Source: https://patents.justia.com/patent/9988378
Timestamp: 2019-06-17 01:22:12
Document Index: 364775373

Matched Legal Cases: ['Application No. 61', 'Application No. 201380042831', 'Application No. 102138784', 'Application No. 01380042831', 'Application No. 2015', 'Application No. 2015117950']

US Patent for 1 H-pyrazole and 4,5-disubstituted thiazole inhibitors of SYK Patent (Patent # 9,988,378 issued June 5, 2018) - Justia Patents Search
Justia Patents Heterocyclic Carbon Compounds Containing A Hetero Ring Having Chalcogen (i.e., O,s,se Or Te) Or Nitrogen As The Only Ring Hetero Atoms DoaiUS Patent for 1 H-pyrazole and 4,5-disubstituted thiazole inhibitors of SYK Patent (Patent # 9,988,378)
1 H-pyrazole and 4,5-disubstituted thiazole inhibitors of SYK
Oct 23, 2013 - Hoffmann-La Roche Inc.
The present invention relates to the use of novel compounds of formula (I), wherein all variable substituents are defined as described herein, which are SYK inhibitors and are useful for the treatment of auto-immune and inflammatory diseases.
This application is a National Stage Application of PCT/EP2013/072132 filed Oct. 23, 2013, which claims priority from U.S. Provisional Patent Application No. 61/718,740, filed on Oct. 26, 2012. The priority of both said PCT and U.S. Provisional Patent Application are claimed. Each of the prior mentioned applications is hereby incorporated by reference herein in its entirety.
Protein kinases constitute one of the largest families of human enzymes and regulate many different signaling processes by adding phosphate groups to proteins; particularly tyrosine kinases phosphorylate proteins on the alcohol moiety of tyrosine residues. The tyrosine kinase family includes members that control cell growth, migration, and differentiation. Abnormal kinase activity has been implicated in a variety of human diseases including cancers, autoimmune and inflammatory diseases. Since protein kinases are among the key regulators of cell signaling they provide a means to modulate cellular function with small molecule inhibitors of kinase activity and thus make good drug design targets. In addition to treatment of kinase-mediated disease processes, selective and efficacious inhibitors of kinase activity are also useful for investigation of cell signaling processes and identification of other cellular targets of therapeutic interest.
SYK (Spleen Tyrosine Kinase) is a non-receptor tyrosine kinase that is essential for B-cell activation through BCR signaling. SYK becomes activated upon binding to phosphorylated BCR and thus initiates the early signaling events following BCR activation. Mice deficient in SYK exhibit an early block in B-cell development. Therefore inhibition of SYK enzymatic activity in cells is proposed as a treatment for autoimmune disease through its effects on autoantibody production.
In addition to the role of SYK in BCR signaling and B-cell activation, it also plays a key role in FcεRI mediated mast cell degranulation and eosinophil activation. Thus, SYK is implicated in allergic disorders including asthma. SYK binds to the phosphorylated gamma chain of FcγRI via its SH2 domains and is essential for downstream signaling. SYK deficient mast cells demonstrate defective degranulation, arachidonic acid and cytokine secretion. This also has been shown for pharmacologic agents that inhibit SYK activity in mast cells. Treatment with SYK antisense oligonucleotides inhibits antigen-induced infiltration of eosinophils and neutrophils in an animal model of asthma. SYK deficient eosinophils also show impaired activation in response to FcεR stimulation. Therefore, small molecule inhibitors of SYK will be useful for treatment of allergy-induced inflammatory diseases including asthma.
In view of the numerous conditions that are contemplated to benefit by treatment involving modulation of the SYK pathway it is immediately apparent that new compounds that modulate the SYK pathway and methods of using these compounds should provide substantial therapeutic benefits to a wide variety of patients. Provided herein are novel compounds for use in the therapeutic treatment of auto-immune and inflammatory diseases by targeting the SYK pathway or by inhibition of SYK kinase . . .
R1 is H, C(═O)OR1′, NHC(═O)(CR1″2)nR1′, C(═O)NH(CR1″2)nR1′, or NHR1″;
R1′ is lower alkyl, cycloalkyl, heterocycloalkyl, lower alkyl heterocycloalkyl, O(═O)R1″, OH, or heteroaryl;
each R1″ is independently H or lower alkyl;
R2 is monocyclic or bicyclic unsaturated or partially saturated heteroaryl, optionally substituted with one or more R2′;
each R2′ is independently lower alkyl, lower alkoxy, or phenyl;
X1 is NH or CH; and
X2 is CH or S;
The application provides a method for treating an inflammatory or autoimmune condition comprising administering to a patient in need thereof a therapeutically effective amount of the compound of Formula I.
Certain compounds may exhibit tautomerism. Tautomeric compounds can exist as two or more interconvertable species. Prototropic tautomers result from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium and attempts to isolate an individual tautomers usually produce a mixture whose chemical and physical properties are consistent with a mixture of compounds. The position of the equilibrium is dependent on chemical features within the molecule. For example, in many aliphatic aldehydes and ketones, such as acetaldehyde, the keto form predominates while; in phenols, the enol form predominates. Common prototropic tautomers include keto/enol (—C(═O)—CH-Δ-C(—OH)═CH—), amide/imidic acid (—C(═O)—NH-Δ-C(—OH)═N—) and amidine (—C(═NR)—NH-Δ-C(—NHR)═N—) tautomers. The latter two are particularly common in heteroaryl and heterocyclic rings and the present invention encompasses all tautomeric forms of the compounds.
The definitions described herein may be appended to form chemically-relevant combinations, such as “heteroalkylaryl,” “haloalkylheteroaryl,” “arylalkylheterocyclyl,” “alkylcarbonyl,” “alkoxyalkyl,” and the like. When the term “alkyl” is used as a suffix following another term, as in “phenylalkyl,” or “hydroxyalkyl,” this is intended to refer to an alkyl group, as defined above, being substituted with one to two substituents selected from the other specifically-named group. Thus, for example, “phenylalkyl” refers to an alkyl group having one to two phenyl substituents, and thus includes benzyl, phenylethyl, and biphenyl. An “alkylaminoalkyl” is an alkyl group having one to two alkylamino substituents. “Hydroxyalkyl” includes 2-hydroxyethyl, 2-hydroxypropyl, 1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 2,3-dihydroxybutyl, 2-(hydroxymethyl), 3-hydroxypropyl, and so forth. Accordingly, as used herein, the term “hydroxyalkyl” is used to define a subset of heteroalkyl groups defined below. The term—(ar)alkyl refers to either an unsubstituted alkyl or an aralkyl group. The term (hetero)aryl or (het)aryl refers to either an aryl or a heteroaryl group.
R1 is H, C(═O)OR1′, NHC(═O)(CR1″2)nR1′, C(═O)NH(CR1″2)nR1′, or NHR1′;
R1 is H, C(═O)OR1′, NHC(═O)(CR1″2)nR1′, C(═O)NH(CR1″2)nR1′, or NHR1;
The application provides a compound of Formula I, wherein R2 is bicyclic unsaturated or partially saturated heteroaryl, optionally substituted with one or more R2′.
The application provides a compound of Formula I, wherein R2 is thiazolyl, triazolopyridinyl, benzothiazolyl, benzothiophenyl, benzoimidazolyl, cyclopentathiazolyl, imidazopyridinyl, pyridinyl, tetrahydorbenzothiazolyl, triazolopyridinyl optionally substituted with one or more R2′.
The application provides a compound of Formula I, wherein X1 is NH and X2 is CH.
The application provides a compound of Formula I, wherein X1 is CH and X2 is S.
The application provides a compound of Formula I, wherein X1 is NH, X2 is CH and R2 is bicyclic unsaturated or partially saturated heteroaryl, optionally substituted with one or more R2′.
The application provides a compound of Formula I, wherein X1 is CH, X2 is S, and R2 is bicyclic unsaturated or partially saturated heteroaryl, optionally substituted with one or more R2′.
The application provides a compound of Formula I, wherein R1 is C(═O)NH(CR1″2)nR1′.
The application provides a compound of Formula I, wherein R1 is H.
The application provides a compound of Formula I, wherein R1 is NHC(═O)(CR″2)nR1′.
The application provides a compound of Formula I, wherein R1 is NHR1.
The application provides a compound of Formula I, wherein R1 is C(═O)OR1′.
The application provides a compound of Formula I, wherein R1 is C(═O)NH(CR1″2)nR1′ and R2 is bicyclic unsaturated or partially saturated heteroaryl, optionally substituted with one or more R2′.
The application provides a compound of Formula I, wherein R1 is H and R2 is bicyclic unsaturated or partially saturated heteroaryl, optionally substituted with one or more R2′.
The application provides a compound of Formula I, wherein R1 is NHC(═O)(CR″2)nR1′ and R2 is bicyclic unsaturated or partially saturated heteroaryl, optionally substituted with one or more R2′.
The application provides a compound of Formula I, wherein R1 is NHR1 and R2 is bicyclic unsaturated or partially saturated heteroaryl, optionally substituted with one or more R2′.
The application provides a compound of Formula I, wherein R1 is C(═O)OR1′ and R2 is bicyclic unsaturated or partially saturated heteroaryl, optionally substituted with one or more R2′.
The application provides a compound of Formula I, wherein R2 is monocyclic unsaturated heteroaryl, optionally substituted with one or more R2′.
The application provides a compound of Formula I, wherein R2 is monocyclic unsaturated heteroaryl, optionally substituted with one or more R2′ and R1 is C(═O)NH(CR1″2)nR1′.
2-Phenyl-5-(1H-pyrazol-3-yl)-thiazole;
3-[1,2,4]Triazolo[1,5-a]pyridin-2-yl-1H-pyrazole-4-carboxylic acid ethyl ester;
N-(3-Benzothiazol-2-yl-1H-pyrazol-4-yl)-isobutyramide;
N-(3-Benzothiazol-2-yl-1H-pyrazol-4-yl)-2,2-dimethyl-propionamide;
Cyclopentanecarboxylic acid (3-benzothiazol-2-yl-1H-pyrazol-4-yl)-amide;
Cyclopropanecarboxylic acid (3-benzothiazol-2-yl-1H-pyrazol-4-yl)-amide;
3-(benzo[d]thiazol-2-yl)-N-(1-hydroxy-2-methylpropan-2-yl)-1H-pyrazole-4-carboxamide;
3-(benzo[d]thiazol-2-yl)-N-tert-butyl-1H-pyrazole-4-carboxamide;
N-(1-hydroxy-2-methylpropan-2-yl)-3-(5-methylbenzo[b]thiophen-2-yl)-1H-pyrazole-4-carboxamide;
N-tert-butyl-3-(5-methylbenzo[b]thiophen-2-yl)-1H-pyrazole-4-carboxamide;
N-tert-butyl-3-(6-methoxypyridin-2-yl)-1H-pyrazole-4-carboxamide;
(S)-3-(benzo[d]thiazol-2-yl)-N-(1-hydroxypropan-2-yl)-1H-pyrazole-4-carboxamide;
ethyl 3-(4-phenylthiazol-2-yl)-1H-pyrazole-4-carboxylate;
ethyl 3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-pyrazole-4-carboxylate;
ethyl 3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1H-pyrazole-4-carboxylate;
N-tert-butyl-3-(4-phenylthiazol-2-yl)-1H-pyrazole-4-carboxamide;
N-isopropyl-3-(4-phenylthiazol-2-yl)-1H-pyrazole-4-carboxamide;
N-tert-butyl-3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1H-pyrazole-4-carboxamide;
N-tert-butyl-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-pyrazole-4-carboxamide;
N-cyclopentyl-3-(4-phenylthiazol-2-yl)-1H-pyrazole-4-carboxamide;
3-(4-phenylthiazol-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazole-4-carboxamide;
3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazole-4-carboxamide;
N-(tetrahydro-2H-pyran-4-yl)-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-pyrazole-4-carboxamide;
N-(1-methylpiperidin-4-yl)-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-pyrazole-4-carboxamide;
Tetrahydro-pyran-4-carboxylic acid (3-benzothiazol-2-yl-1H-pyrazol-4-yl)-amide;
Cyclobutanecarboxylic acid (3-benzothiazol-2-yl-1H-pyrazol-4-yl)-amide;
Acetic acid 2-(3-benzothiazol-2-yl-1H-pyrazol-4-ylcarbamoyl)-2-methyl-propyl ester;
N-(3-Benzothiazol-2-yl-1H-pyrazol-4-yl)-3-hydroxy-2,2-dimethyl-propionamide;
4-(5-Methyl-1H-benzoimidazol-2-yl)-thiazole-5-carboxylic acid tert-butylamide;
N-[3-(1H-Benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide;
N-[3-(1H-Benzoimidazol-2-yl)-1H-pyrazol-4-yl]-2,2-dimethyl-propionamide;
Cyclopentanecarboxylic acid [3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-amide;
Tetrahydro-pyran-4-carboxylic acid [3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-amide;
Acetic acid 2-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-ylcarbamoyl]-2-methyl-propyl ester;
N-[3-(1H-Benzoimidazol-2-yl)-1H-pyrazol-4-yl]-3-hydroxy-2,2-dimethyl-propionamide;
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid isopropylamide;—12—case 31176
Inhibitors of SYK
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid tert-butylamide;
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid (tetrahydro-pyran-4-yl)-amide;
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid (2-hydroxy-1,1-dimethyl-ethyl)-amide;
4-(4-methyl-1H-benzo[d]imidazol-2-yl)-N-(pyridin-3-yl)thiazol-5-amine;
4-(1H-imidazo[4,5-c]pyridin-2-yl)-N-(pyridin-3-yl)thiazol-5-amine;
4-(5-methyl-1H-benzo[d]imidazol-2-yl)-N-tert-pentylthiazol-5-amine;
N-cyclopentyl-4-(5-methyl-1H-benzo[d]imidazol-2-yl)thiazol-5-amine; and
N-isobutyl-4-(5-methyl-1H-benzo[d]imidazol-2-yl)thiazol-5-amine.
The application provides the above method, further comprising administering an additional therapeutic agent selected from a chemotherapeutic or anti-proliferative agent, an anti-inflammatory agent, an immunomodulatory or immunosuppressive agent, a neurotrophic factor, an agent for treating cardiovascular disease, an agent for treating diabetes, or an agent for treating immunodeficiency disorders.
The application provides a method for treating an immune disorder including lupus, multiple sclerosis, rheumatoid arthritis, psoriasis, Type I diabetes, complications from organ transplants, xeno transplantation, diabetes, cancer, asthma, atopic dermatitis, autoimmune thyroid disorders, ulcerative colitis, Crohn's disease, Alzheimer's disease, and Leukemia, comprising administering to a patient in need thereof a therapeutically effective amount of the compound of Formula I.
The application provides a method for treating an inflammatory condition comprising co-administering to a patient in need thereof a therapeutically effective amount of an anti-inflammatory compound in combination with the compound of Formula I.
The application provides a method for treating an immune disorder comprising co-administering to a patient in need thereof a therapeutically effective amount of an immunosuppressant compound in combination with the compound of Formula I.
The application provides the above pharmaceutical composition, further comprising an additional therapeutic agent selected from a chemotherapeutic or anti-proliferative agent, an anti-inflammatory agent, an immunomodulatory or immunosuppressive agent, a neurotrophic factor, an agent for treating cardiovascular disease, an agent for treating diabetes, and an agent for treating immunodeficiency disorders.
The application provides the use of the compound of formula I for the manufacture of a medicament useful for the treatment of disorders associated with Syk.
The application provides the use of the compound of formula I for the manufacture of a medicament useful for the treatment of rheumatoid arthritis.
A compound, method, or composition as described herein.
In general, the nomenclature used in this Application is based on AUTONOM™ v.4.0, a Beilstein Institute computerized system or Struct=Name, a CambridgeSoft® application, for the generation of IUPAC systematic nomenclature. If there is a discrepancy between a depicted structure and a name given that structure, the depicted structure is to be accorded more weight. In addition, if the stereochemistry of a structure or a portion of a structure is not indicated with, for example, bold or dashed lines, the structure or portion of the structure is to be interpreted as encompassing all stereoisomers of it.
