Patent Application
20250074913
The present invention relates to organic compounds, in particular to STING agonists, useful for treatment of cancers in a mammal. In particular, the present invention relates to pyrazole derivatives that have STING agonism activity, as well as their manufacture, pharmaceutical compositions containing them and their potential use as medicaments.
Cancer immunotherapy (CIT), represented by immune checkpoint inhibitors (ICI), have rapidly emerged in the past few years. The key to an effective CIT-mediated antitumor responses depend on the priming and activation of T cells by antigen-presenting cells, followed by directed trafficking and infiltration of T cells into the tumor mass so that they recognize and kill cancer cells (Chen D S, et al. Immunity. 2013; 39). As such, immune checkpoint blockade only shows durable responses in highly immunogenic “hot” tumors but has shown little efficacy in non-immunogenic “cold” tumors which are characterized by the lack of T-cell infiltration and impaired T-cell priming. Therefore, attempts have been focused on promoting the transformation of “cold tumors” into “hot tumors”, with strategies to activate the innate immune system to re-engage non-immunogenic tumors by boosting anti-cancer adaptive immunity. Antigen presentation cells of the innate immune system, such as dendritic cells or macrophages, serve as a critical link between the innate and adaptive immune systems by phagocytosis to process the foreign antigens and present them on the cell surface to the T cells, thereby activating T cell response.
Stimulator of interferon genes (STING) is a four transmembrane endoplasmic reticulum adaptor that facilitates innate immune signaling. Activation of STING protein by its natural ligand, cyclic guanosine monophosphate adenosine monophosphate (cGAMP) is able to activate NF-kB, IRF3 transcription pathway to induce expression of type I interferon and pro-inflammatory cytokine/chemokines (Ishikawa. et al. Nature 2008, Quyang et al. Immunity 2012, Chen et al. Cell 2011)
STING activation has been demonstrated to enhance the immune response and restrain tumor growth, which is achieved by strengthen cancer antigen presentation, contribute to the priming and activation of T cells, facilitate the trafficking and infiltration of T cells into tumors and promote the recognition and killing of cancer cells by T cells. The first generation of intra-tumor delivered cyclic dinucleotides that have entered clinical development only demonstrated limited single agent activity, mainly due to the poor pharmacokinetic and physicochemical properties, i.e., low bioavailability, instability, and difficulty in bypassing the plasma membrane (Leila et al. J. Clin. Med. 2020, Afsaneh et al. Cancers 2021).
Objects of the present invention are novel compounds of formula (I), their manufacture, medicaments based on a compound in accordance with the invention and their production as well as the use of compounds of formula (I) as STING agonists for the treatment of a broad spectrum of cancers, with the potential to augment response rates to current immunotherapy, overcome acquired resistance. The compounds of formula (I) show superior STING agonism activity. In addition, the compounds of formula (I) also show good safety and good PK profiles, e.g., good solubility, microsome stability, permeability, and safety margin.
One aspect of the invention pertains to a compound of formula (I),
Another aspect of the invention pertains to a process for the preparation of a compound of formula (I), as well as a compound of formula (I) or a pharmaceutically acceptable salt thereof when manufactured according to the process.
Another aspect of the invention pertains to a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.
Another aspect of the invention pertains to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use as therapeutically active substance.
Another aspect of the invention pertains to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer.
Another aspect of the invention pertains to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof for the treatment of cancer.
Another aspect of the invention pertains to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof for the activation of STING.
Another aspect of the invention pertains to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of cancer.
Another aspect of the invention pertains to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the activation of STING.
Another aspect of the invention pertains to a method for the treatment of cancer, which method comprises administering an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Furthermore, the following definitions are set forth to illustrate and define the meaning and scope of the various terms used to describe the invention.
The nomenclature used in this application is based on IUPAC systematic nomenclature, unless indicated otherwise.
The term “compound(s) of this invention” and “compound(s) of the present invention” refers to compounds of formula (I), formula (Ia), formula (Ib), and stereoisomers, solvates or salts thereof (e.g., pharmaceutically acceptable salts).
The term “substituent” denotes an atom or a group of atoms replacing a hydrogen atom on the parent molecule.
As used herein, the term “C1-6alkyl” alone or in combination signifies a saturated, linear- or branched chain alkyl group containing 1 to 6, particularly 2 to 6 or 1 to 4 carbon atoms, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl and the like. Particular “C1-6alkyl” groups are methyl and ethyl.
The term “halogen” or “Halo” denotes fluoro, chloro, bromo, or iodo.
The term “haloC1-6alkyl” denotes an alkyl group wherein at least one of the hydrogen atoms of the alkyl group is replaced by same or different halogen atoms, particularly fluoro atoms. Examples of haloC1-6alkyl include monochloro-, difluoro-or trifluoro-methyl, -ethyl or -propyl, for example difluoromethyl.
The term “heteroaryl” denotes a monovalent aromatic heterocyclic mono- or bicyclic ring system of 5 to 12 ring atoms, comprising 1, 2, 3 or 4 heteroatoms selected from N, O and S, the remaining ring atoms being carbon. Examples of heteroaryl moieties include, but not limited to, pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, thiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl, triazinyl, isoxazolyl, benzofuranyl, isothiazolyl, benzothienyl, indolyl, isoindolyl, isobenzofuranyl, benzimidazolyl, benzoxazolyl, benzoisoxazolyl, benzothiazolyl, benzoisothiazolyl, benzooxadiazolyl, benzothiadiazolyl, benzotriazolyl, purinyl, quinolinyl, isoquinolinyl, quinazolinyl or quinoxalinyl. Heteroaryl can be further substituted by halogen, C1-6alkyl, haloC1-6alkyl, cyano, C3-7cycloalkyl, (C1-6alkyl)2amino, or C1-6alkoxy.
The term “heterocyclyl” denotes a monovalent saturated or partly unsaturated mono- or bicyclic ring system of 3 to 9 ring atoms, comprising 1, 2, or 3 ring heteroatoms selected from N, O and S, the remaining ring atoms being carbon. In particular embodiments, heterocyclyl is a monovalent saturated monocyclic ring system of 4 to 7 ring atoms, comprising 1, 2, or 3 ring heteroatoms selected from N, O and S, the remaining ring atoms being carbon. Examples for monocyclic saturated heterocyclyl are aziridinyl, oxiranyl, azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydro-thienyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholin-4-yl, azepanyl, diazepanyl, homopiperazinyl, oxazepanyl, oxopiperidinyl, oxopiperazinyl or oxopyrrolidinyl. Examples for bicyclic saturated heterocyclyl are azaspiro[3.3]heptanyl, 8-aza-bicyclo[3.2.1]octyl, quinuclidinyl, 8-oxa-3-aza-bicyclo[3.2.1]octyl, 9-aza-bicyclo[3.3.1]nonyl, 3-oxa-9-aza-bicyclo[3.3.1]nonyl, or 3-thia-9-aza-bicyclo[3.3.1]nonyl. Examples for partly unsaturated heterocyclyl are dihydrofuryl, imidazolinyl, dihydro-oxazolyl, tetrahydro-pyridinyl, or dihydropyranyl.
The term “hydroxyC1-6alkyl” denotes a C1-6alkyl group wherein at least one of the hydrogen atoms of the C1-6alkyl group has been replaced by a hydroxy group.
The compounds according to the present invention may exist in the form of their pharmaceutically acceptable salts. The term “pharmaceutically acceptable salt” refers to conventional acid-addition salts or base-addition salts that retain the biological effectiveness and properties of the compounds of formula (I) and are formed from suitable non-toxic organic or inorganic acids or organic or inorganic bases. Acid-addition salts include for example those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid, and those derived from organic acids such as p-toluenesulfonic acid, trifluoroacetic acid, formic acid, salicylic acid, methanesulfonic acid, oxalic acid, succinic acid, citric acid, malic acid, lactic acid, fumaric acid, and the like. Base-addition salts include those derived from ammonium, potassium, sodium and, quaternary ammonium hydroxides, such as for example, tetramethyl ammonium hydroxide. The chemical modification of a pharmaceutical compound into a salt is a technique well known to pharmaceutical chemists in order to obtain improved physical and chemical stability, hygroscopicity, flowability and solubility of compounds. It is for example described in Bastin R. J., et al., Organic Process Research & Development 2000, 4, 427-435. Particular are the sodium salts of the compounds of formula (I).
The term “therapeutically effective amount” denotes an amount of a compound or molecule of the present invention that, when administered to a subject, (i) treats or prevents the particular disease, condition or disorder, (ii) attenuates, ameliorates or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, condition or disorder described herein. The therapeutically effective amount will vary depending on the compound, the disease state being treated, the severity of the disease treated, the age and relative health of the subject, the route and form of administration, the judgement of the attending medical or veterinary practitioner, and other factors.
The term “pharmaceutical composition” denotes a mixture or solution comprising a therapeutically effective amount of an active pharmaceutical ingredient together with pharmaceutically acceptable excipients to be administered to a mammal, e.g., a human in need thereof.
The present invention relates to (i) a compound of formula (I),
A further embodiment of present invention is (ii) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to (i), wherein
A further embodiment of present invention is (iii) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to (i) or (ii), wherein
A further embodiment of present invention is (iv) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (iii), wherein
A further embodiment of present invention is (v) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (iii), wherein
A further embodiment of present invention is (vii) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (vi), wherein ring B is selected from triazolyl, oxazolyl, oxadiazolyl, and pyrimidinyl.
A further embodiment of present invention is (viii) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (vii), wherein
A further embodiment of present invention is (ix) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (viii), wherein
A further embodiment of present invention is (x) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (viii), wherein R4 is absent or selected from methyl, difluoromethyl, aminomethyl, hydroxymethyl, and carbamoyl.
A further embodiment of present invention is (xi) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (x), wherein R4 is absent or selected from methyl and hydroxymethyl.
A further embodiment of present invention is (xii) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (xi), wherein substituted ring C is
A further embodiment of present invention is (xiii) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (xii), wherein
A further embodiment of present invention is (xiv) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (xiii), wherein
A further embodiment of present invention is (xv) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to any one of (i) or (ii), wherein
A further embodiment of present invention is (xvi) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to (xv), wherein
A further embodiment of present invention is (xvii) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to (xv), wherein
A further embodiment of present invention is (xviii) the compound of formula (I), or a pharmaceutically acceptable salt thereof, according to (xvii), wherein
A further embodiment of present invention is (xix) a compound selected from:
A further embodiment of present invention is (xx) a process for the preparation of a compound having the structure of formula (Ia),
A further embodiment of present invention is (xxi) a process for the preparation of a compound having the structure of formula (Ib),
A further embodiment of present invention is (xxii) a compound of formula (I), formula (Ia), formula (Ib), or a pharmaceutically acceptable salt thereof, when manufactured according to the process of (xx) or (xxi).
A further embodiment of present invention is (xxiii) a pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt thereof according to any one of (i) to (xix), and a pharmaceutically acceptable excipient.
The invention also provides pharmaceutical compositions or medicaments containing the compounds of the invention and a therapeutically inert carrier, diluent or excipient, as well as methods of using the compounds of the invention to prepare such compositions and medicaments. In one example, compounds of formula (I) may be formulated by mixing at ambient temperature at the appropriate pH, and at the desired degree of purity, with physiologically acceptable carriers, i.e., carriers that are non-toxic to recipients at the dosages and concentrations employed into a galenical administration form. The pH of the formulation depends mainly on the particular use and the concentration of compound, but preferably ranges anywhere from about 3 to about 8. In one example, a compound of formula (I) is formulated in an acetate buffer, at pH 5. In another embodiment, the compounds of formula (I) are sterile. The compound may be stored, for example, as a solid or amorphous composition, as a lyophilized formulation or as an aqueous solution.
Compositions are formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners. The “effective amount” of the compound to be administered will be governed by such considerations, and is the minimum amount necessary to activate STING mediated production of Type I IRF responses and proinflammatory cytokines. For example, such amount may be below the amount that is toxic to normal cells, or the mammal as a whole.
In one example, the pharmaceutically effective amount of the compound of the invention administered parenterally per dose will be in the range of about 0.01 to 1000 (e.g., 0.01-100) mg/kg, alternatively about 0.01 to 1000 (e.g., 0.1 to 20) mg/kg of patient body weight per day, with the typical initial range of compound used being 0.3 to 15 mg/kg/day. In another embodiment, oral unit dosage forms, such as tablets and capsules, preferably contain from about 1 to about 1000 (e.g., 25-100) mg of the compound of the invention.
The compounds of the invention may be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural and intranasal, and, if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.
The compounds of the present invention may be administered in any convenient administrative form, e.g., tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain components conventional in pharmaceutical preparations, e.g., diluents, carriers, pH modifiers, sweeteners, bulking agents, and further active agents.
A typical formulation is prepared by mixing a compound of the present invention and a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, e.g., Ansel, Howard C., et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R., et al. Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. The formulations may also include one or more buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present invention or pharmaceutical composition thereof) or aid in the manufacturing of the pharmaceutical product (i.e., medicament).
An example of a suitable oral dosage form is a tablet containing about 1 to 1000 mg (e.g., 25 mg, 50 mg, 100 mg, 250 mg, or 500 mg) of the compound of the invention compounded with about 1 to 1000 (e.g., 90-30) mg anhydrous lactose, about 1 to 1000 (e.g., 5-40) mg sodium croscarmellose, about 1 to 1000 (e.g., 5-30 mg) mg polyvinylpyrrolidone (PVP) K30, and about 1 to 1000 (e.g., 1-10 mg) mg magnesium stearate. The powdered ingredients are first mixed together and then mixed with a solution of the PVP. The resulting composition can be dried, granulated, mixed with the magnesium stearate and compressed to tablet form using conventional equipment. An example of an aerosol formulation can be prepared by dissolving the compound, for example 1 to 500 mg (e.g., 5-400 mg), of the invention in a suitable buffer solution, e.g. a phosphate buffer, adding a tonicifier, e.g. a salt such sodium chloride, if desired. The solution may be filtered, e.g., using a 0.2 micron filter, to remove impurities and contaminants.
An embodiment, therefore, includes a pharmaceutical composition comprising a compound of formula (I), or a stereoisomer or pharmaceutically acceptable salt thereof. In a further embodiment includes a pharmaceutical composition comprising a compound of formula (I), or a stereoisomer or pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier or excipient.
Another embodiment includes a pharmaceutical composition comprising a compound of formula (I) for use in the treatment of cancer.
The following embodiments illustrate typical compositions of the present invention, but serve merely as representative thereof.
A compound of the present invention can be used in a manner known per se as the active ingredient for the production of tablets of the following composition:
A compound of the present invention can be used in a manner known per se as the active ingredient for the production of capsules of the following composition:
The compounds of the invention bind to STING, initiate signaling cascades that induce type I interferon and other pro-inflammatory cytokines and/or co-stimulatory factors. Accordingly, the compounds of the invention are useful for enhancing cancer antigen presentation, contributing to the priming, trafficking and infiltration of T cells, promoting killing of cancers in particular cancer cells. Compounds of the invention are useful for boosting innate immune response in myeloid cells that express STING. Alternatively, compounds of the invention are useful for invigorate T cells in tumors which are T cell present but suppressed, for example by upregulation of co-stimulatory molecules and production of pro-inflammatory cytokines. More broadly, the compounds can be used for the treatment of cancer types which are non-inflamed, e.g. immune desert or immune excluded by enhancing antigen presentation, T cell priming, recruitment/infiltration, and tumor killing.
Another embodiment includes a method of treating cancer in a mammal in need of such treatment, wherein the method comprises administering to said mammal a therapeutically effective amount of a compound of formula (I), a stereoisomer, tautomer, prodrug or pharmaceutically acceptable salt thereof. Cancer includes but is not limited to sarcomas, adenocarcinomas, blastomas, and carcinomas, of the various organ systems, such as those affecting pancreas, liver, lung, breast, lymphoid, stomach, buliarintestinal (e.g. colon), genitourinary tract (e.g. renal urothelial cells), prostate and pharynx.
A further embodiment of present invention is (xxiv) a compound of the invention for use as therapeutically active substance.
A further embodiment of present invention is (xxiv) a compound of the invention for use in the treatment or cancer.
A further embodiment of present invention is (xxvi) the use of a compound of the invention for the treatment of cancer.
A further embodiment of present invention is (xxvii) the use of a compound of the invention for the activation of STING.
A further embodiment of present invention is (xxviii) the use of a compound of the invention for the preparation of a medicament for the treatment of cancer.
A further embodiment of present invention is (xxix) the use of a compound of the invention for the preparation of a medicament for the activation of STING.
A further embodiment of present invention is (xxx) a method for the treatment of cancer, which method comprises administering an effective amount of a compound of the invention.
A further embodiment of present invention is (xxxi) the use of according to (xxvi) or (xxviii), or the method according to (xxx), wherein the cancer is a cancer in pancreas, liver, lung, breast, lymphoid, stomach, buliarintestinal, genitourinary tract, prostate, or pharynx.
The compounds of the present invention can be prepared by any conventional means. Suitable processes for synthesizing these compounds as well as their starting materials are provided in the schemes below and in the examples. All substituents, in particular, ring A, X1 to X3, and R1 to R7 are as defined above unless otherwise indicated. Furthermore, and unless explicitly otherwise stated, all reactions, reaction conditions, abbreviations and symbols have the meanings well known to a person of ordinary skill in organic chemistry.
General synthetic routes for preparing the compound of the invention are shown in scheme 1.
As depicted in Scheme 1, a compound having the structure of formula (Ia) can be prepared via a process comprising one of the following steps:
Also as depicted in Scheme 1, a compound having the structure of formula (Tb) can be prepared via a process comprising the following step:
The invention will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of the invention.
Abbreviations used herein are as follows:
Intermediates and final compounds were purified by flash chromatography using one of the following instruments: i) Biotage SP1 system and the Quad 12/25 Cartridge module. ii) ISCO combi-flash chromatography instrument. Silica gel brand and pore size: i) KP-SIL 60 Å, particle size: 40-60 μm; ii) CAS registry NO: Silica Gel: 63231-67-4, particle size: 47-60 micron silica gel; iii) ZCX from Qingdao Haiyang Chemical Co., Ltd, pore: 200-300 or 300-400.
Intermediates and final compounds were purified by preparative HPLC on reversed phase column using XBridge™ Prep-C18 (5 μm, OBD™ 30×100 mm) column, SunFire™ Prep-C18 (5 μm, OBD™ 30×100 mm) column, Phenomenex Synergi-C18 (10 μm, 25×150 mm) or Phenomenex Gemini-C18 (10 μm, 25×150 mm). Waters AutoP purification System (Sample Manager 2767, Pump 2525, Detector: Micromass ZQ and UV 2487, solvent system: acetonitrile and 0.1% ammonium hydroxide in water; acetonitrile and 0.1% FA in water or acetonitrile and 0.1% TFA in water). Or Gilson-281 purification System (Pump 322, Detector: UV 156, solvent system: acetonitrile and 0.05% ammonium hydroxide in water; acetonitrile and 0.225% FA in water; acetonitrile and 0.05% HCl in water; acetonitrile and 0.075% TFA in water; or acetonitrile and water).
LC/MS spectra of compounds were obtained using a LC/MS (Waters™ Alliance 2795-Micromass ZQ, Shimadzu Alliance 2020-Micromass ZQ or Agilent Alliance 6110-Micromass ZQ), LC/MS conditions were as follows (running time 3 or 1.5 mins):
Mass spectra (MS): generally only ions which indicate the parent mass are reported, and unless otherwise stated the mass ion quoted is the positive mass ion (MH)+.
NMR Spectra were obtained using Bruker Avance 400 MHz, 500 MHz.
The microwave assisted reactions were carried out in a Biotage Initiator Sixty microwave synthesizer. All reactions involving air-sensitive reagents were performed under an argon or nitrogen atmosphere. Reagents were used as received from commercial suppliers without further purification unless otherwise noted.
The title compound was prepared according to the following scheme:
To a solution 2-(aminomethyl)-5-methylpyrazine (A1-a, 7.0 g, 56.84 mmol) in THF (250 mL) was added TEA (12.63 g, 125.04 mmol), the mixture was stirred at r.t. for 1 h, then ethyl 2-chloro-2-oxo-acetate (0.22 g, 1.62 mmol) was added and the mixture was then stirred at r.t. for 2 hrs. The reaction mixture was then poured into water (100 mL) and extracted with DCM (200 mL×4). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated to give compound A1-b (10 g). LCMS (M+H)+: 224.
To a solution of P2O5 (31.79 g, 224 mmol) in POCl3 (100 mL) was added ethyl 2-[(5-methylpyrazin-2-yl) methylamino]-2-oxo-acetate (A1-b, 10 g, 44.8 mmol), the mixture was stirred at 110° C. for 12 hrs. The mixture was then concentrated, the residue was quenched with sat. NaHCO3 solution, extracted with EA (300 mL×3). The combined organic phase was washed with sat. NaHCO3 solution and brine, dried over anhydrous Na2SO4, and concentrated to give compound A1-c (8 g), LCMS (M+H)+: 206.
To a solution of ethyl 6-methylimidazo[1,5-a]pyrazine-3-carboxylate (A1-c, 3.0 g, 14.62 mmol) in DMF (30 mL) was added NBS (2.6 g, 14.62 mmol), the mixture was stirred at 20° C. for 3 hrs. The mixture was then poured into water (80 mL) and extracted with EA (100 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by column chromatography to give compound A1-d (2.9 g). LCMS (M+H)+: 284.
To a solution of ethyl 1-bromo-6-methyl-imidazo[1,5-a]pyrazine-3-carboxylate (A1-d, 2.5 g, 8.8 mmol) in THF (30 mL) was added NaOH (1.41 g, 35.2 mmol) and water (6 mL), the mixture was stirred at 20° C. for 12 hrs. The mixture was then adjusted to pH˜3 with HCl (12 N), the mixture was filtered, the filtered cake was dried to give compound A1-e (2.3 g). LCMS (M+H)+: 256.
To a solution of 1-bromo-6-methyl-imidazo[1,5-a]pyrazine-3-carboxylic acid (A1-e, 2.2 g, 8.59 mmol) in DMF (25 mL) was added DIPEA (3.33 g, 25.78 mmol), HATU (3.26 g, 8.59 mmol), and 2,4-dimethoxybenzylamine (1.55 mL, 10.31 mmol), the mixture was stirred at 20° C. for 3 hrs. The mixture was then poured into water (200 mL), and filtered, the filtered cake was dried to give Intermediate A1 (2.2 g). LCMS (M+H)+: 405.
The title compound was prepared according to the following scheme:
To a solution of diisopropylamine (43.89 mL, 311.34 mmol) in THF (200 mL) was added n-BuLi (2.5 M in hexane solution, 116.75 mL, 291.89 mmol) dropwise at −78° C., the mixture was stirred at −78° C. for 1 h, then a solution of ethyl 1-methylpyrazole-4-carboxylate (A2-a, 30.0 g, 194.59 mmol) in THF (50 mL) was added dropwise at −78° C., the mixture was stirred for 1 h. Then the solution of DMF (33.07 mL, 428.1 mmol) in THF (30 mL) was added dropwise, the mixture was stirred at −78° C. for 2 hrs. The mixture was poured into HCl (1 N, 400 mL), extracted with EA (400 mL×3). The combined organic phase was washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by column chromatography to give compound A2-b (23.5 g). LCMS (M+H)+: 183.
To a solution of ethyl 5-formyl-1-methyl-pyrazole-4-carboxylate (A2-b, 30.0 g, 164.67 mmol) and 2-acetamidoacetic acid (28.92 g, 247.01 mmol) in acetic anhydride (60 mL) was added KOAc (24.21 g, 247.01 mmol), the mixture was stirred at 100° C. for 2 hrs. The mixture was then quenched by sat. NaHCO3 solution (500 mL) and extracted with EA (200 mL×3). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated to give compound A2-c (32.0 g). LCMS (M+H)+: 264.
To a suspension of ethyl 1-methyl-5-[(Z)-(2-methyl-5-oxo-oxazol-4-ylidene) methyl]pyrazole-4-carboxylate (A2-c, 32.0 g, 121.56 mmol) in MeOH (200 mL) was added K2CO3 (33.6 g, 243.11 mmol), the suspension was stirred at 70° C. for 12 hrs. The mixture was diluted with MTBE (500 mL) and filtered, the filtered cake was washed with MTBE, and then suspended in H2O (200 mL). Con. HCl was added to adjust to pH˜1. The mixture was then filtered, the filtered cake was dried to give compound A2-d (20.0 g). LCMS (M+H)+: 194.
SOCl2 (112.74 mL, 1.55 mol) was added to MeOH (400 mL) at 0° C., then 1-methyl-4-oxo-5H-pyrazolo[4,3-c]pyridine-6-carboxylic acid (A2-d, 20.0 g, 103.54 mmol) was added, the mixture was stirred at 60° C. for 16 hrs and then concentrated to give compound A2-e (18.9 g). LCMS (M+H)+: 208.
To a suspension of methyl 1-methyl-4-oxo-5H-pyrazolo[4,3-c]pyridine-6-carboxylate (A2-e, 18.9 g, 91.22 mmol) in ACN (200 mL) was added Py (22.07 mL, 273.66 mmol) and Tf2O (46.07 mL, 273.66 mmol) dropwise at 0° C., the mixture was stirred at 20° C. for 1 h and then used for next step, the compound A2-f (30.9 g) was obtained. LCMS (M+H)+: 340.
To a solution of methyl 1-methyl-4-(trifluoromethylsulfonyloxy) pyrazolo [4,3-c]pyridine-6-carboxylate (A2-f, 30.9 g, 91.08 mmol) in ACN (200 mL) was added LiBr (79.11 g, 910.83 mmol), then TFA (30.44 mL, 409.87 mmol) was added slowly, the mixture was stirred at 20° C. for 16 hrs. The mixture was then quenched with sat. NaHCO3 solution to pH >7, and the mixture was extracted with EA (500 mL×4). The combined organic phase was washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by Prep-HPLC to give compound A2-g (11.5 g). LCMS (M+H)+: 270.
To a suspension of methyl 4-bromo-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxylate (A2-g, 11.5 g, 42.58 mmol) in THF (50 mL) and water (20 mL) was added LiOH·H2O (2.66 g, 63.87 mmol), the mixture was stirred at 20° C. for 1 h. The mixture was then adjusted to pH˜1 with conc. HCl, the mixture was filtered, and the filtered cake was washed with water to give compound A2-h (10.55 g). LCMS (M+H)+: 256.
To a suspension of 4-bromo-1-methyl-pyrazolo [4,3-c]pyridine-6-carboxylic acid (A2-h, 10.55 g, 41.2 mmol) in DMF (40 mL) was added DIPEA (15.97 g, 123.6 mmol), 2, 4-dimethoxybenzylamine (7.43 mL, 49.44 mmol) and HATU (23.5 g, 61.8 mmol), the mixture was stirred at 20° C. for 1 h. The mixture was then diluted with H2O (400 mL) and filtered, the filtered cake was washed with H2O and EA, the solid was concentrated to give Intermediate A2 (15.5 g). LCMS (M+H)+: 405.
The title compound was prepared according to the following scheme:
To a solution of ethylhydrazine dihydrochloride (B1-a, 40.0 g, 303.03 mmol) in DCM (750 mL) was added acetone (24.54 mL, 333.33 mmol) and K2CO3 (105.3 g, 757.58 mmol), the mixture was stirred at 20° C. for 12 hrs. Then the mixture was filtered, the filtered cake was washed with DCM, the filtrate was concentrated to give compound B1-b (25.0 g).
POCl3 (81.68 mL, 873.6 mmol) was added dropwise to DMF (100 mL) at 0° C., the mixture was stirred for 1 h and then cooled to −20° C., then the solution of N-(isopropylideneamino)-ethanamine (B1-b, 35.0 g, 349.44 mmol) in DMF (50 mL) was added dropwise at −20° C. The mixture was stirred at −20° C. for 3 hrs. Then the mixture was further stirred at 80° C. for 2 hrs. The mixture was poured into ice slowly, adjusted to pH-9-10 with 30% aq. NaOH, and then extracted with EA (1.5 L×3). The combined organic phase was dried over anhydrous Na2SO4 and concentrated. The residue was purified by column chromatography to give compound B1-c (26.0 g). LCMS (M+H)+: 139.
To a solution of 1-ethyl-3-methyl-pyrazole-4-carbaldehyde (B1-c, 12.0 g, 86.85 mmol) in DCM (120 mL) was added mCPBA (35.15 g, 173.7 mmol), the mixture was stirred at 40° C. for 1 h. Then the mixture was poured into sat. Na2SO3 solution (800 mL), and extracted with EA (400 mL×3). The combined organic phase was washed with sat. NaHCO3 solution and brine, then dried over anhydrous Na2SO4 and concentrated to give compound B1-d (12.0 g). LCMS (M+H)+: 155.
To a solution of (1-ethyl-3-methyl-pyrazol-4-yl) formate (B1-d, 10.0 g, 64.86 mmol) in MeOH (100 mL) was added TEA (18.13 mL, 129.73 mmol), the mixture was stirred at r.t. for 0.5 h and then concentrated to give compound B1-e (12.0 g). LCMS (M+H)+: 127.
To a solution of 1-ethyl-3-methyl-pyrazol-4-ol (B1-e, 8.0 g, 63.41 mmol) in ACN (100 mL) was added Cs2CO3 (41.34 g, 126.82 mmol) and BnBr (11.31 mL, 95.12 mmol), the mixture was stirred at r.t. for 0.5 h. Then the mixture was filtered, the filterate was poured into water (500 mL) and extracted with EA (800 mL×3). The combined organic phase was washed with sat. NaHCO3 solution and brine, dried over anhydrous Na2SO4 and concentrated, the residue was purified by column chromatography to give compound B1-f (12.0 g). LCMS (M+H)+: 217.
