Patent Application
20160046610
The invention relates to heteroaryl substituted indazole compounds, a process for their production and the use thereof.
One of the most fundamental characteristics of cancer cells is their ability to sustain chronic proliferation whereas in normal tissues the entry into and progression through the cell divison cycle is tightly controlled to ensure a homeostasis of cell number and maintenance of normal tissue function. Loss of proliferation control was emphasized as one of the six hallmarks of cancer [Hanahan D and Weinberg R A, Cell 100, 57, 2000; Hanahan D and Weinberg R A, Cell 144, 646, 2011].
The eukaryotic cell division cycle (or cell cycle) ensures the duplication of the genome and its distribution to the daughter cells by passing through a coordinated and regulated sequence of events. The cell cycle is divided into four successive phases:
1. The G1 phase represents the time before the DNA replication, in which the cell grows and is sensitive to external stimuli.
2. In the S phase the cell replicates its DNA, and
3. in the G2 phase preparations are made for entry into mitosis.
4. In mitosis (M phase), the duplicated chromosomes get separated supported by a spindle device built from microtubules, and cell division into two daughter cells is completed.
To ensure the extraordinary high fidelity required for an accurate distribution of the chromosomes to the daughter cells, the passage through the cell cycle is strictly regulated and controlled. The enzymes that are necessary for the progression through the cycle must be activated at the correct time and are also turned off again as soon as the corresponding phase is passed. Corresponding control points (“checkpoints”) stop or delay the progression through the cell cycle if DNA damage is detected, or the DNA replication or the creation of the spindle device is not yet completed. The mitotic checkpoint (also known as spindle checkpoint or spindle assembly checkpoint) controls the accurate attachment of mircrotubules of the spindle device to the kinetochors (the attachment site for microtubules) of the duplicated chromosomes. The mitotic checkpoint is active as long as unattached kinetochores are present and generates a wait-signal to give the dividing cell the time to ensure that each kinetochore is attached to a spindle pole, and to correct attachment errors. Thus the mitotic checkpoint prevents a mitotic cell from completing cell division with unattached or erroneously attached chromosomes [Suijkerbuijk S J and Kops G J, Biochem. Biophys. Acta 1786, 24, 2008; Musacchio A and Salmon E D, Nat. Rev. Mol. Cell. Biol. 8, 379, 2007]. Once all kinetochores are attached with the mitotic spindle poles in a correct bipolar (amphitelic) fashion, the checkpoint is satisfied and the cell enters anaphase and proceeds through mitosis.
The mitotic checkpoint is established by a complex network of a number of essential proteins, including members of the MAD (mitotic arrest deficient, MAD 1-3) and Bub (Budding uninhibited by benzimidazole, Bub 1-3) families, Mps1 kinase, cdc20, as well as other components [reviewed in Bolanos-Garcia V M and Blundell T L, Trends Biochem. Sci. 36, 141, 2010], many of these being over-expressed in proliferating cells (e.g. cancer cells) and tissues [Yuan B et al., Clin. Cancer Res. 12, 405, 2006]. The major function of an unsatisfied mitotic checkpoint is to keep the anaphase-promoting complex/cyclosome (APC/C) in an inactive state. As soon as the checkpoint gets satisfied the APC/C ubiquitin-ligase targets cyclin B and securin for proteolytic degradation leading to separation of the paired chromosomes and exit from mitosis.
Inactive mutations of the Ser/Thr kinase Bub1 prevented the delay in progression through mitosis upon treatment of cells of the yeast S. cerevisiae with microtubule-destabilizing drugs, which led to the identification of Bub1 as a mitotic checkpoint protein [Roberts B T et al., Mol. Cell Biol., 14, 8282, 1994]. A number of recent publications provide evidence that Bub1 plays multiple roles during mitosis which, have been reviewed by Elowe [Elowe S, Mol. Cell. Biol. 31, 3085, 2011. In particular, Bub1 is one of the first mitotic checkpoint proteins that binds to the kinetochores of duplicated chromosomes and probably acts as a scaffolding protein to constitute the mitotic checkpoint complex. Furthermore, via phosphorylation of histone H2A, Bub1 localizes the protein shugoshin to the centromeric region of the chromosomes to prevent premature segregation of the paired chromosomes [Kawashima et al. Science 327, 172, 2010]. In addition, together with a Thr-3 phosphorylated Histone H3 the shugoshin protein functions as a binding site for the chromosomal passenger complex which includes the proteins survivin, borealin, INCENP and Aurora B. The chromosomal passenger complex is seen as a tension sensor within the mitotic checkpoint mechanism, which dissolves erroneously formed microtubule-kinetochor attachments such as syntelic (both sister kinetochors are attached to one spindle pole) or merotelic (one kinetochor is attached to two spindle poles) attachments [Watanabe Y, Cold Spring Harb. Symp. Quant. Biol. 75, 419, 2010]. Recent data suggest that the phosphorylation of histone H2A at Thr 121 by Bub1 kinase is sufficient to localize AuroraB kinase to fulfill the attachment error correction checkpoint [Ricke et al. J. Cell Biol. 199, 931-949, 2012].
Incomplete mitotic checkpoint function has been linked with aneuploidy and tumourigenesis [Weaver B A and Cleveland D W, Cancer Res. 67, 10103, 2007; King R W, Biochim Biophys Acta 1786, 4, 2008]. In contrast, complete inhibition of the mitotic checkpoint has been recognised to result in severe chromosome missegregation and induction of apoptosis in tumour cells [Kops G J et al., Nature Rev. Cancer 5, 773, 2005; Schmidt M and Medema R H, Cell Cycle 5, 159, 2006; Schmidt M and Bastians H, Drug Res. Updates 10, 162, 2007]. Thus, mitotic checkpoint abrogation through pharmacological inhibition of components of the mitotic checkpoint, such as Bub1 kinase, represents a new approach for the treatment of proliferative disorders, including solid tumours such as carcinomas, sarcomas, leukaemias and lymphoid malignancies or other disorders, associated with uncontrolled cellular proliferation.
The present invention relates to chemical compounds that inhibit Bub1 kinase.
Established anti-mitotic drugs such as vinca alkaloids, taxanes or epothilones activate the mitotic checkpoint, inducing a mitotic arrest either by stabilising or destabilising microtubule dynamics. This arrest prevents separation of the duplicated chromosomes to form the two daughter cells. Prolonged arrest in mitosis forces a cell either into mitotic exit without cytokinesis (mitotic slippage or adaption) or into mitotic catastrophe leading to cell death [Rieder C L and Maiato H, Dev. Cell 7, 637, 2004]. In contrast, inhibitors of Bub1 prevent the establishment and/or functionality of the mitotic checkpoint, which finally results in severe chromosomal missegregation, induction of apoptosis and cell death.
These findings suggest that Bub1 inhibitors should be of therapeutic value for the treatment of proliferative disorders associated with enhanced uncontrolled proliferative cellular processes such as, for example, cancer, inflammation, arthritis, viral diseases, cardiovascular diseases, or fungal diseases in a warm-blooded animal such as man.
WO 2013/050438, WO 2013/092512, WO 2013/167698 disclose substituted benzylindazoles, substituted benzylpyrazoles and substituted benzylcycloalkylpyrazoles, respectively, which are Bub1 kinase inhibitors.
Due to the fact that especially cancer disease as being expressed by uncontrolled proliferative cellular processes in tissues of different organs of the human- or animal body still is not considered to be a controlled disease in that sufficient drug therapies already exist, there is a strong need to provide further new therapeutically useful drugs, preferably inhibiting new targets and providing new therapeutic options (e.g. drugs with improved pharmacological properties).
Therefore, inhibitors of Bub1 represent valuable compounds that should complement therapeutic options either as single agents or in combination with other drugs.
In accordance with a first aspect, the invention relates to compounds of formula (I),
(i) —NR12R13,
R7 is (a) hydrogen,
R8 is (a) 5-membered heteroaryl,
or an N-oxide, a salt, a tautomer or a stereoisomer of said compound, or a salt of said N-oxide, tautomer or stereoisomer.
In accordance with a variant of the first aspect, the invention relates to compounds of formula (I)
(c) 1-4C-haloalkyl,
or an N-oxide, a salt, a tautomer or a stereoisomer of said compound, or a salt of said N-oxide, tautomer or stereoisomer.
In a second aspect, the invention relates to compounds of formula (I) as described supra,
or an N-oxide, a salt, a tautomer or a stereoisomer of said compound, or a salt of said N-oxide, tautomer or stereoisomer.
In accordance with a variant of the second aspect the invention relates to compounds of formula (I) according to claim 1,
(h) —O-(2-3C-alkylen)-O-(1-3C-alkyl) which is optionally substituted with hydroxy,
or an N-oxide, a salt, a tautomer or a stereoisomer of said compound, or a salt of said N-oxide, tautomer or stereoisomer.
In accordance with a third aspect, the invention relates to compounds of formula (I) as described supra,
or an N-oxide, a salt, a tautomer or a stereoisomer of said compound, or a salt of said N-oxide, tautomer or stereoisomer.
Another aspect of the invention relates to compounds of formula (I) according to claim 1,
or an N-oxide, a salt, a tautomer or a stereoisomer of said compound, or a salt of said N-oxide, tautomer or stereoisomer.
In accordance with a fourth aspect, the invention relates to compounds of formula (I) according to claim 1,
or an N-oxide, a salt, a tautomer or a stereoisomer of said compound, or a salt of said N-oxide, tautomer or stereoisomer.
In a further aspect the invention relates to compounds of formula (I) according to claim 1,
In accordance with a variant of the fifth aspect, the invention relates to compounds of formula (I) according to claim 1,
In another aspect the invention relates to compounds of formula (I) according to claim 1,
wherein said 5-membered heteroaryl or 6-membered heteroaryl or phenyl is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, -(2-3C-alkylen)-O-(1-3C-alkyl), NR12R13,
In accordance with a sixth aspect, the invention relates to compounds of formula (I) according to claim 1,
In a further aspect the invention relates to compounds of formula (I) according to claim 1,
In a further aspect of the invention compounds of formula (I) as described above are selected from the group consisting of:
In one aspect of the invention compounds of formula (I) as described above are selected from the group consisting of:
One aspect of the invention are compounds of formula (I) as described in the examples as characterized by their names in the title as claimed in claim 7 and their structures as well as the subcombinations of all residues specifically disclosed in the compounds of the examples.
Another aspect of the present invention are the intermediates as used for their synthesis.
Another aspect of the invention relates to the use of any of the intermediates described herein for preparing a compound of formula (I) as defined herein or an N-oxide, a salt, a tautomer or a stereoisomer of said compound, or a salt of said N-oxide, tautomer or stereoisomer.
Another aspect of the invention are compounds of formula (I), wherein
R1 is hydrogen, halogen, 1-3C-alkyl.
Yet another aspect of the invention are compounds of formula (I) according to claims 1, 2, 3, 4, 5 or 6, wherein R1 is hydrogen.
A further aspect of the invention are compounds of formula (I), wherein
R2/R3 are independently from each other hydrogen, halogen, cyano, hydroxy 1-6C-haloalkyl, 1-6C-haloalkoxy, 1-6C-alkoxy,
A further aspect of the invention are compounds of formula (I) according to claim 1, wherein R2 and/or R3 are independently from each other hydrogen or halogen, preferably fluorine.
Another aspect of the invention are compounds of formula (I), wherein
R2 and/or R3 is halogen, especially fluorine, chlorine or bromine, preferably fluorine or chlorine, more preferably fluorine.
A further aspect of the invention are compounds of formula (I), wherein
R2 and R3 are different.
Another aspect of the invention are compounds of formula (I), wherein
R4 is independently hydrogen, hydroxy, halogen, cyano, 1-6C-alkyl, 2-6C-alkenyl, 2-6C-alkynyl, 1-6C-haloalkyl, 1-6C-hydroxyalkyl, 1-6C-alkoxy, —O-(2-6alkylen)-O—C(O)-(1-6C-alkyl), 1-6C-haloalkoxy, —C(O)OR9, —C(O)-(1-6C-alkyl), —C(O)NR10R11, 3-7C-cycloalkyl, —S(O)2NH-(3-7C-cycloalkyl), —S(O)2NR10R11.
Another aspect of the invention are compounds of formula (I), wherein
R4 is heteroaryl which optionally is substituted independently one or more times with cyano, 1-4C-alkyl, 1-6C-haloalkyl, 1-6C-haloalkoxy.
Another aspect of the invention are compounds of formula (I), wherein whereby two of R2, R3 (R4)n, when positioned ortho to each other, may form together with the two carbon atoms to which they are attached, a heterocyclic 5-, 6- or 7-membered ring containing 1 or 2 heteroatoms selected from O or N, and optionally containing an additional double bond and/or optionally substituted by an oxo (═O) group and/or an 1-4C-alkyl group.
Another aspect of the invention are compounds of formula (I), wherein
R4 is hydrogen.
Another aspect of the invention are compounds of formula (I), wherein
R4 is hydrogen, halogen, 1-6C-alkyl, 1-6C-alkoxy.
Another aspect of the invention are compounds of formula (I), wherein
R4 is hydrogen, halogen, 1-3C-alkyl, 1-3C-alkoxy.
Another aspect of the invention are compounds of formula (I), wherein
R4 is hydrogen, halogen or 1-6C-alkoxy, preferably hydrogen, fluorine, propyl methoxy or ethoxy.
In another embodiment of the above-mentioned aspects, the invention relates to compounds of formula (I), wherein n is 0 or 1.
Another aspect of the invention are compounds of formula (I), wherein
n is 1.
Another aspect of the invention are compounds of formula (I), wherein
R6 is (a) hydrogen;
Another aspect of the invention are compounds of formula (I), wherein
R6 is hydrogen, hydroxy or methoxy.
Another aspect of the invention are compounds of formula (I), wherein R7 is hydrogen,
Another aspect of the invention are compounds of formula (I), wherein
R8 is a 5-membered heteroaryl group, preferably selected from the group consisting of pyrazolyl, oxazolyl, thiazolyl, triazolyl (1,2,4-triazolyl, 1,3,4-triazolyl or 1,2,3-triazolyl), more preferably 1H-pyrazol-4-yl, 1H-pyrazol-5-yl, 1,2-thiazol-4-yl, 1,2,4-triazol-5-yl, which are optionally substituted with methyl, ethyl, amino, —(CH2)—O—CH2—CH3.
Another aspect of the invention are compounds of formula (I), wherein
R8 is (a) 5-membered heteroaryl, preferably selected from the group consisting of 1H-pyrazol-4-yl, 1H-pyrazol-5-yl, 1,2-thiazol-4-yl, 4H-1,2,4-triazol-3-yl, 1H-1,2,4-triazol-5-yl,
(b) 6-membered heteroaryl selected from
pyridin-2-yl, pyridin-3-yl, pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1,3,5-triazin-2-yl,
(c) phenyl,
Another aspect of the invention are compounds of formula (I), wherein
R8 is (a) 5-membered heteroaryl, preferably selected from the group consisting of 1H-pyrazol-4-yl, 1H-pyrazol-5-yl, 1,2-thiazol-4-yl, 4H-1,2,4-triazol-3-yl, 1H-1,2,4-triazol-5-yl,
(b) 6-membered heteroaryl selected from pyridin-2-yl, pyridin-3-yl, pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1,3,5-triazin-2-yl, 1,2,4-triazin-3-yl, 1,2,4-triazin-5-yl, 1,2,4-triazin-6-yl,
(c) phenyl,
Another aspect of the invention are compounds of formula (I), wherein R8 is
(a) 5-membered heteroaryl, preferably selected from the group consisting of 1H-pyrazol-4-yl, 1H-pyrazol-5-yl, 1,2-thiazol-4-yl, 4H-1,2,4-triazol-3-yl, 1H-1,2,4-triazol-5-yl,
(b) 6-membered heteroaryl selected from
(b1) pyridin-2-yl, pyridin-3-yl, pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1,3,5-triazin-2-yl,
Another aspect of the invention are compounds of formula (I), wherein R8 is
(a) 5-membered heteroaryl, preferably selected from the group consisting of 1H-pyrazol-4-yl, 1H-pyrazol-5-yl, 1,2-thiazol-4-yl, 4H-1,2,4-triazol-3-yl, 1H-1,2,4-triazol-5-yl,
(b) 6-membered heteroaryl selected from
(b1) pyridin-2-yl, pyridin-3-yl, pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1,3,5-triazin-2-yl, 1,2,4-triazin-3-yl, 1,2,4-triazin-5-yl, 1,2,4-triazin-6-yl,
Another aspect of the invention are compounds of formula (I), wherein R8 is
(a) 5-membered heteroaryl, preferably selected from the group consisting of 1H-pyrazol-4-yl, 1H-pyrazol-5-yl, 1,2-thiazol-4-yl, 4H-1,2,4-triazol-3-yl, 1H-1,2,4-triazol-5-yl,
(b) 6-membered heteroaryl selected from
pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1,3,5-triazin-2-yl,
Another aspect of the invention are compounds of formula (I), wherein R8 is
(a) 5-membered heteroaryl, preferably selected from the group consisting of 1H-pyrazol-4-yl, 1H-pyrazol-5-yl, 1,2-thiazol-4-yl, 4H-1,2,4-triazol-3-yl, 1H-1,2,4-triazol-5-yl,
(b) 6-membered heteroaryl selected from
pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1,3,5-triazin-2-yl,
Another aspect of the invention are compounds of formula (I), wherein R8 is
(a) 5-membered heteroaryl, preferably selected from the group consisting of 1H-pyrazol-4-yl, 1H-pyrazol-5-yl, 1,2-thiazol-4-yl, 4H-1,2,4-triazol-3-yl, 1H-1,2,4-triazol-5-yl,
(b) 6-membered heteroaryl selected from
pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, 1,3,5-triazin-2-yl,
Another aspect of the invention are compounds of formula (I), wherein R8 is
(a) 5-membered heteroaryl, preferably selected from the group consisting of 1H-pyrazol-4-yl, 1H-pyrazol-5-yl, 1,2-thiazol-4-yl, 4H-1,2,4-triazol-3-yl, 1H-1,2,4-triazol-5-yl,
(b) 6-membered heteroaryl selected from
pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, 1,3,5-triazin-2-yl,
Another aspect of the invention are compounds of formula (I), wherein R8 is
(a) 5-membered heteroaryl, preferably selected from the group consisting of 1H-pyrazol-4-yl, 1H-pyrazol-5-yl, 1,2-thiazol-4-yl, 4H-1,2,4-triazol-3-yl, 1H-1,2,4-triazol-5-yl,
(b) 6-membered heteroaryl selected from
pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1,3,5-triazin-2-yl,
or phenyl,
wherein said 5-membered heteroaryl or 6-membered heteroaryl or phenyl is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, -(2-3C-alkylen)-O-(1-3C-alkyl), NH2.
Another aspect of the invention are compounds of formula (I), wherein R8 is
(a) 5-membered heteroaryl, preferably selected from the group consisting of 1H-pyrazol-4-yl, 1H-pyrazol-5-yl, 1,2-thiazol-4-yl, 4H-1,2,4-triazol-3-yl, 1H-1,2,4-triazol-5-yl,
(b) 6-membered heteroaryl selected from
pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1,3,5-triazin-2-yl,
or phenyl,
wherein said 5-membered heteroaryl or 6-membered heteroaryl or phenyl is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, —(CH2)—O-(1-3C-alkyl), ethoxymethyl-, -(2-3C-alkylen)-O-(1-3C-alkyl), —C(O)OR9, —C(O)NR10R11, NH2.
Another aspect of the invention are compounds of formula (I), wherein
R8 is a 5-membered heteroaryl group or a 6-membered heteroaryl group containing 2-3 nitrogen atoms which is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, -(2-3C-alkylen)-O-(1-3C-alkyl), —NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is a 5-membered heteroaryl group or a 6-membered heteroaryl group containing 2-3 nitrogen atoms which is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, —(CH2)—O-(1-3C-alkyl), ethoxymethyl-, -(2-3C-alkylen)-O-(1-3C-alkyl), —C(O)NR10R11, —NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is phenyl, which is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, -(2-3C-alkylen)-O-(1-3C-alkyl), NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is phenyl, which is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, —(CH2)—O-(1-3C-alkyl), ethoxymethyl-, -(2-3C-alkylen)-O-(1-3C-alkyl), —C(O)OR9, —C(O)NR11R11, —NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is phenyl, which is optionally substituted independently one or more times with fluorine, hydroxy, —C(O)OR9, —C(O)NR10R11.
Another aspect of the invention are compounds of formula (I), wherein
R8 is phenyl, which is optionally substituted independently one or more times with fluorine, hydroxy.
Another aspect of the invention are compounds of formula (I), wherein
R8 is a 5-membered heteroaryl group which is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, -(2-3C-alkylen)-O-(1-3C-alkyl), NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is a 5-membered heteroaryl group which is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, —(CH2)—O-(1-3C-alkyl), ethoxymethyl-, -(2-3C-alkylen)-O-(1-3C-alkyl), —C(O)NR10R11, —NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is a 5-membered heteroaryl group which is optionally substituted independently one or more times with 1-3C-alkyl, —(CH2)—O-(1-3C-alkyl), ethoxymethyl-, —NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is a 5-membered heteroaryl group containing 1-3 heteroatoms selected from O, S, N, especially a 5-membered heteroaryl group containing 2-3 heteroatoms selected from S or N, atoms which is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, -(2-3C-alkylen)-O-(1-3C-alkyl), NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is a 5-membered heteroaryl group containing 1-3 heteroatoms selected from O, S, N, especially a 5-membered heteroaryl group containing 2-3 heteroatoms selected from S or N, atoms which is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, —(CH2)—O-(1-3C-alkyl), ethoxymethyl-, -(2-3C-alkylen)-O-(1-3C-alkyl), —C(O)NR10R11, —NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is a 5-membered heteroaryl group containing 1-3 heteroatoms selected from O, S, N, especially a 5-membered heteroaryl group containing 2-3 heteroatoms selected from S or N, atoms which is optionally substituted independently one or more times with 1-3C-alkyl, —(CH2)—O-(1-3C-alkyl), ethoxymethyl-, —NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is a 6-membered heteroaryl group containing 2-3 nitrogen atoms which is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, -(2-3C-alkylen)-O-(1-3C-alkyl), NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is a 6-membered heteroaryl group containing 2-3 nitrogen atoms which is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, —(CH2)—O-(1-3C-alkyl), ethoxymethyl-, -(2-3C-alkylen)-O-(1-3C-alkyl), —C(O)OR9, —C(O)NR10R11, —NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is a 6-membered heteroaryl group containing 2-3 nitrogen atoms which is optionally substituted independently one or more times with —C(O)NR10R11.
