EP4583870A2 - Mitochondrial uncouplers for treatment of metabolic diseases and cancer - Google Patents

Mitochondrial uncouplers for treatment of metabolic diseases and cancer

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Publication number
EP4583870A2
EP4583870A2 EP23863911.6A EP23863911A EP4583870A2 EP 4583870 A2 EP4583870 A2 EP 4583870A2 EP 23863911 A EP23863911 A EP 23863911A EP 4583870 A2 EP4583870 A2 EP 4583870A2
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EP
European Patent Office
Prior art keywords
group
compound
alkyl
rule
substitute sheet
Prior art date
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EP23863911.6A
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German (de)
French (fr)
Inventor
Shengkan Jin
David J. Augeri
Bin Cao
Hanlin TAO
Nigel Liverton
Casey Mccomas
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Mito Biopharma LLC
Rutgers State University of New Jersey
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Mito Biopharma LLC
Rutgers State University of New Jersey
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Publication of EP4583870A2 publication Critical patent/EP4583870A2/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D277/00—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings
    • C07D277/60—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings condensed with carbocyclic rings or ring systems
    • C07D277/62—Benzothiazoles
    • C07D277/68—Benzothiazoles with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached in position 2
    • C07D277/82—Nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
    • C07H15/26—Acyclic or carbocyclic radicals, substituted by hetero rings

Definitions

  • each of substituents R 5000A andR 5000B is independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH 2 ) 2 OCH3; alternatively R 5000A and R 5000B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl;
  • Z 1000 is selected from the group consisting of H, -CH 2 OCH3, -CH 2 OCH 2 CH3, -CH 2 O(CH 2 ) 2 OH, -CH 2 O(CH 2 ) 2 OCH 3 , -CH 2 O(CH 2 ) 2 N(CH 3 ) 2 , -CH 2 O(CH 2 ) 2 NHSO 2 CH 3 , -(CH2)O(CH 2 ) 2 NR 2000A R 2000B , -(CH 2 ) S R 3000 , -CI FOCI F Ar 1 , OCH3 CH 2 NHC(O)CH 2 CH 3 , -CH 2 NHC(O)CH 2 OCH 3 , -CH 2 NHSO 2 CH 3 , -(CH 2 )t’NR 7000A R 7000B , and -(CH 2 ) t R 8000 ;
  • R 3000 is a 5 to 6-membered heterocyclic ring
  • Ar 1 is a 5 to 6-membered aryl or heteroaryl group optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl, halo, hydroxyl and alkoxy;
  • each of R 7000A and R 7000B is independently selected from Ci-Ce alkyl; alternatively, R7OOOA an j R 7000B t O g e th er with the nitrogen to which they are attached, form a 4 to 8- membered heterocyclyl optionally substituted with one or more substituents independently selected from Ci-Ce alkyl;
  • R 5000A and R 5000B are not both Ci-Ce alkyl; and when Z 1000 is H; R 1000a is not Ci- C 6 alkyl, -C 3 -C 6 cycloalkyl, CH 3 or CH 2 CH 3; or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
  • each of R 5A andR 5B is independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH 2 ) 2 OCH 3 ; alternatively R 5A and R 5B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino, carboxyamido, -SO 2 CH 3 , -CF 3 , Ci-Ce alkyl, halo, and acyl;
  • R 6 is selected from the group consisting of a 5 to 6-membered heterocyclyl, pyridinyl and thiazolyl; m’ is an integer selected the group consisting of 1, 2 and 3; m is an integer selected the group consisting of 0, 1, 2 and 3;
  • R lc is selected from the group consisting of chloro, fluoro, iodo, and bromo; each of R 4b and R 4d is independently selected from the group consisting of Y and Z, provided that when R 4b is Y, R 4d is Z and when R 4b is Z, R 4d is Y;
  • Y is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF 3 , -CHF 2 , fluoro(Ci-C6)alkyl, halo(Ci-C6)alkyl, -OCF 3 , -SO 2 (Ci-C6)alkyl, cyano, and -CO 2 (Ci-C 6 )alkyl;
  • R B is a conventional mitochondrial uncoupler prior to being covalently linked to R A ; provided the mitochondrial membrane-retaining uncoupler compound is not
  • Oligo is oligomycin 2.5 pM; AA isantimycin A 2 pM; Rot isrotenone 2 pM.
  • Figure 4B shows mitochondrial membrane potential (MMP) determined by fluorescence microscopy using two different dyes, tetramethylrhodamine ethyl ester (TMRE) staining and DilCl(5) staining, respectively, in the presence of FCCP at the indicated concentrations. The fluorescent intensity is indicative of MMP.
  • FCCP has a ratio of CIO%TMRE / Cmin-OCR less than 3, where CIO%TMRE is the concentration leading to 10% MMP retention (or 90% MMP loss, measured by TMRE staining) and Cmin-ocR is the minimal concentration leading to OCR increase.
  • SUBSTITUTE SHEET (RULE 26 ) represents Compound 25 at 9.0 pM, and the circle represents Compound 25 atl2.0 pM.
  • Oligo isoligomycin 2.5 pM; AA is antimycin A 2 pM; Rot isrotenone 2 pM.
  • Figure 5B shows mitochondrial membrane potential (MMP) determined by fluorescence microscopy using two different dyes, tetramethylrhodamine ethyl ester (TMRE) staining and DilCl(5) staining, respectively, in the presence of Compound 25(#25) at the indicated concentrations. The fluorescent intensity is indicative of MMP.
  • Compound 25 represents the MMP -retaining uncouplers that do not cause observable MMP reduction while uncoupling mitochondria (with a ratio of CIO%TMRE I Cmin-ocR greater than 25).
  • Figure 6 shows the effect of Compound 64 (#64), on oxygen consumption rate (Figure 6A) and mitochondrial membrane potential (Figure 6B).
  • Compound 64 increases OCR without drastically dissipating mitochondrial membrane potential.
  • Figure 6A shows cellular oxygen consumption rates determined by Seahorse OCR assay using C2C12 cells at the indicated concentrations.
  • the diamond in Figure 6A represents vehicle treatment control, the square represents Compound 64 (#64) at 0.3 pM, the triangle represents Compound 64 at 1.0 pM, the X represents Compound 64 at 2.0 pM, the * represents Compound 64 at 3.0 pM, and the circle represents Compound 64 at 4.0 pM.
  • Oligo is oligomycin 2.5 pM; AA is antimycin A 2 pM; Rot is rotenone 2 pM.
  • Figure 6B shows mitochondrial membrane potential (MMP) determined by fluorescence microscopy using two different dyes, tetramethylrhodamine ethyl ester (TMRE) staining and DilCl(5) staining, respectively, in the presence of Compound 64 (Comp #64) at the indicated concentrations. The fluorescent intensity is indicative of MMP.
  • Compound 64 represents the MMP-retaining uncouplers that do not drastically reduce MMP while uncoupling mitochondria (with a ratio of CIO%TMRE / Cmin-ocR between 10 and 25).
  • Figure 7 shows a schematic illustration of the mechanism of action of a conventional mitochondrial uncoupler ( Figure 7A) and a proposed mechanism of action of an MMP-retaining uncoupler (Figure 7B), as well as diagrams and an example of how to convert a conventional uncoupler to an MMP-retaining uncoupler ( Figure 7C-E).
  • Figure 7A and Figure 7B show mitochondrial electron transport chain complexes I, II, III, IV, ATP synthase, as well as uncouplers (UH or U").
  • Conventional uncouplers allow proton translocation across the mitochondrial inner membrane, causing loss of MMP (Figure 7A).
  • SUBSTITUTE SHEET ( RULE 26 ) conventional uncoupler ( Figure 7C), for example, by adding a tertiary amine- (or secondary amine-) containing side chain to obtain an MMP -retaining uncoupler (Figure 7D), where N is nitrogen; and X and Y are optimally substituted side chains.
  • Figure 7E shows an example of an MMP -retaining uncoupler, showing a conventional uncoupling component in the frame.
  • Compounds 64 and 25 were prepared to a fine suspension in 0.5% CMC-Na / 1% Tween80 water solution. The tests were performed under fed condition, by oral gavage of 100-400 pL compound suspension according to the body weight to desired dosages (mg/kg). The mice were supplied with drinking water all through the testing time. Mouse behaviors were monitored every 15-30 minutes and LD50 were determined. MED was determined in diabetic and steatosis mouse models (Table 9).
  • Figures 9 shows the safety profile of Compound 64 (#64) compared to DNP.
  • Figure 9A shows the NOAEL(no-observable-adverse-effect-level)/MED of DNP. https://www.atsdr.cdc.gov/ToxProfiles/tp64.pdf; (US EP A, 2, 4-dinitrophenol. https://www.epa.gov/sites/default/files/2016-09/documents/2-4-dinitrophenol.pdf; and US CDC).
  • Figure 9B shows the MED, intermediate NOAEL and NOAEL/MED of Compound 64 calculated by oral dosage.
  • Figure 9C shows the MED, intermediate NOAEL and NOAEL/MED of Compound 64 over DNP calculated by Cmax (maximal blood concentration).
  • Figure 9D shows the MED, intermediate NOAEL and NOAEL/MED of Compound 64 calculated byAUC (Area Under Curve).
  • the Figures demonstrate that the MMP-retaining uncoupler, Compound 64 exhibits drastically improved short-term safety profiles over the conventional uncoupler DNP.
  • Figure 10 show the safety profile of Compound 25 (#25)
  • Figure 10A shows the MED, intermediate NOAEL and NOAEL/MED of Compound 25 calculated by oral dosage.
  • Figure 10B shows the MED, intermediate NOAEL and NOAEL/MED of Compound 25 over DNP calculated by Cmax (maximal blood concentration).
  • Figure 10C shows the MED, intermediate NOAEL and NOAEL/MED of Compound 25 calculated byAUC (Area Under Curve).
  • the Figures demonstrate that the MMP -retaining uncoupler, Compound 25 exhibits drastically improved short-term safety profiles over the conventional uncoupler DNP.
  • Figure 14 shows the effects of Compound 25 on blood triglyceride (Figure 14A), total cholesterol (Figure 14B), and nori-HDI. cholesterol levels (Figure 14C) in high-fat diet induced diabetic/hepatic steatosis mice.
  • Figure 18 shows the efficacy of Compound 25 in reducing liver fibrosis determined by histology and molecular analysis.
  • Figure 18A is microscope images of liver sections of CCL treated mice, subjected to H & E staining (top left panel); CCI4 plus Compound 25 (7.5 mg/kg/day) treated mice subjected to H & E staining (bottom left panel); CCL treated mice, subjected to Picrosirius Red staining(staining fibrotic collagen, top right panel) and CCL plus Compound 25 (7.5 mg/kg/day) treated mice subjected to Picrosirius Red staining (staining fibrotic collagen, bottom left panel.
  • Figure 19 is an immunoblotting analysis that shows the inhibitory effect of Compound 6464 on TGF-0 activation in T-cells.
  • Figure 19A is an immunoblotting analysis of Human Jurkat cells treated with either vehicle alone (first lanes), TGF-0 alone (second lanes), TGF-0 plus Compound 64 at 0.5, 1.0 or 2.0 pM of compound (third through 5th lanes), as indicated, for 6 hours.
  • Figure 19B is an immunoblotting analysis of mouse primary T-cells (B) were treated with either vehicle alone (first lanes), TGF-0 alone (second lanes), or TGF-0 plus varying concentrations of Compound 64 atl.O or 2.0 pM of compound (third and fourth lanes), as indicated, for 6 hours. Immunoblotting analyses were performed with antibodies against p-Smad2/3 (phosphorylated Smad2/3), Smad2/3, or GAPDH, as indicated.
  • Figure 20 shows Compound 64 is efficacious in combinatory therapy with PD-1 antibody in treating metastatic cancer in mice.
  • Figure 20A is the experimental design (see Example B 12). Briefly, C57/B16 mice intrahepatically transplanted with MC38 cancer cells (day 0) were subject to various treatments starting on day 7: aPD-1 or isotype, mice were either treated with PD-1 antibody or its isotype antibody (control) by intraperitoneal (IP) injection on indicated days (PD-1 antibody has a half-life of over 1 week in mice); #64 or vehicle, mice were either treated with Compound #64 or vehicle by daily gavage.
  • Figure 20B is a table depicting outcomes of the experiments described in Figure 20A and Example B12.
  • aPD-1 +#64 represents mice treated with PD-1 antibody (IP) and #64 (daily gavage); aPD-1 represents mice treated with PD-1 antibody (IP) and vehicle (gavage); control, represents mice treated with isotype antibody (IP) and vehicle (gavage); n is number of mice in each group. Tumor-positive is the number of tumor- bearing mice in each group; tumor- free is thenumber of tumor-free mice in each group; tumor-free % is thepercentage of tumor- free animals in each group. Fisher Exact 2x2 test is theP value comparing each experimental group to control group using statistical analysis with Fisher Exact 2x2 test. P ⁇ 0.05 is indicative of a statistically significant difference.
  • Figure 21 shows antiviral activity (EC50), cytotoxicity (TC50), and specificity index (SI) of Compounds 64, 25 and 57 against enveloped viruses.
  • EC50 isthe compound concentration that reduces virus- induced cytopathic effects (CPE) by 50%.
  • TC50 isthe compound concentration that leads to 50% of cell viability of uninfected cells; SI is the ratio between TC50 and EC50.
  • the experiments were performed as follows. The host cells, either Vero 760 or MRC-5, as indicated, were seeded in 96-well flat-bottom tissue culture plates and allowed to adhere overnight.
  • the cells were either infected with virus (either SARS-CoV-2 or alpha coronavirus 229E) or uninfected, and diluted test compounds were added to each well. Following incubation at 37°C, 5% CO2 for three days or six days, cell viability was determined. Percent of CPE reduction of the virus-infected wells and the percent of cell viability of uninfected drug control wells, were measured to calculate and determine the EC50 and TC50 values. SI, was calculated accordingly.
  • virus either SARS-CoV-2 or alpha coronavirus 229E
  • mitochondria are the ultimate site where lipid or glucose metabolites are consumed (oxidized); (2) mitochondria are critical in regulating the abundance of metabolic intermediates that become building blocks of biosynthesis essential for cell growth and proliferation of cancer cells, as well as for viral envelop production and assembly; (3) mitochondria are the major production site of ROS in neurons and many other cells ( Figure 1).
  • Mitochondrial uncoupling is a unique way to modulate mitochondrial activity and functions.
  • mitochondrial uncoupling is a process by which the activity of mitochondrial electron transport chain is de-coupled from ATP synthesis.
  • mitochondrial uncoupling is caused by the action of mitochondrial uncouplers that carry protons across the mitochondrial inner membrane into the mitochondrial matrix, independent of the ATP synthase (Terada, H. (1990) Environmental Health Perspectives 87, 213-218).
  • the technical definition of mitochondrial uncouplers is an increase of oxygen consumption rate (OCR) by cells in the presence of an ATP synthase inhibitor such as oligomycin.
  • OCR oxygen consumption rate
  • Metabolic diseases are a family of diseases characterized by symptoms of abnormal glucose and/or lipid metabolism, such as obesity, type 2 diabetes, alcoholic fatty liver disease, non-alcoholic fatty liver diseases, nonalcoholic steatohepatitis. These diseases are associated with age-, environmental-, or genetic- related decrease in mitochondrial functions such as reduced oxidative capacity. Importantly, these diseases also share a common causal factor, namely abnormal accumulation of intracellular lipid in cells of various tissues as well as insulin resistance in most cases. For example, obesity is characterized by excessive fat accumulation in cells of adipose tissue. Metabolic syndrome is characterized with insulin resistance in peripheral tissues, usually caused by ectopic fat accumulation in cells of liver, muscle, or adipose tissue. Type 2
  • Mitochondrial uncoupling reduces energy efficiency thereby undermining the energy requirement of cancer cells.
  • mitochondrial uncoupling promotes the complete mitochondrial oxidation of glucose and lipid, thereby diminishing the production of metabolic intermediates essential for biosynthesis of macromolecules required for cell proliferation.
  • mitochondrial uncoupling could lead to AMPK activation, a known event for inhibiting cell growth.
  • prior documents showed that mitochondrial uncouplers exhibit anti-cancer activities (U. S. Patent 10,227,315). Targeting cancer cells through mitochondrial uncoupling would deprive energy as well as biosynthetic metabolic intermediates that are absolutely essential for cancer cell growth and proliferation, which is proven to be an effective anti-cancer strategy (Alasadi, A. et al., (2016) Cell Death Dis., 9(2), 215)
  • Autoimmune diseases are conditions where the body’s immune system attacks their own healthy organs.
  • the common autoimmune diseases include celiac disease, diabetes mellitus type 1, Graves' disease, inflammatory bowel disease, multiple sclerosis, psoriasis, rheumatoid arthritis, and systemic lupus erythematosus.
  • the body’s own self-attaching immune cells need amplification (proliferation) which requires metabolic changes similar to the Warburg effect observed in cancer cells to provide sufficient building blocks for biosynthesis (Ganeshan, K., et al. (2014) Annual Review of Immunology, 32, 609-634). Therefore, mitochondrial uncoupling would potentially inhibit the activation and amplification of self-attacking immune cells.
  • Neurodegenerative diseases are a large group of disabling disorders of the nervous system such as Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and Alzheimer's disease, characterized by the relative selective death of neuronal subtypes.
  • Parkinson's disease Huntington's disease
  • amyotrophic lateral sclerosis amyotrophic lateral sclerosis
  • Alzheimer's disease characterized by the relative selective death of neuronal subtypes.
  • mitochondrial ROS production As one of the most important one (Elfawy, H. A., and Das, B. (2019) Life Sci., 218, 165-184.)
  • Mitochondrial uncouplers increase mitochondrial electron transport chain flux and decrease the electron stall in the complexes of the electron transport chain, thus they could effectively reduce mitochondrial ROS. Therefore, mitochondrial uncoupling is
  • Mitochondria are ancient bacteria that formed a symbiotic relationship with host cells. Bacterial plasma membrane contains electron transport chain and ATP synthase that are similar to those of mitochondria, therefore compounds that impact mitochondrial uncoupling could be useful inhibitors of bacterial growth and effective as antibiotics (US Patent 10,227,315).
  • Benzamide mitochondrial uncouplers have been developed (International Patent Publication Number WO 2012/068274, International Patent Publication Number WO 2016/081599, U. S. Patent 10,227,315, and Tao et al., 2014) for potential therapeutic applications.
  • One main limitation of the prior benzamide compounds are poor pharmacokinetic properties and low systemic exposure.
  • studies in animal models required the compound to be mixed with food, and high doses of the compounds were required to achieve efficacy (e.g. 1500 ppm niclosamide ethanolamine (NEN) in diet, equivalent to 150 mg/kg/day (Tao, et al.
  • MMP-retaining compounds exhibit drastically improved safety profiles compared to the bench mark conventional mitochondrial uncoupler, DNP. These compounds exhibit a wider therapeutic index when used for treating metabolic diseases.
  • 6-membered heterocyclyl is selected from the group consisting piperidinyl optionally substituted with one or Ci-Ce alkyl substituents, and morpholinyl optionally substituted with one or Ci-Ce alkyl substituents.
  • R la of Formula I, or of Formula la or a pharmaceutically acceptable salt, solvate, or prodrug thereof is -(CH2) m R 6 ; wherein R 6 is selected from the group consisting of tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, thiazolyl and piperidinyl and m is as previously described. In some embodiments m is 0. In some embodiments m is 1. In some embodiments -(CH2) m R 6 is selected from the group
  • R la of Formula I, or of Formula la or a pharmaceutically acceptable salt, solvate, or prodrug thereof is selected from the group consisting of -CH 3 , -CH2CH3, -OCH3, -CH2OCH3, -CH2OCH2OCH3, -CH 2 O(CH 2 )2OH, -CH 2 O(CH 2 )2OCH 3 , CH 2 )NH(CH 2 ) 2 OCH3, -CH 2 NHC(O)CH 3 , -(CH 2 )2NHCO 2 CH3,
  • R la of Formula I, or of Formula la or a pharmaceutically acceptable salt, solvate, or prodrug thereof is selected from the group consisting of -CH3, -CH2CH3 and -CH2NR 5A R 5B wherein R 5A and R 5B are as previously described in any embodiment described herein.
  • each of R 5A and R 5B is independently selected from Ci-Ce alkyl substituted with one or more methoxy; alternatively, R 5A and R 5B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl.
  • R lc of Formula I or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is chloro.
  • Some embodiments describe a compound of Formula I, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein R 4b is Y and R 4d is Z.
  • Y of Formula I, or a pharmaceutically acceptable salt, solvate, or prodrug thereof is -CF 3 .
  • Z of Formula I, or R 4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof is -(CH2)O(CH2)2NR 2A R 2B wherein R 2A and R 2B are as previously described herein.
  • R 2A and R 2B together with the nitrogen to which they are attached form a 6-membered heterocyclyl optionally substituted with a methyl group.
  • R 2A and R 2B together with the nitrogen to which they are attached form a heterocyclyl selected from piperazinyl or a 4- methyl piperazinyl.
  • Z of Formula I, or R 4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof is -(CH2) n R 3 , wherein R 3 is as previously described herein.
  • R 3 is a 5-membered heterocyclic ring.
  • R 3 is selected from the group consisting of tetrahydrofuranyl, and phenoxy.
  • R 3 is tetrahydrofuranyl.
  • R 3 is selected from the group consisting of and phenoxy.
  • n is 0.
  • n is 1.
  • -(CH2)nR 3 is selected from the group consisting
  • Z of Formula I, or R 4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof is -CH 2 OCH 2 Ar, wherein Ar is as previously described herein.
  • Ar is a 5 to 6-membered aryl or heteroaryl optionally substituted with one or more substituents independently selected from the group consisting of methyl, fluoro, chloro, hydroxy, and methoxy.
  • Ar is selected from the group consisting of thiazolyl, phenyl, and phenyl substituted with one or more groups independently selected from methyl, fluoro, chloro, hydroxy, and methoxy.
  • Ar is selected from the group consisting of, phenyl,
  • Z of Formula I, or R 4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof is selected from the group consisting of -CH 2 OH, -CH2OCH2CH3, -CH2OCH3, -CH 2 O(CH 2 )2OH, -CH 2 O(CH 2 )2OCH3, CH 2 O(CH2)2NHCH3,-CH 2 O(CH2)2N(CH3)2, -CH 2 O(CH 2 )2NHSO2CH3, -(CH 2 )O(CH 2 )2 NR 2A R 2B , -(CH2) n R 3 , -CH 2 OCH 2 Ar and OCH3; wherein R 2A R 2B , R 3 , n and Ar are as described in any embodiment herein.
  • Z of Formula I, or R 4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof is selected from the group consisting of -CH2OCH2CH -CH 2 O(CH 2 )2N(CH3)2, -CH 2 -CFLOCJTAr and OCH3; wherein R 2A R 2B , R 3 , n and Ar are as described in any embodiment herein.
  • Z of Formula I, or R 4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof is selected from the group consisting of -CH2OCH2CH3, -CH2OCH3, -CH 2 O(CH2)2OH, -CH 2 O(CH2) 2 OCH3, -CH 2 O(CH 2 ) 2 N(CH3)2, -CH 2 O(CH 2 )2NHSO2CH3, -(CH 2 )O(CH 2 ) 2 NR 2A R 2B , -(CH 2 ) n R 3 , -CFLOCJTAr and OCH3; wherein R 2A and R 2B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of piperazinyl and 4-methyl piperazinyl; R 3 is tetrahydrofuranyl; and Ar is a 5 to 6-membered aryl or heteroaryl group
  • SUBSTITUTE SHEET ( RULE 26 ) optionally substituted with one or more substituents independently selected from the group consisting of methyl, fluoro, chloro, hydroxy, and methoxy.
  • Z of Formula I, or R 4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof is selected from the group consisting of -CH 2 OH,-CH 2 OCH 2 CH 3 , -CH 2 OCH 3 , -CH 2 O(CH 2 )2OH, -CH 2 O(CH 2 )2OCH 3 ,
  • Z of Formula I, or R 4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group
  • Z of Formula I or R 4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof is selected from the group consisting of -CH 2 OH, -CH 2 OCH 2 CH 3 , -CH 2 OCH 3 , -CH 2 O(CH 2 ) 2 OH, -CH 2 O(CH 2 ) 2 OCH 3 ,
  • the compound is or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
  • the present disclosure describes a compound of
  • SUBSTITUTE SHEET ( RULE 26 ) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
  • R 50A and R 50B are independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH 2 ) 2 OCH 3 , with the proviso that R 50A and R 50B are not both Ci-Ce alkyl; alternatively R 50A and R 50B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido, -SO 2 CH 3 , -CF 3 , Ci-Ce alkyl
  • R 10a of Formula II, or of Formula Ila or a pharmaceutically acceptable salt, solvate, or prodrug thereof is selected from the group consisting of -CH2NR 50A R 50B ; wherein R 50A and R 50B together with the nitrogen to which they are attached, form a 4-7 membered heterocyclyl selected from the group consisting of azetindinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, morpholinyl, and dioxothiomorphylinyl; wherein the 4-7 membered heterocyclyl is optionally substituted with one or more substituents independently selected from the group consisting of oxo, cyano, hydroxyl, methoxy, -NHC(0)CH3, -C(0)NH2, -SO2CH3, -CF3, methyl, fluoro, and acetyl.
  • SUBSTITUTE SHEET ( RULE 26 ) substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl;
  • R 60 is selected from the group consisting of tetrahydrofuranyl, thiazolyl, and pyridinyl o is 0;
  • R 10c is selected from the group consisting of chloro, fluoro, iodo.
  • R 40d is H and R 40b is selected from the group consisting of fluoro, -CF3, -CHF2, and -OCF3;
  • R 10c is selected from the group consisting of chloro, fluoro, iodo.
  • R 10c is selected from the group consisting of chloro, fluoro, iodo.
  • R 40d is H and R 40b is selected from the group consisting of fluoro, -CF3, -CHF2, and -OCF3.
  • R 10a is selected from the group consisting of -OCH3, -CH2OCH3, -CH2OCH2OCH3, -CH 2 O(CH 2 )2OH, - CH 2 O(CH 2 ) 2 OCH3,
  • R 10c is chloro
  • R 40d is H and R 40b is -CF 3 .
  • R 10a is selected from the group consisting of -OCH 3 , -CH 2 OCH 3 , -CH 2 OCH 2 OCH 3 ,
  • Some embodiments describe a compound of Formula Ila or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein
  • SUBSTITUTE SHEET (RULE 26) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
  • R 100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof is selected from the group consisting of -CH2NR 500A R 500B ; wherein R 500A and R 500B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, -acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl.
  • the 4-8-membered heterocyclyl is selected from the group consisting of azetindinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, morpholinyl, dioxothiomorphylinyl, and azabicyclo[3.2.1]octanyl; wherein the heterocyclyl is optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, -acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl.
  • the 4-8-membered heterocyclyl is optionally substituted with one or more substituents independently selected from the group consisting of oxo, cyano, hydroxyl, methoxy, -NHC(O)CH3, -C(O)NH 2 , -SO2CH3, -CF3, methyl, fluoro, and acetyl. In some embodiments the 4-8-membered heterocyclyl is optionally substituted with one or more methyl substituents.
  • the 4 to 8-membered heterocyclyl is selected from the group consisting of azetindinyl; pyrrolidinyl; pyrrolidinyl substituted with one or more substituents selected from the group consisting of cyano, hydroxyl, methoxy, -NHC(O)CH3, - C(O)NH2, -SO2CH3, -CF3 and methyl; piperidinyl; piperidinyl substituted with one or more substituents selected from the group consisting of -CF3, and fluoro; piperazinyl substituted with one or more substituents selected from the group consisting of oxo, methyl, and acetyl; morpholinyl; morpholinyl substituted with one or more methyl groups; dioxothiomorphylinyl, and azabicyclo[3.2.1]octanyl.
  • R 100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof is selected from the group consisting of -CH2NR 500A R 500B ; wherein R 500A and R 500B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with one or more
  • SUBSTITUTE SHEET ( RULE 26 ) substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy,
  • 6-membered heterocyclyl is selected from the group consisting piperazinyl optionally substituted with one or Ci-Ce alkyl substituents, and morpholinyl optionally substituted with one or Ci-Ce alkyl substituents.
  • R 100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof is selected from the group consisting of -CH 2 NR 500A R 500B ; wherein -CH2NR 500A R 500B is selected from the group embodiments -CH2NR 500A R 500B is selected from the group consisting of
  • R 100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof is -(CH 2 ) P R 600 ; wherein R 600 is selected from the group consisting of tetrahydrofuranyl, tetrahydropyranyl, piperidinyl,
  • SUBSTITUTE SHEET ( RULE 26 ) thiazolyl and piperidinyl and m is as previously described.
  • p is 0.
  • p is 1.
  • -(CH 2 ) P R 600 is selected from the group
  • R 100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof is selected from the group consisting of -CH 3 , -OCH 3 , -CH 2 OCH 3 , -CH 2 OCH 2 OCH 3 , -CH 2 O(CH 2 )2OH, -
  • R 100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof is selected from the group consisting of -CH 3 , and -CH2NR 500A R 500B wherein R 500A and R 500B are as previously described in any embodiment described herein.
  • each of R 500A and R5°° B J S i n d e p encien tiy selected from Ci-Ce alkyl substituted with one or more methoxy; alternatively, R 500A and R 500B together with the nitrogen to which they are attached, form a 6- membered heterocyclyl optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl.
  • R 100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group
  • R 100c of Formula III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is chloro.
  • Some embodiments describe a compound of Formula III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein R 400b is Y 1 and R 400d is Z 1 .
  • Y'of Formula III or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is -CF 3 .
  • SUBSTITUTE SHEET ( RULE 26 ) embodiments R 7A and R 7B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of, 4-methylpiperazinyl, and morpholinyl; and R 8 is 4-methylpiperidinyl.
  • R 100a is selected from the group consisting of -CH3, -(CH2)N(CH2CH2OCH3)2,
  • R1°°C j s c hl oro - each of R 400b and R 400d is independently selected from the group consisting of Y 1 and Z 1 , provided that when R 400b is Y 1 , R 400d is Z 1 and when R 400b is Z 1 , R 400d is Y 1 ; Y 1 is CF3; and Z 1 is selected from the group consisting of -CH2NHC(O)CH2OCH3, -
  • R 100a is selected from the group consisting of -CH3, -(CH2)N(CH2CH2OCH3)2,
  • SUBSTITUTE SHEET (RULE 26) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
  • SUBSTITUTE SHEET (RULE 26) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
  • SUBSTITUTE SHEET (RULE 26 ) followed by liberation of the optically active bases from these salts.
  • An alternative process for separation of optical isomers includes the use of a chiral chromatography column optimally chosen to maximize the separation of the enantiomers.
  • Still another available method involves synthesis of covalent diastereoisomeric molecules by reacting compounds of the invention with an optically pure acid in an activated form or an optically pure isocyanate.
  • the synthesized diastereoisomers can be separated by conventional means such as chromatography, distillation, crystallization or sublimation, and then hydrolyzed to obtain the enantiomerically pure compound.
  • the optically active compounds of the invention can likewise be obtained by utilizing optically active starting materials. These isomers may be in the form of a free acid, a free base, an ester or a salt.
  • compositions according to embodiments described herein may be in the form of pharmaceutically acceptable salts.
  • a pharmaceutically acceptable salt of the compounds described herein includes acid addition salts and base addition salts.
  • Pharmaceutically-acceptable salt embraces salts commonly used to form alkali metal salts and to form addition salts of free acids or free bases The nature of the salt is not critical, provided that it is pharmaceutically-acceptable.
  • Suitable pharmaceutically-acceptable acid addition salts of the compounds described herein may be prepared from an inorganic acid or an organic acid. Examples of such inorganic acids include, without limitation, hydrochloric, hydrobromic, hydroiodic, nitric, carbonic, sulfuric and phosphoric acid.
  • the salt is a hydrochloride salt.
  • Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, arylaliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which include, without limitation, formic, acetic, propionic, succinic, glycolic, gluconic, maleic, embonic (pamoic), methanesulfonic, ethanesulfonic, 2- hydroxyethanesulfonic, pantothenic, benzenesulfonic, toluenesulfonic, sulfanilic, mesylic, cyclohexylaminosulfonic, stearic, algenic, P-hydroxybutyric, malonic, galactic, and galacturonic acid.
  • Salts derived from inorganic bases include by way of example only, sodium, potassium, lithium, ammonium, calcium and magnesium salts.
  • Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, such as alkyl amines, dialkyl amines, trialkyl amines, substituted alkyl amines, di(substituted alkyl) amines, tri(substituted alkyl) amines, alkenyl amines, dialkenyl amines, trialkenyl amines, substituted alkenyl amines, di (substituted alkenyl) amines, tri (substituted alkenyl) amines, cycloalkyl amines, di(cycloalkyl) amines, tri(cyclo alkyl) amines, substituted cycl
  • SUBSTITUTE SHEET (RULE 26 ) trisubstituted cycloalkyl amines, cycloalkenyl amines, di(cycloalkenyl) amines, tri(cycloalkenyl) amines, substituted cycloalkenyl amines, disubstituted cycloalkenyl amine, trisubstituted cycloalkenyl amines, aryl amines, diaryl amines, triaryl amines, heteroaryl amines, diheteroaryl amines, triheteroaryl amines, heterocyclic amines, diheterocyclic amines, triheterocyclic amines, mixed di- and tri-amines where at least two of the substituents on the amine are different and are selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalky
  • amines where the two or three substituents, together with the amino nitrogen, form a heterocyclic or heteroaryl group.
  • suitable amines include, by way of example only, isopropylamine, trimethyl amine, diethyl amine, tri (iso-propyl) amine, tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-alkylglucamines, theobromine, purines, piperazine, piperidine, morpholine, N-ethylpiperidine, and the like.
  • carboxylic acid derivatives would be useful in the preparation of pharmaceutically acceptable salts, for example, carboxylic acid amides, including carboxamides, lower alkyl carboxamides, dialkyl carboxamides, and the like.
  • Acceptable salts may be obtained using standard procedures well known in the art, for example by treating a sufficiently basic compound such as an amine with a suitable acid affording a physiologically acceptable anion.
  • a sufficiently basic compound such as an amine
  • a suitable acid affording a physiologically acceptable anion.
  • Alkali metal for example, sodium, potassium or lithium
  • alkaline earth metal for example calcium
  • prodrug forms Any compound that will be converted in vivo to provide the bioactive agent is a prodrug within the scope and spirit of the invention.
  • Various forms of prodrugs are well known in the art (see, for example, Medicinal Chemistry: Principles and Practice, F.D. King, ed., The Royal Society of Chemistry, Cambridge, UK, 1994; Hydrolysis in Drug and Prodrug Metabolism. Chemistry, Biochemistry and Enzymology, B. Testa, J. M. Mayer, VCHA and Wiley-VCH, Zurich, Switzerland, 2003; The Practice of Medicinal Chemistry, C. G. Wermuth, 2 nd ed., Academic Press, San Diego, CA, 1999).
  • Some prodrugs of the present invention include a compound according to any embodiment described herein in which the 2-
  • SUBSTITUTE SHEET ( RULE 26 ) hydroxy of the benzamide is converted to a group such as, but not limited to,
  • a prodrug of a compound according to any embodiment described herein may take the form of a carbamate.
  • the 2-hydroxy group of a benzamide according to any embodiment described may converted to a carbamate group, -OC(O)NR 9 R 10 at the same position.
  • R 9 and R 10 is independently selected from the group consisting of hydrogen, and optionally substituted Ci-Ce-alkyl; alternatively R 9 and R 10 taken together with the nitrogen to which they are attached form an optionally substituted Cs- -heterocyclyl.
  • the invention also embraces isolated compounds.
  • An isolated compound refers to a compound which represents at least 10%, preferably at least 20%, more preferably at least 50% and most preferably at least 80% of the compound present in the mixture.
  • one or more hydrogen atoms is replaced by a deuterium. It is well established that deuteration of physiologically active compounds offer the advantage of retaining the pharmacological profile of their hydrogen counterparts while positively impacting their metabolic outcome. Selective replacement of one or more hydrogen with deuterium, in a compound of the present invention, could improve the safety, tolerability and efficacy of the compound when compared to its all hydrogen counterpart.
  • Some embodiments describe a pharmaceutical composition
  • a pharmaceutical composition comprising: a compound according to an embodiment described herein, a pharmaceutically acceptable salt thereof, a solvate thereof, or a prodrug thereof; and a pharmaceutically acceptable carrier or diluent.
  • Compounds, or pharmaceutically acceptable salts thereof can be formulated for oral, intravenous, intramuscular, subcutaneous or parenteral administration for the therapeutic or prophylactic treatment of diseases, disorders or infections described herein.
  • SUBSTITUTE SHEET ( RULE 26 )
  • compounds of this invention can be mixed with conventional pharmaceutical carriers and excipients and used in the form of tablets, capsules, elixirs, suspensions, syrups, wafers and the like.
  • the pharmaceutical compositions comprising a compound of this invention will contain from about 0.1 to about 99% by weight of the active compound, and more generally from about 10 to about 30%.
  • compositions of the invention are prepared in accordance with standard procedures and are administered at dosages that are selected to reduce, prevent or eliminate the infection (See, e. g., Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa. and Goodman and Gilman's. The Pharmaceutical Basis of Therapeutics, Pergamon Press, New York, N.Y., the contents of which are incorporated herein by reference, for a general description of the methods for administering various agents for human therapy).
  • the pharmaceutical compositions of the invention can be delivered using controlled (e.g., capsules) or sustained release delivery systems (e.g., bioerodable matrices).
  • the pharmaceutically acceptable pharmaceutical compositions of the present invention comprise one or more compounds of the invention in association with one or more non-toxic, pharmaceutically acceptable carriers and/or diluents and/or adjuvants and/or excipients, collectively referred to herein as “carrier” materials, and if desired other active ingredients.
  • the pharmaceutical compositions may contain common carriers and excipients, such as com starch or gelatin, lactose, sucrose, microcrystalline cellulose, kaolin, mannitol, dicalcium phosphate, sodium chloride and alginic acid.
  • the pharmaceutical compositions may contain croscarmellose sodium, microcrystalline cellulose, com starch, sodium starch glycolate and alginic acid.
  • Tablet binders that can be included are acacia, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone (Povidone), hydroxypropyl methylcellulose, sucrose, starch and ethylcellulose.
  • Lubricants that can be used include magnesium stearate or other metallic stearates, stearic acid, silicone fluid, talc, waxes, oils and colloidal silica.
  • Flavoring agents such as peppermint, oil of wintergreen, cherry flavoring or the like can also be used. It may also be desirable to add a coloring agent to make the dosage form more aesthetic in appearance or to help identify the product.
  • the pharmaceutical compositions are in the form of, for example, a tablet, capsule, suspension or liquid.
  • the pharmaceutical composition is preferably made in the form of a dosage unit containing a therapeutically effective amount of the active ingredient. Examples of such dosage units are tablets and capsules.
  • the tablets and capsules which can contain, in addition to the active ingredient, conventional carriers such as binding agents, for example, acacia gum, gelatin, polyvinylpyrrolidone, sorbitol, or tragacanth; fillers, for example, calcium phosphate, glycine, lactose, maize-starch, sorbitol, or sucrose; lubricants, for example, magnesium stearate, polyethylene glycol, silica, or talc, disintegrants, for example, potato starch, flavoring or coloring agents, or acceptable wetting agents.
  • binding agents for example, acacia gum, gelatin, polyvinylpyrrolidone, sorbitol, or tragacanth
  • fillers for example, calcium phosphate, glycine, lactose, maize-starch, sorbitol, or sucrose
  • lubricants for example, magnesium stearate, polyethylene glycol, silica, or talc, disintegrants,
  • Oral liquid preparations generally are in the form of aqueous or oily solutions, suspensions, emulsions, syrups or elixirs may contain conventional additives such as suspending agents, emulsifying agents, non-aqueous agents, preservatives, coloring agents and flavoring agents.
  • additives for liquid preparations include acacia, almond oil, ethyl alcohol, fractionated coconut oil, gelatin, glucose syrup, glycerin, hydrogenated edible fats, lecithin, methyl cellulose, methyl or propyl parahydroxybenzoate, propylene glycol, sorbitol, or sorbic acid.
  • IV intravenous
  • a compound according to the invention can be dissolved or suspended in any of the commonly used intravenous fluids and administered by infusion.
  • Intravenous fluids include, without limitation, physiological saline or Ringer's solution.
  • Intravenous administration may be accomplished by using, without limitation, syringe, minipump or intravenous line.
  • Formulations for parenteral administration can be in the form of aqueous or non-aqueous isotonic sterile injection solutions or suspensions. These solutions or suspensions can be prepared from sterile powders or granules having one or more of the carriers mentioned for use in the formulations for oral administration.
  • the compounds can be dissolved in polyethylene glycol, propylene glycol, ethanol, com oil, benzyl alcohol, sodium chloride, and/or various buffers.
  • the compounds of the present invention can be presented in liquid or semi-liquid form formulated in hydrophobic or hydrophilic bases as ointments, creams, lotions, paints or powders.
  • Compound 17 of the present invention unexpectedly has drastically decreased metabolic stability (from 2,132 minutes to 145 minutes, Example B7), and concomitant decreased oral half-life (from 66.9 hrs to 8.6 hrs, Example B8).
  • the structural modification does not decrease the mitochondrial uncoupling activity thus are expected to have better toxicology properties when used to treat chronic conditions or disorders, which require prolonged use .
  • the mitochondrial uncouplers described herein represent a fundamentally different category of mitochondrial uncouplers which effectively induce mitochondrial uncoupling (increase mitochondrial oxygen consumption in the presence of oligomycin) without significantly decreasing mitochondrial membrane potential over a wide concentration range ( Figures 5 and 6, Example B2).
  • Figure 5 shows that Compound 25 does not appear to reduce MMP over a wide concentration range where OCR increases and reaches maximal levels.
  • the ratio of CIO%TMRE / Cmin-ocR for Compound 25 is over 25.
  • Figure 6 shows that Compound 64 effectively induces mitochondrial uncoupling without significantly decreasing MMP over a wide concentration range.
  • the ratio of CIO%TMRE / Cmin-ocR for Compound 64 is between 10 to 25.
  • TMRE tetramethylrhodamine ethyl ester
  • Results are categorized into three groups, the Compound 25-like compounds are denoted as MMP -Retaining Uncoupling Compounds (CIO%TMRE / Cmin-OCR>25); The Compound 64-like compounds are denoted as MMP -Retaining Uncoupling Compounds (Cio%TMRE / Cmin-ocR between 10 to 25), and Conventional Uncouplers (CIO%TMRE / Cmin-ocR less or equal to 3). The results are summarized in Table 6.
  • CW%TMRE is the concentration leading to 10% MMP retention (or 90% MMP loss, measured by TMRE staining) and C m in-ocR is the minimal concentration leading to OCR increase
  • a MMP- retaining uncoupler molecule consists of two functional parts (Figure 7C).
  • the first part functions as a conventional uncoupler, which transports proton from the mitochondrial intermembrane space to the mitochondrial matrix ( Figure 7B).
  • the second part is a positively charged functional group that is poorly impermeable to the mitochondrial inner membrane.
  • This feature allows an asymmetrical distribution and orientation of the compounds across mitochondrial inner membrane, with higher concentration of the charged molecules in the intramembrane space than in the mitochondrial matrix, as well as with the positively charged moiety primarily distributed at the outer surface of the membrane (facing the intermembrane space, Figure 7B).
  • mitochondrial uncoupling occurs and consequently reduction of proton gradient across the membrane
  • the loss of membrane potential due to proton gradient reduction is compensated by the asymmetrical positive charge distribution across the membrane provided by MMP-retaining uncouplers.
  • the overall mitochondrial membrane potential is minimally impacted over a wide concentration range of the uncouplers.
  • FIG. 9-10 show that the ratio ofNOAELs (no-observable-adverse-effect-level) over MED of Compounds 64 is over 40 (between 40-57 using different parameters), NOAEL/MED ratio of Compound 25 is over 19, while NOAEL/MED ratio of DNP is reportedly less than 3.
  • Some embodiments describe a method of preparing a mitochondrial membrane-retaining mitochondrial uncoupler comprising
  • the moiety capable of becoming positively charge in a cellular environment is a secondary or tertiary amine moiety.
  • Some embodiments describe a method of preparing a mitochondrial membrane-retaining mitochondrial uncoupler comprising
  • the secondary or tertiary amino moiety or R A is selected from the group consisting of -(CH2)NH(CH2)2OCH3, -CH2NHC(O)CH3, - previously described in any embodiment described herein.
  • a method of treating a mitochondria-related condition or diorder, in a subject in need thereof comprises administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition according to embodiments described herein.
  • the mitochondria-related condition or disorder is a metabolic disease.
  • the metabolic disease is selected from the group consisting of type 2 diabetes, a disease characterized by insulin resistance or hyperglycemia; obesity or obesity related complications, and a disease characterized by abnormal lipid accumulation.
  • a metabolic disease or disorder described in any embodiment herein is a complication caused by type 2 diabetes, selected from the group consisting of diabetes-induced cardiovascular diseases, neurodegenerative disorders, atherosclerosis, hypertension, coronary heart diseases, nephropathy, retinopathy, neuropathy, and diabetic heart failure.
  • the metabolic disease or disorder is obesity or obesity related complications.
  • a metabolic disease or disorder described in any embodiment herein is non-alcoholic fatty liver disease (NAFLD), comprising at least one prognosis stage of this disease selected from the group consisting of hepatic steatosis, nonalcoholic steatohepatitis (NASH), cirrhosis, and NAFLD induced hepatocellular carcinoma (HCC).
  • the metabolic disease or disorder is alcoholic fatty liver disease, or a complication caused by alcoholic fatty liver diseases.
  • the complication of alcoholic fatty liver disease comprises alcoholic hepatitis, cirrhosis, or a combination thereof.
  • the metabolic disease or disorder is dyslipidemia, or a complication caused by dyslipidemia.
  • the cancer is a primary cancer selected from the group consisting of hepatocellular carcinoma, colorectal carcinoma, pancreatic cancer, breast cancer, prostate cancer, leukemia, lymphoma, melanoma, ovarian cancer, and lung cancer.
  • the cancer is a metastatic cancer originated from a primary tumor of other tissue types.
  • the metastatic sites are selected from the group consisting of liver, lung and the intraperitoneal cavity.
  • the compound of embodiments described herein is administered in combination with a second agent indicated for the above-mentioned disorders or diseases, either concomitant with, prior to, or after the administration of the second agent.
  • the second agent is an anti-diabetic agent selected from the group consisting of metformin, insulin, insulin analogs, sulfonylureas, biguanides, meglitinides, thiazolidinediones, alpha glucosidase inhibitors, GLP-1 agonists, SGLT2 inhibitors and DPP-
  • the second agent is an anti-obesity agent. In some embodiments, the second agent is an anti -nonalcoholic fatty liver disease agent. In some embodiments, the second agent is anti-alcoholic fatty liver disease agent. In some embodiments, the second agent is an anti-dyslipidemia agent.
  • a compound of embodiments described herein is administered in combination with a second anti- non-alcoholic fatty liver disease agent. In some embodiments, a compound of the invention is administered in combination with a second anti- alcoholic fatty liver disease agent. In some embodiments, a compound of the invention is administered in combination with a second anti- dyslipidemia agent.
  • the compound may be administered in combination with a second anti-cancer agent or anti-cancer regimen.
  • the second anti-cancer agent s an immunoncological agent.
  • the immunocological agent is selected from the group consisting of an antibody against PD-1/PD-L1, an antibody against other immune check point proteins, CAR-T cells, and other therapeutic immune cells.
  • the compound may be administered prior to, concomitantly with, or subsequently to administration of the second anti-metabolic disease or anti-cancer agent.
  • the subject is a mammalian animal. In some embodiments, the subject is a human. In some embodiments, the compound described herein is used as a veterinarian drug to treat diabetes or a diabetes-associated disease, and the subject is a mammalian animal.
  • Some embodiments are directed to a method for long-term disease management of a metabolic disease or disorder comprising administering to a subject in need of such long-term management an effective amount of a compound or a pharmaceutical composition described herein.
  • a method for long-term disease management of a metabolic disease or disorder, or for long-term disease management of cancer comprises administering to a subject in need of such long-term management an effective amount of a compound or a pharmaceutical composition according to any of the embodiments described herein.
  • the metabolic disease or disorder is obesity, obesity -related complications, type 2 diabetes, or type 2 diabetes related complications.
  • the cancer is any primary tumor or metastatic tumor.
  • SUBSTITUTE SHEET ( RULE 26 ) accumulation in tissue is a symptom, or related disorders or complications, including, but not limited to, hepatic steatosis, non-alcoholic steatohepatitis (NASH), cirrhosis, or NAFLD induced hepatocellular carcinoma (HCC).
  • the compound of embodiments herein may be used to manufacture a medicament for the treatment of cancer, a disease where cell proliferation (hyperplasia) is a symptom, or cancer or hyperplasia related complications.
  • Some embodiments herein are directed to a method of treating or preventing a metabolic disease or disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein.
  • diseases or disorders may be caused by dietary, environmental, medical and/or genetic factors.
  • the methods described herein may also be used for prevention of the above-mentioned metabolic diseases for a subject with risk factors including, but not limited to, dietary, environmental, medical, and genetic predispositions.
  • some embodiments provide a method for long-term chronic disease management and longevity management by reducing insulin resistance or reducing glucose levels in the blood.
  • the metabolic disease or disorder is type 2 diabetes, or related diseases leading to insulin resistance or hyperglycemia. In some embodiments, the metabolic disease or disorder is obesity or one or more obesity related complications.
  • the metabolic disease or disorder is non-alcoholic fatty liver disease, (NAFLD), including nonalcoholic steatohepatitis (NASH) and cirrhosis, or alcoholic fatty liver disease (AFLD).
  • NAFLD non-alcoholic fatty liver disease
  • AFLD alcoholic fatty liver disease
  • the metabolic disease or disorder is non-alcoholic fatty liver disease, including nonalcoholic steatohepatitis (NASH) and cirrhosis, or alcoholic fatty liver disease (AFLD).
  • NASH nonalcoholic steatohepatitis
  • AFLD alcoholic fatty liver disease
  • SUBSTITUTE SHEET (RULE 26 ) is hepatic steatosis, non-alcoholic steatohepatitis (NASH), cirrhosis, or NAFLD induced hepatocellular carcinoma (HCC).
  • NASH non-alcoholic steatohepatitis
  • HCC NAFLD induced hepatocellular carcinoma
  • the cancer may be primary cancer or metastatic cancer.
  • the cancer is primary cancer including but not limited to hepatocellular carcinoma, colorectal carcinoma, pancreatic cancer, breast cancer, prostate cancer, leukemia, lymphoma, melanoma, ovarian cancer, lung cancer.
  • the cancer is metastatic liver cancer originated from the primary tumor of other tissue types.
  • the cancer is metastatic lung cancer originated from the primary tumor of other tissue types.
  • the cancer is metastatic cancer to other sites including intraperitoneal cavity.
  • Some embodiments are directed to a method of treating or preventing autoimmune diseases in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein.
  • the autoimmune disease is celiac disease, diabetes mellitus type 1, Graves' disease, inflammatory bowel disease, multiple sclerosis, psoriasis, rheumatoid arthritis, and systemic lupus erythematosus.
  • Some embodiments are directed to a method of treating or preventing a dermatological disorder in a subject in need thereof, comprising administering to the subject a
  • the dermatological disorder is eczema, dyshidrotic eczema, seborrheic eczema psoriasis, rosacea, dermatitis and atopic dermatitis.
  • Some embodiments are directed to a method of treating or preventing an infectious disease of non-viral parasites in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein.
  • the infectious disease is a viral infections.
  • the viral infection is an envelope viral infection.
  • the viral infection is selected from the group consisting of SARS- CoV-2 , a corana virus infection and an Ebola viral infection.
  • the disease to be treated may be a heart disorder comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein.
  • the heart disorder may be hypertension or cardiovascular disease.
  • the disease to be treated may be a central nervous system (CNS) disease.
  • the CNS disease may be stroke, Alzheimer’s, Parkinson’s, Huntington’s, or ALS (amyotropic lateral sclerosis).
  • the disease to be treated may be a disorder associated with increased ROS (reactive oxygen species) production comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein.
  • Increased ROS has been associated with aging, Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, ALS (amyotropic lateral sclerosis), mitochondrial diseases, and various cancers.
  • the compounds and pharmaceutical compositions described herein may be administered by oral, parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV, intracistemal injection or infusion, subcutaneous injection, or implant), by inhalation spray, ophthalmic, nasal, vaginal, rectal, sublingual, or topical routes of administration and may be formulated, alone or together, in suitable dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles appropriate for each route of administration.
  • the compounds and pharmaceutical compositions described herein may also be formulated as a controlled-release formulation.
  • compositions comprising an active ingredient and a dermatologically acceptable base and/or an ophthalmically
  • SUBSTITUTE SHEET ( RULE 26 ) acceptable base.
  • Such pharmaceutical compositions can be formulated, e.g., as solutions, suspensions, spray, lotions, gels, pastes, medicated sticks, balms, shampoos, soap bars, liquid soaps, creams or ointments.
  • the pharmaceutical composition is the form of an ointment that can be applied in or around the eye of a mammal, including a human.
  • a dermatologically and/or ophthalmically acceptable base includes a pharmaceutically acceptable ointment base.
  • suitable ointment bases include, but are not limited to oleaginous ointment bases such as petrolatum (e.g., liquid petrolatum or white petrolatum), plastibase, hard paraffin, white soft paraffin, yellow soft paraffin, liquid paraffin, emulsifying wax, microcrystalline wax, white bees wax, yellow bees wax, carnauba wax, wool wax (wool fat), mineral oil, olive oil, purified lanolin, anhydrous lanolin, and water soluble ointment bases such as polyethylene glycol (e.g., polyethylene glycol 400 or polyethylene glycol 3350), propylene glycol, polyoxyethylene, polyoxypropylene, or any combinations thereof.
  • polyethylene glycol e.g., polyethylene glycol 400 or polyethylene glycol 3350
  • propylene glycol polyoxyethylene, polyoxypropylene, or any combinations thereof
  • a dermatologically and/or ophthalmically acceptable base includes one or more polymers as suspending agents.
  • Useful polymers include, but are not limited to, water-soluble polymers such as cellulosic polymers, e g., hydroxypropyl methylcellulose, and water-insoluble polymers such as cross-linked carboxyl- containing polymers.
  • a dermatologically and/or ophthalmically acceptable base includes one or more viscosity enhancing agents.
  • suitable viscosity enhancing agents include, but are not limited to, methyl cellulose, xanthan gum, gum tragacanth, carboxymethyl cellulose, silica, silicone, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, hydroxypropylmethyl cellulose acetate stearate, hydroxypropylmethyl cellulose phthalate, carbomer, polyvinyl alcohol, alginates, acacia, chitosans, acacia, com starch, gelatin, or combinations thereof.
  • a dermatologically and/or ophthalmically acceptable base includes one or more dermatologically and/or ophthalmically acceptable pH adjusting agents or buffering agents, including, but not limited to, acids such as acetic, boric, citric, lactic, phosphoric and hydrochloric acids; bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium, lactate and tris-
  • acids such as acetic, boric, citric, lactic, phosphoric and hydrochloric acids
  • bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium, lactate and tris-
  • SUBSTITUTE SHEET ( RULE 26 ) hydroxymethylaminomethane; and buffers such as citrate/dextrose, sodium bicarbonate and ammonium chloride. Such acids, bases and buffers are included in an amount required to maintain pH of the composition in a dermatologically and/or ophthalmically acceptable range.
  • a dermatologically and/or ophthalmically acceptable base includes one or more dermatologically and/or ophthalmically acceptable salts in an amount required to bring osmolality of the composition into a dermatologically and/or ophthalmically acceptable range.
  • Such salts include, but are not limited to, those having sodium, potassium or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate or bisulfite anions; specific salts include, e.g., sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite, and ammonium sulfate.
  • a dermatologically and/or ophthalmically acceptable base includes one or more dermatologically and/or ophthalmically acceptable preservatives to inhibit microbial activity.
  • Suitable preservatives include, but are not limited to, mercury-containing substances such as merfen and thiomersal; stabilized chlorine dioxide; and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide and cetylpyridinium chloride.
  • a dermatologically and/or ophthalmically acceptable base includes one or more dermatologically and/or ophthalmically acceptable surfactants to enhance physical stability, or for other purposes.
  • Suitable nonionic surfactants include isohexadecane, cyclomethicone, copolymers of ethylene glycol and propylene glycol, polyoxyethylene fatty acid glycerides and vegetable oils, e.g., polyoxyethylene (60) hydrogenated castor oil; and polyoxyethylene alkylethers and alkylphenyl ethers, e.g., octoxynol 10, octoxynol 40.
  • a dermatologically and/or ophthalmically acceptable base includes one or more dermatologically and/or ophthalmically acceptable penetration enhancers to enhance physical stability, or for other purposes.
  • Penetration enhancers are substances which enhance passage of topically-applied compounds into the stratum, comeum of the skin and therefrom into the epidermis and dermis.
  • SUBSTITUTE SHEET (RULE 26 ) polysorbates, fatty acids (e g., oleic), bile salts, N-methylpyrrolidone, polyglycosylated glycerides, l-dodecylazacycloheptan-2-one (Azone®), Cyclopentadecalactone (CPE-215®), Alkyl -2-(N,N-disubstituted amino)-alkanoate ester (NexAct®), 2-(n-nonyl)- 1,3 -di oxolane (DEP A®), and penetration enhancers shown for example in U.S. Pat. Nos.
  • a dermatologically and/or ophthalmically acceptable base includes one or more dermatologically and/or ophthalmically acceptable permability enhancers to enhance physical stability, or for other purposes.
  • a variety of classes of compounds may serve as suitable permeability enhancers according to the invention.
  • a first category includes fatty acids and salts and esters thereof, including mono-, di-, and triglycerides. Medium chain length fatty acids, especially C8 and CIO acids, and their salts and esters are particularly useful.
  • Suitable specific examples include sodium caprylate, sodium caprate, CAPMUL® glycerides (available from Abitec of Columbus, Ohio), LABRASOL® glycerides (PEG-8 caprylic/capric glycerides, available from Gattefosse SAS of Saint Priest, Cedex, France), GELUCIRE® 44/14 (PEG-32 glyceryl laurate EP, available from Gattefosse), other glycerides & fatty acid esters, CREMOPHOR® (BASF, Ludwigshafen, Germany), D-a-tocopheryl polyethylene glycol 1000 succinate, vegetable oils, polyoxylglycerides, and medium chain mono- and diacylglycerides.
  • GATTEFOSSE compositions 61 A through 61H which are proprietary to Gattefosse SAS, but generally are composed of mixtures containing one or more of medium chain mono-, di-, or triglycerides, polysorbate derivatives, polyoxyl castor oil derivatives, polyethylene glycol derivatives including polyethylene glycol glycerides, polyoxyl ethers, vegetable oils, glycerin, and similar GRAS (generally regarded as safe) lipidic components in varying amounts.
  • CAPRYOLTM 90 CAPRYOLTM PGMC
  • LAUROGLYCOLTM 90 GELUCIRE® 44/14
  • Plurol Oleique CC497 LABRASOL®
  • LABRAFIL® M1944CS apricot kernel oil PEG-6 esters
  • Transcutol HP Peceol
  • Maisine 35-1 all of which are available from Gattefosse SAS.
  • a second category of enhancers includes surfactants having a steroidal structure, such as bile acid salts.
  • suitable compounds include sodium cholate, sodium deoxycholate, glycocholate, glycoursodeoxycholate, taurocholate, taurodeoxycholate, and steroid detergents/bile salts.
  • Other surfactants may also be suitable permeability enhancers, including cationic, anionic, and nonionic surfactants.
  • Examples include polysorbate 80, hexadecyldimethylbenzylammonium chloride, N-hexadecylpyridinium bromide, dodecyltrimethylammonium bromide, hexadecyltrimethylammonium bromide, tetradecyl-P-D-maltoside, octylglucoside, glycyrrhetinic acid, 3-(N,N- dimethylpalmitylammonio)propane-sulfonate, and sodium lauryl sulfate.
  • Cyclodextrins may also be used as suitable enhancers. Examples include P-cyclodextrin, hydroxypropyl-P-cyclodextrin, y-cyclodextrin, and hydroxypropyl-y- cyclodextrin.
  • the permeability enhancer and the polar agent may be mixed in any proportion so long as there is provided a therapeutically effective amount of the polar agent and a permeability-enhancing amount of the enhancer compound. Enhancement in dermal bioavailability of topically administered polar agents can depend on the nature and
  • SUBSTITUTE SHEET ( RULE 26 ) concentration of the enhancer compound with which the agent is formulated. It is thus contemplated that the required therapeutic amount may be contained in a single dosage form or divided between one or more dosages intended for application at the same time or in sequence.
  • the permeability enhancers act relatively independently of the concentration of polar agent. Differing permeability enhancers can reach either optimal or maximum enhancement over a wide concentration range depending on their particular inherent enhancement potential. Often, enhancers have a non-linear dose response relationship between concentration of enhancer present and amount of increased polar agent absorption.
  • the amount of enhancer to be utilized in an oral dosage form with a polar agent is initially based upon the enhancement properties observed in Caco-2 cell assays at varying fixed enhancer concentrations. Based upon those results, an effective in vivo amount of enhancer compound for a human formulation can be estimated, demonstrated and optimized without undue experimentation using methods well known to those skilled in the formulation art, to achieve a desired pharmacokinetic in vivo profile.
  • the amount of enhancer may be at least about 0.1 wt % of the combined weight of enhancer and polar agent, more preferably at least about 50 wt %, and more preferably at least 70 wt % of the combined weight of enhancer and polar agent.
  • the amount is preferably at most 95 wt %, more preferably at most 80 wt %, and more preferably at most 75 wt % of the combined weight of the enhancer and polar agent.
  • a typical dosage form may contain a wide range of concentrations of enhancer compounds depending on the compound itself and its efficacy in enhancing the permeability of polar agents following oral administration. Concentrations as low as 0.001% by weight up to 20% have been demonstrated to be effective in enhancement of the permeability of polar agents.
  • a dermatologically and/or ophthalmically acceptable base includes one or more antioxidants to enhance chemical stability where required.
  • Suitable antioxidants include, by way of example only, butylated hydroxytoluene (BHT), sodium ascorbate, ascorbic acid, sodium metabisulfite, and tocopherol.
  • antioxidants enhance chemical stability where required.
  • any other surfactant, moisturizer, gelling agent, preservative, colorant or pigment, antioxidant, radical scavenger, emulsifier, humectant, pH modifier, chelating agent, or other dermatologically acceptable excipient commonly known to those of ordinary skill in the art as useful in topical compositions is contemplated as useful in the compositions described herein.
  • any non-toxic, inert, and effective topical carrier may be used to formulate the compositions described herein.
  • Food and Drug Administration (FDA) Center for Drug Evaluation and Research (CDER) Office of Management, http://www.accessdata.fda.gov/scripts/cder/iig/index.cfm, the contents of which are hereby incorporated by reference in their entirety.
  • useful pharmaceutically acceptable excipients, carriers and diluents include distilled water, physiological saline, Ringer's solution, dextrose solution, Hank's solution, and DMSO, which are among those preferred for use herein.
  • composition may be used immediately or stored for later use in any type of container known to one of skill in the art such as, for example, pouch, jar, bottle, tube, ampule and pre-filled syringe. Finally, the composition may be sterilized by any method known to one of skill in the art such as, for example, y radiation.
  • compositions described herein may be administered at prophylactically effective dosage levels to prevent the above-recited conditions and disorders, as well as to prevent other conditions and disorders characterized by insulin resistance or hyperglycemia.
  • compositions and compounds of embodiments herein can be administered in a wide range of dosage-forms including, for example, solid dosage
  • Solid dosage forms may include powders, tablets, pills, capsules, suppositories, or dispersible granules.
  • a solid carrier can be one or more substances that function as a diluting agent, flavor additive, solvent, lubricant, suspension agent, binder, preservative, tablet-disintegrating substance or encapsulating material.
  • the carrier may be a finely pulverized solid including lactose, hydroxypropylmethylcellulose and PVP, mixed with an appropriate amount of the active ingredient.
  • Appropriate carriers for powder and tablet forms include for example magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, stiffeners, gelatins, tragacanth, methylcellulose, and sodium carboxymethylcellulose.
  • Liquid dosage forms include for example solutions, suspensions, and emulsions. Also included are pharmaceutical compositions in solid form that are meant to be converted to liquid form shortly prior to consumption. These forms may include, in addition to the active ingredients, artificial colors, flavors, stabilizers, buffers, natural or artificial sweeteners, dispersing agents, thickeners, dissolving agents and the like.
