Patent Grant
8420657
This application claims priority to European Serial No. 08151137.0 filed 6 Feb. 2008, the contents of which are incorporated herein by reference in their entirety.
The invention relates to 7-phenyl-7H-pyrrolo[2,3d]pyrimidin-2yl-amino derivatives of the formula I given below, as well as salts thereof; processes for the preparation thereof; pharmaceutical compositions comprising a compound of the formula (I), optionally in the presence of a combination partner; the application of a compound of formula (I) in a process for the treatment of the human or animal body, (in particular with regard to a proliferative disease); the use of a compound of formula (I) for manufacturing a medicament for the treatment of such diseases.
Janus kinases (JAKs) form a family of intracellular protein tyrosine kinases with four members, JAK1, JAK2, JAK3 and TYK2. These kinases are important in the mediation of cytokine receptor signaling which induces various biological responses including cell proliferation, differentiation and apoptosis. Knock-out experiments in mice have shown that JAKs are inter alia important in hematopoiesis. In addition, JAK2 was shown to be implicated in myeloproliferative diseases and cancers. JAK2 activation by chromosome re-arrangements and/or loss of negative JAK/STAT (STAT=signal transducing and activating factor(s)) pathway regulators has been observed in hematological malignancies as well as in certain solid tumors.
WO 2005/080393 discloses inter alia 7H-pyrrolo[2,3d]pyrimidin-2yl-amino derivatives which are useful in the treatment of disorders associated with abnormal or deregulated kinase activity.
Bioorganic & Medical Chemistry Letters 16 (2006), 2689 discloses design and synthesis of certain 7H-pyrrolo[2,3d]pyrimidines as focal adhesion kinase inhibitors.
It has now been found that the 7-phenyl-7H-pyrrolo[2,3d]pyrimidin-2yl-amino derivatives of the formula I given below, have advantageous pharmacological properties and inhibit, for example, the tyrosine kinase activity of Janus kinases, such as JAK2 kinase and/or JAK3- (but also JAK-1-) kinase. Hence, the compounds of formula I are suitable, for example, to be used in the treatment of diseases depending on the tyrosine kinase activity of JAK2 (and/or JAK3) kinase, especially proliferative diseases such as tumor diseases, leukaemias, polycythemia vera, essential thrombocythemia, and myelofibrosis with myeloid metaplasia. Through the inhibition of JAK-3 kinase, compounds of the invention also have utility as immunosuppressive agents, for example for the treatment of diseases such as organ transplant rejection, lupus erythematodes, multiple sclerosis, rheumatoid arthritis, psoriasis, dermatitis, Crohn's disease, type-1 diabetes and complications from type-1 diabetes.
In a first aspect, the invention relates to compounds of the formula I,
wherein
The invention may be more fully appreciated by reference to the following description, including the following glossary of terms and the concluding examples. For the sake of brevity, the disclosures of the publications 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.
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. If at least one asymmetrical carbon atom is present in a compound of the formula I, such a compound may exist in optically active form or in the form of a mixture of optical isomers, e. g. in the form of a racemic mixture. All optical isomers and their mixtures, including the racemic mixtures, are part of the present invention. Thus, any given 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.
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, and chlorine, such as 2H, 3H, 11C, 13C, 14C, 15N, 18F 31P, 32P, 35S, 36Cl, 125I respectively. Various isotopically labeled compounds of the present invention, for example those into which radioactive isotopes such as 3H, 13C, and 14C are incorporated. 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 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 moiety 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 (where one or more up to all more general expressions in embodiments characterized as preferred above or below can be replaced with a more specific definition, thus leading to a more preferred embodiment of the invention, respectively).
Where the plural form (e.g. compounds, salts) is used, this includes the singular (e.g. a single compound, a single salt). “A compound” does not exclude that (e.g. in a pharmaceutical formulation) more than one compound of the formula I (or a salt thereof) is present.
The acid addition salt of compounds of formula I are preferably pharmaceutically acceptable salts. Such salts are known in the field.
The following general definitions shall apply in this specification, unless otherwise specified:
Halogen (or halo) denotes fluorine, bromine, chlorine or iodine, in particular fluorine, chlorine. Halogen-substituted groups and moieties, such as alkyl substituted by halogen (halogenalkyl) can be mono-, poly- or per-halogenated.
Hetero atoms are atoms other than Carbon and Hydrogen, preferably nitrogen (N), oxygen (O) or sulfur (S).
Carbon containing groups, moieties or molecules contain 1 to 8, preferably 1 to 6, more preferably 1 to 4, most preferably 1 or 2, carbon atoms. Any non-cyclic carbon containing group or moiety with more than 1 carbon atom is straight-chain or branched.
The prefix “lower” or “C1-C7” denotes a radical having up to and including a maximum of 7, especially up to and including a maximum of 4 carbon atoms, the radicals in question being either linear or branched with single or multiple branching.
“Alkyl” refers to a straight-chain or branched-chain alkyl group, preferably represents a straight-chain or branched-chain C1-12alkyl, particularly preferably represents a straight-chain or branched-chain C1-7alkyl; for example, methyl, ethyl, n- or iso-propyl, n-, iso-, sec- or tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, with particular preference given to methyl, ethyl, n-propyl, iso-propyl and n-butyl and iso-butyl. Alkyl may be unsubstituted or substituted. Exemplary substituents include, but are not limited to hydroxyl, alkoxy, oxo (i.e. ═O), halogen and amino. An example of a substituted alkyl is trifluoromethyl. Cycloalkyl may also be a substituent to alkyl. An example of such a case is the moiety (alkyl)-cyclopropyl or alkandiyl-cyclopropyl, e.g. —CH2-cyclopropyl. C1-C7-alkyl is preferably alkyl with from and including 1 up to and including 7, preferably from and including 1 up to and including 4, and is linear or branched; preferably, lower alkyl is butyl, such as n-butyl, sec-butyl, isobutyl, tert-butyl, propyl, such as n-propyl or isopropyl, ethyl or preferably methyl.
Each alkyl part of other groups like “alkoxy”, “alkoxyalkyl”, “alkoxycarbonyl”, “alkoxycarbonylalkyl”, “alkylsulfonyl”, “alkylsulfinyl”, “alkylamino”, “halogenalkyl” shall have the same meaning as described in the above-mentioned definition of “alkyl”.
“Alkandiyl” refers to a straight-chain or branched-chain alkandiyl group bound by two different Carbon atoms to the moiety, it preferably represents a straight-chain or branched-chain C1-12 alkandiyl, particularly preferably represents a straight-chain or branched-chain C1-6 alkandiyl; for example, methandiyl (—CH2—), 1,2-ethanediyl (—CH2—CH2—), 1,1-ethanediyl ((—CH(CH3)—), 1,1-, 1,2-, 1,3-propanediyl and 1,1-, 1,2-, 1,3-, 1,4-butanediyl, with particular preference given to methandiyl, 1,1-ethanediyl, 1,2-ethanediyl, 1,3-propanediyl, 1,4-butanediyl.
“Alkendiyl” refers to a straight-chain or branched-chain alkendiyl group bound by two different Carbon atoms to the molecule, it preferably represents a straight-chain or branched-chain C2-6alkandiyl; for example, —CH═CH—, —CH═C(CH3)—, —CH═CH—CH2—, —C(CH3)═CH—CH2—, —CH═C(CH3)—CH2—, —CH═CH—C(CH3)H—, —CH═CH—CH═CH—, —C(CH3)═CH—CH═CH—, —CH═C(CH3)—CH═CH—, with particular preference given to —CH═CH—CH2—, —CH═CH—CH═CH—. Alkendiyl may be substituted or unsubstituted
“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 moieties: cyclopropyl, cyclobutyl, cyclpentyl and cylclohexyl. Cycloalkyl may be unsubstituted or substituted; exemplary substituents are provided in the definition for alkyl.
“Aryl” refers to an aromatic homocyclic ring system with 6 or more carbon atoms; aryl is preferably an aromatic moiety with 6 to 14 ring carbon atoms, more preferably with 6 to 10 ring carbon atoms, such as phenyl or naphthyl, preferably phenyl. Aryl may be unsubstituted or substituted by one or more, preferably up to three, more preferably up to two substituents independently selected from the group consisting of unsubstituted or substituted heterocyclyl as described below, especially pyrrolidinyl, such as pyrrolidino, oxopyrrolidinyl, such as oxo-pyrrolidino, C1-C7-alkyl-pyrrolidinyl, 2,5-di-(C1-C7alkyl)pyrrolidinyl, such as 2,5-di-(C1-C7alkyl)-pyrrolidino, tetrahydrofuranyl, thiophenyl, C1-C7-alkylpyrazolidinyl, pyridinyl, C1-C7-alkylpiperidinyl, piperidino, piperidino substituted by amino or N-mono- or N,N-di-[lower alkyl, phenyl, C1-C7-alkanoyl and/or phenyl-lower alkyl)-amino, unsubstituted or N-lower alkyl substituted piperidinyl bound via a ring carbon atom, piperazino, lower alkylpiperazino, morpholino, thiomorpholino, S-oxo-thiomorpholino or S,S-dioxothiomorpholino; C1-C7-alkyl, amino-C1-C7-alkyl, N—C1-C7-alkanoylamino-C1-C7-alkyl, N—C1-C7-alkanesulfonyl-amino-C1-C7-alkyl, carbamoyl-C1-C7-alkyl, [N-mono- or N,N-di-(C1-C7-alkyl)-carbamoyl]-C1-C7-alkyl, C1-C7-alkanesulfinyl-C1-C7-alkyl, C1-C7-alkanesulfonyl-C1-C7-alkyl, phenyl, naphthyl, mono- to tri-[C1-C7-alkyl, halo and/or cyano]-phenyl or mono- to tri-[C1-C7-alkyl, halo and/or cyano]-naphthyl; C3-C8-cycloalkyl, mono- to tri-[C1-C7-alkyl and/or hydroxy]-C3-C8-cycloalkyl; halo, hydroxy, lower alkoxy, lower-alkoxy-lower alkoxy, (lower-alkoxy)-lower alkoxy-lower alkoxy, halo-C1-C7-alkoxy, phenoxy, naphthyloxy, phenyl- or naphthyl-lower alkoxy; amino-C1-C7-alkoxy, lower-alkanoyloxy, benzoyloxy, naphthoyloxy, formyl (CHO), amino, N-mono- or N,N-di-(C1-C7-alkyl)-amino, C1-C7-alkanoylamino, C1-C7-alkanesulfonylamino, carboxy, lower alkoxy carbonyl, e.g.; phenyl- or naphthyl-lower alkoxycarbonyl, such as benzyloxycarbonyl; C1-C7-alkanoyl, such as acetyl, benzoyl, naphthoyl, carbamoyl, N-mono- or N,N-disubstituted carbamoyl, such as N-mono- or N,N-di-substituted carbamoyl wherein the substituents are selected from lower alkyl, (lower-alkoxy)-lower alkyl and hydroxy-lower alkyl; amidino, guanidino, ureido, mercapto, lower alkylthio, phenyl- or naphthylthio, phenyl- or naphthyl-lower alkylthio, lower alkyl-phenylthio, lower alkyl-naphthylthio, halogen-lower alkylmercapto, sulfo (—SO3H), lower alkanesulfonyl, phenyl- or naphthyl-sulfonyl, phenyl- or naphthyl-lower alkylsulfonyl, alkylphenylsulfonyl, halogen-lower alkylsulfonyl, such as trifluorome-thanesulfonyl; sulfonamido, benzosulfonamido, azido, azido-C1-C7-alkyl, especially azido-methyl, C1-C7-alkanesulfonyl, sulfamoyl, N-mono- or N,N-di-(C1-C7-alkyl)-sulfamoyl, morpholinosulfonyl, thiomorpholinosulfonyl, cyano and nitro; where each phenyl or naphthyl (also in phenoxy or naphthoxy) mentioned above as substituent or part of a substituent of substituted alkyl (or also of substituted aryl, heterocyclyl etc. mentioned herein) is itself unsubstituted or substituted by one or more, e.g. up to three, preferably 1 or 2, substituents independently selected from halo, halo-lower alkyl, such as trifluoromethyl, hydroxy, lower alkoxy, azido, amino, N-mono- or N,N-di-(lower alkyl and/or C1-C7-alkanoyl)-amino, nitro, carboxy, lower-alkoxycarbonyl, carbamoyl, cyano and/or sulfamoyl.
“Heterocyclyl” refers to a heterocyclic radical that is unsaturated (=carrying the highest possible number of conjugated double bonds in the ring(s) i.e. heteroaryl), saturated or partially saturated and is preferably a monocyclic or in a broader aspect of the invention bicyclic, tricyclic or spirocyclic ring; and has 3 to 24, more preferably 4 to 16, most preferably 5 to 10 and most preferably 5 or 6 ring atoms; wherein one or more, preferably one to four, especially one or two carbon ring atoms are replaced by a heteroatom, the bonding ring preferably having 4 to 12, especially 5 to 7 ring atoms. The heterocyclic radical (heterocyclyl) may be unsubstituted or substituted by one or more, especially 1 to 3, substituents independently selected from the group consisting of the substituents defined above for substituted alkyl and/or from one or more of the following substituents: oxo (═O), thiocarbonyl (═S), imino(═NH), imino-lower alkyl. Further, heterocyclyl is especially a heterocyclyl radical selected from the group consisting of oxiranyl, azirinyl, aziridinyl, 1,2-oxathiolanyl, thienyl (=thiophenyl), furanyl, tetrahydrofuryl, pyranyl, thiopyranyl, thianthrenyl, isobenzofuranyl, benzofuranyl, chromenyl, 2H-pyrrolyl, pyrrolyl, pyrrolinyl, pyrrolidinyl, imidazolyl, imidazolidinyl, benzimidazolyl, pyrazolyl, pyrazinyl, pyrazolidinyl, thiazolyl, isothiazolyl, dithiazolyl, oxazolyl, isoxazolyl, pyridyl, pyrazinyl, pyrimidinyl, piperidinyl, piperazinyl, pyridazinyl, morpholinyl, thiomorpholinyl, (S-oxo or S,S-dioxo)-thiomorpholinyl, indolizinyl, azepanyl, diazepanyl, especially 1,4-diazepanyl, isoindolyl, 3H-indolyl, indolyl, benzimidazolyl, cumaryl, indazolyl, triazolyl, tetrazolyl, purinyl, 4H-quinolizinyl, isoquinolyl, quinolyl, tetrahydroquinolyl, tetrahydroisoquinolyl, decahydroquinolyl, octahydroisoquinolyl, benzofuranyl, dibenzofuranyl, benzothiophenyl, dibenzothiophenyl, phthalazinyl, naphthyridinyl, quinoxalyl, quinazolinyl, quinazolinyl, cinnolinyl, pteridinyl, carbazolyl, beta-carbolinyl, phenanthridinyl, acridinyl, perimidinyl, phenanthrolinyl, furazanyl, phenazinyl, phenothiazinyl, phenoxazinyl, chromenyl, isochromanyl, chromanyl, benzo[1,3]dioxol-5-yl and 2,3-dihydro-benzo[1,4]dioxin-6-yl, each of these radicals being unsubstituted or substituted by one or more, preferably up to three, substituents selected from those mentioned above for substituted aryl and/or from one or more of the following substituents: oxo (═O), thiocarbonyl (═S), imino(═NH), imino-lower alkyl.
“Arylalkyl” refers to an aryl group bound to the molecule via an alkyl group, such as a methyl or ethyl group, preferably phenethyl or benzyl, in particular benzyl. Similarly, cycloalkylalkyl and heterocyclylalkyl represents a cycloalkyl group bound to the molecule via an alkyl group or a heterocyclyl group bound to the molecule via an alkyl group. In each instance, aryl, heterocyclyl, cycloalkyl and alkyl may be substituted as defined above.
“Treatment” includes prophylactic (preventive) and therapeutic treatment as well as the delay of progression of a disease or disorter.
“Protein tyrosine kinase mediated diseases” (especially JAK2 and/or JAK3 kinase mediated diseases) are especially such disorders that respond in a beneficial way (e.g. amelioration of one or more symptoms, delay of the onset of a disease, up to temporary or complete cure from a disease) to the inhibition of a protein tyrosine kinase, especially inhibition of a JAK (preferably JAK2 and/or JAK3) kinase or TYK2, more especially inhibition of JAK2 kinase (where among the diseases to be treated, especially proliferative diseases such as tumor diseases, leukaemias, polycythemia vera, essential thrombocythemia, and myelofibrosis with myeloid metaplasia may be mentioned) and/or of JAK3 kinase (where preferably the treatment (e.g. by immunosuppression) of diseases such as organ transplant rejection, lupus erythematodes, multiple sclerosis, rheumatoid arthritis, psoriasis, dermatitis, Crohn's disease, type-1 diabetes and complications from type-1 diabetes are to be mentioned as preferred.
“Salts” (which, what is meant by “or salts thereof” or “or a salt thereof”, can be present alone or in mixture with free compound of the formula I) are preferably pharmaceutically acceptable salts. Such salts are formed, for example, as acid addition salts, preferably with organic or inorganic acids, from compounds of formula I with a basic nitrogen atom, especially the pharmaceutically acceptable salts. Suitable inorganic acids are, for example, halogen acids, such as hydrochloric acid, sulfuric acid, or phosphoric acid. Suitable organic acids are, e.g., carboxylic acids or sulfonic acids, such as fumaric acid or methansulfonic acid. For isolation or purification purposes it is also possible to use pharmaceutically unacceptable salts, for example picrates or perchlorates. For therapeutic use, only pharmaceutically acceptable salts or free compounds are employed (where applicable in the form of pharmaceutical preparations), and these are therefore preferred. In view of the close relationship between the novel compounds in free form and those in the form of their salts, including those salts that can be used as intermediates, for example in the purification or identification of the novel compounds, any reference to the free compounds hereinbefore and hereinafter is to be understood as referring also to the corresponding salts, as appropriate and expedient.
Combination refers to either a fixed combination in one dosage unit form, or a kit of parts for the combined administration where a compound of the formula I and a combination partner (e.g. an other drug as explained below, also referred to as “therapeutic agent” or “co-agent”) may be administered independently at the same time or separately within time intervals, especially where these time intervals allow that the combination partners show a cooperative, e.g. synergistic effect. The terms “co-administration” or “combined administration” or the like as utilized herein are meant to encompass administration of the selected combination partner to a single subject in need thereof (e.g. a patient), and are intended to include treatment regimens in which the agents are not necessarily administered by the same route of administration or at the same time. The term “pharmaceutical combination” as used herein means a product that results from the mixing or combining of more than one active ingredient and includes both fixed and non-fixed combinations of the active ingredients. The term “fixed combination” means that the active ingredients, e.g. a compound of formula I and a combination partner, are both administered to a patient simultaneously in the form of a single entity or dosage. The term “non-fixed combination” means that the active ingredients, e.g. a compound of formula I and a combination partner, are both administered to a patient as separate entities either simultaneously, concurrently or sequentially with no specific time limits, wherein such administration provides therapeutically effective levels of the two compounds in the body of the patient. The latter also applies to cocktail therapy, e.g. the administration of three or more active ingredients.
In preferred embodiments, which are preferred independently, collectively or in any combination or sub-combination, the invention relates to a compound of the formula I, in free base form or in acid addition salt form, wherein the substituents are as defined herein.
In an embodiment, the invention relates to a compound of formula IA
wherein the substituents are as defined for a compound of formula I.
In a further embodiment, the invention relates to a compound of formula IB
wherein the substituents are as defined for a compound of formula I.
In a further embodiment, the invention relates to a compound of formula IC
wherein the substituents are as defined for a compound of formula I.
In a further embodiment, the invention relates to a compound of formula ID
wherein the substituents are as defined for a compound of formula I.
In a further embodiment, the invention relates to a compound of formula IE
wherein the substituents are as defined for a compound of formula I.
In a further embodiment, the invention relates to compounds of the formula (I) wherein R2a and R3a are both H, that is, this embodiment relates to compounds of the formula I′:
wherein
In a further embodiment, the invention relates to a compound of formula I′A
wherein the substituents are as defined for a compound of formula I.
In a further embodiment, the invention relates to a compound of formula I′B
wherein the substituents are as defined for a compound of formula I.
In a further embodiment, the invention relates to a compound of formula I′C
wherein the substituents are as defined for a compound of formula I.
In a further embodiment, the invention relates to a compound of formula I′D
wherein the substituents are as defined for a compound of formula I.
In a further embodiment, the invention relates to a compound of formula I′E
wherein the substituents are as defined for a compound of formula I.
The following definitions apply to any of the formulae described herein.
In any of the formulae herein, R1 is preferably aryl or heteroaryl, each being unsubstituted or substituted as defined herein.
In a further advantageous embodiment, n represents 0 and R8 represents —SO2CH3.
In a yet further advantageous embodiment, n represents 0 or 1 and R4 represents the group:
as defined above, more preferably wherein the group is as defined above and R6 and R7 together with the nitrogen atom to which they are attached form a morpholinyl, piperazinyl, thiazidinyl or pyrrolidinyl group, each being optionally substituted, most preferably R6 and R7 together with the nitrogen atom to which they are attached form an optionally substituted morpholinyl group, wherein said substituents are selected from the group consisting of oxo (e.g. in C═O or SO2), hydroxy, halo, C1-C7-alkyl, C1-C7-alkyloxy, C1-C7-haloalkyl, C1-C7-haloalkyloxy, C3-C6-cycloalkyl, C3-C6-cycloalkyloxy, C3-C6-halocycloalkyl, C3-C6-halocycloalkyloxy hydroxy-C1-C7-alkyl, C1-C7-alkylamino, di-C1-C7-alkylamino
In a further advantageous embodiment, at least one of R2 and R3 does not represent hydrogen.