Another aspect of the invention are compounds of formula (I), wherein
R8 is a 6-membered heteroaryl group consisting of at least two heteroatoms atoms which is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, -(2-3C-alkylen)-O-(1-3C-alkyl), NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is a 6-membered heteroaryl group consisting of at least two heteroatoms atoms which is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, —(CH2)—O-(1-3C-alkyl), ethoxymethyl-, -(2-3C-alkylen)-O-(1-3C-alkyl), —C(O)OR9, —C(O)NR10R11, —NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is pyridin-2-yl, pyridin-3-yl, pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1,3,5-triazin-2-yl, each of which is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, -(2-3C-alkylen)-O-(1-3C-alkyl), NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R8 is pyridin-2-yl, pyridin-3-yl, pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1,3,5-triazin-2-yl, each of which is optionally substituted independently one or more times with fluorine, hydroxy, 1-3C-alkyl, —(CH2)—O-(1-3C-alkyl), ethoxymethyl-, -(2-3C-alkylen)-O-(1-3C-alkyl), —C(O)OR9, —C(O)NR10R11, —NR12R13.
Another aspect of the invention are compounds of formula (I), wherein R8 is a 6-membered heteroaryl selected from pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1,3,5-triazin-2-yl, wherein said 6-membered heteroaryl is optionally substituted independently one or more times with 1-3C-alkyl, -(2-3C-alkylen)-O-(1-3C-alkyl), NR12R13,
Another aspect of the invention are compounds of formula (I), wherein R8 is a 6-membered heteroaryl selected from pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1,3,5-triazin-2-yl, wherein said 6-membered heteroaryl is optionally substituted independently one or more times with 1-3C-alkyl, —(CH2)—O-(1-3C-alkyl), ethoxymethyl-, -(2-3C-alkylen)-O-(1-3C-alkyl), —C(O)NR10R11, NR12R13.
Another aspect of the invention are compounds of formula (I), wherein R8 is a 6-membered heteroaryl selected from pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl and 1,3,5-triazin-2-yl, wherein said 6-membered heteroaryl is optionally substituted independently one or more times with 1-3C-alkyl, -(2-3C-alkylen)-O-(1-3C-alkyl), NR12R13,
Another aspect of the invention are compounds of formula (I), wherein R8 is a 6-membered heteroaryl selected from pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl and 1,3,5-triazin-2-yl, wherein said 6-membered heteroaryl is optionally substituted independently one or more times with 1-3C-alkyl, —(CH2)—O-(1-3C-alkyl), ethoxymethyl-, -(2-3C-alkylen)-O-(1-3C-alkyl), —C(O)NR10R11, NR12R13.
Another aspect of the invention are compounds of formula (I), wherein
R9 is hydrogen.
Still another aspect of the invention are compounds of formula (I), wherein n is 1.
Another aspect of the invention are compounds of formula (I), wherein n is 0 or 1.
Another aspect of the invention are compounds of formula (I), wherein
R12, R13 are independently from each other hydrogen, 1-4C-alkyl, 2-4C-hydroxyalkyl, —C(O)-(1-6C-alkyl), —C(O)-(1-6C-alkylen)-O-(1-6C-alkyl), —CHO, C(O)OR9.
Another aspect of the invention are compounds of formula (I), wherein
R12, R13 are hydrogen.
Another aspect of the invention are compounds of formula (I), wherein
R10/R11 are independently from each other hydrogen, —C(O)-(1-6-alkylen)-O-(-6C-alkyl).
Another aspect of the invention are compounds of formula (I), wherein
R10/R11 are independently from each other hydrogen, 1-4C-alkyl, preferably H and methyl.
Another aspect of the invention are compounds of formula (I), wherein
R10/R11 are hydrogen.
Another aspect of the invention are compounds of formula (I), wherein T is CH,
Another aspect of the invention are compounds of formula (I), wherein T is N,
Another aspect of the invention are compounds of formula (I), wherein V is CH.
Another aspect of the invention are compounds of formula (I), wherein V is N.
Another aspect of the invention are compounds of formula (I), wherein Y is CR6.
Another aspect of the invention are compounds of formula (I), wherein Y is N.
A further aspect of the invention are compounds of formula (I), which are present as their salts.
It is to be understood that the present invention relates to any sub-combination within any embodiment or aspect of the present invention of compounds of general formula (I), supra.
More particularly still, the present invention covers compounds of general formula (I) which are disclosed in the Example section of this text, infra.
In accordance with another aspect, the present invention covers methods of preparing compounds of the present invention, said methods comprising the steps as described in the Experimental Section herein.
Another embodiment of the invention are compounds according to the claims as disclosed in the Claims section wherein the definitions are limited according to the preferred or more preferred definitions as disclosed below or specifically disclosed residues of the exemplified compounds and subcombinations thereof.
Constituents which are optionally substituted as stated herein, may be substituted, unless otherwise noted, one or more times, independently from one another at any possible position. When any variable occurs more than one time in any constituent, each definition is independent. For example, when R1, R2, R3, R4, R6, R7, R8, R9, R10, R11, R12, R13 T, V and/or Y occur more than one time for any compound of formula (I) each definition of R1, R2, R3, R4, R6, R7, R8, R9, R10, R11, R12, R13 T, V and Y is independent.
Should a constituent be composed of more than one part, e.g. —O-(1-6Calkyl)-(3-7C-cycloalkyl), the position of a possible substituent can be at any of these parts at any suitable position. A hyphen at the beginning of the constituent marks the point of attachment to the rest of the molecule. Should a ring be substituted the substitutent could be at any suitable position of the ring, also on a ring nitrogen atom if suitable.
The term “comprising” when used in the specification includes “consisting of”.
If it is referred to “as mentioned above” or “mentioned above” within the description it is referred to any of the disclosures made within the specification in any of the preceding pages.
“suitable” within the sense of the invention means chemically possible to be made by methods within the knowledge of a skilled person.
“1-6C-alkyl” is a straight-chain or branched alkyl group having 1 to 6 carbon atoms. Examples are methyl, ethyl, n propyl, iso-propyl, n butyl, iso-butyl, sec-butyl and tert-butyl, pentyl, hexyl, preferably 1-4 carbon atoms (1-4C-alkyl), more preferably 1-3 carbon atoms (1-3C-alkyl). Other alkyl constituents mentioned herein having another number of carbon atoms shall be defined as mentioned above taking into account the different length of their chain. Those parts of constituents containing an alkyl chain as a bridging moiety between two other parts of the constituent which usually is called an “alkylene” moiety is defined in line with the definition for alkyl above including the preferred length of the chain e.g. methylen, ethylene, n-propylen, iso-propylen, n-butylen, isobutylene, tert-butylen.
“2-6C-Alkenyl” is a straight chain or branched alkenyl radical having 2 to 6 carbon atoms, particularly 2 or 3 carbon atoms (“2-3-C-Alkenyl”). Examples are the but-2-enyl, but-3-enyl (homoallyl), prop-1-enyl, prop-2-enyl (allyl) and the ethenyl (vinyl) radicals.
“2-6C-Alkynyl” is a straight chain or branched alkynyl radical having 2 to 6 carbon atoms, particularly 2 or 3 carbon atoms (“2-3C-Alkynyl”). Examples are the ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, pent-1-ynyl, pent-2-ynyl, pent-3-ynyl, pent-4-ynyl, hex-1-ynyl, hex-2-inyl, hex-3-inyl, hex-4-ynyl, hex-5-ynyl, 1-methylprop-2-ynyl, 2-methylbut-3-ynyl, 1-methylbut-3-ynyl, 1-methylbut-2-ynyl, 3-methylbut-1-ynyl, 1-ethylprop-2-ynyl, 3-methylpent-4-ynyl, 2-methylpent-4-ynyl, 1-methylpent-4-ynyl, 2-methylpent-3-ynyl, 1-methylpent-3-ynyl, 4-methylpent-2-ynyl, 1-methylpent-2-ynyl, 4-methylpent-1-ynyl, 3-methylpent-1-ynyl, 2-ethylbut-3-ynyl, 1-ethylbut-3-ynyl, 1-ethylbut-2-ynyl, 1-propylprop-2-ynyl, 1-isopropylprop-2-ynyl, 2,2-dimethylbut-3-inyl, 1,1-dimethylbut-3-ynyl, 1,1-dimethylbut-2-ynyl, or 3,3-dimethylbut-1-ynyl radicals. Particularly, said alkynyl group is ethynyl, prop-1-ynyl, or prop-2-inyl.
“Halogen” within the meaning of the present invention is iodine, bromine, chlorine or fluorine, preferably “halogen” within the meaning of the present invention is chlorine or fluorine.
“1-6C-Haloalkyl” is a straight-chain or branched alkyl group having 1 to 6 carbon atoms in which at least one hydrogen is substituted by a halogen atom. Examples are chloromethyl or 2-bromoethyl, preferably 1-4 carbon atoms (1-4C-haloalkyl), more preferably 1-3 carbon atoms (1-3C-haloalkyl). For a partially or completely fluorinated C1-C4-alkyl group, the following partially or completely fluorinated groups are considered, for example: fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl, 1,1-difluoroethyl, 1,2-difluoroethyl, 1,1,1-trifluoroethyl, tetrafluoroethyl, and penta-fluoroethyl, whereby difluoromethyl, trifluoromethyl, or 1,1,1-trifluoroethyl are preferred. All possible partially or completely fluorinated 1-6C-alkyl groups are considered to be encompassed by the term 1-6C-haloalkyl.
“1-6C-Hydroxyalkyl” is a straight-chain or branched alkyl group having 1 to 6 carbon atoms in which at least one hydrogen atom is substituted by a hydroxy group, preferably 1-4 carbon atoms (1-4C-hydroxyalkyl), more preferably 1-3 carbon atoms (1-3C-hydroxyalkyl). Examples are hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 1,2-dihydroxyethyl, 3-hydroxypropyl, 2-hydroxypropyl, 2,3-dihydroxypropyl, 3-hydroxy-2-methyl-propyl, 2-hydroxy-2-methyl-propyl, 1-hydroxy-2-methyl-propyl.
“1-6C-Alkoxy” represents radicals, which in addition to the oxygen atom, contain a straight-chain or branched alkyl radical having 1 to 6 carbon atoms, preferably 1-4 carbon atoms (1-4C-alkoxy), more preferably 1-3 carbon atoms (1-3C-alkoxy). Examples which may be mentioned are the hexoxy, pentoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, propoxy, isopropoxy, ethoxy and methoxy radicals, preferred are methoxy, ethoxy, propoxy, isopropoxy. In case the alkoxy group may be substituted those substituents as defined (d1)-(d11) may be situated at any carbon atom of the alkyoxy group being chemically suitable.
“1-6C-Haloalkoxy” represents radicals, which in addition to the oxygen atom, contain a straight-chain or branched alkyl radical having 1 to 6 carbon atoms in which at least one hydrogen is substituted by a halogen atom, preferably 1-4 carbon atoms (1-4C-haloalkoxy), more preferably 1-3 carbon atoms (1-3C-haloalkoxy). Examples are —O—CFH2, —O—CF2H, —O—CF3, —O—CH2—CFH2, —O—CH2—CF2H, —O—CH2—CF3.
“3-7C-Cycloalkyl” stands for cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl, preferably cyclopropyl.
“3-7C-Cycloalkyloxy” represents radicals, which in addition to the oxygen atom, contain a 3-7C-cycloalkyl radical. Examples which may be mentioned are the cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy or cycloheptyloxy radicals.
“3-7C-Heterocyclyl”, or “heterocyclyl” represents a mono- or polycyclic, preferably mono- or bicyclic, more preferably monocyclic, nonaromatic heterocyclic radical containing, 4 to 10, preferably 4 to 7, more preferably 5 to 6 ring atoms, and 1, 2 or 3, preferably 1 or 2, hetero atoms and/or hetero groups independently selected from the series consisting of N, O, S, SO, SO2. The heterocyclyl radicals can be saturated or partially unsaturated and, unless stated otherwise, may be optionally substituted, one or more times, identically or differently, with a substituent selected from: 1-4C-alkyl, 1-4C-haloalkyl, 1-4C-alkoxy, hydroxy, fluorine or (═O) whereby the 1-4C-alkyl may be optionally further substituted with hydroxy and the double bonded oxygen atom leads to a carbonyl group together with the carbon atom of the heterocyclyl ring at any suitable position. Particularly preferred heterocyclic radicals are 4- to 7-membered monocyclic saturated heterocyclyl radicals having up to two hetero atoms from the series consisting of O, N and S, more preferred 5-6-membered heterocyclic radicals. The following may be mentioned by way of example and by preference: oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, azetidinyl, 3-hydroxyazetidinyl, 3-fluoroazetidinyl, 3,3-difluoroazetidinyl, pyrrolidinyl, 3-hydroxypyrrolidinyl, pyrrolinyl, pyrazolidinyl, imidazolidinyl, piperidinyl, 3-hydroxypiperidinyl, 4-hydroxypiperidinyl, 3-fluoropiperidinyl, 3,3-difluoropiperidinyl, 4-fluoropiperidinyl, 4,4-difluoropiperidinyl, piperazinyl, N-methyl-piperazinyl, N-(2-hydroxyethyl)-piperazinyl, morpholinyl, thiomorpholinyl, azepanyl, homopiperazinyl, N-methyl-homopiperazinyl.
“N-heterocyclyl” represents a heterocyclic radical which is connected to the remaining molecule via its nitrogen atom contained in the heterocyclic ring.
The term “heteroaryl” represents a monocyclic 5- or 6-membered aromatic heterocycle or a fused bicyclic aromatice moiety comprising without being restricted thereto, the 5-membered heteroaryl radicals furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl, triazolyl (1,2,4-triazolyl, 1,3,4-triazolyl or 1,2,3-triazolyl), thiadiazolyl (1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,2,3-thiadiazolyl or 1,2,4-thiadiazolyl) and oxadiazolyl (1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,3-oxadiazolyl or 1,2,4-oxadiazolyl), as well as the 6-membered heteroaryl radicals pyridinyl, pyrimidinyl, pyrazinyl and pyridazinyl as well as the fused ring systems such as e.g. phthalidyl-, thiophthalidyl-, indolyl-, isoindolyl-, dihydroindolyl-, dihydroisoindolyl-, indazolyl-, benzothiazolyl-, benzofuranyl-, benzimidazolyl-, benzoxazinonyl-, chinolinyl-, isochinolinyl-, chinazolinyl-, chinoxalinyl-, cinnolinyl-, phthalazinyl-, 1,7- or 1,8-naphthyridinyl-, cumarinyl-, isocumarinyl-, indolizinyl-, isobenzofuranyl-, azaindolyl-, azaisoindolyl-, furanopyridyl-, furanopyrimidinyl-, furanopyrazinyl-, furanopyidazinyl-, preferred fused ring system is indazolyl. Preferred 5- or 6-membered heteroaryl radicals are furanyl, thienyl, pyrrolyl, thiazolyl, oxazolyl, thiadiazolyl, oxadiazolyl, pyridinyl, pyrimidinyl, pyrazinyl or pyridazinyl. More preferred 5- or 6-membered heteroaryl radicals are furan-2-yl, thien-2-yl, pyrrol-2-yl, thiazolyl, oxazolyl, 1,3,4-thiadiazolyl, 1,3,4-oxadiazolyl, pyridin-2-yl, pyridin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrazin-2-yl or pyridazin-3-yl.
The term “5-membered heteroaryl” represents a monocyclic 5-membered aromatic heterocyclic ring comprising without being restricted thereto the radicals furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl, triazolyl (1,2,4-triazolyl, 1,3,4-triazolyl or 1,2,3-triazolyl), thiadiazolyl (1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,2,3-thiadiazolyl or 1,2,4-thiadiazolyl) and oxadiazolyl (1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,3-oxadiazolyl or 1,2,4-oxadiazolyl.
In case of doubts regarding the name used in the description or claims the structural formula as disclosed in the experimental section shall be decisive.
In general and unless otherwise mentioned, the heteroarylic or heteroarylenic radicals include all the possible isomeric forms thereof, e.g. the positional isomers thereof. Thus, for some illustrative non-restricting example, the term pyridinyl or pyridinylene includes pyridin-2-yl, pyridin-2-ylene, pyridin-3-yl, pyridin-3-ylene, pyridin-4-yl and pyridin-4-ylene; or the term thienyl or thienylene includes thien-2-yl, thien-2-ylene, thien-3-yl and thien-3-ylene.
The heteroarylic, heteroarylenic, or heterocyclic groups mentioned herein may be substituted by their given substituents or parent molecular groups, unless otherwise noted, at any possible position, such as e.g. at any substitutable ring carbon or ring nitrogen atom. Analogously it is being understood that it is possible for any heteroaryl or heterocyclyl group to be attached to the rest of the molecule via any suitable atom if chemically suitable. Unless otherwise noted, any heteroatom of a heteroarylic or heteroarylenic ring with unsatisfied valences mentioned herein is assumed to have the hydrogen atom(s) to satisfy the valences. Unless otherwise noted, rings containing quaternizable amino- or imino-type ring nitrogen atoms (—N═) may be preferably not quaternized on these amino- or imino-type ring nitrogen atoms by the mentioned substituents or parent molecular groups.
The NR12R13 group includes, for example, NH2, N(H)CH3, N(CH3)2, N(H)CH2CH3 and N(CH3)CH2CH3. In the case of —NR12R13, when R12 and R13 together with the nitrogen atom to which they are attached form a 4-6-membered heterocyclic ring optionally containing one further heteroatom selected from the group consisting of O, S or N, the term “heterocyclic ring” is defined above. Especially preferred is morpholinyl.
The C(O)NR10R11 group includes, for example, C(O)NH2, C(O)N(H)CH3, C(O)N(CH3)2, C(O)N(H)CH2CH3, C(O)N(CH3)CH2CH3 or C(O)N(CH2CH3)2. If R10 or R11 are not hydrogen, they may be substituted by hydroxy,
In the case of —NR12R13, when R12 and R13 together with the nitrogen atom to which they are attached form a 4-6-membered heterocyclic ring, the term “heterocyclic ring” is defined above and can be used analogously for C(O)NR10R11.
The C(O)OR9 group includes for example C(O)OH, C(O)OCH3, C(O)OC2H5, C(O)C3H7, C(O)CH(CH3)2, C(O)OC4H9, C(O)OC5H11, C(O)OC6H13; for C(O)O(1-6Calkyl), the alkyl part may be straight or branched and may be substituted.
In the context of the properties of the compounds of the present invention the term “pharmacokinetic profile” means one single parameter or a combination thereof including permeability, bioavailability, exposure, and pharmacodynamic parameters such as duration, or magnitude of pharmacological effect, as measured in a suitable experiment. Compounds with improved pharmacokinetic profiles can, for example, be used in lower doses to achieve the same effect, may achieve a longer duration of action, or a may achieve a combination of both effects.
Salts of the compounds according to the invention include all inorganic and organic acid addition salts and salts with bases, especially all pharmaceutically acceptable inorganic and organic acid addition salts and salts with bases, particularly all pharmaceutically acceptable inorganic and organic acid addition salts and salts with bases customarily used in pharmacy.
One aspect of the invention are salts of the compounds according to the invention including all inorganic and organic acid addition salts, especially all pharmaceutically acceptable inorganic and organic acid addition salts, particularly all pharmaceutically acceptable inorganic and organic acid addition salts customarily used in pharmacy. Another aspect of the invention are the salts with di- and tricarboxylic acids.
Examples of acid addition salts include, but are not limited to, hydrochlorides, hydrobromides, phosphates, nitrates, sulfates, salts of sulfamic acid, formates, acetates, propionates, citrates, D-gluconates, benzoates, 2-(4-hydroxybenzoyl)-benzoates, butyrates, salicylates, sulfosalicylates, lactates, maleates, laurates, malates, fumarates, succinates, oxalates, malonates, pyruvates, acetoacetates, tartarates, stearates, benzensulfonates, toluenesulfonates, methanesulfonates, trifluoromethansulfonates, 3-hydroxy-2-naphthoates, benzenesulfonates, naphthalinedisulfonates and trifluoroacetates.
Examples of salts with bases include, but are not limited to, lithium, sodium, potassium, calcium, aluminum, magnesium, titanium, meglumine, ammonium, salts optionally derived from NH3 or organic amines having from 1 to 16C-atoms such as e.g. ethylamine, diethylamine, triethylamine, ethyldiisopropylamine, monoethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, dimethylaminoethanol, procaine, dibenzylamine, N-methylmorpholine, arginine, lysine, ethylendiamine, N-methylpiperindine and and guanidinium salts.
The salts include water-insoluble and, particularly, water-soluble salts.