  • Solutions or mixtures may be administered directly to the nasal cavity using conventional means, such as drops or sprays.
  • the pharmaceutical composition may be produced in individual or multi-dose forms. Multi-dose forms would include a dropper, pipette or atomizer that delivers a predetermined volume of the pharmaceutical composition.
  • the pharmaceutical compositions and compounds of embodiments herein may be provided in individual dosage units that contain a suitable amount of the active ingredient.
  • the individual doses may be provided in a package, or as a kit that includes a measuring device, e.g., a device for measuring oral or injectable dosages (i.e., a measuring cup, needle, or syringe).
  • the kit can also include, other materials such buffers, diluents, filters, and package inserts with instructions for use.
  • a label may be present on the on the kit to indicate that the pharmaceutical composition is used for a specific therapy, and may also indicate directions for use.
  • the pharmaceutical compositions of the present invention may further comprise one or more additional active agents.
  • any of the active agents may be administered in the form of the compound per se, and/or in the form of a salt, polymorph, ester, amide, prodrug, derivative, or the like, provided the salt, polymorph, ester, amide, prodrug or derivative is suitable pharmacologically.
  • salts, esters, amides, prodrugs and other derivatives of the active agents may be prepared using standard procedures known to those skilled in the art of synthetic organic chemistry and described, for example, by J. March, Advanced Organic Chemistry: Reactions, Mechanisms
  • the active agent may be incorporated into the present pharmaceutical compositions either as the racemate or in enantiomerically enriched form.
  • the dosage of the active compound(s) being administered will depend on the condition being treated, the particular compound, and other clinical factors such as age, sex, weight, and health of the subject being treated, the route of administration of the compound(s), and the type of pharmaceutical composition being administered (tablet, gel cap, capsule, solution, suspension, inhaler, aerosol, elixir, lozenge, injection, patch, ointment, cream, etc.) It is to be understood that the present disclosure has application for both human and animal use. The amount of the compound, or an active salt or derivative thereof, required for use in treatment will be ultimately at the discretion of the attendant physician or clinician.
  • the compounds of the invention are useful for the prevention, treatment, control, amelioration, or reduction of risk of the diseases, disorders and conditions noted herein.
  • the dosage of the compound as an active ingredient in the pharmaceutical compositions of this invention may be varied so that a suitable dosage form is obtained.
  • the active ingredient may be administered to patients (animals and human) in need of such treatment in dosages that will provide optimal pharmaceutical efficacy.
  • the selected dosage depends upon the desired therapeutic effect, on the route of administration, and on the duration of the treatment.
  • the dose will vary from patient to patient depending upon the nature and severity of disease, the patient's weight, special diets then being followed by a patient, concurrent medication, and other factors which those skilled in the art will recognize. Generally, dosage levels of between 0.001 to 100 mg/kg.
  • the therapeutically effective amount will generally be about 0.5 mg to 10g per patient per day which may be administered in single or multiple doses. In some embodiments the therapeutically effective amount is between a lower limit of 0.5 mg, 10 mg, 1 mg, 500.0 mg, 1000 mg, 1500 mg, 2000 mg, 2500 mg, 3000 mg, 3500 mg, 4000 mg, 4500 mg, 5000 mg, 5500 mg, 6000 mg, 6500 mg, 7000 mg, 7500 mg, 8000 mg, 8500 mg, 9000 mg, 9500 mg, and 10000 mg; and an upper limit of 10000 mg, 9500 mg, 9000 mg, 8500 mg, 8000 mg, 7500 mg, 7000 mg, 6500 mg, 6000 mg, 5500 mg, 5000 mg, 4500 mg, 4000 mg, 3500 mg, 3000 mg, 2500 mg, 2000 mg, 1500 mg, 1000 mg, 500.0 mg, 100 mg, 10 mg and 0.5 mg. In some embodiments, the therapeutically
  • SUBSTITUTE SHEET ( RULE 26 ) per patient per day; in some embodiments about 0.5 mg to 1000 mg per patient per day; and in yet some other embodiments about 5 mg to 50 mg per patient per day.
  • Pharmaceutical compositions of the present invention may be provided in a solid dosage formulation such as comprising about 0.5 mg to 500 mg active ingredient, or comprising about 1 mg to 250 mg active ingredient.
  • the pharmaceutical composition may be provided in a solid dosage formulation comprising for example about 1 mg, 5 mg, 10 mg, 25 mg, 50 mg, 100 mg, 200 mg, 250 mg, 500 mg or 1000 mg of active ingredient.
  • acylamino denotes a nitrogen radical adjacent to an acyl group.
  • alkyl is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms.
  • C1-C17- alkyl or “C1.17 alkyl” (or alkylene)
  • Ci-Ce alkyl or "Ci-6 alkyl” denotes alkyl having 1 to 6 carbon atoms.
  • alkenyl is intended to include hydrocarbon chains of either straight or branched configuration having the specified number of carbon atoms and one or more, preferably one to three, carbon-carbon double bonds that may occur in any stable point along the chain.
  • C2-C6 alkenyl or “C2-6 alkenyl” (or alkenylene)
  • C2-17 alkenyl is intended to include C2, C3, C4, C5, Ce, C7, Cs, C9, C10, Cn, C12, C13, C14, C15, Ci6, and C17 alkenyl groups.
  • alkoxy refers to an -O-alkyl group.
  • Ci-Ce alkoxy or “Ci-6 alkoxy” (or alkyloxy)
  • alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), and t-butoxy.
  • aryl may be unsubstituted or substituted with 1 to 5 groups selected from -OH, -OCH3, -CI, -F, -Br, -I, -CN, -NO2, -NH 2 , -NH(CH 3 ), -N(CH 3 ) 2 , -CF 3 , -OCF3, -C(O)CH 3 , - SCH3, -S(O)CH 3 , -S(O) 2 CH3, -CH 3 , -CH2CH3, -CO2H, and -CO2CH3.
  • benzyl refers to a methyl group on which one of the hydrogen atoms is replaced by a phenyl group, wherein said phenyl group may optionally be substituted by one to five, preferably one to three, substituents independently selected from methyl, trifluoromethyl (-CF3), hydroxyl (-OH), methoxy (-OCH3), halogen, cyano (-CN), nitro (-NO2), -CO2Me, -CO2Et, and -CO2H.
  • a "compound,” as used herein, refers to any type of substance or agent that is commonly considered a drug, or a candidate for use as a drug, as well as combinations and mixtures of the above.
  • the term “compound” is intended to encompass not only the specified molecular entity but also its pharmaceutically acceptable, pharmacologically active analogs, including, but not limited to, salts, polymorphs, esters, amides, prodrugs, adducts, conjugates, active metabolites, and the like, where such modifications to the molecular entity are appropriate.
  • a “conventional mitochondrial uncoupler” as used herein describes a mitochondrial uncoupler that has properties that increase OCR and decreased MMP, and the concentrations for increasing OCR and dissipating MMP correlate.
  • a "derivative" of a compound refers to a chemical compound that may be produced from another compound of similar structure in one or more steps. Non-limiting examples include replacement of H by an alkyl, acyl, or amino group.
  • an "effective amount” or “therapeutically effective amount” means an amount sufficient to produce a selected effect, such as alleviating symptoms of a disease or disorder.
  • an effective amount of a combination of compounds refers collectively to the combination as a whole, although the actual amounts of each compound may vary.
  • the term "more effective” means that the selected effect is alleviated to a greater extent by one treatment relative to the second treatment to which it is being compared.
  • halo refers to fluoro, chloro, bromo, and iodo.
  • Haloalkyl is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, substituted with one or more halogens. Examples of haloalkyl include, but are not limited to, fluoromethyl, difluoromethyl, trifluorom ethyl, and tri chloromethyl.
  • heteroaryl is intended to mean stable monocyclic and polycyclic aromatic hydrocarbons that include at least one heteroatom ring member, such as sulfur, oxygen, or nitrogen.
  • Heteroaryl groups include, without limitation, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, furyl, quinolyl, isoquinolyl, thienyl,
  • SUBSTITUTE SHEET ( RULE 26 ) imidazolyl, thiazolyl, indolyl, pyrroyl, oxazolyl, benzofuryl, benzothienyl, benzthiazolyl, isoxazolyl, pyrazolyl, triazolyl, tetrazolyl, indazolyl, 1,2,4-thiadiazolyl, isothiazolyl, purinyl, carbazolyl, benzimidazolyl, indolinyl, benzodioxolanyl, and benzodioxane.
  • heteroaryl groups may be unsubstituted or substituted with 1 to 5 groups selected from -OH, -OCH 3 , -CI, -F, -Br, -I, -CN, -NO 2 , -NH 2 , -NH(CH 3 ), -N(CH 3 ) 2 , -CF 3 , -OCF 3 , - C(O)CH 3 , -SCH 3 , -S(O)CH 3 , -S(O) 2 CH 3 , -CH 3 , -CH 2 CH 3 , -CO 2 H, and -CO 2 CH 3
  • the nitrogen atom is substituted or unsubstituted (i.e., N or NR wherein R is H or another substituent, if defined).
  • the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., N— >0 and S(O) P , wherein p is 0, 1 or 2).
  • heterocyclyl is defined as a saturated or partially unsaturated ring containing one to four hetero atoms or hetero groups selected from O, N, NH, -N(R Z )-, -S(O)- or -S(O) 2 - , wherein R z is selected from alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, optionally substituted heterocyclyl, in a single or fused heterocyclic ring system having from three to twelve ring members.
  • a heterocyclyl is a ring system having three to seven ring members
  • a heterocyclyl group include, without limitation, azetindinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, morpholinyl, dioxothiomorphylinyl, tetrahydrofuranyl and azabicyclo[3.2.1]octanyl.
  • heteroaryl groups may be unsubstituted or substituted with at least one groups selected from oxo, cyano, hydroxyl, alkoxy, -acylamino , carboxyamido, -SO 2 CH 3 , -CF 3 , Ci-Ce alkyl, halo, and acyl.
  • infectious disease referes to a bacterial infection, or viral infection. Infectious diseases do not include infections caused by parasitic organism.
  • the viral infection is an envelope virus. Examples of envelope viruses include SARS-CoV-2 , corana viruses and Ebola viruses.
  • mitochondrial malfunction is defined as pathologic conditions caused by mitochondrial malfunction as reviewed and summarized in: A Mitochondrial Paradigm of Metabolic and Degenerative Diseases, Aging, and Cancer: A Dawn for Evolutionary Medicine, Annu Rev Genet. 2005; 39: 359, The rise of mitochondria in medicine, Mitochondrion 2016, 30:105-16, and Is Mitochondrial Dysfunction a Common Root of Noncommuni cable Chronic Diseases? Endocrine Reviews 2020,41(491-517), all of which are incorporated herein by reference. These conditions include, but are not limited to: genetic mitochondrial diseases, various types of cancer, autisim, neurodegenerative diseases, neuromuscular diseases, immunological diseases, metabolic diseases, aging, and aging- related noncommuni cable chronic diseases.
  • mitochondrial uncoupling also referred to as “uncoupling” refers to the process whereby protons enter the mitochondrial matrix via a pathway independent of ATP synthase and thereby uncouple nutrient oxidation from ATP production.
  • This process can be pharmacologically induced by small molecule mitochondrial protonophores, which directly shuttle protons across the mitochondrial inner membrane into the matrix.
  • the primary pathway for energy production in aerobic cells involves the oxidation of nutrients (including fats, carbohydrates, and amino acids) in mitochondria, which promotes the efflux of protons out of the mitochondrial matrix. This process creates a pH and electrochemical gradient across the mitochondrial inner membrane.
  • Protons normally re-enter the mitochondrial matrix via ATP synthase, which results in ATP production. Protons can also re-enter the mitochondrial matrix via pathways independent of ATP synthase, which 'uncouples' nutrient oxidation and proton efflux from ATP production.
  • opthalmically acceptable is employed herein to refer to those compounds, materials, pharmaceutical compositions, and/or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the eyes of human beings and animals without excessive toxicity, irritation, allergic response, and/or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • SUBSTITUTE SHEET (RULE 26 ) acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, and isethionic.
  • acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane
  • the term "pharmaceutically acceptable carrier” includes any of the standard pharmaceutical carriers, such as a phosphate buffered saline solution, water, emulsions such as an oil/water or water/oil emulsion, and various types of wetting agents.
  • the term also encompasses any of the agents approved by a regulatory agency of the US Federal government or listed in the US Pharmacopeia for use in animals, including humans.
  • a "prodrug” refers to an agent that is converted into the parent drug in vivo. Prodrugs are often useful because, in some situations, they may be easier to administer than the parent drug. They may, for instance, be bioavailable by oral administration whereas the parent is not. The prodrug may also have improved solubility in pharmaceutical compositions over the parent drug, or may demonstrate increased palatability, or be easier to formulate.
  • treating refers to administration of a compound or agent to a subject who has a disorder or is at risk of developing the disorder with the purpose to cure, alleviate, relieve, remedy, delay the onset of, prevent, or ameliorate the disorder, the symptom of the disorder, the disease state secondary to the disorder, or the predisposition toward the disorder.
  • Example 2 0.33 mmol, Example 2 was dissolved in DCM (3.0 mL), followed by the addition of catalytic amount of DMF (10 pL) and oxalyl chloride (34 pL, 0.39 mmol) respectively. The reaction was allowed to stir at rt for 30 minutes, concentrated in vacuo, and the 5-chloro-2- methoxy-3-((2-methoxyethoxy)methyl)benzoyl chloride residue was re-dissolved in THF (3.0 mL).

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Abstract

The present application discloses a distinct class of mitochondrial uncoupling compounds as well as conventional mitochondrial uncoupling compounds with drastically improved pharmacokinetic properties, pharmaceutical compositions containing the compounds and/or the prodrugs and methods of using the compounds, prodrugs and pharmaceutical compositions in treating diseases related to mitochondrial dysfunctions or diseases benefiting from modulating mitochondrial activities, including diabetes, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), mitochondrial genetic diseases, neurodegenerative diseases, cancer, autoimmune diseases, and infectious diseases.

Description

MITOCHONDRIAL UNCOUPLERS FOR TREATMENT OF METABOLIC DISEASES AND CANCER
CROSS REFERENCE TO RELATED APPLICATIONS
(00011 This application claims priority to U.S. Provisional Application No. 63/374,927, filed on September 8, 2022, which is hereby incorporated by reference in its entirety.
SUMMARY
[0002] The present disclosure relates to new mitochondrial uncoupling compounds that retain mitochondrial membrane potential as well as new mitochondrial uncoupling compounds with unexepctedly improved pharmacokinetic properties. Various embodiments described herein provide benzamide compounds, prodrugs of the compounds, pharmaceutical compositions containing the compounds and/or the prodrugs and methods of using the compounds, prodrugs and pharmaceutical compositions in the treatment of diseases related to metabolism including diabetes, Non-Alcoholic Fatty Liver Disease (NAFLD), Non- Alcoholic Steathohepatitis (NASH), cancer, autoimmune diseases, dyslipodemia, and infectious diseases.
[0003| Some embodiments of the present disclosure are directed to a compound of Formula A: wherein R1000a of Formula A is selected from the group consisting of
-CH3, -CH2CH3, -CI-CS alkyl, -C3-C6 cycloalkyl, -OCH3, -CH2OCH3, -CH2OCH2OCH3,
C(O)N(CH2CH2OCH3)2;
SUBSTITUTE SHEET ( RULE 26 ) each of substituents R5000A andR5000B is independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH2)2OCH3; alternatively R5000A and R5000B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl;
R6000 is selected from the group consisting of a 5 to 6-membered heterocyclyl, pyridinyl and thiazolyl; r’ is an integer selected the group consisting of 1, 2 and 3; r is an integer selected from the group consisting of 0, 1, 2 and 3;
Rloooc of Formula I is selected from the group consisting of chloro, fluoro, iodo, and bromo; each of R4000b and R4000d is independently selected from the group consisting of Y1000 and Z1000, provided that when R4000b is Y1000, R4000d is z1000 and when R4000b is z1000, R4000d is yiooo.
Y1000 is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , - CHF2, fluoro(Ci-C6)alkyl, halo(Ci-C6)alkyl, -OCF3, -SO2(Ci-C6)alkyl, cyano, and -CO2(Ci- Ce)alkyl;
Z1000 is selected from the group consisting of H, -CH2OCH3, -CH2OCH2CH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3, -CH2O(CH2)2N(CH3)2, -CH2O(CH2)2NHSO2CH3, -(CH2)O(CH2)2NR2000AR2000B, -(CH2)SR3000, -CI FOCI F Ar1, OCH3 CH2NHC(O)CH2CH3, -CH2NHC(O)CH2OCH3, -CH2NHSO2CH3, -(CH2)t’NR7000AR7000B, and -(CH2)tR8000;
R2OOOA anj R2000B t|ie nitrogen to which they are attached, form a 4 to 8- membered heterocyclic ring optionally substituted with one or more methyl groups;
R3000 is a 5 to 6-membered heterocyclic ring;
Ar1 is a 5 to 6-membered aryl or heteroaryl group optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl, halo, hydroxyl and alkoxy;
-2-
SUBSTITUTE SHEET ( RULE 26 ) each of R7000A and R7000B is independently selected from Ci-Ce alkyl; alternatively, R7OOOA anj R7000B tOgether with the nitrogen to which they are attached, form a 4 to 8- membered heterocyclyl optionally substituted with one or more substituents independently selected from Ci-Ce alkyl;
R8000 is selected from the group consisting of a 5 to 6-membered heterocyclic ring optionally substituted with methyl; s is an integer selected from the group consisting of 0, 1, 2 and 3; t’ is an integer selected from the group consisting of 1, 2 and 3; t is an integer selected from the group consisting of 0, 1, 2 and 3; with the proviso that
R5000Aand R5000B are not both Ci-Ce alkyl; and when Z1000 is H; R1000a is not Ci- C6 alkyl, -C3-C6 cycloalkyl, CH3 or CH2CH3; or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
[0004] Some embodiments of the present disclosure are directed to a compound of Formula I: wherein:
Rla is selected from the group consisting of -CH3, -CH2CH3, -OCH3, -CH2OCH3, -CH2OCH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3, -(CH2)NH(CH2)2OCH3,
SUBSTITUTE SHEET ( RULE 26 ) each of R5A andR5B is independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH2)2OCH3; alternatively R5A and R5B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino, carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl;
R6 is selected from the group consisting of a 5 to 6-membered heterocyclyl, pyridinyl and thiazolyl; m’ is an integer selected the group consisting of 1, 2 and 3; m is an integer selected the group consisting of 0, 1, 2 and 3;
Rlc is selected from the group consisting of chloro, fluoro, iodo, and bromo; each of R4b and R4d is independently selected from the group consisting of Y and Z, provided that when R4b is Y, R4d is Z and when R4b is Z, R4d is Y;
Y is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-C6)alkyl, halo(Ci-C6)alkyl, -OCF3, -SO2(Ci-C6)alkyl, cyano, and -CO2(Ci-C6)alkyl;
Z is selected from the group consisting of -CH2OH, -CH2OCH2CH3, -CH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3, -CH2O(CH2)2NHCH3J -CH2O(CH2)2N(CH3)2J -CH2O(CH2)2NHSO2CH3J -(CH2)O(CH2)2NR2AR2B, -(CH2)nR3, -CH2OCH2Ar, and -OCH3;
R2A and R2B together with the nitrogen to which they are attached, form a 4 to 8- membered heterocyclic ring optionally substituted with one or more methyl groups;
R3 is selected from the group consisting of a 5 to 6-membered heterocyclic ring and phenoxy; n is an integer selected from the group consisting of 0 1, 2 and 3; and
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SUBSTITUTE SHEET ( RULE 26 ) Ar is a 5-6 membered aryl or heteroaryl group optionally substituted with one or more substituents, independently selected from the group consisting of -Ci-Ce alkyl, halo, hydroxy, and alkoxy; or a pharmaceutically acceptable salt, solvate, or prodrug thereof
[0005] Some embodiments of the present disclosure are directed to a compound of Formula II: wherein:
R10a is selected from the group consisting of -OCH3, -CFFOCHa, -CH2OCH2OCH3,
C(O)N(CH2CH2OCH3)2; each of R50AandR50B is independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH2)2OCH3; alternatively R50A and R50B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl;
R60 is selected from the group consisting of 5 to 6-membered heterocyclyl, pyridinyl and thiazolyl; o’ is an integer selected from the group consisting of 1, 2 and 3; o is an integer selected from the group consisting of 0, 1, 2 and 3;
R10c is selected from the group consisting of chloro, fluoro, iodo, and bromo; and
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SUBSTITUTE SHEET ( RULE 26 ) one of R40b and R40d is H and the other of R40b and R40d is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-Ce)alkyl, halo(Ci-Ce)alkyl, -OCF3, -SO2(Ci-C6)alkyl, cyano, and -CO2(Ci-C6)alkyl; with the proviso that R50A and R50B are not both Ci-Ce alkyl; or a pharmaceutically acceptable salt, solvate, or prodrug thereof
100061 Some embodiments of the present disclosure are directed to a compound of Formula III: wherein:
R100a is selected from the group consisting of -CH3, -OCH3, -CH2OCH3, each of R500A and R500B is independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH2)2OCH3; alternatively R500A and R500B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl; R600 is selected from the group consisting of a 5 to 6-membered heterocyclyl, pyridinyl and thiazolyl; p’ is an integer selected the group consisting of 1, 2, and 3; p is an integer selected the group consisting of 0, 1, 2 and 3;
SUBSTITUTE SHEET ( RULE 26 ) R100c is selected from the group consisting of chloro, fluoro, iodo, and bromo; each of R400b and R400d is independently selected from the group consisting of Y1 and Z1, provided that when R400b is Y1, R400d is Z1 and when R400b is Z1, R400d is Y1;
Y1 is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-C6)alkyl, halo(Ci-C6)alkyl, -OCF3, -SO2(Ci-C6)alkyl, cyano, and -CO2(Ci-C6)alkyl;
Z1 is selected from the group consisting of -CH2NHC(O)CH2CH3, -CH2NHC(O)CH2OCH3, -CH2NHSO2CH3, -(CH2)q NR7AR7B, and -(CH2)QR8; each ofR7A andR7B is independently selected from Ci-Ce alkyl; alternatively R7A and R7B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more independently selected Ci-Ce alkyl;
R8 is selected from the group consisting of a 5 to 6-membered heterocyclic ring optionally substituted with methyl; q' is an integer selected from the group consisting of 1, 2, and 3; and q is an integer selected from the group consisting of 0 1, 2 and 3, or a pharmaceutically acceptable salt, solvate, or prodrug thereof
|0007] Embodiments herein describe a pharmaceutical composition comprising: a compound according to any embodiment described herein, or a pharmaceutically acceptable salt or prodrug thereof; and a pharmaceutically acceptable carrier or diluent.
10008] Some embodiments describe a method of treating a mitochondria-related condition or disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein. In some embodiments the mitochondria-related condition or disorder has and one or more underlying causal factors or symptoms selected from the group consisting of hyperglycemia, abnormal accumulation of lipid in cells, abnormal accumulation of lipid in tissues, abnormal lipid metabolism, insulin resistance, abnormal cell proliferation, abnormal TGF-beta activation, and abnormal fibrosis. In some embodiments, the mitochondria-related condition or disorder is selected from the group consisting of a metabolic disease, cancer, an
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SUBSTITUTE SHEET ( RULE 26 ) autoimmune disease, pulmonary fibrosis, a dermatological disorder, an infectious disease, and a neurodegenerative disease.
|0009] In some embodiments the metochondria-rel ted condition or disorder is a metabolic disease selected from the group consisting of type 2 diabetes, a disease characterized by insulin resistance or hyperglycemia; obesity or obesity related complications, and a disease characterized by abnormal lipid accumulation. In some embodiments the metabolic disease is a complication caused by type 2 diabetes, selected from the group consisting of diabetes-induced cardiovascular diseases, neurodegenerative disorders, atherosclerosis, hypertension, coronary heart diseases, nephropathy, retinopathy, neuropathy and diabetic heart failure. In some embodiments the metabolic disease or disorder is non-alcoholic fatty liver disease (NAFLD), wherein at least one prognosis stage of the disease is selected from the group consisting of hepatic steatosis, non-alcoholic steatohepatitis (NASH), cirrhosis, and NAFLD induced hepatocellular carcinoma (HCC). In some embodiments the metabolic disease or disorder is alcoholic fatty liver disease, or one or more complications caused by alcoholic fatty liver disease, wherein the one or more complications caused by alcoholic fatty liver disease is selected from the group consisting of alcoholic hepatitis, cirrhosis, and a combination thereof. In some embodiments the metabolic disease or disorder is dyslipidemia, or one or more complications caused by dyslipidemia. In some embodimets the pharmaceutical composition is administered in combination with a second agent indicated for the metabolic disease. In some embodiments the second agent is an anti-diabetic agent selected from the group consisting of metformin, insulin, insulin analogs, sulfonylureas, biguanides, meglit-inides, thiazolidinediones, alpha glucosidase inhibitors, GLP-1 agonists, DPP-4 inhibitors and SGLT2 inhibitors. In some embiodiments the second agent s selected from the group consisting of an anti-obesity agent, an anti- nonalcoholic fatty liver disease agent, an anti-nonalcoholic fatty liver disease agent and an anti-dyslipidemia agent.
[0010] In some embodiments the metochondria-related condition or disorder is a primary cancer selected from the group consisting of hepatocellular carcinoma, colorectal carcinoma, pancreatic cancer, breast cancer, prostate cancer, leukemia, lymphoma, melanoma, ovarian cancer, and lung cancer. In some embodiments the cancer is a metastatic cancer originated from a primary tumor of other tissue types. In some mebodiments the pharmaceutical composition is administered in combination with a second anti-cancer agent or anti-cancer regimen. In some emodiments the second anti-cancer agent is an immunoncological agent. In some embodiments the immunocological agent is selected from
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SUBSTITUTE SHEET ( RULE 26 ) the group consisting of an antibody against PD-1/PD-L1, an antibody against other immune check point proteins, CAR-T cells, and other therapeutic immune cells.
10011] In some embodiments the metochondria-related condition or disorder is a dermatological disorder selected from eczema, dyshidrotic eczema, seborrheic eczema psoriasis, rosacea, dermatitis and atopic dermatitis.
]0012| In some embodiments the metochondria-related condition or disorder is an infectious disease. In some embodiments the infectious disease is a bacterial infection. In some embodiments the infectious disease is viral infection. In some embodiments the viral infection is selected from SARS-CoV-2 infection, a corona viral infection, and Ebola viral infection.
[0013] Some embodiments describe a method of treating a metabolic disease or disorder characterized by hyperglycemia, or insulin resistance or by abnormal accumulation of lipid in tissue, a disease or a disorder in which hyperglycemia, or insulin resistance or abnormal accumulation of lipid in tissue is a symptom, in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition described herein.
10014] Some embodiments describe a method of treating cancer or hyperplasia, in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition described herein.
[0015] Some embodiments describe a method of treating fibrosis, in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition described herein.
[0016] . Some embodiments describe a method of treating or preventing autoimmune diseases in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of oa pharmaceutical composition described herein.
|0017] Some embodiments describe a method of treating or preventing a dermatological disorder, in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition described herein.
[0018] Some embodiments describe a method of treating fibrosis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a a pharmaceutical composition described herein.
[0019] Some embodiments describe a method of treating or preventing a bacterial infection in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition described herein.
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SUBSTITUTE SHEET ( RULE 26 ) [0020| Some embodiments describe a method of treating or preventing viral infection in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition described herein
[0021] Some embodiments herein describe methods wherein the subject is a mammal or a human. In some embodiments, the subjet is a human.
[0022| Some embodiments herein describe methods wherein the pharmaceutical composition is administered orally, intravenously, subcutaneously, intramuscularly, transdermally, intraperitoneally, or by other pharmacologically acceptable routes.
[0023] Some embodiments describe a method for long-term disease management of a metabolic disease or disorder, or for long-term disease management of cancer, comprising administering to a subject in need of such long-term management an effective amount of a compound or a pharmaceutical composition described herein.
[0024] Some embodiments describe the use of a compound described herein in the manufacture of a medicament for treatment of a metabolic disease or disorder. In some embodiments, the metabolic disease or disorder may be selected from diabetes, obesity, nonalcoholic fatty liver disease, alcoholic fatty liver disease, dyslipidemia, a disease where hyperglycemia, or insulin resistance or abnormal lipid accumulation in tissue is a symptom, or related disorders or complications.
[0025] Some embodiments describe a method of preparing a mitochondrial membrane potential (MMP)-retaining mitochondrial uncoupler comprising:
1. identifying a conventional mitochondrial uncoupler;
2. designing a compound that covalently links at least one secondary or tertiary amino moiety to a conventional mitochondrial uncoupler; and
3. preparing the compound of step 2, wherein the compound is a mitochondrial membrane retaining uncoupler compound.
[0026] Some embodiments describe a mitochondrial membrane-retaining uncoupler compound of the Formula:
(RA)U-RB; or a pharmaceutically acceptable salt, solvate or prodrug thereof; wherein RAand RB are covalently linked; each RA is independently a moiety containing a secondary or tertiary amine;
-10-
SUBSTITUTE SHEET ( RULE 26 ) u is an integer selected from the group consisting of 1 and 2; and
RB is a conventional mitochondrial uncoupler prior to being covalently linked to RA; provided the mitochondrial membrane-retaining uncoupler compound is not
DESCRIPTION OF THE DRAWINGS
[0027] Figure 1 shows a schematic diagram illustrating the role of mitochondrial functions and dysfunctions in various diseases, including obesity, T2D (type 2 diabetes), NASH (non-alcoholic steatohepatitis), viral infection, cancer, cancer immune suppression (Cancer ME, cancer microenvironment), and neurodegenerative diseases. Mitochondrial uncoupling, by reducing metabolic output of metabolites and ROS, would help eliminate the causal factors of these diseases.
[0028] Figure 2 shows oxygen consumption rates (OCR) over time of two representative compounds. Compound 16 (Figure 2A) and Compound 69 (Figure 2B). Oligo represents oligomycin, Comp, A represents Compound 16, Comp, B represents compound 69, and Rot/ A A represents Rotenone/ Antimycin. The assay was performed with Seahorse XF-24 instrument. The compounds were injected into assay media in the order as indicated, Oligomycin at 2.5 pM; each of Comp. A and Comp.B at a concentration at 1.0 pM; Rotenone at 2.0 pM and Antimycin A at 2.0 pM.
[0029] Figure 3 shows the molecular mechanism of conventional mitochondrial uncoupling. Dissipating mitochondrial membrane potential is an intrinsic property for
-11-
SUBSTITUTE SHEET ( RULE 26 ) conventional mitochondrial uncouplers. The mitochondrial outer membrane, inner membrane, electron transport complexes I- IV, ATP synthase, a chemical uncoupler (U-, UH), and mitochondrial matrix are shown as indicated. Conventional uncouplers are weak lipophilic weak acid localized in mitochondrial inner membrane (shown as the deprotonated form U" and protonated form UH). U" binds to a proton at the outer side of the mitochondrial inner membrane (protonation) and releases the proton into the mitochondrial matrix (deprotonated). As a result, the mitochondrial uncoupler catalyzes proton translocation across the inner membrane without ATP synthesis, leading to 'futile' oxidation of acetyl-CoA (an end metabolic product of lipid oxidation and glucose metabolism). As the MMP is established by the proton gradient across the mitochondrial inner membrane, conventional mitochondrial uncoupling is associated with a simultaneous MMP dissipation.
(0030] Figure 4 shows the effect of a a representative conventional mitochondrial uncoupler, trifluoromethoxy carbonylcyanide phenylhydrazone (FCCP), on oxygen consumption rate and mitochondrial membrane potential. FCCP simultaneously increases oxygen consumption rate (Figure 4A) and dissipates mitochondrial membrane potential (Figure 4B). Figure 4A shows cellular oxygen consumption rates (OCR), determined by Seahorse OCR assay using C2C12 cells. The diamond in Figure 4A represents DMSO (vehicle treatment control), the triangle represents FCCP at 3.0 pM, the square represents FCCP at 6.0 pM, the X represents FCCP at 12.0 pM, the * represents FCCP at 18.0 pM, and the circle represents FCCP at 24 pM. Oligo is oligomycin 2.5 pM; AA isantimycin A 2 pM; Rot isrotenone 2 pM. Figure 4B shows mitochondrial membrane potential (MMP) determined by fluorescence microscopy using two different dyes, tetramethylrhodamine ethyl ester (TMRE) staining and DilCl(5) staining, respectively, in the presence of FCCP at the indicated concentrations. The fluorescent intensity is indicative of MMP. FCCP has a ratio of CIO%TMRE / Cmin-OCR less than 3, where CIO%TMRE is the concentration leading to 10% MMP retention (or 90% MMP loss, measured by TMRE staining) and Cmin-ocR is the minimal concentration leading to OCR increase.