In a further advantageous embodiment, R1 is optionally substituted aryl selected from the group consisting of phenyl, naphthyl, each of which is unsubstituted or substituted by one to three moieties independently selected from the group consisting of C1-C7-alkyl, amino-C1-C7-alkyl, halo-C1-C7-alkyl, N—C1-C7-alkanoylamino-C1-C7-alkyl, N—C1-C7-alkanesulfonyl-amino-C1-C7-alkyl, pyrrolidino-C1-C7-alkyl, oxo-pyrrolidino-C1-C7-alkyl, piperidino-C1-C7-alkyl, piperazin-1-yl-C1-C7-alkyl, 4-(C1-C7-alkyl, C1-C7-alkoxy-C1-C7-alkyl, halo-C1-C7-alkyl or C3-C10-cycloalkyl)-piperazin-1-yl-C1-C7-alkyl, 4-(amino-C1-C7-alkyl)-piperazin-1-yl-C1-C7-alkyl, 4-[N-mono- or N,N-di-(C1-C7-alkylamino)-C1-C7-alkyl]-piperazin-1-yl-C1-7-alkyl, morpholino-C1-C7-alkyl, thiomorpholino-C1-C7-alkyl, S-mono- or S,S-dioxo-thiomorpholino-C1-C7-alkyl, carbamoyl-C1-C7-alkyl, [N-mono- or N,N-di-(C1-C7-alkyl)-carbamoyl]-C1-C7-alkyl, C1-C7-alkanesulfinyl-C1-C7-alkyl, C1-C7-alkanesulfonyl-C1-C7-alkyl, halo, hydroxyl, C1-C7-alkoxy, amino, N-mono- or N,N-di-(C1-C7-alkyl)-amino, C1-C7-alkanoylamino, pyrrolidino, oxo-pyrrolidino, piperidino, piperazin-1-yl, 4-(C1-C7-alkyl, C1-C7-alkoxy-C1-C7-alkyl, halo-C1-C7-alkyl or C3-C10-cycloalkyl)-piperazin-1-yl, 4-(amino-C1-C7-alkyl)-piperazin-1-yl, 4-[N-mono- or N,N-di-(C1-C7-alkylamino)-C1-C7-alkyl]-piperazin-1-yl, morpholino, thiomorpholino, S-oxo- or S,S-dioxothiomorpholino, C1-C7-alkanesulfonylamino, carbamoyl, N-mono- or N,N-di-(C1-C7-alkyl, C1-C7-alkoxy-C1-C7-alkyl, amino-C1-C7-alkyl and/or (N′-mono- or N′,N′-di-(C1-C7-alkyl)-amino-C1-C7-alkyl)-carbamoyl, pyrrolidin-1-carbonyl, piperidin-1-carbonyl, piperazin-1-carbonyl, 4-(C1-C7-alkyl)piperazin-1-carbonyl, morpholin-1-carbonyl, thiomorpholin-1-carbonyl, S-oxo- or S,S-dioxothiomorpholin-1-carbonyl, sulfo, C1-C7-alkanesulfonyl, C1-C7-alkanesulfinyl, sulfamoyl, N-mono- or N,N-di-(C1-C7-alkyl)-sulfamoyl, morpholinosulfonyl, thiomorpholinosulfonyl, cyano and nitro; for example, it can preferably be phenyl or naphthyl that is substituted by one or more, especially one to four substituents independently selected from the group consisting of C1-C7-alkoxy, carbamoyl, N-mono- or N,N-di-(C1-C7-alkyl and/or C1-C7-alkyloxy-C1-C7-alkyl)-carbamoyl, N-mono- or N,N-di-{[unsubstituted, N′-mono- or N′,N′-di-(C1-C7-alkyl)-substituted]-carbamoyl, sulfamoyl, N-mono- or N,N-di-(C1-C7-alkyl)-sulfamoyl, piperidino, piperazino, N—C1-C7-alkylpiperazino, morpholino, thiomorpholino, S-oxothiomorpholino and S,S-dioxothiomorpholino, in the case of R2, unsubstituted or substituted aryl is preferably phenyl or naphthyl that is unsubstituted or substituted by one or more, especially up to three, more especially up to two, substituents, preferably not in ortho-position, more preferably with not more than one substituent in meta-position, most preferably with one substituent in meta- and/or one substituent in para position, most preferably with one substituent in meta-position or especially one in para-position, where the substituents are independently selected from the group consisting of C1-C7-alkyl, amino-C1-C7-alkyl, N—C1-C7-alkanoylamino-C1-C7-alkyl, N—C1-C7-alkanesulfonyl-amino-C1-C7-alkyl, carbamoyl-C1-C7-alkyl, [N-mono- or N,N-di-(C1-C7-alkyl)-carbamoyl]-C1-C7-alkyl, C1-C7-alkanesulfinyl-C1-C7-alkyl, C1-C7-alkanesulfonyl-C1-C7-alkyl, halo, hydroxyl, C1-C7-alkoxy, amino, N-mono- or N,N-di-(C1-C7-alkyl)-amino, C1-C7-alkanoylamino, C1-C7-alkanesulfonylamino, carbamoyl, N-mono- or N,N-di-(C1-C7-alkyl)-carbamoyl, C1-C7-alkanesulfonyl, sulf-amoyl, N-mono- or N,N-di-(C1-C7-alkyl)-sulfamoyl, morpholinosulfonyl, thiomorpholino-sulfonyl, cyano and nitro.
In a further advantageous embodiment, R1 is optionally substituted aryl selected from the group consisting of phenyl, (especially 3,4,5-)trimethoxyphenyl*, (especially 3,4- or 3,5-)dimethoxyphenyl*, (especially 4-)morpholinophenyl, (especially 4-)N-(2-methoxyethyl)-carbamoylphenyl*, or (especially 4-)N,N-(2-dimethylamino-ethyl)-carbamoylphenyl*, (especially 4-)dimethylaminocarbonyl-(especially 3-)methyl-phenyl*, (especially 4-)-(preferably 4-)-(2-methoxy-ethyl-piperazin-(especially 1-)yl-(especially 3-)-methyl-phenyl, (especially 4-)pyrrolidin-1-carbonyl-(especially 3-)methyl-phenyl*, (especially 3-)methyl-(especially 4-)-4-methylpiperazin-1-carbonyl-phenyl, (especially 3- or 4-)4-methyl-piperazin-1-yl-phenyl*, (especially 4-)-4-ethyl-piperazin-1-yl-(especially 3-) methyl-phenyl*, (especially 4-)-4-methylpiperazin-1-yl-(especially 3-)cyano-phenyl, (especially 4-)-piperazin-1-yl-phenyl, (especially 4-)-4-cyclopropyl-piperazin-1-yl-phenyl, (especially 4-)-4-(2-dimethylaminoethyl)-piperazin-1-yl-(especially 3-)methyl-phenyl*, (especially 4-)4-isopropyl-piperazin-1-yl)-(especially 3-)methyl-phenyl*, (especially 4-)N,N-diethylaminocarbonyl-(especially 3-)methyl-phenyl*, (especially 4-)4-ethylpiperazin-1-carbonyl-(especially 3-)methyl-phenyl, (especially 4-)-(4-ethylpiperazin-1-ylmethyl)-(especially 3-)methyl-phenyl, (especially 4-)N-methylaminocarbonyl-(especially 3-)methylphenyl, (especially 4-)-4-(3,3,3-trifluoropropyl)-piperazin-1-yl-(especially 3-)methyl-phenyl, (especially 4-)-4-(2-(N′,N′-dimethylamino)ethyl-aminocarbonyl-(especially 3-)methyl-phenyl, (especially 4-)-methanesufonyl-phenyl*, (especially 4-)[(especially 2-)-oxo-pyrrolidin-1-yl]-phenyl, (especially 4-)N,N-diethylaminocarbonyl-(especially 3-)methoxyphenyl, (especially 3-)-4-methylpiperazin-1-yl-(especially 4-)methyl-phenyl, (especially 3-)-4-methylpiperazin-1-yl-(especially 4-)methoxy-phenyl*, (especially 3- or 4-)-morpholinomethyl-(especially 4- or 3-)methyl-phenyl, (especially 2-)acetylamino-indan-(especially 5-)yl, (especially 2-)oxo-2,3-dihydroindol-(especially 5-)yl, (especially 4-)methylsulfinylphenyl, (especially 4-)methoxyphenyl, (especially 4-)methyl-(especially 3-)methoxyphenyl, (especially 4-)-N-(2-methoxyethyl)-aminocarbonyl-phenyl, (especially 4-)N,N-dimethylcarbamoyl-phenyl, (especially 3-)methanesulfonylamino-phenyl, (especially 4-)methoxycarbonyl-(especially 3-)methoxy-phenyl, (especially 4-)N,N-dimethylcarbamoyl-(especially 3-)methoxy-phenyl, (especially 4-)-(4-cyclopropyl-piperazin-1-yl)-(especially 3-)methyl-phenyl*, (especially 4-)-N-(2-(N′,N′-dimethylaminoethyl)-N-methyl-carbamoyl-(especially 3-)methyl-phenyl*, 1,3-dimethyl-oxo-1 H-pyridine-5-yl, (especially 3- or 4-)morpholino-(especially 4- or 3-)methyl-phenyl*, (especially 4-)morpholinomethyl-(especially 3-)methyl-phenyl, (especially 4-)morpholin-1-carbonyl-(especially 3-)methyl-phenyl, (especially 4-)-N-2-(methoxyethyl)aminocarbonyl-(especially 3-)methyl-phenyl, (especially 4-)-N-(3-N′.N′-dimethylaminopropyl)amino-carbonyl-(especially 3-)methyl-phenyl, (especially 5-)-methyl-(especially 6-)methoxy-pyridin-3-yl, (especially 4-)dimethylcarbamoyl-(especially 3,5-)dimethyl-phenyl, (especially 4-)dimethylcarbamoyl-(especially 3-)ethyl-phenyl, (especially 4-(4-)N,N-dimethylcarbamoyl-(especially 3-)methyl-phenyl or (especially 4-)morpholino-(especially 3-)cyano-phenyl; where the moieties marked with an asterisk (*) are especially preferred, as are the moieties where the position after “especially” is given).
In a further advantageous embodiment, R1 is optionally substituted heterocyclyl selected from the group consisting of preferably pyridyl, pyrimidyl, pyrazolyl, thiophenyl, pyrrolyl imidazolyl, or 1H-benzoimidazolyl, each of which is unsubstituted or substituted by one to three moieties independently selected from those mentioned above as substituents for aryl R1, or especially from the group consisting of C1-C7-alkyl, amino-C1-C7-alkyl, N—C1-C7-alkanoylamino-C1-C7-alkyl, N—C1-C7-alkanesulfonyl-amino-C1-C7-alkyl, carbamoyl-C1-C7-alkyl, [N-mono- or N,N-di-(C1-C7-alkyl)-carbamoyl]-C1-C7-alkyl, C1-C7-alkanesulfinyl-C1-C7-alkyl, C1-C7-alkanesulfonyl-C1-C7-alkyl, halo, hydroxyl, C1-C7-alkoxy, amino, N-mono- or N,N-di-(C1-C7-alkyl)-amino, C1-C7-alkanoylamino, C1-C7-alkanesulfonylamino, carbamoyl, N-mono- or N,N-di-(C1-C7-alkyl)-carbamoyl, C1-C7-alkanesulfonyl, sulfamoyl, N-mono- or N,N-di-(C1-C7-alkyl)-sulfamoyl, morpholinosulfonyl, thiomorpholinosulfonyl, thiomorpholinosulfinyl, cyano and nitro.
In a further advantageous embodiment, R1 represents phenyl or pyridiyl in each case optionally substituted by one or two substituents, the substituents being selected from the group consisting of C1-C4-alkyl (in particular methyl), C1-C4-alkoxy (in particular methoxy), halo (in particular fluoro), N-methyl-N-piperazinyl-methyl, N-methyl-N-piperazinyl-carbonyl, 3,5-dimethyl-N-piperazinyl.
The invention further relates to pharmaceutically acceptable prodrugs of a compound of formula (I).
The invention further relates to pharmaceutically acceptable metabolites of a compound of formula (I).
The invention relates especially to the compounds of the formula I given in the Examples, as well as the methods of manufacture described therein.
The compounds of formula I thereof have valuable pharmacological properties, as described hereinbefore and hereinafter. They inhibit various protein tyrosine kinases, and especially JAK2 and/or JAK3-receptor tyrosine kinase. Preferably, the compounds of formula I exhibit selectivity for JAK2 and JAK3 over JAK1 and TYK2 kinases. Most preferably, the compounds of formula I exhibit selectivity for JAK2 over the other JAK family kinases, i.e. preferably, selectivity is exhibited for JAK2 over JAK1, JAK3 and TYK2 kinases.
Preferably, the compounds of formula I exhibit selectively for JAK2 inhibition when compared to other kinases, for example cMet, cKit, ALK and/or PDGFRa.
The compounds of formula I typically show IC50 values for JAK2 inhibition in the range <0.003 to 2 umol l−1, preferably <0.003 to 1 umol l−1, more preferably <0.003 to 0.100 umol l−1, even more preferably <0.003 to 0.050 umol l−1.
The efficacy of the compounds of the invention as inhibitors of JAK/TYK kinase activity can be demonstrated as follows (Results are given at the end of the specification):
All four kinases of the JAK/TYK-kinase family were used as purified recombinant GST-fusion proteins, containing the active kinase domains. GST-JAK1 (866-1154), GST-JAK3(811-1124), and GST-TYK2(888-1187) were expressed and purified by affinity chromatography at the EPK biology unit. GST-JAK2(808-1132) was purchased from Invitrogen (Carlsbad, USA, #4288).
The kinase assays were based on the Caliper mobility shift assay using the LabChip 3000 systems. This technology is similar to capillary electrophoresis and uses charge driven separation of substrate and product in a microfluidic chip.
All kinase reactions were performed in 384 well microtiter plates in a total reaction volume of 18 μl. The assay plates were prepared with 0.1 μl per well of test compound in the appropriate test concentration, as described under the section “preparation of compound dilutions”. The reactions were started by combining 9 μl of substrate mix (consisting of peptide and ATP) with 9 μl of kinase dilution. The reactions were incubated for 60 minutes at 30° C. and stopped by adding 70 μl of stop buffer (100 mM Hepes, 5% DMSO, 0.1% Coating reagent, 10 mM EDTA, 0.015% Brij 35).
Fluorescently labeled synthetic peptides were used as substrates in all reactions. A peptide derived from the sequence of IRS-1(IRS-1 peptide, SEQ ID NO: 1 FITC-Ahx-KKSRGDYMTMQIG-NH2) was used for JAK1 and TYK2 and a peptide named JAK3tide SEQ ID NO: 2 (FITC-GGEEEEYFELVKKKK-NH2) for JAK2 and JAK3. Specific assay conditions are described in Table1:
The terminated reactions were transferred to the Caliper LabChip 3000 reader and the turnover of each reaction was measured by determining the substrate/product ratio.
Preparation of Compound Dilutions
Test compounds were dissolved in DMSO (10 mM) and transferred into 1.4 mL flat bottom or V-shaped Matrix tubes carrying a unique 2D matrix chip by individual compound hubs. The numbers of these chips were distinctively linked to the individual compound identification numbers. The stock solutions were stored at −20° C. if not used immediately. For the test procedure the vials were defrosted and identified by a scanner whereby a working sheet was generated that guided the subsequent working steps. Compound dilutions were made in 96 well plates. This format enabled the assay of maximally 40 individual test compounds at 8 concentrations (single points) including 4 reference compounds. The dilution protocol included the production of pre-dilution plates, master plates and assay plates:
Pre-dilution plates: 96 polypropylene well plates were used as pre-dilution plates. A total of 4 pre-dilution plates were prepared including 10 test compounds each on the plate positions A1-A10, one standard compound at A11 and one DMSO control at A12. All dilution steps were done on a HamiltonSTAR robot.
Master plates: 100 μL of individual compound dilutions including standard compound and controls of the 4 “pre-dilution plates” were transferred into a 384 “master plate” including the following concentrations 1,820, 564, 182, 54.6, 18.2, 5.46, 1.82 and 0.546 μM, respectively in 90% of DMSO.
Assay plates: Identical assay plates were then prepared by pipetting 100 nL each of compound dilutions of the master plates into 384-well “assay plates”. In the following the compounds were mixed with 9 μL of assays components plus 9 μL enzyme corresponding to a 1:181 dilution steps enabling the final concentration of 10, 3.0, 1.0, 0.3, 0.1, 0.03, 0.01 and 0.003 μM, respectively. The preparation of the master plates were handled by the Matrix PlateMate Plus robot and replication of assay plates by the HummingBird robot.
On the basis of these studies, a compound of the invention shows therapeutic efficacy especially against disorders dependent on protein kinase, especially proliferative diseases mediated by JAK/TYK kinase activity.
STAT Nuclear Translocation Assays:
Alternatively, the activity of the compounds of the invention as inhibitors of the JAK/STAT pathway can be demonstrated as follows (Results are given at the end of the specification):
The medium-throughput (96-well format) cellular automated fluorescence microscopy Cellomics assay can be routinely used to assess the functional activation of Janus Kinases (JAKs), based on the nuclear translocation of their substrate, Signal Transducer and Activator of Transcription (STAT). Nuclear translocation can be monitored either in HT1080 fibrosarcoma cells stably transfected with STAT1 fused to Green Fluorescence Protein (GFP) or in U-2 OS osteosarcoma cells stably transfected with STAT5 fused to Green Fluorescence Protein (GFP). Stimulation of HT1080 cells with interferon-γ (IFN-γ) results in JAK1/JAK2-dependent nuclear translocation of STAT1-GFP, whereas stimulation of U-2 OS cells with recombinant human erythropoietin (rhEpo) results in JAK2-dependent nuclear translocation of STAT5-GFP, both of which can be quantified using the Cellomics Cyto/NucTrans software package. This assay may be used to provide an assessment of the nuclear-cytoplasmic differential (NCD) of STAT-GFP using Hoechst dye to define the boundaries of the nucleus.
Generation of HT1080 Fibrosarcoma Cells Stably Expressing GFP-STAT1:
HT1080 fibrosarcoma cells may be obtained from ATCC and can be cultured in alpha Modified Eagle Medium with 10% FCS. Cells can be transfected with pEGFP-N2 STAT1 using Fugene 6 Transfection Reagent following the manufacturers' protocol. 24 hours after transfection the medium can be replaced and selected in 1 mg/ml Geneticin.
Generation of U2OS Cells Stably Expressing STAT5a-GFP and EpoR:
U-2 OS osteosarcoma cells may be obtained from ATCC and can be cultured in standard RPMI medium supplemented with 10% FCS and 2 mM L-glutamine. Cells can be stably transfected with STAT5a-GFP using Lipofectamine following the manufacturers' protocol. 24 hours after transfection the medium can be replaced and selected in 400 μg/ml Geneticin. Subsequently, cells can be stably transfected with EpoR using Lipofectamine following the manufacturers' protocol. 24 hours after transfection the medium can be replaced and cells selected in 100 μg/ml Hygromycin B.
Preparation of Compound Stocks:
Compounds can be dissolved in DMSO to a final stock concentration of 10 mM and stored as aliquots at 4° C. Compounds may be pre-diluted in 100% DMSO at 10 mM, 3 mM 1 mM, 0.3 mM, 0.1 mM, 0.03 mM, 0.01 mM and 0.003 mM. Subsequently, compounds may be diluted in medium and added in 50 μl to the cells. The final compound concentrations tested may be 10 μM, 3 μM 1 μM, 0.3 μM, 0.1 μM, 0.03 μM, 0.01 μM and 0.003 μM and the final DMSO concentration can be 0.1%.
Cellomics Nuclear STAT-GFP Translocation Assays:
HT1080 STAT1-GFP cells may be seeded at a density of 10,000 cells per well in clear-bottom black 96-well Packard View-Plates™. 16-24 hours later, the cells can be treated for 2 hours with 100 ng/ml IFN-γ, washed twice in pre-warmed PBS and fixed in 200 μl of pre-warmed fixation solution (PBS, 3.7% Formaldehyde) for 10 minutes. The plates may be washed twice in 200 μl PBS and incubated, protected from light, in 100 μl of DNA-staining solution (PBS, 0.5 μg/ml Hoechst-33342) for 1 minute. The plates may then be washed once in PBS, and 200 μl PBS finally added per well. The plates, being finally covered with a black adhesive, may be either read directly or stored at 4° C. for later imaging. Where appropriate, the compounds may be added 30 min before stimulation with IFN-γ.
U-2 OS may be seeded at a cell density of 12,000 cells per well in clear-bottom black 96-well Packard View-Plates™. The following day, medium can be removed and replaced with medium containing either the vehicle (DMSO) or increasing concentrations of test compounds for 30 minutes at 37° C. Cells can then be stimulated for 1 hour by adding 10 μl per well of 50 U/ml rhEpo to obtain 5 U/ml of rhEpo as the final concentration. Following treatments, cells can fixed and processed as described above.
STAT-GFP Nuclear Translocation Measurement by Cellomics Automated Fluorescence Microcopy Imaging and Analysis:
The plates can be read on a Cellomics® ArrayscanII automated fluorescence microscope plate reader equipped with a Mercury-Xenon white light illumination source and a Zeiss Axiovert inverted microscope, using the XF100 dichroic/emission filter cube and matching excitation filters, 10× magnification, and a 0.3 numerical aperture objective. Image acquisition and analysis can be performed using a customized protocol based on the ‘NuclearTranslocation’ Bioapplication. For each well, multiple images (fields) can be acquired until a minimum of 1000 cells are counted using two 2 channels: Channel 1 (Hoechst)=focus+nuclear mask, Channel 2 (GFP)=signal quantification in mask areas as outlined below. Nuclei may be first identified based on the Hoechst staining and a mask generated for each nucleus that then serves as a template to generate a circle (eroded inwards by 1 pixel) and a 3 pixel-wide collar-like ring (off-set outwards by 1 pixel), in which the nuclear and cytoplasmic intensity of GFP, respectively, are quantified in the corresponding channel. High content analysis yields numerous measurements per cell and the GFP intensity differential between the nuclear and the cytoplasmic masks may be chosen as a measure of sub-cellular STAT-GFP relocation. The resulting values may be averaged for all cells in the well to return a single measurement plus standard deviation.
To generate IC50 values, the nuclear-cytoplasmic STAT-GFP differential of untreated cells may be used as a baseline and the following equation used to determine the percentage increase in nuclear translocation: Percentage=100*(NCD Compound pre-treated and stimulated−NCD Untreated)/(NCD DMSO-pretreated and stimulated−NCD Untreated). In these assays, compounds of formula (I) generally inhibit JAK2 kinases in the range of 1-10 000 nM.
The activity of the compounds of the formula I can also be determined in vivo:
JAK-2 in vivo
The assay can be performed as described by G. Wernig, T. Mercher, R. Okabe, R. L. Levine, B. H. Lee, D. G. Gilliland, Blood First Edition paper, published online Feb. 14, 2006; DOI 10, 1182/blood-2005-12-4824.
On the basis of these studies, a compound of formula I according to the invention shows therapeutic efficacy especially against disorders dependent on protein kinase, especially proliferative diseases mediated JAK2 kinase activity.
In addition, further protein kinases can be inhibited by compounds of this invention, such as Tyk2, c-src, Flt-3, KDR and others, for each of which test systems are known in the art.
A compound of formula I can be administered alone or in combination with one or more other therapeutic agents, possible combination therapy taking the form of fixed combinations or the administration of a compound of the invention and one or more other therapeutic agents being staggered or given independently of one another, or the combined administration of fixed combinations and one or more other therapeutic agents. A compound of formula I can besides or in addition be administered especially for tumor therapy in combination with chemotherapy, radiotherapy, immunotherapy, surgical intervention, or a combination of these. Long-term therapy is equally possible as is adjuvant therapy in the context of other treatment strategies, as described above. Other possible treatments are therapy to maintain the patient's status after tumor regression, or even chemopreventive therapy, for example in patients at risk.