In the present text, in particular in the Experimental Section, for the synthesis of intermediates and of examples of the present invention, when a compound is mentioned as a salt form with the corresponding base or acid, the exact stoichiometric composition of said salt form, as obtained by the respective preparation and/or purification process, is, in most cases, unknown.
Unless specified otherwise, suffixes to chemical names or structural formulae such as “hydrochloride”, “trifluoroacetate”, “sodium salt”, or “x HCl”, “x CF3COOH”, “x Na+”, for example, are to be understood as not a stoichiometric specification, but solely as a salt form.
This applies analogously to cases in which synthesis intermediates or example compounds or salts thereof have been obtained, by the preparation and/or purification processes described, as solvates, such as hydrates with (if defined) unknown stoichiometric composition.
According to the person skilled in the art the compounds of formula (I) according to this invention as well as their salts may contain, e.g. when isolated in crystalline form, varying amounts of solvents. Included within the scope of the invention are therefore all solvates and in particular all hydrates of the compounds of formula (I) according to this invention as well as all solvates and in particular all hydrates of the salts of the compounds of formula (I) according to this invention.
The term “combination” in the present invention is used as known to persons skilled in the art and may be present as a fixed combination, a non-fixed combination or kit-of-parts.
A “fixed combination” in the present invention is used as known to persons skilled in the art and is defined as a combination wherein the said first active ingredient and the said second active ingredient are present together in one unit dosage or in a single entity. One example of a “fixed combination” is a pharmaceutical composition wherein the said first active ingredient and the said second active ingredient are present in admixture for simultaneous administration, such as in a formulation. Another example of a “fixed combination” is a pharmaceutical combination wherein the said first active ingredient and the said second active ingredient are present in one unit without being in admixture.
A non-fixed combination or “kit-of-parts” in the present invention is used as known to persons skilled in the art and is defined as a combination wherein the said first active ingredient and the said second active ingredient are present in more than one unit. One example of a non-fixed combination or kit-of-parts is a combination wherein the said first active ingredient and the said second active ingredient are present separately. The components of the non-fixed combination or kit-of-parts may be administered separately, sequentially, simultaneously, concurrently or chronologically staggered. Any such combination of a compound of formula (I) of the present invention with an anti-cancer agent as defined below is an embodiment of the invention.
The term “(chemotherapeutic) anti-cancer agents”, includes but is not limited to 131I-chTNT, abarelix, abiraterone, aclarubicin, aldesleukin, alemtuzumab, alitretinoin, altretamine, aminoglutethimide, amrubicin, amsacrine, anastrozole, arglabin, arsenic trioxide, asparaginase, azacitidine, basiliximab, BAY 80-6946, BAY 1000394, belotecan, bendamustine, bevacizumab, bexarotene, bicalutamide, bisantrene, bleomycin, bortezomib, buserelin, busulfan, cabazitaxel, calcium folinate, calcium levofolinate, capecitabine, carboplatin, carmofur, carmustine, catumaxomab, celecoxib, celmoleukin, cetuximab, chlorambucil, chlormadinone, chlormethine, cisplatin, cladribine, clodronic acid, clofarabine, copanlisib, crisantaspase, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, darbepoetin alfa, dasatinib, daunorubicin, decitabine, degarelix, denileukin diftitox, denosumab, deslorelin, dibrospidium chloride, docetaxel, doxifluridine, doxorubicin, doxorubicin+estrone, eculizumab, edrecolomab, elliptinium acetate, eltrombopag, endostatin, enocitabine, epirubicin, epitiostanol, epoetin alfa, epoetin beta, eptaplatin, eribulin, erlotinib, estradiol, estramustine, etoposide, everolimus, exemestane, fadrozole, filgrastim, fludarabine, fluorouracil, flutamide, formestane, fotemustine, fulvestrant, gallium nitrate, ganirelix, gefitinib, gemcitabine, gemtuzumab, glutoxim, goserelin, histamine dihydrochloride, histrelin, hydroxycarbamide, 1-125 seeds, ibandronic acid, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib, imiquimod, improsulfan, interferon alfa, interferon beta, interferon gamma, ipilimumab, irinotecan, ixabepilone, lanreotide, lapatinib, lenalidomide, lenograstim, lentinan, letrozole, leuprorelin, levamisole, lisuride, lobaplatin, lomustine, lonidamine, masoprocol, medroxyprogesterone, megestrol, melphalan, mepitiostane, mercaptopurine, methotrexate, methoxsalen, Methyl aminolevulinate, methyltestosterone, mifamurtide, miltefosine, miriplatin, mitobronitol, mitoguazone, mitolactol, mitomycin, mitotane, mitoxantrone, nedaplatin, nelarabine, nilotinib, nilutamide, nimotuzumab, nimustine, nitracrine, ofatumumab, omeprazole, oprelvekin, oxaliplatin, p53 gene therapy, paclitaxel, palifermin, palladium-103 seed, pamidronic acid, panitumumab, pazopanib, pegaspargase, PEG-epoetin beta (methoxy PEG-epoetin beta), pegfilgrastim, peginterferon alfa-2b, pemetrexed, pentazocine, pentostatin, peplomycin, perfosfamide, picibanil, pirarubicin, plerixafor, plicamycin, poliglusam, polyestradiol phosphate, polysaccharide-K, porfimer sodium, pralatrexate, prednimustine, procarbazine, quinagolide, radium-223 chloride, raloxifene, raltitrexed, ranimustine, razoxane, refametinib, regorafenib, risedronic acid, rituximab, romidepsin, romiplostim, sargramostim, sipuleucel-T, sizofiran, sobuzoxane, sodium glycididazole, sorafenib, streptozocin, sunitinib, talaporfin, tamibarotene, tamoxifen, tasonermin, teceleukin, tegafur, tegafur+gimeracil+oteracil, temoporfin, temozolomide, temsirolimus, teniposide, testosterone, tetrofosmin, thalidomide, thiotepa, thymalfasin, tioguanine, tocilizumab, topotecan, toremifene, tositumomab, trabectedin, trastuzumab, treosulfan, tretinoin, trilostane, triptorelin, trofosfamide, tryptophan, ubenimex, valrubicin, vandetanib, vapreotide, vemurafenib, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, vorinostat, vorozole, yttrium-90 glass microspheres, zinostatin, zinostatin stimalamer, zoledronic acid, zorubicin.
The compounds of the present invention may exist as tautomers. For example, any compound of the present invention which contains a pyrazole moiety as a heteroaryl group for example can exist as a 1H tautomer, or a 2H tautomer, or even a mixture in any amount of the two tautomers, or a triazole moiety for example can exist as a 1H tautomer, a 2H tautomer, or a 4H tautomer, or even a mixture in any amount of said 1H, 2H and 4H tautomers. Other examples of such compounds are hydroxypyridines and hydroxypyrimidines which can exist as tautomeric forms:
Another embodiment of the invention are all possible tautomers of the compounds of the present invention as single tautomers, or as any mixture of said tautomers, in any ratio.
The compounds of the invention may, depending on their structure, exist in different stereoisomeric forms. These forms include configurational isomers or optionally conformational isomers (enantiomers and/or diastereoisomers including those of atropisomers). The present invention therefore includes enantiomers, diastereoisomers as well as mixtures thereof. From those mixtures of enantiomers and/or disastereoisomers pure stereoisomeric forms can be isolated with methods known in the art, preferably methods of chromatography, especially high pressure liquid chromatography (HPLC) using achiral or chiral phase. The invention further includes all mixtures of the stereoisomers mentioned above independent of the ratio, including the racemates.
Furthermore, the present invention includes all possible crystalline forms, or polymorphs, of the compounds of the present invention, either as single polymorphs, or as a mixture of more than one polymorph, in any ratio.
Furthermore, derivatives of the compounds of formula (I) and the salts thereof which are converted into a compound of formula (I) or a salt thereof in a biological system (bioprecursors or pro-drugs) are covered by the invention. Said biological system is e.g. a mammalian organism, particularly a human subject. The bioprecursor is, for example, converted into the compound of formula (I) or a salt thereof by metabolic processes.
The invention also includes all suitable isotopic variations of a compound of the invention. An isotopic variation of a compound of the invention is defined as one in which at least one atom is replaced by an atom having the same atomic number but an atomic mass different from the atomic mass usually or predominantly found in nature. Examples of isotopes that can be incorporated into a compound of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulphur, fluorine, chlorine, bromine and iodine, such as 2H (deuterium), 3H (tritium), 11C, 13C, 14C, 15N, 17O, 18O, 32P, 33P, 33S, 34S, 35S, 36S, 18F, 36Cl, 82Br, 123I, 124I, 129I and 131I, respectively. Certain isotopic variations of a compound of the invention, for example, those in which one or more radioactive isotopes such as 3H or 14C are incorporated, are useful in drug and/or substrate tissue distribution studies. Tritiated and carbon-14, i.e., 14C, isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with isotopes such as deuterium may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements and hence may be preferred in some circumstances. Isotopic variations of a compound of the invention can generally be prepared by conventional procedures known by a person skilled in the art such as by the illustrative methods or by the preparations described in the examples hereafter using appropriate isotopic variations of suitable reagents.
It has now been found, and this constitutes the basis of the present invention, that said compounds of the present invention have surprising and advantageous properties.
In particular, said compounds of the present invention have surprisingly been found to effectively inhibit Bub1 kinase and may therefore be used for the treatment or prophylaxis of diseases of uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune responses, or inappropriate cellular inflammatory responses or diseases which are accompanied with uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune responses, or inappropriate cellular inflammatory responses, particularly in which the uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune responses, or inappropriate cellular inflammatory responses is mediated by Bub1 kinase, such as, for example, haematological tumours, solid tumours, and/or metastases thereof, e.g. leukaemias and myelodysplastic syndrome, malignant lymphomas, head and neck tumours including brain tumours and brain metastases, tumours of the thorax including non-small cell and small cell lung tumours, gastrointestinal tumours, endocrine tumours, mammary and other gynaecological tumours, urological tumours including renal, bladder and prostate tumours, skin tumours, and sarcomas, and/or metastases thereof.
The intermediates used for the synthesis of the compounds of claims 1-7 as described below, as well as their use for the synthesis of the compounds of claims 1-7, are one further aspect of the present invention. Preferred intermediates are the Intermediate Examples as disclosed below.
General Procedures
The compounds according to the invention can be prepared according to the following schemes 1 through 6.
The schemes and procedures described below illustrate synthetic routes to the compounds of general formula (I) of the invention and are not intended to be limiting. It is obvious to the person skilled in the art that the order of transformations as exemplified in the Schemes can be modified in various ways. The order of transformations exemplified in the Schemes is therefore not intended to be limiting. In addition, interconversion of any of the substituents, R1, R2, R3, R4, R6, R7 or R8 can be achieved before and/or after the exemplified transformations. These modifications can be such as the introduction of protecting groups, cleavage of protecting groups, reduction or oxidation of functional groups, halogenation, metallation, substitution or other reactions known to the person skilled in the art. These transformations include those which introduce a functionality which allows for further interconversion of substituents. Appropriate protecting groups and their introduction and cleavage are well-known to the person skilled in the art (see for example T. W. Greene and P. G. M. Wuts in Protective Groups in Organic Synthesis, 3rd edition, Wiley 1999). Specific examples are described in the subsequent paragraphs.
One route for the preparation of compounds of general formula (Ia) is described in Scheme 1. In instances where this route is not feasible, scheme 2 can be applied.
Scheme 1 Route for the preparation of compounds of general formula (Ia), wherein R1, R2, R3, R4, R6, R8, T and n have the meaning as given for general formula (I), supra. In addition, interconversion of any of the substituents, R1, R2, R3, R4, R6 or R8 can be achieved before and/or after the exemplified transformations. These modifications can be such as the introduction of protecting groups, cleavage of protecting groups, reduction or oxidation of functional groups, halogenation, metallation, substitution or other reactions known to the person skilled in the art. These transformations include those which introduce a functionality which allows for further interconversion of substituents. Appropriate protecting groups and their introduction and cleavage are well-known to the person skilled in the art (see for example T. W. Greene and P. G. M. Wuts in Protective Groups in Organic Synthesis, 3rd edition, Wiley 1999). Specific examples are described in the subsequent paragraphs.
Compounds A, B, and C are either commercially available or can be prepared according to procedures available from the public domain, as understandable to the person skilled in the art. Specific examples are described in the subsequent paragraphs. X represents a leaving group such as for example a CI, Br or I, or X stands for an aryl sulfonate such as for example p-toluene sulfonate, or for an alkyl sulfonate such as for example methane sulfonate or trifluoromethane sulfonate. X′ represents F, Cl, Br, I, boronic acid or a boronic acid ester, such as for example 4,4,5,5-tetramethyl-2-phenyl-1,3,2-dioxaborolane (boronic acid pinacole ester).
A suitably substituted carbonitrile (A) can be reacted with a suitably substituted benzyl halide or benzyl sulfonate of general formula (B), such as, for example, a benzyl bromide, in a suitable solvent system, such as, for example, N,N-dimethylformamide, in the presence of a suitable base, such as, for example, cesium carbonate at temperatures ranging from −78° C. to room temperature, preferably the reaction is carried out at room temperature, to furnish general formula (1-1).
Intermediates of general formula (1-1) can be converted to intermediates of general formula (1-2) by reaction with a suitable alcoholate, such as, for example sodium methanolate, in a suitable solvent system, such as, for example, the corresponding alcohol, e.g. methanol, at a temperature between room temperature and the boiling point of the respective solvent, preferably the reaction is carried out at room temperature, and subsequent treatment with a suitable source of ammonium, such as for example, ammonium chloride in the presence of a suitable acid, such as for example acetic acid in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 50° C.
Intermediates of general formula (1-2) are reacted with a suitably substituted 3,3-bis(dimethylamino)propanenitrile of the general formula (1-3), such as, for example 3,3-bis(dimethylamino)-2-methoxypropanenitrile, in the presence of a suitable base, such as, for example piperidine, in a suitable solvent system, such as, for example, 3-methylbutan-1-ol, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 100° C., to furnish intermediates of general formula (1-4).
Intermediates of general formula (1-4) can be reacted with a suitable 4-haloaromatic or heteroaromatic system of the general formula (C), such as, for example 4-chloropyrimidine, in the presence of a suitable base, such as, for example sodium 2-methylpropan-2-olate or potassium carbonate. Optionally, a suitable palladium catalyst, such as for example (1E,4E)-1,5-diphenylpenta-1,4-dien-3-one-palladium or palladium (II) acetate, and a suitable ligand, such as for example 1′-binaphthalene-2,2′-diylbis(diphenylphosphane) or (9,9-dimethyl-9H-xanthene-4,5-diyl)bis(diphenylphosphine), can be added. The reaction is carried out in a suitable solvent system, such as, for example, N,N-dimethylformamide, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 105° C. to furnish compounds of general formula (Ia). Alternatively, the following palladium catalysts can be used: Allylpalladium chloride dimer, Dichlorobis(benzonitrile)palladium (II), Palladium (II) chloride, Tetrakis(triphenylphosphine)palladium (0), Tris(dibenzylideneacetone)-dipalladium (0), optionally with addition of the following ligands: racemic-2,2′-Bis(diphenylphosphino)-1,1′-binaphthyl, rac-BINAP, 1,1′-Bis(diphenyl-phosphino)ferrocene, Bis(2-diphenylphosphinophenyl)ether, Di-t-butylmethylphos-phonium tetrafluoroborate, 2-(Di-t-butylphosphino)biphenyl, Tri-t-butylphospho-nium tetrafluoroborate, Tri-2-furylphosphine, or Tris(2,4-di-t-butylphenyl)phosphite, Tri-o-tolylphosphine.
Alternatively, intermediates of general formula (1-4) can be reacted with a suitable boronic acid or boronic acid pinacole ester of general formula (C), such as, for example pyridin-3-ylboronic acid, in the presence of a suitable base, such as, for example triethylamine, a suitable activating agent such as for example N,N-dimethylpyridin-4-amine and a suitable copper salt, such as for example copper (II) acetate, in a suitable solvent system, such as, for example, trichloromethane, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at room temperature to furnish compounds of general formula (Ia).
Alternatively, intermediates of general formula (1-4) can be reacted with suitable compound of the general formula (C), such as, for example 4-bromo-pyrimidin hydrochloride, in the presence of a suitable base, such as, for example potassium carbonate, in a suitable solvent system, such as, for example, dimethyl formamide, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 100° C. to furnish 20 compounds of general formula (Ia).
An alternative route for the preparation of compounds of general formula (Ia) is described in Scheme 1a. In instances where this route is not feasible, scheme 2 can be applied.
Scheme 1a Route for the preparation of compounds of general formula (Ia), wherein R1, R2, R3, R4, R6, R8 T and n have the meaning as given for general formula (I), supra. In addition, interconversion of any of the substituents, R1, R2, R3, R4, R6 or R8 can be achieved before and/or after the exemplified transformations. These modifications can be such as the introduction of protecting groups, cleavage of protecting groups, reduction or oxidation of functional groups, halogenation, metallation, substitution or other reactions known to the person skilled in the art. These transformations include those which introduce a functionality which allows for further interconversion of substituents. Appropriate protecting groups and their introduction and cleavage are well-known to the person skilled in the art (see for example T. W. Greene and P. G. M. Wuts in Protective Groups in Organic Synthesis, 3rd edition, Wiley 1999). Specific examples are described in the subsequent paragraphs.
Compounds E, B, and C are either commercially available or can be prepared according to procedures available from the public domain, as understandable to the person skilled in the art. Specific examples are described in the subsequent paragraphs. X represents a leaving group such as for example a CI, Br or I, or X stands for an aryl sulfonate such as for example p-toluene sulfonate, or for an alkyl sulfonate such as for example methane sulfonate or trifluoromethane sulfonate. X′ represents F, Cl, Br, I, boronic acid or a boronic acid ester, such as for example 4,4,5,5-tetramethyl-2-phenyl-1,3,2-dioxaborolane (boronic acid pinacole ester). R′″ represents an alkyl-group, such as for example methyl or ethyl.
A suitably substituted ester (E) can be reacted with a suitably substituted benzyl halide or benzyl sulfonate of general formula (B), such as, for example, a benzyl bromide, in a suitable solvent system, such as, for example, N,N-dimethylformamide, in the presence of a suitable base, such as, for example, cesium carbonate at temperatures ranging from −78° C. to room temperature, preferably the reaction is carried out at room temperature, to furnish general formula (1-6).
Intermediates of general formula (1-6) can be converted to intermediates of general formula (1-2) by reaction with methylchloroaluminium amide generated in situ, in a suitable solvent system, such as, for example, toluene, at a temperature between 0° C. and the boiling point of the respective solvent, preferably the reaction is carried out at 80° C., and subsequent treatment with methanol in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 0° C.
The following intermediates and compounds can be prepared using the synthetic methods described in context of Scheme 1 Compounds of general formula (I) can also be synthesised according to the procedure depicted in Scheme 2.
Scheme 2 Alternative route for the preparation of compounds of general formula (I), wherein R1, R2, R3, R4, R7, R8, T, V, Y and n have the meaning as given for general formula (I), supra. R′ is for example alkyl or benzyl, preferably methyl or ethyl. In addition, interconversion of any of the substituents, R1, R2, R3, R4, R7 or R8 can be achieved before and/or after the exemplified transformations. These modifications can be such as the introduction of protecting groups, cleavage of protecting groups, reduction or oxidation of functional groups, halogenation, metallation, substitution or other reactions known to the person skilled in the art.
These transformations include those which introduce a functionality which allows for fur-ther interconversion of substituents. Appropriate protecting groups and their intro-duction and cleavage are well-known to the person skilled in the art (see for ex-ample T. W. Greene and P. G. M. Wuts in Protective Groups in Organic Synthesis, 3rd edition, Wiley 1999). Further specific examples are described in the subsequent paragraphs.
Compounds of the formula (Ib) can be prepared using the synthetic methods described in context of Scheme 1; the introduction of R7 different from hydrogen may be accomplished inter alia by the methods described in Scheme 5. Compounds B are either commercially available or can be prepared according to procedures available from the public domain, as understandable to the person skilled in the art as referred to below scheme 1 above.
Compounds of general formula (Ib) are converted to intermediates of general formula (1-5) by treatment with a suitable acid system, such as, for example a mixture of trifluoroacetic acid and trifluoromethanesulfonic acid, in a suitable solvent, such as, for example, dichloroethan, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at room temperature.
Intermediates of general formula (1-5) can be reacted with a suitably substituted benzyl halide or benzyl sulfonate of general formula (B), such as, for example, a benzyl bromide, in a suitable solvent system, such as, for example, tetrahydrofuran, in the presence of a suitable base, such as, for example, sodium hydride in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at room temperature, to furnish compounds of general formula (I). Said reaction can also result in double conversion of intermediate (1-5) if R7 is hydrogen, giving rise to compounds formed alongside the target compounds, in which R7 is a benzylic group identical with the benzylic moiety attached to the indazole nitrogen.
Compounds of general formula (Ie) and (Id) can be synthesised from compounds of general formula (Ic), according to the procedure depicted in Scheme 3.
Scheme 3 Process for the preparation of compounds of general formula (Ie) via de-methylation of compounds of general formula (Ic) and subsequent etherification to furnish compounds of general formula (Ie), wherein R1, R2, R3, R4, R7, R8, T, V and n have the meaning as given for general formula (I), supra. In addition, interconversion of any of the substituents, R1, R2, R3, R4, R7 or R8 can be achieved before and/or after the exemplified transformations. These modifications can be such as the introduction of protecting groups, cleavage of protecting groups, reduction or oxidation of functional groups, halogenation, metallation, substitution or other reactions known to the person skilled in the art. These transformations include those which introduce a functionality which allows for further interconversion of substituents. Appropriate protecting groups and their introduction and cleavage are well-known to the person skilled in the art (see for example T. W. Greene and P. G. M. Wuts in Protective Groups in Organic Synthesis, 3rd edition, Wiley 1999).
