (m, 1H), 1.78-1.71 (m, 2H).
HPLC: Rt=5.36. MS (ESI): mass calcd. for C31H34Cl2N4O5S, 645.6; m/z found, 646.4 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=1.8, 1H), 7.54-7.45 (m, 4H), 7.35 (dt, J=2.1, 8.6, 2H), 4.55 (dd, J=22.8, 14.5, 2H), 4.17 (dd, J=13.8, 2.8, 1H), 4.14-4.08 (m, 1H), 3.98 (dd, J=13.8, 6.8, 1H), 3.74-3.60 (m, 2H), 3.68 (s, 3H), 3.06 (dt, J=5.5, 16.1, 1H), 2.94-2.87 (m, 2H), 2.88 (s, 3H), 2.79-2.76 (br m, 1H), 2.44-2.29 (m, 4H), 2.08-2.03 (br m, 1H), 1.92-1.88 (br m, 3H), 1.80-1.63 (m, 2H).
HPLC: Rt=4.97. MS (ESI): mass calcd. for C30H33Cl2N5O4S, 630.6; m/z found, 631.3 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.6, 1H), 7.54-7.51 (m, 2H), 7.48-7.47 (m, 2H), 7.37-7.33 (m, 2H), 5.63 (br s, 1H), 5.53 (br s, 1H), 4.54 (dd, J=20.0, 14.5, 2H), 4.17 (dd, J=13.7, 2.8, 1H), 4.14-4.08 (m, 1H), 3.98 (dd, J=13.7, 6.6, 1H), 3.73-3.59 (m, 2H), 3.09-2.84 (m, 4H), 2.88 (s, 3H), 2.44-2.26 (m, 4H), 2.20-2.11 (m, 1H), 2.06-1.99 (m, 1H), 1.89-1.63 (m, 4H).
HPLC: Rt=5.53. MS (ESI): mass calcd. for C28H30Cl2N4O3S, 573.6; m/z found, 574.3 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=1.9, 1H), 7.54-7.51 (m, 2H), 7.50-7.46 (m, 2H), 7.36-7.33 (m, 2H), 4.54 (dd, J=20.2, 14.5, 2H), 4.18 (dd, J=13.8, 2.8, 1H), 4.13-4.07 (br m, 2H), 3.99 (dd, J=13.8, 7.0, 1H), 3.72-3.58 (m, 2H), 3.08-3.01 (m, 1H), 2.94-2.85 (m, 1H), 2.87 (s, 3H), 2.66-2.61 (m, 3H), 2.53-2.47 (m, 2H), 2.43 (dd, J=12.0, 4.4, 1H), 1.80-1.73 (m, 6H).
HPLC: Rt=5.51. MS (ESI): mass calcd. for C34H41Cl2N5O5S, 702.7; m/z found, 703.5 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.6, 1H), 7.52 (d, J=8.6, 2H), 7.48-7.46 (m, 2H), 7.35 (d, J=8.6, 2H), 4.54 (dd, J=20.6, 14.5, 2H), 4.48-4.45 (br s, 1H), 4.16 (dd, J=13.8, 2.7, 1H), 4.12-4.07 (br m, 1H), 3.97 (dd, J=13.8, 6.8, 1H), 3.72-3.59 (m, 2H), 3.46 (br s, 1H), 3.07-3.01 (m, 1H), 2.92-2.84 (m, 2H), 2.87 (s, 3H), 2.76-2.70 (br m, 1H), 2.44-2.34 (m, 3H), 2.11 (t, J=11.1, 1H), 1.96-1.88 (br m, 2H), 1.44 (s, 9H), 1.41-1.31 (m, 3H).
HPLC: Rt=5.57. MS (ESI): mass calcd. for C33H39Cl2N5O5S, 688.7; m/z found, 689.4 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.5, 1H), 7.55-7.51 (m, 2H), 7.49-7.47 (m, 2H), 7.37-7.34 (m, 2H), 4.55 (dd, J=19.9, 14.5, 2H), 4.22-4.09 (m, 3H), 4.00 (dd, J=13.6, 6.5, 1H), 3.74-3.61 (m, 2H), 3.47-3.38 (br m, 4H), 3.08-3.01 (m, 1H), 2.95-2.87 (m, 1H), 2.89 (s, 3H), 2.61-2.54 (m, 2H), 2.49-2.35 (m, 4H), 1.45 (s, 9H).
To a solution of 4-(3-{3-[4-chloro-3-(4-chloro-phenylethynyl)-phenyl]-5-methanesulfonyl-4,5,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-1-yl}-2-hydroxy-propyl)-piperazine-1-carboxylic acid tert-butyl ester (117 mg, 0.161 mmol) in CH2Cl2 (1.1 mL) was added TFA (0.5 mL). The reaction mixture was stirred at rt for 1 h and then concentrated to give a brown oil. Purification (SiO2; 0-10% 2 M NH3 in MeOH/CH2Cl2) afforded the title compound as a clear oil (80 mg, 84%). MS (ESI): mass calcd. for C28H31Cl2N5O3S, 588.6; m/z found, 589.3 [M+H]+.
This compound was prepared from [1-(3-{3-[4-chloro-3-(4-chloro-phenylethynyl)-phenyl]-5-methanesulfonyl-4,5,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-1-yl}-2-hydroxy-propyl)-piperidin-4-yl]-carbamic acid tert-butyl ester according to the method described in Example 132. MS (ESI): mass calcd. for C29H33Cl2N5O3S, 602.6; m/z found, 603.4 [M+H]+.
A. 7-Bromo-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester. To a solution of 7-bromo-1,2,3,4-tetrahydroisoquinoline hydrochloride (4.0 g, 16.1 mmol) and Et3N (6.7 mL, 48.3 mmol) in CH2Cl2 (500 mL) was added di-tert-butyl-dicarbonate (4.2 g, 19.2 mmol). After 18 h at rt, the mixture was concentrated and the product was purified on SiO2 (EtOAc/hexanes) to afford a clear oil (5.00 g, 99%).
B. 7-Trimethylsilanylethynyl-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester. To a solution of the above bromide (3.0 g, 9.6 mmol), PdCl2(PPh)3 (675 mg, 0.96 mmol), and CuI (183 mg, 0.96 mmol) in degassed DMF (50 mL) was added ethynyltrimethylsilane (2.7 mL, 19.2 mmol), followed by Et3N (4.0 mL, 28.9 mmol). The reaction mixture was stirred under N2 at 60° C. for 1 h. The mixture was diluted with satd. aq. NaHCO3 and extracted with EtOAc (×2). The combined organic extracts were washed with water (×3), dried (Na2SO4), filtered, and concentrated to give a brown oil. Purification on SiO2 (EtOAc/hexanes) afforded the desired product (2.78 g, 88%).
C. 7-Ethynyl-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester. To a solution of the above alkyne (2.7 g, 8.45 mmol) in THF (150 mL) was added tetrabutylammonium fluoride (1.0 M in THF; 16.9 mL, 16.9 mmol). After 1 h at rt, the mixture was diluted with water and extracted with EtOAc (×3). The combined organic extracts were washed with water (×3), dried (Na2SO4), filtered, and concentrated to give a brown oil. Purification on SiO2 (EtOAc/hexanes) provided the title compound as a yellow solid (1.82 g, 84%).
D. 7-[2-Chloro-5-[1-(2-hydroxy-3-piperidin-1-yl-propyl)-5-methanesulfonyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-phenylethynyl]-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester. To a solution of 1-[7-(4-chloro-3-iodo-phenyl)-2-methanesulfonyl-2,3,4,5-tetrahydro-1H-[2]pyridin-5-yl]-3-piperidin-1-yl-propan-2-ol (25 mg, 0.04 mmol), PdCl2(PPh)3 (3 mg, 0.004 mmol), and CuI (1 mg, 0.004 mmol) in degassed THF (1 mL) was added the above alkyne (17 mg, 0.065 mmol) in degassed THF (200 μL), followed by Et3N (200 μL, 0.129 mmol). The reaction mixture was stirred under N2 at 50° C. for 18 h. Additional alkyne (17 mg, 0.065 mmol) in degassed THF (200 μL) was added and the mixture was heated at 50° C. for an additional 1 h. The mixture was diluted with satd. aq. NaHCO3 and extracted with CH2Cl2 (×3). The combined organic extracts were dried (Na2SO4) and concentrated to give a brown oil. Purification by preparative thin layer chromatography (SiO2, 3% MeOH/NH3/CH2Cl2) afforded the desired product as a yellow oil (26 mg, 87%).
E. 1-[3-[4-Chloro-3-(1,2,3,4-tetrahydro-isoquinolin-7-ylethynyl)-phenyl]-5-methanesulfonyl-4,5,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-1-yl}-3-piperidin-1-yl-propan-2-ol. To a solution of 7-{2-chloro-5-[1-(2-hydroxy-3-piperidin-1-yl-propyl)-5-methanesulfonyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-phenylethynyl}-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester (23 mg, 0.033 mmol) in CH2Cl2 (1 mL), was added HCl (1.0 M in Et2O; 100 μL, 0.100 mmol). After 1 h at rt, additional CH2Cl2 (1 mL) and HCl (1.0 M in Et2O; 1 mL, 1.00 mmol) were added and the mixture was stirred for an additional 1 h. HCl (4.0 M in dioxane; 400 μL, 1.6 mmol) was added and the mixture was stirred for an additional 1 h, then was concentrated to afford the desired product. HPLC: Rt=4.31. MS (ESI): mass calculated for C32H38ClN5O3S, 607.2; m/z found, 608.3 [M+H]+. 1H NMR (CD3OD): 7.87 (s, 1H), 7.60 (dd, J=17.9, 8.4, 2H), 7.52-7.46 (m, 2H), 7.32 (d, J=7.9, 1H), 4.53 (s, 2H), 4.50 (br s, 1H), 4.40 (s, 2H), 4.21 (dd, J=12.6, 4.8, 2H), 3.68-3.64 (m, 4H), 3.66 (s, 3H), 3.62-3.50 (m, 4H), 3.18 (dd, J=14.1, 8.2, 3H), 3.11-2.97 (m, 6H), 1.98-1.71 (m, 4H).
A. 1-{3-[3-(4-Chloro-3-iodo-phenyl)-5-methanesulfonyl-4,5,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-1-yl]-propyl}-piperidine-4-carboxylic acid amide. The title compound was prepared using methods analogous to those described in Example 1, Steps A-D, substituting isonipecotamide for morpholine in Step D.
B. 7-(5-[1-[3-(4-Carbamoyl-piperidin-1-yl)-propyl]-5-methanesulfonyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-2-chloro-phenylethynyl)-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester. The title compound was prepared using methods analogous to those described in Example 134, Steps A-D.
C. 1-(3-{3-[4-Chloro-3-(1,2,3,4-tetrahydro-isoquinolin-7-ylethynyl)-phenyl]-5-methanesulfonyl-4,5,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-1-yl-pr piperidine-4-carboxylic acid amide. To a 0° C. solution of 7-(5-{1-[3-(4-carbamoyl-piperidin-1-yl)-propyl]-5-methanesulfonyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl}-2-chloro-phenylethynyl)-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester (80 mg, 0.109 mmol) in CH2Cl2 (3 mL) was added TFA (100 μL, 1.32 mmol). After 30 min at 0° C., additional TFA (1 mL, 13.2 mmol) was added. After 30 min, the mixture was diluted with satd. aq. NaHCO3 and extracted with CH2Cl2. The organic layer was removed and aqueous layer was decanted to leave an oil which was combined with the organic layer and concentrated. The residue was dissolved in MeOH, sonicated, and decanted from precipitates. The MeOH solution was concentrated to afford the desired product (10 mg, 14%). HPLC: Rt=4.14. MS (ESI): mass calcd. for C33H39ClN6O3S, 634.3; m/z found, 635.3 [M+H]+. 1H NMR (CD3OD): 7.86 (d, J=1.8, 1H), 7.59 (dt, J=14.2, 5.2, 2H), 7.53-7.48 (m, 1H), 7.46 (s, 1H), 7.32 (d, J=8.0, 1H), 4.51 (s, 2H), 4.40 (s, 2H), 4.23 (t, J=6.4, 2H), 3.69-3.60 (m, 4H), 3.53 (t, J=6.4, 2H), 3.24-3.13 (m, 4H), 3.04-2.90 (m, 4H), 2.98 (s, 3H), 2.59-2.47 (m, 1H), 2.39-2.28 (m, 2H), 2.07 (d, J=13.9, 2H), 1.96-1.82 (m, 2H).
A. 3-(4-Chloro-3-iodo-phenyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester. This compound was prepared according to the method described in Intermediate 1, Step B, substituting 4-oxo-piperidine-1-carboxylic acid tert-butyl ester for 1-methanesulfonyl-piperidin-4-one. HPLC: Rt=7.69. MS (ESI): mass calcd. for C17H19ClIN3O2, 459.7; m/z found, 460.2 [M+H]+. 1H NMR (CDCl3): 8.08 (br s, 1H), 7.45 (br m, 2H), 4.63 (br s, 2H), 3.73 (br m, 2H), 2.75 (br m, 2H), 1.49 (s, 9H).
B. 3-(4-Chloro-3-iodo-phenyl)-1-(2-cyano-ethyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester. To a solution of the above pyrazole (2.8 g, 6.09 mmol) and acrylonitrile (14 mL) in THF (14 mL) was added 1% aq. NaOH (5.6 mL) dropwise. The reaction mixture was stirred at rt for 8 h, diluted with satd. aq. NH4Cl, and extracted with CH2Cl2 (3×). The combined organic extracts were dried (Na2SO4), filtered, and concentrated to give a brown paste. Recrystallization from EtOH afforded the title compound as a white solid (2.24 g, 72%). HPLC: Rt=7.91. MS (ESI): mass calcd. for C20H22CIIN4O2, 512.8; m/z found, 514.2 [M+H]+. 1H NMR (CDCl3): 8.18 (s, 1H), 7.51-7.46 (br m, 2H), 4.60 (br s, 2H), 4.30 (t, J=6.5, 2H), 3.76-3.72 (br m, 2H), 2.98 (t, J=6.5, 2H), 2.80-2.78 (br m, 2H), 1.49 (s, 9H).
C. 3-(4-Chloro-3-iodo-phenyl)-1-(3-morpholin-4-yl-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester. To a −78 LC solution of the above nitrile (2.39 g, 4.66 mmol) in CH2Cl2 (23 mL) was added DIBAL-H (1.5 M in toluene; 7.75 mL, 5.17 mmol) dropwise. The mixture was stirred at −78 LC for 30 min and then warmed to rt for 30 min. The reaction mixture was cooled to −78 LC and MeOH (15 mL) was added dropwise. The mixture was warmed to rt over 1 h and 1 N H2SO4 (10 mL) was added. After 1 h, the cloudy mixture was diluted with satd. aq. potassium sodium tartrate and extracted with CH2Cl2 (3×). The combined organic extracts were washed with brine, dried (Na2SO4), filtered and concentrated to give a white foam, which was used directly in the next reaction. The reductive amination was conducted in a similar manner to that reported in Intermediate 1, Step D, to provide the title compound as a white solid (1.43 g, 52%). HPLC: Rt=5.42. MS (ESI): mass calcd. for C24H32CIIN4O3, 586.9; m/z found, 588.3 [M+H]+. 1H NMR (CDCl3): 8.18 (d, J=2.1, 1H), 7.54-7.49 (m, 2H), 4.60 (br s, 2H), 4.08 (t, J=6.8, 2H), 3.76-3.71 (br m, 2H), 3.70 (br t, J=4.5, 4H), 2.74 (br t, J=5.4, 2H), 2.43-2.38 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.09-2.01 (m, 2H), 1.49 (s, 9H).
D. 3-[4-Chloro-3-(4-chloro-phenylethynyl)-phenyl]-1-(3-morpholin-4-yl-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester. This compound was prepared from the above iodide according to the method described in Intermediate 1, Step E, substituting 4-chloro-ethynylbenzene for TMSA. HPLC: Rt=6.43. MS (ESI): mass calcd. for C32H36Cl2N4O3, 595.6; m/z found, 596.4 [M+H]+. 1H NMR (CDCl3): 7.85 (s, 1H), 7.65-7.42 (m, 2H), 7.51 (d, J=8.3, 2H), 7.37-7.32 (m, 2H), 4.65 (br s, 2H), 4.09 (t, J=6.8, 2H), 3.78-3.71 (br m, 2H), 3.69 (br t, J=4.5, 4H), 2.75 (br t, J=5.2, 2H), 2.43-2.37 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.10-2.02 (m, 2H), 1.49 (s, 9H).
E. 3-[4-Chloro-3-(4-chloro-phenylethynyl)-phenyl]-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine. To a solution of the above carbamate (1.45 g, 2.43 mmol) in CH2Cl2 (20 mL) was added TFA (5 mL) dropwise. The reaction mixture was stirred at rt for 3 h and then concentrated to give an orange oil. Purification (SiO2; 0-10% 2 M NH3 in MeOH/CH2Cl2) provided the title compound as a clear oil (1.11 g, 92%). HPLC: Rt=4.53. MS (ESI): mass calcd. for C27H28Cl2N4O, 495.5; m/z found, 496.3 [M+H]+. 1H NMR (CDCl3): 7.82 (d, J=2.1, 1H), 7.52-7.49 (m, 3H), 7.43 (d, J=8.4, 1H), 7.34 (dd, J=6.6, 2.0, 2H), 4.10-4.06 (m, 4H), 3.70 (t, J=4.6, 4H), 3.18 (t, J=5.4, 2H), 2.92 (br s, 1H), 2.72 (t, J=5.7, 2H), 2.42 (br m, 4H), 2.34 (t, J=6.9, 2H), 2.09-2.03 (m, 2H).
F. [3-[4-Chloro-3-(4-chloro-phenylethynyl)-phenyl]-1-(3-morpholin-4-yl-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-5-yl]-oxo-acetic acid methyl ester. To a solution of the above amine (100 mg, 0.202 mmol) and pyridine (36 μL, 0.444 mmol) in CH2Cl2 (2.0 mL) was added methyl chlorooxoacetate (20 μL, 0.222 mmol) dropwise. The reaction mixture was stirred at rt for 1.5 h, diluted with satd. aq. NaHCO3, and extracted with CH2Cl2 (3×). The combined organic extracts were dried (Na2SO4), filtered, and concentrated to give a yellow oil. Purification (SiO2; 0-3% 2 M NH3 in MeOH/CH2Cl2) provided the title compound as a clear oil (104 mg, 89%). HPLC: Rt=5.36. MS (ESI): mass calcd. for C30H30Cl2N4O4, 581.5; m/z found, 582.4 [M+H]+. 1H NMR (CDCl3, 2:1 mixture of rotamers): 7.83 (d, J=2.1, 0.7H), 7.78 (d, J=2.0, 0.3H), 7.57-7.40 (m, 4H), 7.35 (dd, J=8.6, 2.0, 2H), 4.83 (s, 1.3H), 4.66 (s, 0.7H), 4.15-4.08 (m, 2H), 3.98 (t, J=5.9, 1H), 3.93 (s, 2H), 3.87 (s, 1H), 3.75-3.69 (m, 5H), 2.91 (t, J=5.6, 1H), 2.86 (t, J=5.8, 1H), 2.40 (br m, 4H), 2.35-2.29 (m, 2H), 2.11-2.06 (m, 2H).
A. 4-Morpholin-4-yl-3,6-dihydro-2H-pyridine-1-carboxylic acid tert-but I ester. In a 3-L round-bottom flask equipped with a Dean-Stark trap, a reflux condenser and an internal thermocouple, N-Boc-piperidone (200 g, 1.0 mol, 1.0 equiv), toluene (2 L), morpholine (92 mL, 1.05 mol, 1.05 equiv), and p-toluenesulfonic acid (1.0 g, 0.005 mmol, 0.5% equiv) were added sequentially. The reaction solution was refluxed under N2 for 16 h (about 18 mL water was collected). The solvent was evaporated and the residue was used directly in the next reaction (colorless oil, ˜270 g, 100%).
B. 4-Chloro-3-iodo-benzoyl chloride. In a 5-L round-bottom flask equipped with a magnetic stirring bar and a gas scrubber, 4-chloro-3-iodobenzoic acid (275 g, 0.975 mol, 1.0 equiv) was suspended in CH2Cl2 (3 L) and DMF (2 mL, 0.026 mol, 2.5% equiv) was added. Under N2 at 0° C., oxalyl chloride (93.5 mL, 1.1 mol, 1.1 equiv) was added dropwise over 1 h. The ice bath was removed and the reaction solution was stirred at rt for 16 h. The solvent was evaporated and the residue was used directly in the next reaction (−290 g, 100%).
C. 3-(4-Chloro-3-iodo-phenyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester. 4-Morpholin-4-yl-3,6-dihydro-2H-pyridine-1-carboxylic acid tert-butyl ester (270 g, 1.0 mol, 1.0 equiv) was dissolved in CH2Cl2 (1.6 L) and then Et3N (209 mL, 1.5 mol, 1.5 equiv) was added. At 0° C. under N2, a solution of 4-chloro-3-iodo-benzoyl chloride (290 g, 1.0 mol, 1.0 equiv) in CH2Cl2 (400 mL) was added over 30 min. The ice bath was then removed and the reaction solution was stirred at rt for 3 h. All the volatile solvents were removed in vacuo and the residue was re-dissolved in EtOH (1.5 L). At 0° C., anhydrous NH2NH2 (47 mL, 1.5 mol, 1.5 equiv) was added over 30 min (exothermic reaction). The reaction solution was stirred at rt for 16 h. The precipitated white solid was collected by filtration and washed with cold EtOH to afford the desired pyrazole product (white solid, ˜333 g, 0.73 mol, >95% purity, 73%). The mother liquor was concentrated and was partitioned between CH2Cl2 and H2O. Emulsion was observed due to the low solubility of the desired product in CH2Cl2. The insoluble solid was collected by filtration to provide an additional portion of the desired product. The organic layer was warmed slightly to assist phase separation. The organic layer was washed with water (3×), dried, and concentrated. The crude product (filtered material plus residue) was recrystallized from hot CH3CN to give the title compound (74 g, 0.16 mmol, 16%). The total yield was 89% for the three steps.
The compounds in Examples 137-142 were prepared using methods analogous to those described in Example 136, substituting the appropriate acid chloride for methyl chlorooxoacetate in Step F.
HPLC: Rt=5.48. MS (ESI): mass calcd. for C33H31Cl2N5O2, 600.6; m/z found, 601.5 [M+H]+. 1H NMR (CDCl3): 8.60 (t, J=4.8, 1H), 7.90 (d, J=2.0, 1H), 7.87-7.77 (m, 1H), 7.76-7.70 (m, 1H), 7.63 (dd, J=8.4, 2.1, 1H), 7.56-7.46 (m, 3H), 7.42-7.33 (m, 4H), 4.99 (s, 1H), 4.14-4.08 (m, 3H), 3.86 (t, J=5.6, 1H), 3.72-3.68 (m, 4H), 2.98-2.90 (m, 2H), 2.44-2.38 (br m, 4H), 2.37-2.31 (m, 2H), 2.13-2.05 (m, 2H).
HPLC: Rt=5.27. MS (ESI): mass calcd. for C32H30Cl2N4O3, 589.5; m/z found, 590.4 [M+H]+. 1H NMR (CDCl3): 7.86 (s, 1H), 7.54-7.50 (m, 3H), 7.46 (d, J=8.4, 2H), 7.37-7.33 (m, 2H), 7.08 (d, J=3.2, 1H), 6.52-6.50 (m, 1H), 4.95 (br s, 2H), 4.11 (t, J=6.9, 2H), 4.05 (t, J=5.6, 2H), 3.70 (t, J=4.6, 4H), 2.96-2.90 (br m, 2H), 2.45-2.39 (br m, 4H), 2.34 (t, J=6.8, 2H), 2.12-2.06 (m, 2H).
MS (ESI): mass calcd. for C29H27Cl2F3N4O2, 591.5; m/z found, 592.4 [M+H]+.
MS (ESI): mass calcd. for C29H29Cl2FN4O2, 555.5; m/z found, 556.4 [M+H]+.
MS (ESI): mass calcd. for C32H34Cl2N4O3, 593.6; m/z found, 594.4 [M+H]+.
MS (ESI): mass calcd. for C31H32Cl2N4O4, 595.5; m/z found, 596.4 [M+H]+.
