# Patent

## Title
Substituted azaindole compounds, salts, pharmaceutical compositions thereof and methods of use

## Abstract

The present invention provides substituted azaindole prodrugs, methods of making said prodrugs, pharmaceutical compositions of said prodrugs and methods of using said prodrugs and pharmaceutical compositions thereof to treat or prevent diseases or disorders such as cancer.

## Background
CROSS-REFERENCE TO RELATED APPLICATION
This application claims the benefit of U.S. Provisional Application Ser. No. 61/693,335, filed on Aug. 27, 2012, which is hereby incorporated by reference in its entirety.


FIELD OF THE INVENTION
The present invention relates generally to substituted azaindole prodrugs, methods of making said prodrugs, pharmaceutical compositions of said prodrugs and methods of using said prodrugs and pharmaceutical compositions of using said prodrugs to treat or prevent diseases or disorders such as cancer.
BACKGROUND OF THE INVENTION
The information provided herein is intended solely to assist the understanding of the reader. None of the information provided nor references cited is admitted to be prior art to the present invention. Each of the references cited herein is incorporated herein by reference in its entirety.
Kinases are a large family of proteins that have now become firmly established as a major class of drug targets. The sequencing of the Human Genome has led to the identification of over 500 protein kinases encoded within it: the Human Kinome. These protein kinases have been grouped into a total of seven families, based on their structures. The 388 serine/threonine kinases fall into five families: AGC, CAMK, CMGC, CK1, and STE. The 90 tyrosine kinases fall into two families: the TK (tyrosine kinase) family of 58 kinases and the TKL (tyrosine kinase-like) family of 32 kinases. There are 40 atypical kinases that are highly structurally distinct from the remainder. The TK family is a source forvalidated drug targets, such as epidermal growth factor receptor (EGFR), vascular endothelial growth factor receptor (VEGFR), and platelet-derived growth factor receptor (PDGFR). Recent clinical data provided proof-of-concept for selective B-Raf inhibitors in the treatment of B-RafV600E mutant melanoma, which led to the approved clinical use of vemurafenib for the treatment of late stage metastatic melanoma.
The regulation of cancer cell growth is often modulated by multiple kinases acting both in independent pathways and in concerted signaling cascades. Thus, the Ras/Raf/MEK/ERK mitogen-activated protein kinase (MAPK) pathway mediates cellular responses to different growth signals and is frequently deregulated in cancer. The Raffamily consist of serine/threonine kinases A-Raf, B-Raf and C-Raf (Raf-1), which phosphorylate and activate MEK. Only B-Raf is frequently mutated in various cancers; and the most common B-Raf mutation involves a substitution of a glutamic acid residue to a valine moiety at codon 600 (BRAFV600E). Mutations in the B-Raf gene may lead to MAPK pathway amplification via constitutive activation of B-Raf kinase, even in the absence of any growth signals, resulting in increased malignancy.
A landmark study published in 2002 identified activating mutations in the B-Raf serine/threonine kinase gene BRAF in more than 50% of all melanomas. In addition to melanoma, activating BRAF mutations have also been identified in colorectal carcinomas (where it occurs in 40% of tumors with mismatch repair deficiency), 50% of thyroid papillary carcinomas and 30% of low-grade serous ovarian carcinomas. Although related isoforms of Raf, such as A-Raf and C-Raf, can activate other signaling pathways, it is believed that the transforming activity of B-Raf arises mainly through the stimulation of the MAPK pathway.
Melanomas are tumors derived from the transformation of melanocytes, a family of specialized pigment-producing cells. Melanomas are broadly divided into those arising from the skin (cutaneous), eyes (uveal) or mucous membranes (mucosal). Although melanoma is only the third most common form of skin cancer in people of European descent (after squamous cell carcinoma and basal cell carcinoma), it accounts for nearly all skin cancer deaths. Once disseminated to distant organs, melanoma becomes a considerable clinical problem associated with high rates of morbidity and mortality. Currently, median survival for stage IV melanoma is between 6 and 10 months, a rate that has changed little over the last 30 years.
Vemurafenib (ZELBORAF®; also known as PLX4032) is a small-molecule B-Raf kinase inhibitor for the treatment of cancers harboring activating BRAF mutations. The primary focus of vemurafenib use is in melanoma (>50% harbor activating BRAF mutations). The applications of vemurafenib with other solid tumors, such as colorectal carcinoma (>10% harbor BRAF mutations), are also under investigation. Purified kinase assays have demonstrated that vemurafenib and its related analogs are highly potent inhibitors of B-Raf activity, with 3-fold selectivity for the V600E mutation over the wild-type kinase.
In preclinical models, vemurafenib and its analogs inhibited the growth of B-RafV600E positive melanoma cell lines both in vitro and in vivo. In the initial stage of phase I clinical study, cohorts of patients with advanced solid tumors were treated with escalating doses of vemurafenib (ranging from 200 to 1,600 mg), administered twice-daily (BID) as oral capsules. The crystalline formulation yielded only modest drug exposures, so vemurafenib had to be reformulated as a micro-precipitated bulk powder (MBP), which can increase drug bioavailability.
Vemurafenib have low solubility in water and consequently showed a low dissolution rate and as a result exhibited poor bioavailability, especially when administered orally in crystalline form. Poor bioavailability can cause unpredictable absorption of the compound by the patient, leading to unpredictability in dose/therapy effects. Often food may affect the ability of the patient to absorb such poorly bioavailable compounds, and requiring dosing regimens to be adjusted accordingly. Also, an erratic absorbed drug commands a large safety margin. Moreover, such a drug will require higher doses to achieve sufficiently high systemic or target concentrations for efficacy, which may not be practical in some cases or may leadto adverse effects in others (Testa et al., Prodrugs: bridging pharmacodynamic/pharmacokineticgaps. Curr. Opin. Chem. Biol. 2009, 13, 1-7).
Prodrugs are analogues of active pharmaceutical ingredient that following administration are converted or metabolized to an active form of the drug in vivo. Prodrugs are used to modify one or more aspects of the pharmacokinetics of a drug in a manner that enhances the therapeutic efficacy of a drug. Prodrugs can provide possibilities to overcome various barriers to drug formulation and delivery such as poor aqueous solubility, chemical instability, insufficient oral absorption, rapid pre-systemic metabolism, and toxicity.
Clearly, a significant problem with oral administration of vemurafenib is the bioavailability at larger doses, which are required for the therapeutic efficacy of vemurafenib (approved daily dose is 960 mg BID in MBP formulation). The superior efficacy of vemurafenib against melanoma makes vemurafenib a particular compelling candidate for the preparation of its analogues that can be easily formulated to provide better oral bioavailability. Thus, there is a need for prodrugs of vemurafenib with demonstrated enhanced oral bioavailability. In particular, masking nitrogen in azaindole NH moiety and/or in aryl sulfonamide NH in vemurafenib facilitated absorption throughout gastrointestinal tract. These vemurafenib analogues can enhance the convenience (by reducing the dose and dosing frequency), efficacy, and lessen side effect profile of vemurafenib. In addition, vemurafenib analogues may also be able to be formulated in solution form for intravenous administration or in extensive release forms.
SUMMARY OF THE INVENTION
These and other needs are satisfied by the disclosure herein of substituted azaindole prodrugs, methods of making said prodrugs, pharmaceutical compositions of said prodrugs and methods of using said prodrugs and pharmaceutical compositions thereof to treat or prevent diseases or disorders such as cancer.
Specifically, it has been found that compounds disclosed herein, and pharmaceutically acceptable compositions thereof, are effective as prodrugs of vemurafenib for treating or preventing diseases or disorders such as cancer.
The first aspect of the invention provides a compound of Formula (I):




or a racemic mixture, a stereoisomer, a geometric isomer, a tautomer, a solvate, an N-oxide, a metabolite, or a pharmaceutically acceptable salt thereof, wherein each of X and Y is as defined herein.

In some embodiments, each of X and Y is independently H, —C(═O)R3, —C(═O)OR4, —C(R1R2)OC(═O)R3, —C(R1R2)OC(═O)OR4 or —C(R1R2)OP(═O)(OR4)(OR4a), with the proviso that when Y is H, X is not —C(═O)CH3 or H;
each R1 and R2 is independently H, D, C1-6alkyl, C1-6haloalkyl, C3-6cycloalkyl, —(C1-4 alkylene)—(C3-6cycloalkyl), C3-6heterocyclyl or —(C1-4alkylene)-(C3-6heterocyclyl); or R1 and R2, together with the carbon atom they are attached to, form an unsubstituted or substituted 3-8 membered carbocyclic or heterocyclic ring;
each R3 is independently H, D, C1-10alkyl, C1-10haloalkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6alkylene)-(C1-6aryl), 5-10 membered heteroaryl or —(C1-6alkylene)-(5-10 membered heteroaryl), wherein each of the C1-10alkyl, C1-10haloalkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl and —(C1-6alkylene)-(5-10 membered heteroaryl) is unsubstituted or optionally substituted with 1, 2, 3 or 4 substituents independently selected from D, F, Cl, Br, I, C1-6alkyl, C1-6haloalkyl, —OH, —NH2, C1-6alkoxy and C1-6alkylamino; and
each R4 and R4a is independently H, C1-10alkyl, C1-10haloalkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl or —(C1-6alkylene)-(5-10 membered heteroaryl), wherein each of the C1-10alkyl, C1-10haloalkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl and —(C1-6alkylene)-(5-10 membered heteroaryl) is unsubstituted or optionally substituted with 1, 2, 3 or 4 substituents independently selected from D, F, Cl, OH, NH2, oxo (═O), C1-6alkyl, C1-6haloalkyl, C1-6alkoxy and C1-6alkylamino.
In other embodiments, each R1 and R2 is independently H, D or C1-3alkyl.
In other embodiments, each R3 is independently C1-10alkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10 aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl or —(C1-6alkylene)-(5-10 membered heteroaryl), wherein each of the C1-10alkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8-cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl and —(C1-6alkylene)-(5-10 membered heteroaryl) is unsubstituted or optionally substituted with 1, 2, 3 or 4 substituents independently selected from D, F, —OH, —OMe, —NH2, —NHMe, —NMe2 and C1-3alkyl.
In other embodiments, each R4 and R4a is independently H, C1-10alkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl) or C6-10aryl, wherein each of the C1-10alkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl) and C6-10aryl is unsubstituted or optionally substituted with 1, 2, 3 or 4 substituents independently selected from D, F, OH, OMe, oxo (═O), NH2, —NHMe, —NMe2 and C1-3alkyl.
In other embodiments, each of X and Y is independently H or —C(R1R2)OP(═O)(OH)2, with the proviso that when Y is H, X is not H.
In other embodiment, each of X and Y is independently H, —C(═O)R3, —C(═O)OR4, —C(R1R2)OC(═O)R3 or —C(R1R2)OC(═O)OR4, with the proviso that when Y is H, X is not —C(═O)CH3 or H.
In other embodiments, the acyl moiety (—C(═O)R3) is derived from an α-amino acid or an optical isomer thereof by removing the OH group from the carboxy group of the α-amino acid.
In other embodiments, the α-amino acid is isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, alanine, asparagine, aspartate, glutamate, glutamine, proline, serine, para-tyrosine, arginine, histidine, cysteine, glycine, sarcosine, N,N-dimethyl glycine, homoserine, norvaline, norleucine, ornithine, homocysteine, homophenylalanine, phenylglycine, ortho-tyrosine, meta-tyrosine or hydroxyproline.
In other embodiments, the α-amino acid is isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, alanine, asparagine, aspartate, glutamate, glutamine, proline, serine, tyrosine, arginineor histidine, each of which has a S-configuration at the α-position.
In other embodiments, the pharmaceutically acceptable salt is an alkali metal salt, an alkaline earth metal salt, or an ammonium or N+(C1-4 alkyl)4 salt.
In other embodiments, the pharmaceutically acceptable salt is a sodium salt, a lithium salt, a potassium salt, a calcium salt, a magnesium salt; an ammonium salt, a quaternary ammonium salt or a combination thereof.
In other embodiments, the pharmaceutically acceptable salt is a salt of an inorganic acid, an organic acid or a combination thereof, wherein the inorganic acid or the organic acid is hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, malic acid, 2-hydroxypropanic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, glucuronic acid, galacturonic acid, citric acid, tartaric acid, aspartic acid, glutamic acid, benzoic acid, cinnamic acid, p-toluenesulfonic acid, benzenesulfonic acid, mthanesulfonic acid, ethanesulfonic acid, trifluoromthanesulfonic acid or a combination thereof.
In another aspect, provided herein is a pharmaceutical composition comprising a compound disclosed herein, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle or a combination thereof. In some embodiments, the compound is a prodrug of vemurafenib, a B-Raf kinase inhibitor.
In another aspect, provided herein is a composition comprising a compound that undergoes metabolic transformations in vivo to provide metabolites that are inhibitors of protein kinase. In some embodiments, the metabolite is vemurafenib, a B-Raf kinase inhibitor.
In some embodiments, the pharmaceutical composition disclosed herein further comprises a therapeutic agent selected from the group consisting of chemotherapeutic agents, anti-proliferative agents, agents for treating atherosclerosis, agents for treating lung fibrosis and combinations thereof.
In other embodiments, the therapeutic agent is chlorambucil, melphalan, cyclophosphamide, ifosfamide, busulfan, carmustine, lomustine, streptozocin, cisplatin, carboplatin, oxaliplatin, dacarbazine, temozolomide, procarbazine, methotrexate, fluorouracil, cytarabine, gemcitabine, mercaptopurine, fludarabine, vinblastine, vincristine, vinorelbine, paclitaxel, docetaxel, topotecan, irinotecan, etoposide, trabectedin, dactinomycin, doxorubicin, epirubicin, daunorubicin, mitoxantrone, bleomycin, mitomycin, ixabepilone, tamoxifen, flutamide, gonadorelin analogues, megestrol, prednisone, dexamethasone, methylprednisolone, thalidomide, interferon alfa, leucovorin, sirolimus, temsirolimus, everolimus, afatinib, alisertib, amuvatinib, apatinib, axitinib, bortezomib, bosutinib, brivanib, cabozantinib, cediranib, crenolanib, crizotinib, dabrafenib, dacomitinib, danusertib, dasatinib, dovitinib, erlotinib, foretinib, ganetespib, gefitinib, ibrutinib, icotinib, imatinib, iniparib, lapatinib, lenvatinib, linifanib, linsitinib, masitinib, momelotinib, motesanib, neratinib, nilotinib, niraparib, oprozomib, olaparib, pazopanib, pictilisib, ponatinib, quizartinib, regorafenib, rigosertib, rucaparib, ruxolitinib, saracatinib, saridegib, sorafenib, sunitinib, tasocitinib, telatinib, tivantinib, tivozanib, tofacitinib, trametinib, vandetanib, veliparib, vemurafenib, vismodegib, volasertib, alemtuzumab, bevacizumab, brentuximabvedotin, catumaxomab, cetuximab, denosumab, gemtuzumab, ipilimumab, nimotuzumab, ofatumumab, panitumumab, ramucirumab, rituximab, tositumomab, trastuzumab, or a combination thereof.
In another aspect, provided herein is a method of preventing, managing, treating or lessening the severity of a proliferative disorder in a patient by administering to the patient with the compound disclosed herein, or the pharmaceutical composition disclosed. In other embodiments, the method further comprises administering a therapeutic agent to the patient.
In another aspect, provided herein is the compound disclosed herein or the pharmaceutical composition disclosed herein for use in preventing, managing, treating or lessening the severity of a proliferative disorder in a patient.
In some embodiments, the proliferative disorder is metastatic cancer. In other embodiments, the proliferative disorder is colon cancer, gastric adenocarcinoma, bladder cancer, breast cancer, kidney cancer, liver cancer, lung cancer, melanoma, thyroid cancer, a cancer of the head and neck, prostate cancer, pancreatic cancer, a cancer of the CNS, glioblastoma, or a myeloproliferative disorder. In further embodiments, the proliferative disorder is atherosclerosis or lung fibrosis.
In another aspect, provided herein is a method of inhibiting or modulating the activity of a protein kinase in a biological sample comprising contacting a biological sample with the compound disclosed herein, or the pharmaceutical composition disclosed herein. In some embodiments, the protein kinase is B-Raf.
In another aspect, provided herein is a method of inhibiting protein tyrosine kinase, the method comprises contacting the kinase with the compound disclosed herein, or with the composition disclosed herein. In some embodiments, provided herein is a method of inhibiting B-Raf kinase signaling, the method comprises contacting the receptor with the compound disclosed herein, or with the pharmaceutical composition disclosed herein.
In other embodiments, inhibition of the activity of a receptor protein kinase, such as B-Raf signaling, can be in a cell or a multicellular organism. If in a multicellular organism, the method disclosed herein may comprise administering to the organism the compound disclosed herein, or the pharmaceutical composition disclosed herein. In some embodiments, the organism is a mammal; in other embodiments, the organism is a human. In still other embodiments, the method further comprises contacting the kinase with a therapeutic agent.
In another aspect, provided herein is a method of inhibiting proliferative activity of a cell, wherein the method comprises contacting the cell with an effective proliferative inhibiting amount of the compound disclosed herein or the pharmaceutical composition disclosed herein. In some embodiments, the method further comprises contacting the cell with an additional therapeutic agent.
In another aspect, provided herein is a method of treating a cell proliferative disease in a patient, wherein the method comprises administering to the patient in need of such treatment an effective therapeutic amount of the compound disclosed herein or the pharmaceutical composition disclose herein. In some embodiments, the method further comprises administering an additional therapeutic agent.
In another aspect, provided herein is a method of inhibiting tumor growth in a patient, the method comprises administering to the patient in need thereof treatment an effective therapeutic amount of the compound disclosed herein or the pharmaceutical composition disclose herein. In some embodiments, the method further comprises administering an additional therapeutic agent.
In another aspect, provided herein includes methods of preparing, methods of separating, and methods of purifying compounds of Formula (I).
The foregoing merely summarizes certain aspects disclosed herein and is not intended to be limiting in nature. These aspects and other aspects and embodiments are described more fully below.