Compounds of the formula (Ic) can be prepared using the synthetic methods described in context of Scheme 1; the introduction of R7 different from hydrogen may be accomplished inter alia by the methods described in Scheme 5.
Compounds of general formula D are commercially available, wherein X represents leaving a group such as for example a CI, Br or I, or X stands for an aryl sulfonate such as for example p-toluene sulfonate, or for an alkyl sulfonate such as for example methane sulfonate or trifluoromethane sulfonate (triflate group). R″=1-6C-alkyl (independently one or more times optionally substituted with hydroxy, C(O)OR9, C(O)NR10R11, NR12R13, —S-(1-6C-alkyl), —S(O)-(1-6C-alkyl), —S(O)2-(1-6C-alkyl), S(O)2NR10R11, heterocyclyl (which itself is optionally substituted with C(O)OR9 or oxo (═O)), heteroaryl (which itself is optionally substituted one or more times with cyano, 1-4C-alkyl, 1-6C-haloalkyl, 1-6C-haloalkoxy, C(O)OR9, C(O)NR10R11, -(2-6C-alkyl)-O-1-6C-alkyl)), 3-7C-cycloalkyl, 1-6C-haloalkyl, or
whereby the * is the point of attachment.
Compounds of general formula (Ic) are converted to compounds of general formula (Id) by treatment with a suitable demethylating agent, such as for example benzenethiol, in a suitable solvent, such as, for example, 1-methylpyrrolidin-2-one, in the presence of a suitable base, such as, for example potassium carbonate, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 190° C.
Compounds of general formula (Id) are then reacted with a compound of general formula (D) as mentioned above, in a suitable solvent, such as, for example, N,N-dimethylformamide, in the presence of a suitable base, such as, for example, potassium carbonate in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at room temperature, to furnish compounds of general formula (Ie).
Compounds of general formula (If′) which is a compound of formula (If) wherein R7=hydrogen, can be converted into compounds of general formula (Ig and Ih) according to the procedure depicted in Scheme 5.
Compounds of general formula (Ij) can be synthesised from compounds of general formula F and G, according to the procedure depicted in Scheme 4.
Scheme 4 Process for the preparation of compounds of general formula (Ij) wherein R1, R2, R3, R4, R8, T, V, Y and n have the meaning as given for general formula (I), supra. In addition, interconversion of any of the substituents, R1, R2, R3, R4, or R8 can be achieved before and/or after the exemplified transformations. These modifications can be such as the introduction of protecting groups, cleavage of protecting groups, reduction or oxidation of functional groups, halogenation, metallation, substitution or other reactions known to the person skilled in the art. These transformations include those which introduce a functionality which allows for further interconversion of substituents. Appropriate protecting groups and their introduction and cleavage are well-known to the person skilled in the art (see for example T. W. Greene and P. G. M. Wuts in Protective Groups in Organic Synthesis, 3rd edition, Wiley 1999).
Compounds B, F, G, H and J are either commercially available or can be prepared according to procedures available from the public domain, as understandable to the person skilled in the art. X″ represents a leaving group such as for example a CI, Br or I. Specific examples are described in the subsequent paragraphs. X represents leaving group such as for example a CI, Br or I, or X stands for an aryl sulfonate such as for example p-toluene sulfonate, or for an alkyl sulfonate such as for example methane sulfonate or trifluoromethane sulfonate (triflate group). X′″ represents leaving group such as for example a CI, Br, I or a boronic acid or boronic acid pinacole ester.
A suitably substituted indazolehalogenide (F) can be reacted with a suitably substituted benzyl halide or benzyl sulfonate of general formula (B), such as, for example, a benzyl bromide, in a suitable solvent system, such as, for example, N,N-dimethylformamide, in the presence of a suitable base, such as, for example, cesium carbonate at temperatures ranging from −78° C. to room temperature, preferably the reaction is carried out at room temperature, to furnish general formula (1-7).
Alternatively a suitably substituted 1,2-dihydro-3H-indazol-3-one (G) can be reacted with a suitably substituted benzyl halide or benzyl sulfonate of general formula (B), such as, for example, a benzyl bromide, in a suitable solvent system, such as, for example, N,N-dimethylformamide, in the presence of a suitable base, such as, for example, potassium carbonate at temperatures ranging from −78° C. to room temperature, preferably the reaction is carried out at room temperature, to furnish general formula (1-8).
Intermediates of general formula (1-8) can be converted to intermediates of general formula (1-7) by reaction with a suitable sulfonilation agent, such as, for example trifluoromethynesulfonic anhydride, in a suitable solvent system, such as, for example, dichloromethane, in the presence of a suitable base, such as, for example, pyridine at temperatures ranging from −78° C. to the boiling point of the respective solvent, preferably the reaction is carried out at room temperature, to furnish general formula (1-7).
Intermediates of general formula (1-7) can be converted to intermediates of general formula (1-9) by reaction with a suitable boronic acid or boronic acid pinacole ester of general formula (H), wherein X′″ is a suitable boronic acid or boronic acid pinacole ester, such as, for example 4-chloro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine, in the presence of a suitable base, such as, for example potassium carbonate, in the presence of a suitable catalyst, such as, for example (1,1,-bis(diphenylphosphino)ferrocene)-dichloropalladium(II) and a suitable copper salt, such as for example copper (I) bromide, in a suitable solvent system, such as, for example, N,N-dimethylformamide, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 100° C. to furnish compounds of general formula (1-9).
Alternatively Intermediates of general formula (1-7) can be converted to intermediates of general formula (1-9) by transforming general formula (1-7) in situ into a stannyl compound by reaction with a suitable stannylation reagent, such as, for example hexamthylditin, in the presence of a suitable catalyst, such as, for example tetrakis(triphenylphosphin)palladium (0), in a suitable solvent system, such as, for example, dioxane, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 100° C. This stannyl compound can be converted to intermediates of general formula (1-9) by reaction with a suitable bis-halo-heteroaryl-compound (H), wherein X′″ is halogene, such as, for example 2-bromo-4-chloropyrimidine, in the presence of a suitable catalyst, such as, for example tetrakis(triphenyl-phosphin)palladium (0), in a suitable solvent system, such as, for example, toluene, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 110° C.
Intermediates of general formula (1-9) can be reacted with a suitable aminoaromatic or heteroaromatic system of the general formula (J), such as, for example pyrimidin-4-amine, in the presence of a suitable base, such as, for example cesium carbonate. Optionally, a suitable palladium catalyst, such as for example palladium (II) acetate, and a suitable ligand, such as for example 1′-binaphthalene-2,2′-diylbis(diphenylphosphane) or (9,9-dimethyl-9H-xanthene-4,5-diyl)bis(diphenylphosphine), can be added. The reaction is carried out in a suitable solvent system, such as, for example, dioxane, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 105° C. to furnish compounds of general formula (Ij). Alternatively, the following palladium catalysts can be used:
Allylpalladium chloride dimer, Dichlorobis(benzonitrile)palladium (II), Palladium (II) chloride, Tetrakis(triphenylphosphine)palladium (0), Tris(dibenzylideneacetone)-dipalladium (0), optionally with addition of the following ligands: racemic-2,2′-Bis(diphenylphosphino)-1,1′-binaphthyl, rac-BINAP, 1,1′-Bis(diphenyl-phosphino)ferrocene, Bis(2-diphenylphosphinophenyl)ether, Di-t-butylmethylphos-phonium tetrafluoroborate, 2-(Di-t-butylphosphino)biphenyl, Tri-t-butylphospho-nium tetrafluoroborate, Tri-2-furylphosphine, or Tris(2,4-di-t-butylphenyl)phosphite, Tri-o-tolylphosphine.
Alternatively, intermediates of general formula (1-9) can be reacted with a compound of general formula (J), such as, for example 1-ethyl-1H-1,2,4-triazol-5-amine, in a suitable solvent system, such as, for example, 1-methyl-2-pyrrolidone, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 200° C. to furnish compounds of general formula (Ij).
Compounds of general formula (Ih) can be synthesised from compounds of general formula (If) and (Ig), according to the procedure depicted in Scheme 4.
Scheme 5. Process for the transformation of compounds of general formula (If) into compounds of general formula (Ig) and (Ih), wherein R1, R2, R3, R4, R7, R8, T, V, Y and n have the meaning as given for general formula (I), supra. In addition, interconversion of any of the substituents, R1, R2, R3, R4, R7a, R7b or R8 can be achieved before and/or after the exemplified transformations. These modifications can be such as the introduction of protecting groups, cleavage of protecting groups, reduction or oxidation of functional groups, halogenation, metallation, substitution or other reactions known to the person skilled in the art. These transformations include those which introduce a functionality which allows for further interconversion of substituents. Appropriate protecting groups and their introduction and cleavage are well-known to the person skilled in the art (see for example T. W. Greene and P. G. M. Wuts in Protective Groups in Organic Synthesis, 3rd edition, Wiley 1999).
R7a represents 1-4C-alkyl, independently one or more times optionally substituted with heteroaryl, halogen, hydroxy, or R7a stands for
whereby the * is the point of attachment, or R7a represents benzyl, whereby the phenyl ring is optionally substituted one or more times with halogen, 1-4Calkyl, 1-4C-haloalkyl, 1-4C-alkoxy, 1-4C-haloalkoxy, cyano, C(O)OR9. X is as defined below scheme 1, supra, or for example represents 1,3,2-dioxathiolane 2-oxide.
R7b represents an acyl moiety, such as —C(O)-(1-6C-alkyl), —C(O)-(1-6C-alkylen)-O-(1-6C-alkyl), —C(O)-(1-6C-alkylen)-O-(2-6C-alkylen)-O-(1-6C-alkyl), —C(O)-heterocyclyl and Z represents a halogen, hydroxy or —O—R7b.
Compounds of general formula (If) are converted into compounds of general formula (Ig) by reaction with a suitable haloalkyl or dioxathiolane 2-oxide, such as, for example 1,3,2-dioxathiolane 2-oxide, in a suitable solvent system, such as, for example, N,N-dimethyl formamide, in the presence of a suitable base, such as, for example cesium carbonate, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 60° C.
Compounds of general formula (If) are converted into compounds of general formula (Ih) by reaction with a suitable carboxylic acid derivative, such as for example a carboxylic acid halogenide e.g. carboxylic acid chloride, or a carboxylic acid anhydride, in a suitable solvent, such as, for example, dichloromethane, in the presence of a suitable base, such as, for example N,N-diethylethanamine, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at room temperature.
Compounds of general formula (Ij) can be synthesised from compounds of general formula 1-7, according to the procedure depicted in Scheme 6.
Scheme 6 Process for the preparation of compounds of general formula (Ij) wherein R1, R2, R3, R4, R8, T, V, Y and n have the meaning as given for general formula (I), supra. In addition, interconversion of any of the substituents, R1, R2, R3, R4, or R8 can be achieved before and/or after the exemplified transformations. These modifications can be such as the introduction of protecting groups, cleavage of protecting groups, reduction or oxidation of functional groups, halogenation, metallation, substitution or other reactions known to the person skilled in the art. These transformations include those which introduce a functionality which allows for further interconversion of substituents. Appropriate protecting groups and their introduction and cleavage are well-known to the person skilled in the art (see for example T. W. Greene and P. G. M. Wuts in Protective Groups in Organic Synthesis, 3rd edition, Wiley 1999).
Compounds of the formula (1-7) can be prepared using the synthetic methods described in context of Scheme 4.
X represents leaving group such as for example a CI, Br or I, or X stands for an aryl sulfonate such as for example p-toluene sulfonate, or for an alkyl sulfonate such as for example methane sulfonate or trifluoromethane sulfonate (triflate group).
Compound K is either commercially available or can be prepared according to procedures available from the public domain, as understandable to the person skilled in the art.
Intermediates of general formula (1-7) can be converted to intermediates of general formula (1-10) by reaction with a suitable boronic acid or boronic acid pinacole ester of general formula (K), where X′″ is a boronic acid or boronic acid pinacole ester), such as, for example 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-4-amine, in the presence of a suitable base, such as, for example potassium carbonate, in the presence of a suitable catalyst, such as, for example (1,1,-bis(diphenylphosphino)ferrocene)-dichloropalladium(II) and a suitable copper salt, such as for example copper (I) bromide, in a suitable solvent system, such as, for example, N,N-dimethylformamide, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 100° C. to furnish compounds of general formula (1-10).
Alternatively Intermediates of general formula (1-7) can be converted to intermediates of general formula (1-10) by reaction with a heteroaryl-halogenide, such as, for example 6-chloropyrimidin-4-amine, in the presence of a suitable catalyst, such as, for example Bis(triphenylphosphin)palladium(II)chlorid, in the presence of a suitable stannylation comounds, such as, for example hexabutylditin, in a suitable solvent system, such as, for example, dioxane, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 100° C. to furnish compounds of general formula (1-10).
Intermediates of general formula (1-10) can be reacted with a suitable aromatic or heteroaromatic compound with a leaving group of the general formula (J), such as, for example 4-chloropyrimidine, in the presence of a suitable base, such as, for example cesium carbonate. Optionally, a suitable palladium catalyst, such as for example palladium (II) acetate, and a suitable ligand, such as for example 1′-binaphthalene-2,2′-diylbis(diphenylphosphane) or (9,9-dimethyl-9H-xanthene-4,5-diyl)bis(diphenylphosphine), can be added. The reaction is carried out in a suitable solvent system, such as, for example, N,N-dimethylformamide, in a temperature range from room temperature to the boiling point of the respective solvent, preferably the reaction is carried out at 105° C. to furnish compounds of general formula (Ij). Alternatively, the following palladium catalysts can be used:
Allylpalladium chloride dimer, Dichlorobis(benzonitrile)palladium (II), Palladium (II) chloride, Tetrakis(triphenylphosphine)palladium (0), Tris(dibenzylideneacetone)-dipalladium (0), optionally with addition of the following ligands:
racemic-2,2′-Bis(diphenylphosphino)-1,1′-binaphthyl, rac-BINAP, 1,1′-Bis(diphenyl-phosphino)ferrocene, Bis(2-diphenylphosphinophenyl)ether, Di-t-butylmethylphos-phonium tetrafluoroborate, 2-(Di-t-butylphosphino)biphenyl, Tri-t-butylphospho-nium tetrafluoroborate, Tri-2-furylphosphine, or Tris(2,4-di-t-butylphenyl)phosphite, Tri-o-tolylphosphine.
One preferred aspect of the invention is the process for the preparation of the compounds of claims 1-7 according to the Examples.
It is known to the person skilled in the art that, if there are a number of reactive centers on a starting or intermediate compound, it may be necessary to block one or more reactive centers temporarily by protective groups in order to allow a reaction to proceed specifically at the desired reaction center. A detailed description for the use of a large number of proven protective groups is found, for example, in T. W. Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, 1999, 3rd Ed., or in P. Kocienski, Protecting Groups, Thieme Medical Publishers, 2000.
The compounds according to the invention are isolated and purified in a manner known per se, e.g. by distilling off the solvent in vacuo and recrystallizing the residue obtained from a suitable solvent or subjecting it to one of the customary purification methods, such as chromatography on a suitable support material.
Furthermore, reverse phase preparative HPLC of compounds of the present invention which possess a sufficiently basic or acidic functionality, may result in the formation of a salt, such as, in the case of a compound of the present invention which is sufficiently basic, a trifluoroacetate or formate salt for example, or, in the case of a compound of the present invention which is sufficiently acidic, an ammonium salt for example. Salts of this type can either be transformed into its free base or free acid form, respectively, by various methods known to the person skilled in the art, or be used as salts in subsequent biological assays. Additionally, the drying process during the isolation of compounds of the present invention may not fully remove traces of cosolvents, especially such as formic acid or trifluoroacetic acid, to give solvates or inclusion complexes. The person skilled in the art will recognise which solvates or inclusion complexes are acceptable to be used in subsequent biological assays. It is to be understood that the specific form (e.g. salt, free base, solvate, inclusion complex) of a compound of the present invention as isolated as described herein is not necessarily the only form in which said compound can be applied to a biological assay in order to quantify the specific biological activity.
Salts of the compounds of formula (I) according to the invention can be obtained by dissolving the free compound in a suitable solvent (for example a ketone such as acetone, methylethylketone or methylisobutylketone, an ether such as diethyl ether, tetrahydrofuran or dioxane, a chlorinated hydrocarbon such as methylene chloride or chloroform, or a low molecular weight aliphatic alcohol such as methanol, ethanol or isopropanol) which contains the desired acid or base, or to which the desired acid or base is then added. The acid or base can be employed in salt preparation, depending on whether a mono- or polybasic acid or base is concerned and depending on which salt is desired, in an equimolar quantitative ratio or one differing therefrom. The salts are obtained by filtering, reprecipitating, precipitating with a non-solvent for the salt or by evaporating the solvent. Salts obtained can be converted into the free compounds which, in turn, can be converted into salts. In this manner, pharmaceutically unacceptable salts, which can be obtained, for example, as process products in the manufacturing on an industrial scale, can be converted into pharmaceutically acceptable salts by processes known to the person skilled in the art. Especially preferred are hydrochlorides and the process used in the example section.
Pure diastereomers and pure enantiomers of the compounds and salts according to the invention can be obtained e.g. by asymmetric synthesis, by using chiral starting compounds in synthesis and by splitting up enantiomeric and diasteriomeric mixtures obtained in synthesis.
Enantiomeric and diastereomeric mixtures can be split up into the pure enantiomers and pure diastereomers by methods known to a person skilled in the art. Preferably, diastereomeric mixtures are separated by crystallization, in particular fractional crystallization, or chromatography. Enantiomeric mixtures can be separated e.g. by forming diastereomers with a chiral auxiliary agent, resolving the diastereomers obtained and removing the chiral auxiliary agent. As chiral auxiliary agents, for example, chiral acids can be used to separate enantiomeric bases such as e.g. mandelic acid and chiral bases can be used to separate enantiomeric acids via formation of diastereomeric salts. Furthermore, diastereomeric derivatives such as diastereomeric esters can be formed from enantiomeric mixtures of alcohols or enantiomeric mixtures of acids, respectively, using chiral acids or chiral alcohols, respectively, as chiral auxiliary agents. Additionally, diastereomeric complexes or diastereomeric clathrates may be used for separating enantiomeric mixtures. Alternatively, enantiomeric mixtures can be split up using chiral separating columns in chromatography. Another suitable method for the isolation of enantiomers is the enzymatic separation.
One preferred aspect of the invention is the process for the preparation of the compounds of claims 1-7 according to the examples as well as the intermediates used for their preparation.
Optionally, compounds of the formula (I) can be converted into their salts, or, optionally, salts of the compounds of the formula (I) can be converted into the free compounds. Corresponding processes are customary for the skilled person.
Optionally, compounds of the formula (I) can be converted into their N-oxides. The N-oxide may also be introduced by way of an intermediate. N-oxides may be prepared by treating an appropriate precursor with an oxidizing agent, such as meta-chloroperbenzoic acid, in an appropriate solvent, such as dichloromethane, at suitable temperatures, such as from 0° C. to 40° C., whereby room temperature is generally preferred. Further corresponding processes for forming N-oxides are customary for the skilled person.
Commercial Utility
As mentioned supra, the compounds of the present invention have surprisingly been found to effectively inhibit Bub1 finally resulting in cell death e.g. apoptosis and may therefore be used for the treatment or prophylaxis of diseases of uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune responses, or inappropriate cellular inflammatory responses, or diseases which are accompanied with uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune responses, or inappropriate cellular inflammatory responses, particularly in which the uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune responses, or inappropriate cellular inflammatory responses is mediated by Bub1, such as, for example, benign and malignant neoplasia, more specifically haematological tumours, solid tumours, and/or metastases thereof, e.g. leukaemias and myelodysplastic syndrome, malignant lymphomas, head and neck tumours including brain tumours and brain metastases, tumours of the thorax including non-small cell and small cell lung tumours, gastrointestinal tumours, endocrine tumours, mammary and other gynaecological tumours, urological tumours including renal, bladder and prostate tumours, skin tumours, and sarcomas, and/or metastases thereof, especially haematological tumours, solid tumours, and/or metastases of breast, bladder, bone, brain, central and peripheral nervous system, cervix, colon, endocrine glands (e.g. thyroid and adrenal cortex), endocrine tumours, endometrium, esophagus, gastrointestinal tumours, germ cells, kidney, liver, lung, larynx and hypopharynx, mesothelioma, ovary, pancreas, prostate, rectum, renal, small intestine, soft tissue, stomach, skin, testis, ureter, vagina and vulva as well as malignant neoplasias including primary tumors in said organs and corresponding secondary tumors in distant organs (“tumor metastases”). Haematological tumors can e.g be exemplified by aggressive and indolent forms of leukemia and lymphoma, namely non-Hodgkins disease, chronic and acute myeloid leukemia (CML/AML), acute lymphoblastic leukemia (ALL), Hodgkins disease, multiple myeloma and T-cell lymphoma. Also included are myelodysplastic syndrome, plasma cell neoplasia, paraneoplastic syndromes, and cancers of unknown primary site as well as AIDS related malignancies.
A further aspect of the invention is the use of the compounds according to formula (I) for the treatment of cer-vical-, breast-, non-small cell lung-, prostate-, colon- and melanoma tumors and/or metastases thereof, especially preferred for the treatment thereof as well as a method of treatment of cervical-, breast-, non-small cell lung-, prostate-, colon- and melanoma tumors and/or metastases thereof comprising administering an effective amount of a compound of formula (I).
One aspect of the invention is the use of the compounds according to formula (I) for the treatment of cervix tumors as well as a method of treatment of cervix tumors comprising administering an effective amount of a compound of formula (I).