To a slurry of acetic acid 2-[3-[4-chloro-3-(4-chloro-phenylethynyl)-phenyl]-1-(3-morpholin-4-yl-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-5-yl]-2-oxo-ethyl ester (105 mg, 0.176 mmol) in 2:1 MeOH/H2O (1.5 mL) was added NaOH (14 mg, 0.352 mmol). The reaction mixture was stirred at rt for 2 h, diluted with satd. aq. NaHCO3, and extracted with CH2Cl2 (3×). The combined organic extracts were dried (Na2SO4), filtered, and concentrated to give a yellow oil. Purification (SiO2; 0-5% 2 M NH3 in MeOH/CH2Cl2) provided the title compound as a white solid (62 mg, 64%). MS (ESI): mass calcd. for C29H30Cl2N4O3, 553.5; m/z found, 554.4 [M+H]+.
This compound was prepared according to the methods described in Example 136, substituting trimethylsilyl isocyanate for methyl chlorooxoacetate in Step F. HPLC: Rt=4.81. MS (ESI): mass calcd. for C28H29Cl2N5O2, 538.5; m/z found, 539.3 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=2.0, 1H), 7.52-7.50 (m, 2H), 7.43 (d, J=8.4, 2H), 7.34 (d, J=8.5, 2H), 4.90 (s, 2H), 4.58 (s, 2H), 4.12-4.06 (m, 2H), 3.74 (t, J=5.6, 2H), 3.68 (t, J=4.5, 4H), 2.77 (t, J=5.4, 2H), 2.39 (br m, 4H), 2.32 (t, J=6.8, 2H), 2.08-1.99 (m, 2H).
The compounds in Examples 145-149 were prepared using methods analogous to those described in Example 2. The alkynes used as starting materials for Examples 145-149 are described in the preceding examples.
HPLC: Rt=5.04. MS (ESI): mass calcd. for C28H33Cl2N5O2, 542.5; m/z found, 543.5 [M+H]+. 1H NMR (CDCl3): 7.43 (d, J=1.8, 1H), 7.36 (dd, J=8.3, 2.0, 2H), 7.26-7.23 (m, 2H), 7.16-7.13 (m, 2H), 4.61 (br s, 2H), 4.47 (s, 2H), 4.09 (t, J=6.9, 2H), 3.76 (t, J=5.7, 2H), 3.69 (t, J=4.6, 4H), 3.06-3.01 (m, 2H), 2.95-2.90 (m, 2H), 2.78 (t, J=5.7, 2H), 2.42-2.38 (br m, 4H), 2.33 (t, J=6.8, 2H), 2.13-2.05 (m, 2H).
HPLC: Rt=5.33. MS (ESI): mass calcd. for C34H41ClF3N5O4S, 708.3; m/z found, 709.5 [M+H]+. 1H NMR (CDCl3): 7.68 (d, J=8.2, 1H), 7.54 (s, 1H), 7.46 (d, J=8.2, 1H), 7.28-7.25 (m, 2H), 7.17-7.13 (m, 2H), 4.49 (dd, J=18.5, 14.6, 2H), 4.19 (dd, J=13.8, 2.8, 1H), 4.16-4.09 (m, 1H), 4.04-3.96 (m, 2H), 3.73-3.62 (m, 2H), 3.34 (t, J=7.0, 2H), 3.13-2.99 (m, 4H), 2.95-2.86 (m, 5H), 2.88 (s, 3H), 2.50-2.42 (m, 3H), 2.39 (t, J=8.2, 2H), 2.05-1.97 (m, 3H), 1.76-1.62 (m, 4H).
HPLC: Rt=5.56. MS (ESI): mass calcd. for C29H34ClF3N4O4S, 627.1; m/z found, 628.4 [M+H]+. 1H NMR (CDCl3): 7.68 (d, J=8.2, 1H), 7.53 (s, 1H), 7.46 (d, J=8.2, 1H), 7.28-7.25 (m, 2H), 7.17-7.14 (m, 2H), 4.49 (dd, J=19.4, 14.5, 2H), 4.21 (dd, J=13.7, 2.8, 1H), 4.18-4.13 (m, 1H), 4.02 (dd, J=13.6, 6.7, 1H), 3.75-3.62 (m, 6H), 3.12-3.02 (m, 3H), 2.96-2.87 (m, 4H), 2.88 (s, 3H), 2.66-2.60 (m, 2H), 2.50-2.40 (m, 4H).
HPLC: Rt=5.21. MS (ESI): mass calcd. for C32H39Cl2N5O4S, 660.7; m/z found, 661.5 [M+H]+. 1H NMR (CDCl3): 7.40-7.30 (m, 4H), 7.23 (d, J=7.5, 2H), 7.13 (d, J=7.9, 2H), 4.43 (dd, J=17.8, 15.0, 2H), 4.17-4.10 (m, 2H), 4.01-3.94 (m, 1H), 3.70-3.60 (m, 2H), 3.35-3.33 (m, 1H), 3.30 (t, J=6.8, 2H), 3.07-2.99 (m, 3H), 2.93-2.78 (m, 5H), 2.87 (s, 3H), 2.45-2.38 (m, 2H), 2.30 (br t, J=10.8, 2H), 2.10 (br t, J=11.7, 2H), 2.02 (t, J=6.9, 2H), 1.95-1.82 (m, 2H).
HPLC: Rt=5.47. MS (ESI): mass calcd. for C28H30Cl2N4O4S, 589.6; m/z found, 590.2 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.5, 1H), 7.54-7.50 (m, 2H), 7.48-7.46 (m, 2H), 7.36-7.33 (m, 2H), 4.53 (dd, J=18.3, 14.5, 2H), 4.18 (dd, J=13.3, 2.9, 1H), 4.17-4.10 (m, 1H), 3.99 (dd, J=13.7, 6.6, 1H), 3.71-3.60 (m, 6H), 3.07-2.99 (m, 1H), 2.93-2.85 (m, 2H), 2.87 (s, 3H), 2.65-2.58 (m, 2H), 2.48-2.38 (m, 4H).
Unless otherwise specified, the compounds in Examples 150-621 were prepared as free base, hydrochloride salt, trifluoroacetic acid salt, citric acid, or formic acid salt forms.
A. 3-(4-Chloro-3-iodo-phenyl)-1-(3-hydroxy-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester. A mixture of 3-(4-chloro-3-iodo-phenyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester (21.0 mmol) and Cs2CO3 (43.8 mmol) in DMF (73 mL) at 0° C., was treated with 3-bromo-propan-1-ol (32.9 mmol). The mixture was stirred at rt for 15 h. The mixture was poured into water and extracted with EtOAc (3×). The combined organic layers were washed satd. aq. NaCl (4×). The organic phase was dried and concentrated, and the resulting oil was purified (SiO2; 10-50% EtOAc/hexanes) to afford the title compound (74%). MS (ESI): mass calcd. for C20H25CIIN3O3, 517.06; m/z found, 518.1 [M+H]+.
B. 3-(4-Chloro-3-iodo-phenyl)-1-(3-oxo-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester. A solution of the alcohol above (9.8 mmol) and Dess-Martin periodinane (14.7 mmol) in CH2Cl2 (43.4 mL) was stirred for 2 h at rt. The mixture was poured into satd. aq. NaHCO3 and extracted with CH2Cl2 (3×). The combined organic layers were dried and concentrated. The resulting oil was passed through a plug of SiO2 eluting with CH2Cl2 and EtOAc. The filtrate was concentrated and used immediately. MS (ESI): mass calcd. for C20H23CIIN3O3, 515.05; m/z found, 516.1 [M+H]+.
C. 3-(4-Chloro-3-iodo-phenyl)-1-(3-morpholin-4-yl-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester. This compound was prepared from the aldehyde above according to the method described for Intermediate 1, Part D. MS (ESI): mass calcd. for C24H32CIIN4O3, 586.12; m/z found, 587.2 [M+H]+
D. 3-[4-Chloro-3-(4-{[(4-chloro-benzyl)-(2,2,2-trifluoro-acetyl)-amino]-methyl]-phenylethynyl)-phenyl]-1-(3-morpholin-4-yl-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester. This compound was prepared according to the method described for Intermediate 1, Part E, substituting N-(4-chloro-benzyl)-N-(4-ethynyl-benzyl)-2,2,2-trifluoro-acetamide for (trimethylsilyl)acetylene. MS (ESI): mass calcd. for C42H44Cl2F3N5O4, 809.27; m/z found, 810.2 [M+H]+.
E. N-(4-Chloro-benzyl)-N-(4-{2-chloro-5-[1-(3-morpholin-4-yl-prolyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl phenylethynyl]-benzyl)-2,2,2-trifluoro-acetamide. This compound was prepared according to the method described in Example 136, Part E. MS (ESI): mass calcd. for C37H36Cl2F3N5O2, 709.22; m/z found, 710.2 [M+H]+.
F. N-(4-[5-[5-Aminooxalyl-1-(3-morpholin-4-propyl)-3a,4,5,6,7,7a-hexahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-2-chloro-phenylethynyl]-benzyl)-N-(4-chloro-benzyl)-2,2,2-trifluoro-acetamide. To N-(4-chloro-benzyl)-N-(4-{2-chloro-5-[1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenylethynyl}-benzyl)-2,2,2-trifluoro-acetamide (0.132 mmol) was added oxalamic acid (0.527 mmol), HATU (0.264 mmol), HOAT (0.5 M in DMF; 530 μL), DMF (0.66 mL), and iPr2NEt (0.396 mmol). The mixture was stirred at 80° C. for 26.5 h. The mixture was poured into satd. aq. NaHCO3 (10 mL). The aqueous phase was extracted CH2Cl2 (3×10 mL) and EtOAc (2×10 mL). The combined organic layers were dried and concentrated. The resulting oil was purified (SiO2; 0-5% 0.2 M NH3 in MeOH/CH2Cl2) to afford the title compound as an oil (60%, 0.080 mmol). MS (ESI): mass calcd. for C39H39Cl2F3N6O4, 782.24; m/z found, 783.2 [M+H]+.
G. 2-[3-(4-Chloro-3-{[4-(ff(4-chlorophenyl)methyl]amino]methyl)phenyl]ethynyl}phenyl)-1-(3-morpholin-4-ylpropyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl]-2-oxoacetamide. A mixture of N-(4-{5-[5-aminooxalyl-1-(3-morpholin-4-yl-propyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-2-chloro-phenylethynyl}-benzyl)-N-(4-chloro-benzyl)-2,2,2-trifluoro-acetamide (0.026 mmol) and K2CO3 (0.153 mmol) in MeOH/H2O (2:1, 1.5 mL) was stirred for 30 min. The mixture was poured into H2O, extracted with CH2Cl2 (2×) and EtOAc (2×). The combined organic layers were dried and concentrated. The resulting oil was purified using preparatory HPLC (Method E). MS (ESI): mass calcd. for C37H38Cl2N6O3, 684.24; m/z found, 685.2 [M+H]+. 1H NMR: 7.92-7.81 (m, 1H), 7.68-7.63 (m, 3H), 7.60-7.56 (m, 3H), 7.55-7.52 (m, 2H), 7.50-7.44 (m, 2H), 4.80 (s, 1H), 4.32-4.23 (m, 6H), 4.05-3.91 (m, 4H), 3.78 (t, J=12, 2H), 3.51 (d, J=12, 2H), 3.31-3.23 (m, 6H), 3.15 (dt, J=12, 3, 2H), 3.00-2.87 (m, 2H), 2.42-2.33 (m, 2H).
The compounds in Examples 151-155 were prepared using methods analogous to those described in Example 150, substituting oxalamic acid with the appropriate carboxylic acids (Step F), or substituting oxalamic acid, HATU, HOAt, iPr2NEt, and DMF with the appropriate sulfonyl chlorides, acid chlorides, or isocyanates in CH2Cl2, with exceptions or alterations where noted.
The compound was purified using preparatory HPLC (Method E; HCl salt). MS (ESI): mass calcd. for C36H38Cl2N6O2, 656.24; m/z found, 657.2 [M+H]+. 1H NMR: 7.93 (d, J=1.9, 1H), 7.68-7.64 (m, 3H), 7.63-7.59 (m, 3H), 7.59-7.54 (m, 3H), 7.49-7.45 (m, 2H), 4.76 (s, 1H), 4.35-4.25 (m, 7H), 4.02 (dd, J=13, 2.9, 2H), 3.90 (t, J 5.5, 2H), 3.84 (dd, J=25, 13, 2H), 3.52 (d, J=12, 2H), 3.30-3.25 (m, 4H), 3.17 (dt, J=12, 3.4, 2H), 2.96 (t, J=5.2, 2H), 2.46-2.36 (m, 2H).
The compound was purified using preparatory HPLC (Method F; HCl salt). MS (ESI): mass calcd. for C39H42Cl2N6O3, 712.27; m/z found, 713.3 [M+H]+. 1H NMR: 7.89 (m, 1H), 7.70-7.65 (m, 3H), 7.63-7.58 (m, 3H), 7.57-7.54 (m, 2H), 7.52-7.47 (m, 2H), 4.87-4.85 (m, 1H), 4.32 (d, J=15, 4H), 4.28 (t, J=6.6, 2H), 4.07-4.01 (m, 3H), 3.86-3.74 (m, 4H), 3.52 (d, J=13, 2H), 3.31-3.24 (m, 2H), 3.17 (dt, J=12, 3.6, 2H), 3.07 (d, J=11, 5H), 3.01-2.92 (m, 4H), 2.44-2.35 (m, 2H).
The compound was purified using preparatory HPLC (Method F; HCl salt). MS (ESI): mass calcd. for C37H42Cl2N6O3S, 720.24; m/z found, 721.3 [M+H]+. 1H NMR (DMSO): 7.90 (d, J=1.7, 1H), 7.71-7.62 (m, 7H), 7.60 (dd, J=8.4, 1.7, 1H), 7.51 (d, J=8.3, 2H), 4.49 (s, 2H), 4.22-4.13 (m, 6H), 3.93 (d, J=11, 2H), 3.88-3.84 (m, 3H), 3.58 (t, J=5.3, 2H), 3.41 (d, J=12, 2H), 3.19-3.11 (m, 2H), 3.11-2.99 (m, 2H), 2.90-2.86 (m, 2H), 2.80-2.73 (m, 6H), 2.34-2.27 (m, 2H).
The compound was purified using preparatory HPLC (Method E). Collected fractions were concentrated and the residue was treated with HCl (1.25 N in MeOH; 3 mL) in CH2Cl2 (3 mL). After 18 h, the mixture was concentrated to give the compound as the HCl salt. MS (ESI): mass calcd. for C37H40Cl2N6O2, 670.26; m/z found, 671.3 [M+H]+. 1H NMR: 7.99-7.89 (m, 1H), 7.72-7.64 (m, 3H), 7.63-7.59 (m, 3H), 7.57-7.52 (m, 2H), 7.51-7.48 (m, 2H), 4.86 (s, 2H), 4.70 (d, J=18, 1H), 4.36-4.25 (m, 6H), 4.17 (s, 1.3H), 4.11 (s, 0.7H), 4.04 (d, J=13, 3H), 3.89-3.75 (m, 4H), 3.54 (d, J=12, 2H), 3.31-3.24 (m, 3H), 3.18 (t, J=11, 2H), 3.05 (s, 1.3H), 2.92 (s, 0.7H), 2.41 (d, J=1.0, 2H).
The compound was purified using preparatory HPLC (Method F; HCl salt). MS (ESI): mass calcd. for C37H40Cl2N6O2, 670.26; m/z found, 671.3 [M+H]+. 1H NMR: 7.84 (d, J=2.1, 1H), 7.61-7.54 (m, 3H), 7.53-7.48 (m, 3H), 7.47-7.45 (m, 2H), 7.40-7.36 (m, 2H), 4.94 (s, 1H), 4.54 (s, 2H), 4.24-4.18 (m, 6H), 3.93 (dd, J=13, 3.2, 2H), 3.75-3.67 (m, 4H), 3.42 (d, J=13, 2H), 3.20-3.16 (m, 2H), 3.07 (dt, J=12, 3.6, 2H), 2.77 (t, J=5.6, 2H), 2.66 (s, 3H), 2.35-2.25 (m, 2H).
This compound was prepared according to the methods described in Example 150, following sequentially Steps A, B, C, E, F, and then D with the appropriate substituent changes. The compound was purified using preparatory HPLC (Method B; TFA salt). MS (ESI): mass calcd. for C41H42Cl2N8O, 732.29; m/z found, 733.3 [M+H]+. 1H NMR: 8.17 (t, J=7.7, 1H), 8.08 (d, J=5.8, 1H), 7.96-7.93 (m, 1H), 7.69 (d, J=7.9, 3H), 7.64-7.57 (m, 3H), 7.57-7.53 (m, 2H), 7.53-7.45 (m, 3H), 7.17 (t, J=6.5, 1H), 4.73 (s, 2H), 4.53-4.36 (m, 2H), 4.31 (d, J=12.2, 6H), 4.01-3.55 (m, 8H), 3.43-3.34 (m, 3H), 2.98-2.91 (m, 2H), 2.52-2.43 (m, 2H), 1.43-1.25 (m, 2H).
The compounds in Examples 157-165 were prepared using methods analogous to those described for Example 156.
The compound was purified using preparatory HPLC (Method B; HCl salt). MS (ESI): mass calcd. for C39H42Cl2N6O3, 712.27; m/z found, 714.2 [M+H]+. 1H NMR: 7.93-7.83 (m, 1H), 7.71-7.67 (m, 3H), 7.60 (t, J=9.3, 3H), 7.55-7.49 (m, 4H), 4.93-4.90 (m, 1H), 4.84-4.81 (m, 1H), 4.31 (d, J=13.2, 4H), 4.28-4.24 (m, 3H), 4.06-3.93 (m, 3H), 3.89 (q, J=13.0, 2H), 3.77-3.69 (m, 1H), 3.69-3.60 (m, 1H), 3.62-3.42 (m, 2H), 3.38-3.35 (m, 2H), 3.22-3.08 (m, 1H), 3.09-2.87 (m, 2H), 2.50-2.36 (m, 1H), 2.31-2.19 (m, 1H), 1.42 (d, J=6.5, 3H), 1.30 (d, J=6.1, 3H).
The compound was purified using preparatory HPLC (Method B; HCl salt). MS (ESI): mass calcd. for C38H40Cl2N6O3, 698.25; m/z found, 699.2 [M+H]+. 1H NMR: 7.90-7.89 (m, 0.7H), 7.84-7.82 (m, 0.3H), 7.70-7.66 (m, 3H), 7.61-7.55 (m, 3H), 7.53-7.45 (m, 4H), 4.81 (s, 1H), 4.31 (s, 2H), 4.28 (s, 2H), 4.27-4.22 (m, 2H), 4.05 (d, J=13, 1H), 4.02-3.94 (m, 3H), 3.87-3.68 (m, 2H), 3.58-3.39 (m, 4H), 3.35-3.32 (m, 1H), 3.26-3.13 (m, 3H), 3.02-2.85 (m, 2H), 2.47-2.35 (m, 1H), 2.33-2.20 (m, 1H), 1.45-1.22 (m, 4H).
A. 4-(3-Chloro-propyl}-(3S)-methyl-morpholine. A solution of (3S)-methyl-morpholine (1 g, 9.9 mmol, 1.0 equiv) and 1-bromo-3-chloro-propane (3.1 g, 19.8 mmol, 2.0 equiv) in THF (5 mL) was treated with NaH (60%, 2 equiv) in two portions. The resulting slurry was heated at 65° C. for 18 h. Slowly, the reaction was quenched with ice water (20 mL). During the addition, excess bubbling occurred. The resulting mixture was stirred for 16 h and then was extracted with EtOAc (3×15 mL). The combined organic layers were extracted with 1 N HCl (40 mL). The aqueous layer was cooled in an ice bath and basified slowly to pH ˜9-10 with NaOH pellets. The aqueous layer was extracted with EtOAc (3×15 mL). The combined organic layers were dried and concentrated to give the title compound as a clear oil (1.3 g, 74%).
B. 3-(4-Chloro-3-iodo-phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine. A suspension of 3-(4-chloro-3-iodo-phenyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester (260 g, 0.57 mol, 1.0 equiv) in CH2Cl2 (750 mL) was treated with TFA (250 mL) over 20 min. The resulting solution was stirred at rt for 16 h. The mixture was diluted with water (2 L) and then basified with satd. aq. NaOH to a pH>12. The mixture was stirred for 3 h. The white precipitate was collected by filtration, washed with water and dried in a vacuum oven to provide the title compound (205 g, 0.57 mol, 100%), which was used in the next reaction without further purification.
C. 2-[3-(4-Chloro-3-iodo-phenyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-5-yl]-2-oxo-acetamide. A suspension of CDI (110.5 g, 0.68 mol, 1.2 equiv) in DMF (1.5 L) was treated with oxalamic acid (60.7 g, 0.68 mol, 1.2 equiv). After 3 h at rt, 3-(4-chloro-3-iodo-phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine (205 g, 0.57 mol, 1.0 equiv) was added as solid over 10 min. After 20 min, water (2.5 L) was added slowly over 2 h and the mixture was stirred at rt for 16 h. The resulting white precipitate was collected by filtration, washed with water and dried in a vacuum oven to provide the title compound (243 g, 0.56 mol, 100%), which was used in the next reaction without further purification.
D. 2-[3-(4-Chloro-3-iodo-phenyl)-1-[3-((3S)-methyl-morpholin-4-yl)-propyl]-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-5-yl]-2-oxo-acetamide. A solution of 2-[3-(4-chloro-3-iodo-phenyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-5-yl]-2-oxo-acetamide (190 g, 0.44 mol, 1.0 equiv) in DMF (1.8 L) was treated with Cs2CO3 (180 g, 0.55 mol, 1.25 equiv) followed by 4-(3-chloro-propyl)-(3S)-methyl-morpholine (86 g, 0.48 mol, 1.1 equiv). The reaction mixture was stirred via mechanical stirrer at 50° C. under N2 for 12 h and then cooled to rt. Water (3.5 L) was added slowly over 30 min. The mixture was stirred at rt for 16 h and the resulting white solid was collected by filtration, washed with water and dried in a vacuum oven. The crude compound was triturated from hot EtOH (−1.5 L) to provide the title compound (176 g, 0.31 mol, 70%).
E. 2-[3-(4-Chloro-3-{[4-(ff(4-chlorophenyl)methyl]amino]methyl)phenyl]ethynyl}phenyl)-1-{3-[(3S)-3-methylmorpholin-4-yl]propyl-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl-2-oxoacetamide. To a 5 L flask equipped with a mechanical stirrer and an internal thermocouple, was added 2-{3-(4-chloro-3-iodo-phenyl)-1-[3-((3S)-methyl-morpholin-4-yl)-propyl]-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-5-yl}-2-oxo-acetamide (120 g, 0.21 mol, 1.0 equiv), (4-chloro-benzyl)-(4-ethynyl-benzyl)-amine (56.3 g, 0.22 mol, 1.0 equiv), DMF (1.5 L) and Et3N (120 mL, 0.84 mol, 4.0 equiv) sequentially. A stream of N2 was bubbled into the solution for 15 min. A mixture of Pd(PPh3)2Cl2 (0.37 g, 0.5 mmol, 0.0025 equiv) and CuI (0.2 g, 1.0 mmol, 0.005 equiv) was added under N2. The solution was degassed with N2 for another 10 min. The reaction solution was stirred at 50° C. for 16 h. The reaction solution was cooled to rt and water (2 L) was added with stirring. The liquid layer was decanted away from an oily precipitate, which was then partitioned between EtOAc (2 L) and 2:1 water/satd. aq. NaHCO3 (1.5 L). The organic layer was dried and concentrated to provide the crude material as a foamy yellow solid (145 g, ˜85% purity by HPLC). The crude material was purified (SiO2; 2 N NH3 in MeOH/CH2Cl2) to provide the title compound (87 g, 59%, >98% purity).
The compound was purified using preparatory HPLC (Method B; HCl salt). MS (ESI): mass calcd. for C39H40Cl2N6O3, 710.25; m/z found, 711.2 [M+H]+. 1H NMR: 7.80-7.69 (m, 1H), 7.59-7.54 (m, 1H), 7.54-7.38 (m, 6H), 7.35 (s, 1H), 7.25 (d, J=7.9, 1H), 4.70 (s, 2H), 4.43 (s, 2H), 4.36 (d, J=7.3, 2H), 4.20-4.09 (m, 2H), 4.01-3.77 (m, 4H), 3.72 (s, 1H), 3.64 (t, J=12.0, 2H), 3.40 (d, J=12.8, 3H), 3.18-3.12 (m, 5H), 3.11-2.96 (m, 3H), 2.87 (s, 1H), 2.83-2.78 (m, 1H), 2.34-2.14 (m, 2H).
