Derivatives of fgfr inhibitors

Abstract

The present disclosure relates to derivatives of fibroblast growth factor receptor (FGFR) inhibitors (e.g., hydroxyl, keto, glucuronide, sulfonate, and deuterated derivatives), including methods of making the same and intermediates in the preparation thereof, which are useful in the treatment of FGFR-mediated diseases, such as cancer.

Description

Technical Field

[0001] This application relates to derivatives of fibroblast growth factor receptor (FGFR) inhibitors (e.g., hydroxyl, ketone, glucuronic acid, sulfonic acid, and deuterated derivatives), including methods of preparation thereof and intermediates thereof, said substances being used to treat FGFR-mediated diseases such as cancer. Background Technology

[0002] Fibroblast growth factor receptors (FGFRs) are receptor tyrosine kinases that bind to fibroblast growth factor (FGF) ligands. Four FGFR proteins (FGFR1-4) exist, which can bind ligands and participate in the regulation of many physiological processes, including tissue development, angiogenesis, wound healing, and metabolic regulation. Upon ligand binding, the receptor undergoes dimerization and phosphorylation, thereby stimulating protein kinase activity and recruiting numerous intracellular docking proteins. These interactions promote the activation of a series of intracellular signaling pathways, including Ras-MAPK, AKT-PI3K, and phospholipase C, which are important for cell growth, proliferation, and survival (reviewed by Eswarakumar et al. in Cytokine & Growth Factor Reviews, 2005).

[0003] Aberrant activation of this pathway, through overexpression of FGF ligands or FGFR, or through activating mutations in FGFR, can lead to tumor development, progression, and resistance to conventional cancer therapies. In human cancers, genetic alterations leading to ligand-independent receptor activation have been described, including gene amplification, chromosomal translocation, and somatic mutations. Genetic alterations can include mutations, fusions, rearrangements (e.g., translocation, deletion, inversion), and gene amplification. Large-scale DNA sequencing of thousands of tumor samples has shown that components of the FGFR pathway are among the most frequently mutated in human cancers. Many of these activating mutations are identical to germline mutations leading to skeletal dysplasia syndromes. Mechanisms leading to aberrant ligand-dependent signaling in human diseases include FGF overexpression and alterations in FGFR splicing, resulting in a more mixed ligand-binding capacity of the receptor (reviewed in Knights and Cook Pharmacology & Therapeutics, 2010; Turner and Grose, Nature Reviews Cancer, 2010). Therefore, the development of inhibitors targeting FGFR could be used for the clinical treatment of diseases with elevated FGF or FGFR activity.

[0004] Cancer types involving FGF / FGFR include, but are not limited to: carcinomas (e.g., bladder cancer, breast cancer, neck cancer, colorectal cancer, endometrial cancer, gastric cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, ovarian cancer, prostate cancer); hematopoietic malignancies (e.g., multiple myeloma, chronic lymphocytic lymphoma, adult T-cell leukemia, acute myeloid leukemia, non-Hodgkin lymphoma, myeloproliferative neoplasms, and Waldenstrom's macroglubulinemia); and other neoplasms (e.g., glioblastoma, melanoma, and rhabdomyosarcoma). In addition to its role in carcinogenic neoplasms, FGFR activation is also involved in skeletal and chondrocyte disorders, including but not limited to achondroplasia and craniosynostosis. Specifically, the FGFR4-FGF19 signaling axis is involved in the pathogenesis of many cancers, including hepatocellular carcinoma (Heinzle et al., Cur. Pharm. Des. 2014, 20:2881). Ectopic expression of FGF19 in transgenic mice has been shown to lead to tumor formation in the liver, and neutralization of FGF19 with antibodies has been found to inhibit tumor growth in mice. Furthermore, overexpression of FGFR4 has been observed in various tumor types, including hepatocellular carcinoma, colorectal cancer, breast cancer, pancreatic cancer, prostate cancer, lung cancer, and thyroid cancer. Additionally, activating mutations in FGFR4 have been reported in rhabdomyosarcoma (Taylor et al., JCI 2009, 119:3395).

[0005] Inhibitors of FGFR are currently under development for cancer treatment. For example, US Publication Nos. 2012 / 0165305; 2014-0045814; 2013-0338134; 2014 / 0171405; 2014 / 0315902; 2016 / 0115164; 2016 / 0244448; 2016 / 0244449; and 2016-0244450 report the molecule 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-8-(morpholin-4-ylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one and other small molecule inhibitors of FGFR.

[0006] Therefore, there is a continuous need for new or improved FGFR inhibitors to develop new and more effective drugs for the treatment of cancer and other diseases. The compounds (including derivatives of these compounds), compositions, and methods described herein are aimed at these needs and other purposes. Summary of the Invention

[0007] This disclosure provides, in particular, derivatives of compound 1:

[0008]

[0009] Or its pharmaceutically acceptable salt.

[0010] This disclosure also relates to pharmaceutical compositions comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier.

[0011] This disclosure also relates to a method for inhibiting FGFR enzymes, the method comprising contacting the enzyme with a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0012] This disclosure also relates to a method for treating a disease associated with abnormal activity or expression of the FGFR enzyme, the method comprising administering a compound of the disclosure or a pharmaceutically acceptable salt thereof to a patient in need.

[0013] This disclosure also relates to the use of the compounds disclosed herein for the treatment of diseases associated with abnormal activity or expression of FGFR enzymes.

[0014] This disclosure also relates to a method for treating a patient in need of a condition mediated by an FGFR enzyme or a mutant thereof, the method comprising the step of administering to the patient a compound of the present disclosure or a pharmaceutically acceptable composition thereof.

[0015] This disclosure also relates to a method for treating a patient in need of a condition mediated by an FGFR enzyme or a mutant thereof, the method comprising the step of administering to the patient a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof, in combination with another therapy or therapeutic agent as described herein.

[0016] This disclosure also relates to the use of the compounds of this disclosure in the preparation of pharmaceutical agents for use in therapy. Attached Figure Description

[0017] Figure 1 A graph showing the percentage of average cumulative radiation dose recovered in urine and feces at specified intervals after a single oral dose of 13-mg (250 μCi) of compound 1 [14C] was administered to healthy male subjects.

[0018] Figure 2 A graph showing the mean radioactivity (nM equivalent) in blood or plasma and the amount of compound 1 (nM) in plasma after a single oral dose of approximately 13 mg [14c] compound 1 was administered to a healthy male volunteer in a fasting state.

[0019] Figure 3 The mass spectrum of the human circulating metabolite of compound 1 is shown.

[0020] Figure 4 The mass spectrum of the metabolite of compound 1 isolated from urine is shown.

[0021] Figure 5 The mass spectrum of the metabolite of compound 1 isolated from feces is shown. Detailed Implementation

[0022] This disclosure particularly relates to compounds that are derivatives of 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-8-(morpholin-4-ylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (compound 1), and methods and intermediates for preparing said derivatives. The structure of compound 1 is shown below:

[0023]

[0024] Compound 1 is described in U.S. Patent No. 9,611,267, the entire contents of which are incorporated herein by reference.

[0025] In some embodiments, the compound is a metabolite of compound 1. In some embodiments, the compound is an active metabolite that can modulate the activity of one or more FGFR proteins and can be used, for example, to treat diseases associated with FGFR expression or activity. In some embodiments, the levels of the metabolite compounds described herein are measured and analyzed to assist practitioners in adjusting the dosage level of compound 1.

[0026] Therefore, this disclosure also provides a compound of formula I:

[0027]

[0028] Or its pharmaceutically acceptable salt, wherein:

[0029] One or more OCH3 groups may be optionally replaced by OH groups, wherein the OH groups may be optionally replaced by OX groups.

[0030] One CH group may be arbitrarily replaced by a CX group.

[0031] One CH2 group can be arbitrarily replaced by a C=O group.

[0032] One or more CH2 groups of the morpholine ring are optionally replaced by C(OH)H groups, and the NH groups are optionally replaced by NX groups.

[0033] Where X is a group selected from the following:

[0034]

[0035] The prerequisite is that the compound has one or more groups selected from OH, OX, CX, C=O and NX.

[0036] The present invention also provides a compound of formula II:

[0037]

[0038] Or its pharmaceutically acceptable salt, wherein:

[0039] One CH group may be arbitrarily replaced by a CX group.

[0040] One CH2 group can be arbitrarily replaced by a C=O group.

[0041] The NH group can be optionally replaced by an NX group, and

[0042] The OH group can be arbitrarily replaced by an OX group;

[0043] Where X is a group selected from the following:

[0044]

[0045] In some embodiments, the present invention also provides a compound of formula II:

[0046]

[0047] Or a pharmaceutically acceptable salt thereof, wherein one of the CH groups is replaced by a CX group, the NH group by an NX group, or the OH group by an OX group; wherein X is selected from the following groups:

[0048]

[0049] In some embodiments, one or more OCH3 groups are optionally replaced by OH groups, wherein the OH groups are optionally replaced by OX groups. In some embodiments, the OH groups are optionally replaced by OX groups. In some embodiments, a CH group is optionally replaced by a CX group. In some embodiments, a CH2 group is optionally replaced by a C=O group. In some embodiments, the NH group is optionally replaced by an NX group. In some embodiments, one or more CH2 groups of the morpholine ring are each optionally replaced by a C(OH)H group.

[0050] In some embodiments, one or more OCH3 groups are replaced with OH groups, wherein the OH groups are replaced with OX groups. In some embodiments, one or more OCH3 groups are replaced with OH groups. In some embodiments, the OH groups are replaced with OX groups. In some embodiments, a CH group is replaced with a CX group. In some embodiments, a CH2 group is replaced with a C=O group. In some embodiments, an NH group is replaced with an NX group. In some embodiments, one or more CH2 groups of the morpholine ring are each replaced with a C(OH)H group.

[0051] In some implementation schemes, X is In some implementation schemes, X is

[0052] This invention provides a compound, said compound being 3-(2,6-difluoro-3-hydroxy-5-methoxyphenyl)-1-ethyl-8-(morpholinylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (compound 2) or a pharmaceutically acceptable salt thereof. The structure of compound 2 is shown below:

[0053]

[0054] The present invention also provides a compound, said compound being 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-4-hydroxy-8-(morpholinylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (compound 3) or a pharmaceutically acceptable salt thereof. The structure of compound 3 is shown below:

[0055]

[0056] The present invention also provides a compound, said compound being 3-(2,6-difluoro-3,5-dihydroxyphenyl)-1-ethyl-8-(morpholinylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (compound 4) or a pharmaceutically acceptable salt thereof. The structure of compound 4 is shown below:

[0057]

[0058] In some implementation schemes, the compound is selected from:

[0059] 3-(2,6-difluoro-3-hydroxy-5-methoxyphenyl)-1-ethyl-8-(morpholinylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one; and

[0060] 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-4-hydroxy-8-(morpholinylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one,

[0061] Or its pharmaceutically acceptable salt.

[0062] In some embodiments, the compound is a compound of formula I:

[0063]

[0064] Or a pharmaceutically acceptable salt thereof, wherein one CH2 group of the morpholine ring is replaced by a (C=O) group.

[0065] In some embodiments, the compound is a compound of formula II:

[0066]

[0067] Or a pharmaceutically acceptable salt thereof, wherein one of the CH groups is replaced by a CX group, the NH group by an NX group, or the OH group by an OX group; wherein X is one of the following groups:

[0068]

[0069] In some embodiments, the compound is a compound of formula II:

[0070]

[0071] Or a pharmaceutically acceptable salt thereof, wherein one of the CH groups is replaced by a CX group, the NH group by an NX group, or the OH group by an OX group; wherein X is one of the following groups:

[0072]

[0073] In some embodiments, the compound is a compound of formula I:

[0074]

[0075] Or a pharmaceutically acceptable salt thereof, wherein the two CH2 groups of the morpholine ring are each replaced by a C(OH)H group.

[0076] In some embodiments, the compound is a compound of formula I:

[0077]

[0078] Or a pharmaceutically acceptable salt thereof, wherein each OCH3 group is replaced by an OH group; and one CH2 group of the morpholine ring is replaced by a (C=O) group.

[0079] In some embodiments, the compound is a compound of formula II:

[0080]

[0081] Or a pharmaceutically acceptable salt thereof, wherein one CH2 group of the morpholine ring is replaced by a (C=O) group.

[0082] This article also provides a compound, said compound being 3-(2,6-difluoro-3-hydroxy-5-methoxyphenyl)-1-ethyl-2-oxo-2,3,4,7-tetrahydro-1H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidine-8-carboxylic acid (compound 5) or a pharmaceutically acceptable salt thereof.

[0083] The structure of compound 5 is shown below:

[0084]

[0085] This document also provides a compound, said compound being 3-(2,6-difluoro-3,5-bis(methoxy-d3)phenyl)-1-ethyl-8-(morpholinylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (compound 6) or a pharmaceutically acceptable salt thereof. The structure of compound 6 is shown below:

[0086]

[0087] Each of the above implementation schemes assumes adherence to the rules of appropriate pricing.

[0088] Certain compounds described herein are metabolites of 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-8-(morpholin-4-ylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (compound 1). The metabolites were isolated from human blood / serum, urine, and fecal samples collected from pharmacokinetic and toxicokinetic studies of compound 1. The metabolites of this invention may be FGFR inhibitors and may possess advantageous properties (PK, PD, toxicity, etc.) compared to the parent compound (compound 1). For example, a metabolite of this disclosure (e.g., compound 2) may be a better substrate for Pgp transport than compound 1. Therefore, a metabolite of this disclosure (e.g., compound 2) may be a more suitable candidate for intravenous administration or hepatic artery infusion for the treatment of the diseases and conditions disclosed herein (e.g., cholangiocarcinoma).

[0089] In some embodiments, the compounds of the present invention are substantially isolated. "Substantially isolated" means that the compound is at least partially or substantially separated from the environment in which it is formed or detected. Partial isolation may include, for example, compositions rich in the compounds of the present invention. Substantially isolated may include compositions containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% of metabolites by weight.

[0090] This document also provides a composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof, said composition containing at least about 50% by weight of said compound or salt. In some embodiments, the composition contains at least about 60% by weight of the compound or salt. In some embodiments, the composition contains at least about 70% by weight of the compound or salt. In some embodiments, the composition contains at least about 80% by weight of the compound or salt. In some embodiments, the composition contains at least about 90% by weight of the compound or salt.

[0091] Method for preparing compound 2

[0092] This application also provides a method for preparing compound 2, wherein the method is adaptable for scale-up.

[0093] This article provides a method for preparing compound 2 or a salt thereof having the following formula:

[0094]

[0095] The method includes deprotecting compound F4 or a salt thereof having the following formula:

[0096]

[0097] Where P 1 It is an amino protecting group. In some embodiments, P 1 It is a sulfonamide group. In some embodiments, P 1 It is -SO2N(CH3)2.

[0098] In some embodiments, deprotection of compound F4 involves reacting compound F4 with A1, wherein A1 is an acid. In some embodiments, A1 is hydrochloric acid. In some embodiments, A1 is an aqueous solution of hydrochloric acid. In some embodiments, deprotection is carried out at a temperature of 70-90°C (e.g., 80°C). In some embodiments, deprotection is carried out in the presence of S1, wherein S1 is a polar aprotic solvent. In some embodiments, S1 is 1,4-dioxane. In some embodiments, deprotection of compound F4 involves using a deprotecting agent in an amount of about 1 to about 50 molar equivalents relative to compound F4, an amount of a deprotecting agent in an amount of about 10 to about 30 molar equivalents relative to compound F4, or an amount of a deprotecting agent in an amount of about 20 molar equivalents relative to compound F4.

[0099] Compound F4 or its salts can be prepared by a method comprising the following steps: making compound F3 having the following formula:

[0100]

[0101] Or its salt reacts with morpholine in the presence of RA1, wherein RA1 is a reducing agent. In some embodiments, RA1 is sodium triacetoxyborohydride. In some embodiments, the reaction of compound F3 with RA1 is carried out in the presence of A2, wherein A2 is an acid. In some embodiments, A2 is an organic acid. In some embodiments, A2 is acetic acid. In some embodiments, the reaction of compound F3 with RA1 is carried out in the presence of S2, wherein S2 is a solvent. In some embodiments, S2 is a polar aprotic solvent. In some embodiments, S2 is methane chloride.

[0102] Compound F3 or its salts can be prepared by a method comprising the following steps: making compound F2 having the following formula:

[0103]

[0104] The reaction of compound F2 or its salt with DMF in the presence of B1, wherein B1 is a base. In some embodiments, B1 is lithium diisopropylamide (“LDA”). In some embodiments, the reaction of compound F2 with DMF in the presence of B1 is carried out in the presence of S3, wherein S3 is a polar aprotic solvent. In some embodiments, S3 is tetrahydrofuran. The reaction of compound F2 with DMF in the presence of B1 can be carried out at a temperature between about -100°C and about -50°C (e.g., about -64°C).

[0105] Compound F2 or its salt can be prepared by a method comprising the following steps: making compound F1 having the following formula:

[0106]

[0107] Or its salts and containing P 1 The reaction of amino protecting agents.

[0108] In some implementations, the amino protecting agent is P 1 -X, where X is a halogen. In some embodiments, the amino protecting agent is Me₂NSO₂Cl. The reaction of compound F1 with the amino protecting agent can be carried out in the presence of B₂, where B₂ is a base. In some embodiments, B₂ is a metal hydroxide base. In some embodiments, B₂ is NaOH.

[0109] The reaction of compound F1 with the amino protecting agent can be carried out in the presence of tetrabutylammonium bisulfate. In some embodiments, the reaction of compound F1 with the amino protecting agent is carried out in the presence of S4, wherein S4 is a polar aprotic solvent. In some embodiments, S4 is tetrahydrofuran. In some embodiments, the reaction of compound F1 with the amino protecting agent is carried out at a temperature of about 0°C to about 50°C (e.g., about 0°C to about 30°C).

[0110] This article provides a method for preparing compound 2 or a salt thereof having the following formula:

[0111]

[0112] The method includes:

[0113] a) Make compound F1 have the following formula:

[0114]

[0115] With P 1 The amino protecting agent reacts to provide compound F2 having the following formula:

[0116]

[0117] or its salt, wherein P 1 It is an amino protecting group;

[0118] b) React compound F2 with DMF in the presence of B1, where B1 is a base, to provide compound F3 having the following formula:

[0119]

[0120] or its salt;

[0121] c) React compound F3 with morpholine in the presence of RA1, where RA1 is a reducing agent, to provide compound F4 having the following formula:

[0122]

[0123] or its salt; and

[0124] d) Deprotect compound F4 to provide compound 2 or its salt.

[0125] Method for preparing compound 6

[0126] This disclosure provides a method for preparing compound 6 having the following formula or a salt thereof:

[0127]

[0128] The method includes deprotecting compound F5 or a salt thereof having the following formula:

[0129]

[0130] Where P 2 It is an amino protecting group. In some embodiments, P 2 For -SO 2 -Phenyl.

[0131] Deprotection of compound F5 may include treating compound F5 with B3, wherein B3 is a base. In some embodiments, B3 is a metal hydroxide base. In some embodiments, B3 is NaOH. In some embodiments, B3 is an aqueous solution of NaOH. In some embodiments, deprotection of compound F5 is carried out in the presence of S5, wherein S5 is 1,4-dioxane. In some embodiments, the reaction of compound F5 with B3 includes using about 1 to about 10 molar equivalents of B3 relative to compound F5, about 2 to about 8 molar equivalents of B3 relative to compound F5, or about 4 molar equivalents of B3 relative to compound F5.

[0132] Compound F5 can be prepared by a method including the following steps: making compound F6 having the following formula:

[0133]

[0134] It reacts with CD3I in the presence of B4, where B4 is a base. In some embodiments, B4 is sodium hydride.