Thus, a compound of the formula I may be used to advantage in combination with other anti-proliferative compounds. Such antiproliferative compounds include, but are not limited to aromatase inhibitors; antiestrogens; topoisomerase I inhibitors; topoisomerase II inhibitors; microtubule active compounds; alkylating compounds; histone deacetylase inhibitors; compounds which induce cell differentiation processes; cyclooxygenase inhibitors; MMP inhibitors; mTOR inhibitors; antineoplastic antimetabolites; platin compounds; compounds targeting/decreasing a protein or lipid kinase activity and further anti-angiogenic compounds; compounds which target, decrease or inhibit the activity of a protein or lipid phosphatase; gonadorelin agonists; anti-androgens; methionine aminopeptidase inhibitors; bisphosphonates; biological response modifiers; antiproliferative antibodies; heparanase inhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasome inhibitors; compounds used in the treatment of hematologic malignancies; compounds which target, decrease or inhibit the activity of Flt-3; Hsp90 inhibitors such as 17-AAG (17-allylaminogeldanamycin, NSC330507), 17-DMAG (17-dimethylaminoethylamino-17-demethoxy-geldanamycin, NSC707545), IPI-504, CNF1010, CNF2024, CNF1010 from Conforma Therapeutics; temozolomide (TEMODAL®); kinesin spindle protein inhibitors, such as SB715992 or SB743921 from GlaxoSmithKline, or pentamidine/chlorpromazine from CombinatoRx; MEK inhibitors such as ARRY142886 from Array PioPharma, AZD6244 from AstraZeneca, PD181461 from Pfizer, leucovorin, EDG binders, antileukemia compounds, ribonucleotide reductase inhibitors, S-adenosylmethionine decarboxylase inhibitors, antiproliferative antibodies or other chemotherapeutic compounds. Further, alternatively or in addition they may be used in combination with other tumor treatment approaches, including surgery, ionizing radiation, photodynamic therapy, implants, e.g. with corticosteroids, hormones, or they may be used as radiosensitizers. Also, in anti-inflammatory and/or antiproliferative treatment, combination with anti-inflammatory drugs is included. Combination is also possible with antihistamine drug substances, bronchodilatatory drugs, NSAID or antagonists of chemokine receptors.
The term “aromatase inhibitor” as used herein relates to a compound which inhibits the estrogen production, i.e. the conversion of the substrates androstenedione and testosterone to estrone and estradiol, respectively. The term includes, but is not limited to steroids, especially atamestane, exemestane and formestane and, in particular, non-steroids, especially aminoglutethimide, roglethimide, pyridoglutethimide, trilostane, testolactone, ketokonazole, vorozole, fadrozole, anastrozole and letrozole. Exemestane can be administered, e.g., in the form as it is marketed, e.g. under the trademark AROMASIN. Formestane can be administered, e.g., in the form as it is marketed, e.g. under the trademark LENTARON. Fadrozole can be administered, e.g., in the form as it is marketed, e.g. under the trademark AFEMA. Anastrozole can be administered, e.g., in the form as it is marketed, e.g. under the trademark ARIMIDEX. Letrozole can be administered, e.g., in the form as it is marketed, e.g. under the trademark FEMARA or FEMAR. Aminoglutethimide can be administered, e.g., in the form as it is marketed, e.g. under the trademark ORIMETEN. A combination of the invention comprising a chemotherapeutic agent which is an aromatase inhibitor is particularly useful for the treatment of hormone receptor positive tumors, e.g. breast tumors.
The term “antiestrogen” as used herein relates to a compound which antagonizes the effect of estrogens at the estrogen receptor level. The term includes, but is not limited to tam-oxifen, fulvestrant, raloxifene and raloxifene hydrochloride. Tamoxifen can be administered, e.g., in the form as it is marketed, e.g. under the trademark NOLVADEX. Raloxifene hydro-chloride can be administered, e.g., in the form as it is marketed, e.g. under the trademark EVISTA. Fulvestrant can be formulated as disclosed in U.S. Pat. No. 4,659,516 or it can be administered, e.g., in the form as it is marketed, e.g. under the trademark FASLODEX. A combination of the invention comprising a chemotherapeutic agent which is an antiestrogen is particularly useful for the treatment of estrogen receptor positive tumors, e.g. breast tumors.
The term “anti-androgen” as used herein relates to any substance which is capable of inhibiting the biological effects of androgenic hormones and includes, but is not limited to, bicalutamide (CASODEX), which can be formulated, e.g. as disclosed in U.S. Pat. No. 4,636,505. The term “gonadorelin agonist” as used herein includes, but is not limited to abarelix, goserelin and goserelin acetate. Goserelin is disclosed in U.S. Pat. No. 4,100,274 and can be administered, e.g., in the form as it is marketed, e.g. under the trademark ZOLADEX. Abarelix can be formulated, e.g. as disclosed in U.S. Pat. No. 5,843,901.
The term “topoisomerase I inhibitor” as used herein includes, but is not limited to topotecan, gimatecan, irinotecan, camptothecian and its analogues, 9-nitrocamptothecin and the macromolecular camptothecin conjugate PNU-166148 (compound A1 in WO99/17804). Irinotecan can be administered, e.g. in the form as it is marketed, e.g. under the trademark CAMPTOSAR. Topotecan can be administered, e.g., in the form as it is marketed, e.g. under the trademark HYCAMTIN.
The term “topoisomerase II inhibitor” as used herein includes, but is not limited to the anthracyclines such as doxorubicin (including liposomal formulation, e.g. CAELYX), daunorubicin, epirubicin, idarubicin and nemorubicin, the anthraquinones mitoxantrone and losoxantrone, and the podophillotoxines etoposide and teniposide. Etoposide can be administered, e.g. in the form as it is marketed, e.g. under the trademark ETOPOPHOS. Teniposide can be administered, e.g. in the form as it is marketed, e.g. under the trademark VM 26-BRISTOL. Doxorubicin can be administered, e.g. in the form as it is marketed, e.g. under the trademark ADRIBLASTIN or ADRIAMYCIN. Epirubicin can be administered, e.g. in the form as it is marketed, e.g. under the trademark FARMORUBICIN. Idarubicin can be administered, e.g. in the form as it is marketed, e.g. under the trademark ZAVEDOS. Mitoxantrone can be administered, e.g. in the form as it is marketed, e.g. under the trademark NOVANTRON.
The term “microtubule active compound” relates to microtubule stabilizing, microtubule destabilizing compounds and microtublin polymerization inhibitors including, but not limited to taxanes, e.g. paclitaxel and docetaxel, vinca alkaloids, e.g., vinblastine, especially vinblastine sulfate, vincristine especially vincristine sulfate, and vinorelbine, discodermolides, cochicine and epothilones and derivatives thereof, e.g. epothilone B or D or derivatives thereof. Paclitaxel may be administered e.g. in the form as it is marketed, e.g. TAXOL.
Docetaxel can be administered, e.g., in the form as it is marketed, e.g. under the trademark TAXOTERE. Vinblastine sulfate can be administered, e.g., in the form as it is marketed, e.g. under the trademark VINBLASTIN R.P. Vincristine sulfate can be administered, e.g., in the form as it is marketed, e.g. under the trademark FARMISTIN. Discodermolide can be obtained, e.g., as disclosed in U.S. Pat. No. 5,010,099. Also included are Epothilone derivatives which are disclosed in WO 98/10121, U.S. Pat. No. 6,194,181, WO 98/25929, WO 98/08849, WO 99/43653, WO 98/22461 and WO 00/31247. Especially preferred are Epothilone A and/or B.
The term “alkylating compound” as used herein includes, but is not limited to, cyclophosphamide, ifosfamide, melphalan or nitrosourea (BCNU or Gliadel). Cyclophosphamide can be administered, e.g., in the form as it is marketed, e.g. under the trademark CYCLOSTIN. Ifosfamide can be administered, e.g., in the form as it is marketed, e.g. under the trademark HOLOXAN.
The term “histone deacetylase inhibitors” or “HDAC inhibitors” relates to compounds which inhibit the histone deacetylase and which possess antiproliferative activity. This includes compounds disclosed in WO 02/22577, especially N-hydroxy-3-[4-[[(2-hydroxyethyl)[2-(1H-indol-3-yl)ethyl]-amino]methyl]phenyl]-2E -2-propenamide, N-hydroxy-3-[4-[[[2-(2-methyl-1H-indol-3-yl)-ethyl]-amino]methyl]phenyl]-2E-2-propenamide and pharmaceutically acceptable salts thereof. It further especially includes Suberoylanilide hydroxamic acid (SAHA).
The term “antineoplastic antimetabolite” includes, but is not limited to, 5-Fluorouracil or 5-FU, capecitabine, gemcitabine, DNA demethylating compounds, such as 5-azacytidine and decitabine, methotrexate and edatrexate, and folic acid antagonists such as pemetrexed. Capecitabine can be administered, e.g., in the form as it is marketed, e.g. under the trademark XELODA. Gemcitabine can be administered, e.g., in the form as it is marketed, e.g. under the trademark GEMZAR.
The term “platin compound” as used herein includes, but is not limited to, carboplatin, cisplatin, cisplatinum and oxaliplatin. Carboplatin can be administered, e.g., in the form as it is marketed, e.g. under the trademark CARBOPLAT. Oxaliplatin can be administered, e.g., in the form as it is marketed, e.g. under the trademark ELOXATIN.
The term “compounds targeting/decreasing a protein or lipid kinase activity”; or a “protein or lipid phosphatase activity”; or “further anti-angiogenic compounds” as used herein includes, but is not limited to, protein tyrosine kinase and/or serine and/or threonine kinase inhibitors or lipid kinase inhibitors, e.g.,
Further anti-angiogenic compounds include compounds having another mechanism for their activity, e.g. unrelated to protein or lipid kinase inhibition e.g. thalidomide (THALOMID) and TNP-470.
The term “Compounds which target, decrease or inhibit the activity of a protein or lipid phosphatase” includes, but is not limited to inhibitors of phosphatase 1, phosphatase 2A, or CDC25, e.g. okadaic acid or a derivative thereof.
The term “Compounds which induce cell differentiation processes” includes, but is not limited to e.g. retinoic acid, α- γ- or ε-tocopherol or α- γ- or δ-tocotrienol.
The term “cyclooxygenase inhibitor” as used herein includes, but is not limited to, e.g. Cox-2 inhibitors, 5-alkyl substituted 2-arylaminophenylacetic acid and derivatives, such as celecoxib (CELEBREX), rofecoxib (VIOXX), etoricoxib, valdecoxib or a 5-alkyl-2-arylaminophenylacetic acid, e.g. 5-methyl-2-(2′-chloro-6′-fluoroanilino)phenyl acetic acid, lumiracoxib.
The term “bisphosphonates” as used herein includes, but is not limited to, etridonic, clodronic, tiludronic, pamidronic, alendronic, ibandronic, risedronic and zoledronic acid. “Etridonic acid” can be administered, e.g., in the form as it is marketed, e.g. under the trademark DIDRONEL. “Clodronic acid” can be administered, e.g., in the form as it is marketed, e.g. under the trademark BONEFOS. “Tiludronic acid” can be administered, e.g., in the form as it is marketed, e.g. under the trademark SKELID. “Pamidronic acid” can be administered, e.g. in the form as it is marketed, e.g. under the trademark AREDIA™. “Alendronic acid” can be administered, e.g., in the form as it is marketed, e.g. under the trademark FOSAMAX. “Ibandronic acid” can be administered, e.g., in the form as it is marketed, e.g. under the trademark BONDRANAT. “Risedronic acid” can be administered, e.g., in the form as it is marketed, e.g. under the trademark ACTONEL. “Zoledronic acid” can be administered, e.g. in the form as it is marketed, e.g. under the trademark ZOMETA.
The term “mTOR inhibitors” relates to compounds which inhibit the mammalian target of rapamycin (mTOR) and which possess antiproliferative activity such as sirolimus (Rapamune®), everolimus (Certican™), CCI-779 and ABT578.
The term “heparanase inhibitor” as used herein refers to compounds which target, decrease or inhibit heparin sulfate degradation. The term includes, but is not limited to, PI-88. The term “biological response modifier” as used herein refers to a lymphokine or interferons, e.g. interferon γ.
The term “inhibitor of Ras oncogenic isoforms”, e.g. H-Ras, K-Ras, or N-Ras, as used herein refers to compounds which target, decrease or inhibit the oncogenic activity of Ras e.g. a “farnesyl transferase inhibitor” e.g. L-744832, DK8G557 or R115777 (Zarnestra). The term “telomerase inhibitor” as used herein refers to compounds which target, decrease or inhibit the activity of telomerase. Compounds which target, decrease or inhibit the activity of telomerase are especially compounds which inhibit the telomerase receptor, e.g. telomestatin.
The term “methionine aminopeptidase inhibitor” as used herein refers to compounds which target, decrease or inhibit the activity of methionine aminopeptidase. Compounds which target, decrease or inhibit the activity of methionine aminopeptidase are e.g. bengamide or a derivative thereof.
The term “proteasome inhibitor” as used herein refers to compounds which target, decrease or inhibit the activity of the proteasome. Compounds which target, decrease or inhibit the activity of the proteasome include e.g. Bortezomid (Velcade™) and MLN 341.
The term “matrix metalloproteinase inhibitor” or (“MMP” inhibitor) as used herein includes, but is not limited to, collagen peptidomimetic and nonpeptidomimetic inhibitors, tetracycline derivatives, e.g. hydroxamate peptidomimetic inhibitor batimastat and its orally bioavailable analogue marimastat (BB-2516), prinomastat (AG3340), metastat (NSC 683551) BMS-279251, BAY 12-9566, TAA211, MMI270B or AAJ996.
The term “compounds used in the treatment of hematologic malignancies” as used herein includes, but is not limited to, FMS-like tyrosine kinase inhibitors e.g. compounds targeting, decreasing or inhibiting the activity of FMS-like tyrosine kinase receptors (Flt-3R); interferon, 1-b-D-arabinofuransylcytosine (ara-c) and bisulfan; and ALK inhibitors e.g. compounds which target, decrease or inhibit anaplastic lymphoma kinase.
Compounds which target, decrease or inhibit the activity of FMS-like tyrosine kinase receptors (Flt-3R) are especially compounds, proteins or antibodies which inhibit members of the Flt-3R receptor kinase family, e.g. PKC412, midostaurin, a staurosporine derivative, SU11248 and MLN518.
The term “HSP90 inhibitors” as used herein includes, but is not limited to, compounds targeting, decreasing or inhibiting the intrinsic ATPase activity of HSP90; degrading, targeting, decreasing or inhibiting the HSP90 client proteins via the ubiquitin proteosome pathway. Compounds targeting, decreasing or inhibiting the intrinsic ATPase activity of HSP90 are especially compounds, proteins or antibodies which inhibit the ATPase activity of HSP90 e.g., 17-allylamino,17-demethoxygeldanamycin (17AAG), a geldanamycin derivative; other geldanamycin related compounds; radicicol and HDAC inhibitors.
The term “antiproliferative antibodies” as used herein includes, but is not limited to, trastuzumab (Herceptin™), Trastuzumab-DCM1,erbitux, bevacizumab (Avastin™), rituximab (Rituxan®), PRO64553 (anti-CD40) and 2C4 Antibody. By antibodies is meant e.g. intact monoclonal antibodies, polyclonal antibodies, multispecific antibodies formed from at least 2 intact antibodies, and antibodies fragments so long as they exhibit the desired biological activity.
For the treatment of acute myeloid leukemia (AML), compounds of formula (I) can be used in combination with standard leukemia therapies, especially in combination with therapies used for the treatment of AML. In particular, compounds of formula (I) can be administered in combination with, e.g., farnesyl transferase inhibitors and/or other drugs useful for the treatment of AML, such as Daunorubicin, Adriamycin, Ara-C, VP-16, Teniposide, Mitoxantrone, Idarubicin, Carboplatinum and PKC412.
The term “antileukemic compounds” includes, for example, Ara-C, a pyrimidine analog, which is the 2′-alpha-hydroxy ribose (arabinoside) derivative of deoxycytidine. Also included is the purine analog of hypoxanthine, 6-mercaptopurine (6-MP) and fludarabine phosphate. Compounds which target, decrease or inhibit activity of histone deacetylase (HDAC) inhibitors such as sodium butyrate and suberoylanilide hydroxamic acid (SAHA) inhibit the activity of the enzymes known as histone deacetylases. Specific HDAC inhibitors include MS275, SAHA, FK228 (formerly FR901228), Trichostatin A and compounds disclosed in U.S. Pat. No. 6,552,065, in particular, N-hydroxy-3-[4-[[[2-(2-methyl-1H-indol-3-yl)-ethyl]-amino]methyl]phenyl]-2E-2-propenamide, or a pharmaceutically acceptable salt thereof and N-hydroxy-3-[4-[(2-hydroxyethyl){2-(1H-indol-3-yl)ethyl]-amino]methyl]phenyl]-2E-2-propeneamide, or a pharmaceutically acceptable salt thereof, especially the lactate salt.
“Somatostatin receptor antagonists” as used herein refers to compounds which target, treat or inhibit the somatostatin receptor such as octreotide, and SOM230.
“Tumor cell damaging approaches” refer to approaches such as ionizing radiation. The term “ionizing radiation” referred to above and hereinafter means ionizing radiation that occurs as either electromagnetic rays (such as X-rays and gamma rays) or particles (such as alpha and beta particles). Ionizing radiation is provided in, but not limited to, radiation therapy and is known in the art. See Hellman, Principles of Radiation Therapy, Cancer, in Principles and Practice of Oncology, Devita et al., Eds., 4th Edition, Vol. 1, pp. 248-275 (1993).
The term “EDG binders” as used herein refers a class of immunosuppressants that modulates lymphocyte recirculation, such as FTY720.
The term “ribonucleotide reductase inhibitors” includes, but is not limited to to pyrimidine or purine nucleoside analogs including, but not limited to, fludarabine and/or cytosine arabinoside (ara-C), 6-thioguanine, 5-fluorouracil, cladribine, 6-mercaptopurine (especially in combination with ara-C against ALL) and/or pentostatin. Ribonucleotide reductase inhibitors are especially hydroxyurea or 2-hydroxy-1H-isoindole-1,3-dione derivatives, such as PL-1, PL-2, PL-3, PL-4, PL-5, PL-6, PL-7 or PL-8 mentioned in Nandy et al., Acta Oncologica, Vol. 33, No. 8, pp. 953-961 (1994).
The term “S-adenosylmethionine decarboxylase inhibitors” as used herein includes, but is not limited to the compounds disclosed in U.S. Pat. No. 5,461,076.
Also included are in particular those compounds, proteins or monoclonal antibodies of VEGF disclosed in WO 98/35958, e.g. 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine or a pharmaceutically acceptable salt thereof, e.g. the succinate, or in WO 00/09495, WO 00/27820, WO 00/59509, WO 98/11223, WO 00/27819 and EP 0 769 947; those as described by Prewett et al, Cancer Res, Vol. 59, pp. 5209-5218 (1999); Yuan et al., Proc Natl Acad Sci USA, Vol. 93, pp. 14765-14770 (1996); Zhu et al., Cancer Res, Vol. 58, pp. 3209-3214 (1998); and Mordenti et al., Toxicol Pathol, Vol. 27, No. 1, pp. 14-21 (1999); in WO 00/37502 and WO 94/10202; ANGIOSTATIN, described by O'Reilly et al., Cell, Vol. 79, pp. 315-328 (1994); ENDOSTATIN, described by O'Reilly et al., Cell, Vol. 88, pp. 277-285 (1997); anthranilic acid amides; ZD4190; ZD6474; SU5416; SU6668; bevacizumab; or anti-VEGF antibodies or anti-VEGF receptor antibodies, e.g. rhuMAb and RHUFab, VEGF aptamer e.g. Macugon; FLT-4 inhibitors, FLT-3 inhibitors, VEGFR-2 IgG1 antibody, Angiozyme (RPI 4610) and Bevacizumab (Avastin™).
“Photodynamic therapy” as used herein refers to therapy which uses certain chemicals known as photosensitizing compounds to treat or prevent cancers. Examples of photodynamic therapy includes treatment with compounds, such as e.g. VISUDYNE and porfimer sodium.
“Angiostatic steroids” as used herein refers to compounds which block or inhibit angiogenesis, such as, e.g., anecortave, triamcinolone. hydrocortisone, 11-α-epihydrocotisol, cortexolone, 17α-hydroxyprogesterone, corticosterone, desoxycorticosterone, testosterone, estrone and dexamethasone.
“Implants containing corticosteroids” as used herein includes, but is not limited to compounds, such as e.g. fluocinolone, dexamethasone.
“Other chemotherapeutic compounds” include, but are not limited to, plant alkaloids, hormonal compounds and antagonists; biological response modifiers, preferably lymphokines or interferons; antisense oligonucleotides or oligonucleotide derivatives; shRNA or siRNA; or miscellaneous compounds or compounds with other or unknown mechanism of action.
The compounds of the invention are also useful as co-therapeutic compounds for use in combination with other drug substances such as anti-inflammatory, bronchodilatory or antihistamine drug substances, particularly in the treatment of inflammatory diseases such as those mentioned hereinbefore, for example as potentiators of therapeutic activity of such drugs or as a means of reducing required dosaging or potential side effects of such drugs. A compound of the invention may be mixed with the other drug substance in a fixed pharmaceutical composition or it may be administered separately, before, simultaneously with or after the other drug substance. Accordingly the invention includes a combination of a compound of the invention as hereinbefore described with an anti-inflammator or antihistamine drug substance, said compound of the invention and said drug substance being in the same or different pharmaceutical composition.
Suitable anti-inflammatory drugs include steroids, in particular glucocorticosteroids such as budesonide, beclamethasone dipropionate, fluticasone propionate, ciclesonide or mometasone furoate, or steroids described in WO 02/88167, WO 02/12266, WO 02/100879, WO 02/00679 (especially those of Examples 3, 11, 14, 17, 19, 26, 34, 37, 39, 51, 60, 67, 72, 73, 90, 99 and 101), WO 03/035668, WO 03/048181, WO 03/062259, WO 03/064445, WO 03/072592, non-steroidal glucocorticoid receptor agonists such as those described in WO 00/00531, WO 02/10143, WO 03/082280, WO 03/082787, WO 03/104195, WO 04/005229; LTB4 antagonists such LY293111, CGSO25019C, CP-195543, SC-53228, BIIL 284, ONO 4057, SB 209247 and those described in U.S. Pat. No. 5,451,700; LTD4 antagonists such as montelukast and zafirlukast; PDE4 inhibitors such cilomilast (Ariflo® GlaxoSmithKline), Roflumilast (Byk Gulden),V-11294A (Napp), BAY19-8004 (Bayer), SCH-351591 (Schering-Plough), Arofylline (Almirall Prodesfarma), PD189659/PD168787 (Parke-Davis), AWD-12-281 (Asta Medica), CDC-801 (Celgene), SeICID™ CC-10004 (Celgene), VM554/UM565 (Vernalis), T-440 (Tanabe), KW-4490 (Kyowa Hakko Kogyo), and those disclosed in WO 92/19594, WO 93/19749, WO 93/19750, WO 93/19751, WO 98/18796, WO 99/16766, WO 01/13953, WO 03/104204, WO 03/104205, WO 03/39544, WO 04/000814, WO 04/000839, WO 04/005258, WO 04/018450, WO 04/018451, WO 04/018457, WO 04/018465, WO 04/018431, WO 04/018449, WO 04/018450, WO 04/018451, WO 04/018457, WO 04/018465, WO 04/019944, WO 04/019945, WO 04/045607 and WO 04/037805; A2a agonists such as those disclosed in EP 409595A2, EP 1052264, EP 1241176, WO 94/17090, WO 96/02543, WO 96/02553, WO 98/28319, WO 99/24449, WO 99/24450, WO 99/24451, WO 99/38877, WO 99/41267, WO 99/67263, WO 99/67264, WO 99/67265, WO 99/67266, WO 00/23457, WO 00/77018, WO 00/78774, WO 01/23399, WO 01/27130, WO 01/27131, WO 01/60835, WO 01/94368, WO 02/00676, WO 02/22630, WO 02/96462, WO 03/086408, WO 04/ 039762, WO 04/039766, WO 04/045618 and WO 04/046083; A2b antagonists such as those described in WO 02/42298; and beta-2 adrenoceptor agonists such as albuterol (salbutamol), metaproterenol, terbutaline, salmeterol fenoterol, procaterol, and especially, formoterol and pharmaceutically acceptable salts thereof, and compounds (in free or salt or solvate form) of formula I of WO 0075114, which document is incorporated herein by reference, preferably compounds of the Examples thereof, especially a compound of formula
and pharmaceutically acceptable salts thereof, as well as compounds (in free or salt or solvate form) of formula I of WO 04/16601, and also compounds of WO 04/033412. Suitable bronchodilatory drugs include anticholinergic or antimuscarinic compounds, in particular ipratropium bromide, oxitropium bromide, tiotropium salts and CHF 4226 (Chiesi), and glycopyrrolate, but also those described in WO 01/04118, WO 02/51841, WO 02/53564, WO 03/00840, WO 03/87094, WO 04/05285, WO 02/00652, WO 03/53966, EP 424021, U.S. Pat. Nos. 5,171,744, 3,714,357, WO 03/33495 and WO 04/018422.