The compound was purified using preparatory HPLC (Method B; HCl salt). MS (ESI): mass calcd. for C36H38Cl2N6OS, 672.22; m/z found, 673.3 [M+H]+. 1H NMR: 7.92-7.90 (m, 1H), 7.70-7.65 (m, 3H), 7.60-7.55 (m, 3H), 7.53-7.47 (m, 4H), 4.93-4.90 (m, 1H), 4.66 (s, 2H), 4.29 (d, J=16.2, 4H), 4.24 (t, J=6.4, 2H), 3.85-3.74 (m, 4H), 3.35-3.33 (m, 1H), 3.28-3.17 (m, 5H), 3.06 (t, J=13.3, 2H), 2.87-2.81 (m, 4H), 2.38-2.31 (m, 2H).
The compound was purified using preparatory HPLC (Method B; HCl salt). MS (ESI): mass calcd. for C35H37ClN6O2S, 640.24; m/z found, 641.2 [M+H]+. 1H NMR: 8.89 (s, 1H), 8.81 (d, J=5.4, 1H), 8.67 (d, J=7.9, 1H), 8.11 (dd, J=7.8, 6.0, 1H), 7.91 (s, 1H), 7.65 (d, J=8.3, 1H), 7.62-7.56 (m, 3H), 7.51-7.47 (m, 2H), 4.85 (s, 2H), 4.77 (s, 2H), 4.67 (s, 2H), 4.27-4.23 (m, 2H), 3.87-3.74 (m, 4H), 3.28-3.17 (m, 5H), 3.14-3.03 (m, 2H), 2.89-2.79 (m, 4H), 2.36 (d, J=1.2, 2H), 1.29-1.28 (m, 1H).
The compound was purified using preparatory HPLC (Method B; HCl salt). MS (ESI): mass calcd. for C35H36ClN7O2S, 653.23; m/z found, 654.2 [M+H]+. 1H NMR: 8.92 (s, 1H), 8.80 (d, J=5.2, 1H), 8.68 (d, J=8.0, 1H), 8.11 (dd, J=7.9, 5.8, 1H), 7.99-7.90 (m, 3H), 7.73-7.65 (m, 3H), 7.59 (d, J=8.3, 1H), 4.83-4.74 (m, 3H), 4.70 (s, 2H), 4.30-4.20 (m, 2H), 3.82 (d, J=10.3, 3H), 3.35 (s, 4H), 3.28-3.17 (m, 4H), 3.17-3.06 (m, 2H), 2.95-2.73 (m, 3H), 2.43-2.30 (m, 2H).
The compound was purified using preparatory HPLC (Method B; HCl salt). MS (ESI): mass calcd. for C35H43ClN6O2S, 646.29; m/z found, 647.3 [M+H]+. 1H NMR: 7.91 (s, 1H), 7.65 (dd, J=8.4, 2.1, 1H), 7.57 (dd, J=9.6, 8.4, 3H), 7.40 (d, J=8.1, 2H), 4.93 (s, 1H), 4.66 (s, 2H), 4.56 (s, 2H), 4.24 (t, J=6.5, 2H), 3.85-3.75 (m, 4H), 3.47-3.36 (m, 5H), 3.28-3.17 (m, 5H), 3.14-3.03 (m, 2H), 3.00 (t, J=11.8, 2H), 2.88-2.79 (m, 4H), 2.42-2.28 (m, 2H), 2.03-1.96 (m, 3H), 1.57-1.48 (m, 2H).
The compound was purified using preparatory HPLC (Method B; HCl salt). MS (ESI): mass calcd. for C33H37ClN6O2S, 616.24; m/z found, 617.2 [M+H]+. 1H NMR (DMSO): 7.93 (d, J=1.6, 1H), 7.70 (dd, J=8.5, 1.7, 1H), 7.67 (d, J=8.5, 3H), 7.60 (d, J=8.1, 2H), 4.57 (s, 2H), 4.14 (t, J=6.8, 2H), 3.71 (d, J=8.4, 2H), 3.64 (t, J=5.4, 2H), 3.48 (t, J=6.7, 2H), 3.44-3.31 (m, 4H), 3.20-3.09 (m, 6H), 2.81 (d, J=13.1, 2H), 2.73 (t, J=4.5, 2H), 2.31-2.18 (m, 2H), 1.88 (td, J=13.0, 6.4, 2H), 1.85-1.79 (m, 2H).
The compound was purified using preparatory HPLC (Method B; HCl salt). MS (ESI): mass calcd. for C38H46ClN7O2S, 699.31; m/z found, 700.3 [M+H]+. 1H NMR: 7.95 (d, J=2.0, 1H), 7.72 (d, J=8.2, 2H), 7.69 (dd, J=8.4, 2.1, 1H), 7.65 (d, J=8.2, 2H), 7.60 (d, J=8.4, 1H), 4.69 (s, 2H), 4.39 (s, 2H), 4.26 (t, J=6.5, 2H), 4.21-4.08 (m, 1H), 3.86-3.80 (m, 4H), 3.60 (d, J=12.6, 2H), 3.46 (t, J=7.0, 2H), 3.30-3.05 (m, 9H), 2.91-2.80 (m, 4H), 2.39 (dd, J=15.2, 6.9, 4H), 2.21-2.10 (m, 2H), 2.10-2.01 (m, 2H), 2.00-1.92 (m, 2H), 1.30 (s, 1H).
The title compound was prepared according to the methods describe for Example 150, but the reaction described in Step G proceeded for 1 h. The compound was purified using preparatory HPLC (Method B; TFA salt). MS (ESI): mass calcd. for C35H37Cl2N5O, 613.24; m/z found, 614.2 [M+H]+. 1H NMR: 7.88 (s, 1H), 7.66 (d, J=8.1, 2H), 7.60-7.58 (m, 2H), 7.56 (d, J=8.2, 2H), 7.52-7.47 (m, 4H), 4.94-4.91 (m, 2H), 4.48 (s, 2H), 4.33-4.23 (m, 6H), 4.12-3.91 (m, 2H), 3.87-3.67 (m, 2H), 3.61 (t, J=6.1, 2H), 3.57-3.42 (m, 2H), 3.34 (s, 1H), 3.25 (dd, J=9.4, 6.7, 2H), 3.16 (t, J=6.0, 3H), 2.41-2.34 (m, 2H).
A. 3-(4-Chloro-3-iodo-phenyl)-1-(3-piperidin-1-yl-prolyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid amide. This compound was prepared according to the methods described in Example 150, following sequentially Steps A, B, C, E, and then F, with the appropriate substituent changes. MS (ESI): mass calcd. for C21H27CIIN5O, 527.09; m/z found, 528.1 [M+H]+.
B. 3-(4-Chloro-3-trimethylsilanylethynyl-phenyl)-1-(3-piperidin-1-yl-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid amide. This compound was prepared according to the methods described for Intermediate 1, Step E. MS (ESI): mass calcd. for C26H36ClN5OSi, 497.24; m/z found, 498.2 [M+H]+.
C. 3-[4-Chloro-3-({4-chloro-3-[(ethylamino)methyl]phenyl}ethynyl)phenyl]-1-(3-piperidin-1-ylpropyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]oyridine-5-carboxamide. This compound was prepared according to the methods described for Intermediate 1, Step E, substituting (2-chloro-5-iodo-benzyl)-ethyl-amine for (trimethylsilyl)acetylene, DMF instead of THF, DBU instead of TEA, and adding 2.8 eq. H2O. The compound was purified using preparatory HPLC (Method C; HCl salt). MS (ESI): mass calcd. for C32H38Cl2N6O, 592.25; m/z found, 593.1 [M+H]+. 1H NMR: 8.02-7.78 (m, 2H), 7.70-7.55 (m, 4H), 4.76-4.67 (m, 2H), 4.46-4.37 (m, 2H), 4.30-4.22 (m, 2H), 3.94-3.79 (m, 2H), 3.54 (d, J=9.2, 2H), 3.29-3.13 (m, 4H), 3.03-2.85 (m, 4H), 2.45-2.30 (m, 2H), 1.92 (d, J=12.6, 2H), 1.80 (d, J=11.5, 4H), 1.59-1.46 (m, 1H), 1.43-1.38 (m, 4H), 1.31-1.26 (m, 1H).
The compounds in Examples 168-169 were prepared using methods analogous to those described in Example 167, with the appropriate substituent changes.
The compound was purified using preparatory HPLC (Method B; TFA salt). MS (ESI): mass calcd. for C37H43Cl2N7O5, 735.27; m/z found, 736.3 [M+H]+. 1H NMR: 7.88-7.79 (m, 1H), 7.74 (d, J=1.9, 1H), 7.66 (dd, J=8.4, 2.1, 1H), 7.62 (dd, J=8.3, 2.0, 1H), 7.58-7.52 (m, 2H), 4.83-4.76 (m, 1H), 4.25-4.19 (m, 2H), 4.17-4.00 (m, 3H), 3.99-3.91 (m, 4H), 3.87-3.70 (m, 5H), 3.69-3.59 (m, 2H), 3.59-3.49 (m, 3H), 3.49-3.42 (m, 3H), 3.28-3.12 (m, 5H), 2.96-2.92 (m, 1.3H), 2,89-2.85 (m, 0.7H), 2.47-2.32 (m, 1H), 2.32-2.18 (m, 1H), 1.39 (s, 1H), 1.27 (d, J=5.6, 2H).
The compound was purified using preparatory HPLC (Method D; formic acid salt). MS (ESI): mass calcd. for C37H44ClN7O5, 701.31; m/z found, 702.2 [M+H]+.
1H NMR: 7.92-7.80 (m, 3H), 7.66 (d, J=8.4, 2H), 7.63 (dd, J=8.4, 2.1, 1H), 7.56-7.54 (m, 1H), 4.88-4.85 (m, 2H), 4.79 (s, 2H), 4.14 (t, J=5.9, 2H), 3.95 (td, J=19.4, 5.8, 2H), 3.85-3.79 (m, 1H), 3.73 (dd, J=11.8, 2.7, 1H), 3.69-3.64 (m, 1H), 3.51 (t, J=6.6, 2H), 3.36-3.32 (m, 2H), 3.08-3.04 (m, 2H), 3.02-2.97 (m, 2H), 2.97-2.86 (m, 2H), 2.72 (s, 4H), 2.62-2.49 (m, 2H), 2.23-2.13 (m, 1H), 2.12-2.05 (m, 1H), 2.03-1.97 (m, 2H), 1.04 (d, J=6.4, 3H).
A. (4-[5-[5-Carbamoyl—(3-hydroxy-propyl)-4,5,6,7-tetrahydro-1 H-pyrazolo[4,3-c]pyridin-3-yl]-2-chloro-phenylethynyl]-benzyl)-(4-chloro-benzyl)-carbamic acid tert-butyl ester. This compound was prepared according to the methods described in Example 150, following sequentially Steps A, E, F, D, B, and then C, with the appropriate substituent changes. MS (ESI): mass calcd. for C37H39Cl2N5O4, 687.24; m/z found, 688.2 [M+H]+.
B. 3-[4-Chloro-3-[(4-{[(4-chlorobenzyl)amino]methyl]phenyl)ethynyl]phenyl}-1-{3-[(3aR,6aS)-tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl]propyl-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxamide. A solution of the carbamate above in 2.5:1 CH2Cl2:TFA (7 mL) was stirred at rt for 2 h. The reaction was quenched with satd. aq. NaHCO3 and extracted with CH2Cl2 (2×). The combined organic layers were dried and concentrated, and the residue was purified using preparatory HPLC (Method E; HCl salt). MS (ESI): mass calcd. for C38H40Cl2N6O2, 682.26; m/z found, 683.2 [M+H]+. 1H NMR: 7.91 (d, J=1.7, 1H), 7.71-7.62 (m, 3H), 7.60-7.55 (m, 3H), 7.56-7.45 (m, 4H), 4.67 (s, 2H), 4.29 (d, J=10.3, 4H), 4.26-4.21 (m, 2H), 4.00-3.88 (m, 1H), 3.86-3.77 (m, 4H), 3.72-3.65 (m, 1H), 3.59 (d, J=11.6, 1H), 3.53 (dd, J=9.6, 5.3, 1H), 3.34 (s, 3H), 3.25-3.15 (m, 2H), 3.14-3.05 (m, 1H), 2.86 (t, J=5.6, 2H), 2.77 (dd, J=10.6, 8.8, 1H), 2.40-2.23 (m, 2H).
A. 3-[4-Chloro-3-(3-formyl-phenylethynyl)-phenyl]-1-(3-thiomorpholin-4-yl-propyl)-1,4,6,7-tetrahydro-prazolo[4,3-c]pyridine-5-carboxylic acid amide. This compound was prepared according to the methods described in Example 150, following sequentially Steps A, B, C, E, and F, and then Example 167, Step C, with the appropriate substituent changes. MS (ESI): mass calcd. for C29H30ClN5O2S, 547.18; m/z found, 548.2 [M+H]+.
B. 3-(4-Chloro-3-{[3-(ff(4-chlorophenyl)methyl]amino]methyl)phenyl]ethynyl]phenyl)-1-(3-thiomorpholin-4-ylpropyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxamide. A solution of the aldehyde above (0.194 mmol), 4-chlorobenzylamine (0.388 mmol), and AcOH (16 μL) in CH2Cl2 (2 mL) was stirred for 30 min, and then treated with NaBH(OAc)3 (0.388 mmol). After 23 h, the mixture was diluted with satd. aq. NaHCO3 and extracted CH2Cl2 (3×). The combined organic layers were dried and concentrated. The residue was purified using preparatory HPLC (Method F; HCl salt). MS (ESI): mass calcd. for C36H38Cl2N6OS, 672.22; m/z found, 673.3 [M+H]+. 1H NMR: 7.84 (s, 1H), 7.70 (s, 1H), 7.58 (d, J=7.8, 2H), 7.53-7.35 (m, 7H), 4.60 (s, 2H), 4.21 (d, J=13.9, 4H), 4.16 (t, J=6.0, 2H), 3.77-3.68 (m, 4H), 3.21-3.07 (m, 5H), 3.03 (t, J=13.1, 2H), 2.79 (s, 2H), 2.74 (d, J=13.8, 2H), 2.32-2.25 (m, 2H).
The title compound was prepared using methods analogous to those described for Example 150, following sequentially Steps A, D, B, C, E, F, and then G. MS (ESI): mass calcd. for C43H44Cl2N6O2, 746.29; m/z found, 748.2 [M+H]+. 1H NMR: 7.65-7.51 (m, 4H), 7.41-7.39 (m, 1H), 7.25 (dd, J=8.6, 7.4, 1H), 6.92 (d, J=7.9, 1H), 4.34-4.11 (m, 4H), 3.56-2.92 (m, 24H).
A. 3-[4-Chloro-3-(4-formyl-phenylethynyl)-phenyl]-1-(3-thiomorpholin-4-yl-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid amide. The title compound was prepared using methods analogous to those described for Example 151. MS (ESI): mass calcd. for C29H30ClN5O2S, 548.10; m/z found 549.3 [M+H]+.
B. 3-(4-Chloro-3-{[4-(ff(4-methylphenyl)methyl]amino]methyl)phenyl]ethynyl]phenyl)-1-(3-thiomorpholin-4-ylpropyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxamide. The title compound was prepared using methods analogous to those described for Example 171, Step B. The compound was purified by Preparative HPLC Method B. MS (ESI): mass calcd. for C37H41ClN6OS, 652.29; m/z found, 653.3 [M+H]+. 1H NMR: 7.91 (d, J=2.1, 1H), 7.70-7.65 (m, 3H), 7.59 (dd, J=11.1, 8.4, 3H), 7.41 (d, J=8.1, 2H), 7.30 (d, J=7.9, 2H), 4.66 (s, 2H), 4.30-4.21 (m, 6H), 3.87-3.75 (m, 4H), 3.30-3.08 (m, 6H), 2.92-2.78 (m, 4H), 2.40-2.34 (m, 4H).
The compounds in Examples 174-176 were prepared using methods analogous to those described for Example 173.
MS (ESI): mass calcd. for C39H45ClN6O2S, 696.34; m/z found, 697.3 [M+H]+. 1H NMR: 7.92 (d, J=2.1, 1H), 7.71-7.67 (m, 3H), 7.62-7.54 (m, 3H), 7.47-7.35 (m, 2H), 7.01-6.98 (m, 2H), 4.72-4.62 (m, 3H), 4.29-4.19 (m, 5H), 3.87-3.70 (m, 3H), 3.30-3.15 (m, 4H), 3.14-2.75 (m, 6H), 2.38-2.32 (m, 2H), 1.33 (d, J=6.0, 6H).
MS (ESI): mass calcd. for C38H44ClN7OS, 681.33; m/z found, 682.3 [M+H]+. 1H NMR: 7.92 (d, J=2.1, 1H), 7.71-7.65 (m, 3H), 7.61-7.53 (m, 3H), 7.41-7.37 (m, 1H), 6.98 (d, J=8.8, 2H), 4.66 (s, 2H), 4.30-4.18 (m, 6H), 3.89-3.77 (m, 4H), 3.30-3.19 (m, 4H), 3.08-3.03 (m, 7H), 2.91-2.78 (m, 4H), 2.38-2.32 (m, 2H), 2.05 (s, 2H).
MS (ESI): mass calcd. for C37H39ClF2N6O2S, 704.27; m/z found, 705.2 [M+H]+. 1H NMR: 7.93 (d, J=2.1, 1H), 7.71-7.67 (m, 3H), 7.62-7.54 (m, 5H), 7.26 (d, J=8.6, 2H), 4.66 (s, 2H), 4.31 (d, J=11.1, 4H), 4.24 (t, J=6.5, 2H), 3.79 (t, J=5.7, 3H), 3.29-2.76 (m, 10H), 2.39-2.31 (m, 2H).
A. (4-Chlorobenzyl)-(4-ethynylbenzyl)-amine. Acetic acid (0.95 g, 15.7 mmol) was added at once to a solution of 4-chlorobenzylamine (2.2 g, 15.4 mmol) and 4-ethynylbenzaldehyde (1.0 g, 7.7 mmol) in CH2Cl2 (40 mL). The resulting slurry was stirred for 30 min and sodium triacetoxyborohydride (2.5 g, 11.6 mmol) was added portionwise. After 3 days, the reaction mixture was diluted with satd. aq. NaHCO3 and extracted with CH2Cl2 (×3). The combined organic extracts were dried (Na2SO4) and concentrated to give a yellow oil. Purification by silica gel chromatography (0 to 30% EtOAc/hexanes) afforded the title compound as a colorless oil (1.7 g, 85%). HPLC: Rt=4.70. MS (ESI): mass calcd. for C16H14ClN, 256.7; m/z found, 256.1 [M+H]+. 1H NMR (CDCl3): 7.46 (d, J=8.2, 2H), 7.31-7.27 (m, 4H), 3.78 (s, 2H), 3.75 (s, 2H), 3.06 (s, 1H), 1.58 (br s, 1H).
B. 1-[4-[(2-Chloro-5-[5-(methylsulfonyl)-1-{3-(4-pyridin-2-ylpiperidin-1-yl)propyl]-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-ylphenyl)ethynyl]phenyl]-N-[(4-chlorophenyl)methyl]methanamine. The title compound was prepared using methods analogous to those described for intermediate 1, substituting 2-(4 piperidinyl)pyridine for morpholine in Step D, and (4-chlorobenzyl)-(4-ethynylbenzyl)-amine for TMSA in Step E. HPLC: Rt=4.39. HPLC: Rt=4.39. MS (ESI): mass calcd. for C42H44Cl2N6O2S, 766.3; m/z found, 767.4 [M+H]+. 1H NMR (CDCl3): 8.55-8.52 (m, 1H), 7.82 (d, J=2.0, 1H), 7.62 (dt, J=7.7, 1.8, 1H), 7.56 (d, J=8.1, 2H), 7.49 (dd, J=8.4, 2.1, 1H), 7.45 (d, J=8.4, 1H), 7.38 (d, J=8.1, 2H), 7.30-7.27 (m, 4H), 7.17 (d, J=7.9, 1H), 7.12 (ddd, J=7.5, 4.9, 1.0, 1H), 4.54 (s, 2H), 4.12 (t, J=6.7, 2H), 3.81 (s, 2H), 3.76 (s, 2H), 3.66 (t, J=5.7, 2H), 3.00-2.91 (m, 5H), 2.89 (s, 3H), 2.74-2.65 (m, 1H), 2.33 (d, J=6.7, 2H), 2.14-2.02 (m, 4H), 1.98-1.91 (m, 2H), 1.87-1.75 (m, 2H).
The compounds in Examples 178-183 were prepared using methods analogous to those described for Example 177, substituting the appropriate amine for 2-(4-piperidinyl)pyridine in Step B.
HPLC: Rt=4.77. MS (ESI): mass calcd. for C43H45Cl2N5O2S, 765.3; m/z found, 766.3 [M+H]+. 1H NMR (CDCl3): 7.81 (d, J=2.0, 1H), 7.58-7.54 (m, 2H), 7.50 (dd, J=8.4, 2.1, 1H), 7.45 (d, J=8.4, 1H), 7.35-7.30 (m, 4H), 7.29 (d, J=2.7, 4H), 7.24-7.19 (m, 3H), 4.54 (s, 2H), 4.12 (t, J=6.8, 2H), 3.81 (s, 2H), 3.77 (s, 2H), 3.66 (t, J=5.7, 2H), 3.00-2.90 (m, 4H), 2.89 (s, 3H), 2.54-2.45 (m, 1H), 2.35 (t, J=6.9, 2H), 2.15-2.08 (m, 2H), 2.07-2.00 (m, 2H), 1.88-1.82 (m, 2H), 1.79-1.68 (m, 3H).
HPLC: Rt=4.35. MS (ESI): mass calcd. for C41H43Cl2N7O2S, 767.3; m/z found, 768.3 [M+H]+. 1H NMR (CDCl3): 8.20-8.17 (m, 1H), 7.81 (d, J=1.9, 1H), 7.59-7.55 (m, 2H), 7.57-7.44 (m, 3H), 7.36-7.33 (m, 2H), 7.30-7.28 (m, 4H), 6.66-6.61 (m, 2H), 4.54 (s, 2H), 4.13 (t, J=6.8, 2H), 3.81 (s, 2H), 3.77 (s, 2H), 3.65 (t, J=5.8, 2H), 3.55-3.51 (m, 4H), 2.93-2.91 (m, 2H), 2.90 (s, 3H), 2.54-2.51 (m, 4H), 2.37 (t, J=6.7, 2H), 2.16-2.09 (m, 2H), 1.64 (bs, 1H).
HPLC: Rt=4.76. MS (ESI): mass calcd. for C42H44Cl2N6O2S, 766.3; m/z found, 767.3 [M+H]+. 1H NMR (CDCl3): 7.81 (d, J=1.9, 1H), 7.58-7.55 (m, 2H), 7.49 (dd, J=8.4, 2.0, 1H), 7.45 (d, J=8.4, 1H), 7.36-7.32 (m, 2H), 7.30-7.25 (m, 6H), 6.94-6.90 (m, 2H), 6.88-6.83 (m, 1H), 4.53 (s, 2H), 4.12 (t, J=6.8, 2H), 3.80 (s, 2H), 3.76 (s, 2H), 3.64 (t, J=5.8, 2H), 3.20-3.17 (m, 4H), 2.92-2.89 (m, 2H), 2.89 (s, 3H), 2.58-2.55 (m, 4H), 2.37 (t, J=6.8, 2H), 2.15-2.08 (m, 2H), 1.87-1.83 (m, 1H).
HPLC: Rt=4.60. MS (ESI): mass calcd. for C37H41Cl2N5O2S, 689.2; m/z found, 690.3 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=1.9, 1H), 7.58-7.55 (m, 2H), 7.49 (dd, J=8.4, 2.0, 1H), 7.45 (d, J=8.4, 1H), 7.35-7.32 (m, 2H), 7.30-7.28 (m, 4H), 4.53 (s, 2H), 4.08 (t, J=6.8, 2H), 3.81 (2, 2H), 3.77 (s, 2H), 3.65 (t, J=5.8, 2H), 2.93-2.90 (m, 2H), 2.89 (s, 3H), 2.36-2.30 (bm, 4H), 2.25 (t, J=6.8, 2H), 2.10-2.02 (m, 2H), 1.83 (bs, 1H), 1.59-1.53 (m, 4H), 1.47-1.40 (m, 2H).
HPLC: Rt=4.53. MS (ESI): mass calcd. for C37H41Cl2N5O3S, 705.2; m/z found, 706.3 [M+H]+. 1H NMR (CDCl3): 7.80-7.79 (m, 1H), 7.56 (d, J=7.9, 2H), 7.48 (dd, J=8.4, 2.0, 1H), 7.45 (d, J=8.4, 1H), 7.34 (d, J=8.0, 2H), 7.31-7.27 (m, 4H), 4.53 (s, 2H), 4.12-4.01 (m, 2H), 3.81 (s, 2H), 3.81-3.79 (m, 1H), 3.79 (s, 2H), 3.68-3.61 (m, 4H), 3.24 (dd, J=11.0, 8.8, 1H), 2.95-2.85 (m, 2H), 2.90 (s, 3H), 2.80-2.70 (m, 2H), 2.44-2.36 (m, 1H), 2.30-2.20 (m, 2H), 2.12-1.98 (m, 2H), 1.77 (bs, 1H), 0.91 (d, J=6.3, 3H).