[0031] Figure 5 shows the effect of a representative MMP -retaining mitochondrial uncoupler, Compound 25 (#25), on oxygen consumption rate and mitochondrial membrane potential. Compound 25 effectively uncouples mitochondria (induces OCR, Figure 5 A) without reducing MMP (Figure 5B). Figure 5A shows cellular oxygen consumption rates determined by Seahorse OCR assay using C2C12 cells. The diamond in Figure 5A represents vehicle treatment control, the square represents Compound 25 (#25) at 1.0 pM, the triangle represents Compound 25 at 3.0 pM, the X represents Compound 25 at 6.0 pM, the *
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SUBSTITUTE SHEET ( RULE 26 ) represents Compound 25 at 9.0 pM, and the circle represents Compound 25 atl2.0 pM. Oligo, isoligomycin 2.5 pM; AA is antimycin A 2 pM; Rot isrotenone 2 pM. Figure 5B shows mitochondrial membrane potential (MMP) determined by fluorescence microscopy using two different dyes, tetramethylrhodamine ethyl ester (TMRE) staining and DilCl(5) staining, respectively, in the presence of Compound 25(#25) at the indicated concentrations. The fluorescent intensity is indicative of MMP. Compound 25 represents the MMP -retaining uncouplers that do not cause observable MMP reduction while uncoupling mitochondria (with a ratio of CIO%TMRE I Cmin-ocR greater than 25).
[0032] Figure 6 shows the effect of Compound 64 (#64), on oxygen consumption rate (Figure 6A) and mitochondrial membrane potential (Figure 6B). Compound 64 increases OCR without drastically dissipating mitochondrial membrane potential. Figure 6A shows cellular oxygen consumption rates determined by Seahorse OCR assay using C2C12 cells at the indicated concentrations. The diamond in Figure 6A represents vehicle treatment control, the square represents Compound 64 (#64) at 0.3 pM, the triangle represents Compound 64 at 1.0 pM, the X represents Compound 64 at 2.0 pM, the * represents Compound 64 at 3.0 pM, and the circle represents Compound 64 at 4.0 pM. Oligo is oligomycin 2.5 pM; AA is antimycin A 2 pM; Rot is rotenone 2 pM. Figure 6B shows mitochondrial membrane potential (MMP) determined by fluorescence microscopy using two different dyes, tetramethylrhodamine ethyl ester (TMRE) staining and DilCl(5) staining, respectively, in the presence of Compound 64 (Comp #64) at the indicated concentrations. The fluorescent intensity is indicative of MMP. Compound 64 represents the MMP-retaining uncouplers that do not drastically reduce MMP while uncoupling mitochondria (with a ratio of CIO%TMRE / Cmin-ocR between 10 and 25).
[00331 Figure 7 shows a schematic illustration of the mechanism of action of a conventional mitochondrial uncoupler (Figure 7A) and a proposed mechanism of action of an MMP-retaining uncoupler (Figure 7B), as well as diagrams and an example of how to convert a conventional uncoupler to an MMP-retaining uncoupler (Figure 7C-E). Figure 7A and Figure 7B show mitochondrial electron transport chain complexes I, II, III, IV, ATP synthase, as well as uncouplers (UH or U"). Conventional uncouplers allow proton translocation across the mitochondrial inner membrane, causing loss of MMP (Figure 7A). The presence of a positively charged side chain and the asymmetrical distribution of the MMP-retaining uncouplers result in an electrogradient that compensates for the loss of MMP due to proton translocation (Figure 7B). Figure 7C and Figure 7D show a general approach to create new MMP-retaining uncouplers by adding a positively charged side chain to a
-13-
SUBSTITUTE SHEET ( RULE 26 ) conventional uncoupler (Figure 7C), for example, by adding a tertiary amine- (or secondary amine-) containing side chain to obtain an MMP -retaining uncoupler (Figure 7D), where N is nitrogen; and X and Y are optimally substituted side chains. Figure 7E shows an example of an MMP -retaining uncoupler, showing a conventional uncoupling component in the frame.
[0034] Figures 8A shows the acute lethal dose leading to the death of 50% of test animals (Acute LD50 or LD50) and the minimum efficacious dose (MED) of the conventional uncoupler DNP (US EP A, 2, 4-dinitrophenol. https://www.epa.gov/sites/default/files/2016-09/documents/2-4-dinitrophenol.pdf, Figure 8A) and MMP -retaining uncouplers, Compound 64 (#64, Figure 8B) and Compound 25 (#25, Figure 8C). Compound 64 and Compound 25 exhibit drastically improved acute toxicity profiles over DNP. Acute toxicity tests were performed on C57B16 mice, male, 6 weeks old, n=6. Compounds 64 and 25 were prepared to a fine suspension in 0.5% CMC-Na / 1% Tween80 water solution. The tests were performed under fed condition, by oral gavage of 100-400 pL compound suspension according to the body weight to desired dosages (mg/kg). The mice were supplied with drinking water all through the testing time. Mouse behaviors were monitored every 15-30 minutes and LD50 were determined. MED was determined in diabetic and steatosis mouse models (Table 9).
[0035] Figures 9 shows the safety profile of Compound 64 (#64) compared to DNP. Figure 9A shows the NOAEL(no-observable-adverse-effect-level)/MED of DNP. https://www.atsdr.cdc.gov/ToxProfiles/tp64.pdf; (US EP A, 2, 4-dinitrophenol. https://www.epa.gov/sites/default/files/2016-09/documents/2-4-dinitrophenol.pdf; and US CDC). Figure 9B shows the MED, intermediate NOAEL and NOAEL/MED of Compound 64 calculated by oral dosage. Figure 9C shows the MED, intermediate NOAEL and NOAEL/MED of Compound 64 over DNP calculated by Cmax (maximal blood concentration). Figure 9D shows the MED, intermediate NOAEL and NOAEL/MED of Compound 64 calculated byAUC (Area Under Curve). The Figures demonstrate that the MMP-retaining uncoupler, Compound 64 exhibits drastically improved short-term safety profiles over the conventional uncoupler DNP. A 10-day toxicology study in CD-I mice was performed to determine NOAEL of Compound 64, where animals (n=5) were administered once daily for 10 consecutive days. Clinical signs, body weights, food consumptions, rectal temperature, hematology, serum chemistry, plasma exposure, necropsies, tissue histopathology, were examined. MED was determined in diabetic and steatosis mouse models (Table 9)
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SUBSTITUTE SHEET ( RULE 26 ) [0036] Figure 10 show the safety profile of Compound 25 (#25) Figure 10A shows the MED, intermediate NOAEL and NOAEL/MED of Compound 25 calculated by oral dosage. Figure 10B shows the MED, intermediate NOAEL and NOAEL/MED of Compound 25 over DNP calculated by Cmax (maximal blood concentration). Figure 10C shows the MED, intermediate NOAEL and NOAEL/MED of Compound 25 calculated byAUC (Area Under Curve). The Figures demonstrate that the MMP -retaining uncoupler, Compound 25 exhibits drastically improved short-term safety profiles over the conventional uncoupler DNP. A 10-day toxicology study in CD-I mice was performed to determine NOAEL of Compound 25, where animals (n=5) were administered once daily for 10 consecutive days. Clinical signs, body weights, food consumptions, rectal temperature, hematology, serum chemistry, plasma exposure, necropsies, tissue histopathology, were examined. MED was determined in diabetic and steatosis mouse models (Table 9).
[0037] Figure 11 shows the efficacy of Compound 25 (#25) and Compound 64 (#64) in reducing blood glucose and glycated hemoglobin A1C in the db/db diabetic mouse model. Figure 11A is a graph of the blood glucose level of mice treated with vehicle and mice treated with Compound 25. Figure 1 IB is a graph of glycated hemoglobin A1C of mice treated with vehicle and mice treated with Compound 25. Figure 11C is a graph of the blood glucose level of mice treated with vehicle and mice treated with Compound 64. Figure 1 ID is a graph of glycated hemoglobin A1C of mice treated with vehicle and mice treated with Compound 64. BKS db/db mice were treated with or without 5mg/kg Compound 25 or Compound64 by daily oral gavage, for 3 weeks. Blood glucose and glycated hemoglobin A1C levels were measured. Statistical significance (P) was determined by the Student’s t- test. All error bars, s.d., ***p < 0.001. n=6 in each group. Vehicle, vehicle- treated control group.
[0038] Figure 12 shows the effects of Compound 25 on body weight (Figure 12A), blood glucose (Figure 12B), liver weight (Figure 12C), and plasma insulin levels (Figure D) in HFD-induced diabetic/fatty liver mouse model (n=6 in each group). Statistical significance (P) was determined by the Student’s t-test. All error bars, s.d., *p < 0.05, ***p < 0.001. n=6 in each group. #25: mice treated with Compound 25 (5mg/kg/day, daily gavage) for three weeks. Vehicle, vehicle-treated control group. Figure 12 A-B, Light bars, initial levels; dark bars, levels after 3 -week vehicle or drug treatment.
[0039] Figure 13 shows the effects of Compound 64 on blood glucose (Figure 13A) and plasma insulin levels (Figure 13B) in HFD-induced diabetic/fatty liver mouse model (n=6 in each group). Statistical significance (P) was determined by the Student’ s t-test.
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SUBSTITUTE SHEET ( RULE 26 ) All error bars, s.d., *p < 0.05, ***p < 0.001. n=6 in each group. #64: mice treated with Compound 64 (5mg/kg/day, daily gavage) for two weeks. Vehicle, vehicle-treated control group. Figure 13 A, Dark bars, initial levels; light bars, levels after 2-week vehicle or drug treatments.
[0040] Figure 14 shows the effects of Compound 25 on blood triglyceride (Figure 14A), total cholesterol (Figure 14B), and nori-HDI. cholesterol levels (Figure 14C) in high-fat diet induced diabetic/hepatic steatosis mice. Statistical significance (P) was determined by the Student’s t-test. All error bars, s.d., *p < 0.05, n=6 in each group. #25: treated with Compound 25 (5 mg/kg/day, daily gavage) for 3 weeks; vehicle, vehicle-treated control group.
[0041[ Figure 15 shows the effects of Compound 25 and Compound 64 on hepatic steatosis induced by high-fat diet. Representative liver histology images with H&E staining from mice as indicated: Figure 15A, normal: the liver section from a healthy C57/B16 mouse without HFD feeding; Figure 15B, HFD, the liver section from a mouse fed HFD; Figure 15C, HFD+#25, the liver sections from a mouse fed HFD following 3 weeks of Compound 25 treatment (P.O. dosing of 5 mg/kg, by daily gavage); and Figure 15D, HFD + #64, the liver sections from a mouse fed HFD following 3 weeks Compound 64 treatment (P.O. dosing of 5 mg/kg, by daily gavage). N=6 in each group.
[0042] Figure 16 shows the inhibitory effects of Compound 25 on hepatic stellate cell differentiation into myofibroblast-like cells. Figure 16A is a schematic representation of the experimental procedure (See procedure B5). Figure 16B is microscope images showing the cell morphology of LX-2 cells (human hepatic stellate cells) treated with vehicle, 10 ng/mL of TGFP and TGFP plus 7.5 ng/mL of Compound 25. TGFP treatment induces the differentiation of LX-2 cells into myofibroblast-like cells, which form the “ring”-shape morphology after TGFP treatment (TGFP, middle panel); Compound 25 (TGFp+#25, right panel) prevents TGFP-induced differentiation of LX-2 cells.
[0043] Figure 17 shows the inhibitory effects of Compound 64 on hepatic stellate cell differentiation into myofibroblast-like cells. Figure 17A. is microscope images showing cell morphology of LX-2 cells (human hepatic stellate cells) treated with vehicle, 10 ng/mL of TGFP and TGFP plus 1 ng/mL of Compound 64. TGF-P treatment induces the differentiation of LX-2 cells into myofibroblast-like cells, which form the “ring”-shape morphology after TGF-P treatment (TGF-P, middle panel); Compound64 (TGFp+#64, right panel) prevents TGFP-induced differentiation of LX-2 cells. Figure 17B is an immunoblotting analysis showing Compound 64 blocks activation of TGF-P signaling
-16-
SUBSTITUTE SHEET ( RULE 26 ) pathway as evidenced by losing Smad2/3 phosphorylation. LX-2 cells were treated with either vehicle (control), or TGF-0 alone, or TGF-0 plus indicated concentrations of Compound 64 for 6 hrs. Cells were then harvested, and immunoblotting analyses were performed with antibodies against p-Smad2/3 (phosphorylated Smad2/3), Smad2/3, or GAPDH, as indicated.
[0044] Figure 18 shows the efficacy of Compound 25 in reducing liver fibrosis determined by histology and molecular analysis. Figure 18A is microscope images of liver sections of CCL treated mice, subjected to H & E staining (top left panel); CCI4 plus Compound 25 (7.5 mg/kg/day) treated mice subjected to H & E staining (bottom left panel); CCL treated mice, subjected to Picrosirius Red staining(staining fibrotic collagen, top right panel) and CCL plus Compound 25 (7.5 mg/kg/day) treated mice subjected to Picrosirius Red staining (staining fibrotic collagen, bottom left panel. Figure 18B depicts fibrosis and lipofuscin scoring of vehicle (control), CCL alone, and CCL+ Compound 25 (CCL+25) treated mice. Each slice in a pie represents one mouse. The severity of liver fibrosis and abundance of lipofuscin in each mouse is presented with a different shade of color. Lipofuscin is an intracellular aggregate of highly oxidized proteins and lipids that cannot be digested. Lipofuscin mainly accumulate in lysosomes in aged cells and cells under pathological conditions. Figure 18C shows the immunoblotting analysis of collagen expression (Collal) in mice treated with vehicle, CCL alone, CCL+ Compound 25 (CCL+#25), with the fibrotic scores of each sample listed between the immunoblotting panels, as indicated. GAPDH serves as an internal control. n=7; duration of treatment, 6 weeks. Histological and molecular markers show Compound 25 reduces fibrosis in CCL treated animals.
[0045] Figure 19 is an immunoblotting analysis that shows the inhibitory effect of Compound 6464 on TGF-0 activation in T-cells. Figure 19A is an immunoblotting analysis of Human Jurkat cells treated with either vehicle alone (first lanes), TGF-0 alone (second lanes), TGF-0 plus Compound 64 at 0.5, 1.0 or 2.0 pM of compound (third through 5th lanes), as indicated, for 6 hours. Figure 19B is an immunoblotting analysis of mouse primary T-cells (B) were treated with either vehicle alone (first lanes), TGF-0 alone (second lanes), or TGF-0 plus varying concentrations of Compound 64 atl.O or 2.0 pM of compound (third and fourth lanes), as indicated, for 6 hours. Immunoblotting analyses were performed with antibodies against p-Smad2/3 (phosphorylated Smad2/3), Smad2/3, or GAPDH, as indicated.
-17-
SUBSTITUTE SHEET ( RULE 26 ) Loss of Smad2/3 phosphorylation indicates the efficacy of Compound 64 in blocking TGF-P activation in T-Cells.
[0046] Figure 20 shows Compound 64 is efficacious in combinatory therapy with PD-1 antibody in treating metastatic cancer in mice. Figure 20A is the experimental design (see Example B 12). Briefly, C57/B16 mice intrahepatically transplanted with MC38 cancer cells (day 0) were subject to various treatments starting on day 7: aPD-1 or isotype, mice were either treated with PD-1 antibody or its isotype antibody (control) by intraperitoneal (IP) injection on indicated days (PD-1 antibody has a half-life of over 1 week in mice); #64 or vehicle, mice were either treated with Compound #64 or vehicle by daily gavage. Figure 20B is a table depicting outcomes of the experiments described in Figure 20A and Example B12. aPD-1 +#64 represents mice treated with PD-1 antibody (IP) and #64 (daily gavage); aPD-1 represents mice treated with PD-1 antibody (IP) and vehicle (gavage); control, represents mice treated with isotype antibody (IP) and vehicle (gavage); n is number of mice in each group. Tumor-positive is the number of tumor- bearing mice in each group; tumor- free is thenumber of tumor-free mice in each group; tumor-free % is thepercentage of tumor- free animals in each group. Fisher Exact 2x2 test is theP value comparing each experimental group to control group using statistical analysis with Fisher Exact 2x2 test. P<0.05 is indicative of a statistically significant difference.
[0047] Figure 21 shows antiviral activity (EC50), cytotoxicity (TC50), and specificity index (SI) of Compounds 64, 25 and 57 against enveloped viruses. Each row represents results from experiments of the indicated compound on the host cells infected with or without the indicated virus. EC50 isthe compound concentration that reduces virus- induced cytopathic effects (CPE) by 50%. TC50 isthe compound concentration that leads to 50% of cell viability of uninfected cells; SI is the ratio between TC50 and EC50. The experiments were performed as follows. The host cells, either Vero 760 or MRC-5, as indicated, were seeded in 96-well flat-bottom tissue culture plates and allowed to adhere overnight. Following overnight incubation, the cells were either infected with virus (either SARS-CoV-2 or alpha coronavirus 229E) or uninfected, and diluted test compounds were added to each well. Following incubation at 37°C, 5% CO2 for three days or six days, cell viability was determined. Percent of CPE reduction of the virus-infected wells and the percent of cell viability of uninfected drug control wells, were measured to calculate and determine the EC50 and TC50 values. SI, was calculated accordingly.
-18-
SUBSTITUTE SHEET ( RULE 26 ) DETAILED DESCRIPTION
10048] The following factors make mitochondria an ideal target for treating a number of important diseases: (1) mitochondria are the ultimate site where lipid or glucose metabolites are consumed (oxidized); (2) mitochondria are critical in regulating the abundance of metabolic intermediates that become building blocks of biosynthesis essential for cell growth and proliferation of cancer cells, as well as for viral envelop production and assembly; (3) mitochondria are the major production site of ROS in neurons and many other cells (Figure 1).
[0049J Mitochondrial uncoupling is a unique way to modulate mitochondrial activity and functions. In essence, mitochondrial uncoupling is a process by which the activity of mitochondrial electron transport chain is de-coupled from ATP synthesis. Mechanistically, mitochondrial uncoupling is caused by the action of mitochondrial uncouplers that carry protons across the mitochondrial inner membrane into the mitochondrial matrix, independent of the ATP synthase (Terada, H. (1990) Environmental Health Perspectives 87, 213-218). The technical definition of mitochondrial uncouplers is an increase of oxygen consumption rate (OCR) by cells in the presence of an ATP synthase inhibitor such as oligomycin. As a result, mitochondrial uncouplers lead to futile mitochondrial oxidation and increase electron transport chain flux. Consequently, mitochondrial uncouplers could facilitate glucose or lipid catabolism in cells, reduce small molecule building block production output, and reduce electron stall in electron transport chain, hence reduce electron leakiness and mitochondrial ROS production. Through these actions, mitochondrial uncouplers represent an effective strategy for potential treatment of a number of important diseases.
|0050] Mitochondria and metabolic diseases. Metabolic diseases are a family of diseases characterized by symptoms of abnormal glucose and/or lipid metabolism, such as obesity, type 2 diabetes, alcoholic fatty liver disease, non-alcoholic fatty liver diseases, nonalcoholic steatohepatitis. These diseases are associated with age-, environmental-, or genetic- related decrease in mitochondrial functions such as reduced oxidative capacity. Importantly, these diseases also share a common causal factor, namely abnormal accumulation of intracellular lipid in cells of various tissues as well as insulin resistance in most cases. For example, obesity is characterized by excessive fat accumulation in cells of adipose tissue. Metabolic syndrome is characterized with insulin resistance in peripheral tissues, usually caused by ectopic fat accumulation in cells of liver, muscle, or adipose tissue. Type 2
-19-
SUBSTITUTE SHEET ( RULE 26 ) diabetes is characterized by insulin resistance usually caused by ectopic fat accumulation in cells of liver, muscle, or adipose tissue, and hyperglycemia caused by insulin resistance. Alcoholic fatty liver disease is characterized by ectopic lipid accumulation in liver cells and liver damage, liver inflammation, and fibrosis. The various stages of non-alcoholic liver fatty liver disease (orNAFLD), include hepatosteatosis, non-alcoholic steatohepatitis (NASH), cirrhosis, and NAFLD induced hepatocellular carcinoma (HCC), has a primary cause of ectopic lipid accumulation in liver cells which lead to liver damage, inflammation, and fibrosis. The various types of dyslipidemia are partly caused by ectopic accumulation of lipid in cells of liver, muscle, or heart, as a result of redistribution of lipid from adipose tissue to other tissues.
10051] Mitochondrial uncouplers, which reduce energy efficiency and boost futile lipid oxidation, would effectively reduce cellular accumulation of lipid. As ectopic intracellular accumulation of lipid in the liver and muscle, as well as excessive accumulation of lipid in adipose tissue are the fundamental cause of insulin resistance in various forms of metabolic diseases (Samuel V.T., et al., Lancet, 2010, 375:2267-77), applicant and others have demonstrated in animal models, that small molecule (chemical) mitochondrial uncouplers are efficacious in preventing and treating metabolic diseases (Tao, H., Zhang, Y., Zeng, X., Shulman, G. I., and Jin, S. ( 2014) Nature Medicine , 20, 1263-1269; Perry, R. J., Zhang, D., Zhang, X. M., Boyer, J. L., and Shulman, G. I. (2015) Science, 347(6227), 1253- 6), leading to: (1) reduction of lipid accumulation in various tissues, including adipose tissue, (2) reduction in insulin resistance, (3) reduction in blood glucose concentrations, and (4) improvement in glycemic control and slowdown in disease progression. Importantly, using mitochondrial uncouplers for treating metabolic diseases has a number of appealing features; for example, since they correct the cause of insulin resistance (ectopic lipid accumulation), such an approach may provide a cure for some metabolic diseases.
|0052] Cancer is a family of diseases characterized by uncontrolled growth and proliferation of cells of various tissue types, resulting from a combination of genetic mutations in oncogenes and tumor suppressor genes. It is well-accepted that one requirement of tumorigenesis is the alteration of cell metabolism. Cancer cells require not only energy but also the building blocks (metabolic intermediates) for biosynthesis of macromolecules such as DNA and RNA to support rapid cell growth and proliferation. Metabolism in cancer cells is changed in such a way it could support both the energy need and the need of the various metabolic intermediates (building blocks) for biosynthesis of macromolecules (Vander Heiden, M. G , Cantley, L. C., and Thompson, C. B. (2009) Science, 324(5930), 1029-33). As
-20-
SUBSTITUTE SHEET ( RULE 26 ) a result, most cancers exhibit a unique cellular metabolic pattern called the Warburg effect, or aerobic glycolysis, which prevents the complete oxidation of glucose or lipid and allows for production of glucose metabolites for biosynthesis of macromolecules (Vander Hei den, et al., 2009).
[0053] Mitochondrial uncoupling reduces energy efficiency thereby undermining the energy requirement of cancer cells. In addition, mitochondrial uncoupling promotes the complete mitochondrial oxidation of glucose and lipid, thereby diminishing the production of metabolic intermediates essential for biosynthesis of macromolecules required for cell proliferation. Moreover, mitochondrial uncoupling could lead to AMPK activation, a known event for inhibiting cell growth. Indeed, prior documents showed that mitochondrial uncouplers exhibit anti-cancer activities (U. S. Patent 10,227,315). Targeting cancer cells through mitochondrial uncoupling would deprive energy as well as biosynthetic metabolic intermediates that are absolutely essential for cancer cell growth and proliferation, which is proven to be an effective anti-cancer strategy (Alasadi, A. et al., (2018) Cell Death Dis., 9(2), 215)
[0054] Autoimmune diseases are conditions where the body’s immune system attacks their own healthy organs. The common autoimmune diseases include celiac disease, diabetes mellitus type 1, Graves' disease, inflammatory bowel disease, multiple sclerosis, psoriasis, rheumatoid arthritis, and systemic lupus erythematosus. To activate and sustain the autoimmune activity, the body’s own self-attaching immune cells need amplification (proliferation) which requires metabolic changes similar to the Warburg effect observed in cancer cells to provide sufficient building blocks for biosynthesis (Ganeshan, K., et al. (2014) Annual Review of Immunology, 32, 609-634). Therefore, mitochondrial uncoupling would potentially inhibit the activation and amplification of self-attacking immune cells.
[0055] Neurodegenerative diseases are a large group of disabling disorders of the nervous system such as Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and Alzheimer's disease, characterized by the relative selective death of neuronal subtypes. There is overwhelming evidence of impaired mitochondrial function as a causative factor in these diseases, with mitochondrial ROS production as one of the most important one (Elfawy, H. A., and Das, B. (2019) Life Sci., 218, 165-184.)
|0056] Mitochondrial uncouplers increase mitochondrial electron transport chain flux and decrease the electron stall in the complexes of the electron transport chain, thus they could effectively reduce mitochondrial ROS. Therefore, mitochondrial uncoupling is
-21-
SUBSTITUTE SHEET ( RULE 26 ) considered a powerful anti-oxidant strategy and could have therapeutic potential for treating neurodegenerative diseases.
|0057] A decline in mitochondrial function and capacity has been associated with normal aging and correlated with the development of a wide range of age-related diseases. Some proven ant-aging and pro-longevity approaches, such as calorie restriction, highly correlate with improvement in mitochondrial functions (Sun, N., et al. (2016) Mol. Cell, 61(5), 654-666). Modulation of mitochondrial functions by mitochondrial uncoupling has been proposed as anti-aging strategy (Caldeira da Silva, C. C., et al. (2008) Aging Cell, 7(4), 552-60).
[0058] Mitochondria are ancient bacteria that formed a symbiotic relationship with host cells. Bacterial plasma membrane contains electron transport chain and ATP synthase that are similar to those of mitochondria, therefore compounds that impact mitochondrial uncoupling could be useful inhibitors of bacterial growth and effective as antibiotics (US Patent 10,227,315).
[00591 Despite the appealing beneficial features of mitochondrial uncoupling, no
U. S. FDA approved drug is currently being used in humans with the mechanism of action of mitochondrial uncoupling for treating the above mentioned diseases in United States. There are major hurdles for developing mitochondrial uncouplers as therapeutics. Discovery of new synthetic mitochondrial uncouplers with better druggable features is critical for developing mitochondrial uncoupling therapeutics.
[0060] Benzamide mitochondrial uncouplers have been developed (International Patent Publication Number WO 2012/068274, International Patent Publication Number WO 2016/081599, U. S. Patent 10,227,315, and Tao et al., 2014) for potential therapeutic applications. One main limitation of the prior benzamide compounds are poor pharmacokinetic properties and low systemic exposure. For example, in order to overcome the poor pharmacokinetic properties of low systemic exposure and short half-life of the previously disclosed compounds, studies in animal models required the compound to be mixed with food, and high doses of the compounds were required to achieve efficacy (e.g. 1500 ppm niclosamide ethanolamine (NEN) in diet, equivalent to 150 mg/kg/day (Tao, et al. 2014, and WO 2012/068274); or 600-750 ppm of Compound 27 (in US10,227,315) in diet, equivalent of 60-75 mg/kg/day ). Neither the high doses required nor the need to mix the compound in food, is compatible with therapeutic development in humans.
[0061] Another limitation of the available mitochondrial uncouplers is safety concerns, because some mitochondrial uncouplers are known to have a narrow therapeutic
-22-
SUBSTITUTE SHEET ( RULE 26 ) index. For example, the most famous mitochondrial uncoupler, 2, 4- dinitrophenol , was previously used in humans but was withdrawn from the market due to its narrow therapeutic window. 2, 4- Dinitrophenol has a ratio of concentration (in cells)/dosage in vivo), between toxicity and efficacy, is merely a factor of about 3. Therefore, new properties that allow mitochondrial uncouplers that improve the safety margin are considered critical for therapeutic development of mitochondrial uncouplers.
[0062] Various embodiments provide new benzamide mitochondrial uncouplers that effectively induce mitochondrial uncoupling (increase mitochondrial oxygen consumption in the presence of oligomycin) without significantly decreasing mitochondrial membrane potential (vide infra) over wide concentration ranges. These compounds are referred to herein as mitochondrial membrane potential-retaining (MMP -retaining) compounds (MMP-retaining).
[0063] These MMP-retaining compounds exhibit drastically improved safety profiles compared to the bench mark conventional mitochondrial uncoupler, DNP. These compounds exhibit a wider therapeutic index when used for treating metabolic diseases.
[0064] Various embodiments provide new benzamide mitochondrial uncouplers that show markedly improved pharmacokinetic properties, such as markedly increased systemic exposure (with some compounds having more than 100 fold increase in AUC than some previously disclosed druggable benzamide mitochondrial uncouplers with the highest reported systemic exposure).
[0065] Various embodiments describe the use of these compounds for the prevention and treatment of bacterial infections; dermatological disorders; viral infection; metabolic diseases or disorders , including, but not limited to, obesity, metabolic syndrome, type 2 diabetes, alcoholic fatty liver disease, non-alcoholic fatty liver diseases, dyslipidemia, and primary cancer of various tissue origins, and metastatic cancer.
|0066] Various embodiments provide the use of these compounds for treatment of disease symptoms such as hyperglycemia, insulin resistance, aberrant lipid accumulation, fibrosis, and aberrant TGF-P activation.
|0067] Various embodiments describe the use of these compounds for the prevention and treatment of the diseases alone or in combination with another agent.
-23-
SUBSTITUTE SHEET ( RULE 26 ) Compounds
[0068] In one embodiment, the present disclosure describes a compound of
Formula A: or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
[0069] The substituent R1000a of Formula A is selected from the group consisting of
-CH3, -CH2CH3, -Ci-C6 alkyl, -C3-C6 cycloalkyl, -OCH3, -CH2OCH3, -CH2OCH2OCH3,
C(O)N(CH2CH2OCH3)2. Each of substituents R5000A and R5000B, is independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH2)2OCH3; with the proviso that R5000A and R5000B are not both Ci-Ce alkyl. Alternatively R5000A and R5000B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl. Integer r’ is an integer selected the group consisting of 1, 2 and 3. Substituent R6000 is selected from the group consisting of a 5 to 6-membered heterocyclyl, pyridinyl and thiazolyl; and r is an integer selected from the group consisting of 0, 1, 2, and 3.
[0070] Substituent R1000c of Formula I is selected from the group consisting of chloro, fluoro, iodo, and bromo.
[0071] Each of substituents R4000b and R4000d of Formula A is independently selected from the group consisting of Y1000 and Z1000, provided that when R4000b is Y, R4000d is Z and when R4000b is Z, R4000d is Y. Substituent Y1000 is selected from the group consisting of
-24-
SUBSTITUTE SHEET ( RULE 26 ) chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-C6)alkyl, halo(Ci-C6)alkyl, -OCF3, - SO2(Ci-Ce)alkyl, cyano, and -CO2(Ci-C6)alkyl. Substituent Z1000 is selected from the group consisting of H,
-CH2OCH2CH3,-CH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3, -CH2O(CH2)2N(CH3)2, -CH2O(CH2)2NHSO2CH3, -(CH2)O(CH2)2NR2000AR2000B, -(CH2)SR3000, -CH2OCH2Ar', -OCH3 CH2NHC(O)CH2CH3, -CH2NHC(O)CH2OCH3, -CH2NHSO2CH3, -(CH2)t’NR7000AR7000B, and -(CH2)tR8000 provided that when Z1000 is H; R1000a is not Ci- G, alkyl, -C3-C6 cycloalkyl, -CH3 or -CH2CH3. Substituents R2000A and R2000B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclic ring optionally substituted with one or more methyl groups. Substituent R3000 is a 5 to 6-membered heterocyclic ring and s is an integer selected from the group consisting of 0, 1, 2 and 3. Substituent Ar1 is a 5-6 membered aryl or heteroaryl group optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl, halo, hydroxyl, and alkoxy. Each of R7000A andR7000B is independently selected from
Ci-Ce alkyl. Alternatively R7000A and R7000B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents independently selected from Ci-Ce alkyl and t’ is an integer selected from the group consisting of 1, 2, and 3. Substituent R8000 is selected from the group consisting of a
5 to 6-membered heterocyclic ring optionally substituted with methyl, and t is an integer selected from the group consisting of 0, 1, 2 and 3.