[0135] The reaction of compound F6 with CD3I can be carried out in the presence of S6, wherein S6 is a polar aprotic solvent. In some embodiments, S6 is DMF. The reaction of compound F6 with CD3I may include using about 1 to about 5 molar equivalents of CD3I relative to compound F6, about 1 to about 3 molar equivalents of CD3I relative to compound F6, or about 2 molar equivalents of CD3I relative to compound F6. The reaction of compound F6 with CD3I may include using about 1 to about 10 molar equivalents of B4 relative to compound F6, about 2 to about 8 molar equivalents of B4 relative to compound F6, or about 4 to about 5 molar equivalents of B4 relative to compound F6.

[0136] Compound F6 can be prepared by a method including the following steps: making compound F7 having the following formula:

[0137]

[0138] It reacts with A3, where A3 is a Lewis acid. In some embodiments, A3 is BBr3.

[0139] The reaction of compound F7 with A3 can be carried out in the presence of S7, which is a polar aprotic solvent. In some embodiments, S7 is methyl chloride. The reaction of compound F7 with A3 can be carried out at a temperature of about -100°C to about 30°C (e.g., about -100°C to room temperature). The reaction of compound F7 with A3 may include using about 1 to about 20 molar equivalents of A3 relative to compound F7, about 5 to about 15 molar equivalents of A3 relative to compound F7, or about 8 to about 12 molar equivalents of A3 relative to compound F7.

[0140] This article provides a method for preparing compound 6 or a salt thereof having the following formula:

[0141]

[0142] The method includes:

[0143] a) Make compound F7 have the following formula:

[0144]

[0145] It reacts with A3, where A3 is a Lewis acid, and where P 2 The amino protecting group provides compound F6 having the following formula:

[0146]

[0147] or its salt;

[0148] b) React compound F6 with CD3I in the presence of B4, where B4 is a base, to provide compound F5 having the following formula:

[0149]

[0150] or its salt; and

[0151] c) Deprotect compound F5 to provide compound 6 or its salt.

[0152] The methods described herein can be monitored using any suitable method known in the art. For example, they can be monitored using spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., 1 H or 13 C) Infrared spectroscopy, spectrophotometry (e.g., UV-Vis), or mass spectrometry; or by chromatography, such as high-performance liquid chromatography (HPLC) or thin-layer chromatography, to monitor product formation. The compound obtained by the reaction can be purified by any suitable method known in the art. For example, chromatography (medium pressure), HPLC, or preparative thin-layer chromatography on a suitable adsorbent (e.g., silica gel, alumina, etc.); distillation; sublimation, grinding, or recrystallization. Generally, the purity of a compound is determined by physical methods, such as measuring the melting point (in the solid case), obtaining an NMR spectrum, or performing HPLC separation. A compound can be considered purified if the melting point decreases, if an undesirable signal is reduced in the NMR spectrum, or if irrelevant peaks in the HPLC record are removed. In some embodiments, the compound is substantially purified.

[0153] The preparation of compounds can involve the protection and deprotection of various chemical groups. The need for protection and deprotection, and the selection of appropriate protecting groups, can be readily determined by those skilled in the art. The chemical role of protecting groups can be found, for example, in Wuts and Greene, Greene's Protective Groups in Organic Synthesis, 4th Edition, John Wiley & Sons: New York, 2006, which are incorporated herein by reference in their entirety. As used herein, "amino protecting group" means any protecting group used to protect amines. Exemplary amino protecting groups include, but are not limited to, phenylsulfonyl, benzyloxycarbonyl (Cbz), 2,2,2-trichloroethoxycarbonyl (Troc), 2-(trimethylsilyl)ethoxycarbonyl (Teoc), 2-(4-trifluoromethylphenylsulfonyl)ethoxycarbonyl (Tsc), tert-butoxycarbonyl (BOC), 1-adamantoxycarbonyl (Adoc), 2-adamantylcarbonyl (2-Adoc), 2,4-dimethylpentyl-3-yloxycarbonyl (... Doc), cyclohexyloxycarbonyl (Hoc), 1,1-dimethyl-2,2,2-trichloroethoxycarbonyl (TcBOC), vinyl, 2-chloroethyl, 2-phenylsulfonylethyl, allyl, benzyl, 2-nitrobenzyl, 4-nitrobenzyl, diphenyl-4-pyridylmethyl, N',N'-dimethylhydrazyl, methoxymethyl, tert-butoxymethyl (Bum), benzyloxymethyl (BOM) or 2-tetrahydropyranyl (THP), tri(C 1-4 Alkyl)silyl (e.g., tris(isopropyl)silyl), 1,1-diethoxymethyl or N-tervapotranoxymethyl (POM).

[0154] The reactions described herein can be carried out at a suitable temperature, which can be readily determined by a skilled technician. The reaction temperature will depend, for example, the melting and boiling points of the reagents and solvents (if present); the thermodynamics of the reaction (e.g., a violently exothermic reaction may require a low temperature); and the kinetics of the reaction (e.g., a high activation energy barrier may require a high temperature).

[0155] In some embodiments, concentration of a solution as described herein refers to reducing the volume of the solution by evaporating the solvent, heating the solution, subjecting the solution to reduced pressure, or any combination thereof.

[0156] The reactions described herein can be carried out in suitable solvents, which can be readily selected by those skilled in organic synthesis. Suitable solvents are generally unreactive with the starting materials (reactants), intermediates, or products at temperatures in which the reaction is carried out (e.g., temperatures in the range of the solvent's freezing to boiling temperature). A given reaction can be carried out in one solvent or a mixture of more than one solvent. Depending on the specific reaction step, a suitable solvent can be selected for that particular reaction step. In some embodiments, such as when at least one of the reagents is a liquid or a gas, the reaction can be carried out in the absence of a solvent.

[0157] Suitable solvents may include halogenated solvents such as carbon tetrachloride, bromodichloromethane, dibromochloromethane, bromoform, chloroform, bromochloromethane, dibromomethane, butyl chloride, dichloromethane (chloromethane), tetrachloroethylene, trichloroethylene, 1,1,1-trichloroethane, 1,1,2-trichloroethane, 1,1-dichloroethane, 2-chloropropane, α,α,α-trifluorotoluene, 1,2-dichloroethane, 1,2-dibromoethane, hexafluorobenzene, 1,2,4-trichlorobenzene, 1,2-dichlorobenzene, chlorobenzene, fluorobenzene, and mixtures thereof.

[0158] Suitable ether solvents include: dimethoxymethane, tetrahydrofuran, 1,3-dioxane, 1,4-dioxane, furan, tetrahydrofuran (THF), diethyl ether, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, diethylene glycol dimethyl ether (diethylene glycol dimethyl ether), diethylene glycol diethyl ether, triethylene glycol dimethyl ether, benzyl ether, tert-butyl methyl ether, and mixtures thereof.

[0159] Suitable proton solvents may include, for example, but not limited to, water, methanol, ethanol, 2-nitroethanol, 2-fluoroethanol, 2,2,2-trifluoroethanol, ethylene glycol, 1-propanol, 2-propanol, 2-methoxyethanol, 1-butanol, 2-butanol, isobutanol, tert-butanol, 2-ethoxyethanol, diethylene glycol, 1-pentanol, 2-pentanol or 3-pentanol, neopentanol, tert-pentanol, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, cyclohexanol, benzyl alcohol, phenol or glycerol.

[0160] Suitable aprotic solvents may include, for example, but not limited to, N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA), 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU), 1,3-dimethyl-2-imidazolidineone (DMI), N-methylpyrrolidone (NMP), formamide, N-methylacetamide, N-methylformamide, acetonitrile, dimethyl sulfoxide, propionitrile, ethyl formate, methyl acetate, hexachloroacetone, acetone, ethyl methyl ketone, ethyl acetate, sulfolane, N,N-dimethylpropionamide, tetramethylurea, nitromethane, nitrobenzene, or hexamethylphosphoramide.

[0161] Suitable hydrocarbon solvents include benzene, cyclohexane, pentane, hexane, toluene, cycloheptane, methylcyclohexane, heptane, ethylbenzene, m-xylene, o-xylene or p-xylene, octane, dihydroindene, nonane or naphthalene.

[0162] The reactions described herein can be carried out in air or in an inert atmosphere. Typically, air-sensitive synthesis techniques familiar to skilled technicians can be used to carry out reactions involving reagents or products that are generally reactive with air.

[0163] As used herein, “ambient temperature” and “room temperature” are as understood in the art and generally refer to a temperature approximately equal to the temperature of the room where the reaction takes place (e.g., the reaction temperature), such as about 20°C to about 30°C. As used herein, the term “high temperature” is as understood in the art and generally refers to a temperature above room temperature (e.g., the reaction temperature), such as above 30°C.

[0164] This article provides information on compound F2:

[0165]

[0166] or its salt, wherein P 1 It is an amino protecting group.

[0167] In some embodiments, compound F2 has the following structure:

[0168]

[0169] Or its salt.

[0170] This article provides information on compound F3:

[0171]

[0172] or its salt, wherein P 1 It is an amino protecting group.

[0173] In some embodiments, compound F3 has the following structure:

[0174]

[0175] Or its salt.

[0176] This article provides information on compound F4:

[0177]

[0178] or its salt, wherein P 1 It is an amino protecting group.

[0179] In some embodiments, compound F4 has the following structure:

[0180]

[0181] Or its salt.

[0182] This article provides information on compound F5:

[0183]

[0184] or its salt, wherein P 2 It is an amino protecting group.

[0185] In some embodiments, compound F5 has the following structure:

[0186]

[0187] Or its salt.

[0188] This article provides information on compound F6:

[0189]

[0190] or its salt, wherein P 2 It is an amino protecting group.

[0191] In some embodiments, compound F6 has the following structure:

[0192]

[0193] Or its salt.

[0194] The compounds disclosed herein also include tautomer forms. Tautomer forms result from the exchange of single bonds with adjacent double bonds and the accompanying proton migration. Tautomer forms include proton-mutant tautomers, which are isoprotonated states having the same empirical formula and total charge. Exemplary proton-mutant tautomers include keto-enol pairs, amide-imine pairs, lactam-lactamimide pairs, enamine-imide pairs, and cyclic forms, wherein the proton can occupy two or more positions in the heterocyclic system, such as 1H-imidazole and 3H-imidazole; 1H-1,2,4-triazole, 2H-1,2,4-triazole, and 4H-1,2,4-triazole; 1H-isoindole and 2H-isoindole; and 1H-pyrazole and 2H-pyrazole. Tautomer forms can be in equilibrium or spatially locked into one form by appropriate substitution.

[0195] The compounds disclosed herein also include all isotopes of the atoms appearing in the intermediates or final compounds. Isotopes include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. One or more constituent atoms of the compounds disclosed herein may be substituted or replaced by isotopes of atoms of natural or non-natural abundance. In some embodiments, the compound includes at least one deuterium atom. For example, one or more hydrogen atoms in the compounds disclosed herein may be substituted or replaced by deuterium. In some embodiments, the compound includes two or more deuterium atoms. In some embodiments, the compound includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 deuterium atoms. Synthetic methods that incorporate isotopes into organic compounds are known in the art (Deuterium Labeling in Organic Chemistry, Alan F. Thomas (New York, NY, Appleton-Century-Crofts, 1971; The Renaissance of H / D Exchange, Jens Atzrodt, Volker Derdau, Thorsten Fey, and Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765; The Organic Chemistry of Isotopic Labelling, James R. Hanson, Royal Society of Chemistry, 2011). Isotope-labeled compounds can be used in various studies, such as NMR spectroscopy, metabolic experiments, and / or analysis.

[0196] Replacing with heavy isotopes (such as deuterium) can provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dose requirements, and is therefore preferred in some cases. (A. Kerekes et al. J. Med. Chem. 2011, 54, 201-210; R. Xu et al. J. Label Compd. Radiopharm. 2015, 58, 308-312).

[0197] As used herein, the term "compound" is intended to include all stereoisomers, geometric isomers, tautomers, and isotopes of the structures shown. The term is also intended to refer to the compounds of this disclosure, regardless of how they are prepared, such as synthetically, by biological processes (e.g., metabolism or enzymatic conversion), or combinations thereof.

[0198] How to use

[0199] The compounds described herein can inhibit the activity of FGFR enzymes. For example, the compounds of this disclosure can be used to inhibit the activity of the enzyme in cells, individuals, or patients to which inhibition of FGFR enzymes is required.

[0200] As FGFR inhibitors, the compounds disclosed herein can be used to treat various diseases associated with the abnormal expression or activity of FGFR enzymes or FGFR ligands. Compounds that inhibit FGFR will be used to provide means of preventing growth or inducing apoptosis in tumors, particularly by inhibiting angiogenesis. Therefore, the compounds disclosed herein are expected to prove useful in the treatment or prevention of proliferative conditions such as cancer. In particular, tumors with activated mutants of receptor tyrosine kinases or with upregulated receptor tyrosine kinases may be particularly sensitive to said inhibitors.

[0201] In some embodiments, this disclosure provides a method for treating FGFR-mediated conditions in patients of need, the method comprising the step of administering a compound of this disclosure or a pharmaceutically acceptable composition thereof to the patient.

[0202] For example, the compounds disclosed herein can be used to treat cancer. Exemplary cancers include bladder cancer, breast cancer (e.g., hormone R positive, triple negative), cervical cancer, colorectal cancer, small bowel cancer, colon cancer, rectal cancer, anal cancer, endometrial cancer, gastric cancer (e.g., gastrointestinal stromal tumor), head and neck cancer (e.g., laryngeal cancer, hypopharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, lip cancer, oral cancer, and squamous head and neck cancer), kidney cancer (e.g., renal cell carcinoma, urothelial carcinoma, sarcoma, Wilms tumor), liver cancer (e.g., hepatocellular carcinoma, cholangiocarcinoma, hepatic angiosarcoma, hepatoblastoma), lung cancer (e.g., adenocarcinoma, small cell lung cancer, non-small cell lung cancer, small cell and non-small cell carcinoma, bronchial cancer, bronchial adenoma, pleural pulmonary germ cell tumor), ovarian cancer, prostate cancer, testicular cancer, uterine cancer, vulvar cancer, esophageal cancer, gallbladder cancer, pancreatic cancer (e.g., exocrine pancreatic cancer), gastric cancer, thyroid cancer, parathyroid cancer, neuroendocrine cancer (e.g., pheochromocytoma, Merkel cell carcinoma). Cancers include neuroendocrine cancers, skin cancers (e.g., squamous cell carcinoma, Kaposi sarcoma, Merck cell carcinoma), and brain cancers (e.g., astrocytoma, medulloblastoma, ependymoma, neuroectodermal tumor, pineal tumor).

[0203] Other exemplary cancers include hematopoietic malignancies such as leukemia or lymphoma, multiple myeloma, chronic lymphocytic lymphoma, adult T-cell leukemia, B-cell lymphoma, cutaneous T-cell lymphoma, acute myeloid leukemia, Hodgkin's or non-Hodgkin's lymphoma, myeloproliferative neoplasms (e.g., 8p11 myeloproliferative syndrome, polycythemia vera, spontaneous thrombocytosis, and primary myelofibrosis), myelodysplastic syndromes, chronic eosinophilic leukemia, Waldenström's macroglobulinemia, pilocellular lymphoma, chronic myeloid lymphoma, acute lymphoblastic lymphoma, AIDS-related lymphoma, and Burkitt's lymphoma.

[0204] In some embodiments, this article provides a method for treating myeloma / lymphoma in patients of need. In some embodiments, the myeloma / lymphoma is 8p11 myeloproliferative syndrome. As used herein, the term "8p11 myeloproliferative syndrome" (EMS) refers to myeloma / lymphoma associated with eosinophilia and FGFR1 abnormalities or myeloma / lymphoma (MLN) with FGFR1 rearrangements. 8p11 myeloproliferative syndrome is reviewed in Jackson, Courtney C. et al., Human Pathology, 2010, 41, 461-476. Identifying features of EMS are the presence of translocations involving the FGFR1 gene located at the chromosomal locus 8p11, and at least 10 additional translocations and 1 insertion have been identified in the EMS, each disrupting FGFR1 and forming novel fusion genes with various partner bodies. See Jackson, Courtney C. et al., Human Pathology, 2010, 41, 461-476.

[0205] In some embodiments, myeloma / lymphoma is characterized by FGF / FGFR gene alterations. Gene alterations may include mutations, fusions, rearrangements (e.g., translocations, deletions, inversions), and gene amplifications. In some embodiments, myeloma / lymphoma exhibits FGFR1 fusions. FGFR1 fusions may be translocations, intermediate deletions, or chromosomal inversions. In some embodiments, FGFR1 fusions are FGFR1 translocations. In some embodiments, myeloma / lymphoma exhibits 8p11 translocations. In some embodiments, 8p11 translocations are associated with FGFR1 activation. In some embodiments, myeloma / lymphoma exhibits FGF / FGFR alterations that are not FGFR1 translocations. In some embodiments, the patient has failed at least one prior treatment for myeloma / lymphoma (e.g., 8p11 myeloproliferative syndrome). In some embodiments, the prior treatment was surgery or radiation therapy. In some embodiments, the patient has a history of hepatitis. In some embodiments, the hepatitis is chronic hepatitis B or hepatitis C. In some embodiments, the patient does not have a history of hepatitis.

[0206] In some embodiments, this document provides a method for treating cancer, the method comprising administering a therapeutically effective amount of the disclosed compound to a patient in need. In some embodiments, the cancer is selected from bladder cancer, breast cancer, cervical cancer, small bowel cancer, colorectal cancer, endometrial cancer, gastric cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, ovarian cancer, prostate cancer, testicular cancer, uterine cancer, vulvar cancer, esophageal cancer, gallbladder cancer, pancreatic cancer, thyroid cancer, skin cancer, brain cancer, leukemia, multiple myeloma, chronic lymphocytic lymphoma, adult T-cell leukemia, B-cell lymphoma, acute myeloid leukemia, Hodgkin's or non-Hodgkin's lymphoma, Waldenström's macroglobulinemia, myeloproliferative neoplasm, chronic myeloid lymphoma, acute lymphoblastic lymphoma, T-cell lymphoblastic lymphoma, pilocellular lymphoma, Burkitt's lymphoma, glioblastoma, melanoma, rhabdomyosarcoma, lymphosarcoma, and osteosarcoma.

[0207] In some implementations, the cancer is bladder cancer (e.g., urothelial carcinoma, squamous cell carcinoma, adenocarcinoma).

[0208] In some embodiments, hepatocellular carcinoma is cholangiocarcinoma (e.g., intrahepatic, hilar, or perihepatic, distal extrahepatic). As used herein, cholangiocarcinoma is synonymous with cholangiocarcinoma tumor or cholangiocarcinoma. In some embodiments, cholangiocarcinoma is advanced or metastatic cholangiocarcinoma. In some embodiments, cholangiocarcinoma is surgically unresectable cholangiocarcinoma. In some embodiments, cholangiocarcinoma is intrahepatic cholangiocarcinoma. In some embodiments, cholangiocarcinoma is extrahepatic cholangiocarcinoma. In some embodiments, cholangiocarcinoma exhibits an FGFR2 tyrosine kinase fusion, which defines a molecular subtype as described in Arai, Yasuhito et al., Hepatology, 2014, 59, 1427-1434. In some embodiments, cholangiocarcinoma is characterized by tumors with altered FGF / FGFR genes. In some embodiments, the tumor exhibits an FGFR2 fusion. FGFR2 fusion can be a translocation, intermediate deletion, or chromosomal inversion. In some embodiments, FGFR2 fusion is an FGFR2 translocation. FGFR2 translocation can be selected from, but is not limited to, the following groups: FGFR2-BICC1, FGFR2-AHCYL1, FGFR2-MACF1, and FGFR2 intron 17 rearrangement. In some embodiments, the tumor exhibits FGF / FGFR alterations that are not due to FGFR2 translocation. In some embodiments, the cholangiocarcinoma does not exhibit FGF / FGFR gene alterations.