In accordance with an aspect of the present invention therefore the invention relates to a compound of general formula I, or an N-oxide, a salt, a tautomer or a stereoisomer of said compound, or a salt of said N-oxide, tautomer or stereoisomer particularly a pharmaceutically acceptable salt thereof, or a mixture of same, as described and defined herein, for use in the treatment or prophylaxis of a disease, especially for use in the treatment of a disease.
Another particular aspect of the present invention is therefore the use of a compound of general formula I, described supra, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, particularly a pharmaceutically acceptable salt thereof, or a mixture of same, for the prophylaxis or treatment of hyperproliferative disorders or disorders responsive to induction of cell death i.e apoptosis.
The term “inappropriate” within the context of the present invention, in particular in the context of “inappropriate cellular immune responses, or inappropriate cellular inflammatory responses”, as used herein, is to be understood as preferably meaning a response which is less than, or greater than normal, and which is associated with, responsible for, or results in, the pathology of said diseases.
Preferably, the use is in the treatment or prophylaxis of diseases, especially the treatment, wherein the diseases are haematological tumours, solid tumours and/or metastases thereof.
Another aspect is the use of a compound of formula (I) is for the treatment of cervical-, breast-, non-small cell lung-, prostate-, colon- and melanoma tumors and/or metastases thereof, especially preferred for the treatment thereof. A preferred aspect is the use of a compound of formula (I) for the prophylaxis and/or treatment of cervical tumors especially preferred for the treatment thereof.
Another aspect of the present invention is the use of a compound of formula (I) or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, particularly a pharmaceutically acceptable salt thereof, or a mixture of same, as described herein, in the manufacture of a medicament for the treatment or prophylaxis of a disease, wherein such disease is a hyperproliferative disorder or a disorder responsive to induction of cell death e.g.apoptosis. In an embodiment the disease is a haematological tumour, a solid tumour and/or metastases thereof. In another embodiment the disease is cervical-, breast-, non-small cell lung-, prostate-, colon- and melanoma tumor and/or metastases thereof, in a preferred aspect the disease is cervical tumor.
Method of Treating Hyper-Proliferative Disorders
The present invention relates to a method for using the compounds of the present invention and compositions thereof, to treat mammalian hyper-proliferative disorders. Compounds can be utilized to inhibit, block, reduce, decrease, etc., cell proliferation and/or cell division, and/or produce cell death e.g. apoptosis. This method comprises administering to a mammal in need thereof, including a human, an amount of a compound of this invention, or a pharmaceutically acceptable salt, isomer, polymorph, metabolite, hydrate, solvate or ester thereof; etc. which is effective to treat the disorder. Hyper-proliferative disorders include but are not limited, e.g., psoriasis, keloids, and other hyperplasias affecting the skin, benign prostate hyperplasia (BPH), solid tumours, such as cancers of the breast, respiratory tract, brain, reproductive organs, digestive tract, urinary tract, eye, liver, skin, head and neck, thyroid, parathyroid and their distant metastases. Those disorders also include lymphomas, sarcomas, and leukaemias.
Examples of breast cancer include, but are not limited to invasive ductal carcinoma, invasive lobular carcinoma, ductal carcinoma in situ, and lobular carcinoma in situ.
Examples of cancers of the respiratory tract include, but are not limited to small-cell and non-small-cell lung carcinoma, as well as bronchial adenoma and pleuropulmonary blastoma.
Examples of brain cancers include, but are not limited to brain stem and hypophtalmic glioma, cerebellar and cerebral astrocytoma, medulloblastoma, ependymoma, as well as neuroectodermal and pineal tumour.
Tumours of the male reproductive organs include, but are not limited to prostate and testicular cancer. Tumours of the female reproductive organs include, but are not limited to endometrial, cervical, ovarian, vaginal, and vulvar cancer, as well as sarcoma of the uterus.
Tumours of the digestive tract include, but are not limited to anal, colon, colorectal, oesophageal, gallbladder, gastric, pancreatic, rectal, small-intestine, and salivary gland cancers.
Tumours of the urinary tract include, but are not limited to bladder, penile, kidney, renal pelvis, ureter, urethral and human papillary renal cancers.
Eye cancers include, but are not limited to intraocular melanoma and retinoblastoma.
Examples of liver cancers include, but are not limited to hepatocellular carcinoma (liver cell carcinomas with or without fibrolamellar variant), cholangiocarcinoma (intrahepatic bile duct carcinoma), and mixed hepatocellular cholangiocarcinoma.
Skin cancers include, but are not limited to squamous cell carcinoma, Kaposi's sarcoma, malignant melanoma, Merkel cell skin cancer, and non-melanoma skin cancer.
Head-and-neck cancers include, but are not limited to laryngeal, hypopharyngeal, nasopharyngeal, oropharyngeal cancer, lip and oral cavity cancer and squamous cell. Lymphomas include, but are not limited to AIDS-related lymphoma, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, Burkitt lymphoma, Hodgkin's disease, and lymphoma of the central nervous system.
Sarcomas include, but are not limited to sarcoma of the soft tissue, osteosarcoma, malignant fibrous histiocytoma, lymphosarcoma, and rhabdomyosarcoma.
Leukemias include, but are not limited to acute myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, and hairy cell leukemia.
These disorders have been well characterized in humans, but also exist with a similar etiology in other mammals, and can be treated by administering pharmaceutical compositions of the present invention.
The term “treating” or “treatment” as stated throughout this document is used conventionally, e.g., the management or care of a subject for the purpose of combating, alleviating, reducing, relieving, improving the condition of, etc., of a disease or disorder, such as a carcinoma.
Methods of Treating Kinase Disorders
The present invention also provides methods for the treatment of disorders associated with aberrant mitogen extracellular kinase activity, including, but not limited to stroke, heart failure, hepatomegaly, cardiomegaly, diabetes, Alzheimer's disease, cystic fibrosis, symptoms of xenograft rejections, septic shock or asthma. Effective amounts of compounds of the present invention can be used to treat such disorders, including those diseases (e.g., cancer) mentioned in the Background section above. Nonetheless, such cancers and other diseases can be treated with compounds of the present invention, regardless of the mechanism of action and/or the relationship between the kinase and the disorder.
The phrase “aberrant kinase activity” or “aberrant tyrosine kinase activity,” includes any abnormal expression or activity of the gene encoding the kinase or of the polypeptide it encodes. Examples of such aberrant activity, include, but are not limited to, over-expression of the gene or polypeptide; gene amplification; mutations which produce constitutively-active or hyperactive kinase activity; gene mutations, deletions, substitutions, additions, etc.
The present invention also provides for methods of inhibiting a kinase activity, especially of mitogen extracellular kinase, comprising administering an effective amount of a compound of the present invention, including salts, polymorphs, metabolites, hydrates, solvates, prodrugs (e.g.: esters) thereof, and diastereoisomeric forms thereof. Kinase activity can be inhibited in cells (e.g., in vitro), or in the cells of a mammalian subject, especially a human patient in need of treatment.
Methods of Treating Angiogenic Disorders
The present invention also provides methods of treating disorders and diseases associated with excessive and/or abnormal angiogenesis.
Inappropriate and ectopic expression of angiogenesis can be deleterious to an organism. A number of pathological conditions are associated with the growth of extraneous blood vessels. These include, e.g., diabetic retinopathy, ischemic retinal-vein occlusion, and retinopathy of prematurity [Aiello et al. New Engl. J. Med. 1994, 331, 1480; Peer et al. Lab. Invest. 1995, 72, 638], age-related macular degeneration [AMD; see, Lopez et al. Invest. Opththalmol. Vis. Sci. 1996, 37, 855], neovascular glaucoma, psoriasis, retrolental fibroplasias, angiofibroma, inflammation, rheumatoid arthritis (RA), restenosis, in-stent restenosis, vascular graft restenosis, etc. In addition, the increased blood supply associated with cancerous and neoplastic tissue, encourages growth, leading to rapid tumour enlargement and metastasis. Moreover, the growth of new blood and lymph vessels in a tumour provides an escape route for renegade cells, encouraging metastasis and the consequence spread of the cancer. Thus, compounds of the present invention can be utilized to treat and/or prevent any of the aforementioned angiogenesis disorders, e.g., by inhibiting and/or reducing blood vessel formation; by inhibiting, blocking, reducing, decreasing, etc. endothelial cell proliferation or other types involved in angiogenesis, as well as causing cell death e.g. apoptosis of such cell types.
Preferably, the diseases of said method are haematological tumours, solid tumour and/or metastases thereof.
The compounds of the present invention can be used in particular in therapy and prevention i.e. prophylaxis, especially in therapy of tumour growth and metastases, especially in solid tumours of all indications and stages with or without pre-treatment of the tumour growth.
Pharmaceutical Compositions of the Compounds of the Invention
This invention also relates to pharmaceutical compositions containing one or more compounds of the present invention. These compositions can be utilised to achieve the desired pharmacological effect by administration to a patient in need thereof. A patient, for the purpose of this invention, is a mammal, including a human, in need of treatment for the particular condition or disease.
Therefore, the present invention includes pharmaceutical compositions that are comprised of a pharmaceutically acceptable carrier or auxiliary and a pharmaceutically effective amount of a compound, or salt thereof, of the present invention.
Another aspect of the invention is a pharmaceutical composition comprising a pharmaceutically effective amount of a compound of formula (I) and a pharmaceutically acceptable auxiliary for the treatment of a disease mentioned supra, especially for the treatment of haematological tumours, solid tumours and/or metastases thereof.
A pharmaceutically acceptable carrier or auxiliary is preferably a carrier that is non-toxic and innocuous to a patient at concentrations consistent with effective activity of the active ingredient so that any side effects ascribable to the carrier do not vitiate the beneficial effects of the active ingredient. Carriers and auxiliaries are all kinds of additives assisting to the composition to be suitable for administration.
A pharmaceutically effective amount of compound is preferably that amount which produces a result or exerts the intended influence on the particular condition being treated.
The compounds of the present invention can be administered with pharmaceutically-acceptable carriers or auxiliaries well known in the art using any effective conventional dosage unit forms, including immediate, slow and timed release preparations, orally, parenterally, topically, nasally, ophthalmically, optically, sublingually, rectally, vaginally, and the like.
For oral administration, the compounds can be formulated into solid or liquid preparations such as capsules, pills, tablets, troches, lozenges, melts, powders, solutions, suspensions, or emulsions, and may be prepared according to methods known to the art for the manufacture of pharmaceutical compositions. The solid unit dosage forms can be a capsule that can be of the ordinary hard- or soft-shelled gelatine type containing auxiliaries, for example, surfactants, lubricants, and inert fillers such as lactose, sucrose, calcium phosphate, and corn starch.
In another embodiment, the compounds of this invention may be tableted with conventional tablet bases such as lactose, sucrose and cornstarch in combination with binders such as acacia, corn starch or gelatine, disintegrating agents intended to assist the break-up and dissolution of the tablet following administration such as potato starch, alginic acid, corn starch, and guar gum, gum tragacanth, acacia, lubricants intended to improve the flow of tablet granulation and to prevent the adhesion of tablet material to the surfaces of the tablet dies and punches, for example talc, stearic acid, or magnesium, calcium or zinc stearate, dyes, colouring agents, and flavouring agents such as peppermint, oil of wintergreen, or cherry flavouring, intended to enhance the aesthetic qualities of the tablets and make them more acceptable to the patient. Suitable excipients for use in oral liquid dosage forms include dicalcium phosphate and diluents such as water and alcohols, for example, ethanol, benzyl alcohol, and polyethylene alcohols, either with or without the addition of a pharmaceutically acceptable surfactant, suspending agent or emulsifying agent. Various other materials may be present as coatings or to otherwise modify the physical form of the dosage unit. For instance tablets, pills or capsules may be coated with shellac, sugar or both.
Dispersible powders and granules are suitable for the preparation of an aqueous suspension. They provide the active ingredient in admixture with a dispersing or wetting agent, a suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients, for example those sweetening, flavouring and colouring agents described above, may also be present.
The pharmaceutical compositions of this invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil such as liquid paraffin or a mixture of vegetable oils. Suitable emulsifying agents may be (1) naturally occurring gums such as gum acacia and gum tragacanth, (2) naturally occurring phosphatides such as soy bean and lecithin, (3) esters or partial esters derived form fatty acids and hexitol anhydrides, for example, sorbitan monooleate, (4) condensation products of said partial esters with ethylene oxide, for example, polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening and flavouring agents.
Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil such as, for example, arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent such as, for example, beeswax, hard paraffin, or cetyl alcohol. The suspensions may also contain one or more preservatives, for example, ethyl or n-propyl p-hydroxybenzoate; one or more colouring agents; one or more flavouring agents; and one or more sweetening agents such as sucrose or saccharin.
Syrups and elixirs may be formulated with sweetening agents such as, for example, glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, and preservative, such as methyl and propyl parabens and flavouring and colouring agents.
The compounds of this invention may also be administered parenterally, that is, subcutaneously, intravenously, intraocularly, intrasynovially, intramuscularly, or interperitoneally, as injectable dosages of the compound in preferably a physiologically acceptable diluent with a pharmaceutical carrier which can be a sterile liquid or mixture of liquids such as water, saline, aqueous dextrose and related sugar solutions, an alcohol such as ethanol, isopropanol, or hexadecyl alcohol, glycols such as propylene glycol or polyethylene glycol, glycerol ketals such as 2,2-dimethyl-1,1-dioxolane-4-methanol, ethers such as poly(ethylene glycol) 400, an oil, a fatty acid, a fatty acid ester or, a fatty acid glyceride, or an acetylated fatty acid glyceride, with or without the addition of a pharmaceutically acceptable surfactant such as a soap or a detergent, suspending agent such as pectin, carbomers, methycellulose, hydroxypropylmethylcellulose, or carboxymethylcellulose, or emulsifying agent and other pharmaceutical adjuvants.
Illustrative of oils which can be used in the parenteral formulations of this invention are those of petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, sesame oil, cottonseed oil, corn oil, olive oil, petrolatum and mineral oil. Suitable fatty acids include oleic acid, stearic acid, isostearic acid and myristic acid. Suitable fatty acid esters are, for example, ethyl oleate and isopropyl myristate. Suitable soaps include fatty acid alkali metal, ammonium, and triethanolamine salts and suitable detergents include cationic detergents, for example dimethyl dialkyl ammonium halides, alkyl pyridinium halides, and alkylamine acetates; anionic detergents, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates; non-ionic detergents, for example, fatty amine oxides, fatty acid alkanolamides, and poly(oxyethylene-oxypropylene)s or ethylene oxide or propylene oxide copolymers; and amphoteric detergents, for example, alkyl-beta-aminopropionates, and 2-alkylimidazoline quarternary ammonium salts, as well as mixtures.
The parenteral compositions of this invention will typically contain from about 0.5% to about 25% by weight of the active ingredient in solution. Preservatives and buffers may also be used advantageously. In order to minimise or eliminate irritation at the site of injection, such compositions may contain a non-ionic surfactant having a hydrophile-lipophile balance (HLB) preferably of from about 12 to about 17. The quantity of surfactant in such formulation preferably ranges from about 5% to about 15% by weight. The surfactant can be a single component having the above HLB or can be a mixture of two or more components having the desired HLB.
Illustrative of surfactants used in parenteral formulations are the class of polyethylene sorbitan fatty acid esters, for example, sorbitan monooleate and the high molecular weight adducts of ethylene oxide with a hydrophobic base, formed by the condensation of propylene oxide with propylene glycol.
The pharmaceutical compositions may be in the form of sterile injectable aqueous suspensions. Such suspensions may be formulated according to known methods using suitable dispersing or wetting agents and suspending agents such as, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents which may be a naturally occurring phosphatide such as lecithin, a condensation product of an alkylene oxide with a fatty acid, for example, polyoxyethylene stearate, a condensation product of ethylene oxide with a long chain aliphatic alcohol, for example, heptadeca-ethyleneoxycetanol, a condensation product of ethylene oxide with a partial ester derived form a fatty acid and a hexitol such as polyoxyethylene sorbitol monooleate, or a condensation product of an ethylene oxide with a partial ester derived from a fatty acid and a hexitol anhydride, for example polyoxyethylene sorbitan monooleate.
The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent. Diluents and solvents that may be employed are, for example, water, Ringer's solution, isotonic sodium chloride solutions and isotonic glucose solutions. In addition, sterile fixed oils are conventionally employed as solvents or suspending media. For this purpose, any bland, fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid can be used in the preparation of injectables.
A composition of the invention may also be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritation excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials are, for example, cocoa butter and polyethylene glycol.
Controlled release formulations for parenteral administration include liposomal, polymeric microsphere and polymeric gel formulations that are known in the art.
It may be desirable or necessary to introduce the pharmaceutical composition to the patient via a mechanical delivery device. The construction and use of mechanical delivery devices for the delivery of pharmaceutical agents is well known in the art. Direct techniques for administration, for example, administering a drug directly to the brain usually involve placement of a drug delivery catheter into the patient's ventricular system to bypass the blood-brain barrier. One such implantable delivery system, used for the transport of agents to specific anatomical regions of the body, is described in U.S. Pat. No. 5,011,472, issued Apr. 30, 1991.
The compositions of the invention can also contain other conventional pharmaceutically acceptable compounding ingredients, generally referred to as carriers or diluents, as necessary or desired. Conventional procedures for preparing such compositions in appropriate dosage forms can be utilized.
Such ingredients and procedures include those described in the following references, each of which is incorporated herein by reference: Powell, M. F. et al., “Compendium of Excipients for Parenteral Formulations” PDA Journal of Pharmaceutical Science & Technology 1998, 52(5), 238-311; Strickley, R. G “Parenteral Formulations of Small Molecule Therapeutics Marketed in the United States (1999)-Part-1” PDA Journal of Pharmaceutical Science & Technology 1999, 53(6), 324-349; and Nema, S. et al., “Excipients and Their Use in Injectable Products” PDA Journal of Pharmaceutical Science & Technology 1997, 51(4), 166-171.
Commonly used pharmaceutical ingredients that can be used as appropriate to formulate the composition for its intended route of administration include:
acidifying agents (examples include but are not limited to acetic acid, citric acid, fumaric acid, hydrochloric acid, nitric acid);
alkalinizing agents (examples include but are not limited to ammonia solution, ammonium carbonate, diethanolamine, monoethanolamine, potassium hydroxide, sodium borate, sodium carbonate, sodium hydroxide, triethanolamine, trolamine);
adsorbents (examples include but are not limited to powdered cellulose and activated charcoa)l;
aerosol propellants (examples include but are not limited to carbon dioxide, CCl2F2, F2ClC—CClF2 and CClF3)
air displacement agents—examples include but are not limited to nitrogen and argon;
antifungal preservatives (examples include but are not limited to benzoic acid, butylparaben, ethylparaben, methylparaben, propylparaben, sodium benzoate);
antimicrobial preservatives (examples include but are not limited to benzalkonium chloride, benzethonium chloride, benzyl alcohol, cetylpyridinium chloride, chlorobutanol, phenol, phenylethyl alcohol, phenylmercuric nitrate and thimerosal);
antioxidants (examples include but are not limited to ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, hypophosphorus acid, monothioglycerol, propyl gallate, sodium ascorbate, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium metabisulfite);
binding materials (examples include but are not limited to block polymers, natural and synthetic rubber, polyacrylates, polyurethanes, silicones, polysiloxanes and styrene-butadiene copolymers);
buffering agents (examples include but are not limited to potassium metaphosphate, dipotassium phosphate, sodium acetate, sodium citrate anhydrous and sodium citrate dihydrate);
carrying agents (examples include but are not limited to acacia syrup, aromatic syrup, aromatic elixir, cherry syrup, cocoa syrup, orange syrup, syrup, corn oil, mineral oil, peanut oil, sesame oil, bacteriostatic sodium chloride injection and bacteriostatic water for injection);
chelating agents (examples include but are not limited to edetate disodium and edetic acid);
colourants (examples include but are not limited to FD&C Red No. 3, FD&C Red No. 20, FD&C Yellow No. 6, FD&C Blue No. 2, D&C Green No. 5, D&C Orange No. 5, D&C Red No. 8, caramel and ferric oxide red);
clarifying agents (examples include but are not limited to bentonite)
emulsifying agents (examples include but are not limited to acacia, cetomacrogol, cetyl alcohol, glyceryl monostearate, lecithin, sorbitan monooleate, polyoxyethylene 50 monostearate);
encapsulating agents (examples include but are not limited to gelatin and cellulose acetate phthalate),
flavourants (examples include but are not limited to anise oil, cinnamon oil, cocoa, menthol, orange oil, peppermint oil and vanillin);
humectants (examples include but are not limited to glycerol, propylene glycol and sorbitol);
levigating agents (examples include but are not limited to mineral oil and glycerin);
oils (examples include but are not limited to arachis oil, mineral oil, olive oil, peanut oil, sesame oil and vegetable oil);
ointment bases (examples include but are not limited to lanolin, hydrophilic ointment, polyethylene glycol ointment, petrolatum, hydrophilic petrolatum, white ointment, yellow ointment, and rose water ointment);
penetration enhancers (transdermal delivery) (examples include but are not limited to monohydroxy or polyhydroxy alcohols, mono- or polyvalent alcohols, saturated or unsaturated fatty alcohols, saturated or unsaturated fatty esters, saturated or unsaturated dicarboxylic acids, essential oils, phosphatidyl derivatives, cephalin, terpenes, amides, ethers, ketones and ureas),
plasticizers (examples include but are not limited to diethyl phthalate and glycerol);
solvents (examples include but are not limited to ethanol, corn oil, cottonseed oil, glycerol, isopropanol, mineral oil, oleic acid, peanut oil, purified water, water for injection, sterile water for injection and sterile water for irrigation);
stiffening agents (examples include but are not limited to cetyl alcohol, cetyl esters wax, microcrystalline wax, paraffin, stearyl alcohol, white wax and yellow wax);
suppository bases (examples include but are not limited to cocoa butter and polyethylene glycols (mixtures));
surfactants (examples include but are not limited to benzalkonium chloride, nonoxynol 10, oxtoxynol 9, polysorbate 80, sodium lauryl sulfate and sorbitan mono-palmitate);
suspending agents (examples include but are not limited to agar, bentonite, carbomers, carboxymethylcellulose sodium, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, kaolin, methylcellulose, tragacanth and veegum);
sweetening agents (examples include but are not limited to aspartame, dextrose, glycerol, mannitol, propylene glycol, saccharin sodium, sorbitol and sucrose);
tablet anti-adherents (examples include but are not limited to magnesium stearate and talc);
tablet binders (examples include but are not limited to acacia, alginic acid, carboxymethylcellulose sodium, compressible sugar, ethylcellulose, gelatin, liquid glucose, methylcellulose, non-crosslinked polyvinyl pyrrolidone, and pregelatinized starch);
tablet and capsule diluents (examples include but are not limited to dibasic calcium phosphate, kaolin, lactose, mannitol, microcrystalline cellulose, powdered cellulose, precipitated calcium carbonate, sodium carbonate, sodium phosphate, sorbitol and starch);
tablet coating agents (examples include but are not limited to liquid glucose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose, ethylcellulose, cellulose acetate phthalate and shellac);
tablet direct compression excipients (examples include but are not limited to dibasic calcium phosphate);
tablet disinteqrants (examples include but are not limited to alginic acid, carboxymethylcellulose calcium, microcrystalline cellulose, polacrillin potassium, cross-linked polyvinylpyrrolidone, sodium alginate, sodium starch glycollate and starch);
tablet glidants (examples include but are not limited to colloidal silica, corn starch and talc);
tablet lubricants (examples include but are not limited to calcium stearate, magnesium stearate, mineral oil, stearic acid and zinc stearate);
tablet/capsule opaquants (examples include but are not limited to titanium dioxide);
tablet polishing agents (examples include but are not limited to carnuba wax and white wax);
thickening agents (examples include but are not limited to beeswax, cetyl alcohol and paraffin);
tonicity agents (examples include but are not limited to dextrose and sodium chloride);
viscosity increasing agents (examples include but are not limited to alginic acid, bentonite, carbomers, carboxymethylcellulose sodium, methylcellulose, polyvinyl pyrrolidone, sodium alginate and tragacanth); and
wetting agents (examples include but are not limited to heptadecaethylene oxycetanol, lecithins, sorbitol monooleate, polyoxyethylene sorbitol monooleate, and polyoxyethylene stearate).