TABLE I Com- pound Nomenclature Structure
I-1 2-Phenyl-5-(1H-pyrazol-3- yl)-thiazole I-2 3-[1,2,4]Triazolo[1,5- a]pyridin-2-yl-1H-pyrazole- 4-carboxylic acid ethyl ester I-3 N-(3-Benzothiazol-2-yl-1H- pyrazol-4-yl)-isobutyramide I-4 N-(3-Benzothiazol-2-yl-1H- pyrazol-4-yl)-2,2-dimethyl- propionamide I-5 Cyclopenanecarboxylic acid (3-benzothiazol-2-yl-1H- pyrazol-4-yl)-amide I-6 Cyclopropanecarboxylic acid (3-benzothiazol-2-yl-1H- pyrazol-4-yl)-amide I-7 3-(benzo[d]thiazol-2-yl)-N- (1-hydroxy-2-methylpropan- 2-yl)-1H-pyrazole-4- carboxamide I-8 3-(benzo[d]thiazol-2-yl)-N- tert-butyl-1H-pyrazole-4- carboxamide I-9 N-(1-hydroxy-2- methylpropan-2-yl)-3-(5- methylbenzo[b]thiophen-2- yl)-1H-pyrazole-4- carboxamide I-10 N-tert-butyl-3-(5- methylbenzo[b]thiophen-2- yl)-1H-pyrazole-4- carboxamide I-11 N-tert-butyl-3-(6- methoxypyridin-2-yl)-1H- pyrazole-4-carboxamide I-12 (S)-3-(benzo[d]thiazol-2-yl)- N-(1-hydroxypropan-2-yl)- 1H-pyrazole-4-carboxamide I-13 ethyl 3-(4-phenylthiazol-2- yl)-1H-pyrazole-4- carboxylate I-14 ethyl 3-(4,5,6,7- tetrahydrobenzo[d]thiazol-2- yl)-1H-pyrazole-4- carboxylate I-15 ethyl 3-(5,6-dihydro-4H- cyclopenta[d]thiazol-2-yl)- 1H-pyrazole-4-carboxylate I-16 N-tert-butyl-3-(4- phenylthiazol-2-yl)-1H- pyrazole-4-carboxamide I-17 N-isopropyl-3-(4- phenylthiazol-2-yl)-1H- pyrazole-4-carboxamide I-18 N-tert-butyl-3-(5,6-dihydro- 4H-cyclopenta[d]thiazol-2- yl)-1H-pyrazole-4- carboxamide I-19 N-tert-butyl-3-(4,5,6,7- tetrahydrobenzo[d]thiazol-2- yl)-1H-pyrazole-4- carboxamide I-20 N-cyclopentyl-3-(4- phenylthiazol-2-yl)-1H- pyrazole-4-carboxamide I-21 3-(4-phenylthiazol-2-yl)-N- (tetrahydro-2H-pyran-4-yl)- 1H-pyrazole-4-carboxamide I-22 3-(5,6-dihydro-4H- cyclopenta[d]thiazol-2-yl)- N-(tetrahydro-2H-pyran-4- yl)-1H-pyrazole-4- carboxamide I-23 N-(tetrahydro-2H-pyran-4- yl)-3-(4,5,6,7- tetrahydrobenzo[d]thiazol-2- yl)-1H-pyrazole-4- carboxamide I-24 N-(1-methylpiperidin-4-yl)- 3-(4,5,6,7- tetrahydrobenzo[d]thiazol-2- yl)-1H-pyrazole-4- carboxamide I-25 Tetrahydro-pyran-4- carboxylic acid (3- benzothiazol-2-yl-1H- pyrazol-4-yl)-amide I-26 Cyclobutanecarboxylic acid (3-benzothiazol-2-yl-1H- pyrazol-4-yl)-amide I-27 Acetic acid 2-(3- benzothiazol-2-yl-1H- pyrazol-4-ylcarbamoyl)-2- methyl-propyl ester I-28 N-(3-Benzothiazol-2-yl-1H- pyrazol-4-yl)-3-hydroxy-2,2- dimethyl-propionamide I-29 4-(5-Methyl-1H- benzoimidazol-2-yl)- thiazole-5-carboxylic acid tert-butylamide I-30 N-[3-(1H-Benzoimidazol-2- yl)-1H-pyrazoll-4-yl]- isobutyramide I-31 N-[3-(1H-Benzoimidazol-2- yl)-1H-pyrazol-4-yl]-2,2- dimethyl-propionamide I-32 Cyclopentanecarboxylic acid [3-(1H-benzoimidazol-2-yl)- 1H-pyrazol-4-yl]-amide I-33 Tetrahydro-pyran-4- carboxylic acid [3-(1H- benzoimidazol-2-yl)-1H- pyrazol-4-yl]-amide I-34 Acetic acid 2-[3-(1H- benzoimidazol-2-yl)-1H- pyrazol-4-ylcarbamoyl]-2- methyl-propyl ester I-35 N-[3-(1H-Benzoimidazol-2- yl)-1H-pyrazol-4-yl]-3- hydroxy-2,2-dimethyl- propionamide I-36 3-(1H-Benzoimidazol-2-yl)- 1H-pyrazole-4-carboxylic acid isopropylamide I-37 3-(1H-Benzoimidazol-2-yl)- 1H-pyrazole-4-carboxylic acid tert-butylamide I-38 3-(1H-Benzoimidazol-2-yl)- 1H-pyrazole-4-carboxylic acid (tetrahydro-pyran-4-yl)- amide I-39 3-(1H-Benzoimidazol-2-yl)- 1H-pyrazole-4-carboxylic acid (2-hydroxy-1,1- dimethyl-ethyl)-amide I-40 4-(4-methyl-1H- benzo[d]imidazol-2-yl)-N- (pyridin-3-yl)thiazol-5- amine I-41 4-(1H-imidazo[4,5- c]pyridin-2-yl)-N-(pyridin-3- yl)thiazol-5-amide I-42 4-(5-methyl-1H- benzo[d]imidazol-2-yl)-N- tert-pentylthiazol-5-amine I-43 N-cyclopentyl-4-(5-methyl- 1H-benzo[d]imidazol-2- yl)thiazol-5-amine I-44 N-isobutyl-4-(5-methyl-1H- benzo[d]imidazol-2- yl)thiazol-5-amine
The term “therapeutically effective amount” as used herein means an amount required to reduce symptoms of the disease in an individual. The dose will be adjusted to the individual requirements in each particular case. That dosage can vary within wide limits depending upon numerous factors such as the severity of the disease to be treated, the age and general health condition of the patient, other medicaments with which the patient is being treated, the route and form of administration and the preferences and experience of the medical practitioner involved. For oral administration, a daily dosage of between about 0.01 and about 1000 mg/kg body weight per day should be appropriate in monotherapy and/or in combination therapy. A preferred daily dosage is between about 0.1 and about 500 mg/kg body weight, more preferred 0.1 and about 100 mg/kg body weight and most preferred 1.0 and about 10 mg/kg body weight per day.
Thus, for administration to a 70 kg person, the dosage range would be about 7 mg to 0.7 g per day. The daily dosage can be administered as a single dosage or in divided dosages, typically between 1 and 5 dosages per day. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect for the individual patient is reached. One of ordinary skill in treating diseases described herein will be able, without undue experimentation and in reliance on personal knowledge, experience and the disclosures of this application, to ascertain a therapeutically effective amount of the compounds of the present invention for a given disease and patient.
Active ingredient 0.25 g Sodium Chloride qs to make isotonic Water for injection 100 ml
Several aqueous suspensions containing from about 0.025-0.5 percent active compound are prepared as nasal spray formulations. The formulations optionally contain inactive ingredients such as, for example, microcrystalline cellulose, sodium carboxymethylcellulose, dextrose, and the like. Hydrochloric acid may be added to adjust pH. The nasal spray formulations may be delivered via a nasal spray metered pump typically delivering about 50-100 microliters of formulation per actuation. A typical dosing schedule is 2-4 sprays every 4-12 h.
The compounds described herein are kinase inhibitors, in particular SYK inhibitors. These inhibitors can be useful for treating one or more diseases responsive to kinase inhibition, including diseases responsive to SYK inhibition and/or inhibition of B-cell proliferation, in mammals. Without wishing to be bound to any particular theory, it is believed that the interaction of the compounds of the invention with SYK results in the inhibition of SYK activity and thus in the pharmaceutical utility of these compounds. Accordingly, the invention includes a method of treating a mammal, for instance a human, having a disease responsive to inhibition of SYK activity, and/or inhibiting B-cell proliferation, comprising administrating to the mammal having such a disease, an effective amount of at least one chemical entity provided herein. An effective concentration may be ascertained experimentally, for example by assaying blood concentration of the compound, or theoretically, by calculating bioavailability. Other kinases that may be affected in addition to SYK include, but are not limited to, other tyrosine kinases and serine/threonine kinases.
Commonly used abbreviations include: acetyl (Ac), azo-bis-isobutyrylnitrile (AIBN), atmospheres (Atm), 9-borabicyclo[3.3.1]nonane (9-BBN or BBN), 2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (BINAP), tert-butoxycarbonyl (Boc), di-tert-butyl pyrocarbonate or boc anhydride (BOC2O), benzyl (Bn), butyl (Bu), Chemical Abstracts Registration Number (CASRN), benzyloxycarbonyl (CBZ or Z), carbonyl diimidazole (CDI), 1,4-diazabicyclo[2.2.2]octane (DABCO), diethylaminosulfur trifluoride (DAST), dibenzylideneacetone (dba), 1,5-diazabicyclo[4.3.0]non-5-ene (DBN), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), N,N′-dicyclohexylcarbodiimide (DCC), 1,2-dichloroethane (DCE), dichloromethane (DCM), 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), diethyl azodicarboxylate (DEAD), di-iso-propylazodicarboxylate (DIAD), di-iso-butylaluminumhydride (DIBAL or DIBAL-H), di-iso-propylethylamine (DIPEA), N,N-dimethyl acetamide (DMA), 4-N,N-dimethylaminopyridine (DMAP), N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,1′-bis-(diphenylphosphino)ethane (dppe), 1,1′-bis-(diphenylphosphino)ferrocene (dppf), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ), ethyl (Et), ethyl acetate (EtOAc), ethanol (EtOH), 2-ethoxy-2H-quinoline-1-carboxylic acid ethyl ester (EEDQ), diethyl ether (Et2O), ethyl isopropyl ether (EtOiPr), O-(7-azabenzotriazole-1-yl)-N, N,N′N′-tetramethyluronium hexafluorophosphate acetic acid (HATU), acetic acid (HOAc), 1-N-hydroxybenzotriazole (HOBt), high pressure liquid chromatography (HPLC), iso-propanol (IPA), isopropylmagnesium chloride (iPrMgCl), hexamethyl disilazane (HMDS), liquid chromatography mass spectrometry (LCMS), lithium hexamethyl disilazane (LiHMDS), meta-chloroperoxybenzoic acid (m-CPBA), methanol (MeOH), melting point (mp), MeSO2— (mesyl or Ms), methyl (Me), acetonitrile (MeCN), m-chloroperbenzoic acid (MCPBA), mass spectrum (ms), methyl t-butyl ether (MTBE), methyl tetrahydrofuran (MeTHF), N-bromosuccinimide (NBS), n-Butyllithium (nBuLi), N-carboxyanhydride (NCA), N-chlorosuccinimide (NCS), N-methylmorpholine (NMM), N-methylpyrrolidone (NMP), pyridinium chlorochromate (PCC), Dichloro-((bis-diphenylphosphino)ferrocenyl) palladium(II) (Pd(dppf)Cl2), palladium(II) acetate (Pd(OAc)2), tris(dibenzylideneacetone)dipalladium(0) (Pd2(dba)3), pyridinium dichromate (PDC), phenyl (Ph), propyl (Pr), iso-propyl (i-Pr), pounds per square inch (psi), pyridine (pyr), 1,2,3,4,5-Pentaphenyl-1′(di-tert-butylphosphino)ferrocene (Q-Phos), room temperature (ambient temperature, rt or RT), sec-Butyllithium (sBuLi), tert-butyldimethylsilyl or t-BuMe2Si (TBDMS), tetra-n-butylammonium fluoride (TBAF), triethylamine (TEA or Et3N), 2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO), triflate or CF3SO2-(Tf), trifluoroacetic acid (TFA), 1,1′-bis-2,2,6,6-tetramethylheptane-2,6-dione (TMHD), O-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU), thin layer chromatography (TLC), tetrahydrofuran (THF), trimethylsilyl or Me3Si (TMS), p-toluenesulfonic acid monohydrate (TsOH or pTsOH), 4-Me-C6H4SO2— or tosyl (Ts), and N-urethane-N-carboxyanhydride (UNCA). Conventional nomenclature including the prefixes normal (n), iso (i-), secondary (sec-), tertiary (tert-) and neo have their customary meaning when used with an alkyl moiety. (J. Rigaudy and D. P. Klesney, Nomenclature in Organic Chemistry, IUPAC 1979 Pergamon Press, Oxford.).
Unless otherwise stated, all temperatures including melting points (i.e., MP) are in degrees Celsius (° C.). It should be appreciated that the reaction which produces the indicated and/or the desired product may not necessarily result directly from the combination of two reagents which were initially added, i.e., there may be one or more intermediates which are produced in the mixture which ultimately leads to the formation of the indicated and/or the desired product. The preceding abbreviations may be used in the Preparations and Examples. All names were generated using Autonom or ChemDraw.
Ethyl 3-([1,2,4]triazolo[1,5-a]pyridin-2-yl)-1H-pyrazole-4-carboxylate
Ethyl [1,2,4]triazolo[1,5-a]pyridine-2-carboxylate
To solution of ethyl 2-ethoxy-2-iminoacete (1.91 g, 13.2 mmol) in ethanol (66.1 mL) was added 1,2-diaminopyridinium iodide (2.35 g, 9.91 mmol) followed by potassium hydroxide (556 mg, 9.91 mmol) and stirred at r.t. for 16 h. Water was then added, the mixture extracted with EtOAc, the organic layer was separated, and concentrated in vacuo. Purification by chromatography (silica, 60-100% ethyl acetate in hexanes) gave ethyl [1,2,4]triazolo[1,5-a]pyridine-2-carboxylate (570 mg, 30%). 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 8.67 (dt, J=6.9, 1.2 Hz, 1 H), 7.86 (dt, J=9.1, 1.2 Hz, 1 H), 7.62 (m, 1 H), 7.17 (td, J=7.0, 1.3 Hz, 1 H), 4.56 (q, J=7.2 Hz, 2 H), 1.48 (t, J=7.1 Hz, 3 H).
Ethyl 3-([1,2,4]triazolo[1,5-a]pyridin-2-yl)-3-oxopropanoate
To a mixture of ethyl [1,2,4]triazolo[1,5-a]pyridine-2-carboxylate (590 mg, 3.09 mmol) and ethyl acetate (1.81 mL, 18.5 mmol) in THF (6 mL) at −50° C. was quickly added lithium bis(trimethylsilyl)amide (1 M in toluene, 9.26 mL, 9.26 mmol). The mixture was stirred for 30 min, and then quenched with acetic acid, washed with water, sodium bicarbonate, and brine. Purification by chromatography (silica, 50-100% ethyl acetate in hexanes) gave ethyl 3-([1,2,4]triazolo[1,5-a]pyridin-2-yl)-3-oxopropanoate (350 mg, 49%) as a clear oil.
(E)-Ethyl 2-([1,2,4]triazolo[1,5-a]pyridine-2-carbonyl)-3-(dimethylamino)acrylate
To a solution of ethyl 3-([1,2,4]triazolo[1,5-a]pyridin-2-yl)-3-oxopropanoate (350 mg, 1.5 mmol) in ethanol (4.3 mL) was added dimethylformamide dimethyl acetal (1.97 mg, 1.65 mmol) and the mixture heated to 80° C. for 3 h. The mixture was concentrated in vacuo, adsorbed onto silica, and purified by chromatography (silica, 1-9% methanol in dichloromethane) to give (E)-ethyl 2-([1,2,4]triazolo[1,5-a]pyridine-2-carbonyl)-3-(dimethylamino)acrylate (250 mg, 58%). 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 8.61 (d, J=6.9 Hz, 1 H), 7.81 (s, 1 H), 7.79 (d, J=8.9 Hz, 1 H), 7.55 (t, J=7.7 Hz, 1 H), 7.08 (t, J=6.9 Hz, 1 H), 4.01 (q, J=7.3 Hz, 2 H), 3.31 (br. s, 3 H), 2.93 (br. s, 3 H), 0.88 (t, J=7.1 Hz, 3 H).
To a solution of (E)-ethyl 2-([1,2,4]triazolo[1,5-a]pyridine-2-carbonyl)-3-(dimethylamino)acrylate (250 mg, 0.867 mmol) in ethanol (8.7 mL) was added hydrazine hydrate (47.3 mg, 0.954 mmol) and the resultant mixture stirred at r.t. for 2 h. The mixture was concentrated in vacuo then purified by chromatography (silica, 1-10% methanol in dichloromethane) to give ethyl 3-([1,2,4]triazolo[1,5-a]pyridin-2-yl)-1H-pyrazole-4-carboxylate (170 mg, 76%) as an off-white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 8.76 (d, J=7.0 Hz, 1 H), 8.24 (s, 1 H), 7.90 (d, J=9.1 Hz, 1 H), 7.61 (t, J=7.9 Hz, 1 H), 7.12 (t, J=6.9 Hz, 1 H), 4.40 (q, J=7.2 Hz, 2 H), 1.39 (t, J=7.2 Hz, 3 H); MS (EI/CI) m/z: 257.9 [M+H].
N-(3-Benzothiazol-2-yl-1H-pyrazol-4-yl)-isobutyramide
2-(4-Nitro-1H-pyrazol-3-yl)-benzothiazole
To a solution of 4-nitro-1H-pyrazole-3-carboxylic acid (5 g, 31.8 mmol) in THF (40 mL) was added DMF (0.25 mL, 3.19 mmol) and oxalyl chloride (4.1 mL, 47.8 mmol). The mixture was stirred at room temperature for 1 h and then concentrated under reduced pressure. The crude material was redissolved in NMP (40 mL) and 2-amino-benzenethiol (4.0 mL, 31.8 mmol) was added. This mixture was heated at 100° C. for 1 h, at which point water (100 mL) was added and the aqueous phase extracted with ethyl acetate (3×25 mL). The combined organic layers were dried (Na2SO4), concentrated under reduced pressure, and purified by chromatography (silica, EtOAc/hexanes) to give 2-(4-nitro-1H-pyrazol-3-yl)-benzothiazole as a light yellow solid (4.8 g, 61%). MS (EI/CI) m/z: 245.0 [M−H].