DETAILED DESCRIPTION OF THE INVENTION
Definitions and General Terminology
Reference will now be made in detail to certain embodiments of the invention, examples of which are illustrated in the accompanying structures and formulas. The invention is intended to cover all alternatives, modifications, and equivalents which may be included within the scope of the present invention as defined by the claims. One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described herein. In the event that one or more of the incorporated literature, patents, and similar materials differs from or contradicts this application, including but not limited to defined terms, term usage, described techniques, or the like, this application controls.
As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, and the Handbook of Chemistry and Physics, 75th Ed. 1994. Additionally, general principles of organic chemistry are described in Sorrell et al., “Organic Chemistry”, University Science Books, Sausalito: 1999, and Smith et al., “March's Advanced Organic Chemistry”, John Wiley & Sons, New York: 2007, all of which are incorporated herein by reference.
As described herein, compounds disclosed herein may optionally be substituted with one or more substituents, such as those are illustrated below, or as exemplified by particular classes, subclasses, and species of the invention. It will be appreciated that the phrase “optionally substituted” is used interchangeably with the phrase “substituted”. In general, the term “substituted” refers to the replacement of one or more hydrogen radicals in a given structure with the radical of a specified substituent. Unless otherwise indicated, an optionally substituted group may have a substituent at each substitutable position of the group. When more than one position in a given structure can be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at each position. Examples of substituent include, but are not limited to, D, F, Cl, Br, I, oxo (═O), alkyl, haloalkyl, OH, NH2, alkoxy and alkylamino.
The term “alkyl” or “alkyl group” refers to a saturated linear or branched-chain monovalent hydrocarbon radical of one to twenty carbon atoms, wherein the alkyl radical may be optionally substituted independently with one or more substituents described below. Unless otherwise specified, alkyl groups contain 1-20 carbon atoms. In some embodiments, alkyl groups contain 1-10 carbon atoms. In another embodiment, alkyl groups contain 1-8 carbon atoms. In some embodiments, alkyl groups contain 1-6 carbon atoms. In another embodiment, alkyl groups contain 1-4 carbon atoms. In still some embodiments, alkyl groups contain 1-3 carbon atoms and in yet other embodiments, alkyl groups contain 1-2 carbon atoms.
Some non-limiting examples of alkyl groups include methyl (Me, —CH3), ethyl (Et, —CH2CH3), 1-propyl (n-Pr, n-propyl, —CH2CH2CH3), 2-propyl (i-Pr, i-propyl, —CH(CH3)2), 1-butyl (n-Bu, n-butyl, —CH2CH2CH2CH3), 2-methyl-1-propyl (i-Bu, i-butyl, —CH2CH(CH3)2), 2-butyl (s-Bu, s-butyl, —CH(CH3)CH2CH3), 2-methyl-2-propyl (t-Bu, t-butyl, —C(CH3)3), 1-pentyl (n-pentyl, —CH2CH2CH2CH2CH3), 2-pentyl (—CH(CH3)CH2CH2CH3), 3-pentyl (—CH(CH2CH3)2), 2-methyl-2-butyl (—C(CH3)2CH2CH3), 3-methyl-2-butyl (—CH(CH3)CH(CH3)2), 3-methyl-1-butyl (—CH2CH2CH(CH3)2), 2-methyl-1-butyl (—CH2CH(CH3)CH2CH3), 1-hexyl (—CH2CH2CH2CH2CH2CH3), 2-hexyl (—CH(CH3)CH2CH2CH2CH3), 3-hexyl (—CH(CH2CH3)(CH2CH2CH3)2), 2-methyl-2-pentyl (—C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (—CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl (—CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (—C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (—CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (—C(CH3)2CH(CH3)2), 3,3-dimethyl-2-butyl (—CH(CH3)C(CH3)3, 1-heptyl, 1-octyl, and the like.
The terms “alkyl” and the prefix “alk-” are inclusive of both straight chain and branched saturated carbon chain.
The term “alkylene” refers to a saturated divalent hydrocarbon group derived from a straight or branched chain saturated hydrocarbon by the removal of two hydrogen atoms, wherein the alkyl radical may be optionally substituted independently with one or more substituents described herein. Some non-limiting examples include methylene (—CH2—), ethylidene (—CH(CH3)—), ethylene (—CH2CH2—), isopropylene (—CH(CH3)CH2—), and the like.
The term “alkenyl” refers to linear or branched-chain monovalent hydrocarbon radical of two to twelve carbon atoms with at least one site of unsaturation, i.e., a carbon-carbon, sp2 double bond, wherein the alkenyl radical may be optionally substituted independently with one or more substituents described herein, and includes radicals having “cis” and “trans” orientations, or alternatively, “E” and “Z” orientations. Some non-limiting examples include ethylenyl or vinyl (—CH═CH2), allyl (—CH2CH═CH2), and the like.
The term “alkynyl” refers to linear or branched-chain monovalent hydrocarbon radical of two to twelve carbon atoms with at least one site of unsaturation, i.e., a carbon-carbon, sp triple bond, wherein the alkynyl radical may be unsubstituted or optionally substituted independently with one or more substituents described herein. Some non-limiting examples include ethynyl (—C≡CH), propynyl (propargyl, —CH2C≡CH), —C≡C—CH3, and the like.
The terms “haloalkyl”, “haloalkenyl” or “haloalkoxy” refers to alkyl, alkenyl, or alkoxy, as the case may be, substituted with one or more halogen atoms. Some non-limiting examples include —CF3, —OCF3, and the like.
The term “carbocycle”, “carbocyclyl”, “carbocyclic”, “carbocyclic ring” or “cycloaliphatic” refers to a monovalent or multivalent non-aromatic, saturated or partially unsaturated ring having 3 to 12 carbon atoms as a monocyclic ring system. Some non-limiting examples of suitable cycloaliphatic groups include cycloalkyl, cycloalkenyl, and cycloalkynyl. Further examples of cycloaliphatic groups include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexadienyl, and the like. The term “cycloalkyl” refers to a monovalent or multivalent saturated ring having 3 to 12 carbon atoms as a monocyclic ring system. The cycloalkyl groups herein are unsubstituted or optionally substituted independently with one or more substituents described herein.
The term “heterocycle”, “heterocyclyl”, “heterocyclic” or “heterocyclic ring” as used interchangeably herein refers to a monocyclic ring system in which one or more ring members are an independently selected heteroatom and that is completely saturated or that contains one or more units of unsaturation, but not aromatic having a single point of attachment to the rest of the molecule. One or more ring atoms are optionally substituted independently with one or more substituents described herein. In some embodiments, the “heterocycle”, “heterocyclyl”, “heterocyclic” or “heterocyclic ring” group is a monocycle having 3 to 7 ring members (2 to 6 carbon atoms and 1 to 3 heteroatoms selected from N, O, P, and S, wherein S or P is optionally substituted with one or more oxo to provide the group SO or SO2, PO or PO2 or a bicycle having 7 to 10 ring members (4 to 9 carbon atoms and 1 to 3 heteroatoms selected from N, O, P, and S, wherein the S or P is optionally substituted with one or more oxo to provide the group SO or SO2, PO or PO2.
The heterocyclyl may be a carbon radical or heteroatom radical. Some non-limiting examples of heterocyclic rings include pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, homo-piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinylimidazolinyl, imidazolidinyl, 1,2,3,4-tetrahydroiso-quinolinyl, 5-methyl-1,3-dioxol-4-yl. Examples of a heterocyclic group wherein 2 ring carbon atoms are substituted with oxo (═O) moieties are pyrimidindionyl and 1,1-dioxo-thiomorpholinyl. The heterocycle groups herein are unsubstituted or optionally substituted independently with one or more substituents described herein.
The term “heteroatom” refers to one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon, including any oxidized forms of nitrogen, sulfur, or phosphorus; the quaternized form of any basic nitrogen; or a substitutable nitrogen of a heterocyclic ring, for example N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR (as in N— substituted pyrrolidinyl).
The term “halogen” refers to F, Cl, Br, or I.
The term “H” refers to a single hydrogen atom. This radical may be attached, for example, to an oxygen atom to form a hydroxyl radical.
The term “D” or “2H” refers to a single deuterium atom. One of this radical may be attached, for example, to a methyl group to form a mono-deuterated methyl group (—CDH2), two of deuterium atoms may be attached to a methyl group to form a di-deuterated methyl (—CD2H), and three of deuterium atoms may be attached to a methyl group to form a tri-deuterated methyl group (—CD3).
The term “aryl” used alone or as part of a larger moiety as in “aryloxyalkyl” refers to monocyclic, bicyclic, and tricyclic carbocyclic ring systems having a total of six to fourteen ring members, wherein at least one ring in the system is aromatic, wherein each ring in the system contains 3 to 7 ring members and that has a single point of attachment to the rest of the molecule. The term “aryl” may be used interchangeably with the term “aryl ring”. Examples of aryl rings would include phenyl, naphthyl and anthracene.
The term “heteroaryl” used alone or as part of a larger moiety as in “heteroaralkyl” refers to monocyclic, bicyclic, and tricyclic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic, at least one ring in the system contains one or more heteroatoms, wherein each ring in the system contains 5 to 7 ring members and that has a single point of attachment to the rest of the molecule. The term “heteroaryl” may be used interchangeably with the term “heteroaryl ring” or the term “heteroaromatic”. The heteroaryl groups herein are unsubstituted or optionally substituted independently with one or more substituents described herein.
Some non-limiting examples of heteroaryl rings include the following monocycles: 2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, pyridazinyl (e.g., 3-pyridazinyl), 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (e.g., 5-tetrazolyl), triazolyl (e.g., 2-triazolyl and 5-triazolyl), 2-thienyl, 3-thienyl, pyrazolyl (e.g., 2-pyrazolyl), isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,3-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, pyrazinyl, 1,3,5-triazinyl, and the following bicycles: benzimidazolyl, benzofuryl, benzothiophenyl, indolyl (e.g., 2-indolyl), purinyl, quinolinyl (e.g., 2-quinolinyl, 3-quinolinyl, 4-quinolinyl), and isoquinolinyl (e.g., 1-isoquinolinyl, 3-isoquinolinyl or 4-isoquinolinyl).
The term “carboxy” or “carboxyl”, whether used alone or with other terms, such as “carboxyalkyl”, refers to —CO2H. The term “carbonyl”, whether used alone or with other terms, such as “aminocarbonyl”, refers to —(C═O)—. The term “acyl” refers to —(C═O)R, wherein R refers to alkyl.
The term “alkoxy” refers to an alkyl group, as previously defined, attached to the principal carbon chain through an oxygen (“alkoxy”) atom. Unless otherwise specified, alkoxy groups contain 1-20 carbon atoms. In some embodiments, alkoxy groups contain 1-8 carbon atoms. In other embodiments, alkoxy groups contain 1-6 carbon atoms. In still other embodiments, alkoxy groups contain 1-4 carbon atoms, and in yet other embodiments, alkoxy groups contain 1-3 carbon atoms.
Some non-limiting examples of alkoxy groups include methoxy (MeO, —OCH3), ethoxy (EtO, —OCH2CH3), 1-propoxy (n-PrO, n-propoxy, —OCH2CH2CH3), 2-propoxy (i-PrO, i-propoxy, —OCH(CH3)2), 1-butoxy (n-BuO, n-butoxy, —OCH2CH2CH2CH3), 2-methyl-1-propoxy (i-BuO, i-butoxy, —OCH2CH(CH3)2), 2-butoxy (s-BuO, s-butoxy, —OCH(CH3)CH2CH3), 2-methyl-2-propoxy (t-BuO, t-butoxy, —OC(CH3)3), 1-pentoxy(n-pentoxy, —OCH2CH2CH2CH2CH3), 2-pentoxy (—OCH(CH3)CH2CH2CH3), 3-pentoxy (—OCH(CH2CH3)2), 2-methyl-2-butoxy (—OC(CH3)2CH2CH3), 3-methyl-2-butoxy (—OCH(CH3)CH(CH3)2), 3-methyl-1-butoxy (—OCH2CH2CH(CH3)2), 2-methyl-1-butoxy (—OCH2CH(CH3)CH2CH3), and the like.
The term “alkylamino” embraces “N-alkylamino” and “N,N-dialkylamino” where amino groups are independently substituted with one alkyl radical and with two alkyl radicals, respectively. More preferred alkylamino radicals are “lower alkylamino” radicals having one or two alkyl radicals of one to six carbon atoms, attached to a nitrogen atom. Suitable alkylamino radicals may be mono or dialkylamino such as N-methylamino, N-ethylamino, N,N-dimethylamino, N,N-diethylamino, and the like.
The term “unsaturated” as used herein, refers to that a moiety has one or more units of unsaturation.
The term “comprising” is meant to be open ended, including the indicated component but not excluding other elements.
As described herein, a bond drawn from a substituent to the center of one ring within a ring system (as shown below) represents substitution of the substituent at any substitutable position on the rings to which it is attached. For example, Structure a represents possible substitution in any of the positions on the B ring shown in Structure b.




Unless otherwise stated, structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure; for example, the R and S configurations for each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention.
The term “tautomer” or “tautomeric form” refers to structural isomers of different energies which are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton, such as keto-enol and imine-enamine isomerizations. Valence tautomers include interconversions by reorganization of some of the bonding electrons.
Unless otherwise stated, all tautomeric forms of the compounds of the invention are within the scope of the invention. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms.
The term “prodrug” refers to a compound that is transformed in vivo into a compound of formula (I). Such a transformation can be affected, for example, by hydrolysis in blood or enzymatic transformation of the prodrug form to the parent form in blood or tissue. Prodrugs of the compounds of the invention may be, for example, esters. Esters that may be utilized as prodrugs in the present invention are phenyl esters, aliphatic C1-24 esters, acyloxymethyl esters, carbonates, carbamates, and amino acid esters. For example, a compound of the invention that contains an OH group may be acylated at this position in its prodrug form. Other prodrug forms include phosphates, such as, for example those phosphates resulting from the phosphonation of an OH group on the parent compound. A thorough discussion of prodrugs is provided in Higuchi et al., Pro-drugs as Novel Delivery Systems, Vol. 14, A.C.S. Symposium Series; Roche et al., ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987; Rautio et al., Prodrugs: Design and Clinical Applications, Nature Reviews Drug Discovery, 2008, 7, 255-270, and Hecker et al, Prodrugs of Phosphates and Phosphonates, J. Med. Chem., 2008, 51, 2328-2345, all of which are incorporated herein by reference.
A “metabolite” is a product produced through metabolism in the body of a specified compound or salt thereof. Metabolites of a compound may be identified using routine techniques known in the art and their activities determined using tests such as those described herein. Such products may result for example from the oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, and the like, of the administered compound.
Stereochemical definitions and conventions used herein generally follow Parker et al., ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York and Eliel et al., “Stereochemistry of Organic Compounds”, John Wiley & Sons, Inc., New York, 1994. The compounds of the invention may contain asymmetric or chiral centers, and therefore exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds of the invention, including but not limited to, diastereomers, enantiomers and atropisomers, as well as mixtures thereof such as racemic mixtures, form part of the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L, or R and S, are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes D and L or (+) and (−) are employed to designate the sign of rotation of plane-polarized light by the compound, with (−) or L meaning that the compound is levorotatory. A compound prefixed with (+) or D is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of one another. A specific stereoisomer may also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture or a racemate, which may occur where there has been no stereoselection or stereospecificity in a chemical reaction or process.
The term “racemic mixture” or “racemate” refers to an equimolar mixture of two enantiomeric species, devoid of optical activity.
A “pharmaceutically acceptable salt” refers to organic or inorganic salts of a compound of the invention. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmacol Sci, 1977, 66, 1-19, which is incorporated herein by reference. Some non-limiting examples of pharmaceutically acceptable, nontoxic salts include salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. The pharmaceutically acceptable salt is an alkali metal salt, an alkaline earth metal salt, an ammonium salt or a N+(C1-4 alkyl)4 salt. This invention also envisions the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. Water or oil-soluble or dispersable products may be obtained by such quaternization. Representative alkali or alkaline earth metal salts include sodium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, C1-8 sulfonate and aryl sulfonate.
A “solvate” refers to an association or complex of one or more solvent molecules and a compound of the invention. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine. The term “hydrate” refers to the complex where the solvent molecule is water.
The term “protecting group” or “PG” refers to a substituent that is commonly employed to block or protect a particular functionality while reacting other functional groups on the compound. For example, an “amino-protecting group” is a substituent attached to an amino group that blocks or protects the amino functionality in the compound. Suitable amino-protecting groups include acetyl, trifluoroacetyl, t-butoxy-carbonyl (BOC, Boc), benzyloxycarbonyl (CBZ, Cbz) and 9-fluorenylmethylenoxy-carbonyl (Fmoc). Similarly, a “hydroxy-protecting group” refers to a substituent of a hydroxy group that blocks or protects the hydroxy functionality. Suitable protecting groups include acetyl and silyl. A “carboxy-protecting group” refers to a substituent of the carboxy group that blocks or protects the carboxy functionality. Common carboxy-protecting groups include —CH2CH2SO2Ph, cyanoethyl, 2-(trimethylsilyl)ethyl, 2-(trimethylsilyl)ethoxymethyl, 2-(p-toluenesulfonyl)ethyl, 2-(p-nitrophenylsulfenyl)ethyl, 2-(diphenylphosphino)ethyl, nitroethyl and the like. For a general description of protecting groups and their use, see Greene et al., Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991 and Kocienski et al., Protecting Groups, Thieme, Stuttgart, 2005.
DESCRIPTION OF COMPOUNDS OF THE INVENTION
The present invention provides prodrugs of vemurafenib, salts, and pharmaceutical formulations thereof, which are potentially useful in the treatment of diseases, conditions and disorders modulated by protein kinases, especially B-Raf kinase. More specifically, the present invention provides a compound of Formula (I):




or a racemic mixture, a stereoisomer, a geometric isomer, a tautomer, a solvate, an N-oxide, a metabolite or a pharmaceutically acceptable salt thereof, wherein each of X and Y is as defined herein.

In some embodiments, each of X and Y is independently H, —C(═O)R3, —C(═O)OR4, —C(R1R2)OC(═O)R3, —C(R1R2)OC(═O)OR4 or —C(R1R2)OP(═O)(OR4)(OR4a), with the proviso that when Y is H, X is not —C(═O)CH3 or H;
each R1 and R2 is independently H, D, C1-6alkyl, C1-6haloalkyl, C3-6cycloalkyl, —(C1-4 alkylene)-(C3-6cycloalkyl), C3-6heterocyclyl or —(C1-4alkylene)-(C3-6heterocyclyl); or R1 and R2, together with the carbon atom they are attached to, form an unsubstituted or substituted 3-8 membered carbocyclic or heterocyclic ring;
each R3 is independently H, D, C1-10alkyl, C1-10haloalkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl or —(C1-6alkylene)-(5-10 membered heteroaryl), wherein each of the C1-10alkyl, C1-10haloalkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl and —(C1-6alkylene)-(5-10 membered heteroaryl) is unsubstituted or optionally substituted with 1, 2, 3 or 4 substituents independently selected from D, F, Cl, Br, I, C1-6alkyl, C1-6haloalkyl, —OH, —NH2, C1-6alkoxy and C1-6alkylamino; and
each R4 and R4a is independently H, C1-10alkyl, C1-10haloalkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl or —(C1-6alkylene)-(5-10 membered heteroaryl), wherein each of the C1-10alkyl, C1-10haloalkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl and —(C1-6alkylene)-(5-10 membered heteroaryl) is unsubstituted or optionally substituted with 1, 2, 3 or 4 substituents independently selected from D, F, Cl, OH, NH2, oxo (═O), C1-6alkyl, C1-6haloalkyl, C1-6alkoxy and C1-6alkylamino.
In other embodiments, each R1 and R2 is independently H, D or C1-3alkyl.
In other embodiments, each R3 is independently C1-10alkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl or —(C1-6alkylene)-(5-10 membered heteroaryl), wherein each of the C1-10alkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl and —(C1-6alkylene)-(5-10 membered heteroaryl) is unsubstituted or optionally substituted with 1, 2, 3 or 4 substituents independently selected from D, F, —OH, —OMe, —NH2, —NHMe, —NMe2 and C1-3alkyl.
In other embodiments, each R4 and R4a is independently H, C1-10alkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C1-3-8 heterocyclyl) or C6-10aryl, wherein each of the C1-10alkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl) and C6-10aryl is unsubstituted or optionally substituted with 1, 2, 3 or 4 substituents independently selected from D, F, OH, OMe, oxo (═O), NH2, —NHMe, —NMe2 and C1-3alkyl.
In other embodiments, each of X and Y is independently H or —C(R1R2)OP(═O)(OH)2, with the proviso that when Y is H, X is not H.
In other embodiments, each of X and Y is independently H, —C(═O)R3, —C(═O)OR4, —C(R1R2)OC(═O)R3 or —C(R1R2)OC(═O)OR4, with the proviso that when Y is H, X is not —C(═O)CH3 or H.
In other embodiments, the acyl moiety (—C(═O)R3) is derived from an α-amino acid or an optical isomer thereof by removing the OH group from the carboxy group of the α-amino acid.
In other embodiments, the α-amino acid is isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, alanine, asparagine, aspartate, glutamate, glutamine, proline, serine, para-tyrosine, arginine, histidine, cysteine, glycine, sarcosine, N,N-dimethyl glycine, homoserine, norvaline, norleucine, ornithine, homocysteine, homophenylalanine, phenylglycine, ortho-tyrosine, meta-tyrosine or hydroxyproline.
In other embodiments, the α-amino acid is isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, alanine, asparagine, aspartate, glutamate, glutamine, proline, serine, tyrosine, arginineor histidine, each of which has a S-configuration at the α-position.
In other embodiments, the pharmaceutically acceptable salt is an alkali metal salt, an alkaline earth metal salt, or an ammonium or N′(C1-4 alkyl)4 salt.
In other embodiments, the pharmaceutically acceptable salt is a sodium salt, a lithium salt, a potassium salt, a calcium salt, a magnesium salt, an ammonium salt, a quaternary ammonium salt, or a combination thereof.
In other embodiments, the pharmaceutically acceptable salt is a salt of an inorganic acid, an organic acid or a combination thereof, wherein the inorganic acid or the organic acid is hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, malic acid, 2-hydroxypropanic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, glucuronic acid, galacturonic acid, citric acid, tartaric acid, aspartic acid, glutamic acid, benzoic acid, cinnamic acid, p-toluenesulfonic acid, benzenesulfonic acid, mthanesulfonic acid, ethanesulfonic acid, trifluoromthanesulfonic acid or a combination thereof.
Some non-limiting examples of the compounds disclosed herein, and their pharmaceutically acceptable salts and solvates thereof, include:








TABLE 1











(1)


 






(2)


 






(3)


 






(4)


 






(5)


 






(6)


 






(7)


 






(8)


 






(9)


 






(10)


 






(11)


 






(12)


 






(13)


 






(14)


 






(15)


 






(16)


 






(17)


 






(18)


 






(19)


 






(20)


 






(21)


 






(22)


 






(23)


 






(24)


 






(25)


 






(26)


 






(27)


 






(28)


 






(29)


 






(30)


 






(31)


 






(32)


 






(33)


 






(34)


 






(35)


 






(36)


 






(37)


 






(38)


 






(39)


 






(40)


 






(41)


 






(42)