To a solution of 4-benzyloxy-1-ethyl-3-methyl-pyrazole (B1-f, 1.5 g, 6.94 mmol) in THF (160 mL) was added n-BuLi (2.5 M in hexane solution, 4.16 mL, 10.4 mmol) at −78° C. and stirred for 1 h. Then CO2 (3.05 g, 69.35 mmol) was added, the mixture was stirred at r.t. for 1 h and then adjusted to pH˜7 with con. HCl, then concentrated to give compound B1-g (1.8 g). LCMS (M+H)+: 261.
To a solution of 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carboxylic acid (B1-g, 1.8 g, 6.92 mmol) in DCM (20 mL) was added DIPEA (3.63 mL, 20.75 mmol) and isobutyl chloroformate (1.75 mL, 13.83 mmol), the mixture was stirred at r.t. for 2 hrs and then concentrated to give compound B1-h (3.0 g). LCMS (M+H)+: 361.
To a solution of hydrazine (0.59 mL, 18.94 mmol) in THF (5 mL) was added the suspension of isobutoxycarbonyl 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carboxylate (B1-h, 3.0 g, 8.32 mmol) in THF (30 mL) dropwise at 0° C., the mixture was stirred at r.t. for 0.5 h and then concentrated to give compound B1-i (1.6 g). LCMS (M+H)+: 275.
To a solution of 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carbohydrazide (B1-i, 1.6 g, 5.83 mmol) in THF (20 mL) was added DIPEA (1.52 g, 11.67 mmol) and PMBNCS (1.57 g, 8.75 mmol), the mixture was stirred at r.t. for 16 hrs. Then the mixture was diluted with EA (200 mL) and filtered, the filtrate was washed with water, the organic phase was dried with anhydrous Na2SO4 and concentrated to give compound B1-j (3.0 g). LCMS (M+H)+: 454.
To a solution of 1-[(4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carbonyl)amino]-3-[(4-methoxyphenyl)methyl]thiourea (B1-j, 3.0 g, 6.61 mmol) in water (12 mL) was added NaOH (6.0 mL, 18.0 mmol, 3 N), the mixture was stirred at 100° C. for 16 hrs. The mixture was then poured into water (500 mL) and extracted with EA (200 mL×3). The combined organic phase was washed with sat. NaHCO3 solution and brine, dried over anhydrous Na2SO4 and concentrated to give compound B1-k (3.0 g). LCMS (M+H)+: 436.
To a solution of 5-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)-methyl]-1,2,4-triazole-3-thiol (B1-k, 1.5 g, 3.44 mmol) in AcOH (60 mL) was added H2O2 (15.66 g, 138.12 mmol), the mixture was stirred at r.t. for 1 h. The mixture was then poured into ice water (600 mL) and extracted with EA (300 mL×3). The combined organic phase was washed with brine, dried over anhydrous Na2SO4 and concentrated, the residue was purified by column chromatography to give Intermediate B1 (1.02 g). LCMS (M+H)+: 404.
The title compound was prepared according to the following scheme:
To a solution of isobutoxycarbonyl 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carboxylate (B1-h, 1.0 g, 2.77 mmol) in THF (100 mL) was added 1-amino-1-methyl-thiourea (291.8 mg, 2.77 mmol), the mixture was stirred at 50° C. for 12 hrs. Then the mixture was poured into water (100 mL) and extracted with EA (100 mL×3). The combined organic phase was washed with sat. NaHCO3 solution and brine, dried over anhydrous Na2SO4 and concentrated to give compound B2-a (0.85 g). LCMS (M+H)+: 348.
To a solution of 1-[(4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carbonyl)amino]-1-methyl-thiourea (B2-a, 0.8 g, 2.3 mmol) in water (12 mL) was added NaOH (2.09 mL, 6.27 mmol, 3 N), the mixture was stirred at 100° C. for 6 hrs. The mixture was then poured into water (200 mL) and extracted with EA (200 mL×3). The combined organic phase was washed with sat. NaHCO3 solution and brine, dried over anhydrous Na2SO4 and concentrated to give compound B2-b (0.75 g), LCMS (M+H)+: 330.
To a solution of 5-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-2-methyl-1,2,4-triazole-3-thiol (B2-b, 0.7 g, 2.12 mmol) in AcOH (60 mL) was added H2O2 (9.64 g, 85.02 mmol), the mixture was stirred at r.t. for 1 h. The mixture was then poured into ice water (200 mL) and extracted with EA (300 mL×3). The combined organic phase was washed with brine, dried over anhydrous Na2SO4 and concentrated, the residue was purified by column chromatography to give Intermediate B2 (300.0 mg). LCMS (M+H)+: 298.
The title compound was prepared according to the following scheme:
To a solution of ethyl 1-bromo-6-methyl-imidazo[1,5-a]pyrazine-3-carboxylate (A1-d, 338.0 mg, 1.19 mmol) in 1,4-dioxane (1 mL) was added 3-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazole (Intermediate B1, 400.0 mg, 0.99 mmol), Pd(OAc)2 (66.8 mg, 0.3 mmol), CuI (566.4 mg, 2.97 mmol), PCy3HBF4 (145.9 mg, 0.4 mmol) and Cs2CO3 (322.2 mg, 0.99 mmol), the mixture was stirred at 140° C. for 12 hrs. Then the mixture was concentrated, the residue was purified by prep-TLC to give compound 1-a (190.0 mg), LCMS (M+H)+: 607.
To a solution of ethyl 1-[5-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-2-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxylate (1-a, 150.0 mg, 0.25 mmol) in THF (10 mL) was added NaOH (49.5 mg, 1.24 mmol) and water (2 mL), the mixture was stirred at r.t. for 12 hrs. Then the mixture was dissolved in EA (30 mL) and washed with brine, the organic layer was concentrated. The residue was dissolved in DMF (2 mL), then DIEA (67.0 mg, 0.52 mmol), NH4Cl (27.7 mg, 0.52 mmol) and HATU (98.6 mg, 0.26 mmol) were added into, the mixture was stirred at 50° C. for 2 hrs. Then the mixture was concentrated, the residue was purified by reversed-phase column chromatography to give compound 1-b (50.0 mg), LCMS (M+H)+: 578.
The solution of 1-[5-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-2-[(4-methoxyphenyl)-methyl]-1,2,4-triazol-3-yl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (1-b, 50.0 mg, 0.09 mmol) in TFA (1.97 g, 17.31 mmol) was stirred at 80° C. for 0.5 h. Then the mixture was purified by reversed-phase column chromatography to give Example 1 (8.2 mg), LCMS (M+H)+: 368.
1H NMR (400 MHz, DMSO-d6) δ=9.69 (s, 1H), 9.10 (s, 1H), 8.00-7.82 (m, 1H), 4.48 (q, J=6.7 Hz, 2H), 2.58-2.55 (m, 3H), 2.15 (s, 3H), 1.34 (t, J=7.0 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of 3-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-1-methyl-1,2,4-triazole (Intermediate B2, 200.0 mg, 0.67 mmol) and 1-bromo-N-[(2,4-dimethoxyphenyl)methyl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (Intermediate A1, 272.6 mg, 0.67 mmol) in 1,4-dioxane (2 mL) was added Pd(OAc)2 (45.3 mg, 0.2 mmol), CuI (256.2 mg, 1.35 mmol), PCy3HBF4 (99.0 mg, 0.27 mmol) and Cs2CO3 (218.6 mg, 0.67 mmol), the mixture was stirred at 140° C. for 18 hrs. The mixture was then concentrated, the residue was purified by prep-TLC to give compound 2-a (190.0 mg), LCMS (M+H)+: 622.
The solution of 1-[5-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-2-methyl-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (compound 2-a, 0.1 g, 0.16 mmol) in TFA (5.0 mL) was stirred at 80° C. for 2 hrs. The mixture was then concentrated, the residue was dissolved in ACN (5 mL) and stirred at r.t. for 0.5 h. The mixture was filtered, the filtered cake was dried to give Example 2 (34.7 mg). LCMS (M+H)+: 382. 1H NMR (400 MHz, DMSO-d6) δ=9.73 (s, 1H), 9.15 (s, 1H), 8.38 (s, 1H), 7.97 (s, 1H), 7.93 (br s, 1H), 4.45 (s, 3H), 4.45-4.40 (m, 2H), 2.52 (br s, 3H), 2.12 (s, 3H), 1.33 (t, J=7.2 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of methyl 5-formyl-1H-pyrrole-2-carboxylate (3-a, 2.0 g, 13.06 mmol) in ACN (30 mL) was added 1-chloropropan-2-one (1.47 g, 15.93 mmol), Cs2CO3 (5.19 g, 15.93 mmol) and KI (4.33 g, 26.12 mmol), the mixture was stirred at 60° C. for 3 hrs. Then the mixture was quenched with sat. NaHCO3 solution (50 mL), then diluted with water (200 mL) and extracted with EA (250 mL×3). The combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by column chromatography to give compound 3-b (1.5 g).
To a solution of methyl 1-acetonyl-5-formyl-pyrrole-2-carboxylate (3-b, 2.07 g, 9.89 mmol) in EtOH (40 mL) was added NH4OAc (2.288 g, 29.68 mmol), the mixture was stirred at 90° C. for 2 hrs. Then the mixture was concentrated and diluted with water (150 mL), extracted with EA (150 mL×3). The combined organic phase was dried over anhydrous Na2SO4 and concentrated to give compound 3-c (2.1 g), LCMS (M+H)+: 191.
To a solution of methyl 3-methylpyrrolo[1,2-a]pyrazine-6-carboxylate (3-c, 2.1 g, 11.04 mmol) in DMF (20 mL) was added NBS (2.36 g, 13.25 mmol), the mixture was stirred at r.t. for 3 hrs. Then the mixture was diluted with water (100 mL) and extracted with EA (120 mL×3). The combined organic phase was dried over anhydrous Na2SO4 and concentrated. The residue was purified by column chromatography to give compound 3-d (2.02 g). LCMS (M+H)+: 269.
To a solution of methyl 8-bromo-3-methyl-pyrrolo[1,2-a]pyrazine-6-carboxylate (3-d, 1.2 g, 4.46 mmol) in THF (10 mL) was added the solution of NaOH (750.0 mg, 18.75 mmol) in water (3 mL), the mixture was stirred at 40° C. for 12 hrs. Then the mixture was diluted with water (30 mL), HCl (1 N) was added to adjust to pH˜3, and filtered, the filtered cake was dissolved in DMF (4 mL), DIPEA (0.41 mL, 2.35 mmol), HATU (595.9 mg, 1.57 mmol) and 2,4-dimethoxybenzylamine (0.12 mL, 0.78 mmol) were added, the mixture was stirred at r.t. for 2 hrs. Then the mixture was poured into water (20 mL), then filtered, the filtered cake was collected to give compound 3-e (252.0 mg). LCMS (M+H)+: 404.
The solution of K2CO3 (51.2 mg, 0.37 mmol), Pd(OAc)2 (8.3 mg, 0.04 mmol), PivOH (7.6 mg, 0.07 mmol), cataCXium A (13.3 mg, 0.04 mmol), 8-bromo-N-[(2,4-dimethoxyphenyl)-methyl]-3-methyl-pyrrolo[1,2-a]pyrazine-6-carboxamide (3-e, 50.0 mg, 0.12 mmol), 3-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazole (Intermediate B1, 49.9 mg, 0.12 mmol) in toluene (1 mL) was stirred at 120° C. for 12 hrs. Then the mixture was purified by prep-TLC to give compound 3-f (32.0 mg), LCMS (M+H)+: 727.
The solution of 8-[5-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)-methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-3-methyl-pyrrolo[1,2-a]pyrazine-6-carboxamide (3-f, 32.0 mg, 0.04 mmol) in TFA (2.0 mL) was stirred at 70° C. for 2 hrs. Then the mixture was purified by reversed-phase column chromatography to give Example 3 (4.6 mg), LCMS (M+H)+: 367. 1H NMR (400 MHz, DMSO-d6) δ=9.64 (s, 1H), 9.36 (s, 1H), 8.17 (br s, 1H), 4.49 (br d, J=6.8 Hz, 2H), 2.47 (s, 3H), 2.15 (s, 3H), 1.35 (t, J=7.1 Hz, 3H).
The title compound was prepared according to the following scheme:
POCl3 (3.57 mL, 38.18 mmol) was added to the solution of DMF (3.7 mL, 47.73 mmol) in DCM (10 mL) at 0° C., the mixture was stirred at 0° C. for 15 mins, then the solution of ethyl 3-fluoro-1H-pyrrole-2-carboxylate (4-a, 3.0 g, 19.09 mmol) in DCM (5 mL) was added dropwise. The mixture was poured into ice slowly. Then the mixture was adjusted to pH˜9-10 with 30% NaOH aq. and then extracted with EA (50 mL×3). The combined organic phase was dried over anhydrous Na2SO4 and concentrated. The residue was purified by column chromatography to give compound 4-b (1.8 g). (M+H)+: 186.
Compound 4-h was prepared in analogy to the preparation of compound 3-f by using compound 4-b instead of methyl 5-formyl-1H-pyrrole-2-carboxylate (3-a), LCMS (M+H)+: 745.
To a solution of 8-[5-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)-methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-7-fluoro-3-methyl-pyrrolo[1,2-a]-pyrazine-6-carboxamide (4-h, 62.0 mg, 0.08 mmol) in HFIP (2.0 mL) was added MsOH (79.91 mg, 0.83 mmol), the mixture was stirred at r.t. for 1 h. Then the mixture was purified by reversed-phase column chromatography to give Example 4 (22.0 mg). LCMS (M+H)+: 385.
1H NMR (400 MHz, DMSO-d6) δ=9.63 (s, 1H), 9.23 (s, 1H), 7.93-7.71 (m, 1H), 7.57-7.35 (m, 1H), 4.49 (q, J=6.9 Hz, 2H), 2.49 (br s, 3H), 2.16 (s, 3H), 1.36 (t, J=7.1 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of 4-bromo-2-methoxypyridine (5-a, 25.0 g, 132.96 mmol) in DCM (400 mL) was added m-CPBA (32.39 g, 159.56 mmol) in portions, the mixture was stirred at r.t. for 12 hrs. Then the mixture was purified by column chromatography to give compound 5-b (16 g). LCMS (M+H)+: 204.
To a solution of 4-bromo-2-methoxy-1-oxido-pyridin-1-ium (5-b, 16.0 g, 78.42 mmol) in ACN (150 mL) was added TEA (15.84 g, 156.85 mmol) and TMSCN (23.34 g, 235.27 mmol) in portions, the mixture was stirred at 80° C. for 12 hrs under N2. Then the mixture was concentrated, the residue was poured into sat. Na2CO3 solution (200 mL), extracted with EA (400 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by column chromatography to give compound 5-c (12 g), LCMS (M+H)+: 213.
To a solution of 4-bromo-6-methoxy-pyridine-2-carbonitrile (5-c, 3 g, 14.08 mmol) in 1,4-dioxane (30 mL) was added tributyl(1-ethoxyvinyl)tin (5.23 g, 14.48 mmol) and Pd(PPh3)2Cl2 (988.5 mg, 1.41 mmol) under N2. The mixture was stirred at 90° C. for 12 hrs. Then the mixture was quenched with KF solution (45 mL, 1 N). Then HCl solution (60 mL, 1 N) was added to the mixture and stirred at 50° C. for 0.5 h. The mixture was extracted with DCM (100 mL), the organic phase was washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by column chromatography to give compound 5-d (1.8 g). LCMS (M+H)+: 177.
The solution of 4-acetyl-6-methoxy-pyridine-2-carbonitrile (5-d, 1 g, 5.68 mmol) in HCl/MeOH (30 mL, 4 N) was stirred at 80° C. for 12 hrs and then concentrated to give compound 5-e (1.1 g). LCMS (M+H)+: 196.
To a solution of methyl 4-acetyl-6-hydroxy-pyridine-2-carboxylate (5-e, 1.1 g, 5.64 mmol) in ACN (30 mL) was added POBr3 (4.85 g, 16.91 mmol), the mixture was stirred at 80° C. for 2 hrs. Then the mixture was quenched with sat. Na2CO3 solution (50 mL), extracted with EA (50 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated to give compound 5-f (1.2 g). LCMS (M+H)+: 258.
The solution of methyl 4-acetyl-6-bromo-pyridine-2-carboxylate (5-f, 1.2 g, 4.65 mmol) in THF (10 mL) and water (3 mL) was added LiOH·H2O (780.5 mg, 18.6 mmol), the mixture was stirred at r.t. for 2 hrs. Then the mixture was diluted with water (50 mL), HCl solution (1 N) was added to adjust to pH˜5, the mixture was extracted with EA (50 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated to give compound 5-g (900 mg). LCMS (M+H)+: 244.
To a solution of 4-acetyl-6-bromo-pyridine-2-carboxylic acid (5-g, 850.0 mg, 3.48 mmol) in DMF (10 mL) was added DMBNH2 (0.58 mL, 3.83 mmol), DIEA (1.348 g, 10.45 mmol) and HATU (1.588 g, 4.18 mmol), the mixture was stirred at r.t. for 2 hrs. Then the mixture was poured into water (50 mL) and extracted with EA (50 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated. The residue was purified by column chromatography to give compound 5-h (670 mg). LCMS (M+H)+: 393.
To a solution of 4-acetyl-6-bromo-N-[(2,4-dimethoxyphenyl)methyl]pyridine-2-carboxamide (5-h, 150.0 mg, 0.38 mmol) in 1,4-dioxane (3 mL) was added 3-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazole (Intermediate B1, 153.9 mg, 0.38 mmol), Pd(OAc)2 (25.7 mg, 0.11 mmol), CuI (145.3 mg, 0.76 mmol), PCy3HBF4 (56.2 mg, 0.15 mmol) and Cs2CO3 (124.0 mg, 0.38 mmol), the mixture was stirred at 140° C. for 12 hrs. Then the mixture was filtered and concentrated. The residue was purified by prep-TLC to give compound 5-i (40 mg). LCMS (M+H)+: 716.
To a solution of 4-acetyl-6-[5-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]pyridine-2-carboxamide (5-i, 40 mg, 0.06 mmol) in HFIP (1.0 mL) was added MsOH (53.7 mg, 0.56 mmol), the mixture was stirred at r.t. for 1 h. Then the mixture was purified by prep-HPLC to give Example 5 (11.8 mg), LCMS (M+H)+: 356. 1H NMR (400 MHz, DMSO-d6) δ=15.4-15.0 (m, 1H), 8.8-8.7 (m, 1H), 8.65 (d, J=1.2 Hz, 1H), 8.46 (d, J=1.6 Hz, 1H), 8.01 (br s, 1H), 4.41 (q, J=6.8 Hz, 2H), 2.77 (s, 3H), 2.13 (s, 3H), 1.31 (t, J=7.2 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of 4-bromo-6-methoxy-pyridine-2-carbonitrile (5-c, 5.0 g, 23.47 mmol) in DMSO (100 mL) was added imidazole (2.4 g, 35.21 mmol) and K2CO3 (4.87 g, 35.21 mmol), the mixture was stirred at 100° C. for 16 hrs. Then the mixture was poured into H2O (100 mL), and filtered, the filtered cake was dried to give compound 6-a (3.0 g, 14.99 mmol). LCMS (M+H)+: 201.
Example 6 was prepared in analogy to the preparation of Example 5 by using compound 6-a instead of 4-acetyl-6-methoxy-pyridine-2-carbonitrile (compound 5-d). Example 6 (10.7 mg) was obtained. LCMS (M+H)+: 380. 1H NMR (400 MHz, DMSO-d6+D2O) δ=9.70 (br s, 1H), 8.77 (s, 1H), 8.52 (br s, 2H), 7.75 (br s, 1H), 4.43 (q, J=7.1 Hz, 2H), 2.14 (s, 3H), 1.30 (t, J=7.1 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carboxylic acid (B1-g, 5 g, 19.21 mmol) in DMF (50 mL) was added DIEA (7.448 g, 57.63 mmol), NH4Cl (3.08 g, 57.63 mmol) and HATU (10.96 g, 28.81 mmol), the mixture was stirred at r.t. for 2 hrs. Then the mixture was concentrated. The residue was purified by column chromatography to give compound 7-a (2.5 g). LCMS (M+H)+: 260.
To a solution of 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carboxamide (7-a, 2.5 g, 9.64 mmol) in DCM (50 mL) was added TEA (2.95 g, 28.92 mmol), the mixture was cooled to −5° C. Then the solution of TFAA (2.72 mL, 19.28 mmol) in DCM (10 mL) was added into dropwise. Then the mixture was stirred at r.t. for 1 h, then quenched by sat. NaHCO3 solution (800 mL). The mixture was extracted with EA (450 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated. The residue was purified by column chromatography to give compound 7-b (1.5 g). LCMS (M+H)+: 242.
To a suspension of 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carbonitrile (compound 7-b, 1.0 g, 4.14 mmol) in THF (10 mL) was added LiHMDS (1 M in THF solution, 12.43 mL, 12.43 mmol), the mixture was stirred at 30° C. for 48 hrs. Then HCl solution (3 N) was added to adjust to pH˜3, the mixture was stirred at 0° C. for 1 h. Then the mixture was washed with EA. The aqueous phase was adjusted to pH-14 with NaOH solution (3 N), and extracted with EA (900 mL×3). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated to give compound 7-c (1.2 g). LCMS (M+H)+: 259.
To a solution of 4-bromo-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]-pyridine-6-carboxamide (Intermediate A2, 200 mg, 0.49 mmol) in DMF (5 mL) was added TEA (149.8 mg, 1.48 mmol), Pd(PPh3)4 (57.0 mg, 0.05 mmol), tert-butyldimethyl(2-propynyloxy)silane (252.2 mg, 1.48 mmol) and CuI (18.8 mg, 0.1 mmol), the mixture was stirred at 80° C. for 16 hrs under N2. Then the mixture was poured into water (300 mL), extracted with EA (100 mL×3). The combined organic phase was washed with sat. NaHCO3 solution and brine, then dried over anhydrous Na2SO4, and concentrated. The residue was purified by column chromatography to give compound 7-d (200 mg). LCMS (M+H)+: 495.
To a solution of 4-[3-[tert-butyl(dimethyl)silyl]oxyprop-1-ynyl]-N-[(2,4-dimethoxyphenyl)-methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (7-d, 50.0 mg, 0.1 mmol) in MeOH (5 mL) was added NH4F (0.6 g, 16.17 mmol), the mixture was stirred at 60° C. for 0.5 h under N2. Then the mixture was filtered, and concentrated to give compound 7-e (50 mg). LCMS (M+H)+: 381.
To a solution of N-[(2,4-dimethoxyphenyl)methyl]-4-(3-hydroxyprop-1-ynyl)-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (7-e, 30 mg, 0.08 mmol) in DCM (2 mL) was added DMP (50.2 mg, 0.12 mmol), the mixture was stirred for at r.t for 0.5 h under N2. Then the mixture was concentrated. The residue was purified by prep-TLC to give compound 7-f (28 mg).
To a solution of 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carboxamidine (7-c, 23.8 mg, 0.09 mmol) in DCM (20 mL) was added N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-4-(3-oxoprop-1-ynyl)pyrazolo[4,3-c]pyridine-6-carboxamide (7-f, 28 mg, 0.08 mmol), PPh3AuCl3 (39 mg, 0.1 mmol) and K2CO3 (21.2 mg, 0.15 mmol), the mixture was stirred for at r.t. for 1 h. Then the mixture was concentrated. The residue was purified by prep-TLC to give compound 7-g (45 mg). LCMS (M+H)+: 619.
To a solution of 4-[2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)pyrimidin-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (7-g, 40 mg, 0.06 mmol) in HFIP (2.0 mL) was added MsOH (62.1 mg, 0.65 mmol), the mixture was stirred at r.t. for 1 h. Then the mixture was diluted with DMF (1 mL) and purified by reversed-phase column chromatography to give Example 7 (11.2 mg). LCMS (M+H)+: 379. 1H NMR (400 MHz, DMSO-d6) δ=9.34 (br s, 1H), 9.17-9.08 (m, 2H), 8.79 (d, J=5.4 Hz, 1H), 8.59 (br s, 1H), 8.54 (s, 1H), 7.87 (br s, 1H), 4.65 (q, J=7.2 Hz, 2H), 4.23 (s, 3H), 2.19 (s, 3H), 1.31 (t, J=7.2 Hz, 3H).
The title compound was prepared according to the following scheme:
Example 8 was prepared in analogy to the preparation of Example 7 by using Intermediate A1 instead of 4-bromo-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (Intermediate A2). Example 8 (13.5 mg) was obtained. LCMS (M+H+): 379. 1H NMR (400 MHz, DMSO-d6+D2O) δ=9.85 (d, J=1.6 Hz, 1H), 9.10 (s, 1H), 8.89 (d, J=5.4 Hz, 1H), 8.03 (d, J=5.4 Hz, 1H), 4.64 (q, J=7.2 Hz, 2H), 2.48 (br s, 3H), 2.15 (s, 3H), 1.34 (t, J=7.2 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of ethyl 1-bromo-6-methyl-imidazo[1,5-a]pyrazine-3-carboxylate (A1-d, 600 mg, 2.11 mmol) and tributyl(1-ethoxyvinyl)tin (762.7 mg, 2.11 mmol) in DMF (8 mL) was added Pd(PPh3)2Cl2·CH2Cl2 adduct (148.2 mg, 0.21 mmol), the mixture was stirred at 90° C. for 12 hrs under N2. Then the mixture was quenched with KF solution (15 mL, 1 N), and filtered. The filtrate was extracted with DCM (100 mL), the organic phase was washed with brine, dried over anhydrous Na2SO4 and concentrated to give compound 9a (400 mg). LCMS (M+H)+: 276.
To a solution of ethyl 1-(1-ethoxyvinyl)-6-methylimidazo[1,5-a]pyrazine-3-carboxylate (9-a, 400 mg, 1.45 mmol) in THF (8 mL) and water (3 mL) was added NBS (258.6 mg, 1.45 mmol) at 0° C., the mixture was stirred at r.t. for 1 h. Then the mixture was diluted with DCM (50 mL), poured into water (30 mL), and extracted with DCM (30 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated, the residue was purified by column chromatography to give compound 9-b (250 mg). LCMS (M+H)+: 326.
To a solution of ethyl 1-(2-bromoacetyl)-6-methylimidazo[1,5-a]pyrazine-3-carboxylate (9-b, 580 mg, 1.78 mmol) in THF (10 mL) and water (3 mL) was added NaOH (355.7 mg, 8.89 mmol), the mixture was stirred at r.t. for 4 hrs. Then the mixture was concentrated, the residue was dissolved in DMF (5 mL), then DMBNH2 (0.28 mL, 1.87 mmol), DIEA (689.5 mg, 5.33 mmol) and HATU (627.6 mg, 2.67 mmol) were added. The mixture was stirred at r.t. for 12 hrs. Then the mixture was diluted with DCM (100 mL), poured into water (60 mL), and extracted with DCM (100 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated, the residue was purified by column chromatography to give compound 9-c (350 mg). LCMS (M+H)+: 447.
To a solution of 1-(2-bromoacetyl)-N-[(2,4-dimethoxyphenyl)methyl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (9-c, 300 mg, 0.67 mmol) in EA (15 mL) was added silver trifluoromethanesulfonate (258.5 mg, 1.01 mmol) and 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carboxamide (7-a, 260.9 mg, 1.01 mmol), the mixture was stirred at 70° C. for 12 hrs in the dark under N2. Then the mixture was concentrated, the residue was purified by reversed-phase column chromatography to give compound 9-d (100 mg), LCMS (M+H)+: 608.
The solution of 1-[2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (9-d, 98 mg, 0.16 mmol) in TFA (2 mL) was stirred at 80° C. for 4 hrs. The mixture was purified by reversed-phase column chromatography and prep-TLC to give Example 9 (3.6 mg). LCMS (M+H)+: 368. 1H NMR (400 MHz, DMSO-d6) δ=9.64 (s, 1H), 9.05 (s, 1H), 8.79 (s, 1H), 8.63 (s, 1H), 8.05 (br d, J=1.5 Hz, 1H), 7.86 (s, 1H), 4.55-4.49 (m, 2H), 2.46 (br s, 3H), 2.15 (s, 3H), 1.39 (t, J=7.0 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of 1-ethyl-3-methyl-pyrazol-4-ol (B1-e, 25.0 g, 198.16 mmol) in DMF (300 mL) was added K2CO3 (54.7 g, 396.32 mmol) and PMBBr (59.76 g, 297.24 mmol), the mixture was stirred at r.t. for 16 hrs. Then the mixture was poured into water (300 mL) and extracted with EA (300 mL×3). The combined organic phase was washed with sat. NaHCO3 solution and brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by flash chromatography to give compound 10-a (10 g). LCMS (M+H)+: 247.
To a solution of 1-ethyl-4-[(4-methoxyphenyl)methoxy]-3-methyl-pyrazole (10-a, 1.0 g, 4.06 mmol) in THF (10 mL) was added NBS (794.9 mg, 4.47 mmol), the mixture was stirred at r.t. for 3 hrs. Then the mixture was concentrated, the residue was purified by flash column chromatography to give compound 10-b (1.11 g). LCMS (M+H)+: 325.