[0209] Other cancers that can be treated with the compounds disclosed herein include ocular tumors, glioblastomas, melanomas, rhabdomyosarcomas, lymphosarcomas, leiomyosarcomas, urothelial carcinomas (e.g., ureteral cancer, urethral cancer, bladder cancer, urachal cancer), and osteosarcomas.

[0210] The compounds disclosed herein can also be used to inhibit tumor metastasis.

[0211] In some implementations, compounds of this disclosure as described herein can be used to treat Alzheimer's disease, HIV, or tuberculosis.

[0212] In addition to carcinogenic lesions, the compounds disclosed herein can be used to treat bone and chondrocyte disorders, including but not limited to achondroplasia, reduced chondrogenesis, dwarfism, lethal dysplasia (TD) (clinical forms TD I and TD II), Apert syndrome, Crouzon syndrome, Jackson-Weiss syndrome, Beare-Stevenson cutisgyrate syndrome, Pfeiffer syndrome, and craniosynostosis syndrome.

[0213] The compounds described herein can also be used to treat fibrotic diseases, such as those in which fibrosis is a characteristic symptom or condition. Exemplary fibrotic diseases include cirrhosis, glomerulonephritis, pulmonary fibrosis, systemic fibrosis, rheumatoid arthritis, and wound healing.

[0214] In some implementations, the compounds provided herein can be used to treat hypophosphatemia conditions such as X-linked hypophosphatemic rickets, autosomal recessive hypophosphatemic rickets, autosomal dominant hypophosphatemic rickets, or tumor-induced osteomalacia.

[0215] In some embodiments, this document provides a method for increasing patient survival or progression-free survival, the method comprising administering a compound provided herein to a patient. In some embodiments, the patient has cancer. In some embodiments, the patient has the disease or condition described herein. In some embodiments, the patient has cholangiocarcinoma. In some embodiments, this document provides a method for increasing the survival or progression-free survival of a patient with cholangiocarcinoma characterized by FGFR2 fusion, the method comprising administering a compound provided herein to a patient. As used herein, progression-free survival refers to the length of time a patient lives with the disease without it worsening during and after treatment for a solid tumor. Progression-free survival may refer to the length of time from the first administration of the compound to earlier death or disease progression. Disease progression can be determined using RECIST version 1.1 (Standard for Evaluation of Response to Solid Tumors), as evaluated by an independent centralized radiological review committee. In some embodiments, the progression-free survival induced by the application of the compound is greater than about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 8 months, about 9 months, about 12 months, about 16 months, or about 24 months. In some embodiments, the progression-free survival induced by the application of the compound is at least about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 8 months, about 9 months, or about 12 months; and less than about 24 months, about 16 months, about 12 months, about 9 months, about 8 months, about 6 months, about 5 months, about 4 months, about 3 months, or about 2 months. In some implementations, the application of the compound increases progression-free survival by at least about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 8 months, about 9 months, or about 12 months; and by less than about 24 months, about 16 months, about 12 months, about 9 months, about 8 months, about 6 months, about 5 months, about 4 months, about 3 months, or about 2 months.

[0216] As used herein, the term "cell" means a cell in vitro, outside the body, or in vivo. In some embodiments, the cell in vitro may be a portion of a tissue sample excised from an organism such as a mammal. In some embodiments, the cell in vitro may be a cell in a cell culture. In some embodiments, the cell in vivo may be a cell that is alive in an organism such as a mammal.

[0217] As used herein, the term “contact” means bringing together the indicated portions in an in vitro or in vivo system. For example, “contacting” an FGFR enzyme with a compound described herein (e.g., compound 1) includes administering the compound described herein to an individual or patient (such as a human) who has FGFR, and, for example, introducing the compound described herein (e.g., compound 1) into a sample containing cells or a purified formulation containing an FGFR enzyme.

[0218] As used herein, the interchangeable terms “individual” or “patient” refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, and most preferably humans.

[0219] As used herein, the phrase "therapeutic effective amount" refers to the amount of an active compound or agent that elicits a biological or medical response sought by an investigator, veterinarian, physician, or other clinician in a tissue, system, animal, individual, or human, such as in any of the solid forms disclosed herein or any salts thereof. The appropriate "effective" amount in any individual case can be determined using techniques known to those skilled in the art.

[0220] The phrase “pharmaceutically acceptable” is used herein to refer to compounds, materials, compositions, and / or dosage forms that are suitable for contact with human and animal tissues to the extent reasonably medically permissible without excessive toxicity, irritation, allergic reactions, immunogenicity, or other problems or complications, and that meet a reasonable benefit / risk ratio.

[0221] As used herein, the phrase "pharmaceuticalally acceptable carrier or excipient" refers to a pharmaceutically acceptable material, composition, or medium, such as a liquid or solid filler, diluent, solvent, or encapsulating material. Excipients or carriers are generally safe, non-toxic, and neither biologically nor otherwise undesirable, and include excipients or carriers acceptable for veterinary use as well as for human pharmaceutical use. In one embodiment, each component is defined herein as "pharmaceuticalally acceptable." See, for example, Remington: The Science and Practice of Pharmacy, 21st edition; Lippincott Williams & Wilkins: Philadelphia, Pa., 2005; Handbook of Pharmaceutical Excipients, 6th edition; Rowe et al., eds.; The Pharmaceutical Press and the American Pharmaceutical Association: 2009; Handbook of Pharmaceutical Additives, 3rd edition; Ash and Ash, eds.; Gower Publishing Company: 2007; Pharmaceutical Preformulation and Formulation, 2nd edition; Gibson, ed.; CRC Press LLC: Boca Raton, Fla., 2009.

[0222] As used herein, the term “treating / treatment” refers to suppressing a disease; for example, suppressing the disease, illness, or condition of an individual who experiences or exhibits lesions or symptoms of the disease, disorder, or symptom (i.e., preventing further development of the lesions and / or symptoms), or improving a disease; for example, improving the disease, illness, or symptom of an individual who experiences or exhibits lesions or symptoms of the disease, disorder, or symptom (even if the lesions and / or symptoms are reversed), such as reducing the severity of the disease.

[0223] It should be understood that certain features described in the context of individual embodiments in this disclosure for clarity may also be provided in combination in a single embodiment (while the embodiments are intended to be combined, as if written in multiple independent forms). Conversely, various features described in the context of individual embodiments in this disclosure for simplicity may also be provided individually or in any suitable sub-combination.

[0224] Combination therapy

[0225] One or more additional agents or treatments (such as antiviral agents, chemotherapy agents or other anticancer agents, immune enhancers, immunosuppressants, radiation, antitumor and antiviral vaccines, cytokine therapy (e.g., IL2, GM-CSF, etc.) and / or tyrosine kinase inhibitors) may be combined with the compounds described herein to treat diseases, conditions, or disorders related to FGFR, or as described herein. The agents may be combined with the compounds of the present invention in a single dosage form, or the agents may be administered simultaneously or sequentially as separate dosage forms.

[0226] Compounds as described herein can be combined with one or more other kinase inhibitors for the treatment of diseases affected by multiple signaling pathways, such as cancer. For example, combinations may include one or more inhibitors of the following kinases for cancer treatment: Akt1, Akt2, Akt3, TGF-βR, Pim, PKA, PKG, PKC, CaM-kinase, phosphorylase kinase, MEKK, ERK, MAPK, mTOR, EGFR, HER2, HER3, HER4, INS-R, IGF-1R, IR-R, PDGFαR, PDGFβR, CSFIR, KIT, FLK-II, KDR / FL K-1, FLK-4, flt-1, FGFR1, FGFR2, FGFR3, FGFR4, c-Met, Ron, Sea, TRKA, TRKB, TRKC, FLT3, VEGFR / Flt2, Flt4, EphA1, EphA2, EphA3, EphB2, EphB4, Tie2, Src, Fyn, Lck, Fgr, Btk, Fak, SYK, FRK, JAK, ABL, ALK, and B-Raf. Additionally, the solid form of FGFR inhibitors as described herein can be combined with inhibitors of kinases associated with the PIK3 / Akt / mTOR signaling pathway, such as PI3K, Akt (including Akt1, Akt2, and Akt3), and mTOR kinase.

[0227] In some embodiments, compounds as described herein may be used in combination with one or more inhibitors of enzymes or protein receptors such as HPK1, SBLB, TUT4, A2A / A2B, CD47, CDK2, STING, ALK2, LIN28, ADAR1, MAT2a, RIOK1, HDAC8, WDR5, SMARCA2, and DCLK1. Exemplary diseases and conditions include cancer, infections, inflammation, and neurodegenerative diseases.

[0228] In some implementations, the compounds described herein may be used in combination with therapeutic agents that target epigenetic regulators. Examples of epigenetic regulators include bromodomain inhibitors, histone lysine methyltransferases, histone arginine methyltransferases, histone demethylases, histone deacetylases, histone acetylases, and DNA methyltransferases. Histone deacetylase inhibitors include, for example, vorinostat.

[0229] For the treatment of cancer and other proliferative diseases, compounds as described herein can be used in combination with targeted therapies, including JAK kinase inhibitors (ruxolitinib, additional JAK1 / 2 and JAK1 selective inhibitors, baricitinib, or INCB39110), Pim kinase inhibitors (e.g., LGH447, INCB053914, and SGI-1776), PI3 kinase inhibitors (including PI3K-δ selective and broad-spectrum PI3K inhibitors, such as INCB50465 and INCB54707), PI3K-γ inhibitors (such as PI3K-γ selective inhibitors), MEK inhibitors, CSF1R inhibitors (e.g., PLX3397 and LY3022855), TAM receptor tyrosine kinase inhibitors (Tyro-3, Axl, and Mer; e.g., INCB81776), angiogenesis inhibitors, interleukin receptor inhibitors, cyclin-dependent kinase inhibitors, BRAF inhibitors, and mTOR inhibitors. Inhibitors, proteasome inhibitors (bortezomib, carfilzomib), HDAC inhibitors (panobinostat, vorinostat), DNA methyltransferase inhibitors, dexamethasone, inhibitors of bromine and additional terminal family members (e.g., bromine domain inhibitors or BET inhibitors, such as OTX015, CPI-0610, INCB54329 or INCB57643), LSD1 inhibitors (e.g., GSK2979552, INCB59872 and INCB60003), arginase inhibitors (e.g., INCB1158), indoleamine 2,3-dioxygenase inhibitors (e.g., epacadostat, NLG919 or BMS-986205), PARP inhibitors (e.g., olaparib or rucaparib), and BTK inhibitors, such as ibrutinib. In addition, for the treatment of cancer and other proliferative diseases, the compounds described herein can be used in combination with targeted therapies such as c-MET inhibitors (e.g., capmatinib), anti-CD19 antibodies (e.g., tafasitamab), ALK2 inhibitors (e.g., INCB00928); or combinations thereof.

[0230] For the treatment of cancer and other proliferative diseases, compounds as described herein can be used in combination with chemotherapeutic agents, nuclear receptor agonists or antagonists, or other antiproliferative agents. Compounds described herein can also be used in combination with medical therapies such as surgery or radiation therapy, for example gamma radiation, neutron beam radiation therapy, electron beam radiation therapy, proton therapy, brachytherapy, and systemic radioisotopes.

[0231] Examples of suitable chemotherapy agents include any of the following: abarrelix, abiraterone, afatinib, aflibercept, aldesleukin, alemtuzumab, alitretinoin, allopurinol, hexamethylmelamine, amidox, amsacrine, anastrozole, and aph. Idicolon), arsenic trioxide, asparaginase, axitinib, azacitidine, bevacizumab, bexarotene, baricitinib, bendamustine, bicalutamide, bleomycin, bortezombi / bortezomib, brivanib, buparlisib, and intravenous busulfan. Intravenous, oral busulfan, calusterone, camptosar, capecitabine, carboplatin, carmustine, cediranib, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, crizotinib, cyclophosphamide, cytarabine, dacarbazine, dacomitinib, dactinomycin, dalteparin sodium, dasatinib, daunorubicin, decitabine, degarelix, denileukin, denileukin-diftitox, deoxycoformycin, dexrazoxane, didox, docetaxel, doxorubicin, droloxafine, dromostanolonepropionate, eculizumab, enzalutamide, epidophyllotoxin, epirubicin, epothilone, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, fluorouracil, fludarabine, fluorouracil, flutamide, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate, histrelin acetate Acetate), ibritumomab tiuxetan, idarubicin, idelalisib, ifosfamide, imatinib mesylate, interferon alpha-2a, irinotecan, lapatinib ditosylate, lenalidomide, letrozole, leuprolide acetate, levamisole, lonafarnib, lomustine, meclorethamine, medroxyprogesterone acetate, melphalan, mercaptopurine, methotrexate, methoxsalen, mithramycin, mitomycin CC) Mitotane, Mitoxantrone, Nandrolone Phenpropionate, Navelbene, Necitumumab, Nelarabine, Neratinib, Nilotinib, Nilutamide, Niraparib, Nofetumomab, Oserelin, Oxaliplatin, Paclitaxel, Pamidronate, Panitumumab, Pabbisat, Pazopanib, Pegaspargase, Pegfilgrastim, PemetrexedDisodium, pentostatin, pilaralisib, pipobroman, plicamycin, ponatinib, porfimer, prednisone, procarbazine, quinacrine, ranibizumab, rasburicase, regorafenib, reloxafine, revlimid, rituximab, rucapranib, ruxotinib, sorafenib, streptozocin, sunitinib, sunitinib maleate, tamoxifen, tegafur, temozolomide, and tinib. Teniposide, testosterone, tezacitabine, thalidomide, thioguanine, thiotepa, tipifarnib, topotecan, toremifene, tositumomab, trastuzumab, retinoic acid, triapine, trimidox, triptorelin, uracil mustard, valrubicin, vandetanib, vinblastine, vincristine, vindesine, vinorelbine, vorinostat, veliparib, talazoparib, and zoledronate.

[0232] In some embodiments, the compounds described herein may be used in combination with immune checkpoint inhibitors. Exemplary immune checkpoint inhibitors include inhibitors against immune checkpoint molecules such as CD27, CD28, CD40, CD122, CD96, CD73, CD47, OX40, GITR, CSF1R, JAK, PI3Kδ, PI3Kγ, TAM, arginase, CD137 (also known as 4-1BB), ICOS, A2AR, B7-H3, B7-H4, BTLA, CTLA-4, LAG3 (e.g., INCAGN2385), TIM3 (e.g., INCB2390), VISTA, PD-1, PD-L1, and PD-L2. In some embodiments, the immune checkpoint molecule is selected from stimulating checkpoint molecules such as CD27, CD28, CD40, ICOS, OX40 (e.g., INCAGN1949), GITR (e.g., INCAGN1876), and CD137. In some embodiments, the immune checkpoint molecule is an inhibitory checkpoint molecule selected from the following: A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, TIM3, and VISTA. In some embodiments, the compounds provided herein may be used in combination with one or more agents selected from the following: KIR inhibitors, TIGIT inhibitors, LAIR1 inhibitors, CD160 inhibitors, 2B4 inhibitors, and TGFRβ inhibitors.

[0233] In some embodiments, the inhibitor of the immune checkpoint molecule is a small molecule PD-L1 inhibitor. In some embodiments, the small molecule PD-L1 inhibitors described in the PD-L1 assays described in U.S. Patent Publications US 20170107216, US 20170145025, US 20170174671, US 20170174679, US 20170320875, US 20170342060, US 20170362253, and US 20180016260 have IC50 values ​​of less than 1 μM, less than 100 nM, less than 10 nM, or less than 1 nM, each of which is incorporated herein by reference in its entirety for all purposes.

[0234] In some embodiments, the inhibitor of the immune checkpoint molecule is a PD-1 inhibitor, such as an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 monoclonal antibody is MGA012 (retifanlimab), nivolumab, pembrolizumab (also known as MK-3475), pidilizumab, SHR-1210, PDR001, ipilimumab, or AMP-224. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab or pembrolizumab. In some embodiments, the anti-PD1 antibody is pembrolizumab. In some embodiments, the anti-PD1 antibody is nivolumab. In some embodiments, the anti-PD-1 monoclonal antibody is MGA012 (retifanlimab). In some embodiments, the anti-PD1 antibody is SHR-1210. One or more other anticancer agents include antibody therapeutics, such as 4-1BB (e.g., urelumab, utomilumab).

[0235] In some embodiments, the compounds disclosed herein may be used in combination with INCB086550.

[0236] In some embodiments, the inhibitor of the immune checkpoint molecule is a PD-L1 inhibitor, such as an anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-L1 monoclonal antibody is BMS-935559, MEDI4736, MPDL3280A (also known as RG7446), or MSB0010718C. In some embodiments, the anti-PD-L1 monoclonal antibody is MPDL3280A or MEDI4736.

[0237] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of CTLA-4, such as an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab, tremelimumab, AAGEN1884, or CP-675,206.

[0238] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of LAG3, such as an anti-LAG3 antibody. In some embodiments, the anti-LAG3 antibody is BMS-986016, LAG525, or INCAGN2385.

[0239] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of TIM3, such as an anti-TIM3 antibody. In some embodiments, the anti-TIM3 antibody is INCAGN2390, MBG453, or TSR-022.

[0240] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of GITR, such as an anti-GITR antibody. In some embodiments, the anti-GITR antibody is TRX518, MK-4166, INCAGN1876, MK-1248, AMG228, BMS-986156, GWN323, or MEDI1873.

[0241] In some embodiments, the inhibitor of the immune checkpoint molecule is an agonist of OX40, such as an OX40 agonist or an OX40L fusion protein. In some embodiments, the anti-OX40 antibody is MEDI0562, MOXR-0916, PF-04518600, GSK3174998, or BMS-986178. In some embodiments, the OX40L fusion protein is MEDI6383.

[0242] In some embodiments, the inhibitor of the immune checkpoint molecule is a CD20 inhibitor, such as an anti-CD20 antibody. In some embodiments, the anti-CD20 antibody is obinutuzumab or rituximab.

[0243] The compounds disclosed herein can be used in combination with bispecific antibodies. In some embodiments, one of the domains of the bispecific antibody targets a PD-1, PD-L1, CTLA-4, GITR, OX40, TIM3, LAG3, CD137, ICOS, CD3, or TGFβ receptor.

[0244] In some embodiments, the compounds of this disclosure may be used in combination with one or more metabolic enzyme inhibitors. In some embodiments, the metabolic enzyme inhibitor is an inhibitor of IDO1, TDO, or arginase. Examples of IDO1 inhibitors include icardolstat, NLG919, BMS-986205, PF-06840003, IOM2983, RG-70099, and LY338196.

[0245] In some embodiments, the compounds described herein may be used in combination with one or more agents for treating diseases such as cancer. In some embodiments, the agents are alkylating agents, proteasome inhibitors, corticosteroids, or immunomodulators. Examples of alkylating agents include cyclophosphamide (CY), melphalan (MEL), and bendamustine. In some embodiments, the proteasome inhibitor is carfilzomib. In some embodiments, the corticosteroid is dexamethasone (DEX). In some embodiments, the immunomodulator is lenalidomide (LEN) or pomalidomide (POM).

[0246] Suitable antiviral agents intended for use in combination with the compounds disclosed herein may include nucleoside and nucleotide reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), protease inhibitors, and other antiviral agents.