Suitable antihistamine drug substances include cetirizine hydrochloride, acetaminophen, clemastine fumarate, promethazine, loratidine, desloratidine, diphenhydramine and fexofenadine hydrochloride, activastine, astemizole, azelastine, ebastine, epinastine, mizolastine and tefenadine as well as those disclosed in WO 03/099807, WO 04/026841 and JP 2004107299.
Other useful combinations of compounds of the invention with anti-inflammatory drugs are those with antagonists of chemokine receptors, e.g. CCR-1, CCR-2, CCR-3, CCR-4, CCR-5, CCR-6, CCR-7, CCR-8, CCR-9 and CCR10, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, particularly CCR-5 antagonists such as Schering-Plough antagonists SC-351125, SCH-55700 and SCH-D, Takeda antagonists such as N-[[4-[[[6,7-dihydro-2-(4-methylphenyl)-5H-benzo-cyclohepten-8-yl]carbonyl]amino]phenyl]-methyl]tetrahydro-N,N-dimethyl-2H-pyran-4-amin-ium chloride (TAK-770), and CCR-5 antagonists described in U.S. Pat. No. 6,166,037 (particularly claims 18 and 19), WO 00/66558 (particularly claim 8), WO 00/66559 (particularly claim 9), WO 04/018425 and WO 04/026873.
Therapeutic agents for possible combination are especially one or more antiproliferative, cytostatic or cytotoxic compounds, for example one or several agents selected from the group which includes, but is not limited to, an inhibitor of polyamine biosynthesis, an inhibitor of a protein kinase, especially of a serine/threonine protein kinase, such as protein kinase C, or of a tyrosine protein kinase, such as the EGF receptor tyrosine kinase, e.g. Iressa®, the VEGF receptor tyrosine kinase, e.g. PTK787 or Avastin®, or the PDGF receptor tyrosine kinase, e.g. STI571 (Glivec®), a cytokine, a negative growth regulator, such as TGF-β or IFN-β, an aromatase inhibitor, e.g. letrozole (Femara®) or anastrozole, an inhibitor of the interaction of an SH2 domain with a phosphorylated protein, antiestrogens, topoisomerase I inhibitors, such as irinotecan, topoisomerase II inhibitors, microtubule active agents, e.g. paclitaxel or an epothilone, alkylating agents, antiproliferative antimetabolites, such as gemcitabine or capecitabine, platin compounds, such as carboplatin or cis-platin, bisphosphonates, e.g. AREDIA® or ZOMETA®, and monoclonal antibodies, e.g. against HER2, such as trastuzumab.
The structure of the active agents identified by code nos., generic or trade names may be taken from the actual edition of the standard compendium “The Merck Index” or from databases, e.g. Patents International (e.g. IMS World Publications). The corresponding content thereof is hereby incorporated by reference.
The above-mentioned compounds, which can be used in combination with a compound of the formula I, can be prepared and administered as described in the art, such as in the documents cited above.
Thus, the invention relates in a further aspect to a combination comprising a therapeutically effective amount of a compound of formula I in free form or in pharmaceutically acceptable salt form and a second drug substance, for simultaneous or sequential administration.
The invention also provides, in a further aspect, a pharmaceutical preparation (composition), comprising a compound of formula I as defined herein, or a pharmaceutically acceptable salt of such a compound, or a hydrate or solvate thereof, and at least one pharmaceutically acceptable carrier and/or diluents and optionally one or more further drug substances.
The compounds of the invention may be administered by any conventional route, in particular parenterally, for example in the form of injectable solutions or suspensions, enterally, e.g. orally, for example in the form of tablets or capsules, topically, e.g. in the form of lotions, gels, ointments or creams, or in a nasal or a suppository form. Topical administration is e.g. to the skin. A further form of topical administration is to the eye. Pharmaceutical compositions comprising a compound of the invention in association with at least one pharmaceutical acceptable carrier or diluent may be manufactured in conventional manner by mixing with a pharmaceutically acceptable carrier or diluent.
The invention relates also to pharmaceutical compositions comprising an effective amount, especially an amount effective in the treatment of one of the above-mentioned diseases (=disorders), of a compound of formula I or a pharmaceutically acceptable salt thereof together with one or more pharmaceutically acceptable carriers that are suitable for topical, enteral, for example oral or rectal, or parenteral administration and that may be inorganic or organic, solid or liquid. There can be used for oral administration especially tablets or gelatin capsules that comprise the active ingredient together with diluents, for example lactose, dextrose, mannitol, and/or glycerol, and/or lubricants and/or polyethylene glycol. Tablets may also comprise binders, for example magnesium aluminum silicate, starches, such as corn, wheat or rice starch, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone, and, if desired, disintegrators, for example starches, agar, alginic acid or a salt thereof, such as sodium alginate, and/or effervescent mixtures, or adsorbents, dyes, flavorings and sweeteners. It is also possible to use the pharmacologically active compounds of the present invention in the form of parenterally administrable compositions or in the form of infusion solutions. The pharmaceutical compositions may be sterilized and/or may comprise excipients, for example preservatives, stabilisers, wetting compounds and/or emulsifiers, solubilisers, salts for regulating the osmotic pressure and/or buffers. The present pharmaceutical compositions, which may, if desired, comprise other pharmacologically active substances are prepared in a manner known per se, for example by means of conventional mixing, granulating, confectionning, dissolving or lyophilising processes, and comprise approximately from 1% to 99%, especially from approx. 1% to approx. 20%, active ingredient(s).
The dosage of the active ingredient to be applied to a warm-blooded animal depends upon a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound employed. A physician, clinician or veterinarian of ordinary skill can readily determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progress of the condition. Optimal precision in achieving concentration of drug within the range that yields efficacy without toxicity requires a regimen based on the kinetics of the drug's availability to target sites. This involves a consideration of the distribution, equilibrium, and elimination of a drug. The dose of a compound of the formula I or a pharmaceutically acceptable salt thereof to be administered to warm-blooded animals, for example humans of approximately 70 kg body weight, is preferably from approximately 3 mg to approximately 5 g, more preferably from approximately 10 mg to approximately 1.5 g per person per day, divided preferably into 1 to 3 single doses which may, for example, be of the same size. Usually, children receive half of the adult dose.
In a further aspect, the invention relates to a compound of formula I or a pharmaceutically acceptable salt, as a medicament/for use as a medicament, in particular for the treatment of one or more Protein tyrosine kinase mediated diseases.
In a further aspect, the invention relates to the use of a compound of formula I or a pharmaceutically acceptable salt,as active ingredient in a medicament, in particular for the treatment of one or more Protein tyrosine kinase mediated diseases.
In a further aspect, the invention relates to the use of a compound of formula I or a pharmaceutically acceptable salt, as medicament, in particular for the treatment of one or more Protein tyrosine kinase mediated diseases.
In a further aspect, the invention relates to the use of a compound of formula I or a pharmaceutically acceptable salt, for the manufacture of a medicament for the treatment of one or more Protein tyrosine kinase mediated diseases.
In a further aspect, the invention relates to a compound of formula I or a pharmaceutically acceptable salt of such a compound, for use in a method for the treatment of a subject in need thereof, especially for the treatment of a Protein tyrosine kinase mediated disease, most especially in a patient requiring such treatment.
In a further aspect, the invention relates to a method for the treatment of a disease which responds to an inhibition of JAK-2 and/or Jak-3 kinase, which comprises administering a compound of formula I or a pharmaceutically acceptable salt thereof, wherein the radicals and symbols have the meanings as defined above, especially in a quantity effective against said disease, to a warm-blooded animal requiring such treatment.
In a further aspect, the invention relates to a pharmaceutical composition comprising a compound of formula I as active ingredient in association with at least one pharmaceutical carrier or diluent. Such compositions may be manufactured in conventional manner. In a further aspect, the invention relates to a method of treatment of one or more Protein tyrosine kinase mediated diseases, in a subject in need of such treatment, which comprises administering to such subject a therapeutically effective amount of compound of formula I.
In a further aspect, the invention relates to pharmaceutical compositions comprising: (a) an effective amount of compound of formula I and pharmaceutically acceptable salts, pharmaceutically acceptable prodrugs, and pharmaceutically active metabolites thereof; and (b) one or more pharmaceutically acceptable excipients and/or diluents.
In a further aspect, the invention relates to a pharmaceutical composition for treatment of a disease, e.g. of solid or liquid tumours in warm-blooded animals, including humans, comprising a dose effective in the treatment of said disease of a compound of the formula I as described above or a pharmaceutically acceptable salt of such a compound together with a pharmaceutically acceptable carrier (=carrier material).
A compound of the formula I or I′ may be prepared by processes that, though not applied hitherto for the new compounds of the present invention where they thus form new processes, are known per se, the following scheme illustrates methods for such preparation.
Preferably, a process for the manufacture of a compound of the formula I comprises either
wherein the substituents are as defined for a compound of the formula I and LG represents a leaving group (such as tosylate, mesylate or halo, in particular chloride) with a compound of the formula IV or IV′,
wherein the substituents are as defined for a compound of the formula I and LG represents a leaving group (such as tosylate, mesylate or halo, in particular bromide) in a Cu-catalyzed Buchwald reaction to obtain a compound of formula (VI) or (VI′)
wherein the substituents are as defined above with a compound of formula I and reacting in a second step the obtained compound of formula VI with a compound of formula (III)
NH2—R1 (III)
wherein the substituents are as defined for a compound of the formula I and LG represents a leaving group (such as tosylate, mesylate or halo, in particular chloride) with a compound of the formula III,
NH2—R1 (III)
wherein the substituents are as defined above for a compound of formula I and reacting in a second step the obtained compound of formula V with a compound of formula IV
Where temperatures are given hereinbefore or hereinafter, “about” has to be added, as minor deviations from the numeric values given, e.g. variations of ±10%, are tolerable. All reactions may take place in the presence of one or more diluents and/or solvents. The starting materials may be used in equimolar amounts; alternatively, a compound may be used in excess, e.g. to function as a solvent or to shift equilibrium or to generally accelerate reaction rates. Reaction aids, such as acids, bases or catalysts may be added in suitable amounts, as known in the field, required by a reaction and in line with generally known procedures.
Buchwald Reaction
This reaction, also known as Buchwald amination or Buchwald-Hartwig reaction is generally known in the field. This reaction is catalyzed by transition metals, in particular Cu or Pd complexes or salts; takes place in the presence of one or more basic compounds (such as an amine or an alkalialkoxide) and one or more diluents (such as polar aprotic diluents). Further details may be found in the examples.
Protecting Groups
If one or more other functional groups, for example carboxy, hydroxy, amino, sulfhydryl or the like are or need to be protected in a starting material as described herein or any other precursor, because they should not take part in the reaction or disturb the reaction, these are such groups as are usually used in the synthesis of peptide compounds, and also of cephalosporins and penicillins, as well as nucleic acid derivatives and sugars. Protecting groups are such groups that are no longer present in the final compounds once they are removed, while groups that remain as substituents are not protecting groups in the sense used here which are groups that are added at a starting material or intermediate stage and removed to obtain a final compound. Also in the case of conversions of a compound of the formula I into a different compound of the formula I, protecting groups may be introduced and removed, if useful or required.
The protecting groups may already be present in precursors and should protect the functional groups concerned against unwanted secondary reactions, such as acylations, etherifications, esterifications, oxidations, solvolysis, and similar reactions. It is a characteristic of protecting groups that they lend themselves readily, i.e. without undesired secondary reactions, to removal, typically by acetolysis, protonolysis, solvolysis, reduction, photolysis or also by enzyme activity, for example under conditions analogous to physiological conditions, and that they are not present in the end-products. The specialist knows, or can easily establish, which protecting groups are suitable with the reactions mentioned above and below.
The protection of such functional groups by such protecting groups, the protecting groups themselves, and their removal reactions are described for example in standard reference works, such as J. F. W. McOmie, “Protective Groups in Organic Chemistry”, Plenum Press, London and New York 1973, in T. W. Greene, “Protective Groups in Organic Synthesis”, Third edition, Wiley, New York 1999, in “The Peptides”; Volume 3 (editors: E. Gross and J. Meienhofer), Academic Press, London and New York 1981, in “Methoden der organischen Chemie” (Methods of organic chemistry), Houben Weyl, 4th edition, Volume 15/I, Georg Thieme Verlag, Stuttgart 1974, in H.-D. Jakubke and H. Jescheit, “Aminosäuren, Peptide, Proteine” (Amino acids, peptides, proteins), Verlag Chemie, Weinheim, Deerfield Beach, and Basel 1982, and in Jochen Lehmann, “Chemie der Kohlenhydrate: Monosaccharide und Derivate” (Chemistry of carbohydrates: monosaccharides and derivatives), Georg Thieme Verlag, Stuttgart 1974.
Optional Reactions and Conversions
A compound of the formula I may be converted into a different compound of the formula I.
For example, in a compound of the formula I wherein R1 or especially R4 carries an amino or amino-C1-C7-alkyl substituent, the amino can be converted into acylamino, e.g. C1-C7-alkanoylamino or C1-C7-alkanesulfonylamino, by reaction with a corresponding C1-C7-alkanoylhalogenide or C1-C7-alkanesulfonylhalogenide, e.g. a corresponding chloride, in the presence of a tertiary nitrogen base, such as triethylamine or pyridine, in the absence or presence of an appropriate solvent, such a methylene chloride, for example at temperatures in the range from −20 to 50° C., e.g. at about room temperature.
In a compound of the formula I wherein R1 or especially R4 carries a cyano substituent, the cyano may be converted to an aminomethyl group, e.g. by hydrogenation in the presence of an appropriate metal catalyst, such as Raney Nickel or Raney Cobalt, in an appropriate solvent, e.g. a lower alkanol, such as methanol and/or ethanol, for example at temperatures in the range from −20 to 50° C., e.g. at about room temperature.
In a compound of the formula I wherein R1 or especially R4 carries a carboxyl (COOH) substituent, the latter can be converted into an amide group, e.g. an N—C1-C7-alkyl-carbamoyl group, by reaction with the corresponding amine, e.g. in the presence of a coupling agent, that forms a preferred reactive derivative of the carboxyl group in situ, for example dicyclohexylcarbodiimide/1-hydroxybenzotriazole (DCC/HOBT); bis(2-oxo-3-oxazolidinyl)phosphinic chloride (BOPCl); O-(1,2-dihydro-2-oxo-1-pyridyl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TPTU); O-benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU); (benzotriazol-1-yloxy)-tripyrrolidinophosphonium-hexafluorophosphate (PyBOP), O-(1H-6-chlorobenzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride/hydroxybenzotriazole or/1-hydroxy-7-azabenzotriazole (EDC/HOBT or EDC/HOAt) or HOAt alone, or with (1-chloro-2-methyl-propenyl)-dimethylamine. For review of some other possible coupling agents, see e.g. Klauser; Bodansky, Synthesis (1972), 453-463. The reaction mixture is preferably stirred at a temperature of between approximately −20 and 50° C., especially between 0° C. and 30° C., e.g. at room temperature.
In a compound of the formula I wherein R1 or especially R4 carries two vicinal amino groups, the two nitrogen atoms of the two amino groups can be bridged by a —CH═ group (thus forming, together with the two carbon atoms that bind the original amino groups and the bond between them, an 1H-imidazolo ring annelated to R1 or R4; for example, (vicinal diamino)-phenyl can be converted into benzoimidazolyl according to this method. The reaction preferably takes place by first reacting the compound of the formula I carrying the two vicinal amino groups with formic acid, e.g. in the presence of a coupling agent as mentioned in the preceding paragraph, such as EDC hydrochloride, a base, such as N,N-dimethylaminopyridine (DMAP) and preferably an appropriate solvent, such as methylene chloride, e.g. at temperatures in the range from −20 to 50° C., e.g. at about room temperature, thus converting one (especially a para-positioned) of the vicinal amino groups into a formylamino group. In a second step, the amino and formylamino group are then reacted to —N═C—N— by heating in the presence of an acid, especially acetic acid, e.g. at temperatures in the range from 50 to 110° C., for example at about 100° C.
Salts of a compound of formula I with a salt-forming group may be prepared in a manner known per se. Acid addition salts of compounds of formula I may thus be obtained by treatment with an acid or with a suitable anion exchange reagent. A salt with two acid molecules (for example a dihalogenide of a compound of formula I) may also be converted into a salt with one acid molecule per compound (for example a monohalogenide); this may be done by heating to a melt, or for example by heating as a solid under a high vacuum at elevated temperature, for example from 130 to 170° C., one molecule of the acid being expelled per molecule of a compound of formula I. Salts can usually be converted to free compounds, e.g. by treating with suitable basic compounds, for example with alkali metal carbonates, alkali metal hydrogencarbonates, or alkali metal hydroxides, typically potassium carbonate or sodium hydroxide.
Stereoisomeric mixtures, e.g. mixtures of diastereomers, can be separated into their corresponding isomers in a manner known per se by means of suitable separation methods. Diastereomeric mixtures for example may be separated into their individual diastereomers by means of fractionated crystallization, chromatography, solvent distribution, and similar procedures. This separation may take place either at the level of a starting compound or in a compound of formula I itself. Enantiomers may be separated through the formation of diastereomeric salts, for example by salt formation with an enantiomer-pure chiral acid, or by means of chromatography, for example by HPLC, using chromatographic substrates with chiral ligands.
It should be emphasized that reactions analogous to the conversions mentioned in this chapter may also take place at the level of appropriate intermediates (and are thus useful in the preparation of corresponding starting materials).
Starting Materials:
The starting materials of the formulae II, III and IV, as well as other starting materials mentioned herein, e.g. below, can be prepared according to or in analogy to methods that are known in the art, are known in the art and/or are commercially available. Novel starting materials, as well as processes for the preparation thereof, are likewise an embodiment of the present invention. In the preferred embodiments, such starting materials are used and the reaction chosen are selected so as to enable the preferred compounds to be obtained.
In the starting materials (including intermediates), which may also be used and/or obtained as salts where appropriate and expedient, the substituents are preferably as defined for a compound of the formula I.
The following examples illustrate the invention without limiting the scope thereof.
Temperatures are measured in degrees Celsius. Unless otherwise indicated, the reactions take place at rt.
The Rf values in TLC indicate the ratio of the distance moved by each substance to the distance moved by the eluent front. Rf values for TLC are measured on 5×10 cm TLC plates, silica gel F254, Merck, Darmstadt, Germany; the solvent systems are marked in the examples as follows:
If not indicated otherwise, the analytical HPLC conditions are as follows:
Abbreviations
Several aryl bromides and anilines as used according to Scheme 1-3 have been purchased from commercial sources where practicable. Otherwise, the aryl bromides and anilines/aminoheterocycles are being prepared according to the exemplified general procedures:
The compound shown on the left above, 2-(4-bromo-phenyl)-1-morpholin-4-yl-ethanone, is obtained by reaction of the corresponding acid chloride (A), (4-bromo-phenyl)-acetyl chloride, with morpholine and Et3N in DCM at rt. The product is obtained in high yield. The intermediate acid chloride (A) is obtained by reaction of (4-bromo-phenyl)-acetic acid (B) and oxalyl chloride (C) in DCM at rt and using DMF as reaction initiator. The intermediate (A) is obtained in good yield.
Alternatively, amide bond formation can be achieved by coupling of the appropriate carboxylic acid (B) and the appropriate amine in the presence of HATU and N-methyl morpholine in DMF at rt.
The following aryl bromides used in the examples below are synthesized analogously, using the appropriate starting materials:
The compound shown on the left above, 4-(4-Bromo-2-methyl-benzyl)-morpholine, is obtained by reaction of the corresponding benzyl bromide (A), 4-bromo-1-bromomethyl-2-methyl-benzene, with morpholine in DMF at rt. The product is obtained in high yield. The intermediate benzyl bromide (A) is obtained by reaction of (4-bromo-2-methyl-phenyl)-methanol (B) with PPh3 and CBr4 in DCM at rt. The intermediate (A) is obtained in high yield. The intermediate benzyl alcohol B is obtained by reduction of the corresponding carboxylic acid (C), 4-bromo-2-methyl-benzoic acid with LAH in THF. The intermediate (B) is obtained in good yield.
The following aryl bromides used in the examples below are synthesized analogously, using the appropriate corresponding starting materials:
The compound shown on the left above, (4-amino-phenyl)-morpholin-4-yl-methanone, is obtained by hydrogenation of the corresponding nitro-compound (A), morpholin-4-yl-(4-nitro-phenyl)-methanone, with Pd/C or Raney-Nickel and H2 or NH4CO2H in MeOH at rt. Alternatively, the nitro group reduction can be achieved with SnCl2 in EtOH at 90° C. The intermediate nitro compound (A) is obtained by the reaction of 4-nitro-benzoyl chloride (B) and morpholine and Et3N in DCM at rt. The intermediate (A) is obtained in good yield. When needed, analogs of (B) are obtained from the corresponding carboxylic acid derivatives heated to reflux for 2 hrs with thionylchloride or oxalylchloride in DCM or dichloroethane.
Alternatively, amide bond formation can be achieved by coupling of the appropriate carboxylic acid derivative (B) and the appropriate amine in the presence of TBTU and Et3N in THF at 0° C. to rt.
The following anilines used in the examples below are synthesized analogously, using the appropriate corresponding starting materials:
The compound shown on the left above, (4-amino-phenyl)-(4-methyl-piperazin-1-yl)-methanone, is obtained by treatment of intermediate (A), [4-(4-methyl-piperazine-1-carbonyl)-phenyl]-carbamic acid tert-butyl ester, with TFA in DCM at rt. The intermediate compound (A) is obtained by coupling of 4-tert-butoxycarbonylamino-benzoic acid (B) and N-methyl piperazine (C) in the presence of HATU and N-methyl morpholine in DCM at rt. The intermediate (A) is obtained in good yield.
The use of the Boc protecting group is not necessary for the preparation of certain aniline building blocks.