To a solution of ethyl oxazole-5-carboxylate (10-c, 20 g, 141.72 mmol) in THF (200 mL) and DMF (200 mL) was added LiHMDS (184.24 mL, 184.24 mmol) at −70° C. After stirring for 2 hrs, Br2 (29.44 g, 184.24 mmol) was added dropwise. The mixture was stirred at −70° C. for 2 hrs. Then the mixture was poured into sat. NH4Cl solution (600 mL), extracted with EA (500 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated. The residue was purified by flash chromatography to give Compound 10-d (10 g), LCMS (M+H)+: 220.
To a solution of ethyl 4-bromooxazole-5-carboxylate (10-d, 2.99 g, 13.57 mmol), 4-bromo-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (Intermediate A2, 5.0 g, 12.34 mmol) in toluene (100 mL) and water (25 mL) was added K3PO4 (7.86 g, 37.01 mmol), B2Pin2 (6.27 g, 24.68 mmol), and Pd(dppf)Cl2 (1.81 g, 2.47 mmol), the mixture was stirred at 80° C. for 5 hrs under N2. Then the mixture was concentrated, the residue was purified by flash chromatography to give Compound 10-e (860 mg). LCMS (M+H)+: 466.
To a solution of ethyl 4-[6-[(2,4-dimethoxyphenyl)methylcarbamoyl]-1-methyl-pyrazolo-[4,3-c]pyridin-4-yl]oxazole-5-carboxylate (10-e, 850 mg, 1.83 mmol), 5-bromo-1-ethyl-4-[(4-methoxyphenyl)methoxy]-3-methyl-pyrazole (10-b, 593.9 mg, 1.83 mmol) in toluene (20 mL) was added K2CO3 (756.0 mg, 5.48 mmol), cataCXium A (327.3 mg, 0.91 mmol), PivOH (93.1 mg, 0.91 mmol) and Pd(OAc)2 (123.0 mg, 0.55 mmol), the mixture was stirred at 120° C. for 18 hrs under N2. Then the mixture was filtered, the filtered cake was washed with DCM/MeOH (V/V=20/1). The residue was purified by column chromatography to give Compound 10-f (800 mg), LCMS (M+H)+: 710.
To a solution of ethyl 4-[6-[(2,4-dimethoxyphenyl)methylcarbamoyl]-1-methyl-pyrazolo[4,3-c]pyridin-4-yl]-2-[2-ethyl-4-[(4-methoxyphenyl)methoxy]-5-methyl-pyrazol-3-yl]-oxazole-5-carboxylate (10-f, 800 mg, 1.13 mmol) in THF (20 mL) and TFE (2.0 mL) was added LiBH4 (2.82 mL, 11.27 mmol) at 0° C., the mixture was stirred at 15° C. for 4 hrs under N2. Then the mixture was poured into sat. NH4Cl solution (20 mL), extracted with DCM (50 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated. The residue was purified by flash chromatography to give Compound 10-g (550 mg), LCMS (M+H)+: 668.
To a solution of N-[(2,4-dimethoxyphenyl)methyl]-4-[2-[2-ethyl-4-[(4-methoxyphenyl)-methoxy]-5-methyl-pyrazol-3-yl]-5-(hydroxymethyl)oxazol-4-yl]-1-methyl-pyrazolo[4,3-c]-pyridine-6-carboxamide (10-g, 500 mg, 0.75 mmol) in DCM (10 mL) was added DMP (635.2 mg, 1.5 mmol), the mixture was stirred at r.t. for 1 h under N2. Then the mixture was filtered, the filtered cake was washed with DCM/MeOH (V/V=20/1), the filtrate was concentrated. The residue was purified by column chromatography to give Compound 10-h (250 mg), LCMS (M+H)+: 666.
To a solution of DAST (3.0 mL, 29.42 mmol) in DCM (3 mL) was added the suspension of N-[(2,4-dimethoxyphenyl)methyl]-4-[2-[2-ethyl-4-[(4-methoxyphenyl)methoxy]-5-methyl-pyrazol-3-yl]-5-formyl-oxazol-4-yl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (10-h, 70 mg, 0.11 mmol) in DCM (6 mL) dropwise at 0° C., the mixture was stirred at 0° C. for 8 hrs. Then the mixture was poured into sat. NaHCO3 solution (20 mL), extracted with DCM (20 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated. The residue was purified by prep-TLC to give Compound 10-i (20 mg), LCMS (M+H)+: 688.
To a solution of 4-[5-(difluoromethyl)-2-[2-ethyl-4-[(4-methoxyphenyl)methoxy]-5-methyl-pyrazol-3-yl]oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)-methyl]-1-methyl-pyrazolo [4,3-c]pyridine-6-carboxamide (10-i, 20.0 mg, 0.03 mmol) in HFIP (0.5 mL) was added MsOH (27.9 mg, 0.29 mmol), the mixture was stirred at r.t. for 0.5 h. Then the mixture was diluted with MeOH (1 mL), and purified by reversed-phase column chromatography to give Example 10 (6.5 mg). LCMS (M+H)+: 418. 1H NMR (400 MHz, DMSO-d6) δ=9.07 (s, 1H), 8.79 (s, 1H), 8.43 (s, 1H), 7.87-8.26 (m, 2H), 4.56 (q, J=7.05 Hz, 2H), 4.21 (s, 3H), 2.18 (s, 3H), 1.42 (t, J=7.09 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of isobutoxycarbonyl 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carboxylate (1-h, 7.0 g, 19.42 mmol) in THF (70 mL) was added propargylamine (1.65 g, 29.92 mmol) dropwise at 0° C., the mixture was stirred at r.t. for 12 hrs. Then the mixture was concentrated, the residue was purified by column chromatography to give Compound 11-a (2.4 g). LCMS (M+H)+: 298.
To a solution of 4-benzyloxy-2-ethyl-5-methyl-N-prop-2-ynyl-pyrazole-3-carboxamide (11-a, 2.4 g, 8.07 mmol) in DCM (35 mL) was added NIS (2.72 g, 12.11 mmol), the mixture was stirred at r.t. for 4 hrs. Then the mixture was concentrated to give Compound 11-b (3.0 g), LCMS (M+H)+: 424.
The solution of (5E)-2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-5-(iodomethylene)-4H-oxazole (11-b, 2.8 g, 6.62 mmol) in DCE (15 mL) was stirred at 80° C. for 12 hrs under O2. Then EA (100 mL) was added, the mixture was washed with sat. Na2SO3 solution and brine. The organic phase was dried over anhydrous Na2SO4 and concentrated. The residue was purified by column chromatography to give Compound 11-c (1.0 g), LCMS (M+H)+: 312.
To a solution of 2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazole-5-carbaldehyde (11-c, 900 mg, 2.89 mmol) in EtOH (2 mL) was added NH2OH·HCl (301.3 mg, 4.34 mmol) and KOAc (474.1 mg, 5.78 mmol), the mixture was stirred at r.t. for 3 hrs. The mixture was concentrated (<30° C.), the residue was dissolved in water (30 mL), then extracted with DCM (60 mL×2). The combined organic phase was concentrated to give Compound 11-d (800 mg), LCMS (M+H)+: 327.
To a solution of (5E)-2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazole-5-carbaldehyde oxime (11-d, 800 mg, 2.45 mmol) in AcOH (20 mL) was added Zn (2.745 g, 49.03 mmol) in portions. The mixture was stirred at r.t. for 16 hrs under N2. EA (400 mL) was added to the mixture, then sat. Na2CO3 solution was added to adjust to pH >7. The mixture was extracted with EA (300 mL×2). The combined organic phase was concentrated to give Compound 11-e (600 mg). LCMS (M+H)+: 313.
To a solution of [2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazol-5-yl]methanamine (11-e, 600 mg, 1.92 mmol) in THF (15 mL) was added Boc2O (628 mg, 2.88 mmol) and TEA (582 mg, 5.76 mmol), the mixture was stirred at r.t. for 16 hrs. Then the mixture was poured into water (50 mL) and extracted with EA (80 mL×3). The combined organic phase was washed with sat. NaHCO3 solution and brine, dried over anhydrous Na2SO4 and concentrated, the residue was purified by column chromatography to give Compound 11-f (660 mg), LCMS (M+H)+: 413.
The solution of tert-butyl N-[[2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazol-5-yl]methyl]carbamate (11-f, 560 mg, 1.36 mmol), K2CO3 (562.1 mg, 4.07 mmol), Pd(OAc)2 (91.4 mg, 0.41 mmol), PivOH (83.1 mg, 0.81 mmol), cataCXium A (146.0 mg, 0.41 mmol), 1-bromo-N-[(2,4-dimethoxyphenyl)methyl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (Intermediate A1, 550.2 mg, 1.36 mmol) in toluene (25 mL) was stirred at 115° C. for 12 hrs. DCM (100 mL) was added, then the mixture was filtered, the filtrate was concentrated, the residue was purified by column chromatography and then prep-TLC to give Compound 11-g (27 mg), LCMS (M+H)+: 737.
The solution of tert-butyl N-[[2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[3-[(2,4-dimethoxyphenyl)methylcarbamoyl]-6-methyl-imidazo[1,5-a]pyrazin-1-yl]oxazol-5-yl]methyl]-carbamate (Compound 11-g, 27 mg, 0.04 mmol) in TFA (2.09 g, 18.32 mmol) was stirred at 70° C. for 2 hrs. The mixture was purified by pre-HPLC to give Example 11 (4.7 mg), LCMS (M+H)+: 397. 1H NMR (400 MHz, DMSO-d6+D2O) δ=9.59 (d, J=1.1 Hz, 1H), 8.99 (s, 1H), 4.72 (s, 2H), 4.48 (q, J=6.8 Hz, 2H), 2.43 (s, 3H), 2.13 (s, 3H), 1.36 (t, J=7.2 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of 2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazole-5-carbaldehyde (11-c, 3.5 g, 11.24 mmol) in toluene (150 mL) was added O-diphenylphosphorylhydroxylamine (3.93 g, 16.86 mmol), the mixture was stirred at 100° C. for 2 hrs. Then the mixture was concentrated. The residue was purified by prep-HPLC to give Compound 12-a (1.2 g), LCMS (M+H)+: 309.
The solution of 4-bromo-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (Intermediate A2, 262.9 mg, 0.65 mmol), 2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazole-5-carbonitrile (12-a, 200 mg, 0.65 mmol), Pd(OAc)2 (43.7 mg, 0.19 mmol), CuI (247.1 mg, 1.3 mmol), PCy3·HBF4 (95.5 mg, 0.26 mmol), Cs2CO3 (2.108 g, 0.65 mmol) in 1,4-dioxane (4 mL) was stirred at 140° C. for 18 hrs. Then the mixture was concentrated. The residue was purified by prep-HPLC to give Compound 12-b (70 mg). LCMS (M+H)+: 633.
The solution of 4-[2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-5-cyano-oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (12-b, 10 mg, 0.02 mmol) in TFA (2.0 mL) was stirred at 80° C. for 0.5 h. Then the mixture was concentrated, the residue was purified by prep-HPLC to give Example 12 (1.6 mg), LCMS (M+H)+: 411. 1H NMR (400 MHz, DMSO-d6) δ=10.12 (br s, 1H), 8.78 (s, 1H), 8.40 (s, 1H), 8.39-8.38 (m, 1H), 8.37 (br s, 1H), 8.14 (br s, 1H), 7.83 (br s, 1H), 4.55 (br d, J=6.8 Hz, 2H), 4.20 (s, 3H), 2.11 (s, 3H), 1.38 (t, J=7.1 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of cyclopentanone (13-a, 10.52 mL, 118.88 mmol) in EtOH (60 mL) was added EtONa (42.37 g, 130.77 mmol) dropwise at 0° C., the mixture was stirred at 0° C. for 0.5 h, then diethyl oxalate (16.15 mL, 118.88 mmol) was added dropwise, then the mixture was stirred at r.t. for 12 hrs. The mixture was concentrated (<40° C.), the residue was poured into water (200 mL), then HCl (30 mL, 1 N) was added to adjust to pH˜7, and extracted with EA (500 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated to give Compound 13-b (20 g), LCMS (M+H)+: 185.
To a solution of ethyl 2-(2-hydroxycyclopenten-1-yl)-2-oxo-acetate (13-b, 15.0 g, 81.44 mmol) in EtOH (150 mL) was added ethylhydrazine dihydrochloride (10.83 g, 81.44 mmol) and KOAc (15.98 g, 162.88 mmol), the mixture was stirred at 80° C. for 12 hrs. Then the mixture was concentrated, the residue was extracted with EA (200 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated to give Compound 13-c (8 g), LCMS (M+H)+: 181.
To a solution of 2-ethyl-5,6-dihydro-4H-cyclopenta[c]pyrazole-3-carboxylic acid (13-c, 8.0 g, 44.4 mmol) in THF (80 mL) was added iodoethane (7.1 mL, 88.79 mmol) and K2CO3 (18.38 g, 133.19 mmol), the mixture was stirred at 60° C. for 2 hrs. Water (200 mL) was added, then extracted with EA (300 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated, the residue was purified by column chromatography to give Compound 13-d (5.2 g). LCMS (M+H)+: 209.
Compound 13-h was prepared in analogy to the preparation of Intermediate B1 by using compound 13-d instead of isobutoxycarbonyl 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carboxylate (B1-h). Compound 13-h (400 mg) was obtained. LCMS (M+H)+: 324.
To a solution of 2-ethyl-3-[4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-5,6-dihydro-4H-cyclopenta[c]pyrazole (13-h, 50.0 mg, 0.15 mmol), 4-bromo-N-[(2,4-dimethoxyphenyl)-methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (Intermediate A2, 62.7 mg, 0.15 mmol) in Toluene (5 mL) was added K2CO3 (64.0 mg, 0.46 mmol), cataCXium A (27.7 mg, 0.08 mmol), PivOH (7.9 mg, 0.08 mmol) and Pd(OAc)2 (10.4 mg, 0.05 mmol). The mixture was stirred at 120° C. for 18 hrs under N2. Then the mixture was filtered, the filtered cake was washed by DCM/MeOH (V/V=20/1), the filtrate was combined and concentrated. The residue was purified by prep-TLC to give Compound 13-i (60 mg), LCMS (M+H)+: 648.
To a solution of N-[(2,4-dimethoxyphenyl)methyl]-4-[5-(2-ethyl-5,6-dihydro-4H-cyclopenta[c]pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (Compound 13-i, 45 mg, 0.07 mmol) in HFIP (0.5 mL) was added MsOH (66.7 mg, 0.69 mmol), the mixture was stirred at r.t. for 0.5 h. Then the mixture was diluted with ACN (1 mL), then purified by prep-HPLC to give Example 13 (13.7 mg). LCMS (M+H)+: 378. 1H NMR (400 MHz, DMSO-d6) δ=15.34 (br s, 1H), 8.81 (br s, 1H), 8.75 (d, J=0.8 Hz, 1H), 8.46 (d, J=0.6 Hz, 1H), 7.89 (br s, 1H), 4.66 (q, J=7.1 Hz, 2H), 4.22 (s, 3H), 2.90 (t, J=7.1 Hz, 2H), 2.71-2.63 (m, 2H), 2.47-2.40 (m, 2H), 1.42 (t, J=7.1 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of ethyl 2,4-dioxopentanoate (14-a, 10 g, 63.23 mmol) in EtOH (40 mL) was added hydrazine hydrate (3.48 g, 69.55 mmol) at 0° C., the mixture was stirred at 0° C. for 1 h and then concentrated at r.t. to give compound 14-b (9.6 g). LCMS (M+H)+: 155.
To a solution of ethyl 3-methyl-1H-pyrazole-5-carboxylate (14-b, 9.6 g, 62.27 mmol) in DMF (50 mL) was added (3-bromopropoxy)-tert-butyldimethylsilane (31.54 g, 124.54 mmol), NaI (18.67 g, 124.54 mmol) and K2CO3 (17.21 g, 124.54 mmol), the mixture was stirred at 80° C. for 36 hrs. Then the mixture was diluted with water (800 mL), and extracted with EA (200 mL×3). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated, the residue was purified by flash column chromatography to give compound 14-c (8.0 g), LCMS (M+H)+: 327.
Compound 14-g was prepared in analogy to the preparation of Intermediate B1 by using compound 14-c instead of isobutoxycarbonyl 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carboxylate (B1-h). Compound 14-g (1.6 g) was obtained. LCMS (M+H)+: 328.
To a solution of 3-[5-[4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-3-methyl-pyrazol-1-yl]propan-1-ol (14-g, 800 mg, 2.44 mmol) in DMF (3 mL) was added imidazole (998.2 mg, 14.66 mmol) and TBSCl (1.105 g, 7.33 mmol), the mixture was stirred at 60° C. for 1 h. Then the mixture was concentrated, the residue was purified via flash column chromatography to give compound 14-h (600 mg). LCMS (M+H)+: 442.
To a solution of tert-butyl-[3-[5-[4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-3-methyl-pyrazol-1-yl]propoxy]-dimethyl-silane (14-h, 183.5 mg, 0.45 mmol) in 1,4-dioxane (5 mL) was added 1-bromo-N-[(2,4-dimethoxyphenyl)methyl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (Intermediate A1, 200 mg, 0.45 mmol), Pd(OAc)2 (30.5 mg, 0.14 mmol), CuI (172.5 mg, 0.91 mmol), PCy3·HBF4 (66.7 mg, 0.18 mmol) and Cs2CO3 (147.2 mg, 0.45 mmol), the mixture was stirred at 140° C. for 16 hrs. Then the mixture was dissolved in DCM/MeOH (30 mL/5 mL), filtered, then the filtrate was concentrated, the residue was purified by prep-TLC to give compound 14-i (42.0 mg), LCMS (M+H)+: 766.
The mixture of 1-[5-[2-[3-[tert-butyl(dimethyl)silyl]oxypropyl]-5-methyl-pyrazol-3-yl]-4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)-methyl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (14-i, 40 mg, 0.05 mmol) in TFA (2.5 mL) was stirred at 70° C. for 1.5 hrs. Then the mixture was concentrated, the residue was added into MeOH (1 mL) and NH3·H2O (0.5 mL), then the suspension was stirred at r.t. for 15 mins. The mixture was concentrated, the residue was purified by prep-HPLC to give Example 14 (9.2 mg), LCMS (M+H)+: 382. 1H NMR (400 MHz, DMSO-d6) δ=9.72 (d, J=1.34 Hz, 1H), 9.08 (s, 1H), 8.01-7.82 (m, 2H), 6.62 (s, 1H), 4.67 (t, J=7.03 Hz, 2H), 3.45 (t, J=6.11 Hz, 2H), 2.52 (br s, 3H), 2.22 (s, 3H), 1.98 (br t, J=6.72 Hz, 2H).
The title compound was prepared according to the following scheme:
To a solution of ethylhydrazine dihydrochloride (1.0 g, 7.52 mmol) in EtOH (12 mL) was added K2CO3 (2.28 g, 16.54 mmol) and ethyl 2,4-dioxopentanoate (14-a, 1.31 g, 8.27 mmol) dropwise at 0° C., the mixture was stirred at r.t. for 12 hrs. Then the mixture was concentrated, the residue was extracted with EA (80 mL). The organic layer was washed with water, dried over anhydrous Na2SO4, filtered, and concentrated to give compound 15-a (400 mg). LCMS (M+H)+:183.
To a solution of ethyl 2-ethyl-5-methyl-pyrazole-3-carboxylate (15-a, 2.4 g, 13.17 mmol) in EtOH (30 mL) was added hydrazine hydrate (4.76 g, 80.87 mmol), the mixture was stirred at 80° C. for 16 hrs and then concentrated to give compound 15-b (2 g), LCMS (M+H)+: 169.
The suspension of 2-ethyl-5-methyl-pyrazole-3-carbohydrazide (15-b, 2.0 g, 11.89 mmol) in DMF-DMA (20 mL) was stirred at 100° C. for 1 h. Then the mixture was concentrated. The residue was dissolved in toluene (60 mL), then 4-methoxylbenzylamine (2.456 g, 17.92 mmol) and AcOH (10 mL) were added, the mixture was stirred at 100° C. for 16 hrs. Then the mixture was concentrated, the residue was purified by column chromatography to give compound 15-c (2.1 g). LCMS (M+H)+: 298.
To a solution of 3-(2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazole (15-c, 200 mg, 0.67 mmol) in 1,4-dioxane (2 mL) was added 1-bromo-N-[(2,4-dimethoxyphenyl)methyl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (Intermediate A1, 272.6 mg, 0.67 mmol), Pd(OAc)2 (45.3 mg, 0.2 mmol), CuI (256.2 mg, 1.35 mmol), PCy3·HBF4 (99.0 mg, 0.27 mmol) and Cs2CO3 (218.6 mg, 0.67 mmol), the mixture was stirred at 140° C. for 18 hrs. Then the mixture was filtered and concentrated, the residue was purified by prep-TLC to give compound 15-d (40 mg). LCMS (M+H)+: 622.
The solution of N-[(2,4-dimethoxyphenyl)methyl]-1-[5-(2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (15-d, 30 mg, 0.05 mmol) in TFA (1.5 mL) was stirred at 80° C. for 1 h. The mixture was purified by prep-HPLC to give Example 15 (2.6 mg), LCMS (M+H)+: 352. 1H NMR (400 MHz, DMSO-d6) δ=15.2-14.4 (m, 1H), 9.68 (s, 1H), 9.10 (s, 1H), 8.2-7.7 (m, 2H), 6.8-6.6 (m, 1H), 4.64 (br d, J=6.4 Hz, 2H), 2.5 (s, 3H), 2.23 (s, 3H), 1.41 (t, J=7.2 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of 3-(2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazole (15-c, 1.6 g, 5.38 mmol) in ACN (50 mL) was added Selectflour (3.8 g, 10.76 mmol), the mixture was stirred at 50° C. for 16 hrs. Then the mixture was concentrated, the residue was purified by column chromatography to give compound 16-a (400 mg). LCMS (M+H)+: 316.
Example 16 was prepared in analogy to the preparation of Example 15 by using compound 16-a instead of 3-(2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazole (15-c). Example 16 (13.2 mg) was obtained. LCMS (M+H)+: 370. 1H NMR (400 MHz, DMSO-d6) δ=9.65 (s, 1H), 9.11 (s, 1H), 7.6-8.2 (m, 2H), 4.53 (q, J=7.2 Hz, 2H), 3.31 (br s, 3H), 2.23 (s, 3H), 1.40 (t, J=7.2 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of methyl 3-methyl-1H-pyrazole-5-carboxylate (17-a, 10.0 g, 71.36 mmol) and Cs2CO3 (46.38 g, 142.71 mmol) in ACN (100 mL) was added iodoethane (8.56 mL, 107.04 mmol). The mixture was stirred at 80° C. for 2 hrs. Then the mixture was poured into water (100 ml) and extracted with EA (100 mL×2). The combined organic phase was washed with water and brine, dried over anhydrous Na2SO4, and concentrated. The residue was purified by flash chromatography to give Compound 17-b (5 g). 1H NMR (400 MHz, DMSO-d6) δ=6.64 (s, 1H), 4.42 (q, J=7.21 Hz, 2H), 3.81 (s, 3H), 2.18 (s, 3H), 1.30 (t, J=7.19 Hz, 3H).
To a solution of methyl 2-ethyl-5-methyl-pyrazole-3-carboxylate (17-b, 5 g, 29.73 mmol) in H2SO4 (23.75 mL, 445.92 mmol) was added HNO3 (15.45 mL, 311.14 mmol) dropwise at 0° C. The mixture was stirred at 60° C. for 18 hrs. Then the mixture was poured into ice water (500 mL) and extracted with EA (300 mL×2). The combined organic phase was washed with sat. NaHCO3 solution and brine, dried over anhydrous Na2SO4, and concentrated. The residue was purified by flash column chromatography to give Compound 17-c (3.5 g), LCMS (M+H)+: 214.
To a solution of methyl 2-ethyl-5-methyl-4-nitro-pyrazole-3-carboxylate (17-c, 2.0 g, 9.38 mmol) in MeOH (40 mL) was added Pd/C (1.0 g, 9.38 mmol), the mixture was stirred at r.t. for 16 hrs under H2 balloon. Then the mixture was filtered, the filtered cake was washed with MeOH, the combined filtrate was concentrated to give Compound 17-d (1.7 g). LCMS (M+H)+: 184.
To a solution of methyl 4-amino-2-ethyl-5-methyl-pyrazole-3-carboxylate (17-d, 1.4 g, 7.64 mmol) and Et3N (1.98 mL, 15.28 mmol) in DCM (30 mL) was added BzCl (0.98 mL, 8.41 mmol) at 0° C., the mixture was stirred at r.t. for 2 hrs. Then the mixture was concentrated. The residue was purified by flash column chromatography to give Compound 17-d (2 g), LCMS (M+H)+: 288.
Compound 17-i was prepared in analogy to the preparation of Intermediate B1 by using compound 17-e instead of isobutoxycarbonyl 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carboxylate (B1-h). Compound 17-i (2 g) was obtained. LCMS (M+H)+: 417.
To a solution of N-[1-ethyl-5-[4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-3-methyl-pyrazol-4-yl]benzamide (17-i, 1.0 g, 2.4 mmol) in n-Butanol (20 mL) was added KOH (4.29 g, 76.53 mmol), the mixture was stirred at 130° C. for 16 hrs. Then the mixture was diluted with water (200 mL), and extracted with EA (200 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated to give Compound 17-j (400 mg). LCMS (M+H)+: 313.
To a solution of 1-ethyl-5-[4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-3-methyl-pyrazol-4-amine (17-j, 400 mg, 1.28 mmol), Et3N (388.0 mg, 3.84 mmol), and DMAP (15.9 mg, 0.13 mmol) in THF (0.3 mL) was added Boc2O (837.5 mg, 3.84 mmol), the mixture was stirred at r.t. for 16 hrs. Then the mixture was concentrated, the residue was purified by flash column chromatography to give Compound 17-k (250 mg). LCMS (M+H)+: 513.
Example 17 was prepared in analogy to the preparation of Example 14 by using compound 17-k instead of tert-butyl-[3-[5-[4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-3-methyl-pyrazol-1-yl]propoxy]-dimethyl-silane (14-h). Example 17 (3.7 mg) was obtained. LCMS (M+H)+: 367. 1H NMR (400 MHz, DMSO-d6+D2O) δ=9.63 (br s, 1H), 9.07 (br d, J=3.4 Hz, 1H), 4.51 (br d, J=5.1 Hz, 2H), 2.52 (d, J=1.9 Hz, 3H), 2.15-2.03 (m, 3H), 1.31 (br t, J=6.8 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of 2,5-dichloropyrazine (5.0 g, 33.56 mmol) in DMF (100 mL) was added Cs2CO3 (10.94 g, 33.56 mmol) and ethyl 2-(benzhydrylideneamino)acetate (18-a, 9.87 g, 36.92 mmol), the mixture was stirred at r.t. for 12 hrs. Then the mixture was diluted with EA (900 mL) and water (1 L), the organic layer was washed with water, then dried over anhydrous Na2SO4, and concentrated. The residue was purified by column chromatography to give compound 18-b (8.0 g). LCMS (M+H)+: 380.
To a solution of ethyl 2-(benzhydrylideneamino)-2-(5-chloropyrazin-2-yl)acetate (18-b, 8.0 g, 19.52 mmol) in toluene (36 mL) was added water (12 mL) and HCl (2.0 mL, 24.0 mmol, 12 N), the mixture was stirred at r.t. for 12 hrs. Then the mixture was extracted with toluene (100 mL×3). The aqueous layer was added into water (12 mL) and HCl (1.78 mL, 21.31 mmol, 12 N), the mixture was stirred at 60° C. for 12 hrs and then concentrated to give compound 18-c (4.0 g). LCMS (M+H)+: 144.
Compound 18-h was prepared in analogy to the preparation of Intermediate A1 by using compound 18-c instead of 2-(aminomethyl)-5-methylpyrazine (A1-a). Compound 18-h (360 mg) was obtained. LCMS (M+H)+: 425.
Compound 18-i was prepared in analogy to the preparation of Compound 3-f by using compound 18-h instead of 8-bromo-N-[(2,4-dimethoxyphenyl)methyl]-3-methyl-pyrrolo[1,2-a]pyrazine-6-carboxamide (3-e). Compound 18-i (60 mg) was obtained. LCMS (M+H)+: 748.
Example 18 was prepared in analogy to the preparation of Example 4 by using compound 18-i instead of 8-[5-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-7-fluoro-3-methyl-pyrrolo[1,2-a]pyrazine-6-carboxamide (4-h). Example 18 (2.2 mg) was obtained. LCMS (M+H)+: 388. 1H NMR (400 MHz, DMSO-d6) δ=9.60 (s, 1H), 9.31 (s, 1H), 4.49 (br d, J=7.4 Hz, 2H), 2.15 (s, 3H), 1.34 (t, J=7.2 Hz, 3H).
The title compound was prepared according to the following scheme:
Compound 19-g was prepared in analogy to the preparation of Compound 2-a by using (5-methyl-2-pyridyl)methanamine (19-a) and 3-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazole (Intermediate B1) instead of 2-(aminomethyl)-5-methylpyrazine (A1-a) and 3-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-1-methyl-1,2,4-triazole (Intermediate B2). Compound 19-g (120 mg) was obtained. LCMS (M+H)+: 727.