HPLC: Rt=4.57. MS (ESI): mass calcd. for C36H39Cl2N5O2S, 707.2; m/z found, 708.3 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=1.8, 1H), 7.56 (d, J=8.1, 2H), 7.48 (dd, J=8.4, 2.0, 1H), 7.45 (d, J=8.3, 1H), 7.34 (d, J=8.1, 2H), 7.30-7.28 (m, 6H), 4.53 (s, 2H), 4.07 (t, J=6.8, 2H), 3.81 (s, 2H), 3.79 (s, 2H), 3.64 (t, J=5.8, 2H), 3.28 (s, 3H), 2.89-2.85 (m, 2H), 2.68-2.63 (m, 8H), 2.34 (t, J=6.8, 2H), 2.09-2.01 (m, 2H).
A solution of DMF (15 mL) and distilled water (1.6 mL) was degassed with nitrogen for 1 h and transferred via syringe to a degassed flask containing 3-(4-chloro-3-iodophenyl)-5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine (2.3 g, 4.0 mmol), Pd(PPh3)2Cl2 (281 mg, 0.4 mmol), and CuI (76 mg, 0.4 mmol). The mixture was treated with 3-trimethylsilanylethynyl-benzaldehyde (1.2 g, 6.0 mmol) and DBU (1.8 g, 12 mmol), and the resulting mixture was stirred at rt under N2 for 18 h. The mixture was concentrated under high vacuum, and the crude residue was diluted with satd. aq. NaCl and extracted with CH2Cl2. The combined organic layers were dried and concentrated to give a brown oil. Purification by silica gel chromatography (0 to 3% MeOH/CH2Cl2) afforded the title compound as a beige foam (1.7 g, 76%). HPLC: Rt=4.86. MS (ESI): mass calcd. for C29H31ClN4O4S, 566.2; m/z found, 567.1 [M+H]+. 1H NMR (CDCl3): 10.00 (s, 1H), 8.10-8.09 (m, 1H), 7.90-7.87 (m, 1H), 7.86-7.84 (m, 1H), 7.82 (d, J=1.9, 1H), 7.57 (t, J=7.7, 1H), 7.52 (d, J=2.1, 1H), 7.48 (d, J=8.4, 1H), 4.55 (s, 2H), 4.11 (t, J=6.8, 2H), 3.70 (t, J=4.6, 4H), 3.67 (t, J=5.8, 2H), 2.93 (s, 3H), 2.93-2.89 (m, 2H), 2.44-2.40 (m, 4H), 2.34 (t, J=6.8, 2H), 2.12-2.06 (m, 2H).
To a solution of 3-({2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenyl}ethynyl)benzaldehyde (77 mg, 0.14 mmol) in CH2Cl2 (1.4 mL) were added 1-tetrahydro-2H-pyran-4-ylmethanamine (31 mg, 0.27 mmol) and ACOH (4 μL). The resulting slurry was stirred at rt for 30 min, then was treated with Na(OAc)3BH (57 mg, 0.27 mmol) and stirred for an additional 16 h. The mixture was diluted with satd. aq. NaHCO3 and extracted with CH2Cl2 (3×). The combined organic layers were dried and concentrated to afford a yellow oil. Purification of the residue (SiO2; 0-3% 2 M NH3 in MeOH/CH2Cl2) afforded the title compound as a white powder (72 mg, 79%). HPLC: Rt=4.24. MS (ESI): mass calcd. for C35H44ClN5O4S, 665.3; m/z found, 666.2 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=1.8, 1H), 7.68 (s, 1H), 7.51-7.47 (m, 2H), 7.46 (d, J=8.4, 1H), 7.33 (d, J=4.9, 1H), 4.54 (s, 2H), 4.10 (t, J=6.8, 2H), 4.02-3.95 (m, 3H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 3.43-3.35 (m, 3H), 2.91 (s, 3H), 2.91-2.88 (m, 2H), 2.43-2.38 (m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.04 (m, 2H), 1.77-1.61 (m, 4H), 1.37-1.25 (m, 4H).
The title compound was prepared from 1-[3-({2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenyl}ethynyl)phenyl]-N-(tetrahydro-2H-pyran-4-ylmethyl)methanamine using a method analogous to described for Example 2. HPLC: Rt=4.24. MS (ESI): mass calcd. for C35H48ClN5O4S, 669.3; m/z found, 670.3 [M+H]+. 1H NMR (CDCl3): 7.51 (d, J=2.1, 1H), 7.40 (d, J=8.3, 1H), 7.32 (dd, J=8.3, 2.1, 1H), 7.27 (t, J=7.5, 1H), 7.21-7.14 (m, 3H), 4.47 (s, 2H), 4.10 (t, J=6.8, 2H), 3.97 (dd, J=10.8, 3.5, 2H), 3.77 (s, 2H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 3.39 (dt, J=11.8, 1.8, 2H), 3.08-3.03 (m, 2H), 2.97-2.92 (m, 2H), 2.89 (s, 3H), 2.91-2.88 (m, 2H), 2.53 (d, J=6.5, 2H), 2.43-2.39 (m, 4H), 2.33 (t, J=6.8, 2H), 2.11-2.04 (m, 2H), 1.76-1.63 (m, 4H), 1.36-1.25 (m, 2H).
The compounds in Examples 187-188 were prepared using methods analogous to those described for Example 185.
HPLC: Rt=4.27. MS (ESI): mass calcd. for C35H39ClN6O3S, 658.3; m/z found, 659.2 [M+H]+. 1H NMR (CDCl3): 8.57 (d, J=4.4, 1H), 7.79 (d, J=1.9, 1H), 7.65 (dt, J=7.7, 1.8, 1H), 7.61 (s, 1H), 7.56-7.44 (m, 3H), 7.40-7.36 (m, 1H), 7.34 (t, J=7.4, 2H), 7.19-7.15 (m, 1H), 4.53 (s, 2H), 4.10 (t, J=6.8, 2H), 3.94 (s, 2H), 3.87 (s, 2H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 2.90 (s, 3H), 2.90-2.87 (m, 2H), 2.42-2.38 (m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.05 (m, 3H).
HPLC: Rt=4.42. MS (ESI): mass calcd. for C32H38ClN5O5S, 639.2; m/z found, 640.2 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=1.8, 1H), 7.58 (s, 1H), 7.52-7.47 (m, 2H), 7.45 (d, J=8.3, 1H), 7.35-7.33 (m, 2H), 4.53 (s, 2H), 4.10 (t, J=6.8, 2H), 3.83 (s, 2H), 3.74 (s, 3H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 3.44 (s, 2H), 2.91 (s, 3H), 2.90-2.87 (m, 2H), 2.43-2.38 (m, 4H), 2.33 (t, J=6.8, 2H), 2.11-2.04 (m, 2H), 1.96 (bs, 1H).
To a solution of methyl N-{[3-({2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenyl}ethynyl)phenyl]methyl}glycinate (46 mg, 0.07 mmol) in THF (0.3 mL) was added 1 N NaOH (0.3 mL). After 2 h at rt, the mixture was neutralized with 1 N HCl and extracted with 20% IPA/CHCl3 (3×). The combined organic extracts were dried and concentrated to give the title compound as a beige solid (36 mg, 82%). HPLC: Rt=4.17. MS (ESI): mass calcd. for C31H36ClN5O5S, 625.2; m/z found, 626.2 [M+H]+. 1H NMR (CDCl3): 7.74 (d, J=1.6, 1H), 7.64 (bs, 1H), 7.57 (m, 1H), 7.50 (dd, J=8.4, 1.9, 1H), 7.46 (d, J=8.4, 2H), 7.42-7.38 (bm, 1H), 4.51 (s, 2H), 4.09 (t, J=6.8, 4H), 3.73-3.68 (m, 6H), 3.64 (t, J=5.7, 2H), 3.43-3.39 (m, 2H), 2.92 (s, 3H), 2.91-2.87 (m, 2H), 2.45-2.41 (bm, 4H), 2.34 (t, J=7.0, 2H), 2.10-2.03 (m, 2H).
A. 7-Hydroxy-(3S)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid methyl ester. To a cooled (0° C.) slurry of 7-hydroxy-(3S)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (770 mg, 3.6 mmol) in MeOH (13 mL) was added oxalyl chloride (1.8 mL, 18 mmol) dropwise. The resulting mixture was heated at 65° C. for 5 h. After cooling to rt, the reaction was slowly quenched with satd. aq. NaHCO3, and the solvent was removed in vacuo. The crude residue was dissolved in 20% IPA/CHCl3 and washed with satd. aq NaHCO3. The organic layer was dried and concentrated to give the title compound as a white solid (523 mg, 63%). 1H NMR: 6.92 (d, J=8.3, 1H), 6.59 (dd, J=8.3, 2.5, 1H), 6.47 (d, J=2.4, 1H), 5.48 (s, 1H), 3.97 (d, J=15.9, 1H), 3.91 (d, J=15.9, 1H), 3.75 (s, 3H), 3.67 (dd, J=10.3, 4.7, 1H), 2.97 (dd, J=15.9, 4.7, 1H), 2.82 (dd, J=15.9, 10.3, 1H).
B. 7-Hydroxy-(3S)-3,4-dihydro-1H-isoquinoline-2,3-dicarboxylic acid 2-tert-butyl ester 3-methyl ester. To a solution of the above amine (100 mg, 0.48 mmol) in 2:1 THF:water (2.4 mL) was added di-tert-butyldicarbonate (105 mg, 0.48 mmol) dropwise. After stirring for 18 h, the reaction mixture was diluted with 0.5 N HCl and extracted with EtOAc (3×). The combined organic extracts were dried and concentrated to afford the title compound as a colorless oil (124 mg, 84%). 1H NMR (CDCl3): 7.21 (bs, 1H), 6.98 (d, J=8.0, 1H), 6.68 (dt, J=8.1, 2.3, 1H), 6.62-6.60 (m, 1H), 5.11-5.09 (m, 0.4H, minor rotamer), 4.74 (t, J=5.5, 0.6H, major rotamer), 4.68-4.59 (m, 1H), 4.50-4.40 (m, 1H), 3.66 (s, 1.8H, major rotamer), 3.59 (s, 1.2H, minor rotamer), 3.18-3.02 (m, 2H), 1.52 (s, 3.6H, minor rotamer), 1.46 (s, 5.4H, major rotamer).
C. 7-Trifluoromethanesulfonyloxy-(3S)-3,4-dihydro-1H-isoquinoline-2,3-dicarboxylic acid 2-tert-butyl ester 3-methyl ester. To an ice-cooled solution of the above phenol (157 mg, 0.51 mmol) in CH2Cl2 (2.6 mL) was added pyridine (161 mg, 2.04 mmol) and trifluoromethanesulfonic anhydride (288 mg, 1.02 mmol). The mixture was warmed to rt and stirred at rt for 4 h. The reaction mixture was diluted with 0.5 N HCl and extracted with CH2Cl2 (3×). The combined organic extracts were dried and concentrated to give the title compound as an orange oil (220 mg, 98%). 1H NMR (CDCl3): 7.23 (d, J=8.4, 1H), 7.11 (d, J=8.5, 1H), 7.06 (d, J=9.8, 1H), 5.20 (dd, J=6.2, 2.5, 0.4H, minor rotamer), 4.84 (t, J=5.2, 0.6H, major rotamer), 4.58-4.44 (m, 1H), 3.65 (s, 1.2H, minor rotamer), 3.64 (s, 1.8H, major rotamer), 3.30 (dd, J=16.2, 2.2, 0.6H, major rotamer), 3.20-3.16 (m, 1H), 3.16-3.12 (m, 0.4H, minor rotamer), 1.53 (s, 5.4H, major rotamer), 1.47 (s, 3.6H, minor rotamer).
D. 2-(1,1-Dimethylethyl) 3-methyl (3S)-7-([2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-phenylethynyl)-3,4-dihydroisoquinoline-2,3(1H)-dicarboxylate. The title compound was prepared using methods analogous to those described in Example 1, Step B, substituting DMF for THF and 7-trifluoromethanesulfonyloxy-(3S)-3,4-dihydro-1H-isoquinoline-2,3-dicarboxylic acid 2-tert-butyl ester 3-methyl ester for 5-iodoindole. HPLC: Rt=5.38. MS (ESI): mass calcd. for C38H46ClN5O7S, 751.3; m/z found, 752.1 [M+H]+. 1H NMR (CDCl3): 7.78-7.76 (m, 1H), 7.51-7.47 (m, 1H), 7.44 (d, J=8.4, 1H), 7.39 (d, J=8.7, 2H), 7.16 (d, J=7.7, 1H), 5.17 (dd, J=6.0, 2.8, 0.6H, major rotamer), 4.82 (t, J=5.2, 0.4H, minor rotamer), 4.74 (d, J=16.9, 1H), 4.56-4.47 (m, 1H), 4.53 (s, 2H), 4.10 (t, J=6.8, 2H), 3.69 (t, J=4.6, 4H), 3.67-3.63 (m, 2H), 3.65 (s, 1.2H, minor rotamer), 3.63 (s, 1.8H, major rotamer), 3.28 (dd, J=16.1, 2.6, 0.6H, major rotamer), 3.21-3.14 (m, 0.4H, minor rotamer), 3.19-3.17 (m, 1H), 2.90 (s, 3H), 2.90-2.86 (m, 2H), 2.42-2.38 (m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.03 (m, 2H), 1.54 (s, 5H, major rotamer), 1.50 (s, 4H, minor rotamer).
To a solution of 2-(1,1-dimethylethyl) 3-methyl (3S)-7-({2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenyl}ethynyl)-3,4-dihydroisoquinoline-2,3(1H)-dicarboxylate (379 mg, 0.50 mmol) in THF (2 mL) was added 1 N NaOH (2 mL). The reaction mixture was stirred at rt for 4 h and then neutralized with 1 N HCl. Following extraction with CH2Cl2 (2×) and 20% IPA/CHCl3, the combined organic extracts were dried and concentrated to afford the title compound as a white solid (356 mg, 96%). HPLC: Rt=5.14. MS (ESI): mass calcd. for C37H44ClN5O7S, 737.3; m/z found, 738.1 [M+H]+.
To a solution of (3S)-7-({2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenyl}ethynyl)-2-{[(1,1-dimethylethyl)oxy]carbonyl}-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (68 mg, 0.09 mmol), HOBt (14 mg, 0.10 mmol), and EDC (19 mg, 0.10 mmol) in CH2Cl2(1.0 mL) was added methylamine (2.0 M in THF; 70 μL, 0.14 mmol) and iPr2NEt (24 mg, 0.18 mmol). The reaction mixture was stirred at rt for 16 h, diluted with satd. aq. NaHCO3, and extracted with CH2Cl2 (3×). The combined organic extracts were dried and concentrated to give a brown oil. Purification (SiO2: 0 to 3% 2 M NH3 in MeOH/CH2Cl2) afforded a white foam, which was dissolved in 2:1 CH2Cl2:TFA (1.0 mL) and stirred at rt for 1 h. The mixture was diluted with satd. aq. NaHCO3 and extracted with CH2Cl2 (3×). The combined organic extracts were dried and concentrated to give the title compound as a white solid (34 mg, 56%). HPLC: Rt=4.12. MS (ESI): mass calcd. for C33H39ClN6O4S, 650.2; m/z found, 651.1 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.9, 1H), 7.49 (dd, J=8.4, 2.0, 1H), 7.45 (d, J=8.4, 1H), 7.40 (dd, J=7.8, 1.4, 1H), 7.30 (s, 1H), 7.23-7.20 (m, 1H), 7.17 (d, J=7.9, 1H), 4.53 (s, 2H), 4.10 (t, J=6.8, 2H), 4.00 (s, 2H), 3.79 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 3.55 (dd, J=10.5, 5.1, 1H), 3.27 (dd, J=16.9, 5.1, 1H), 2.91 (s, 3H), 2.91-2.88 (m, 3H), 2.86 (d, J=5.0, 3H), 2.85-2.80 (m, 1H), 2.43-2.39 (m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.05 (m, 2H), 1.80 (bs, 1H).
The compounds in Examples 193-194 were prepared using methods analogous to those described for Examples 190-192, substituting the appropriate amine for methylamine.
HPLC: Rt=4.17. MS (ESI): mass calcd. for C34H41ClN6O4S, 664.3; m/z found, 665.1 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.9, 1H), 7.49 (dd, J=8.4, 2.0, 1H), 7.45 (d, J=8.4, 1H), 7.37 (dd, J=7.9, 1.4, 1H), 7.30 (s, 1H), 7.10 (d, J=7.9, 1H), 4.54 (s, 2H), 4.13-4.08 (m, 4H), 3.94 (dd, J=10.8, 3.8, 1H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 3.14 (s, 3H), 3.02 (s, 3H), 2.99-2.94 (m, 1H), 2.91 (s, 3H), 2.91-2.88 (m, 2H), 2.85-2.79 (m, 1H), 2.43-2.39 (m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.04 (m, 2H), 1.94 (bs, 1H).
HPLC: Rt=4.10. MS (ESI): mass calcd. for C36H43ClN6O5S, 706.3; m/z found, 707.2 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.9, 1H), 7.49 (dd, J=8.4, 2.0, 1H), 7.45 (d, J=8.4, 1H), 7.38 (dd, J=7.9, 1.4, 1H), 7.29 (s, 1H), 7.11 (d, J=7.9, 1H), 4.53 (s, 2H), 4.12-4.08 (m, 4H), 3.89 (dd, J=10.9, 4.2, 1H), 3.76-3.72 (m, 4H), 3.71-3.68 (m, 7H), 3.65 (t, J=5.8, 2H), 3.56-3.52 (m, 1H), 3.07-2.99 (m, 1H), 2.90 (s, 3H), 2.90-2.87 (m, 2H), 2.79 (dd, J=16.9, 3.6, 1H), 2.43-2.38 (m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.04 (m, 2H), 1.90 (bs, 1H).
The title compound was prepared using methods analogous to those described for Examples 190-192, substituting ammonium chloride for methylamine, DMF for THF, and N-methylmorpholine for iPr2NEt in Example 192. HPLC: Rt=4.09. MS (ESI): mass calcd. for C32H37ClN6O4S, 636.2; m/z found, 637.1 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.9, 1H), 7.49 (dd, J=8.4, 2.0, 1H), 7.45 (d, J=8.4, 1H), 7.40 (dd, J=7.8, 1.4, 1H), 7.30 (s, 1H), 7.16 (d, J=7.9, 1H), 7.07 (bs, 1H), 5.73 (bs, 1H), 4.53 (s, 2H), 4.10 (t, J=6.8, 2H), 4.03 (s, 2H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 3.59 (dd, J=10.4, 5.1, 1H), 3.23 (dd, J=16.8, 5.1, 1H), 2.92-2.87 (m, 3H), 2.91 (s, 3H), 2.43-2.38 (m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.04 (m, 2H), 1.88 (bs, 1H).
The title compound was prepared using methods analogous to those described for Examples 190-192, substituting 7-hydroxy-(3R)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid for 7-hydroxy-(3S)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid in Example 190, Step A, and pyrrolidine for methylamine in Example 192. HPLC: Rt=4.24. MS (ESI): mass calcd. for C36H43ClN6O4S, 690.3; m/z found, 691.3 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.8, 1H), 7.49 (dd, J=8.4, 2.0, 1H), 7.45 (d, J=8.3, 1H), 7.37 (dd, J=7.9, 1.4, 1H), 7.29 (s, 1H), 7.10 (d, J=7.9, 1H), 4.54 (s, 2H), 4.13-4.08 (m, 4H), 3.77 (dd, J=11.0, 4.1, 1H), 3.72-3.68 (m, 5H), 3.65 (t, J=5.7, 2H), 3.58-3.44 (m, 3H), 3.00 (dd, J=16.6, 11.1, 1H), 2.91 (s, 3H), 2.91-2.87 (m, 2H), 2.83 (dd, J=16.8, 3.1, 1H), 2.43-2.39 (m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.05 (m, 2H), 2.02-1.87 (m, 6H).
The title compound was prepared using methods analogous to those described for Example 195, substituting 7-hydroxy-(3R)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid for 7-hydroxy-(3S)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid in Example 190, Step A. HPLC: Rt=4.10. MS (ESI): mass calcd. for C32H37ClN6O4S, 636.2; m/z found, 637.2 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.9, 1H), 7.49 (dd, J=8.4, 2.0, 1H), 7.45 (d, J=8.4, 1H), 7.40 (dd, J=7.8, 1.3, 1H), 7.30 (s, 1H), 7.16 (d, J=7.9, 1H), 7.06 (bs, 1H), 5.72 (bs, 1H), 4.53 (s, 2H), 4.10 (t, J=6.8, 2H), 4.03 (s, 2H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 3.59 (dd, J=10.4, 5.2, 1H), 3.23 (dd, J=16.9, 5.1, 1H), 2.93-2.85 (m, 3H), 2.90 (s, 3H), 2.43-2.38 (m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.04 (m, 2H).
To an ice-cooled solution of 2-(1,1-dimethylethyl) 3-methyl (3S)-7-({2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenyl}ethynyl)-3,4-dihydroisoquinoline-2,3(1H)-dicarboxylate (160 mg, 0.21 mmol) in CH2Cl2 was added TFA (0.8 mL). After 3 h, the mixture was diluted with satd. aq NaHCO3 and extracted with CH2Cl2 (3×). The combined organic extracts were dried and concentrated to afford the title compound as a white foam (133 mg, 96%). HPLC: Rt=4.22. MS (ESI): mass calcd. for C33H38ClN5O5S, 651.1; m/z found, 652.1 [M+H]+. 1H NMR (CDCl3): 7.77 (d, J=1.9, 1H), 7.48 (dd, J=8.4, 2.0, 1H), 7.44 (d, J=8.4, 1H), 7.37 (dd, J=7.8, 1.5, 1H), 7.29 (s, 1H), 7.12 (d, J=7.9, 1H), 4.52 (s, 2H), 4.13-4.07 (m, 4H), 3.78 (s, 3H), 3.78-3.74 (m, 1H), 3.69 (t, J=4.6, 4H), 3.64 (t, J=5.7, 2H), 3.11 (dd, J=16.6, 4.6, 1H), 3.02-2.94 (m, 1H), 2.90 (s, 3H), 2.90-2.86 (m, 2H), 2.43-2.38 (m, 4H), 2.32 (t, J=6.8, 2H), 2.13 (bs, 1H), 2.10-2.13 (m, 2H).
The title compound was prepared from (3S)-7-({2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenyl}ethynyl)-2-{[(1,1-di methylethyl)oxy]carbonyl}-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid using methods analogous to those described for Example 189. HPLC: Rt=4.10. MS (ESI): mass calcd. for C32H36ClN5O5S, 637.2; m/z found, 638.1 [M+H]+. 1H NMR: 7.86 (d, J=1.9, 1H), 7.60 (dd, J=8.5, 2.0, 1H), 7.56 (d, J=8.4, 1H), 7.55-7.49 (m, 3H), 7.37 (d, J=7.9, 1H), 4.55-4.47 (m, 1H), 4.51 (s, 2H), 4.45-4.40 (m, 1H), 4.23 (t, J=6.5, 2H), 4.06-4.00 (m, 2H), 3.76-3.68 (m, 2H), 3.65 (t, J=5.8, 2H), 3.56-3.47 (m, 3H), 3.33-3.29 (m, 3H), 3.28-3.22 (m, 2H), 3.17-3.09 (m, 2H), 2.98 (s, 3H), 2.92 (t, J=5.6, 2H), 2.38-2.30 (m, 2H).
The title compound was prepared from 2-(1,1-dimethylethyl) 3-methyl (3R)-7-({2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenyl}ethynyl)-3,4-dihydroisoquinoline-2,3(1H)-dicarboxylate using methods analogous to those described for Example 198. HPLC: Rt=4.23. MS (ESI): mass calcd. for C33H38ClN5O5S, 651.2; m/z found, 652.2 [M+H]+. 1H NMR (CDCl3): 7.77 (d, J=1.9, 1H), 7.48 (dd, J=8.4, 2.0, 1H), 7.44 (d, J=8.4, 1H), 7.37 (dd, J=7.9, 1.4, 1H), 7.29 (s, 1H), 7.12 (d, J=7.9, 1H), 4.52 (s, 2H), 4.14-4.08 (m, 4H), 3.79 (s, 3H), 3.78-3.74 (m, 1H), 3.69 (t, J=4.6, 4H), 3.64 (t, J=5.8, 2H), 3.11 (dd, J=16.6, 4.5, 1H), 2.98 (dd, J=16.6, 9.9, 1H), 2.90 (s, 3H), 2.90-2.87 (m, 2H), 2.42-2.38 (m, 4H), 2.32 (t, J=6.8, 2H), 2.20 (bs, 1H), 2.10-2.03 (m, 2H).