The following anilines used in the examples below are synthesized analogously, using the appropriate corresponding starting materials:
The compound shown on the left above, 3-(2-dimethylamino-ethoxy)-4-methoxy-phenylamine, is obtained by treatment of intermediate (A), [2-(2-methoxy-5-nitro-phenoxy)-ethyl]-dimethyl-amine, with Pd/C and NH4CO2H in MeOH/THF at rt. The intermediate compound (A) is obtained by alkylation of 2-methoxy-5-nitro-phenol (B) with (2-chloro-ethyl)-dimethyl-amine (C) in the presence of NaH in DMF at 150° C.
The following anilines used in the examples below are synthesized analogously, using the appropriate corresponding starting materials:
The compound shown on the left above, 4-(cis-3,5-dimethyl-piperazin-1-ylmethyl)-phenylamine, is obtained by treatment of intermediate (A), cis-3,5-dimethyl-1-(4-nitro-benzyl)-piperazine, with SnCl2 hydrate in MeOH at rt. Alternatively, the nitro group reduction can be achieved with Pd/C or Raney-Nickel and H2 or NH4CO2H in MeOH at rt. The intermediate compound (A) is obtained by reaction of cis-2,6-dimethyl-piperazine (B) with 1-bromomethyl-4-nitro-benzene (C) in the presence of Et3N or DIPEA in DMF at rt.
Where needed, the (substituted) benzyl bromide (C) is synthesized from the corresponding carboxylic acid (D) via reduction to the alcohol and conversion to the halogen compound.
The following anilines used in the examples below are synthesized analogously, using the appropriate corresponding starting materials:
Alternatively, (A) is obtained by the reaction of 2-bromomethyl-1-methyl-4-nitro-benzene (C) with sodium azide in ethanol/water 1:2 at rt overnight, followed by triazole formation with dimethyl-prop-2-ynyl-amine in tert-butanol/water 1:1 in presence of cupper (I) sulfate (0.15 eq.) and L-(+)-sodium ascorbate (0.3 eq.).
Alternatively, (B) is obtained from the reaction of 3-oxo-piperazine-1-carboxylic acid tert-butyl ester activated with sodium hydride with (2-bromo-ethyl)-dimethyl-amine in DMF. The resulting material is then Boc-deprotected in dioxane with HCl to give (B) as HCl salt.
The compound shown on the left above, 6-(cis-3,5-dimethyl-piperazin-1-yl)-pyridin-3-ylamine, is obtained by treatment of intermediate (A), cis-3,5-dimethyl-1-(5-nitro-pyridin-2-yl)-piperazine, with H2 (or ammonium formate) and Pd/C (or Raney-Nickel) in MeOH (and THF) at rt. The intermediate compound (A) is obtained by reaction of cis-2,6-dimethyl-piperazine (B) with 2-chloro-5-nitro-pyridine (C) in the presence of Et3N in THF at rt to 70° C.
Alternatively, in case (B) is an alcohol, 2-bromo-5-nitro-pyridine, or 2-chloro-4-nitro-pyridine-1-oxide for some of the meta-substituted derivatives, is used as (C) in presence of KOtBu or Cs2CO3 or NaH in DMA or THF at rt to 80° C.
The following anilines used in the examples below are synthesized analogously, using the appropriate corresponding starting materials:
The compound shown on the left above, 4-(4-ethyl-piperazin-1-yl)-3-methyl-phenylamine, is obtained by treatment of intermediate (A), 1-ethyl-4-(2-methyl-4-nitro-phenyl)-piperazine, with H2 (or ammonium formate) and Pd/C in MeOH (and THF) at rt. The intermediate compound (A) is obtained by reaction of N-ethyl-piperazine (B) with 1-fluoro-2-methyl-4-nitro-benzene (C) in dimethylacetamide (DMA) at 110° C. for 20 hours.
The compound shown on the left above, 1-(2-methoxy-ethyl)-1H-pyrazol-4-ylamine, is obtained by treatment of intermediate (A), 1-(2-methoxy-ethyl)-4-nitro-1H-pyrazole, with ammonium formate and Pd/C in MeOH at rt. The intermediate compound (A) is obtained by alkylation of 4-nitro-1H-pyrazole (B) with 1-chloro-2-methoxy-ethane (C) in the presence of NaH in DMF at 95° C.
The following aminopyrazole used in the examples below is synthesized analogously, using the appropriate corresponding starting materials:
The compound shown on the left above, 2-(4-methyl-piperazin-1-ylmethyl)-pyridin-4-ylamine), is obtained by treatment of intermediate (A), N-[2-(4-methyl-piperazin-1-ylmethyl)-pyridin-4-yl]-acetamide, with KOH in EtOH/water 2:1 at 100° C. for 4 h. The intermediate compound (A) is obtained by reaction of N-methyl-piperazine (C) with methanesulfonic acid 4-acetylamino-pyridin-2-ylmethyl ester (B) in DCM at rt in the presence of DIPEA. The intermediate compound (B) is obtained from the reaction of N-(2-hydroxymethyl-pyridin-4-yl)-acetamide (D) with methanesulfonyl chloride in the presence of Et3N in DCM (or with SOCl2 in 1,2-dichloroethane with a small amount of DMF for the chloride analog of (B)). Intermediate (D) is obtained from the treatment of (4-chloro-pyridin-2-yl)-methanol with acetamide in presence of Pd(OAc)2, 4,5-bis-diphenylphosphanyl-9,9-dimethyl-9H-xanthene and cesium carbonate in dioxane and DMF at 130° C. in the microwave oven for 1 h.
The following anilines used in the examples below are synthesized analogously, using the appropriate corresponding starting materials:
The compound shown on the left above, (S)-3-(4-amino-phenyl)-morpholine-4-carboxylic acid tert-butyl ester, is obtained from the hydrogenation of intermediate (A) with hydrazine hydrate in ethanol in presence of Raney-Nickel under argon atmosphere at rt, followed by separation of the NO2-regioisomers. To obtain the intermediate compound (A) (mixture of regioisomers), a solution of (S)-3-phenyl-morpholine (B) in DCM is slowly dropped into fuming nitric acid in DCM at −45° C. After 1 h, the reaction is quenched slowly with an aq. sol. of NaOH followed by a DCM/aq. bicarbonate work-up. The resulting material is purified on silica column chromatography and then Boc protected with Boc-anhydride in DCM at rt.
The intermediate compound (B) is obtained from the reduction of (S)-5-phenyl-morpholin-3-one (C) with LAH in THF at rt. The morpholine derivative compound (C) is obtained from the ring closing of 2-chloro-N-((S)-2-hydroxy-1-phenyl-ethyl)-acetamide (D) in presence of sodium hydride in THF/toluene 1:1 at 0° C. to rt. The intermediate compound (D) results from the slow addition of chloro-acetyl chloride (F) dissolved in DCM to (S)-2-amino-2-phenyl-ethanol (E) in THF in presence of Et3N at 0° C. to rt.
The following anilines used in the examples below are synthesized analogously, using the appropriate corresponding starting materials:
The compound is prepared according to Scheme 1.
2,4-Dichloro-5-bromopyrimidine (200 g, 0.88 mol) is added slowly to NH3 (1000 ml, 7 M in MeOH) while the reaction mixture is kept below 10° C. The reaction mixture is stirred at rt for 2 h, heated to 60° C. for 2 h, and then cooled again to rt and stirred for 55 h. It is concentrated under reduced pressure, and the residue is suspended in H2O (500 ml). The aqueous layer is extracted with EtOAc (3×) and the combined organic layers are dried over MgSO4, filtered, and concentrated under reduced pressure to yield the title compound as a white solid.
To a solution of ethoxyethyne (120 g, 0.69 mol, 40% in hexane) in toluene (1500 mL) is added slowly tributyltin hydride (190 g, 0.65 mol) and AlBN (4.6 g, 0.028 mol). The reaction mixture is heated to 100° C. for 16 h. The reaction mixture is concentrated under reduced pressure and dried under HV. The brown residual oil (80% purity of title compound) is used for next step without purification.
5-Bromo-2-chloro-pyrimidin-4-ylamine (88.0 g, 0.43 mol), 1-ethoxy-propene (220 g, 0.49 mol), Pd(PPh3)2Cl2 (35.0 g, 0.05 mol) and Et4NCl (67.0 g, 0.40 mol) are suspended in solvent (750 mL, DME/toluene/H2O/EtOH 10:1:3:6) under nitrogen. The reaction mixture is heated to reflux for 24 h, cooled to rt, and then diluted with water. The aqueous layer is extracted with EtOAc (3×). The combined organic layers are dried over MgSO4, filtered, and concentrated under reduced pressure. The residue is purified by column chromatography (SiO2, gradient elution, EtOAc/petroleumether 1:10>1:4) to yield the title compound as a yellow solid.
To a solution of 2-Chloro-5-(2-ethoxy-vinyl)-pyrimidin-4-ylamine (38.0 g, 0.19 mol) in ETOH (1000 ml) is added concentrated aqueous HCl (37%, 100 g, 1.00 mol) at rt. The reaction mixture is heated to reflux for 3 h and then evaporated to dryness under reduced pressure. Aqueous sodium carbonate solution (5%, 500 mL) is added to the residue, and the mixture is extracted with EtOAc (3×). The combined organic layers are dried over MgSO4, filtered, and concentrated under reduced pressure. The residue is recrystallized from hexane/ether (4/1, 250 ml) to give the title compound as an off-white solid.
In a seal tube, 2-chloro-7H-pyrrolo[2,3-d]pyrimidine (600 mg, 3.56 mmol), 4-bromophenylmethylsulfone (700 mg, 4.10 mmol), CuI (239 mg, 1.23 mmol), and K3PO4 (2.67 g, 12.3 mmol) are suspended in 1,4-dioxane (30 mL). Then, trans-1,2-diaminocyclohexane (149 μL, 1.23 mmol) is added at rt. The reaction vial is flushed with Ar and the mixture is heated to 110° C. for 3 h. After cooling to rt, the reaction mixture is concentrated under reduce pressure. The residue is suspended in EtOAc and washed with saturated aqueous NaCl solution (3×). The organic layer is dried over MgSO4, filtered, and concentrated under reduced pressure. The solid residue is triturated with small amounts of EtOAc to yield the title compound as a brown solid.
To a suspension of 2-chloro-7-(4-methanesulfonyl-phenyl)-7H-pyrrolo[2,3-d]pyrimidine (30.0 mg, 0.093 mmol) in BuOH (1.5 mL) is added 3,4-diethoxyaniline (41.0 mg, 0.220 mmol) and concentrated aqueous HCl (37%, 23.0 μL, 0.232 mmol). The reaction mixture is heated to 140° C. for 24 h, cooled to rt, and concentrated under reduce pressure. The residue is purified by reverse phase prep-HPLC (Waters) to afford the title compound (1) as a yellow solid. HPLC: tR=1.61 min (Method A); MS-ES: (M+H)+=453.
The compound is prepared according to Scheme 2.
In a sealed tube, 2-chloro-7H-pyrrolo[2,3-d]pyrimidine (200 mg, 1.24 mmol), 4-(4-bromo-2,6-difluoro-benzyl)-morpholine (418 mg, 1.36 mmol), CuI (72.1 mg, 0.371 mmol), and K3PO4 (804 mg, 3.71 mmol) are suspended in 1,4-dioxane (8 mL). Then, trans-1,2-diaminocyclohexane (45.0 μL, 0.371 mmol) is added at rt. The reaction vial is flushed with Ar and the mixture is heated to 110° C. for 5 h. After cooling to rt, the reaction mixture is diluted with EtOAc and the organic layer is washed with saturated aqueous Na2CO3 solution (2×). The organic layer is dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue is purified by column chromatography (SiO2, gradient elution, hexane/EtOAc 100:0→30:70) to yield the title compound as a white solid.
In a sealed tube, 2-chloro-7-(3,5-difluoro-4-morpholin-4-ylmethyl-phenyl)-7H-pyrrolo[2,3-d]pyrimidine (50.0 mg, 0.130 mmol), (4-Amino-phenyl)-(4-methyl-piperazin-1-yl)-methanone (42.1 mg, 0.182 mmol), KOtBu (21.1 mg, 0.182 mmol) and SK-CC02-A (12.5 mg, 0.020 mmol, Pd catalyst 2-(Dimethylaminomethyl)-ferrocen-1-yl-palladium(II)-chlorid Dinorbornylphosphin Complex, Fluka No. 44696) are suspended in THF (2 ml) under Ar. The reaction mixture is stirred at 80° C. for 1.5 h, cooled to rt, and then filtered through a Celite plug. The filtrate is concentrated under reduce pressure. The residue is purified by reverse phase prep-HPLC (Waters) to afford the title compound (2) as a white solid. HPLC: tR=0.89 min (Method A); MS-ES: (M+H)+=548.
The compound is prepared according to Scheme 3.
To a suspension of 2-chloro-7H-pyrrolo[2,3-d]pyrimidine (700 mg, 4.33 mmol) in BuOH (10 mL) is added 4-isopropoxyaniline (1.31 g, 8.66 mmol) and concentrated aqueous HCl (37%, 1.28 mL, 13.0 mmol). The reaction mixture is heated to 140° C. for 24 h, cooled to rt, and concentrated under reduce pressure. The residue is dissolved in EtOAc and the organic layer is washed with saturated aqueous Na2CO3 solution (2×). The organic layer is dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue solid residue is triturated with hexane to yield the title compound as an off-white solid.
In a sealed tube, (4-Isopropoxy-phenyl)-(7H-pyrrolo[2,3-d]pyrimidin-2-yl)-amine (350 mg, 1.17 mmol), 4-bromophenylmethylsulfone (552 mg, 2.35 mmol), CuI (68.4 mg, 0.352 mmol), and K3PO4 (763 mg, 3.52 mmol) are suspended in 1,4-dioxane (10 mL). Then, trans-1,2-diaminocyclohexane (42.7 μL, 0.352 mmol) is added at rt. The reaction vial is flushed with Ar and the mixture is heated to 110° C. for 6 h. After cooling to rt, the reaction mixture is diluted with EtOAc and the organic layer is washed with saturated aqueous NaCl solution (3×). The organic layer is dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue is purified by reverse phase prep-HPLC (Waters) to afford the title compound (3) as an off-white solid. HPLC: tR=1.67 min (Method A); MS-ES: (M+H)+=423.
The compound is prepared analogous to Example 1. HPLC: tR=1.48 min (Method A); MS-ES: (M+H)+=455.
The compound is prepared analogous to Example 3. HPLC: tR=1.37 min (Method A); MS-ES: (M+H)+=425.
The compound is prepared analogous to Example 3. HPLC: tR=1.73 min (Method A); MS-ES: (M+H)+=482.
The compound is prepared analogous to Example 3. HPLC: tR=1.71 min (Method A); MS-ES: (M+H)+=446.
The compound is prepared analogous to Example 3. HPLC: tR=1.83 min (Method A); MS-ES: (M+H)+=512.
The compound is prepared analogous to Example 3. HPLC: tR=1.81 min (Method A); MS-ES: (M+H)+=476.
The compound is prepared analogous to Example 3. HPLC: tR=1.69 min (Method A); MS-ES: (M+H)+=484.
The compound is prepared analogous to Example 1. HPLC: tR=1.32 min (Method A); MS-ES: (M+H)+=439.
The compound is prepared analogous to Example 1. HPLC: tR=1.06 min (Method A); MS-ES: (M+H)+=505.
The compound is prepared analogous to Example 1. HPLC: tR=1.44 min (Method A); MS-ES: (M+H)+=548.
The compound is prepared analogous to Example 1. HPLC: tR=2.30 min (Method A); MS-ES: (M+H)+=495.
The compound is prepared analogous to Example 1. HPLC: tR=1.42 min (Method A); MS-ES: (M+H)+=409.
The compound is prepared analogous to Example 1. HPLC: tR=1.08 min (Method A); MS-ES: (M+H)+=462.
The compound is prepared analogous to Example 1. HPLC: tR=1.09 min (Method A); MS-ES: (M+H)+=463.
The compound is prepared analogous to Example 1. HPLC: tR=0.98 min (Method A); MS-ES: (M+H)+=463.
The compound is prepared analogous to Example 1. HPLC: tR=1.46 min (Method A); MS-ES: (M+H)+=365.
The compound is prepared analogous to Example 1. HPLC: tR=1.41 min (Method A); MS-ES: (M+H)+=395.
The compound is prepared analogous to Example 1. HPLC: tR=1.07 min (Method A); MS-ES: (M+H)+=408.
The compound is prepared analogous to Example 1. MS-ES: (M+H)+=425.
The compound is prepared analogous to Example 1. HPLC: tR=1.62 min (Method A); MS-ES: (M+H)+=425.
The compound is prepared analogous to Example 1. HPLC: tR=1.22 min (Method A); MS-ES: (M+H)+=411.
The compound is prepared analogous to Example 1. HPLC: tR=1.07 min (Method A); MS-ES: (M+H)+=493.
The compound is prepared analogous to Example 1. HPLC: tR=1.09 min (Method A); MS-ES: (M+H)+=482.
The compound is prepared analogous to Example 1. HPLC: tR=1.10 min (Method A); MS-ES: (M+H)+=496.
The compound is prepared analogous to Example 2. HPLC: tR=1.02 min (Method A); MS-ES: (M+H)+=491.
The compound is prepared analogous to Example 1. HPLC: tR=1.11 min (Method A); MS-ES: (M+H)+=524.
The compound is prepared analogous to Example 2. HPLC: tR=1.33 min (Method A); MS-ES: (M+H)+=478.
The compound is prepared analogous to Example 1. HPLC: tR=1.13 min (Method A); MS-ES: (M+H)+=508.
The compound is prepared analogous to Example 1. HPLC: tR=1.05 min (Method A); MS-ES: (M+H)+=507.
The compound is prepared analogous to Example 1. HPLC: tR=1.09 min (Method A); MS-ES: (M+H)+=480.
The compound is prepared analogous to Example 1. HPLC: tR=1.01 min (Method A); MS-ES: (M+H)+=494.
The compound is prepared analogous to Example 1. HPLC: tR=1.13 min (Method A); MS-ES: (M+H)+=494.
The compound is prepared analogous to Example 1. HPLC: tR=1.20 min (Method A); MS-ES: (M+H)+=480.
The compound is prepared analogous to Example 1. HPLC: tR=0.98 min (Method A); MS-ES: (M+H)+=449.
The compound is prepared analogous to Example 2. HPLC: tR=0.98 min (Method A); MS-ES: (M+H)+=477.
The compound is prepared analogous to Example 2. HPLC: tR=1.02 min (Method A); MS-ES: (M+H)+=491.
The compound is prepared analogous to Example 2. HPLC: tR=1.10 min (Method A); MS-ES: (M+H)+=479.
The compound is prepared analogous to Example 2. HPLC: tR=1.16 min (Method A); MS-ES: (M+H)+=462.
The compound is prepared analogous to Example 2. HPLC: tR=1.15 min (Method A); MS-ES: (M+H)+=462.
The compound is prepared analogous to Example 2. HPLC: tR=1.06 min (Method A); MS-ES: (M+H)+=445.
The compound is prepared analogous to Example 2. HPLC: tR=1.08 min (Method A); MS-ES: (M+H)+=464.
The compound is prepared analogous to Example 2. HPLC: tR=1.41 min (Method A); MS-ES: (M+H)+=563.
The compound is prepared analogous to Example 2. HPLC: tR=1.92 min (Method A); MS-ES: (M+H)+=534.
The compound is prepared analogous to Example 2. HPLC: tR=1.08 min (Method A); MS-ES: (M+H)+=445.
The compound is prepared analogous to Example 2. HPLC: tR=1.28 min (Method A); MS-ES: (M+H)+=555.
The compound is prepared analogous to Example 2. HPLC: tR=1.32 min (Method A); MS-ES: (M+H)+=478.
The compound is prepared analogous to Example 2. HPLC: tR=1.04 min (Method A); MS-ES: (M+H)+=479.
The compound is prepared analogous to Example 2. HPLC: tR=1.17 min (Method A); MS-ES: (M+H)+=452.
The compound is prepared by treatment of Example 45 with TFA in CH2Cl2 at rt. HPLC: tR=1.08 min (Method A); MS-ES: (M+H)+=464.
The compound is prepared by treatment of Example 46 with TFA in CH2Cl2 at rt. HPLC: tR=1.09 min (Method A); MS-ES: (M+H)+=434.
The compound is prepared analogous to Example 2. HPLC: tR=1.46 min (Method A); MS-ES: (M+H)+=409.
The compound is prepared analogous to Example 1. HPLC: tR=1.52 min (Method A); MS-ES: (M+H)+=383.
The compound is prepared analogous to Example 3. HPLC: tR=1.06 min (Method A); MS-ES: (M+H)+=431.
The compound is prepared analogous to Example 3. HPLC: tR=1.56 min (Method A); MS-ES: (M+H)+=402.
The compound is prepared analogous to Example 3. HPLC: tR=1.11 min (Method A); MS-ES: (M+H)+=449.
The compound is prepared analogous to Example 3. HPLC: tR=1.54 min (Method A); MS-ES: (M+H)+=436.
The compound is prepared analogous to Example 3. HPLC: tR=1.71 min (Method A); MS-ES: (M+H)+=347.
The compound is prepared analogous to Example 3. HPLC: tR=1.42 min (Method A); MS-ES: (M+H)+=418.
The compound is prepared analogous to Example 3. HPLC: tR=1.13 min (Method A); MS-ES: (M+H)+=461.
The compound is prepared analogous to Example 3. HPLC: tR=1.23 min (Method A); MS-ES: (M+H)+=459.
The compound is prepared analogous to Example 2. HPLC: tR=1.11 min (Method A); MS-ES: (M+H)+=512.
The compound is prepared analogous to Example 2. HPLC: tR=1.04 min (Method A); MS-ES: (M+H)+=493.
The compound is prepared analogous to Example 2. HPLC: tR=1.02 min (Method A); MS-ES: (M+H)+=519.
The compound is prepared analogous to Example 2. HPLC: tR=1.10 min (Method A); MS-ES: (M+H)+=521.
The compound is prepared analogous to Example 2. HPLC: tR=1.18 min (Method A); MS-ES: (M+H)+=519.
The compound is prepared analogous to Example 2. HPLC: tR=1.80 min (Method A); MS-ES: (M+H)+=577.
The compound is prepared analogous to Example 2. HPLC: tR=1.03 min (Method A); MS-ES: (M+H)+=505.
The compound is prepared analogous to Example 2. HPLC: tR=1.13 min (Method A); MS-ES: (M+H)+=509.
The compound is prepared analogous to Example 2. HPLC: tR=1.23 min (Method A); MS-ES: (M+H)+=559.
The compound is prepared analogous to Example 2. HPLC: tR=1.02 min (Method A); MS-ES: (M+H)+=505.
The compound is prepared analogous to Example 2. HPLC: tR=1.31 min (Method A); MS-ES: (M+H)+=492.
The compound is prepared analogous to Example 2. HPLC: tR=1.42 min (Method A); MS-ES: (M+H)+=476.
The compound is prepared analogous to Example 2. HPLC: tR=1.06 min (Method A); MS-ES: (M+H)+=509.
The compound is prepared analogous to Example 2. HPLC: tR=1.38 min (Method A); MS-ES: (M+H)+=496.