To a solution of 1-[5-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)-methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-6-methyl-imidazo[1,5-a]pyridine-3-carboxamide (19-g, 120 mg, 0.17 mmol) in HFIP (2.0 mL) was added MsOH (158.5 mg, 1.65 mmol), the mixture was stirred at r.t. for 12 hrs and then concentrated and purified by prep-HPLC to give Example 19 (12.7 mg), LCMS (M+H)+: 367. 1H NMR (400 MHz, DMSO-d6) δ=9.31 (s, 1H), 8.28 (d, J=9.3 Hz, 1H), 7.88-7.45 (m, 2H), 7.25 (br d, J=8.6 Hz, 1H), 4.46 (q, J=6.8 Hz, 2H), 2.36 (s, 3H), 2.14 (s, 3H), 1.33 (t, J=7.1 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of ethyl 2-(6-chloropyrimidin-4-yl)acetate (20-a, 4.0 g, 19.94 mmol) in 1,4-dioxane (100 mL) was added Pd(PPh3)4 (2.304 g, 1.99 mmol), then trimethylaluminum (29.91 mL, 59.81 mmol) was added slowly. The mixture was stirred at 15° C. for 2 hrs. Then the mixture was poured into HCl (100 mL, 1 N), extracted with EA (50 mL). The aqueous phase was adjusted to pH˜9 with sat. K2CO3 solution, extracted with EA (100 mL×3). The combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated to give compound 20-b (3.0 g). LCMS (M+H)+: 181.
To a solution of ethyl 2-(6-methylpyrimidin-4-yl)acetate (20-b, 3.0 g, 16.65 mmol) in acetone (100 mL) was added chloroacetaldehyde (1.634 g, 83.24 mmol) and NaHCO3 (5.594 g, 66.59 mmol), the mixture was stirred at 65° C. for 12 hrs under N2. Then the mixture was concentrated. The residue was purified by column chromatography to give compound 20-c (2.7 g). LCMS (M+H)+: 205.
To a solution of ethyl 3-methylpyrrolo[1,2-c]pyrimidine-5-carboxylate (20-c, 2.7 g, 13.22 mmol) in DCM (50 mL) was added NBS (2.353 g, 13.22 mmol) at 0° C., the mixture was stirred at 15° C. for 1 h. Then the mixture was quenched with sat. NaHCO3 solution (500 mL), extracted with DCM (500 mL×2). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated. The residue was purified by column chromatography to give compound 20-d (1.4 g). LCMS (M+H)+: 283.
To a solution of ethyl 7-bromo-3-methyl-pyrrolo[1,2-c]pyrimidine-5-carboxylate (20-d, 400 mg, 1.41 mmol) in EtOH (6 mL) and water (1 mL) was added NaOH (282.6 mg, 7.06 mmol), the mixture was stirred at 30° C. for 12 hrs. Then water (150 mL) was added, and washed with EA. The aqueous phase was adjusted to pH˜5 with HCl (1 N), extracted with EA (100 mL×3), the combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated to give compound 20-e (150 mg). LCMS (M+H)+: 255.
To a suspension of 7-bromo-3-methyl-pyrrolo[1,2-c]pyrimidine-5-carboxylic acid (20-e, 135 mg, 0.53 mmol) in DMF (1 mL) was added DIEA (0.24 mL, 1.59 mmol), DMBNH2 (0.1 mL, 0.64 mmol) and HATU (301.9 mg, 0.79 mmol), the mixture was stirred at r.t. for 1 h. Then the mixture was poured into H2O (100 mL), extracted with EA (150 mL×3). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by column chromatography to give compound 20-f (80 mg). LCMS (M+H)+: 404.
To a solution of 7-bromo-N-[(2,4-dimethoxyphenyl)methyl]-3-methyl-pyrrolo[1,2-c]pyrimidine-5-carboxamide (compound 20-f, 150.3 mg, 0.37 mmol) in DMF (10 mL) was added 3-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazole (Intermediate B1, 100.0 mg, 0.25 mmol), Li2CO3 (55.0 mg, 0.74 mmol) and CuI (4.7 mg, 0.02 mmol), the mixture was stirred at 140° C. for 12 hrs under N2. Then the mixture was concentrated. The residue was purified by prep-TLC to give compound 20-g (15.0 mg). LCMS (M+H)+: 727.
To a solution of 7-[5-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)-methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-3-methyl-pyrrolo[1,2-c]-pyrimidine-5-carboxamide (compound 20-g, 10.0 mg, 0.01 mmol) in HFIP (0.5 mL) was added MsOH (105.7 mg, 1.1 mmol), the mixture was stirred at r.t. for 3 hrs. Then the mixture was purified by reversed-phase column chromatography to give Example 20 (2.5 mg). LCMS (M+H)+: 367. 1H NMR (400 MHz, DMSO-d6) δ=14.32-13.83 (m, 1H), 10.16 (s, 1H), 9.24-8.88 (m, 1H), 8.05 (br d, J=3.8 Hz, 2H), 7.86-7.67 (m, 1H), 7.18-6.98 (m, 1H), 4.48 (q, J=6.8 Hz, 2H), 2.46 (br s, 3H), 2.15 (s, 3H), 1.34 (t, J=7.2 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of 4-benzyl oxy-1-ethyl-3-methyl-pyrazole (B1-f, 1.0 g, 4.62 mmol) in THF (10 mL) was added NBS (0.91 g, 5.09 mmol), the mixture was stirred at r.t. for 2 hrs. Then the mixture was concentrated, the residue was purified by column chromatography to give compound 21-a (1.3 g). LCMS (M+H)+: 295.
To a solution of 4,6-dimethylpyridin-3-amine (21-b, 10.0 g, 81.85 mmol) in AcOH (200 mL) was added the solution of NaNO2 (6.21 g, 90.04 mmol) in water (20 mL), the mixture was stirred at 15° C. for 12 hrs. Then the mixture was concentrated, diluted with water (200 mL), and then extracted with EA (200 mL×3). The combined organic phase was dried with anhydrous Na2SO4 and concentrated. The residue was purified by column chromatography to give compound 21-c (10.0 g). LCMS (M+H)+: 134.
To a solution of 5-methyl-1H-pyrazolo[3,4-c]pyridine (21-c, 10.0 g, 75.1 mmol) in DMF (200 mL) was added NBS (14.70 g, 82.61 mmol), the mixture was stirred at 30° C. for 1 h. Then the mixture was concentrated, diluted with water (200 mL), and then extracted with EA (200 mL×3). The combined organic phase was dried with anhydrous Na2SO4 and concentrated to give compound 21-d (15.9 g). LCMS (M+H)+: 212.
To a solution of 3-bromo-5-methyl-1H-pyrazolo[3,4-c]pyridine (21-d, 10.0 g, 47.16 mmol) in DMF (20 mL) was added Cs2CO3 (30.73 g, 94.32 mmol) and SEMCl (11.79 g, 70.74 mmol), the mixture was stirred at 60° C. for 2 hrs. Then the mixture was poured into H2O (100 mL) and extracted with EA (200 mL×3). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered and concentrated. The residue was purified by column chromatography to give compound 21-e (4.0 g). LCMS (M+H)+: 342.
To a solution of 2-[(3-bromo-5-methyl-pyrazolo[3,4-c]pyridin-1-yl)methoxy]ethyl-trimethyl-silane (21-e, 4.0 g, 11.69 mmol) in DMF (30 mL) and MeOH (30 mL) was added DPPF (1.29 g, 2.34 mmol), Pd(OAc)2 (262.3 mg, 1.17 mmol) and TEA (2.95 g, 29.21 mmol), the mixture was stirred at 60° C. under CO (45 psi) for 16 hrs. Then the mixture was filtered and washed with MeOH. The filtrate was concentrated, the residue was purified by column chromatography to give compound 21-f (3.7 g). LCMS (M+H)+: 322.
To a solution of methyl 5-methyl-1-(2-trimethylsilylethoxymethyl)pyrazolo[3,4-c]pyridine-3-carboxylate (21-f, 3.4 g, 10.58 mmol) in THF (20 mL) was added the solution of LiOH·H2O (887.7 mg, 21.15 mmol) in water (20 mL), the mixture was stirred at r.t. for 1 h. Then the mixture was cooled to 0° C., HCl solution (0.5 N) was added dropwise to adjust to pH˜4. The suspension was filtered, the filtered cake was washed with water and dried to give compound 21-g (3.2 g). LCMS (M+H)+: 308.
To a solution of 5-methyl-1-(2-trimethylsilylethoxymethyl)pyrazolo[3,4-c]pyridine-3-carboxylic acid (21-g, 3.2 g, 10.41 mmol) in DMF (60 mL) was added DIPEA (6.72 g, 52.05 mmol), HATU (5.93 g, 15.61 mmol), and 2,4-dimethoxybenzylamine (2.61 g, 15.61 mmol), the mixture was stirred at r.t. for 1 h. Then the mixture was poured into H2O (30 mL) and extracted with EA (30 mL×3). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered and concentrated. The residue was purified by column chromatography to give compound 21-h (4.0 g). LCMS (M+H)+: 457.
To a solution of N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-1-(2-trimethylsilylethoxymethyl)pyrazolo[3,4-c]pyridine-3-carboxamide (21-h, 4.0 g, 8.76 mmol) in THF (50 mL) was added TBAF (25.0 mL, 25.0 mmol, 1 N in THF), the mixture was stirred at 60° C. for 16 hrs. Then the mixture was concentrated, the residue was dissolved in DCM (100 mL). Then sat. NaHCO3 solution (100 mL) was added, the mixture was extracted with DCM (100 mL×3). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered and the concentrated. The residue was purified by column chromatography to give compound 21-i (2.5 g). LCMS (M+H)+: 327.
To a solution of N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-1H-pyrazolo[3,4-c]pyridine-3-carboxamide (21-i, 500.0 mg, 1.53 mmol) and ethyl 4-bromooxazole-5-carboxylate (10-d, 674.2 mg, 3.06 mmol) in DMF (6 mL) was added CuI (58.4 mg, 0.31 mmol), (1R,2R)-1,2-diaminocyclohexane (35.0 mg, 0.31 mmol) and K3PO4 (650.4 mg, 3.06 mmol), the mixture was stirred at 120° C. for 12 hrs. Then the mixture was filtered and concentrated, the residue was purified by column chromatography to give compound 21-j (200.0 mg). LCMS (M+H)+: 466.
To a solution of ethyl 4-[3-[(2,4-dimethoxyphenyl)methylcarbamoyl]-5-methyl-pyrazolo[3,4-c]pyridin-1-yl]oxazole-5-carboxylate (21-j, 200.0 mg, 0.43 mmol), 4-benzyloxy-5-bromo-1-ethyl-3-methyl-pyrazole (21-a, 200.0 mg, 0.68 mmol) in 1,4-dioxane (1 mL) was added Pd(OAc)2 (45.6 mg, 0.2 mmol), PCy3HBF4 (99.7 mg, 0.27 mmol), CuI (258.1 mg, 1.36 mmol) and Cs2CO3 (220.2 mg, 0.68 mmol), the mixture was stirred at 140° C. for 18 hrs under N2. Then the mixture was filtered, the filtered cake was washed with DCM/MeOH (V/V=20/1), the filtrate was concentrated, the residue was purified by prep-TLC to give compound 21-k (30.0 mg). LCMS (M+H)+: 680.
To a solution of ethyl 2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[3-[(2,4-dimethoxyphenyl)methylcarbamoyl]-5-methyl-pyrazolo[3,4-c]pyridin-1-yl]oxazole-5-carboxylate (21-k, 30.0 mg, 0.04 mmol) in THF/H2O/MeOH (V/V/V=1/1/1, 1 mL) was added NaOH (17.6 mg, 0.44 mmol), the mixture was stirred at 30° C. for 2 hrs. Then HCl solution (1 N) was added to adjust to pH˜4, the mixture was extracted with EA (30 mL×3), the combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered and concentrated to give compound 21-1 (28.0 mg). LCMS (M+H)+: 652.
To a solution of 2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[3-[(2,4-dimethoxyphenyl)methylcarbamoyl]-5-methyl-pyrazolo[3,4-c]pyridin-1-yl]oxazole-5-carboxylic acid (21-1, 28.0 mg, 0.04 mmol) in DMSO (1 mL) was added AcOH (0.5 mg, 0.01 mmol) and Ag2CO3 (2.4 mg, 0.01 mmol), the mixture was stirred at 80° C. for 12 hrs. Then the mixture was filtered, the filtrate was poured into water (10 mL), extracted with EA (10 mL×2). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered and concentrated to give compound 21-m (20.0 mg). LCMS (M+H)+: 608.
To a solution of 1-[2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (21-m, 20.0 mg, 0.03 mmol) in HFIP (2.0 mL) was added MsOH (31.6 mg, 0.33 mmol), the mixture was stirred at 50° C. for 12 hrs. Then the mixture was purified by reversed-phase column chromatography to give Example 21 (3.5 mg). LCMS (M+H)+: 368. 1H NMR (400 MHz, DMSO-d6+D2O) δ=9.57 (s, 1H), 8.69 (s, 1H), 8.06 (s, 1H), 4.49 (q, J=7.2 Hz, 2H), 2.66 (s, 3H), 2.15 (s, 3H), 1.39 (t, J=7.2 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of 4-methoxybenzylchloride (6.35 g, 40.55 mmol), DIEA (8.73 g, 67.59 mmol) and KI (2.8 g, 16.9 mmol) in MeCN (50 mL) was added 3-bromo-1H-1,2,4-triazole (22-a, 5.0 g, 33.79 mmol), the mixture was stirred at 80° C. for 16 hrs. Then the mixture was concentrated, the residue was purified by column chromatography to give compound 22-b (5.0 g). LCMS (M+H)+: 268.
Compound 22-d was prepared in analogy to the preparation of compound 21-k by using compound 22-b instead of ethyl 4-bromooxazole-5-carboxylate (10-d). Compound 22-d (10 mg) was obtained. LCMS (M+H)+: 728.
Example 22 was prepared in analogy to the preparation of Example 21 by using compound 22-d instead of 1-[2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (21-m). Example 22 (2.1 mg) was obtained. LCMS (M+H)+: 368. 1H NMR (400 MHz, DMSO-d6+D2O) δ=9.64 (s, 1H), 8.12 (s, 1H), 4.48 (q, J=7.2 Hz, 2H), 2.69 (s, 3H), 2.17 (s, 3H), 1.36 (t, J=7.2 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of methyl 2-oxopropanoate (16.51 g, 161.71 mmol) in MeOH (250 mL) was added AcONa (13.27 g, 161.71 mmol) and ethyl 2-hydrazinoacetate hydrochloride (23-a, 25.0 g, 161.71 mmol), the mixture was stirred at r.t. for 18 hrs. Then H2O (80 mL) was added to the mixture. Then the mixture was concentrated to remove MeOH. The aqueous phase was extracted with EA (400 mL×3). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered and concentrated to give compound 23-b (28.5 g). LCMS (M+H)+: 203.
To a solution of methyl (2E)-2-[(2-ethoxy-2-oxo-ethyl)hydrazono]propanoate (23-b, 47.0 g, 232.43 mmol) in MeOH (470 mL) was added MeONa (139.46 mL, 697.29 mmol, 5 N in MeOH), the mixture was stirred at 70° C. for 16 hrs. Then HCl (2 N) was added to adjust to pH˜7 at 0° C., then the mixture was extracted with EA (500 mL×3). The combined organic phase was dried with anhydrous Na2SO4, filtered and concentrated to give compound 23-c (17.0 g). LCMS (M+H)+: 157.
Compound 23-d was prepared in analogy to the preparation of compound B1-f by using compound 23-c and Na2CO3 instead of 1-ethyl-3-methyl-pyrazol-4-ol (B1-e) and Cs2CO3. Compound 23-d (16.6 g) was obtained. LCMS (M+H)+: 247.
To a solution of methyl 4-benzyloxy-3-methyl-1H-pyrazole-5-carboxylate (23-d, 20.0 g, 81.21 mmol) in DMF (200 mL) was added K2CO3 (22.45 g, 162.43 mmol), 2-bromoethoxy-tert-butyl-dimethyl-silane (23.31 g, 97.46 mmol) and NaI (10.64 g, 71 mmol), the mixture was stirred at 40° C. for 28 hrs. Then additional 2-bromoethoxy-tert-butyl-dimethyl-silane (9.71 g, 40.61 mmol) and K2CO3 (11.22 g, 81.21 mmol) were added, the mixture was stirred at 40° C. for another 16 hrs. Then the mixture was poured into water (600 mL), extracted with EA (300 mL×3). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered, and concentrated, the residue was purified by column chromatography to give compound 23-e (9.0 g). LCMS (M+H)+: 405.
Compound 23-i was prepared in analogy to the preparation of intermediate B1 by using compound 23-e instead of isobutoxycarbonyl 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carboxylate (B1-h). Compound 23-i (3.6 g) was obtained. LCMS (M+H)+: 420.
To a solution of 2-[4-benzyloxy-5-[4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-3-methyl-pyrazol-1-yl]ethanol (23-i, 4.0 g, 9.54 mmol) in DCM (100 mL) was added imidazole (3.9 g, 57.21 mmol) and TBSCl (4.31 g, 28.61 mmol), the mixture was stirred at 60° C. for 1 h. Then the mixture was concentrated, the residue was purified by column chromatography to give compound 23-j (2.0 g). LCMS (M+H)+: 534.
Compound 23-k was prepared in analogy to the preparation of compound 1-a by using compound 23-j and Intermediate A2 instead of ethyl 1-bromo-6-methyl-imidazo[1,5-a]pyrazine-3-carboxylate (A1-d) and 3-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)-methyl]-1,2,4-triazole (Intermediate B1). Compound 23-k (150 mg) was obtained. LCMS (M+H)+: 858.
To a solution of 4-[5-[4-benzyloxy-2-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-5-methyl-pyrazol-3-yl]-4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)-methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (23-k, 150 mg, 0.17 mmol) in MeOH (4 mL) was added NH4F (232.5 mg, 6.12 mmol), the mixture was stirred at 60° C. for 12 hrs. Then the mixture was concentrated, the residue was purified by column chromatography to give compound 23-1 (120 mg). LCMS (M+H)+: 744.
To a solution of 4-[5-[4-benzyloxy-2-(2-hydroxyethyl)-5-methyl-pyrazol-3-yl]-4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (23-1, 33 mg, 0.04 mmol), CMBP (0.07 mL, 0.22 mmol) in anhydrous toluene (1.5 mL) was added imidazole (15.1 mg, 0.22 mmol), the mixture was stirred at 60° C. for 12 hrs. Then the mixture was concentrated, the residue was purified by column chromatography to give compound 23-m (10 mg). LCMS (M+H)+: 794.
Example 23 was prepared in analogy to the preparation of Example 21 by using compound 23-m instead of 1-[2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (21-m). Example 23 (2.5 mg) was obtained. LCMS (M+H)+: 434. 1H NMR (400 MHz, DMSO-d6) δ=15.36 (s, 1H), 8.91-8.81 (m, 2H), 8.68 (s, 1H), 8.49 (s, 1H), 8.15 (s, 1H), 7.90 (s, 1H), 7.46 (s, 2H), 4.95 (d, J=4.4 Hz, 2H), 4.69 (d, J=4.4 Hz, 2H), 4.24 (s, 3H), 2.11 (s, 3H).
The title compound was prepared according to the following scheme:
To a solution of 4-[5-[4-benzyloxy-2-(2-hydroxyethyl)-5-methyl-pyrazol-3-yl]-4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (23-1, 200 mg, 0.27 mmol) in DCM (5 mL) was added DMP (228.1 mg, 0.54 mmol) at 0° C., the mixture was stirred at 30° C. for 1 h. Then sat. Na2SO3 solution (50 mL) and sat. NaHCO3 solution (100 mL) were added into the mixture at 0° C. The mixture was extracted with DCM (100 mL×3). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered, and concentrated, the residue was purified by prep-TLC to give compound 24-a (100 mg). LCMS (M+H)+: 742.
To a solution of 4-[5-[4-benzyloxy-5-methyl-2-(2-oxoethyl)pyrazol-3-yl]-4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (24-a, 100 mg, 0.13 mmol) in EtOH (3 mL) was added (cyanomethyl)triphenylphosphonium chloride (91.13 mg, 0.27 mmol), then TEA (0.06 mL, 0.4 mmol) in EtOH (3 mL) was added to the mixture dropwise at 0° C. The mixture was stirred at 30° C. for 3 hrs. Then the mixture was purified by prep-TLC to give compound 24-b (80 mg). LCMS (M+H)+: 765.
To a solution of 4-[5-[4-benzyloxy-2-[(E)-3-cyanoallyl]-5-methyl-pyrazol-3-yl]-4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (24-b, 80 mg, 0.1 mmol) in MeOH (5 mL) was added Pd/C (300 mg) under N2, the mixture was stirred at r.t. for 2 hrs under H2 (15 psi). Then the mixture was purified by prep-TLC to give compound 24-c (50 mg). LCMS (M+H)+: 677.
The solution of 4-[5-[2-(3-cyanopropyl)-4-hydroxy-5-methyl-pyrazol-3-yl]-4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (24-c, 12 mg, 0.02 mmol) and TFA (8.1 mg, 0.07 mmol) in DCM (1.5 mL) was stirred at 30° C. for 3 hrs. Then the mixture was purified by reversed-phase column chromatography to give Example 24 (2.2 mg). LCMS (M+H)+: 407. 1H NMR (400 MHz, DMSO-d6) δ=15.38 (s, 1H), 8.88 (s, 1H), 8.84 (s, 1H), 8.48 (s, 1H), 8.10 (s, 1H), 7.88 (s, 1H), 4.53-4.50 (m, 2H), 4.22 (s, 3H), 3.31 (s, 2H), 2.15 (s, 3H), 2.15-2.07 (m, 2H).
The title compound was prepared according to the following scheme:
To a solution of 2-[4-benzyloxy-5-[4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-3-methyl-pyrazol-1-yl]ethanol (23-i, 1.0 g, 2.38 mmol) in THF (10 mL) was added Py (565 mg, 7.15 mmol) and Ms2O (830.5 mg, 4.77 mmol) at 0° C., the mixture was stirred at r.t. for 1 h. Then H2O (20 mL) was added into the mixture, the mixture was extracted with DCM (100 mL×2). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered, and concentrated to give compound 25-a (1.0 g). LCMS (M+H)+: 498.
To a solution of 2-[4-benzyloxy-5-[4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-3-methyl-pyrazol-1-yl]ethyl methanesulfonate (25-a, 800 mg, 1.61 mmol) in DMF (8 mL) was added NaCN (760 mg, 15.51 mmol), the mixture was stirred at 60° C. for 18 hrs. Then sat. NaHCO3 solution (50 mL) was added into the mixture, the mixture was extracted with DCM (50 mL×2). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered, and concentrated, the residue was purified by column chromatography to give compound 25-b (330.0 mg). LCMS (M+H)+: 429.
Compound 25-c was prepared in analogy to the preparation of compound 3-f by using compound 25-b and Intermediate A2 instead of 8-bromo-N-[(2,4-dimethoxyphenyl)-methyl]-3-methyl-pyrrolo[1,2-a]pyrazine-6-carboxamide (3-e) and 3-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazole (Intermediate B1). Compound 25-c (120 mg) was obtained. LCMS (M+H)+: 753.
Example 25 was prepared in analogy to the preparation of Example 24 by using compound 25-c instead of 4-[5-[4-benzyloxy-2-[(E)-3-cyanoallyl]-5-methyl-pyrazol-3-yl]-4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (24-b). Example 25 (12.4 mg) was obtained. LCMS (M+H)+: 393. 1H NMR (400 MHz, DMSO-d6) δ=15.39 (s, 1H), 8.87 (s, 2H), 8.47 (s, 1H), 8.15 (s, 1H), 7.88 (br d, J=1.2 Hz, 1H), 4.75 (br t, J=6.6 Hz, 2H), 4.23 (s, 3H), 3.09 (t, J=6.6 Hz, 2H), 2.16 (s, 3H).
The title compound was prepared according to the following scheme:
To a solution of ethyl 1-bromo-6-methyl-imidazo[1,5-a]pyrazine-3-carboxylate (A1-d, 1.9 g, 6.69 mmol) in CCl4 (40 mL) was added NBS (1.31 g, 7.36 mmol) and AIBN (109.8 mg, 0.67 mmol), the mixture was stirred at 90° C. for 16 hrs. Then the mixture was concentrated, the residue was purified by column chromatography to give compound 26-a (2.1 g). LCMS (M+H)+: 362.
To a solution of ethyl 1-bromo-6-(bromomethyl)imidazo[1,5-a]pyrazine-3-carboxylate (26-a, 2.1 g, 5.78 mmol) in 1,4-dioxane (20 mL) was added CaCO3 (2.89 g, 28.92 mmol) and water (20 mL), the mixture was stirred at 90° C. for 16 hrs. Then water (100 mL) was added into the mixture, the mixture was extracted with EA (50 mL×3). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered and concentrated to give compound 26-b (1.6 g). LCMS (M+H)+: 300.
To a solution of ethyl 1-bromo-6-(hydroxymethyl)imidazo[1,5-a]pyrazine-3-carboxylate (26-b, 1.6 g, 5.33 mmol) in DCM (40 mL) was added DMP (3.61 g, 8.53 mmol), the mixture was stirred at r.t. for 1 h. Then the mixture was filtered, the filtered cake was washed with DCM, the filtrate was concentrated, the residue was purified by column chromatography to give compound 26-c (1.0 g). LCMS (M+H)+: 298.
To a solution of ethyl 1-bromo-6-formyl-imidazo[1,5-a]pyrazine-3-carboxylate (26-c, 1.0 g, 3.35 mmol) in DCM (20 mL) was added DAST (1.62 g, 10.06 mmol) dropwise at 0° C., the mixture was stirred at r.t. for 1 h. Then sat. NaHCO3 solution (100 mL) was added into the mixture, the mixture was extracted with DCM (100 mL×3). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered and concentrated, the residue was purified by column chromatography to give compound 26-d (650 mg). LCMS (M+H)+: 320.
Example 26 was prepared in analogy to the preparation of Example 5 by using compound 26-d instead of methyl 4-acetyl-6-bromo-pyridine-2-carboxylate (5-f). Example 26 (17.1 mg) was obtained. LCMS (M+H)+: 404. 1H NMR (400 MHz, DMSO-d6+D2O) δ=9.78 (s, 1H), 9.53 (s, 1H), 7.15 (t, J=54.4 Hz, 1H), 4.56-4.39 (m, 2H), 2.15 (s, 3H), 1.34 (t, J=7.0 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carboxylic acid (B1-g, 1.0 g, 3.84 mmol) in DMF (10 mL) was added DIEA (1.48 g, 11.53 mmol), NH4Cl (616.5 mg, 11.53 mmol) and HATU (2.19 g, 5.76 mmol), the mixture was stirred at 50° C. for 2 hrs. Then the mixture was concentrated, the residue was purified by reversed-phase column chromatography to give compound 27-a (850 mg). LCMS (M+H)+: 260.
To a solution of 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carboxamide (27-a, 0.6 g, 2.31 mmol) in DCM (5 mL) was added TEA (708.1 mg, 6.94 mmol), then TFAA (972 mg, 4.63 mmol) in DCM (2 mL) was added dropwise at −5° C., the mixture was stirred at 0° C. for 1 h. Then NaHCO3 solution (5%, 100 mL) was added into the mixture, the mixture was extracted with EA (250 mL×2). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered and concentrated, the residue was purified by column chromatography to give compound 27-b (0.5 g). LCMS (M+H)+: 242.
The mixture of 4-benzyloxy-2-ethyl-5-methyl-pyrazole-3-carbonitrile (27-b, 0.4 g, 1.66 mmol), hydroxyamine hydrochloride (0.34 g, 4.97 mmol) and TEA (2.29 mL, 16.58 mmol) in EtOH (10 mL) was stirred at 80° C. for 1 h. Then the mixture was extracted with EA (150 mL×2). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered and concentrated, the residue was purified by column chromatography to give compound 27-c (450 mg). LCMS (M+H)+: 275.
To a solution of 4-benzyloxy-2-ethyl-N-hydroxy-5-methyl-pyrazole-3-carboxamidine (27-c, 0.4 g, 1.46 mmol) in trimethyl orthoformate (154.7 mg, 1.46 mmol) was added p-toluenesulfonic acid monohydrate (0.04 g, 0.21 mmol), the mixture was stirred at 110° C. for 1 h. Then NaHCO3 solution (5%, 100 mL) was added into the mixture, the mixture was extracted with EA (150 mL×2). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered and concentrated, the residue was purified by column chromatography to give compound 27-d (350 mg). LCMS (M+H)+: 285.
To a solution of 3-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-1,2,4-oxadiazole (27-d, 200 mg, 0.7 mmol) in toluene (5 mL) was added 7-bromo-N-[(2,4-dimethoxyphenyl)methyl]-3-methyl-pyrrolo[1,2-c]pyrimidine-5-carboxamide (20-f, 200 mg, 0.49 mmol), Pd(OAc)2 (15.8 mg, 0.07 mmol), PPh3 (36.9 mg, 0.14 mmol) and AgOAc (350.3 mg, 2.11 mmol), the mixture was stirred at 120° C. for 12 hrs. Then the mixture was concentrated, the residue was purified by prep-TLC to give compound 27-e (18 mg). LCMS (M+H)+: 608.