[0072] In some embodiments R4000d is selected from the group consisting of H, R4d as described in paragraph [0078] and R400d as described in paragraph [0149],
[0073] In some embodiments the compound of Formula A is a compound selected from the group consisting of a compound of Formula I, a compound of Formula II and a compound of Formula III as defined herein.
|0074] In some embodiments a compound of Formula A is selected from the group consisting of a compound as described in any of paragraphs [0089], [0120] and [0156], [0075] In one embodiment, the present disclosure describes a compound of Formula I:
-25-
SUBSTITUTE SHEET ( RULE 26 ) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
[00761 The substituent Rla of Formula I is selected from the group consisting of
-CH3, -CH2CH3, -0CH3, -CH2OCH3, -CH2OCH2OCH3, -CH2O(CH2)2OH, -
Each of substituents R5A andR5B, is independently selected from the group consisting of -Ci- Ch> alkyl; and -C1-C6 alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH2)2OCH3. Alternatively R5A and R5B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido, -SO2CH3,
-CF3, Ci-Ce alkyl, halo, and acyl. Integer m’ is an integer selected the group consisting of 1, 2 and 3. Substituent R6 is selected from the group consisting of a 5 to 6-membered heterocyclyl, pyridinyl and thiazolyl; and m is an integer selected from the group consisting of 0, 1, 2 and 3.
[0077] Substituent Rlc of Formula I is selected from the group consisting of chloro, fluoro, iodo, and bromo.
[0078] Each of substituents R4b and R4d of Formula I is independently selected from the group consisting of Y and Z, provided that when R4b is Y, R4d is Z and when R4b is Z, R4d is Y. Substituent Y is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 ,
-CHF2, fluoro(Ci-Ce)alkyl, halo(Ci-Ce)alkyl, -OCF3, -SO2(C1-C6)alkyl, cyano, and -CO2(Ci-C6)alkyl. Substituent Z is selected from the group consisting of -CH2OH, -CH2OCH2CH3, -CH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3, -CH2O(CH2)2NHCH3,
-26-
SUBSTITUTE SHEET ( RULE 26 ) -CH2O(CH2)2N(CH3)2, -CH2O(CH2)2NHSO2CH3, -(CH2)O(CH2)2NR2AR2B, -(CH2)nR3, -CFFOCFFAr, and OCH3. Substituents R2A and R2B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclic ring optionally substituted with one or more methyl groups. Substituent R3 is selected from the group consisting of a 5 to 6- membered heterocyclic ring and phenoxy; and n is an integer selected from the group consisting of 0 1, 2 and 3. Substituent Ar is a 5-6 membered aryl or heteroaryl group optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl, halo, hydroxy, and alkoxy.
[0079] Further embodiments describe the compound according to Formula la: or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein each of Rla and R4d of Formula la is as described previously for Formula I.
[0080] In some embodiments, Rla of Formula I, or of Formula la or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from -CH2NR5AR5B; wherein each of R5A and R5B is independently selected from methyl; and Ci-Ce alkyl substituted with one or more groups selected from methoxy, and -O(CH2)2OCH3.
10081| In some embodiments, Rla of Formula I, or of Formula la or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NR5AR5B; wherein each of R5A and R5B is independently selected from Ci-Ce alkyl substituted with one or more methoxy.
[00821 In some embodiments, Rla of Formula I, or of Formula la or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NR5AR5B; wherein R5A and R5B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy,
-acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl. In some embodiments the 4-8-membered heterocyclyl is selected from the group consisting of azetindinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, morpholinyl,
-27-
SUBSTITUTE SHEET ( RULE 26 ) dioxothiomorphylinyl, and azabicyclo[3.2.1]octanyl; wherein the heterocyclyl is optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy,
-acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl. In some embodiments the 4-8-membered heterocyclyl is optionally substituted with one or more substituents independently selected from the group consisting of oxo, cyano, hydroxyl, methoxy,
-NHC(0)CH3, -C(0)NH2, -SO2CH3, -CF3, methyl, fluoro, and acetyl. In some embodiments the 4-8-membered heterocyclyl is optionally substituted with one or more methyl substituents. In some embodiments the 4 to 8-membered heterocyclyl is selected from the group consisting of azetindinyl; pyrrolidinyl; pyrrolidinyl substituted with one or more substituents selected from the group consisting of cyano, hydroxyl, methoxy, -NHC(0)CH3, - C(0)NH2, -SO2CH3, -CF3 and methyl; piperidinyl; piperidinyl substituted with one or more substituents selected from the group consisting of -CF3, and fluoro; piperazinyl substituted with one or more substituents selected from the group consisting of oxo, methyl, and acetyl; morpholinyl; morpholinyl substituted with one or more methyl groups; dioxothiomorphylinyl, and azabicyclo[3.2.1]octanyl.
[0083] In some embodiments, Rla of Formula I, or of Formula la or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NR5AR5B; wherein R5A and R5B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, -acylamino, carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl. In some embodiments the 6-membered heterocyclyl is selected from the group consisting piperidinyl optionally substituted with one or Ci-Ce alkyl substituents, and morpholinyl optionally substituted with one or Ci-Ce alkyl substituents.
[0084] In some embodiments, Rla of Formula I, or of Formula la or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NR5AR5B; wherein -CH2NR5AR5B is selected from the group consisting of
SUBSTITUTE SHEET ( RULE 26 )
[0085] In some embodiments, Rla of Formula I, or of Formula la or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is -(CH2)mR6; wherein R6 is selected from the group consisting of tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, thiazolyl and piperidinyl and m is as previously described. In some embodiments m is 0. In some embodiments m is 1. In some embodiments -(CH2)m R6 is selected from the group
|0086] In some embodiments, Rla of Formula I, or of Formula la or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH3, -CH2CH3, -OCH3, -CH2OCH3, -CH2OCH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3, CH2)NH(CH2)2OCH3, -CH2NHC(O)CH3, -(CH2)2NHCO2CH3,
SUBSTITUTE SHEET ( RULE 26 )
[0087] In some embodiments, Rla of Formula I, or of Formula la or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH3, -CH2CH3 and -CH2NR5AR5B wherein R5A and R5B are as previously described in any embodiment described herein. In some embodiments, each of R5A and R5B is independently selected from Ci-Ce alkyl substituted with one or more methoxy; alternatively, R5A and R5B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl.
[0088] In some embodiments, Rla of Formula I, or of Formula la or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group
SUBSTITUTE SHEET ( RULE 26 )
[0089] In some embodiments, Rlc of Formula I, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is chloro.
[0090] Some embodiments describe a compound of Formula I, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein R4b is Y and R4dis Z.
|0091] Some embodiments describe a compound of Formula I, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein R4dis Y and R4b is Z.
[00921 In some embodiments, Y of Formula I, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is -CF3.
[0093] In some embodiments, Z of Formula I, or R4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is -(CH2)O(CH2)2NR2AR2B wherein R2A and R2B are as previously described herein. In some embodiments R2A and R2B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with a methyl group. In some embodiments R2A and R2B together with the nitrogen to which they are attached, form a heterocyclyl selected from piperazinyl or a 4- methyl piperazinyl.
[00941 In some embodiments, Z of Formula I, or R4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is -(CH2)nR3, wherein R3 is as previously described herein. In some embodiments R3 is a 5-membered heterocyclic ring. In some embodiments R3 is selected from the group consisting of tetrahydrofuranyl, and phenoxy. In some embodiments R3 is tetrahydrofuranyl. In some embodiments R3 is selected from the group consisting of and phenoxy. In some embodiments n is 0. In some embodiments n is 1. In some embodiments -(CH2)nR3 is selected from the group consisting
SUBSTITUTE SHEET ( RULE 26 ) [00951 In some embodiments, Z of Formula I, or R4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is -CH2OCH2Ar, wherein Ar is as previously described herein. In some embodiments Ar is a 5 to 6-membered aryl or heteroaryl optionally substituted with one or more substituents independently selected from the group consisting of methyl, fluoro, chloro, hydroxy, and methoxy. In some embodiments Ar is selected from the group consisting of thiazolyl, phenyl, and phenyl substituted with one or more groups independently selected from methyl, fluoro, chloro, hydroxy, and methoxy.
In some embodiments Ar is selected from the group consisting of, phenyl,
[0096] In some embodiments, Z of Formula I, or R4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2OH, -CH2OCH2CH3, -CH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3, CH2O(CH2)2NHCH3,-CH2O(CH2)2N(CH3)2, -CH2O(CH2)2NHSO2CH3, -(CH2)O(CH2)2 NR2AR2B, -(CH2)nR3, -CH2OCH2Ar and OCH3; wherein R2A R2B, R3, n and Ar are as described in any embodiment herein.
[0097] In some embodiments, Z of Formula I, or R4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2OCH2CH -CH2O(CH2)2N(CH3)2, -CH2 -CFLOCJTAr and OCH3; wherein R2A R2B, R3, n and Ar are as described in any embodiment herein.
[0098] In some embodiments, Z of Formula I, or R4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2OCH2CH3, -CH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3, -CH2O(CH2)2N(CH3)2, -CH2O(CH2)2NHSO2CH3, -(CH2)O(CH2)2NR2A R2B, -(CH2)nR3, -CFLOCJTAr and OCH3; wherein R2A and R2B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of piperazinyl and 4-methyl piperazinyl; R3 is tetrahydrofuranyl; and Ar is a 5 to 6-membered aryl or heteroaryl group
-32-
SUBSTITUTE SHEET ( RULE 26 ) optionally substituted with one or more substituents independently selected from the group consisting of methyl, fluoro, chloro, hydroxy, and methoxy.
|0099] In some embodiments Z of Formula I, or R4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2OH,-CH2OCH2CH3, -CH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3,
[0100} In some embodiments Z of Formula I, or R4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group
[0101 ] In some embodiments Z of Formula I or R4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2OH, -CH2OCH2CH3, -CH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3,
SUBSTITUTE SHEET ( RULE 26 ) f Formula I is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-Ce)alkyl, halo(Ci-Ce)alkyl, -OCF3, -SO2(Ci- Ce)alkyl, cyano, and -CO2(Ci-Ce)alkyl.
[0102] In some embodiments Z of Formula I or R4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2OH, -CH2OCH2CH3, -CH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3,
[0103] In some embodiments Z of Formula I or R4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group he group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-C6)alkyl, halo(Ci- C6)alkyl, -OCF3, -SO2(Ci-C6)alkyl, cyano, and -CO2(Ci-C6)alkyl.
[0104] In some embodiments Z of Formula I or R4d of Formula la, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting -CH2OCH2CH3, -CH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3,
SUBSTITUTE SHEET ( RULE 26 )
10105] Some embodiments describe a compound of Formula I wherein:
Rlais selected from the group consisting of -CH3, -CH2CH3 and -CH2NR5AR5B; each of R5A and R5B is independently selected from Ci-Ce alkyl substituted with one or more groups selected from methoxy; alternatively, R5A and R5B together with the nitrogen to which they are attached, form a 4-8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl;
Rlc is chloro; each of R4b and R4d is independently selected from the group consisting of Y and Z, provided that when R4b is Y, R4d is Z and when R4b is Z, R4d is Y;
Y is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-C6)alkyl, halo(Ci-C6)alkyl, -OCF3, -SO2(Ci-C6)alkyl, cyano, and -CO2(Ci- Ce)alkyl;
Z is selected from the group consisting of -CH2OCH2CH3,-CH2OCH3, - CH2O(CH2)2OH, -CH2O(CH2)2OCH3, -CH2O(CH2)2N(CH3)2J -CH2O(CH2)2NHSO2CH3, - (CH2)O(CH2)2 NR2A R2B, -(CH2)nR3, -CH2OCH2Ar and -OCH3; wherein R2A and R2B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of piperazinyl and 4-methyl piperazinyl; R3 is tetrahydrofuranyl; and Ar is a 5 to 6-membered aryl or heteroaryl group optionally substituted with one or more substituents independently selected from the group consisting of methyl, fluoro, chloro, hydroxy, and methoxy.
(0106] Some embodiments describe a compound of Formula I or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein:
SUBSTITUTE SHEET ( RULE 26 ) Rlais selected from the group consisting of -CH3, -CH2CH3 and -CH2NR5AR5B; wherein each of R5A and R5B is independently selected from Ci-Ce alkyl substituted with one or more groups selected from methoxy; alternatively, R5A and R5B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl;
Rlc is chloro; each of R4b and R4d is independently selected from the group consisting of Y and Z, provided that when R4b is Y, R4d is Z and when R4b is Z, R4d is Y; Y is CF3; and Z is selected from the group consisting of -CH2OCH2CH3, -CH2OCH3, -CFLCXCFb^OH, - CH2O(CH2)2OCH3, -CH2O(CH2)2N(CH3)2, -CH2O(CH2)2NHSO2CH3, -(CH2)O(CH2)2NR2A R2B, -(CH2)nR3, -CFLOCFLAr and -OCH3; wherein R2A and R2B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of piperazinyl and 4-methyl piperazinyl; R3 is tetrahydrofuranyl; and Ar is a 5 to 6-membered aryl or heteroaryl group optionally substituted with one or more substituents independently selected from the group consisting of methyl, fluoro, chloro, hydroxy, and methoxy.
[0107] Some embodiments describe a compound of Formula I or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein:
Rlais selected from the group consisting of -CH3, -CH2CH3, -
Rlc is chloro; each of R4b and R4d is independently selected from the group consisting of Y and Z, provided that when R4b is Y, R4d is Z and when R4b is Z, R4d is Y; Y is CF3; and Z is selected from the group consisting of -CH2OCH2CH3, -CH2OCH3, -CH2O(CH2)2OH, -
SUBSTITUTE SHEET ( RULE 26 )
[01081 Some embodiments describe a compound of Formula I or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein:
Rlcis chloro;
R4b is CF3; and
R4d is selected from the group consisting of -CH2OCH2CHa, -CH2OCH3,
[0109] Some embodiments describe a compound of Formula la or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein Rlais selected from the H, -
-37-
SUBSTITUTE SHEET ( RULE 26 ) CH2OCH2CH3, -CH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3, -CH2O(CH2)2NHCH3, -
(0110] Some embodiments describe a compound of Formula la or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein Rlais selected from the group consisting of
(0111] Some embodiments describe a compound selected from the group consisting of:
SUBSTITUTE SHEET ( RULE 26 )
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26) or a pharmaceutically acceptable salt, solvate, or prodrug thereof
|0H2] Some embodiments describe a compound selected from the group consisting of:
SUBSTITUTE SHEET ( RULE 26 )
SUBSTITUTE SHEET (RULE 26) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
[0113] In one embodiment the compound is or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
[0114] In one embodiment, the present disclosure describes a compound of
Formula II:
-49-
SUBSTITUTE SHEET ( RULE 26 ) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
[01151 The substituent R10a of Formula I is selected from the group consisting of
-(CH2)O’NR50AR50B , -(CH2)OR60, and C(O)N(CH2CH2OCH3)2. Each of substituents R50A and R50B, is independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH2)2OCH3, with the proviso that R50Aand R50B are not both Ci-Ce alkyl; alternatively R50A and R50B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl. Integer o’ is an integer selected the group consisting of 1, 2 and 3.
Substituent R60 is selected from the group consisting 5 to 6-membered heterocyclyl, pyridinyl and thiazolyl; and o is an integer selected from the group consisting of 0 1, 2 and 3.
Substituent R10c of Formula I is selected from the group consisting of chloro, fluoro, iodo, and bromo.
[01171 One of substituent R40b and R40d is H and the other substituent is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-C6)alkyl, halo(Ci-C6)alkyl, -OCF3, -SO2(Ci-C6)alkyl, cyano, and -CO2(Ci-C6)alkyl.
[01181 Further embodiments describe the compound according to Formula Ila:
-50-
SUBSTITUTE SHEET ( RULE 26 ) or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein each of R10a, and R40d of Formula Ila is as described previously for Formula II.
[0119| In some embodiments, R10a of Formula II, or of Formula Ila or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from - CH2NR50AR50B; wherein each of R50Aand R50B is independently selected from methyl; and Ci-Ce alkyl substituted with one or more groups selected from methoxy, and -O(CH2)2OCH3.
[0120] In some embodiments, R10a of Formula II, or of Formula Ila or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NR50AR50B; wherein R50A and R50B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy,
-acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl. In some embodiments the 4-8-membered heterocyclyl is selected from the group consisting of azetindinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, morpholinyl, dioxothiomorphylinyl, and azabicyclo[3.2.1]octanyl; wherein the heterocyclyl is optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy,
-acylamino, carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl. In some embodiments the 4-8-membered heterocyclyl is selected from the group consisting of azetindinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, morpholinyl, and dioxothiomorphylinyl; wherein the heterocyclyl is optionally substituted with one or more substituents independently selected from the group consisting of oxo, cyano, hydroxyl, methoxy, -NHC(O)CH3, -C(O)NH2, -SO2CH3,
-CF3, methyl, fluoro, and acetyl. In some embodiments the 4 to 8-membered heterocyclyl is selected from the group consisting of azetindinyl; pyrrolidinyl; pyrrolidinyl substituted with one or more substituents selected from the group consisting of cyano, hydroxyl, methoxy, - NHC(O)CH3, -C(O)NH2, -SO2CH3, -CF3 and methyl; piperidinyl; piperidinyl substituted with one or more substituents selected from the group consisting of -CF3, and fluoro;
SUBSTITUTE SHEET ( RULE 26 ) piperazinyl substituted with one or more substituents selected from the group consisting of oxo, methyl, and acetyl; morpholinyl; morpholinyl substituted with one or more methyl groups; dioxothiomorphylinyl, and azabicyclo[3.2.1]octanyl. In some embodiments the 4 to 8-membered heterocyclyl is selected from the group consisting of azetindinyl; pyrrolidinyl; pyrrolidinyl substituted with one or more substituents selected from the group consisting of cyano, hydroxyl, methoxy, -NHC(0)CH3, -C(0)NH2, -SO2CH3, -CF3 and methyl; piperidinyl; piperidinyl substituted with one or more substituents selected from the group consisting of -CF3, and fluoro; piperazinyl substituted with one or more substituents selected from the group consisting of oxo, methyl, and acetyl; morpholinyl; morpholinyl substituted with one or more methyl groups; dioxothiomorphylinyl.
[0121 ] In some embodiments, R10a of Formula II, or of Formula Ila or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NR50AR50B; wherein R50A and R50B together with the nitrogen to which they are attached, form a 4-7 membered heterocyclyl selected from the group consisting of azetindinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, morpholinyl, and dioxothiomorphylinyl; wherein the 4-7 membered heterocyclyl is optionally substituted with one or more substituents independently selected from the group consisting of oxo, cyano, hydroxyl, methoxy, -NHC(0)CH3, -C(0)NH2, -SO2CH3, -CF3, methyl, fluoro, and acetyl.
[0122 | In some embodiments, R10a of Formula II, or of Formula Ila or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NR50AR50B; wherein -CH2NR50AR50B is selected from the group consisting
SUBSTITUTE SHEET ( RULE 26 )
[0123] In some embodiments, R10a of Formula II, or of Formula Ila or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NR50AR50B; wherein -CH2NR50AR50B is selected from the group consisting
[0124] In some embodiments, R10a of Formula II, or of Formula Ila or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is -(CH2)OR60; wherein R60 is selected from the group consisting of tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, thiazolyl, and pyridinyl and o is as previously described. In some embodiments o is 0. In some embodiments o is 1. In some embodiments R60 is selected from the group consisting of tetrahydrofuranyl, thiazolyl, and pyridinyl and o is 0. In some embodiments -(CH2)o R60 is
SUBSTITUTE SHEET ( RULE 26 ) selected from the group consisting
In some embodiments -(CH2)o R60 is selected from the group consisting of
[0125] In some embodiments R10a of Formula II, or of Formula Ila or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -OCH3, -CH2OCH3, -CH2OCH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3, C(O)N(CH2CH2OCH3)2, wherein R50A R50B , and R60 are as previously defined by any embodiment herein.
|0126] In some embodiments, R10a of Formula II, or of Formula Ila or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is -OCH3, -CH2OCH3,
SUBSTITUTE SHEET ( RULE 26 )
[0127] In some embodiments, R10a of Formula II, or of Formula Ila or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is -OCH3, -CH2OCH3,
SUBSTITUTE SHEET ( RULE 26 )
C(O)N(CH2CH2OCH3)2.
[0128] In some embodiments, R10c of Formula II, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is chloro, fluoro, and iodo.
[0129] In some embodiments, R10c of Formula II, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is chloro.
[0130] In some embodiments one of substituent R40b and R40d is H and the other substituent is selected from the group consisting of fluoro, -CF3 , -CHF2, and -OCF3.
[0131] In some embodiments, R40b is H and R40d is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-C6)alkyl, halo(Ci-Ce)alkyl, -OCF3,
-SO2(Ci-C6)alkyl, cyano, and -CO2(Ci-C6)alkyl.
[0132] In some embodiments, R40d is H and R40b is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-Ce)alkyl, halo(Ci-Ce)alkyl, -OCF3,
-SO2(Ci-C6)alkyl, cyano, and -CO2(Ci-C6)alkyl.
[0133] In some embodiments R40d is H and R40b is selected from the group consisting of fluoro, -CF3, -CHF2, and -OCF3.
[0134] In some embodiments R40d is H and R40b is -CF3.
[0135| Some embodiments describe a compound of Formula II or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein
R10a is selected from the group consisting of -OCH3, -CH2OCH3, -CH2OCH2OCH3, each of R50AandR50B is independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH2)2OCH3; alternatively R50A and R50B together with the nitrogen to which they are attached, form a 4 to 7-membered heterocyclyl optionally substituted with one or more
SUBSTITUTE SHEET ( RULE 26 ) substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl;
R60 is selected from the group consisting of 5 to 6-membered heterocyclyl, pyridinyl and thiazolyl; o is 0;
R10c is selected from the group consisting of chloro, fluoro, iodo; and one of R40b and R40d is H and the other of R40b and R40d is selected from the group consisting of fluoro, -CF , -CHF2, -OCF3; with the proviso that R50A and R50B are not both Ci-Ce alkyl.
|0136] Some embodiments describe a compound of Formula II or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein
R10a is selected from the group consisting of -OCH3, -CH2OCH3, -CH2OCH2OCH3, each of R50A and R50B is independently selected from methyl; and Ci-Ce alkyl substituted with one or more groups selected from methoxy, and -O(CH2)2OCH3; alternatively R50A and R50B together with the nitrogen to which they are attached, form a 4-7- membered heterocyclyl selected from the group consisting of azetindinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, morpholinyl, and dioxothiomorphylinyl; wherein the heterocyclyl is optionally substituted with one or more substituents independently selected from the group consisting of oxo, cyano, hydroxyl, methoxy, -NHC(O)CH , -C(O)NH2, - SO2CH3, -CF3, methyl, fluoro, and acetyl;
R60 is selected from the group consisting of tetrahydrofuranyl, thiazolyl, and pyridinyl o is 0;
R10c is selected from the group consisting of chloro, fluoro, iodo; and
R40d is H and R40b is selected from the group consisting of fluoro, -CF3, -CHF2, and -OCF3;
-57-
SUBSTITUTE SHEET ( RULE 26 ) with the proviso that R50Aand R50B are not both methyl.
[0137| Some embodiments describe a compound of Formula II or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein
C(O)N(CH2CH2OCH3)2;
R10c is selected from the group consisting of chloro, fluoro, iodo; and
R40d is H and R40b is selected from the group consisting of fluoro, -CF3, -CHF2, and -OCF3.
-58-
SUBSTITUTE SHEET ( RULE 26 ) [O138| Some embodiments describe a compound of Formula II or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein
R10c is selected from the group consisting of chloro, fluoro, iodo; and
R40d is H and R40b is selected from the group consisting of fluoro, -CF3, -CHF2, and -OCF3.
[0139 j Some embodiments describe a compound of Formula II or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein R10a is selected from the group consisting of -OCH3, -CH2OCH3, -CH2OCH2OCH3, -CH2O(CH2)2OH, - CH2O(CH2)2OCH3,
-59-
SUBSTITUTE SHEET ( RULE 26 )
R10c is chloro; and
R40d is H and R40b is -CF3.
|0140] Some embodiments describe a compound of Formula Ila or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein
R10a is selected from the group consisting of -OCH3, -CH2OCH3, -CH2OCH2OCH3,
SUBSTITUTE SHEET ( RULE 26 )
C(O)N(CH2CH2OCH3)2.
[0141] Some embodiments describe a compound of Formula Ila or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein
SUBSTITUTE SHEET ( RULE 26 )
[0142] Some embodiments describe a compound selected from the group consisting of:
SUBSTITUTE SHEET ( RULE 26 )
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
|0143] Some embodiments describe a compound selected from the group consisting of:
SUBSTITUTE SHEET ( RULE 26 )
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
|0144] One embodiment describes a compound that is
SUBSTITUTE SHEET ( RULE 26 ) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
[01451 In one embodiment, the present disclosure describes a compound of
Formula III: Ill or a pharmaceutically acceptable salt, solvate, or prodrug thereof
[0146] The substituent R100a of Formula III is selected from the group consisting of
-(CH2)p’NR500AR500B , -(CH2)pR600, and C(O)N(CH2CH2OCH3)2. Each of substituents R5A andR5B, is independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH2)2OCH3.
Alternatively R500A and R500B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido,
-SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl. Integer p’ is an integer selected the group consisting of 1, 2 and 3. Substituent R600 is selected from the group consisting of 5 to 6-
-75-
SUBSTITUTE SHEET ( RULE 26 ) membered heterocyclyl, pyridinyl and thiazolyl; and p is an integer selected from the group consisting of 0, 1, 2, and 3.
[0147| Substituent R100c of Formula III is selected from the group consisting of chloro, fluoro, iodo, and bromo.
10148] Each of substituents R400b and R400d of Formula III is independently selected from the group consisting of Y1 and Z1, provided that when R400b is Y1, R400d is Z1 and when R400b is Z1, R400d is Y1. Substituent Y1 is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-C6)alkyl, halo(Ci-C6)alkyl, -OCF3, - SO2(Ci-Cs)alkyl, cyano, and -CO2(Ci-Ce)alkyl. Substituent Z1 is selected from the group consisting of -CH2NHC(O)CH2CH3, -CH2NHC(O)CH2OCH3, -CH2NHSO2CH3, -(CH2)q’NR7AR7B, and -(CH2)qR8. Each ofR7AandR7B is independently selected from Ci-Ce alkyl. Alternatively R7A and R7B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more independently selected Ci-Ce alkyl. Integer q’ is an integer selected from the group consisting of 1, 2, and 3. Substituent R8 is selected from the group consisting of a 5 to 6-membered heterocyclic ring optionally substituted with methyl, and q’ is an integer selected from the group consisting of 0, 1, 2 and 3.
[0149] Further embodiments describe the compound according to Formula Illa: TTT Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein each of R100a, and R4°0d of Formula Illa is as described previously for Formula III.
[01501 In some embodiments, R100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from - CH2NR500AR500B; wherein each of R500A and R500B is independently selected from is independently selected from methyl; and Ci-Ce alkyl substituted with one or more groups selected from methoxy, and -O(CH2)2OCH3.
-76-
SUBSTITUTE SHEET ( RULE 26 ) [O151 | In some embodiments, R100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NR500AR500B; wherein each of R500A and R500B is independently selected from Ci-C6 alkyl substituted with one or more methoxy.
[0152] In some embodiments, R100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NR500AR500B; wherein R500A and R500B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, -acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl. In some embodiments the 4-8-membered heterocyclyl is selected from the group consisting of azetindinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, morpholinyl, dioxothiomorphylinyl, and azabicyclo[3.2.1]octanyl; wherein the heterocyclyl is optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, -acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl. In some embodiments the 4-8-membered heterocyclyl is optionally substituted with one or more substituents independently selected from the group consisting of oxo, cyano, hydroxyl, methoxy, -NHC(O)CH3, -C(O)NH2, -SO2CH3, -CF3, methyl, fluoro, and acetyl. In some embodiments the 4-8-membered heterocyclyl is optionally substituted with one or more methyl substituents. In some embodiments the 4 to 8-membered heterocyclyl is selected from the group consisting of azetindinyl; pyrrolidinyl; pyrrolidinyl substituted with one or more substituents selected from the group consisting of cyano, hydroxyl, methoxy, -NHC(O)CH3, - C(O)NH2, -SO2CH3, -CF3 and methyl; piperidinyl; piperidinyl substituted with one or more substituents selected from the group consisting of -CF3, and fluoro; piperazinyl substituted with one or more substituents selected from the group consisting of oxo, methyl, and acetyl; morpholinyl; morpholinyl substituted with one or more methyl groups; dioxothiomorphylinyl, and azabicyclo[3.2.1]octanyl.
10153] In some embodiments, R100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NR500AR500B; wherein R500A and R500B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with one or more
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SUBSTITUTE SHEET ( RULE 26 ) substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy,
-acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl In some embodiments the 6-membered heterocyclyl is selected from the group consisting piperazinyl optionally substituted with one or Ci-Ce alkyl substituents, and morpholinyl optionally substituted with one or Ci-Ce alkyl substituents.
[0154] In some embodiments, R100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NR500AR500B; wherein -CH2NR500AR500B is selected from the group embodiments -CH2NR500AR500B is selected from the group consisting of
|0155] In some embodiments, R100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is -(CH2)PR600; wherein R600 is selected from the group consisting of tetrahydrofuranyl, tetrahydropyranyl, piperidinyl,
SUBSTITUTE SHEET ( RULE 26 ) thiazolyl and piperidinyl and m is as previously described. In some embodiments p is 0. In some embodiments p is 1. In some embodiments -(CH2)PR600 is selected from the group
[O156| In some embodiments, R100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH3, -OCH3, -CH2OCH3, -CH2OCH2OCH3, -CH2O(CH2)2OH, -
SUBSTITUTE SHEET ( RULE 26 ) [0157] In some embodiments, R100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH3, and -CH2NR500AR500B wherein R500A and R500B are as previously described in any embodiment described herein. In some embodiments, each of R500A and R5°°B JS indepencientiy selected from Ci-Ce alkyl substituted with one or more methoxy; alternatively, R500A and R500B together with the nitrogen to which they are attached, form a 6- membered heterocyclyl optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl.
[0158] In some embodiments, R100a of Formula III, or of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group
[0159] In some embodiments, R100c of Formula III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is chloro.
[0160| Some embodiments describe a compound of Formula III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein R400b is Y1 and R400dis Z1.
[0161] Some embodiments describe a compound of Formula III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein R400d is Y1 and R400b is Z1.
[0162] In some embodiments, Y'of Formula III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is -CF3.
[0163] In some embodiments, Z'of Formula III, or R400d of Formula Illa, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is -CH2NR7AR7B wherein R7A and R7B are as previously described herein. In some embodiments each of R7A and R7B is methyl. In some embodiments R7A and R7B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with a methyl group. In some embodiments R7A and R7B together with the nitrogen to which they are attached, form a heterocyclyl
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SUBSTITUTE SHEET ( RULE 26 ) selected from the group consisting of piperazinyl optionally substituted with one or more Ci- Cc> alkyl and morpholinyl optionally substituted with one or more Ci-Ce alkyl. In some embodiments R7A and R7B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of, 4-methylpiperazinyl, and morpholinyl. In some embodiments
-CH2NR7AR7B is selected from the group consisting
[0164] In some embodiments, Z1 of Formula III, or R400d of Formula Illa, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is -(CH2)qR8, wherein R8 is as previously described herein. In some embodiments R8 is a 6-membered heterocyclic ring optionally substituted with methyl. In some embodiments R8 is 4-methylpiperidinyl. In some embodiments q is 0. In some embodiments q is 1. In some embodiments -(CH2)qR8 is
[01 5] In some embodiments, Z1 of Formula I, or R400d of Formula Illa, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NHC(O)CH2CH3, -CH2NHC(O)CH2OCH3, -CH2NHSO2CH3, - CH2NR7AR7B, and -(CH2)qR8, wherein R7A, R7B, R8, and p are as described in any embodiment herein.
[0166] In some embodiments, Z1 of Formula III, or R400d of Formula Illa, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NHC(O)CH2CH3, -CH2NHC(O)CH2OCH3, -CH2NHSO2CH3, - CH2NR7AR7B, and -(CH2)qR8 wherein R7A, R7B, R8, and p are as described in any embodiment herein.