To a cooled (−78° C.) solution of methyl (3R)-7-({2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenyl}ethynyl)-1,2,3,4-tetrahydroisoquinoline-3-carboxylate (67 mg, 0.10 mmol) in THF was added DIBAL-H (1.5 M in toluene; 0.35 mL, 0.53 mmol). The reaction mixture was stirred at −78° C. for 1 h and at 0° C. for 30 min. After cooling to −78° C., the reaction was quenched with MeOH (1 mL), diluted with 1 N NaOH, and extracted with CH2Cl2 (3×). The combined organic extracts were dried and concentrated to give a brown oil. Purification (SiO2: 0 to 5% 2 M NH3 in MeOH/CH2Cl2) afforded the title compound as a yellow oil (19 mg, 30%). HPLC: Rt=4.13. MS (ESI): mass calcd. for C32H38ClN5O4S, 623.2; m/z found, 624.2 [M+H]+. 1H NMR (CDCl3): 7.77 (d, J=1.9, 1H), 7.48 (dd, J=8.4, 2.0, 1H), 7.44 (d, J=8.4, 1H), 7.39-7.35 (m, 1H), 7.29 (s, 1H), 7.09 (d, J=7.9, 1H), 4.53 (s, 2H), 4.10 (t, J=6.8, 2H), 4.07 (s, 2H), 3.82 (dd, J=10.8, 3.4, 1H), 3.70 (t, J=4.6, 4H), 3.65 (t, J=5.8, 2H), 3.55 (dd, J=10.6, 8.1, 1H), 3.15-3.08 (m, 1H), 2.91 (s, 3H), 2.91-2.87 (m, 2H), 2.75 (dd, J=16.7, 4.0, 1H), 2.61 (dd, J=16.7, 10.8, 1H), 2.43-2.38 (m, 4H), 2.33 (t, J=6.8, 2H), 2.21 (bs, 2H), 2.11-2.04 (m, 2H).
The title compound was prepared from (3R)-7-({2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenyl}ethynyl)-2-{[(1,1-di methylethyl)oxy]carbonyl}-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid using methods analogous to those described for Example 199. HPLC: Rt=4.16. MS (ESI): mass calcd. for C32H36ClN5O5S, 637.2; m/z found, 638.2 [M+H]+. 1H NMR: 7.85 (d, 1.8, 1H), 7.59 (dd, J=8.4, 2.0, 1H), 7.55 (d, J=8.4, 1H), 7.53-7.49 (m, 3H), 7.36 (d, J=8.0, 1H), 4.55-4.47 (m, 1H), 4.50 (s, 2H), 4.46-4.40 (m, 1H), 4.22 (t, J=6.5, 2H), 4.05-3.98 (m, 2H), 3.77-3.69 (m, 2H), 3.64 (t, J=5.7, 2H), 3.55-3.47 (m, 3H), 3.33-3.29 (m, 3H), 3.28-3.22 (m, 2H), 3.17-3.09 (m, 2H), 2.98 (s, 3H), 2.91 (t, J=5.6, 2H), 2.37-2.30 (m, 2H).
The title compound was prepared from 6-hydroxy-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid using methods analogous to those described for Example 190. HPLC: Rt=5.47. MS (ESI): mass calcd. for C32H38ClN5O4S, 751.3; m/z found, 752.1 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.9, 1H), 7.50 (dd, J=8.4, 2.1, 1H), 7.45 (d, J=8.4, 1H), 7.40 (s, 1H), 7.39-7.36 (m, 1H), 7.17 (d, J=7.9, 0.55H, major rotamer), 7.12 (d, J=7.9, 0.45H, minor rotamer), 5.19 (dd, J=6.1, 2.7, 0.55H, major rotamer), 4.84 (t, J=5.2, 0.45H, minor rotamer), 4.74 (dd, J=17.0, 7.5, 1H), 4.59-4.49 (m, 1H), 4.54 (s, 2H), 4.10 (t, J=6.8, 2H), 3.70 (t, J=4.6, 4H), 3.68-3.64 (m, 2H), 3.65 (s, 1.65H, major rotamer), 3.64 (s, 1.35H, minor rotamer), 3.29 (dd, J=16.0, 2.5, 0.55H, major rotamer), 3.18 (d, J=5.3, 1H), 3.15 (d, J=6.3, 0.45H, minor rotamer), 2.91 (s, 3H), 2.91-2.88 (m, 2H), 2.43-2.39 (m, 4H), 2.33 (t, J=6.8, 2H), 2.11-2.04 (m, 2H), 1.54 (s, 5H, major rotamer), 1.47 (s, 4H, minor rotamer).
The title compound was prepared from 2-(1,1-dimethylethyl) 3-methyl 6-({2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenyl}ethynyl)-3,4-dihydroisoquinoline-2,3(1H)-dicarboxylate using methods analogous to those described for Examples 191-192. HPLC: Rt=5.47. MS (ESI): mass calcd. for C37H45ClN6O4S, 704.3; m/z found, 705.2 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.9, 1H), 7.50 (dd, J=8.4, 2.1, 1H), 7.45 (d, J=8.3, 1H), 7.37 (d, J=7.9, 1H), 7.35 (s, 1H), 7.05 (d, J=7.9, 1H), 4.54 (s, 2H), 4.15-4.12 (m, 2H), 4.10 (t, J=6.9, 2H), 3.95 (dd, J=10.9, 4.2, 1H), 3.70 (t, J=4.6, 4H), 3.66 (t, J=5.8, 2H), 3.64-3.60 (m, 2H), 3.52-3.48 (m, 2H), 2.97-2.92 (m, 1H), 2.91 (s, 3H), 2.91-2.88 (m, 2H), 2.80 (dd, J=16.5, 3.7, 1H), 2.43-2.39 (m, 4H), 2.33 (t, J=6.8, 2H), 2.11-2.04 (m, 2H), 1.83 (bs, 1H), 1.71-1.57 (m, 6H).
The title compound was prepared from 2-(1,1-dimethylethyl) 3-methyl 6-({2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenyl}ethynyl)-3,4-dihydroisoquinoline-2,3(1H)-dicarboxylate using methods analogous to those described for Example 198. HPLC: Rt=4.17. MS (ESI): mass calcd. for C33H38ClN5O5S, 651.2; m/z found, 652.1 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=1.8, 1H), 7.49 (dd, J=8.4, 2.0, 1H), 7.45 (d, J=8.4, 1H), 7.39-7.36 (m, 2H), 7.04 (d, J=8.0, 1H), 4.53 (s, 2H), 4.16-4.08 (m, 4H), 3.79 (s, 3H), 3.78-3.74 (m, 2H), 3.70 (t, J=4.6, 4H), 3.66 (t, J=5.8, 2H), 3.11 (dd, J=16.4, 4.5, 1H), 2.98 (dd, J=16.3, 9.9, 1H), 2.91 (s, 3H), 2.91-2.88 (m, 2H), 2.43-2.38 (m, 4H), 2.33 (t, J=6.8, 2H), 2.11-2.04 (m, 2H).
The title compound was prepared from methyl 6-({2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenyl}ethynyl)-1,2,3,4-tetrahydroisoquinoline-3-carboxylate using methods analogous to those described for Example 201. HPLC: Rt=4.10. MS (ESI): mass calcd. for C32H38ClN5O4S, 623.2; m/z found, 624.2 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.8, 1H), 7.49 (dd, J=8.4, 2.0, 1H), 7.45 (d, J=8.4, 1H), 7.37 (d, J=8.1, 1H), 7.37 (s, 1H), 7.05 (d, J=7.8, 1H), 5.80 (s, 2H), 4.13-4.07 (m, 4H), 3.82 (dd, J=10.7, 3.5, 1H), 3.70 (t, J=4.6, 4H), 3.66 (t, J=5.8, 2H), 3.57-3.52 (m, 1H), 3.27-3.21 (m, 1H), 3.15-3.08 (m, 1H), 2.91 (s, 3H), 2.91-2.87 (m, 2H), 2.75 (dd, J=16.2, 3.8, 1H), 2.57 (dd, J=16.0, 10.9, 1H), 2.44-2.38 (m, 4H), 2.33 (t, J=6.8, 2H), 2.11-2.04 (m, 2H), 1.68-1.60 (bm, 1H).
The title compound was prepared from 6-hydroxy-1,2,3,4-tetrahydro-isoquinoline-1-carboxylic acid using methods analogous to those described for Example 190. HPLC: Rt=5.73. MS (ESI): mass calcd. for C38H46ClN5O7S, 751.3; m/z found, 752.3 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=2.0, 1H), 7.50 (dd, J=8.4, 2.0, 1H), 7.50-7.44 (m, 2H), 7.45 (d, J=8.4, 1H), 7.41 (s, 1H), 5.62 (s, 0.4H, minor rotamer), 5.46 (s, 0.6H, major rotamer), 4.54 (s, 2H), 4.13-4.08 (m, 2H), 3.81-3.75 (m, 2H), 3.73 (s, 3H), 3.70 (t, J=4.6, 4H), 3.66 (t, J=5.8, 2H), 2.98-2.93 (m, 2H), 2.91 (s, 3H), 2.91-2.87 (m, 2H), 2.43-2.39 (m, 4H), 2.33 (t, J=6.8, 2H), 2.11-2.04 (m, 2H), 1.51 (s, 3.6H, minor rotamer), 1.49 (5.4H, major rotamer).
The title compound was prepared from 2-(1,1-dimethylethyl) 1-methyl 6-({2-chloro-5-[5-(methylsulfonyl)-1-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenylethynyl)-3,4-dihydroisoquinoline-1,2(1H)-dicarboxylate using methods analogous to those described for Example 198. HPLC: Rt=4.23. MS (ESI): mass calcd. for C33H38ClN5O5S, 651.2; m/z found, 652.3 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.9, 1H), 7.49 (dd, J=8.4, 2.0, 1H), 7.45 (d, J=8.4, 1H), 7.40 (dd, J=7.9, 1.6, 1H), 7.38-7.35 (m, 2H), 4.76 (s, 1H), 4.54 (s, 2H), 4.10 (t, J=6.8, 2H), 3.78 (s, 3H), 3.70 (t, J=4.6, 4H), 3.66 (t, J=5.8, 2H), 3.32-3.25 (m, 1H), 3.09 (dt, J=12.6, 5.4, 1H), 2.91 (s, 3H), 2.91-2.88 (m, 2H), 2.87-2.75 (m, 2H), 2.43-2.39 (m, 4H), 2.33 (t, J=6.8, 2H), 2.11-2.04 (m, 1H).
The compounds in Examples 209-219 were prepared using methods analogous to those described for Example 177, substituting the appropriate amines for 4-chlorobenzylamine in Step A and 2-(4-piperidinyl)pyridine in Step B.
MS (ESI): mass calcd. for C37H42ClN5O4S, 687.3; m/z found, 688.3 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=1.6, 1H), 7.55 (d, J=8.2, 2H), 7.51-7.43 (m, 2H), 7.41-7.36 (m, 2H), 7.35-7.28 (m, 5H), 4.54 (s, 2H), 4.10 (t, J=6.8, 2H), 3.84-3.73 (m, 3H), 3.72-3.68 (m, 4H), 3.68-3.56 (m, 4H), 2.91 (s, 3H), 2.90-2.87 (m, 2H), 2.46-2.37 (m, 4H), 2.33 (t, J=6.8, 2H), 2.13-2.03 (m, 2H).
MS (ESI): mass calcd. for C37H42ClN5O3S, 671.3; m/z found, 672.3 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=1.7, 1H), 7.54 (d, J=8.2, 2H), 7.50-7.44 (m, 2H), 7.36 (d, J=4.4, 4H), 7.31-7.25 (m, 3H), 4.54 (s, 2H), 4.10 (t, J=6.8, 2H), 3.80 (q, J=6.6, 1H), 3.73-3.68 (m, 4H), 3.68-3.59 (m, 4H), 2.91 (s, 3H), 2.90-2.87 (m, 2H), 2.45-2.37 (m, 4H), 2.33 (t, J=6.8, 2H), 2.12-2.04 (m, 2H), 1.38 (d, J=6.6, 3H).
MS (ESI): mass calcd. for C38H42ClN5O5S, 715.3; m/z found, 716.3 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=1.8, 1H), 7.55 (d, J=8.2, 2H), 7.50-7.44 (m, 2H), 7.40-7.35 (m, 4H), 7.35-7.31 (m, 3H), 4.54 (s, 2H), 4.38 (s, 1H), 4.11 (t, J=6.8, 2H), 3.75 (s, 2H), 3.74-3.68 (m, 7H), 3.66 (t, J=5.8, 2H), 2.91 (s, 3H), 2.90-2.87 (m, 2H), 2.48-2.39 (m, 4H), 2.34 (t, J=6.6, 2H), 2.14-2.03 (m, 2H).
MS (ESI): mass calcd. for C37H40ClF3N6O2S, 724.3; m/z found, 725.2 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=1.8, 1H), 7.57 (d, J=8.2, 2H), 7.51-7.44 (m, 3H), 7.40 (d, J=8.2, 1H), 7.35 (d, J=8.2, 2H), 7.19 (dd, J=8.2, 1.9, 1H), 4.54 (s, 2H), 4.11 (t, J=6.8, 2H), 3.83-3.76 (m, 4H), 3.73-3.69 (m, 4H), 3.66 (t, J=5.7, 2H), 2.91 (s, 3H), 2.90-2.87 (m, 2H), 2.49-2.38 (m, 4H), 2.34 (t, J=6.8, 2H), 2.13-2.04 (m, 2H).
MS (ESI): mass calcd. for C43H46ClN7O3S, 775.3; m/z found, 776.2 [M+H]+. 1H NMR (CDCl3): 8.20-8.18 (m, 1H), 7.81 (d, J=1.9, 1H), 7.63-7.61 (m, 1H), 7.60-7.59 (m, 1H), 7.53-7.45 (m, 3H), 7.42 (d, J=8.2, 2H), 7.26-7.21 (m, 4H), 6.68-6.60 (m, 2H), 4.55 (s, 2H), 4.44-4.39 (m, 1H), 4.18-4.10 (m, 3H), 4.04 (d, J=2.4, 2H), 3.66 (t, J=5.8, 2H), 3.58-3.49 (m, 4H), 3.11-2.94 (m, 2H), 2.94-2.91 (m, 2H), 2.90 (s, 3H), 2.59-2.49 (m, 4H), 2.44-2.32 (m, 2H), 2.20-2.09 (m, 2H).
MS (ESI): mass calcd. for C44H47ClN6O3S, 774.3; m/z found, 775.2 [M+H]+. 1H NMR (CDCl3): 7.81 (d, J=1.9, 1H), 7.60 (d, J=8.2, 2H), 7.53-7.44 (m, 2H), 7.42 (d, J=8.2, 2H), 7.30-7.21 (m, 6H), 6.96-6.90 (m, 2H), 6.86 (t, J=7.3, 1H), 4.54 (s, 2H), 4.44-4.39 (m, 1H), 4.16-4.10 (m, 3H), 4.04 (d, J=2.5, 2H), 3.65 (t, J=5.8, 2H), 3.22-3.16 (m, 4H), 3.10-2.94 (m, 2H), 2.93-2.90 (m, 2H), 2.90 (s, 3H), 2.63-2.54 (m, 4H), 2.38 (t, J=6.8, 2H), 2.20-2.07 (m, 2H).
MS (ESI): mass calcd. for C44H48ClN7O2S, 773.3; m/z found, 774.2 [M+H]+. 1H NMR (CDCl3). 8.19 (dd, J=4.9, 1.3, 1H), 7.81 (d, J=1.9, 1H), 7.57 (d, J=8.2, 2H), 7.52-7.44 (m, 3H), 7.42 (d, J=8.2, 2H), 7.38-7.35 (m, 1H), 7.18-7.12 (m, 2H), 7.11-7.06 (m, 1H), 6.68-6.59 (m, 2H), 4.55 (s, 2H), 4.13 (t, J=6.8, 2H), 3.99-3.86 (m, 2H), 3.81 (t, J=4.9, 1H), 3.66 (t, J=5.7, 2H), 3.56-3.50 (m, 4H), 2.94-2.91 (m, 2H), 2.90 (s, 3H), 2.88-2.69 (m, 2H), 2.55-2.50 (m, 4H), 2.37 (t, J=6.7, 2H), 2.17-2.07 (m, 2H), 2.06-1.97 (m, 1H), 1.94-1.88 (m, 2H), 1.80-1.70 (m, 1H).
MS (ESI): mass calcd. for C44H48ClN7O2S, 773.3; m/z found, 774.2 [M+H]+. 1H NMR (CDCl3): 8.19 (dd, J=4.9, 1.2, 1H), 7.81 (d, J=1.8, 1H), 7.57 (d, J=8.2, 2H), 7.52-7.44 (m, 3H), 7.42 (d, J=8.3, 2H), 7.39-7.34 (m, 1H), 7.18-7.13 (m, 2H), 7.11-7.07 (m, 1H), 6.68-6.59 (m, 2H), 4.55 (s, 2H), 4.13 (t, J=6.8, 2H), 3.99-3.86 (m, 2H), 3.81 (t, J=4.9, 1H), 3.66 (t, J=5.8, 2H), 3.56-3.50 (m, 4H), 2.94-2.91 (m, 2H), 2.90 (s, 3H), 2.89-2.68 (m, 2H), 2.55-2.51 (m, 4H), 2.37 (t, J=6.8, 2H), 2.17-2.08 (m, 2H), 2.06-1.96 (m, 1H), 1.94-1.87 (m, 2H), 1.80-1.71 (m, 1H).
MS (ESI): mass calcd. for C45H49ClN6O2S, 772.3; m/z found, 773.3 [M+H]+. 1H NMR (CDCl3): 7.81 (d, J=1.9, 1H), 7.57 (d, J=8.2, 2H), 7.52-7.44 (m, 2H), 7.42 (d, J=8.2, 2H), 7.39-7.34 (m, 1H), 7.29-7.23 (m, 2H), 7.19-7.12 (m, 2H), 7.11-7.07 (m, 1H), 6.93 (d, J=7.9, 2H), 6.86 (t, J=7.3, 1H), 4.54 (s, 2H), 4.13 (t, J=6.8, 2H), 3.99-3.86 (m, 2H), 3.81 (t, J=4.9, 1H), 3.66 (t, J=5.7, 2H), 3.23-3.14 (m, 4H), 2.94-2.90 (m, 2H), 2.90 (s, 3H), 2.87-2.69 (m, 2H), 2.60-2.56 (m, 4H), 2.38 (t, J=6.8, 2H), 2.17-2.07 (m, 2H), 2.07-1.95 (m, 1H), 1.95-1.87 (m, 2H), 1.80-1.69 (m, 1H).
MS (ESI): mass calcd. for C42H46ClN7O3S, 763.3; m/z found, 764.2 [M+H]+. 1H NMR (CDCl3): 8.21-8.16 (m, 1H), 7.81 (d, J=1.8, 1H), 7.55 (d, J=8.2, 2H), 7.52-7.44 (m, 3H), 7.41-7.36 (m, 2H), 7.35-7.28 (m, 5H), 6.68-6.60 (m, 2H), 4.54 (s, 2H), 4.13 (t, J=6.8, 2H), 3.86-3.69 (m, 3H), 3.68-3.56 (m, 4H), 3.55-3.51 (m, 4H), 2.94-2.91 (m, 2H), 2.90 (s, 3H), 2.56-2.51 (m, 4H), 2.38 (t, J=6.8, 2H), 2.17-2.08 (m, 2H).
MS (ESI): mass calcd. for C42H46ClN7O2S, 747.3; m/z found, 748.2 [M+H]+. 1H NMR (CDCl3): 8.20-8.17 (m, 1H), 7.81 (d, J=1.8, 1H), 7.55 (d, J=8.2, 2H), 7.51-7.44 (m, 3H), 7.36 (d, J=4.4, 4H), 7.29 (d, J=8.2, 2H), 7.28-7.24 (m, 1H), 6.68-6.59 (m, 2H), 4.54 (s, 2H), 4.13 (t, J=6.8, 2H), 3.81 (q, J=6.6, 1H), 3.69-3.62 (m, 4H), 3.56-3.50 (m, 4H), 2.94-2.90 (m, 2H), 2.90 (s, 3H), 2.55-2.50 (m, 4H), 2.37 (t, J=6.8, 2H), 2.17-2.07 (m, 2H), 1.38 (d, J=6.6, 3H).
The title compound was prepared using methods analogous to those described for Example 285. MS (ESI): mass calcd. for C33H38Cl2N6O4, 652.2; m/z found, 653.2 [M+H]+. 1H NMR (CDCl3; mixture of rotamers): 7.78 (d, J=2.1, 1H), 7.65 (d, J=1.9, 1H), 7.56 (d, J=2.1, 0.4H), 7.54 (d, J=2.2, 0.6H), 7.46 (s, 0.6H), 7.45-7.43 (m, 0.8H), 7.42 (d, J=2.0, 0.6H), 7.37 (s, 0.6H), 7.35 (s, 0.4H), 4.69 (s, 2H), 4.59 (s, 2H), 4.16-3.99 (m, 2H), 3.92 (s, 2H), 3.84-3.75 (m, 3H), 3.74 (s, 3H), 3.69-3.59 (m, 2H), 3.48 (s, 2H), 3.24 (dd, J=11.1, 8.8, 1H), 2.88-2.69 (m, 4H), 2.47-2.35 (m, 1H), 2.30-2.20 (m, 2H), 2.13-1.96 (m, 3H), 0.91 (d, J=6.3, 3H).
A solution of methyl N-{[5-({5-[5-(aminocarbonyl)-1-{3-[(3S)-3-methylmorpholin-4-yl]propyl}-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]-2-chlorophenyl}ethynyl)-2-chlorophenyl]methyl}glycinate (387 mg, 0.592 mmol) and LiOH (71 mg, 2.96 mmol) in 3:1:1 THF:MeOH:H2O (3.2 mL) was stirred at rt for 18 h. The mixture was diluted with 1 M HCl and washed with CH2Cl2. The aqueous layer was concentrated and purified by reverse-phase HPLC (0-100% MeCN/5 mM NH4OH in H2O) to afford the desired product as a yellow solid (226 mg, 60%). MS (ESI): mass calcd. for C32H36Cl2N6O4, 638.2; m/z found, 639.1 [M+H]+. 1H NMR (CDCl3): 7.78 (s, 1H), 7.73-7.70 (m, 1H), 7.64-7.59 (m, 1H), 7.51-7.38 (m, 3H), 4.59 (s, 2H), 4.26 (s, 2H), 4.12-3.97 (m, 2H), 3.85-3.59 (m, 5H), 3.51 (s, 2H), 3.37 (td, J=3.3, 1.6, 1H), 3.30-3.22 (m, 1H), 2.84-2.70 (m, 4H), 2.51-2.40 (m, 1H), 2.36-2.25 (m, 2H), 2.15-1.92 (m, 2H), 0.94 (d, J=6.3, 3H).
The compounds in Examples 222-224 were prepared using methods analogous to those described for Example 285.
MS (ESI): mass calcd. for C33H40Cl2N6O3, 638.3; m/z found, 639.2 [M+H]+. 1H NMR (CDCl3): 7.77 (d, J=2.1, 1H), 7.59-7.54 (m, 2H), 7.47-7.41 (m, 2H), 7.36 (d, J=8.2, 1H), 4.78 (s, 2H), 4.60 (s, 2H), 4.15-3.98 (m, 2H), 3.90 (s, 2H), 3.85-3.81 (m, 2H), 3.81-3.71 (m, 3H), 3.69-3.58 (m, 2H), 3.24 (dd, J=11.1, 8.8, 1H), 2.92-2.88 (m, 2H), 2.86-2.69 (m, 5H), 2.47-2.34 (m, 1H), 2.30-2.19 (m, 2H), 2.12-1.96 (m, 2H), 1.80-1.72 (m, 2H), 0.90 (t, J=7.5, 3H).