Example 27 was prepared in analogy to the preparation of Example 21 by using compound 27-e instead of 1-[2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (21-m). Example 27 (2.1 mg) was obtained. LCMS (M+H)+: 368. 1H NMR (400 MHz, DMSO-d6+D2O) δ=10.11 (s, 1H), 8.43 (s, 1H), 8.17 (s, 1H), 4.39 (d, J=6.8 Hz, 2H), 2.54 (s, 3H), 2.15 (s, 3H), 1.34 (t, J=7.2 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of 1-ethyl-4-[(4-methoxyphenyl)methoxy]-3-methyl-pyrazole (10-a, 2.0 g, 8.12 mmol) in THF (20 mL) was added n-BuLi (6.5 mL, 16.24 mmol, 2.5 N in hexane) at −78° C. dropwise, the mixture was stirred at −78° C. for 1 h. Then tributyltin chloride (5.7 g, 17.51 mmol) was added into the mixture at −78° C., the mixture was stirred at r.t. for 3 hrs. Then the mixture was poured into water (100 mL), HCl (1 N) was added to adjust to pH-5. The mixture was extracted with DCM (50 mL×3). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered and concentrated, the residue was purified by column chromatography to give compound 28-a (3.7 g). LCMS (M+H)+: 537.
To a solution of ethyl 2-bromooxazole-5-carboxylate (1.0 g, 4.55 mmol) in 1,4-Dioxane (40 mL) was added tributyl-[2-ethyl-4-[(4-methoxyphenyl)methoxy]-5-methyl-pyrazol-3-yl]stannane (28-a, 3.65 g, 6.82 mmol) and Pd(PPh3)2Cl2 (0.32 g, 0.45 mmol), the mixture was stirred at 110° C. for 18 hrs. Then the mixture was filtered and washed with EA. The filtrate was concentrated, the residue was purified by column chromatography to give compound 28-b (720 mg). LCMS (M+H)+: 386.
To a solution of 8-bromo-N-[(2,4-dimethoxyphenyl)methyl]-7-fluoro-3-methyl-pyrrolo[1,2-a]pyrazine-6-carboxamide (4-g, 322 mg, 0.76 mmol) and ethyl 2-(1-ethyl-4-((4-methoxybenzyl)oxy)-3-methyl-1H-pyrazol-5-yl)oxazole-5-carboxylate (28-b, 294.8 mg, 0.76 mmol) in toluene (1 mL) was added KOPiv (321.8 mg, 2.29 mmol), CataCXium A Pd G3 (167.1 mg, 0.23 mmol) and PivOH (46.8 mg, 0.46 mmol), the mixture was stirred at 120° C. for 12 hrs. Then the mixture was diluted with DCM/MeOH (V/V=10/1, 20 mL), filtered and concentrated, the residue was purified by prep-TLC to give compound 28-c (30 mg). LCMS (M+H)+: 727.
Example 28 was prepared in analogy to the preparation of Example 21 by using compound 28-c instead of ethyl 2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)-4-[3-[(2,4-dimethoxyphenyl)methylcarbamoyl]-5-methyl-pyrazolo[3,4-c]pyridin-1-yl]oxazole-5-carboxylate (21-k). Example 28 (3.7 mg) was obtained. LCMS (M+H)+: 385. 1H NMR (400 MHz, DMSO-d6) δ=9.55 (s, 1H), 9.15 (s, 1H), 8.77 (br s, 1H), 8.58 (s, 1H), 7.79 (s, 1H), 7.41 (s, 1H), 4.50 (q, J=6.8 Hz, 2H), 2.45 (s, 3H), 2.15 (s, 3H), 1.37 (t, J=7.2 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of 2,4-dimethyl-5-nitro-pyridine (29-a, 20.0 g, 131.45 mmol) in DMF (400 mL) was added DMF-DMA (32.29 g, 271.0 mmol), the mixture was stirred at 100° C. for 2 hrs. Then the mixture was purified by column chromatography to give compound 29-b (24.0 g). LCMS (M+H)+: 208.
The mixture of (E)-N,N-dimethyl-2-(2-methyl-5-nitro-4-pyridyl)ethenamine (29-b, 24.0 g, 115.81 mmol) and Pd/C (10.0 g) in EtOH (300 mL) was stirred under H2 (15 psi) at r.t. for 12 hrs. Then the mixture was filtered through dicalite and concentrated to give compound 29-c (20.0 g). LCMS (M+H)+: 149.
The mixture of 1-hydroxy-5-methyl-pyrrolo[2,3-c]pyridine (29-c, 20.0 g, 134.99 mmol) and Pd/C (10.0 g) in EtOH (300 mL) was stirred under H2 (15 psi) at r.t. for 12 hrs. Then the mixture was filtered through dicalite and concentrated, the residue was purified by reversed-phase column chromatography to give compound 29-d (10.0 g). LCMS (M+H)+: 133.
To a solution of NBS (4.04 g, 22.7 mmol) in ACN (50 mL) was added 5-methyl-1H-pyrrolo[2,3-c]pyridine (29-d, 3.0 g, 22.7 mmol), the mixture was stirred at r.t. for 12 hrs. Then the mixture was filtered and concentrated to give compound 29-e (3.0 g). LCMS (M+H)+: 211.
To a solution of 3-bromo-5-methyl-1H-pyrrolo[2,3-c]pyridine (29-e, 3.0 g, 14.21 mmol) in MeOH (60 mL) was added Pd(OAc)2 (1.27 g, 5.69 mmol), dicyclohexyl(3-dicyclohexylphosphaniumylpropyl)phosphanium;ditetrafluoroborate (DCPP, 3.48 g, 5.69 mmol) and K2CO3 (5.88 g, 42.64 mmol), the mixture was stirred at 80° C. for 12 hrs under CO (50 Psi). Then the mixture was filtrated and concentrated, the residue was purified by reversed-phase column chromatography to give compound 29-f (2.0 g). LCMS (M+H)+: 191.
To a solution of methyl 5-methyl-1H-pyrrolo[2,3-c]pyridine-3-carboxylate (29-f, 2.0 g, 10.52 mmol) in THF/H2O/MeOH (V/V/V=1/1/1, 30 mL) was added NaOH (4.21 g, 105.15 mmol), the mixture was stirred at 60° C. for 12 hrs. Then the mixture was purified by reversed-phase column chromatography give compound 29-g (1.7 g). LCMS (M−H)−: 175.
Compound 29-h was prepared in analogy to the preparation of compound 21-h by using compound 29-g instead of 5-methyl-1-(2-trimethylsilylethoxymethyl)pyrazolo[3,4-c]pyridine-3-carboxylic acid (21-g). Compound 29-h (1.1 g) was obtained. LCMS (M−H)−: 324.
Example 29 was prepared in analogy to the preparation of Example 21 by using compound 29-h instead of N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-1H-pyrazolo[3,4-c]pyridine-3-carboxamide (21-i). Example 29 (2.9 mg) was obtained. LCMS (M+H)+: 367. 1H NMR (400 MHz, DMSO-d6+D2O) δ 9.29 (s, 1H), 9.03 (s, 1H), 8.79 (s, 1H), 8.39 (s, 1H), 4.44 (q, J=7.0 Hz, 2H), 2.73 (s, 3H), 2.13 (s, 3H), 1.34 (t, J=7.2 Hz, 3H).
The title compound was prepared according to the following scheme:
Compound 30-b was prepared in analogy to the preparation of compound 25-d by using compound 4-g instead of 4-bromo-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (Intermediate A2). Compound 30-b (16 mg) was obtained. LCMS (M+H)+: 680.
Example 30 was prepared in analogy to the preparation of Example 21 by using compound 30-b instead of 1-[2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (21-m). Example 30 (1.5 mg) was obtained. LCMS (M+H)+: 410. 1H NMR (400 MHz, DMSO-d6) δ=14.20-13.95 (m, 1H), 9.61 (s, 1H), 9.38-9.26 (m, 1H), 9.22 (s, 1H), 7.89-7.77 (m, 1H), 7.57-7.39 (m, 1H), 4.77 (br t, J=6.6 Hz, 2H), 3.11 (t, J=6.6 Hz, 2H), 2.48 (s, 3H), 2.18 (s, 3H).
The title compound was prepared according to the following scheme:
To a mixture of NaH (871.9 mg, 21.8 mmol) in THF (15 mL) was added the solution of 3-methyl-1H-pyrazole-4-carbaldehyde (32-a, 2.0 g, 18.16 mmol) in THF (15 mL) dropwise at 0° C., the mixture was stirred at 0° C. for 30 min. Then SEMCl (4.54 g, 27.25 mmol) was added into the mixture dropwise. The mixture was stirred at r.t. for 16 hrs. Then the mixture was poured into water (50 mL), extracted with EA (50 mL×3). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered, and concentrated. The residue was purified by column chromatography to give compound 32-b (3.8 g). LCMS (M+H)+: 241.
Compound 31-e was prepared in analogy to the preparation of compound B1-f by using compound 31-b instead of 1-ethyl-3-methyl-pyrazole-4-carbaldehyde (B1-c). Compound 31-e (3.13 g) was obtained. LCMS (M+H)+: 319.
To a solution of 2-[(4-benzyloxy-3-methyl-pyrazol-1-yl)methoxy]ethyl-trimethyl-silane (31-e, 2.0 g, 6.28 mmol) in DCM (30 mL) was added TFA (14.8 g, 129.8 mmol), the mixture was stirred at r.t. for 12 hrs. Then the mixture was concentrated, the residue was dissolved in DCM (10 mL). Then sat. NaHCO3 solution was added to adjust to pH˜8. The mixture was then extracted with DCM (50 mL×3), the combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered, and concentrated. The residue was purified by column chromatography to give compound 31-f (1.2 g). LCMS (M+H)+: 189.
To a solution of 4-benzyloxy-3-methyl-1H-pyrazole (31-f, 500 mg, 2.66 mmol) in DMF (10 mL) was added 1-iodopropane (903.1 mg, 5.31 mmol) and Cs2CO3 (2.59 g, 7.97 mmol), the mixture was stirred at 80° C. for 16 hrs. Then the mixture was diluted with water (10 mL), extracted with EA (30 mL×3). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered, and concentrated. The residue was purified by prep-TLC to give compound 31-g (450 mg). LCMS (M+H)+: 231.
Compound 31-h was prepared in analogy to the preparation of compound 21-a by using compound 31-g instead of 4-benzyloxy-1-ethyl-3-methyl-pyrazole (B1-f). Compound 31-h (200 mg) was obtained. LCMS (M+H)+: 309.
The solution of N-[(2,4-dimethoxyphenyl)methyl]-1-[1-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (22-c, 120 mg, 0.23 mmol), 4-benzyloxy-5-bromo-3-methyl-1-propyl-pyrazole (31-h, 72.2 mg, 0.23 mmol), Pd(OAc)2 (10.5 mg, 0.05 mmol), CuI (14.3 mg, 0.14 mmol), PPh3 (33.5 mg, 0.09 mmol) and K2CO3 (96.7 mg, 0.7 mmol) in toluene (3 mL) was stirred at 120° C. for 12 hrs under N2. Then the mixture was diluted with DCM (10 mL) and filtered. The filtrate was concentrated, the residue was purified by prep-TLC to give compound 31-i (10 mg). LCMS (M+H)+: 742.
Example 31 was prepared in analogy to the preparation of Example 21 by using compound 31-i instead of 1-[2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (21-m). Example 31 (1.7 mg) was obtained. LCMS (M+H)+: 382. 1H NMR (400 MHz, DMSO-d6) δ=14.20 (br s, 1H), 9.64 (s, 1H), 9.20 (s, 1H), 8.06 (s, 2H), 7.78 (br s, 1H), 4.43 (t, J=6.8 Hz, 2H), 2.67 (s, 3H), 2.18 (s, 3H), 1.86-1.75 (m, 2H), 0.87 (t, J=7.2 Hz, 3H).
Example 32 was prepared in analogy to the preparation of Example 31 by using bromomethylcyclopropane instead of 1-iodopropane. Example 32 (3.4 mg) was obtained. LCMS (M+H)+: 394. 1H NMR (400 MHz, DMSO-d6) δ=9.61 (s, 1H), 8.04-7.95 (m, 2H), 7.70 (br s, 1H), 4.37 (d, J=6.8 Hz, 2H), 2.65 (s, 3H), 2.15 (s, 3H), 1.37-1.28 (m, 1H), 0.47-0.41 (m, 2H), 0.40-0.35 (m, 2H).
The title compound was prepared according to the following scheme:
To a solution of 4-benzyloxy-3-methyl-1H-pyrazole (31-f, 500 mg, 2.66 mmol) in 1,4-Dioxane (20 mL) was added cyclopropylboronic acid (456.4 mg, 5.31 mmol), Cu(OAc)2 (482.5 mg, 2.66 mmol), DMAP (1.29 g, 10.63 mmol) and Py (524.6 mg, 6.64 mmol), the mixture was stirred at 100° C. for 16 hrs under O2. Then the mixture was diluted with water (50 mL), extracted with EA (30 mL×2). The combined organic phase was washed with brine, dried with anhydrous Na2SO4, filtered, and concentrated. The residue was purified by column chromatography to give compound 33-a (540 mg). LCMS (M+H)+: 229.
Example 33 was prepared in analogy to the preparation of Example 31 by using compound 33-a instead of 4-benzyloxy-3-methyl-1-propyl-pyrazole (31-g). Example 33 (1.5 mg) was obtained. LCMS (M+H)+: 380. 1H NMR (400 MHz, DMSO-d6) δ=14.31 (br s, 1H), 9.67 (d, J=1.2 Hz, 1H), 9.15 (br s, 1H), 8.10-8.00 (m, 2H), 7.77 (br s, 1H), 4.20-4.16 (m, 1H), 2.67 (s, 3H), 2.15 (s, 3H), 1.07-1.05 (m, 2H), 0.99-0.96 (m, 2H).
The title compound was prepared according to the following scheme:
To a solution of 4-[5-[4-benzyloxy-5-methyl-2-(2-oxoethyl)pyrazol-3-yl]-4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (24-a, 50 mg, 0.07 mmol) and PPh3 (22.1 mg, 0.08 mmol) in DMF (1 mL) was added sodium 2-chloro-2,2-difluoro-acetate (16.1 mg, 0.11 mmol), the mixture was stirred at 100° C. for 12 hrs. Then the mixture was added water (50 mL), extracted with EA (50 mL×3). The combined organic phase was washed with water, dried with anhydrous Na2SO4, filtered, and concentrated. The residue was purified by prep-TLC to give compound 34-a (12 mg). LCMS (M+H)+: 776.
Example 34 was prepared in analogy to the preparation of Example 21 by using compound 34-a instead of 1-[2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (21-m). Example 34 (1.3 mg) was obtained. LCMS (M+H)+: 416. 1H NMR (400 MHz, METHANOL-d4) δ=8.86 (s, 1H), 8.43 (s, 1H), 5.26 (br d, J=7.6 Hz, 2H), 4.90 (s, 1H), 4.23 (s, 3H), 2.25 (s, 3H).
Example 35 was prepared in analogy to the preparation of Example 23 by using 1,2,3-triazole instead of imidazole. Example 35 (7.1 mg) was obtained. LCMS (M+H)+: 435. 1H NMR (400 MHz, DMSO-d6) δ=15.33 (br s, 1H), 8.90-8.86 (m, 2H), 8.46 (s, 1H), 8.05 (br s, 1H), 7.87 (br s, 1H), 7.66 (s, 2H), 4.98 (t, J=6.2 Hz, 2H), 4.87 (t, J=6.2 Hz, 2H), 4.22 (s, 3H), 2.12 (s, 3H).
Example 36 was prepared in analogy to the preparation of Example 23 by using 1,2,3-triazole instead of imidazole. Example 36 (5.1 mg) was obtained. LCMS (M+H)+: 435. 1H NMR (400 MHz, DMSO-d6) δ=15.32 (br s, 1H), 8.86 (s, 2H), 8.46 (s, 1H), 8.08 (s, 1H), 7.8-7.9 (m, 2H), 7.57 (s, 1H), 4.96 (t, J=5.2 Hz, 2H), 4.86 (t, J=5.2 Hz, 2H), 4.23 (s, 3H), 2.12 (s, 3H).
The title compound was prepared according to the following scheme:
To a solution of 4-[3-[2-(3,3-difluoroallyl)-4-hydroxy-5-methyl-pyrazol-3-yl]-1H-1,2,4-triazol-5-yl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (Example 34, 10 mg, 0.02 mmol) in THF (3 mL) was added Pd/C (10 mg), the mixture was stirred at 30° C. for 12 hrs under H2 balloon. Then the mixture was purified by reversed-phase column chromatography to give Example 37 (2.9 mg). LCMS (M+H)+: 418. 1H NMR (400 MHz, DMSO-d6) δ=15.39 (s, 1H), 8.86 (d, J=1.6 Hz, 1H), 8.83 (s, 1H), 8.47 (s, 1H), 8.07 (s, 1H), 7.87 (d, J=2.6 Hz, 1H), 6.34-6.00 (m, 1H), 4.64 (t, J=7.4 Hz, 2H), 4.22 (s, 3H), 2.53-2.52 (m, 1H), 2.46-2.45 (m, 1H), 2.15 (s, 3H).
The title compound was prepared according to the following scheme:
Compound 38-a was prepared in analogy to the preparation of compound 23-m by using 1,2,4-triazole instead of imidazole. The residue was purified by prep-TLC (DCM/MeOH=30/1), The minor spot (Rf=0.25) was defined as Compound 38-a (10 mg) without further structure conformation. LCMS (M+H)+: 795.
Example 38 was prepared in analogy to the preparation of Example 21 by using compound 38-a instead of 1-[2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (21-m). Example 38 (2.0 mg) was obtained. LCMS (M+H)+: 435. 1H NMR (400 MHz, DMSO-d6) δ=15.21 (s, 1H), 8.72 (s, 1H), 8.24 (s, 1H), 8.09 (s, 1H), 7.93 (s, 1H), 7.86 (s, 1H), 7.61 (s, 1H), 7.34 (s, 1H), 4.87 (t, J=5.8 Hz, 2H), 4.63 (t, J=5.8 Hz, 2H), 2.97 (d, J=4.4 Hz, 3H), 2.09 (s, 3H).
The title compound was prepared according to the following scheme:
Compound 39-a was prepared in analogy to the preparation of compound 23-m by using 1,2,4-triazole instead of imidazole. The residue was purified by prep-TLC (DCM/MeOH=30/1), The major spot (Rf=0.3) was defined as Compound 39-a (20 mg) without further structure conformation. LCMS (M+H)+: 795.
Example 39 was prepared in analogy to the preparation of Example 21 by using compound 39-a instead of 1-[2-(4-benzyloxy-2-ethyl-5-methyl-pyrazol-3-yl)oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (21-m). Example 39 (6.7 mg) was obtained. LCMS (M+H)+: 435. 1H NMR (400 MHz, DMSO-d6) δ=15.33 (s, 1H), 8.90-8.82 (m, 2H), 8.47 (s, 1H), 8.24 (s, 1H), 8.05 (br s, 1H), 7.89-7.85 (m, 1H), 7.83 (s, 1H), 4.91 (t, J=6.2 Hz, 2H), 4.66 (t, J=6.2 Hz, 2H), 4.23 (s, 3H), 2.13 (s, 3H).
The title compound was prepared according to the following scheme:
To a solution of 4-benzyloxy-3-methyl-1H-pyrazole (31-f, 500 mg, 2.66 mmol) in DMF (4 mL) was added Cs2CO3 (2596 mg, 7.97 mmol) and 1-bromo-2-phenylethane (1.09 mL, 7.97 mmol). The mixture was stirred at 100° C. for 12 hrs. The reaction mixture was cooled and diluted with EtOAc (20 mL) and water (20 mL). The organic layer was separated, the aqueous phase was extracted with EtOAc (20 mL×2). The combined organic layer was dried over Na2SO4, and concentrated to give a residue, which was purified via prep-TLC to give compound 40-a (500 mg). LCMS (M+H)+: 293. 1H NMR (400 MHz, CHLOROFORM-d) 6=7.40-7.35 (m, 4H), 7.29-7.22 (m, 4H), 7.13-7.07 (m, 2H), 6.75 (s, 1H), 4.82 (s, 2H), 4.20-4.11 (m, 2H), 3.08 (t, J=7.4 Hz, 2H), 2.22 (s, 3H).
To a solution of compound 40-a (500.0 mg, 1.71 mmol) in THF (5 mL) was added NBS (365 mg, 2.05 mmol). The mixture was stirred at 20° C. for 1 h. The reaction mixture was concentrated and purified by prep-TLC to afford compound 40-b (450.0 mg). LCMS (M+H)+: 371. 1H NMR (400 MHz, CHLOROFORM-d) 6=7.36 (s, 4H), 7.33-7.22 (m, 4H), 7.19-6.99 (m, 2H), 4.90 (s, 2H), 4.24 (dd, J=7.2, 8.4 Hz, 2H), 3.14-3.01 (m, 2H), 2.10 (s, 3H).
A mixture of compound 40-b (200.0 mg, 0.54 mmol), N-[(2,4-dimethoxyphenyl)methyl]-1-[1-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (22-c, 276 mg, 0.54 mmol), Pd(OAc)2 (24 mg, 0.11 mmol), CuI (61 mg, 0.32 mmol), PPh3 (56 mg, 0.22 mmol), K2CO3 (223 mg, 1.62 mmol) in toluene (10 mL) was stirred at 120° C. for 12 hrs under N2 atmosphere. The reaction mixture was diluted with DCM (10 mL) and filtered. The filtrate was concentrated and purified by prep-TLC to afford compound 40-c (10 mg). LCMS (M+H)+: 804.
A solution of compound 40-c (10 mg, 0.01 mmol) and MsOH (12 mg, 0.12 mmol) in HFIP (0.3 mL) was stirred at 30° C. for 3 hrs. The reaction mixture was purified by HPLC to give Example 40 (1.8 mg). LCMS (M+H)+: 444, 1H NMR (400 MHz, DMSO-d6) δ=14.14 (s, 1H), 9.62 (s, 1H), 9.23 (s, 1H), 8.05 (s, 1H), 8.00-7.92 (m, 1H), 7.88-7.79 (m, 1H), 7.28-7.21 (m, 2H), 7.21-7.10 (m, 3H), 4.67 (t, J=7.4 Hz, 2H), 3.09-3.05 (m, 2H), 2.67 (s, 3H), 2.18 (s, 3H).
Example 41 was prepared in analogy to the preparation of Example 40 by using compound 41-c instead of compound 40-b.
The compound 41-c was prepared according to the following scheme:
To a solution of 4-benzyloxy-3-methyl-1H-pyrazole (31-f, 500 mg, 2.66 mmol) in acetonitrile (5 mL) was added Cs2CO3 (2.6 g, 7.97 mmol, 3.0 eq) and 1-bromo-3-fluoropropane (1.12 g, 7.97 mmol). The mixture was stirred at 80° C. for 12 hrs. The mixture was then filtered and concentrated. The residue was purified by prep-TLC to afford compound 41-a (250 mg) and compound 41-b (150 mg).
Compound 41-a, LCMS (M+H)+: 249. 1H NMR (400 MHz, CHLOROFORM-d) 6=7.45-7.31 (m, 5H), 6.97 (s, 1H), 4.89 (s, 2H), 4.44 (t, J=5.6 Hz, 1H), 4.32 (t, J=5.6 Hz, 1H), 4.08 (t, J=6.7 Hz, 2H), 2.25-2.09 (m, 5H).
Compound 41-b, LCMS (M+H)+: 249. 1H NMR (400 MHz, CHLOROFORM-d) 6=7.43-7.29 (m, 5H), 7.24 (s, 1H), 4.94 (s, 2H), 4.44 (t, J=5.6 Hz, 1H), 4.32 (t, J=5.6 Hz, 1H), 4.11 (t, J=6.8 Hz, 2H), 2.29-2.06 (m, 5H).
To a solution of 4-benzyloxy-1-(3-fluoropropyl)-3-methyl-pyrazole (41-a, 220 mg, 0.89 mmol) in THF (2 mL) was added NBS (205 mg, 1.15 mmol). The mixture was stirred at 25° C. for 1 h. The mixture was concentrated, and the residue was purified via flash chromatography on silica gel to afford compound 41-c (220 mg). LCMS (M+H)+: 327.
Example 41 (2.4 mg), LCMS (M+H)+: 400, 1H NMR (400 MHz, DMSO-d6) δ=14.37-14.11 (m, 1H), 9.64 (d, J=1.0 Hz, 1H), 9.44-9.15 (m, 1H), 8.08 (s, 2H), 7.80 (br s, 1H), 4.60 (t, J=7.0 Hz, 2H), 4.54 (t, J=5.8 Hz, 1H), 4.42 (t, J=5.8 Hz, 1H), 2.68 (s, 3H), 2.23-2.13 (m, 5H).
Example 42 was prepared in analogy to the preparation of Example 40 by using 2-bromoethyl methyl ether instead of 1-bromo-2-phenylethane in step 1.
Example 42, LCMS (M+H)+: 398, 1H NMR (400 MHz, DMSO-d6) δ=14.23 (br s, 1H), 9.63 (s, 1H), 9.30-9.08 (m, 1H), 8.08 (s, 2H), 7.79 (s, 1H), 4.64 (t, J=5.6 Hz, 2H), 3.70 (t, J=5.8 Hz, 2H), 3.18 (s, 3H), 2.67 (s, 3H), 2.17 (s, 3H).
Example 43 was prepared in analogy to the preparation of Example 40 by using 1-bromobutane instead of 1-bromo-2-phenylethane in step 1.
Example 43 (1.5 mg), LCMS (M+H)+: 396, 1H NMR (400 MHz, DMSO-d6) δ=14.20 (br s, 1H), 9.64 (br s, 1H), 9.19 (br s, 1H), 8.05 (s, 2H), 7.78 (br s, 1H), 4.47 (br t, J=7.2 Hz, 2H), 2.67 (s, 3H), 2.18 (s, 3H), 1.82-1.72 (m, 2H), 1.32-1.26 (m, 2H), 0.87 (br t, J=7.2 Hz, 3H).
Example 44 was prepared in analogy to the preparation of Example 40 by using 1-bromo-3-methoxypropane instead of 1-bromo-2-phenylethane in step 1.
Example 44 (11.7 mg), LCMS (M+H)+: 412, 1H NMR (400 MHz, DMSO-d6) δ=9.65 (s, 1H), 8.01 (s, 2H), 7.78-7.67 (m, 1H), 7.22-6.84 (m, 1H), 4.53 (t, J=7.0 Hz, 2H), 3.29-3.29 (m, 2H), 3.18 (s, 3H), 2.65 (s, 3H), 2.15 (s, 3H), 2.05-1.96 (m, 2H).
The title compound was prepared according to the following scheme:
To a solution of methyl 4-benzyloxy-3-methyl-1H-pyrazole-5-carboxylate (1.5 g, 6.09 mmol) in toluene (15 mL) was added CMBP (7.36 g, 30.46 mmol) and 3-pentyn-1-ol (2.81 mL, 30.46 mmol), the mixture was then stirred at 80° C. for 12 hrs. The reaction was concentrated to give a residue. The residue was purified via column chromatography to afford compound 45-a (1.8 g). LCMS (M+H)+: 313.
To a solution of compound 45-a (1.8 g, 5.76 mmol) in methanol (20 mL) was added hydrazine hydrate (2.94 g, 57.63 mmol) at r.t. Then the mixture was stirred at 80° C. for 4 hrs. The mixture was concentrated to give compound 45-b (1.8 g). LCMS (M+H)+: 313.
To a yellow solution of compound 45-b (1.7 g, 5.44 mmol) and DIPEA (2.69 mL, 16.33 mmol) in THF (80 mL) was added 4-methoxybenzyl isothiocyanate (1.95 g, 10.88 mmol) at r.t. The mixture was stirred for 12 hrs and concentrated to obtained compound 45-c (2.3 g). LCMS (M+H)+: 492.
A yellow solution of compound 45-c (2.3 g, 4.68 mmol) in 3N NaOH (25 mL) was stirred at 100° C. for 3 h. The mixture was washed with EA (3×900 mL). The combined organic layer was washed with water (3×800 mL) and brine (1050 mL), dried over anhydrous sodium sulfate and concentrated, the residue was purified by column chromatography to give compound 45-d (2.0 g). LCMS (M+H)+: 474.
To a solution of compound 45-d (2.5 g, 5.28 mmol) in acetic acid (8 mL) was added H2O2 (11.27 g, 99.44 mmol) slowly, the mixture was then stirred at r.t. for 1 h. The reaction mixture was poured into saturated aqueous solution of Na2SO3 (200 mL) and extracted with ethyl acetate (3×250 mL). The combined organic layer was washed with brine (200 mL), dried over Na2SO4, filtered and concentrated. The residue was purified via column chromatography to afford compound 45-e (1.2 g). LCMS (M+H)+: 442.
To a solution of compound 45-e (100 mg, 0.23 mmol) in 1,4-dioxane (2 mL) was added 4-bromo-N-[(2,4-dimethoxyphenyl)methyl]-1-methyl-pyrazolo[4,3-c]pyridine-6-carboxamide (intermediate A2, 92 mg, 0.23 mmol), Pd(OAc)2 (15 mg, 0.07 mmol), CuI (86 mg, 0.45 mmol), PCy3HBF4 (33 mg, 0.09 mmol) and Cs2CO3 (73 mg, 0.23 mmol) under N2, the mixture was stirred at 140° C. for 12 hrs. The mixture was added in DCM/MeOH (30 mL/6 mL), then it was filtered. The filtrate was evaporated and purified via prep-TLC to obtain compound 45-f (20 mg). LCMS (M+H)+: 766.