[0167] In some embodiments, Z1 of Formula III, or R400d of Formula Illa, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NHC(O)CH2CH3, -CH2NHC(O)CH2OCH3, -CH2NHSO2CH3, - CH2NR7AR7B, and -(CH2)qR8, wherein each of R7A and R7B is methyl. Alternatively
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SUBSTITUTE SHEET ( RULE 26 ) embodiments R7A and R7B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of, 4-methylpiperazinyl, and morpholinyl; and R8 is 4-methylpiperidinyl.
|0168] In some embodiments , Z1 of Formula III, or R400d of Formula Illa, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of -CH2NHC(O)CH2CH3, -CH2NHC(O)CH2OCH3, -CH2NHSO2CH3, -
|0169] In some embodiments , Z1 of Formula III, or R400cl of Formula Illa, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group 3, -CH2NHSO2CH3, - Formula III is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-C6)alkyl, halo(Ci- C6)alkyl, -OCF3,
-SO2(Ci-Ce)alkyl, cyano, and -CO2(Ci-Ce)alkyl.
[0170 ] In some embodiments , Z1 of Formula III, or R400d of Formula Illa, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group 3, -CH2NHSO2CH3, - Formula III is -CF3 .
[0171] In some embodiments, R400d of Formula III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is selected from the group consisting of - , -CH2NHSO2CH3, -CH2N(CH3)2, of Formula III is -CF3.
|0172] Some embodiments describe a compound of Formula III or a pharmaceutically acceptable salt, solvate, or prodrug thereof
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SUBSTITUTE SHEET ( RULE 26 ) wherein:
R100ais selected from the group consisting of -CH3, and -CH2NR500AR500B; each of R500A and R500B is independently selected from Ci-Ce alkyl substituted with one or more methoxy; alternatively, R500A and R500B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl;
R1°°C jg chloro- each of R400b and R400d is independently selected from the group consisting of Y1 and Z1, provided that when R400b is Y1, R400d is Z1 and when R400b is Z1, R400d is Y1.
Y1 is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-C6)alkyl, halo(Ci-C6)alkyl, -OCF3, -SO2(Ci-C6)alkyl, cyano, and -CO2(Ci- C6)alkyl;
Z1 is selected from the group consisting of -CH2NHC(O)CH2OCH3, - CH2NHC(O)CH2CH3, -CH2NHSO2CH3, -CH2NR7AR7B, and -(CH2)qR8, wherein each of R7A and R7B is methyl; alternatively R7A and R7B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of, 4-methylpiperazinyl, and morpholinyl; and R8 is 4-methylpiperidinyl.
(0173| Some embodiments describe a compound of Formula III or a pharmaceutically acceptable salt, solvate, or prodrug thereof wherein:
R100ais selected from the group consisting of -CH3, and -CH2NR500AR500B; each of R500A and R500B is independently selected from Ci-Ce alkyl substituted with one or more methoxy; alternatively, R500A and R500B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl;
R1°°C js chloro- each of R400b and R400d is independently selected from the group consisting of Y1 and Z1, provided that when R400b is Y1, R400d is Z1 and when R400b is Z1, R400d is Y1.
-83-
SUBSTITUTE SHEET ( RULE 26 ) Y1 is -CF3;
Z1 is selected from the group consisting of-CH2NHC(O)CH2OCH3, - CH2NHC(O)CH2CH3, -CH2NHSO2CH3, -CH2NR7AR7B, and -(CH2)qR8, wherein each of R7A and R7B is methyl; alternatively R7A and R7B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of, 4-methylpiperazinyl, and morpholinyl; and R8 is 4-methylpiperidinyl
[0174] Some embodiments describe a compound of Formula III or a pharmaceutically acceptable salt, solvate, or prodrug thereof wherein:
R100ais selected from the group consisting of -CH3, -(CH2)N(CH2CH2OCH3)2,
R1°°C js chloro- each of R400b and R400d is independently selected from the group consisting of Y1 and Z1, provided that when R400b is Y1, R400d is Z1 and when R400b is Z1, R400d is Y1; Y1 is CF3; and Z1 is selected from the group consisting of -CH2NHC(O)CH2OCH3, -
[0175] Some embodiments describe a compound of Formula III or a pharmaceutically acceptable salt, solvate, or prodrug thereof wherein:
R100ais selected from the group consisting of -CH3, -(CH2)N(CH2CH2OCH3)2,
Rl°°c js chloj-Q-
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SUBSTITUTE SHEET ( RULE 26 ) R400b is CF3; and
[0176] Some embodiments describe a compound of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof wherein:
R100ais selected from the group consisting of -CH3, -(CH2)N(CH2CH2OCH3)2,
[0177] Some embodiments describe a compound of Formula Illa or a pharmaceutically acceptable salt, solvate, or prodrug thereof wherein:
|0178] Some embodiments describe a compound selected from the group consisting of:
SUBSTITUTE SHEET ( RULE 26 )
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
[O179| Some embodiments describe a compound selected from the group consisting of:
SUBSTITUTE SHEET ( RULE 26 )
[0180] Additional compounds that are mitochondrial uncouplers but may not necessarily have the unexpected properties of the other compounds disclosed herein include:
SUBSTITUTE SHEET ( RULE 26 )
SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
[0181] The compounds of embodiments described herein can possess one or more asymmetric carbon atoms and are thus capable of existing in the form of optical isomers as well as in the form of racemic or non-racemic mixtures thereof. The compounds can be utilized in embodiments described herein as a single isomer or as a mixture of stereochemical isomeric forms. Diastereoisomers, i.e., non-superimposable stereochemical isomers, can be separated by conventional means such as chromatography, distillation, crystallization or sublimation. The optical isomers can be obtained by resolution of the racemic mixtures according to conventional processes, for example by formation of diastereoisomeric salts by treatment with an optically active acid or base. Examples of appropriate acids include, without limitation, tartaric, diacetyltartaric, dibenzoyltartaric, ditoluoyltartaric and camphorsulfonic acid. The mixture of diastereomers can be separated by crystallization
SUBSTITUTE SHEET ( RULE 26 ) followed by liberation of the optically active bases from these salts. An alternative process for separation of optical isomers includes the use of a chiral chromatography column optimally chosen to maximize the separation of the enantiomers. Still another available method involves synthesis of covalent diastereoisomeric molecules by reacting compounds of the invention with an optically pure acid in an activated form or an optically pure isocyanate. The synthesized diastereoisomers can be separated by conventional means such as chromatography, distillation, crystallization or sublimation, and then hydrolyzed to obtain the enantiomerically pure compound. The optically active compounds of the invention can likewise be obtained by utilizing optically active starting materials. These isomers may be in the form of a free acid, a free base, an ester or a salt.
[0182] Compounds according to embodiments described herein may be in the form of pharmaceutically acceptable salts. A pharmaceutically acceptable salt of the compounds described herein includes acid addition salts and base addition salts. Pharmaceutically-acceptable salt embraces salts commonly used to form alkali metal salts and to form addition salts of free acids or free bases The nature of the salt is not critical, provided that it is pharmaceutically-acceptable. Suitable pharmaceutically-acceptable acid addition salts of the compounds described herein may be prepared from an inorganic acid or an organic acid. Examples of such inorganic acids include, without limitation, hydrochloric, hydrobromic, hydroiodic, nitric, carbonic, sulfuric and phosphoric acid. In some embodiments the salt is a hydrochloride salt. Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, arylaliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which include, without limitation, formic, acetic, propionic, succinic, glycolic, gluconic, maleic, embonic (pamoic), methanesulfonic, ethanesulfonic, 2- hydroxyethanesulfonic, pantothenic, benzenesulfonic, toluenesulfonic, sulfanilic, mesylic, cyclohexylaminosulfonic, stearic, algenic, P-hydroxybutyric, malonic, galactic, and galacturonic acid. Pharmaceutically-acceptable base addition salts for compounds described herein can be prepared from inorganic and organic bases. Salts derived from inorganic bases, include by way of example only, sodium, potassium, lithium, ammonium, calcium and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, such as alkyl amines, dialkyl amines, trialkyl amines, substituted alkyl amines, di(substituted alkyl) amines, tri(substituted alkyl) amines, alkenyl amines, dialkenyl amines, trialkenyl amines, substituted alkenyl amines, di (substituted alkenyl) amines, tri (substituted alkenyl) amines, cycloalkyl amines, di(cycloalkyl) amines, tri(cyclo alkyl) amines, substituted cycloalkyl amines, disubstituted cycloalkyl amine,
-93-
SUBSTITUTE SHEET ( RULE 26 ) trisubstituted cycloalkyl amines, cycloalkenyl amines, di(cycloalkenyl) amines, tri(cycloalkenyl) amines, substituted cycloalkenyl amines, disubstituted cycloalkenyl amine, trisubstituted cycloalkenyl amines, aryl amines, diaryl amines, triaryl amines, heteroaryl amines, diheteroaryl amines, triheteroaryl amines, heterocyclic amines, diheterocyclic amines, triheterocyclic amines, mixed di- and tri-amines where at least two of the substituents on the amine are different and are selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, heterocyclic, and the like. Also included are amines where the two or three substituents, together with the amino nitrogen, form a heterocyclic or heteroaryl group. Examples of suitable amines include, by way of example only, isopropylamine, trimethyl amine, diethyl amine, tri (iso-propyl) amine, tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-alkylglucamines, theobromine, purines, piperazine, piperidine, morpholine, N-ethylpiperidine, and the like. It should also be understood that other carboxylic acid derivatives would be useful in the preparation of pharmaceutically acceptable salts, for example, carboxylic acid amides, including carboxamides, lower alkyl carboxamides, dialkyl carboxamides, and the like.
[0183] Acceptable salts may be obtained using standard procedures well known in the art, for example by treating a sufficiently basic compound such as an amine with a suitable acid affording a physiologically acceptable anion. Alkali metal (for example, sodium, potassium or lithium) or alkaline earth metal (for example calcium) salts of organic (e g., carboxylic) acids can also be made.
[0184] Compounds of according to embodiments described herein may have prodrug forms. Any compound that will be converted in vivo to provide the bioactive agent is a prodrug within the scope and spirit of the invention. Various forms of prodrugs are well known in the art (see, for example, Medicinal Chemistry: Principles and Practice, F.D. King, ed., The Royal Society of Chemistry, Cambridge, UK, 1994; Hydrolysis in Drug and Prodrug Metabolism. Chemistry, Biochemistry and Enzymology, B. Testa, J. M. Mayer, VCHA and Wiley-VCH, Zurich, Switzerland, 2003; The Practice of Medicinal Chemistry, C. G. Wermuth, 2nd ed., Academic Press, San Diego, CA, 1999). Some prodrugs of the present invention include a compound according to any embodiment described herein in which the 2-
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SUBSTITUTE SHEET ( RULE 26 ) hydroxy of the benzamide is converted to a group such as, but not limited to,
10185] In some embodiments, a prodrug of a compound according to any embodiment described herein may take the form of a carbamate. For instance, the 2-hydroxy group of a benzamide according to any embodiment described may converted to a carbamate group, -OC(O)NR9R10 at the same position. Each of R9 and R10 is independently selected from the group consisting of hydrogen, and optionally substituted Ci-Ce-alkyl; alternatively R9 and R10 taken together with the nitrogen to which they are attached form an optionally substituted Cs- -heterocyclyl.
[0186| The invention also embraces isolated compounds. An isolated compound refers to a compound which represents at least 10%, preferably at least 20%, more preferably at least 50% and most preferably at least 80% of the compound present in the mixture.
[0187] In some embodiments of the invention, one or more hydrogen atoms is replaced by a deuterium. It is well established that deuteration of physiologically active compounds offer the advantage of retaining the pharmacological profile of their hydrogen counterparts while positively impacting their metabolic outcome. Selective replacement of one or more hydrogen with deuterium, in a compound of the present invention, could improve the safety, tolerability and efficacy of the compound when compared to its all hydrogen counterpart.
[0188] Methods for incorporation of deuterium into compounds is well established. Using metabolic studies establish in the art, the compound of the present invention can be tested to identify sites for selective placement of a deuterium isotope, wherein the isotope will not be metabolized. Moreover these studies identify sites of metabolism as the location where a deuterium atom would be placed.
10189] Some embodiments describe a pharmaceutical composition comprising: a compound according to an embodiment described herein, a pharmaceutically acceptable salt thereof, a solvate thereof, or a prodrug thereof; and a pharmaceutically acceptable carrier or diluent.
(0190] Compounds, or pharmaceutically acceptable salts thereof, can be formulated for oral, intravenous, intramuscular, subcutaneous or parenteral administration for the therapeutic or prophylactic treatment of diseases, disorders or infections described herein.
-95-
SUBSTITUTE SHEET ( RULE 26 ) For oral or parenteral administration, compounds of this invention can be mixed with conventional pharmaceutical carriers and excipients and used in the form of tablets, capsules, elixirs, suspensions, syrups, wafers and the like. The pharmaceutical compositions comprising a compound of this invention will contain from about 0.1 to about 99% by weight of the active compound, and more generally from about 10 to about 30%.
10191 | The pharmaceutical preparations disclosed herein are prepared in accordance with standard procedures and are administered at dosages that are selected to reduce, prevent or eliminate the infection (See, e. g., Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa. and Goodman and Gilman's. The Pharmaceutical Basis of Therapeutics, Pergamon Press, New York, N.Y., the contents of which are incorporated herein by reference, for a general description of the methods for administering various agents for human therapy). The pharmaceutical compositions of the invention can be delivered using controlled (e.g., capsules) or sustained release delivery systems (e.g., bioerodable matrices).
[0192| The pharmaceutically acceptable pharmaceutical compositions of the present invention comprise one or more compounds of the invention in association with one or more non-toxic, pharmaceutically acceptable carriers and/or diluents and/or adjuvants and/or excipients, collectively referred to herein as “carrier” materials, and if desired other active ingredients. The pharmaceutical compositions may contain common carriers and excipients, such as com starch or gelatin, lactose, sucrose, microcrystalline cellulose, kaolin, mannitol, dicalcium phosphate, sodium chloride and alginic acid. The pharmaceutical compositions may contain croscarmellose sodium, microcrystalline cellulose, com starch, sodium starch glycolate and alginic acid.
[0193| Tablet binders that can be included are acacia, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone (Povidone), hydroxypropyl methylcellulose, sucrose, starch and ethylcellulose.
[01941 Lubricants that can be used include magnesium stearate or other metallic stearates, stearic acid, silicone fluid, talc, waxes, oils and colloidal silica.
[01951 Flavoring agents such as peppermint, oil of wintergreen, cherry flavoring or the like can also be used. It may also be desirable to add a coloring agent to make the dosage form more aesthetic in appearance or to help identify the product.
[01961 For oral use, solid formulations such as tablets and capsules are particularly useful. Sustained release or enterically coated preparations may also be devised. For pediatric and geriatric applications, suspensions, syrups and chewable tablets are
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SUBSTITUTE SHEET ( RULE 26 ) especially suitable. For oral administration, the pharmaceutical compositions are in the form of, for example, a tablet, capsule, suspension or liquid. The pharmaceutical composition is preferably made in the form of a dosage unit containing a therapeutically effective amount of the active ingredient. Examples of such dosage units are tablets and capsules. For therapeutic purposes, the tablets and capsules which can contain, in addition to the active ingredient, conventional carriers such as binding agents, for example, acacia gum, gelatin, polyvinylpyrrolidone, sorbitol, or tragacanth; fillers, for example, calcium phosphate, glycine, lactose, maize-starch, sorbitol, or sucrose; lubricants, for example, magnesium stearate, polyethylene glycol, silica, or talc, disintegrants, for example, potato starch, flavoring or coloring agents, or acceptable wetting agents. Oral liquid preparations generally are in the form of aqueous or oily solutions, suspensions, emulsions, syrups or elixirs may contain conventional additives such as suspending agents, emulsifying agents, non-aqueous agents, preservatives, coloring agents and flavoring agents. Examples of additives for liquid preparations include acacia, almond oil, ethyl alcohol, fractionated coconut oil, gelatin, glucose syrup, glycerin, hydrogenated edible fats, lecithin, methyl cellulose, methyl or propyl parahydroxybenzoate, propylene glycol, sorbitol, or sorbic acid.
10197] For intravenous (IV) use, a compound according to the invention can be dissolved or suspended in any of the commonly used intravenous fluids and administered by infusion. Intravenous fluids include, without limitation, physiological saline or Ringer's solution. Intravenous administration may be accomplished by using, without limitation, syringe, minipump or intravenous line.
|0198] Formulations for parenteral administration can be in the form of aqueous or non-aqueous isotonic sterile injection solutions or suspensions. These solutions or suspensions can be prepared from sterile powders or granules having one or more of the carriers mentioned for use in the formulations for oral administration. The compounds can be dissolved in polyethylene glycol, propylene glycol, ethanol, com oil, benzyl alcohol, sodium chloride, and/or various buffers.
[0199] For intramuscular preparations, a sterile formulation of a compound or a suitable soluble salt form of the compound, for example the hydrochloride salt, can be dissolved and administered in a pharmaceutical diluent such as Water-for-Inj ection (WFI), physiological saline or 5% glucose. A suitable insoluble form of the compound may be prepared and administered as a suspension in an aqueous base or a pharmaceutically acceptable oil base, e.g., an ester of a long chain fatty acid such as ethyl oleate.
-97-
SUBSTITUTE SHEET ( RULE 26 ) [0200| A dose of an intravenous, intramuscular or parental formulation of a compound may be administered as a bolus or by slow infusion. A bolus is a dose that is administered in less than 30 minutes. In a preferred embodiment, a bolus is administered in less than 15 or less than 10 minutes. In a more preferred embodiment, a bolus is administered in less than 5 minutes. In an even more preferred embodiment, a bolus is administered in one minute or less. An infusion is a dose that is administered at a rate of 30 minutes or greater In a preferred embodiment, the infusion is one hour or greater. In another embodiment, the infusion is substantially constant.
(0201] For topical use the compounds of the present invention can also be prepared in suitable forms to be applied to the skin, or mucus membranes of the nose and throat, and can take the form of creams, ointments, liquid sprays or inhalants, lozenges, or throat paints. Such topical formulations further can include chemical compounds such as dimethylsulfoxide (DMSO) to facilitate surface penetration of the active ingredient.
[0202] For application to the eyes or ears, the compounds of the present invention can be presented in liquid or semi-liquid form formulated in hydrophobic or hydrophilic bases as ointments, creams, lotions, paints or powders.
1 203] For rectal administration the compounds of the present invention can be administered in the form of suppositories admixed with conventional carriers such as cocoa butter, wax or other glyceride.
[0204] Alternatively, the compounds of the present invention can be in powder form for reconstitution in the appropriate pharmaceutically acceptable carrier at the time of delivery. In another embodiment, the unit dosage form of the compound can be a solution of the compound or preferably a salt thereof in a suitable diluent in sterile, hermetically sealed ampoules or sterile syringes. The concentration of the compound in the unit dosage may vary, e.g. from about 1 percent to about 50 percent, depending on the compound used and its solubility and the dose desired by the physician.
[0205] In some embodiments the pharmaceutical compositions described herein are at a therapeutically effective dosage level. In some embodiments, the pharmaceutical composition is at a therapeutically effective levels of between 0.001 to 100 mg/kg. of body weight daily for administration to a patient, e.g., humans and elderly humans. The therapeutically effective amount will generally be about 0.5 mg to 10g per patient per day which may be administered in single or multiple doses. In some embodiments the therapeutically effective amount is between a lower limit of 0.5 mg, 10 mg, 1 mg, 500.0 mg, 1000 mg, 1500 mg, 2000 mg, 2500 mg, 3000 mg, 3500 mg, 4000 mg, 4500 mg, 5000 mg,
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SUBSTITUTE SHEET ( RULE 26 ) 5500 mg, 6000 mg, 6500 mg, 7000 mg, 7500 mg, 8000 mg, 8500 mg, 9000 mg, 9500 mg, and 10000 mg; and an upper limit of 10000 mg, 9500 mg, 9000 mg, 8500 mg, 8000 mg, 7500 mg, 7000 mg, 6500 mg, 6000 mg, 5500 mg, 5000 mg, 4500 mg, 4000 mg, 3500 mg, 3000 mg, 2500 mg, 2000 mg, 1500 mg, 1000 mg, 500.0 mg, 100 mg, 10 mg and 0.5 mg. In some embodiments, the therapeutically effective amount will be about 0.5 mg to 2500 mg per patient per day; in some embodiments about 0.5 mg to 200 mg per patient per day; in some embodiments about 0.5 mg to 500 mg per patient per day; in some embodiments about 0.5 mg to 1000 mg per patient per day; and in yet some other embodiments about 5 mg to 50 mg per patient per day. Pharmaceutical compositions of the present invention may be provided in a solid dosage formulation such as comprising about 0.5 mg to 500 mg active ingredient, or comprising about 1 mg to 250 mg active ingredient. The pharmaceutical composition may be provided in a solid dosage formulation comprising for example about 1 mg, 5 mg, 10 mg, 25 mg, 50 mg, 100 mg, 200 mg, 250 mg, 500 mg or 1000 mg of active ingredient. For oral administration, the compositions may be provided in the form of tablets containing 1.0 to 1000 milligrams of the active ingredient, such as 1, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900, 1000 and 2000 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. The compounds may be administered on a regimen of 1 to 4 times per day, such as once, twice, three times or four times per day.
Unexpected Properties
Mitochondrial uncouplers with improved PK profile
|0206] Some embodiments describe mitochondrial uncouplers depicted herein that have improved pharmacokinetic (PK) profiles, such as markedly increased systemic exposure (i.e. through increased solubility and absorption) consistent with drastically improved potency, and markedly reduced half-lives consistent with a once-daily oral administration regimen and elimination of the toxicity risk factor due to accumulation over prolonged use for chronic conditons or disorders.
[0207] A common feature of prior benzamide compounds with mitochondrial uncoupling activity is low systemic exposure. As shown in Table 1, niclosamide ethanolamine has an oral exposure of merely about 1800 hr*ng/ml in a 24 hr period after 50 mg/kg dosage. Compound 31 in International Patent Publication Number WO 2016/081599 and Compound 27 in US Patent 10,227,315 have systemic exposure of only about 2400
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SUBSTITUTE SHEET ( RULE 26 ) hr*ng/ml after 20 mg/kg oral administration and about 3000 hr*ng/ml after 10 mg/kg oral administration in 24 hr period of time respectively (Table 1). In order to overcome the poor pharmacokinetic properties of low systemic exposure and short half-life of the previously disclosed compounds, studies in animal models required the compound to be mixed with food, and high doses of the compounds were required to achieve efficacy (e g. 1500 ppm niclosamide ethanolamine (NEN) in diet, equivalent to 150 mg/kg/day (Tao, et al. 2014, and WO 2012/068274); or 600-750 ppm (Compound 28 in US10,227,315) in diet, equivalent of 60-75 mg/kg/day ). Neither the high doses required nor the need to mix the compound in food, is compatible with therapeutic development in humans.
[0208| As exemplified in Table 1, the new compounds (e.g., Compounds 2, 9, 78) unexpectedly exhibit significantly increased systemic exposure without compromising uncoupling activity, with 20- to 160- fold increase over the prior uncouplers with the best systemic exposure.
Table 1
SUBSTITUTE SHEET ( RULE 26 )
[0209| Table 2 provides additional compounds with drastically increased systemic exposure (See Example B3).
Table 2
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SUBSTITUTE SHEET ( RULE 26 )
+++: 10,000-30,000; ++++: 30,001- 60,000; +++++: 60,001-120,000; ++++++:>120,000
[0210] As noted in Table 3, Mitochondrial uncoupling activity of Compound 49 in U.S. Patent 10,227,315 is low compared to similar compounds of the present invention.
Table 3:
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SUBSTITUTE SHEET ( RULE 26 )
Drastically Improved Metabolic Stability and Half-life Properties
[0211| Some embodiments describe compounds depicted herein with improved metabolic stability and half-life.
[0212J One key reason previously known mitochondrial uncouplers such as DNP exhibit a narrow therapeutic index is due to their long-half life. For example, DNP has a half- life measured in days to weeks, leading to accumulation of toxic levels in the body during prolonged use to treat chronic conditions or disorders. For development of orally administered drugs, excretion properties are also important. Not only does a drug need to have sufficient exposure, but it also needs to be excreted at a rate that is not too short nor to long. On the one hand, it is important that an oral drug should have sufficiently long half-life in patients so the drug can be administered at certain intervals (e.g. once daily instead of once every hour); on the other hand, the half-life of a compound cannot be too long as this could lead to toxic accumulation of the drug inside the patient. For once-daily orally administered drugs, a half-life around 8-12 hrs is considered a favorable property, while half-lives exceeding 48 hrs are incompatible for once-daily oral administration and could lead to toxic accumulation of the drug. For example, Compound 2 of US Patent 10,227,315 exhibits an extremely long half-life (66.9 hrs, Table 4, Example B8). The long half-life correlated with extreme metabolic stability (rat microsome metabolic stability, half-life 2,131.8 minutes). In
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SUBSTITUTE SHEET ( RULE 26 ) other words, the extremely long half-life of Compound 2 of US Patent 10,227,315 is mainly caused by the liver metabolic enzymes inability to efficiently metabolize these compounds.
|0213] As shown in Table 4, Compound 17 of the present invention unexpectedly has drastically decreased metabolic stability (from 2,132 minutes to 145 minutes, Example B7), and concomitant decreased oral half-life (from 66.9 hrs to 8.6 hrs, Example B8). The structural modification does not decrease the mitochondrial uncoupling activity thus are expected to have better toxicology properties when used to treat chronic conditions or disorders, which require prolonged use .
Table 4
|0214] Half-lives in rat liver microsomal stability assay were determined (Example B7) for a number of compounds described herein and compared with Compound 2, Compound 53, and 54 in U. S. Patent 10,227,315 (Table 5). The compounds of the present invention exhibited drastically improved metabolic stability (favorable half-life less likely to cause toxicity when used to treat chronic conditions or disorders, which require prolonged use) without decreased uncoupling activity (see Table 5 and Table 8).
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SUBSTITUTE SHEET ( RULE 26 ) Table 5
SUBSTITUTE SHEET (RULE 26)
MMP-retaining compounds
10215] Some embodiments describe mitochondrial uncouplers depicted herein that effectively increases OCR without reducing or significantly reducing MMP. These compounds are referred to as MMP -retaining uncouplers in this disclosure.
[0216] Mechanistically, conventional mitochondrial uncouplers transport protons across the mitochondrial inner membrane into the mitochondrial matrix. As mitochondrial membrane potential is supported by proton gradient across the membrane, conventional uncouplers inevitably require rapid dissipation of mitochondrial membrane potential to sustain mitochondrial uncoupling action (Figure 3). Prior to the present disclosure, all mitochondrial uncouplers (herein referred to as conventional uncouplers) tested, had properties that increase OCR and decreased MMP, and the concentrations leading to OCR increase and MMP dissipation correlate (Figure 4). Figure 4 shows a conventional uncoupler, FCCP, has a ratio of CIO%TMRE / Cmin-ocR less than 3, where CIO%TMRE is the concentration leading to 10% MMP retention (or 90% MMP loss, measured by TMRE staining) and
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SUBSTITUTE SHEET ( RULE 26 ) OCR is the minimal concentration leading to OCR increase. As such, the activity for decreasing MMP is considered a second hallmark of mitochondrial uncoupling. As MMP is critical for cell survival and normal functions of cells in many tissues and organs, this seemingly inherent property of conventional mitochondrial uncouplers on dissipating MMP represents a major safety hurdle for therapeutic development.
[0217| In some embodiments, the mitochondrial uncouplers described herein represent a fundamentally different category of mitochondrial uncouplers which effectively induce mitochondrial uncoupling (increase mitochondrial oxygen consumption in the presence of oligomycin) without significantly decreasing mitochondrial membrane potential over a wide concentration range (Figures 5 and 6, Example B2). Figure 5 shows that Compound 25 does not appear to reduce MMP over a wide concentration range where OCR increases and reaches maximal levels. The ratio of CIO%TMRE / Cmin-ocR for Compound 25 is over 25. Figure 6 shows that Compound 64 effectively induces mitochondrial uncoupling without significantly decreasing MMP over a wide concentration range. The ratio of CIO%TMRE / Cmin-ocR for Compound 64 is between 10 to 25.
|0218] After determining the mitochondrial uncoupling activity of a mitochondrial uncoupler described herein (Example Bl), a mitochondrial membrane potential assay was performed on the mitochondrial uncoupler, using the standard TMRE (tetramethylrhodamine ethyl ester) staining method with cultured mammalian cells (Example B2). Results are categorized into three groups, the Compound 25-like compounds are denoted as MMP -Retaining Uncoupling Compounds (CIO%TMRE / Cmin-OCR>25); The Compound 64-like compounds are denoted as MMP -Retaining Uncoupling Compounds (Cio%TMRE / Cmin-ocR between 10 to 25), and Conventional Uncouplers (CIO%TMRE / Cmin-ocR less or equal to 3). The results are summarized in Table 6.
Table 6
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SUBSTITUTE SHEET ( RULE 26 )
CW%TMRE is the concentration leading to 10% MMP retention (or 90% MMP loss, measured by TMRE staining) and Cmin-ocR is the minimal concentration leading to OCR increase
[0219] Mechanistically, chemical uncouplers are lipophilic weak acids or weak bases that localize in mitochondrial inner membrane and transport protons across the membrane into mitochondrial matrix through a protonation and deprotonation cycle (Figure 3 and Figure 7A). As mitochondrial membrane potential (MMP) is supported by proton gradient across the membrane, it is highly unexpected that a compound could uncouple mitochondrial oxidation (transport protons) without significantly decreasing MMP. We analyzed the benzothiazole derivatives of benzamide uncouplers that exhibit MMP -retaining
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SUBSTITUTE SHEET ( RULE 26 ) activity. They all contain a tertiary amine or secondary amine group, which have a pKa over 10.0. The amine groups become positively charged in cellular environment where the pH is generally below 8.0
10220] Without wishing to be bound by theory, we propose the mechanism of action for the MMP- retaining uncouplers, is as illustrated in Figure 7B. In essence, a MMP- retaining uncoupler molecule consists of two functional parts (Figure 7C). The first part functions as a conventional uncoupler, which transports proton from the mitochondrial intermembrane space to the mitochondrial matrix (Figure 7B). The second part is a positively charged functional group that is poorly impermeable to the mitochondrial inner membrane. This feature allows an asymmetrical distribution and orientation of the compounds across mitochondrial inner membrane, with higher concentration of the charged molecules in the intramembrane space than in the mitochondrial matrix, as well as with the positively charged moiety primarily distributed at the outer surface of the membrane (facing the intermembrane space, Figure 7B). As mitochondrial uncoupling occurs and consequently reduction of proton gradient across the membrane, the loss of membrane potential due to proton gradient reduction is compensated by the asymmetrical positive charge distribution across the membrane provided by MMP-retaining uncouplers. Hence, the overall mitochondrial membrane potential is minimally impacted over a wide concentration range of the uncouplers.
[0221 J Figure 7D illustrates the structural feature of the all the MMP-retaining uncouplers listed in Table 6 in which the positive charge is provided by a tertiary amine or a secondary amine. We notice that by modifying the amine containing moiety, one could finetune the MMP-retaining activity, reducing the ratio of Cio%TMRE/Cmin-ocR from >25 to a ratio between 10 to 25. Some modification of the amine-containing moiety could even reduce the ratio close to 3, a ratio equal to that of a typical conventional mitochondrial uncoupler. Figure 7E shows a specific example of the MMP-retaining uncoupler, with the conventional uncoupler component shown in the frame. Improved in vivo safety profiles
|0222] Some embodiments describe MMP- retaining uncouplers exhibit markedly improved safety profiles compared with conventional uncouplers such as the bench mark conventional uncoupler DNP.
[02231 DNP is a conventional mitochondrial uncoupler once used in humans before for obesity treatment. However, it exhibited high levels of toxicity and had a narrow
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SUBSTITUTE SHEET ( RULE 26 ) therapeutic window. It was subsequently withdrawn from market for use in humans. The toxicology profiles of the MMP-retaining uncouplers using Compound 25 and Compound 64 as examples, and compared them with those of DNP. Compounds 25 and Compound 64 have excellent oral absorption, systemic exposure, and efficacy in reducing blood glucose and liver steatosis in animal models (Tables 2, 5, and 9).