MS (ESI): mass calcd. for C35H42Cl2N6O3, 664.3; m/z found, 665.2 [M+H]+. 1H NMR (CDCl3; mixture of rotamers): 7.78 (d, J=2.1, 1H), 7.66 (d, J=1.8, 1H), 7.56 (d, J=2.2, 0.4H), 7.54 (d, J=2.2, 0.6H), 7.47 (s, 0.6H), 7.45 (s, 0.4H), 7.42 (d, J=2.0, 0.4H), 7.40 (d, J=2.0, 0.6H), 7.36 (s, 0.6H), 7.34 (s, 0.4H), 4.67 (d, J=9.8, 3H), 4.17-4.01 (m, 3H), 3.95-3.91 (m, 2H), 3.90-3.82 (m, 1H), 3.81-3.72 (m, 3H), 3.69-3.58 (m, 2H), 3.28-3.19 (m, 1H), 2.85-2.65 (m, 6H), 2.44-2.33 (m, 1H), 2.31-2.20 (m, 2H), 2.12-1.95 (m, 3H), 1.93-1.85 (m, 2H), 1.64-1.53 (m, 1H), 0.91 (d, J=6.3, 3H).
MS (ESI): mass calcd. for C37H40Cl2N6O2, 670.3; m/z found, 671.2 [M+H]+. 1H NMR (CDCl3; mixture of rotamers): 7.79 (d, J=2.1, 1H), 7.66 (d, J=1.9, 1H), 7.55 (d, J=2.2, 0.4H), 7.53 (d, J=2.1, 0.6H), 7.47 (s, 0.6H), 7.45 (s, 0.4H), 7.43 (d, J=2.0, 0.4H), 7.41 (d, J=2.0, 0.6H), 7.40-7.32 (m, 5H), 7.30-7.24 (m, 1H), 4.65-4.59 (m, 3H), 4.15-3.99 (m, 2H), 3.92 (s, 2H), 3.84 (s, 2H), 3.82-3.71 (m, 3H), 3.69-3.58 (m, 2H), 3.24 (dd, J=11.1, 8.8, 1H), 2.89-2.67 (m, 4H), 2.48-2.34 (m, 1H), 2.31-2.19 (m, 2H), 2.12-1.96 (m, 2H), 0.91 (d, J=6.3, 3H).
The title compound was prepared using methods analogous to those described for Example 135. MS (ESI): mass calcd. for C37H41ClN6O2S, 668.3; m/z found, 669.3 [M+H]+. 1H NMR: 7.81-7.79 (m, 1H), 7.52-7.50 (m, 2H), 7.40-7.37 (m, 1H), 7.30 (s, 1H), 7.25 (dd, J=8.7, 7.4, 2H), 7.15 (d, J=7.9, 1H), 6.95 (d, J=7.9, 2H), 6.86 (t, J=7.3, 1H), 4.53 (s, 2H), 4.16 (t, J=6.7, 2H), 4.04 (s, 2H), 3.67 (t, J=5.8, 2H), 3.21-3.15 (m, 6H), 2.96 (s, 3H), 2.95-2.89 (m, 4H), 2.65-2.61 (m, 4H), 2.43 (t, J=7.2, 2H), 2.16-2.11 (m, 2H).
Formaldehyde (37% in H2O; 0.06 mL) and sodium triacetoxyborohydride (16 mg, 0.075 mmol) were added to a solution of 7-[(2-chloro-5-{5-(methylsulfonyl)-1-[3-(4-phenylpiperazin-1-yl)propyl]-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl}phenyl)ethynyl]-1,2,3,4-tetrahydroisoquinoline (25 mg, 0.037 mmol) in dichloroethane (0.1 mL) and the resulting mixture was stirred at rt for 18 h. Additional formaldehyde (37% in H2O, 0.06 mL) and sodium triacetoxyborohydride (16 mg, 0.075 mmol) were added and the mixture was stirred an additional 4 h. The mixture was diluted with 1 M NaOH and extracted with 20% isopropanol in CHCl3. The organic layer was washed with H2O, dried, and concentrated. Purification by preparative thin layer chromatography (SiO2; 3% 2 M NH3 in MeOH/CH2Cl2) afforded the desired product as a white solid (10 mg, 40%). MS (ESI): mass calcd. for C38H43ClN6O2S, 682.3; m/z found, 683.3 [M+H]+. 1H NMR (CDCl3): 7.78 (s, 1H), 7.51-7.48 (m, 2H), 7.42-7.38 (m, 1H), 7.33-7.24 (m, 3H), 7.16 (d, J=7.9, 1H), 6.95 (d, J=8.2, 2H), 6.89 (t, J=7.3, 1H), 4.54 (s, 2H), 4.14 (t, J=6.8, 2H), 3.74-3.70 (m, 2H), 3.67 (t, J=5.7, 2H), 3.26-3.18 (m, 4H), 3.01 (t, J=5.9, 2H), 2.97-2.91 (m, 5H), 2.85 (t, J=5.9, 2H), 2.68-2.62 (m, 4H), 2.55-2.52 (m, 3H), 2.45 (t, J=5.9, 2H), 2.21-2.08 (m, 2H).
The title compound was prepared using methods analogous to those described for Example 135. MS (ESI): mass calcd. for C34H41ClN6O3S, 648.3; m/z found, 649.3 [M+H]+. 1H NMR: 7.83 (s, 1H), 7.55 (s, 2H), 7.37-7.33 (m, 1H), 7.29 (s, 1H), 7.16 (d, J=8.0, 1H), 4.51 (s, 2H), 4.14 (t, J=6.5, 2H), 4.00 (s, 2H), 3.65 (t, J=5.8, 2H), 3.62-3.56 (m, 1H), 3.13 (t, J=6.0, 2H), 2.97 (s, 3H), 2.96-2.92 (m, 2H), 2.91-2.83 (m, 4H), 2.36 (t, J=7.2, 2H), 2.13-1.99 (m, 4H), 1.90 (s, 3H), 1.86-1.78 (m, 2H), 1.53-1.40 (m, 2H).
The compounds in Examples 227-228 were prepared using methods analogous to those described for Example 2.
MS (ESI): mass calcd. for C32H42ClN5O2S, 595.3; m/z found, 596.2 [M+H]+. 1H NMR: 7.44-7.39 (m, 3H), 7.07-7.04 (m, 2H), 6.92 (s, 1H), 4.40 (s, 2H), 4.12 (m, 2H), 4.05 (s, 2H), 3.64 (t, J=5.8, 2H), 3.21 (t, J=6.1, 2H), 3.10-3.02 (m, 2H), 2.97-2.84 (m, 9H), 2.55-2.42 (m, 4H), 2.41-2.36 (m, 2H), 2.12-2.01 (m, 2H), 1.66-1.56 (m, 4H), 1.53-1.42 (m, 2H).
MS (ESI): mass calcd. for C34H45ClN6O2S, 636.3; m/z found, 637.2 [M+H]+. 1H NMR (CDCl3): 7.58-7.49 (m, 1H), 7.42-7.37 (m, 1H), 7.34-7.29 (m, 1H), 7.09-7.00 (m, 2H), 6.97-6.86 (m, 1H), 4.48 (s, 2H), 4.09 (t, J=6.8, 2H), 4.00 (s, 2H), 3.66 (t, J=5.8, 2H), 3.13 (t, J=5.9, 2H), 3.06-3.00 (m, 2H), 2.93-2.84 (m, 8H), 2.77 (t, J=5.7, 2H), 2.70-2.55 (m, 4H), 2.47-2.35 (m, 3H), 2.30 (t, J=6.8, 3H), 2.12-1.99 (m, 2H), 0.53-0.35 (m, 4H).
The compounds in Examples 230-237 were prepared according to the methods described for Example 249.
MS (ESI): mass calcd. for C39H49Cl2N5O5S, 769.3; m/z found, 770.2 [M+H]+. 1H NMR (CDCl3): 7.75 (d, J=1.8, 1H), 7.52-7.42 (m, 4H), 7.36 (d, J=7.4, 2H), 4.54 (dd, J=14.5, 6.4, 2H), 4.20-3.95 (m, 5H), 3.70-3.57 (m, 2H), 3.10-3.00 (m, 2H), 2.95-2.85 (m, 2H), 2.85 (s, 3H), 2.70-2.35 (m, 4H), 2.30 (t, J=10.8, 2H), 2.03 (t, J=11.5, 2H), 1.70-1.60 (m, 4H), 1.42 (s, 9H), 1.40-1.10 (m, 4H).
MS (ESI): mass calcd. for C37H54ClN5O5S, 715.3; m/z found, 716.3 [M+H]+. 1H NMR (CDCl3): 7.60 (d, J=1.7, 1H), 7.35-7.30 (m, 2H), 4.48 (q, J=14.4, 2H), 4.10-3.85 (m, 5H), 3.65-3.52 (m, 2H), 3.00-2.82 (m, 4H), 2.80 (s, 3H), 2.60-2.50 (m, 2H), 2.35-2.30 (m, 2H), 2.30 (d, J=6.5, 2H), 2.22-2.18 (m, 2H), 1.95-1.85 (m, 2H), 1.65-1.55 (m, 4H), 1.38 (s, 9H), 1.40-1.10 (m, 5H), 1.00 (d, J=6.7, 6H).
MS (ESI): mass calcd. for C40H52ClN5O5S, 749.3; m/z found, 750.2 [M+H]+. 1H NMR (CDCl3): 7.62 (s, 1H), 7.40-7.30 (m, 4H), 7.28 (t, J=7.5, 2H), 7.21-7.17 (m, 1H), 4.48 (q, J=14.4, 2H), 4.10-3.88 (m, 5H), 3.85 (s, 2H), 3.65-3.52 (m, 2H), 3.00-2.92 (m, 2H), 2.85-2.80 (m, 2H), 2.80 (s, 3H), 2.57-2.50 (m, 2H), 2.43-2.33 (m, 2H), 2.23-2.18 (m, 2H), 1.95-1.90 (m, 2H), 1.65-1.55 (m, 4H), 1.38 (s, 9H), 1.40-1.10 (m, 4H).
MS (ESI): mass calcd. for C39H56ClN5O5S, 741.4; m/z found, 742.3 [M+H]+. 1H NMR (CDCl3): 7.58 (s, 1H), 7.32 (s, 2H), 4.45 (q, J=14.4, 2H), 4.15-3.88 (m, 5H), 3.62-3.52 (m, 2H), 3.07 (q, J=7.3, 1H), 3.00-2.82 (m, 4H), 2.80 (s, 3H), 2.60-2.38 (m, 4H), 2.25 (t, J=11.4, 1H), 1.94 (t, J=11.4, 1H), 1.85-1.80 (m, 2H), 1.75-1.67 (m, 2H), 1.65-1.40 (m, 7H), 1.38 (s, 9H), 1.40-1.10 (m, 9H).
MS (ESI): mass calcd. for C38H49ClN6O5S, 736.3; m/z found, 737.2 [M+H]+. 1H NMR (CDCl3): 8.57 (d, J=4.2, 1H), 7.80 (d, J=2.1, 1H), 7.65 (dt, J=7.7, 1.8, 1H), 7.55 (dd, J=7.8, 1.0, 1H), 7.47 (dd, J=8.4, 2.1, 1H), 7.40 (d, J=8.4, 1H), 7.20 (m, 1H), 4.47 (q, J=14.4, 2H), 4.10-3.85 (m, 5H), 3.65-3.50 (m, 2H), 3.00-2.75 (m, 4H), 2.80 (s, 3H), 2.60-2.50 (m, 2H), 2.35-2.25 (m, 2H), 2.17 (t, J=11.5, 2H), 1.90-1.80 (m, 2H), 1.70-1.60 (m, 4H), 1.38 (s, 9H), 1.40-1.10 (m, 4H).
MS (ESI): mass calcd. for C38H49ClN6O5S, 736.3; m/z found, 737.2 [M+H]+. 1H NMR (CDCl3): 8.75 (s, 1H), 8.50 (d, J=4.3, 1H), 7.80 (d, J=8.7, 1H), 7.74 (s, 1H), 7.45-7.35 (m, 2H), 7.25-7.22 (m, 1H), 4.48 (q, J=14.4, 2H), 4.10-3.90 (m, 5H), 3.65-3.52 (m, 2H), 3.00-2.93 (m, 2H), 2.87-2.82 (m, 2H), 2.80 (s, 3H), 2.60-2.50 (m, 2H), 2.43-2.33 (m, 2H), 2.22-2.18 (m, 2H), 1.97-1.90 (m, 2H), 1.65-1.55 (m, 4H), 1.38 (s, 9H), 1.40-1.10 (m, 4H).
MS (ESI): mass calcd. for C38H57ClN6O5S, 744.4; m/z found, 745.3 [M+H]+. 1H NMR (CDCl3): 7.62 (d, J=1.9, 1H), 7.40-7.30 (m, 2H), 4.48 (q, J=14.4, 2H), 4.10-3.88 (m, 5H), 3.65 (s, 2H), 3.65-3.52 (m, 2H), 3.00-2.80 (m, 4H), 2.80 (s, 3H), 2.62 (q, J=7.2, 4H), 2.57-2.50 (m, 2H), 2.35-2.25 (m, 2H), 2.20-2.10 (m, 2H), 1.90-1.80 (m, 2H), 1.65-1.55 (m, 4H), 1.38 (s, 9H), 1.40-1.10 (m, 4H), 1.13 (t, J=7.2, 6H).
MS (ESI): mass calcd. for C39H48Cl2N6O4, 734.3; m/z found, 735.2 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=2.0, 1H), 7.55-7.40 (m, 4H), 7.36 (d, J=8.6, 2H), 5.05 (br s, 2H), 4.54 (q, J=14.5, 2H), 4.20-3.95 (m, 5H), 3.80-3.60 (m, 2H), 2.95-2.65 (m, 6H), 2.37 (d, J=6.6, 2H), 2.22-2.15 (m, 2H), 1.95-1.85 (m, 2H), 1.70-1.60 (m, 4H), 1.42 (s, 9H), 1.40-1.10 (m, 4H).
The compounds in Examples 238-239 were prepared according to the methods described for Example 132.
MS (ESI): mass calcd. for C34H40Cl2N6O2, 634.3; m/z found, 635.2 [M+H]+. 1H NMR (CDCl3): 7.72 (d, J=2.0, 1H), 7.45-7.35 (m, 4H), 7.24 (d, J=8.6, 2H), 4.80 (br s, 2H), 4.52 (br s, 2H), 4.10-3.85 (m, 5H), 3.75-3.55 (m, 2H), 3.15 (d, J=12, 2H), 2.90-2.50 (m, 6H), 2.28 (d, J=6.6, 2H), 2.10 (t, J=11.2, 1H), 1.82 (t, J=11.2, 1H), 1.70-1.50 (m, 4H), 1.40-1.10 (m, 4H).
MS (ESI): mass calcd. for C32H46ClN5O3S, 615.3; m/z found, 616.2 [M+H]+. 1H NMR: 7.62 (s, 1H), 7.40 (s, 2H), 4.40-4.35 (m, 2H), 4.12-3.05 (m, 2H), 3.65-3.50 (m, 4H), 3.35-3.30 (m, 2H), 3.17-3.08 (m, 2H), 3.00-2.80 (m, 4H), 2.85 (s, 3H), 2.30 (d, J=6.5, 2H), 2.18-1.80 (m, 4H), 1.95-1.85 (m, 2H), 1.65-1.55 (m, 4H), 1.40-1.30 (m, 4H), 1.00 (d, J=6.7, 6H).
The title compound was prepared by methods analogous to those described for Example 74, using formaldehyde (37 wt % in H2O; 3 equiv.) in dichloroethane. MS (ESI): mass calcd. for C35H43Cl2N5O3S, 683.3; m/z found, 684.2 [M+H]+. 1H NMR (CDCl3): 7.72 (d, J=1.8, 1H), 7.47-7.38 (m, 4H), 7.28 (d, J=8.7, 2H), 4.50 (dd, J=14.5, 6.4, 2H), 4.10-3.85 (m, 5H), 3.70-3.52 (m, 2H), 3.05-2.80 (m, 4H), 2.80 (s, 3H), 2.30-2.10 (m, 4H), 2.25 (s, 3H), 1.95-1.80 (m, 4H), 1.65-1.55 (m, 4H), 1.40-0.90 (m, 4H).
The title compound was prepared using methods analogous to those described for Example 66. MS (ESI): mass calcd. for C36H43Cl2N5O4S, 711.2; m/z found, 712.2 [M+H]+. 1H NMR (CDCl3): 7.70 (s, 1H), 7.47-7.40 (m, 4H), 7.30-7.25 (m, 2H), 4.52-4.42 (m, 2H), 4.35-4.25 (m, 1H), 4.15-4.10 (m, 1H), 4.05-3.95 (m, 1H), 3.80-3.70 (m, 1H), 3.65-3.55 (m, 2H), 3.30-3.10 (m, 2H), 2.95-2.85 (m, 3H), 2.80 (s, 3H), 2.70-2.60 (m, 2H), 2.40 (t, J=7.6, 2H), 2.30-1.70 (m, 4H), 2.00 (s, 3H), 1.75-1.40 (m, 4H), 1.30-1.00 (m, 4H).
The compounds in Examples 242-248 were prepared using methods analogous to those described for Example 2, using the appropriate alkynes as starting materials.
MS (ESI): mass calcd. for C39H52Cl2N6O4, 738.3; m/z found, 739.2 [M+H]+. 1H NMR (CDCl3): 7.35-7.10 (m, 5H), 7.07 (d, J=7.4, 2H), 4.55 (br s, 2H), 4.40 (br s, 2H), 4.10-3.85 (m, 5H), 3.65-3.50 (m, 2H), 3.05-2.75 (m, 8H), 2.60-2.50 (m, 2H), 2.35-2.30 (d, J=6.5, 2H), 2.15 (t, J=10.3, 1H), 1.90 (t, J=10.5, 1H), 1.65-1.52 (m, 4H), 1.38 (s, 9H), 1.40-1.10 (m, 6H).
MS (ESI): mass calcd. for C38H44Cl2N6O3, 702.3; m/z found, 703.2 [M+H]+. 1H NMR (CDCl3): 7.50-7.15 (m, 11H), 5.65 (br s, 2H), 4.52 (br s, 2H), 4.10-4.00 (m, 2H), 3.70-3.55 (m, 9H), 3.23 (dd, J=11.1, 8.8, 1H), 3.10-3.00 (m, 2H), 2.95-2.90 (m, 2H), 2.80-2.70 (m, 4H), 2.43-2.35 (m, 1H), 2.30-2.20 (m, 2H), 2.10-1.95 (m, 2H), 0.90 (d, J=6.3, 3H).
MS (ESI): mass calcd. for C37H44Cl2N6O2, 674.3; m/z found, 675.2 [M+H]+. 1H NMR (CDCl3): 7.45-7.15 (m, 11H), 4.65 (br s, 2H), 4.52 (br s, 2H), 4.10-4.00 (m, 2H), 3.70-3.55 (m, 9H), 3.23 (dd, J=11.1, 8.8, 1H), 3.10-3.00 (m, 2H), 2.95-2.90 (m, 2H), 2.80-2.70 (m, 4H), 2.43-2.35 (m, 1H), 2.30-2.20 (m, 2H), 2.10-1.95 (m, 2H), 0.90 (d, J=6.3, 3H).
MS (ESI): mass calcd. for C39H53Cl2N5O5S, 773.3; m/z found, 774.2 [M+H]+. 1H NMR (CDCl3): 7.32-7.12 (m, 5H), 7.07 (d, J=7.4, 2H), 4.54 (q, J=14.5, 2H), 4.10-3.85 (m, 5H), 3.65-3.50 (m, 2H), 3.05-2.75 (m, 10H), 2.80 (s, 3H), 2.60-2.50 (m, 2H), 2.32-2.20 (m, 2H), 2.15 (t, J=11.4, 1H), 1.85 (t, J=11.4, 1H), 1.65-1.52 (m, 4H), 1.48 (s, 9H), 1.40-1.10 (m, 4H).
MS (ESI): mass calcd. for C34H45Cl2N5O3S, 673.3; m/z found, 674.2 [M+H]+. 1H NMR (CDCl3): 7.35-7.12 (m, 5H), 7.07 (d, J=7.4, 2H), 4.38 (q, J=14.5, 2H), 4.10-3.85 (m, 5H), 3.65-3.50 (m, 2H), 3.05-2.75 (m, 10H), 2.80 (s, 3H), 2.60-2.50 (m, 2H), 2.32-2.20 (m, 2H), 2.12 (t, J=11.4, 1H), 1.85 (t, J=11.4, 1H), 1.65-1.52 (m, 4H), 1.40-1.10 (m, 4H).
MS (ESI): mass calcd. for C36H47Cl2N5O4S, 715.3; m/z found, 716.2 [M+H]+. 1H NMR (CDCl3): 7.32-7.15 (m, 5H), 7.05 (d, J=7.4, 2H), 4.60-4.50 (br s, 1H), 4.38 (q, J=14.5, 2H), 4.15-3.90 (m, 5H), 3.80-3.70 (m, 2H), 3.60-3.50 (m, 2H), 3.20-3.05 (m, 2H), 3.05-2.75 (m, 6H), 2.80 (s, 3H), 2.60-2.50 (m, 2H), 2.32-2.20 (m, 4H), 2.00 (s, 3H), 1.70-1.60 (m, 4H), 1.40-1.10 (m, 4H).
MS (ESI): mass calcd. for C35H47Cl2N5O3S, 687.3; m/z found, 688.2 [M+H]+. 1H NMR (CDCl3): 7.82-7.18 (m, 5H), 7.07 (d, J=7.4, 2H), 4.38 (q, J=14.5, 2H), 4.15-4.00 (m, 2H), 3.92-3.85 (m, 1H), 3.65-3.50 (m, 2H), 3.05-2.75 (m, 9H), 2.80 (s, 3H), 2.32-2.10 (m, 3H), 2.25 (s, 3H), 2.00-1.80 (m, 4H), 1.68-1.58 (m, 4H), 1.40-0.90 (m, 6H).
A. (R)-3-(4-Chloro-3-iodo-phenyl)-1-oxiranylmethyl-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester. (S)-(+)-Glycidyinosylate (17.5 g, 67.5 mmol) was added to a stirring mixture of 3-(4-chloro-3-iodo-phenyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester (15.1 g, 33.0 mmol) and Cs2CO3 (21.8 g, 67.0 mmol) in DMF (200 mL) at rt. After 21 h, the mixture was filtered and concentrated to give a crude oil. The oil was diluted with CH2Cl2 (150 mL) and washed with satd. aq. Na2CO3 (2×75 mL). The organic layer was dried and concentrated to give a gold oil. The oil was purified on SiO2 (CH2Cl2 to 5% acetone/CH2Cl2) to provide the desired product (11.9 g, 70%) as a white solid. TLC: Rf=0.36 [5% acetone/CH2Cl2]. MS (ESI): mass calcd. for C20H23CIIN3O3, 515.1; m/z found, 516.0 [M+H]+. 1H NMR (d6-DMSO): 8.07 (s, 1H), 7.55 (d, J=8.4, 1H), 7.47 (br s, 1H), 4.45 (s, 2H), 4.33 (dd, J=14.8, 3.2, 1H), 4.03 (dd, J=15.2, 6.0, 1H), 3.62-3.48 (m, 2H), 3.23 (br s, 1H), 2.74-2.68 (m, 1H), 2.65 (br s, 2H), 2.49-2.45 (m, 1H), 1.32 (s, 9H).
B. (S)-3-(4-Chloro-3-iodo-phenyl)-1-(2-hydroxy-3-thiomorpholin-4-yl-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]oyridine-5-carboxylic acid tert-butyl ester. Thiomorpholine (11 mL, 116 mmol) was added to a stirring mixture of (R)-3-(4-chloro-3-iodo-phenyl)-1-oxiranylmethyl-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester (11.8 g, 22.9 mmol) in EtOH (120 mL). After 20 h, the mixture was concentrated to give a golden oil. The oil was purified on SiO2 (CH2Cl2 to 20% acetone/CH2Cl2) to give the desired product (13.7 g, 97%) as a white solid. TLC: Rf=0.46 [15% acetone/CH2Cl2]. MS (ESI): mass calcd. for C24H32CIIN4O3S, 618.1; m/z found, 619.1 [M+H]+. 1H NMR (d6-DMSO): 7.97 (s, 1H), 7.46 (d, J=8.0, 1H), 7.38 (br s, 1H), 4.72 (d, J=4.8, 1H), 4.36 (s, 2H), 3.98-3.92 (m, 1H), 3.89-3.71 (m, 2H), 3.45 (br s, 2H), 2.58 (d, J=4.4, 2H), 2.51 (d, J=4.8, 4H), 2.42-2.35 (m, 4H), 2.18 (t, J=6.0, 2H), 1.24 (s, 9H).