The present invention also comprises the use of a compound of the invention, or pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment either acutely or chronically of a hyperproliferative disease state and/or an angiogenesis mediated disease state, including those described previously. The compounds of the present invention are useful in the manufacture of an anti-cancer medicament. The compounds of the present invention are also useful in the manufacture of a medicament to attenuate or prevent disorders through inhibition of protein kinases. The present invention comprises a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I) in association with at least one pharmaceutically acceptable carrier, adjuvant or diluent.
The present invention also comprises a method of treating hyperproliferating and angiogenesis related disorders in a subject having or susceptible to such disorder, the method comprising treating the subject with a therapeutically effective amount of a compound of Formula (I).
Unless otherwise stated, all stereoisomers, geometric isomers, tautomers, solvates, metabolites, salts, and pharmaceutically acceptable prodrugs of the compounds of the invention are within the scope of the invention.
In certain embodiments, the salt is a pharmaceutically acceptable salt. The phrase “pharmaceutically acceptable” indicates that the substance or composition must be compatible chemically and/or toxicologically, with the other ingredients comprising a formulation, and/or the mammal being treated therewith.
The compounds of the invention also include salts of such compounds which are not necessarily pharmaceutically acceptable salts, and which may be useful as intermediates for preparing and/or purifying compounds of Formula (I) and/or for separating enantiomers of compounds of Formula (I).
Prodrug salts may be prepared in a variety of ways familiar to the skilled artisan, or according to the methods described herein. For example, a prodrug bearing an acidic prodrug group may be converted to the corresponding prodrug salt by treating with a base, followed by isolation through, for example, concentration, precipitation, or crystallization. If there exist more than one acidic hydrogen in the prodrug, then polyanionic salts may be prepared with the appropriate stoichiometric ration of base and prodrug. For example, a dianion may be formed from a prodrug containing a —P(═O)(OH)2 group with two equivalents of an appropriate base. Preferred salts of prodrugs of the invention include, but not limited to, sodium, potassium, calcium, magnesium, ammonium, and the like. Alternatively, an organic base may be used to prepare salts of the prodrug of the invention. Examples of preferred organic bases include, but are not limited to, lysine, arginine, N,N-diethylethanolamine, diethanolamine, ethanolamine, ethylenediamine, meglumine, morpholine, piperazine, piperidine, triethylamine, tromethamine (Tris), benzathine, benzene-ethanamine, adenine, cytosine and guanidine. As described in more detailed below, the prodrug salts may be formed in aqueous solution. Alternatively, they may be made in organic solution using and organic base such as sodium methoxide.
The cation may be exchanged by treating a solution of the desired prodrug anion with an appropriate cationic species to form an insoluble salt. The prodrug salt hydrates may be prepared, for example, by salt formation, isolation, or crystallization from aqueous solution.
In case that a prodrug bearing an basic group, the prodrug may be converted to the corresponding prodrug salt by treating the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, or with an organic acid, such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid, such as glucuronic acid or galacturonic acid, an alpha hydroxy acid, such as citric acid or tartaric acid, an amino acid, such as aspartic acid or glutamic acid, an aromatic acid, such as benzoic acid or cinnamic acid, a sulfonic acid, such as p-toluenesulfonic acid or ethanesulfonic acid, or the like.
Compounds disclosed herein are prodrugs. These compounds typically do not exhibit inhibitory activities against certain protein kinases in in vitro assays. However, the prodrug compounds are modified in vivo to produce a parent compound that has known inhibitory activities against certain protein kinases. Kinase inhibition activity of prodrug compounds is established by quantitating the blood or plasma levels of parent compound after prodrug administration. In vivo blood or plasma levels are reported as Cmax (maximum blood or plasma concentration) and AUC (area under curve from zero to a certain period of time or infinity, a measure of total drug exposure). Methods of measuring Cmax, AUC and other pharmacokinetic parameters in dogs and rats are provided in Example C.
Prodrug compounds are often, for example, better absorbed in vivo leading to higher overall blood or plasma levels of parent drug compound If both prodrug and parent compound are administered orally at the same doses.
Composition, Formulations and Administration of Compounds of The Invention
According to one aspect, the invention features pharmaceutical compositions that include a compound of formula (I), a compound listed in Table 1, and a pharmaceutically acceptable carrier, adjuvant, or vehicle. The amount of compound in the compositions of the invention is such that is effective to detectably inhibit a protein kinase in a biological sample or in a patient.
As described above, the pharmaceutically acceptable compositions of the present invention additionally comprise a pharmaceutically acceptable carrier, adjuvant, or vehicle, which, as used herein, includes any and all solvents, diluents, or other liquid vehicle, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like, as suited to the particular dosage form desired. Troy et al., Remington: The Science and Practice of Pharmacy, 21st, 2005, Lippincott Williams & Wilkins, Philadelphia and Swarbrick et al., Encyclopedia of Pharmaceutical Technology, eds. 1988-1999, Marcel Dekker, New York, all of which are herein incorporated by reference in their entireties, are disclosed various carriers used in formulating pharmaceutically acceptable compositions and known techniques for the preparation thereof. Except insofar as any conventional carrier medium is incompatible with the compounds of the invention such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutically acceptable composition, its use is contemplated to be within the scope of this invention.
Some non-limiting examples of materials which can serve as pharmaceutically acceptable carriers include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, or potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, wool fat, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols; such a propylene glycol or polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator.
The compositions of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. The term “parenteral” includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intraocular, intrahepatic, intralesional and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally or intravenously. Sterile injectable forms of the compositions of this invention may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium.
For this purpose, any bland fixed oil may be employed including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents that are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions. Other commonly used surfactants, such as TWEEN®, SPAN® and other emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.
Some non-limiting examples of the pharmaceutically acceptable compositions of this invention may be orally administered in any orally acceptable dosage form including capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.
Alternatively, the pharmaceutically acceptable compositions of this invention may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.
The pharmaceutically acceptable compositions of this invention may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.
Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches may also be used. For topical applications, the pharmaceutically acceptable compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Some non-limiting examples of carriers for topical administration of the compounds of this invention include mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutically acceptable compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Some non-limiting examples of suitable carriers include mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
For ophthalmic use, the pharmaceutically acceptable compositions may be formulated, e.g., as micronized suspensions in isotonic, pH adjusted sterile saline or other aqueous solution, or, preferably, as solutions in isotonic, pH adjusted sterile saline or other aqueous solution, either with or without a preservative such as benzylalkonium chloride. Alternatively, for ophthalmic uses, the pharmaceutically acceptable compositions may be formulated in an ointment such as petrolatum. The pharmaceutically acceptable compositions of this invention may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.
Some non-limiting examples of liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.
The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use. In order to prolong the effect of a compound of the present invention, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound then depends upon its rate of dissolution that, in turn, may depend upon crystal size and crystalline form. Alternatively, dissolving or suspending the compound in an oil vehicle accomplishes delayed absorption of a parenterally administered compound form.
Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide-polyglycolide. Depending upon the ratio of compound to polymer and the nature of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.
Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.
Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polythylene glycols and the like.
The active compounds can also be in micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. They may optionally contain pacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.
Dosage forms for topical or transdermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulation, eardrops, and eye drops are also contemplated as being within the scope of this invention. Additionally, the present invention contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.
The compounds of the invention are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The expression “dosage unit form” as used herein refers to a physically discrete unit of agent appropriate for the patient to be treated. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific effective dose level for any particular patient or organism will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed, and like factors well known in the medical arts.
The amount of the compounds of the present invention that may be combined with the carrier materials to produce a composition in a single dosage form will vary depending upon the host treated, the particular mode of administration. Preferably, the compositions should be formulated so that a dosage of between 0.01-200 mg/kg body weight/day of the inhibitor can be administered to a patient receiving these compositions.
Compounds of this invention can be administered as the sole pharmaceutical agent or in combination with one or more other additional therapeutic (pharmaceutical) agents where the combination causes no unacceptable adverse effects. This may be of particular relevance for the treatment of hyper-proliferative diseases such as cancer. In this instance, the compound of this invention can be combined with known cytotoxic agents, signal transduction inhibitors, or with other anti-cancer agents, as well as with admixtures and combinations thereof. As used herein, additional therapeutic agents that are normally administered to treat a particular disease, or condition, are known as “appropriate for the disease, or condition, being treated”. As used herein, “additional therapeutic agents” is meant to include chemotherapeutic agents and other anti-proliferative agents.
For example, chemotherapeutic agents or other antiproliferative agents may be combined with the compounds of this invention to treat proliferative disease or cancer. Some non-limiting examples of chemotherapeutic agents or other antiproliferative agents include HDAC inhibitors including SAHA, MS-275, MGO 103, and those described in WO 2006/010264, WO 03/024448, WO 2004/069823, US 2006/0058298, US 2005/0288282, WO 00/71703, WO 01/38322, WO 01/70675, WO 03/006652, WO 2004/035525, WO 2005/030705, WO 2005/092899, and demethylating agents including, but not limited to, 5-aza-dC, Vidaza and Decitabine and those described in U.S. Pat. No. 6,268,137, U.S. Pat. No. 5,578,716, U.S. Pat. No. 5,919,772, U.S. Pat. No. 6,054,439, U.S. Pat. No. 6,184,211, U.S. Pat. No. 6,020,318, U.S. Pat. No. 6,066,625, U.S. Pat. No. 6,506,735, U.S. Pat. No. 6,221,849, U.S. Pat. No. 6,953,783, U.S. Ser. No. 11/393,380.
In another embodiment of the present invention, for example, chemotherapeutic agents or other anti-proliferative agents may be combined with the compounds of this invention to treat proliferative diseases and cancer. Some non-limiting examples of known chemotherapeutic agents include other therapies or anticancer agents that may be used in combination with the inventive anticancer agents of the present invention and include surgery, radiotherapy (in but a few examples, gamma radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, and systemic radioactive isotopes, to name a few), endocrine therapy, taxanes (taxol, taxotere etc), platinum derivatives (cisplatin, carboplatin), biologic response modifiers (interferons, interleukins), tumor necrosis factor (TNF, TRAIL receptor targeting agents, to name a few), hyperthermia and cryotherapy, agents to attenuate any adverse effects (e.g., antiemetics), and other approved chemotherapeutic drugs, including, but not limited to, alkylating drugs (mechlorethamine, chlorambucil, cyclophosphamide, melphalan, ifosfamide), antimetabolites (methotrexate, pemetrexed etc), purine antagonists and pyrimidine antagonists (6-mercaptopurine, 5-fluorouracil, cytarabile, gemcitabine), spindle poisons (vinblastine, vincristine, vinorelbine), podophyllotoxins (etoposide, irinotecan, topotecan), antibiotics (doxorubicin, bleomycin, mitomycin), nitrosoureas (carmustine, lomustine), cell cycle inhibitors (KSP mitotic kinesin inhibitors, CENP-E and CDK inhibitors), enzymes (asparaginase), hormones (tamoxifen, leuprolide, flutamide, megestrol, dexamethasone), antiangiogenic agents (avastin and others), monoclonal antibodies (BENLYSTA®), brentuximab (ADCETRIS®), cetuximab (ERBITUX®), gemtuzumab (MYLOTARG®), ipilimumab (YERVOY®), ofatumumab (ARZERRA®), panitumumab (VECTIBIX®), ranibizumab (LUCERTIS®), rituximab (RITUXAN®), tositumomab (BEXXAR®), trastuzumab (HERCEPTIN®)), kinase inhibitors (imatinib (GLEEVEC®), sunitinib (SUTENT®), sorafenib (NEXAVAR®), erlotinib, (TARCEVA®), gefitinib (IRESSA®), dasatinib (SPRYCEL®), nilotinib (TASIGNA®), lapatinib (TYKERB®), crizotinib (XALKORI®), ruxolitinib (JAKAFI®), vemurafenib (ZELBORAF®), vandetanib (CAPRELSA®), pazopanib (VOTRIENT®), and others), and agents inhibiting or activating cancer pathways such as the mTOR, HIF (hypoxia induced factor) pathways (such as everolimus and temsirolimus) and others. For a more comprehensive discussion of updated cancer therapies see, http://www.nci.nih.gov/, a list of the FDA approved oncology drugs at http://www.fda.gov/cder/cancer/druglist-rame.htm, and The Merck Manual, Eighteenth Ed. 2006, all of which are herein incorporated by reference in their entireties.
In another embodiment, the compounds of the present invention can be combined, with cytotoxic anti-cancer agents. Examples of such agents can be found in the 13th Edition of the Merck Index (2001). These agents include, by no way of limitation, asparaginase, bleomycin, carboplatin, carmustine, chlorambucil, cisplatin, colaspase, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin, doxorubicin (adriamycine), epirubicin, etoposide, 5-fluorouracil, hexamethylmelamine, hydroxyurea, ifosfamide, irinotecan, leucovorin, lomustine, mechlorethamine, 6-mercaptopurine, mesna, methotrexate, mitomycin C, mitoxantrone, prednisolone, prednisone, procarbazine, raloxifen, streptozocin, tamoxifen, thioguanine, topotecan, vinblastine, vincristine and vindesine.
Other cytotoxic drugs suitable for use with the compounds of the invention include, but are not limited to, those compounds acknowledged to be used in the treatment of neoplastic diseases, such as those for example in Goodman and Gilman's The Pharmacological Basis of Therapeutics (Ninth Edition, 1996, McGraw-Hill). These agents include, by no way of limitation, aminoglutethimide, L-asparaginase, azathioprine, 5-azacytidine cladribine, busulfan, diethylstilbestrol, 2,2′-difluorodeoxycytidine, docetaxel, erythrohydroxynonyladenine, ethinyl estradiol, 5-fluorodeoxyuridine, 5-fluorodeoxyuridine monophosphate, fludarabine phosphate, fluoxymesterone, flutamide, hydroxyprogesterone caproate, idarubicin, interferon, medroxyprogesterone acetate, megestrol acetate, melphalan, mitotane, paclitaxel, pentostatin, N-phosphonoacetyl-L-aspartate (PALA), plicamycin, semustine, teniposide, testosterone propionate, thiotepa, trimethylmelamine, uridine, and vinorelbine.
Other cytotoxic anti-cancer agents suitable for use in combination with the compounds of the invention also include newly discovered cytotoxic principles such as oxaliplatin, vemurafenib, capecitabine, epothilone and its natural or synthetic derivatives, temozolomide (Quinn et al., J. Clin. Oncol., 2003, 21(4), 646-651), tositumomab (BEXXAR®), trabedectin (Vidal et al., Proceedings of the American Society for Clinical Oncology, 2004, 23, abstract 3181), and the inhibitors of the kinesin spindle protein Eg5 (Wood et al., Curr. Opin. Pharmacol., 2001, 1, 370-377).
In another embodiment, the compounds of the present invention can be combined with other signal transduction inhibitors. Examples of such agents include, by no way of limitation, antibody therapies such as trastuzumab (HERCEPTIN®), cetuximab (ERBITUX®), ipilimumab (YERVOY®) and pertuzumab. Examples of such therapies also include, by no way of limitation, small-molecule kinase inhibitors such as imatinib (GLEEVEC®), sunitinib (SUTENT®), sorafenib (NEXAVAR®), erlotinib (TARCEVA®), gefitinib (IRESSA®), dasatinib (SPRYCEL®), nilotinib (TASIGNA®), lapatinib (TYKERB®), crizotinib (XALKORI®), ruxolitinib (JAKAFI®), vemurafenib (ZELBORAF®), vandetanib (CAPRELSA®), pazopanib (VOTRIENT®), afatinib, alisertib, amuvatinib, axitinib, bosutinib, brivanib, canertinib, cabozantinib, cediranib, crenolanib, dabrafenib, dacomitinib, danusertib, dovitinib, foretinib, ganetespib, ibrutinib, iniparib, lenvatinib, linifanib, linsitinib, masitinib, momelotinib, motesanib, neratinib, niraparib, oprozomib, olaparib, pictilisib, ponatinib, quizartinib, regorafenib, rigosertib, rucaparib, saracatinib, saridegib, tandutinib, tasocitinib, telatinib, tivantinib, tivozanib, tofacitinib, trametinib, vatalanib, veliparib, vismodegib, volasertib, BMS-540215, BMS777607, JNJ38877605, TK1258, GDC-0941 (Folkes et al. J. Med. Chem. 2008, 51: 5522), BZE235, and others.
In another embodiment, the compounds of the present invention can be combined with inhibitors of histone deacetylase. Examples of such agents include, by no way of limitation, suberoylanilide hydroxamic acid (SAHA), LAQ-824 (Ottmann et al., Proceedings of the American Society for Clinical Oncology, 2004, 23, abstract 3024), LBH-589 (Beck et al., Proceedings of the American Society for Clinical Oncology, 2004, 23, abstract 3025), MS-275 (Ryan et al., Proceedings of the American Association of Cancer Research, 2004, 45, abstract 2452), FR-901228 (Piekarz et al., Proceedings of the American Society for Clinical Oncology, 2004, 23, abstract 3028) and MGCDOI 03 (U.S. Pat. No. 6,897,220).
In another embodiment, the compounds of the present invention can be combined with other anti-cancer agents such as proteasome inhibitors, and m-TOR inhibitors. These include, by no way of limitation, bortezomib, and CCI-779 (Wu et al., Proceedings of the American Association of Cancer Research 2004, 45, abstract 3849). The compounds of the present invention can be combined with other anti-cancer agents such as topoisomerase inhibitors, including but not limited to camptothecin.
Those additional agents may be administered separately from the compound-containing composition, as part of a multiple dosage regimen. Alternatively, those agents may be part of a single dosage form, mixed together with the compound of this invention in a single composition. If administered as part of a multiple dosage regimen, the two active agents may be submitted simultaneously, sequentially or within a period of time from one another which would result in the desired activity of the agents.
The amount of both the compound and the additional therapeutic agent (in those compositions which comprise an additional therapeutic agent as described above) that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. Normally, the amount of additional therapeutic agent present in the compositions of this invention will be no more than the amount that would normally be administered in a composition comprising that therapeutic agent as the only active agent. Preferably the amount of additional therapeutic agent in the presently disclosed compositions will range from about 50% to 100% of the amount normally present in a composition comprising that agent as the only therapeutically active agent. In those compositions which comprise an additional therapeutic agent, that additional therapeutic agent and the compound of this invention may act synergistically.
Uses of the Compounds and Effective Dosages
The invention features pharmaceutical compositions that include a compound of formula (I), or a compound listed in Table 1, and a pharmaceutically acceptable carrier, adjuvant, or vehicle. The amount of compound in the compositions of the invention is such that is effective to detectably inhibit a protein kinase, such as B-Raf kinase inhibitory activity. The compounds of the invention are useful in therapy as antineoplasia agents or to minimize deleterious effects of B-Raf signaling.
Compounds of the present invention would be useful for, but not limited to, the prevention or treatment of proliferative diseases, condition, or disorder in a patient by administering to the patient a compound or a composition of the invention in an effective amount. Such diseases, conditions, or disorders include cancer, metastatic cancer, atherosclerosis, and lung fibrosis.
Compounds of the invention would be useful for the treatment of neoplasia including cancer and metastasis, including, but not limited to: carcinoma such as cancer of the bladder, breast, colon, kidney, liver, lung (including small cell lung cancer), esophagus, gall-bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin (including squamous cell carcinoma); hematopoietic tumors of lymphoid lineage (including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell-lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkett's lymphoma); hematopoietic tumors of myeloid lineage (including acute and chronic myelogenous leukemias, myelodysplastic syndrome and promyelocytic leukemia); tumors of mesenchymal origin (including fibrosarcoma and rhabdomyosarcoma, and other sarcomas, e.g. soft tissue and bone); tumors of the central and peripheral nervous system (including astrocytoma, neuroblastoma, glioma and schwannomas); and other tumors (including melanoma, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma, thyroid follicular cancer and Kaposi's sarcoma).
Currently, vemurafenib is indicated for the treatment of patients with unresectable or metastatic melanoma with BRAFV600E mutation as detected by an US FDA-approved test.
Besides being useful for human treatment, these compounds are also useful for veterinary treatment of companion animals, exotic animals and farm animals, including mammals, rodents, and the like. More preferred animals include horses, dogs, and cats.
As used herein, the compounds of the present invention include the pharmaceutically acceptable derivatives thereof. Where the plural form is used for compounds, salts, and the like, this is taken to mean also a single compound, salt and the like.
The treatment method that includes administering a compound or composition of the invention can further include administering to the patient an additional therapeutic agent (combination therapy) selected from: a chemotherapeutic or anti-proliferative agent, or an anti-inflammatory agent, wherein the additional therapeutic agent is appropriate for the disease being treated and the additional therapeutic agent is administered together with a compound or composition of the invention as a single dosage form or separately from the compound or composition as part of a multiple dosage form. The additional therapeutic agent may be administered at the same time as a compound of the invention or at a different time. In the latter case, administration may be staggered by, for example, 6 hours, 12 hours, 1 day, 2 days, 3 days, 1 week, 2 weeks, 3 weeks, 1 month or 2 months.
The invention also features a method of inhibiting the growth of a cell that expresses mutated B-Raf, which includes contacting the cell with a compound or composition of the invention, thereby causing inhibition of growth of the cell. Examples of a cell whose growth can be inhibited include: a melanoma cell, a colorectal cancer cell, a breast cancer cell, a lung cancer cell, a papillary carcinoma cell, a prostate cancer cell, a lymphoma cell, a colon cancer cell, a pancreatic cancer cell, an ovarian cancer cell, a cervical cancer cell, a central nervous system cancer cell, an osteogenic sarcoma cell, a renal carcinoma cell, a hepatocellular carcinoma cell, a bladder cancer cell, a gastric carcinoma cell, a head and neck squamous carcinoma cell or a leukemia cell.
The invention provides a method of inhibiting mutated B-Raf kinase activity in a biological sample that includes contacting the biological sample with a compound or composition of the invention. The term “biological sample” means a sample outside a living organism and includes, without limitation, cell cultures or extracts thereof; biopsied material obtained from a mammal or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof. Inhibition of kinase activity, particularly mutated B-Raf kinase activity, in a biological sample is useful for a variety of purposes known to one of skill in the art. Non-limiting examples of such purposes include blood transfusion, organ-transplantation, biological specimen storage and biological assays.
In certain embodiments of the present invention an “effective amount” or “effective dose” of the compound or pharmaceutically acceptable composition is that amount effective for treating or lessening the severity of one or more of the aforementioned disorders. The compounds and compositions, according to the method of the present invention, may be administered using any amount and any route of administration effective for treating or lessening the severity of the disorder or disease. The exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the infection, the particular agent, its mode of administration, and the like. A compound or composition can also be administered with one or more other therapeutic agents, as discussed above.
Suitable dosage ranges for oral administration of the prodrug in the invention are dependent on the potency of vemurafenib in the particular indication of interest as well as the prodrug bioavailability. Dosage range may be readily determined by methods know to the artisan of ordinary skill. For example, vemurafenib is typically administered orally at a dose of 960 mg twice daily (bid) for the treatment of the indications of unresectable or metastatic melanoma with BRAFV600E mutation (as detected by an FDA-approved test).
The amount of a compound and/or pharmaceutical composition thereof that will be effective in the treatment of a particular disorder or condition disclosed herein can be determined by standard clinical techniques known in the art. In addition, in vitro or in vivo assays may optionally be employed to help identify optimal dosage ranges. The amount of a compound and/or pharmaceutical composition thereof administered will, of course, be dependent on, among other factors, the subject being treated (such as age, general health, sex, diet, weight, etc), the severity of the affliction, the manner, time and route of administration and the judgment of the prescribing physician.
The compounds are preferably assayed in vitro and in vivo, for the desired therapeutic or prophylactic activity, prior to use in humans. For example, in vitro assays can be used to determine whether administration of a specific compound or a combination of compounds is preferred. The compounds may also be demonstrated to be effective and safe using animal model systems. For example, vemurafenib is indicated for the treatment of patients with unresectable or metastatic melanoma with BRAFV600E mutation. Patients diagnosed with unresectable or metastatic melanoma were required to test their BARF mutation status before taken vemurafenib.
Preferably, a therapeutically effective dose of a compound and/or pharmaceutical composition thereof described herein will provide therapeutic benefit without causing substantial toxicity. Toxicity of compounds and/or pharmaceutical compositions thereof may be determined using standard pharmaceutical procedures and may be readily ascertained by the skilled artisan. The dose ratio between toxic and therapeutic effect is the therapeutic index. A compound and/or pharmaceutical composition thereof will preferably exhibit particularly high therapeutic indices in treating disease and disorders. The dosage of a compound and/or pharmaceutical composition thereof described herein will preferably be within a range of circulating concentrations that include an effective dose with minimal toxicity.
General Synthetic Procedures
In order to illustrate the invention, the following examples are included. However, it is to be understood that these examples do not limit the invention and are only meant to suggest a method of practicing the invention.
Generally, the compounds in this invention may be prepared by methods described herein, wherein the substituents are as defined for formula (I), above, except where further noted. The following non-limiting schemes and examples are presented to further exemplify the invention. Persons skilled in the art will recognize that the chemical reactions described herein may be readily adapted to prepare a number of other compounds of the invention, and alternative methods for preparing the compounds of this invention are deemed to be within the scope of this invention. For example, the synthesis of non-exemplified compounds according to the invention may be successfully performed by modifications apparent to those skilled in the art, e.g., by appropriately protecting interfering groups, by utilizing other suitable reagents known in the art other than those described, and/or by making routine modifications of reaction conditions. Alternatively, other reactions disclosed herein or known in the art will be recognized as having applicability for preparing other compounds of the invention.
In the examples described below, unless otherwise indicated all temperatures are set forth in degrees Celsius. Reagents were purchased from commercial suppliers such as Aldrich Chemical Company, Arco Chemical Company and Alfa Chemical Company, Shanghai Medpep. Co Ltd, Aladdin-Shanghai Jinchun Reagents, Ltd, and were used without further purification unless otherwise indicated. Common solvents were purchased from commercial suppliers such as Shantou XiLong Chemical Factory, Guangdong Guanghua Reagent Chemical Factory Co. Ltd., Guangzhou Reagent Chemical Factory, Tainjin YuYu Fine Chemical Ltd., Qingdao Tenglong Reagent Chemical Ltd., and Qingdao Ocean Chemical Factory.
Anhydrous THF, dioxane, toluene, and ether were obtained by refluxing the solvent with sodium Anhydrous CH2Cl2 and CHCl3 were obtained by refluxing the solvent with CaH2. EtOAc, PE, hexanes, DMA and DMF were treated with anhydrous Na2SO4 prior use.
The reactions set forth below were done generally under a positive pressure of nitrogen or argon or with a drying tube (unless otherwise stated) in anhydrous solvents, and the reaction flasks were typically fitted with rubber septa for the introduction of substrates and reagents via syringe. Glassware was oven dried and/or heat dried.
Column chromatography was conducted using a silica gel column. Silica gel (300-400 mesh) was purchased from Qingdao Ocean Chemical Factory. 1H NMR spectra were recorded with a Bruker 400 MHz spectrometer at ambient temperature. 1H NMR spectra were obtained as CDCl3, d6-DMSO, CD3OD or d6-acetone solutions (reported in ppm), using TMS (0 ppm) or chloroform (7.25 ppm) as the reference standard. When peak multiplicities are reported, the following abbreviations are used: s (singlet), d (doublet), t (triplet), m (multiplet), br (broadened), dd (doublet of doublets), dt (doublet of triplets). Coupling constants, when given, are reported in Hertz (Hz).
Low-resolution mass spectral (MS) data were generally determined on an Agilent 1200 or Agilent 6120 Series LCMS [Column: Zorbax SB-C18, 2.1×30 mm, 3.5 micorn, 6 minutes run, 0.6 mL/min flow rate. Mobile phase: 5-95% (0.1% formic acid in CH3CN) in (0.1% formic acid in H2O)] with UV detection at 210/254 nm and a low resonance electrospray mode (ESI).
Purities of compounds were assessed by Agilent 1100 Series high performance liquid chromatography (HPLC) with UV detection at 210 nm and 254 nm. Column was normally operated at 40° C.
The following abbreviations are used throughout the specification:

    BOC, Boc butyloxycarbonyl
    BSA bovine serum albumin
    BF3.Et2O boron Trifluoride-Ethyl Ether Complex
    CDCl3 chloroform deuterated
    CHCl3 chloroform
    CH2Cl2, DCM methylene chloride
    CH3SO2Cl, MsCl methanesulfonyl chloride
    Cs2CO3 cesium carbonate
    CuI copper(I) iodide
    DAST diethylaminosulfur trifluoride
    DBU 1,8-diazabicyclo[5.4.0]undec-7-ene
    DCE 1,2-dichloroethane
    DEAD dimethyl azodicarboxylate
    DIAD diisopropyl azodicarboxylate
    DIBAL diisobutylaluminum hydride
    DIEA, DIPEA diisopropylethylamine
    DMAP 4-dimethylaminopyridine
    DME 1,2-dimethoxyethane
    DMF dimethylformamide
    DMSO dimethylsulfoxide
    DPPA diphenylphosphoryl azide
    EDCI 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
    EtOAc, EA ethyl acetate
    Et2O diethyl ether
    Et3N, TEA triethylamine
    EtOCOCl ethyl chloroformate
    FBS fetal bovine serum
    G gram
    H hour
    Min minute
    HATU O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate
    HBr hydrobromic acid
    HBTU O-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium hexafluorophosphate
    HCl hydrochloric acid
    H2 hydrogen
    H2O water
    H2O2 hydrogen peroxide
    HOAc, AcOH acetic acid
    HOBt 1-hydroxybenzotriazole hydrate
    K2CO3 potassium carbonate
    KOH potassium hydroxide
    KH potassium hydride
    LiHMDS lithium bis(trimethylsilyl)amide
    NaHMDS sodium bis(trimethylsilyl)amide
    LDA lithium diisopropylamide
    MCPBA meta-chloroperbenzoic acid
    MeCN, CH3CN acetonitrile
    MeI methyl iodide
    MeOH, CH3OH methanol
    mL, ml milliliter
    N2 nitrogen
    NaBH3CN sodium cyanoborohydride
    NaCl sodium chloride
    NaH sodium hydride
    NaHCO3 sodium bicarbonate
    NaH2PO4 sodium biphosphate
    NaI sodium iodide
    NaO(t-Bu) sodium tert-butoxide
    NaOH sodium hydroxide
    Na2SO4 sodium sulfate
    NH3 ammonia
    NH4Cl ammonium chloride
    NMP N-methylpyrrolidinone
    PBS phosphate buffered saline
    P(t-Bu)3 tri(tert-butyl)phosphine
    Pd/C palladium on carbon
    Pd2(dba)3 bis(dibenzylideneacetone) palladium
    Pd(dppf)Cl2 1,1-bis(diphenylphosphino)ferrocene palladium chloride
    Pd(OAc)2 palladium acetate
    Pd(PPh3)4 palladium tetrakis triphenylphosphine
    PE petroleum ether (60-90° C.)
    POCl3 phosphorous oxychloride
    PyBop benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate
    RT, rt, r.t. room temperature
    Rt retention time
    5 nCl4 tin tetrachloride
    TBAB tetrabutylammonium bromide
    TBAHSO4 tetrabutylammonium hydrogen sulfate
    TBTU O-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium tetrafluoroborate TFA trifluoroacetic acid
    TEAC bis(tetra-ethylammonium)carbonate
    THF tetrahydrofuran
    μL microliters
    ZnCl2 zinc chloride


Outlined in Schemes 1 to 8 are representative general procedures for preparing the compounds disclosed herein. Each R1, R2, and M represents appropriate groups as defined herein, wherein each R1 is independently H or alkyl; each R2 is independently H, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl; M is alkali metal or alkaline earth metal; and n is 0, 1 or 2.




Some compounds having Formula (4a) or (4b) can be prepared by a general method illustrated in Scheme 1. Acid chloride (1) is reacted with an aldehyde (2) in the presence of a Lewis acid to furnish Chloroalkylester (3). Preferred Lewis acids include ZnCl2, 5 nCl4, BF3-Et2O, and others. The reaction is preferably run for between 30 min to 24 h neat or in an inert solvent such as DCM, CHCl3, C1CH2CH2Cl, and others, at a temperature ranging from −40° C. to 40° C. Chloroalkylester (33) is then coupled with vemurafenib under basic conditions to give the substituted azaindole product (4a), (4b) or (4a) and (4b). Preferred bases for the coupling reaction include KOH, NaOH, NaH, KH, sodium hexamethyldisilazide, potassium hexamethyldisilazide, lithium diisopropylamide (LDA), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), and others. The reaction is preferably run for a period in the range of 30 min to 24 h in an inert solvent such as DMF, DMSO, THF, dioxane, 1,2-dimethoxyethane (DME), and others, at a temperature ranging from −20° C. to 60° C.




Some compounds having Formula (8) can be prepared by a general method illustrated in Scheme 2. Chloroalkyl derived chloroformate (55) is reacted with alcohol (66) in the presence of a base to afford chloroalkyl carbonate (7). The preferred bases include pyridine, Et3N, i-Pr2NEt, 2,6-lutidine and others. The reaction is preferably performed in an inert solvent such as DCM, CHCl3, C1CH2CH2Cl and others, at a temperature ranging from −15° C. to 40° C. Condensation of carbonate (77) with vemurafenib with the aid of a base provide desired compound (88). Preferred bases for the coupling reaction include Et3N, KOH, NaOH, NaH, KH, sodium hexamethyldisilazide, potassium hexamethyldisilazide, lithium diisopropylamide (LDA), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) and others. The reaction is preferably run for a period in the range of 30 min to 24 h in an inert solvent such as DMF, DMSO, THF, dioxane, 1,2-dimethoxyethane (DME), and others, at a temperature ranging from −20° C. to 40° C.




Some compounds having Formula (12) can be prepared by a general method illustrated in Scheme 3. Chloroalkyl derived sulfochloridate (13) is reacted with protected amino acid (9) in the presence of a base and a catalyst such as TBAHSO4 to afford chloroalkyl ester (10). The preferred bases for the coupling reaction include NaHCO3, KHCO3, Na2CO3, K2CO3, Cs2CO3, Et3N, i-Pr2NEt, 2,6-lutidine, and others. The reaction is preferably performed in an inert solvent such as DCM, H2O, CHCl3, C1CH2CH2Cl, and others, at a temperature ranging from −20° C. to 40° C. Attachment of chloroalkyl ester (10) to vemurafenib is carried out under basic condition to provide protected aminoester (11). Preferred bases for the coupling reaction include KOH, NaOH, NaH or 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), and others. The reaction is preferably run for a period in the range of 30 min to 24 h in an inert solvent such as DMF, DMSO, THF, DCM, dioxane or 1,2-dimethoxyethane (DME) at a temperature ranging from −10° C. to 40° C. Amino-protecting groups include, but are not limited to, tert-butyloxycarbonyl (Boc), which may be removed under acidic conditions, for example by treatment with trifluoroacetic acid (TFA) in DCM, or HCl in ethyl acetate or ethyl ether.




Some compounds having Formula (17) or (1j) can be prepared by a general method illustrated in Scheme 4. Chloroalkyl derived sulfochloridate (13) is reacted with a suitably protected salt of a phosphoric acid diester derivative such as potassium di-tert-butyl phosphate (14) in the presence of a base and a catalyst such as TBAHSO4 to afford chloroalkyl di-tert-butyl phosphate (15). The preferred bases for the coupling reaction include NaHCO3, KHCO3, Na2CO3, K2CO3, Cs2CO3, Et3N, i-Pr2NEt, 2,6-lutidine, and others. The reaction is preferably performed in an inert solvent such as DCM, H2O, CHCl3, C1CH2CH2Cl, and others, at a temperature ranging from −20° C. to 40° C. Attachment of haloalkyl di-tert-butyl phosphate (15) to vemurafenib is carried out under basic condition to provide di-tert-butyl phosphate derivative (16). Preferred bases for the coupling reaction include KOH, NaOH, NaH or 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) and others. The reaction is preferably run for a period in the range of 30 min to 24 h in an inert solvent such as DMF, DMSO, THF, dioxane or 1,2-dimethoxyethane (DME) at a temperature ranging from −10° C. to 40° C. Tert-Butyl protecting group in compound (16) may be removed under acidic conditions, for example by treatment with trifluoroacetic acid (TFA) in DCM or HCl in ethyl acetate or ethyl ether. The phosphoric acid (17) is treated with a suitable base such as NaOH or KOH in aqueous solution to furnish phosphate salt (18).




Some compounds having Formula (19) can be prepared by a general method illustrated in Scheme 5. Acid chloride (1) is coupled with vemurafenib under basic conditions to give the desired product (19). Preferred bases for the coupling reaction include KOH, NaOH, Et3N, and others. The reaction is preferably run for a period in the range of 30 min to 24 h in an inert solvent such as DMF, DMSO, CHCl3, THF, dioxane, DCM, and others, at a temperature ranging from −20° C. to 40° C.




Some compounds having Formula (21) can be prepared by a general method illustrated in Scheme 6. Chloroformate (20) is coupled with vemurafenib under basic conditions to give the desired product (21). Preferred bases for the coupling reaction include KOH, NaOH, Et3N, and others. The reaction is preferably run for a period in the range of 30 min to 24 h in an inert solvent such as DMF, DMSO, CHCl3, THF, dioxane, DCM, and others, at a temperature ranging from −20° C. to 40° C.




Some compounds having Formula (24) can be prepared by a general method illustrated in Scheme 7. Under basic conditions, chloroformate (22) is condensed with vemurafenib in the presence of a suitable catalyst such as EDCI to give the desired Boc-protected azaindole product (23). Preferred bases for the condensation reaction include Et3N, 4-Phenylpyrimidine, and others. The reaction is preferably run for a period in the range of 30 min to 24 h in an inert solvent such as DMF, DME, dioxane, DCM, and others, at a temperature ranging from −10° C. to 40° C. Amino-protecting groups include, but are not limited to, tert-butyloxycarbonyl (Boc), which may be removed under acidic conditions, for example by treatment with trifluoroacetic acid (TFA) in DCM, or HCl in ethyl acetate or ethyl ether.




Some compounds having Formula (28) can be prepared by a general method illustrated in Scheme 8. Chloroalkyl derived sulfochloridate (13) is reacted with a amino-protected acid derivative (25) in the presence of a base and a catalyst such as TBAHSO4 to afford chloroalkyl ester (26). The preferred bases for the coupling reaction include NaHCO3, KHCO3, Na2CO3, K2CO3, Cs2CO3, Et3N, i-Pr2NEt, 2,6-lutidine and others. The reaction is preferably performed in an inert solvent such as DCM, H2O, CHCl3, C1CH2CH2Cl and others, at a temperature ranging from −20° C. to 40° C. Attachment of the haloalkyl ester (L) to vemurafenib is carried out under basic condition to provide substituted azaindole compound (27). Preferred bases for the coupling reaction include KOH, NaOH, NaH or 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) and others. The reaction is preferably run for a period in the range of 30 min to 24 h in an inert solvent such as DMF, DMSO, THF, dioxane or 1,2-dimethoxyethane (DME) at a temperature ranging from −10° C. to 40° C. Tert-butyl protecting group in substituted azaindole compound (27) may be removed under acidic conditions, for example by treatment with trifluoroacetic acid (TFA) in DCM or HCl in ethyl acetate or ethyl ether.
EXAMPLES
Example 1
ethyl 3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl(propylsulfonyl)carbamate




To a suspension of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)-propane-1-sulfonamide (0.2 g, 0.41 mmol) in dioxane (15 mL) were added ethyl chloroformate (EtOCOCl, 44 mg, 0.41 mmol) and triethylamine (0.14 g, 1.38 mmol). The reaction was stirred at rt for 1.5 h, then concentrated in vacuo. The residue was purified by a silica gel column chromatography (hexane/EtOAc (v/v)=4/1) to afford the title compound as a white solid (0.12 g, 50%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 562.1 [M+H]+;
1H NMR (400 MHz, CDCl3) δ 1.08 (t, J=7.2 Hz, 3H), 1.29 (t, J=7.2 Hz, 3H), 1.92-2.05 (m, 2H), 3.61 (br, 1H), 3.70 (br, 1H), 4.27-4.32 (q, J=7.2 Hz, 2H), 7.08-7.13 (m, 1H), 7.45-7.51 (m, 3H), 7.78 (d, J=8.4 Hz, 2H), 7.98 (d, J=2.0 Hz, 1H), 8.65 (d, J=1.9 Hz, 1H), 8.89 (d, J=2.0 Hz, 1H), 10.81 (s, 1H).
Example 2
N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)-N-(propylsulfonyl)benzamide




To a suspension of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)-propane-1-sulfonamide (0.2 g, 0.41 mmol) in dioxane (15 mL) were added benzoyl chloride (57 mg, 0.41 mmol) and triethylamine (0.14 g, 1.38 mmol). The reaction was stirred at rt for 1 h, then concentrated in vacuo. The residue was purified by a silica gel column chromatography (hexane/EtOAc (v/v)=4/1) to afford the title compound as a white solid (0.14 g, 55%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 594.1 [M+H]+;
1H NMR (400 MHz, CDCl3) δ 1.10 (t, J=7.4 Hz, 3H), 1.92-2.05 (m, 2H), 3.74 (t, J=7.6 Hz, 2H), 6.27-6.32 (q, J=7.2 Hz, 2H), 7.06-7.10 (m, 1H), 7.39 (t, J=7.6 Hz, 2H), 7.46-7.63 (m, 6H), 8.16 (d, J=7.0 Hz, 1H), 8.65 (d, J=2.1 Hz, 1H), 8.84 (d, J=2.0 Hz, 1H), 10.08 (s, 1H).
Example 3
N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)-N-(propylsulfonyl)isobutyramide




To a mixture of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.2 g, 0.41 mmol) and triethylamine (2 mL) in chloroform (10 mL) was added isobutyryl chloride (173 μA, 1.64 mmol) at rt. The reaction was stirred at rt for 16 h, then concentrated in vacuo. The residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=1/1 to 1/2) to afford the title compound as a white solid (2 mg, 0.8%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 560.1 [M+H]+;
1H NMR (400 MHz, CDCl3) δ 1.04-1.15 (t, J=7.4 Hz, 3H), 1.12 (s, 3H), 1.13 (s, 3H), 1.88-1.99 (m, 2H), 2.46-2.54 (m, 1H), 3.55-3.80 (m, 2H), 7.14-7.20 (m, 1H), 7.46-7.55 (m, 3H), 7.60-7.45 (d, J=8.4 Hz, 2H), 7.77 (s, 1H), 8.64-8.68 (d, J=2.0 Hz, 1H), 8.86-8.90 (m, 1H), 10.20 (s, 1H).
Example 4
N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)-N-(propylsulfonyl)acetamide




The title compound was prepared according to the procedure as described in Example 3 using N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.2 g, 0.41 mmol), triethylamine (1 mL) and acetyl chloride (116 μL, 1.64 mmol) in 1,4-dioxane (10 mL). The title compound was obtained as a white solid (69 mg, 31.8%). The title compound was characterized by LCMS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 532.1 [M+H]+;
1H NMR (400 MHz, CDCl3) δ 1.04-1.11 (t, J=7.4 Hz, 3H), 1.89-2.00 (m, 2H), 2.12 (s, 3H), 3.50-3.75 (m, 2H), 7.15-7.21 (m, 1H), 7.47-7.51 (m, 2H), 7.50-7.55 (m, 1H), 7.61-7.65 (m, 2H), 7.79 (s, 1H), 8.65-8.80 (d, J=2.1 Hz, 1H), 8.85-8.92 (m, 1H), 10.93 (s, 1H).
Example 5
N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)-N-(propylsulfonyl)nicotinamide




The title compound was prepared according to the procedure as described in Example 3 using N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.1 g, 0.2 mmol), triethylamine (228 μL, 1.6 mmol) and nicotinoyl chloride hydrochloride (146 mg, 0.2 mmol) in CHCl3 (10 mL). The crude product was purified by a silica gel column chromatography (PE/EtOAc (v/v)=2/1) to afford the title compound as a white solid (35 mg, 28.9%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 595.1 [M+H]+;
1H NMR (400 MHz, d6-DMSO) δ 1.02-1.08 (t, J=7.4 Hz, 3H), 1.83-1.93 (m, 2H), 3.86-3.93 (m, 2H), 7.37-7.44 (m, 1H), 7.44-7.50 (m, 1H), 7.55-7.60 (m, 2H), 7.74-7.82 (m, 2H), 7.96-8.04 (m, 1H), 8.25-8.29 (m, 1H), 8.59 (s, 1H), 8.64-8.67 (m, 1H), 8.72-8.75 (m, 1H), 8.77-8.80 (m, 1H), 9.06-9.09 (m, 1H), 13.11 (s, 1H).
Example 6
(S)-2-amino-N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)-3-methyl-N-(propylsulfonyl)butanamide hydrochloride




Step 1) (S)-tert-butyl (1-(N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propylsulfonamido)-3-methyl-1-oxobutan-2-yl)carbamate
To a mixture of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.3 g, 0.61 mmol), (S)-2-(Boc-amino)-3-methylbutyric acid (0.28 g, 1.29 mmol) and 4-ppy (9 mg, 0.06 mmol) in DMF (2 mL) was added EDCI (234 mg, 1.23 mmol). The reaction was stirred at rt for 16 h, then diluted with EtOAc (30 mL) and washed with water (30 mL×3). The mixture was dried over Na2SO4 and concentrated in vacuo. The residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=3/1 to 2/1) to afford the title compound as a white solid (41 mg, 29.1%). The title compound was characterized by LC-MS as shown below:

    LC-MS (ESI, neg. ion) m/z 686.6 [M−H]−.


Step 2) (S)-2-amino-N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)-3-methyl-N-(propylsulfonyl)butanamide hydrochloride
A mixture of (S)-tert-butyl (1-(N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propylsulfonamido)-3-methyl-1-oxobutan-2-yl)carbamate (50 mg, 0.07 mmol) and a saturated solution of HCl in EtOAc (2 mL) was stirred at rt for 2 h. The mixture was concentrated in vacuo to afford the title compound as a white solid (49 mg, 99%). The title compound was characterized by LC-MS as shown below:
LC-MS (ESI, pos. ion) m/z 589.2 [M+H]+.
Example 7
7a (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl pivalate
7b (N-(3-(5-(4-chlorophenyl)-1-((pivaloyloxy)methyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propylsulfonamido)methyl pivalate




To a mixture of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.1 g, 0.2 mmol) and Et3N (57 μL, 0.41 mmol) in DMF (1 mL) was added chloromethyl pivalate (35 μL, 0.25 mmol) at rt. The reaction was stirred at rt for 24 h, then concentrated in vacuo. The residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=4/1 to 1/1) to afford (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl pivalate (30 mg, 23%) and (N-(3-(5-(4-chlorophenyl)-1-((pivaloyloxy)methyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl) propylsulfonamido) methyl pivalate (20 mg, 13.3%) as white solids. The title compounds were characterized by LC-MS and 1H NMR as shown below:
7a (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl pivalate
LC-MS (ESI, pos. ion) m/z 604.1 [M+H]+;
1H NMR (400 MHz, CDCl3) δ 1.04-1.10 (t, J=7.4 Hz, 3H), 1.16 (s, 9H), 1.86-1.96 (m, 2H), 3.10-3.16 (m, 2H), 6.27 (s, 2H), 6.58-6.65 (br, 1H), 7.04-7.10 (m, 1H), 7.46-7.51 (m, 2H), 7.59-7.63 (m, 2H), 7.69-7.77 (m, 1H), 7.84 (s, 1H), 8.67-8.70 (d, J=2.2 Hz, 1H), 8.84-8.87 (d, J=2.2 Hz, 1H).
7b (N-(3-(5-(4-chlorophenyl)-1-((pivaloyloxy)methyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propylsulfonamido)methyl pivalate
LC-MS (ESI, pos. ion) m/z 718.2 [M+H]+;
1H NMR (400 MHz, CDCl3) δ 1.02-1.08 (t, J=7.4 Hz, 3H), 1.15 (s, 9H), 1.18 (s, 9H), 1.87-1.97 (m, 2H), 3.13-3.19 (m, 2H), 5.65 (s, 2H), 6.28 (s, 2H), 7.06-7.12 (m, 1H), 7.45-7.50 (m, 2H), 7.54-7.60 (m, 1H), 7.59-7.64 (m, 2H), 7.92 (s, 1H), 8.66-8.69 (d, J=2.2 Hz, 1H), 8.84-8.87 (d, J=2.1 Hz, 1H).
Example 8
(N-(3-(1-(acetoxymethyl)-5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propylsulfonamido)methyl acetate