3-Benzothiazol-2-yl-1H-pyrazol-4-ylamine
A mixture of 2-(4-nitro-1H-pyrazol-3-yl)-benzothiazole (1.2 g, 4.87 mmol) and 10% Pd—C(50% moist) (1.7 g) in DMF (10 mL) was stirred vigorously under an atmosphere of hydrogen (balloon pressure) at room temperature for 20 h. The reaction mixture was passed through celite and the filtrate concentrated under reduced pressure. The remaining residue was partitioned between ethyl acetate and water, the organic layer separated, dried over anhydrous Na2SO4, and concentrated under reduced pressure to give 3-benzothiazol-2-yl-1H-pyrazol-4-ylamine as brown solid (0.76 g, 72%), which was used directly in the next step without further purification. MS (EI/CI) m/z: 217.2 [M+H].
N-(3-benzothiazol-2-yl-1H-pyrazol-4-yl)-isobutyramide (148.0 mg, 56%) was synthesized as light yellow solid from 3-benzothiazol-2-yl-1H-pyrazol-4-ylamine (200 mg, 0.926 mmol) and isobutyryl chloride (0.32 mL, 3.06 mmol) following the procedure described for N-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide (Example 29). MS (EI/CI) m/z: 287.1 [M+H].
N-(3-Benzothiazol-2-yl-1H-pyrazol-4-yl)-2,2-dimethyl-propionamide
N-(3-benzothiazol-2-yl-1H-pyrazol-4-yl)-2,2-dimethyl-propionamide (115 mg, 41%) was synthesized as off white solid from 3-benzothiazol-2-yl-1H-pyrazol-4-ylamine (200 mg, 0.926 mmol) and pivaloyl chloride (0.38 mL, 3.056 mmol) following the procedure described for N-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide (Example 29). MS (EI/CI) m/z: 301.2 [M+H].
Cyclopentanecarboxylic acid (3-benzothiazol-2-yl-1H-pyrazol-4-yl)-amide
Cyclopentanecarboxylic acid (3-benzothiazol-2-yl-1H-pyrazol-4-yl)-amide (124.0 mg, 43%) was synthesized as off white solid from 3-benzothiazol-2-yl-1H-pyrazol-4-ylamine (200 mg, 0.926 mmol) and cyclopentanecarbonyl chloride (0.37 mL, 3.06 mmol) following the procedure described for N-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide (Example 29). MS (EI/CI) m/z: 313.1 [M+H].
Cyclopropanecarboxylic acid (3-benzothiazol-2-yl-1H-pyrazol-4-yl)-amide
Cyclopropanecarboxylic acid (3-benzothiazol-2-yl-1H-pyrazol-4-yl)-amide (131 mg, 50%) was synthesized as an off white solid from 3-benzothiazol-2-yl-1H-pyrazol-4-ylamine (200 mg, 0.93 mmol) and cyclopropanecarbonyl chloride (0.28 mL, 3.06 mmol) following the procedure described for N-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide (Example 29). MS (EI/CI) m/z: 285.2 [M+H].
3-(Benzo[d]thiazol-2-yl)-N-(1-hydroxy-2-methylpropan-2-yl)-1H-pyrazole-4-carboxamide
To a solution of ethyl 3-amino-1H-pyrazole-4-carboxylate (1 g, 6.45 mmol) in diiodomethane (25.9 g, 7.8 mL, 97 mmol) at −10° C. was added isoamyl nitrite (3.4 g, 3.91 mL, 29.0 mmol) dropwise over 5 min. The reaction was heated to 100° C. for 2 h, then cooled and directly purified by chromatography (silica, 0-5% methanol in dichloromethane) to give ethyl 3-iodo-1H-pyrazole-4-carboxylate (1.20 g, 4.51 mmol, 70%) as an orange solid. MS (EI/CI) m/z: 266.6 [M+H].
Ethyl 3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate
To a 0° C. solution of ethyl 3-iodo-1H-pyrazole-4-carboxylate (1.2 g, 4.51 mmol) in THF (22.4 mL) was added sodium hydride (361 mg, 9.02 mmol) and the mixture stirred for 30 min. (2-(Chloromethoxy)ethyl)trimethylsilane (902 mg, 5.41 mmol) was then added and the reaction mixture was stirred at r.t. for 16 h, quenched with sodium bicarbonate (aqueous, saturated) and extracted into ether. The combined organic solvents were concentrated in vacuo then purified by chromatography (silica, 0-15% ethyl acetate in hexanes) to give ethyl 3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate (366 mg, 924 μmol, 21%) as a colorless liquid. 1H NMR (400 MHz,CHLOROFORM-d) δ ppm 7.99 (s, 1 H), 5.42 (s, 2 H), 4.34 (q, J=7.0 Hz, 2 H), 3.61 (t, J=8.5 Hz, 2 H), 1.38 (t, J=7.2 Hz, 3 H), 0.92 (t, J=8.1 Hz, 2 H), 0.01 (s, 9 H).
Ethyl 3-(benzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate
Ethyl 3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate (239 mg, 603 μmol), 2-(tributylstannyl)benzo[d]thiazole (384 mg, 905 μmol), tetrakis(triphenylphosphine)palladium (0) (70 mg, 60 μmol) and copper (I) iodide (17 mg, 91 μmol) in dioxane (2.4 mL) and DMF (398 μL) were heated to 95° C. for 16 h. After which the mixture was cooled, concentrated in vacuo, and purified by chromatography (silica, 10-40% ethyl acetate in hexanes) to give ethyl 3-(benzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate (100 mg, 248 μmol, 41%) as an off-white solid. MS (EI/CI) m/z: 404.4 [M+H].
3-(Benzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid
To a solution of ethyl 3-(benzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate (144 mg, 357 μmol) in dioxane (5 mL) was added sodium hydroxide (1N, 1.78 mL, 1.78 mmol). The mixture was stirred at r.t. for 16 h and then quenched with 10% HCl (˜pH 6), diluted with ethyl acetate and washed with water (2×). The organic layer was dried (MgSO4), filtered, and concentrated in vacuo to give 3-(benzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (115 mg, 306 μmol, 86%) as an off white solid. MS (EI/CI) m/z: 376.1 [M+H].
3-(Benzo[d]thiazol-2-yl)-N-(1-hydroxy-2-methylpropan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide
3-(Benzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (55 mg, 146 μmol), 2-amino-2-methylpropan-1-ol (26.1 mg, 293 μmol), HATU (66.8 mg, 176 μmol) and DIPEA (56.8 mg, 77 μL, 439 μmol) in DMF (1 mL) were stirred at r.t. for 16 h. The mixture was quenched with 10% citric acid, diluted with ethyl acetate, and the phases separated. The organic phase was washed with sodium bicarbonate and brine, then purified by chromatography (silica, 25-55% ethyl acetate in hexanes) to give 3-(benzo[d]thiazol-2-yl)-N-(1-hydroxy-2-methylpropan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (50 mg, 112 μmol, 76%) as a white solid. MS (EI/CI) m/z: 477.1 [M+H].
To a solution of 3-(benzo[d]thiazol-2-yl)-N-(1-hydroxy-2-methylpropan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (50 mg, 112 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (255 mg, 172 μL, 2.24 mmol). The mixture was stirred at r.t. for 16 h, and then concentrated in vacuo. To this residue was added dichloromethane (2 mL), methanol (1 mL) and ammonium hydroxide (300 μL). This mixture was stirred for an additional 2 h at r.t. and then concentrated in vacuo. The resulting residue was triturated with water, and filtered. This solid was purified by chromatography (silica, 3-7% methanol in dichloromethane) to give 3-(benzo[d]thiazol-2-yl)-N-(1-hydroxy-2-methylpropan-2-yl)-1H-pyrazole-4-carboxamide (21 mg, 66.4 μmol, 59%) as a white solid. 1H NMR (400 MHz, DMSO-d) δ ppm 10.61 (s, 1 H), 8.40 (s, 1 H), 8.19 (d, J=8.2 Hz, 1 H), 8.08 (d, J=8.0 Hz, 1 H), 7.62 (t, J=7.4 Hz, 1 H), 7.55 (t, J=7.5 Hz, 1 H), 5.02 (t, J=5.6 Hz, 1 H), 3.65 (d, J=8.8 Hz, 2 H), 1.46 (s, 6 H); MS (EI/CI) m/z: 317.0 [M+H].
3-(Benzo[d]thiazol-2-yl)-N-tert-butyl-1H-pyrazole-4-carboxamide
3-(Benzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (55 mg, 146 μmol), 2-methylpropan-2-amine (42.8 mg, 61.6 μL, 586 μmol), N1-((ethylimino)methylene)-N3,N3-dimethylpropane-1,3-diamine hydrochloride (64.6 mg, 337 μmol) and HOBT (51.6 mg, 337 μmol) were dissolved in DMF (1 mL) and stirred at r.t. for 16 h. The reaction mixture was quenched with 10% citric acid, diluted with ethyl acetate, and the phases separated. The organic phase was washed with sodium bicarbonate and brine, dried (MgSO4) and evaporated. Purification by chromatography (silica, 5-30% ethyl acetate in hexanes) gave 3-(benzo[d]thiazol-2-yl)-N-tert-butyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (12 mg, 27.9 μmol, 19%). 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 10.90 (s, 1 H), 8.37 (s, 1 H), 7.98 (d, J=7.0 Hz, 1 H), 7.94 (d, J=7.6 Hz, 1 H), 7.52 (t, J=7.0 Hz, 1 H), 7.46 (t, J=7.6 Hz, 1 H), 5.48 (s, 2 H), 3.65 (t, J=8.3 Hz, 2 H), 1.61 (s, 9 H), 0.96 (t, J=8.6 Hz, 2 H), 0.01 (s, 9 H).
To a solution of 3-(benzo[d]thiazol-2-yl)-N-tert-butyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (12 mg, 27.9 μmol) in dichloromethane (0.4 mL) was added trifluoroacetic acid (63.5 mg, 42.9 μL, 557 μmol). The mixture was stirred at r.t. for 16 h then concentrated in vacuo. Dichloromethane (0.5 mL), methanol (0.25 mL) and ammonium hydroxide (0.1 mL) were then added and the mixture was stirred at r.t for an additional 2 h. The mixture was concentrated in vacuo then purified by chromatography (silica, 2.5-5% methanol in dichloromethane) to give 3-(benzo[d]thiazol-2-yl)-N-tert-butyl-1H-pyrazole-4-carboxamide (6 mg, 20.0 μmol, 72%) as a white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 11.07 (s, 1 H), 8.20 (s, 1 H), 7.94 (d, J=8.4 Hz, 1 H), 7.91 (d, J=7.4 Hz, 1 H), 7.48 (t, J=7.1 Hz, 1 H), 7.42 (t, J=7.4 Hz, 1 H), 1.56 (s, 9 H); MS (EI/CI) m/z: 300.8 [M+H].
N-(1-Hydroxy-2-methylpropan-2-yl)-3-(5-methylbenzo[b]thiophen-2-yl)-1H-pyrazole-4-carboxamide
Ethyl 3-(5-methylbenzo[b]thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate
Ethyl 3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate (366 mg, 924 μmol), 5-methylbenzo[b]thiophen-2-ylboronic acid (231 mg, 1.2 mmol), tetrakis(triphenylphosphine)palladium (0) (107 mg, 92.4 μmol) and potassium carbonate (383 mg, 2.77 mmol) in dioxane (16.4 mL) and water (4.1 mL) were heated to 90° C. for 16 h under nitrogen. The mixture was cooled and then concentrated in vacuo. Purification by chromatography (silica, 5-25% ethyl acetate in hexanes) gave ethyl 3-(5-methylbenzo[b]thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate (325 mg, 624 μmol, 68%) as an off-white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 8.38 (s, 1 H), 8.16 (s, 1 H), 7.70 (d, J=8.5 Hz, 1 H), 7.61 (s, 1 H), 7.16 (dd, J=8.0, 1.1 Hz, 1 H), 5.45 (s, 2 H), 4.35 (q, J=7.0 Hz, 2 H), 3.68 (t, J=8.3 Hz, 2 H), 2.45 (s, 3 H), 1.39 (t, J=6.8 Hz, 3 H), 0.95 (t, J=8.2 Hz, 2 H), 0.01 (s, 9 H).
3-(5-Methylbenzo[b]thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid
To a solution of ethyl 3-(5-methylbenzo[b]thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate (325 mg, 780 μmol) in dioxane (11.1 mL) was added sodium hydroxide (1N, 3.9 mL, 3.9 mmol). The mixture was stirred at r.t. for 16 h, then quenched with 10% HCl (˜pH 6) and diluted with ethyl acetate and water. The organic phase was separated washed with water (2×), then concentrated in vacuo. Purification by chromatography (silica, 20-50% ethyl acetate in hexanes) gave 3-(5-methylbenzo[b]thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (185 mg, 476 μmol, 61%) as a white solid. MS (EI/CI) m/z: 389.0 [M+H].
N-(1-Hydroxy-2-methylpropan-2-yl)-3-(5-methylbenzo[b]thiophen-2-yl)-1-((2 (trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide
3-(5-Methylbenzo[b]thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (90 mg, 232 μmol, 2-amino-2-methylpropan-1-ol (41.3 mg, 463 μmol), HATU (106 mg, 278 μmol) and DIPEA (89.8 mg, 121 μL, 695 μmol) in DMF (1.5 mL) were stirred at r.t. for 16 h. The reaction mixture was then quenched with 10% citric acid and diluted with ethyl acetate. The organic phase was separated, washed with sodium bicarbonate and brine, concentrated in vacuo, and then purified by chromatography (silica, 25-90% ethyl acetate in hexanes) to give N-(1-hydroxy-2-methylpropan-2-yl)-3-(5-methylbenzo[b]thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (83 mg, 181 μmol, 78%) as a white solid. MS (EI/CI) m/z: 460.1 [M+H].
To a solution of N-(1-hydroxy-2-methylpropan-2-yl)-3-(5-methylbenzo[b]thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (83 mg, 181 μmol) in dichloromethane (2.8 mL) was added trifluoroacetic acid (412 mg, 278 μL, 3.61 mmol). The mixture was stirred at r.t. for 16 h, then concentrated in vacuo. The residue obtained was redissolved in dichloromethane (2.8 mL), methanol (1.4 mL) and ammonium hydroxide (350 μL) and stirred at r.t. for 1 h. This mixture was concentrated in vacuo and the residue purified by chromatography (silica, 60-100% ethyl acetate in hexanes to give N-(1-hydroxy-2-methylpropan-2-yl)-3-(5-methylbenzo[b]thiophen-2-yl)-1H-pyrazole-4-carboxamide (17 mg, 51.6 μmol, 29%) as a white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.98 (s, 1 H), 7.67 (d, J=8.0 Hz, 1 H), 7.59 (s, 1 H), 7.53 (s, 1 H), 7.16 (d, J=8.4 Hz, 1 H), 6.07 (s, 1 H), 3.58 (s, 2 H), 2.41 (s, 3 H), 1.18 (s, 6 H).; MS (EI/CI) m/z: 329.9 [M+H].
N-tert-Butyl-3-(5-methylbenzo[b]thiophen-2-yl)-1H-pyrazole-4-carboxamide
N-tert-Butyl-3-(5-methylbenzo[b]thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide
3-(5-Methylbenzo[b]thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (90 mg, 232 μmol), 2-methylpropan-2-amine (67.8 mg, 97.4 μl, 927 μmol), N1-((ethylimino)methylene)-N3,N3-dimethylpropane-1,3-diamine hydrochloride (102 mg, 533 μmol) and HOBT (81.6 mg, 533 μmol) were dissolved in DMF (1 mL) and stirred at r.t. for 16 h. The reaction mixture was quenched with 10% citric acid and then diluted with ethyl acetate. The organic phase was separated and washed with sodium bicarbonate and brine. The organic layer was collected, concentrated in vacuo, and purified by chromatography (silica, 5-25% ethyl acetate in hexanes) to give N-tert-butyl-3-(5-methylbenzo[b]thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (72 mg, 162 μmol, 70%) as a colorless oil. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 8.03 (s, 1 H), 7.73 (d, J=8.6 Hz, 1 H), 7.66 (s, 1 H), 7.59 (s, 1 H), 7.20 (d, J=8.1 Hz, 1 H), 5.91 (s, 1 H), 5.44 (s, 2 H), 3.64 (t, J=8.2 Hz, 2 H), 2.47 (s, 3 H), 1.35 (s, 9 H), 0.95 (t, J=8.6 Hz, 2 H), 0.01 (s, 9 H).
N-text-Butyl-3-(5-methylbenzo[b]thiophen-2-yl)-1H-pyrazole-4-carboxamide
To a solution of N-tert-butyl-3-(5-methylbenzo[b]thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (72 mg, 162 μmol) in dichloromethane (2.5 mL) was added trifluoroacetic acid (370 mg, 250 μL, 3.25 mmol). The mixture was stirred at r.t for 16 h and then concentrated in vacuo. The residue obtained was redissolved in dichloromethane (2.5 mL), methanol (1.3 mL) and ammonium hydroxide (350 μL) and stirred at r.t for 1 h. The mixture was then concentrated in vacuo and purified by chromatography (silica, 40-80% ethyl acetate in hexanes) to give N-tert-butyl-3-(5-methylbenzo[b]thiophen-2-yl)-1H-pyrazole-4-carboxamide (18 mg, 57.4 μmol, 35%) as a white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 8.04 (s, 1 H), 7.76 (d, J=8.2 Hz, 1 H), 7.71 (s, 1 H), 7.63 (s, 1 H), 7.24 (d, J=8.5 Hz, 1 H), 5.92 (s, 1 H), 2.51 (s, 3 H), 1.39 (s, 9 H); MS (EI/CI) m/z: 313.9 [M+H].