C. 1-[3-(4-Chloro-3-iodo-phenyl)-4,5,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-1-yl-3-thiomorpholin-4-yl-propan-2-ol. Trifluoroacetic acid (30 mL) was added to a stirring solution of (S)-3-(4-chloro-3-iodo-phenyl)-1-(2-hydroxy-3-thiomorpholin-4-yl-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester (13.6 g, 22.0 mmol) in CH2Cl2 (30 mL). After 2 h, the mixture was concentrated to give a golden liquid. The liquid was set stirring in CH2Cl2 (200 mL) and satd. aq. K2CO3 (200 mL) was added slowly over 15 min. The organic layer was separated, dried, and concentrated to give the desired product (9.3 g, 82%) as a white solid. MS (ESI): mass calcd. for C19H24CIIN4OS, 518.0; m/z found, 519.0 [M+H]+. 1H NMR (d6-DMSO): 8.34 (s, 1H), 7.79 (d, J=8.4, 1H), 7.74 (dd, J=8.4, 2.0, 1H), 5.06 (d, J=4.8, 1H), 4.28 (dd, J=13.6, 3.6, 1H), 4.27-4.23 (m, 1H), 4.22-4.07 (m, 1H), 4.05 (s, 2H), 3.12 (br s, 2H), 2.95-2.73 (m, 11H), 2.54 (t, J=6.0, 2H).
D. (S)-2-[3-(4-Chloro-3-iodo-phenyl)-1-(2-hydroxy-3-thiomorpholin-4-yl-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-5-yl]-2-oxo-acetamide. Oxamic acid (1.76 g, 19.8 mmol) was added to a stirring mixture of 1,1′-carbonyldiimidazole (CDI) (3.19 g, 19.7 mmol) in DMF (50 mL). After 75 min, a solution of 1-[3-(4-chloro-3-iodo-phenyl)-4,5,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-1-yl]-3-thiomorpholin-4-yl-propan-2-ol (5.3 g, 10.2 mmol) in DMF (50 mL) was added and the mixture was stirred at rt. After 15 min, the mixture was concentrated to give a cloudy yellow liquid. The liquid was diluted with EtOAc (200 mL) and washed twice with H2O (200 mL). The organic layer was dried and concentrated to give the desired product (6.0 g, 100%) as a white solid. MS (ESI): mass calcd. for C21H25CIIN5O3S, 589.0; m/z found, 590.0 [M+H]+. 1H NMR (d6-DMSO): 8.30-8.25 (m, 2H), 8.21-7.90 (m, 2H), 7.88-7.50 (m, 1H), 5.06-4.75 (m, 3H), 4.28-4.00 (m, 2H), 3.97-3.78 (m, 2H), 3.05-2.75 (m, 11H), 2.62-2.42 (m, 2H).
E. (4-Chloro-benzyl)-(4-ethynyl-benzyl)-amine. 4-Trimethylsilanylethynyl-benzaldehyde (commercially available or available via Sonogashira coupling; 1.0 g, 4.9 mmol, 1.0 equiv) was dissolved in MeOH (20 mL) and 4-chlorobenzylamine (0.74 g, 5.2 mmol, 1.05 equiv) was added. After stirring at rt for 16 h, NaBH4 (0.19 g, 4.9 mmol, 1.0 equiv) was added slowly. 1 N HCl was slowly added to quench the unreacted NaBH4. The mixture was concentrated and the residue was partitioned between CH2Cl2 and satd. aq. NaHCO3. The organic layer was dried and concentrated. The crude compound was purified (SiO2; EtOAc/hexanes) to provide the title compound (1.1 g, 4.3 mmol, 90%). TLC: Rf=0.54 (50% EtOAc/hexanes). Alternatively, the material was converted to the corresponding HCl salt and purified by recrystallization.
F. (S)-2-[3-(4-Chloro-3-[4-[(4-chloro-benzylamino)-methyl]-phenylethynyl]-phenyl)-1-(2-hydroxy-3-thiomorpholin-4-yl-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-5-yl]-2-oxo-acetamide. Triethylamine (1.5 mL, 10.8 mmol) was added to a stirring mixture of (S)-2-[3-(4-chloro-3-iodo-phenyl)-1-(2-hydroxy-3-thiomorpholin-4-yl-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-5-yl]-2-oxo-acetamide (2.1 g, 3.5 mmol) and (4-chloro-benzyl)-(4-ethynyl-benzyl)-amine (1.5 g, 6.0 mmol) in DMF (15 mL). The flask was equipped with a nitrogen inlet, septum, and vacuum line. The mixture was evacuated and flushed with nitrogen three times. To the mixture was added Pd(PPh3)2Cl2 (51 mg, 0.073 mmol) and CuI (36 mg, 0.19 mmol) in one portion while keeping the contents under a positive pressure of nitrogen. After 3 h, the mixture was added to H2O (100 mL) and EtOAc (100 mL). The organic layer was separated, washed once with H2O (50 mL), dried, and concentrated to give a golden brown oil. The oil was purified on SiO2 (CH2Cl2 to 100% (5% MeOH(2 M NH3)/CH2Cl2)) to give the desired product (1.72 g, 68%) as a light yellow solid. TLC: Rf=0.42 [10% MeOH(2 M NH3)/CH2Cl2]. MS (ESI): mass calcd. for C37H38Cl2N6O3S, 716.2; m/z found, 717.2 [M+H]+. 1H NMR (d6-DMSO): 8.00-7.88 (m, 2H), 7.82-7.75 (m, 2H), 7.74-7.65 (m, 2H), 7.55 (d, J=8.4, 2H), 7.50 (s, 4H), 5.08-5.00 (m, 1H), 4.89-4.83 (m, 2H), 4.30-4.23 (m, 1H), 4.21-4.07 (m, 2H), 3.95-3.72 (m, 6H), 3.12-2.91 (m, 4H), 2.90-2.80 (m, 4H), 2.79-2.69 (m, 4H), 2.54-2.47 (m, 2H).
The compounds in Examples 250-275 were prepared using methods analogous to those described for Example 249, with exceptions as noted.
MS (ESI): mass calcd. for C37H38Cl2N6O3S, 716.2; m/z found, 717.2 [M+H]+. 1H NMR (d6-DMSO): 8.00-7.88 (m, 2H), 7.82-7.75 (m, 2H), 7.74-7.65 (m, 2H), 7.55 (d, J=8.4, 2H), 7.50 (s, 4H), 5.08-5.00 (m, 1H), 4.89-4.83 (m, 2H), 4.30-4.23 (m, 1H), 4.21-4.07 (m, 2H), 3.95-3.72 (m, 6H), 3.12-2.91 (m, 4H), 2.90-2.80 (m, 4H), 2.79-2.69 (m, 4H), 2.54-2.47 (m, 2H).
MS (ESI): mass calcd. for C36H38Cl2N6O2S, 688.2; m/z found, 689.2 [M+H]+. 1H NMR (d6-DMSO): 10.11 (s, 1H), 9.69 (s, 2H), 7.83 (s, 1H), 7.65-7.48 (m, 8H), 7.42 (d, J=8.4, 2H), 4.54-4.44 (m, 6H), 4.18-4.00 (m, 5H), 3.68-3.52 (m, 4H), 3.35-3.25 (m, 1H), 3.24-3.00 (m, 6H), 2.78-2.60 (m, 4H).
MS (ESI): mass calcd. for C36H38Cl2N6O2S, 688.2; m/z found, 689.2 [M+H]+. 1H NMR (d6-DMSO): 10.11 (s, 1H), 9.69 (s, 2H), 7.83 (s, 1H), 7.65-7.48 (m, 8H), 7.42 (d, J=8.4, 2H), 4.54-4.44 (m, 6H), 4.18-4.00 (m, 5H), 3.68-3.52 (m, 4H), 3.35-3.25 (m, 1H), 3.24-3.00 (m, 6H), 2.78-2.60 (m, 4H).
MS (ESI): mass calculated for C37H38Cl2N6O4, 700.2; m/z found, 701.2 [M+H]+. 1H NMR (d6-DMSO) 8.16 (s, 1H), 7.90-7.75 (m, 2H), 7.73-7.50 (m, 4H), 7.43 (d, J=8.4, 2H), 7.38 (s, 4H), 5.00-4.95 (m, 1H), 4.80-4.70 (m, 2H), 4.25-4.15 (m, 1H), 4.13-3.95 (m, 2H), 3.94-3.65 (m, 6H), 3.59-3.50 (br s, 4H), 2.90-2.80 (m, 3H), 2.48-2.30 (m, 6H).
MS (ESI): mass calcd. for C37H38Cl2N6O4, 700.2; m/z found, 701.2 [M+H]+. 1H NMR (d6-DMSO): 8.16 (s, 1H), 7.90-7.75 (m, 2H), 7.73-7.50 (m, 4H), 7.43 (d, J=8.4, 2H), 7.38 (s, 4H), 5.00-4.95 (m, 1H), 4.80-4.70 (m, 2H), 4.25-4.15 (m, 1H), 4.13-3.95 (m, 2H), 3.94-3.65 (m, 6H), 3.59-3.50 (br s, 4H), 2.90-2.80 (m, 3H), 2.48-2.30 (m, 6H).
MS (ESI): mass calcd. for C38H40Cl2N6O4, 714.3; m/z found, 715.2 [M+H]+. 1H NMR (d6-DMSO): 12.40 (br s, 1H), 10.91 (br s, 2H), 9.16 (s, 1H), 8.90-8.35 (m, 10H), 8.28 (d, J=9.8, 2H), 7.38 (s, 4H), 5.05-5.00 (m, 1H), 4.70-4.60 (m, 1H), 4.50-4.35 (m, 6H), 4.32-3.85 (m, 5H), 3.82-3.65 (m, 3H), 3.50-3.15 m, 2H), 3.10-2.90 (m, 2H), 1.06 (d, J=7.6, 2H).
MS (ESI): mass calcd. for C38H40Cl2N6O4, 714.3; m/z found, 715.2 [M+H]+. 1H NMR (d6-DMSO): 12.40 (br s, 1H), 10.91 (br s, 2H), 9.16 (s, 1H), 8.90-8.35 (m, 10H), 8.28 (d, J=9.8, 2H), 7.38 (s, 4H), 5.05-5.00 (m, 1H), 4.70-4.60 (m, 1H), 4.50-4.35 (m, 6H), 4.32-3.85 (m, 5H), 3.82-3.65 (m, 3H), 3.50-3.15 m, 2H), 3.10-2.90 (m, 2H), 1.06 (d, J=7.6, 2H).
MS (ESI): mass calcd. for C37H40Cl2N6O3, 686.3; m/z found, 687.2 [M+H]+. 1H NMR (d6-DMSO): 12.30 (br s, 1H), 10.83 (br s, 2H), 8.73 (s, 1H), 8.65-8.38 (m, 10H), 8.27 (d, J=7.6, 2H), 4.98-4.35 (m, 8H), 4.25-3.95 (m, 4H), 3.92-3.55 (m, 7H), 3.50-3.15 m, 3H), 2.79 (s, 2H), 1.05 (d, J=7.8, 2H).
MS (ESI): mass calcd. for C37H40Cl2N6O3, 686.3; m/z found, 687.2 [M+H]+. 1H NMR (d6-DMSO): 12.30 (br s, 1H), 10.83 (br s, 2H), 8.73 (s, 1H), 8.65-8.38 (m, 10H), 8.27 (d, J=7.6, 2H), 4.98-4.35 (m, 8H), 4.25-3.95 (m, 4H), 3.92-3.55 (m, 7H), 3.50-3.15 m, 3H), 2.79 (s, 2H), 1.05 (d, J=7.8, 2H).
MS (ESI): mass calcd. for C36H38Cl2N6O3, 672.2; m/z found, 673.2 [M+H]+. 1H NMR (d6-DMSO): 10.30 (br s, 1H), 9.77 (br s, 2H), 7.92 (s, 1H), 7.80-7.55 (m, 10H), 7.52 (d, J=6.6, 2H), 4.58 (s, 2H), 4.40 (br s, 1H), 4.25-3.90 (m, 9H), 3.89-3.73 (m, 2H), 3.70-3.58 (br s, 2H), 3.56-3.38 (m, 3H), 3.16 (br s, 3H), 2.77 (s, 2H).
MS (ESI): mass calcd. for C36H38Cl2N6O3, 672.2; m/z found, 673.2 [M+H]+. 1H NMR (d6-DMSO): 10.30 (br s, 1H), 9.77 (br s, 2H), 7.92 (s, 1H), 7.80-7.55 (m, 10H), 7.52 (d, J=6.6, 2H), 4.58 (s, 2H), 4.40 (br s, 1H), 4.25-3.90 (m, 9H), 3.89-3.73 (m, 2H), 3.70-3.58 (br s, 2H), 3.56-3.38 (m, 3H), 3.16 (br s, 3H), 2.77 (s, 2H).
MS (ESI): mass calcd. for C36H39Cl2N5O3S2, 723.2; m/z found, 724.2 [M+H]+. 1H NMR (d6-DMSO): 10.23 (br s, 1H), 9.78 (br s, 2H), 7.90 (s, 1H), 7.75-7.58 (m, 8H), 7.51 (d, J=8.4, 2H), 4.46 (s, 2H), 4.30-4.05 (m, 6H), 3.82-3.68 (m, 2H), 3.55-3.45 (m, 2H), 3.40-3.32 (m, 1H), 3.25-3.00 (m, 10H), 2.93 (br s, 2H), 2.86-2.76 (m, 2H).
MS (ESI): mass calcd. for C36H39Cl2N5O3S2, 723.2; m/z found, 724.2 [M+H]+. 1H NMR (d6-DMSO): 10.23 (br s, 1H), 9.78 (br s, 2H), 7.90 (s, 1H), 7.75-7.58 (m, 8H), 7.51 (d, J=8.4, 2H), 4.46 (s, 2H), 4.30-4.05 (m, 6H), 3.82-3.68 (m, 2H), 3.55-3.45 (m, 2H), 3.40-3.32 (m, 1H), 3.25-3.00 (m, 10H), 2.93 (br s, 2H), 2.86-2.76 (m, 2H).
MS (ESI): mass calcd. for C36H39Cl2N5O4S, 707.2; m/z found, 708.1 [M+H]+. 1H NMR (d6-DMSO): 10.28 (br s, 1H), 9.71 (br s, 2H), 7.91 (s, 1H), 7.74-7.57 (m, 8H), 7.52 (d, J=6.5, 2H), 4.47 (s, 3H), 4.28-4.06 (m, 6H), 3.98-3.90 (m, 2H), 3.89-3.27 (m, 10H), 3.23-3.05 (m, 4H), 2.94 (br s, 2H).
MS (ESI): mass calcd. for C36H39Cl2N5O4S, 707.2; m/z found, 708.1 [M+H]+. 1H NMR (d6-DMSO): 10.28 (br s, 1H), 9.71 (br s, 2H), 7.91 (s, 1H), 7.74-7.57 (m, 8H), 7.52 (d, J=6.5, 2H), 4.47 (s, 3H), 4.28-4.06 (m, 6H), 3.98-3.90 (m, 2H), 3.89-3.27 (m, 10H), 3.23-3.05 (m, 4H), 2.94 (br s, 2H).
MS (ESI): mass calcd. for C37H41Cl2N5O4S, 721.2; m/z found, 722.2 [M+H]+. 1H NMR (d6-DMSO): 12.28 (br s, 1H), 10.81 (br s, 2H), 8.73 (s, 1H), 8.74-8.57 (m, 8H), 8.52 (d, J=9.8, 2H), 4.76 (s, 3H), 4.48-4.34 (m, 6H), 4.25-4.00 (m, 3H), 3.91-3.67 (m, 5H), 3.50-2.95 (m, 10H), 1.02 (d, J=7.2, 2H).
MS (ESI): mass calcd. for C37H41Cl2N5O4S, 721.2; m/z found, 722.3 [M+H]+. 1H NMR (d6-DMSO): 9.81 (br s, 1H), 9.69 (br s, 2H), 7.82 (s, 1H), 7.72-7.52 (m, 8H), 7.43 (d, J=6.4, 2H), 4.39 (s, 1H), 4.32 (br s, 2H), 4.20-4.01 (m, 6H), 4.00-3.62 (m, 9H), 3.55-3.25 (m, 5H), 3.18-2.95 (m, 2H), 2.86 (br s, 2H), 1.10 (d, J=7.2, 2H).
MS (ESI): mass calcd. for C36H42Cl2N6O4, 692.3; m/z found, 693.2 [M+H]+. 1H NMR (d6-DMSO): 11.08 (br s, 1H), 9.18 (br s, 2H), 8.06 (s, 1H), 7.92-7.68 (m, 3H), 7.61-7.45 (m, 4H), 4.65 (s, 2H), 4.32-4.17 (m, 2H), 4.16-3.98 (m, 3H), 3.92-3.53 (m, 7H), 3.20-2.90 (m, 5H), 2.89-2.68 (m, 3H), 2.25-2.00 (br s, 2H), 1.98-1.82 (m, 1H), 1.78-1.68 (m, 2H), 1.58-1.45 (m, 1H), 1.23-1.05 (m, 5H).
MS (ESI): mass calcd. for C36H42Cl2N6O4, 692.3; m/z found, 693.2 [M+H]+. 1H NMR (d6-DMSO): 11.08 (br s, 1H), 9.18 (br s, 2H), 8.06 (s, 1H), 7.92-7.68 (m, 3H), 7.61-7.45 (m, 4H), 4.65 (s, 2H), 4.32-4.17 (m, 2H), 4.16-3.98 (m, 3H), 3.92-3.53 (m, 7H), 3.20-2.90 (m, 5H), 2.89-2.68 (m, 3H), 2.25-2.00 (br s, 2H), 1.98-1.82 (m, 1H), 1.78-1.68 (m, 2H), 1.58-1.45 (m, 1H), 1.23-1.05 (m, 5H).
A. 2-Chloro-5-iodo-benzoyl chloride. A stirring solution of 2-chloro-5-iodo-benzoic acid (12.4 g, 43.9 mmol) in SOCl2 (40 mL, 550 mmol) was heated to reflux under a N2 atmosphere. After 20 h, the mixture was concentrated to give the product (13.2 g, 100%) as a yellow solid. 1H NMR (d6-DMSO): 7.88 (s, 1H), 7.65 (dd, J=8.4, 2.0, 1H), 7.14 (d, J=8.4, 1H).
B. 2-Chloro-N-cyclopentyl-5-iodo-benzamide. 2-Chloro-5-iodo-benzoyl chloride (4.1 g, 13.6 mmol) was added to a stirring solution of cyclopentylamine (4.0 mL, 40.5 mmol) in THF (50 mL) at rt. After 1 h, the mixture was concentrated, and the residue was diluted with CH2Cl2 (100 mL) and washed with 0.5 M HCl. The organic layer was dried and concentrated to give the desired product (4.5 g, 94%) as a white solid. MS (ESI): mass calcd. for C12H13ClINO, 349.0; m/z found, 350.0 [M+H]+. 1H NMR (d6-DMSO): 8.22 (d, J=7.2, 1H), 7.52 (d, J=8.4, 1H), 7.4 (s, 1H), 7.04 (d, J=8.4, 1H), 3.98-3.80 (m, 1H), 1.69-1.55 (m, 2H), 1.48-1.38 (m, 2H), 1.37-1.18 (m, 4H).
C. (2-Chloro-5-iodo-benzyl)-cyclopentyl-amine. Borane dimethyl sulfide (3.5 mL, 76.0 mmol) was added via syringe over 5 min to a stirring solution of 2-chloro-N-cyclopentyl-5-iodo-benzamide (4.3 g, 12.3 mmol) in THF (75 mL). Once addition was complete the mixture was heated to 70° C. After 3 h, the mixture was cooled to rt and MeOH (10 mL) was added slowly over 10 min. Once addition of MeOH was complete, 1 M NaOH (10 mL) was added and the mixture was heated to 70° C. After 2 h, the mixture was cooled to rt and concentrated to give a golden oil. The oil was diluted with CH2Cl2 (100 mL) and washed with H2O (75 mL). The organic layer was dried and concentrated to give a clear golden oil. The oil was purified on SiO2 (CH2Cl2 to 5% acetone/CH2Cl2) to provide a colorless oil. The oil was diluted with 1:1 CH2Cl2/Et2O (25 mL) and treated with 1 M HCl/Et2O. The mixture was concentrated to give the desired product (3.3 g, 72%) as a white solid. MS (ESI): mass calcd. for C12H15ClIN, 335.0; m/z found, 336.0 [M+H]+. 1H NMR (d6-DMSO): 9.39 (s, 2H), 8.18 (d, J=2.1, 1H), 7.80 (dd, J=8.4, 2.2, 1H), 7.36 (s, J=8.4, 1H), 4.20 (t, J=6.2, 2H), 3.62-3.46 (m, 2H), 2.10-1.96 (m, 2H), 1.80-1.68 (m, 3H), 1.62-1.46 (m, 2H).
D. The title compound was prepared using methods analogous to those described for Example 249. MS (ESI): mass calcd. for C36H42Cl2N6O3, 676.3; m/z found, 677.3 [M+H]+. 1H NMR (d6-DMSO): 11.15 (br s, 1H), 9.24 (br s, 2H), 8.17 (s, 1H), 8.05-7.80 (m, 3H), 7.77-7.50 (m, 4H), 4.75 (s, 2H), 4.32-4.17 (m, 2H), 4.00-3.68 (m, 6H), 3.67-3.53 (m, 2H), 3.52-3.32 (m, 2H), 3.31-3.05 (m, 2H), 2.98-2.75 (br s, 2H), 2.35-2.15 (m, 2H), 2.12-1.98 (m, 2H), 1.85-1.48 (m, 6H), 1.24 (dd, J=12.4, 6.4, 5H).
MS (ESI): mass calcd. for C34H40Cl2N6O3, 650.3; m/z found, 651.3 [M+H]+. 1H NMR (d6-DMSO): 10.95 (br s, 1H), 9.05 (br s, 2H), 8.07 (s, 1H), 7.92 (s, 1H), 7.85-7.68 (m, 2H), 7.72-7.45 (m, 4H), 4.65 (s, 2H), 4.35-3.98 (m, 4H), 3.95-3.58 (m, 3H), 3.55-3.20 (m, 4H), 3.18-2.68 (m, 7H), 2.25-2.09 (br s, 2H), 1.70-1.50 (m, 2H), 1.19 (dd, J=20.8, 6.4, 4H), 0.86 (t, J=7.6, 3H).
MS (ESI): mass calcd. for C34H40Cl2N6O4, 666.3; m/z found, 667.2 [M+H]+. 1H NMR (d6-DMSO): 11.08 (br s, 1H), 9.08 (br s, 2H), 8.08 (s, 1H), 7.94 (s, 1H), 7.87-7.68 (m, 2H), 7.67-7.43 (m, 4H), 4.78-4.58 (m, 3H), 4.25 (s, 2H), 4.09 (s, 3H), 3.95-3.58 (m, 7H), 3.44 (t, J=5.6, 3H), 3.02 (s, 4H), 2.77 (br s, 2H), 2.30-2.05 (br s, 2H), 1.90-1.69 (m, 2H), 1.12 (br s, 3H).
MS (ESI): mass calcd. for C35H42Cl2N6O5, 696.3; m/z found, 697.4 [M+H]+. 1H NMR (d6-DMSO): 11.08 (br s, 1H), 9.57 (br s, 1H), 8.25-8.05 (m, 2H), 7.94-7.80 (m, 2H), 7.79-7.50 (m, 4H), 5.39 (br s, 2H), 4.73 (s, 2H), 4.65 (s, 2H), 4.25-4.05 (s, 2H), 4.00-3.62 (m, 10H), 3.56-3.46 (m, 2H), 3.22 (s, 3H), 3.07 (br s, 3H), 2.95-2.75 (m, 2H), 2.22 (br s, 2H), 1.24 (d, J=6.8, 4H).
MS (ESI): mass calcd. for C35H41Cl2N7O4, 693.3; m/z found, 694.2 [M+H]+. 1H NMR (d6-DMSO): 10.71 (br s, 1H), 8.51 (t, J=5.6, 1H), 8.38 (br s, 2H), 7.94 (br s, 1H), 7.69 (s, 1H), 7.65-7.55 (m, 2H), 7.54-7.32 (m, 4H), 4.58-4.40 (m, 2H), 4.05-3.88 (m, 2H), 3.78-3.05 (s, 11H), 3.02-2.55 (m, 6H), 2.33 (t, J=5.2, 2H), 2.12-1.92 (m, 2H), 1.70-1.50 (m, 2H), 1.04 (d, J=6.8, 4H).