The compound is prepared analogous to Example 2. HPLC: tR=1.14 min (Method A); MS-ES: (M+H)+=537.
The compound is prepared analogous to Example 2. HPLC: tR=1.61 min (Method A); MS-ES: (M+H)+=407.
The compound is prepared analogous to Example 2. HPLC: tR=2.01 min (Method A); MS-ES: (M+H)+=451.
The compound is prepared analogous to Example 2. HPLC: tR=1.67 min (Method A); MS-ES: (M+H)+=397.
The compound is prepared analogous to Example 3. HPLC: tR=1.76 min (Method A); MS-ES: (M+H)+=494.
The compound is prepared analogous to Example 3. HPLC: tR=1.79 min (Method A); MS-ES: (M+H)+=500.
The compound is prepared analogous to Example 3. HPLC: tR=1.69 min (Method A); MS-ES: (M+H)+=498.
The compound is prepared analogous to Example 3. HPLC: tR=1.23 min (Method A); MS-ES: (M+H)+=489.
The compound is prepared analogous to Example 3. HPLC: tR=1.83 min (Method A); MS-ES: (M+H)+=387.
The compound is prepared by treatment of Example 69 with TFA in CH2Cl2 at rt. MS-ES: (M+H)+=477.
The compound is prepared analogous to Example 3. HPLC: tR=1.24 min (Method A); MS-ES: (M+H)+=463.
The compound is prepared analogous to Example 3. HPLC: tR=1.78 min (Method A); MS-ES: (M+H)+=468.
The compound is prepared analogous to Example 3. MS-ES: (M+H)+=466.
The compound is prepared analogous to Example 3. MS-ES: (M+H)+=446.
The compound is prepared analogous to Example 3. MS-ES: (M+H)+=450.
The compound is prepared analogous to Example 3. HPLC: tR=1.02 min (Method A); MS-ES: (M+H)+=477.
The compound is prepared analogous to Example 3. HPLC: tR=1.15 min (Method A); MS-ES: (M+H)+=525.
The compound is prepared analogous to Example 3. HPLC: tR=1.77 min (Method A); MS-ES: (M+H)+=390.
The compound is prepared analogous to Example 3. HPLC: tR=1.90 min (Method A); MS-ES: (M+H)+=370.
The compound is prepared analogous to Example 3. HPLC: tR=1.27 min (Method A); MS-ES: (M+H)+=462.
The compound is prepared analogous to Example 3. HPLC: tR=1.64 min (Method A); MS-ES: (M+H)+=476.
The compound is prepared analogous to Example 3. HPLC: tR=1.69 min (Method A); MS-ES: (M+H)+=492.
The compound is prepared analogous to Example 3. HPLC: tR=1.61 min (Method A); MS-ES: (M+H)+=472.
The compound is prepared analogous to Example 3. HPLC: tR=1.27 min (Method A); MS-ES: (M+H)+=485.
The compound is prepared analogous to Example 3. HPLC: tR=1.60 min (Method A); MS-ES: (M+H)+=472.
The compound is prepared analogous to Example 3. HPLC: tR=1.36 min (Method A); MS-ES: (M+H)+=511.
The compound is prepared analogous to Example 3. HPLC: tR=1.13 min (Method A); MS-ES: (M+H)+=443.
The compound is prepared analogous to Example 3. HPLC: tR=1.72 min (Method A); MS-ES: (M+H)+=428.
The compound is prepared analogous to Example 3. HPLC: tR=1.25 min (Method A); MS-ES: (M+H)+=489.
The compound is prepared analogous to Example 3. HPLC: tR=1.27 min (Method A); MS-ES: (M+H)+=428.
The compound is prepared analogous to Example 3. HPLC: tR=1.14 min (Method A); MS-ES: (M+H)+=475.
The compound is prepared analogous to Example 3. HPLC: tR=1.23 min (Method A); MS-ES: (M+H)+=444.
The compound is prepared analogous to Example 3. HPLC: tR=1.33 min (Method A); MS-ES: (M+H)+=478.
The compound is prepared analogous to Example 3. HPLC: tR=1.28 min (Method A); MS-ES: (M+H)+=458.
The compound is prepared analogous to Example 3. HPLC: tR=1.55 min (Method A); MS-ES: (M+H)+=510.
The compound is prepared analogous to Example 3. HPLC: tR=1.73 min (Method A); MS-ES: (M+H)+=526.
The compound is prepared analogous to Example 3. HPLC: tR=1.63 min (Method A); MS-ES: (M+H)+=389.
The compound is prepared analogous to Example 3. HPLC: tR=1.27 min (Method A); MS-ES: (M+H)+=458.
The compound is prepared analogous to Example 3. HPLC: tR=2.03 min (Method A); MS-ES: (M+H)+=484.
The compound is prepared analogous to Example 3. HPLC: tR=1.47 min (Method A); MS-ES: (M+H)+=501.
The compound is prepared analogous to Example 3. MS-ES: (M+H)+=452.
The compound is prepared analogous to Example 3. HPLC: tR=1.33 min (Method A); MS-ES: (M+H)+=434.
The compound is prepared analogous to Example 3. HPLC: tR=1.38 min (Method A); MS-ES: (M+H)+=479.
The compound is prepared analogous to Example 3. HPLC: tR=1.73 min (Method A); MS-ES: (M+H)+=500.
The compound is prepared analogous to Example 3. HPLC: tR=1.33 min (Method A); MS-ES: (M+H)+=485.
The compound is prepared analogous to Example 3. HPLC: tR=1.18 min (Method A); MS-ES: (M+H)+=418.
The compound is prepared analogous to Example 3. HPLC: tR=1.24 min (Method A); MS-ES: (M+H)+=463.
The compound is prepared analogous to Example 3. HPLC: tR=1.53 min (Method A); MS-ES: (M+H)+=484.
The compound is prepared analogous to Example 3. HPLC: tR=1.32 min (Method A); MS-ES: (M+H)+=480.
The compound is prepared analogous to Example 3. HPLC: tR=1.26 min (Method A); MS-ES: (M+H)+=436.
The compound is prepared analogous to Example 3. HPLC: tR=1.48 min (Method A); MS-ES: (M+H)+=470.
The compound is prepared analogous to Example 3. HPLC: tR=1.96 min (Method A); MS-ES: (M+H)+=518.
The compound is prepared analogous to Example 3. HPLC: tR=1.51 min (Method A); MS-ES: (M+H)+=497.
The compound is prepared analogous to Example 3. HPLC: tR=1.39 min (Method A); MS-ES: (M+H)+=454.
The compound is prepared analogous to Example 3. HPLC: tR=1.35 min (Method A); MS-ES: (M+H)+=481.
The compound is prepared analogous to Example 3. HPLC: tR=1.26 min (Method A); MS-ES: (M+H)+=495.
The compound is prepared analogous to Example 3. HPLC: tR=1.43 min (Method A); MS-ES: (M+H)+=495.
The compound is prepared analogous to Example 2. HPLC: tR=1.15 min (Method A); MS-ES: (M+H)+=466.
The compound is prepared analogous to Example 2. HPLC: tR=1.38 min (Method A); MS-ES: (M+H)+=447.
The compound is prepared analogous to Example 3. HPLC: tR=1.71 min (Method A); MS-ES: (M+H)+=502.
The compound is prepared analogous to Example 2. HPLC: tR=1.08 min (Method A); MS-ES: (M+H)+=535.
The compound is prepared analogous to Example 3. HPLC: tR=7.96 min. (Method B); MS-ES: (M+H)+=377; TLC***: Rf=0.22
The compound is prepared analogous to Example 3. HPLC: tR=8.23 min. (Method B); MS-ES: (M+H)+=393; TLC***: Rf=0.42
The compound is prepared analogous to Example 3. HPLC: tR=8.36 min. (Method B); MS-ES: (M+H)+=428; TLCt: Rf=0.14
The compound is prepared analogous to Example 3. HPLC: tR=8.27 min. (Method B); MS-ES: (M+H)+=462; TLC‡: Rf=0.23
The compound is prepared analogous to Example 3. HPLC: tR=8.04 min. (Method B); MS-ES: (M+H)+=414; TLC‡: Rf=0.11
The compound is prepared analogous to Example 3. HPLC: tR=9.22 min. (Method B); MS-ES: (M+H)+=482; TLC‡: Rf=0.12
The compound is prepared analogous to Example 3. HPLC: tR=8.15 min. (Method B); MS-ES: (M+H)+=495; TLC*: Rf=0.29
The compound is prepared analogous to Example 3. HPLC: tR=7.28 min. (Method B); MS-ES: (M+H)+=400; TLC‡: Rf=0.17
The compound is prepared analogous to Example 3. HPLC: tR=7.47 min. (Method B); MS-ES: (M+H)+=474
The compound is prepared analogous to Example 3. HPLC: tR=7.50 min. (Method B); MS-ES: (M+H)+=455; TLC*: Rf=0.36
The compound is prepared analogous to Example 3. HPLC: tR=7.25 min. (Method B); MS-ES: (M+H)+=441; TLC*: Rf=0.37
The compound is prepared analogous to Example 3. HPLC: tR=7.21 min. (Method B); MS-ES: (M+H)+=475; TLC*: Rf=0.36
The compound is prepared analogous to Example 3. HPLC: tR=8.40 min. (Method B); MS-ES: (M+H)+=466; TLC‡: Rf=0.16
The compound is prepared analogous to Example 3. HPLC: tR=8.76 min. (Method B); MS-ES: (M+H)+=446; TLC‡: Rf=0.26
The compound is prepared analogous to Example 2. HPLC: tR=7.03 min. (Method B); MS-ES: (M+H)+=547; TLC**: Rf=0.29
The compound is prepared analogous to Example 3. HPLC: tR=7.26 min. (Method B); MS-ES: (M+H)+=479; TLC**: Rf=0.23
The compound is prepared analogous to Example 3. HPLC: tR=7.45 min. (Method B); MS-ES: (M+H)+=459; TLC**: Rf=0.31
The compound is prepared analogous to Example 3. HPLC: tR=8.51 min. (Method B); MS-ES: (M+H)+=508; TLC**: Rf=0.67
The compound is prepared analogous to Example 2. HPLC: tR=7.03 min. (Method B); MS-ES: (M+H)+=517; TLC*: Rf=0.29
The compound is prepared analogous to Example 3. HPLC: tR=6.71 min. (Method B); MS-ES: (M+H)+=548; TLC*: Rf=0.57
The compound is prepared analogous to Example 3. HPLC: tR=8.20 min. (Method B); MS-ES: (M+H)+=495; TLC**: Rf=0.11
The compound is prepared analogous to Example 3. HPLC: tR=7.80 min. (Method B); MS-ES: (M+H)+=414; TLC**: Rf=0.21
The compound is prepared analogous to Example 3. But during the synthesis of the pyrrolopyrimidine core, tributyl-(1-propynyl)-tin was used instead of tributyl-(2-ethoxy-vinyl)-stannane as described in Example 1 to obtain 2-chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidine. HPLC: tR=7.76 min. (Method B); MS-ES: (M+H)+=469; TLC**: Rf=0.31
The compound is prepared analogous to Example 3. But during the synthesis of the pyrrolopyrimidine core, tributyl-(1-propynyl)-tin was used instead of tributyl-(2-ethoxy-vinyl)-stannane as described in Example 1 to obtain 2-chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidine. HPLC: tR=7.31 min. (Method B); MS-ES: (M+H)+=489; TLC***: Rf=0.20
The compound is prepared analogous to Example 3. But during the synthesis of the pyrrolopyrimidine core, tributyl-(1-propynyl)-tin was used instead of tributyl-(2-ethoxy-vinyl)-stannane as described in Example 1 to obtain 2-chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidine. HPLC: tR=7.58 min. (Method B); MS-ES: (M+H)+=446; TLC***: Rf=0.40
The compound is prepared analogous to Example 3. But during the synthesis of the pyrrolopyrimidine core, tributyl-(1-propynyl)-tin was used instead of tributyl-(2-ethoxy-vinyl)-stannane as described in Example 1 to obtain 2-chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidine. HPLC: tR=7.23 min. (Method B); MS-ES: (M+H)+=466; TLC***: Rf=0.32
The compound is prepared analogous to Example 3. HPLC: tR=7.53 min. (Method B); MS-ES: (M+H)+=529; TLC*: Rf=0.43
The compound is prepared analogous to Example 3. HPLC: tR=8.63 min. (Method B); MS-ES: (M+H)+=442; TLC**: Rf=0.30
The compound is prepared analogous to Example 3. But during the synthesis of the pyrrolopyrimidine core, tributyl-(1-propynyl)-tin was used instead of tributyl-(2-ethoxy-vinyl)-stannane as described in Example 1 to obtain 2-chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidine. HPLC: tR=7.52 min. (Method B); MS-ES: (M+H)+=529; TLC*: Rf=0.50
The compound is prepared analogous to Example 3. But during the synthesis of the pyrrolopyrimidine core, tributyl-(1-propynyl)-tin was used instead of tributyl-(2-ethoxy-vinyl)-stannane as described in Example 1 to obtain 2-chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidine. HPLC: tR=7.49 min. (Method B); MS-ES: (M+H)+=506; TLC***: Rf=0.24
The compound is prepared analogous to Example 3. HPLC: tR=8.54 min. (Method B); MS-ES: (M+H)+=428; TLC*: Rf=0.66
The compound is prepared analogous to Example 3. HPLC: tR=7.36 min. (Method B); MS-ES: (M+H)+=441; TLC**: Rf=0.18
The compound is prepared analogous to Example 2. HPLC: tR=0.93 min (Method A); MS-ES: (M+H)+=562.
The compound is prepared analogous to Example 2. HPLC: tR=0.93 min (Method A); MS-ES: (M+H)+=576.
The compound is prepared analogous to Example 2. HPLC: tR=0.93 min (Method A); MS-ES: (M+H)+=534.
The compound is prepared analogous to Example 2. HPLC: tR=0.93 min (Method A); MS-ES: (M+H)+=520.
The compound is prepared analogous to Example 2. HPLC: tR=1.13 min (Method A); MS-ES: (M+H)+=533.
The compound is prepared analogous to Example 2. HPLC: tR=1.19 min (Method A); MS-ES: (M+H)+=464.
The compound is prepared analogous to Example 2. HPLC: tR=1.28 min (Method A); MS-ES: (M+H)+=508.
The compound is prepared analogous to Example 2. HPLC: tR=1.13 min (Method A); MS-ES: (M+H)+=519.
The compound is prepared analogous to Example 2. HPLC: tR=1.21 min (Method A); MS-ES: (M+H)+=450.
The compound is prepared analogous to Example 2. HPLC: tR=1.53 min (Method A); MS-ES: (M+H)+=470.
The compound is prepared analogous to Example 2. HPLC: tR=1.20 min (Method A); MS-ES: (M+H)+=471.
The compound is prepared analogous to Example 2. HPLC: tR=1.17 min (Method A); MS-ES: (M+H)+=471.
The compound is prepared analogous to Example 2. HPLC: tR=1.21 min (Method A); MS-ES: (M+H)+=471.
The compound is prepared analogous to Example 2. HPLC: tR=1.52 min (Method A); MS-ES: (M+H)+=514.
The compound is prepared analogous to Example 2. HPLC: tR=1.78 min (Method A); MS-ES: (M+H)+=495.
The compound is prepared analogous to Example 2. HPLC: tR=1.35 min (Method A); MS-ES: (M+H)+=664.
The compound is prepared analogous to Example 2. HPLC: tR=1.36 min (Method A); MS-ES: (M+H)+=583.
The compound is prepared analogous to Example 2. HPLC: tR=1.14 min (Method A); MS-ES: (M+H)+=596.
The compound is prepared analogous to Example 2. HPLC: tR=1.18 min (Method A); MS-ES: (M+H)+=493.
The compound is prepared analogous to Example 2. HPLC: tR=0.98 min (Method A); MS-ES: (M+H)+=596.
The compound is prepared analogous to Example 2. HPLC: tR=0.93 min (Method A); MS-ES: (M+H)+=575.
The compound is prepared analogous to Example 2. HPLC: tR=0.96 min (Method A); MS-ES: (M+H)+=548.
The compound is prepared analogous to Example 2. HPLC: tR=1.28 min (Method A); MS-ES: (M+H)+=458.
The compound is prepared analogous to Example 2. HPLC: tR=1.73 min (Method A); MS-ES: (M+H)+=528.
The compound is prepared analogous to Example 2. HPLC: tR=1.07 min (Method A); MS-ES: (M+H)+=536.
The compound is prepared analogous to Example 2. HPLC: tR=0.92 min (Method A); MS-ES: (M+H)+=548.
The compound is prepared analogous to Example 2. HPLC: tR=0.94 min (Method A); MS-ES: (M+H)+=521.
The compound is prepared analogous to Example 2. HPLC: tR=0.94 min (Method A); MS-ES: (M+H)+=521.
The compound is prepared analogous to Example 2. HPLC: tR=0.93 min (Method A); MS-ES: (M+H)+=534.
The compound is prepared analogous to Example 2. HPLC: tR=1.17 min (Method A); MS-ES: (M+H)+=602.
The compound is prepared analogous to Example 2. HPLC: tR=1.08 min (Method A); MS-ES: (M+H)+=616.
The compound is prepared analogous to Example 2. HPLC: tR=1.09 min (Method A); MS-ES: (M+H)+=535.
The compound is prepared analogous to Example 2. HPLC: tR=0.89 min (Method A); MS-ES: (M+H)+=548.
The compound is prepared analogous to Example 2. HPLC: tR=1.43 min (Method A); MS-ES: (M+H)+=616.
The compound is prepared analogous to Example 2. HPLC: tR=1.10 min (Method A); MS-ES: (M+H)+=548.
The compound is prepared analogous to Example 2. HPLC: tR=1.17 min (Method A); MS-ES: (M+H)+=535.
The compound is prepared analogous to Example 2. HPLC: tR=1.20 min (Method A); MS-ES: (M+H)+=534.
The compound is prepared analogous to Example 2. HPLC: tR=1.16 min (Method A); MS-ES: (M+H)+=422.
The compound is prepared analogous to Example 2. HPLC: tR=0.95 min (Method A); MS-ES: (M+H)+=423.
The compound is prepared analogous to Example 2. HPLC: tR=0.93 min (Method A); MS-ES: (M+H)+=423.
The compound is prepared analogous to Example 2. HPLC: tR=0.96 min (Method A); MS-ES: (M+H)+=423.
The compound is prepared analogous to Example 3 (scheme 3). HPLC: tR=7.59 min. (Method B); MS-ES: (M+H)+=485; TLC**: Rf=0.17
The compound is prepared analogous to Example 3. HPLC: tR=1.78 min (Method A); MS-ES: (M+H)+=468.
The compound is prepared analogous to Example 2. HPLC: tR=1.60 min (Method A); MS-ES: (M+H)+=436.
The compound is prepared analogous to Example 2. HPLC: tR=1.24 min (Method A); MS-ES: (M+H)+=437.
The compound is prepared analogous to Example 2. HPLC: tR=1.22 min (Method A); MS-ES: (M+H)+=437.
The compound is prepared analogous to Example 2. HPLC: tR=1.24 min (Method A); MS-ES: (M+H)+=437.
The compound is prepared analogous to Example 2. HPLC: tR=1.15 min (Method A); MS-ES: (M+H)+=438.
The compound is prepared analogous to Example 2. HPLC: tR=1.26 min (Method A); MS-ES: (M+H)+=451.
The compound is prepared analogous to Example 2. HPLC: tR=1.39 min (Method A); MS-ES: (M+H)+=467.
The compound is prepared analogous to Example 2. HPLC: tR=1.05 min (Method A); MS-ES: (M+H)+=535.
The compound is prepared analogous to Example 2. HPLC: tR=1.33 min (Method A); MS-ES: (M+H)+=447.
The compound is prepared analogous to Example 2. HPLC: tR=1.21 min (Method A); MS-ES: (M+H)+=433.
The compound is prepared analogous to Example 2. HPLC: tR=1.17 min (Method A); MS-ES: (M+H)+=419.
The compound is prepared analogous to Example 2. HPLC: tR=1.29 min (Method A); MS-ES: (M+H)+=449.
The compound is prepared analogous to Example 2. HPLC: tR=1.52 min (Method A); MS-ES: (M+H)+=477.
The compound is prepared analogous to Example 2. HPLC: tR=1.91 min (Method A); MS-ES: (M+H)+=487.
The compound is prepared analogous to Example 2. HPLC: tR=1.71 min (Method A); MS-ES: (M+H)+=444.
The compound is prepared analogous to Example 2. HPLC: tR=1.15 min (Method A); MS-ES: (M+H)+=380.
The compound is prepared analogous to Example 2. HPLC: tR=1.26 min (Method A); MS-ES: (M+H)+=396.
The compound is prepared analogous to Example 2. HPLC: tR=1.57 min (Method A); MS-ES: (M+H)+=458.
The compound is prepared analogous to Example 2. HPLC: tR=1.42 min (Method A); MS-ES: (M+H)+=459.
The compound is prepared analogous to Example 2. HPLC: tR=1.64 min (Method A); MS-ES: (M+H)+=432.
The compound is prepared analogous to Example 2. HPLC: tR=1.20 min (Method A); MS-ES: (M+H)+=431.
The compound is prepared analogous to Example 2. HPLC: tR=1.55 min (Method A); MS-ES: (M+H)+=426.
The compound is prepared analogous to Example 2. HPLC: tR=1.76 min (Method A); MS-ES: (M+H)+=469.
The compound is prepared analogous to Example 2. HPLC: tR=1.58 min (Method A); MS-ES: (M+H)+=425.
The compound is prepared analogous to Example 2. HPLC: tR=1.13 min (Method A); MS-ES: (M+H)+=415.
The compound is prepared analogous to Example 2. HPLC: tR=1.36 min (Method A); MS-ES: (M+H)+=444.
The compound is prepared analogous to Example 2. HPLC: tR=1.57 min (Method A); MS-ES: (M+H)+=444.
The compound is prepared analogous to Example 2. HPLC: tR=1.18 min (Method A); MS-ES: (M+H)+=440.
The compound is prepared analogous to Example 2. HPLC: tR=1.27 min (Method A); MS-ES: (M+H)+=451.
The compound is prepared analogous to Example 2. HPLC: tR=1.13 min (Method A); MS-ES: (M+H)+=535.
The compound is prepared analogous to Example 2. MS-ES: (M+H)+=495.
The compound is prepared analogous to Example 2. HPLC: tR=1.13 min (Method A); MS-ES: (M+H)+=548.
The compound is prepared analogous to Example 2. HPLC: tR=1.26 min (Method A); MS-ES: (M+H)+=443.
The compound is prepared analogous to Example 2. HPLC: tR=0.85 min (Method A); MS-ES: (M+H)+=426.
The compound is prepared analogous to Example 2. HPLC: tR=0.97 min (Method A); MS-ES: (M+H)+=536.
The compound is prepared analogous to Example 2. HPLC: tR=1.09 min (Method A); MS-ES: (M+H)+=463.
The compound is prepared analogous to Example 2. HPLC: tR=1.15 min (Method A); MS-ES: (M+H)+=479.
The compound is prepared analogous to Example 2. HPLC: tR=1.23 min (Method A); MS-ES: (M+H)+=484.