I0224| Some embodiments show MMP-retaining uncouplers exhibit markedly improved acute toxicity profiles. Figure 8 (Example B9) shows Compound 64 and Compound 25 have LD50 (lethal dose, 50% animal death)/MED (minimal efficacious dosage) ratios over 400 and 200 respectively, while the LD50/MED ratio of DNP is 30.
[02251 Some embodiments show MMP-retaining uncouplers exhibiting markedly improved short-term (10-day) toxicology profiles. Figure 9-10 (Example B10) show that the ratio ofNOAELs (no-observable-adverse-effect-level) over MED of Compounds 64 is over 40 (between 40-57 using different parameters), NOAEL/MED ratio of Compound 25 is over 19, while NOAEL/MED ratio of DNP is reportedly less than 3.
[0226 [ Some embodiments describe a method of preparing MMP-retaining uncouplers from conventional mitochondrial uncouplers by adding a positively charged side chain to the conventional uncouplers, as illustrated in Figure 7C, for example, by adding a tertiary amine (or secondary amine) containing side chain (Figure 7D). As shown in Figure 7E, a tertiary amine-containing moiety was added to a conventional uncoupler (the structure in the Frame). This modification effectively converts the conventional uncoupler (with the ratio of Cio%TMRE/Cmin-ocR of 3) to an MMP-retaining uncoupler (listed in Table 2, with the ratio of Cio%TMRE/Cmin-ocROver 25).
[0227] Some embodiments describe a mitochondrial membrane-retaining uncoupler compound of the Formula:
(RA)U-RB; or a pharmaceutically acceptable salt, solvate or prodrug thereof; wherein RAand RB are covalently linked; each RA is independently a moiety containing a secondary or tertiary amine; u is an integer selected from the group consisting of 1 and 2; and
RB is a conventional mitochondrial uncoupler prior to being covalently linked to RA; provided the mitochondrial membrane-retaining uncoupler compound is not
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SUBSTITUTE SHEET ( RULE 26 )
[0228] Some embodiments describe a method of preparing a mitochondrial membrane-retaining mitochondrial uncoupler comprising
1. identifying a conventional mitochondrial uncoupler;
2. designing a compound that covalently links at least one moiety capable of becoming positively charge in a cellular environment, to a conventional mitochondrial uncoupler; and
3. preparing the compound of step 2, wherein the compound is a mitochondrial membrane retaining uncoupler compound.
In some embodiments the moiety capable of becoming positively charge in a cellular environment is a secondary or tertiary amine moiety.
[0229| Some embodiments describe a method of preparing a mitochondrial membrane-retaining mitochondrial uncoupler comprising
1. identifying a conventional mitochondrial uncoupler;
2. designing a compound that covalently links at least one secondary or tertiary amino moiety to a conventional mitochondrial uncoupler; and
3. preparing the compound of step 2, wherein the compound is a mitochondrial membrane retaining uncoupler compound.
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SUBSTITUTE SHEET ( RULE 26 ) [0230 ] In some embodiments the secondary or tertiary amino moiety or RA is selected from the group consisting of -(CH2)NH(CH2)2OCH3, -CH2NHC(O)CH3, - previously described in any embodiment described herein.
[02311 In some embodiments the secondary or tertiary amino moiety or RA is selected from the group consisting of -CH2NHSO2CH3, -CH2N(CH3)2, -(CH2)2N(CH3)2, -
-112-
SUBSTITUTE SHEET ( RULE 26 )
[02321 In some embodiments the secondary or tertiary amino moiety or RA is
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SUBSTITUTE SHEET ( RULE 26 )
[0233} In some embodiments the conventional mitochondrial uncoupler or RB is selected from the group consisting of a conventional mitochondrial uncoupler as described herein. In some embodiments the conventional mitochondrial uncoupler or RB is selected from the group consisting of a conventional mitochondrial uncoupler as described in: U. S. Patent No. 10,227,3158, U. S. Patent Application No. 15/527,808 and Childress, E. S., et al. (2018) i Med. Chem., 61(11), 4641- 4655.
[0234} In some embodiments the conventional mitochondrial uncoupler or RB is selected from the group consisting of
[0235| In some embodiments the secondary or tertiary amino moiety or RA; the conventional mitochondrial uncoupler or RB; and the mitochondrial membrane-retaining uncoupler compound (RA)U-RB are as described in Table 7.
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SUBSTITUTE SHEET ( RULE 26 ) Table 7
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SUBSTITUTE SHEET (RULE 26)
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SUBSTITUTE SHEET (RULE 26)
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SUBSTITUTE SHEET (RULE 26)
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SUBSTITUTE SHEET (RULE 26)
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SUBSTITUTE SHEET (RULE 26)
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SUBSTITUTE SHEET (RULE 26)
SUBSTITUTE SHEET (RULE 26) Methods of Treatment
10236] In some embodiments, a method of treating a mitochondria-related condition or diorder, in a subject in need thereof, comprises administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition according to embodiments described herein.
[0237] In some embodiments, the mitochondria-related condition or disorder has and one or more underlying causal factors selected from the group consisting of hyperglycemia, abnormal accumulation of lipid in cells, abnormal accumulation of lipid in tissues, abnormal lipid metabolism, insulin resistance, abnormal cell proliferation, abnormal TGF-beta activation, and abnormal fibrosis. In some embodiments, the mitochondria-related condition or disorder has and one or more underlying causal factors selected from the group consisting of hyperglycemia, lipid accumulation, insulin resistance, altered cellular metabolism, fibrosis, aberrant TGF-beta activation, or aberrant cell proliferation.
[0238] In some embodiments, the mitochondria-related condition or disorder has and one or more underlying symptoms selected from the group consisting of hyperglycemia, abnormal accumulation of lipid in cells, abnormal accumulation of lipid in tissues, abnormal lipid metabolism, insulin resistance, abnormal cell proliferation, abnormal TGF-beta activation, and abnormal fibrosis. In some embodiments, the mitochondria-related condition or disorder has and one or more underlying causal factors selected from the group consisting of hyperglycemia, lipid accumulation, insulin resistance, altered cellular metabolism, fibrosis, aberrant TGF-beta activation, or aberrant cell proliferation.
[0239] In some embodiments, the mitochondria-related condition or disorder is a metabolic disease, cancer, an autoimmune disease, pulmonary fibrosis, a dermatological disorder, an infectious disease or a neurodegenerative disease. In some embodiments, the mitochondria-related condition or disorder is a metabolic disease, cancer, and autoimmune disease, or infectious disease.
[02401 In some embodiments, the mitochondria-related condition or disorder is a metabolic disease. In some embodiments the metabolic disease is selected from the group consisting of type 2 diabetes, a disease characterized by insulin resistance or hyperglycemia; obesity or obesity related complications, and a disease characterized by abnormal lipid accumulation.
[0241] In some embodiments, a method of treating a metabolic disease or disorder characterized by insulin resistance or by abnormal accumulation of lipid in tissue, or a
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SUBSTITUTE SHEET ( RULE 26 ) disease or a disorder in which insulin resistance or abnormal accumulation of lipid in tissue is a symptom, or treating cancer or hyperplasia, in a subject in need thereof, comprises administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition according to embodiments described herein. In some embodiments, the metabolic disease or disorder is type 2 diabetes, or a disease characterized by insulin resistance or hyperglycemia.
[0242] In some embodiments, a metabolic disease or disorder described in any embodiment herein is a complication caused by type 2 diabetes, selected from the group consisting of diabetes-induced cardiovascular diseases, neurodegenerative disorders, atherosclerosis, hypertension, coronary heart diseases, nephropathy, retinopathy, neuropathy, and diabetic heart failure. In some embodiments, the metabolic disease or disorder is obesity or obesity related complications.
[0243] In some embodiments, a metabolic disease or disorder described in any embodiment herein is non-alcoholic fatty liver disease (NAFLD), comprising at least one prognosis stage of this disease selected from the group consisting of hepatic steatosis, nonalcoholic steatohepatitis (NASH), cirrhosis, and NAFLD induced hepatocellular carcinoma (HCC). In some embodiments, the metabolic disease or disorder is alcoholic fatty liver disease, or a complication caused by alcoholic fatty liver diseases. In some embodiments, the complication of alcoholic fatty liver disease comprises alcoholic hepatitis, cirrhosis, or a combination thereof.
[0244] In some embodiments, the metabolic disease or disorder is dyslipidemia, or a complication caused by dyslipidemia.
[0245] In some embodiments, the cancer is a primary cancer selected from the group consisting of hepatocellular carcinoma, colorectal carcinoma, pancreatic cancer, breast cancer, prostate cancer, leukemia, lymphoma, melanoma, ovarian cancer, and lung cancer. In some embodiments, the cancer is a metastatic cancer originated from a primary tumor of other tissue types. In some embodiments, the metastatic sites are selected from the group consisting of liver, lung and the intraperitoneal cavity.
[0246] In some embodiments, the compound of embodiments described herein is administered in combination with a second agent indicated for the above-mentioned disorders or diseases, either concomitant with, prior to, or after the administration of the second agent. In some embodiments, the second agent is an anti-diabetic agent selected from the group consisting of metformin, insulin, insulin analogs, sulfonylureas, biguanides, meglitinides, thiazolidinediones, alpha glucosidase inhibitors, GLP-1 agonists, SGLT2 inhibitors and DPP-
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SUBSTITUTE SHEET ( RULE 26 ) 4 inhibitors. In some embodiments, the second agent is an anti-obesity agent. In some embodiments, the second agent is an anti -nonalcoholic fatty liver disease agent. In some embodiments, the second agent is anti-alcoholic fatty liver disease agent. In some embodiments, the second agent is an anti-dyslipidemia agent.
[0247 j In some embodiments, a compound of embodiments described herein, is administered in combination with a second anti- non-alcoholic fatty liver disease agent. In some embodiments, a compound of the invention is administered in combination with a second anti- alcoholic fatty liver disease agent. In some embodiments, a compound of the invention is administered in combination with a second anti- dyslipidemia agent.
[02481 In some embodiments, the compound may be administered in combination with a second anti-cancer agent or anti-cancer regimen. In some embodiment the second anti-cancer agent s an immunoncological agent. In some embodiments the immunocological agent is selected from the group consisting of an antibody against PD-1/PD-L1, an antibody against other immune check point proteins, CAR-T cells, and other therapeutic immune cells. In some embodiments, the compound may be administered prior to, concomitantly with, or subsequently to administration of the second anti-metabolic disease or anti-cancer agent.
|0249] In some embodiments, the subject is a mammalian animal. In some embodiments, the subject is a human. In some embodiments, the compound described herein is used as a veterinarian drug to treat diabetes or a diabetes-associated disease, and the subject is a mammalian animal.
[0250] Some embodiments are directed to a method for long-term disease management of a metabolic disease or disorder comprising administering to a subject in need of such long-term management an effective amount of a compound or a pharmaceutical composition described herein. In some embodiments, a method for long-term disease management of a metabolic disease or disorder, or for long-term disease management of cancer, comprises administering to a subject in need of such long-term management an effective amount of a compound or a pharmaceutical composition according to any of the embodiments described herein. In some embodiments, the metabolic disease or disorder is obesity, obesity -related complications, type 2 diabetes, or type 2 diabetes related complications. In some embodiments, the cancer is any primary tumor or metastatic tumor.
|0251] In some embodiments, the present disclosure describes the use of a compound according to any of the embodiments described herein in the manufacture of a medicament for treatment of diabetes, obesity, non-alcoholic fatty liver disease, alcoholic fatty liver disease, dyslipidemia, or a disease where insulin resistance or abnormal lipid
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SUBSTITUTE SHEET ( RULE 26 ) accumulation in tissue is a symptom, or related disorders or complications, including, but not limited to, hepatic steatosis, non-alcoholic steatohepatitis (NASH), cirrhosis, or NAFLD induced hepatocellular carcinoma (HCC). In some embodiments, the compound of embodiments herein may be used to manufacture a medicament for the treatment of cancer, a disease where cell proliferation (hyperplasia) is a symptom, or cancer or hyperplasia related complications.
[0252J Some embodiments herein are directed to a method of treating or preventing a metabolic disease or disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein.
[0253] Some embodiments herein provide for a method of treating and alleviating the symptoms of obesity (characterized by excessive accumulation of lipid in adipocytes), pre-type 2 diabetes (characterized by insulin resistance usually caused by ectopic accumulation of lipid in cells of liver and muscle), type 2 diabetes (characterized by insulin resistance and hyperglycemia), non-alcoholic fatty liver diseases or alcoholic fatty liver disease (characterized by abnormal accumulation of lipid in liver), dyslipidemia (characterized by abnormal lipid deposit in tissue other than adipose), and one or more complications of the above mentioned metabolic disorders, including, but not limited to, hypertension, cardiovascular diseases, nephropathy, and neuropathy comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein. These diseases or disorders may be caused by dietary, environmental, medical and/or genetic factors. The methods described herein may also be used for prevention of the above-mentioned metabolic diseases for a subject with risk factors including, but not limited to, dietary, environmental, medical, and genetic predispositions. In addition, some embodiments provide a method for long-term chronic disease management and longevity management by reducing insulin resistance or reducing glucose levels in the blood.
[0254] In some embodiments, the metabolic disease or disorder is type 2 diabetes, or related diseases leading to insulin resistance or hyperglycemia. In some embodiments, the metabolic disease or disorder is obesity or one or more obesity related complications.
10255] In some embodiments, the metabolic disease or disorder is non-alcoholic fatty liver disease, (NAFLD), including nonalcoholic steatohepatitis (NASH) and cirrhosis, or alcoholic fatty liver disease (AFLD). In some embodiments, the metabolic disease or disorder
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SUBSTITUTE SHEET ( RULE 26 ) is hepatic steatosis, non-alcoholic steatohepatitis (NASH), cirrhosis, or NAFLD induced hepatocellular carcinoma (HCC).
|0256] In some embodiments, the metabolic diseases or disorder is one or more complications of type 2 diabetes including, but not limited to, type 2 diabetes induced hypertension, cardiovascular disease, nephropathy, atherosclerosis, dyslipidemia, retinopathy, neurodegenerative disorders, diabetic heart failure, and neuropathy. In some embodiments, the metabolic disease or disorder is pre-type 2 diabetes. In some embodiments, the metabolic disease or disorder is dyslipidemia.
[0257] In some embodiments, the disease to be treated may be a mitochondrial disorder. In some embodiments, the metabolic disorder may be LHON (leber heredity optic neuropathy), MELAS (mitochondrial myopathy, mitochondrial encephalomyopathy, lactic acidosis and stroke-like episodes), MERRF (myoclonic epilepsy and ragged red muscle fiber), Leigh Syndrome, MILS (maternally inherited Leigh Syndrome), NARP (neurogenic muscle weakness, ataxia and retinitis pigmentosa), FBSN (familial bilateral striatal necrosis), or KSS (Kearns Sayre Syndrome).
[0258] Some embodiments are directed to a method of treating or preventing cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein. In some embodiments, the cancer may be primary cancer or metastatic cancer. In some embodiments, the cancer is primary cancer including but not limited to hepatocellular carcinoma, colorectal carcinoma, pancreatic cancer, breast cancer, prostate cancer, leukemia, lymphoma, melanoma, ovarian cancer, lung cancer. In some embodiments, the cancer is metastatic liver cancer originated from the primary tumor of other tissue types. In some embodiments, the cancer is metastatic lung cancer originated from the primary tumor of other tissue types. In some embodiments, the cancer is metastatic cancer to other sites including intraperitoneal cavity.
[0259] Some embodiments are directed to a method of treating or preventing autoimmune diseases in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein. In some embodiments, the autoimmune disease is celiac disease, diabetes mellitus type 1, Graves' disease, inflammatory bowel disease, multiple sclerosis, psoriasis, rheumatoid arthritis, and systemic lupus erythematosus.
[0260] Some embodiments are directed to a method of treating or preventing a dermatological disorder in a subject in need thereof, comprising administering to the subject a
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SUBSTITUTE SHEET ( RULE 26 ) therapeutically effective amount of a compound or pharmaceutical composition described herein. In some embodiments, the dermatological disorder is eczema, dyshidrotic eczema, seborrheic eczema psoriasis, rosacea, dermatitis and atopic dermatitis.
|0261] Some embodiments are directed to a method of treating or preventing an infectious disease of non-viral parasites in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein. In some embodiments, the infectious disease is a viral infections. In some embodiments the viral infection is an envelope viral infection. In some embodiments the viral infection is selected from the group consisting of SARS- CoV-2 , a corana virus infection and an Ebola viral infection.
[0262] In some embodiments, the disease to be treated may be a heart disorder comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein. In some embodiments, the heart disorder may be hypertension or cardiovascular disease. In some embodiments, the disease to be treated may be a central nervous system (CNS) disease. In some embodiments, the CNS disease may be stroke, Alzheimer’s, Parkinson’s, Huntington’s, or ALS (amyotropic lateral sclerosis).
[0263] In some embodiments, the disease to be treated may be a disorder associated with increased ROS (reactive oxygen species) production comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein. Increased ROS has been associated with aging, Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, ALS (amyotropic lateral sclerosis), mitochondrial diseases, and various cancers.
[02641 The compounds and pharmaceutical compositions described herein may be administered by oral, parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV, intracistemal injection or infusion, subcutaneous injection, or implant), by inhalation spray, ophthalmic, nasal, vaginal, rectal, sublingual, or topical routes of administration and may be formulated, alone or together, in suitable dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles appropriate for each route of administration. The compounds and pharmaceutical compositions described herein may also be formulated as a controlled-release formulation.
[0265] The compounds described herein may be administered topically and can be formulated into a variety of topically administrable pharmaceutical compositions comprising an active ingredient and a dermatologically acceptable base and/or an ophthalmically
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SUBSTITUTE SHEET ( RULE 26 ) acceptable base. Such pharmaceutical compositions can be formulated, e.g., as solutions, suspensions, spray, lotions, gels, pastes, medicated sticks, balms, shampoos, soap bars, liquid soaps, creams or ointments. In one embodiment, the pharmaceutical composition is the form of an ointment that can be applied in or around the eye of a mammal, including a human.
[0266] In some embodiments, a dermatologically and/or ophthalmically acceptable base includes a pharmaceutically acceptable ointment base. Examples of suitable ointment bases include, but are not limited to oleaginous ointment bases such as petrolatum (e.g., liquid petrolatum or white petrolatum), plastibase, hard paraffin, white soft paraffin, yellow soft paraffin, liquid paraffin, emulsifying wax, microcrystalline wax, white bees wax, yellow bees wax, carnauba wax, wool wax (wool fat), mineral oil, olive oil, purified lanolin, anhydrous lanolin, and water soluble ointment bases such as polyethylene glycol (e.g., polyethylene glycol 400 or polyethylene glycol 3350), propylene glycol, polyoxyethylene, polyoxypropylene, or any combinations thereof.
[0267] In some embodiments, a dermatologically and/or ophthalmically acceptable base includes one or more polymers as suspending agents. Useful polymers include, but are not limited to, water-soluble polymers such as cellulosic polymers, e g., hydroxypropyl methylcellulose, and water-insoluble polymers such as cross-linked carboxyl- containing polymers. A dermatologically and/or ophthalmically acceptable base can also include a dermatologically and/or ophthalmically acceptable mucoadhesive polymer, e.g., carboxymethylcellulose, carbomer (acrylic acid polymer), carbopol (copolymers or acrylic acid crosslinked with a polyakenyl polyether), poly(methylmethacrylate), polyacrylamide, polycarbophil, acrylic acid/butyl acrylate copolymer, sodium alginate, or dextran.
[0268] In some embodiments, a dermatologically and/or ophthalmically acceptable base includes one or more viscosity enhancing agents. Examples of suitable viscosity enhancing agents include, but are not limited to, methyl cellulose, xanthan gum, gum tragacanth, carboxymethyl cellulose, silica, silicone, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, hydroxypropylmethyl cellulose acetate stearate, hydroxypropylmethyl cellulose phthalate, carbomer, polyvinyl alcohol, alginates, acacia, chitosans, acacia, com starch, gelatin, or combinations thereof.
[0269] In some embodiments, a dermatologically and/or ophthalmically acceptable base includes one or more dermatologically and/or ophthalmically acceptable pH adjusting agents or buffering agents, including, but not limited to, acids such as acetic, boric, citric, lactic, phosphoric and hydrochloric acids; bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium, lactate and tris-
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SUBSTITUTE SHEET ( RULE 26 ) hydroxymethylaminomethane; and buffers such as citrate/dextrose, sodium bicarbonate and ammonium chloride. Such acids, bases and buffers are included in an amount required to maintain pH of the composition in a dermatologically and/or ophthalmically acceptable range.
[0270 j In some embodiments, a dermatologically and/or ophthalmically acceptable base includes one or more dermatologically and/or ophthalmically acceptable salts in an amount required to bring osmolality of the composition into a dermatologically and/or ophthalmically acceptable range. Such salts include, but are not limited to, those having sodium, potassium or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate or bisulfite anions; specific salts include, e.g., sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite, and ammonium sulfate.
[0271] In some embodiments, a dermatologically and/or ophthalmically acceptable base includes one or more dermatologically and/or ophthalmically acceptable preservatives to inhibit microbial activity. Suitable preservatives include, but are not limited to, mercury-containing substances such as merfen and thiomersal; stabilized chlorine dioxide; and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide and cetylpyridinium chloride.
[02721 In further embodiments, a dermatologically and/or ophthalmically acceptable base includes one or more dermatologically and/or ophthalmically acceptable surfactants to enhance physical stability, or for other purposes. Suitable nonionic surfactants include isohexadecane, cyclomethicone, copolymers of ethylene glycol and propylene glycol, polyoxyethylene fatty acid glycerides and vegetable oils, e.g., polyoxyethylene (60) hydrogenated castor oil; and polyoxyethylene alkylethers and alkylphenyl ethers, e.g., octoxynol 10, octoxynol 40.
|0273| In further embodiments, a dermatologically and/or ophthalmically acceptable base includes one or more dermatologically and/or ophthalmically acceptable penetration enhancers to enhance physical stability, or for other purposes. Penetration enhancers are substances which enhance passage of topically-applied compounds into the stratum, comeum of the skin and therefrom into the epidermis and dermis. Examples include, but are not limited to: dimethyl isosorbide, ethoxy diglycol, 1- dodecylazacycloheptan-2-one, propylene glycol, oleyl alcohol, polyoxyethylene ester, sorbitan mono-9-octadecenoate, poly(oxy-l,2-ethanediyl) and derivatives thereof, ethanol, glyceryl monoethyl ether, monoglycerides, isopropylmyristate, lauryl alcohol, lauric acid, lauryl lactate, terpinol, menthol, D-limonene, beta-cyclodextrin, DMSO (dimethyl sulfoxide),
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SUBSTITUTE SHEET ( RULE 26 ) polysorbates, fatty acids (e g., oleic), bile salts, N-methylpyrrolidone, polyglycosylated glycerides, l-dodecylazacycloheptan-2-one (Azone®), Cyclopentadecalactone (CPE-215®), Alkyl -2-(N,N-disubstituted amino)-alkanoate ester (NexAct®), 2-(n-nonyl)- 1,3 -di oxolane (DEP A®), and penetration enhancers shown for example in U.S. Pat. Nos. 3,909,816; 4,405,616; 4,801,586; 4,861,764; 4,886,783; 4,983,396; 5,118,845; 5,196,410, 8486,374 and 8,741,265, each of which is hereby expressly incorporated herein by reference in its entirety.
[0274] In further embodiments, a dermatologically and/or ophthalmically acceptable base includes one or more dermatologically and/or ophthalmically acceptable permability enhancers to enhance physical stability, or for other purposes. A variety of classes of compounds may serve as suitable permeability enhancers according to the invention. A first category includes fatty acids and salts and esters thereof, including mono-, di-, and triglycerides. Medium chain length fatty acids, especially C8 and CIO acids, and their salts and esters are particularly useful. Suitable specific examples include sodium caprylate, sodium caprate, CAPMUL® glycerides (available from Abitec of Columbus, Ohio), LABRASOL® glycerides (PEG-8 caprylic/capric glycerides, available from Gattefosse SAS of Saint Priest, Cedex, France), GELUCIRE® 44/14 (PEG-32 glyceryl laurate EP, available from Gattefosse), other glycerides & fatty acid esters, CREMOPHOR® (BASF, Ludwigshafen, Germany), D-a-tocopheryl polyethylene glycol 1000 succinate, vegetable oils, polyoxylglycerides, and medium chain mono- and diacylglycerides.
[0275] One example of this class, CAPMUL® MCM L8 (glycerol monocaprylate) (available from Abitec of Columbus, Ohio), is composed of mono- and diglycerides of medium chain fatty acids (mainly caprylic, with some capric) and 7% maximum free glycerol. It contains at least 44% alpha monoglycerides (as caprylate).
[0276| Other examples of this class of enhancers include GATTEFOSSE compositions 61 A through 61H which are proprietary to Gattefosse SAS, but generally are composed of mixtures containing one or more of medium chain mono-, di-, or triglycerides, polysorbate derivatives, polyoxyl castor oil derivatives, polyethylene glycol derivatives including polyethylene glycol glycerides, polyoxyl ethers, vegetable oils, glycerin, and similar GRAS (generally regarded as safe) lipidic components in varying amounts. These components are part of individual commercial products such as CAPRYOL™ 90, CAPRYOL™ PGMC, LAUROGLYCOL™ 90, GELUCIRE® 44/14, Plurol Oleique CC497, LABRASOL®, LABRAFIL® M1944CS (apricot kernel oil PEG-6 esters), Transcutol HP, Peceol, and Maisine 35-1, all of which are available from Gattefosse SAS.
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SUBSTITUTE SHEET ( RULE 26 ) [0277| While not falling directly within this class, glycerol itself has been found to impart excellent permeability enhancement, particularly for neuraminidase inhibitors. This result was not anticipated as glycerol is not considered a permeability enhancer.
|0278] A second category of enhancers includes surfactants having a steroidal structure, such as bile acid salts. Examples of suitable compounds include sodium cholate, sodium deoxycholate, glycocholate, glycoursodeoxycholate, taurocholate, taurodeoxycholate, and steroid detergents/bile salts. Other surfactants may also be suitable permeability enhancers, including cationic, anionic, and nonionic surfactants. Examples include polysorbate 80, hexadecyldimethylbenzylammonium chloride, N-hexadecylpyridinium bromide, dodecyltrimethylammonium bromide, hexadecyltrimethylammonium bromide, tetradecyl-P-D-maltoside, octylglucoside, glycyrrhetinic acid, 3-(N,N- dimethylpalmitylammonio)propane-sulfonate, and sodium lauryl sulfate.
[0279] Cyclodextrins may also be used as suitable enhancers. Examples include P-cyclodextrin, hydroxypropyl-P-cyclodextrin, y-cyclodextrin, and hydroxypropyl-y- cyclodextrin.
10280] A variety of other compounds may also be used as enhancers. Examples include sodium salicylate, ethylenediamine tetraacetic acid (EDTA), citric acid, chitosan & chitosan derivatives, N-trimethyl chitosan chloride, monocarboxymethyl-chitosan, palmitoyl carnitine chloride, acyl carnitines, ethylene glycol tetraacetic acid (EGTA), 3-alkylamido-2- alkoxypropyl-phosphocholine derivatives, alkanoylcholines, N-acetylated amino acids (based on a- and non-a-amino acids), mucoadhesive polymers, phospholipids, piperine, 1- methylpiperazine, a-amino acids, and mineral oil.
[0281] Thus a wide variety of enhancer compounds may be selected from the group consisting of fatty acids, fatty acid esters, fatty acid salts, glycerol, surfactants, cyclodextrins, sodium salicylate, ethylenediamine tetraacetic acid, citric acid, chitosan, chitosan derivatives, N-trimethyl chitosan chloride, monocarboxymethyl-chitosan, palmitoyl carnitine chloride, acyl carnitines, ethylene glycol tetraacetic acid, 3-alkylamido-2- alkoxypropyl-phosphocholine derivatives, alkanoylcholines, N-acetylated amino acids, mucoadhesive polymers, phospholipids, piperine, 1 -methylpiperazine, a-amino acids, and mineral oil.
10282] The permeability enhancer and the polar agent may be mixed in any proportion so long as there is provided a therapeutically effective amount of the polar agent and a permeability-enhancing amount of the enhancer compound. Enhancement in dermal bioavailability of topically administered polar agents can depend on the nature and
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SUBSTITUTE SHEET ( RULE 26 ) concentration of the enhancer compound with which the agent is formulated. It is thus contemplated that the required therapeutic amount may be contained in a single dosage form or divided between one or more dosages intended for application at the same time or in sequence.
[0283 j The permeability enhancers act relatively independently of the concentration of polar agent. Differing permeability enhancers can reach either optimal or maximum enhancement over a wide concentration range depending on their particular inherent enhancement potential. Often, enhancers have a non-linear dose response relationship between concentration of enhancer present and amount of increased polar agent absorption. The amount of enhancer to be utilized in an oral dosage form with a polar agent is initially based upon the enhancement properties observed in Caco-2 cell assays at varying fixed enhancer concentrations. Based upon those results, an effective in vivo amount of enhancer compound for a human formulation can be estimated, demonstrated and optimized without undue experimentation using methods well known to those skilled in the formulation art, to achieve a desired pharmacokinetic in vivo profile.
[0284] In formulating the composition of this invention, it will be apparent to those skilled in the formulation art that more effective enhancer compounds would require less polar agent than less effective permeability enhancers to achieve a target pharmacokinetic profile. Given those considerations and variations, the amount of enhancer may be at least about 0.1 wt % of the combined weight of enhancer and polar agent, more preferably at least about 50 wt %, and more preferably at least 70 wt % of the combined weight of enhancer and polar agent. The amount is preferably at most 95 wt %, more preferably at most 80 wt %, and more preferably at most 75 wt % of the combined weight of the enhancer and polar agent. Thus, as shown in the examples, a typical dosage form may contain a wide range of concentrations of enhancer compounds depending on the compound itself and its efficacy in enhancing the permeability of polar agents following oral administration. Concentrations as low as 0.001% by weight up to 20% have been demonstrated to be effective in enhancement of the permeability of polar agents.
[0285] In yet other embodiments, a dermatologically and/or ophthalmically acceptable base includes one or more antioxidants to enhance chemical stability where required. Suitable antioxidants include, by way of example only, butylated hydroxytoluene (BHT), sodium ascorbate, ascorbic acid, sodium metabisulfite, and tocopherol. In certain embodiments, antioxidants enhance chemical stability where required.
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SUBSTITUTE SHEET ( RULE 26 ) [0286] In addition to those enumerated above, any other surfactant, moisturizer, gelling agent, preservative, colorant or pigment, antioxidant, radical scavenger, emulsifier, humectant, pH modifier, chelating agent, or other dermatologically acceptable excipient commonly known to those of ordinary skill in the art as useful in topical compositions is contemplated as useful in the compositions described herein. Further, any non-toxic, inert, and effective topical carrier may be used to formulate the compositions described herein.
[0287] Well-known carriers used to formulate other topical therapeutic compositions for administration to humans will be useful in these compositions. Examples of such components that are well known to those of skill in the art are described in The Merck Index, Thirteenth Edition, Budavari et al., Eds., Merck & Co., Inc., Rahway, N.J. (2001); the CTFA (Cosmetic, Toiletry, and Fragrance Association) International Cosmetic Ingredient Dictionary and Handbook, Tenth Edition (2004); and the "Inactive Ingredient Guide", U.S. Food and Drug Administration (FDA) Center for Drug Evaluation and Research (CDER) Office of Management, http://www.accessdata.fda.gov/scripts/cder/iig/index.cfm, the contents of which are hereby incorporated by reference in their entirety. Examples of such useful pharmaceutically acceptable excipients, carriers and diluents include distilled water, physiological saline, Ringer's solution, dextrose solution, Hank's solution, and DMSO, which are among those preferred for use herein.
[0288] These additional other inactive components, as well as effective formulations and administration procedures, are well known in the art and are described in standard textbooks, such as Goodman and Gillman's: The Pharmacological Bases of Therapeutics, 8th Ed., Gilman et al. Eds. Pergamon Press (1990) and Remington's Pharmaceutical Sciences, 17th Ed., Mack Publishing Co., Easton, Pa. (1990), both of which are incorporated by reference herein in their entirety.
[0289] The composition may be used immediately or stored for later use in any type of container known to one of skill in the art such as, for example, pouch, jar, bottle, tube, ampule and pre-filled syringe. Finally, the composition may be sterilized by any method known to one of skill in the art such as, for example, y radiation.