N-tert-Butyl-3-(6-methoxypyridin-2-yl)-1H-pyrazole-4-carboxamide
Ethyl 3-(6-methoxypyridin-2-yl)-3-oxopropanoate
To a mixture of methyl 6-methoxypicolinate (200 mg, 1.2 mmol) and ethyl acetate (0.70 mL, 7.18 mmol) in THF (2.4 mL) at −50° C. was quickly added lithium bis(trimethylsilyl)amide (1 M in toluene, 3.6 mL, 3.6 mmol). The mixture was stirred for 30 min, then quenched with acetic acid, and washed with sodium bicarbonate. The organic phase was concentrated in vacuo and purified by chromatography (silica, 10-40% ethyl acetate in hexanes) to give ethyl 3-(6-methoxypyridin-2-yl)-3-oxopropanoate (184 mg, 69%) as a clear oil. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.73 (m, 2 H), 6.99 (d, J=8.2 Hz, 1 H), 4.21 (q, J=7.2 Hz, 2 H), 4.11 (s, 2 H), 3.98 (s, 3 H), 1.24 (t, J=7.1 Hz, 3 H).
(E)-Ethyl 3-ethoxy-2-(6-methoxypicolinoyl)acrylate
Ethyl 3-(6-methoxypyridin-2-yl)-3-oxopropanoate (184 mg, 824 μmol), acetic anhydride (337 mg, 3.3 mmol) and triethyl orthoformate (224 mg, 1.65 mmol) were heated to 110° C. for 6 hours. After which the mixture was cooled and concentrated to give crude (E)-ethyl 3-ethoxy-2-(6-methoxypicolinoyl)acrylate (230 mg) which was used directly in the next step without purification. MS (EI/CI) m/z: 279.8 [M+H].
Ethyl 3-(6-methoxypyridin-2-yl)-1H-pyrazole-4-carboxylate
To a solution of crude (E)-ethyl 3-ethoxy-2-(6-methoxypicolinoyl)acrylate (230 mg, 824 μmol) in ethanol was added hydrazine hydrate (82.5 mg, 1.65 mmol). The mixture was stirred for 72 h, concentrated in vacuo, and then purified by chromatography (silica, 15-50% ethyl acetate in hexanes) to give ethyl 3-(6-methoxypyridin-2-yl)-1H-pyrazole-4-carboxylate (100 mg, 49% over two steps) as a white solid (over two steps). MS (EI/CI) m/z: 248.2 [M+H].
3-(6-Methoxypyridin-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid
To a solution of ethyl 3-(6-methoxypyridin-2-yl)-1H-pyrazole-4-carboxylate (100 mg, 404 μmol) in THF (2 mL) cooled to 0° C. was added sodium hydride (32.4 mg, 809 μmol). The mixture was stirred for 15 min, at which point (2-(Chloromethoxy)ethyl)trimethylsilane (80.9 mg, 485 μmol) was added. After stirring at r.t. for 16 h the mixture was diluted with ethyl acetate and sodium bicarbonate, the phases separated, and the aqueous phase extracted with ethyl acetate. The combined organic layers were washed with water, separated, concentrated in vacuo, and purified by chromatography (silica, 0-50% ethyl acetate in hexanes) to give 3-(6-methoxypyridin-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (54 mg, 155 μmol, 38%) as a white solid. MS (EI/CI) m/z: 349.9 [M+H].
N-tert-Butyl-3-(6-methoxypyridin-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide
3-(6-Methoxypyridin-2-yl)-1 -((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (54 mg, 155 μmol), 2-methylpropan-2-amine (45.2 mg, 618 μmol), N1-((ethylimino)methylene)-N3,N3-dimethylpropane-1,3-diamine hydrochloride (68.1 mg, 355 μmol) and 1H-benzo[d][1,2,3]triazol-1-ol hydrate (54.4 mg, 355 μmol) in DMF (1 mL) were stirred at r.t. for 16 h. The mixture was then quenched with 10% citric acid and diluted with ethyl acetate. The phases were separated and the organic layer washed with sodium bicarbonate and brine, then concentrated in vacuo and purified by chromatography (silica, 5-30% ethyl acetate in hexanes) to give N-tert-butyl-3-(6-methoxypyridin-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (53 mg, 131 μmol, 85%) as a white solid. MS (EI/CI) m/z: 405.1 [M+H].
To a solution of N-tert-butyl-3-(6-methoxypyridin-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (53 mg, 131 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (299 mg, 202 μL, 2.62 mmol). The mixture was stirred at r.t. for 16 h, then concentrated in vacuo. The residue obtained was redissolved in dichloromethane (2 mL), methanol (1 mL) and ammonium hydroxide (300 μL, and stirred at r.t. for 1 h. The mixture was concentrated in vacuo and purified by chromatography (silica, 40-80% ethyl acetate in hexanes) to give N-tert-butyl-3-(6-methoxypyridin-2-yl)-1H-pyrazole-4-carboxamide (27 mg, 98.4 μmol, 75%) as a white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 13.35 (s, 1 H), 9.77 (s, 1 H), 8.19 (s, 1 H), 7.91 (t, J=7.8 Hz, 1 H), 7.63 (d, J=7.7 Hz, 1 H), 6.95 (d, J=8.0 Hz, 1 H), 3.90 (s, 3 H), 1.38 (s, 9 H); MS (EI/CI) m/z: 274.8 [M+H].
(S)-3-(Benzo[d]thiazol-2-yl)-N-(1-hydroxypropan-2-yl)-1H-pyrazole-4-carboxamide
(S)-3-(Benzo[d]thiazol-2-yl)-N-(1-hydroxypropan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide
3-(Benzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (72 mg, 192 μmol), (S)-2-aminopropan-1-ol (57.6 mg, 767 μmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (84.5 mg, 441 μmol) and HOBT (67.5 mg, 441 μmol) in DMF (1 mL) were stirred at r.t. for 16 h. The mixture was quenched with 10% citric acid and diluted with sodium bicarbonate and ethyl acetate. The phases were separated and the organic phase then washed with sodium bicarbonate and brine, concentrated in vacuo. Purification by chromatography (silica, 20-80% ethyl acetate in hexanes) gave (S)-3-(benzo[d]thiazol-2-yl)-N-(1-hydroxypropan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (48 mg, 111 μmol, 58%) as a colorless viscous oil. MS (EI/CI) m/z: 433.0 [M+H].
To a solution of (S)-3-(benzo[d]thiazol-2-yl)-N-(1-hydroxypropan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (48 mg, 111 μmol) in dichloromethane (1.7 mL) was added trifluoroacetic acid (253 mg, 171 μL, 2.22 mmol). The mixture was stirred at r.t. for 16 h, and then concentrated in vacuo. The residue obtained was redissolved in dichloromethane (1.7 mL), methanol (800 μL) and ammonium hydroxide (300 μL) and stirred at r.t. for 1 h. The mixture was then concentrated in vacuo, and the residue obtained triturated with water and filtered. The obtained solid was washed with water and then dried under vacuum to give (S)-3-(benzo[d]thiazol-2-yl)-N-(1-hydroxypropan-2-yl)-1H-pyrazole-4-carboxamide (28 mg, 92.6 μmol, 84%) as a white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 13.87 (s, 1 H), 10.88 (s, 1 H), 8.44 (s, 1 H), 8.18 (d, J=8.1 Hz, 1 H), 8.10 (d, J=8.0 Hz, 1 H), 7.60 (t, J=7.5 Hz, 1 H), 7.53 (t, J=7.7 Hz, 1 H), 4.93 (t, J=6.0 Hz, 1 H), 4.07 (m, 1 H), 3.61 (m, 1 H), 3.52 (m, 1 H), 1.29 (d, J=6.7 Hz, 3 H); MS (EI/CI) m/z: 302.8 [M+H].
Ethyl 3-(4-phenylthiazol-2-yl)-1H-pyrazole-4-carboxylate
Ethyl 3-oxo-3-(4-phenylthiazol-2-yl)propanoate
To a solution of ethyl 4-phenylthiazole-2-carboxylate (900 mg, 3.86 mmol) and ethyl acetate (2.27 mL, 23.1 mmol) in THF (8 mL) cooled to −50° C. was quickly added lithium bis(trimethylsilyl)amide (1 M in toluene, 11.6 mL, 11.6 mmol). The mixture was stirred for 30 min, quenched with acetic acid, then diluted with water and sodium bicarbonate. The mixture was extracted with ethyl acetate and the combined organic extracts were concentrated in vacuo and purified by chromatography (silica, 10-50% ethyl acetate in hexanes) to give ethyl 3-oxo-3-(4-phenylthiazol-2-yl)propanoate (890 mmol, 3.23 mmol, 84%). MS (EI/CI) m/z: 276.1 [M+H].
(Z)-Ethyl 3-(dimethylamino)-2-(4-phenylthiazole-2-carbonyl)acrylate
To a solution of ethyl 3-oxo-3-(4-phenylthiazol-2-yl)propanoate (890 mmol, 3.23 mmol) in ethanol (9 mL) was added dimethylformamide dimethyl acetal (385 mg, 3.23 mmol). The mixture was heated to 80° C. for 15 h and then concentrated in vacuo. Purification by chromatography (silica, 20-100% ethyl acetate in hexanes) gave (Z)-ethyl 3-(dimethylamino)-2-(4-phenylthiazole-2-carbonyl)acrylate (720 mg, 67%). 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.95 (d, J=1.6 Hz, 2 H), 7.93 (s, 1 H), 7.45 (t, J=7.3 Hz, 2 H), 7.37 (t, J=7.4 Hz, 1 H), 4.13 (q, J=7.0 Hz, 2 H), 3.30 (br. s, 3 H), 3.01 (br. s, 3 H), 1.03 (t, J=7.1 Hz, 3 H).
To a solution of (Z)-ethyl 3-(dimethylamino)-2-(4-phenylthiazole-2-carbonyl)acrylate (700 mg, 2.12 mmol) in ethanol (11 mL) was added hydrazine hydrate (117 mg, 2.33 mmol). Additional ethanol (5 mL) was added to the thick suspension after ˜1 min and the mixture was stirred for a further 15 min. The mixture was diluted EtOH (5 mL), and then cooled to 0° C. The solid precipitate was collected by filtration and washed with cold EtOH, then diethyl ether, and finally dried to give a white solid. An additional amount of product was obtained by first concentrating the organic filtrate in vacuo, and then purifying the residue by chromatography (silica, 15-60% ethyl acetate in hexanes). This material was combined with the previously obtained solid to give ethyl 3-(4-phenylthiazol-2-yl)-1H-pyrazole-4-carboxylate (575 mg, 86%) as a white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 8.11 (s, 1 H), 7.98 (d, J=7.9 Hz, 2 H), 7.73 (s, 1 H), 7.49 (t, J=7.6 Hz, 2 H), 7.41 (t, J=7.5 Hz, 1 H), 4.45 (q, J=7.3 Hz, 2 H), 1.46 (t, J=7.2 Hz, 3 H); MS (EI/CI) m/z: 299.8 [M+H].
Ethyl 3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-pyrazole-4-carboxylate
Ethyl 4,5,6,7-tetrahydrobenzo[d]thiazole-2-carboxylate
To a solution of 2-chlorocyclohexanone (2 g, 15.1 mmol) in ethanol (18 mL) was added ethyl 2-amino-2-thioxoacetate (2.11 g, 15.8 mmol). The mixture was sealed in two identical microwave vials and each one was irradiated at 150° C. for 1.5 h. Upon cooling the mixture was concentrated in vacuo and directly purified by chromatography (silica, 5-45% ethyl acetate in hexanes) to give ethyl 4,5,6,7-tetrahydrobenzo[d]thiazole-2-carboxylate (826 mg, 26%) as a yellow oil. MS (EI/CI) m/z: 212.3 [M+H].
Ethyl 3-oxo-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)propanoate
To a solution of ethyl 4,5,6,7-tetrahydrobenzo[d]thiazole-2-carboxylate (240 mg, 1.14 mmol) in THF (2.3 mL) at −50° C. was added ethyl acetate (600 mg, 667 μL, 6.82 mmol) followed by lithium bis(trimethylsilyl)amide (1M in THF, 3.41 mL, 3.41 mmol). The mixture was stirred for 30 min and then quenched with acetic acid, diluted with water and saturated sodium bicarbonate solution and extracted with ethyl acetate. The combined extracts were concentrated in vacuo and the residue obtained was purified by chromatography (silica, 0-20% ethyl acetate in hexanes) to give ethyl 3-oxo-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)propanoate (110 mg, 434 μmol, 38%) as a yellow liquid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 4.24 (q, J=7.1 Hz, 2 H), 4.13 (s, 1 H), 2.89 (m, 4 H), 1.92 (m, 4 H), 1.29 (t, J=7.3 Hz, 3 H).
(Z)-Ethyl 3-(dimethylamino)-2-(4,5,6,7-tetrahydrobenzo[d]thiazole-2-carbonyl)acrylate
To a solution of ethyl 3-oxo-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)propanoate (680 mg, 2.68 mmol) in ethanol (7.7 mL) was added dimethylformamide dimethyl acetal (352 mg, 2.95 mmol). The mixture was heated to 80° C. for 3 h, cooled to r.t., concentrated in vacuo, and then purified by chromatography (silica, 20-100% ethyl acetate in hexanes) to give (Z)-ethyl 3-(dimethylamino)-2-(4,5,6,7-tetrahydrobenzo[d]thiazole-2-carbonyl)acrylate (464 mg, 56%). 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.82 (s, 1 H), 4.14 (m, 2 H), 3.19 (br. s, 3 H), 2.98 (br. s, 3 H), 2.85 (m, 4 H), 1.89 (m, 4 H), 1.12 (t, J=7.1 Hz, 3 H).
To a solution of (Z)-ethyl 3-(dimethylamino)-2-(4,5,6,7-tetrahydrobenzo[d]thiazole-2-carbonyl)acrylate (464 mg, 1.5 mmol) in ethanol (15 ml) was added a hydrazine monohydrate (82.9 mg, 1.66 mmol) in ethanol (1 mL) and the mixture stirred for 1 h. The thick suspension was diluted with cold diethyl ether (10 mL), filtered, and the solid obtained was washed with an additional portion of cold diethyl ether. The material was then dried in vacuo to give ethyl 3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-pyrazole-4-carboxylate (370 mg, 88%). 1H NMR (400 MHz, DMSO-d) δ ppm 8.15 (s, 1 H), 4.26 (q, J=7.0 Hz, 2 H), 2.80 (m, 4 H), 1.84 (m, 4 H), 1.23 (t, J=7.0 Hz, 3 H); MS (EI/CI) m/z: 277.8 [M+H].
Ethyl 3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1H-pyrazole-4-carboxylate
Ethyl 5,6-dihydro-4H-cyclopenta[d]thiazole-2-carboxylate
Ethyl 2-amino-2-thioxoacetate (1.4 g, 10.05 mmol) and 2-chlorocyclopentanone (1.18 g, 9.99 mmol) in ethanol (10 mL) were placed in a microwave vial and irradiated at 150° C. for 1.5 h. The mixture was cooled, concentrated in vacuo and then purified by chromatography (silica, 10-50% ethyl acetate in hexanes) to give ethyl 5,6-dihydro-4H-cyclopenta[d]thiazole-2-carboxylate (850 mg, 43%). 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 4.48 (q, J=7.1 Hz, 2 H), 3.02 (t, J=7.3 Hz, 2 H), 2.95 (t, J=7.5 Hz, 2 H), 2.57 (m, 2 H), 1.46 (t, J=7.1 Hz, 3 H).
Ethyl 3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-3-oxopropanoate
To a solution of ethyl 5,6-dihydro-4H-cyclopenta[d]thiazole-2-carboxylate (850 mg, 4.31 mmol) and ethyl acetate (2.53 mL, 25.9 mmol) in THF (8.6 mL) at −50° C. was quickly added lithium bis(trimethylsilyl)amide (1M in toluene, 12.9 mL, 12.9 mmol). The mixture was stirred for 30 min before being quenched with acetic acid. After warming to room temperature it was diluted with water and saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, and concentrated in vacuo. The residue obtained was purified by chromatography (silica, 10-50% ethyl acetate in hexanes) to give ethyl 3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-3-oxopropanoate (900 mg, 87%) as an orange oil. MS (EI/CI) m/z: 239.7 [M+H].