Pharmaceutical compositions according to the present invention can be illustrated as follows:
Sterile i.v. solution: A 5 mg/mL solution of the desired compound of this invention can be made using sterile, injectable water, and the pH is adjusted if necessary. The solution is diluted for administration to 1-2 mg/mL with sterile 5% dextrose and is administered as an i.v. infusion over about 60 minutes.
Lyophilised powder for i.v. administration: A sterile preparation can be prepared with (i) 100-1000 mg of the desired compound of this invention as a lyophilised powder, (ii) 32-327 mg/mL sodium citrate, and (iii) 300-3000 mg Dextran 40. The formulation is reconstituted with sterile, injectable saline or dextrose 5% to a concentration of 10 to 20 mg/mL, which is further diluted with saline or dextrose 5% to 0.2-0.4 mg/mL, and is administered either IV bolus or by IV infusion over 15-60 minutes.
Intramuscular suspension: The following solution or suspension can be prepared, for intramuscular injection:
50 mg/mL of the desired, water-insoluble compound of this invention
5 mg/mL sodium carboxymethylcellulose
4 mg/mL TWEEN 80
9 mg/mL sodium chloride
9 mg/mL benzyl alcohol
Hard Shell Capsules: A large number of unit capsules are prepared by filling standard two-piece hard galantine capsules each with 100 mg of powdered active ingredient, 150 mg of lactose, 50 mg of cellulose and 6 mg of magnesium stearate.
Soft Gelatin Capsules: A mixture of active ingredient in a digestible oil such as soybean oil, cottonseed oil or olive oil is prepared and injected by means of a positive displacement pump into molten gelatin to form soft gelatin capsules containing 100 mg of the active ingredient. The capsules are washed and dried. The active ingredient can be dissolved in a mixture of polyethylene glycol, glycerin and sorbitol to prepare a water miscible medicine mix.
Tablets: A large number of tablets are prepared by conventional procedures so that the dosage unit is 100 mg of active ingredient, 0.2 mg. of colloidal silicon dioxide, 5 mg of magnesium stearate, 275 mg of microcrystalline cellulose, 11 mg. of starch, and 98.8 mg of lactose. Appropriate aqueous and non-aqueous coatings may be applied to increase palatability, improve elegance and stability or delay absorption.
Immediate Release Tablets/Capsules: These are solid oral dosage forms made by conventional and novel processes. These units are taken orally without water for immediate dissolution and delivery of the medication. The active ingredient is mixed in a liquid containing ingredient such as sugar, gelatin, pectin and sweeteners. These liquids are solidified into solid tablets or caplets by freeze drying and solid state extraction techniques. The drug compounds may be compressed with viscoelastic and thermoelastic sugars and polymers or effervescent components to produce porous matrices intended for immediate release, without the need of water.
Dose and Administration
Based upon standard laboratory techniques known to evaluate compounds useful for the treatment of hyper-proliferative disorders and angiogenic disorders, by standard toxicity tests and by standard pharmacological assays for the determination of treatment of the conditions identified above in mammals, and by comparison of these results with the results of known medicaments that are used to treat these conditions, the effective dosage of the compounds of this invention can readily be determined for treatment of each desired indication. The amount of the active ingredient to be administered in the treatment of one of these conditions can vary widely according to such considerations as the particular compound and dosage unit employed, the mode of administration, the period of treatment, the age and sex of the patient treated, and the nature and extent of the condition treated.
The total amount of the active ingredient to be administered will generally range from about 0.001 mg/kg to about 200 mg/kg body weight per day, and preferably from about 0.01 mg/kg to about 20 mg/kg body weight per day. Clinically useful dosing schedules will range from one to three times a day dosing to once every four weeks dosing. In addition, “drug holidays” in which a patient is not dosed with a drug for a certain period of time, may be beneficial to the overall balance between pharmacological effect and tolerability. A unit dosage may contain from about 0.5 mg to about 1500 mg of active ingredient, and can be administered one or more times per day or less than once a day. The average daily dosage for administration by injection, including intravenous, intramuscular, subcutaneous and parenteral injections, and use of infusion techniques will preferably be from 0.01 to 200 mg/kg of total body weight. The average daily rectal dosage regimen will preferably be from 0.01 to 200 mg/kg of total body weight. The average daily vaginal dosage regimen will preferably be from 0.01 to 200 mg/kg of total body weight. The average daily topical dosage regimen will preferably be from 0.1 to 200 mg administered between one to four times daily. The transdermal concentration will preferably be that required to maintain a daily dose of from 0.01 to 200 mg/kg. The average daily inhalation dosage regimen will preferably be from 0.01 to 100 mg/kg of total body weight.
Of course the specific initial and continuing dosage regimen for each patient will vary according to the nature and severity of the condition as determined by the attending diagnostician, the activity of the specific compound employed, the age and general condition of the patient, time of administration, route of administration, rate of excretion of the drug, drug combinations, and the like. The desired mode of treatment and number of doses of a compound of the present invention or a pharmaceutically acceptable salt or ester or composition thereof can be ascertained by those skilled in the art using conventional treatment tests.
Combination Therapies
The compounds of this invention can be administered as the sole pharmaceutical agent or in combination with one or more other pharmaceutical agents where the combination causes no unacceptable adverse effects. Those combined pharmaceutical agents can be other agents having antiproliferative effects such as for example for the treatment of haematological tumours, solid tumours and/or metastases thereof and/or agents for the treatment of undesired side effects. The present invention relates also to such combinations.
Other anti-hyper-proliferative agents suitable for use with the composition of the invention include but are not limited to those compounds acknowledged to be used in the treatment of neoplastic diseases in Goodman and Gilman's The Pharmacological Basis of Therapeutics (Ninth Edition), editor Molinoff et al., publ. by McGraw-Hill, pages 1225-1287, (1996), which is hereby incorporated by reference, especially (chemotherapeutic) anti-cancer agents as defined supra. The combination can be a non-fixed combination or a fixed-dose combination as the case may be.
Methods of testing for a particular pharmacological or pharmaceutical property are well known to persons skilled in the art.
The example testing experiments described herein serve to illustrate the present invention and the invention is not limited to the examples given.
As will be appreciated by persons skilled in the art, the invention is not limited to the particular embodiments described herein, but covers all modifications of said embodiments that are within the spirit and scope of the invention as defined by the appended claims.
The following examples illustrate the invention in greater detail, without restricting it. Further compounds according to the invention, of which the preparation is not explicitly described, can be prepared in an analogous way.
The compounds, which are mentioned in the examples and the salts thereof represent preferred embodiments of the invention as well as a claim covering all subcombinations of the residues of the compound of formula (I) as disclosed by the specific examples.
The term “according to” within the experimental section is used in the sense that the procedure referred to is to be used “analogously to”.
Experimental Part
The following table lists the abbreviations used in this paragraph and in the Intermediate Examples and Examples section as far as they are not explained within the text body.
Other abbreviations have their meanings customary per se to the skilled person. The various aspects of the invention described in this application are illustrated by the following examples which are not meant to limit the invention in any way.
NMR peak forms in the following specific experimental descriptions are stated as they appear in the spectra, possible higher order effects have not been considered. Reactions employing microwave irradiation may be run with a Biotage Initator® microwave oven optionally equipped with a robotic unit. The reported reaction times employing microwave heating are intended to be understood as fixed reaction times after reaching the indicated reaction temperature. The compounds and intermediates produced according to the methods of the invention may require purification. Purification of organic compounds is well known to the person skilled in the art and there may be several ways of purifying the same compound. In some cases, no purification may be necessary. In some cases, the compounds may be purified by crystallization. In some cases, impurities may be stirred out using a suitable solvent. In some cases, the compounds may be purified by chromatography, particularly flash column chromatography, using for example prepacked silica gel cartridges, e.g. from Separtis such as Isolute® Flash silica gel or Isolute® Flash NH2 silica gel in combination with a Isolera® autopurifier (Biotage) and eluents such as gradients of e.g. hexane/ethyl acetate or DCM/methanol. In some cases, the compounds may be purified by preparative HPLC using for example a Waters autopurifier equipped with a diode array detector and/or on-line electrospray ionization mass spectrometer in combination with a suitable prepacked reverse phase column and eluents such as gradients of water and acetonitrile which may contain additives such as trifluoroacetic acid, formic acid or aqueous ammonia. In some cases, purification methods as described above can provide those compounds of the present invention which possess a sufficiently basic or acidic functionality in the form of a salt, such as, in the case of a compound of the present invention which is sufficiently basic, a trifluoroacetate or formate salt for example, or, in the case of a compound of the present invention which is sufficiently acidic, an ammonium salt for example. A salt of this type can either be transformed into its free base or free acid form, respectively, by various methods known to the person skilled in the art, or be used as salts in subsequent biological assays. It is to be understood that the specific form (e.g. salt, free base etc) of a compound of the present invention as isolated as described herein is not necessarily the only form in which said compound can be applied to a biological assay in order to quantify the specific biological activity.
The percentage yields reported in the following examples are based on the starting component that was used in the lowest molar amount. Air and moisture sensitive liquids and solutions were transferred via syringe or cannula, and introduced into reaction vessels through rubber septa. Commercial grade reagents and solvents were used without further purification. The term “concentrated in vacuo” refers to use of a Buchi rotary evaporator at a minimum pressure of approximately 15 mm of Hg. All temperatures are reported uncorrected in degrees Celsius (° C.).
In order that this invention may be better understood, the following examples are set forth. These examples are for the purpose of illustration only, and are not to be construed as limiting the scope of the invention in any manner. All publications mentioned herein are incorporated by reference in their entirety.
Analytical LC-MS Conditions
LC-MS-data given in the subsequent specific experimental descriptions refer (unless otherwise noted) to the following conditions:
Preparative HPLC Conditions
“Purification by preparative HPLC” in the subsequent specific experimental descriptions refers to (unless otherwise noted) the following conditions:
Analytics (Pre- and Post Analytics: Method B):
Preparation:
Chiral HPLC Conditions
If not specified otherwise, chiral HPLC-data given in the subsequent specific experimental descriptions refer to the following conditions:
Analytics:
Preparation:
Flash Column Chromatography Conditions
“Purification by (flash) column chromatography” as stated in the subsequent specific experimental descriptions refers to the use of a Biotage Isolera purification system. For technical specifications see “Biotage product catalogue” on www.biotage.com.
Determination of Optical Rotation Conditions
Optical rotations were measured in dimethyl sulfoxide at 589 nm wavelength, 20° C., concentration 1.0000 g/100 ml, integration time 10 s, film thickness 100.00 mm.
250 mg of 2-[1-(4-ethoxy-2,6-difluorobenzyl)-1H-indazol-3-yl]-5-methoxypyrimidin-4-amine (0.608 mmol, 1 eq.), 608 mg of [1-(tert-butoxycarbonyl)-1H-pyrazol-4-yl]boronic acid and copper (II) acetate were added into a flask and were rinsed with nitrogen. 17 mL chloroform, 0.85 mL triethylamine (6.08 mmol, 10.0 eq.) and 111 mg of N,N-dimethylpyridin-4-amine (0.911 mmol, 1.5 eq.) were added and the reaction mixture was stirred over night at rt. Then the reaction mixture was filtered through celite and washed with dichloromethane. The filtrate was washed with saturated sodiumhydrogencarbonate solution. The aqueous layer was extracted twice with dichloromethane. The combined organic layers were washed with brine, dried over a silicon filter and concentrated in vacuo. The residue was purified by flash chromatography to yield 206 mg of 51% pure target compound, which was used without further purification.
The following intermediates were prepared according to the same procedure from the indicated starting materials (SM=starting material):
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 1.53 (s, 9H), 3.65 (s, 3H), 3.99 (s, 3H), 5.64 (s, 2H), 6.84 (d, 2H), 7.20 (t, 1H), 7.31-7.40 (m, 3H), 7.74 (d,1H), 8.19 (s, 1H), 8.15 (s, 1H), 8.44 (d, 1H), 9.01 (s, 1H), 9.59 (s, 1H).
165 g of 1-(4-ethoxy-2,6-difluorobenzyl)-1H-indazole-3-carboximidamide hydrochloride 1-4-1 (450 mmol, 1.0 eq.), 185 g of 3,3-bis(dimethylamino)-2-methoxypropanenitrile 1-3-1 (1079 mmol, 2.4 eq.) and 19.1 mL of piperidine (225 mmol, 0.5 eq.) were dissolved in 1470 mL of dry 3-methylbutan-1-ol, put under a nitrogen atmosphere and stirred at 110° C. over night. The mixture was cooled down to 0° C. and stirred for crystallization. The resulting suspension was filtered off. The crystals were washed with 1 L hexane and dried in vacuo at 60° C. to provide 65 g (158 mmol, 35%) of the analytically pure target compound.
1H-NMR (400 MHz, DMSO-d6): δ [ppm]=1.26 (t, 3H), 3.84 (s, 3H), 4.00 (q, 2H), 5.60 (s, 2H), 6.66-6.76 (m, 2H), 6.76-6.91 (m, 2H), 7.17 (t, 1H), 7.40 (t, 1H), 7.69 (d, 1H), 7.93 (s, 1H), 8.52 (d, 1H).
The following intermediates were prepared according to the same procedure from the indicated starting materials (SM=starting material):
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 3.85 (s, 3H), 5.73 (s, 2H), 6.85 (br. s., 2H), 7.01-7.13 (m, 2H), 7.15- 7.24 (m, 2H), 7.27- 7.42 (m, 2H), 7.69 (d, 1H), 7.95 (s, 1H), 8.55 (d, 1H).
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 3.66 (s, 3H), 3.85 (s, 3H), 5.59 (s, 2H), 6.74- 6.91 (m, 4H), 7.10-7.24 (m, 3H), 7.35 (dt, 1H), 7.67 (d, 1H), 7.95 (s, 1H), 8.53 (d, 1H).
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 0.80 (t, 3H), 1.48 (sxt, 2H), 3.26-3.33 (m, 2H), 3.85 (s, 3H), 5.62 (s, 2H), 6.83 (br. s., 2H), 7.04-7.21 (m, 5H), 7.35 (ddd, 1H), 7.68 (d, 1H), 7.95 (s, 1H), 8.54 (d, 1H).
360 g of 1-tert-butoxy-N,N,N,N′-tetramethylmethanediamine (Bredereck's reagent) (2068 mmol, 1.0 eq.) and 150 g of methoxyacetonitrile (2068 mmol, 1.0 eq.) were stirred for 18 hours at 80° C. The reaction mixture was concentrated in vacuo. The residue was purified by vacuum distillation (8-23 mmbar; bp 80-83° C.) to yield 117 g (687 mmol, 33%) of the analytical pure target compound as a yellowish liquid.
1H-NMR (400 MHz, DMSO-d6): δ [ppm]=2.23 (s, 6H), 2.29 (s, 6H), 3.23 (d, 1H), 3.36-3.41 (s, 3H), 4.73 (d, 1H).
58 g of ammonium chloride were suspended in 1 L of dry toluene under nitrogen atmosphere and cooled down to 0° C. bath temperature. 541 mL of 2M trimethylaluminium solution in toluene (1083 mmol, 5.0 eq.) were added drop wise. The mixture was stirred at room temperature until disappearance of gassing. 75 g of methyl 1-(4-ethoxy-2,6-difluorobenzyl)-1H-indazole-3-carboxylate 1-5-1 (59.8 mmol, 1.0 eq.) were dissolved in 1 L of dry toluene and added drop wise to the reaction mixture and stirred over night at 80° C. bath temperature. The mixture was cooled down with an ice bath to 0° C. bath temperature, 1.4 L of methanol were added and stirred for one hour at rt. The resulting suspension was filtered over celite and the residue was washed with methanol. The filtrate was concentrated in vacuo and dried in vacuo at 50° C. and the crude product was used without any further purification: 67.3 g (84%).
1H NMR (300 MHz, DMSO-d6) δ [ppm]=1.26 (t, 3H), 4.01 (q, 2H), 5.75 (s, 2H), 6.68-6.78 (m, 2H), 7.34-7.43 (m, 1H), 7.56-7.61 (m, 1H), 7.93 (dd, 2H), 9.29 (br. s, 3H).
The following intermediates were prepared according to the same procedure from the indicated starting materials (SM=starting material):
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 3.67 (s, 3H), 5.73 (s, 2H), 6.76-6.92 (m, 2H), 7.25-7.32 (m, 2H), 7.34- 7.42 (m, 1H), 7.53 (d, 1H), 7.91 (d, 1H), 8.00 (d, 1H), 9.33 (br. s., 4H).
185 g of methyl 1H-indazole-3-carboxylate (1050 mmol, 1.0 eq.) were dissolved in 3 L of dry THF and cooled to 5° C. 411 g of cesium carbonate (1260 mmol, 1.2 eq.) were added stirred for 15 min. 290 g of 2-(bromomethyl)-5-ethoxy-1,3-difluorobenzene (1155 mmol, 1.1 eq.) dissolved in 250 mL THF were added drop wise at 5° C. The precipitate was filtered off. The filtrate was concentrated in vacuo. The residue was crystallized from ethyl acetate/hexane (1:1) to provide 310 g (895 mmol, 85%) of analytically pure target compound.
1H NMR (400 MHz, DMSO-d6) δ [ppm]=1.27 (t, 3H), 3.86 (s, 3H), 4.01 (q, 2H), 5.68 (s, 2H), 6.70-6.76 (m, 2H), 7.32 (t, 1H), 7.50 (t, 1H), 7.84 (d, 1H), 8.00-8.12 (m, 1H).
The following intermediates were prepared according to the same procedure from commercial available starting materials:
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 1.25 (t, 3H), 4.00 (q, 2H), 5.56 (s, 2H), 6.66- 6.76 (m, 2H), 7.14-7.25 (m, 1H), 7.37 (d, 1H), 7.44-7.56 (m, 1H), 7.72 (d, 1H).
1H NMR (300 MHz, DMSO-d6) δ [ppm] = 3.64-3.71 (s, 3 H), 5.70 (s, 2 H) 6.81-6.89 (m, 2 H), 7.22-7.29 (m, 2 H), 7.38 (ddd, 1 H), 7.55 (ddd, 1 H), 7.85 (dt, 1 H) 7.97 (dt, 1 H).
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 5.84 (s, 2H), 7.11-7.45 (m, 5H), 7.58 (ddd, 1H), 7.87 (d, 1H), 7.96 (d, 1H).
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 3.66 (s, 3H), 3.89 (s, 3H), 5.67 (s, 2H), 6.79- 6.90 (m, 2H), 7.20-7.26 (m, 2H), 7.29-7.33 (m, 1H), 7.43-7.47 (m, 1H), 7.84 (d, 1H), 8.05 (dt, 1H).
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 0.73-0.87 (m, 3H), 1.38- 1.58 (m, 2H), 2.42-2.50 (tr, 2H), 5.74 (s, 2H), 7.05-7.15 (m, 2H), 7.15- 7.23 (m, 2H), 7.38 (ddd, 1H), 7.55 (ddd, 1H), 7.85 (d, 1H), 7.91- 7.99 (m, 1H).