Step 1) chloromethyl acetate
To a mixture of acteyl chloride (10 mL, 94.6 mmol) and ZnCl2 (25 mg, 186 mmol) was added paraformaldehyde (2.84 g, 94.6 mmol). The reaction was stirred at rt for 40 min. It was then heated to 60° C. and stirred further for 16 h. The mixture was cooled to rt and filtered through a short silica gel column (100% PE). The solution was concentrate in vacuo at 30° C. and the resulting oil was distilled at 60° C. under reduced pressure to give the title compound as colorless oil (2.34 g, 15.3%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.18-1.23 (d, J=7.0 Hz, 6H), 2.56-2.68 (m, 1H), 5.72 (s, 2H).
Step 2) (N-(3-(1-(acetoxymethyl)-5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propylsulfonamido)methyl acetate
To a mixture of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.1 g, 0.2 mmol) and Et3N (57 μL, 0.4 mmol) in DMF (1 mL) was added chloromethyl acetate (23 μL, 0.3 mmol) at rt. The reaction was stirred at rt for 16 h, and then concentrated in vacuo. The residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=2/1 to 1/1) to afford the title compound as a white solid (52 mg, 40.3%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 634.1 [M+H]+;
1H NMR (400 MHz, CDCl3) δ 1.03-1.09 (t, J=7.4 Hz, 3H), 2.07-2.12 (d, J=4.9 Hz, 2H), 3.13-3.17 (m, 2H), 5.64 (s, 2H), 6.28 (s, 2H), 7.07-7.13 (m, 1H), 7.46-7.51 (m, 2H), 7.56-7.64 (m, 3H), 7.93 (s, 1H), 8.67-8.70 (d, J=2.2 Hz, 1H), 8.85-8.88 (d, J=2.2 Hz, 1H).
Example 9
(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl isobutyrate




Step 1) chloromethyl isobutyrate
The title compound was prepared according to the procedure as described in Example 8 Step 1 using isobutyryl chloride (10 mL, 94.6 mmol), ZnCl2 (25 mg, 186 mmol) and paraformaldehyde (2.84 g, 94.6 mmol). The title compound was obtained as colorless oil (5.52 g, 42.9%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.18-1.23 (d, J=7.0 Hz, 6H), 2.56-2.68 (m, 1H), 5.72 (s, 2H).
Step 2) (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl isobutyrate
To a mixture of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.1 g, 0.2 mmol) and Et3N (57 μL, 0.4 mmol) in DMF (1 mL) was added chloromethyl isobutyrate (34 mg, 0.25 mmol) at rt. The reaction was stirred at rt for 16 h, then concentrated in vacuo. The residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=4/1 to 3/1) to afford the title compound as a white solid (45 mg, 37.5%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 590.1 [M+H]+;
1H NMR (400 MHz, CDCl3) δ 1.04-1.10 (t, J=7.4 Hz, 3H), 1.85-1.96 (m, 2H), 2.54-2.62 (m, 1H), 3.09-3.15 (m, 2H), 6.27 (s, 2H), 6.51 (s, 1H), 7.04-7.10 (m, 1H), 7.46-7.51 (m, 2H), 7.58-7.63 (m, 2H), 7.70-7.77 (m, 1H), 7.84 (s, 1H), 8.67-8.70 (d, J=2.2 Hz, 1H), 8.85-8.88 (d, J=2.2 Hz, 1H).
Example 10
1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl propionate




Step 1) 1-chloroethyl propionate
To a mixture of propionylchloride (10 mL, 114.56 mmol) and zinc (II) chloride (25 mg, 186 mmol) was added acetaldehyde (7.76 mL, 137.48 mmol) at 0° C. The reaction was stirred at rt for 16 h, and then filtered through a short silica gel column (100% PE). The solution was concentrated in vacuo at 20° C. and the resulting oil was distilled at 25-27° C. under reduced pressure to give the title compound as colorless oil (4.7 g, 30%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.14-1.20 (t, J=7.6 Hz, 3H), 1.77-1.80 (d, J=5.8 Hz, 3H), 2.41 (q, J=7.6 Hz, 2H), 6.51-6.60 (q, J=5.8 Hz, 1H).
Step 2) 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl propionate
To a solution of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.2 g, 0.41 mmol) in DMF (1 mL) was added Et3N (165 mg, 1.64 mmol). The reaction was stirred at 10° C. for 10 min, followed by the dropwise addition of a solution of TBAB (264 mg, 1.64 mmol) in DMF (0.5 mL) and a solution of 1-chloroethyl propionate (67 mg, 0.41 mmol) in DMF (0.5 mL) over 30 min. The mixture was stirred at 10° C. for another 3 h, then diluted with EtOAc (40 mL) and filtered. The filtrate was concentrated in vacuo and the residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=4/1) to afford the title compound as a white solid (141 mg, 58%). The title compound was characterized by LCMS and 1H NMR as shown below:
LC-MS (ESI, neg. ion) m/z 588.3 [M−H]−;
1H NMR (400 MHz, CDCl3) δ 1.00-1.17 (m, 6H), 1.85-2.00 (m, 5H), 2.26-2.41 (m, 2H), 3.08-3.17 (m, 2H), 6.50-6.80 (br, 1H), 7.03-7.12 (m, 1H), 7.35-7.42 (m, 1H), 7.42-7.51 (d, J=8.2 Hz, 2H), 7.56-7.64 (d, J=8.4 Hz, 2H), 7.65-7.76 (m, 1H), 7.77 (s, 1H), 8.63-8.70 (m, 1H), 8.77-8.83 (m, 1H).
Example 11
1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)-2-methylpropyl propionate




Step 1) 1-chloro-2-methylpropyl propionate
To a mixture of propionyl chloride (10 mL, 115 mmol) and zinc (II) chloride (25 mg, 186 mmol) was added isobutyraldehyde (12.5 mL, 138 mmol) at −20° C. The reaction was stirred at 0° C. for 2 h, then at rt for another 16 h. The resulting mixture was filtered through a short silica gel column (100% PE). The solution was concentrated in vacuo at 40° C. and the resulted oil was distilled at 90° C. under reduced pressure to give the title compound as colorless oil (0.52 g, 2.7%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.03-1.08 (m, 6H), 1.15-1.20 (t, J=7.5 Hz, 3H), 2.10-2.20 (m, 1H), 2.35-2.45 (q, J=7.6 Hz, 1H), 6.30-6.32 (d, J=4.7 Hz, 1H).
Step 2) 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)-2-methylpropyl propionate
The title compound was prepared according to the procedure as described in Example 10 Step 2 using a solution of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.2 g, 0.4 mmol) in DMF (1 mL), KOH (46 mg, 0.80 mmol), a solution of TBAB (264 mg, 0.81 mmol) in DMF (0.5 mL) and a solution of 1-chloro-2-methylpropyl propionate (81 mg, 0.49 mmol) in DMF (0.5 mL). The title compound was obtained as a white solid (114 mg, 45%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, neg. ion) m/z 616.3 [M−H]−;
1H NMR (400 MHz, CDCl3) δ 0.77-0.85 (d, J=6.8 Hz, 3H), 1.03-1.20 (m, 9H), 2.30-2.48 (m, 2H), 2.68-2.80 (m, 1H), 3.09-3.17 (m, 2H), 4.12-4.27 (m, 2H), 6.66 (br, 1H), 6.87-6.83 (d, J=9.2 Hz, 1H), 7.03-7.10 (m, 1H), 7.44-7.50 (d, J=8.4 Hz, 2H), 7.57-7.63 (d, J=8.4 Hz, 2H), 7.68-7.77 (m, 2H), 8.65-8.70 (d, J=2.0 Hz, 1H), 8.78-8.83 (m, 1H).
Example 12
1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl pentanoate




Step 1) 1-chloroethyl pentanoate
To a mixture of pentanoyl chloride (5 mL, 41.5 mmol) and zinc (II) chloride (25 mg, 186 mmol) was added acetaldehyde (2 mL, 49.5 mmol) at −20° C. The reaction was stirred at 0° C. for 1 h, then stirred further at rt for 2 h. The mxiture was filtered through a short silica gel column (100% PE). The solution was concentrated in vacuo at 40° C. and the resulting oil was distilled at 60° C. under reduced pressure to give the title compound as colorless oil (5.5 g, 81%). The title compound was characterized by
1H NMR and 13C NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 0.90-0.94 (t, J=7.3 Hz, 3H), 1.32-1.40 (m, 2H), 1.58-1.65 (m, 2H), 1.77-1.79 (d, J=5.8 Hz, 3H), 2.33-2.37 (t, J=7.5 Hz, 2H), 6.52-6.57 (q, J=5.8 Hz, 1H);
13C NMR (100 MHz, CDCl3) δ 13.7, 22.2, 25.2, 26.7, 33.9, 80.6, 171.5.
Step 2) 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl pentanoate
To a suspension of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.3 g, 0.61 mmol) in anhydrous DMF (2.5 mL) was added KOH (69 mg, 1.23 mmol). The reaction was stirred at 10° C. for 30 min, followed by adding a solution of 1-chloroethyl pentanoate (0.1 g, 0.61 mmol) in anhydrous DMF (0.5 mL) dropwise over 30 min. The mixture was stirred at 10° C. for another 16 h, then diluted with EtOAc (40 mL) and filtered. The filtrate was concentrated in vacuo and the residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=7/1) to give the crude product. The crude product was recrystallized in PE/EtOAc (10/1) to give the title compound as a white solid (0.14 g, 37.8%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, neg. ion) m/z 615.5 [M−H]−;
1H NMR (400 MHz, CDCl3) δ 0.83-0.87 (t, J=7.3 Hz, 3H), 1.04-1.08 (t, J=7.4 Hz, 3H), 1.24-1.30 (m, 2H), 1.53-1.59 (m, 2H), 1.86-1.92 (m, 2H), 1.92-1.94 (d, J=6.3 Hz, 3H), 2.29-2.34 (m, 2H), 3.10-3.14 (m, 2H), 6.53 (br, 1H), 7.05-7.10 (m, 1H), 7.35-7.40 (q, J=6.3 Hz, 1H), 7.46-7.48 (d, J=8.5 Hz, 2H), 7.58-7.60 (d, J=8.5 Hz, 2H), 7.68-7.74 (td, J=8.9, 5.5 Hz, 1H), 7.76 (s, 1H), 8.66-8.67 (d, J=2.1 Hz, 1H), 8.80-8.81 (d, J=2.0 Hz, 1H).
Example 13
1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl cyclopentanecarboxylate




Step 1) 1-chloroethyl cyclopentanecarboxylate
To a mixture of cyclopentanecarbonyl chloride (5 g, 37.7 mmol) and zinc (II) chloride (25 mg, 186 mmol) was added acetaldehyde (1.83 g, 41.5 mmol) at −20° C. The reaction was stirred at 0° C. for 1 h, then at r.t for another 2 h. The resulting mixture was filtered through a short silica gel column (100% DCM). The solution was concentrated in vacuo at 40° C. and the resulted oil was distilled at 55° C. under reduced pressure to give the title compound as colorless oil (5.6 g, 85%). The title compound was characterized by 1H NMR and 13C NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.57-1.62 (m, 2H), 1.68-1.72 (m, 2H), 1.78-1.79 (d, J=5.8 Hz, 3H), 1.80-1.93 (m, 4H), 2.69-2.80 (m, 1H), 6.52-6.57 (q, J=5.8 Hz, 1H);
13C NMR (100 MHz, CDCl3) δ 25.2, 25.9, 29.9, 30.0, 43.8, 80.8, 183.5.
Step 2) 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl cyclopentanecarboxylate
The title compound was prepared according to the procedure as described in Example 12 Step 2 using N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.3 g, 0.61 mmol) in anhydrous DMF (2.5 mL), KOH (69 mg, 1.23 mmol) and a solution of 1-chloroethyl cyclopentanecarboxylate (108 mg, 0.61 mmol) in anhydrous DMF (0.5 mL). The curde product was purified by a silica gel column chromatography (PE/EtOAc (v/v)=5/1) and recrystallized in PE to give the title compound as a white solid (147 mg, 38%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, neg. ion) m/z 627.5 [M−H]−;
1H NMR (400 MHz, CDCl3) δ 1.04-1.08 (t, J=7.4 Hz, 3H), 1.23-1.29 (m, 2H), 1.29-1.33 (m, 2H), 1.61-1.66 (m, 2H), 1.77-1.81 (m, 2H), 1.86-1.89 (m, 2H), 1.90-1.92 (d, J=6.2 Hz, 3H), 2.71-2.75 (m, 1H), 3.10-3.14 (m, 2H), 6.47 (br, 1H), 7.05-7.10 (m, 1H), 7.35-7.40 (q, J=6.3 Hz, 1H), 7.46-7.48 (d, J=8.5 Hz, 2H), 7.58-7.60 (d, J=8.4 Hz, 2H), 7.70-7.74 (td, J=9.0, 5.5 Hz, 1H), 7.75 (s, 1H), 8.66-8.67 (d, J=2.1 Hz, 1H), 8.80-8.81 (d, J=2.1 Hz, 1H).
Example 14
1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl isobutyrate




Step 1) 1-chloroethyl isobutyrate
To a mixture of isobutyryl chloride (10 mL, 94.7 mmol) and zinc (II) chloride (25 mg, 186 mmol) was added acetaldehyde (5 g, 113.6 mmol) at −20° C. The reaction was stirred at 0° C. for 16 h, then at rt for another 2 h. The mixture was filtered through a short silica gel column (100% PE). The solution was concentrated in vacuoat 30° C. and the resulting oil was distilled at 65° C. under reduced pressure to give the title compound as colorless oil (2.64 g, 18.5%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.15-1.25 (m, 6H), 1.77-1.82 (d, J=5.8 Hz, 3H), 2.50-2.63 (m, 1H), 6.50-6.60 (q, J=5.8 Hz, 1H).
Step 2) 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl isobutyrate
To a mixture of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.5 g, 1.02 mmol) and potassium hydroxide (115 mg, 2.04 mmol) in anhydrous DMF (4 mL) was added a solution of chloromethyl isobutyrate (37 mg, 0.25 mmol) in anhydrous DMF (1 mL) over 30 min at 10° C. The reaction was stirred at 10° C. for 3 h, then diluted with EtOAc (40 mL) and filtered. The filtrate was concentrated in vacuo and the residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=5/1 to 4/1) to afford the title compound as a white solid (0.39 g, 63.3%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 626.0 [M+Na]+;
1H NMR (400 MHz, CDCl3) δ 1.03-1.12 (m, 6H), 1.13-1.20 (d, J=7.0 Hz, 3H), 1.85-1.97 (m, 5H), 2.50-2.60 (m, 1H), 3.09-3.17 (m, 2H), 6.55 (s, 1H), 7.04-7.10 (m, 1H), 7.35-7.42 (m, 1H), 7.46-7.51 (d, J=8.4 Hz, 2H), 7.57-7.63 (d, J=8.4 Hz, 2H), 7.68-7.75 (m, 1H), 7.76 (s, 1H), 8.67-8.70 (d, J=2.1 Hz, 1H), 8.78-8.84 (d, J=1.8 Hz, 1H).
Example 15
(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl propionate




Step 1) chloromethyl propionate
To a mixture of paraformaldehyde (3.37 g, 112.3 mmol) and zinc (II) chloride (25 mg, 186 mmol) was added propionyl chloride (10 mL, 112.3 mmol). The reaction was stirred at rt for 40 min, then heated to 60° C. and stirred further for 16 h. The mixture was cooled to rt and filtered through a short silica gel column (100% PE). The solvent was removed in vacuo at 20° C. and the resulting oil was distilled at 40° C. under reduced pressure to give the title compound as colorless oil (9.8 g, 70%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.16-1.19 (d, J=7.56 Hz, 3H), 2.39-2.45 (t, J=7.56 Hz, 2H), 5.71 (s, 2H).
Step 2) (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl propionate
The title compound was prepared according to the procedure as described in Example 14 Step 2 using N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.3 g, 0.61 mmol) in anhydrous DMF (2.5 mL), KOH (69 mg, 1.23 mmol) and a solution of chloromethyl propionate (75 mg, 0.61 mmol) in anhydrous DMF (0.5 mL). The crude product was purified by a silica gel column chromatography (PE/EtOAc (v/v)=4/1), and recrystallized in methanol to afford the title compound as a white solid (145 mg, 41.4%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, neg. ion) m/z 574.1 [M−H]−;
1H NMR (400 MHz, CDCl3) δ 1.04-1.08 (t, J=7.4 Hz, 3H), 1.09-1.13 (t, J=7.5 Hz, 3H), 1.87-1.93 (m, 2H), 2.34-2.40 (d, J=7.5 Hz, 2H), 3.09-3.13 (m, 2H), 6.27 (s, 2H), 6.63 (br, 1H), 7.04-7.09 (m, 1H), 7.45-7.49 (m, 2H), 7.58-7.61 (m, 2H), 7.70-7.75 (td, J=5.5, 9.0 Hz, 1H), 7.85 (s, 1H), 8.67-8.68 (d, J=2.2 Hz, 1H), 8.84-8.85 (d, J=2.1 Hz, 1H).
Example 16
1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)-2-methylpropyl isobutyrate




Step 1) 1-chloro-2-methylpropyl isobutyrate
To a mixture of isobutyryl chloride (10 mL, 94.7 mmol) and zinc (II) chloride (25 mg, 186 mmol) was added isobutyraldehyde (10.4 mL, 113.6 mmol) at −20° C. The reaction was stirred at 0° C. for 1 h, then at rt for another 2 h. The mixture was filtered through a short silica gel column (100% PE). The solution was concentrated in vacuo at 40° C. and the resulted oil was distilled at 75° C. under reduced pressure to give the title compound as colorless oil (13.48 g, 79.3%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.04-1.08 (dd, J=3.68, 3.08 Hz, 6H), 1.19-1.22 (d, J=7.04 Hz, 6H), 2.12-2.21 (m, 1H), 2.57-2.65 (m, J=7.04 Hz, 1H), 6.29-6.31 (d, J=4.72 Hz, 1H).
Step 2) 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)-2-methylpropyl isobutyrate
The title compound was prepared according to the procedure as described in Example 14 Step 2 using N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.3 g, 0.61 mmol) in anhydrous DMF (2.5 mL), KOH (69 mg, 1.23 mmol) and a solution of 1-chloro-2-methylpropyl isobutyrate (108 mg, 0.61 mmol) in anhydrous DMF (0.5 mL). The crude product was purified by a silica gel column chromatography (PE/EtOAc (v/v)=4/1), and recrystallized in methanol to afford the title compound as a white solid (47 mg, 13.4%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, neg. ion) m/z 629.5 [M−H]−;
1H NMR (400 MHz, CDCl3) δ 1.04-1.08 (t, J=7.4 Hz, 3H), 1.08-1.12 (m, 6H), 1.86-1.95 (m, 2H), 2.54-2.61 (m, 1H), 3.09-3.14 (m, 2H), 6.27 (s, 2H), 6.67 (br, 1H), 7.04-7.09 (m, 1H), 7.46-7.49 (m, 2H), 7.59-7.61 (m, 2H), 7.70-7.75 (td, J=5.5, 9.0 Hz, 1H), 7.85 (s, 1H), 8.67-8.68 (d, J=2.1 Hz, 1H), 8.84-8.85 (d, J=2.1 Hz, 1H).
Example 17
(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-(piperidin-4-yl)acetate hydrochloride




Step 1) tert-butyl 4-(2-(chloromethoxy)-2-oxoethyl)piperidine-1-carboxylate
To a solution of 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)acetic acid (1 g, 4.1 mmol) in DCM (25 mL) and H2O (25 mL) were added NaHCO3 (1.38 g, 16.4 mmol) and TBAHSO4 (0.14 g, 0.41 mmol). The reaction was stirred at 0° C. for 10 min, followed by the addition of a solution of chloromethyl sulfochloridate (499 μL, 5.5 mmol) in DCM (5 mL). The mixture was stirred at rt for 20 h, then washed with brine (25 mL). The organic phase was dried over anhydrous Na2SO4 and concentrated in vacuo. The residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=10/1) to give the title compound as colorless oil (826 mg, 69.1%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.11-1.24 (m, 2H), 1.65-1.74 (m, 2H), 1.90-2.03 (m, 1H), 2.30-2.35 (d, J=7.1 Hz, 2H), 2.66-2.70 (m, 2H), 4.00-4.16 (m, 2H), 5.71 (s, 2H).
Step 2) tert-butyl 4-(2-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methoxy)-2-oxoethyl)piperidine-1-carboxylate
The title compound was prepared according to the procedure as described in Example 14 Step 2 using N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.1 g, 0.2 mmol) in DMF (0.9 mL), KOH (23 mg, 0.4 mmol) and a solution of tert-butyl 4-(2-(chloromethoxy)-2-oxoethyl)piperidine-1-carboxylate (60 mg, 0.2 mmol) in DMF (100 μL). The crude product was purified by a silica gel column chromatography (PE/EtOAc (v/v)=3/1 to 2/1) to afford the title compound as a white solid (86 mg, 56.6%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, neg. ion) m/z 742.4 [M−H]−;
1H NMR (400 MHz, CDCl3) δ 1.02-1.15 (m, 5H), 1.43 (s, 9H), 1.52-1.60 (m, 2H), 1.70-1.87 (m, 3H), 2.26-2.30 (d, J=7.0 Hz, 2H), 2.58-2.70 (m, 2H), 3.10-3.15 (m, 2H), 3.96-4.06 (m, 2H), 6.28 (s, 2H), 7.03-7.10 (m, 1H), 7.46-7.50 (m, 2H), 7.58-7.64 (m, 2H), 7.69-7.77 (m, 1H), 7.83 (s, 1H), 8.66-8.70 (d, J=2.2 Hz, 1H), 8.84-8.87 (d, J=2.2 Hz, 1H).
Step 3) (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-(piperidin-4-yl)acetate hydrochloride
A mixture of tert-butyl 4-(2-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methoxy)-2-oxoethyl) piperidine-1-carboxylate (78 mg, 0.1 mmol) and a saturated solution of HCl in EtOAc (2 mL) was stirred at rt for 2 h. The mixture was concentrated in vacuo to afford the title compound as a white solid (75 mg, 100%). The title compound was characterized by LCMS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 645.2 [M+H]+;
1H NMR (400 MHz, d6-DMSO) δ 0.96-1.00 (t, J=7.4 Hz, 3H), 1.30-1.40 (m, 2H), 1.71-1.80 (m, 4H), 1.87-1.97 (m, 1H), 2.30-2.35 (d, J=7.0 Hz, 2H), 2.75-2.87 (m, 2H), 3.13-3.22 (m, 4H), 6.31 (s, 2H), 7.31-7.38 (m, 1H), 7.58-7.70 (m, 3H), 7.79-7.85 (m, 2H), 8.48 (s, 1H), 8.67-8.70 (m, 2H), 8.80-8.82 (d, J=2.2 Hz, 1H), 9.84 (s, 1H).
Example 18
(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-amino-3-methylbutanoate hydrochloride