(Z)-Ethyl 2(5,6-dihydro-4H-cyclopenta[d]thiazole-2-carbonyl)-3-(dimethylamino)arcylate
To a solution of ethyl 3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-3-oxopropanoate (750 mg, 3.13 mmol) in ethanol (9 mL) was added dimethylformamide dimethyl acetal (411 mg, 3.45 mmol). The mixture was heated to 80° C. for 3 h then cooled, concentrated in vacuo, and purified by chromatography (silica, 33-100% ethyl acetate in hexanes) to give (Z)-ethyl 2-(5,6-dihydro-4H-cyclopenta[d]thiazole-2-carbonyl)-3-(dimethylamino)acrylate (472 mg, 51%) as an oil. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.84 (s, 1 H), 4.15 (q, J=7.1 Hz, 2 H), 3.20 (br, s, 6 H), 2.99 (t, J=7.1 Hz, 2 H), 2.89 (t, J=7.4 Hz, 2 H), 2.53 (m, 2 H), 1.13 (t, J=7.4 Hz, 3 H).
To a solution of (Z)-ethyl 2-(5,6-dihydro-4H-cyclopenta[d]thiazole-2-carbonyl)-3-(dimethylamino)acrylate (472 mg, 1.6 mmol) in ethanol (16 mL) was added hydrazine hydrate (88 mg, 1.76 mmol) in ethanol (1 mL). The thick suspension was stirred at r.t. for 1 h, diluted with diethyl ether, and collected by filtration, washed with ether, and then then dried to give ethyl 3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1H-pyrazole-4-carboxylate (362 mg, 86%) as an off-white solid. 1H NMR (400 MHz, DMSO-d) δ ppm 8.15 (s, 1 H), 4.26 (q, J=7.4 Hz, 2 H), 2.97 (t, J=7.1 Hz, 2 H), 2.64 (t, J=7.7 Hz, 2 H), 2.52 (m, 2 H), 1.28 (t, J=7.1 Hz, 3 H); MS (EI/CI) m/z: 263.8 [M+H].
N-tert-Butyl-3-(4-phenylthiazol-2-yl)-1H-pyrazole-4-carboxamide
Ethyl 3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate
To a solution of ethyl 3-(4-phenylthiazol-2-yl)-1H-pyrazole-4-carboxylate (560 mg, 1.87 mmol) in THF (9.4 mL) cooled to 0° C., was added sodium hydride (150 mg, 3.74 mmol). The mixture was stirred for 15 min and then (2-(Chloromethoxy)ethyl)trimethylsilane (374 mg, 2.24 mmol) was added. After stirring at r.t. for 16 h the reaction mixture was quenched with saturated sodium bicarbonate solution and then extracted into ethyl acetate. The combined organic extracts were concentrated in vacuo and purified by chromatography (silica, 5-25% ethyl acetate in hexanes) to give ethyl 3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate (662 mg, 1.54 mmol, 82%) as a colorless oil. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 8.19 (s, 1 H), 7.99 (d, J=8.3 Hz, 2 H), 7.61 (s, 1 H), 7.42 (t, J=7.9 Hz, 2 H), 7.33 (t, J=7.4 Hz, 1 H), 5.52 (s, 2 H), 4.34 (q, J=7.18 Hz, 2 H), 3.67 (t, J=8.4 Hz, 2 H), 1.32 (t, J=7.3 Hz, 3 H), 0.96 (t, J=8.7, 2 H), 0.01 (s, 9 H).
3-(4-Phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid
To a solution of ethyl 3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate (790 mg, 1.84 mmol) in dioxane (26 mL) was added 1N sodium hydroxide (9.2 mL, 9.2 mmol). The mixture was stirred at r.t. for 16 h and then quenched with 10% HCl (˜pH 6). The precipitate obtained by filtration, was washed with water and ether, and then dried to give 3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (349 mg, 869 μmol, 47%) as a white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 8.38 (s, 1 H), 7.84 (d, J=8.0 Hz, 2 H), 7.56 (s, 1 H), 7.48 (t, J=6.8 Hz, 2 H), 7.41 (t, J=7.3 Hz, 1 H), 5.49 (s, 2 H), 3.66 (t, J=8.4 Hz, 2 H), 0.96 (t, J=8.4, 2 H), 0.01 (s, 9 H).
Step 3 N-tert-Butyl-3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide
3-(4-Phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (115 mg, 286 μmol), 2-methylpropan-2-amine (83.8 mg, 1.15 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (126 mg, 659 μmol) and HOBT (101 mg, 659 μmol) in DMF (1.2 mL) were stirred at r.t. for 16 h. After which the reaction mixture was quenched with 10% citric acid and then diluted with ethyl acetate and sodium bicarbonate. The phases were separated and the organic phase washed with additional sodium bicarbonate and brine, dried, filtered and concentrated in vacuo. Purification by chromatography (silica, 10-30% ethyl acetate in hexanes) gave N-tert-butyl-3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (115 mg, 252 μmol, 88%) as a white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 9.92 (s, 1 H), 8.31 (s, 1 H), 7.80 (d, J=7.7 Hz, 2 H), 7.42 (m, 3 H), 5.45 (s, 2 H), 3.53 (t, J=8.4 Hz, 2 H), 1.38 (s, 9 H), 0.95 (t, J=8.2 Hz, 2 H), 0.01 (s, 9 H).
To a solution of N-tert-butyl-3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (115 mg, 252 μmol) in dichloromethane (3.8 mL) was added trifluoroacetic acid (574 mg, 388 μL, 5.04 mmol). The mixture was stirred at r.t. for 16 h, then concentrated in vacuo. The residue obtained was redissolved in dichloromethane (3.8 mL), methanol (1.9 mL) and ammonium hydroxide (600 μL) and then stirred at r.t. for 1 h. This mixture was then concentrated in vacuo, triturated with water and filtered. The solid obtained was washed with water and ether. Further purification by chromatography (silica, 20-60% ethyl acetate in hexanes) gave N-tert-butyl-3-(4-phenylthiazol-2-yl)-1H-pyrazole-4-carboxamide (56 mg, 172 μmol, 68%) as an off-white solid. 1H NMR (400 MHz, DMSO-d6) δ ppm 13.71 (s, 1 H), 9.55 (s, 1 H), 8.34 (s, 1 H), 8.10 (s, 1 H), 7.91 (d, J=7.6 Hz, 2 H), 7.50 (t, J=7.6 Hz, 2 H), 7.42 (t, J=7.4 Hz, 1 H), 1.33 (s, 9 H); MS (EI/CI) m/z: 326.9 [M+H].
N-Isopropyl-3-(4-phenylthiazol-2-yl)-1H-pyrazole-4-carboxamide
N-Isopropyl-3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide
To a solution of 3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (89 mg, 222 μmol) in DMF (1 mL) was added HATU (101 mg, 266 μmol), DIPEA (85.9 mg, 116 μL, 665 μmol) and propan-2-amine (39.3 mg, 665 μmol). The mixture was stirred at r.t. for 16 h, then quenched with 10% citric acid and diluted with sodium bicarbonate and ethyl acetate. The organic layer was collected, concentrated in vacuo, and then purified by chromatography (silica, 15-45% ethyl acetate in hexanes) to give N-isopropyl-3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (92 mg, 208 μmol, 94%) as a white solid. MS (EI/CI) m/z: 443.2 [M+H].
To a solution of N-isopropyl-3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (92 mg, 208 μmol) in dichloromethane was added trifluoroacetic acid (474 mg, 320 μl, 4.16 mmol). The mixture was stirred at r.t. for 16 h and then concentrated in vacuo. The residue obtained was redissolved in dichloromethane (3.2 mL), methanol (1.8 mL) and ammonium hydroxide (500 μL) and stirred at r.t. for 1 h. The mixture was concentrated in vacuo and directly purified by chromatography (silica, 50-90% ethyl acetate in hexanes) to give N-isopropyl-3-(4-phenylthiazol-2-yl)-1H-pyrazole-4-carboxamide (40 mg, 128 μmol, 62%) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ ppm 13.86 (s, 1 H), 10.27 (d, J=8.3 Hz, 1 H), 8.50 (s, 1 H), 8.27 (s, 1 H), 8.05 (d, J=7.5 Hz, 2 H), 7.63 (t, J=7.5 Hz, 2 H), 7.54 (t, J=7.5 Hz, 1 H), 4.28 (m, 1 H), 1.28 (d, J=6.7 Hz, 6 H); MS (EI/CI) m/z: 312.8 [M+H].
N-tert-Butyl-3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1H-pyrazole-4-carboxamide
Ethyl 3-5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1-((2-trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate
To a solution of ethyl 3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1H-pyrazole-4-carboxylate (342 mg, 1.3 mmol) in THF (6.5 mL) cooled to 0° C. was added sodium hydride (104 mg, 2.6 mmol). After stirring for 15 min (2-(chloromethoxy)ethyl)trimethylsilane (260 mg, 1.56 mmol) was added, and the mixture was warmed to r.t. After 16 h, the mixture was quenched with sodium bicarbonate and diluted with ethyl acetate. The organic phase was separated and washed with sodium bicarbonate and brine, concentrated in vacuo, and purified by chromatography (silica, 10-30% ethyl acetate in hexanes) to give ethyl 3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate (380 mg, 966 μmol, 74%) as an orange oil. MS (EI/CI) m/z: 394.0 [M+H].
3-(5,6-Dihydro-4H-cyclopenta[d]trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid
To a solution of ethyl 3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate (437 mg, 1.11 mmol) in dioxane (15.9 mL) was added 1M sodium hydroxide (5.55 mL, 5.55 mmol). The mixture was stirred at r.t. for 16 h, then acidified with 10% HCl (to ˜pH 6). The mixture was diluted with water and ethyl acetate, and the phases separated. The organic phase was washed with water and brine, then concentrated in vacuo. Purification by chromatography (silica, 20-60% ethyl acetate in hexanes) gave 3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (306 mg, 837 μmol, 75%) as a white solid. MS (EI/CI) m/z: 365.9 [M+H].
N-tert-Butyl-3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide
3-(5,6-Dihydro-4H-cyclopenta[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (100 mg, 274 μmol), 2-methylpropan-2-amine (80.0 mg, 1.09 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (121 mg, 629 μmol) and HOBT (96.4 mg, 629 μmol) in DMF (1.5 mL) were stirred at r.t. for 16 h. The reaction mixture was quenched with 10% citric acid and then diluted with sodium bicarbonate and ethyl acetate. The organic phase was separated and then washed with sodium bicarbonate and water, concentrated in vacuo, and purified by chromatography (silica, 10-30% ethyl acetate in hexanes) to give N-tert-butyl-3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (105 mg, 250 μmol, 91%) as a white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 10.94 (s, 1 H), 8.32 (s, 1 H), 5.43 (s, 1 H), 3.61 (t, J=8.4 Hz, 2 H), 2.98 (t, J=7.2 Hz, 2 H), 2.89 (t, J=7.2 Hz, 2 H), 2.58 (m, 2 H), 1.51 (s, 9 H), 0.95 (t, J=8.4 Hz, 2 H), 0.01 (s, 9 H).
To a solution of N-tert-butyl-3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (105 mg, 250 μmol) in dichloromethane (3.9 mL) was added trifluoroacetic acid (569 mg, 385 μL, 4.99 mmol). The mixture was stirred at r.t for 16 h, then concentrated in vacuo. The residue was redissolved in dichloromethane (3.5 mL), methanol (1.7 mL) and ammonium hydroxide (600 μL) and stirred at r.t. for 1 h. The mixture was then concentrated in vacuo, the residue obtained was triturated with water and collected by filtration. The solid was washed with water and ether and then dried under vacuum to give N-tert-butyl-3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1H-pyrazole-4-carboxamide (62 mg, 214 μmol, 86%) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ ppm 13.53 (s, 1 H), 10.70 (s, 1 H), 8.25 (s, 1 H), 2.94 (t, J=7.0 Hz, 2 H), 2.83 (t, J=7.2 Hz, 2 H), 2.50 (m, 2 H), 1.41 (s, 9 H); MS (EI/CI) m/z: 290.9 [M+H].
N-tert-Butyl-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-pyrazole-4-carboxamide
Ethyl 3-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1-((2-trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate
To a solution of ethyl 3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-pyrazole-4-carboxylate (359 mg, 1.29 mmol) in (2-(chloromethoxy)ethyl)trimethylsilane (259 mg, 1.55 mmol) cooled to 0° C. was added sodium hydride (104 mg, 2.59 mmol). The mixture was stirred for 15 min and (2-(Chloromethoxy)ethyl)trimethylsilane (259 mg, 1.55 mmol) was then added. After stirring at r.t. for 16 h, the reaction mixture was quenched with aqueous sodium bicarbonate and diluted with ethyl acetate. The organic phase was separated, washed with sodium bicarbonate and brine, concentrated in vacuo and purified by chromatography (silica, 10-30% ethyl acetate and hexanes) to give ethyl 3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate (300 mg, 736 μmol, 57%) as a colorless oil. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 8.07 (s, 1 H), 5.84 (s, 1 H), 4.35 (q, J=7.1 Hz, 2 H), 3.58 (t. J=8.4 Hz, 2 H), 2.94 (m, 4 H), 1.98 (m, 4 H), 1.38 (t, J=7.1 Hz, 3 H), 0.90 (t, J=8.4 Hz, 2 H), 0.01 (s, 9 H).
3-4,5,6,7-Tetrahydrobenzo[d]thiazol-2-yl)-1-((2-trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid
To a solution of ethyl 3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylate (350 mg, 859 μmol) in dioxane (12.3 mL) was added 1N sodium hydroxide (4.29 mL, 4.29 mmol). The mixture was stirred at r.t. for 16 h then acidified with 10% HCl (to ˜pH 6) and then diluted with water and ethyl acetate. The organic phase was separated washed with water and brine, and then concentrated in vacuo. Purification by chromatography (silica 20-60% ethyl acetate in hexanes) gave 3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (273 mg, 719 μmol, 84%) as a white solid. MS (EI/CI) m/z: 379.9 [M+H].
N-tert-Butyl-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1-((2-trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide
3-(4,5,6,7-Tetrahydrobenzo[d]thiazol-2-yl)-1((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (100 mg, 263 μmol), 2-methylpropan-2-amine (77.1 mg, 1.05 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (116 mg, 606 μmol) and HOBT (92.8 mg, 606 μmol) in DMF (1.5 mL) were stirred at r.t. for 16 h. The reaction mixture was quenched with 10% citric acid and then diluted with sodium bicarbonate and ethyl acetate. The organic phase was separated, washed with sodium bicarbonate and water, then concentrated in vacuo. Purification by chromatography (silica, 10-30% ethyl acetate in hexanes) gave N-tert-butyl-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (102 mg, 235 μmol, 89%) as a white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 10.89 (s, 1 H), 8.28 (s, 1 H), 5.42 (s, 2 H), 3.61 (t, J=8.6 Hz, 2 H), 2.83 (m, 4 H), 1.92 (m, 2 H). 1.52 (s, 9 H), 0.95 (t, J=8.4 Hz, 2 H), 0.01 (s, 9 H).
To a solution of N-tert-butyl-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (102 mg, 235 μmol) in dichloromethane (3.6 mL) was added trifluoroacetic acid (535 mg, 362 μL, 4.69 mmol). The mixture was stirred at r.t. for 16 h then concentrated in vacuo. The residue obtained was redissolved in dichloromethane (3.5 mL), methanol (1.7 mL) and ammonium hydroxide (550 μL). This mixture was stirred at r.t. for 1 h then concentrated in vacuo, and the residue obtained triturated with water. The solid was collected by filtration and washed with water and ether, then dried under vacuum to give N-butyl-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-pyrazole-4-carboxamide (43 mg, 141 μmol, 60%) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ ppm 13.54 (s, 1 H), 10.68 (s, 1 H), 8.26 (s, 1 H), 2.80 (m, 4 H), 1.84 (m, 4 H), 1.43 (s, 9 H); MS (EI/CI) m/z: 304.9 [M+H].
N-Cyclopentyl-3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide
To a solution of 3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (58 mg, 144 μmol) in DMF (1 mL) was added cyclopentanamine (36.9 mg, 433 μmol), HATU (65.9 mg, 173 μmol) and DIPEA (56.0 mg, 75.7 μL, 433 μmol). The mixture was stirred at r.t. for 16 h, then quenched with 10% citric acid and then diluted with ethyl acetate. The organic phase was separated, washed with sodium bicarbonate and brine, then concentrated in vacuo. Purification by chromatography (silica, 15-35% ethyl acetate in hexanes) to give N-cyclopentyl-3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (58 mg, 124 μmol, 86%) as a white solid. MS (EI/CI) m/z: 469.1 [M+H].
To a solution of N-cyclopentyl-3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (58 mg, 124 μmol) in dichloromethane (1.9 mL) was added trifluoroacetic acid (282 mg, 191 μL, 2.47 mmol). The mixture was stirred at r.t for 16 h then concentrated in vacuo. The residue obtained was redissolved in dichloromethane (2 mL), methanol (1 mL) and ammonium hydroxide (300 μL) and stirred at r.t. for 1 h. The mixture was then concentrated in vacuo, and the residue obtained was triturated with water and collected by filtration. The solid was washed with water and ether then dried to give N-cyclopentyl-3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (58 mg, 124 μmol, 100%) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ ppm 13.77 (s, 1 H), 10.22 (d, J=7.5 Hz, 1 H), 8.41 (s, 1 H), 8.18 (s, 1 H), 7.93 (d, J=7.5 Hz, 2 H), 7.54 (t, J=7.7 Hz, 2 H), 7.46 (t, J=7.3 Hz, 1 H), 4.31 (m, 1 H), 1.95 (m, 2 H), 1.61 (m, 2 H), 1.49 (m, 4 H); MS (EI/CI) m/z: 339.9 [M+H].