[0247] Examples of suitable NRTIs include zidovudine (AZT); didanosine (ddl); zalcitabine (ddC); stavudine (d4T); lamivudine (3TC); abacavir (1592U89); and adefovir dipivoxil. dipivoxil (bis(POM)-PMEA); lobucavir (BMS-180194); BCH-10652; emitricitabine ((-)-FTC); β-L-FD4 (also known as β-L-D4C and named β-L-2',3'-dideoxy-5-fluorocytidine); DAPD ((-)-β-D-2,6,-diamino-purine dioxane); and lodenosine (FddA). Typical suitable NNRTIs include nevirapine (BI-RG-587); delaviradine (BHAP, U-90152); efavirenz (DMP-266); PNU-142721; AG-1549; MKC-442 (1-(ethoxy-methyl)-5-(1-methylethyl)-6-(phenylmethyl)-(2,4(1H,3H)-pyrimidinidone); and (+)-calanolide A (NSC-675451) and B. Typical suitable protease inhibitors include saquinavir (Ro... 31-8959); ritonavir (ABT-538); indinavir (MK-639); nelfnavir (AG-1343); amprenavir (141W94); lasinavir (BMS-234475); DMP-450; BMS-2322623; ABT-378; and AG-1 549. Other antiviral agents include hydroxyurea, ribavirin, IL-2, IL-12, pentafuside, and Yissum Project No. 11607.

[0248] Suitable agents for use in combination with the compounds described herein to treat cancer include chemotherapy agents, targeted cancer therapies, immunotherapy, or radiotherapy. The compounds described herein, in combination with antihormonal agents, may be effective in treating breast cancer and other tumors. Suitable examples include antiestrogens, including but not limited to tamoxifen and toremifene; aromatase inhibitors, including but not limited to letrozole, anastrozole, and exemestane; adrenocorticosteroids (e.g., prednisone), progesterone (e.g., medroxyprogesterone acetate), and estrogen receptor antagonists (e.g., fulvestrant). Antihormonal agents suitable for treating prostate and other cancers may also be combined with the compounds described herein. These antihormonal agents include antiandrogens, including but not limited to flutamide, bicamid, and nilumet; luteinizing hormone-releasing hormone (LHRH) analogs, including leuprorelin, goserelin, triptorelin, and histaminerelin; LHRH antagonists (e.g., degarelix); androgen receptor blockers (e.g., enzalutamide); and androgen inhibitors (e.g., abiraterone).

[0249] The compounds described herein can be used in combination with or sequentially with other agents targeting membrane receptor kinases, especially in patients who have developed primary or acquired resistance to targeted therapies. These agents include inhibitors or antibodies against EGFR, Her2, VEGFR, c-Met, Ret, IGFR1, or Flt-3, as well as against cancer-associated fusion protein kinases such as Bcr-Abl and EML4-Alk. EGFR inhibitors include gefitinib and erlotinib, and EGFR / Her2 inhibitors include, but are not limited to, dacomitinib, afatinib, lapatinib, and neratinib. EGFR antibodies include, but are not limited to, cetuximab, panitumumab, and nexituzumab. c-Met inhibitors can be used in combination with FGFR inhibitors. These include onartumzumab, tivantnib, and INC-280. Agents targeting Abl (or Bcr-Abl) include imatinib, dasatinib, nilotinib, and ponatinib, while those targeting Alk (or EML4-ALK) include crizotinib.

[0250] Angiogenesis inhibitors can be effective in combination with FGFR inhibitors in some tumors. These substances include antibodies against VEGF or VEGFR, or VEGFR kinase inhibitors. Antibodies against VEGF or other therapeutic proteins include bevacizumab and aflibercept. VEGFR kinase inhibitors and other anti-angiogenic inhibitors include, but are not limited to, sunitinib, sorafenib, axitinib, cediranib, pazopanib, regorafenib, brinib, and vandetanib.

[0251] Activation of intracellular signaling pathways occurs frequently in cancer, and agents targeting components of these pathways have been combined with receptor-targeting agents to enhance efficacy and reduce resistance. Examples of agents that can be combined with the compounds described herein include inhibitors of the PI3K-AKT-mTOR pathway, Raf-MAPK pathway, JAK-STAT pathway, and protein chaperones and cell cycle progression inhibitors.

[0252] Agents targeting PI3 kinase include, but are not limited to, piraprine, edrexate, and bupalixate. mTOR inhibitors such as rapamycin, sirolimus, temsirolimus, and everolimus can be combined with FGFR inhibitors. Other suitable examples include, but are not limited to, vemurafenib and dabrafenib (Raf inhibitors), trametinib, selumetinib, and GDC-0973 (MEK inhibitor). The compounds described herein may also be combined with one or more inhibitors of JAK (e.g., ruxotinib, baricitinib, tofacitinib), Hsp90 (e.g., tanespimycin), cyclin-dependent kinases (e.g., palbociclib), HDAC (e.g., pabisostat), PARP (e.g., olaparib), and proteasomes (e.g., bortezomib, carfilzomib). In some embodiments, the JAK inhibitors are selective for JAK1 compared to JAK2 and JAK3.

[0253] Other agents suitable for use in combination with the compounds described herein include chemotherapy combinations such as platinum-based dual therapies for lung cancer and other solid tumors (cisplatin or carboplatin plus gemcitabine; cisplatin or carboplatin plus docetaxel; cisplatin or carboplatin plus parcitabine; cisplatin or carboplatin plus pemetrexed) or gemcitabine plus parcitabine combined particles.

[0254] Suitable chemotherapeutic agents or other anticancer agents include, for example, alkylating agents (including but not limited to nitrogen mustard, ethyleneimine derivatives, alkyl sulfonates, nitrosoureas, and triazines), such as uracil nitrogen mustard, chlormethine, and cyclophosphamide. TM ), ifosfamide, melphalan, nitrogen mustard chlorpyrifos, piperobromo, triethylene melamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, strepzotocin, dacarbazine, and temozolomide.

[0255] Other agents suitable for use in combination with the compounds described herein include steroids (including 17α-ethinylestradiol), diethylhexethrin, testosterone, prednisone, fluoromethyltestosterone, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, trichlorotriaryl, hydroxyprogesterone, aminoglutethimide, and medroxyprogesterone acetate.

[0256] Other agents suitable for use in combination with the compounds described herein include: dacarbazine (DTIC), optionally with other chemotherapy agents such as carmustine (BCNU) and cisplatin; the "Dartmouth regimen," which consists of DTIC, BCNU, cisplatin, and tamoxifen; a combination of cisplatin, vinblastine, and DTIC; or temozolomide. The compounds described herein may also be combined with immunotherapy agents, including cytokines such as interferon-alpha, interleukin-2, and tumor necrosis factor (TNF).

[0257] Suitable chemotherapy agents or other anticancer agents include, for example, antimetabolites (including but not limited to folic acid antagonists, pyrimidine analogs, purine analogs and adenosine deaminase inhibitors), such as methotrexate, 5-fluorouracil, fluorouracil, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, pentostatin and gemcitabine.

[0258] Suitable chemotherapeutic agents or other anticancer agents also include, for example, certain natural products and their derivatives (such as vinca alkaloids, antitumor antibiotics, enzymes, lymphokines, and epipodophyllotoxin), such as vinca alkaloids, vincristine, vindesine, bleomycin, daunomycin, doxorubicin, epirubicin, idarubicin, ara-C, and palcitabine (TAXOL). TM ), sclerosingmycin, deoxycofromycin, mitomycin-C, L-asparaginase, interferon (especially IFN-α), etoposide, and teniposide.

[0259] Other cytotoxic agents include navitabine, CPT-11, anastrozole, letrozole, capecitabine, raloxafine, cyclophosphamide, ifosfamide, and raloxafine.

[0260] Cytotoxic agents are also suitable, such as epipodophyllotoxin; antitumor enzymes; topoisomerase inhibitors; procarbazine; mitoxantrone; platinum coordination complexes, such as cisplatin and carboplatin; biological response modifiers; growth inhibitors; anti-hormonal therapeutic agents; leucovorin; tegafur; and hematopoietic growth factors.

[0261] One or more other anticancer agents include antibody therapeutics, such as trastuzumab (Herceptin), antibodies against co-stimulatory molecules (such as CTLA-4, 4-1BB), PD-1 and PD-L1 antibodies, or antibodies against cytokines (IL-10, TGF-β, etc.).

[0262] Other anticancer agents include those that block the migration of immune cells, such as antagonists of chemokine receptors including CCR2 and CCR4.

[0263] Other anticancer agents include those that enhance the immune system, such as adjuvants or adoptive T-cell transfer.

[0264] Cancer vaccines include dendritic cells, synthetic peptides, DNA vaccines, and recombinant viruses. In some embodiments, tumor vaccines comprise proteins derived from viruses involved in human cancers, such as human papillomavirus (HPV), hepatitis viruses (HBV and HCV), and Kaposi's herpes sarcoma virus (KHSV). Non-limiting examples of usable tumor vaccines include peptides of melanoma antigens (such as gp100, MAGE antigen, Trp-2, MARTI, and / or tyrosinase peptides) or tumor cells transfected to express the cytokine GM-CSF.

[0265] The compounds disclosed herein can be used in combination with bone marrow transplantation for the treatment of various hematopoietic tumors.

[0266] Methods for the safe and effective administration of most of these chemotherapeutic agents are known to those skilled in the art. Furthermore, their administration is described in standard literature. For example, the administration of many chemotherapeutic agents is described in the Physicians' Desk Reference (PDR, e.g., 1996 edition, Medical Economics Company, Montvale, NJ), the disclosure of which is incorporated herein by reference as if it were described in its entirety.

[0267] As provided throughout, other compounds, inhibitors, agents, etc., may be combined with the compounds of the present invention in single or sequential dosage forms, or they may be administered simultaneously or sequentially as individual dosage forms.

[0268] Pharmaceutical formulations and dosage forms

[0269] In some embodiments, the compounds or pharmaceutical compositions thereof disclosed herein are suitable for oral administration. In some embodiments, the compounds or pharmaceutical compositions thereof disclosed herein are suitable for intravenous administration. In some embodiments, the compounds or pharmaceutical compositions thereof disclosed herein are suitable for arterial administration. In some embodiments, arterial administration is by hepatic artery infusion.

[0270] When used as a pharmaceutical, the compounds described herein may be administered in the form of pharmaceutical compositions, wherein the pharmaceutical composition refers to a combination of one or more compounds as described herein with at least one pharmaceutically acceptable carrier. The pharmaceutical compositions disclosed herein may contain 20% by weight of the compound or a salt thereof. The pharmaceutical compositions disclosed herein may contain 30% by weight of the compound or a salt thereof. The pharmaceutical compositions disclosed herein may contain 40% by weight of the compound or a salt thereof. The pharmaceutical compositions disclosed herein may contain 50% by weight of the compound or a salt thereof. The pharmaceutical compositions disclosed herein may contain 60% by weight of the compound or a salt thereof. The pharmaceutical compositions disclosed herein may contain 70% by weight of the compound or a salt thereof. The pharmaceutical compositions disclosed herein may contain 80% by weight of the compound or a salt thereof. The pharmaceutical compositions disclosed herein may contain 90% by weight of the compound or a salt thereof. The pharmaceutical compositions disclosed herein may contain 95% by weight of the compound or a salt thereof. The pharmaceutical compositions disclosed herein may contain 99% by weight of the compound or a salt thereof.

[0271] These compositions can be prepared in ways well known in pharmaceutical technology and can be administered via a variety of routes, depending on whether they are for local or systemic treatment and the area to be treated. Administration can be topically (including via the eyes and mucous membranes, including intranasal, vaginal, and rectal delivery), pulmonaryly (e.g., by inhalation or blowing of powders or aerosols, including via nebulizers; intratracheal, intranasal, transepidermal, and transdermal), ocularly, orally, or parenterally. Methods of ocular delivery can include topical application (eye drops), subconjunctival, perivitelline, or intravitreal injection, or introduction via a balloon catheter or surgically placed ocular insert into the conjunctival sac. Parenteral administration includes intravenous, intra-arterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion; or intracranial, such as intrathecal or intraventricular administration. Parenteral administration can be in the form of a single bolus dose or, for example, via a continuous infusion pump. Pharmaceutical compositions and formulations for topical application can include dermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional drug carriers; aqueous, powder, or oil-based matrices; thickeners, etc., may be necessary or required.

[0272] This disclosure also includes pharmaceutical compositions comprising a compound of the present disclosure as an active ingredient and one or more pharmaceutically acceptable carriers. In preparing the compositions described herein, the active ingredient is typically mixed with an excipient, which is then diluted or sealed within such a carrier, for example, in the form of capsules, sachets, paper, or other containers. When the excipient acts as a diluent, it can be a solid, semi-solid, or liquid material that serves as a medium, carrier, or substrate for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, capsules, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), creams containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterilely packaged powders.

[0273] When preparing a formulation, the active compound can be ground to provide an appropriate particle size before being combined with other ingredients. If the active compound is substantially insoluble, it can be ground to a particle size of less than 200 mesh. If the active compound is substantially water-soluble, the particle size can be adjusted by grinding to provide a substantially uniform distribution in the formulation, for example, about 40 mesh.

[0274] Examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth gum, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. Formulations may further include: lubricants such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifiers and suspending agents; preservatives such as methyl benzoate and propylparaben; sweeteners; and flavoring agents. The compositions described herein can be formulated to provide a rapid, sustained, or delayed release of the active ingredient upon administration to a patient using procedures known in the art.

[0275] The composition may be formulated as a unit dosage form, each dose containing about 5 to about 100 mg, more typically about 10 to about 30 mg of the active ingredient. The term "unit dosage form" refers to a physically discrete unit suitable for use as a single dose in human subjects and other mammals, each unit containing a predetermined amount of active material calculated to combine with suitable pharmaceutical excipients to produce the desired therapeutic effect.

[0276] Active compounds are effective over a wide dose range and are usually administered at pharmaceutically effective amounts. However, it should be understood that the actual amount of compound administered will generally be determined by the physician based on relevant circumstances, including the condition to be treated, the route of administration chosen, the actual compound administered, the individual patient's age, weight, response, and the severity of the patient's symptoms.

[0277] For the preparation of solid compositions such as tablets, the main active ingredient is mixed with a pharmaceutical excipient to form a solid preformed composition containing a homogeneous mixture of compounds of the present disclosure. When these preformed compositions are referred to as homogeneous, the active ingredient is typically uniformly dispersed in the composition, such that the composition can be easily further divided into equally effective unit dosage forms, such as tablets, pills, and capsules. This solid preformed composition is then further divided into unit dosage forms of the type described above, wherein the unit dosage form contains, for example, 0.1 to about 500 mg of the active ingredient of the present disclosure.

[0278] The tablets or pills of this disclosure can be coated or otherwise formulated to provide dosage forms with the advantage of prolonged action. For example, the tablets or pills may contain an internal dose component and an external dose component, the latter being in the form of a coating on the former. The two components may be separated by an enteric coating layer, which resists disintegration in the stomach and allows the internal component to be properly delivered into the duodenum or to have a delayed release. A variety of materials can be used for such enteric coatings or coatings, including many polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

[0279] The compounds and compositions of this disclosure as described herein may be incorporated herein in liquid forms for oral or injectable administration, including aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, emulsions flavored with edible oils (such as cottonseed oil, sesame oil, coconut oil, or peanut oil), as well as elixirs and similar pharmaceutical carriers.

[0280] Compositions for inhalation or inhalation include solutions and suspensions, as well as powders, in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof. As described above, liquid or solid compositions may contain suitable pharmaceutically acceptable excipients. In some embodiments, the composition is administered via the oral or nasal respiratory route to achieve local or systemic effects. The composition can be nebulized using an inert gas. The nebulized solution can be obtained directly from the nebulizer or the nebulizer can be connected to a face mask or intermittent positive pressure ventilation machine. Solutions, suspensions, or powder compositions can be administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0281] The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of administration (such as prevention or treatment), the patient's condition, the method of administration, etc. In therapeutic applications, an amount sufficient to cure or at least partially suppress the symptoms and complications of the disease may be administered to a patient already suffering from the disease. The effective dose will depend on the disease being treated and will be determined by the attending clinician based on factors such as the severity of the disease, the patient's age, weight, and general condition.

[0282] The compositions administered to patients may be in the form of the pharmaceutical compositions described above. These compositions may be sterilized using conventional sterilization techniques or may be sterile filtered. Aqueous solutions may be packaged as is or lyophilized, with the lyophilized formulation combined with a sterile aqueous carrier prior to administration. The pH of the compound formulation is typically between 3 and 11, more preferably between 5 and 9, and most preferably between 7 and 8. It should be understood that the use of some of the aforementioned excipients, carriers, or stabilizers may cause the formation of pharmaceutical salts.

[0283] The therapeutic dose of the compounds described herein may vary depending on, for example, the specific purpose of treatment, the route of administration, the patient's health and condition, and the prescribing physician's judgment. The proportion or concentration of the compounds described herein in a pharmaceutical composition may vary depending on many factors, including dosage, chemical properties (e.g., hydrophobicity), and route of administration. For example, the compounds described herein may be provided in a physiologically buffered aqueous solution containing about 0.1 to about 10% w / v of the compound for parenteral administration. Some typical dosage ranges are about 1 μg to about 1 g per kilogram of body weight per day. In some embodiments, the dosage range is about 0.01 mg to about 100 mg per kilogram of body weight per day. The dosage may depend on variables such as the type and progression of the disease or condition, the overall health status of the particular patient, the relative biological efficacy of the selected compound, the formulation of the excipients, and the route of administration. The effective dose can be inferred from dose-response curves obtained from in vitro or animal model testing systems.

[0284] The compounds disclosed herein can also be formulated in combination with one or more additional active ingredients, which may include any pharmaceutical agent, such as an antiviral agent, vaccine, antibody, immune enhancer, immunosuppressant, anti-inflammatory agent, etc.

[0285] Example

[0286] The experimental procedures for the compounds used in this invention are provided below. Preparative LC-MS purification of some of the prepared compounds was performed on a Waters mass-directed fractionation system. The basic equipment setup, procedures, and control software for operating these systems have been described in detail in the literature. See, for example, "Two-Pump At Column Dilution Configuration for Preparative LC-MS", K. Blom, J. Combi. Chem., 4, 295 (2002); "Optimizing Preparative LC-MS Configurations and Methods for ParallelSynthesis" Purification", K. Blom, R. Sparks, J. Doughty, G. Everlof, T. Haque, A. Combs, J. Combi. Chem., 5, 670 (2003); and "Preparative LC-MS Purification: Improved Compound Specific Method Optimization", K. Blom, B. Glass, R. Sparks, A. Combs, J. Combi. Chem., 6, 874-883 (2004). Typically, the separated compounds are subjected to analytical liquid chromatography-mass spectrometry (LC-MS) for purity analysis under the following conditions: Instrument: Agilent 1100 series, LC / MSD, Column: Waters Sunfire TM C 18 5 μm, 2.1 x 50 mm, buffer: mobile phase A: 0.025% TFA aqueous solution and mobile phase B: acetonitrile; the gradient of B is 2% to 80% over 3 minutes and the flow rate is 2.0 mL / min.

[0287] Furthermore, at the preparative scale, some of the prepared compounds were separated by reversed-phase high-performance liquid chromatography (RP-HPLC) or rapid chromatography (silica gel) with an MS detector, as indicated in the examples. Typical preparative RP-HPLC column conditions are as follows:

[0288] Purification at pH=2: Waters Sunfire TM C 18A 5 μm, 19 x 100 mm column was used for elution with mobile phase A: 0.1% TFA (trifluoroacetic acid) aqueous solution and mobile phase B: acetonitrile; the flow rate was 30 mL / min, and the separation gradient was optimized for each compound using a compound-specific method optimization scheme as described in the literature [see "Preparative LCMS Purification: Improved Compound Specific Method Optimization", K. Blom, B. Glass, R. Sparks, A. Combs, J. Comb. Chem., 6, 874-883 (2004)]. Typically, a flow rate of 60 mL / min was used with a 30 x 100 mm column.

[0289] Purification at pH=10: Waters XBridge C 18 A 5 μm, 19 x 100 mm column was used for elution with mobile phase A: 0.15% NH4OH aqueous solution and mobile phase B: acetonitrile; the flow rate was 30 mL / min, and the separation gradient was optimized for each compound using a compound-specific method optimization scheme as described in the literature [see "Preparative LCMS Purification: Improved Compound Specific Method Optimization", K. Blom, B. Glass, R. Sparks, A. Combs, J. Comb. Chem., 6, 874-883 (2004)]. Typically, a flow rate of 60 mL / min was used with a 30 x 100 mm column.