Step 1) chloromethyl 2-((tert-butoxycarbonyl)amino)-3-methylbutanoate
The title compound was prepared according to the procedure as described in Example 17 Step 1 using 2-((tert-butoxycarbonyl)amino)-3-methylbutanoic acid (1 g, 4.6 mmol) in DCM (25 mL) and H2O (25 mL), NaHCO3 (1.55 g, 18.4 mmol), TBAHSO4 (156 mg, 0.46 mmol) and a solution of chloromethyl sulfochloridate (559 μL, 5.5 mmol) in DCM (5 mL). The title compound was obtained as colorless oil (877 mg, 71.8%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 0.92-0.94 (d, J=6.9 Hz, 3H), 0.99-1.01 (d, J=6.9 Hz, 3H), 1.45 (s, 9H), 2.17-2.23 (m, 1H), 4.25-4.29 (m, 1H), 5.02-5.04 (d, J=8.3 Hz, 1H), 5.62-5.64 (d, J=6.0 Hz, 2H), 5.87-5.89 (d, J=6.0 Hz, 1H).
Step 2) ((tert-butoxycarbonyl)amino)-3-methylbutanoate carbon 1 amino-3-methylbutanoate
The title compound was prepared according to the procedure as described in Example 14 Step 2 using N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.1 g, 0.2 mmol) in DMF (0.9 mL), KOH (23 mg, 0.4 mmol), a solution of chloromethyl 2-((tert-butoxycarbonyl)amino)-3-methylbutanoate (54 mg, 0.2 mmol) in DMF (100 μL). The title compound was purified by a silica gel column chromatography (PE/EtOAc (v/v)=4/1 to 3/1) and was obtained as a white solid (82 mg, 55.8%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, neg. ion) m/z 716.5 [M−H]−;
1H NMR (400 MHz, CDCl3) δ 0.70-0.77 (d, J=6.7 Hz, 3H), 0.81-0.85 (d, J=6.5 Hz, 3H), 1.03-1.10 (t, J=7.4 Hz, 3H), 1.39 (s, 9H), 1.85-1.96 (m, 2H), 1.98-2.10 (m, 1H), 3.09-3.15 (m, 2H), 4.17-4.23 (m, 1H), 4.89-4.95 (m, 1H), 6.26-6.40 (dd, J=10.6 Hz, 35.8 Hz, 2H), 6.75 (br, 1H), 7.03-7.10 (m, 1H), 7.46-7.51 (m, 2H), 7.58-7.64 (m, 2H), 7.70-7.77 (m, 1H), 7.83 (s, 1H), 8.66-8.70 (d, J=2.2 Hz, 1H), 8.84-8.87 (d, J=2.1 Hz, 1H).
Step 3) (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-amino-3-methylbutanoate hydrochloride
The title compound was prepared according to the procedure as described in Example 17 Step 3 using (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl2-((tert-butoxycarbonyl)amino)-3-methyl butanoate (78 mg, 0.11 mmol) and a saturated solution of HCl in EtOAc (2 mL). The title compound was obtained as a white solid (40 mg, 54%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 619.2 [M+H]+;
1H NMR (400 MHz, d6-DMSO) δ 0.80-0.90 (t, J=6.9 Hz, 6H), 0.95-1.01 (d, J=7.4 Hz, 3H), 1.71-1.82 (m, 2H), 2.05-2.15 (m, 1H), 3.12-3.20 (m, 2H), 3.94-3.98 (d, J=4.5 Hz, 1H), 6.40-6.57 (dd, J=10.6, 35.4 Hz, 2H), 7.30-7.39 (m, 1H), 7.58-7.70 (m, 3H), 7.78-7.87 (d, J=8.5 Hz, 2H), 8.53 (s, 2H), 8.68 (s, 1H), 8.79-8.85 (d, J=2.1 Hz, 1H).
Example 19
(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-aminoacetate hydrochloride




Step 1) chloromethyl 2-((tert-butoxycarbonyl)amino)acetate
The title compound was prepared according to the procedure as described in Example 17 Step 1 using 2-((tert-butoxycarbonyl)amino)acetic acid (1 g, 5.7 mmol) in DCM (25 mL) and H2O (25 mL), NaHCO3 (1.92 g, 22.8 mmol), TBAHSO4 (194 mg, 0.57 mmol) and a solution of chloromethyl sulfochloridate (693 mL, 6.8 mmol) in DCM (5 mL). The title compound was obtained as colorless oil (911 mg, 71.3%).
Step 2) (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]-pyridin-1-yl-butoxycarbonyl)amino)acetate
The title compound was prepared according to the procedure as described in Example 14 Step 2 using N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.1 g, 0.2 mmol) in DMF (0.9 mL), KOH (23 mg, 0.4 mmol), a solution of chloromethyl 2-((tert-butoxycarbonyl)amino)acetate (46 mg, 0.2 mmol) in DMF (100 mL). The title compound was purified by a silica gel column chromatography (PE/EtOAc (v/v)=4/1 to 3/1) and was obtained as a white solid (46 mg, 33.3%). The title compound was characterized by
LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 677.1 [M+H]+;
1H NMR (400 MHz, d6-DMSO) δ 1.00-1.06 (t, J=7.4 Hz, 3H), 1.34 (s, 9H), 1.77-1.87 (m, 2H), 3.17-3.24 (m, 2H), 3.72-3.77 (d, J=6.1 Hz, 2H), 6.38 (s, 2H), 7.26-7.32 (t, J=6.1 Hz, 1H), 7.35-7.42 (m, 1H), 7.63-7.73 (m, 3H), 7.85-7.90 (d, J=8.6 Hz, 2H), 8.55 (s, 1H), 8.73 (s, 1H), 8.85-8.88 (d, J=2.2 Hz, 1H), 9.85-9.90 (br, 1H).
Step 3) (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-aminoacetate hydrochloride
The title compound was prepared according to the procedure as described in Example 17 Step 3 using (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-((tert-butoxycarbonyl)amino)acetate (46 mg, 0.07 mmol) and a saturated solution of HCl in EtOAc (2 mL). The title compound was obtained as a white solid (36 mg, 83.7%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 577.1 [M+H]+;
1H NMR (400 MHz, d6-DMSO) δ 0.95-1.00 (d, J=7.4 Hz, 3H), 1.70-1.80 (m, 2H), 3.12-3.18 (m, 2H), 3.85 (s, 1H), 6.44 (s, 2H), 7.30-7.37 (m, 1H), 7.58-7.72 (m, 3H), 7.79-7.84 (m, 2H), 8.50-8.82 (m, 2H), 8.51 (s, 1H), 8.66 (s, 1H), 8.80-8.82 (d, J=2.2 Hz, 1H), 9.84 (s, 1H).
Example 20
(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-(methylamino)acetate hydrochloride




Step 1 chloromethyl 2-((tert-butoxycarbonyl)methyl)amino acetate
The title compound was prepared according to the procedure as described in Example 17 Step 1 using 2-((tert-butoxycarbonyl)(methyl)amino)acetic acid (1 g, 5.29 mmol) in DCM (25 mL) and H2O (25 mL), NaHCO3 (1.78 g, 21.2 mmol), TBAHSO4 (0.18 g, 0.53 mmol) and a solution of chloromethyl sulfochloridate (638 μL, 6.35 mmol) in DCM (5 mL). The title compound was obtained as colorless oil (0.87 g, 69.4%).
1H NMR (400 MHz, CDCl3) δ 1.43 (s, 9H), 2.94-2.96 (d, J=5.0 Hz, 1H), 3.97-4.05 (d, J=33.9 Hz, 2H), 5.74-5.75 (d, J=4.0 Hz, 1H).
Step 2) ((tert-butoxycarbonyl)(carbonylmethyl)amino acetate
The title compound was prepared according to the procedure as described in Example 14 Step 2 using N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.3 g, 0.61 mmol) in DMF (2.5 mL), KOH (69 mg, 1.23 mmol), a solution of chloromethyl 2-((tert-butoxycarbonyl)(methyl)amino)acetate (145 mg, 0.61 mmol) in DMF (0.5 mL). The crude product was purified by a silica gel column chromatography (PE/EtOAc (v/v)=2/1) to afford the title compound as colorless oil (0.33 g, 77.9%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, neg. ion) m/z 689.4 [M−H]−;
1H NMR (400 MHz, CDCl3) δ 0.88-0.94 (t, J=7.1 Hz, 3H), 1.43 (s, 9H), 1.89-1.91 (m, 2H), 2.89 (s, 3H), 3.08-3.13 (m, 2H), 3.91-4.00 (d, J=32.7 Hz, 2H), 6.33 (s, 2H), 6.78 (br, 1H), 7.03-7.10 (m, 1H), 7.47-7.50 (m, 2H), 7.59-7.62 (m, 2H), 7.70-7.75 (m, 1H), 7.82-7.86 (m, 1H), 8.66-8.68 (d, J=1.6 Hz, 1H), 8.85-8.87 (d, J=1.6 Hz, 1H).
Step 3) (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-(methylamino)acetate hydrochloride
A mixture of (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-((tert-butoxycarbonyl)(methyl)amino)acetate (0.33 g, 0.48 mmol) and a saturated solution of HCl in EtOAc (3 mL) was stirred at rt for 3 h. The mixture was filtered to afford the title compound as a white solid (168 mg, 56%). The title compound was characterized by LCMS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 591.1 [M+H]+;
LC-MS (ESI, neg. ion) m/z 588.5 [M−H]−;
1H NMR (400 MHz, d6-DMSO) δ 0.96-1.00 (t, J=7.4 Hz, 3H), 1.73-1.79 (m, 2H), 2.49-2.51 (m, 3H), 3.13-3.17 (m, 2H), 4.02 (s, 2H), 6.45 (s, 2H), 7.32-7.36 (m, 1H), 7.59-7.62 (dd, J=2.0, 6.6 Hz, 2H), 7.62-7.67 (m, 1H), 7.80-7.83 (dd, J=2.0, 6.6 Hz, 2H), 8.51 (s, 1H), 8.67 (br, 1H), 8.80-8.81 (d, J=2.2 Hz, 1H), 8.99 (br, 1H).
Example 21
(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-amino-2-methylpropanoate hydrochloride




Step 1) chloromethyl 2-((tert-butoxycarbonyl)amino)-2-methylpropanoate
The title compound was prepared according to the procedure as described in Example 17 Step 1 using 2-((tert-butoxycarbonyl)amino)-2-methylpropanoic acid (1.07 g, 5.29 mmol) in DCM (25 mL) and H2O (25 mL), NaHCO3 (1.78 g, 21.2 mmol), TBAHSO4 (0.18 g, 0.53 mmol) and a solution of chloromethyl sulfochloridate (638 μL, 6.35 mmol) in DCM (5 mL). The title compound was obtained as a white solid (1.16 g, 87.7%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.43 (s, 9H), 1.51 (s, 6H), 4.90 (m, 1H), 5.74-5.75 (d, J=4.0 Hz, 1H).
Step 2) (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-((tert-butoxycarbonyl)amino)-2-methylpropanoate
The title compound was prepared according to the procedure as described in Example 14 Step 2 using N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.3 g, 0.61 mmol) in DMF (2.5 mL), KOH (69 mg, 0.4 mmol), and a solution of chloromethyl 2-((tert-butoxycarbonyl)amino)-2-methylpropanoate (158 mg, 0.61 mmol) in DMF (0.5 mL). The title compound was purified by a silica gel columnchromatography (PE/EtOAc (v/v)=4/1 to 3/1) and was obtained as a white solid (339 mg, 78.7%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 705.2 [M+H]+;
1H NMR (400 MHz, CDCl3) δ 1.02-1.06 (t, J=7.4 Hz, 3H), 1.21 (s, 9H), 1.41 (s, 6H), 1.85-1.92 (m, 2H), 2.89 (s, 3H), 3.07-3.12 (m, 2H), 4.93 (br, 1H), 6.31 (s, 2H), 7.01-7.05 (m, 1H), 7.45-7.49 (m, 2H), 7.57-7.61 (m, 2H), 7.67-7.73 (m, 1H), 7.87 (s, 1H), 8.64-8.65 (d, J=2.2 Hz, 1H), 8.83-8.84 (d, J=2.2 Hz, 1H).
Step 3) (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-amino-2-methylpropanoate hydrochloride
A mixture of (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-((tert-butoxycarbonyl)amino)-2-methylpropanoate (339 mg, 0.48 mmol) and a saturated solution of HCl in EtOAc (3 mL) was stirred at rt for 5 h. The resulting mixture was filtered to afford the title compound as a white solid (224 mg, 72.6%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 605.2 [M+H]+;
LC-MS (ESI, neg. ion) m/z 603.3 [M−H]−;
1H NMR (400 MHz, d6-DMSO) δ 0.95-0.99 (t, J=7.4 Hz, 3H), 1.40 (s, 6H), 1.73-1.79 (m, 2H), 3.12-3.16 (m, 2H), 6.44 (s, 2H), 7.32-7.36 (m, 1H), 7.59-7.61 (dd, J=2.0 Hz, 6.6 Hz, 2H), 7.62-7.67 (m, 1H), 7.80-7.83 (dd, J=2.0, 6.6 Hz, 2H), 8.48 (br, 2H), 8.52 (s, 1H), 8.67 (br, 1H), 8.79-8.80 (d, J=2.2 Hz, 1H).
Example 22
(2S)-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl-2-amino-3-methylpentanoate hydrochloride




Step 1) (2S)-chloromethyl 2-((tert-butoxycarbonyl)amino)-3-methylpentanoate
The title compound was prepared according to the procedure as described in Example 17 Step 1 using L-2-((tert-butoxycarbonyl)amino)-3-methylpentanoic acid (1 g, 4.31 mmol) in DCM (25 mL) and H2O (25 mL), NaHCO3 (1.55 g, 18.4 mmol), TBAHSO4 (156 mg, 0.46 mmol) and a solution of chloromethyl sulfochloridate (559 μL, 5.5 mmol) in DCM (5 mL). The title compound was obtained as colorless oil (0.91 g, 76.1%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 0.91-0.95 (t, J=7.4 Hz, 3H), 0.96-0.98 (d, J=6.9 Hz, 3H), 1.24-1.29 (m, 2H), 1.44 (s, 9H), 1.87-1.93 (m, 1H), 4.29-4.33 (m, 1H), 4.97-5.00 (m, 1H), 5.61-5.63 (d, J=6.0 Hz, 1H), 5.87-5.89 (d, J=6.0 Hz, 1H).
Step 2) (2S)-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-((tert-butoxycarbonyl)amino-3-methylpentanoate
The title compound was prepared according to the procedure as described in Example 14 Step 2 using N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.3 g, 0.61 mmol) in DMF (2.5 mL), KOH (69 mg, 1.22 mmol) and a solution of chloromethyl 2-((tert-butoxycarbonyl)amino)-3-methyl pentanoate (171 mg, 0.61 mmol) in DMF (0.5 mL). The crude product was purified by a silica gel column chromatography (PE/EtOAc (v/v)=4/1 to 3/1) to afford the title compound as a white solid (288 mg, 64%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 733.1 [M+H]
1H NMR (400 MHz, CDCl3) δ 0.69-0.72 (t, J=7.3 Hz, 3H), 0.77-0.79 (d, J=7.3 Hz, 3H), 0.84-0.91 (m, 2H), 1.04-1.09 (t, J=7.4 Hz, 3H), 1.39 (s, 9H), 1.72-1.79 (m, 1H), 1.87-1.94 (m, 2H), 3.09-3.14 (m, 2H), 4.25-4.27 (m, 1H), 4.89 (br, 1H), 6.26-6.29 (d, J=10.6 Hz, 1H), 6.36-6.40 (d, J=10.6 Hz, 1H), 6.63 (br, 1H), 7.04-7.09 (m, 1H), 7.47-7.50 (dd, J=2.0, 6.6 Hz, 2H), 7.59-7.62 (dd, J=2.0, 6.6 Hz, 2H), 7.71-7.79 (td, J=5.5, 9.0 Hz, 1H), 7.83 (s, 1H), 8.67-8.68 (d, J=2.1 Hz, 1H), 8.85-8.86 (d, J=2.1 Hz, 1H).
Step 3) (2S)-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-amino-3-methylpentanoate hydrochloride
A mixture of (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-((tert-butoxycarbonyl)amino)-3-methyl pentanoate (285 mg, 0.45 mmol) and a saturated solution of HCl in EtOAc (3 mL) was stirred at 0° C. for 10 h. The resulting mixture was filtered to afford the title compound as a white solid (0.21 g, 80.8%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 633.1 [M+H]+;
1H NMR (400 MHz, d6-DMSO) δ 0.69-0.72 (t, J=7.0 Hz, 3H), 0.75-0.78 (d, J=6.1 Hz, 3H), 0.95-1.00 (t, J=7.4 Hz, 3H), 1.03-1.07 (m, 2H), 1.72-1.79 (m, 2H), 1.79-1.84 (m, 1H), 3.12-3.16 (m, 2H), 3.95-3.97 (m, 1H), 6.37-6.41 (d, J=10.6 Hz, 1H), 6.51-6.55 (d, J=10.6 Hz, 1H), 7.31-7.36 (m, 1H), 7.58-7.61 (d, J=8.2 Hz, 2H), 7.62-7.67 (m, 1H), 7.80-7.83 (d, J=8.4 Hz, 2H), 8.52 (s, 1H), 8.55-8.63 (m, 2H), 8.67 (s, 1H), 8.80-8.82 (d, J=2.2 Hz, 1H), 9.88 (br, 1H).
Example 23
(S)-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-amino-4-methylpentanoate hydrochloride




Step 1) chloromethyl 2-((tert-butoxycarbonyl)amino)-4-methylpentanoate
The title compound was prepared according to the procedure as described in Example 17 Step 1 using L-2-((tert-butoxycarbonyl)amino)-4-methylpentanoic acid (1 g, 4.31 mmol) in DCM (25 mL) and H2O (25 mL), NaHCO3 (1.55 g, 18.4 mmol), TBAHSO4 (156 mg, 0.46 mmol) and a solution of chloromethyl sulfochloridate (559 μL, 5.5 mmol) in DCM (5 mL). The title compound was obtained as colorless oil (0.98 g, 81.1%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 0.94-0.97 (dd, J=1.2, 6.5 Hz, 6H), 1.44 (s, 9H), 1.50-1.54 (m, 1H), 1.70-1.76 (m, 1H), 4.33-4.35 (m, 1H), 4.83-4.86 (m, 1H), 5.61-5.63 (d, J=6.0 Hz, 1H), 5.85-5.87 (d, J=6.0 Hz, 1H).
Step 2) (S)-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-(tert-butoxylcarbonyl)amino)-4-methylpentanoate
The title compound was prepared according to the procedure as described in Example 14 Step 2 using N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.3 g, 0.61 mmol) in DMF (2.5 mL), KOH (69 mg, 1.22 mmol), and a solution of chloromethyl 2-((tert-butoxy-carbonyl)amino)-4-methylpentanoate (179 mg, 0.61 mmol) in DMF (0.5 mL). The crude product was purified by a silica gel column chromatography (PE/EtOAc (v/v)=4/1 to 3/1) to afford the title compound as a white solid (287 mg, 62.8%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 733.1 [M+H]+;
1H NMR (400 MHz, CDCl3) δ 0.82-0.85 (d, J=6.2 Hz, 6H), 0.89-0.93 (m, 1H), 1.04-1.09 (t, J=7.4 Hz, 3H), 1.24-1.27 (m, 2H), 1.36 (s, 9H), 1.87-1.94 (m, 2H), 3.09-3.14 (m, 2H), 4.25-4.27 (m, 1H), 4.79 (br, 1H), 6.26-6.29 (d, J=10.6 Hz, 2H), 6.35-6.38 (d, J=10.6 Hz, 2H), 6.68 (br, 1H), 7.04-7.09 (m, 1H), 7.47-7.50 (dd, J=2.0, 6.6 Hz, 2H), 7.59-7.62 (dd, J=2.0, 6.6 Hz, 2H), 7.70-7.77 (td, J=5.5, 9.0 Hz, 1H), 7.83 (s, 1H), 8.67-8.68 (d, J=2.1 Hz, 1H), 8.85-8.86 (d, J=2.1 Hz, 1H).
Step 3) (S)-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl 2-amino-4-methylpentanoate hydrochloride
A solution of (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl2-((tert-butoxycarbonyl)amino)-4-methylpentanoate (284 mg, 0.39 mmol) was added in a saturated solution of HCl in EtOAc (3 mL). The reaction mixture was stirred at 0° C. for 10 h. The resulting mixture was concentrated in vacuo, and washed with PE/EtOAc (1/1) to afford the title compound as a white solid (204 mg, 78.7%).
LC-MS (ESI, pos. ion) m/z 633.1 [M+H]+;
1H NMR (400 MHz, d6-DMSO) δ 0.73-0.76 (t, J=5.4 Hz, 6H), 0.84-0.85 (m, 1H), 0.95-1.00 (t, J=7.4 Hz, 3H), 1.16-1.18 (m, 2H), 1.72-1.79 (m, 2H), 3.12-3.17 (m, 2H), 3.99-4.03 (m, 1H), 6.41-6.49 (dd, J=10.5, 19.5 Hz, 2H), 7.31-7.35 (m, 1H), 7.58-7.61 (dd, J=2.0, 6.6 Hz, 2H), 7.61-7.66 (m, 1H), 7.80-7.83 (dd, J=2.0, 6.6 Hz, 2H), 8.47 (br, 2H), 8.51 (s, 1H), 8.67 (br, 1H), 8.80-8.81 (d, J=2.2 Hz, 1H), 9.84 (s, 1H).
Example 24
1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)-benzoy)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl 2-amino-3-methylbutanoate hydrochloride