A solution of compound 45-f (15.0 mg, 0.02 mmol) and MsOH (19 mg, 0.2 mmol) in HFIP (0.5 mL) was stirred at 30° C. for 3 hrs under N2 atmosphere. The reaction mixture was purified by reversed phase HPLC to obtain Example 45 (5.5 mg). LCMS (M+H)+: 406, 1H NMR (400 MHz, DMSO-d6) δ=15.37 (s, 1H), 8.93 (s, 1H), 8.89-8.84 (m, 1H), 8.47 (s, 1H), 8.02 (s, 1H), 7.88 (s, 1H), 4.57 (t, J=7.4 Hz, 2H), 4.23 (s, 3H), 2.63 (d, J=5.6 Hz, 2H), 2.14 (s, 3H), 1.73 (s, 3H).
The title compound was prepared according to the following scheme:
To a stirred mixture of 4-(benzyloxy)-3-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (31-e, 15.0 g, 47.1 mmol) in ACN (150 mL) was added NBS (16.8 g, 94.2 mmol) in portions. The resulting mixture was stirred for 2 h at 60° C. under nitrogen atmosphere. The mixture was allowed to cool down to 25° C. The resulting mixture was concentrated and the residue was purified via silica gel column chromatography to give compound 46-a (11.0 g). LCMS (M+H)+: 397.
To a stirred mixture of compound 46-a (11.0 g, 28 mmol) in THF (30 mL) was added tetrabutylammonium fluoride (150 mL, 150 mmol) (1 M in THF) dropwise. The resulting mixture was stirred for 15 h at 80° C. under nitrogen atmosphere. The mixture was allowed to cool down to 25° C. The resulting mixture was concentrated and purified via silica gel column chromatography to give compound 46-b (4.20 g). LCMS (M+H)+: 267.
To a solution of compound 46-b (500 mg, 1.87 mmol) in Toluene (5 mL) was added CMBP (1353 mg, 5.62 mmol) and 4-pyridineethanol (691 mg, 5.62 mmol). The mixture was stirred at 80° C. for 12 hrs. The mixture was concentrated and purified via prep-NPLC (Welch Ultimate XB—SiOH 250*70*10 um, Hexane-EtOH (0.1% FA, 10%˜50%) to give compound 46-c (250 mg) and compound 46-d (300 mg).
Compound 46-c, LCMS (M+H)+: 372, 1H NMR (400 MHz, CHLOROFORM-d) 6=8.61-8.46 (m, 2H), 7.43-7.33 (m, 5H), 7.09-7.00 (m, 2H), 4.91 (s, 2H), 4.28 (t, J=7.4 Hz, 2H), 3.12 (t, J=7.4 Hz, 2H), 2.10 (s, 3H).
Compound 46-d, LCMS (M+H)+: 372, 1H NMR (400 MHz, CHLOROFORM-d) 6=8.53 (br d, J=5.3 Hz, 2H), 7.37 (s, 5H), 7.08 (d, J=5.9 Hz, 2H), 4.93 (s, 2H), 4.15 (t, J=7.0 Hz, 2H), 3.17 (t, J=6.8 Hz, 2H), 1.68 (s, 3H).
A mixture of compound 46-c (119 mg, 0.32 mmol), N-[(2,4-dimethoxyphenyl)methyl]-1-[1-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (22-c, 150 mg, 0.29 mmol), Pd(OAc)2 (19 mg, 0.09 mmol), CuI (33 mg, 0.18 mmol), PPh3 (38 mg, 0.15 mmol), K2CO3 (121 mg, 0.88 mmol) in toluene (3 mL) was stirred at 120° C. for 12 hrs under N2 atmosphere. The reaction mixture was diluted with DCM (10 mL) and filtered. The filtrate was concentrated and purified via prep-TLC to afford compound 46-e (35 mg). LCMS (M+H)+: 805.
A solution of 1-[5-[4-benzyloxy-5-methyl-2-[2-(4-pyridyl)ethyl]pyrazol-3-yl]-1-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (46-e, 30 mg, 0.04 mmol) in HFIP (1.0 mL) was added MsOH (120 mg, 0.88 mmol). The mixture was stirred at 60° C. for 2 hrs and purified via reversed-phase HPLC to give Example 46 (11.2 mg). LCMS (M+H)+: 445, 1H NMR (400 MHz, DMSO-d6) δ=14.30-14.11 (m, 1H), 9.57 (d, J=1.0 Hz, 1H), 9.45-9.23 (m, 1H), 8.65 (d, J=6.4 Hz, 2H), 8.07 (s, 2H), 7.85 (br s, 1H), 7.79 (br d, J=5.8 Hz, 2H), 4.84 (br t, J=7.2 Hz, 2H), 3.38 (br t, J=7.0 Hz, 2H), 2.68 (s, 3H), 2.16 (s, 3H).
Example 47 was prepared in analogy to the preparation of Example 46 by using 3-(2-hydroxyethyl)pyridine instead of 4-pyridineethanol in step 3.
Example 47 (6.9 mg), LCMS (M+H)+: 445, 1H NMR (400 MHz, DMSO-d6) δ=9.65 (s, 1H), 8.87 (br s, 1H), 8.46 (br s, 1H), 8.37 (br d, J=4.4 Hz, 1H), 8.06 (s, 1H), 7.81 (br s, 1H), 7.69 (br d, J=7.6 Hz, 1H), 7.25 (dd, J=4.6, 7.8 Hz, 1H), 4.70 (br t, J=7.4 Hz, 2H), 3.10 (br t, J=7.4 Hz, 2H), 2.67 (s, 3H), 2.16 (s, 3H).
The title compound was prepared according to the following scheme:
To a solution of 4-benzyloxy-5-bromo-3-methyl-1-(2-phenylethyl)pyrazole (40-b, 90 mg, 0.24 mmol) in THF (2 mL) was added n-BuLi (2.5M, 0.15 mL, 0.36 mmol) dropwise at −78° C. under N2 atmosphere. The mixture was stirred at −78° C. for 1 h, and then triisopropyl borate (273 mg, 0.34 mL, 1.45 mmol) was added. The mixture was slowly warmed to r.t. and stirred for 2 hrs. The reaction mixture was poured into aq. NH4Cl (50 ml) and extracted with EA (3×60 mL). The combined organic layer was washed with brine, dried over Na2SO4 and concentrated to afford compound 48-a (80.0 mg). LCMS (M+H)+: 337.
To a stirred mixture of N-(2,4-dimethoxybenzyl)-5-methyl-1H-pyrazolo[3,4-c]pyridine-3-carboxamide (21-i, 800 mg, 2.45 mmol), 3,5-dibromo-1-methyl-1,2,4-triazole (1.76 g, 7.35 mmol) in DMF (15.0 mL) were added copper (I) iodide (233 mg, 1.23 mmol), 1,10-phenanthroline (441 mg, 2.45 mmol) and potassium carbonate (846 mg, 6.13 mmol) in portions. The resulting mixture was stirred for 16 hrs at 100° C. The mixture was cooled and diluted with ice water (60 mL), and extracted with EtOAc (3×80 mL). The combined organic layer was washed with brine (150 mL), dried over sodium sulfate and concentrated. The residue was purified via silica gel column chromatography to give compound 48-b (415 mg). LCMS (M+H)+: 485.
To a solution of 1-(5-bromo-2-methyl-1,2,4-triazol-3-yl)-N-[(2,4-dimethoxyphenyl) methyl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (48-b, 55 mg, 0.11 mmol) and compound 48-a (76 mg, 0.23 mmol) in 1,4-dioxane (3 mL) and water (0.3 mL) were added K2CO3 (280 mg, 2.04 mmol) and Pd(dppf)Cl2 (56 mg, 0.07 mmol). The mixture was stirred at 100° C. for 12 hrs under N2 atmosphere. The mixture was concentrated and purified via prep-TLC to obtain compound 48-c (30 mg). LCMS (M+H)+: 698.
A solution of compound 48-c (20 mg, 0.03 mmol) and MsOH (1.0 mL, 0.29 mmol) in HFIP (2.0 mL) was stirred at 30° C. for 3 hrs. The reaction mixture was purified via reversed phase HPLC to obtain Example 48 (10 mg). LCMS (M+H)+: 458, 1H NMR (400 MHz, DMSO-d6) δ=9.58 (s, 1H), 8.29 (br s, 1H), 8.07 (s, 1H), 8.05 (s, 1H), 7.91 (s, 1H), 7.25-7.11 (m, 5H), 4.58 (t, J=7.6 Hz, 2H), 4.29 (s, 3H), 3.04 (t, J=7.8 Hz, 2H), 2.68 (s, 3H), 2.14 (s, 3H).
Example 49 was prepared in analogy to the preparation of Example 48 by using compound 49-c instead of compound 40-b in step 1.
Compound 49-c prepared according to the following scheme:
To a solution of 4-benzyloxy-3-methyl-1H-pyrazole (31-f, 0.5 g, 2.66 mmol) in DMF (5 mL) was added Cs2CO3 (2.6 g, 7.97 mmol) and 1-bromobutane (1.09 g, 7.97 mmol). The mixture was stirred at 100° C. for 12 hrs. The reaction mixture was cooled and diluted with EtOAc (20 mL) and water (20 mL). The two-phase layer was separated, the aqueous phase was extracted with EtOAc (300 mL×2). All organic layers were combined, dried over Na2SO4 and concentrated to give compound 49-a (400 mg). LCMS (M+H)+: 245.
To a solution of compound 49-a (900 mg, 3.68 mmol) in THF (10 mL) was added NBS (786 mg, 4.42 mmol). The mixture was stirred at 20° C. for 1 h. The reaction mixture was concentrated and purified via prep-TLC to afford compound 49-b (200 mg). LCMS (M+H)+: 323, 1H NMR (400 MHz, CHLOROFORM-d) 6=7.46-7.30 (m, 5H), 4.93 (s, 2H), 4.03 (t, J=7.2 Hz, 2H), 2.10 (s, 3H), 1.83-1.72 (m, 2H), 1.37-1.26 (m, 2H), 0.94 (t, J=7.4 Hz, 3H).
To a solution of compound 49-b (46 mg, 0.14 mmol) in THF (2 mL) was added n-BuLi (2.5M, 0.09 mL, 0.21 mmol) dropwise at −78° C. under N2 atmosphere. The mixture was stirred at −78° C. for 0.5 h. Then triisopropyl borate (160 mg, 0.2 mL, 0.85 mmol) was added. The mixture was warmed to 25° C. and stirred for 0.5 h, then it was poured into NH4Cl aq. (50 ml) and extracted with EA (3×50 mL). The combined organic layer was dried over Na2SO4 and concentrated to give compound 49-c (50.0 mg). LCMS (M+H)+: 289.
Example 49, LCMS (M+H)+: 410, 1H NMR (400 MHz, DMSO-d6) δ=9.59 (d, J=0.8 Hz, 1H), 8.31 (s, 1H), 8.10 (s, 2H), 7.92 (s, 1H), 4.37 (t, J=7.2 Hz, 2H), 4.28 (s, 3H), 2.68 (s, 3H), 2.12 (s, 3H), 1.80-1.67 (m, 2H), 1.32-1.20 (m, 2H), 0.86 (t, J=7.2 Hz, 3H).
Example 50 was prepared in analogy to the preparation of Example 40 by using 4-methoxyphenethyl bromide instead of 1-bromo-2-phenylethane in step 1.
Example 50 (3.2 mg), LCMS (M+H)+: 474, 1H NMR (400 MHz, DMSO-d6) δ=14.16 (dd, J=2.6, 4.0 Hz, 1H), 9.62 (s, 1H), 9.32-9.15 (m, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.88 (br s, 1H), 7.15 (d, J=8.6 Hz, 2H), 6.73 (d, J=8.6 Hz, 2H), 4.66-4.58 (m, 2H), 3.67 (s, 3H), 3.00 (d, J=6.8 Hz, 2H), 2.68 (s, 3H), 2.18 (s, 3H).
The title compound was prepared according to the following scheme:
To a solution of 4-benzyloxy-5-bromo-3-methyl-1H-pyrazole (46-b, 300 mg, 1.12 mmol), 3-pentyn-1-ol (113 mg, 0.12 mL, 1.35 mmol) in toluene (5 mL) was added CMBP (541 mg, 2.25 mmol). The mixture was stirred at 80° C. for 16 hrs under N2 balloon. The mixture was concentrated and purified via prep-TLC to give compound 51-a (140 mg). LCMS (M+H)+: 332.
To a solution of 4-benzyloxy-5-bromo-3-methyl-1-pent-3-ynyl-pyrazole (51-a, 80 mg, 0.24 mmol) in THF (2 mL) was dropwise added n-BuLi (2.5 M, 0.14 mL, 0.36 mmol) at −70° C. under N2 atmosphere. The mixture was stirred at −70° C. for 1 h, and then triisopropyl borate (270 mg, 0.33 mL, 1.44 mmol) was added. The mixture was warmed to 20° C. slowly and stirred for 2 hrs. The reaction mixture was poured into saturated aqueous of NH4Cl (10 ml) at 0° C. and extracted with EtOAc (3×10 mL). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated to afford crude compound 51-b (60.0 mg). LCMS (M+H)+: 299.
A mixture of 3,5-dibromo-1H-1,2,4-triazole (30 g, 132 mmol), 4-methoxybenzylchloride (21.5 mL, 158.69 mmol), KI (10.98 g, 66 mmol), DIEA (43.7 mL, 264 mmol) in MeCN (200 mL) was stirred at 80° C. for 16 hrs under N2 balloon. The mixture was concentrated and purified via column chromatography to give compound 51-c (30 g). LCMS (M+H)+: 346, 1H NMR (400 MHz, DMSO-d6) δ=7.24 (d, J=8.6 Hz, 2H), 6.94 (d, J=8.6 Hz, 2H), 5.32 (s, 2H), 3.74 (s, 3H).
To a solution of N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-1H-pyrazolo[3,4-c]pyridine-3-carboxamide (21-i, 5.0 g, 15.32 mmol) and compound 51-c (7.97 g, 22.98 mmol) in DMF (30 mL) was added CuI (0.59 g, 3.06 mmol), 1-N,2-N-dimethylcyclohexane-1,2-diamine (435 mg, 3.06 mmol) and K3PO4 (6.5 g, 30.64 mmol). The mixture was stirred at 120° C. for 12 hrs under N2. The reaction mixture was poured into ice water (1000 mL) and extracted with ethyl acetate (3×500 mL). The organic layer was washed with water (2×500 mL) and brine (2×500 mL). The combined organic layer was dried over Na2SO4, filtered and concentrated. The residue was triturated with ethyl acetate (30 mL) and filtered to give compound 51-d (4.8 g). LCMS (M+H)+: 592, 1H NMR (400 MHz, DMSO-d6) δ=9.45 (s, 1H), 9.06 (t, J=5.6 Hz, 1H), 8.01 (s, 1H), 7.28 (d, J=8.4 Hz, 2H), 7.16 (d, J=8.4 Hz, 1H), 6.79 (d, J=8.4 Hz, 2H), 6.58 (s, 1H), 6.47 (d, J=8.2 Hz, 1H), 5.89 (s, 2H), 4.49 (d, J=5.6 Hz, 2H), 3.80 (s, 3H), 3.74 (s, 3H), 3.68 (s, 3H), 2.64 (s, 3H).
To a solution of compound 51-d (80 mg, 0.14 mmol) in 1,4-dioxane (5 mL) and water (0.500 mL) were added compound 51-b (60 mg, 0.2 mmol), K2CO3 (93 mg, 0.68 mmol) and Pd(dppf)Cl2·DCM (33 mg, 0.04 mmol). The mixture was stirred at 100° C. for 12 hrs under N2 atmosphere. The reaction mixture was concentrated, the residue was purified by flash silica gel chromatography to afford compound 51-e (50 mg). LCMS (M+H)+: 766.
To a solution of compound 51-e (90 mg, 0.12 mmol) in HFIP (2 mL) was added MsOH (112 mg, 1.18 mmol) at 25° C. The mixture was stirred at 25° C. for 1 h to give a red solution. The mixture was purified via reversed-phase column to give Example 51 (8 mg). LCMS (M+H)+: 406.1, 1H NMR (400 MHz, DMSO-d6) δ=14.25 (s, 1H), 9.71 (s, 1H), 9.35-9.09 (m, 1H), 8.06 (s, 2H), 7.78 (s, 1H), 4.59 (br t, J=6.8 Hz, 2H), 2.67 (s, 3H), 2.64-2.58 (m, 2H), 2.18 (s, 3H), 1.61 (s, 3H).
Example 52 was prepared in analogy to the preparation of Example 51 by using 1-(2-bromoethyl)-4-fluorobenzene instead of 3-pentyn-1-ol in step 1.
Example 52 (18.12 mg), LCMS (M+H)+: 462, 1H NMR (400 MHz, DMSO-d6) δ=9.60 (s, 1H), 8.03 (s, 1H), 7.99 (br s, 1H), 7.82 (br s, 1H), 7.27 (dd, J=5.6, 8.4 Hz, 2H), 6.98 (t, J=8.8 Hz, 2H), 4.73-4.59 (m, 2H), 3.06 (br t, J=7.6 Hz, 2H), 2.67 (s, 3H), 2.17 (s, 3H).
The title compound was prepared according to the following scheme:
To a solution of methyl 4-benzyloxy-3-methyl-1H-pyrazole-5-carboxylate (23-d, 2.0 g, 8.12 mmol) in DMF (20 mL) was added 1-bromo-2-phenylethane (2.25 g, 1.66 mL, 12.18 mmol) and K2CO3 (3.37 g, 24.36 mmol). The mixture was stirred at 80° C. for 12 hrs, and then it was concentrated. The residue was purified via column chromatography to give compound 53-a (1.7 g). LCMS (M+H)+: 351, 1H NMR (400 MHz, CHLOROFORM-d) δ=7.49-7.41 (m, 5H), 7.35 (d, J=7.4 Hz, 2H), 7.32-7.24 (m, 3H), 4.99 (s, 2H), 4.76-4.64 (m, 2H), 3.91 (s, 3H), 3.20-3.08 (m, 2H), 2.17 (s, 3H).
To a solution of compound 53-a (1.5 g, 4.28 mmol) in methanol (15 mL) was added hydrazine hydrate (2.19 g, 42.81 mmol) at 25° C. Then it was stirred at 80° C. for 4 hrs. The mixture was concentrated to give compound 53-b (1.4 g). LCMS (M+H)+: 351.
To a yellow solution of compound 53-b (1.5 g, 4.28 mmol) and DIPEA (2.12 mL, 12.84 mmol) in THF (20 mL) was added 4-methoxybenzyl isothiocyanate (1.53 g, 8.56 mmol) at 25° C. Then it was stirred at 30° C. for 3 hrs. The mixture was concentrated to obtained compound 53-c (2.2 g). LCMS (M+H)+: 530.
A yellow solution of compound 53-c (2.0 g, 3.78 mmol) in 3N NaOH (20 mL) was stirred at 100° C. for 12 hrs. The mixture was extracted with EA (3×100 mL). The organic layer was washed with water (3×200 mL) and brine (450 mL), dried over sodium sulfate and concentrated. The residue was purified via column chromatography to give compound 53-d (1.3 g) as a yellow oil. LCMS (M+H)+: 512.
To a solution of compound 53-d (1.3 g, 2.54 mmol) in acetic acid (4 mL) was added H2O2 (2.91 g, 25.68 mmol) slowly, then it was stirred at 25° C. for 1 h. The reaction mixture was poured into saturated solution of Na2SO3 (300 mL) and extracted with ethyl acetate (3×150 mL). The combined organic layer was washed with brine (300 mL), dried over Na2SO4 and concentrated. The residue was purified via column chromatography to afford compound 53-e (1.0 g). LCMS (M+H)+: 480.
To a solution of 1-bromo-N-[(2,4-dimethoxyphenyl)methyl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (Intermediate A1, 84 mg, 0.21 mmol) in 1,4-Dioxane (2 mL) was added Pd(OAc)2 (14 mg, 0.06 mmol), Cs2CO3 (67 mg, 0.21 mmol), PCy3HBF4 (30 mg, 0.08 mmol), CuI (79 mg, 0.42 mmol) and compound 53-e (0.1 g, 0.21 mmol) in one portion. The mixture was stirred at 140° C. for 12 hr in a sealed tube. (The reaction was set in glove box). The mixture was concentrated and purified via Prep-TLC to give compound 53-f (60 mg). LCMS (M+H)+: 804.
A solution of compound 53-f (40 mg, 0.05 mmol) and MsOH (49 mg, 0.5 mmol) in HFIP (2 mL) was stirred at 30° C. for 3 hrs. The reaction mixture was purified by reversed-phase HPLC to obtain Example 53 (18.0 mg). LCMS (M+H)+: 444, 1H NMR (400 MHz, DMSO-d6) δ=14.17-13.83 (m, 1H), 9.68 (d, J=1.4 Hz, 1H), 9.32-8.79 (m, 2H), 8.21-7.80 (m, 2H), 7.29-7.11 (m, 5H), 4.69 (d, J=1.8 Hz, 2H), 3.13-2.99 (m, 2H), 2.50 (s, 3H), 2.16 (s, 3H).
Example 54 was prepared in analogy to the preparation of Example 51 by using 1-(2-bromo-ethyl)-3-methoxy-benzene instead of 3-pentyn-1-ol in step 1.
Example 54 (6.9 mg), LCMS (M+H)+: 474, 1H NMR (400 MHz, DMSO-d6) δ=14.17 (br s, 1H), 9.63 (d, J=1.2 Hz, 1H), 9.34-9.15 (m, 1H), 8.08 (s, 1H), 7.98 (br d, J=0.9 Hz, 1H), 7.85 (br s, 1H), 7.09 (t, J=7.8 Hz, 1H), 6.79 (d, J=7.6 Hz, 1H), 6.75-6.67 (m, 2H), 4.69 (br t, J=7.6 Hz, 2H), 3.56 (s, 3H), 3.10-3.00 (m, 2H), 2.68 (s, 3H), 2.19 (s, 3H).
Example 55 was prepared in analogy to the preparation of Example 51 by using compound 55-b instead of compound 51-b in step 5.
The compound 55-b was prepared according to the following scheme:
To a solution of 4-benzyloxy-5-bromo-3-methyl-1H-pyrazole (46-b, 400.0 mg, 1.5 mmol) in DMF (1 mL) was added Cs2CO3 (1463.7 mg, 4.49 mmol) and 1-(2-bromoethyl)-3-fluoro-benzene (912 mg, 4.49 mmol). The mixture was stirred at 100° C. for 12 hrs. The reaction mixture was diluted with H2O (50 mL) and extracted with EtOAc (30 mL×3). The organic layer was washed with brine, dried with Na2SO4 and concentrated. The residue was purified via column chromatography and prep-HPLC to afford compound 55-a (130 mg). LCMS (M+H)+: 389, 1H NMR (400 MHz, CHLOROFORM-d) δ=7.43-7.33 (m, 5H), 7.26-7.21 (m, 1H), 6.98-6.87 (m, 2H), 6.82 (br d, J=10.0 Hz, 1H), 4.90 (s, 2H), 4.33-4.16 (m, 2H), 3.08 (t, J=7.6 Hz, 2H), 2.09 (s, 3H).
To a solution of 4-(benzyloxy)-5-bromo-1-(3-fluorophenethyl)-3-methyl-1H-pyrazole (55-a, 100 mg, 0.26 mmol) and 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (72 mg, 0.39 mmol) in THF (0.5 mL) and toluene (2.5 mL) was dropwise added n-BuLi (2.5 M, 0.12 mL, 0.31 mmol) at −78° C. under N2 atmosphere. The mixture was stirred at −78° C. for 1 h, then warmed to 20° C. slowly and stirred for 1 h. The mixture was poured into saturated aqueous solution of NH4Cl (10 ml) at 0° C. and extracted with EtOAc (3×10 mL). The organic layer was washed with brine and dried over anhydrous Na2SO4 and concentrated to afford crude compound 55-b (100 mg). The crude product was used to next step without purification. LCMS (M+H)+: 437.
Example 55 (18 mg), LCMS (M+H)+: 462, 1H NMR (400 MHz, DMSO-d6) δ=14.16 (br s, 1H), 9.61 (s, 1H), 9.24 (br s, 1H), 8.08 (s, 1H), 7.99 (br s, 1H), 7.83 (br s, 1H), 7.25-7.14 (m, 1H), 7.07-6.92 (m, 3H), 4.70 (br t, J=7.6 Hz, 2H), 3.10 (br t, J=7.6 Hz, 2H), 2.68 (s, 3H), 2.17 (s, 3H).
Example 56 was prepared in analogy to the preparation of Example 51 by using 2-fluorophenethyl alcohol instead of 3-pentyn-1-ol in step 1 and using 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane instead of triisopropyl borate in step 2.
Example 56 (35.3 mg), LCMS (M+H+): 462, 1H NMR (400 MHz, DMSO-d6) δ=14.35-13.88 (m, 1H), 9.63 (s, 1H), 9.03 (s, 1H), 8.11 (s, 1H), 8.02 (s, 1H), 7.81 (s, 1H), 7.29-7.15 (m, 2H), 7.04-6.92 (m, 2H), 4.72 (t, J=6.8 Hz, 2H), 3.11 (t, J=6.8 Hz, 2H), 2.69 (s, 3H), 2.17 (s, 3H).
Example 57 was prepared in analogy to the preparation of Example 53 by using compound 4-g instead of Intermediate A1 in step 6.
Example 57 (7.9 mg), LCMS (M+H)+: 461, 1H NMR (400 MHz, DMSO-d6) δ=14.37-13.69 (m, 1H), 9.63 (s, 1H), 9.22 (s, 1H), 9.21-8.78 (m, 1H), 7.92-7.74 (m, 1H), 7.56-7.37 (m, 1H), 7.23-7.16 (m, 5H), 4.69 (t, J=7.6 Hz, 2H), 3.08-3.02 (m, 2H), 2.49 (s, 3H), 2.16 (s, 3H).
The title compound was prepared according to the following scheme:
To a stirred mixture of ethyl 4-(3-(2,4-dimethoxybenzylcarbamoyl)-5-methyl-1H-pyrazolo[3,4-c]pyridin-1-yl)oxazole-5-carboxylate (21-j, 80 mg, 0.17 mmol) and 4-(benzyloxy)-5-bromo-3-methyl-1-propyl-1H-pyrazole (31-h, 106 mg, 0.34 mmol) in 1,4-dioxane (8 mL) was added copper(I) iodide (98 mg, 0.52 mmol), palladium (II) acetate (38 mg, 0.17 mmol), cesium carbonate (111 mg, 0.34 mmol) and tricyclohexylphosphonium tetrafluoroborate (44 mg, 0.12 mmol). The resulting mixture was stirred at 140° C. for 16 hrs under nitrogen atmosphere. The mixture was cooled down and concentrated. The residue was purified via silica gel column chromatography to give compound 58-a (40 mg). LCMS (M+H)+: 694.
To a stirred mixture of ethyl compound 58-a (35 mg, 0.05 mmol) in THF (1 mL) and methanol (1 mL) was added a solution of sodium hydroxide (22 mg, 0.55 mmol) in water (1 mL) dropwise at 0° C. The resulting mixture was stirred at 25° C. for 16 hrs and then acidified to pH=5 with HCl (1.0 M aq.) at 0° C. The suspension was filtered, the wet-cake was dried to give compound 58-b (25 mg). LCMS (M+H)+: 666.
To a stirred mixture of compound 58-b (20 mg, 0.03 mmol) in DMSO (4 mL) was added acetic acid (0.5 mg, 0.01 mmol) and silver carbonate (2.1 mg, 0.01 mmol). The resulting mixture was stirred at 85° C. for 3 hrs under nitrogen atmosphere. The mixture was poured into ice/water (10 mL) and extracted with EtOAc (3×10 mL). The combined organic layer was dried over anhydrous sodium sulfate and concentrated to give compound 58-c (18 mg). LCMS (M+H)+: 622.
To a stirred mixture of compound 58-c (18 mg, 0.03 mmol,) in 1,1,1,3,3,3-Hexafluoro-2-propanol (0.5 mL) was added methanesulfonic acid (31 mg, 0.32 mmol). The resulting mixture was stirred at 50° C. for 5 hrs under nitrogen atmosphere. The mixture was cooled down to room temperature. The residue was purified via reversed flash chromatography to give Example 58 (5.7 mg) as a white solid. LCMS (M+H)+: 382. 1H NMR (400 MHz, DMSO-d6) δ=9.57 (s, 1H), 8.99 (s, 1H), 8.66 (s, 1H), 8.00 (s, 2H), 7.78 (s, 1H), 4.46-4.42 (m, 2H), 2.65 (s, 3H), 2.16 (s, 3H), 1.84-1.82 (m, 2H), 0.92-0.89 (m, 3H).
Example 59 was prepared in analogy to the preparation of Example 58 by using compound 49-b instead of compound 31-h.
Example 59 (3.0 mg), LCMS (M+H)+: 396, 1H NMR (400 MHz, DMSO-d6) δ=9.55 (s, 1H), 9.01 (br s, 1H), 8.66 (s, 1H), 8.00 (s, 2H), 7.78 (s, 1H), 4.49-4.46 (m, 2H), 2.65 (s, 3H), 2.16 (s, 3H), 1.81-1.77 (m, 2H), 1.33-1.31 (m, 2H), 0.91-0.87 (m, 3H).