The compound is prepared analogous to Example 2. HPLC: tR=0.90 min (Method A); MS-ES: (M+H)+=470.
The compound is prepared analogous to Example 2. HPLC: tR=1.05 min (Method A); MS-ES: (M+H)+=404.
The compound is prepared analogous to Example 2. HPLC: tR=0.92 min (Method A); MS-ES: (M+H)+=548.
The compound is prepared analogous to Example 2. HPLC: tR=1.11 min (Method A); MS-ES: (M+H)+=562.
The compound is prepared analogous to Example 2. HPLC: tR=0.90 min (Method A); MS-ES: (M+H)+=546.
The compound is prepared analogous to Example 2. HPLC: tR=1.08 min (Method A); MS-ES: (M+H)+=560.
The compound is prepared analogous to Example 2. HPLC: tR=0.87 min (Method A); MS-ES: (M+H)+=534.
The compound is prepared analogous to Example 2. HPLC: tR=1.11 min (Method A); MS-ES: (M+H)+=548.
The compound is prepared analogous to Example 2. HPLC: tR=0.93 min (Method A); MS-ES: (M+H)+=548.
The compound is prepared analogous to Example 2. HPLC: tR=1.12 min (Method A); MS-ES: (M+H)+=562.
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2, Final deprotection is achieved with TFA in CH2Cl2 at rt. MS-ES: (M+H)+=521.
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2, Final deprotection is achieved with TFA in CH2Cl2 at rt. MS-ES: (M+H)+=507.
The compound is prepared analogous to Example 2. HPLC: tR=4.37 min (Method C); MS-ES: (M+H)+=565.
The compound is prepared analogous to Example 2. HPLC: tR=3.91 min (Method C); MS-ES: (M+H)+=551.
The compound is prepared analogous to Example 2. HPLC: tR=4.37 min (Method C); MS-ES: (M+H)+=549.
The compound is prepared analogous to Example 2. HPLC: tR=3.99 min (Method C); MS-ES: (M+H)+=535.
The compound is prepared analogous to Example 2. HPLC: tR=4.43 min (Method C); MS-ES: (M+H)+=547.
The compound is prepared analogous to Example 2. HPLC: tR=4.46 min (Method C); MS-ES: (M+H)+=549.
The compound is prepared analogous to Example 2. HPLC: tR=3.99 min (Method C); MS-ES: (M+H)+=533.
The compound is prepared analogous to Example 2. HPLC: tR=4.02 min (Method C); MS-ES: (M+H)+=535.
The compound is prepared analogous to Example 2. HPLC: tR=4.23 min (Method C); MS-ES: (M+H)+=476.
The compound is prepared analogous to Example 2. HPLC: tR=4.29 min (Method C); MS-ES: (M+H)+=478.
The compound is prepared analogous to Example 2. HPLC: tR=4.30 min (Method C); MS-ES: (M+H)+=478.
The compound is prepared analogous to Example 2. HPLC: tR=4.43 min (Method C); MS-ES: (M+H)+=476.
The compound is prepared analogous to Example 2. MS-ES: (M+H)+=465.
The compound is prepared analogous to Example 3. HPLC: tR=3.20 min (Method D); MS-ES: (M+H)+=456.
The compound is prepared analogous to Example 3. HPLC: tR=3.21 min (Method D); MS-ES: (M+H)+=411.
The compound is prepared analogous to Example 3. HPLC: tR=2.80 min (Method D); MS-ES: (M+H)+=430.
The compound is prepared analogous to Example 3. HPLC: tR=2.83 min (Method D); MS-ES: (M+H)+=452.
The compound is prepared analogous to Example 2. HPLC: tR=4.61 min (Method C); MS-ES: (M+H)+=547.
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2, Final deprotection is achieved with 4N HCl in dioxan at rt. HPLC: tR=4.19 min (Method C); MS-ES: (M+H)+=450.
The compound is prepared analogous to Example 3. HPLC: tR=3.03 min (Method D); MS-ES: (M+H)+=469.
The compound is prepared analogous to Example 3. HPLC: tR=6.74 min (Method B); MS-ES: (M+H)+=553; TLC**: Rf=0.10
The compound is prepared analogous to Example 3. HPLC: tR=6.75 min. (Method B); MS-ES: (M+H)+=530; TLC**: Rf=0.21
The compound is prepared analogous to Example 3. But instead of using 2-cloro-7H-pyrrolo[2,3-d]pyrimidine, 2-chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidine is used in the first step. This starting material is obtained by using tributyl-(1-propynyl)tin in the Stille coupling in the synthesis for the pyrrolopyrimidine as exemplified in the reaction scheme of Example 1. HPLC: tR=8.46 min. (Method B); MS-ES: (M+H)+=582; TLC**: Rf=0.51
The compound is prepared analogous to Example 3. But instead of using 2-cloro-7H-pyrrolo[2,3-d]pyrimidine, 2-chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidine is used in the first step. This starting material is obtained by using tributyl-(1-propynyl)tin in the Stille coupling in the synthesis for the pyrrolopyrimidine as exemplified in the reaction scheme of Example 1. HPLC: tR=8.59 min. (Method B); MS-ES: (M+H)+=520; TLC‡: Rf=0.22
The compound is prepared analogous to Example 2. HPLC: tR=7.22 min. (Method B); MS-ES: (M+H)+=517; TLC*: Rf=0.37
The compound is prepared analogous to Example 3. HPLC: tR=7.85 min. (Method B); MS-ES: (M+H)+=450; TLC‡: Rf=0.41
The compound is prepared analogous to Example 3. HPLC: tR=8.88 min. (Method B); MS-ES: (M+H)+=498; TLC‡: Rf=0.32
The compound is prepared analogous to Example 3. HPLC: tR=8.49 min. (Method B); MS-ES: (M+H)+=494; TLC‡: Rf=0.28
The compound is prepared analogous to Example 3. HPLC: tR=7.37 min. (Method B); MS-ES: (M+H)+=606; TLC**: Rf=0.34
The compound is prepared analogous to Example 3. HPLC: tR=7.47 min. (Method B); MS-ES: (M+H)+=544; TLC**: Rf=0.35
The compound is prepared analogous to Example 3. HPLC: tR=8.66 min. (Method B); MS-ES: (M+H)+=442; TLC**: Rf=0.48
The compound is prepared analogous to Example 3. HPLC: tR=8.95 min. (Method B); MS-ES: (M+H)+=482; TLC**: Rf=0.46
The compound is prepared analogous to Example 3. HPLC: tR=8.29 min. (Method B); MS-ES: (M+H)+=462; TLC**: Rf=0.25
The compound is prepared analogous to Example 3. HPLC: tR=7.71 min. (Method B); MS-ES: (M+H)+=489; TLC**: Rf=0.33
The compound is prepared analogous to Example 3. HPLC: tR=7.37 min. (Method B); MS-ES: (M+H)+=502; TLC*: Rf=0.24
The compound is prepared analogous to Example 3. HPLC: tR=7.67 min. (Method B); MS-ES: (M+H)+=487; TLC*: Rf=0.30
The compound is prepared analogous to Example 3. HPLC: tR=7.27 min. (Method B); MS-ES: (M+H)+=471; TLC*: Rf=0.50
The compound is prepared analogous to Example 3. HPLC: tR=7.25 min. (Method B); MS-ES: (M+H)+=418; TLC*: Rf=0.55
The compound is prepared analogous to Example 3. HPLC: tR=7.23 min. (Method B); MS-ES: (M+H)+=422; TLC*: Rf=0.50
The compound is prepared analogous to Example 3. HPLC: tR=8.67 min. (Method B); MS-ES: (M+H)+=462; TLC*: Rf=0.59
The compound is prepared analogous to Example 3. HPLC: tR=7.97 min. (Method B); MS-ES: (M+H)+=463; TLC**: Rf=0.17
The compound is prepared analogous to Example 3. HPLC: tR=8.15 min. (Method B); MS-ES: (M+H)+=491; TLC**: Rf=0.32
The compound is prepared analogous to Example 3. HPLC: tR=8.50 min. (Method B); MS-ES: (M+H)+=489; TLC**: Rf=0.50
The compound is prepared analogous to Example 3. HPLC: tR=7.80 min. (Method B); MS-ES: (M+H)+=466; TLC*: Rf=0.77
The compound is prepared analogous to Example 3. HPLC: tR=8.81 min. (Method B); MS-ES: (M+H)+=514; TLC*: Rf=0.67
The compound is prepared analogous to Example 3. HPLC: tR=8.23 min. (Method B); MS-ES: (M+H)+=478; TLC*: Rf=0.80
The compound is prepared analogous to Example 3. HPLC: tR=8.05 min. (Method B); MS-ES: (M+H)+=477; TLC*: Rf=0.67
The compound is prepared analogous to Example 3. HPLC: tR=7.94 min. (Method B); MS-ES: (M+H)+=436; TLC*: Rf=0.37
The compound is prepared analogous to Example 3. HPLC: tR=7.81 min. (Method B); MS-ES: (M+H)+=463; TLC*: Rf=0.61
The compound is prepared analogous to Example 2. The final product is obtained by cleaving the Boc-protecting group under acidic conditions (4 M HCl in dioxane). HPLC: tR=6.83 min. (Method B); MS-ES: (M+H)+=516
The compound is prepared analogous to Example 2. The final product is obtained by cleaving the Boc-protecting group under acidic conditions (4 M HCl in dioxane). HPLC: tR=7.00 min. (Method B); MS-ES: (M+H)+=534
The compound is prepared analogous to Example 2. The final product is obtained by cleaving the Boc-protecting group under acidic conditions (4 M HCl in dioxane). HPLC: tR=6.60 min. (Method B); MS-ES: (M+H)+=520
The compound is prepared analogous to Example 2. HPLC: tR=6.42 min. (Method B); MS-ES: (M+H)+=549; TLC*: Rf=0.11
The compound is prepared analogous to Example 2. HPLC: tR=6.91 min. (Method B); MS-ES: (M+H)+=492
The compound is prepared analogous to Example 2. HPLC: tR=6.61 min. (Method B); MS-ES: (M+H)+=562; TLC*: Rf=0.25
The compound is prepared analogous to Example 2. HPLC: tR=7.01 min. (Method B); MS-ES: (M+H)+=505; TLC*: Rf=0.21
The compound is prepared analogous to Example 2. HPLC: tR=8.04 min. (Method B); MS-ES: (M+H)+=595; TLC*: Rf=0.43
The compound is prepared from Example 320 by cleaving the Boc-protecting group under acidic conditions (4 M HCl in dioxane). HPLC: tR=6.13 min. (Method B); MS-ES: (M+H)+=495
The compound is prepared analogous to Example 2. HPLC: tR=7.23 min. (Method B); MS-ES: (M+H)+=558; TLC*: Rf=0.25
The compound is prepared analogous to Example 2. HPLC: tR=7.46 min. (Method B); MS-ES: (M+H)+=576; TLC*: Rf=0.18
The compound is prepared analogous to Example 3. HPLC: tR=8.47 min. (Method B); MS-ES: (M+H)+=432
The compound is prepared analogous to Example 3. HPLC: tR=7.60 min. (Method B); MS-ES: (M+H)+=436; TLC*: Rf=0.77
The compound is prepared analogous to Example 2. HPLC: tR=7.30 min. (Method B); MS-ES: (M+H)+=544; TLC*: Rf=0.35
The compound is prepared analogous to Example 2. HPLC: tR=7.44 min. (Method B); MS-ES: (M+H)+=531; TLC*: Rf=0.48
The compound is prepared analogous to Example 2. HPLC: tR=7.32 min. (Method B); MS-ES: (M+H)+=478; TLC*: Rf=0.56
The compound is prepared analogous to Example 2. HPLC: tR=6.75 min. (Method B); MS-ES: (M+H)+=560; TLC*: Rf=0.23
The compound is prepared analogous to Example 2. HPLC: tR=7.16 min. (Method B); MS-ES: (M+H)+=556; TLC*: Rf=0.30
The compound is prepared analogous to Example 2. HPLC: tR=6.72 min. (Method B); MS-ES: (M+H)+=548; TLC*: Rf=0.35
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2, Final deprotection is achieved with 4 M HCl in dioxane at rt. HPLC: tR=6.45 min. (Method B); MS-ES: (M+H)+=534
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2, Final deprotection is achieved with 4 M HCl in dioxane at rt. HPLC: tR=6.90 min. (Method B); MS-ES: (M+H)+=477
The compound is prepared analogous to Example 2. HPLC: tR=6.55 min. (Method B); MS-ES: (M+H)+=548; TLC*: Rf=0.39
The compound is prepared analogous to Example 2. HPLC: tR=7.04 min. (Method B); MS-ES: (M+H)+=491; TLC**: Rf=0.10
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2, Final deprotection is achieved with 4 M HCl in dioxane at rt. HPLC: tR=7.10 min. (Method B); MS-ES: (M+H)+=530
The compound is prepared analogous to Example 2. HPLC: tR=7.19 min. (Method B); MS-ES: (M+H)+=544; TLC*: Rf=0.27
The compound is prepared analogous to Example 2. HPLC: tR=7.02 min. (Method B); MS-ES: (M+H)+=540; TLC*: Rf=0.24
The compound is prepared analogous to Example 2. HPLC: tR=6.70 min. (Method B); MS-ES: (M+H)+=575; TLC*: Rf=0.15
The compound is prepared analogous to Example 2. HPLC: tR=6.76 min. (Method B); MS-ES: (M+H)+=573; TLC*: Rf=0.15
The compound is prepared analogous to Example 2. HPLC: tR=6.47 min. (Method B); MS-ES: (M+H)+=533
The compound is prepared analogous to Example 2. HPLC: tR=6.42 min. (Method B); MS-ES: (M+H)+=548
The compound is prepared analogous to Example 2. HPLC: tR=6.77 min. (Method B); MS-ES: (M+H)+=561; TLC*: Rf=0.18
The compound is prepared analogous to Example 2. HPLC: tR=6.88 min. (Method B); MS-ES: (M+H)+=548; TLC*: Rf=0.35
The compound is prepared analogous to Example 2. HPLC: tR=8.02 min. (Method B); MS-ES: (M+H)+=560; TLC*: Rf=0.18
The compound is prepared analogous to Example 2. The corresponding Boc-protected aniline is obtained from the procedure described in the J. Am. Chem. Soc. 2006, 128, 3538-3539 starting from 4-bromo-3-methyl-phenylamine. HPLC: tR=0.58 min (Method G); MS-ES: (M+H)+=505
The compound is prepared analogous to Example 2, using a TBDPS-protected alcohol derivative in Step 2.2, Final deprotection is achieved with 1 M TBAF in THF at rt. HPLC: tR=0.57 min (Method G); MS-ES: (M+H)+=508
The compound is prepared analogous to Example 2. HPLC: tR=7.82 min. (Method B); MS-ES: (M+H)+=520; TLC*: Rf=0.12
The compound is prepared analogous to Example 2. HPLC: tR=6.71 min. (Method B); MS-ES: (M+H)+=464; TLC*: Rf=0.03
The compound is prepared analogous to 2, HPLC: tR=0.67 min (Method G); MS-ES: (M+H)+=523
The compound is prepared analogous to Example 2. HPLC: tR=7.77 min. (Method B); MS-ES: (M+H)+=535; TLC*: Rf=0.05
The compound is prepared analogous to Example 2. HPLC: tR=0.58 min (Method G); MS-ES: (M+H)+=552
The compound is prepared analogous to Example 2. HPLC: tR=8.07 min. (Method B); MS-ES: (M+H)+=548; TLC*: Rf=0.22
The compound is prepared analogous to Example 2, using a Boc-protected morpholine derivative in Step 2.2, Final deprotection is achieved with 1 M HCl in EtOH at 60° C. HPLC: tR=0.59 min (Method G); MS-ES: (M+H)+=507
The compound is prepared analogous to Example 2. HPLC: tR=6.73 min. (Method B); MS-ES: (M+H)+=562; TLC*: Rf=0.22
The compound is prepared analogous to Example 2. HPLC: tR=7.77 min. (Method B); MS-ES: (M+H)+=535; TLC*: Rf=0.04
The compound is prepared analogous to Example 2. HPLC: tR=6.64 min. (Method B); MS-ES: (M+H)+=479; TLC*: Rf=0.14
The compound is prepared analogous to Example 2. HPLC: tR=6.93 min. (Method B); MS-ES: (M+H)+=577; TLC*: Rf=0.58
The compound is prepared analogous to Example 2. HPLC: tR=7.60 min. (Method B); MS-ES: (M+H)+=573; TLC*: Rf=0.51
The compound is prepared analogous to Example 2. HPLC: tR=6.65 min. (Method B); MS-ES: (M+H)+=577; TLC (15% methanol/85% methylene chloride): Rf=0.16
The compound is prepared analogous to Example 2. HPLC: tR=6.47 min. (Method B); MS-ES: (M+H)+=549; TLC (15% methanol/85% methylene chloride): Rf=0.15
The compound is prepared analogous to Example 2. HPLC: tR=6.95 min. (Method B); MS-ES: (M+H)+=578; TLC (15% methanol/85% methylene chloride): Rf=0.47
The compound is prepared analogous to Example 2. HPLC: tR=8.03 min. (Method B); MS-ES: (M+H)+=535; TLC*: Rf=0.10
The compound is prepared analogous to Example 2. HPLC (racemic mixture): tR=3.62 min and 4.37 min (Method C); MS-ES: (M+H)+=575
The compound is prepared analogous to Example 2. HPLC: tR=6.94 min. (Method B); MS-ES: (M+H)+=574; TLC*: Rf=0.38
The compound is prepared analogous to Example 2. HPLC: tR=7.62 min. (Method B); MS-ES: (M+H)+=570; TLC*: Rf=0.55
The compound is prepared analogous to Example 2. HPLC: tR=7.41 min (Method B); MS-ES: (M+H)+=558
The compound is prepared analogous to Example 2. HPLC: tR=6.84 min (Method B); MS-ES: (M+H)+=492; TLC*: Rf=0.03
The compound is prepared analogous to Example 2. HPLC: tR=6.67 min (Method B); MS-ES: (M+H)+=479; TLC (50% methanol/50% methylene chloride): Rf=0.26
The compound is obtained by treating [7-(3,5-difluoro-4-morpholin-4-ylmethyl-phenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-(4-methyl-3-piperazin-1-ylmethyl-phenyl)-amine (Example 332) with formic acid 4-nitro-phenyl ester in presence of triethylamine in THF. HPLC: tR=6.84 min (Method B); MS-ES: (M+H)+=562
The compound is prepared analogous to Example 2. HPLC: tR=6.53 min (Method B); MS-ES: (M+H)+=563; TLC (15% methanol/85% methylene chloride): Rf=0.28
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with 4 M HCl in dioxane at rt. HPLC: tR=6.52 min (Method B); MS-ES: (M+H)+=521; TLC (64% methanol/32% methylene chloride/4% aq. ammonia 24% ): Rf=0.49
The compound is prepared analogous to Example 2. HPLC: tR=6.62 min (Method B); MS-ES: (M+H)+=560
The compound is prepared analogous to Example 2. HPLC: tR=7.33 min (Method B); MS-ES: (M+H)+=556
The compound is prepared analogous to Example 2. HPLC: tR=7.26 min (Method B); MS-ES: (M+H)+=521; TLC (50% methanol/50% methylene chloride): Rf=0.30
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with 4 M HCl in dioxane at rt. HPLC: tR=7.86 min (Method B); MS-ES: (M+H)+=534; TLC (50% methanol/50% methylene chloride): Rf=0.09
The compound is prepared analogous to Example 2. HPLC: tR=3.90 min (Method C); MS-ES: (M+H)+=521
The compound is prepared analogous to Example 2. HPLC: tR=6.73 min (Method B); MS-ES: (M+H)+=562; TLC (100% methanol): Rf=0.11
The compound is prepared analogous to Example 2. HPLC: tR=7.38 min (Method B); MS-ES: (M+H)+=558; TLC (100% methanol): Rf=0.12
The compound is prepared analogous to Example 2. HPLC: tR=6.89 min (Method B); MS-ES: (M+H)+=493; TLC (50% methanol/50% methylene chloride): Rf=0.25
The compound is prepared analogous to Example 2, using a trifluoroacetyl-protected piperidine derivative in Step 2.2. Final deprotection is achieved with K2CO3 in methanol/water at rt. The corresponding aniline is obtained by the procedure described in patent WO98/05292 p. 104-105 compound (111), followed by nitro reduction. HPLC: tR=7.52 min (Method B); MS-ES: (M+H)+=515; TLC (50% methanol/50% methylene chloride): Rf=0.06
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with 4 M HCl in dioxane at rt. HPLC: tR=6.64 min (Method B); MS-ES: (M+H)+=547
The compound is prepared analogous to Example 2, using a trifluoroacetyl-protected piperidine derivative in Step 2.2. Final deprotection is achieved with K2CO3 in methanol/water at rt. The corresponding aniline is obtained by the procedure described in patent WO98/05292 p. 104-105 compound (111), followed by nitro reduction. HPLC: tR=6.93 min (Method B); MS-ES: (M+H)+=519; TLC (49% methanol/49% methylene chloride/2% 7N ammonia in methanol): Rf=0.06
The compound is prepared analogous to Example 2. HPLC: tR=6.57 min (Method B); MS-ES: (M+H)+=535
The compound is prepared analogous to Example 2. HPLC: tR=7.03 min (Method B); MS-ES: (M+H)+=562
The compound is prepared analogous to Example 2. HPLC: tR=7.82 min (Method B); MS-ES: (M+H)+=558
The compound is prepared analogous to Example 2. HPLC: tR=8.06 min (Method B); MS-ES: (M+H)+=562; TLC (33% methanol/67% methylene chloride): Rf=0.23
The compound is prepared analogous to Example 2. HPLC: tR=4.00 min (Method C); MS-ES: (M)+=521
The compound is prepared analogous to Example 2. HPLC: tR=0.60 min (Method G); MS-ES: (M+H)+=574
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with 4 M HCl in dioxane at rt. HPLC: tR=6.37 min (Method B); MS-ES: (M+H)+=535
The compound is prepared analogous to Example 2. HPLC: tR=6.75 min (Method B); MS-ES: (M+H)+=549
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with 4 M HCl in dioxane at rt. HPLC: tR=6.67 min (Method B); MS-ES: (M+H)+=536; TLC (33% methanol/66% methylene chloride/1% aq. ammonia 24%): Rf=0.28
The compound is prepared analogous to Example 2. HPLC: tR=7.08 min (Method B); MS-ES: (M+H)+=555; TLC*: Rf=0.63
The compound is prepared analogous to Example 2. HPLC: tR=6.67 min (Method B); MS-ES: (M+H)+=521; TLC (20% methanol/80% methylene chloride): Rf=0.36
The compound is prepared analogous to Example 2. HPLC: tR=6.83 min (Method B); MS-ES: (M+H)+=562
The compound is prepared analogous to Example 2. HPLC: tR=6.51 min (Method B); MS-ES: (M+H)+=592
The compound is prepared analogous to Example 2. HPLC: tR=7.00 min (Method B); MS-ES: (M+H)+=579
The compound is prepared analogous to Example 2. HPLC: tR=6.81 min (Method B); MS-ES: (M+H)+=605
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with 4 M HCl in dioxane at rt. HPLC: tR=6.76 min (Method B); MS-ES: (M+H)+=548; TLC*: Rf=0.27
The compound is prepared analogous to Example 2. HPLC: tR=7.04 min (Method B); MS-ES: (M+H)+=541
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with 4 M HCl in dioxane at rt. HPLC: tR=6.93 min (Method B); MS-ES: (M+H)+=554
The compound is prepared analogous to Example 2. HPLC: tR=7.71 min (Method B); MS-ES: (M+H)+=542
The compound is prepared analogous to Example 2. HPLC: tR=7.74 min (Method B); MS-ES: (M+H)+=646; TLC*: Rf=0.41