3-(4-Phenylthiazol-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazole-4-carboxamide
3-(4-Phenylthiazol-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide
To a solution of 3-(4-phenylthiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (58 mg, 144 μmol) in DMF (1 mL) was added tetrahydro-2H-pyran-4-amine (43.8 mg, 433 μmol), HATU (65.9 mg, 173 μmol) and DIPEA (56.0 mg, 75.7 μL, 433 μmol). The mixture was stirred at r.t. for 16 h, then quenched with 10% citric acid and diluted with ethyl acetate. The organic phase was separated, washed with sodium bicarbonate and brine, then concentrated in vacuo. Purification by chromatography (silica, 15-100% ethyl acetate in hexanes) gave 3-(4-phenylthiazol-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (67 mg, 138 μmol, 96%) as a white solid. MS (EI/CI) m/z: 485.1 [M+H].
To a solution of 3-(4-phenylthiazol-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (67 mg, 138 μmol) in dichloromethane (2.1 mL) was added trifluoroacetic acid (315 mg, 213 μL, 2.76 mmol). The mixture was stirred at r.t. for 16 h, then concentrated in vacuo. The residue obtained was redissolved in dichloromethane (2 mL), methanol (1 mL) and ammonium hydroxide (300 μL) and stirred at r.t for 1 h. The mixture was then concentrated in vacuo, and the solid obtained triturated with water, filtered, washed with water and ether, then dried to give 3-(4-phenylthiazol-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazole-4-carboxamide (33.7 mg, 95.1 μmol, 69%) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ ppm 13.78 (s, 1 H), 10.24 (d, J=7.7 Hz, 1 H), 8.42 (s, 1 H), 8.18 (s, 1 H), 7.95 (d, J=7.7 Hz, 2 H), 7.52 (t, J=7.8 Hz, 2 H), 7.45 (t, J=7.3 Hz, 1 H), 4.10 (m, 1 H), 3.83 (d, J=12.1 Hz), 3.37 (q, J=11.8 Hz, 2 H), 1.83 (d, J=12.2 Hz, 2 H), 1.52 (q, J=11.6 Hz, 2 H); MS (EI/CI) m/z: 355.0 [M+H].
3-(5,6-Dihydro-4H-cyclopenta[d]thiazol-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazole-4-carboxamide
3-(5,6-Dihydro-4H-cyclopenta[d]thiazol-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide
To a solution of 3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (100 mg, 274 μmol) in DMF (2 mL) was added tetrahydro-2H-pyran-4-amine (83.0 mg, 821 μmol, HATU (156 mg, 410 μmol) and DIPEA (106 mg, 143 μL, 821 μmol). The mixture was stirred at r.t. for 16 h, quenched with 10% citric acid, and then diluted with ethyl acetate and sodium bicarbonate. The organic phase was separated, washed with sodium bicarbonate and brine, concentrated in vacuo and then purified by chromatography (silica, 40-80% ethyl acetate in hexanes) to give 3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (121 mg, 270 μmol, 99%) as an off-white solid. MS (EI/CI) m/z: 449.1 [M+H].
To a solution of 3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (121 mg, 270 μmol) in dichloromethane (4.2 mL) was added trifluoroacetic acid (615 mg, 416 μL, 5.39 mmol). The mixture was stirred at r.t. for 16 h then concentrated in vacuo. The residue obtained was redissolved in dichloromethane (4 mL), methanol (2 mL) and ammonium hydroxide (600 μL) and stirred at r.t for 1 h. The mixture was then concentrated in vacuo, triturated with water, filtered and washed with water and ether to give 3-(5,6-dihydro-4H-cyclopenta[d]thiazol-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazole-4-carboxamide (68 mg, 203 μmol, 75%) as a white solid. 1H NMR (400 MHz, DMSO-d) δ ppm 13.57 (s, 1 H), 10.90 (s, 1H), 8.37 (s, 1 H), 4.07 (m, 1 H), 3.89 (m, 2 H), 3.54 (t, J=10.2 Hz, 2 H), 2.99 (t, J=7.4 Hz, 2 H), 2.89 (t, J=6.7 Hz, 2 H), 1.95 (d, J=13.2 Hz, 2 H), 1.54 (m, 2 H); MS (EI/CI) m/z: 319.0 [M+H].
N-(Tetrahydro-2H-pyran-4-yl)-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-pyrazole-4-carboxamide
N-(Tetrahydro-2H-pyran-4-yl)-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide
To a solution of 3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (82 mg, 216 μmol) in DMF (2 mL) was added tetrahydro-2H-pyran-4-amine (65.6 mg, 648 μmol), HATU (123 mg, 324 μmol) and DIPEA (83.8 mg, 113 μL, 648 μmol). The mixture was stirred at r.t. for 16 h then quenched with 10% citric acid and diluted with ethyl acetate and sodium bicarbonate. The organic phase was separated and washed with sodium bicarbonate and brine, then concentrated in vacuo and purified by chromatography (silica, 50-90% ethyl acetate in hexanes) to give N-(tetrahydro-2H-pyran-4-yl)-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (79 mg, 171 μmol, 79%) as a white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 11.25 (s, 1 H), 8.24 (s, 1 H), 5.44 (s, 2 H), 4.21 (m, 1 H), 4.03 (m, 2 H), 3.62 (t, J=8.5 Hz, 2 H), 3.58 (m, 2 H), 2.84 (m, 4 H), 2.06 (d, J=13.4 Hz, 2 H), 1.93 (m, 4 H), 1.71 (m, 2 H), 0.95 (t, J=8.3 Hz, 2 H), 0.01 (s, 9 H).
To a solution of N-(tetrahydro-2H-pyran-4-yl)-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (79 mg, 171 μmol) in dichloromethane (2.6 mL) was added trifluoroacetic acid (389 mg, 263 μL, 3.41 mmol). The mixture was stirred at r.t. for 16 h then concentrated in vacuo. The residue was redissolved with dichloromethane (3 mL), methanol (1.5 mL) and ammonium hydroxide (500 μL) and stirred at r.t for 1 h. The mixture was then concentrated in vacuo, triturated with water, filtered and washed with water and ether to give N-(tetrahydro-2H-pyran-4-yl)-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-pyrazole-4-carboxamide (40 mg, 120 μmol, 71%) as a white solid. 1H NMR (400 MHz, DMSO-d) δ ppm 13.43 (s, 1 H), 10.93 (s, 1 H), 8.33 (s, 1 H), 4.00 (m, 1 H), 3.89 (m, 2 H), 3.46 (t, J=11.7 Hz, 2 H), 2.80 (m 4 H), 1.88 (m, 6 H), 1.52 (m, 2 H); MS (EI/CI) m/z: 333.0 [M+H].
N-(1-Methylpiperidin-4-yl)-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-pyrazole-4-carboxamide
N-(1-Methylpiperidin-4-yl)-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide
To a solution of 3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxylic acid (82 mg, 216 μmol) in DMF (2.00 mL) was added 1-methylpiperidin-4-amine (74.0 mg, 648 μmol), HATU (123 mg, 324 μmol) and DIPEA (83.8 mg, 113 μl, 648 μmol). The mixture was stirred at r.t. for 16 h, quenched with 10% citric acid, and diluted with ethyl acetate and sodium bicarbonate. The organic phase was separated washed with sodium bicarbonate and brine, concentrated in vacuo, and purified by chromatography (silica, 5-10% methanol in dichloromethane to give N-(1-methylpiperidin-4-yl)-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (55 mg, 116 μmol, 54%) as a white solid. MS (EI/CI) m/z: 476.2 [M+H].
To a solution of N-(1-methylpiperidin-4-yl)-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carboxamide (55 mg, 116 μmol) in dichloromethane (1.8 mL) was added trifluoroacetic acid (264 mg, 178 μL, 2.31 mmol). The mixture was stirred at r.t. for 16 h then concentrated in vacuo. The residue obtained was redissolved in dichloromethane (2 mL), methanol (1 mL) and ammonium hydroxide (300 μL), and stirred at r.t. for 1 h. The mixture was then concentrated in vacuo, and the residue triturated with ether. NMR analysis showed the presence of the TFA salt. A few drops of 1N NaOH were added until ˜pH 8, then the mixture was diluted with dichloromethane and washed with water. The organic layer was collected, dried over MgSO4, filtered and dried under vacuum to give N-(1-methylpiperidin-4-yl)-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-pyrazole-4-carboxamide (16 mg, 46.3 μmol, 40%) as an off-white solid. 1H NMR (400 MHz, METHANOL-d) δ ppm 8.26 (s, 1 H), 3.90 (m, 1 H), 2.97 (d, J=11.6 Hz, 2 H), 2.88 (m, 1 H), 2.37 (s, 3 H), 2.30 (t, J=10.7 Hz, 2 H), 2.12 (d, J=13.1 Hz, 2 H), 1.96 (m, 4 H), 1.75 (q, J=11.6 Hz, 2 H); MS (EI/CI) m/z: 346.0 [M+H].
Tetrahydro-pyran-4-carboxylic acid (3-benzothiazol-2-yl-1H-pyrazol-4-yl)-amide
Tetrahydro-pyran-4-carboxylic acid (3-benzothiazol-2-yl-1H-pyrazol-4-yl)-amide (92 mg, 30%) was synthesized as white solid from 3-benzothiazol-2-yl-1H-pyrazol-4-ylamine (200 mg, 0.926 mmol) and tetrahydro-pyran-4-carbonyl chloride (0.35 mL, 3.06 mmol) following the procedure described for N-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide (Example 29). MS (EI/CI) m/z: 329.4 [M+H].
Cyclobutanecarboxylic acid (3-benzothiazol-2-yl-1H-pyrazol-4-yl)-amide
Cyclobutanecarboxylic acid (3-benzothiazol-2-yl-1H-pyrazol-4-yl)-amide (108 mg, 39%) was synthesized as white solid from 3-benzothiazol-2-yl-1H-pyrazol-4-ylamine (200 mg, 0.926 mmol) and cyclobutanecarbonyl chloride (0.35 mL, 3.06 mmol) following the procedure described for N-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide (Example 29). MS (EI/CI) m/z: 299.4 [M+H].
Acetic acid 2-(3-benzothiazol-2-yl-1H-pyrazol-4-ylcarbamoyl)-2-methyl-propyl ester
Acetic acid 2-(3-benzothiazol-2-yl-1H-pyrazol-4-ylcarbamoyl)-2-methyl-propyl ester (210 mg, 51%) was synthesized as off white solid from 3-benzothiazol-2-yl-1H-pyrazol-4-ylamine (250 mg, 1.157 mmol) and acetic acid 2-chlorocarbonyl-2-methyl-propyl ester (Prepared as described in Example 33) (680 mg, 3.82 mmol) following the procedure described for N-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide (Example 29). MS (EI/CI) m/z: 359.2 [M+H].
N-(3-Benzothiazol-2-yl-1H-pyrazol-4-yl)-3-hydroxy-2,2-dimethyl-propionamide
N-(3-Benzothiazol-2-yl-1H-pyrazol-4-yl)-3-hydroxy-2,2-dimethyl-propionamide (65.0 mg, 74%) was synthesized as white solid from acetic acid 2-(3-benzothiazol-2-yl-1H-pyrazol-4-ylcarbamoyl)-2-methyl-propyl ester (100 mg, 0.279 mmol) following the procedure described for N-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-3-hydroxy-2,2-dimethyl-propionamide (Example 34). MS (EI/CI) m/z: 317.2 [M+H].
4-(5-Methyl-1H-benzoimidazol-2-yl)-thiazole-5-carboxylic acid tert-butylamide
2-(5-Bromo-thiazol-4-yl)-5-methyl-1H-benzoimidazole
A solution of 5-bromo-thiazole-4-carboxylic acid (1.37 g, 6.59 mmol, commercially available from Combi-blocks), diisopropylethylamine (1.49 mL, 8.57 mmol), 4-methyl-benzene-1,2-diamine (0.88 g, 7.25 mmol) in DMF (11 mL) at room temperature was treated with TBTU (2.75 g, 8.57 mmol) and the mixture stirred at room temperature for 18 h. The solvents were then evaporated under vacuum and the residue partitioned between ethyl acetate and saturated solution of sodium bicarbonate, the organics were collected and dried with magnesium sulfate and then concentrated under vacuum. The crude mixture was dissolved in acetic acid (12 mL), transferred to microwave vial and the mixture was heated at 150° C. in a microwave reactor for 1.5 h. After solvent evaporation, the residue was dissolved in dichloromethane, washed with saturated solution of sodium bicarbonate, dried with magnesium sulfate and concentrated under vacuum to yield 2-(5-bromo-thiazol-4-yl)-5-methyl-1H-benzoimidazole (1.93 g, crude, quantitative yield for two steps) as a beige solid that was used into the next step without further purification. 1H NMR (CHLOROFORM-d) δ: 8.82 (s, 1H), 7.64 (d, J=7.9 Hz, 1H), 7.47 (s, 1H), 7.15 (d, J=8.3 Hz, 1H), 2.50 (s, 3H); LCMS (EI/CI) m/z: 293.9, 295.9 [M+H]+.
4-(5-Methyl-1H-benzoimidazol-2-yl)-thiazole-5-carboxylic acid
A solution of 2-(5-bromo-thiazol-4-yl)-5-methyl-1H-benzoimidazole (0.48 g, 1.65 mmol) in anhydrous THF (20 mL) under argon atmosphere was cooled in a dry-ice/acetone bath. A solution of n-butyillithium (1.6M in hexanes, 2.27 mL, 3.63 mmol) was added drop-wise and the reaction continued for 30 min in the ice-bath. Then, a few pieces of dry ice were added to the reaction mixture and the mixture warmed to room temperature with stirring. The reaction was quenched by adding a solution of 1N HCl (20 mL) and the product was extracted with dichloromethane (4×30 mL). The combined organic layers were dried over anhydrous MgSO4 and concentrated under reduced pressure to give crude 4-(5-methyl-1H-benzoimidazol-2-yl)-thiazole-5-carboxylic acid (428 mg crude, ˜40%), as dark green solid, which was used directly for the next step without further purification. 1H NMR (METHANOL-d4) δ: 9.27-9.40 (m, 1H), 7.73-7.84 (m, 1H), 7.69 (s, 1H), 7.48 (d, J=8.3 Hz, 1H), 2.58 (d, J=2.3 Hz, 3H); LCMS (EI/CI) m/z: 257.9 [M−H]−.
To a solution of 4-(5-methyl-1H-benzoimidazol-2-yl)-thiazole-5-carboxylic acid (60 mg, 0.23 mmol) and HATU (149 mg, 0.39 mmol) in DMF (2 mL) was added tert-butylamine (0.1 mL, 0.92 mmol) and reaction stirred at room temperature for 8 h. The solvents were removed under high vacuum and the crude residue was purified by chromatography (silica, 0 to 5% of a 9:1 MeOH:ammonium hydroxide solution in dichloromethane, 20 min) to yield 4-(5-methyl-1H-benzoimidazol-2-yl)-thiazole-5-carboxylic acid tert-butylamide (0.072 g, 59%) as a white solid. 1H NMR (DMSO-d6) δ: 13.19 (br. s., 1H), 12.76 (s, 1H), 9.32 (s, 1H), 7.30-7.68 (m, 2H), 7.14 (br. s., 1H), 2.44 (s, 3H), 1.50 (s, 9H); LCMS (EI/CI) m/z: 315.0 [M+H]+.
N-[3-(1M-Benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide
2-(4-Nitro-1H-pyrazol-3-yl)-1H-benzoimidazole
A mixture of 4-nitro-1H-pyrazole-3-carboxylic acid (5.0 g, 31.8 mmol), 1,2-phenylenediamine (3.79 g, 35.0 mmol), EDC (7.30 g, 38.2 mmol) and HOBt (5.16 g, 38.2 mmol) in DMF (65 mL) was stirred at room temperature for 24 h. After which the solvent was reduced to a third of its original volume, and the remaining mixture was diluted with AcOH (60 mL) and refluxed for 3 h. Upon cooling, the solvents were removed under reduced pressure and water was added to the residue. The resultant precipitate was collected by filtration and washed thoroughly with water. The solid was dried through azeotroping with toluene to give 2-(4-nitro-1H-pyrazol-3-yl)-1H-benzoimidazole (5 g, 69%) as a yellow solid. LCMS (EI/CI) m/z: 228.0 [M+H]+.
3-(1H-Benzoimidazol-2-yl)-1H-pyrazol-4-ylamine
A mixture of 2-(4-nitro-1H-pyrazol-3-yl)-1H-benzoimidazole (3.5 g, 15.3 mmol) and 10% Pd—C (0.3 g) in DMF (40 mL) was subjected to an atmosphere of hydrogen (balloon pressure) at room temperature for 20 h. The mixture was then filtered through celite and the filtrate concentrated under reduced pressure. The residue was partitioned between ethyl acetate and water, the organic layer was separated, dried over anhydrous Na2SO4, and concentrated under reduced pressure to give crude 3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-ylamine (2.5 g crude, ˜82%) as a brown solid, which was used directly in the next step without further purification. LCMS (EI/CI) m/z: 200.2 [M+H]+.