[0290] Example 1

[0291] Synthesis of 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-8-(morpholin-4-ylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (compound 1)

[0292]

[0293] Step 1: Synthesis of 4-((4-chloro-5-(1,3-dioxacyclopentan-2-yl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)morpholine

[0294] Under N2 atmosphere, 4-chloro-5-(1,3-dioxacyclopentan-2-yl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (50.0 g, 137 mmol) (see, for example, Example 2) and tetrahydrofuran (THF, 266 g, 300 mL) were added to a 1-L flask. A 2.0 M solution of lithium diisopropylamide in THF / heptane / phenylethane (77.4 g, 95 mL, 190 mmol, 1.4 equivalents) was added to this mixture at -70 °C. The mixture was stirred at -70 °C for 1 h. A THF solution of N-formylmorpholine (29.7 g, 258 mmol, 1.9 equivalents) (22.2 g, 25 mL) was added dropwise to the mixture. The reaction was carried out over 30 min after the addition. LC / MS showed clean formation of the desired product, 4-chloro-5-(1,3-dioxacyclopentan-2-yl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine-2-carboxaldehyde. The reaction was stopped with acetic acid (16.4 g, 15.6 mL, 274 mmol, 2.0 equivalent) and cooled with dry ice. Morpholine (33.7 g, 33.5 mL, 387 mmol, 2.83 equivalent) and acetic acid (74.0 g, 70 mL, 1231 mmol, and 9.0 equivalent) were added sequentially to the mixture at 0 °C (internal temperature increased from 0 °C to 18 °C) and the mixture was stirred overnight. Sodium triacetoxyborohydride (52.50 g, 247.7 mmol, 1.8 equivalent) was added and the temperature of the reaction mixture was increased from 20 °C to 32 °C. The mixture was stirred at room temperature for 30 min. HPLC and LC / MS indicated that the reaction was complete. Water (100 g, 100 mL) and an aqueous solution of 2.0 M sodium carbonate (Na₂CO₃) (236 g, 200 mL, 400 mmol, 2.9 equivalence) were added slowly in sequence (gas was expelled!). The mixture was stirred for about 30 min. The organic layer was separated and water (250 g, 250 mL) and heptane (308 g, 450 mL) were added. The resulting slurry was stirred for 1 h and the solid was collected by filtration. The wet filter cake was washed twice with heptane (75.00 mL x 2, 51.3 g x 2) and then dried overnight in an oven at 50 °C to give the desired product 4-((4-chloro-5-(1,3-dioxanepent-2-yl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)morpholine (52.00 g, 81.8% yield) as a light brown solid: C 21 H 23 LCMS calculated value of ClN2O5S [M+H] + : 464.00; Experimental value: 464.0; 1H NMR(400MHz,DMSO-d6)δ8.48(s,1H),8.38(m,2H),7.72(m,1H),7.64(m,2H),6.83(s ,1H),6.13(s,1H),4.12(m,2H),4.00(m,2H),3.92(s,2H),3.55(m,4H),2.47(m,4H).

[0295] Step 2: Synthesis of 4-chloro-2-(morpholinylmethyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde

[0296] 4-((4-chloro-5-(1,3-dioxacyclopentan-2-yl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)morpholine (20.00 g, 43.1 mmol) and dichloromethane (265 g, 200 mL) were charged into a 2 L reactor equipped with a thermocouple, a feeding funnel, and a mechanical stirrer at room temperature. The resulting mixture was stirred at room temperature (internal temperature 19.5 °C) to obtain a solution. Hydrochloric acid aqueous solution (0.5 M, 240 g, 200.0 mL, 100 mmol, 2.32 equivalents) was added to the resulting solution over 7 min at room temperature. After stirring at room temperature for more than 23 h, the bilayer reaction mixture transformed into a concentrated colorless suspension. When HPLC indicated the reaction was complete, the slurry was cooled to 0–5 °C and an aqueous sodium hydroxide solution (1 N, 104 g, 100 mL, 100 mmol, and 2.32 equivalents) was added over approximately 10 min to adjust the pH of the reaction mixture to 10–11. Heptane (164 g, 240 mL) was added and the reaction mixture was stirred at room temperature for 1 h. The solid was collected by filtration, and the wet filter cake was washed with water (2 x 40 mL) and heptane (2 x 40 mL), then dried under vacuum at 50 °C in an oven to provide the desired product, 4-chloro-2-(morpholinylmethyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde (16.9 g, 93% yield), as a light brown solid. 19 H 19 LCMS calculated value of ClN3O4S [M+H] + : 420.00; Experimental value: 420.0; 1 H NMR (400MHz, DMSO-d6) δ10.33(s,1H),8.76(s,1H),8.42(m,2H),7.74(m,1H),7.65(m,2H),6.98(s,1H),3.96(m,2H),3.564(m,4H),2.51(m,4H).

[0297] Step 3: Synthesis of N-((4-chloro-2-(morpholinylmethyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-2,6-difluoro-3,5-dimethoxyaniline

[0298] N,N-dimethylformamide (450 mL, 425 g), 4-chloro-2-(morpholinylmethyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde (30.0 g, 71.45 mmol), and 2,6-difluoro-3,5-dimethoxyaniline (14.2 g, 75.0 mmol) were added dropwise to this suspension (internal temperature 20-23 °C) at room temperature (internal temperature 20 °C) over 10 min. The suspension became a solution within 5 min after the addition of chlorotrimethylsilane. The solution was stirred at room temperature for 1.5 h and then cooled to 0-5 °C in an ice bath. A THF solution of the borane-THF complex (1.0 M, 71.4 mL, 71.4 mmol, 64.2 g, 1.0 equivalent) was added dropwise over 30 min via another funnel while maintaining the temperature at 0–5 °C. After the addition, the mixture was stirred for 4 h. Water (150 g, 150 mL) was added over 20 min under ice bath cooling, followed by slow addition of ammonium hydroxide solution (28% NH3, 15.3 g, 17 mL, 252 mmol, 3.53 equivalent) to pH 9–10 while maintaining the temperature below 10 °C. Water (250 mL, 250 g) was added again via another funnel. The slurry was stirred for 30 min and the solids were collected by filtration. The wet filter cake was washed with water (90 g x 2, 90 mL x 2) and heptane (61.6 g x 2, 90 mL x 2). The product was dried overnight by suction to obtain the desired product N-((4-chloro-2-(morpholinylmethyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-2,6-difluoro-3,5-dimethoxyaniline (41.6 g, 96% yield): C 27 H 28 LCMS calculated value of ClF₂N₄O₅S [M+H] + : 593.10; Experimental value: 593.1; 1H NMR(400MHz,DMSO-d6)δ8.36(m,2H),8.28(s,1H),7.72(m,1H),7.63(m,2H),6.78(s,1H),6 .29(m,1H),5.82(m,1H),4.58(m,2H),3.91(s,2H),3.76(s,6H),3.56(m,4H),2.47(m,4H).

[0299] Step 4: Synthesis of 1-((4-chloro-2-(morpholinylmethyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-1-(2,6-difluoro-3,5-dimethoxyphenyl)-3-ethylurea

[0300] N-((4-chloro-2-(morpholinylmethyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-2,6-difluoro-3,5-dimethoxyaniline (67.0 g, 113 mmol) and acetonitrile (670 mL, 527 g) were added to a 2-L 3-necked round-bottom flask equipped with a thermocouple, a nitrogen bubbler inlet, and a magnetic stirrer. The suspension was cooled to 0–5 °C. Ethyl isocyanate (17.7 mL, 15.9 g, 224 mmol, 1.98 equivalents) was added to the mixture over 30 seconds. The temperature was kept constant at 0.7 °C after the addition. Methanesulfonic acid (16.1 mL, 23.9 g, 248 mmol, 2.2 equivalents) was added dropwise over 35 min while maintaining the temperature below 2 °C. The mixture was heated to room temperature and stirred overnight. At 24 h after addition, the product content was 93.7%, unreacted SM was 0.73%, and the major impurity (diisocyanate adduct) was 1.3%. The mixture was cooled in an ice bath and quenched sequentially with sodium hydroxide (NaOH) solution (1.0 M, 235 mL, 244 g, 235 mmol, 2.08 equivalents) for 20 min and with saturated sodium bicarbonate (NaHCO3) aqueous solution (1.07 M, 85 mL, 91 g, 0.091 mol, 0.80 equivalents) for 10 min. Water (550 mL, 550 g) was added, and the liquid became a single phase. The mixture was stirred for 2 h and the solid was collected by filtration and washed with water (165 mL, 165 g) to give 1-((4-chloro-2-(morpholinylmethyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-1-(2,6-difluoro-3,5-dimethoxyphenyl)-3-ethylurea (70.3 g, 93.7% yield).

[0301] Crude 1-((4-chloro-2-(morpholinylmethyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-1-(2,6-difluoro-3,5-dimethoxyphenyl)-3-ethylurea (68.5 g, 103 mmol) was added to acetonitrile (616 mL, 485 g). The mixture was heated to 60–65 °C to obtain a dilute amber suspension. The solid was filtered off with diatomaceous earth, which was then washed with acetonitrile (68.5 mL, 53.8 g). Water (685 g, 685 mL) was added to the pale yellow filtrate to form a slurry. The slurry was stirred overnight at room temperature and filtered. The solid was added to water (685 mL, 685 g) and stirred at 60 °C for 2 h. The solid was filtered off and slurryed again in heptane (685 mL, 469 g) overnight. The product was dried in an oven at 50°C under vacuum for 48 h to provide 1-((4-chloro-2-(morpholinylmethyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-1-(2,6-difluoro-3,5-dimethoxyphenyl)-3-ethylurea (62.2 g, 90.8% yield, 99.9% purity according to HPLC area %). KF was 0.028%. Acetonitrile (according to 1 The H NMR (H NMR) was approximately 1.56%, and the DCM (according to 1 2.0% (HNMR): C 30 H 33 LCMS calculated value of ClF2N5O6S [M+H] + EM: 664.17; Experimental value: 664.2; 1 H NMR(400MHz,DMSO-d6)δ8.33(m,2H),8.31(s,1H),7.72(m,1H),7.64(m,1H),6.96(m,2H),6.73(s,1H),6 .43(m,1H),4.87(s,2H),3.90(s,2H),3.77(s,6H),3.54(m,4H),3.03(m,2H),2.46(m,4H),0.95(m,3H).

[0302] Step 5: Synthesis of 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-8-(morpholin-4-ylmethyl)-7-(phenylsulfonyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one

[0303] A 2000 mL flask equipped with a thermocouple, nitrogen inlet, and mechanical stirrer was loaded with dry 1-((4-chloro-2-(morpholinylmethyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-1-(2,6-difluoro-3,5-dimethoxyphenyl)-3-ethylurea (30.0 g, 45.2 mmol, KF = 0.11%) and tetrahydrofuran (1200 mL, 1063 g). A 1.0 M THF solution of hexamethyldisilazide (62.3 mL, 55.5 g, 62.3 mmol, 1.38 equivalents) was added to this suspension at room temperature. After the addition of the base, the mixture became a solution. The reaction mixture was stirred for 2 h, and HPLC showed that the starting material was undetectable. Add 1.0 M hydrochloric acid (18.1 mL, approx. 18.1 g, 18.1 mmol, 0.4 equivalents) to this mixture. Concentrate the solution to 600 mL and add water (1200 mL, 1200 g). A slurry is formed after the addition of water. Stir the slurry at room temperature for 30 min and collect the solid by filtration. Wash the wet filter cake twice with water (60 mL x 2, 60 g x 2) and dry overnight at 50 °C to give a light brown solid of 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-8-(morpholin-4-ylmethyl)-7-(phenylsulfonyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (26.58 g, 93.7% yield at original rate): THF according to 1 The H NMR was 0.32%, KF was 5.26%, and the adjusted yield was 88.5%. 30 H 32 LCMS calculated value of F2N5O6S [M+H] + EM: 628.20; Experimental value: 628.2; 1 H NMR(400MHz,DMSO-d6)δ8.41(m,2H),8.07(s,1H),7.70(m,1H),7.63(m,2H),7.05(m,1H),6.89(s ,1H),4.76(s,2H),4.09(m,2H),3.93(s,2H),3.89(s,6H),3.60(m,4H),2.50(m,4H),1.28(m,3H).

[0304] Step 6: Synthesis of 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-8-(morpholin-4-ylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one

[0305] A 1M aqueous solution of sodium hydroxide (63.7 mL, 66.3 g, 63.7 mmol) was added to a stirred suspension of 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-8-(morpholinylmethyl)-7-(phenylsulfonyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (10.0 g, 15.93 mmol) in 1,4-dioxane (100 mL, 103 g). The reaction mixture was heated at 75 °C for 18 h. LC-MS showed that the reaction was complete. Water (100 mL, 100 g) was added to obtain a concentrated suspension. This slurry was stirred at room temperature for 1 h and filtered. The filter cake was washed with water (3 x 10 mL, 3 x 10 g) and heptane (2 x 10 mL, 2 x 6.84 g). The filter cake was dried overnight under vacuum by vacuum through the filter cake and then dried overnight under vacuum at 50 °C in an oven to give 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-8-(morpholin-4-ylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (6.8 g, 87.6% yield): C 24 H 28 LCMS calculated value of F2N5O4 [M+H] + : 488.20; Experimental value: 488.2.

[0306] Example 2. Synthesis of 4-chloro-5-(1,3-dioxacyclopentan-2-yl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine

[0307]

[0308] Step 1: Synthesis of 1H-pyrrolo[2,3-b]pyridine-7-oxide

[0309] m-Cloroperoxybenzoic acid (105.5 kg, 612 mol, 1.2 equivalents) was added to a solution of 1H-pyrrolo[2,3-b]pyridine (60 kg, 507.6 mol) in dichloromethane (600 L) for 5 h at 0–10 °C with stirring. After the addition was complete, the mixture was stirred at 0–10 °C for 3 h. The resulting solid was collected by filtration, washed with heptane, and dried to give 1H-pyrrolo[2,3-b]pyridine-7-oxide. The mother liquor was concentrated and the residue was treated with dichloromethane:heptane (2:3) and filtered to recover additional substances. Crude 1H-pyrrolo[2,3-b]pyridine-7-oxide (72 kg, 96% purity) was obtained and used unpurified for the next step.

[0310] Step 2: Synthesis of 4-chloro-1H-pyrrolo[2,3-b]pyridine

[0311] Crude 1H-pyrrolo[2,3-b]pyridine-7-oxide (72 kg, 253 mol) was dissolved in DMF (360 L) and heated at 50 °C. A solution of methanesulfonyl chloride (85.2 kg, 746 mol, 3.0 equivalent) was added dropwise to the solution while maintaining the temperature below 70 °C. After stirring at 90 °C for 2 h, the reaction solution was cooled to room temperature and added to 720 kg of ice / water. The mixture was neutralized with 6.0 M NaOH at 0 °C. The resulting precipitate was collected by filtration and washed with water. The solid was mixed with 72 L of water, 48 L of ethanol, and 29 L of 30% NaOH and stirred at room temperature for 1–2 h. Water (144 L) was added, and the mixture was treated with 37% HCl to adjust the pH to approximately 1. The product was collected by filtration and dried to obtain 4-chloro-1H-pyrrolo[2,3-b]pyridine (crude 26 kg, 97% purity, used unpurified): 1 HNMR (400MHz, CDCl3) δ11.30(s,1H),8.25(m,1H),7.44(m,1H),7.16(m,1H),6.65(m,1H).

[0312] Step 3: Synthesis of 4-chloro-1-(triisopropylsilyl)-1H-pyrrolo[2,3-b]pyridine

[0313] A solution of crude 4-chloro-1H-pyrrolo[2,3-b]pyridine (24 kg, 155.2 mol) in 216 L of THF was stirred at 0 °C while NaH (60%, 7.56 kg, 188.6 mol, 1.3 equivalents) was added dropwise under N2. After the addition, the mixture was stirred at room temperature for 1 h. Triisopropylsilyl chloride (39.6 kg, 188.6 mol, 1.3 equivalents) was added dropwise while maintaining the temperature below 25 °C. After stirring for 20 h, the mixture was quenched with 144 L of water and extracted with 144 L of heptane. The aqueous layer was back-extracted with 72 L of methyl tert-butyl ether. The combined organic layers were dried over anhydrous MgSO4 and concentrated under vacuum to obtain crude 4-chloro-1-(triisopropylsilyl)-1H-pyrrolo[2,3-b]pyridine in liquid form. The substance is used without purification, but its water content is controlled to be below 0.1%.

[0314] Step 4: Synthesis of 4-chloro-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde

[0315] Crude 4-chloro-1-(triisopropylsilyl)-1H-pyrrolo[2,3-b]pyridine (50 kg, approximately 138 mol) and anhydrous THF (150 kg) were charged into a 1000 L cryogenic reactor. The mixture was cooled to -75 °C and stirred under N2 while S-BuLi (1.3 M cyclohexane solution, 230 L, 300 mol, 2.2 equivalents) was added dropwise over approximately 6.0 h, maintaining the internal temperature below -60 °C. The mixture was stirred again at -75 °C for 2 h. N,N-dimethylformamide (30.4 kg, 416.1 mol, 3.0 equivalents) was added dropwise over approximately 3.0 h to maintain the internal temperature below -65 °C. After stirring at -65 to -75°C for 2 hours, the mixture was quenched by dropwise addition of a solution of 20% HCl in isopropanol (115 kg, 635 mol, 4.5 equivalents). The mixture was then stirred overnight at room temperature (20–25°C). The pH was adjusted to 7–8 by adding saturated NaHCO3. The precipitate was collected by filtration. The filter cake was washed with 76 L of water to give 4-chloro-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde (14 kg, 58% yield): 1 HNMR (400MHz, DMSO-d6) δ12.54(s,1H),10.35(s,1H),8.67(s,1H),7.74(m,1H),6.72(m,1H).

[0316] Step 5: Synthesis of 4-chloro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde

[0317] N,N-dimethylformamide (108 L) and 4-chloro-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde (10.8 kg, 59.8 mol) were charged into a 500 L reactor and cooled to 0–5 °C. Cesium carbonate (39 kg, 120 mol) was added to the resulting concentrated slurry at 0–5 °C. The slurry was stirred at 0 °C for approximately 20 min until the mixture became an amber-colored thin slurry. Benzenesulfonyl chloride (11.6 kg, 65.8 mol, 1.1 equivalents) was added dropwise to the thin slurry through a feeding funnel below 10 °C. The resulting slurry was stirred below 10 °C for 1 h, and HPLC indicated the reaction was complete. Extended stirring overnight at room temperature had almost no effect on the morphology of the reaction mixture. Water (160 L) was added to this mixture, and the slurry was stirred for 1 h. The solids were collected by filtration (slowly). The filter cake was washed with water and dried under vacuum in an oven to give 17.8 kg of 4-chloro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde (93% yield) as a light brown solid: C 14 H 10LCMS calculated value of ClN2O3S [M+H] + : 321.00; Experimental value: 320.9; 1 HNMR(400MHz, DMSO-d6)δ:10.34(s,1H),8.78(s,1H),8.18(m,3H),7.77(m,1H),7.66(m,2H),7.05(m,1H).