Step 1) 1-chloroethyl sulfochloridate
Neat chlorosulfonic acid (8.96 g, 76.94 mmol) was added to 1-chloroethyl chloroformate dropwise at 0° C. The resulting mixture was stirred at 0° C. 16 h, then quenched with 15 mL of DCM and 15 mL of H2O. The organic phase was washed with saturated Na2CO3 aqueous solution (20 mL), followed by brine (20 mL), dried over anhydrous Na2SO4 and concentrated in vacuo. The residue was distilled at 54° C. under reduced pressure to give the title compound as colorless oil (4.5 g, 36%). The title compound was characterized by 1H NMR and 13C NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.98-1.99 (d, J=5.8 Hz, 3H), 6.46-6.50 (q, J=5.8 Hz, 1H);
13C NMR (100 MHz, CDCl3) δ 26.3, 91.1.
Step 2) 1-chloroethyl 2-((tert-butoxycarbonyl)amino)-3-methylbutanoate
The title compound was prepared according to the procedure as described in Example 17 step 1 using 2-((tert-butoxycarbonyl)amino)-3-methylbutanoic acid (3 g, 13.81 mmol) in DCM (75 mL) and H2O (75 mL), NaHCO3 (4.64 g, 55.23 mmol), TBAHSO4 (469 mg, 1.38 mmol) and a solution of 1-chloroethyl sulfochloridate (2.97 g, 16.57 mmol) in DCM (15 mL). The crude product was purified by a silica gel column chromatography (PE/EtOAc (v/v)=100/1 to 50/1) to afford the title compound as a white solid (3 g, 78%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 0.90-0.92 (m, 3H), 0.97-0.99 (m, 3H), 1.45 (s, 9H), 1.79-1.81 (m, 3H), 2.14-2.19 (m, 1H), 4.22-4.25 (m, 1H), 4.99-5.01 (m, 1H), 6.53-6.57 (dd, J=5.7, 11 Hz, 1H).
Step 3) 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl2-((tert-butoxycarbonyl)amino)-3-methylbutanoate
The title compound was prepared according to the procedure as described in Example 14 step 2 using N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.3 g, 0.61 mmol) in anhydrous DMF (2.5 mL), KOH (69 mg, 1.22 mmol) and a solution of 1-chloroethyl 2-((tert-butoxycarbonyl)amino)-3-methylbutanoate (172 mg, 0.61 mmol) in anhydrous DMF (0.5 mL). The crude product was purified by a silica gel column chromatography (PE/EtOAc (v/v)=4/1 to 3/1) to afford the title compound as orange oil (0.29 g, 65%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 0.82-0.92 (m, 6H), 0.92-0.94 (t, J=7.4 Hz, 3H), 1.25 (s, 9H), 1.89-1.96 (m, 5H), 2.10-2.13 (m, 1H), 3.11-3.12 (m, 2H), 4.17-4.21 (m, 1H), 6.95-7.13 (m, 2H), 7.40-7.47 (m, 2H), 7.58-7.60 (m, 2H), 7.80-7.82 (m, 1H), 8.01 (s, 1H), 8.63-8.65 (m, 1H), 8.80-8.83 (m, 1H).
Step 4) 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)-benzoy)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl 2-amino-3-methylbutanoate hydrochloride
A mixture of 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl 2-((tert-butoxycarbonyl)amino)-3-methylbutanoate (0.29 g, 0.39 mmol) and a saturated solution of HCl in EtOAc (4 mL) was stirred at rt for 2 h. The mixture was filtered to give the title compound as a white solid (0.15 g, 57%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 634.1 [(M+H)+−HCl];
1H NMR (400 MHz, CDCl3) δ 0.85-0.89 (m, 6H), 0.98-0.99 (t, J=7.4 Hz, 3H), 1.76-1.89 (m, 3H), 1.92-2.03 (m, 3H), 3.00-3.13 (m, 2H), 4.00-4.05 (m, 1H), 6.95-7.13 (m, 2H), 7.40-7.47 (m, 2H), 7.58-7.60 (m, 2H), 7.80-7.82 (m, 1H), 8.01 (s, 1H), 8.63-8.65 (m, 1H), 8.80-8.83 (m, 1H).
Example 25
sodium (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl phosphate




Step 1) di-tert-butyl(chloromethyl) phosphate
The title compound was prepared according to the procedure as described in Example 17 Step 1 using potassium di-tert-butylphosphate (1 g, 4.03 mmol) in DCM (20 mL) and H2O (20 mL), NaHCO3 (1.34 g, 16.1 mmol), TBAHSO4 (136 mg, 0.4 mmol) and a solution of chloromethyl sulfochloridate (452 μL, 4.41 mmol) in DCM (5 mL). The title compound was obtained as colorless oil (1.02 g, 95.3%). The title compound was characterized by 1H NMR and 13C NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.51 (s, 18H), 5.62-5.66 (d, J=14.9 Hz, 2H);
13C NMR (100 MHz, CDCl3) δ 29.9, 73.3, 84.1.
Step 2) di-tert-butyl((5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl)phosphate
To a solution of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.4 g, 0.82 mmol) in DMF (3.5 mL) was added KOH (91 mg, 1.63 mmol). The reaction was stirred at 10° C. for 20 min, followed by the dropwise addition of a solution of di-tert-butyl(chloromethyl) phosphate (0.21 g, 0.82 mmol) in DMF (1 mL) over 30 min. The reaction was stirred further at 10° C. for 16 h, then diluted with EtOAc (100 mL). The mixture was washed with H2O (100 mL) and extracted with DCM (100 mL×2). The combined organic phases were concentrated in vacuo and the residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=3/1) to give the title compound ascolorless oil (0.28 g, 47.2%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 712.2 [M+H]+;
1H NMR (400 MHz, CDCl3) δ 1.03-1.07 (t, J=7.4 Hz, 3H), 1.35 (s, 18H), 1.84-1.94 (m, 2H), 3.07-3.11 (m, 2H), 6.13-6.17 (d, J=13.0 Hz, 2H), 7.01-7.06 (m, 1H), 7.46-7.49 (d, J=8.4 Hz, 2H), 7.58-7.60 (d, J=8.4 Hz, 2H), 7.67-7.74 (td, J=8.9, 5.6 Hz, 1H), 7.96 (s, 1H), 8.67-8.68 (d, J=2.1 Hz, 1H), 8.87-8.88 (d, J=2.0 Hz, 1H).
Step 3) (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl dihydrogen phosphate
To a solution of di-tert-butyl((5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propyl sulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl) phosphate (1 g, 1.4 mmol) in DCM (25 mL) was added TFA (1.6 mL, 2.4 g, 21 mmol). The reaction was stirred at rt for 2 h, and then concentrated in vacuo. The residue was washed with 20 mL of EtOAc to give the title compound as a white solid (0.56 g, 66.5%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 600.0 [M+H]+;
1H NMR (400 MHz, d6-DMSO) δ 0.94-0.98 (t, J=7.4 Hz, 3H), 1.70-1.77 (m, 2H), 3.10-3.14 (m, 2H), 6.03-6.06 (d, J=9.6 Hz, 2H), 7.29-7.33 (m, 1H), 7.57-7.60 (d, J=8.5 Hz, 2H), 7.60-7.65 (m, 1H), 7.80-7.82 (d, J=8.5 Hz, 2H), 8.42-8.43 (m, 1H), 8.65 (s, 1H), 8.79-8.80 (d, J=2.1 Hz, 1H), 9.82 (br, 1H).
Step 4) sodium (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl phosphate
To a solution of (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl dihydrogen phosphate (396 mg, 0.66 mmol) in THF (20 mL) was added dropwise a solution of NaOH (58 mg, 1.45 mmol) in 0.3 mL of H2O and 4 mL of THF at rt. The reaction was stirred at rt for 4 h, and then filtered to give the title compound as a white solid (401 mg, 94.4%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 600.0 [M+H]+;
1H NMR (400 MHz, D2O) δ 0.82-0.86 (t, J=7.4 Hz, 3H), 1.62-1.68 (m, 2H), 2.88-2.92 (m, 2H), 5.79-5.82 (d, J=9.6 Hz, 2H), 6.96-7.01 (m, 1H), 7.27-7.30 (d, J=8.5 Hz, 2H), 7.31-7.37 (td, J=9.3, 6.0 Hz, 1H), 7.41-7.43 (d, J=8.5 Hz, 2H), 8.34-8.35 (m, 2H), 8.46-8.47 (d, J=2.0 Hz, 1H).
Example 26
(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl ethyl carbonate




Step 1) chloromethyl ethyl carbonate
To a solution of chloromethyl chloroformate (10 g, 78.2 mmol) in DCM (200 mL) was added a mixture of ethanol (5 mL, 85.9 mmol) and pyridine (7.5 mL, 93.2 mmol) in DCM (50 mL) at −20° C. The reaction was stirred at 0° C. for 1 h, then at rt for another 2 h. The resulting mixture was washed with 1N HCl solution (300 mL), followed by brine (200 mL). The organic phase was dried over anhydrous Na2SO4 and concentrated in vacuo to give the title compound as colorless oil (8.75 g, 81.1%). The compound was used in the next step without further purification. The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.32-1.36 (t, J=7.2 Hz, 3H), 4.26-4.31 (m, J=7.2 Hz, 2H), 5.72 (s, 2H).
Step 2) (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl ethyl carbonate
To a mixture of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.1 g, 0.2 mmol) and Et3N (114 μL, 0.8 mmol) in DMF (0.6 mL) was added a solution of TBAB (132 mg, 0.4 mmol) in DMF (0.2 mL). The reaction was stirred at 10° C. for 30 min, followed by dropwise addition of a solution of chloromethyl ethyl carbonate (34 mg, 0.25 mmol) in DMF (0.2 mL). The mixture was stirred at 10° C. for another 3 h, then diluted with EtOAc (40 mL) and filtered. The filtrate was concentrated in vacuo and the residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=2/1) to afford the title compound as a white solid (85 mg, 73%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 592.0 [M+H]+;
1H NMR (400 MHz, CDCl3) δ 1.05-1.10 (t, J=7.4 Hz, 3H), 1.25-1.32 (t, J=7.2 Hz, 3H), 1.85-1.97 (m, 2H), 3.09-3.16 (m, 2H), 4.18-4.25 (q, J=7.1 Hz, 2H), 6.29 (s, 2H), 6.51 (s, 1H), 7.04-7.10 (m, 1H), 7.46-7.51 (d, J=8.5 Hz, 2H), 7.57-7.63 (d, J=8.5 Hz, 2H), 7.70-7.77 (m, 1H), 7.86 (s, 1H), 8.67-8.70 (d, J=2.1 Hz, 1H), 8.83-8.85 (d, J=2.1 Hz, 1H).
Example 27
(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl isopropyl carbonate




Step 1) chloromethyl isopropyl carbonate
The title compound was prepared according to the procedure as described in Example 26 step 1 using a solution of chloromethyl chloroformate (10 g, 78.2 mmol) in DCM (200 mL), a mixture of i-PrOH (6.6 mL, 86.2 mmol) and pyridine (7.5 mL, 93.2 mmol) in DCM (50 mL). The title compound was obtained as colorless oil (9.63 g, 81.0%) and was used in the next step without further purification. The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.33 (s, 3H), 1.34 (s, 3H), 4.92-4.99 (m, J=6.3 Hz, 1H), 5.72 (s, 3H).
Step 2) (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl isopropyl carbonate
To a mixture of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.2 g, 0.41 mmol) and KOH (46 mg, 0.82 mmol) in DMF (1.5 mL) was added a solution of chloromethyl isopropyl carbonate (62 mg, 0.41 mmol) in DMF (0.5 mL) over 30 min. The reaction mixture was stirred at 10° C. for 3 h, then diluted with EtOAc (40 mL) and filtered. The filtrate was concentrated in vacuo and the residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=2/1) to afford the title compound as a white solid (105 mg, 43%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 606.0 [M+H]+;
1H NMR (400 MHz, CDCl3) δ 1.04-1.10 (t, J=7.4 Hz, 3H), 1.20-1.35 (m, 6H), 1.85-1.95 (m, 2H), 3.09-3.15 (m, 2H), 4.84-4.92 (m, 1H), 6.28 (s, 2H), 6.56 (s, 1H), 7.03-7.10 (m, 1H), 7.46-7.51 (d, J=8.5 Hz, 2H), 7.57-7.63 (d, J=8.5 Hz, 2H), 7.69-7.77 (m, 1H), 7.87 (s, 1H), 8.66-8.70 (d, J=2.1 Hz, 1H), 8.83-8.87 (d, J=2.1 Hz, 1H).
Example 28
1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl ethyl carbonate




Step 1) 1-chloroethyl ethyl carbonate
The title compound was prepared according to the procedure as described in Example 26 step 1 using a solution of 1-chloroethyl chloroformate (10 g, 69.9 mmol) in DCM (200 mL), a mixture of ethanol (4.5 mL, 77.2 mmol) and pyridine (6.8 mL, 84.5 mmol) in DCM (50 mL). The title compound was obtained as colorless oil (9.32 g, 87.7%) and was used in the next step without further purification. The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.32-1.36 (t, J=7.1 Hz, 3H), 1.82-1.84 (d, J=5.8 Hz, 3H), 4.24-4.30 (q, J=7.1 Hz, 2H), 6.41-6.46 (q, J=5.8 Hz, 1H).
Step 2) 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl ethyl carbonate
To a solution of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.1 g, 0.2 mmol) in DMF (1 mL) was added K2CO3 (85 mg, 0.6 mmol). The reaction was stirred at rt for 10 min, followed by the addition of TBAB (132 mg, 0.4 mmol) and a solution of 1-chloroethyl ethyl carbonate (35 mg, 0.25 mmol) in DMF (100 μA). The mixture was stirred at rt for 16 h, then diluted with EtOAc (40 mL) and filtered. The filtrate was concentrated in vacuo and the residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=4/1) to afford the title compound as a white solid (61 mg, 50%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, neg. ion) m/z 603.5 [M−H]−;
1H NMR (400 MHz, CDCl3) δ 1.04-1.10 (t, J=7.4 Hz, 3H), 1.25-1.33 (t, J=7.1 Hz, 3H), 1.85-1.93 (m, 2H), 1.93-2.00 (d, J=6.3 Hz, 3H), 3.09-3.17 (m, 2H), 4.12-4.27 (m, 2H), 6.55 (br, 1H), 7.04-7.10 (m, 1H), 7.29-7.36 (q, J=6.3 Hz, 1H), 7.45-7.50 (m, 2H), 7.57-7.63 (m, 2H), 7.68-7.77 (m, 1H), 7.79 (s, 1H), 8.67-8.70 (d, J=2.2 Hz, 1H), 8.80-8.84 (d, J=2.1 Hz, 1H).
Example 29
1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl cyclohexyl carbonate




Step 1) 1-chloroethyl cyclohexyl carbonate
The title compound was prepared according to the procedure as described in Example 26 step 1 using a solution of 1-chloroethyl carbonochloridate (5 g, 35 mmol) in DCM (100 mL), cyclohexanol (3.9 g, 38.5 mmol) and pyridine (3.3 g, 42 mmol). The title compound was obtained as colorless oil (7 g, 96.9%) and was used in the next step without further purification. The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.28-1.25 (m, 1H), 1.30-1.42 (m, 2H), 1.48-1.58 (m, 3H), 1.74-1.78 (m, 2H), 1.82-1.83 (d, J=5.8 Hz, 3H), 1.91-1.96 (m, 2H), 4.66-4.72 (m, 1H), 6.41-6.45 (m, 1H).
Step 2) 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl cyclohexyl carbonate
To a solution of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (5 g, 10.2 mmol) in THF (36 mL) was added KOH (1.1 g, 20.4 mmol). The reaction was stirred at rt for 0.5 h, followed by the addition of a solution of 1-chloroethyl cyclohexyl carbonate (2.3 g, 11.2 mmol) in THF (9 mL). The mixture was refluxed for 16 h, then cooled to rtand diluted with THF (50 mL). The resulting mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=8/1) to give the title compound as a white solid (3.49 g, 51.9%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, neg. ion) m/z 657.5 [M−H]−;
1H NMR (400 MHz, CDCl3) δ 1.05-1.08 (t, J=7.4 Hz, 3H), 1.20-1.58 (m, 8H), 1.72 (m, 2H), 1.89-1.94 (m, 5H), 3.10-3.14 (m, 2H), 4.59 (m, 1H), 6.40-6.50 (d, J=1.7 Hz, 1H), 7.04-7.09 (m, 1H), 7.30-7.33 (m, 1H), 7.46-7.48 (m, 1H), 7.58-7.60 (m, 1H), 7.71-7.74 (m, 1H), 7.79 (s, 1H), 8.67 (d, J=2.2 Hz, 1H), 8.81-8.82 (d, J=2.0 Hz, 1H).
Example 30
1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl isopropyl carbonate




Step 1) 1-chloroethyl isopropyl carbonate
To a solution of 1-chloroethyl carbonochloridate (10 g, 69.9 mmol) in DCM (200 mL) was added a solution of isopropanol (5.9 mL, 76.9 mmol) and pyridine (6.75 mL, 83.9 mmol) in DCM (50 mL) at −20° C. The reaction was stirred at 0° C. for 2 h, then at rt for another 2 h. The mixture was washed with 1N HCl solution (250 mL×2), followed by brine (250 mL). The organic phase was dried over anhydrous Na2SO4 and concentrated in vacuo to give the title compound as colorless oil (9.6 g, 82.4%). The product was used in the next step without further purification. The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 1.31-1.34 (m, 6H), 1.81-1.82 (d, J=5.84 Hz, 3H), 4.88-4.98 (m, 1H), 6.41-6.45 (t, J=5.84 Hz, 1H).
Step 2) 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl isopropyl carbonate
The title compound was prepared according to the procedure as described in Example 26 step 2 using N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.3 g, 0.61 mmol), KOH (69 mg, 1.23 mmol) in anhydrous DMF (2 mL), a solution of TBAB (396 mg, 1.23 mmol) in anhydrous DMF (0.5 mL) and a solution of 1-chloroethyl isopropyl carbonate (101 mg, 0.61 mmol) in anhydrous DMF (0.5 mL). The crude product was purified by a silica gel column chromatography (PE/EtOAc (v/v)=5/1), and recrystallized in methanol to afford the title compound as a white solid (42 mg, 11.1%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, neg. ion) m/z 617.4 [M−H]−;
1H NMR (400 MHz, CDCl3) δ 1.04-1.08 (t, J=7.4 Hz, 3H), 1.22-1.29 (m, 6H), 1.87-1.94 (m, 5H), 3.10-3.14 (m, 2H), 4.84-4.87 (m, 1H), 6.59 (br, 1H), 7.04-7.09 (m, 1H), 7.30-7.34 (t, J=6.2 Hz, 1H), 7.46-7.48 (d, J=8.5 Hz, 2H), 7.58-7.60 (d, J=8.5 Hz, 2H), 7.69-7.75 (td, J=5.5, 9.0 Hz, 1H), 7.79 (s, 1H), 8.67-8.68 (d, J=2.2 Hz, 1H), 8.81-8.82 (d, J=2.0 Hz, 1H).
Example 31
(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl phenyl carbonate




Step 1) chloromethyl phenyl carbonate
To a solution of chloromethyl chloroformate (1 g, 7.8 mmol) in DCM (40 mL) was added a mixture of phenol (0.73 g, 7.8 mmol) and pyridine (750 μL, 9.3 mmol) in DCM (10 mL) at −20° C. over 20 min. The reaction was stirred at 0° C. for 4 h, then at rt for another 1 h. The mixture was washed with 1N HCl solution (50 mL), followed by brine (50 mL). The organic phase was dried over anhydrous Na2SO4 and concentrated in vacuo. The residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=100/1) to give the title compound as colorless oil (847 mg, 58.2%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3) δ 5.84 (s, 2H), 7.22-7.27 (d, J=8.2 Hz, 2H), 7.28-7.34 (t, J=7.4 Hz, 1H), 7.40-7.49 (t, J=7.8 Hz, 2H).
Step 2) (5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)methyl phenyl carbonate
To a mixture of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (0.1 g, 0.2 mmol), TBAB (132 mg, 0.4 mmol) and Et3N (114 μL, 0.8 mmol) in DMF (1 mL) was added chloromethyl phenyl carbonate (46 mg, 0.25 mmol). The reaction was stirred at rt for 12 h, then diluted with EtOAc (40 mL) and filtered. The filtrate was concentrated in vacuo and the residue was purified by a silica gel column chromatography (PE/EtOAc (v/v)=4/1) to afford the title compound as a white solid (10 mg, 7.7%). The title compound was characterized by LCMS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z 640.0 [M+H]+;
1H NMR (400 MHz, CDCl3) δ 0.98-1.12 (t, J=7.4 Hz, 3H), 1.80-1.90 (m, 2H), 3.02-3.09 (m, 2H), 6.40 (s, 2H), 6.51 (s, 1H), 7.02-7.08 (m, 1H), 7.11-7.16 (d, J=7.6 Hz, 2H), 7.25-7.30 (m, 1H), 7.37-7.42 (m, 2H), 7.46-7.52 (d, J=8.4 Hz, 2H), 7.57-7.64 (d, J=8.4 Hz, 2H), 7.67-7.75 (m, 1H), 7.88 (s, 1H), 8.70-8.73 (d, J=2.1 Hz, 1H), 8.87-8.90 (d, J=2.1 Hz, 1H).
Example 32
sodium 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl phosphate




Step 1) 1-chloroethyl sulfochloridate
To a solution of 1-Chloroethyl chloroformate (54.4 mL, 504 mmol) was added chlorosulfonic acid (49.0 mL, 729 mmol) dropwise at 0° C. over 40 min. The mixture was stirred at 0° C. for 2 h, and then 5° C. for another 10 min. To the reaction mixture were added CH2Cl2 (500 mL) and then ice (20 g) slowly. The mixture was washed with saturate aqueous NaHCO3 (400 mL) and brine (400 mL), dried over anhydrous Na2SO4, filtered, and concentrated to give light yellow oil (54.2 g, 60%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3): δ 1.98-2.00 (d, J=5.8 Hz, 3H), δ 6.46-6.51 (q, J=5.8 Hz, 1H).
Step 2) Di-tert-butyl 1-chloroethyl phosphate
To a solution of potassium di-tert-butylphosphate (33 g, 133 mmol) in CH2Cl2/H2O (600 mL/600 mL) were added NaHCO3 (44.6 g, 531 mmol) and TBAHSO4 (4.52 g, 13.3 mmol), and then the mixture was stirred at 0° C. for 10 min. Then a solution of 1-chloroethyl sulfochloridate (26 g, 146 mmol) in CH2Cl2 (100 mL) was added dropwise over 30 min to the reaction mixture. The mixture was stirred further at room temperature for 20 h. The organic layer was separated, and then washed with water (300 mL×3) and brine (300 mL). Then it was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give light yellow oil (14.3 g, 40%). The title compound was characterized by 1H NMR as shown below:
1H NMR (400 MHz, CDCl3): δ 1.52 (s, 18H), 1.81 (d, J=5.6 Hz, 3H), 6.17-6.24 (m, 1H).
Step 3) di-tert-butyl 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl phosphate
To a solution of N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide (9.8 g, 20 mmol) in DMF (120 mL) were added KOH (4.48 g, 80 mmol) and KI (0.33 g, 2 mmol). Then a solution of di-tert-butyl 1-chloroethyl phosphate (10.9 g, 40 mmol) in DMF (20 mL) was added dropwise to the above reaction mixture at rt over 5 min under N2 protection and then the reaction mixture was stirred at 50° C. for 6 h, followed by being diluted with ethyl acetate (700 mL). The mixture was washed with water (400 mL×3) and brine (400 mL). The organic layer was concentrated, and then CH2Cl2 (20 mL) was added. The solid was collected through filtration in vacuo, and then the filter was concentrated to give a brown solid (31 g, 106%). The title compound was characterized by LC-MS as shown below:
LC-MS (ESI, pos. ion) m/z: 724.2 [M−1].
Step 4) 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl dihydrogen phosphate
To the solution of di-tert-butyl 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl phosphate (20 g, crude) in EA (30 mL) was added phosphate (85%, 90 mL) and the mixture was stirred at room temperature for 2 min. Then the reaction mixture was quenched by adding water (200 mL), and then extracted with EtOAc (300 mL×3). The combined organic layers were concentrated in vacuo to give a brown solid (19.5 g, 74%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z: 612.1 [M−1];
1H NMR (400 MHz, DMSO-d6): δ 0.95 (t, J=7.4 Hz, 3H), 1.65-1.81 (m, 5H), 3.11 (t, J=7.4 Hz, 2H), 6.75-6.87 (m, 1H), 7.28 (t, J=8.8 Hz, 1H), 7.57 (d, J=8.3 Hz, 2H), 7.51-7.72 (m, 1H), 7.77 (d, J=8.3 Hz, 2H), 8.31-8.35 (m, 1H), 8.57-8.61 (m, 1H), 8.69-8.74 (m, 1H), 9.85 (br, 1H).
Step 5) Sodium 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl phosphate
To the solution of 1-(5-(4-chlorophenyl)-3-(2,6-difluoro-3-(propylsulfonamido)benzoyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl dihydrogen phosphate (4 g, 6.5 mmol) in EA/THF (30 mL, 2/1) was added NaOH (0.57 g, 14.3 mmol) in H2O (1 mL) at rt. Then the mixture was stirred at room temperature over night. The mixture was filtered and dried under vacuum at 45° C. for 10 h to give the title compound as a white solid (4.0 g, 93%). The title compound was characterized by LC-MS and 1H NMR as shown below:
LC-MS (ESI, pos. ion) m/z: 612.1 [M−1];
1H NMR (400 MHz, D2O): δ 0.88 (t, J=7.4 Hz, 3H), 1.65-1.75 (m, 2H), 1.74 (d, J=5.9 Hz, 3H) 2.93 (t, J=7.5 Hz, 2H), 6.55-6.62 (m, 1H), 7.04 (t, J=8.8 Hz, 1H), 7.40 (d, J=8.4 Hz, 2H), 7.37-7.44 (m, 1H), 7.55 (d, J=8.2 Hz, 2H), 8.32 (br, 1H), 8.41-8.47 (m, 1H), 8.55 (d, J=1.8 Hz, 1H).
BIOLOGICAL TESTING
The efficacy of the compounds disclosed herein can be evaluated as follows.
Example A
General Bio-Analysis Method
The LC/MS/MS system used in the analysis consists of an Agilent 1200 Series vacuum degasser, binary pump, well-plate autosampler, thermostated column compartment, the Agilent G6430 TripleQuadrupole Mass Spectrometer with an electrospray ionization (ESI) source. Quantitative analysis was carried out using MRM mode. The parameters for MRM transitions are in the Table A.