The compound is prepared analogous to Example 2. HPLC: tR=7.25 min (Method B); MS-ES: (M+H)+=602; TLC*: Rf=0.31
The compound is prepared analogous to Example 2. HPLC: tR=7.02 min (Method B); MS-ES: (M+H)+=535; TLC*: Rf=0.19
The compound is prepared analogous to Example 2. HPLC: tR=7.18 min (Method B); MS-ES: (M+H)+=579
The compound is prepared analogous to Example 2. HPLC: tR=6.81 min (Method B); MS-ES: (M+H)+=561
The compound is prepared analogous to Example 2. HPLC: tR=6.85 min (Method B); MS-ES: (M+H)+=589
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with 4 M HCl in dioxane at rt. HPLC: tR=6.38 min (Method B); MS-ES: (M+H)+=535
The compound is prepared analogous to Example 2. HPLC: tR=6.86 min (Method B); MS-ES: (M+H)+=619; TLC*: Rf=0.14
The compound is prepared analogous to Example 2. HPLC: tR=6.81 min (Method B); MS-ES: (M+H)+=535; TLC* with 1% aq. ammonia 24%: Rf=0.66
The compound is prepared analogous to Example 2. HPLC: tR=6.90 min (Method B); MS-ES: (M+H)+=516; TLC*: Rf=0.52
The compound is prepared analogous to Example 2. HPLC: tR=7.81 min (Method B); MS-ES: (M+H)+=516; TLC*: Rf=0.77
The compound is prepared analogous to Example 2. HPLC: tR=7.30 min (Method B); MS-ES: (M+H)+=517; TLC*: Rf=0.40
The compound is obtained from the reaction of 1-{5-[7-(3,5-Difluoro-4-morpholin-4-ylmethyl-phenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino]-2-methyl-benzyl}-piperazin-2-one (Example 399) with formaldehyde and sodium cyanoborohydride in methanol and DCM 2:1 at rt. HPLC: tR=6.91 min (Method B); MS-ES: (M+H)+=562; TLC*: Rf=0.28
The compound is prepared analogous to Example 2. HPLC: tR=6.45 min (Method B); MS-ES: (M+H)+=522
The compound is prepared analogous to Example 2. HPLC: tR=7.22 min (Method B); MS-ES: (M+H)+=549
The compound is prepared analogous to Example 2. HPLC: tR=7.11 min (Method B); MS-ES: (M+H)+=574; TLC*: Rf=0.19
The compound is prepared analogous to Example 2. HPLC: tR=7.19 min (Method B); MS-ES: (M+H)+=547; TLC*: Rf=0.31
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with 4 M HCl in dioxane at rt. HPLC: tR=6.98 min (Method B); MS-ES: (M+H)+=521
The compound is prepared analogous to Example 2. HPLC: tR=7.63 min (Method B); MS-ES: (M+H)+=596; TLC*: Rf=0.17
The compound is prepared analogous to Example 2. HPLC: tR=7.71 min (Method B); MS-ES: (M+H)+=610; TLC*: Rf=0.23
The compound is prepared analogous to Example 2. HPLC: tR=7.55 min (Method B); MS-ES: (M+H)+=569; TLC*: Rf=0.14
The compound is prepared analogous to Example 2, using a Boc-protected derivative in Step 2.2. Final deprotection is achieved with 4 M HCl in dioxane and then TFA at rt. The aniline building block is obtained from patent WO 2007/072158 p. 45 Scheme 4. HPLC: tR=6.78 min (Method B); MS-ES: (M+H)+=489; TLC (33% methanol/67% methylene chloride): Rf=0.52
The compound is prepared analogous to Example 2. HPLC: tR=7.68 min (Method B); MS-ES: (M+H)+=583; TLC*: Rf=0.10
The compound is prepared analogous to Example 2. HPLC: tR=6.77 min (Method B); MS-ES: (M+H)+=536
The compound is prepared analogous to Example 2. HPLC: tR=6.79 min (Method B); MS-ES: (M+H)+=549
The compound is prepared analogous to Example 2, using a Boc-protected derivative in Step 2.2. Final deprotection is achieved with 4 M HCl in dioxane at rt. HPLC: tR=7.43 min (Method B); MS-ES: (M+H)+=582; TLC*: Rf=0.15
The compound is prepared analogous to Example 2. HPLC: tR=7.74 min (Method B); MS-ES: (M+H)+=597; TLC*: Rf=0.06
The compound is prepared analogous to Example 2. HPLC: tR=8.33 min (Method B); MS-ES: (M+H)+=617; TLC*: Rf=0.15
The compound is prepared analogous to Example 2 with the aniline derivative obtained from the hydrogenation of 6-nitro-2,3-dihydro-1H-indole. HPLC: tR=6.71 min (Method B); MS-ES: (M+H)+=463; TLC**: Rf=0.24
The compound is prepared analogous to Example 2. HPLC: tR=7.86 min (Method B); MS-ES: (M+H)+=627
The compound is prepared analogous to Example 2. HPLC: tR=7.04 min (Method B); MS-ES: (M+H)+=579
The compound is prepared analogous to Example 2. HPLC: tR=6.78 min (Method B); MS-ES: (M+H)+=562
The compound is prepared analogous to Example 2. HPLC: tR=7.57 min (Method B); MS-ES: (M+H)+=610
The compound is prepared analogous to Example 2. HPLC: tR=7.37 min (Method B); MS-ES: (M+H)+=545
The compound is prepared analogous to Example 2. HPLC: tR=7.10 min (Method B); MS-ES: (M+H)+=526; TLC*: Rf=0.23
The compound is prepared analogous to Example 3. HPLC: tR=4.79 min (Method F); MS-ES: (M+H)+=414
The compound is prepared analogous to Example 3. HPLC: tR=4.45 min (Method F); MS-ES: (M+H)+=425
The compound is prepared analogous to Example 2. HPLC: tR=4.29 min (Method F); MS-ES: (M+H)+=569
The compound is prepared analogous to Example 2. HPLC: tR=4.40 min (Method F); MS-ES: (M+H)+=583; TLC (20% methanol/80% methylene chloride): Rf=0.32
The compound is prepared analogous to Example 2. HPLC: tR=4.48 min (Method F); MS-ES: (M+H)+=581; TLC (20% methanol/80% methylene chloride): Rf=0.38
The compound is prepared analogous to Example 2. HPLC: tR=4.36 min (Method F); MS-ES: (M+H)+=569
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with TFA in CH2Cl2 at rt. HPLC: tR=4.29 min (Method F); MS-ES: (M+H)+=555
The compound is prepared analogous to Example 3. HPLC: tR=4.74 min (Method F); MS-ES: (M+H)+=447
The compound is prepared analogous to Example 3. The corresponding aryle bromide is obtained from the reaction of (4-bromo-2,6-difluoro-phenyl)-methanol withiodomethane in presence of sodium hydride in THF. HPLC: tR=4.69 min (Method F); MS-ES: (M+H)+=466
The compound is prepared analogous to Example 2. HPLC: tR=4.14 min (Method F); MS-ES: (M+H)+=549
The compound is prepared analogous to Example 3. HPLC: tR=4.25 min (Method F); MS-ES: (M+H)+=455
The compound is prepared analogous to Example 3. The corresponding aryle bromide is obtained from the reaction of 1-(4-bromo-phenyl)-ethanol on iodomethane under Ar in presence of sodium hydride in THF. HPLC: tR=4.54 min (Method F); MS-ES: (M+H)+=444
The compound is prepared analogous to Example 2. HPLC: tR=6.71 min (Method B); MS-ES: (M+H)+=588; TLC (53% chloroform, 36% methanol, 10% water, 0.5% acetic acid): Rf=0.25
The compound is prepared analogous to Example 2. HPLC: tR=3.94 min (Method C); MS-ES: (M+H)+=562
The compound is prepared analogous to Example 2. HPLC: tR=4.00 min (Method C); MS-ES: (M+H)+=576
The compound is prepared analogous to Example 2. HPLC: tR=0.74 min (Method El); MS-ES: (M+H)+=436
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with TFA in CH2Cl2 at rt. HPLC: tR=1.14 min (Method E2); MS-ES: (M+H)+=535
The compound is prepared analogous to Example 2. HPLC: tR=3.97 min (Method C); MS-ES: (M+H)+=562
The compound is prepared analogous to Example 2. HPLC: tR=3.96 min (Method C); MS-ES: (M+H)+=547
The compound is prepared analogous to Example 2. HPLC: tR=3.89 min (Method C); MS-ES: (M+H)+=547
The compound is prepared analogous to Example 2. HPLC: tR=0.81 min (Method E1); MS-ES: (M+H)+=533
The compound is prepared analogous to Example 2. HPLC: tR=4.02 min (Method C); MS-ES: (M+H)+=574
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with TFA in CH2Cl2 at rt. HPLC: tR=3.96 min (Method C); MS-ES: (M+H)+=534
The compound is prepared analogous to Example 2. HPLC: tR=4.01 min (Method C); MS-ES: (M+H)+=562
The compound is prepared analogous to Example 2. HPLC: tR=0.70 min (Method E1); MS-ES: (M+H)+=546
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with TFA in CH2Cl2 at rt. HPLC: tR=0.72 min (Method E1); MS-ES: (M+H)+=532
The compound is prepared analogous to Example 2. HPLC: tR=0.76 min (Method E1); MS-ES: (M+H)+=548
The compound is prepared analogous to Example 2. HPLC: tR=3.84 min (Method C); MS-ES: (M+H)+=578
The compound is prepared analogous to Example 2. HPLC: tR=3.99 min (Method F); MS-ES: (M+H)+=549
The compound is prepared analogous to Example 2. HPLC: tR=7.34 min (Method B); MS-ES: (M+H)+=560; TLC*: Rf=0.36
The compound is prepared analogous to Example 2. HPLC: tR=0.49 min (Method E1); MS-ES: (M+H)+=482
The compound is prepared analogous to Example 2. HPLC: tR=4.06 min (Method C); MS-ES: (M+H)+=575
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with TFA in CH2Cl2 at rt. HPLC: tR=3.91 min (Method C); MS-ES: (M+H)+=521
The compound is prepared analogous to Example 2. HPLC: tR=4.19 min (Method F); MS-ES: (M+H)+=541
The compound is prepared analogous to Example 2. HPLC: tR=1.12 min (Method E2); MS-ES: (M+H)+=535
The compound is prepared analogous to Example 2. HPLC: tR=0.01 min (Method C); MS-ES: (M+H)+=548
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with TFA in CH2Cl2 at rt. The aniline used for this synthesis is prepared from 2-(4-nitro-phenyl)-piperazine (racemic) via Boc-protection, nitro reduction and chiral separation (other enantiomer used for Example 475). HPLC: tR=0.68 min (Method E2); MS-ES: (M+H)+=506
The compound is prepared analogous to Example 2 and represents the enantiomer of Example 474. HPLC: tR=0.68 min (Method E2); MS-ES: (M+H)+=506
The compound is prepared analogous to Example 2. HPLC: tR=4.24 min (Method C); MS-ES: (M+H)+=588
The compound is prepared analogous to Example 2. HPLC: tR=4.06 min (Method C); MS-ES: (M+H)+=566
The compound is obtained from the reaction of 1-{5-[7-(3,5-Difluoro-4-morpholin-4-ylmethyl-phenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino]-pyridin-2-yl}-piperazin-2-one (Example 470) with formaldehyde and sodium cyanoborohydride in methanol and DCM 2:1 at rt. HPLC: tR=3.94 min (Method C); MS-ES: (M+H)+=535
The compound is prepared analogous to Example 2. HPLC: tR=3.97 min (Method C); MS-ES: (M+H)+=507
The compound is prepared analogous to Example 2. HPLC: tR=1.20 min (Method E2); MS-ES: (M+H)+=522
The compound is prepared analogous to Example 2. HPLC: tR=8.11 min (Method B); MS-ES: (M+H)+=530; TLC*: Rf=0.67
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with TFA in CH2Cl2 at rt. HPLC: tR=4.05 min (Method F); MS-ES: (M+H)+=534
The compound is prepared analogous to Example 2. HPLC: tR=4.58 min (Method F); MS-ES: (M+H)+=563
The compound is prepared analogous to Example 2. HPLC: tR=4.40 min (Method F); MS-ES: (M+H)+=562
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with TFA in CH2Cl2 at rt. HPLC: tR=4.44 min (Method F); MS-ES: (M+H)+=548
The compound is prepared analogous to Example 2. HPLC: tR=1.12 min (Method E); MS-ES: (M+H)+=521
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with TFA in CH2Cl2 at rt. HPLC: tR=3.99 min (Method C); MS-ES: (M+H)+=521
The compound is prepared analogous to Example 2. HPLC: tR=4.16 min (Method C); MS-ES: (M+H)+=549
The compound is prepared analogous to Example 2. HPLC: tR=4.42 min (Method F); MS-ES: (M+H)+=569
The compound is prepared analogous to Example 2. HPLC: tR=4.34 min (Method C); MS-ES: (M+H)+=560
The compound is prepared analogous to Example 2. HPLC: tR=4.56 min (Method C); MS-ES: (M+H)+=555
The compound is prepared analogous to Example 2. HPLC: tR=4.04 min (Method C); MS-ES: (M+H)+=549
The compound is prepared analogous to Example 2. HPLC: tR=3,54 min (Method C); MS-ES: (M+H)+=563
The compound is prepared analogous to Example 2. HPLC: tR=7.25 min (Method B); MS-ES: (M+H)+=568
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with TFA in CH2Cl2 at rt. HPLC: tR=3.98 min (Method C); MS-ES: (M+H)+=535
The compound is prepared analogous to Example 2. HPLC: tR=4.67 min (Method C); MS-ES: (M+H)+=603
The compound is prepared analogous to Example 2. HPLC: tR=4.07 min (Method C); MS-ES: (M+H)+=565
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with TFA in CH2Cl2 at rt. HPLC: tR=3.96 min (Method C); MS-ES: (M+H)+=534
The compound is prepared analogous to Example 2. HPLC: tR=4.27 min (Method C); MS-ES: (M+H)+=531
The compound is prepared analogous to Example 2. HPLC: tR=4.20 min (Method C); MS-ES: (M+H)+=521
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with TFA in CH2Cl2 at rt. HPLC: tR=3.92 min (Method C); MS-ES: (M+H)+=506
The compound is prepared analogous to Example 2, using a Boc-protected piperazine derivative in Step 2.2. Final deprotection is achieved with TFA in CH2Cl2 at rt. HPLC: tR=4.27 min (Method C); MS-ES: (M+H)+=541
The compound is prepared analogous to Example 2. HPLC: tR=4.00 min (Method C); MS-ES: (M+H)+=549
The compound is prepared analogous to Example 2. HPLC: tR=4.23 min (Method C); MS-ES: (M+H)+=535
The compound is prepared analogous to Example 2. HPLC: tR=4.06 min (Method C); MS-ES: (M+H)+=549
The compound is prepared analogous to Example 2. HPLC: tR=4.17 min (Method C); MS-ES: (M+H)+=535
The compound is prepared analogous to Example 2, using a Boc-protected morpholine derivative in Step 2.2. Final deprotection is achieved with HCl in EtOH at 60° C. HPLC: tR=0.60 min (Method G); MS-ES: (M+H)+=507
The compound is obtained by treating [7-(3,5-difluoro-4-morpholin-4-ylmethyl-phenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-((S)-3-morpholin-3-yl-phenyl)-amine (Example 507) with acetaldehyde in methanol and sodium cyanoborohydride at rt to 60° C. HPLC: tR=0.63 min (Method G); MS-ES: (M+H)+=535
The compound is obtained by treating [7-(3,5-difluoro-4-morpholin-4-ylmethyl-phenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-((S)-3-morpholin-3-yl-phenyl)-amine (Example 507) with formaldehyde in methanol and sodium cyanoborohydride at rt to 60° C. HPLC: tR=0.61 min (Method G); MS-ES: (M+H)+=521
The compound is prepared analogous to Example 2. HPLC: tR=0.61 min (Method G); MS-ES: (M+H)+=549; TLC (10% methanol/90% methylene chloride/1% ammonia): Rf=0.18
The compound is prepared analogous to Example 2, using a Boc-protected morpholine derivative in Step 2.2. Final deprotection is achieved with 1.25 M HCl in EtOH at 60° C. HPLC: tR=0.61 min (Method G); MS-ES: (M+H)+=507; TLC (10% methanol/90% methylene chloride/1% ammonia): Rf=0.50
The compound is prepared analogous to Example 2. HPLC: tR=0.65 min (Method G); MS-ES: (M+H)+=571
The compound is prepared analogous to Example 2. HPLC: tR=0.59 min (Method G); MS-ES: (M+H)+=552
The compound is prepared analogous to Example 2, using a Boc-protected azetidine derivative in Step 2.2. Final deprotection is achieved with 1.25 M HCl in EtOH at 60° C. HPLC: tR=0.55 min (Method G); MS-ES: (M+H)+=538
The compound is prepared analogous to Example 2. HPLC: tR=0.71 min (Method G); MS-ES: (M+H)+=523
The compound is prepared analogous to Example 2. HPLC: tR=0.67 min (Method G); MS-ES: (M+H)+=497
The compound is prepared analogous to Example 2. HPLC: tR=0.74 min (Method G); MS-ES: (M+H)+=537
The compound is prepared analogous to Example 2, using a TBDPS-protected alcohol derivative in Step 2.2. Final deprotection is achieved with 1 M TBAF in THF at rt. HPLC: tR=0.60 min (Method G) and MS-ES: (M+H)+=483.
The compound is prepared analogous to Example 2, using a Boc-protected pyrrolidine derivative in Step 2.2. Final deprotection is achieved with 1.25 M HCl in EtOH at 50° C. HPLC: tR=0.58 min (Method G); MS-ES: (M+H)+=508
The compound is obtained by treating [7-(3,5-difluoro-4-morpholin-4-ylmethyl-phenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-((S)-4-morpholin-3-yl-phenyl)-amine (Example 511) with benzyloxyacetaldehyde in methanol and sodium cyanoborohydride at 60° C. HPLC: tR=0.78 min (Method G); MS-ES: (M+H)+=641
The compound is obtained by treating [7-(3,5-Difluoro-4-morpholin-4-ylmethyl-phenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-((S)-4-morpholin-3-yl-phenyl)-amine (Example 511) with (tert-butyldimethylsilyloxy)-acetaldehyde in methanol and sodium cyanoborohydride at 60° C., followed by a deprotection with TBAF in THF. HPLC: tR=0.60 min (Method G); MS-ES: (M+H)+=551
The compound is prepared analogous to Example 3. HPLC: tR=1.60 min (Method A); MS-ES: (M+H)+=476.
5000 soft gelatin capsules, each comprising as active ingredient 0.05 g of one of the compounds of formula I mentioned in the preceding Examples, are prepared as follows: 250 g pulverized active ingredient is suspended in 2 L Lauroglykol® (propylene glycol laurate, Gattefossé S.A., Saint Priest, France) and ground in a wet pulverizer. 0.419 g portions of the mixture are then introduced into soft gelatin capsules using a capsule-filling machine.
Assays
Compounds of the present invention are assayed to measure their capacity to inhibit JAK2 and either STAT1 or STAT5 translocation pathway as described above. Results are provided in the Table 2:
Compounds of the invention selectively inhibit JAK2 when compared to other kinases, for example cMet, cKit, ALK and PDGFRa as shown by the results of Table 3.
The compounds are assayed in an antibody based kinase phosphorylation assay using the recombinant domains of cMet, cKit, ALK and PDGFRalpha kinases and a generic phospho-tyrosine peptide substrate. LanthaScreen™ is the detection of Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) using lanthanide chelates to measure interactions between various binding partners. In a TR-FRET kinase assay, a long-lifetime lanthanide donor species is conjugated to an antibody that specifically binds to a phosphorylated product of a kinase reaction that is labeled with a suitable acceptor fluorophore. This antibody-mediated interaction brings the lanthanide donor and the acceptor into proximity such that resonance energy transfer can take place, resulting in a detectible increase in FRET. The LanthaScreen is run at ambient temperature and terminated by the addition of stop solution, followed by Tb-labeled P-20 antibody. After incubating in the dark, the plates are transferred to a fluorescence reader for counting. The effect of compound on the enzymatic activity is in all assays obtained from the linear progress curves and determined from one reading (end point measurement).
Compounds of the present invention are further assayed to measure their capacity to inhibit JAK1, as described above. Compounds of ex. 1-522 show in this assay IC50-values in the range of <0.003->10 μmol l−1.
Compounds of the present invention are further assayed to measure their capacity to inhibit JAK3 as described above. Compounds of ex. 1-522 show in this assay IC50-values in the range of <0.003->10 μmol l−1.
Compounds of the present invention are further assayed to measure their capacity to inhibit TYK2 as described above. Compounds of ex. 1-522 show in this assay IC50-values in the range of <0.003->10 μmol l−1.
| Number | Date | Country | Kind |
|---|---|---|---|
| 08151137 | Feb 2008 | EP | regional |
| Number | Name | Date | Kind |
|---|---|---|---|
| 7319102 | Clark et al. | Jan 2008 | B1 |
| 7968557 | Choi et al. | Jun 2011 | B2 |
| 20080096903 | Chen et al. | Apr 2008 | A1 |
| 20080139588 | Clark et al. | Jun 2008 | A1 |
| 20090163468 | Chen et al. | Jun 2009 | A1 |
| 20090181959 | Rodgers et al. | Jul 2009 | A1 |
| 20090192176 | Zask et al. | Jul 2009 | A1 |
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| 1382339 | Jan 2004 | EP |
| 2006241089 | Sep 2006 | JP |
| 9965908 | Dec 1999 | WO |
| WO 02076985 | Oct 2002 | WO |
| 03074530 | Sep 2003 | WO |
| 2005085253 | Sep 2005 | WO |
| WO 2005080393 | Sep 2005 | WO |
| 2005107760 | Nov 2005 | WO |
| WO 2006045828 | May 2006 | WO |
| 2006074985 | Jul 2006 | WO |
| WO 2006096270 | Sep 2006 | WO |
| 2006122003 | Nov 2006 | WO |
| 2006127406 | Nov 2006 | WO |
| 2007042299 | Apr 2007 | WO |
| WO 2007109362 | Sep 2007 | WO |
| 2007140222 | Dec 2007 | WO |
| WO 2007140222 | Dec 2007 | WO |
| 2008049105 | Apr 2008 | WO |
| 2008157207 | Dec 2008 | WO |
| 2009070524 | Jun 2009 | WO |
| 2009085185 | Jul 2009 | WO |
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| Number | Date | Country | |
|---|---|---|---|
| 20090203688 A1 | Aug 2009 | US |