[0318] Step 6: Synthesis of 4-chloro-5-(1,3-dioxacyclopentan-2-yl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine

[0319] Toluene (270 L), 4-chloro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde (27 kg, 84.2 mmol), p-toluenesulfonic acid monohydrate (217 g, 1.26 mol, 0.015 equivalents), and 1,2-ethylene glycol (73.7 kg, 1187 mol, 14.1 equivalents) were charged into a 1000 L reactor. The mixture was stirred and heated to reflux to remove water (some ethylene glycol was also removed as the reaction progressed) for 9 h (LCMS showed the reaction was complete). After stirring overnight at room temperature, the mixture was diluted with ethyl acetate (135 L) and washed with a saturated NaHCO3 solution. The layers were separated and the organic layer was washed with a 10% NaCl aqueous solution and concentrated. Heptane (108 L) was added to form a slurry. The solid was collected by filtration. The solid was dissolved in dichloromethane (108 L) and filtered to remove mechanical impurities. The filtrate was concentrated and then dissolved in 67.5 L (2.5 V) of hot ethyl acetate and stirred for 2 h. The mixture was cooled while the solid formed. The solid was collected by filtration to give 4-chloro-5-(1,3-dioxacyclopentan-2-yl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine as a grayish-white solid (22 kg, 70% yield): C 16 H 14 LCMS calculated value of ClN₂O₄S [M+H] + : 365.03; Experimental value: 365.1; 1 H NMR (400MHz, DMSO-d6) δ8.51(s,1H),8.13(m,2H),8.07(m,1H),7.73(m,1H),7.63(m,2H),6.90(m,1H),6.13(s,1H),4.12(m,2H),3.98(m,2H).

[0320] Example 3. Substitute Synthesis of 4-chloro-2-(morpholin-4-ylmethyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde

[0321]

[0322] Step 1: Synthesis of 5-bromo-4-chloropyridine-2-amine

[0323] N-bromosuccinimide (131.5 g, 738.9 mmol, 0.95 equivalent) was added partically to a slurry of 2-amino-4-chloropyridine (100 g, 777.8 mmol, 1.0 equivalent) in acetonitrile (500 mL, 5 relative volumes) at 15–20 °C for 2 h. The reaction mixture was stirred for 30 min and the conversion was verified by HPLC. Based on the conversion, 0–5 mol% of N-bromosuccinimide was added and the mixture was stirred for another 15 min. After HPLC indicated that the conversion was complete, the reaction mixture was heated and acetonitrile (300 mL) was distilled off at atmospheric pressure. Water (250 mL) was added and the temperature was adjusted to 50–55 °C to form a slurry. The resulting slurry was stirred for 30 min and water (350 mL) was added over 1 h. The slurry was cooled to 20–25 °C, stirred for 1 h, and the solid was collected by filtration. The wet filter cake was washed with a mixture of water (75 mL) and acetonitrile (25 mL) to give the wet product 5-bromo-4-chloropyridin-2-amine (191 g, purity 92.1% based on HPLC area %). The wet product was dissolved in acetic acid (500 mL, based on 5 relative volumes of 2-amino-4-chloropyridinium, 55–70 °C) and the solution was used directly for the next step.

[0324] Step 2: Synthesis of 5-bromo-4-chloro-3-iodopyridine-2-amine

[0325] A solution of 5-bromo-4-chloropyridin-2-amine in acetic acid (500 mL acetic acid containing 191 g of 5-bromo-4-chloropyridin-2-amine) was distilled under reduced pressure at 40–60 °C to remove the solvent. Then, sulfuric acid (39.7 g, 96% w / w, 388.9 mmol, 0.5 equivalents) and iodine (76.2 g, 300.3 mmol, 0.386 equivalents) were added, and the temperature was adjusted to 77–83 °C. At this temperature, a solution of periodic acid (50% w / w, 54.89 g, 120.4 mmol, 0.155 equivalents) was added over 2–3 h. The reaction mixture was stirred at 77–83 °C for 2–3 h, and the conversion was verified by HPLC (SM < 1.0% a / a). At 75-85°C, quench the reaction mixture by adding solid ammonium sulfite in parts of 4.53 g (0.05 equivalents) until the KI / starch test is negative. Typically, two parts (0.1 equivalents) of ammonium sulfite are required. The end of quenching can also be observed by the presence of a purple color in the absence of iodine. Then, dilute the reaction mixture with water (200 mL, 2.0 relative volume, at room temperature) and lower the temperature to about 50°C. Allow the product to precipitate. Adjust the pH to 3.0-3.5 at 45-60°C with ammonia (required to be 25%-w / w in water, about 63.6 g, 0.93 mol, 1.2 equivalents). Neutralization is strongly exothermic. Stir the slurry at 45-50°C for 30 min and then collect the solids by filtration. Wash the filter cake with typically about 600 mL of water and then with 2-propanol (200 mL). The wet product was dried in a vacuum chamber at 60 °C to give 213.5 g (82.3% yield) of 5-bromo-4-chloro-3-iodopyridin-2-amine as a yellow to beige solid: LCMS calculated value of C5H4BrClIN2 [M+H] + : 332.82; Experimental value: 332.8; 1 H NMR (400MHz, DMSO-d6) δ8.09 (s, 1H), 6.60 (s, 2H).

[0326] Step 3: Synthesis of 5-bromo-4-chloro-3-(3-morpholinylprop-1-yn-1-yl)pyridine-2-amine

[0327] 5-Bromo-4-chloro-3-iodopyridin-2-amine (50 g, 150 mmol, 1.0 equivalent), 4-(prop-2-ynyl)morpholine (22.5 g, 180 mmol, 1.20 equivalent), diisopropylamine (18.2 g, 180 mmol, 1.2 equivalent), and 150 mL of toluene were charged into the reactor. The solution was carefully degassed using three vacuum argon cycles. Then, CuI (0.29 g, 1.5 mmol, 1 mol%) and Pd(PPh3)4 were added, and the flask was purged again with argon. The mixture was stirred overnight (17 h) at 50 °C. Water (50 mL, 1 volume) was added in bulk, and the mixture was cooled to 20–25 °C. The crude product was filtered off and washed continuously with 10% ammonia (50 ml, 1.0 v / v), water (50 ml, 1 v / v), toluene (25 ml, 0.5 v / v), and 2-isopropanol (50 ml, 1.0 v / v). After drying under vacuum at 50 °C, 5-bromo-4-chloro-3-(3-morpholinylprop-1-yn-1-yl)pyridine-2-amine (41.6 g, 87% yield) was obtained as a light brown solid. 12 H 14 LCMS calculated value of BrClIN5O [M+H] + : 329.99; Experimental value: 330.0; 1 H NMR (400MHz, DMSO-d6) δ8.13(s,1H),6.69(s,2H),3.64(s,2H),3.61(m,4H),2.54(m,4H).

[0328] Step 4: Synthesis of 4-((5-bromo-4-chloro-1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)morpholine

[0329] A solution of KOtBu (18.1 g, 1.4 equivalents, 112.21 mmol) in tetrahydrofuran (114 mL, 3 volumes) was heated to 30–35 °C, while 5-bromo-4-chloro-3-(3-morpholinylprop-1-yn-1-yl)pyridin-2-amine (38 g, 114.9 mmol, 1.0 equivalents) was added partically at 30–35 °C for 1.0 h. After stirring for 2 h, the reaction was stopped with a solution of acetic acid (10.4 g, 172.4 mmol, 1.5 equivalents) in water (76 mL, 2 volumes), and 76 mL of THF was removed by distillation. The solution was then heated to reflux and MeOH (38 mL, 1 volume) was added, and the resulting suspension was cooled to 23 °C for 1 h. After stirring at 23°C for 0.5 h, the solid was filtered off and washed with water (38 mL, 1 volume) and MeOH (38 mL, 1 volume). After drying under vacuum at 50°C, 4-((5-bromo-4-chloro-1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)morpholine (32.8 g, 86% yield) was obtained as a light brown powder. 12 H 14 LCMS calculated value of BrClIN5O [M+H] + : 329.99; Experimental value: 329.8; 1 H NMR (400MHz, DMSO-d6) δ12.22(s,1H),8.34(s,1H),6.40(s,1H),3.65(s,2H),3.58(m,4H),2.42(m,4H).

[0330] Step 5: Synthesis of 4-((5-bromo-4-chloro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)morpholine

[0331] A slurry of 4-((5-bromo-4-chloro-1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)morpholine (10 g, 30.25 mmol, 1.0 equivalent, analytical 94% w / w) and NaH (1.69 g, 60%, 42.35 mmol, 1.4 equivalent) in 38 mL of tetrahydrofuran was cooled to 0–5 °C, while PhSO₂Cl (7.48 g, 42.35 mmol, 1.4 equivalent) was added over 1 h. After 1.5 h, HPLC indicated incomplete reaction. NaH (0.34 g, 0.3 equivalent) was added again, at which point gas escape was observed. When HPLC indicated completion, the reaction mixture was quenched with a mixture of acetic acid (0.5 g) and water (15 mL) with methanol (15 mL). The pH was adjusted to 6.5 with sodium hydroxide, and the product was separated by filtration. The wet filter cake was washed with 2-isopropanol (20 mL) and water (20 mL), and the wet product (14.8 g) was dried in a vacuum chamber to give 4-((5-bromo-4-chloro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)morpholine (12.57 g, 86% yield) as a brown solid: C 18 H 18 LCMS calculated value of BrClIN3O3S [M+H] + : 469.99; Experimental value: 470.0; 1 H NMR (400MHz, DMSO-d6) δ8.56(s,1H),8.33(m,2H),7.73(m,1H),7.65(m,2H),6.83(s,1H),3.91(s,2H),3.53(m,4H),2.46(m,4H).

[0332] Step 6: Synthesis of 4-chloro-2-(morpholinylmethyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde

[0333] iPrMgCl (6.9 mL, 2 M tetrahydrofuran solution, 13.80 mmol, 1.3 equivalent) was added to a suspension of 4-((5-bromo-4-chloro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)morpholine (5.0 g, 10.6 mmol, 1.0 equivalent) in 50 mL of tetrahydrofuran at -10 °C to 0 °C. After stirring for 2 h, N,N-dimethylformamide (1.55 g, 21.2 mmol, 2.0 equivalent) was added to the reaction solution at -5 °C to 0 °C for 0.5 h. The mixture was stirred at -5 °C to 0 °C for 0.5 h, then heated to 23 °C for 0.5 h and stirred at 23 °C for 1 h. The pH was adjusted to 6-7 by adding 1.5 mL of acetic acid and 10 mL of water. 25 mL of MeOH and 15 mL of water were added to the biphase mixture. After stirring for 1 h, the product was filtered and washed with 20 mL MeOH / water (1 / 1) and 30 mL water. After drying under vacuum at 50 °C, 3.39 g (76% yield) of 4-chloro-2-(morpholinylmethyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde as a grayish-white powder was obtained: C 19 H 19 LCMS calculated value of ClN3O4S [M+H] + : 420.07; Experimental value: 420.0; 1 H NMR (400MHz, DMSO-d6) δ10.33(s,1H),8.76(s,1H),8.42(m,2H),7.74(m,1H),7.65(m,2H),6.98(s,1H),3.96(m,4H),3.564(m,4H),2.51(m,4H).

[0334] Example 4.2 Synthesis of 6-difluoro-3,5-dimethoxyaniline

[0335]

[0336] Step 1: Synthesis of methyl pentafluorobenzoate

[0337] SOCl2 (29.2 kg, 245.2 mol, 1.3 equivalents) was added dropwise to a solution of pentafluorobenzoic acid (40 kg, 188.6 mol) in 68 L of methanol at 20–50 °C for 4.0 h. The mixture was then heated to reflux for 17 h. Methanol was removed by vacuum distillation, and the residue was dissolved in methyl tert-butyl ether (77 L). The solution was washed with saturated NaHCO3 (37 L), dried over MgSO4, and evaporated to give methyl pentafluorobenzoate (39 kg, 91% yield) as a colorless oil. 1H NMR (400MHz, CDCl3) δ3.90 (s, 3H).

[0338] Step 2: Synthesis of methyl 4-(phenylmethylamino)-2,3,5,6-tetrafluorobenzoate

[0339] Methyl pentafluorobenzoate (39 kg, 172.5 mol) and N,N-diisopropylethylamine (26.8 kg, 207 mol, 1.2 equivalents) were dissolved in N-methylpyrrolidone (39 L). Acetophenone (18.5 kg, 172.5 mol, 1.0 equivalent) was added dropwise to 19.5 L of N-methylpyrrolidone solution over 3.5 h, while maintaining the internal temperature below 50 °C. The resulting concentrated yellow slurry was heated to 65 °C and stirred for another 1 h. The mixture was poured into 195 L of aqueous acetic acid solution (10% acetic acid and 90% H₂O), and the slurry was stirred for 1 h and filtered. The filter cake was washed with water and heptane and dried under vacuum at 35 °C to give methyl 4-(benzylamino)-2,3,5,6-tetrafluorobenzoate (38 kg, 70% yield). 1 H NMR (400MHz, CDCl3) δ7.37(m,5H),4.67(m,2H),4.58(m,1H),3.93(s,3H).

[0340] Step 3: Synthesis of 4-(benzylamino)-3,5-difluoro-2,6-dimethoxybenzoic acid

[0341] Methanol (72 L) containing methyl 4-(phenylmethylamino)-2,3,5,6-tetrafluorobenzoate (38 kg, 121.3 mol) was stirred under N2 at room temperature while a solution of NaOMe in methanol (25 wt%, 110.8 kg, 545.85 mol, 4.5 equivalents) was added dropwise over 3.0 h, maintaining the temperature below 50 °C. After heating to 65–70 °C for 18 h, 18 L of water was added to the reaction mixture and the resulting solution was stirred for 1 h. The solvent was removed by vacuum distillation. Water (54 L) was added and the resulting solution was acidified to pH 2 with 37% HCl. The mixture was extracted three times with ethyl acetate (54 kg each time). The combined organic extracts were washed with water (43 L) and concentrated to dryness to form a solid. The solid was wet-milled with heptane (43 L) to remove impurities. The solid was collected and dried under vacuum at 40 °C to give 4-(benzylamino)-3,5-difluoro-2,6-dimethoxybenzoic acid (35 kg, 86% yield): 1 H NMR (400MHz, CDCl3) δ12.74(s,1H),7.37(m,5H),6.62(s,1H),4.67(m,2H),3.96(s,6H).

[0342] Step 4: Synthesis of n-benzyl-2,6-difluoro-3,5-dimethoxyaniline

[0343] 4-(phenylmethylamino)-3,5-difluoro-2,6-dimethoxybenzoic acid (17 kg) was heated to 75-85 °C for 3-4 h under pristine conditions. After the reaction was complete, 40 L of methyl tert-butyl ether and 20 L of 1 M NaOH were added. The mixture was stirred at room temperature for 30 min. The organic layer was separated and washed with water (20 L) and brine (20 L). The organic phase was concentrated under reduced pressure to give a crude product. The crude product was wet-milled with heptane and dried under vacuum at 35 °C to give N-phenylmethyl-2,6-difluoro-3,5-dimethoxyaniline (12 kg, 82% yield). 1 H NMR (400MHz, CDCl3) δ7.35(m,5H),6.09(m,1H),4.53(m,2H),4.00(s,1H),3.85(s,6H).

[0344] Step 5: Synthesis of 2,6-difluoro-3,5-dimethoxyaniline

[0345] N-phenylmethyl-2,6-difluoro-3,5-dimethoxyaniline (24 kg, 85.9 mol) was dissolved in a mixed solvent of ethanol (120 L) and acetic acid (20 L), with ammonium formate (13.2 kg) and 1.68 kg of Pd / C added simultaneously. The mixture was heated at 50 °C for 2–3 h. The reaction mixture was then subjected to… The mixture was filtered, washed with ethanol (1.2 L x 2), and concentrated. The crude material was added to 80 L of water, and the resulting slurry was filtered. The crude material was added to 60 L of methyl tert-butyl ether and 2.5 kg of activated carbon, and the mixture was heated to reflux for 3 h. After filtration and concentration, the resulting solid was added to 36 L of heptane and stirred at room temperature for 2 h. The mixture was filtered and dried under vacuum at 35 °C to give 2,6-difluoro-3,5-dimethoxyaniline (15.2 kg, 93% yield) as a light brown solid: C8H 10 LCMS calculated value of F2NO2 [M+H] + : 190.16; Experimental value: 190.1; 1 H NMR (400MHz, DMSO-d6) δ6.16(m,1H),5.18(s,2H),3.78(s,6H).

[0346] Example 5.3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-8-(morpholin-4-ylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (compound 1) substitutional synthesis

[0347]

[0348] Step 1: 4-(ethylamino)-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde

[0349]

[0350] A mixture of 4-chloro-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde (catalog number 958230-19-8, Lakestar Tech, batch: 124-132-29: 3.0 g, 17 mmol) and ethylamine (10 M aqueous solution, 8.3 mL, 83 mmol) in 2-methoxyethanol (20 mL, 200 mmol) was heated to 130 °C and stirred overnight. The mixture was cooled to room temperature and then concentrated under reduced pressure. The residue was treated with 1 N HCl (30 mL) and stirred at room temperature for 1 h, then neutralized with a saturated aqueous solution of NaHCO3. The precipitate was collected by filtration, washed with water, and dried to provide the desired product (2.9 g, 92%). 10 H 12 LC-MS calculated value of N3O [M+H] + m / z: 190.1; Experimental value: 190.1.

[0351] Step 2: 5-{[(2,6-difluoro-3,5-dimethoxyphenyl)amino]methyl}-N-ethyl-1H-pyrrolo[2,3-b]pyridine-4-amine

[0352]

[0353] A mixture of 4-(ethylamino)-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde (7.0 g, 37 mmol), 2,6-difluoro-3,5-dimethoxyaniline (9.1 g, 48 mmol), and [(1S)-7,7-dimethyl-2-oxobicyclo[2.2.1]hept-1-yl]methanesulfonic acid (Aldrich, catalog number 21360: 2 g, 7 mmol) in xylene (250 mL) was heated to reflux, and water was removed by azeotropic extraction using a Dean-Stark apparatus for 2 days, at which point LC-MS showed the reaction was complete. The mixture was cooled to room temperature, and the solvent was removed under reduced pressure. The residue was dissolved in tetrahydrofuran (500 mL), and then 2.0 M lithium tetrahydroaluminate THF solution (37 mL, 74 mmol) was slowly added. The resulting mixture was stirred at 50 °C for 3 h, and then cooled to room temperature. The reaction was stopped by adding water, a 15% NaOH aqueous solution, and water. The mixture was filtered and washed with THF. The filtrate was concentrated and the residue was washed with CH2Cl2, then filtered to give the pure product (11 g, 82%). 18 H 21 LC-MS calculated values ​​of F₂N₄O₂ [M+H] + m / z: 363.2; experimental value: 363.1.

[0354] Step 3: 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one

[0355]

[0356] A solution of triphosgene (5.5 g, 18 mmol) in tetrahydrofuran (30 mL) was slowly added at 0 °C to a mixture of 5-{[(2,6-difluoro-3,5-dimethoxyphenyl)amino]methyl}-N-ethyl-1H-pyrrolo[2,3-b]pyridine-4-amine (5.6 g, 15 mmol) in tetrahydrofuran (100 mL), and the mixture was stirred at room temperature for 6 h. The mixture was cooled to 0 °C, and then a 1.0 M aqueous solution of sodium hydroxide (100 mL, 100 mmol) was slowly added. The reaction mixture was stirred overnight at room temperature, and the precipitate formed was collected by filtration, washed with water, and then dried to provide the first batch of purified desired product. The organic layer in the filtrate was separated, and the aqueous layer was extracted with methane chloride. The combined organic layers were concentrated, and the residue was wet-milled with methane chloride, then filtered and dried to provide another batch of product (total 5.5 g, 92%). 19 H 19 LC-MS calculated values ​​of F₂N₄O₃ [M+H]+ m / z: 389.1; Experimental value: 389.1.