A. 3-(2-Chloro-5-iodo-benzyloxy)-propan-1-ol. 1,3-Propanediol (700 μL, 9.7 mmol) was added to a stirring mixture of NaH (60% in oil; 0.45 g, 11.3 mmol) and 2-bromomethyl-1-chloro-4-iodo-benzene (2.6 g, 7.7 mmol) in THF (20 mL). The mixture was heated at reflux under a N2 atmosphere for 20 h. The mixture was cooled to rt, diluted with satd. aq. NH4Cl and extracted with CH2Cl2 (2×25 mL). The organic layers were combined, dried, and concentrated to give a golden oil. The oil was purified on SiO2 (CH2Cl2 to 10% acetone/CH2Cl2) to give the desired product (1.5 g, 60%) as a clear light golden oil. TLC: Rf=0.17 [CH2Cl2]. MS (ESI): mass calcd. C10H12ClIO2, 326.0; m/z found, 349.0 [M+Na]+. 1H NMR (d6-DMSO): 7.80 (d, J=2.2, 1H), 7.66 (dd, J=8.4, 2.2, 1H), 7.25 (d, J=8.3, 1H), 4.48 (s, 2H), 4.46 (t, J=5.2, 1H), 3.57 (t, J=6.4, 2H), 3.49 (q, J=6.3, 2H), 1.72 (p, J=6.4, 2H).
B. The title compound was prepared using methods analogous to those described for Example 249. MS (ESI): mass calcd. for C34H39Cl2N5O5, 667.2; m/z found, 668.2 [M+H]+. 1H NMR (d6-DMSO): 8.09 (s, 1H), 7.88-7.68 (m, 2H), 7.65-7.54 (m, 3H), 7.52-7.42 (m, 2H), 4.67 (s, 2H), 4.49 (s, 2H), 4.38 (t, J=5.2, 2H), 3.99 (t, J=6.8, 2H), 3.78-3.68 (m, 2H), 3.67-3.32 (m, 7H), 3.08-2.98 (m, 1H), 2.90-2.70 (m, 2H), 2.68-2.55 (m, 2H), 2.32-2.14 (m, 2H), 1.98-1.75 (m, 2H), 1.67 (p, J=6.4, 2H), 0.77 (d, J=6.0, 3H).
MS (ESI): mass calcd. for C35H40Cl2N6O4, 678.3; m/z found, 679.2 [M+H]+. 1H NMR (d6-DMSO): 10.92 (br s, 1H), 8.91 (br s, 2H), 7.93 (s, 1H), 7.75-7.55 (m, 3H), 7.54-7.44 (m, 2H), 7.40-7.32 (m, 2H), 4.52 (s, 2H), 4.39 (s, 2H), 4.17 (s, 1H), 4.02-3.85 (m, 2H), 3.80-38 (m, 4H), 3.36-2.95 (m, 9H), 2.94-2.75 (m, 2H), 2.70-2.50 (m, 2H), 2.10-1.92 (m, 3H), 1.90-1.66 (m, 1H), 1.00 (d, J=8.2, 3H).
A. 7-Bromo-2-cyclopropyl-1,2,3,4-tetrahydro-isoquinoline. A sample of 7-bromo-1,2,3,4-tetrahydroisoquinoline hydrochloride (1.31 g, 5.27 mmol) was partitioned between 1.0 N NaOH (10 mL) and EtOAc (30 mL). The organic layer was concentrated. A solution of the resulting free amine in MeOH (7 mL), THF (7 mL) and acetic acid (1.5 mL) was treated with (1-ethoxy-cyclopropoxy)-trimethyl-silane (1.84 g, 10.5 mmol) followed by sodium cyanoborohydride (1.09 g. 17.4 mmol). The mixture was heated at 60° C. for 24 h. The mixture was partitioned between EtOAc and satd. aq. NaHCO3. The aqueous layer was extracted with CH2Cl2. The combined organic layers were dried and concentrated. Purification of the residue (SiO2; hexanes to 30% EtOAc/hexanes) afforded the desired product (1.17 g, 88%). 1H NMR (CDCl3): 7.22 (dd, J=8.2, 2.0, 1H), 7.18-7.15 (m, 1H), 6.94 (d, J=8.2, 1H), 3.74 (s, 2H), 2.90 (t, J=5.9, 2H), 2.80 (t, J=5.9, 2H), 1.82-1.74 (m, 4H).
B. The title compound was prepared according to the methods described for Example 134, substituting 7-bromo-2-cyclopropyl-1,2,3,4-tetrahydro-isoquinoline for 7-bromo-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester in Step B. MS (ESI): mass calcd. for C37H45ClN6O2S, 672.3; m/z found, 673.3 [M+H]+. 1H NMR (CDCl3): 7.70 (d, J=1.8, 1H), 7.42-7.36 (m, 2H), 7.29-7.26 (m, 1H), 7.22 (s, 1H), 7.02 (d, J=7.9, 1H), 4.47 (s, 2H), 4.03 (t, J=6.7, 2H), 3.73 (s, 2H), 3.59 (t, J=5.7, 2H), 2.90-2.80 (m, 9H), 2.70-2.26 (m, 10H), 2.11-1.99 (m, 2H), 1.78-1.71 (m, 1H), 1.62-1.55 (m, 1H), 0.50-0.44 (m, 4H), 0.41-0.32 (m, 4H).
The title compound was prepared using methods analogous to those described for Example 276. MS (ESI): mass calcd. for C35H42ClN5O3S, 647.3; m/z found, 648.2 [M+H]+. 1H NMR (CDCl3): 7.77 (d, J=1.9, 1H), 7.50-7.43 (m, 2H), 7.36-7.33 (m, 1H), 7.30-7.29 (m, 1H), 7.09 (d, J=7.9, 1H), 4.54 (s, 2H), 4.15-4.00 (m, 2H), 3.83-3.76 (m, 3H), 3.71-3.59 (m, 4H), 3.28-3.21 (m, 1H), 2.98-2.70 (m, 12H), 2.47-2.36 (m, 1H), 2.32-2.19 (m, 2H), 2.16-1.98 (m, 1H), 1.85-1.78 (m, 1H), 0.92 (d, J=6.30, 3H), 0.57-0.51 (m, 4H).
The title compound was prepared using methods analogous to those described for Example 134. MS (ESI): mass calcd. for C32H38ClN5O3S, 607.2; m/z found, 608.2 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=2.0, 1H), 7.53-7.43 (m, 2H), 7.39-7.35 (m, 1H), 7.32-7.27 (m, 1H), 7.10 (d, J=7.9, 1H), 4.54 (s, 2H), 4.14-4.00 (m, 3H), 3.82-3.75 (m, 1H), 3.70-3.59 (m, 4H), 3.32-3.14 (m, 3H), 2.97-2.70 (m, 10H), 2.46-2.35 (m, 1H), 2.32-2.18 (m, 2H), 2.16-1.97 (m, 2H), 1.77-1.68 (m, 1H), 0.92 (d, J=6.3, 3H).
To a solution of 7-({2-chloro-5-[1-{3-[(3S)-3-methylmorpholin-4-yl]propyl}-5-(methylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl]phenyl}ethynyl)-1,2,3,4-tetrahydroisoquinoline (0.05 g, 0.082 mmol) in CH2Cl2 (0.41 mL) was added acetone (7 mg, 0.121 mmol) followed by sodium triacetoxyborohydride (0.043 g, 0.206 mmol). After 18 h, the mixture was diluted with satd. aq. NaHCO3 and extracted with CH2Cl2. The combined organic layers were dried and concentrated. Purification of the residue (SiO2; CH2Cl2 to 3% MeOH/CH2Cl2) afforded the desired product (0.04 g, 75%). MS (ESI): mass calcd. for C35H44ClN5O3S, 649.3; m/z found, 650.2 [M+H]+. 1H NMR (CDCl3): 7.77 (d, J=1.9, 1H), 7.54-7.41 (m, 2H), 7.36-7.33 (m, 1H), 7.31-7.29 (m, 1H), 7.10 (d, J=7.9, 1H), 4.54 (s, 2H), 4.15-4.00 (m, 2H), 3.84-3.57 (m, 7H), 3.24 (dd, J=11.1, 8.7, 1H), 2.99-2.69 (m, 12H), 2.46-2.35 (m, 1H), 2.31-2.20 (m, 2H), 2.14-1.98 (m, 2H), 1.16 (d, J=6.5, 6H), 0.92 (d, J=6.3, 3H).
The title compound was prepared using methods analogous to those described for Example 276. MS (ESI): mass calcd. for C40H45ClN6O2S, 708.3; m/z found, 709.2 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=1.9, 1H), 7.53-7.43 (m, 2H), 7.37-7.33 (m, 1H), 7.31-7.23 (m, 3H), 7.09 (d, J=7.9, 1H), 6.95-6.91 (m, 2H), 6.89-6.84 (m, 1H), 4.54 (s, 2H), 4.12 (t, J=6.8, 2H), 3.80 (s, 2H), 3.65 (t, J=5.8, 2H), 3.22-3.17 (m, 4H), 2.97-2.87 (m, 9H), 2.60-2.56 (m, 4H), 2.38 (t, J=6.8, 2H), 2.17-2.08 (m, 2H), 1.86-1.77 (m, 1H), 0.58-0.50 (m, 4H).
The title compound was prepared using methods analogous to those described for Example 279. MS (ESI): mass calcd. for C33H40ClN5O3S, 621.3; m/z found, 622.2 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.9, 1H), 7.51-7.42 (m, 2H), 7.38-7.35 (m, 1H), 7.28 (s, 1H), 7.11 (d, J=1.9, 1H), 4.54 (s, 2H), 4.14-4.00 (m, 2H), 3.82-3.76 (m, 1H), 3.72-3.58 (m, 6H), 3.24 (dd, J=11.2, 8.7, 1H), 2.98-2.69 (m, 11H), 2.48 (s, 3H), 2.45-2.36 (m, 1H), 2.31-2.19 (m, 2H), 2.15-1.97 (m, 2H), 0.92 (d, J=6.3, 3H).
The compounds in Examples 282-284 were prepared using methods analogous to those described for Example 276.
MS (ESI): mass calcd. for C35H41ClN6O2, 612.3; m/z found, 613.2 [M+H]+. 1H NMR (CDCl3): 7.77 (d, J=2.1, 1H), 7.52 (dd, J=8.4, 2.1, 1H), 7.45 (d, J=8.4, 1H), 7.36-7.32 (m, 1H), 7.29-7.28 (m, 1H), 7.09 (d, J=8.4, 1H), 4.61-4.54 (m, 4H), 4.14-4.00 (m, 2H), 3.85-3.74 (m, 5H), 3.68-3.59 (m, 2H), 3.24 (dd, J=11.1, 8.8, 1H), 2.97-2.87 (m, 4H), 2.86-2.70 (m, 4H), 2.45-2.36 (m, 1H), 2.31-2.20 (m, 2H), 2.14-1.97 (m, 2H), 1.84-1.78 (m, 1H), 0.92 (d, J=6.3, 3H), 0.57-0.52 (m, 4H).
MS (ESI): mass calcd. for C36H41ClN6O3, 640.3; m/z found, 641.2 [M+H]+. 1H NMR (CDCl3): 7.87-7.79 (m, 1H), 7.55-7.40 (m, 2H), 7.36-7.32 (m, 1H), 7.30-7.28 (m, 1H), 7.09 (d, J=7.9, 1H), 5.67-5.59 (m, 1H), 5.24-5.19 (m, 1H), 4.85-4.77 (m, 1H), 4.33-4.20 (m, 2H), 4.14-3.99 (m, 2H), 3.82-3.75 (m, 3H), 3.70-3.57 (m, 2H), 3.28-3.20 (m, 1H), 3.00-2.69 (m, 9H), 2.46-2.35 (m, 1H), 2.31-2.19 (m, 2H), 2.13-1.98 (m, 2H), 1.84-1.78 (m, 1H), 0.95-0.87 (m, 3H), 0.59-0.49 (m, 4H).
MS (ESI): mass calcd. for C34H40ClN5O3S, 633.3; m/z found, 634.2 [M+H]+. 1H NMR (CDCl3): 7.77 (d, J=1.8, 1H), 7.50-7.43 (m, 2H), 7.37-7.32 (m, 2H), 7.04 (d, J=8.4, 1H), 4.54 (s, 2H), 4.10 (t, J=6.8, 2H), 3.81 (s, 2H), 3.72-3.63 (m, 6H), 2.97-2.86 (m, 9H), 2.45-2.37 (m, 4H), 2.33 (t, J=6.9, 2H), 2.13-2.02 (m, 2H), 1.84-1.77 (m, 1H), 0.57-0.50 (m, 4H).
A. 2-Chloro-5-iodo-benzaldehyde. To a 0° C. solution of 2-chloro-5-iodo-benzonitrile (10.6 g, 40.4 mmol) in toluene (100 mL) was added DiBAL-H (1.5 M in toluene; 40.4 mL, 60.6 mmol). The mixture was stirred at 0° C. for 15 min and then warmed to rt for 3 h. The mixture was poured over 1.0 M H2SO4 (10 mL), MeOH (100 mL), and ice. The mixture was stirred vigorously and then was extracted with EtOAc (200 mL). The organic layer was dried and concentrated to give the title compound (2.98 g, 31%). 1H NMR (CDCl3): 10.37 (s, 1H), 8.22 (d, J=2.2, 1H), 7.83 (dd, J=8.4, 2.2, 1H), 7.21 (d, J=8.4, 1H).
B. Benzyl-(2-chloro-5-iodo-benzyl)-amine. To a slurry of 2-chloro-5-iodo-benzaldehyde (5.1 g, 21.5 mmol) and benzylamine (4.6 g, 43.1 mmol) in dichlorothethane (100 mL) was added sodium triacetoxyborohydride (13.7 g, 64.5 mmol). The reaction mixture was stirred for 18 h, then was diluted with satd. aq. NaHCO3, and extracted with CH2Cl2 (2×). The combined organic extracts were washed with water (3×), dried, and concentrated. Purification by silica gel chromatography (hexanes to 20% EtOAc/hexanes) afforded the desired product (5.48 g, 80%).
C. To a solution of 2-[3-(4-chloro-3-iodo-phenyl)-1-(3-morpholin-4-yl-propyl)-1,4,6,7-tetrahydro-pyrazolo[4,3-c]pyridin-5-yl]-2-oxo-acetamide (prepared as described for Example 158, Step D; 103 mg, 0.180 mmol), PdCl2(PPh)3 (13 mg, 0.018 mmol), trimethylsilylacetylene (21 mg, 0.216 mmol), and CuI (3 mg, 0.018 mmol) in degassed DMF (1 mL) was added Et3N (75 μL, 0.54 mmol). The reaction mixture was stirred under N2 for 18 h. The mixture was then treated with benzyl-(2-chloro-5-iodo-benzyl)-amine (97 mg, 0.270 mmol), DBU (81 μL, 0.54 mmol), and degassed water (25 μL), and the reaction was stirred for an additional 24 h. The mixture was diluted with satd. aq. NaHCO3 and extracted with CH2Cl2 (3×). The combined organic extracts were dried and concentrated. Purification by preparative thin layer chromatography (SiO2; 7% MeOH/NH3/CH2Cl2) afforded the desired product (68 mg, 54%). MS (ESI): mass calcd. for C38H40Cl2N6O3, 698.3; m/z found, 699.2 [M+H]+. 1H NMR (CDCl3): 7.88-7.76 (m, 1H), 7.73-7.64 (m, 1H), 7.59-7.51 (m, 1H), 7.51-7.21 (m, 7H), 7.17-6.99 (m, 1H), 5.78-5.62 (m, 1H), 5.31-5.14 (m, 1H), 4.88-4.75 (m, 1H), 4.32-4.19 (m, 2H), 4.16-3.59 (m, 9H), 3.35-3.22 (m, 1H), 3.06-2.68 (m, 4H), 2.54-1.96 (m, 5H), 0.94 (d, J=5.8, 3H).
The compounds in Examples 286-287 were prepared using methods analogous to those described for Example 285.
MS (ESI): mass calcd. for C37H41Cl2N5O3S, 705.2; m/z found, 706.2 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=1.9, 1H), 7.67 (d, J=1.9, 1H), 7.53-7.41 (m, 3H), 7.40-7.32 (m, 5H), 7.30-7.24 (m, 1H), 4.61-4.46 (m, 2H), 4.17-3.99 (m, 2H), 3.92 (s, 2H), 3.85 (s, 2H), 3.83-3.76 (m, 1H), 3.73-3.59 (m, 4H), 3.25 (dd, J=11.1, 8.8, 1H), 2.98-2.83 (m, 5H), 2.83-2.69 (m, 2H), 2.48-2.36 (m, 1H), 2.33-2.20 (m, 2H), 2.17-1.96 (m, 2H), 0.92 (d, J=6.3, 3H).
MS (ESI): mass calcd. for C36H39Cl2N5O3S, 691.2; m/z found, 692.1 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=1.9, 1H), 7.67 (d, J=1.9, 1H), 7.53-7.41 (m, 3H), 7.41-7.31 (m, 5H), 7.29-7.24 (m, 1H), 4.54 (s, 2H), 4.11 (t, J=6.8, 2H), 3.92 (s, 2H), 3.85 (s, 2H), 3.74-3.63 (m, 6H), 2.93-2.87 (m, 5H), 2.47-2.37 (m, 4H), 2.33 (t, J=6.8, 2H), 2.13-2.03 (m, 2H).
The title compound was prepared using methods analogous to those described for Example 2. MS (ESI): mass calcd. for C36H43Cl2N5O3S, 695.3; m/z found, 696.2 [M+H]+. 1H NMR (CDCl3): 7.45 (d, J=2.0, 1H), 7.41-7.37 (m, 1H), 7.36-7.30 (m, 5H), 7.30-7.23 (m, 3H), 7.08 (dd, J=8.1, 2.2, 1H), 4.44 (s, 2H), 4.09 (t, J=6.8, 2H), 3.87 (s, 2H), 3.78 (s, 2H), 3.74-3.60 (m, 6H), 3.09-3.01 (m, 2H), 2.95-2.84 (m, 7H), 2.44-2.37 (m, 4H), 2.32 (t, J=6.8, 2H), 2.11-2.02 (m, 2H).
The compounds in Examples 289-295 were prepared using methods analogous to those described for Example 285.
MS (ESI): mass calcd. for C37H39Cl2N7O3, 699.3; m/z found, 700.2 [M+H]+. 1H NMR (CDCl3): 8.59-8.55 (m, 1H), 7.86-7.81 (m, 1H), 7.73-7.62 (m, 2H), 7.59-7.52 (m, 1H), 7.51-7.32 (m, 5H), 7.20-7.14 (m, 1H), 7.10-6.99 (m, 1H), 5.87-5.56 (m, 1H), 5.31-5.19 (m, 1H), 4.85-4.80 (m, 1H), 4.35-4.18 (m, 2H), 4.16-3.87 (m, 6H), 3.84-3.74 (m, 1H), 3.72-3.56 (m, 2H), 3.26 (t, J=9.9, 1H), 3.03-2.69 (m, 4H), 2.48-1.99 (m, 5H), 0.92 (d, J=6.2, 3H).
MS (ESI): mass calcd. for C36H39ClFN5O3S, 675.2; m/z found, 676.2 [M+H]+. 1H NMR (CDCl3): 7.78 (d, J=1.9, 1H), 7.64 (dd, J=7.1, 2.1, 1H), 7.53-7.44 (m, 3H), 7.40-7.31 (m, 4H), 7.29-7.25 (m, 1H), 7.05 (dd, J=9.7, 8.5, 1H), 4.54 (s, 2H), 4.11 (t, J=6.8, 2H), 3.88 (s, 2H), 3.84 (s, 2H), 3.73-3.64 (m, 6H), 2.93-2.87 (m, 5H), 2.46-2.39 (m, 4H), 2.34 (t, J=6.8, 2H), 2.14-2.04 (m, 2H).
MS (ESI): mass calcd. for C36H38Cl2N6OS, 672.2; m/z found, 673.2 [M+H]+. 1H NMR (CDCl3): 7.80 (d, J=9.9, 1H), 7.66 (d, J=9.9, 1H), 7.56-7.51 (m, 1H), 7.48-7.31 (m, 7H), 7.30-7.25 (m, 1H), 4.68-4.51 (m, 4H), 4.07 (t, J=6.8, 2H), 3.92 (s, 2H), 3.85 (s, 2H), 3.77 (t, J=5.7, 2H), 2.79 (t, J=5.6, 2H), 2.71-2.62 (m, 8H), 2.35 (t, J=5.6, 2H), 2.10-2.01 (m, 2H).
MS (ESI): mass calcd. for C35H37Cl2N7OS, 673.2; m/z found, 674.2 [M+H]+. 1H NMR (CDCl3): 8.63-8.51 (m, 1H), 7.78 (d, J=2.1, 1H), 7.71-7.63 (m, 2H), 7.57-7.52 (m, 1H), 7.49-7.31 (m, 4H), 7.21-7.15 (m, 1H), 4.66-4.52 (m, 4H), 4.08 (t, J=6.8, 2H), 3.98 (s, 2H), 3.97 (s, 2H), 3.78 (t, J=5.7, 2H), 2.79 (t, J=5.3, 2H), 2.71-2.62 (m, 8H), 2.36 (t, J=6.8, 2H), 2.10-2.00 (m, 2H).
MS (ESI): mass calcd. for C32H39Cl2N5O4S, 659.2; m/z found, 660.2 [M+H]+. 1H NMR (CDCl3): 7.79 (d, J=2.0, 1H), 7.62 (d, J=1.9, 1H), 7.54-7.42 (m, 3H), 7.37 (d, J=8.2, 1H), 4.55 (s, 2H), 4.16-4.00 (m, 2H), 3.93 (s, 2H), 3.82-3.76 (m, 1H), 3.73-3.58 (m, 6H), 3.25 (dd, J=11.2, 8.7, 1H), 2.96-2.70 (m, 9H), 2.47-2.37 (m, 1H), 2.33-2.19 (m, 2H), 2.15-1.98 (m, 2H), 0.92 (d, J=6.3, 3H).
MS (ESI): mass calcd. for C34H38Cl2N6O5, 680.2; m/z found, 681.3 [M+H]+. 1H NMR (CDCl3): 7.87-7.80 (m, 1H), 7.67-7.63 (m, 1H), 7.56 (dd, J=8.4, 2.2, 1H), 7.51-7.40 (m, 2H), 7.39-7.33 (m, 1H), 7.11-7.00 (m, 1H), 5.83-5.58 (m, 1H), 5.31-5.17 (m, 1H), 4.89-4.78 (m, 1H), 4.35-4.21 (m, 2H), 4.15-3.95 (m, 3H), 3.93 (s, 2H), 3.82-3.75 (m, 2H), 3.74 (s, 2H), 3.69-3.56 (m, 2H), 3.48 (s, 2H), 3.28-3.19 (m, 1H), 3.04-2.67 (m, 4H), 2.47-2.33 (m, 1H), 2.31-2.18 (m, 2H), 2.14-1.97 (m, 2H), 0.93-0.89 (m, 3H).
MS (ESI): mass calcd. for C33H38Cl2N6O4, 652.2; m/z found, 653.2 [M+H]+. 1H NMR (CDCl3): 7.83-7.74 (m, 1H), 7.69-7.64 (m, 1H), 7.61-7.39 (m, 4H), 5.17-5.10 (m, 1H), 4.81 (s, 1H), 4.20-3.92 (m, 6H), 3.87-3.59 (m, 5H), 3.41-3.35 (m, 2H), 3.30-3.24 (m, 1H), 3.01-2.72 (m, 6H), 2.52-2.40 (m, 1H), 2.38-2.24 (m, 2H), 2.15-1.96 (m, 2H), 0.96-0.93 (m, 3H).
The compounds in Table 1 (Examples 296-621) were prepared using methods analogous to those described in the preceding examples.
Recombinant human cathepsin S (CatS) was expressed in the baculovirus system and purified in one step with a thiopropyl-sepharose column. 10-L yielded ˜700 mg of CatS and N-terminal sequencing confirmed identity. The assay is run in 150 mM sodium acetate pH 5.0 containing 1.5 mM DTT and 150 mM NaCl. The substrate for the assay is: Z-Valine-Valine-Arginine-AMC (catalog #1-1540, Bachem). The Km for the substrate is around 5 μM but the presence of substrate inhibition makes kinetic analysis difficult. With 10 μM substrate the assay rate is linear over the range of 1-8 ng CatS in 100 μL reaction. Using 2 ng/well of CatS, the production of product is linear and yields-7-fold signal after 20 min with only 20% loss of substrate. Measurements are taken every min for 20 min. The rate is calculated from the slope of the increase in fluorescence and the percent inhibition is calculated from this.
Results for the compounds tested in this assay are presented in Tables 2, 3, and 4 as an average of results obtained. Compounds were tested in free base, hydrochloride salt, trifluoroacetic acid salt, citric acid salt, or formic acid salt forms.
While the invention has been illustrated by reference to examples, it is understood that the invention is intended not to be limited to the foregoing detailed description.
This application claims the benefit of U.S. provisional patent application Ser. No. 60/889,977, filed Feb. 15, 2007 which is incorporated herein by reference.
| Number | Date | Country | |
|---|---|---|---|
| 60889977 | Feb 2007 | US |