Example 60 was prepared in analogy to the preparation of Example 51 by using 3-(2-hydroxyethyl)benzonitrile instead of 3-pentyn-1-ol in step 1 and using 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane instead of triisopropyl borate in step 2.
Example 60 (30 mg), LCMS (M+H)+: 469, 1H NMR (400 MHz, DMSO-d6) δ=14.12 (br s, 1H), 9.60 (s, 1H), 9.25 (br s, 1H), 8.06 (s, 1H), 8.03 (br s, 1H), 7.80 (br s, 1H), 7.61-7.55 (m, 2H), 7.53 (br d, J=7.8 Hz, 1H), 7.36 (t, J=7.8 Hz, 1H), 4.75 (br t, J=7.2 Hz, 2H), 3.16 (br t, J=7.2 Hz, 2H), 2.68 (s, 3H), 2.18 (s, 3H).
Example 61 was prepared in analogy to the preparation of Example 51 by using 4-(2-hydroxyethyl)benzonitrile instead of 3-pentyn-1-ol in step 1 and using 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane instead of triisopropyl borate in step 2.
Example 61 (17.6 mg), LCMS (M+H)+: 469, 1H NMR (400 MHz, DMSO-d6) δ=14.14 (s, 1H), 9.57 (d, J=1.0 Hz, 1H), 9.42-9.15 (m, 1H), 8.06 (s, 2H), 7.86 (s, 1H), 7.61 (d, J=8.2 Hz, 2H), 7.42 (d, J=8.2 Hz, 2H), 4.72 (t, J=7.2 Hz, 2H), 3.17 (t, J=7.2 Hz, 2H), 2.68 (s, 3H), 2.16 (s, 3H).
Example 62 was prepared in analogy to the preparation of Example 51 by using methyl 4-(2-hydroxyethyl)benzoate instead of 3-pentyn-1-ol in step 1 and using 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane instead of triisopropyl borate in step 2.
Example 62 (14 mg), LCMS (M+H)+: 502, 1H NMR (400 MHz, DMSO-d6) δ=14.13 (s, 1H), 9.57 (d, J=1.0 Hz, 1H), 9.23 (s, 1H), 8.09-7.99 (m, 2H), 7.83 (br s, 1H), 7.72 (d, J=8.4 Hz, 2H), 7.32 (d, J=8.0 Hz, 2H), 4.73 (br t, J=7.2 Hz, 2H), 3.79 (s, 3H), 3.17-3.12 (m, 2H), 2.67 (s, 3H), 2.17 (s, 3H).
The title compound was prepared according to the following scheme:
2-[4-benzyloxy-3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl]ethoxy-tert-butyl-dimethyl-silane (compound 63-a) was prepared in analogy to the preparation of compound 49-c by using (2-bromoethoxy)-tert-butyldimethylsilane instead of 1-bromobutane in step 1 and using reagent 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane instead of triisopropyl borate in step 3.
To a solution of compound 63-a (1.3 g, 2.75 mmol) in 1,4-dioxane (10 mL) and water (1 mL) was added 1-[5-bromo-1-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (51-d, 1.0 g, 1.69 mmol), K2CO3 (1.2 g, 8.44 mmol) and Pd(dppf)Cl2·DCM (413 mg, 0.51 mmol). The mixture was stirred at 100° C. for 12 hrs under N2 atmosphere. It was then cooled and poured into water (100 mL). The suspension was extracted with EtOAc (100 mL×2), the organic phase was washed with brine (20 mL×2), dried over Na2SO4 and concentrated. The residue was purified via column chromatography to give compound 63-b (1.1 g). LCMS (M+H)+: 858.
To a solution of 1-[5-[4-benzyloxy-2-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-5-methyl-pyrazol-3-yl]-2-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (63-b, 1.1 g, 1.28 mmol) in methanol (10 mL) was added NH4F (0.95 g, 25.6 mmol). The mixture was stirred at 70° C. for 1 h. The mixture was cooled, filtered and washed with MeOH (40 mL×2). The filtrate was concentrated and purified with column chromatography to give compound 63-c (750 mg). LCMS (M+H)+: 744.
To a solution of compound 63-c (300 mg, 0.40 mmol) in DCM (10 mL) was added Dess-Martin periodinane (684 mg, 1.61 mmol). The mixture was stirred at 25° C. for 2 hrs. It was then poured into a mixture of saturated Na2SO3 solution (30 mL) and saturated NaHCO3 solution (30 mL). The mixture was stirred for 20 mins and extracted with DCM (30 mL). The organic layer was dried over Na2SO4, filtered and concentrated to give compound 63-d (300 mg). LCMS (M+H)+: 742.
To a solution of compound 63-d (300 mg, 0.40 mmol) in DMF (1 mL) was added 2-(triphenylphosponio)difluoroacetate (582 mg, 1.62 mmol). The mixture was stirred at 80° C. for 12 hrs. The mixture was poured into water (30 mL), and extracted with EtOAc (30 mL×2), the organic phase was washed with brine (10 mL×3), dried over Na2SO4, and concentrated. The residue was purified via prep-TLC to give compound 63-e (60 mg). LCMS (M+H)+: 776.
To a solution of compound 63-e (55 mg, 0.07 mmol) in HFIP (0.3 mL) was added MsOH (136 mg, 1.42 mmol). The mixture was stirred at 50° C. for 0.5 h. The mixture was then directly purified via prep-HPLC to give Example 63 (30 mg). LCMS (M+H)+: 416, 1H NMR (DMSO-d6, 400 MHz) δ=14.28 (br s, 1H), 9.62 (s, 1H), 9.34 (br s, 1H), 8.05 (s, 2H), 7.78 (br s, 1H), 5.13 (br d, J=7.6 Hz, 2H), 5.02-4.90 (m, 1H), 2.66 (s, 3H), 2.18 (s, 3H).
Example 64 was prepared in analogy to the preparation of Example 51 by using compound 64-a instead of 3-pentyn-1-ol in step 1 and using 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane instead of triisopropyl borate in step 2.
The compound 64-a was prepared according to the following scheme:
To a solution of 2-[4-(methoxymethyl)phenyl]acetic acid (800 mg, 4.44 mmol) in THF (20 mL) was added LiAlH4 (505 mg, 13.32 mmol) at 0° C., then the mixture was stirred at 60° C. for 0.5 h. The reaction mixture was added into 1M aqueous solution of HCl (50 mL), extracted with EtOAc (50 mL×3). The combined organic phase was dried over Na2SO4, filtered and concentrated to give compound 64-a (650 mg). LCMS (M+H)+: 415, 1H NMR (400 MHz, CHLOROFORM-d) δ=7.34-7.30 (m, 2H), 7.26-7.21 (m, 2H), 4.46 (s, 2H), 3.88 (t, J=6.8 Hz, 2H), 3.41 (s, 3H), 2.89 (t, J=6.4 Hz, 2H).
Example 64 (30 mg), LCMS (M+H)+: 488, 1H NMR (400 MHz, DMSO-d6) δ=14.16 (br s, 1H), 9.62 (d, J=1.0 Hz, 1H), 9.23 (br s, 1H), 8.08 (s, 1H), 8.00 (br s, 1H), 7.86 (br s, 1H), 7.23-7.18 (d, J=8.0 Hz, 2H), 7.11 (d, J=8.0 Hz, 2H), 4.68 (br t, J=7.6 Hz, 2H), 4.32 (s, 2H), 3.23 (s, 3H), 3.10-3.03 (br t, J=6.8 Hz, 2H), 2.69 (s, 3H), 2.19 (s, 3H).
Example 65 was prepared in analogy to the preparation of Example 51 by using compound 65-c instead of 3-pentyn-1-ol in step 1 and using 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane instead of triisopropyl borate in step 2.
The compound 65-c was prepared according to the following scheme:
To a solution of methyl 3-methylphenylacetate (9.0 g, 54.81 mmol) in carbon tetrachloride (1 mL) was added BPO (1.33 g, 5.48 mmol), and NBS (9.76 g, 54.81 mmol). The reaction mixture was stirred at 90° C. for 18 hrs under N2. The reaction mixture was poured into water (100 mL), extracted with DCM (200 mL×2). The combined organic layer was washed with brine (100 mL), dried over Na2SO4, and filtered. The filtrate was concentrated and purified via flash chromatography on silica gel to afford compound 65-a (5.0 g). LCMS (M+H)+: 243, 1H NMR (400 MHz, DMSO-d6) δ=7.43-7.29 (m, 3H), 7.27-7.18 (m, 1H), 4.70 (s, 2H), 3.70 (s, 2H), 3.62 (s, 3H).
To a solution of compound 65-a (5.0 g, 20.6 mmol) in methanol (50 mL) was added NaOMe (5.0 mL, 25.0 mmol) at 0° C. The reaction mixture was stirred at 25° C. for 18 hrs and poured into Sat. NH4Cl (50 mL), it was then extracted with EA (50 mL×2). The combined organic layer was washed with brine (20 mL), dried over Na2SO4 and concentrated. The residue was purified via reversed-phase column to give compound 65-b (1.5 g). LCMS (M-CH3O)+: 163.
To a solution of compound 65-b (1.5 g, 7.7 mmol) in THF (15 mL) was added LiAlH4 (2.5 M, 3.86 mL, 9.64 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 1 h and poured into 1 N aqueous solution of HCl (20 mL). It was then extracted with EA (30 mL×2). The combined organic layer was washed with brine (20 mL), dried over Na2SO4, and concentrated to give a residue. The residue was purified via column to give compound 65-c (600 mg). LCMS (M-CH3O)+: 135.
Example 65 (32.3 mg), LCMS (M+H)+: 488, 1H NMR (400 MHz, DMSO-d6) δ=14.16 (br d, J=2.6 Hz, 1H), 9.65 (d, J=1.0 Hz, 1H), 9.47-8.96 (m, 1H), 8.12 (s, 1H), 8.07 (br s, 1H), 7.85 (br s, 1H), 7.20-7.04 (m, 4H), 4.75-4.65 (m, 2H), 4.17 (s, 2H), 3.09 (s, 3H), 3.07 (br s, 2H), 2.70 (s, 3H), 2.19 (s, 3H).
The title compound was prepared according to the following scheme:
To a solution of 1-(3-(1-(3,3-difluoroallyl)-4-hydroxy-3-methyl-1H-pyrazol-5-yl)-1H-1,2,4-triazol-5-yl)-5-methyl-1H-pyrazolo[3,4-c]pyridine-3-carboxamide (Example 63, 20 mg, 0.05 mmol) in THF (2 mL) and EtOAc (2 mL) was added Pd/C (20 mg) at 20° C. under N2 atmosphere. The mixture was degassed and purged with H2 for three times, then it was stirred at 20° C. for 10 mins with H2 balloon. The mixture was purified via prep-HPLC to give Example 66 (5 mg) as a yellow solid. LCMS (M+H)+: 418, 1H NMR (DMSO-d6, 400 MHz) δ=14.34-14.17 (m, 1H), 9.62 (d, J=0.8 Hz, 1H), 9.43-9.20 (m, 1H), 8.15-7.95 (m, 2H), 7.79 (br s, 1H), 6.34-5.98 (m, 1H), 4.66 (t, J=7.2 Hz, 2H), 2.67 (s, 3H), 2.43-2.35 (m, 2H), 2.18 (s, 3H).
Example 67 was prepared in analogy to the preparation of Example 58 by using compound 67-a instead of compound 31-h.
The compound 67-a was prepared according to the following scheme:
To a stirred mixture of 4-(benzyloxy)-5-bromo-3-methyl-1H-pyrazole (46-b, 1.50 g, 5.62 mmol), 3,3-difluoropropan-1-ol (863 mg, 8.98 mmol) and triphenylphosphine (1.91 g, 7.30 mmol) in THF (25 mL) was added diisopropyl azodicarboxylate (1.70 g, 8.42 mmol) dropwise at 0° C. The resulting mixture was stirred at 40° C. for 4 hrs. It was then cooled and poured into ice/water (50 mL) slowly. The mixture was extracted with EtOAc (3×50 mL), the organic layer was dried and concentrated. The residue was purified via silica gel column chromatography to give compound 67-a (800 mg). LCMS (M+H)+: 345. The structure was confirmed with 1H NMR and Nuclear Overhauser Effect Spectroscopy (NOESY) (no correlation between Ha and Hb was observed).
Example 67 (8.8 mg), LCMS (M+H)+: 418. 1H NMR (400 MHz, DMSO-d6) δ=9.54 (s, 1H), 9.19 (br s, 1H), 8.67 (s, 1H), 7.99 (s, 2H), 7.77 (s, 1H), 6.31-6.01 (m, 1H), 4.69-4.66 (m, 2H), 2.64 (s, 3H), 2.46-2.29 (m, 2H), 2.16 (s, 3H).
The title compound was prepared according to the following scheme:
A solution of 1-[3-[4-hydroxy-2-[2-[4-(methoxymethyl)phenyl]ethyl]-5-methyl-pyrazol-3-yl]-1H-1,2,4-triazol-5-yl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (Example 64, 10 mg, 0.02 mmol) in trifluoroacetic acid (2.0 mL, 25.96 mmol) and water (0.2 mL) was stirred at 100° C. for 6 hrs. The mixture was filtered, the filtrate was purified via prep-HPLC to afford Example 68 (1.4 mg), LCMS (M+H)+: 474, 1H NMR (400 MHz, DMSO-d6) δ=14.16 (br s, 1H), 9.63 (s, 1H), 9.22 (s, 1H), 8.06 (s, 1H), 7.98 (br s, 1H), 7.84 (br s, 1H), 7.21-7.16 (m, 2H), 7.15-7.10 (m, 2H), 4.65 (br dd, J=7.6, 8.4 Hz, 2H), 4.41 (s, 2H), 3.08-3.02 (m, 2H), 2.68 (s, 3H), 2.18 (s, 3H).
Example 69 was prepared in analogy to the preparation of Example 53 by using 4-methoxyphenethyl bromide instead of 1-bromo-2-phenylethane in step 1 and using compound 4-g instead of Intermediate A1 in step 6.
Example 69 (6.2 mg), LCMS (M+H)+: 491, 1H NMR (400 MHz, DMSO-d6) δ=14.19-13.59 (m, 1H), 9.61 (s, 1H), 9.22 (s, 1H), 9.19-8.91 (m, 1H), 7.93-7.69 (m, 1H), 7.60-7.37 (m, 1H), 7.10 (d, J=8.6 Hz, 2H), 6.76 (d, J=8.6 Hz, 2H), 4.69-4.62 (m, 2H), 3.68 (s, 3H), 3.00-2.95 (m, 2H), 2.49-2.48 (m, 3H), 2.16 (s, 3H).
Example 70 was prepared in analogy to the preparation of Example 53 by using 4-methoxyphenethyl bromide instead of 1-bromo-2-phenylethane in step 1.
Example 70 (20 mg), LCMS (M+H)+: 474.3, 1H NMR (400 MHz, DMSO-d6+D2O) δ=9.57 (s, 1H), 9.02 (s, 1H), 7.00 (d, J=8.4 Hz, 2H), 6.66 (d, J=8.4 Hz, 2H), 4.60 (t, J=7.2 Hz, 2H), 3.59 (s, 3H), 2.91 (t, J=7.2 Hz, 2H), 2.45 (s, 3H), 2.12 (s, 3H).
The title compound was prepared according to the following scheme:
To a solution of 4-(2-bromoethyl)benzoic acid (20.0 g, 87.3 mmol) in THF (200 mL) was added BH3·THF (1.0 M, 174.6 mL, 174.6 mmol) at 0° C. The reaction mixture warmed to 25° C. and stirred for 2 hrs. It was then poured into 1 N HCl (100 mL) and extracted with EA (200 mL×2). The combined layer was washed with brine (20 mL), dried over Na2SO4 and concentrated to give compound 71-a (18.0 g) as light yellow solid. LCMS (M+H−18)+: 197.
To a solution of [4-(2-bromoethyl)phenyl]methanol 71-a (8.0 g, 37.2 mmol), Et3N (14.3 mL, 111.6 mmol) in DCM (100 mL) was added TBSCl (8.41 g, 55.8 mmol) at 0° C. The reaction mixture warmed to 25° C. and stirred at for 18 hrs. Then it was poured into water (100 mL) and extracted with DCM (100 mL×2). The combined layer was washed with brine (100 mL), dried over Na2SO4, and concentrated to give an oil. The residue was purified via column to give 71-b (8.0 g). LCMS (M+H—C6H15SiO)+: 197.
Compound 71-c was prepared in analogy to the preparation of Example 53 by using compound 71-b instead of 1-bromo-2-phenylethane in step 1.
To a yellow solution of compound 71-c (1.5 g, 2.9 mmol), imidazole (400 mg, 5.9 mmol) in DCM (20 mL) was added TBSCl (2.96 g, 5.9 mmol) at 25° C. The mixture was stirred for 2 hrs and concentrated. The residue was purified via column chromatography to give compound 71-d (1.2 g). LCMS (M+H)+: 624.
A suspension of compound 71-d (300 mg, 0.48 mmol), 1-bromo-N-[(2,4-dimethoxyphenyl)methyl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (Intermediate A1, 194 mg, 0.48 mmol), Pd(OAc)2 (32.4 mg, 0.14 mmol), PivOH (29 mg, 0.29 mmol), catacxiuma (52 mg, 0.14 mmol), K2CO3 (199 mg, 1.44 mmol) in toluene (2 mL) was stirred at 120° C. for 12 hrs under N2 atmosphere. The mixture was filtered and washed with DCM/MeOH(V/V=20/1, mL). The filtrate was concentrated and purified by prep-TLC to afford compound 71-e (55 mg). LCMS (M+H)+: 948.
A solution of compound 71-e (50.0 mg, 0.05 mmol) in TFA (3 mL) was stirred at 60° C. for 12 hrs under N2 atmosphere. The mixture was concentrated and purified via prep-HPLC to give Example 71 (6.2 mg). LCMS (M+H)+: 474, 1H NMR (400 MHz, DMSO-d6+D2O) δ=9.68 (s, 1H), 9.10 (s, 1H), 7.36-6.83 (m, 4H), 4.66 (br s, 2H), 4.41 (s, 2H), 3.04 (br t, J=7.8 Hz, 2H), 2.50-2.50 (m, 3H), 2.16 (s, 3H).
Example 72 was prepared in analogy to the preparation of Example 53 by using 2-(3-pyridyl)ethanol instead of 1-bromo-2-phenylethane in step 1 (reaction condition: CMBP, toluene 80° C. instead of K2CO3, DMF 80° C.).
Example 72 (18.7 mg), LCMS (M+H)+: 445, 1H NMR (400 MHz, DMSO+D2O) δ=9.65 (s, 1H), 9.12 (s, 1H), 9.05-8.81 (m, 1H), 8.35 (br d, J=3.8 Hz, 1H), 7.63 (br d, J=7.4 Hz, 1H), 7.24 (dd, J=4.8, 7.6 Hz, 1H), 4.69 (br t, J=7.2 Hz, 2H), 3.09 (br t, J=7.2 Hz, 2H), 2.49 (br s, 3H), 2.15 (s, 3H).
Example 73 was prepared in analogy to the preparation of Example 53 by using 2-(3-pyridyl)ethanol instead of 1-bromo-2-phenylethane in step 1 (reaction condition: CMBP, toluene 80° C. instead of K2CO3, DMF 80° C.) and using compound 4-g instead of Intermediate A1 in step 6.
Example 73 (8.91 mg), LCMS (M+H)+: 462, 1H NMR (400 MHz, DMSO-d6+D2O) δ=9.56 (s, 1H), 9.17 (s, 1H), 8.34-8.23 (m, 2H), 7.52 (br d, J=8.4 Hz, 1H), 7.18 (dd, J=4.8, 7.8 Hz, 1H), 4.73 (br t, J=7.2 Hz, 2H), 3.09 (br t, J=7.2 Hz, 2H), 2.46 (s, 3H), 2.13 (s, 3H).
Example 74 was prepared in analogy to the preparation of Example 58 by using compound 74-a instead of compound 31-h.
The compound 74-a was prepared according to the following scheme:
To a stirred mixture of 4-(benzyloxy)-5-bromo-3-methyl-1H-pyrazole (46-b, 500 mg, 1.90 mmol), 2-(pyridin-3-yl)ethanol (345 mg, 2.80 mmol) and triphenylphosphine (638 mg, 2.40 mmol) in THF (8 mL) was added diisopropyl azodicarboxylate (492 mg, 2.40 mmol) dropwise at 0° C. The resulting mixture was stirred at 40° C. for 4 hrs. The mixture was cooled and concentrated. The residue was purified via silica gel column to give compound 74-a (280 mg). LCMS (M+H)+: 372.
Example 74 (15 mg). LCMS (M+H)+: 445. 1H NMR (400 MHz, DMSO-d6) δ=9.50 (s, 1H), 9.07 (br s, 1H), 8.70 (s, 1H), 8.64 (s, 2H), 8.13 (s, 1H), 8.07 (s, 2H), 7.83 (s, 1H), 7.74 (s, 1H), 4.84-4.81 (m, 2H), 3.35-3.32 (m, 2H), 2.67 (s, 3H), 2.13 (s, 3H).
Example 75 was prepared in analogy to the preparation of Example 58 by using compound 75-a instead of compound 31-h.
Compound 75-a was prepared in analogy to the preparation of compound 74-a by using 2-(3-methoxyphenyl)ethanol instead of 2-(pyridin-3-yl)ethanol.
Example 75 (8.7 mg), LCMS (M+H)+: 474. 1H NMR (400 MHz, DMSO-d6) δ=9.52 (s, 1H), 8.96 (s, 1H), 8.65 (s, 1H), 8.01 (s, 1H), 7.98 (s, 1H), 7.79 (s, 1H), 7.07-7.03 (m, 1H), 6.71-6.65 (m, 3H), 4.72-4.68 (m, 2H), 3.53 (s, 3H), 3.06-3.02 (m, 2H), 2.65 (s, 3H), 2.17 (s, 3H).
Example 76 was prepared in analogy to the preparation of Example 53 by using 1-bromobutane instead of 1-bromo-2-phenylethane in step 1 and using compound 4-g instead of Intermediate A1 in step 6.
Example 76 (12.3 mg), LCMS (M+H)+: 413, 1H NMR (400 MHz, DMSO-d6) δ=14.24-13.68 (m, 1H), 9.63 (s, 1H), 9.27-9.18 (m, 1H), 9.16-8.82 (m, 1H), 7.93-7.70 (m, 1H), 7.59-7.35 (m, 1H), 4.47 (t, J=7.0 Hz, 2H), 2.48 (s, 3H), 2.14 (s, 3H), 1.75 (br t, J=7.4 Hz, 2H), 1.34-1.23 (m, 2H), 0.87 (t, J=7.4 Hz, 3H).
Example 77 was prepared in analogy to the preparation of Example 53 by using 1-bromobutane instead of 1-bromo-2-phenylethane in step 1.
Example 77 (12.1 mg), LCMS (M+H)+): 396, 1H NMR (400 MHz, DMSO-d6) δ=14.32-13.78 (m, 1H), 9.69 (d, J=1.4 Hz, 1H), 9.10 (s, 1H), 9.08-8.75 (m, 1H), 8.22-7.79 (m, 2H), 4.46 (t, J=6.6 Hz, 2H), 2.49-2.48 (m, 3H), 2.15 (s, 3H), 1.75 (br t, J=7.4 Hz, 2H), 1.32-1.23 (m, 2H), 0.86 (t, J=7.4 Hz, 3H).
Example 78 was prepared in analogy to the preparation of Example 58 by using compound 78-a instead of compound 31-h.
Compound 78-a was prepared in analogy to the preparation of compound 74-a by using 2-(4-methoxyphenyl)ethanol instead of 2-(pyridin-3-yl)ethanol.
Example 78 (2.3 mg), LCMS (M+H)+: 474. 1H NMR (400 MHz, DMSO-d6) δ=9.52 (s, 1H), 8.95 (s, 1H), 8.67 (s, 1H), 8.03 (d, J=5.6 Hz, 1H), 7.99 (s, 1H), 7.80 (s, 1H), 7.06 (d, J=8.4, 2H), 6.70 (d, J=8.4, 2H), 4.67-4.63 (m, 2H), 3.65 (s, 3H), 3.02-2.98 (m, 2H), 2.66 (s, 3H), 2.17 (s, 3H).
Example 79 was prepared in analogy to the preparation of Example 53 by using 1-propanol instead of 1-bromo-2-phenylethane in step 1 (reaction condition: CMBP, toluene 80° C. instead of K2CO3, DMF 80° C.) and using compound 4-g instead of Intermediate A1 in step 6.
Example 79 (18.4 mg), LCMS (M+H)+: 399, 1H NMR (400 MHz, DMSO-d6) δ=14.43-13.53 (m, 1H), 9.70-9.58 (m, 1H), 9.23 (s, 1H), 9.14-8.81 (m, 1H), 7.87-7.74 (m, 1H), 7.52-7.39 (m, 1H), 4.43 (br t, J=7.2 Hz, 2H), 2.48 (s, 3H), 2.16 (s, 3H), 1.85-1.74 (m, 2H), 0.88 (t, J=7.4 Hz, 3H).
Example 80 was prepared in analogy to the preparation of Example 53 by using 1-propanol instead of 1-bromo-2-phenylethane in step 1 (reaction condition: CMBP, toluene 80° C. instead of K2CO3, DMF 80° C.).
Example 80 (4.9 mg) as yellow solid. LCMS (M+H)+: 382, 1H NMR (400 MHz, DMSO-d6+D2O=9.67 (s, 1H), 9.08 (s, 1H), 4.45-4.32 (m, 2H), 2.47 (br s, 3H), 2.14 (br s, 3H), 1.81-1.72 (m, 2H), 0.83 (t, J=7.4 Hz, 3H).
Example 81 was prepared in analogy to the preparation of Example 51 by using compound 81-b instead of 3-pentyn-1-ol in step 1 and using 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane instead of triisopropyl borate in step 2.
The compound 81-b was prepared according to the following scheme:
To a solution of tert-butyl-but-3-ynoxy-dimethyl-silane (5.0 g, 27.1 mmol) in THF (25 mL) was added n-BuLi (13.0 mL, 32.6 mmol) drop-wise at −78° C. under N2 atmosphere. The mixture was stirred at −78° C. for 0.5 h, and then bromomethyl methyl ether (4.41 g, 35.3 mmol) in THF (5 mL) was added at −78° C. The mixture was stirred at −78° C. for another 0.5 h. The reaction mixture was poured into water (100 ml) and extracted with ethyl acetate (3×250 mL). The combined organic layer was washed with brine and dried over Na2SO4. It was then concentrated and purified by column chromatography to give compound 81-a (5.6 g). LCMS (M+H)+: 229, 1H NMR (400 MHz, CHLOROFORM-d) δ=4.08 (d, J=1.8 Hz, 2H), 3.74 (t, J=7.2 Hz, 2H), 3.38 (s, 3H), 2.46 (d, J=1.8 Hz, 2H), 0.91 (s, 9H), 0.08 (s, 6H).
To a solution of compound 81-a (5.0 g, 21.9 mmol) in methanol (30 mL) was added NH4F (24.33 g, 656.7 mmol). The mixture was stirred at 60° C. for 1 h. The mixture was filtered and washed with DCM (200 mL×2). The filtrate was concentrated and purified by column chromatography to give compound 81-b (2.4 g). LCMS (M+H)+: 115, 1H NMR (400 MHz, CHLOROFORM-d) δ=4.11 (d, J=1.8 Hz, 2H), 3.86-3.62 (m, 2H), 3.39 (d, J=1.4 Hz, 3H), 2.67-2.42 (m, 2H).
Example 81 (8.6 mg), LCMS (M+H)+: 436, 1H NMR (400 MHz, DMSO-d6) δ=14.21 (s, 1H), 9.68 (s, 1H), 9.34-9.18 (m, 1H), 8.06 (d, J=5.0 Hz, 2H), 7.78 (d, J=3.2 Hz, 1H), 4.65 (t, J=6.8 Hz, 2H), 3.96 (s, 2H), 3.10 (s, 3H), 2.76 (t, J=6.8 Hz, 2H), 2.66 (s, 3H), 2.18 (s, 3H).
Example 82 was prepared in analogy to the preparation of Example 53 by using Intermediate A2 instead of Intermediate A1 in step 6.
Example 82 (8.4 mg), LCMS (M+H)+: 444. 1H NMR (400 MHz, DMSO-d6) δ=15.33-15.16 (br s, 1H), 8.82 (br s, 1H), 8.79 (s, 2H), 8.46 (s, 1H), 7.88 (s, 1H), 7.26-7.15 (m, 5H), 4.70-4.66 (m, 2H), 4.22 (s, 3H), 3.09-3.05 (m, 2H), 2.16 (s, 3H).
The title compound was prepared according to the following scheme:
To a stirred mixture of ethyl 2-(4-benzyloxy-5-methyl-2-propyl-pyrazol-3-yl)-4-[3-[(2,4-dimethoxyphenyl)methylcarbamoyl]-5-methyl-pyrazolo[3,4-c]pyridin-1-yl]oxazole-5-carboxylate (compound 58-a, 200 mg, 0.29 mmol) in THF (15 mL) was added LiBH3NMe2 (0.49 mL, 0.49 mmol) (1 M in THF) dropwise at 0° C. The reaction was stirred at 25° C. for 1 h. The mixture was concentrated and purified via silica gel column chromatography to give compound 83-a (100 mg). LCMS (M+H)+: 652.