N-[3-(1H-Benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide
To a solution of 3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-ylamine (300 mg, 1.51 mmol) in pyridine (8 mL), was added isobutyryl chloride (0.52 mL, 4.98 mmol) in dichloromethane (8 mL) at 0° C. The mixture was stirred at room temperature for 20 h, after which water was added and the mixture extracted with ethyl acetate (3×15 mL). The combined organic phases were washed with water, dried over anhydrous Na2SO4, and then concentrated under reduced pressure. The crude mass was redissolved in acetonitrile (5 mL) and methanol (10 mL) then 25% aqueous ammonium hydroxide solution (10 mL) was added to it. This mixture was stirred at r.t. for 3 h, then partitioned between EtOAc and water. The organic phase was collected and washed with 1N HCl. The organic phase was then poured over solid sodium bicarbonate, subsequently dried with anhydrous Na2SO4, and concentrated under reduced pressure. The crude mass was purified by chromatography (silica, EtOAc in hexanes) to give N-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide as off white solid (85 mg, 21%). LCMS (EI/CI) m/z: 269.8 [M+H]+.
N-[3-(1H-Benzoimidazol-2-yl)-1H-pyrazol-4-yl]-2,2-dimethyl-propionamide
N-[3-(1H-Benzoimidazol-2-yl)-1H-pyrazol-4-yl]-2,2-dimethyl-propionamide (180 mg, 42%) was synthesized as off white solid from 3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-ylamine (300 mg, 1.51 mmol) and pivaloyl chloride (0.61 mL, 4.98 mmol) following the procedure described for N-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide (Example 29). LCMS (EI/CI) m/z: 284.0 [M+H]+.
Cyclopentanecarboxylic acid [3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-amide
Cyclopentanecarboxylic acid [3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-amide (255 mg, 57%) was synthesized as white solid from 3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-ylamine (300 mg, 1.51 mmol) and cyclopentanecarbonyl chloride (0.63 mL, 4.98 mmol) following the procedure described for N-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide (Example 29). LCMS (EI/CI) m/z: 296.2 [M+H]+.
Tetrahydro-pyran-4-carboxylic acid [3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-amide
Tetrahydro-pyran-4-carboxylic acid [3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-amide (100.0 mg, 32%) was synthesized as light brown solid from 3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-ylamine (200 mg, 1.01 mmol) and tetrahydro-pyran-4-carbonyl chloride (494 mg, 4.98 mmol) following the procedure described for N-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide (Example 29). LCMS (EI/CI) m/z: 312.3 [M+H]+.
N-[3-(1H-Benzoimidazol-2-yl)-1H-pyrazol-4-yl]-3-hydroxy-2,2-dimethyl-propionamide
2-Acetoxymethyl-2-methyl-propionic acid
2,2-Dimethyl-3-hydroxypropionic acid (1.0 g, 8.48 mmol) was dissolved in pyridine (6 mL) at 0° C. and acetyl chloride (0.91 mL, 12.7 mmol) was added dropwise at this temperature. The mixture was warmed to room temperature and stirred for 4 h. After which a solution of 1NHCl was added to adjust the pH (to ˜3-4). The mixture was extracted with EtOAc (3×30 mL) and the combined organic layer dried over anhydrous Na2SO4. Concentration under reduced pressure then gave 2-acetoxymethyl-2-methyl-propionic acid (1.1 g, 81%) as an off-white crystalline solid which was used directly in the next reaction without further purification.
Acetic acid 2-chlorocarbonyl-2-methyl-propyl ester
A solution of 2-acetoxymethyl-2-methyl-propionic acid (2 g, 12.5 mmol) in benzene (20 mL) was cooled to 0° C. and oxalyl chloride (1.61 mL, 18.7 mmol) was added dropwise. The mixture was warmed to room temperature and stirred for 3 h. After which the mixture was concentrated under reduced pressure to give acetic acid 2-chlorocarbonyl-2-methyl-propyl ester as a yellow liquid (2.1 g crude, ˜94%). This material was used directly without further purification.
Acetic acid 2-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-ylcarbamoyl]-2-methyl-propyl ester
Acetic acid 2-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-ylcarbamoyl]-2-methyl-propyl ester (330 mg, 48%) was synthesized as light yellow solid from 3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-ylamine (400 mg, 2.1 mmol) and acetic acid 2-chlorocarbonyl-2-methyl-propyl ester (1.18 g, 6.63 mmol) following the procedure described for N-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-isobutyramide (Example 29). LCMS (EI/CI) m/z: 342.1 [M+H]+.
To a solution of acetic acid 2-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-ylcarbamoyl]-2-methyl-propyl ester (95 mg, 0.279 mmol) in MeOH (8 mL) was added K2CO3 (231 mg, 1.67 mmol). The mixture was stirred at room temperature for 2 h, then concentrated under reduced pressure. The residue obtained was diluted with ethyl acetate (25 mL), was washed with water (2×10 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude mass was then purified by chromatography (silica, EtOAc in hexanes) to give N-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-yl]-3-hydroxy-2,2-dimethyl-propionamide (72 mg, 86%) as a light yellow solid. LCMS (EI/CI) m/z: 300.2 [M+H]+.
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid isopropylamide
4-tert-Butoxy-3-oxo-butyric acid ethyl ester
To a solution of 2-methyl-propan-2-ol (25 g, 336 mmol) in DMF (250 mL) at 0° C. was added NaH (60%) (33.6 g, 841 mmol) in five portions. To this was slowly added 4-chloro-3-oxo-butyric acid ethyl ester (23.1 mL, 168 mmol) and the reaction mixture was stirred at 0° C. for 1 h and then warmed to room temperature and stirred for an additional 2 h. The mixture was then poured into ice water (200 mL) and extracted with EtOAc (3×100 mL). The organic extracts were combined and dried over anhydrous Na2SO4, concentrated under reduced pressure, and purified by chromatography (silica, EtOAc in hexanes) to give 4-tert-butoxy-3-oxo-butyric acid ethyl ester (14 g, 21%) as a sticky brown solid.
3-tert-Butoxymethyl-1H-pyrazole-4-carboxylic acid ethyl ester
To a solution of 4-tert-butoxy-3-oxo-butyric acid ethyl ester (14 g, 69 mmol) in toluene (100 mL) was added dimethylformamide dimethyl acetal (14 mL). The mixture heated at 65° C. for 3 h. Upon cooling, the volatiles were removed under reduced pressure and the residue was then redissolved in acetic acid (20 mL), and hydrazine hydrate (5.06 mL, 104 mmol) was added. After stirring at room temperature for 3 h, the reaction mixture was poured into water (100 mL) and extracted with EtOAc (3×75 mL). The combined organic phases were dried over anhydrous Na2SO4, concentrated under reduced pressure and the crude mass was purified by chromatography (silica, EtOAc/hexanes) to give 3-tert-butoxymethyl-1H-pyrazole-4-carboxylic acid ethyl ester (6.5 g, 41%) as a brown liquid. LCMS (EI/CI) m/z: 227.2 [M+H]+.
3-Hydroxymethyl-1H-pyrazole-4-carboxylic acid ethyl ester
To a solution of 3-tert-butoxymethyl-1H-pyrazole-4-carboxylic acid ethyl ester (6.5 g, 28.8 mmol) in dichloromethane (100 mL) was added trifluoroactetic acid (50 mL). The mixture was then stirred at room temperature for 2 h and then concentrated in vacuo. The crude residue was poured into water (50 mL), neutralized with saturated aqueous sodium bicarbonate, and then extracted with EtOAc (3×50 mL). The combined organic extracts were dried over anhydrous Na2SO4 and concentrated under reduced pressure to give 3-hydroxymethyl-1H-pyrazole-4-carboxylic acid ethyl ester (5.3 g) as light brown liquid which was used directly in next step without further purification. LCMS (EI/CI) m/z: 171.2 [M+H]+.
3-Formyl-1H-pyrazole-4-carboxylic acid ethyl ester
To a stirred solution of 3-hydroxymethyl-1H-pyrazole-4-carboxylic acid ethyl ester (4.5 g, 26.5 mmol) in EtOAc (50 mL) was added IBX (22.2 g, 79.4 mmol). The reaction mixture was heated to 60° C. for 6 h. Upon cooling, the mixture was filtered through a bed of celite and the filtrate was then concentrated under reduced pressure. The crude mass was purified by chromatography (silica, EtOAc in hexanes) to give 3-formyl-1H-pyrazole-4-carboxylic acid ethyl ester (2.0 g, 45%) as a gum. This semi-pure material was used directly in the next step without further purification.
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester
To a solution of 3-formyl-1H-pyrazole-4-carboxylic acid ethyl ester (2.0 g, 11.9 mmol) and benzene-1,2-diamine (2.3 g, 21.4 mmol) in DMF (20 mL) was added NaHSO3 (4.2 g, 40.4 mmol). The reaction mixture was heated at 120° C. for 3 h. Upon cooling the mixture was poured onto crushed ice and the precipitate was collected by filtration. This solid was washed with saturated sodium bicarbonate followed by water, then dried under reduced pressure to give 3-(1H-benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (1.0 g, 33%) as a brown solid. LCMS (EI/CI) m/z: 257.2 [M+H]+.
To a mixture of (3-(1H-benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (200 mg, 0.391 mmol) and isopropyl amine (0.09 mL, 1.95 mmol) was added Me3Al (2 M in THF, 0.19 mL, 0.391 mmol) under and argon atmosphere. The mixture was heated at 60° C. for 16 h, after which, the mixture was cooled, diluted with water (10 mL) and extracted with EtOAc (3×15 mL). The combined organic extracts were dried over anhydrous Na2SO4 and concentrated under reduced pressure. The crude residue was purified by preparative HPLC (NH4OAc-CH3CN solvent mixture) to give 3-(1H-benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid isopropylamide (45 mg, 43%) as an off-white solid. LCMS (EI/CI) m/z: 270.4 [M+H]+.
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid tert-butylamide
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid tert-butylamide (42 mg, 19%) was synthesized as an off-white solid from 3-(1H-benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (200 mg, 0.781 mmol) and tert-butyl amine (0.41 mL, 3.91 mmol) following the procedure described for 3-(1H-benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid isopropylamide (Example 35). LCMS (EI/CI) m/z: 284.0 [M+H]+.
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid (tetrahydro-pyran-4-yl)-amide
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid (tetrahydro-pyran-4-yl)-amide (30 mg, 16%) was synthesized as an off white solid from 3-(1H-benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (150 mg, 0.586 mmol) and tetrahydro-pyran-4-ylamine (118 mg, 1.17 mmol) following the procedure described for 3-(1H-benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid isopropylamide (Example 35). LCMS (EI/CI) m/z: 312.2 [M+H]+.
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid (2-hydroxy-1,1-dimethyl-ethyl)-amide
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid (2-hydroxy-1,1-dimethyl-ethyl)-amide (60 mg, 26%) was synthesized as a white solid from 3-(1H-benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (200 mg, 0.781 mmol) and 2-amino-2-methyl-propan-1-ol (344 mg, 3.91 mmol) following the procedure described for 3-(1H-benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid isopropylamide (Example 35). LCMS (EI/CI) m/z: 300.2 [M+H]+.
[4-(4-Methyl-1H-benzoimidazol-2-yl)-thiazol-5-yl]-pyridin-3-yl-amine
5-(Pyridin-3-ylamino)-thiazole-4-carboxylic acid ethyl ester
A solution of t-BuOK (5.45 g, 48.6 mmol) in THF (25 mL) was cooled to −40° C. To this mixture was added drop-wise, keeping the temperature below −35° C., a solution of isocyano-acetic acid ethyl ester (5 g, 44.2 mmol) and 3-isothiocyanato-pyridine (6.02 g, 44.2 mmol) in THF (50 mL). After the addition was complete, the reaction was warmed to −9° C. over 1 h. The reaction was then quenched by the addition of 2.5 mL of acetic acid. The reaction mixture was diluted with THF and ethyl acetate, filtered through a pad of celite and the filtrates evaporated. The residue was purified by chromatography (silica, 50% to 100% ethyl acetate in heptane, 30 min) to give 5-(pyridin-3-ylamino)-thiazole-4-carboxylic acid ethyl ester (4.85 g, 44%). 1H NMR (CHLOROFORM-d) δ: 9.86 (br. s., 1H), 8.62 (d, J=3.0 Hz, 1H), 8.37 (dd, J=4.7, 1.2 Hz, 1H), 8.05 (d, J=1.1 Hz, 1H), 7.60 (ddd, J=8.2, 2.8, 1.3 Hz, 1H), 7.33 (dd, J=8.3, 4.6 Hz, 1H), 4.47 (q, J=7.0 Hz, 2H), 1.47 (t, J=7.1 Hz, 3H); MS (EI/CI) m/z: 250.2 [M+H]+.
5-(Pyridin-3-ylamino)-thiazole-4-carboxylic acid
A solution of 5-(pyridin-3-ylamino)-thiazole-4-carboxylic acid ethyl ester (1.56 g, 6.25 mmol) in methanol (12 mL) was treated with a solution of KOH (1.05 g, 18.7 mmol) in water (9 mL). The mixture was heated to 65° C. with stirring for 3 h. The methanol was evaporated, and the residual aqueous solution was then acidified to pH 4 with 1 N HCl. The gel that formed was treated with some methanol and the suspension sonicated until a loose off-white suspension was formed. The suspension was separated by filtration, washed with water, ethyl ether and then dried under high vacuum to give 5-(pyridin-3-ylamino)-thiazole-4-carboxylic acid (1.24 g, 90%) as an off-white solid. 1H NMR (DMSO-d6) δ: 12.79 (br. s., 1H), 9.73 (s, 1H), 8.63 (d, J=2.7 Hz, 1H), 8.26-8.35 (m, 2H), 7.79 (ddd, J=8.3, 2.8, 1.3 Hz, 1H), 7.42 (dd, J=8.3, 4.6 Hz, 1H); MS (EI/CI) m/z: 220.2 [M−H]−.
A solution of 5-(pyridin-3-ylamino)-thiazole-4-carboxylic acid (0.2 g, 0.90 mmol), diisopropylethylamine (0.24 mL, 1.17 mmol) and 3-methyl-benzene-1,2-diamine (0.122 g, 0.99 mmol) in DMF (3 mL) at room temperature was treated with TBTU (0.38 g, 1.17 mmol) and the mixture stirred for 18 h at room temperature. The solvents were evaporated under vacuum and residue obtained was then partitioned between ethyl acetate and a saturated solution of sodium bicarbonate. The organic phase was dried with magnesium sulfate and concentrated under vacuum. The crude mixture was dissolved in acetic acid (3 mL), transferred to microwave vial and the mixture was heated at 120° C. in a microwave reactor for 2 h. The mixture was cooled, concentrated in vacuo, and the residue then dissolved in dichloromethane, washed with saturated solution of sodium bicarbonate, dried with magnesium sulfate and concentrated under vacuum. The residue was purified by chromatography (silica, 0 to 5% methanol in dichloromethane) to yield [4-(4-methyl-1H-benzoimidazol-2-yl)-thiazol-5-yl]-pyridin-3-yl-amine (0.038 g, 14% over two steps) as a beige solid. 1H NMR (CHLOROFORM-d) δ: 8.73 (d, J=2.6 Hz, 1H), 8.32 (dd, J=4.5, 1.1 Hz, 1H), 8.20 (s, 1H), 7.69 (d, J=7.6 Hz, 1H), 7.47 (br. s., 1H), 7.36 (m, 2H), 7.22 (m, 2H), 7.11 (d, J=7.2 Hz, 1H), 2.68 (s, 3H); MS (EI/CI) m/z: 308.0 [M+H]+.
[4-(1H-Imidazo[4,5-c]pyridin-2-yl)-thiazol-5-yl]-pyridin-3-yl-amine
A solution of 5-(pyridin-3-ylamino)-thiazole-4-carboxylic acid (0.2 g, 0.90 mmol), diisopropylethylamine (0.24 mL, 1.17 mmol) and pyridine-3,4-diamine (0.108 g, 0.99 mmol) in DMF (3 mL) at room temperature was treated with TBTU (0.38 g, 1.17 mmol) and the mixture stirred for 18 h at room temperature. The mixture was concentrated under vacuum and the residue dissolved in ethyl acetate, washed with a saturated solution of sodium bicarbonate, and then dried with magnesium sulfate and concentrated under vacuum. The crude mixture was dissolved in acetic acid (3 mL), transferred to microwave vial and mixture was heated in a microwave reactor at 120° C. for 4.5 h, then at 150° C. for 1.5 h. The mixture was cooled, concentrated, and the residue obtained was then dissolved in dichloromethane, washed with saturated solution of sodium bicarbonate, dried with magnesium sulfate and concentrated under vacuum. The residue was purified by chromatography (silica, 0 to 10% methanol in dichloromethane, 20 min) to give [4-(1H-imidazo[4,5-c]pyridin-2-yl)-thiazol-5-yl]-pyridin-3-yl-amine (0.016 g, 6% over two steps) as an off-white solid. 1H NMR (DMSO-d6) δ: 8.98 (s, 1H), 8.79 (s, 1H), 8.63-8.74 (m, 2H), 8.23-8.38 (m, 2H), 7.81 (dd, J=7.7, 2.5 Hz, 1H), 7.70 (br. s., 1H), 7.40-7.52 (m, 2H); MS (EI/CI) m/z: 295.0 [M+H]+.
(1,1-Dimethyl-propyl)-[4-(5-methyl-1H-benzoimidazol-2-yl)-thiazol-5-yl]-amine
5-(1,1-Dimethyl-propylamino)-thiazole-4-carboxylic acid ethyl ester
Prepared in a parallel fashion using the procedure described for example 39; MS (EI/CI) m/z: 243.6 [M+H]+.