1H-NMR (400 MHz, CHLOROFORM-d): δ [ppm] = 1.39 (t, 3H), 3.98 (q, 2H), 5.50 (s, 2H), 6.41-6.50 (m, 2H), 7.39 (d, 1H), 8.47 (d, 1H), 8.94 (d, 1H).
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 3.66 (s, 3H), 5.55 (s, 2H), 6.79-6.87 (m, 2H), 7.16-7.22 (m, 3H), 7.42 (dd, 2H), 7.72 (d, 1H).
1.38 g 1-(4-ethoxy-2,6-difluorobenzyl)-3-iodo-1H-indazole 1-5-2 (3.34 mmol 1.0 eq.), 1.6 g 4-chloro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (6.68 mmol, 2.0 eq.), 240 mg copper(I)bromide (1.67 mmol 0.5 eq.), 1.39 g potassium carbonate (10.0 mmol 3.0 eq.), 122 mg (1,1,bis(diphenylphosphino)ferrocene)-dichloropalladium(II) (0.167 mmol 0.05 eq.) were suspended in 18 mL N,N-dimethylformamide in a sealed tube under argon and stirred overnight at 100° C. The mixture was cooled down, diluted with water and dichlormethane. The aqueous layer was extracted three times with dichloromethane. The combined organic layers were washed with brine, dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash chromatography to yield 0.64 g (1.6 mmol, 48%) of the analytically pure target compound.
1H NMR (300 MHz, DMSO-d6) δ [ppm]=1.25 (t, 3H), 3.99 (q, 2H), 5.66 (s, 2H), 6.67-6.78 (m, 2H), 7.25 (t, 1H), 7.41-7.52 (m, 2H), 7.77 (d, 1H), 7.97 (d, 1H), 8.48 (d, 1H), 8.61-8.68 (m, 1H).
The following intermediates were prepared according to the same procedure from the indicated starting materials (SM=starting material):
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 5.77 (s, 2H), 7.13 (t, 2H), 7.27 (d, 1H), 7.38- 7.53 (m, 3H), 7.81 (d, 1H), 7.96 (d, 1H), 8.46- 8.53 (m, 1H), 8.65 (d, 1H).
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 0.81 (t, 3H), 1.42-1.54 (m, 2H), 2.48-2.56 (m, 2H), 5.69 (s, 2H), 7.06- 7.14 (m, 2H), 7.16-7.28 (m, 3H), 7.36-7.51 (m, 2H), 7.76 (d, 1H), 8.11 (d, 1H), 8.50 (d, 1H), 8.65 (d, 1H).
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 5.77 (s, 2H), 7.02 (d, 1H), 7.21-7.34 (m, 3H), 7.42-7.52 (m, 2H), 7.78 (d, 1H), 8.06 (d, 1H), 8.52 (d, 1H), 8.67 (d, 1H).
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 1.25 (t, 3H), 4.00 (q, 2H), 5.69 (s, 2H), 6.69- 6.78 (m, 2H), 7.55 (dd, 1H), 7.79 (d, 1H), 7.99 (d, 1H), 8.47 (d, 1H), 8.70 (d, 1H), 9.71 (s, 1H).
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 5.79 (s, 2H), 7.07-7.37 (m, 5H), 7.42-7.51 (m, 2H), 7.77 (d, 1H), 8.06 (d, 1H), 8.52 (d, 1H), 8.67 (d, 1H).
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 5.80 (s, 2H), 6.63-6.76 (m, 1H), 6.95-7.09 (m, 1H), 7.22-7.32 (m, 2H), 7.32-7.41 (m, 1H), 7.44 (br. s., 1H), 7.48- 7.58 (m, 1H), 7.82-7.91 (m, 1H), 8.03-8.12 (m, 1H), 8.14-8.25 (m, 1H).
9.25 g of 1-(4-methoxybenzyl)-1H-indazole-3-carbonitrile (1-5-3, 35.1 mmol, 1 eq.) were suspended in 128 ml of dry methanol under a nitrogen atmosphere. 0.949 g (17.6 mmol, 0.5 eq.) of sodium methanolate were added. The reaction mixture was stirred for 18 hours at room temperature. To the resulting mixture were added 2.82 g (52.7 mmol, 1.5 eq.) of ammonium chloride and 1.0 mL (17.6 mmol, 0.5 eq.) of 100% acetic acid and stirred for 5 hours at 50° C. After cooling down to room temperature the mixture was concentrated in vacuo. The residue was partitioned between aq. half saturated sodium hydrogen carbonate solution and dichloromethane/isopropanol 4:1. The aqueous layer was extracted three times with dichloromethane/isopropanol 4:1. The combined organic layers were washed with brine, dried over magnesium sulfate and concentrated in vacuo. The residue was purified by flash chromatography to yield 6.45 g (23 mmol, 65.5%) of the analytically pure target compound.
1H NMR (300 MHz, DMSO-d6) δ [ppm]=3.62-3.70 (s, 3H), 5.57 (s, 2H), 6.37 (br. s., 3H), 6.78-6.88 (m, 2H), 7.10-7.23 (m, 3H), 7.35 (ddd, 1H), 7.68 (d, 1H), 8.27 (d, 1H).
The following intermediates were prepared according to the same procedure from the indicated starting materials (SM=starting material):
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 5.72 (s, 2H), 6.73 (br. s., 3H), 7.01-7.13 (m, 2H), 7.15-7.23 (m, 2H), 7.27- 7.36 (m, 1H), 7.40 (ddd, 1H), 7.69 (d, 1H), 8.27 (d, 1H).
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 0.80 (t, 3H), 1.48 (sxt, 2H), 2.42-2.52 (t, 2H), 5.61 (s, 2H), 6.41 (br. s., 2H), 7.01-7.19 (m, 6H), 7.36 (ddd, 1H), 7.67 (d, 1H), 8.28 (d, 1H).
1.0 g of 1-(4-ethoxy-2,6-difluorobenzyl)-3-iodo-1H-indazole (2.41 mmol, 1.0 eq.) were dissolved in 30 mL dioxane under argon. 344 mg of 6-chloropyrimidin-4-amine (2.66 mmol, 1.1 eq.), 1.81 mL hexabutyldistannane (3.62 mmol, 1.5 eq.) and 847 mg bis(triphenylphosphin)palladium(II)chloride (1.21 mmol, 0.5 eq.) were added. The reaction mixture was stirred over night at 100° C. The reaction mixture was filtered through celite and concentrated in vacuo. The residue was dissolved in ethyl acetate and washed with water and brine, dried over a silicon filter and concentrated in vacuo. The crude product was purified by flash chromatography to yield 104 mg (0.27 mmol, 11%) of the analytically pure target compound.
1H NMR (400 MHz, DMSO-d6) δ [ppm]=1.25 (t, 3H), 4.00 (q, 2H), 5.63 (s, 2H), 6.72 (d, 2H), 6.86 (br. s., 2H), 7.04 (s, 1H), 7.21 (t, 1H), 7.44 (t, 1H), 7.75 (d, 1H), 8.37-8.58 (m, 2H).
188 mg 1-(4-propylbenzyl)-1,2-dihydro-3H-indazol-3-one 1-10-1 (0.706 mmol 1.0 eq.), were suspended in 1.4 mL dichlormethane and 0.163 mL pyridine (1.77 mmol 2.5 eq.). Under nitrogen atmosphere 0.148 mL (0,882 mmol 1.25 eq.) trifluoromethynesulfonic acid were added dropwise at +4° C. After 3 hours at rt, the reaction mixture was filtered off over a silica, concentrated in vacuo to give 275 mg (0.69 mmol, 98%) of Intermediate 1-9-1.
1H NMR (300 MHz, DMSO-d6) δ [ppm]=0.80 (t, 3H), 1.38-1.58 (m, 2H), 2.41-2.44 (m, 2H), 5.60 (s, 2H), 7.04-7.19 (m, 4H), 7.24-7.36 (m, 1H), 7.43-7.59 (m, 1H), 7.71 (d, 1H), 7.84 (d, 1H).
The following intermediates were prepared according to the same procedure from the indicated starting materials (SM=starting material):
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 5.68 (s, 2H), 6.98-7.09 (m, 1H), 7.18-7.37 (m, 3H), 7.56 (ddd, 1H), 7.72 (d, 1H), 7.85 (d, 1H).
231 mg 1,2-dihydro-3H-indazol-3-one (1.72 mmol 1.0 eq.) were dissolved in 2 mL N,N-dimethylformamide at +4° C. 357 mg of potassium carbonate (2.58 mmol 1.5 eq.) was added and then 440 mg 1-(bromomethyl)-4-propylbenzene (2.07 mmol 1.2 eq.) were added portionwise. The reaction was stirred over night at rt. The reaction mixture was diluted with water and ethyl actetate. The layers were separated; the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were dried over a silicon filter and concentrated in vacuo. The residue was purified by crystallisation from methanol to give 188 mg (0.70 mmol, 41%) of Intermediate 1-10-1.
1H NMR (300 MHz, DMSO-d6) δ [ppm]=0.81 (t, 3H), 1.35-1.59 (m, 2H), 2.37-2.44 (m, 2H), 5.27 (s, 2H), 6.94 (t, 1H), 7.06 (s, 4H), 7.21-7.34 (m, 1H), 7.48 (d, 1H), 7.57 (d, 1H), 10.64 (s, 1H).
The following intermediates were prepared according to the same procedure from commercial available starting materials:
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 5.34 (s, 2H), 6.89-7.03 (m, 2H), 7.08-7.24 (m, 2H), 7.32 (ddd, 1H), 7.52 (d, 1H), 7.55-7.62 (m, 1H).
1.30 g of 3-bromo-1-(4-ethoxy-2,6-difluorobenzyl)-1H-pyrazolo[4,3-c]pyridine (3.53 mmol, 1.0 eq.) were dissolved in 6.6 mL dioxane. 1.06 g sodium iodide (7.06 mmol, 2.0 eq.), 161 mg copper iodide (0.85 mmol, 0.24 eq.) and 0.188 mL N,N′-dimethylethylendiamine (1.77 mmol, 0.5 eq.) were added and stirred at rt over night. The reaction mixture was diluted with water and ethyl actetate. The layers were separated, the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were dried over a silicon filter and concentrated in vacuo. The residue was purified by flash chromatography to yield 1.36 g (2.95 mmol, 83%) of the analytically pure target compound.
1H NMR (300 MHz, DMSO-d6) δ [ppm]=1.26 (t, 3H), 4.00 (q, 2H), 5.59 (s, 2H), 6.68-6.76 (m, 2H), 7.72 (d, 1H), 8.46 (d, 1H), 8.71 (d, 1H).
2.69 g 1-(4-methoxybenzyl)-3-(trimethylstannanyl)-1H-indazole 1-13-1 (6.71 mmol 1.0 eq.), 1.30 g 2-bromo-4-chloropyrimidine (6.71 mmol 1.0 eq.), 0.39 g tetrakis(triphenylphosphin)palladium(0) (0.335 mmol 0.05 eq.) were refluxed in 54 mL toluene overnight. The reaction mixture was diluted with water and ethyl actetate. The layers were separated; the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were washed with ammoniumchlorid-solution dried over a silicon filter and concentrated in vacuo. The residue was purified by flash chromatography to yield 814 mg (2.3 mmol, 34%) of the analytically pure target compound.
1H NMR (300 MHz, DMSO-d6) δ [ppm]=3.66 (s, 3H), 5.71 (s, 2H), 6.85 (d, 2H), 7.20-7.35 (m, 3H), 7.45 (t, 1H), 7.61 (d, 1H), 7.82 (d, 1H), 8.46 (d, 1H), 8.88 (d, 1H).
5.0 g 3-iodo-1-(4-methoxybenzyl)-1H-indazole (13.7 mmol 1.0 eq.), 6.30 g Hexamethyldistannane (19.2 mMol, 1.4 eq.), 0.79 g tetrakis(triphenylphosphin)-palladium(0) (0.69 mmol, 0.05 eq.) were dissolved in 500 mL dioxane under argon and stirred at +100<C over night. The reaction mixture was diluted with 50 mL potassium fluoride solution halfconcentrated and ethyl actetate. The layers were separated; the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were dried over a silicon-filter and concentrated in vacuo. The residue was purified by flash chromatography to yield 2.69 g (6.24 mmol, 45%) of the analytically pure target compound.
1H NMR (400 MHz, DMSO-d6) δ [ppm]=0.36 (s, 9H), 3.65 (s, 3H), 5.57 (s, 2H), 6.76-6.86 (m, 2H), 7.00-7.09 (m, 1H), 7.11-7.19 (m, 2H), 7.28 (t, 1H), 7.66 (d, 1H), 7.59 (d, 1H).
100 mg 2-[1-(4-ethoxy-2,6-difluorobenzyl)-1H-indazol-3-yl]-5-methoxypyrimidin-4-amine (0.243 mmol 1.0 eq.), 59.3 mg phenylboronic acid (0.486 mmol, 2.0 eq.), 180 mg copper(II) acetate (0.972 mmol, 4.0 eq.), were stirred in chloroform at rt. 0.136 mL triethylamine (0.972 mmol, 4.0 eq.), 14.8 mg 4-dimethylaminopyridine (0.122 mmol, 0.5 eq.) were added, stirred 22 h at rt and 22 h at +60° C. After filtration, the reaction mixture was concentrated in vacuo. The residue was purified by flash chromatography to yield 2 mg (1.7%) of example 2-1-1.
LC-MS: Rt=1.28 min; MS (ESIpos) m/z=487 [M+H]+.
The following compounds were prepared according to the same procedure from the indicated starting materials (SM=starting material):
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 3.66 (s, 3H), 4.01 (s, 3H), 5.62 (s, 2H), 6.79- 6.89 (m, 2H), 7.07-7.18 (m, 1H), 7.22-7.30 (m, 2H), 7.31-7.41 (m, 2H), 7.73 (d, 1H), 8.21-8.28 (m, 2H), 8.32 (d, 1H), 8.42-8.50 (m, 1H), 9.13 (d, 1H), 9.21 (s, 1H).
1H-NMR (400 MHz, METHANOL- d4): δ [ppm] = [ppm] = 3.61 (br. s., 3H), 3.72 (br. s., 3H), 4.01 (br. s., 3H), 5.46 (br. s., 2H), 6.32 (br. s., 1H), 6.63 (br. s., 2H), 7.02 (br. s., 2H), 7.13 (br. s., 1H), 7.41 (br. s., 1H), 7.59 (br. s., 2H), 7.83 (br. s., 1H).
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 3.67 (s, 3H), 4.00 (s, 3H), 5.61 (s, 2H), 6.80- 6.89 (m, 2H), 7.01-7.17 (m, 2H), 7.22-7.42 (m, 5H), 7.72 (d, 1H), 7.92- 8.02 (m, 2H), 8.18 (s, 1H), 8.36 (d, 1H), 8.92 (s, 1H).
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 3.66 (s, 3H), 3.98 (s, 3H), 5.61 (s, 2H), 6.78- 6.90 (m, 2H), 7.07-7.22 (m, 3H), 7.22-7.30 (m, 2H), 7.35 (t, 1H), 7.72 (d, 1H), 7.96 (dd, 2H), 8.17 (s, 1H), 8.31 (d, 1H), 9.04 (s, 1H).
150 mg 2-[1-(4-ethoxy-2,6-difluorobenzyl)-1H-indazol-3-yl]-5-methoxypyrimidin-4-amine (0.365 mmol 1.0 eq.), 131 mg 4-bromopyridazine hydrobromide (0.547 mmol 1.5 eq.), 356 mg cesium carbonate (1.09 mmol, 3.0 eq.), 31.6 mg 4,5-bis-(diphenylphosphino)-9,9-dimethylxanthene (0.55 mmol, 0.15 eq.), 8.2 mg palladium(II) acetate (0.036 mmol, 0.1 eq.) were stirred in 4.8 mL dioxane under nitrogen in a sealed tube at +100° C. over night. The reaction mixture was cooled down to room temperature, filtratered off and concentrated in vacuo. The residue was purified by flash chromatography to yield 72.7 mg (0.15 mmol, 41%) of the analytically pure target compound.
1H NMR (500 MHz, DMSO-d6) δ [ppm]=1.30 (t, 3H), 4.00-4.12 (m, 5H), 5.70 (s, 2H), 6.83 (d, 2H), 7.29 (t, 1H), 7.46-7.57 (m, 1H), 7.87 (d, 1H), 8.39-8.51 (m, 2H), 8.79 (dd, 1H), 8.90-8.97 (m, 1H), 9.68 (d, 1H), 9.80 (s, 1H).
The following compounds were prepared according to the same procedure from the indicated starting materials (SM=starting material):
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 1.25 (t, 3H), 3.97-4.06 (m, 5H), 5.65 (s, 2H), 6.79 (d, 2H), 7.26 (t, 1H), 7.47 (t, 1H), 7.82 (d, 1H), 8.38-8.49 (m, 2H), 8.59 (d, 1H), 8.70 (dd, 1H), 8.84 (s, 1H), 9.08 (s, 1H).
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 1.26 (t, 3H), 4.02 (q, 2H), 5.70 (s, 2H), 6.70- 6.81 (m, 2H), 7.24-7.33 (m, 1H), 7.45-7.53 (m, 1H), 7.81 (d, 1H), 7.88 (dd, 1H), 8.35 (d, 1H), 8.50-8.58 (m, 2H), 8.79 (d, 1H), 8.92 (d, 1H), 10.67 (s, 1H).
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 1.28 (t, 3H), 3.95 (s, 3H), 4.03 (q, 2H), 5.66 (s, 2H), 6.72-6.82 (m, 2H), 6.84-6.95 (m, 1H), 7.19-7.32 (m, 2H), 7.46 (td, 1H), 7.81 (d, 1H), 7.99 (dd, 1H), 8.10 (s, 1H), 8.51 (d, 1H), 9.20 (d, 1H), 14.50 (br. s, 1H).
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 1.29 (t, 3H), 3.95 (s, 3H), 4.04 (q, 2H), 5.65 (s, 2H), 6.72-6.84 (m, 2H), 6.93-7.04 (m, 1H), 7.25 (t, 1H), 7.41-7.51 (m, 1H), 7.69 (dd, 1H), 7.81 (d, 1H), 8.10 (s, 1H), 8.48 (d, 1H), 9.26 (dd, 1H), 14.35 (s, 1H).
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 1.28 (t, 3H), 2.83 (d, 3H), 3.96-4.12 (m, 5H), 5.68 (s, 2H), 6.66-6.82 (m, 2H), 7.24 (t, 1H), 7.37-7.56 (m, 1H), 7.80 (d, 1H), 8.36-8.50 (m, 2H), 8.75 (d, 1H), 8.85 (s, 1H), 8.92 (br. s., 1H), 10.04 (s, 1H).
100 mg 5-methoxy-2-[1-(4-propylbenzyl)-1H-indazol-3-yl]pyrimidin-4-amine (0.27 mmol, 1.0 eq.), 101 mg 4-chlorpyprimidine hydrochloride (0.67 mmol, 2.5 eq.), 77 mg sodium-tert-butylat (0.80 mmol, 3.0 eq.), 83.4 mg (R)-(+)-2,2′-bis-(diphenylphosphino)-1,1′-binaphthyl (0.134 mmol, 0.5 eq.), 24.5 mg tris-(dibenzylidenaceton)dipalladium (0.027 mmol, 0.1 eq.) were suspended in 2 mL N,N-dimethylformamide and stirred at 100° C. over night under nitrogen. The reaction mixture was cooled to room temperature, water and dichloromethane were added and the aqueous phase was extracted with dichloromethane twice. The combined organic phases were dried over a silica filter and concentrated in vacuo. Flash chromatography yielded 146 mg of the impure product. Preparative thin layer chromatography yielded 32 mg (0.071 mmol, 26%) Example 2-3-1 and a mother liquor, which after purification by HPLC gave additional 23 mg (0.051 mmol, 19%) Example 2-3-1.
1H-NMR (400 MHz, DMSO-d6): δ [ppm]=0.80 (t, 3H), 1.49 (sxt, 2H), 2.43-2.51 (m, 2H), 4.00 (s, 3H), 5.70 (s, 2H), 7.12 (d, 2H), 7.20-7.29 (m, 3H), 7.41 (t, 1H), 7.77 (d, 1H), 8.40-8.53 (m, 2H), 8.55-8.68 (m, 2H), 8.84 (s, 1H), 9.18 (s, 1H).
The following compounds were prepared according to the same procedure from the indicated starting materials (SM=starting material):
127 mg of N-(4-{[tert-butyl(dimethyl)silyl]oxy}phenyl)-2-[1-(4-ethoxy-2,6-difluorobenzyl)-1H-indazol-3-yl]-5-methoxypyrimidin-4-amine (0.205 mmol, 1.0 eq.) were suspended in 4 mL dioxane. 3.3 mL Hydrochlorid acid (4 M in dioxane) were added and stirred over night at rt. The solution was partitioned between aqueous half saturated sodium hydrogen carbonate solution and dichloro-methane/isopropanol 4:1. The aqueous layer was extracted three times with dichloromethane/isopropanol 4:1. The combined organic layers were washed with brine, dried over magnesium sulfate and concentrated in vacuo. The residue was purified by flash chromatography NH2-column to yield 48.8 mg (0.09 mmol, 45%) of the analytically pure target compound.
1H NMR (400 MHz, DMSO-d6) δ [ppm]=1.28 (t, 3H), 3.97 (s, 3H), 4.03 (q, 2H), 5.63 (s, 2H), 6.70-6.83 (m, 4H), 7.14 (t, 1H), 7.43 (ddd, 1H), 7.69-7.78 (m, 3H), 8.09 (s, 1H), 8.35 (d, 1H), 8.69 (s, 1H), 9.17 (s, 1H).