[0357] Step 4: 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-7-(phenylsulfonyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one

[0358]

[0359] Sodium hydride (185 mg, 4.63 mmol in mineral oil) was added to a solution of 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (900 mg, 2.32 mmol) in N,N-dimethylformamide (20 mL). The resulting mixture was stirred at 0 °C for 30 min, and then benzenesulfonyl chloride (0.444 mL, 3.48 mmol) was added. The reaction mixture was stirred at 0 °C for 1.5 h, at which point LC-MS showed that the reactants were completely converted to the desired product. The reaction was stopped with saturated NH4Cl solution and diluted with water. The white precipitate was collected by filtration, washed with water and hexane, and dried to provide the desired product (1.2 g, 98%) as a white solid, which was used in the next step without further purification. 25 H 23 LC-MS calculated value of F2N4O5S [M+H] + m / z: 529.1; Experimental value: 529.1.

[0360] Step 5: 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-2-oxo-7-(phenylsulfonyl)-2,3,4,7-tetrahydro-1H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidine-8-carboxaldehyde

[0361]

[0362] Freshly prepared lithium diisopropylamide (1 M THF solution, 3.48 mL, 3.48 mmol) was added to a solution of 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-7-(phenylsulfonyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (1.75 g, 3.31 mmol) in tetrahydrofuran (80 mL). The resulting mixture was stirred at -78 °C for 30 min, and then N,N-dimethylformamide (1.4 mL, 18 mmol) was slowly added. The reaction mixture was stirred at -78 °C for 30 min, then the reaction was stopped with water and extracted with EtOAc. The organic extracts were combined and washed with water and brine. The organic layer was dried over Na2SO4 and concentrated. The residue was purified by rapid chromatography, eluting with 0 to 20% EtOAc / DCM to give the desired product (1.68 g, 91%) as a white solid. 26 H 23 LC-MS calculated values ​​(M+H) of F2N4O6S + m / z: 557.1; experimental value: 556.9.

[0363] Step 6: 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-8-(morpholin-4-ylmethyl)-7-(phenylsulfonyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one

[0364]

[0365] Morpholine (0.95 mL, 11 mmol) was added to a solution of 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-2-oxo-7-(phenylsulfonyl)-2,3,4,7-tetrahydro-1H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidine-8-carboxaldehyde (1.73 g, 3.11 mmol) in dichloromethane (50 mL), followed by the addition of acetic acid (2 mL, 30 mmol). The resulting yellow solution was stirred overnight at room temperature, followed by the addition of sodium triacetoxyborohydride (2.3 g, 11 mmol). The mixture was stirred at room temperature for 3 h, at which point LC-MS showed complete reaction of the reactants into the desired product. The reaction was stopped with saturated NaHCO3, followed by extraction with ethyl acetate (EtOAc). The organic extracts were combined and washed with water and brine. The organic layer was dried over Na2SO4 and concentrated. The residue was purified by rapid chromatography, eluting with 0 to 40% EtOAc / DCM to give the desired product (1.85 g, 95%) as a yellow solid. 30 H32 LC-MS calculated values ​​(M+H) of F2N5O6S + m / z: 628.2; Experimental value: 628.0.

[0366] Step 7: 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-8-(morpholin-4-ylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one

[0367] To a solution of 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-8-(morpholin-4-ylmethyl)-7-(phenylsulfonyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (1.5 g, 2.4 mmol) in tetrahydrofuran (40 mL), tetrabutylammonium fluoride (1 M THF solution, 7.2 mL, 7.2 mmol) was added. The resulting solution was stirred at 50 °C for 1.5 h, then cooled to room temperature and the reaction was stopped with water. The mixture was extracted with dichloromethane (DCM), and the organic extracts were combined and washed with water and brine. The organic layer was dried over Na₂SO₄ and concentrated. The residue was purified by rapid chromatography, eluting with 0 to 10% MeOH / DCM to obtain the desired product as a white solid. The desired product was further purified by preparative HPLC (pH = 2, acetonitrile / H₂O). 24 H 28 LC-MS calculated values ​​(M+H) of F2N5O4 + m / z: 488.2; Experimental value: 488.0. 1 H NMR (500MHz, DMSO) δ12.09(s,1H),8.06(s,1H),7.05(t,J=8.1Hz,1H),6.87(s,1H),4.78(s,2H),4.50(s,2H),4. 17(q,J=6.8Hz,2H),3.97(br,2H),3.89(s,6H),3.65(br,2H),3.37(br,2H),3.15(br,2H),1.37(t,J=6.8Hz,3H).

[0368] Example 6.7 Synthesis of (2,6-difluoro-3,5-dimethoxy-phenyl)-9-ethyl-2-morpholin-4-ylmethyl-8-oxo-6,7,8,9-tetrahydro-3,4,7,9-tetraaza-cyclopentadieno[a]naphthalene-3-sulfonic acid dimethylamide

[0369]

[0370] Step 1. Synthesis of 7-(2,6-difluoro-3,5-dimethoxy-phenyl)-9-ethyl-8-oxo-6,7,8,9-tetrahydro-3,4,7,9-tetraaza-cyclopentadieno[a]naphthalene-3-sulfonic acid dimethylamide

[0371]

[0372] A 200L glass reactor was assembled with an elevated stirred condenser, thermocouples, a feed funnel, and a nitrogen inlet, and the apparatus was purged with nitrogen. Drinking water (3.1L) and sodium hydroxide (3093g) were added to the reactor and the mixture was stirred at approximately 71°C until a solution was obtained. The reaction mixture was cooled to approximately 30°C and THF (15.0L) was added. The reaction mixture was cooled to 10°C and 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (3000g) and n-Bu4N were added sequentially. + HSO4 - (262g), while maintaining the temperature at approximately 10°C. The material was rinsed into the reactor with THF (15.0L), while maintaining the temperature at approximately 7°C. N,N-dimethylamine sulfonyl chloride (1.244L) was added, while maintaining the temperature at approximately 7°C. The reaction mixture was heated to approximately 17°C and stirred at approximately 22°C for 7 hours. Drinking water (120.0L) was added, while maintaining the temperature at approximately 20°C, and the reaction mixture was stirred at approximately 18°C ​​for 1 hour. The reaction mixture was filtered, and the filter cake was washed four times with drinking water (30.0L each time). The product was air-dried on a filter for 14.5 hours to provide crude 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-N,N-dimethyl-2-oxo-1,2,3,4-tetrahydro-7H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidine-7-sulfonamide (4694 g).

[0373] Purification: A 100L glass reactor was assembled with an elevated stirred condenser, thermocouples, a feed funnel, and a nitrogen inlet, and the apparatus was purged with nitrogen. CH2Cl2 (37.5L) and silica gel (15,000g) were loaded onto the column and thoroughly mixed and eluted to the surface of the silica gel. Sea sand (4000g) and magnesium sulfate (6000g) were sequentially packed into the column. Crude 3-(2,6-difluoro-3,5-dimethoxy-phenyl)-1-ethyl-N,N-dimethyl-2-oxo-1,2,3,4-tetrahydro-7H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidine-7-sulfonamide (6118g) and CH2Cl2 (22.5L) were thoroughly mixed until a solution was obtained, which was then packed into the column. The elution rate was found to be too slow, so the magnesium sulfate and solution were removed from the column and filtered. The filter cake was washed with CH2Cl2 (20 L) and the filtrate was loaded into a column. The container was rinsed with CH2Cl2 (2.5 L) and the rinse was loaded into the column. The column was eluted with 5% EtOAc / CH2Cl2 (prepared from 9.4 L of EtOAc and 178.1 L of CH2Cl2, respectively). The desired fraction was partially concentrated under vacuum at about 45 °C (using two rotary evaporators for convenience) to a target total volume of 24 L remaining (approximately 4 L per kg of 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-N,N-dimethyl-2-oxo-1,2,3,4-tetrahydro-7H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidine-7-sulfonamide). The distillation residue (24 L) was loaded into a 100 L reactor and the temperature was adjusted to about 28 °C. Heptane (54 L) was added and the reaction mixture was stirred at about 24 °C for 1 hour. The reaction mixture was filtered and the filter cake was washed with heptane (24 L). The product was air-dried on a filter for about 3 hours to provide product (5382 g). 21 H 23 LCMS calculated value of F2N5O5S [M+H] + : 495.5; Experimental value: 495.1. 1 H NMR (400MHz, DMSO-d6) δ8.12(s,1H),7.72(s,1H),7.07(t,1H),6.89(s,1H),4.83(s,2H),4.14(t,2H),3.91(s,6H),2.96(s,6H),1.35(t,3H).

[0374] Step 2. Synthesis of 7-(2,6-difluoro-3,5-dimethoxy-phenyl)-9-ethyl-2-formyl-8-oxo-6,7,8,9-tetrahydro-3,4,7,9-tetraaza-cyclopentadieno[a]naphthalene-3-sulfonic acid dimethylamide

[0375]

[0376] A 22L and a 200L glass reactor were assembled with an elevated stirred condenser, thermocouples, a feed funnel, and a nitrogen inlet, and all equipment was purged with nitrogen. THF (2.38L) and N,N-diisopropylamine (0.82L) were charged into the 22L glass reactor, and the mixture was cooled to -72°C. A 2.5M solution (2.13L) of n-BuLi in hexane was added, while maintaining the temperature at approximately -70°C. The reaction mixture was stirred at -71°C for approximately 7 minutes and then heated to -2°C over 3.5 hours to form a 1M LDA solution.

[0377] 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-N,N-dimethyl-2-oxo-1,2,3,4-tetrahydro-7H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidine-7-sulfonamide (1075 g gross weight, 93.07 wt%, 1001 g net weight) and THF (10.0 L) were loaded into the first rotary evaporator and rotated at about 63 °C for about 28 minutes without solvent collection until a solution was obtained. 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-N,N-dimethyl-2-oxo-1,2,3,4-tetrahydro-7H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidine-7-sulfonamide (1075 g gross weight, 93.07 wt%, 1001 g net weight) and THF (10.0 L) were loaded into a second rotary evaporator and rotated at about 61 °C for about 35 min without solvent collection until a solution was obtained. The two mixtures were concentrated under reduced pressure at about 50 °C. THF (10.0 L per rotary evaporator) was loaded into each rotary evaporator, and the two mixtures were concentrated under reduced pressure at about 50 °C. THF (10.0 L) was loaded into the first rotary evaporator, and the mixture was rotated at about 64 °C for 14 min without solvent collection until a solution was obtained. THF (10.0 L) was charged into a second rotary evaporator and the mixture was rotated at approximately 64 °C for 14 minutes without solvent collection until a solution was obtained. The percentage of residual CH₂Cl₂ (PCT) obtained by GC was satisfactory. Both solutions were transferred to a 200 L reactor, using THF (26 L) to aid the transfer. The reaction mixture was cooled to -65 °C. 1 M LDA solution (4.84 L) was charged over approximately 1 hour while maintaining the temperature at approximately -64 °C, and the reaction mixture was stirred at -65 °C for 2.25 hours. DMF (1.56 L) was charged over 30 minutes while maintaining the temperature at approximately -65 °C. The reaction mixture was stirred at -64 °C for 31 minutes and then heated to -12 °C over approximately 2 hours. An aqueous solution of ammonium chloride was prepared separately by thoroughly mixing ammonium chloride (160 g) with drinking water (1.6 L). An aqueous solution of ammonium chloride was added over a period of 17 minutes while maintaining the temperature at approximately -5°C, and the reaction mixture was heated to 19°C over a period of approximately 7.5 hours. The reaction mixture was then partially concentrated under vacuum at approximately 45°C (using two rotary evaporators for convenience). A total volume of 36 L was collected by distillation (approximately 18 L was collected per kilogram of 3-(2,6-difluoro-3,5-dimethoxy-phenyl)-1-ethyl-N,N-dimethyl-2-oxo-1,2,3,4-tetrahydro-7H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidine-7-sulfonamide).The distillation residue was loaded into a 200 L reactor and the temperature was adjusted to approximately 23 °C. The reaction mixture was stirred at 23 °C for approximately 2 h. Drinking water (20.0 L) was added over approximately 1 h, and the reaction mixture was stirred at approximately 23 °C. The reaction mixture was filtered, and the filter cake was washed twice with drinking water (10.0 L each time). The product was air-dried on a filter for approximately 3.5 h to provide a crude product (2028 g). The crude product was slurried in MTBE at 46–53 °C for 1 h, then cooled to room temperature, filtered, and washed with MTBE to give 7-(2,6-difluoro-3,5-dimethoxy-phenyl)-9-ethyl-2-formyl-8-oxo-6,7,8,9-tetrahydro-3,4,7,9-tetraaza-cyclopentadieno[a]naphthalene-3-sulfonic acid dimethylamide: C. 22 H 23 LCMS calculated value of F2N5O6S [M+H] + : 524.13; Experimental value: 524.80. 1 H NMR (400MHz, DMSO-d6) δ10.24(s,1H),8.31(s,1H),7.50(s,1H),7.05(m,1H),4.86(s,2H),4.17(m,2H),3.86(s,6H),3.04(s,6H),1.29(m,3H).

[0378] Step 3. Synthesis of 7-(2,6-difluoro-3,5-dimethoxy-phenyl)-9-ethyl-2-morpholin-4-ylmethyl-8-oxo-6,7,8,9-tetrahydro-3,4,7,9-tetraaza-cyclopentadieno[a]naphthalene-3-sulfonic acid dimethylamide

[0379] A 200L glass reactor was assembled with an elevated stirred condenser, thermocouples, a feed funnel, and a nitrogen inlet, and the apparatus was purged with nitrogen. Methane chloride (20.0L) and 7-(2,6-difluoro-3,5-dimethoxy-phenyl)-9-ethyl-2-formyl-8-oxo-6,7,8,9-tetrahydro-3,4,7,9-tetraaza-cyclopentadieno[a]naphthalene-3-sulfonic acid dimethylamide (2000g) were added to the reactor and the mixture was stirred at 18°C ​​until a solution was obtained. Morpholine (1.7L) was added to the reaction mixture. Acetic acid (2.0L) was added over 34 minutes while maintaining the temperature at approximately 32°C. The reaction mixture was stirred at approximately 27°C for 4 hours. Sodium triacetoxyborohydride (1620g) was added over 40 minutes while maintaining the temperature at approximately 28°C. The reaction mixture was stirred at approximately 24°C for 2.5 hours. Separately, a solution was prepared by thoroughly mixing sodium bicarbonate (2800 g) with drinking water (40.0 L) until a solution was obtained. The solution was added over 36 minutes while maintaining the temperature at approximately 19°C until a pH of 8–9 was obtained. The reaction mixture was stirred at approximately 18°C ​​for 30 minutes. The phases were separated. The organic phase was extracted three times with methane chloride (10.0 L each time). The combined organic phases were washed with drinking water (20.0 L). Separately, a solution was prepared by thoroughly mixing sodium chloride (2001 g) with drinking water (20.0 L) until a solution was obtained. The organic phase was washed with sodium chloride solution and dried over MgSO4 (600 g). The reaction mixture was filtered and the filter cake was washed with methane chloride (6.0 L). The combined filtrate and washings were concentrated under reduced pressure at 38°C to provide crude 7-(2,6-difluoro-3,5-dimethoxy-phenyl)-9-ethyl-2-morpholin-4-ylmethyl-8-oxo-6,7,8,9-tetrahydro-3,4,7,9-tetraaza-cyclopentadieno[a]naphthalene-3-sulfonic acid dimethylamide (2200 g). Chromatography using 0-40% EtOAc / DCM yielded 7-(2,6-difluoro-3,5-dimethoxy-phenyl)-9-ethyl-2-morpholin-4-ylmethyl-8-oxo-6,7,8,9-tetrahydro-3,4,7,9-tetraaza-cyclopentadieno[a]naphthalene-3-sulfonic acid dimethylamide: C 26 H 32 LCMS calculated value of F2N6O6S [M+H] + : 595.21; Experimental value: 474.2. 1H NMR(400MHz,DMSO-d6)δ8.10(s,1H),7.05(t,1H),6.81(s,1H),4.80(s,2H),4.12(m ,2H),3.91(s,6H),3.79(s,2H),3.57(m,4H),3.10(s,6H),2.50(m,4H),1.35(m,3H).

[0380] Example 7.3 Synthesis of 3-(2,6-difluoro-3-hydroxy-5-methoxyphenyl)-1-ethyl-8-(morpholinylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (compound 2)

[0381]

[0382] A mixture of 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-N,N-dimethyl-8-(morpholin-4-ylmethyl)-2-oxo-1,2,3,4-tetrahydro-7H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidine-7-sulfonamide (44.88 g, 75.48 mmol) in 1,4-dioxane (400 mL, 4000 mmol) was stirred under N2 while a mixture of 12.0 M aqueous hydrogen chloride solution (1260 mL, 1510 mmol) and water (1260 mL, 6990 mmol) was added via another funnel (internal temperature reached 35 °C). The resulting solution was heated at 80 °C for 18 h. LC-MS indicated no residue of the starting material. The reaction mixture was cooled to room temperature, the organic solvent was concentrated, and the resulting aqueous HCl solution was diluted with 200 mL of 2N HCl solution. The resulting aqueous solution was extracted with DCM (3 x 80 mL), and the combined DCM phases were further extracted with 6N HCl (80 mL). The combined HCl aqueous solution was stirred and cooled to 0-5°C with ice water. The acidic solution was neutralized to pH > 12 by dropwise addition of 25% NaOH (approximately 200 mL). The resulting solid (the unreacted, unprotected starting material or compound 1) was filtered and washed with water (3 x 250 mL). The alkaline aqueous solution was acidified with aqueous HCl to pH approximately 6 and extracted with DCM (3 x 30 mL). The combined DCM solutions were dried over Na₂SO₄ and concentrated. The crude material was then slurried in MTBE, filtered, washed with MTBE, and dried overnight in a vacuum oven at 50°C to obtain 3-(2,6-difluoro-3-hydroxy-5-methoxyphenyl)-1-ethyl-8-(morpholinylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (277 mg): C23 H 26 LCMS calculated value of F2N5O4 [M+H] + : 474.47; Experimental value: 474.2. 1 H NMR(400MHz,DMSO-d6)δ11.40(s,1H),9.95(s,1H),7.99(s,1H),6.78(m,1H),6.42(s ,1H),4.76(s,2H),4.09(m,2H),3.80(s,3H),3.60(m,6H),2.41(m,4H),1.28(m,3H).

[0383] Example 8. Synthesis of 3-(2,6-difluoro-3,5-bis(methoxy-d3)phenyl)-1-ethyl-8-(morpholinylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (compound 6)

[0384]

[0385] Step 1. Synthesis of 3-(2,6-difluoro-3,5-dihydroxyphenyl)-1-ethyl-8-(morpholinylmethyl)-7-(phenylsulfonyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one

[0386] A solution of 2.0 g (3.2 mmol) of 3-(2,6-difluoro-3,5-dimethoxyphenyl)-1-ethyl-8-(morpholinylmethyl)-7-(phenylsulfonyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (2.0 g, 3.2 mmol) in 25.0 mL of DCM was stirred at -78 °C while BBr3 (3.10 mL, 10.0 equivalent) (pure) was added dropwise. After 30 min, the dry ice bath was removed and the reaction mixture was slowly heated to room temperature. After 2 h, HPLC indicated the absence of starting material. The reaction mixture was then cooled to 0 °C, carefully treated with 20 mL of ice water, and stirred for 30 min. The resulting solid was filtered, washed with water, and dried overnight on a funnel. The crude material was treated with 10 mL of 20% MeOH DCM solution and stirred for 20 min, then filtered and dried to obtain a white solid 3-(2,6-difluoro-3,5-dihydroxyphenyl)-1-ethyl-8-(morpholinylmethyl)-7-(phenylsulfonyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (1.208 g, 61% yield): C 28 H 28LCMS calculated value of F2N5O6S [M+H] + : 600.17; Experimental value: 600.4. The material was used in the next step without further purification.