A mixture of 1-[2-(4-benzyloxy-5-methyl-2-propyl-pyrazol-3-yl)-5-(hydroxymethyl)oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-5-methyl-pyrazolo[3,4-c]pyridine-3-carboxamide (compound 83-a, 20 mg, 0.03 mmol) and MsOH (23 mg, 0.25 mmol) in HFIP (2 mL) was stirred for 3 hrs at 50° C. under nitrogen atmosphere. The mixture was cooled to room temperature and purified via reversed phase HPLC to give Example 84 (3.5 mg). LCMS (M+H)+: 412, 1H NMR (400 MHz, DMSO-d6) δ=9.52 (s, 1H), 8.90 (s, 1H), 8.12 (s, 1H), 8.03 (s, 1H), 7.78 (s, 1H), 5.51 (br s, 1H), 4.92 (s, 2H), 4.43-4.40 (m, 2H), 2.65 (s, 3H), 2.16 (s, 3H), 1.83-1.78 (m, 2H), 0.90-0.86 (m, 3H).
Example 84 was prepared in analogy to the preparation of Example 48 by using compound 31-h instead of compound 40-b and using 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane instead of triisopropyl borate in step 1.
Example 84 (58 mg), LCMS (M+H)+: 396, 1H NMR (400 MHz, DMSO-d6) δ=9.59 (d, J=1.0 Hz, 1H), 8.30 (s, 1H), 8.09 (s, 2H), 7.92 (s, 1H), 4.33 (t, J=7.2 Hz, 2H), 4.28 (s, 3H), 2.68 (s, 3H), 2.12 (s, 3H), 1.84-1.71 (m, 2H), 0.85 (t, J=7.4 Hz, 3H)
The title compound was prepared according to the following scheme:
To a solution of ethyl 4-bromooxazole-5-carboxylate (896 mg, 4.07 mmol), 1-bromo-N-[(3,4-dimethylphenyl)methyl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (Intermediate A1, 1500 mg, 3.7 mmol) in toluene (1 mL) and water (0.1 mL) was added K3PO4 (2357 mg, 11.1 mmol), B2Pin2 (1880 mg, 7.4 mmol), Pd(dppf)Cl2 (541 mg, 0.74 mmol) at 25° C. The mixture was degassed and purged with N2 three times. The mixture was heated to 80° C. and stirred for 18 hrs. The mixture was concentrated and purified via flash chromatography on silica gel to afford compound 85-a (500 mg). LCMS (M+H)+: 466.
A mixture of ethyl 4-[3-[(2,4-dimethoxyphenyl)methylcarbamoyl]-6-methyl-imidazo[1,5-a]pyrazin-1-yl]oxazole-5-carboxylate (85-a, 200 mg, 0.43 mmol), 4-benzyloxy-5-bromo-3-methyl-1-propyl-pyrazole (31-h, 132 mg, 0.43 mmol), Pd(OAc)2 (29 mg, 0.13 mmol), PivOH (26 mg, 0.26 mmol), catacxiuma (46 mg, 0.13 mmol), K2CO3 (178 mg, 1.29 mmol) in toluene (1.5 mL) was stirred at 120° C. for 12 hrs under N2 atmosphere. The mixture was diluted with DCM (30 mL), filtered and washed with DCM/MeOH=10/1 (20 mL). The filtrate was concentrated and purified by prep-TLC to give compound 85-b (63 mg). LCMS (M+H)+: 694.
To a solution of ethyl 2-(4-benzyloxy-5-methyl-2-propyl-pyrazol-3-yl)-4-[3-[(2,4-dimethoxyphenyl)methylcarbamoyl]-6-methyl-imidazo[1,5-a]pyrazin-1-yl]oxazole-5-carboxylate (85-b, 63 mg, 0.09 mmol) in methanol (1 mL) was added 1 N LiOH aq. (1.0 mL). The mixture was stirred at 20° C. for 12 hrs and diluted with water (20 mL). After pH was adjusted to 5 with FA, and the mixture was extracted with DCM (20 mL×2). The combined organic phase was dried with Na2SO4, filtered and concentrated to give compound 85-c (60 mg) as crude solid, which was used directly in the next step. LCMS (M+H)+: 666.
To a solution of 2-(4-benzyloxy-5-methyl-2-propyl-pyrazol-3-yl)-4-[3-[(2,4-dimethoxyphenyl)methylcarbamoyl]-6-methyl-imidazo[1,5-a]pyrazin-1-yl]oxazole-5-carboxylic acid (85-c, 60 mg, 0.09 mmol) in DMSO (1 mL) was added AcOH (2 mg, 0.04 mmol) and Ag2CO3 (10 mg, 0.04 mmol) at 25° C. The reaction was stirred at 80° C. for 2 hrs. The mixture was cooled and poured into water (30 mL), the aqueous phase was extracted with EtOAc (30 mL×2). The combined organic phase was washed with brine (20 mL×3), dried over Na2SO4 and concentrated to give compound 85-d (50 mg, crude). LCMS (M+H)+: 622.
To a solution of 1-[2-(4-benzyloxy-5-methyl-2-propyl-pyrazol-3-yl)oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (85-d, 50 mg, 0.08 mmol) in HFIP (1.0 mL) was added MsOH (0.2 mL). The mixture was stirred at 50° C. for 0.5 h to give a red solution. The mixture was purified directly via prep-HPLC to give Example 85 (11.8 mg). LCMS (M+H)+: 382, 1H NMR (DMSO-d6, 400 MHz) δ=9.65 (d, J=1.6 Hz, 1H), 9.05 (s, 1H), 8.81 (br s, 1H), 8.63 (s, 1H), 8.07 (s, 1H), 7.87 (s, 1H), 4.46 (t, J=7.2 Hz, 2H), 2.47 (s, 3H), 2.16 (s, 3H), 1.89-1.76 (m, 2H), 0.91 (t, J=7.2 Hz, 3H).
Example 86 was prepared in analogy to the preparation of Example 85 by using compound 49-b instead of compound 31-h in step 2.
Example 86 (1.1 mg), LCMS (M+H)+: 396, 1H NMR (400 MHz, DMSO-d6+D2O) δ=9.60 (s, 1H), 8.99 (s, 1H), 8.64 (s, 1H), 4.51-4.43 (m, 2H), 2.43 (s, 3H), 2.13 (s, 3H), 1.80-1.71 (m, 2H), 1.33-1.24 (m, 2H), 0.86 (t, J=7.2 Hz, 3H).
Example 87 was prepared in analogy to the preparation of Example 53 by using compound 87-b instead of 1-bromo-2-phenylethane in step 1 (reaction condition: CMBP, toluene 80° C. instead of K2CO3, DMF 80° C.).
The compound 87-b was prepared according to the following scheme:
To a solution of methyl 2-[4-(bromomethyl)phenyl]acetate (15.0 g, 62 mmol) in methanol (100 mL) was added morpholine (16.0 g, 185 mmol). The mixture was stirred at 25° C. for 3 hrs and concentrated. The residue was treated with water (30 mL) and extracted with ethyl acetate (3×25 mL). The combined organic layer was washed with sat. NaCl aq. (50 mL), dried over sodium sulfate, filtered and concentrated to afford compound 87-a (16.0 g). LCMS (M+H)+: 250.
To a solution of methyl 2-[4-(morpholinomethyl)phenyl]acetate (87-a, 16.0 g, 64 mmol) in THF (200 mL) was added LiAlH4 (30.8 mL, 77 mmol) at 0° C. The reaction was stirred at 0° C. for 2 hrs and then quenched via addition of water (150 mL) and 3 N NaOH aq. (120 mL). The aluminium salts were filtered through celite and the filtrate was concentrated to afford compound 87-b (14.0 g, crude). LCMS (M+H)+: 222.
Example 87 (23.7 mg), LCMS (M+H)+: 543, 1H NMR (400 MHz, DMSO+D2O) δ=9.52 (s, 1H), 8.98 (s, 1H), 7.08-6.93 (m, 4H), 4.64 (br t, J=6.8 Hz, 2H), 3.45 (br s, 4H), 3.27 (s, 2H), 2.95 (br t, J=7.1 Hz, 2H), 2.43 (s, 3H), 2.19 (br s, 4H), 2.11 (s, 3H).
Example 88 was prepared in analogy to the preparation of Example 58 by using compound 88-a instead of compound 31-h.
The compound 88-a was prepared in analogy to the preparation of compound 46-c by using compound 87-b instead of 4-pyridineethanol in step 3.
Example 88 (8.5 mg), LCMS (M+H)+: 543, 1H NMR (400 MHz, DMSO-d6) δ=9.53 (d, J=1.0 Hz, 1H), 9.16-8.84 (m, 1H), 8.66 (s, 1H), 8.05 (s, 1H), 8.00 (s, 1H), 7.81 (s, 1H), 7.32-7.21 (m, 4H), 4.73 (br t, J=7.4 Hz, 2H), 4.25 (s, 2H), 4.00-3.86 (m, 4H), 3.23-3.12 (m, 4H), 3.10-2.98 (m, 2H), 2.67 (s, 3H), 2.16 (s, 3H).
Example 89 was prepared in analogy to the preparation of Example 58 by using compound 89-a instead of compound 31-h.
The compound 89-a was prepared in analogy to the preparation of compound 46-c by using compound 64-a instead of 4-pyridineethanol in step 3.
Example 89 (4.2 mg), LCMS (M+H)+: 488, 1H NMR (400 MHz, DMSO-d6) δ=9.53 (d, J=1.0 Hz, 1H), 9.04-8.87 (m, 1H), 8.66 (s, 1H), 8.04 (s, 1H), 8.00 (s, 1H), 7.80 (s, 1H), 7.14-7.04 (m, 4H), 4.69 (t, J=7.4 Hz, 2H), 4.28 (s, 2H), 3.20 (s, 3H), 3.06 (t, J=7.4 Hz, 2H), 2.67 (s, 3H), 2.17 (s, 3H).
Example 90 was prepared in analogy to the preparation of Example 53 by using compound 90-a instead of 1-bromo-2-phenylethane in step 1 (reaction condition: CMBP, toluene 80° C. instead of K2CO3, DMF 80° C.).
The compound 90-a was prepared according to the following scheme:
Step 1: Preparation of 2-thiazol-2-ylethanol (compound 90-a) A solution of 2-methylthiazole (5 g, 50.43 mmol) in paraformaldehyde (5 g, 50.43 mmol) was slowly heated to 140° C. and stirred for 3 hrs. The reaction mixture was concentrated and purified via column chromatography to afforded compound 90-a (3 g) as colorless oil. 1H NMR (400 MHz, CHLOROFORM-d) δ=7.71 (d, J=3.4 Hz, 1H), 7.24 (d, J=3.4 Hz, 1H), 4.05 (t, J=5.6 Hz, 2H), 3.25 (t, J=5.6 Hz, 2H).
Example 90 (5.6 mg), LCMS (M+H)+: 451, 1H NMR (400 MHz, DMSO-d6+D2O) δ=9.66 (s, 1H), 9.03 (s, 1H), 7.60 (d, J=3.4 Hz, 1H), 7.44 (d, J=3.4 Hz, 1H), 4.83 (br t, J=6.8 Hz, 2H), 3.49 (br t, J=7.0 Hz, 2H), 2.45 (s, 3H), 2.12 (s, 3H).
The title compound was prepared according to the following scheme:
The compound 91-a was prepared in analogy to the preparation of compound 53-e by using 1-iodopropane instead of 1-bromo-2-phenylethane in step 1.
A mixture of 3-(4-benzyloxy-5-methyl-2-propyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazole (91-a, 155 mg, 0.37 mmol), 8-bromo-N-[(2,4-dimethoxyphenyl)methyl]-3-methyl-pyrrolo[1,2-a]pyrazine-6-carboxamide (3-e, 150 mg, 0.37 mmol), Pd(OAc)2 (25 mg, 0.11 mmol), CuI (141 mg, 0.74 mmol), PCy3HBF4 (54 mg, 0.15 mmol), Cs2CO3 (120 mg, 0.37 mmol) in 1,4-dioxane (3 mL) was stirred at 140° C. for 18 hrs under N2 atmosphere. The mixture was cooled, filtered and washed with DCM/MeOH (20 mL, v/v=10/1). The filtrate was concentrated and purified via prep-TLC to give compound 91-b (100 mg). LCMS (M+H)+: 741.
To a solution of 8-[5-(4-benzyloxy-5-methyl-2-propyl-pyrazol-3-yl)-4-[(4-methoxyphenyl)methyl]-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-3-methyl-pyrrolo[1,2-a]pyrazine-6-carboxamide (91-a, 100 mg, 0.13 mmol) in HFIP (2 mL) was added MsOH (259 mg, 2.7 mmol). The mixture was stirred at 50° C. for 0.5 h to give a red solution. The mixture was purified via prep-HPLC to give Example 91 (46.8 mg). LCMS (M+H)+: 381, 1H NMR (DMSO-d6, 400 MHz) δ=13.86 (s, 1H), 9.65 (s, 1H), 9.36 (s, 1H), 9.22-8.77 (m, 1H), 8.16 (s, 2H), 7.55 (s, 1H), 4.43 (br t, J=6.8 Hz, 2H), 2.47 (s, 3H), 2.15 (s, 3H), 1.84-1.73 (m, 2H), 0.86 (t, J=7.4 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of methyl 4-benzyloxy-3-methyl-1H-pyrazole-5-carboxylate (23-d, 2.00 g, 8.1 mmol) in toluene (10 mL) was added 1-propanol (585 mg, 9.7 mmol) and cyanomethylenetributylphosphorane (2550 mg, 10.5 mmol). The reaction was stirred at 80° C. for 12 hrs. The mixture was purified via column chromatography to afford compound 92-a (2.0 g). LCMS (M+H)+: 289.
To a solution of methyl 4-benzyloxy-5-methyl-2-propyl-pyrazole-3-carboxylate (92-a, 870 mg, 3.0 mmol) in methanol (10 mL) was added LiOH·H2O (9.0 mg, 9.0 mmol) in H2O (5 mL). The reaction was stirred at 25° C. for 2 hrs. The mixture was cooled to 0° C. and adjusted pH=3 with 1 N HCl aq, then EtOAc (100 mL) and water 100 mL) were added. After separation, the aqueous phase was extracted with EtOAc (100 mL×3). The combined organic layer was washed with brine (400 mL), dried over Na2SO4, filtered, and concentrated to give compound 92-b (1.6 g, crude). LCMS (M+H)+: 275.
To a solution of 4-benzyloxy-5-methyl-2-propyl-pyrazole-3-carboxylic acid (92-b, 1.5 g, 5.47 mmol) in DCM (30 mL) was added DIPEA (2.1 g, 16.4 mmol) and isobutyl chloroformate (1.5 g, 10.9 mmol). The reaction was stirred at 20° C. for 2 hrs and concentrated to give compound 92-c (2.0 g). LCMS (M+H)+: 375.
To a solution of 1-amino-1-methyl-thiourea (860 mg, 8.15 mmol) in THF (40 mL), was added isobutoxycarbonyl 4-benzyloxy-5-methyl-2-propyl-pyrazole-3-carboxylate (92-c, 1.5 g, 4.07 mmol), then the reaction was stirred at 50° C. for 12 hrs. The mixture was poured into water (200 mL) and extracted with EtOAc (3×200 mL). The combined organic layers was washed with sat. NaHCO3 aq. (300 mL) and brine (300 mL), then dried over anhydrous Na2SO4. The organic layer was concentrated to afford crude compound 92-d (2.0 g). LCMS (M+H)+: 362.
To a solution of 1-[(4-benzyloxy-5-methyl-2-propyl-pyrazole-3-carbonyl)amino]-1-methyl-thiourea (92-d, 2.00 g, 5.5 mmol) in water (12 mL) was added NaOH (720 mg, 18.0 mmol). The reaction was heated to 100° C. and stirred at for 6 hrs. The mixture was poured into water (300 ml) and extracted with EtOAc (3×300 mL). The combined organic layer was washed with sat. NaHCO3 aq. (300 mL) and brine (200 mL), then dried over anhydrous Na2SO4. The filtered organic layer was concentrated to afford compound 92-e (807 mg). LCMS (M+H)+: 344.
To a solution of 5-(4-benzyloxy-5-methyl-2-propyl-pyrazol-3-yl)-2-methyl-1,2,4-triazole-3-thiol (92-e, 800 mg, 2.3 mmol) in acetic acid (6 mL) was added H2O2 (132 mg, 11.6 mmol) slowly at 0° C. The reaction was warmed to 20° C. and stirred for 1 h. The mixture was poured into sat. Na2SO3 aq. (80 mL) and stirred at 0° C. for 30 mins. Then it was adjusted pH to 7-8 with NaHCO3. The mixture was extracted with ethyl acetate (3×50 mL). The combined organic layer was washed with brine (100 mL), dried over Na2SO4, filtered and concentrated. The residue was purified via column chromatography to afford compound 92-f (480 mg). LCMS (M+H)+: 312.
To a solution of 3-(4-benzyloxy-5-methyl-2-propyl-pyrazol-3-yl)-1-methyl-1,2,4-triazole (92-f, 100 mg, 0.32 mmol) in Toluene (0.5 mL) was added 1-bromo-N-[(2,4-dimethoxyphenyl)methyl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (Intermediate A1, 130 mg, 0.32 mmol), catacxiuma (34 mg, 0.1 mmol), Pd(OAc)2 (22 mg, 0.1 mmol), PivOH (20 mg, 0.19 mmol) and K2CO3 (133 mg, 0.96 mmol). The mixture was stirred at 120° C. for 12 hrs and then it was filtered and washed with DCM/MeOH (20/1, 100 ml). The filtrate was concentrated and purified via Pre-TLC to afford compound 92-g (80 mg). LCMS (M+H)+: 636.
To a solution of 1-[5-(4-benzyloxy-5-methyl-2-propyl-pyrazol-3-yl)-2-methyl-1,2,4-triazol-3-yl]-N-[(2,4-dimethoxyphenyl)methyl]-6-methyl-imidazo[1,5-a]pyrazine-3-carboxamide (92-g, 75 mg, 0.12 mmol) in HFIP (1 mL) was added MsOH (226 mg, 2.36 mmol). The mixture was stirred at 50° C. for 1 h. The reaction mixture was filtered and the filtrate was purified via Prep-HPLC to afford Example 92 (22.7 mg) as a yellow solid. LCMS (M+H)+: 396, 1H NMR (400 MHz, DMSO-d6) δ=9.71 (s, 1H), 9.15 (s, 1H), 8.37 (s, 1H), 7.97 (s, 1H), 7.91 (d, J=1.4 Hz, 1H), 4.45 (s, 3H), 4.36 (t, J=7.2 Hz, 2H), 2.51-2.50 (m, 3H), 2.12 (s, 3H), 1.91-1.70 (m, 2H), 0.85 (t, J=7.4 Hz, 3H).
The title compound was prepared according to the following scheme:
To a solution of 4-benzyloxy-5-bromo-3-methyl-1-propyl-pyrazole (31-h, 500 mg, 1.6 mmol) in THF (3 mL) was added n-BuLi (650 μL, 1.6 mmol) dropwise at −63° C. The reaction was stirred at the temperature for 10 mins and ZnCl2 (3.2 mL, 1.6 mmol) was added. Keep stirring at the temperature for 30 min, the reaction was warmed to r.t. and stirred for 1 h. The zincate solution (c=0.23 M) was used in the next step. A mixture of ethyl 2-bromooxazole-5-carboxylate (320 mg, 1.5 mmol) and XPhos-Pd-G3 (137 mg, 0.16 mmol) in 1,4-dioxane (6 mL) was degassed for 5 mins. The above zincate solution was added at 20° C. and the reaction was heated to 80° C. for 90 mins. The reaction was quenched by poured into ice water containing 2.5 mL 1 N HCl. The mixture was extracted with EtOAc (20 mL×3). The combined organic phase was concentrated and purified via flash silica gel chromatography to afford compound 93-a (230 mg). LCMS (M+H)+: 370.
To a solution of ethyl 2-(4-benzyloxy-5-methyl-2-propyl-pyrazol-3-yl)oxazole-5-carboxylate (93-a, 230 mg, 0.62 mmol) in Toluene (3 mL) was added 8-bromo-N-[(2,4-dimethoxyphenyl)methyl]-7-fluoro-3-methyl-pyrrolo[1,2-a]pyrazine-6-carboxamide (4-g, 263 mg, 0.62 mmol), potassium pivalate (131 mg, 0.93 mmol), PivOH (31 mg, 0.31 mmol) and cataCXium-A-Pd-G3 (181 mg, 0.25 mmol). The reaction was stirred at 120° C. for 18 hrs under N2 atmosphere. The mixture was filtered, the wet-cake was washed by DCM/MeOH (V/V=20:1, 30 mL). The filtrate was concentrated and purified via Prep-TLC to give a crude product. Then it was purified again via reversed-phase to afford compound 93-b (40 mg). LCMS (M+H)+: 711.
To a solution of ethyl 2-(4-benzyloxy-5-methyl-2-propyl-pyrazol-3-yl)-4-[6-[(2,4-dimethoxyphenyl)methylcarbamoyl]-7-fluoro-3-methyl-pyrrolo[1,2-a]pyrazin-8-yl]oxazole-5-carboxylate (93-b, 40 mg, 0.06 mmol) in Methanol (4 mL) and water (2 mL) was added LiOH·H2O (12 mg, 0.28 mmol). The reaction was stirred at 20° C. for 12 hrs. The mixture was acidified with HCl (6 N) to pH=3˜4 and extracted with DCM (10 mL×3). The organic layer was washed with brine, dried over Na2SO4 and concentrated to afford crude compound 93-c (33 mg). LCMS (M+H)+: 683.
To a solution of 2-(4-benzyloxy-5-methyl-2-propyl-pyrazol-3-yl)-4-[6-[(2,4-dimethoxyphenyl)methylcarbamoyl]-7-fluoro-3-methyl-pyrrolo[1,2-a]pyrazin-8-yl]oxazole-5-carboxylic acid (93-c, 33 mg, 0.05 mmol) in DMSO (1 mL) was added AcOH (1.2 mg, 0.02 mmol) and Ag2CO3 (5.3 mg, 0.02 mmol) at 20° C. The reaction was stirred at 80° C. for 2 hrs. The mixture was poured into water (10 mL), and extracted with EtOAc (10 mL×2). The combined organic phase was washed with brine (20 mL×3), dried over Na2SO4, filtered and concentrated to afford crude compound 93-d (25 mg). LCMS (M+H)+: 639.
To a solution of 8-[2-(4-benzyloxy-5-methyl-2-propyl-pyrazol-3-yl)oxazol-4-yl]-N-[(2,4-dimethoxyphenyl)methyl]-7-fluoro-3-methyl-pyrrolo[1,2-a]pyrazine-6-carboxamide (93-d, 25 mg, 0.04 mmol) in HFIP (1 mL) was added MsOH (113 mg, 1.17 mmol). The reaction was stirred at 50° C. for 2 hrs to give a red solution. The mixture was filtered and the filtrate was purified via Prep-HPLC to afford Example 93 (13.2 mg). LCMS (M+H)+: 399. 1H NMR (400 MHz, DMSO-d6) δ=9.56 (d, J=1.2 Hz, 1H), 9.14 (s, 1H), 8.83 (br s, 1H), 8.57 (s, 1H), 7.91-7.69 (m, 1H), 7.52-7.30 (m, 1H), 4.44 (t, J=7.2 Hz, 2H), 2.45 (s, 3H), 2.14 (s, 3H), 1.87-1.74 (m, 2H), 0.90 (t, J=7.2 Hz, 3H).
Example 94 was prepared in analogy to the preparation of Example 93 by using compound 3-e instead of compound 4-g in step 2.
Example 94 (43.8 mg), LCMS (M+H)+: 381. 1H NMR (DMSO-d6, 400 MHz) δ=9.51 (d, J=1.2 Hz, 1H), 9.31 (s, 1H), 8.75-8.62 (s, 2H), 8.22-8.05 (m, 1H), 7.99 (s, 1H), 7.54-7.51 (m, 1H), 4.44 (t, J=7.2 Hz, 2H), 2.46 (s, 3H), 2.14 (s, 3H), 1.84-1.74 (m, 2H), 0.88 (t, J=7.2 Hz, 3H).
This is the cellular reporter assay to evaluate compounds' agonism to interferon regulatory factor (IRF) pathway in THP1-Dual™ cells (InvivoGen, cat number thpd-nfis). THP1-Dual™ cells were derived from the human THP-1 monocyte cell line by stable integration lucia luciferase gene, a new secreted luciferase reporter gene, under the control of an ISG54 (interferon-stimulated gene) minimal promoter in conjunction with five interferon (IFN)-stimulated response elements. As a result, THP1-Dual™ cells allow the study of the IRF pathway, by assessing the activity of Lucia luciferase. Lucia luciferase protein is readily measurable in the cell culture supernatant when using QUANTI-Luc™ (InvivoGen, cat: rep-qlcg-500).
After 20-24 hrs compound treatment in THP1-Dual cells, the compounds' agonism to IRF pathway and cell toxicity were tested by measuring luminescence and OD 450 on an Envision plate reader.
On the day of experiment, 30 μL of test medium (RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 1000 heat-inactivated fetal bovine serum) was dispensed in a white 384-well plate (Grenier, cat: 781098) containing the serially diluted compounds (final 1% DMSO) transferred by the Echo550 machine. Then 30 μL of cell suspension (˜33,000 cells, 1.1×106 cells/mL) per well was added immediately by thermo multidrop combi dispenser for incubation 20-24 h at 37° C., 500 CO2. At the end of the incubation, 10 μL of cellular supernatant was transferred to proxiplate 384-plus plate for IRF detection, and then 10 μL of QUANTI-Luc™ Gold solution was added to the plate that proceeded with the measurement immediately. To detect cell viability, 30 μL of the CCK-8 working solution (Dojindo Molecular Technologies, cat: CK04-20) was added to the cell plate, which was Incubated for 2 hrs in the incubator to measure the absorbance at 450 nm using Envision.
2. hPBMC Cytokine Release Assay (Agonist_hPBMC_IFN-O_EC50)
This is the cytokines concentration detection with Luminex assay for human PBMC supernatant. Human peripheral blood mononuclear cells (PBMC) are treated by compounds with appropriate time. Multiple cytokines are released in the supernatant in response to the compound stimulation. Luminex assay can measure the cytokines concentration in supernatant and the compounds activity on human PBMC can be evaluated.
Frozen PBMC was thawed and cultured in PRMI1640 (GIBCO 11875-093) whole medium with 1000 Heat inactivated-FBS (HI—FBS), 1% NEAA, 10% sodium pyruvate and 10% Penicillin-Streptomycin in CO2 incubator overnight. Cells were centrifuged at 400×g for 5 minutes at 4° C. and the pellet was re-suspended with complete RPMI1640 medium to concentration of 1×106 cells/ml in cell culture flask T75 (Thermo Fisher Scientific). Compounds were serially diluted with DMSO and distributed to 96-well flat bottom cell plate (Corning 3599) with Echo555. 90 μL/well PBMC cells were seeded into the plates with Multidrop (Thermo Fisher Scientific). After 24 hours of incubation in CO2 incubator, cells were centrifuged at 400×g for 5 minutes at room temperature, 80 μL supernatant each well was collected for cytokines release test with luminex assay of Human custom ProcartaPlex 5-plex kit (Thermo Fisher Scientific PPX-05). 10 μL supernatant or standards were added to a mixture of color-coded beads, 5 μL Biotinylated detection antibodies specific to the analytes of interest were added and formed an antibody-antigen sandwich. 10 μL Phycoerythrin (PE)-conjugated streptavidin is added to the plate and then read signal of each well on the Luminex 200™. The raw data was analyzed by Millipore Analyst Software and cytokine concentrations were generated accordingly. The EC50 of corresponding cytokine such as interferon-β (IFN-β) was calculated by the 4-parameter sigmoidal concentration response model fitting.
This is the single dose pharmacokinetics (PK) study in female C57/BL6 mice. The purpose of this study was to determine the pharmacokinetics of selected compounds following a single intravenous bolus injection in female C57BL/6J mice. Briefly, 3 female C57BL/6J mice (available from Shanghai Lingchang Biotechnology Co., Ltd) were treated with a single dose of compound intravenously at 1 mg/kg (IV). Blood samples were collected at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 7 h and 24 h post-dose. Blood samples were placed on wet ice until centrifugation to obtain plasma samples. The compound concentration in plasma samples was determined using LC-MS/MS method. The pharmacokinetic parameters were calculated by non-compartmental analysis.
Reference compounds AC01 and AE01 were prepared according to WO 2022/195462 and tested in the assays described above. The results are shown in Table 4 below.
| Number | Date | Country | Kind |
|---|---|---|---|
| PCT/CN2022/093600 | May 2022 | WO | international |
| PCT/CN2022/124648 | Oct 2022 | WO | international |
| PCT/CN2023/080862 | Mar 2023 | WO | international |
| PCT/CN2023/087630 | Apr 2023 | WO | international |
This application is a continuation of International Application No.PCT/EP2023/063042, filed May 16, 2023, which claims the benefit of priority to PCT/CN2023/087630, filed April 11, which 2023, which claims the benefit of priority to PCT/CN2023/080862, filed Mar. 10, 2023, which claims the benefit of priority to PCT/CN2022/124648, filed Oct. 11, 2022, which claims the benefit of priority to PCT/CN2022/093600, filed May 18, 2022 each of which is incorporated herein by reference in its entirety.
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/EP2023/063042 | May 2023 | WO |
| Child | 18947305 | US |