100 mg 3-(4-chloropyridin-2-yl)-1-(4-ethoxy-2,6-difluorobenzyl)-1H-indazole (0.25 mmol, 1.0 eq.), 35.7 mg 4-bromopyridazine hydrobromide (0.375 mmol, 1.5 eq.), 245 mg cesium carbonate (0.75 mmol, 3.0 eq.), 22 mg 4,5-bis(diphenyl-phosphino)-9,9-dimethylxanthene (0.038 mmol, 0.15 eq.), 5.6 mg palladium(II) acetate (0.025 mmol, 0.1 eq.) were stirred in 3.2 mL dioxane under nitrogen in a sealed tube at +100° C. over night. The reaction mixture was cooled, filtered over silica and the filtrate was concentrated in vacuo. The residue was purified by HPLC to yield 7 mg (0.02 mmol, 6.1%) of the analytically pure target compound.
1H NMR (400 MHz, DMSO-d6) δ [ppm]=1.26 (t, 3H), 4.01 (q, 2H), 5.64 (s, 2H), 6.69-6.80 (m, 2H), 6.90 (d, 1H), 7.22 (t, 1H), 7.44 (t, 1H), 7.75 (d, 1H), 7.92 (dd, 1H), 8.23 (d, 1H), 8.39 (d, 1H), 8.47-8.57 (m, 2H), 8.73 (s, 1H), 10.13 (s, 1H).
The following compounds were prepared according to the same procedure from the indicated starting materials (SM=starting material):
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 1.26 (t, 3H), 4.01 (q, 2H), 5.64 (s, 2H), 6.56- 6.83 (m, 2H), 7.22 (t, 1H), 7.44 (t, 1H), 7.74 (d, 1H), 7.81 (dd, 1H), 8.42 (d, 1H), 8.48-8.58 (m, 2H), 8.86 (s, 2H), 10.73 (s, 1H).
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 1.25 (t, 3H), 4.00 (q, 2H), 5.62 (s, 2H), 6.69- 6.75 (m, 2H), 6.90 (dd, 1H), 7.19 (t, 1H), 7.35- 7.45 (m, 1H), 7.55 (d, 1H), 7.72 (d, 1H), 8.34 (d, 1H), 8.44-8.54 (m, 2H), 8.60 (s, 1H), 9.32 (s, 1H).
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 1.26 (s, 3H), 4.01 (q, 2H), 5.68 (s, 2H), 6.69- 6.80 (m, 2H), 6.91 (dd, 1H), 7.77 (d, 1H), 7.99 (dd, 1H), 8.24-8.29 (m, 1H), 8.41 (d, 1H), 8.44 (d, 1H), 8.56 (d, 1H), 8.70-8.79 (m, 1H), 9.75 (s, 1H), 10.19 (s, 1H).
1H-NMR (500 MHz, DMSO-d6): δ [ppm] = 3.71 (s, 3H), 5.66-5.74 (m, 4H), 6.86-6.97 (m, 2H), 7.27-7.36 (m, 4H), 7.48 (ddd, 1H), 7.87 (d, 1H), 8.08 (br. s., 2H), 8.51 (d, 1H), 8.85 (d, 1H).
120 mg 3-(4-chloropyridin-2-yl)-1-(4-ethoxy-2,6-difluorobenzyl)-1H-indazole (0.3 mmol 1.0 eq.), 84.7 mg 1-(ethoxymethyl)-1H-pyrazol-4-amine (0.6 mmol, 2.0 eq.), 86.5 mg sodium-tert-butylat (0.9 mmol 3.0 eq.), 93.4 mg (R)-(+)-2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (0.150 mmol, 0.5 eq.), 27.5 mg tris(dibenzylidenaceton)dipalladium (0.03 mmol, 0.1 eq.), were suspended in 1.6 mL N,N-dimethylformamid and stirred 6 h at 100° C. under nitrogen. Once more all reactants were added and stirred at +100° C. over night. The reaction mixture was cooled to room temperature, put into water and the aqueous layer was extracted three times with dichloromethane/isopropanol 4:1. The combined organic layers were washed with brine, dried over magnesium sulfate and concentrated in vacuo. The residue was purified by flash chromatography to yield 56 mg (0.11 mmol, 37%) of the analytically pure target compound.
1H NMR (400 MHz, DMSO-d6) δ [ppm]=1.07 (t, 3H), 1.21-1.28 (m, 3H), 3.49 (q, 2H), 3.95-4.05 (m, 3H), 5.37 (s, 2H), 5.58-5.63 (m, 2H), 6.65 (dd, 1H), 6.68-6.76 (m, 2H), 7.11-7.21 (m, 1H), 7.36-7.45 (m, 2H), 7.45-7.51 (m, 1H), 7.69 (d, 1H), 7.90-7.93 (m, 1H), 8.19-8.26 (m, 1H), 8.44 (s, 1H), 8.50 (d, 1H).
The following compounds were prepared according to the same procedure from the indicated starting materials (SM=starting material):
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 5.75 (s, 2H), 6.89 (dd, 1H), 7.13 (t, 2H), 7.24 (d, 1H), 7.41-7.51 (m, 2H), 7.79 (d, 1H), 7.94 (dd, 1H), 8.20 (d, 1H), 8.39 (d, 1H), 8.48-8.59 (m, 2H), 8.73 (s, 1H), 10.15 (s, 1H).
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 0.81 (t, 3H), 1.49 (sxt, 2H), 2.42-2.52 (m, 2H), 5.68 (s, 2H), 6.89 (dd, 1H), 7.06-7.13 (m, 2H), 7.14-7.25 (m, 3H), 7.34- 7.46 (m, 1H),) 7.73 (d, 1H), 7.93 (dd, 1H), 8.32 (d, 1H), 8.39 (d, 1H), 8.47-8.61 (m, 2H), 8.76 (s, 1H), 10.13 (s, 1H).
56 mg 2-[1-(4-ethoxy-2,6-difluorobenzyl)-1H-indazol-3-yl]-N-[1-(ethoxymethyl)-1H-pyrazol-4-yl]pyridin-4-amine (0.111 mmol 1.0 eq.), were stirred with 1 mL acetic acid and 0.5 mL conc hydrochloric acid for 45 min at +90° C. in a sealed tube. The reaction mixture was add to water, the crystals were filtered off and were purified by HPLC to yield 5 mg (0.01 mmol, 10%) of the analytically pure target compound.
1H NMR (400 MHz, DMSO-d6) δ [ppm]=1.28 (t, 3H), 4.03 (q, 2H), 5.63 (s, 2H), 6.63 (dd, 1H), 6.69-6.79 (m, 2H), 7.19 (t, 1H), 7.38-7.52 (m, 3H), 7.66-7.80 (m, 2H), 8.22 (d, 1H), 8.33 (s, 1H), 8.52 (d, 1H), 12.77 (br. s., 1H).
50 mg of 3-(4-chloropyridin-2-yl)-1-(4-ethoxy-2,6-difluorobenzyl)-1H-indazole (0.125 mmol, 1.0 eq.) and 140 mg of 1-ethyl-1H-1,2,4-triazol-5-amine (1.25 mmol, 10 eq.) were dissolved in 0.6 mL NMP and were heated in a microwave oven up to 200° C. over 6 h. The reaction mixture was diluted with water and dichloromethane, filtered through a silicone filter and concentrated in vacuo. Preparative HPLC purification provided 4 mg (0.01 mmol, 6%) of the analytically pure target compound.
1H-NMR (400 MHz, DMSO-d6): δ [ppm]=1.22-1.33 (m, 6H), 4.00 (q, 2H), 4.14 (q, 2H), 5.64 (s, 2H), 6.70-6.76 (m, 2H), 7.17-7.26 (m, 1H), 7.40-7.47 (m, 1H), 7.67-7.76 (m, 3H), 8.08 (d, 1H), 8.44 (d, 1H), 8.52-8.58 (m, 1H), 9.55 (s, 1H).
The following compounds were prepared according to the same procedure from the indicated starting materials (SM=starting material):
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 1.25 (t, 3H), 3.99 (q, 2H), 5.64 (s, 2H), 6.66- 6.80 (m, 2H), 7.14-7.23 (m, 1H), 7.36-7.46 (m, 1H), 7.47-7.55 (m, 1H), 7.65-7.73 (m, 1H), 8.11- 8.18 (m, 1H), 8.33- 8.40 (m, 1H), 8.52 (d, 1H), 9.63 (br. s., 1H).
273 mg of 2-[1-(2-fluorobenzyl)-1H-indazol-3-yl]-5-methoxy-N-(pyrimidin-4-yl)-pyrimidin-4-amine (2-3-2, 0.64 mmol, 1.0 eq.) were dissolved in 10 mL NMP. 249 mg of sodium sulfide (3.19 mmol, 5.0 eq.) were added and the reaction mixture was stirred at 140° C. for 3.5 h. Half saturated ammonium chloride solution and ethyl acetate were added. A resulting precipitate was dissolved in methanol, dried with magnesium sulfate and concentrated in vacuo. The residue was purified by flash chromatography and preparative HPLC to yield 16 mg (0.03 mmol, 6%) of the analytically pure target compound.
1H-NMR (300 MHz, DMSO-d6): δ [ppm]=5.77 (s, 2H), 7.12-7.27 (m, 4H), 7.29-7.38 (m, 1H), 7.39-7.49 (m, 1H), 7.71-7.84 (m, 1H), 8.24 (s, 1H), 8.39-8.45 (m, 1H), 8.45-8.52 (m, 1H), 8.56 (d, 1H), 8.82 (s, 1H), 9.19 (br. s., 1H), 11.25 (br. s., 1H).
To 9.9 mg tert-butyl 4-({5-methoxy-2-[1-(4-methoxybenzyl)-1H-indazol-3-yl]-pyrimidin-4-yl}amino)-1H-pyrazole-1-carboxylate (0.019 mmol, 1 eq.) stirred in 1 mL dichlormethane at +4° C. were added 0.029 mL trifluoroacetic acid (0.375 mmol, 20 eq.). The reaction mixture was stirred at room temperature over night. The reaction mixture was added to sodium carbonate solution and dichlormethane, stirred for 30 min and the organic phase was separated. The aqueous phase was washed three times with dichlormethane. The combined organic layers were washed with brine, dried over magnesium sulfate and concentrated in vacuo to provide 13.4 mg (0.03 mmol, 8%) of the analytically pure target compound.
1H NMR (300 MHz, DMSO-d6) δ [ppm]=3.65 (s, 3H), 3.96 (s, 3H), 5.63 (s, 2H), 6.78-6.94 (m, 2H), 7.12-7.23 (m, 1H), 7.25-7.32 (m, 2H), 7.33-7.42 (m, 1H), 7.75 (d, 1H), 7.86 (br. s., 1H), 8.07 (s, 1H), 8.42 (d, 2H), 9.26 (s, 1H), 12.57 (br. s., 1H).
The following compounds were prepared according to the same procedure from the indicated starting materials (SM=starting material):
1H-NMR (300 MHz, DMSO-d6): δ [ppm] = 1.25 (t, 3H), 3.99 (q, 2H), 5.64 (s, 2H), 6.66- 6.80 (m, 2H), 7.14-7.23 (m, 1H), 7.36-7.46 (m, 1H), 7.47-7.55 (m, 1H), 7.65-7.73 (m, 1H), 8.11- 8.18 (m, 1H), 8.33- 8.40 (m, 1H), 8.52 (d, 1H), 9.63 (br. s., 1H).
38.9 mg 2-[1-(2,4-difluorobenzyl)-1H-indazol-3-yl]pyridin-4-amine (0.116 mmol, 1.0 eq.), 41.6 mg 4-bromopyrimidine hydrobromide (0.173 mmol, 1.5 eq.), 26.2 mg 4-chloroyrimidine hydrochloride (0.173 mmol, 1.5 eq.), 79.9 mg potassium carbonate (0.578 mmol, 5.0 eq.) and 3 mL N,N-dimethylformamide were stirred over night at +100° C. in a sealed tube. The reaction mixture was diluted with 3 mL dichlormethane, filtrated through an aluminiumoxide column and concentrated in vacuo. The residue was purified by HPLC to yield 4.9 mg (0.01 mmol, 10%) of the analytically pure target compound.
1H NMR (400 MHz, DMSO-d6) δ [ppm]=5.75 (s, 2H), 6.89 (dd, 1H), 6.97-7.09 (m, 1H), 7.19-7.32 (m, 3H), 7.36-7.51 (m, 1H), 7.76 (d, 1H), 7.92 (dd, 1H), 8.28 (d, 1H), 8.39 (d, 1H), 8.49-8.61 (m, 2H), 8.75 (s, 1H), 10.13 (s, 1H).
135 mg of 2-({2-[1-(4-ethoxy-2,6-difluorobenzyl)-1H-indazol-3-yl]-5-methoxypyrimidin-4-yl}amino)benzoic acid 2-2-4 (0.254 mmol, 1.0 eq.) were dissolved in 2.9 mL N,N-dimethylformamide. 0.27 ml of ammonia solution (7M in methanol, 1.9 mmol, 7.5 eq.), 0.17 mL N,N-diisopropylethylamine (1.0 mmol, 4.0 eq.) and 145 mg of Benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphat (0.28 mmol, 1.1 eq.) were added and stirred over night at rt. To the reaction mixture water was added. A yellowish precipitated was formed, washed with water and dried at 50° C. over 3 days under vacuum. The crude product was purified by chromatography to provide 58 mg of the 91% pure target compound: 0.10 mmol, 39%.
1H NMR (400 MHz, DMSO-d6) δ [ppm]=1.30 (t, 3H), 3.97-4.07 (m, 5H), 5.58 (s, 2H), 6.68-6.76 (m, 2H), 7.19-7.26 (m, 1H), 7.46-7.56 (m, 2H), 7.76-7.85 (m, 4H), 7.89-7.96 (m, 2H).
The following compounds were prepared according to the same procedure from the indicated starting materials (SM=starting material):
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 1.29 (t, 3H), 3.09 (d, 3H), 3.44 (s, 3H), 4.04 (q, 2H), 5.72 (s, 2H), 6.71-6.80 (m, 2H), 7.27- 7.37 (m, 2H), 7.41- 7.47 (m, 1H), 7.53-7.60 (m, 1H), 7.64 (d, 1H), 7.70-7.80 (m, 2H), 7.90 (d, 1H), 8.09 (dd, 1H), 8.35-8.44 (m, 1H), 14.41 (s, 1H).
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 1.29 (t, 3H), 3.09 (d, 3H), 3.43 (s, 3H), 4.03 (q, 2H), 5.72 (s, 2H), 6.70-6.81 (m, 2H), 7.29 (s, 1H), 7.56 (td, 1H), 7.62-7.69 (m, 1H), 7.69- 7.79 (m, 3H), 7.90 (d, 1H), 8.36-8.46 (m, 1H), 14.57 (s, 1H).
1H-NMR (400 MHz, DMSO-d6): δ [ppm] = 1.28 (t, 3H), 3.91-4.08 (m, 5H), 5.66 (s, 2H), 6.70-6.86 (m, 2H), 7.20- 7.36 (m, 2H), 7.42- 7.54 (m, 1H), 7.70 (dd, 1H), 7.75-7.86 (m, 2H), 8.25 (s, 1H), 8.31 (br. s., 1H), 8.44 (d, 1H), 9.30 (dd, 1H), 11.71 (s, 1H).
Biological Investigations
The following assays can be used to illustrate the commercial utility of the compounds according to the present invention.
Examples were tested in selected biological assays one or more times. When tested more than once, data are reported as either average values or as median values, wherein
Examples were synthesized one or more times. When synthesized more than once, data from biological assays represent average values calculated utilizing data sets obtained from testing of one or more synthetic batch.
Biological Assay 1.0:
Bub1 Kinase Assay
Bub1-inhibitory activities of compounds described in the present invention were quantified using a time-resolved fluorescence energy transfer (TR-FRET) kinase assay which measures phosphorylation of the synthetic peptide Biotin-Ahx-VLLPKKSFAEPG (C-terminus in amide form), purchased from e.g. Biosyntan (Berlin, Germany) by the (recombinant) catalytic domain of human Bub1 (amino acids 704-1085), expressed in Hi5 insect cells with an N-terminal His6-tag and purified by affinity-(Ni-NTA) and size exclusion chromatography.
In a typical assay 11 different concentrations of each compound (0.1 nM, 0.33 nM, 1.1 nM, 3.8 nM, 13 nM, 44 nM, 0.15 μM, 0.51 μM, 1.7 μM, 5.9 μM and 20 μM) were tested in duplicate within the same microtiter plate. To this end, 100-fold concentrated compound solutions (in DMSO) were previously prepared by serial dilution (1:3.4) of 2 mM stocks in a clear low volume 384-well source microtiter plate (Greiner Bio-One, Frickenhausen, Germany), from which 50 nl of compounds were transferred into a black low volume test microtiter plate from the same supplier. Subsequently, 2 μL of Bub1 (the final concentration of Bub1 was adjusted depending on the activity of the enzyme lot in order to be within the linear dynamic range of the assay: typically 200 ng/mL were used) in aqueous assay buffer [50 mM Tris/HCl pH 7.5, 10 mM magnesium chloride (MgCl2), 200 mM potassium chloride (KCl), 1.0 mM dithiothreitol (DTT), 0.1 mM sodium ortho-vanadate, 1% (v/v) glycerol, 0.01% (w/v) bovine serum albumine (BSA), 0.005% (v/v) Trition X-100 (Sigma), 1× Complete EDTA-free protease inhibitor mixture (Roche)] were added to the compounds in the test plate and the mixture was incubated for 15 min at 22° C. to allow pre-equilibration of the putative enzyme-inhibitor complexes before the start of the kinase reaction, which was initiated by the addition of 3 μL 1.67-fold concentrated solution (in assay buffer) of adenosine-tri-phosphate (ATP, 10 μM final concentration) and peptide substrate (1 μM final concentration). The resulting mixture (5 μL final volume) was incubated at 22° C. during 60 min., and the reaction was stopped by the addition of 5 μL of an aqueous EDTA-solution (50 mM EDTA, in 100 mM HEPES pH 7.5 and 0.2% (w/v) bovine serum albumin) which also contained the TR-FRET detection reagents (0.2 μM streptavidin-XL665 [Cisbio Bioassays, Codolet, France] and 1 nM anti-phosho-Serine antibody [Merck Millipore, cat. #35-001] and 0.4 nM LANCE EU-W1024 labeled anti-mouse IgG antibody [Perkin-Elmer, product no. AD0077, alternatively a Terbium-cryptate-labeled anti-mouse IgG antibody from Cisbio Bioassays can be used]). The stopped reaction mixture was further incubated 1 h at 22° C. in order to allow the formation of complexes between peptides and detection reagents. Subsequently, the amount of product was evaluated by measurement of the resonance energy transfer from the Eu-chelate-antibody complex recognizing the Phosphoserine residue to the streptavidin-XL665 bound to the biotin moiety of the peptide. To this end, the fluorescence emissions at 620 nm and 665 nm after excitation at 330-350 nm were measured in a TR-FRET plate reader, e.g. a Rubystar or Pherastar (both from BMG Labtechnologies, Offenburg, Germany) or a Viewlux (Perkin-Elmer) and the ratio of the emissions (665 nm/622 nm) was taken as indicator for the amount of phosphorylated substrate. The data were normalised using two sets of (typically 32-) control wells for high—(=enzyme reaction without inhibitor=0%=Minimum inhibition) and low—(=all assay components without enzyme=100%=Maximum inhibition) Bub1 activity. IC50 values were calculated by fitting the normalized inhibition data to a 4-parameter logistic equation (Minimum, Maximum, IC50, Hill; Y=Max+(Min−Max)/(1+(X/IC50)Hill)).
Biological Assay 2.0:
Proliferation Assay:
Cultivated tumor cells (cells were ordered from ATCC, except HeLa-MaTu and HeLa-MaTu-ADR, which were ordered from EPO-GmbH, Berlin) were plated at a density of 1000 to 5000 cells/well, depending on the growth rate of the respective cell line, in a 96-well multititer plate in 200 μL of their respective growth medium supplemented 10% fetal calf serum. After 24 hours, the cells of one plate (zero-point plate) were stained with crystal violet (see below), while the medium of the other plates was replaced by fresh culture medium (200 μL), to which the test substances were added in various concentrations (0 μM, as well as in the range of 0.001-10 μM; the final concentration of the solvent dimethyl sulfoxide was 0.5%). The cells were incubated for 4 days in the presence of test substances. Cell proliferation was determined by staining the cells with crystal violet: the cells were fixed by adding 20 μl/measuring point of an 11% glutaric aldehyde solution for 15 minutes at room temperature. After three washing cycles of the fixed cells with water, the plates were dried at room temperature. The cells were stained by adding 100 μl/measuring point of a 0.1% crystal violet solution (pH 3.0). After three washing cycles of the stained cells with water, the plates were dried at room temperature. The dye was dissolved by adding 100 μl/measuring point of a 10% acetic acid solution. Absorption was determined by photometry at a wavelength of 595 nm. The change of cell number, in percent, was calculated by normalization of the measured values to the absorption values of the zero-point plate (=0%) and the absorption of the untreated (0 μm) cells (=100%). The IC50 values were determined by means of a 4 parameter fit using the company's own software.
The following table gives the data regarding Bub1 kinase inhibition, and inhibition of HeLa cell proliferation, for the examples of the present invention for the biological assays 1 and 2:
| Number | Date | Country | Kind |
|---|---|---|---|
| 13160520.6 | Mar 2013 | EP | regional |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2014/055657 | 3/20/2014 | WO | 00 |