[0387] Step 2.3 Synthesis of (2,6-difluoro-3,5-bis(methoxy-d3)phenyl)-1-ethyl-8-(morpholinylmethyl)-7-(phenylsulfonyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one

[0388] A solution of DMF (12.4 mL, 160 mmol) containing 3-(2,6-difluoro-3,5-dihydroxyphenyl)-1-ethyl-8-(morpholinylmethyl)-7-(phenylsulfonyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (1.20 g, 2.00 mmol) and NaH (60% dispersion in mineral oil, 216 mg, 9.01 mmol) was stirred at room temperature under N2 for 15 min (all dissolved in the solution), and then MeI-D was added dropwise. 3 (0.263 mL, 4.20 mmol). After stirring at room temperature for 1 h, the reaction mixture was cooled to 0 °C, 30 mL of ice water was added, and the resulting solid was stirred for 30 min. The resulting solid was filtered, washed with water, and dried. The crude product was purified by Biotage chromatography using 0–6% MeOH / DCM to give 3-(2,6-difluoro-3,5-bis(methoxy-d3)phenyl)-1-ethyl-8-(morpholinylmethyl)-7-(phenylsulfonyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (950 mg, 75% yield) as a white solid. 30 H 26 LCMS calculated value of D6F2N5O6S [M+H] + : 634.23; Experimental value: 634.5. 1 H NMR(400MHz,DMSO-d6)δ8.41(m,2H),8.07(s,1H),7.70(m,1H),7.63(m,2H),7.05(m,1H),6 .89(s,1H),4.76(s,2H),4.09(m,2H),3.93(s,2H),3.60(m,4H),2.50(m,4H),1.28(m,3H).

[0389] Step 3.3 Synthesis of (2,6-difluoro-3,5-bis(methoxy-d3)phenyl)-1-ethyl-8-(morpholinylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one

[0390] Add 1,4-dioxane (10.0 mL) containing 3-(2,6-difluoro-3,5-bis(methoxy-d3)phenyl)-1-ethyl-8-(morpholinylmethyl)-7-(phenylsulfonyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (1000 mg, 1.578 mmol) and 1N sodium hydroxide aqueous solution (6312 μl, 6.31 mmol) to a 25 mL flask. Heat the solution to 74 °C (internal temperature) for 15 hours. LC-MS indicates the reaction is complete. Cool the clear, pale yellow solution to room temperature (as the mixture cools, solids begin to precipitate from the solution), giving a grayish-white suspension. Add water (10.0 mL) at 20–25 °C and stir the resulting solid for 30 min. The solid was filtered, washed six times with water, and the pH of the final wash was tested (pH = approximately 7). The crude product (0.70 g) was analyzed by ¹H NMR, indicating the presence of some dioxane.

[0391] DCM (3.88 mL, 60.3 mmol) containing crude 3-(2,6-difluoro-3,5-bis(methoxy-d3)phenyl)-1-ethyl-8-(morpholinylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one (0.70 g, 1.418 mmol) was heated to 22–36 °C and stirred to obtain a clear solution. After about 5 min, MTBE (0.693 mL, 5.82 mmol) was added. After stirring for about 1 h, a precipitate formed, and the mixture was stirred again for 30 min. The mixture was filtered, washed with MTBE, washed with heptane, and dried in a vacuum oven at 50 °C under N2 to give the product (0.63 g, 81% yield, 96% HPLC purity). The crude solid was purified by Biotage chromatography using 0-10% MeOH / DCM. The fractions were combined, concentrated, and dried in a vacuum oven at 50°C to obtain 3-(2,6-difluoro-3,5-bis(methoxy-d3)phenyl)-1-ethyl-8-(morpholinylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one with a purity of 99.5% HPLC. 24 H 22 LCMS calculated value of D6F2N5O4 [M+H] +: 494.24; Experimental value: 494.2. 1 HNMR (400MHz, DMSO-d6) δ11.75(S,1H),7.99(s,1H),7.05(m,1H),6.89(s,1H),4.76(s,2H),4.09(m,2H),3.60(m,6H),2.50(m,4H),1.28(m,3H).

[0392] Compound 6 can also be prepared by replacing 2,6-difluoro-3,5-bis(methoxy-d3)aniline with 2,6-difluoro-3,5-dimethoxyaniline according to Example 1. 2,6-difluoro-3,5-bis(methoxy-d3)aniline can also be prepared by replacing sodium methoxide with sodium methoxide according to step 3 of Example 4.

[0393] Example A

[0394] FGFR enzyme analysis

[0395] The inhibitory properties of the illustrated compounds were determined in an enzyme discontinuous assay, which uses FRET measurements to measure peptide phosphorylation and detect product formation. Inhibitors were serially diluted in DMSO, and 0.2 μL volumes were transferred to the wells of a 384-well plate. For FGFR isotypes (-1, -2, -3 wild-type and mutant isotype, -4) including phosphorylated and non-phosphorylated proteins, 5 μL of the enzyme diluted in assay buffer (50 mM HEPES, 10 mM MgCl2, 1 mM EGTA, 0.01% Tween-20, 5 mM DTT, pH 7.5) was added to each well and pre-incubated with the inhibitor at ambient temperature for 5–15 minutes. Appropriate controls (enzyme blank and enzyme without inhibitor) were included on the plate. The reaction was initiated by adding 5 μL of assay buffer per well containing a biotinylated EQEDEPEGDYFEWLE (SEQ ID NO.1) peptide substrate and ATP. The peptide substrate was reacted at a concentration of 500 nM (10 μL / well), while the ATP concentration was maintained at or below the ATP Km for each FGFR isoform. The ATP Km value was pre-determined for each FGFR isoform in individual experimental series. The reaction plates were incubated at 25 °C for 1 hour and the reaction was terminated by adding 5 μL of quench solution per well (50 mM Tris, 150 mM NaCl, 0.5 mg / mL BSA, pH 7.8; 45 mM EDTA, 600 nM staurosporin, 3.75 nM Eu-antibody PY20, and 180 nM APC-streptavidin in Perkin Elmer Lance reagent). The plates were allowed to equilibrate at ambient temperature for approximately 10 minutes before scanning on a PheraStar plate reader (BMG Labtech).

[0396] Data were analyzed using GraphPad Prism or XLfit. The IC was obtained by fitting the data to a four-parameter logistic equation that produces an S-shaped dose-response curve with a variable Hill coefficient. 50 Value. Prism equation: Y = minimum value + (maximum value - minimum value) / (1 + 10^(LogIC)) 50 -X)*Hill slope);

[0397] The XLfit equation is: Y = (A + ((BA) / (1 + ((X / C)^D)))), where X is the logarithm of the inhibitor concentration and Y is the reaction.

[0398] FGFR inhibition data for various compounds disclosed herein are shown in Table 1 below. Symbol: “+” indicates IC50. 50 Less than 10nM; “++” indicates IC 50 Greater than or equal to 10 nM but less than 100 nM; "+++" indicates IC 50 Greater than or equal to 100 nM but less than 500 nM; and “++++” indicates IC 50 Greater than or equal to 500 nM but less than 1000 nM.

[0399] Table 1. FGFR suppression data

[0400]

[0401]

[0402] Example B: KATOIII whole blood pFGFR2α ELISA analysis

[0403] To measure tyrosine-phosphorylated fibroblast factor receptor 2α (FGFR2α) in whole blood supplemented with KATO III cells, KATO III cells were purchased from ATCC and maintained in Iscove's medium (Gibco / Life Technologies) containing 20% ​​FBS. To measure the inhibition of FGFR2α activity by the test compounds, Iscove's medium (0.2% FBS) was used at 5 x 10⁻⁶ cells / mL. 6 Cells were resuspended at 1 cell / ml. Then, with or without a specified concentration range of the test compound and 300 μL of human heparinized whole blood (Biological Specialty Corp, Colmar PA), 50 μL of cells were added to a 2 ml 96-well polypropylene analytical block (Costar). After incubation at 37°C for 4 hours, erythrocytes were lysed with Qiagen EL buffer, and the lysate was resuspended in a lysis buffer (Cell Signaling) containing a standard protease inhibitor mixture (Calbiochem / EMD) and PMSF (Sigma) for 30 minutes on ice. The lysate was transferred to a standard V-bottom propylene tissue culture plate and frozen overnight at -80°C. Samples were tested in an R&DSystems DuoSet IC human phosphate-FGF R2α ELISA plate, and measurements were taken using a SpectraMax M5 microplate set to 450 nm with wavelength correction at 540 nm. IC50 analysis was performed using GraphPad Prism 5.0 software to fit a curve of the percentage of inhibition versus the logarithm of the inhibitor concentration. 50 Measurement.

[0404] KATOIII whole blood pFGFR2α ELISA IC 50 The data is shown in Table 2 below. Symbol: "+" indicates IC 50 Less than 50nM; “++” indicates IC 50 Greater than or equal to 50 nM but less than 250 nM; "++" indicates IC 50 Greater than or equal to 250 nM but less than 500 nM; “++++” indicates IC 50 Greater than or equal to 500 nM but less than 1000 nM; and “+++++” indicates IC 50 Greater than or equal to 1000 nM but less than 3000 nM.

[0405] Table 2. KATOIII Whole Blood pFGFR2α ELISA Data

[0406]

[0407] Example C: Determination of osmotic pressure and P-gp-mediated transport in Caco-2 cells

[0408] Caco-2 cells were grown in DMEM medium at a seeding density of 14,000 cells / well in 96-well Transwell plates. To determine permeability in the absorption direction (AB), HBSS containing the test compound was added to the donor compartment (top side), and HBSS containing 4% BSA was added to the receiver compartment (base side). To determine whether the compound was a P-gp substrate, permeability was measured in both the AB and BA directions in the absence of 4% BSA (bidirectional transport assay). Digoxin and cyclosporine A were included as positive controls for P-gp substrate and inhibitor, respectively, to ensure the functionality of P-gp in the bidirectional transport assay. Various concentrations of the test compound were added to the donor compartment (AB transport is top side and BA transport is base side), while HBSS solution was added to the receiver compartment (AB transport is base side and BA transport is top side). In the AB permeability study, the donor volume was 0.075 mL and the receiver volume was 0.25 mL. In the osmotic studies along the BA side, the donor volume was 0.25 mL and the receiver volume was 0.075 mL. Incubation was performed at 37°C for 120 minutes. Transmembrane resistance (TEER) was measured before and after the 120-minute incubation to ensure the integrity of the cell monolayer. At the end of the incubation period, the sample was removed from both the donor and receiver sides and mixed with acetonitrile for protein precipitation. The supernatant was collected after centrifugation for analysis using LC-MS / MS. The osmotic coefficient (Papp) value for the Caco-2 studies was determined using the following equation:

[0409] Papp (cm / s) = (F*VD) / (SA*MD)

[0410] Where the flow rate (F, mass / time) is calculated from the slope of the cumulative amount of the compound of interest on the receiver side, SA is the surface area of ​​the cell membrane, VD is the donor volume, and MD is the initial amount of solution in the donor chamber.

[0411] The outflow ratio of Caco-2 was calculated as the ratio of Papp measured in the BA direction to Papp in the AB direction.

[0412] The permeability data of the various compounds disclosed herein in Caco-2 cells are shown in Table 3 below.

[0413] Table 3. Osmolysis in Caco-2 cells

[0414]

[0415] Data on P-gp-mediated transport of various compounds disclosed herein in Caco-2 cells are shown in Tables 4 and 5 below.

[0416] Table 4. Bidirectional transport of compound 2 across the Caco-2 monolayer

[0417]

[0418] NA = Not applicable; CSA = Cyclosporin A; *N = 3

[0419] Table 5. Bidirectional transport of compound 1 across the Caco-2 monolayer

[0420]

[0421]

[0422] NA = Not applicable; CSA = Cyclosporin A; *N = 3-6

[0423] Compound 1 showed an efflux ratio of 0.64 at 30 μM. In contrast, compound 2 showed an efflux ratio of 19 at 30 μM. This indicates that compound 2 is a more efficient substrate for Pgp transport compared to compound 1, and compound 2 is not well absorbed into the bloodstream. Compound 2 is a candidate for intravenous administration or hepatic artery infusion to treat diseases and conditions such as cholangiocarcinoma.

[0424] Furthermore, compound 1 had a permeability of 11 μm in Caco-2 cells, while compound 2 had a permeability of 0.5 μm. Given the small differences in their structural composition, the difference in permeability and efflux ratio between compounds 1 and 2 was quite unexpected.

[0425] Example D: Study on the absorption, metabolism and excretion of compound 1 in humans

[0426] An open-label study was conducted to evaluate the mass balance, pharmacokinetics, and metabolite morphology of a single oral dose of compound 1 [14C].

[0427]

[0428] Seven male subjects each received a single oral dose of 11 mg of compound 1 tablet and a solution of [14C] compound 1 (approximately 250 μCi) after an overnight fast. Blood / plasma, urine, and stool samples were collected from the participants 4 to 10 days after administration.

[0429] Human elimination criteria include recovering at least 90% of the applied radiation dose; and recovering less than or equal to 1% of the applied radiation dose in excrement (combined urine and feces) from two consecutive 24-hour urine and fecal collection samples.

[0430] 12.6% of the dose was recovered in urine and 82.4% in feces. The total radioactivity recovery rate in urine and feces was 95.1% over the 240-hour study. Rapid absorption was observed, with peak total radioactivity and plasma compound 1 concentrations observed approximately 2.0 hours post-dose.

[0431] Figure 1 The mean cumulative radiation dose percentage recovered in urine and feces at specified intervals after a single 13-mg (250 μCi) oral dose of compound 1 [14C] is shown.

[0432] Figure 2 The mean radioactivity (nM equivalent) in blood or plasma and the concentration of compound 1 in plasma (nM) after a single oral dose of approximately 13 mg [14C] compound 1 in a fasting healthy male volunteer are shown. Total plasma radioactivity and compound 1 have a half-life of approximately ten hours. The ratio of plasma compound 1 concentration to total plasma radioactivity is approximately 0.7, indicating that compound 1 is a major circulating component in plasma. The blood-to-plasma radioactivity ratio (approximately 0.8) indicates a low correlation between radioactivity and blood cells.

[0433] Four minor circulating metabolites (<10% of the compound-related material) were observed, as shown below:

[0434]

[0435] Compound 2-1 is a glucuronide derivative of compound 2. Compound 2-2 is a sulfonic acid derivative of compound 2. Compound 1-1 is a ketone derivative of compound 1. The rings surrounding certain parts of the chemical structure indicate the selection of the glucuronide, sulfonic acid, and ketone groups.

[0436] The percentage of each compound isolated from the blood / plasma sample relative to the administered dose is shown in the table below:

[0437]

[0438] Figure 3 The mass spectrum of the human circulating metabolite of compound 1 is shown.

[0439] 12.6% of the dose was recovered in the urine. Two metabolites of compound 1 were isolated from the urine, as shown below:

[0440]

[0441] The rings surrounding certain parts of the chemical structure indicate the choice of connecting glucuronic acid and sulfonic acid groups.

[0442] The percentage of each compound isolated from the urine sample relative to the administered dose is shown in the table below:

[0443]

[0444] Figure 4 The mass spectrum of the metabolite of compound 1 isolated from urine is shown.

[0445] 82.4% of the dose was recovered in feces. The following metabolites of compound 1 were isolated from feces:

[0446]

[0447] Compound 4-1 is a ketone derivative of compound 4. Compounds 2-3 are ketone derivatives of compound 2. Compounds 1-2 are dihydroxylated derivatives of compound 1. The rings surrounding certain parts of the chemical structure indicate the choice of which hydroxyl and ketone groups are attached.

[0448] The percentages of each compound isolated from the fecal sample relative to the administered dose are shown in the table below:

[0449]

[0450] Figure 5 The mass spectrum of the metabolite of compound 1 isolated from feces is shown.

[0451] From the foregoing description, various modifications to the invention, other than those described herein, will be apparent to those skilled in the art. Such modifications are also intended to fall within the scope of the appended claims. Every reference listed in this application, including all patents, patent applications, and publications, is incorporated herein by reference in its entirety.

Claims

1. A compound, said compound being: 3-(2,6-difluoro-3-hydroxy-5-methoxyphenyl)-1-ethyl-8-(morpholinylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one, Or its pharmaceutically acceptable salt.

2. The compound of claim 1, wherein the compound is 3-(2,6-difluoro-3-hydroxy-5-methoxyphenyl)-1-ethyl-8-(morpholinylmethyl)-1,3,4,7-tetrahydro-2H-pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-2-one.

3. A composition comprising the compound as claimed in claim 1 or 2 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.

4. The composition of claim 3, wherein the composition is suitable for oral administration.

5. The composition of claim 3, wherein the composition is suitable for intravenous administration.

6. The composition of claim 3, wherein the composition is suitable for arterial administration.

7. The composition of claim 6, wherein the arterial administration is a hepatic artery infusion.

8. Use of the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof or the composition of any one of claims 3-7 in the preparation of a medicament for inhibiting FGFR enzymes.

9. Use of the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof or a composition of any one of claims 3-7 in the preparation of a medicament for FGFR-mediated cancer.

10. The use as described in claim 9, wherein the cancer is selected from bladder cancer, breast cancer, cervical cancer, colorectal cancer, endometrial cancer, gastric cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, ovarian cancer, prostate cancer, esophageal cancer, gallbladder cancer, pancreatic cancer, thyroid cancer, skin cancer, leukemia, multiple myeloma, chronic lymphocytic lymphoma, B-cell lymphoma, Hodgkin's or non-Hodgkin's lymphoma, Waldenström's macroglobulinemia, pilocellular lymphoma, Burkitt's lymphoma, glioblastoma, melanoma, and rhabdomyosarcoma.

11. The use as claimed in claim 9, wherein the cancer is selected from adult T-cell leukemia and acute myeloid leukemia.

12. Use of the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof or the composition of any one of claims 3-7 in the preparation of a medicament for FGFR-mediated myeloproliferative disorders.

13. The use as described in claim 12, wherein the myeloproliferative disorder is selected from polycythemia vera, spontaneous thrombocytosis, and primary myelofibrosis.

14. Use of the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof or a composition of any one of claims 3-7 in the preparation of a medicament for treating FGFR-mediated bone or chondrocyte disorders.

15. The use as claimed in claim 14, wherein the skeletal or chondrocyte disorder is selected from dwarfism, lethal dysplasia (TD), Aberle syndrome, Krusen syndrome, Jackson-Wes syndrome, Bill Stevenson skin swirl syndrome, Pfaffifol syndrome, and craniosynostosis syndrome.

16. The use as claimed in claim 14, wherein the bone or chondrocyte disease is selected from chondrodysplasia and reduced cartilage production.

17. Use of the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof or a composition of any one of claims 3-7 in the preparation of a medicament for FGFR-mediated hypophosphatemia.

18. The use as claimed in claim 17, wherein the hypophosphatemia condition is X-linked hypophosphatemic rickets, autosomal recessive hypophosphatemic rickets, autosomal dominant hypophosphatemic rickets, or tumor-induced osteomalacia.

19. Use of the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof or a composition of any one of claims 3-7 in the preparation of a medicament for FGFR-mediated cholangiocarcinoma.

20. The use as claimed in claim 19, wherein the cholangiocarcinoma is advanced or metastatic cholangiocarcinoma.

21. Use of the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof or the composition of any one of claims 3-7 in the preparation of a medicament for FGFR-mediated myeloma / lymphoma.

22. The use as claimed in claim 21, wherein the bone marrow / lymphoma is 8p11 myeloproliferative syndrome.

23. The use as claimed in claim 21, wherein the bone marrow / lymphoma is associated with eosinophilia.

24. A method for preparing compound 2 having the following formula or a salt thereof: Compound 2, The method includes: a) Make compound F1 have the following formula: Compound F1, It reacts with an amino protecting agent to provide compound F2 having the following formula: Compound F2, or its salt, wherein P 1 It is an amino protecting group; b) React compound F2 with DMF in the presence of B1, where B1 is a base, to provide compound F3 having the following formula: Compound F3, or its salt; c) React compound F3 with morpholine in the presence of RA1, where RA1 is a reducing agent, to provide compound F4 having the following formula: Compound F4, or its salt; as well as d) Deprotect compound F4 to provide compound 2 or its salt.

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