5-(1,1-Dimethyl-propylamino)-thiazole-4-carboxylic acid
Prepared in a parallel fashion using the procedure described for example 39; MS (EI/CI) m/z: 213.0 [M−H]−.
A solution of 5-(1,1-dimethyl-propylamino)-thiazole-4-carboxylic acid (0.153 g, 0.71 mmol), diisopropylethylamine (0.16 mL, 0.93 mmol) and 4-methyl-benzene-1,2-diamine (0.096 g, 0.78 mmol) in DMF (2 mL) at room temperature was treated with TBTU (0.3 g, 0.93 mmol) and the mixture stirred for 18 h at room temperature. The mixture was then concentrated under vacuum and the residue dissolved ethyl acetate, washed with a saturated solution of sodium bicarbonate, dried with magnesium sulfate and then concentrated under vacuum. The crude mixture was dissolved in acetic acid (3 mL), transferred to microwave vial and heated in a microwave reactor at 120° C. for 2 h. The mixture was cooled, concentrated in vacuo, then the residue was dissolved in dichloromethane, washed with a saturated solution of sodium bicarbonate, dried with magnesium sulfate and concentrated under vacuum. The residue was purified by chromatography (silica, 0 to 5% of a 9:1 methanol:ammonium hydroxide solution in dichloromethane, 20 min) to yield (1,1-dimethyl-propyl)-[4-(5-methyl-1H-benzoimidazol-2-yl)-thiazol-5-yl]-amine (0.044 g, 21%) as an dark green oil. 1H NMR (CHLOROFORM-d) δ: 10.48-10.81 (m, 1H), 8.73 (d, J=17.4 Hz, 1H), 7.99 (s, 1H), 7.46-7.67 (m, 1H), 7.20-7.35 (m, 1H), 6.94-7.11 (m, 1H), 2.48 (d, J=4.5 Hz, 3H), 1.82 (q, J=7.3 Hz, 2H), 1.46 (s, 6H), 1.00 (t, J=7.4 Hz, 3H); MS (EI/CI) m/z: 301.0 [M+H]+.
Cyclopentyl-[4-(5-methyl-1H-benzoimidazol-2-yl)-thiazol-5-yl]-amine
5-Cyclopentylamino-thiazole-4-carboxylic acid ethyl ester
Prepared in a parallel fashion using the procedure described for example 39; MS (EI/CI) m/z: 241.7 [M+H]+.
5-Cyclopentylamino-thiazole-4-carboxylic acid
Prepared in a parallel fashion using the procedure described for example 39; MS (EI/CI) m/z: 213.4 [M+H]+.
A solution of 5-cyclopentylamino-thiazole-4-carboxylic acid (0.124 g, 0.58 mmol), diisopropylethylamine (0.13 mL, 0.76 mmol) and 4-methyl-benzene-1,2-diamine (0.078 g, 0.64 mmol) in DMF (2 mL) at room temperature was treated with TBTU (0.244 g, 0.76 mmol) and the mixture stirred for 18 h at room temperature. The solvents were removed under vacuum and the residue then dissolved in ethyl acetate, washed with a saturated solution of sodium bicarbonate, dried over magnesium sulfate and finally concentrated under vacuum. The crude mixture was dissolved in acetic acid (3 mL), transferred to microwave vial and heated at 120° C. in a microwave reactor for 2 h. The mixture was cooled, concentrated in vacuo, and then dissolved in dichloromethane. The organic phase was washed with a saturated solution of sodium bicarbonate, dried with magnesium sulfate and concentrated under vacuum. The residue was purified by chromatography (silica, dichloromethane) to yield cyclopentyl-[4-(5-methyl-1H-benzoimidazol-2-yl)-thiazol-5-yl]-amine (0.035 g, 20%) as a beige solid. 1H NMR (CHLOROFORM-d) δ: 10.35-10.58 (m, 1H), 8.16 (dd, J=17.6, 6.6 Hz, 1H), 7.98 (s, 1H), 7.47-7.63 (m, 1H), 7.19-7.33 (m, 1H), 6.95-7.09 (m, 1H), 3.65-3.86 (m, 1H), 2.48 (d, J=4.2 Hz, 3H), 2.00-2.24 (m, 2H), 1.59-1.96 (m, 6H); MS (EI/CI) in/z: 299.1 [M+H]+.
Isobutyl-[4-(5-methyl-1H-benzoimidazol-2-yl)-thiazol-5-yl]-amine
5-Isobutylamino-thiazole-4-carboxylic acid ethyl ester
Prepared in a parallel fashion using the procedure described for example 39; MS (EI/CI) m/z: 229.7 [M+H]+.
5-Isobutylamino-thiazole-4-carboxylic acid
Prepared in a parallel fashion using the procedure described for example 39; MS (EI/CI) m/z: 201.7 [M+H]+.
A solution of 5-isobutylamino-thiazole-4-carboxylic acid (0.160 g, 0.80 mmol), diisopropylethylamine (0.18 mL, 1.04 mmol) and 4-methyl-benzene-1,2-diamine (0.107 g, 0.88 mmol) in DMF (2 mL) at room temperature was treated with TBTU (0.333 g, 1.04 mmol) and mixture stirred for 18 h at room temperature. The mixture was concentrated in vacuo and the residue obtained dissolved in ethyl acetate. The organic solution was washed with a saturated solution of sodium bicarbonate, dried with magnesium sulfate and then concentrated under vacuum. The crude mixture was dissolved in acetic acid (3 mL), transferred to microwave vial and heated in a microwave reactor at 120° C. for 2 h. The mixture was cooled, concentrated in vacuo, and then the residue was dissolved in dichloromethane. This solution was washed with a saturated solution of sodium bicarbonate, dried with magnesium sulfate and concentrated under vacuum. The residue was purified by chromatography (silica, dichloromethane) to give isobutyl-[4-(5-methyl-1H-benzoimidazol-2-yl)-thiazol-5-yl]-amine (0.066 g, 29%) as a dark red oil. 1H NMR (CHLOROFORM-d) δ: 10.43-10.62 (m, 1H), 8.30 (dt, J=16.1, 5.8 Hz, 1H), 7.97 (s, 1H), 7.51 (s, 1H), 7.18-7.34 (m, 1H), 6.94-7.09 (m, 1H), 3.19 (t, J=6.2 Hz, 2H), 2.48 (d, J=4.5 Hz, 3H), 2.10 (dquin, J=13.5, 6.6 Hz, 1H), 1.08 (d, J=6.8 Hz, 6H); MS (EI/CI) in/z: 287.1 [M+H]+.
SYK Assay Information
Determination of IC50 of Spleen Tyrosine Kinase (SYK) Inhibition:
SYK kinase assay is a standard kinase assay adapted to a 96 well plate format. This assay is performed in 96-well format for IC50 determination with 8 samples which represented 10 half log dilutions and a 40 μL reaction volume. The assay measures the incorporation of radiolabeled 33P ATP into an N-terminally biotinylated peptide substrate, derived from naturally occurring phosphoacceptor consensus sequence (Biotin-11aa DY*E). Phosphorylated products were detected upon termination of reactions with EDTA and the addition of Streptavidin coated beads. Representative results are in Table II above.
Assay plates: 96-well MultiScreen 0.65 um filter plates (Millipore Cat. No.: MADVNOB10) Streptavidin coated beads: Streptavidin Sepharose™, suspension 5.0 mL, in 50 mM EDTA/PBS diluted (1:100), (Amersham, Cat. No.: 17-5113-01)
Compounds: 10 mM in 100% dimethylsulfoxide (DMSO), final conc.: compound 0.003-100 uM in 10% DMSO
Enzyme: SYK RPA purified, truncated construct of Spleen Tyrosine Kinase aa 360-635, stock solution 1 mg/mL, MW: 31.2 KDa, final conc.:0.0005 μM.
Peptide 1: biotinylated peptide is derived from a naturally occurring phosphor-acceptor consensus sequence (Biotin-EPEGDYEEVLE), special order from QCB, stock solution 20 mM, final conc.: 5.0 μM.
ATP: Adenosine-5′-triphosphate 20 mM, (ROCHE Cat. No.: 93202720), final concentration: 20 μM
Buffer: HEPES: 2-Hydroxyethyl piperazine-2-ethanesulfonic acid (Sigma, Cat. No.: H-3375) final concentration: 50 mM HEPES pH7.5
BSA: Bovine Serum Albumin Fraction V, fatty acid free (Roche Diagnostics GmbH, Cat. No. 9100221) diluted to a final concentration of 0.1%
EDTA: EDTA stock solution 500 mM, (GIBCO, Cat. No.: 15575-038) final concentration: 0.1 mM
DTT: 1,4-Dithiothreitol (Roche Diagnostics GmbH, Cat. No.: 197777), final conc.: 1 mM MgCl2×6H2O: MERCK, Cat. No.: 105833.1000, final concentration: 10 mM
Assay Dilution Buffer (ADB): 50 mM HEPES, 0.1 mM EGTA, 0.1 mM Na Vanadate, 0.1 mM β-glycerophosphate, 10 mM MgCl2, 1 mM DTT, 0.1% BSA, pH 7.5
Bead wash buffer: 10 g/L PBS (Phosphate buffered saline) with 2M NaCl+1% phosphoric acid.
In 40 μL volume, 26 μL of ADB diluted, purified recombinant human SYK360-635 [0.5 nM] was mixed with 4 μL of 10× concentrations of the test compounds, [usually 100 μM-0.003 μM] in [10%] DMSO and the mixture was incubated for 10 min at RT.
The kinase reaction was initiated by the addition of 10 μL 4× substrate cocktail containing the DYE peptide substrate [0 or 5 μM], ATP [20 μM] and 33PγATP [2 μCi/rxn]. After incubation at 30° C. for 15 min, the reaction was terminated by the transfer of 25 μL pf the reaction sample to a 96 well 0.65 μm Millipore MADVNOB membrane/plate containing 200 μL 5 mM EDTA and 20% Streptavidine coated beads in PBS.
The unbound radionucleotides were washed under vacuum with 3×250 μL 2M NaCl; 2×250 μL 2M NaCl+1% phosphoric acid; 1×250 μL H2O. After the last wash membrane/plates were transferred to an adaptor plate, heat dried for 15 min at 60° C., and 50 μL scintillation cocktail was added to each well and 4 h later the amount of radioactivity was counted in a top counter.
The percent inhibition was calculated based on the uninhibited enzyme rate:
% Inhibition=100/(1+(IC50/Inhibitor conc)n)
The IC50 was calculated using a non-linear curve fit with XLfit software (ID Business Solution Ltd., Guilford, Surrey, UK).
IC50: human Compound ENZYME_FILTRATION_IC50 RAMOS_V2_IC50 whole blood
I-1 35.1892 I-2 0.1126 >34.09 I-3 2.0773 5.40966 I-4 0.6209 3.831025 I-5 1.27905 4.645375 I-6 2.75945 11.98022 I-7 0.5619 0.551606 20.3658 I-8 0.067 0.145682 46.6636 I-9 >10 >50 I-10 >10 >50 I-11 7.94955 >34.09 I-12 1.687 4.07351 13.1175 I-13 >10 >34.09 >50 I-14 >10 >34.09 I-15 >10 >34.09 I-16 2.40935 12.160015 >50 I-17 0.7608 3.5410475 >50 I-18 6.1238 >34.09 >50 I-19 >34.09 >50 I-20 0.2154 >50 I-21 0.20895 19.067255 >50 I-22 1.99345 >34.09 I-23 7.87075 >34.09 I-24 >50 I-25 4.60585 5.20907 I-26 1.22925 I-27 4.64035 15.507915 I-28 1.12665 6.412955 I-29 0.0466 1.520248 10.3825 I-30 0.06005 1.016505 2.2871 I-31 0.02145 0.349586667 2.6025 I-32 0.0389 0.61519 5.2724 I-33 0.1048 3.08306 2.5008 I-34 0.27845 6.568568 4.1701 I-35 0.05495 0.92046 1.9186 I-36 0.292265 2.5424 I-37 0.140125 2.8606 I-38 0.483085 0.87 I-39 0.9864 2.8688 I-40 0.4872 0.7637985 I-41 5.43715 >31.25 I-42 1.6235 >50 I-43 0.69885 >34.09 I-44 0.62125 7.984055
1. A compound, selected from the group consisting of:
N-[3-(1H-Benzoimidazol-2-yl)-1H-pyrazol-4-yl]-2,2-dimetyl-propionamide;
Acetic acid 2-[3-(1H-benzoimidazol-2-yl)-1H-pyrazol-4-ylcarhamoyl]-2-methyl-propyl ester;
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid isopropylamide;
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid tert-butylamide; and
3-(1H-Benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid (2-hydroxy-1,1-dimethyl-ethyl) amide
6897208 May 24, 2005 Edwards
7179823 February 20, 2007 Momose et al.
7977477 July 12, 2011 Berdini
8399442 March 19, 2013 Berdini et al.
20090131470 May 21, 2009 Walmsley et al.
20100160324 June 24, 2010 Berdini et al.
2002161084 June 2002 JP
2007119484 May 2007 JP
03066629 August 2003 WO
2005002576 January 2005 WO
2005005414 January 2005 WO
2006070192 July 2006 WO
2006091592 August 2006 WO
2008129054 October 2008 WO
2009027393 March 2009 WO
2010091310 August 2010 WO
2011003065 January 2011 WO
2011150457 December 2011 WO
2012025186 March 2012 WO
2013124025 August 2013 WO
2014041175 March 2014 WO
The International Search Report and Written Opinion, issued on Nov. 15, 2013, in the corresponding PCT Application No. PCT/EP2013/072132.
Ribecai et al., “Identification of a Manufacturing Route of Novel CRF-1 Antagonists Containing a 2,3-Dihydro-1H-pyrrolo[2,3-b]pyridine Moiety,” Org. Process Res. Dev., 2010, 14 (4), pp. 895-901.
Howard et al., “Fragment-based discovery of the pyrazol-4-yl urea (AT9283), a multitargeted kinase inhibitor with potent aurora kinase activity,” J Med Chem. Jan. 22, 2009;52(2):379-88.
Schiffmann et al., “Metal-mediated inhibition of Escherichia coli methionine aminopeptidase: structure-activity relationships and development of a novel scoring function for metal-ligand interactions,” J Med Chem. Jan. 26, 2006;49 (2):511-22.
Tocco et al.,“The Metabolic Fate of Thiabendazole in Sheep,” J Med Chem. Jul. 1964;7:399-405.
Amoroso et al., “Synthesis of the new tripodal ligand tris-[3-(2′-pyridyl)pyrazol-1-yl]hydroborate, and the crystal structure of its europium(III) complex,” J Chem Soc, Chem Commun, 1994, p. 2751-2752.
Plate et al., “Synthesis and muscarinic activities of 3-(pyrazolyl)-1,2,5,6-tetrahydropyridine derivatives,” Bioorg Med Chem. Feb. 1996;4(2):227-37.
Hayat et al., “Synthesis, characterization, antiamoebic activity and cytotoxicity of novel 2-(quinolin-8-yloxy) acetohydrazones and their cyclized products (1,2,3-thiadiazole and 1,2,3-selenadiazole derivatives),” Eur J Med Chem. Dec. 2010;45(12):6127-34.
Feiyue et al., “Bidentate Ligands That Contain Pyrrole in Place of Pyridine,” Inorg. Chem., 2000, 39 (3), pp. 584-590.
The English translation of the Chinese Office Action, dated Apr. 8, 2016, in the corresponding Chinese Application No. 201380042831.9.
Hohwy et al., “Novel Prostaglandin D Synthase Inhibitors Generated by Fragment-Based Drug Design,” J. Med. Chem. 2008, 51, 2178-2186.
The English translation of the Taiwanese Search Report, dated Mar. 23, 2017, in the corresponding Taiwanese Application No. 102138784.
The English translation of the Chinese Office Action, dated Mar. 21, 2017, in the related Chinese Application No. 01380042831.9.
The English translation of the Japanese Office Action, dated Jul. 25, 2017, in the related Japanese Application No. 2015-538425.
The English translation of the Russian Office Action, dated Aug. 13, 2017, in the related Russian Application No. 2015117950.
Isaburo et al., “The Paal-Knorr Conden6ation of Acetonyincetone with 5-Amin opyrazoles,” Bull. Chem. Soc. Jpn. 1971, vol. 44, pp. 2856-2858.
Patent number: 9988378
Patent Publication Number: 20150266874
Assignee: Hoffmann-La Roche Inc. (Basle)
Inventors: Niala Bhagirath (Bloomfield, NJ), Joshua Kennedy-Smith (New York, NY), Matthew C. Lucas (Lexington, MA), Fernando Padilla (Verona, NJ)
Application Number: 14/437,307
International Classification: C07D 403/04 (20060101); C07D 471/04 (20060101); C07D 401/04 (20060101); C07D 403/14 (20060101); C07D 409/04 (20060101); C07D 417/04 (20060101); C07D 417/14 (20060101); A61K 31/415 (20060101); A61K 31/4155 (20060101); A61K 31/427 (20060101); A61K 31/428 (20060101); A61K 31/437 (20060101); A61K 31/4439 (20060101); A61K 31/454 (20060101); A61K 45/06 (20060101);