TABLE A








MRM
490.2→383.1



Fragmentor
 230 V



CE
  55 V



Drying Gas Temp
350° C.



Nebulize
  40 psi



Drying Gas Flow
  10 L/min









An Agilent XDB-C18, 2.1×30 mm, 3.5 μM column was used for the analysis. 5 μL of the samples were injected. Analysis condition: The mobile phase was 0.1% formic acid in water (A) and 0.1% formic acid in methanol (B). The flow rate was 0.4 mL/min. And the gradient of Mobile phase was in the Table B.









TABLE B






Time
Gradient of Mobile Phase B








0.5 min
5%



1.0 min
95%



2.2 min
95%



2.3 min
5%



5.0 min
stop









Alternatively, an Agilent 6330 series LC/MS/MS spectrometer equipped with G1312A binary pumps, a G1367A autosampler and a G1314C UV detector was used in the analysis. An ESI source was used on the LC/MS/MS spectrometer. The analysis was done in positive ion mode as appropriate and the MRM transition for each analyte was optimized using standard solution. A Capcell MP-C18 100×4.6 mm I.D., 5 nM column (Phenomenex, Torrance, Calif., USA) was used during the analysis. The mobile phase was 5 mM ammonia acetate, 0.1% MeOH in water (A): 5 mM ammonia acetate, 0.1% MeOH in acetonitrile (B) (70:30, v/v). The flow rate was 0.6 mL/min. Column was maintained at ambient temperature. 20 μL of the samples were injected.
Example B
Compound Stability in Human and Rat Liver Microsomes
Human or rat liver microsomes incubations were conducted in duplicate in polypropylene tubes. The typical incubation mixtures consisted of human (or rat) liver microsomes (0.5 mg protein/mL), compounds of interest (5 μM) and NADPH (1.0 mM) in a total volume of 200 μL potassium phosphate buffer (PBS, 100 mM, pH7.4). Compounds were dissolved in DMSO and diluted with PBS such that the final concentration of DMSO was 0.05%. The enzymatic reactions were commenced with the addition of protein after a 3 min preincubation and incubated in a water bath open to the air at 37° C. Reactions were terminated at various time points (0, 5, 10, 15, 30, 60 min) by adding equal volume of ice-cold acetonitrile. The samples were stored at −80° C. until LC/MS/MS assays.
The concentrations of compounds in the incubation mixtures of human (or rat) liver microsomes were determined by a LC/MS/MS method. The ranges of the linearity in the concentration range were determined for each tested compounds.
A parallel incubation was performed using denatured microsomes as the negative control, and reactions were terminated at various time points (for example, 0, 15, 60 min) after incubation at 37° C.
Dextromethorphan (70 μM) was selected as the positive control, and reactions were terminated at various time points (0, 5, 10, 15, 30, 60 min) after incubation at 37° C. Both positive and negative control samples were included in each assay to ensure the integrity of the microsomal incubation system.
Alternatively, the stability of some of the compounds disclosed herein in human (or rat) liver microsomes were also conducted in the following protocol. The incubations were conducted in duplicate in polypropylene tubes. The typical incubation mixtures consisted of liver microsomes (final concentration: 0.5 mg protein/mL), compounds (final concentration: 1.5 μM) in a total volume of 30 μL K-buffer (contain 1.0 mM EDTA, 100 mM, pH7.4). Compounds were dissolved in DMSO and diluted with K-buffer such that the final concentration of DMSO was 0.2%. The enzymatic reactions were commenced with the addition of 15 μL of NADPH (final concentration: 2 mM) after 10 min preincubation and incubated in a 37° C. incubator. Reactions were terminated at various time points (0, 15, 30, 60 min) by adding 135 μL acetonitrile (contain IS). Protein is removed by centrifugation with 4,000 rpm, 10 min.
Supernatant was Collected for LCMS/MS Analysis
In the above protocol, ketanserin (1 μM) was selected as the positive control, and reactions were terminated at various time points (0, 15, 30, 60 min) after incubation at 37° C. The positive control sample was included in each assay to ensure the integrity of the microsomal incubation system.
Data Analysis
The concentrations of compounds in liver microsome incubations were plotted as a percentage of the relevant zero time point control for each reaction. The in vivo CLint were extrapolated (ref: Naritomi et al., Prediction of human hepatic clearance from in vivo animal experiments and in vitro metabolic studies with liver microsomes from animals and humans, Drug Metab. Dispos., 2001, 29: 1316-1324.)
Exemplary results from selected compounds of the invention are listed in Table 2. The compounds disclosed herein exhibited desirable half-life (T1/2) and clearance (CLint) when the compounds were incubated in human and rat liver microsomes. Similarly, selected compounds of the invention were also assessed for their stability in human and rat plasma, and the results are listed in Table 3.







TABLE 2







Stability of selected compounds of the invention in human and rat liver


microsomes (1 μM)












Human

Rat















T1/2
CLint
T1/2
CLint



Example #
(min)
(mL/min/kg)
(min)
(mL/min/kg)
















Ex. 1
28.89
60.17
17.54
141.60



Ex. 2
59.49
29.22
12.85
193.28



Ex. 7a
∞
ND
53.06
46.81













Ex. 9
FTV

NT
NT













Ex. 10
FTV

FTV




Ex. 11
FTV

FTV














Ex. 14
33.62
51.7
NT
NT



Ex. 18
78.77
5.52
NT
NT













Ex. 19
FTV

FTV




Ex. 20
FTV

FTV




Ex. 21
FTV

FTV














Ex. 22
72.8
23.9
4.6
535.4



Ex. 23
112.8
15.4
5.8
429.2



Ex. 25
∞
ND
∞
ND













Ex. 26
FTV

NT
NT













Ex. 27
FTV

FTV




Ex. 28
FTV

FTV














Ex. 32
∞
ND
∞
ND





FTV: Fast Transformed to Vemurafenib;


NT: not tested;


ND: not determined













TABLE 3







Stability of selected compounds of the invention in human


and rat plasma (2 μM)












Human
Rat




T1/2
T1/2



Example #
(min)
(min)














Ex. 7a
∞
68.82



Ex. 9
112.9
28.55



Ex. 10
88.12
55.78



Ex. 11
162.2
49.80



Ex. 14
510.8
NT



Ex. 15
39.61
18.57



Ex. 16
∞
157.3



Ex. 18
6.08
9.61



Ex. 19
7.47
5.90



Ex. 22
6.81
15.24



Ex. 23
3.06
3.42



Ex. 26
81.6
NT



Ex. 27
79.63
55.85



Ex. 28
57.87
49.55



Ex. 30
∞
56.32





NT: not tested






Example C
Evaluation of Pharmacokinetics after Intravenous and Oral Administration of the Compounds Disclosed Herein in Mice, Rats, Dogs and Monkeys
Selected compounds of the invention and Vemurafenib were assessed in pharmacokinetic studies in mice, rats, dogs or monkeys. The compounds were administered as a water solution, 2% HPMC+1% TWEEN® 80 in water solution, 5% DMSO+5% solutol in saline, 4% MC or capsule. For the intravenous administration, the animals were generally given at 1 or 2 mg/kg dose. For the oral (p.o.) dosing, mice and rats were generally given 5 or 10 mg/kg dose, and dogs and monkeys were generally given 7-100 mg/kg dose. The blood samples (0.3 mL) were drawn at 0.25, 0.5, 1.0, 2.0, 3.0, 4.0, 6.0, 8.0, 12 and 24 h time points or 0.083, 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 and 24 h time points and centrifuged at 3,000 or 4000 rpm for 2 to 10 min. The plasma solutions are collected and stored at −20° C. or −70° C. before analyzed by LC/MS/MS as described above.
Exemplary study results from examples disclosed herein are listed in Table 4 to Table 8. The compounds disclosed herein exhibited optimized pharmacokinetic properties with desirable bioavailability (F) and half-life (T1/2) when the compounds were administered intravenously or orally.







TABLE 4







Vemurafenib's Pharmacokinetic profiles in SD rats














Dose


Cmax
AUClast



Sample
mg/kg
T1/2 h
Tmax h
ng/ml
ng · h/mL
F %
















Vemurafenib
PO 10
3.54
4
25420
261424
104



IV 1
3.75
NT
NT
25217
/





NT: not tested













TABLE 5







Pharmacokinetic profiles of selected compounds of the


invention in SD rats (PO, 10 mg/kg dosing)















Cmax
AUClast
F


Example #
T1/2 h
Tmax h
ng/ml
ng · h/mL
(Vemurafenib) %















Ex. 1
3.7
2.67
21317
171658
78.1


Ex. 2
5.8
3.33
14801
134580
64.7


Ex. 7a
3.82
2.33
28468
272090
133


Ex. 9
2.57
1.67
25875
194649
93


Ex. 10
2.39
1.333
25889
232744
111


Ex. 11
3.77
2
23022
226462
113


Ex. 14
2.97
2.667
27645
196280
96.0


Ex. 15
2.63
1.67
31024
273056
127


Ex. 16
4.91
3.33
5806
76466
39.1


Ex. 18
3.47
4.67
17473
229429
115


Ex. 19
7.78
4.00
20050
311059
145


Ex. 20
2.50
2.00
25460
169136
80.9


Ex. 21
2.69
1.33
19643
118729
58.2


Ex. 22
3.10
1.67
18935
182035
187


Ex. 23
3.46
1.33
23106
183843
188


Ex. 25
3.25
2.67
12774
114018
59.4


Ex. 26
2.36
1.67
29520
170624
81.8


Ex. 27
3.56
3.33
62049
348111
171


Ex. 28
3.28
1.67
15870
154036
75.6


Ex. 30
3.84
2.67
24882
252715
127
















TABLE 6







Vemurafenib's Pharmacokinetic profiles in beagle dogs














Dosing


Cmax
AUClast



Sample
mg/kg
T1/2 h
Tmax h
ng/ml
ng · h/mL
F %
















Vemurafenib
PO 10
3.9
2
2149
16537
33.4



IV 1
1.59
NT
NT
4877
/





NT: not tested













TABLE 7







Pharmacokinetic profiles of selected compounds of


the invention in beagle dogs (PO)














PO




F



Dosing

Tmax
Cmax
AUClast
(Vemurafenib)


Example #
mg/kg
T1/2 h
h
ng/ml
ng · h/mL
%
















Ex. 25
10
1.94
4
2208
19057
47.9



40
1.95
4
12843
144162
90.5



70
1.84
2
38
173008
62.1


Ex. 32
30
1.37
4
22200
125254
109



50
1.71
4
29900
210190
183
















TABLE 8







Pharmacokinetic profiles of Example 25 in Monkeys (PO)













PO







Dosing


Cmax
AUClast


Example #
mg/kg
T1/2 h
Tmax h
ng/ml
ng · h/mL















Ex. 25
50
9.32
4
22136
225569



50
12.07
4
23038
247859



100
27.04
4
34793
467762









Example D
Tumor Xenograft Models
The efficacy of compounds disclosed herein is evaluated in a standard murine model of tumorigenesis. Human tumor cells (i.e., Colo-205 Tumor Xenograft models) are expended in culture, harvested, and injected subcutaneously onto the rear flank of 6-7 week old female athymic nude mice (BALB/cA nu/nu, Shanghai SLAC Laboratory Animal, Co.) (n=10 for vehicle group, n=8 for each dosing group). When tumor reaches a volume of 100-250 mm3, animals are randomly divided into vehicle control (for example, 5% PEG400+5% Solutol HS 15 in water or 5% EtOH+5% CremophorEL+1% HCl (11.7%), pH 2.2 in water) and compound groups. Subsequent administration of compound by oral gavage (for example, 6-60 mpk/dose, dissolved in 5% PEG400+5% Solutol HS 15 in water or 5% EtOH+5% CremophorEL+1% HCl (11.7%), pH 2.2 in water) begins anywhere from day 0 to day 15 post tumor cell challenge and generally continues with once a day for the duration of the experiment.
Tumor Growth Inhibition (TGI) Analysis
Progression of tumor growth is assessed by tumor volumes and recorded as a function of time. The long (L) and short (W) axes of the subcutaneous tumors are measured with calipers twice weekly, and the tumor volume (TV) calculated as (L×W2)/2). TGI is calculated from the difference between the median tumor volumes of vehicle-treated and drug-treated mice, expressed as a percentage of the median tumor volume of the vehicle-treated control group, by the following relation:




  
    %
    ⁢
    
      
    
    ⁢
    TGI
  
  =
  
    
      (
      
        
          
            
              
                
                  Median
                  ⁢
                  
                    
                  
                  ⁢
                  Tumor
                  ⁢
                  
                    
                  
                  ⁢
                  
                    Volume
                    control
                  
                
                -
              
            
          
          
            
              
                Median
                ⁢
                
                  
                
                ⁢
                Tumor
                ⁢
                
                  
                
                ⁢
                
                  Volume
                  
                    drug
                    ⁢
                    
                      -
                    
                    ⁢
                    
                      
                    
                    ⁢
                    treated
                  
                
              
            
          
        
        
          Median
          ⁢
          
            
          
          ⁢
          Tumor
          ⁢
          
            
          
          ⁢
          
            Volume
            control
          
        
      
      )
    
    ×
    100
  




Initial statistical analysis is done by repeated measures analysis of variance (RMANOVA), Followed by Scheffe post-hoc testing for multiple comparisons. Vehicle alone (2% HPMC+1% Tween-80, or the like) is the negative control.
The compouds described herein were also administrated orally (p.o.) once a day (QD) or (BID) for 15 days in Colo-205 xenograft animal model. At doses of 20 mg/kg, the compouds produced statistically significant inhibition of growth of certain tumors grown subcutaneously in athymic nude mice. Exemplary xenograft study results from Examples 25, 32 are listed in Table 9.







TABLE 9







TGI % of Colo-205 Xenograft models








TGI %
Colo-205 Xenograft models











(on the last day of

20
20
60


dosing)
6 mg/kg/qd
mg/kg/qd
mg/kg/bid
mg/kg/qd














Vemurafenib (15 days)
33
73
101
104


Ex. 25 (15 days)
41
59
95
108


Ex. 32 (15 days)
29
68
102
110









The compounds disclosed herein exhibited optimized pharmacokinetic properties with desirable clearance (CL), half-life (T1/2) and excellent bioavailability when the compounds were administered intravenously or orally.
Finally, it should be noted that there are alternative ways of implementing the present invention. Accordingly, the present embodiments are to be considered as illustrative and not restrictive and the invention is not be limited to the details given herein, but may be modified within the scope and equivalents of the appended claims. All publications and patents cited herein are incorporated by reference.

## Claims

1. A compound of Formula (I):



or a racemic mixture, a stereoisomer, a geometric isomer, a tautomer, a solvate, an N-oxide, a metabolite or a pharmaceutically acceptable salt thereof, wherein:
each of X and Y is independently H, —C(═O)R3, —C(═O)OR4, —C(R1R2)OC(═O)R3, —C(R1R2)OC(═O)OR4 or —C(R1R2)OP(═O)(OR4)(OR4a), with the proviso that when Y is H, X is not —C(═O)CH3 or H;

each R1 and R2 is independently H, D, C1-6alkyl, C1-6haloalkyl, C3-6cycloalkyl, —(C1-4 alkylene)-(C3-6cycloalkyl), C3-6heterocyclyl or —(C1-4alkylene)-(C3-6heterocyclyl); or R1 and R2, together with the carbon atom they are attached to, form an unsubstituted or substituted 3-8 membered carbocyclic or heterocyclic ring;
each R3 is independently H, D, C1-10haloalkyl, C3-8cycloalkyl, —(C1-6 alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl or —(C1-6alkylene)-(5-10 membered heteroaryl), wherein each of the C1-10alkyl, C1-10haloalkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6 alkylene)-(C6-10aryl), 5-10 membered heteroaryl and —(C1-6alkylene)-(5-10 membered heteroaryl) is unsubstituted or optionally substituted with 1, 2, 3 or 4 substituents independently selected from D, F, Br, I, C1-6alkyl, C1-6haloalkyl, OH, NH2, C1-6 alkoxy and C1-6alkylamino; and
each R4 and R4a is independently H, C1-10haloalkyl, C3-8cycloalkyl, —(C1-6 alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl or —(C1-6alkylene)-(5-10 membered heteroaryl), wherein each of the C1-10alkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10aryl, —(C1-6 alkylene)-(C6-10aryl), 5-10 membered heteroaryl and —(C1-6alkylene)-(5-10 membered heteroaryl) is unsubstituted or optionally substituted with 1, 2, 3 or 4 substituents independently selected from D, F, Cl, OH, NH2, oxo (═O), C1-6haloalkyl, C1-6 alkoxy and C1-6alkylamino.

2. The compound of claim 1, wherein each R1 and R2 is independently H, D or C1-3 alkyl.

3. The compound of claim 1, wherein each R3 is independently C1-10alkyl, C3-8 cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8 heterocyclyl), C6-10aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl or —(C1-6 alkylene)-(5-10 membered heteroaryl), wherein each of the C1-10alkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl), C6-10 aryl, —(C1-6alkylene)-(C6-10aryl), 5-10 membered heteroaryl and —(C1-6alkylene)-(5-10 membered heteroaryl) is unsubstituted or optionally substituted with 1, 2, 3 or 4 substituents independently selected from D, F, OH, OMe, NH2, NHMe, NMe2 and C1-3 alkyl.

4. The compound of claim 1, wherein each R4 and R4a is independently H, C1-10 alkyl, C3-8cycloalkyl, —(C1-6alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl) or C6-10aryl, wherein each of the C1-10alkyl, C3-8cycloalkyl, —(C1-6 alkylene)-(C3-8cycloalkyl), C3-8heterocyclyl, —(C1-6alkylene)-(C3-8heterocyclyl) and C6-10 aryl is unsubstituted or optionally substituted with 1, 2, 3 or 4 substituents independently selected from D, F, OH, OMe, oxo (═O), NH2, NHMe, NMe2 and C1-3 alkyl.

5. The compound of claim 1, wherein each of X and Y is independently H or —C(R1R2)OP(═O)(OH)2, with the proviso that when Y is H, X is not H.

6. The compound of claim 1, wherein each of X and Y is independently H, —C(═O)R3, —C(═O)OR4, —C(R1R2)OC(═O)R3 or —C(R1R2)OC(═O)OR4, with the proviso that when Y is H, X is not —C(═O)CH3 or H.

7. The compound of claim 6, wherein the acyl moiety (—C(═O)R3) is derived from an α-amino acid or an optical isomer thereof by removing the OH group from the carboxy group of the α-amino acid.

8. The compound of claim 7, wherein the α-amino acid is isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, alanine, asparagine, aspartate, glutamate, glutamine, proline, serine, para-tyrosine, arginine, histidine, cysteine, glycine, sarcosine, N,N-dimethyl glycine, homoserine, norvaline, norleucine, ornithine, homocysteine, homophenylalanine, phenylglycine, ortho-tyrosine, meta-tyrosine or hydroxyproline.

9. The compound of claim 8, wherein the α-amino acid is isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, alanine, asparagine, aspartate, glutamate, glutamine, proline, serine, tyrosine, arginine or histidine, each of which has a S-configuration at the α-position.

10. The compound of claim 1, wherein the pharmaceutically acceptable salt is an alkali metal salt, an alkaline earth metal salt, or an ammonium or N+(C1-4 alkyl)4 salt.

11. The compound of claim 10, wherein the pharmaceutically acceptable salt is a sodium salt, a lithium salt, a potassium salt, a calcium salt, a magnesium salt, an ammonium salt, a quaternary ammonium salt, or a combination thereof.

12. The compound of claim 1, wherein the pharmaceutically acceptable salt is a salt of an inorganic acid, an organic acid or a combination thereof, wherein the inorganic acid or the organic acid is hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, malic acid, 2-hydroxypropanic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, glucuronic acid, galacturonic acid, citric acid, tartaric acid, aspartic acid, glutamic acid, benzoic acid, cinnamic acid, p-toluenesulfonic acid, benzenesulfonic acid, mthanesulfonic acid, ethanesulfonic acid, trifluoromthanesulfonic acid or a combination thereof.

13. The compound of claim 1 having one of the following structures:

14. A pharmaceutical composition comprising the compound of claim 1, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle or a combination thereof.