[0290] The compounds and pharmaceutical compositions described herein may be administered at prophylactically effective dosage levels to prevent the above-recited conditions and disorders, as well as to prevent other conditions and disorders characterized by insulin resistance or hyperglycemia.
[0291] The pharmaceutical compositions and compounds of embodiments herein can be administered in a wide range of dosage-forms including, for example, solid dosage
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SUBSTITUTE SHEET ( RULE 26 ) forms and liquid dosage forms. Solid dosage forms may include powders, tablets, pills, capsules, suppositories, or dispersible granules. A solid carrier can be one or more substances that function as a diluting agent, flavor additive, solvent, lubricant, suspension agent, binder, preservative, tablet-disintegrating substance or encapsulating material. In powdered form, the carrier may be a finely pulverized solid including lactose, hydroxypropylmethylcellulose and PVP, mixed with an appropriate amount of the active ingredient. Appropriate carriers for powder and tablet forms include for example magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, stiffeners, gelatins, tragacanth, methylcellulose, and sodium carboxymethylcellulose.
[02921 Liquid dosage forms include for example solutions, suspensions, and emulsions. Also included are pharmaceutical compositions in solid form that are meant to be converted to liquid form shortly prior to consumption. These forms may include, in addition to the active ingredients, artificial colors, flavors, stabilizers, buffers, natural or artificial sweeteners, dispersing agents, thickeners, dissolving agents and the like.
[0293| Solutions or mixtures may be administered directly to the nasal cavity using conventional means, such as drops or sprays. The pharmaceutical composition may be produced in individual or multi-dose forms. Multi-dose forms would include a dropper, pipette or atomizer that delivers a predetermined volume of the pharmaceutical composition. [0002] The pharmaceutical compositions and compounds of embodiments herein may be provided in individual dosage units that contain a suitable amount of the active ingredient. The individual doses may be provided in a package, or as a kit that includes a measuring device, e.g., a device for measuring oral or injectable dosages (i.e., a measuring cup, needle, or syringe). The kit can also include, other materials such buffers, diluents, filters, and package inserts with instructions for use. A label may be present on the on the kit to indicate that the pharmaceutical composition is used for a specific therapy, and may also indicate directions for use.
[0294] If desired, the pharmaceutical compositions of the present invention may further comprise one or more additional active agents. Where it is appropriate, any of the active agents may be administered in the form of the compound per se, and/or in the form of a salt, polymorph, ester, amide, prodrug, derivative, or the like, provided the salt, polymorph, ester, amide, prodrug or derivative is suitable pharmacologically. Where it is appropriate, salts, esters, amides, prodrugs and other derivatives of the active agents may be prepared using standard procedures known to those skilled in the art of synthetic organic chemistry and described, for example, by J. March, Advanced Organic Chemistry: Reactions, Mechanisms
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SUBSTITUTE SHEET ( RULE 26 ) and Structure, 4th Ed. (New York: Wiley-Interscience, 1992). For any active agents that may exist in enantiomeric forms, the active agent may be incorporated into the present pharmaceutical compositions either as the racemate or in enantiomerically enriched form.
|0295] The dosage of the active compound(s) being administered will depend on the condition being treated, the particular compound, and other clinical factors such as age, sex, weight, and health of the subject being treated, the route of administration of the compound(s), and the type of pharmaceutical composition being administered (tablet, gel cap, capsule, solution, suspension, inhaler, aerosol, elixir, lozenge, injection, patch, ointment, cream, etc.) It is to be understood that the present disclosure has application for both human and animal use. The amount of the compound, or an active salt or derivative thereof, required for use in treatment will be ultimately at the discretion of the attendant physician or clinician.
|0296] As described above, the compounds of the invention are useful for the prevention, treatment, control, amelioration, or reduction of risk of the diseases, disorders and conditions noted herein. The dosage of the compound as an active ingredient in the pharmaceutical compositions of this invention may be varied so that a suitable dosage form is obtained. The active ingredient may be administered to patients (animals and human) in need of such treatment in dosages that will provide optimal pharmaceutical efficacy. The selected dosage depends upon the desired therapeutic effect, on the route of administration, and on the duration of the treatment. The dose will vary from patient to patient depending upon the nature and severity of disease, the patient's weight, special diets then being followed by a patient, concurrent medication, and other factors which those skilled in the art will recognize. Generally, dosage levels of between 0.001 to 100 mg/kg. of body weight daily are administered to the patient, e.g., humans and elderly humans. The therapeutically effective amount will generally be about 0.5 mg to 10g per patient per day which may be administered in single or multiple doses. In some embodiments the therapeutically effective amount is between a lower limit of 0.5 mg, 10 mg, 1 mg, 500.0 mg, 1000 mg, 1500 mg, 2000 mg, 2500 mg, 3000 mg, 3500 mg, 4000 mg, 4500 mg, 5000 mg, 5500 mg, 6000 mg, 6500 mg, 7000 mg, 7500 mg, 8000 mg, 8500 mg, 9000 mg, 9500 mg, and 10000 mg; and an upper limit of 10000 mg, 9500 mg, 9000 mg, 8500 mg, 8000 mg, 7500 mg, 7000 mg, 6500 mg, 6000 mg, 5500 mg, 5000 mg, 4500 mg, 4000 mg, 3500 mg, 3000 mg, 2500 mg, 2000 mg, 1500 mg, 1000 mg, 500.0 mg, 100 mg, 10 mg and 0.5 mg. In some embodiments, the therapeutically effective amount will be about 0.5 mg to 2500 mg per patient per day; in some embodiments about 0.5 mg to 200 mg per patient per day; in some embodiments about 0.5 mg to 500 mg
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SUBSTITUTE SHEET ( RULE 26 ) per patient per day; in some embodiments about 0.5 mg to 1000 mg per patient per day; and in yet some other embodiments about 5 mg to 50 mg per patient per day. Pharmaceutical compositions of the present invention may be provided in a solid dosage formulation such as comprising about 0.5 mg to 500 mg active ingredient, or comprising about 1 mg to 250 mg active ingredient. The pharmaceutical composition may be provided in a solid dosage formulation comprising for example about 1 mg, 5 mg, 10 mg, 25 mg, 50 mg, 100 mg, 200 mg, 250 mg, 500 mg or 1000 mg of active ingredient. For oral administration, the pharmaceutical compositions may be provided in the form of tablets containing 1.0 to 1000 milligrams of the active ingredient, such as 1, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900, 1000 and 2000 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. The compounds may be administered on a regimen of 1 to 4 times per day, such as once, twice, three times or four times per day.
DEFINITIONS
[0297] Molecular terms, when used in this application, have their common meaning unless otherwise specified.
10298] The articles "a" and "an" as used herein mean "one or more" or "at least one," unless otherwise indicated. That is, reference to any element of the present invention by the indefinite article "a" or "an" does not exclude the possibility that more than one of the element is present.
[0299] The term “acylamino” denotes a nitrogen radical adjacent to an acyl group.
[0300] As used herein, the term "alkyl" is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. For example, "C1-C17- alkyl" or "C1.17 alkyl" (or alkylene), is intended to include Ci, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, and C17 alkyl groups. Additionally, for example, "Ci-Ce alkyl" or "Ci-6 alkyl" denotes alkyl having 1 to 6 carbon atoms. Alkyl group can be unsubstituted or substituted with at least one hydrogen being replaced by another chemical group. In some embodiments the one of more hydrogen atoms is replaced by a chemical group selected from hydroxyl, and dimethylamino. Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, t-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl). Examples of substituted alkyl includes, but is not limited to, -CH2N(CH3)2, -CH2CH2N(CH3)2, and -CH2CH2CH2N(CH3)2.
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SUBSTITUTE SHEET ( RULE 26 ) [03011 "Alkenyl" is intended to include hydrocarbon chains of either straight or branched configuration having the specified number of carbon atoms and one or more, preferably one to three, carbon-carbon double bonds that may occur in any stable point along the chain. For example, "C2-C6 alkenyl" or "C2-6 alkenyl" (or alkenylene), is intended to include C2, C3, C4, C5, and G> alkenyl groups. The term "C2-17 alkenyl", is intended to include C2, C3, C4, C5, Ce, C7, Cs, C9, C10, Cn, C12, C13, C14, C15, Ci6, and C17 alkenyl groups. Examples of alkenyl include, but are not limited to, ethenyl, 1 -propenyl, 2-propenyl, 2- butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5- hexenyl, 2-methyl-2 -propenyl, and 4-methyl- 3 -pentenyl.
[0302] “Alkynyl" is intended to include hydrocarbon chains of either straight or branched configuration having one or more, preferably one to three, carbon-carbon triple bonds that may occur in any stable point along the chain. For example, "C2-C6 alkynyl" is intended to include C2, C3, C4, C5, and C<, alkynyl groups; such as ethynyl, propynyl, butynyl, pentynyl, and hexynyl.
[03031 The term "alkoxy" or "alkyloxy" refers to an -O-alkyl group. "Ci-Ce alkoxy" or "Ci-6 alkoxy" (or alkyloxy), is intended to include Ci, C2, C3, C4, C5, and Ce, alkoxy groups. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), and t-butoxy.
[0304] "Aryl" groups refer to monocyclic or polycyclic aromatic hydrocarbons, including, for example, thiazolyl, phenyl, and naphthyl. "Cs-Cio aryl" or " Ce-io aryl" refers to phenyl and naphthyl. Unless otherwise specified, "aryl", " Cg-Cw aryl," " Ce-io aryl," or "aromatic residue" may be unsubstituted or substituted with 1 to 5 groups selected from -OH, -OCH3, -CI, -F, -Br, -I, -CN, -NO2, -NH2, -NH(CH3), -N(CH3)2, -CF3, -OCF3, -C(O)CH3, - SCH3, -S(O)CH3, -S(O)2CH3, -CH3, -CH2CH3, -CO2H, and -CO2CH3.
[0305] The term "benzyl," as used herein, refers to a methyl group on which one of the hydrogen atoms is replaced by a phenyl group, wherein said phenyl group may optionally be substituted by one to five, preferably one to three, substituents independently selected from methyl, trifluoromethyl (-CF3), hydroxyl (-OH), methoxy (-OCH3), halogen, cyano (-CN), nitro (-NO2), -CO2Me, -CO2Et, and -CO2H. Representative examples of benzyl group include, but are not limited to, PI1CH2-, 4-MeO-C6H4CH2-, 2,4,6-tri-methyl-C6H2CH2-, and 3,4-di-Cl-C5H3CH2-.
[0306] The term “carboxyamido” denotes a carbonyl radical adjacent to an amino group.
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SUBSTITUTE SHEET ( RULE 26 ) [0307| A "compound," as used herein, refers to any type of substance or agent that is commonly considered a drug, or a candidate for use as a drug, as well as combinations and mixtures of the above. When referring to a compound of the invention, and unless otherwise specified, the term "compound" is intended to encompass not only the specified molecular entity but also its pharmaceutically acceptable, pharmacologically active analogs, including, but not limited to, salts, polymorphs, esters, amides, prodrugs, adducts, conjugates, active metabolites, and the like, where such modifications to the molecular entity are appropriate.
[0308] A “conventional mitochondrial uncoupler” as used herein describes a mitochondrial uncoupler that has properties that increase OCR and decreased MMP, and the concentrations for increasing OCR and dissipating MMP correlate.
[0309] As used herein, a "derivative" of a compound refers to a chemical compound that may be produced from another compound of similar structure in one or more steps. Non-limiting examples include replacement of H by an alkyl, acyl, or amino group.
[0310| As used herein, an "effective amount" or "therapeutically effective amount" means an amount sufficient to produce a selected effect, such as alleviating symptoms of a disease or disorder. In the context of administering compounds in the form of a combination, such as multiple compounds, the amount of each compound, when administered in combination with another compound(s), may be different from when that compound is administered alone. Thus, an effective amount of a combination of compounds refers collectively to the combination as a whole, although the actual amounts of each compound may vary. The term "more effective" means that the selected effect is alleviated to a greater extent by one treatment relative to the second treatment to which it is being compared.
]0311] The terms "formula" and "structure" are used interchangeably herein.
|0312] The term "halo" or "halogen" refers to fluoro, chloro, bromo, and iodo. "Haloalkyl" is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, substituted with one or more halogens. Examples of haloalkyl include, but are not limited to, fluoromethyl, difluoromethyl, trifluorom ethyl, and tri chloromethyl.
|0313] As used herein, the term "heteroaryl" is intended to mean stable monocyclic and polycyclic aromatic hydrocarbons that include at least one heteroatom ring member, such as sulfur, oxygen, or nitrogen. Heteroaryl groups include, without limitation, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, furyl, quinolyl, isoquinolyl, thienyl,
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SUBSTITUTE SHEET ( RULE 26 ) imidazolyl, thiazolyl, indolyl, pyrroyl, oxazolyl, benzofuryl, benzothienyl, benzthiazolyl, isoxazolyl, pyrazolyl, triazolyl, tetrazolyl, indazolyl, 1,2,4-thiadiazolyl, isothiazolyl, purinyl, carbazolyl, benzimidazolyl, indolinyl, benzodioxolanyl, and benzodioxane. Unless otherwise specified, heteroaryl groups may be unsubstituted or substituted with 1 to 5 groups selected from -OH, -OCH3, -CI, -F, -Br, -I, -CN, -NO2, -NH2, -NH(CH3), -N(CH3)2, -CF3, -OCF3, - C(O)CH3, -SCH3, -S(O)CH3, -S(O)2CH3, -CH3, -CH2CH3, -CO2H, and -CO2CH3 The nitrogen atom is substituted or unsubstituted (i.e., N or NR wherein R is H or another substituent, if defined). The nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., N— >0 and S(O)P, wherein p is 0, 1 or 2).
[0314| The term “heterocyclyl,” “heterocyclic” or “heterocyclyl ring” is defined as a saturated or partially unsaturated ring containing one to four hetero atoms or hetero groups selected from O, N, NH, -N(RZ)-, -S(O)- or -S(O)2- , wherein Rz is selected from alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, optionally substituted heterocyclyl, in a single or fused heterocyclic ring system having from three to twelve ring members. In a preferred embodiment, a heterocyclyl is a ring system having three to seven ring members Examples of a heterocyclyl group include, without limitation, azetindinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, morpholinyl, dioxothiomorphylinyl, tetrahydrofuranyl and azabicyclo[3.2.1]octanyl. Unless otherwise specified, heteroaryl groups may be unsubstituted or substituted with at least one groups selected from oxo, cyano, hydroxyl, alkoxy, -acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl.
[0315] The term “infectious disease” as described herein referes to a bacterial infection, or viral infection. Infectious diseases do not include infections caused by parasitic organism. In some embodiments the viral infection is an envelope virus. Examples of envelope viruses include SARS-CoV-2 , corana viruses and Ebola viruses.
[0316] The term “mitochondria-related condition or disorder” is defined as pathologic conditions caused by mitochondrial malfunction as reviewed and summarized in: A Mitochondrial Paradigm of Metabolic and Degenerative Diseases, Aging, and Cancer: A Dawn for Evolutionary Medicine, Annu Rev Genet. 2005; 39: 359, The rise of mitochondria in medicine, Mitochondrion 2016, 30:105-16, and Is Mitochondrial Dysfunction a Common Root of Noncommuni cable Chronic Diseases? Endocrine Reviews 2020,41(491-517), all of which are incorporated herein by reference. These conditions include, but are not limited to: genetic mitochondrial diseases, various types of cancer, autisim, neurodegenerative diseases, neuromuscular diseases, immunological diseases, metabolic diseases, aging, and aging- related noncommuni cable chronic diseases.
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SUBSTITUTE SHEET ( RULE 26 ) [0317| An “MMP -retaining compound” or a “mitochondrial membrane potentialretaining compound” is defined as a mitochondrial uncoupler that effectively increases OCR without significantly reducing MMP.
[0318] The term, "mitochondrial uncoupling", also referred to as "uncoupling", refers to the process whereby protons enter the mitochondrial matrix via a pathway independent of ATP synthase and thereby uncouple nutrient oxidation from ATP production. This process can be pharmacologically induced by small molecule mitochondrial protonophores, which directly shuttle protons across the mitochondrial inner membrane into the matrix. The primary pathway for energy production in aerobic cells involves the oxidation of nutrients (including fats, carbohydrates, and amino acids) in mitochondria, which promotes the efflux of protons out of the mitochondrial matrix. This process creates a pH and electrochemical gradient across the mitochondrial inner membrane. Protons normally re-enter the mitochondrial matrix via ATP synthase, which results in ATP production. Protons can also re-enter the mitochondrial matrix via pathways independent of ATP synthase, which 'uncouples' nutrient oxidation and proton efflux from ATP production.
[0319] The phrase " opthalmically acceptable" is employed herein to refer to those compounds, materials, pharmaceutical compositions, and/or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the eyes of human beings and animals without excessive toxicity, irritation, allergic response, and/or other problem or complication, commensurate with a reasonable benefit/risk ratio.
[0320] The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, pharmaceutical compositions, and/or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, and/or other problem or complication, commensurate with a reasonable benefit/risk ratio.
|0321] As used herein, "pharmaceutically acceptable salts" refer to derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic groups such as amines; and alkali or organic salts of acidic groups such as carboxylic acids. The pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, and nitric; and the salts prepared from organic
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SUBSTITUTE SHEET ( RULE 26 ) acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, and isethionic.
[0322] The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound that contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Company, Easton, PA, 1990, the disclosure of which is hereby incorporated by reference.
[0323] As used herein, the term "pharmaceutically acceptable carrier" includes any of the standard pharmaceutical carriers, such as a phosphate buffered saline solution, water, emulsions such as an oil/water or water/oil emulsion, and various types of wetting agents. The term also encompasses any of the agents approved by a regulatory agency of the US Federal government or listed in the US Pharmacopeia for use in animals, including humans.
[0324] A "prodrug" refers to an agent that is converted into the parent drug in vivo. Prodrugs are often useful because, in some situations, they may be easier to administer than the parent drug. They may, for instance, be bioavailable by oral administration whereas the parent is not. The prodrug may also have improved solubility in pharmaceutical compositions over the parent drug, or may demonstrate increased palatability, or be easier to formulate.
[0325] The terms "subject," “individual” or “patient” are used interchangeably and as used herein are intended to include human and non-human animals. Non-human animals includes all vertebrates, e g. mammals and non-mammals, such as non-human primates, sheep, dogs, cats, cows, horses, chickens, amphibians, and reptiles, although mammals are preferred, such as non-human primates, sheep, dogs, cats, cows and horses. Preferred subjects include human patients in need of enhancement of an immune response. The methods are particularly suitable for treating human patients having a disease or disorder described herein.
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SUBSTITUTE SHEET ( RULE 26 ) [0326| The terms “administration of’ and or “administering a” compound should be understood to mean providing a compound described herein or a prodrug thereof to the individual in need of treatment.
|0327] The term “treating” or “treatment” as used herein refers to administration of a compound or agent to a subject who has a disorder or is at risk of developing the disorder with the purpose to cure, alleviate, relieve, remedy, delay the onset of, prevent, or ameliorate the disorder, the symptom of the disorder, the disease state secondary to the disorder, or the predisposition toward the disorder.
EXAMPLES
103281 Processes for preparing compounds of the present invention, such as Formulas A, I, la, II, Ila, in and Illa, or for preparing intermediates useful for preparing compounds of Formulas A, I, la, II, Ha, III and Illa or other formulas of the present disclosure are provided as further embodiments of the invention or are known in the art. While the following text may exemplify specific compounds and corresponding routes of synthesis, it is not intended to limit the scope of the invention to such particular reference or examples. Various modifications may be made by those skilled in the art, in view of practical and economic considerations, such as the source of the agents and specific conditions of reactions.
I0329 | Chemical Abbreviations:
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SUBSTITUTE SHEET ( RULE 26 )
Example 1 -chloro-2-hydroxy-3-((methoxymethoxy)methyl)-A-(6-(trifluoromethyl)benzo[6?]thiazol-2- yl)benzamide(l)
SUBSTITUTE SHEET ( RULE 26 )
[0330| To a stirred solution of methyl 5-chl oro-3 -(hydroxymethyl)-2- methoxybenzoate (106 mg, 0.46 mmol) in DCM (5 mL) was added DIPEA (240 pL, 1.38 mmol), MOMC1 (105 pL, 1.38 mmol) and followed by DMAP (3 mg, 0.023 mmol). The reaction mixture was stirred at rt overnight. After the reaction was completed, DCM and sat. Ammonia chloride solution were added. The organic layer was dried and concentrated, the residue was purified via silica gel column chromatography to give methyl 5-chloro-2- methoxy-3-((methoxymethoxy)methyl)benzoate (122 mg, 97%) as yellow oil. ’H NMR (300 MHz, Chloroform-d) 5 7.72 (d, J = 2.8 Hz, 1H), 7.58 (d, J = 2.9 Hz, 1H), 4.73 (s, 2H), 4.64 (s, 2H), 3.90 (s, 3H), 3.83 (s, 3H), 3.40 (s, 3H).
[0331] To a stirred solution of methyl 5-chloro-2-methoxy-3- ((methoxymethoxy)methyl)benzoate (122 mg, 0.445 mmol) in MeOH (5 mL) was added 2.2 mL IN KOH solution. The resulting mixture was stirred at 60 °C overnight. After the mixture was cooled to rt, the reaction was partitioned between ethyl acetate and 2% citric acid. The ethyl acetate layer was washed with brine, dried over Na2SO4 and concentrated in vacuo. To this residue was added HBTU (98 mg, 0.258 mmol), DMF (3 mL) and DIPEA (187 pL, 1.075 mmol). The mixture was stirred for 10 minutes and then 6- (trifluoromethyl)benzo[d]thiazol-2-amine (47 mg, 0.215 mmol) was added. The resulting reaction was heated at 120 °C for 24 h. the mixture was cooled to rt, then separated between ethyl acetate and water. The organic layer was washed with brine, dried over NazSCL and concentrated in vacuo. Purification by column chromatography gave the 5-chloro-2-hydroxy- 3-((methoxymethoxy)methyl)-A-(6-(trifluoromethyl)benzo[<7]thiazol-2 yl)benzamide as a yellow solid (37 mg, 39%). XHNMR (400 MHz, Chloroform-d) 8 8.16 (s, 1H), 7.99 (d, J = 2.5 Hz, 1H), 7.91 (d, J = 6.1 Hz, 1H), 7.72 (d, J = 6.1 Hz, 1H), 7.48 (d, J = 2.5 Hz, 1H), 4.80 (s, 2H), 4.78 (s, 2H), 3.47 (s, 3H). MS (ESI) [M+Na]+ requires m/z 469.02, found m/z 468.55.
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SUBSTITUTE SHEET ( RULE 26 ) Example 2
5-chloro-2 -hydroxy-3 -((2 -methoxy ethoxy )methyl)-JV-(6-(trifluoromethyl)benzo[ ]thiazol-2- yl)benzamide (2)
[0332] To a stirred solution of 5-chloro-2-methoxybenzoic acid (5.59g, 30 mmol) in sulfuric acid (10.2 mL) and TFA (20.4 mL) at rt was added NBS (5.87g, 33 mmol). The pale solution was stirred at rt overnight. The resulting pale suspension was carefully poured onto crushed ice. The mixture was extracted with ethyl acetate. The ethyl acetate layer was dried over Na2SO4 and concentrated under reduced pressure. The light yellow residue was suspended in a minimum amount of DCM. The solid was collected, washed with cold DCM and dried under vacuum to yield 3 -bromo-5-chloro-2-m ethoxybenzoic acid as a white solid (8.00g, 100%). ‘H NMR (300 MHz, acetone) 6 7.86 (d, 1H, J=3.0Hz), 7.78(d, 1H, J=3.0Hz), 3.91(s, 3H).
[0333] To a stirred solution of 3 -bromo-5-chloro-2 -methoxybenzoic acid (6g, 22.6 mmol) in DMF (30 mL) was added potassium carbonate (31g, 226 mmol) followed by CH3I
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SUBSTITUTE SHEET ( RULE 26 ) (1.4 mL, 22.6 mmol). The mixture was stirred at rt for 24 h. Water was added and extracted with ethyl acetate two times. The combined organic layer was washed with water and brine and dried over sodium sulfate The organic layer was filtered and the solvent removed in vacuo to yield a pale yellow oil (6.18g, 97%).
[0334] A mixture of methyl 3-bromo-5-chloro-2-methoxybenzoate (756 mg, 2.7 mmol), methylboronic acid (324 mg, 5.4 mmol), Pd(OAc)2 (24 mg, 0.11 mmol), tricyclohexylphosphine (68 mg, 0.24 mmol) and potassium phosphate tribasic (1.9g, 8.96 mmol) was refluxed in toluene(10 mL) and water (1 mL) under N2 overnight. After the reaction was cooled, saturated NH4CI solution was added and extracted with ethyl acetate two times. The combined ethyl acetate layer was dried over NajSCh and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give methyl 5 -chi oro-2-m ethoxy-3 -methylbenzoate (520 mg, 91%) as a yellow oil. 'H NMR (300 MHz, chloroform) 5 7.62 (d, 1H, J=3.0Hz), 7.32(d, 1H, J=3.0Hz), 3.92(s, 3H), 3.82 (s, 3H), 2.30 (s, 3H).
[0335| To a flame dried flask was added NBS (183 mg, 1.03 mmol), AIBN (15 mg, 0.093 mmol) and a solution of methyl 5-chloro-2-methoxy-3-methylbenzoate (201g, 0.93 mmol) in CCI4 (10 mL). The suspension was refluxed in the dark overnight. The mixture was cooled to rt and concentrated. The residue was purified via silica gel column chromatography to give methyl 5-chloro-3-(bromomethyl)-2-methoxybenzoate as a colorless oil (236 mg, 87%). 1H NMR (300 MHz, cdcl3) 5 7.76 (d, J = 2.7 Hz, 1H), 7.54 (d, J = 2.7 Hz, 1H), 4.51 (s, 2H), 3.96 (s, 3H), 3.94 (s, 3H).
|0336] To a stirred solution of methyl 5-chl oro-3 -(bromomethyl)-2- methoxybenzoate (236 mg, 0.805 mmol) in 2-methoxyethanol (10 mL) was added 2NNaOH solution (7 mL). The resulting mixture was stirred at 75°C overnight and then concentrated in vacuo. The residue was dissolved in ethyl acetate and the resulting solution was washed with 2N HC1 dried over sodium sulfate and concentrated in vacuo. The residure was triturated with ether to give 5-chloro-2-methoxy-3-((2-methoxyethoxy)methyl)benzoic acid (204 mg, 93%) as yellow oil. XHNMR (300 MHz, cdcl3) 5 10.02 (brs, 1H), 7.86 (d, J = 2.8 Hz, 1H), 7.66 (d, J = 1.4 Hz, 1H), 4.62 (s, 2H), 3.87 (s, 3H), 3.75 - 3.67 (m, 2H), 3.66 - 3.56 (m, 2H), 3.41 (s, 3H). MS (ESI) [M+Na]+requires m/z 297.05, found m/z 296.6.
|0337] 5 -chloro-2-methoxy-3-((2 -methoxyethoxy )methyl)benzoic acid (90 mg,
0.328 mmol) was dissolved in DMF (3 mL). HBTU (149 mg, 0.394 mmol) was added followed by DIPEA (286 pL, 1.64 mmol). The resulting mixture was stirred at rt for 15mins, then 6-(trifluoromethyl)benzo[r/]thiazol-2-amine (72 mg, 0.328 mmol) was added. The
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SUBSTITUTE SHEET ( RULE 26 ) resulting mixture was stirred at 130 °C for 24h. Saturated NH4CI solution was added and extracted with ethyl acetate two times. The combined ethyl acetate layer was dried over NazSCri and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give 5-chloro-2-hydroxy-3-((2-methoxyethoxy)methyl)-A-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide as a yellow powder (34 mg, 30%). 'H NMR (300 MHz, acetone) 6 8 37 (s, 1H), 8.02 (s, 1H), 7.81 (d, J = 8.3 Hz, 1H), 7.69 (d, J = 8.4 Hz, 1H), 7.41 (s, 1H), 4.91 (s, 2H), 3.87 (s, 3H), 3.53 (brs, 2H), 2.94 (brs, 2H). MS (ESI) [M+Na]+requires m/z 483.04, found m/z 483.1.
Example 3
5-chloro-2-hydroxy-3-(((2 -methoxy ethyl)(methyl)amino)methyl)-A-(6- (trifluoromethyl)benzo[</]thiazol-2-yl)benzamide (3) and 5-chloro-2-hydroxy-3-(((2- methoxyethyl)(methyl)amino)methyl)-JV-(6-(trifluoromethyl)benzo[tZ]thiazol-2-yl)benzamide hydrochloride (3 A)
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SUBSTITUTE SHEET ( RULE 26 )
[0338] To a stirred solution of 5-chloro-2-methoxybenzoic acid (5.59g, 30 mmol) in sulfuric acid (10.2 mL) and TFA (20.4 mL) at rt was added NBS (5.87g, 33 mmol). The pale solution was stirred at rt overnight. The resulting pale suspension was carefully poured onto crushed ice. The mixture was extracted with ethyl acetate. The ethyl acetate layer dried over NazSCh and concentrated under reduced pressure. The light yellow residue was suspend in minimum amount of DCM. The solid was collected, washed with cold DCM and dried under vacuum to yield the 3-bromo-5-chloro-2-methoxybenzoic acid as a white solid (8.00g, 100%). 'l l NMR (300 MHz, acetone) S 7.86 (d, 1H, J=3.0Hz), 7.78(d, 1H, J=3.0Hz), 3.91(s, 3H).
[0339] To a stirred solution of 3 -bromo-5-chloro-2 -methoxybenzoic acid (6g, 22.6 mmol) in DMF (30 mL) was added potassium carbonate (31g, 226 mmol) followed by CH3I (1.4 mL, 22.6 mmol). The mixture was stirred at rt for 24 h. Water was added and extracted with ethyl acetate two times. The combined organic layer was washed with water and brine and dried over sodium sulfate. The organic layer was filtered and the solvent removed in vacuo to yield a pale yellow oil (6.18g, 97%).
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SUBSTITUTE SHEET ( RULE 26 ) [0340| A mixture of methyl 3-bromo-5-chloro-2-methoxybenzoate (756 mg, 2.7 mmol), methylboronic acid (324 mg, 5.4 mmol), Pd(OAc)2 (24 mg, 0.11 mmol), tricyclohexylphosphine (68 mg, 0 24 mmol) and potassium phosphate tribasic (1.9g, 8.96 mmol) was refluxed in toluene(10 mL) and water (1 mL) under N2 overnight. After the reaction was cooled, saturated NH4CI solution was added and extracted with ethyl acetate two times. The combined ethyl acetate layer was dried over Na2SO4 and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give methyl 5 -chi oro-2-methoxy-3 -methylbenzoate (520 mg, 91%) as a yellow oil. 'H NMR (300 MHz, chloroform) 8 7.62 (d, 1H, J=3.0Hz), 7.32(d, 1H, J=3.0Hz), 3 92(s, 3H), 3.82 (s, 3H), 2.30 (s, 3H).
[0341[ To a flame dried flask was added NBS (492 mg, 2.767 mmol), AIBN (57 mg, 0.346 mmol) and a solution of methyl 5-chloro-2-methoxy-3-methylbenzoate (495 mg, 2.306 mmol) in CC14 (10 mL). The suspension was refluxed in the dark overnight. The mixture was cooled to rt and concentrated. The residue was purified via silica gel column chromatography to give methyl 5-chloro-3-(bromomethyl)-2-methoxybenzoate as a colorless oil (588 mg, 87%). 'H NMR (300 MHz, cdcl3) 6 7.76 (d, J = 2.7 Hz, 1H), 7.54 (d, J = 2.7 Hz, 1H), 4.51 (s, 2H), 3.96 (s, 3H), 3.94 (s, 3H).
[0342] At 0 °C, to a stirred solution of methyl 5-chl oro-3 -(bromomethyl)-2- methoxybenzoate (500 mg, 1.71 mmol) in THF (6 mL) was added 2-methoxy-JV- methylethan- 1 -amine (370 pL, 3.41 mmol). The mixture was stirred at rt for 16 h. After completion of the reaction, the mixture was partitioned between NaHCCh and ethyl acetate. The aqueous layer was further extracted with ethyl acetate two times. The combined organic layer was washed with brine and dried over sodium sulfate. The solvent was removed under reduce pressure to afford methyl 5-chloro-2-methoxy-3-(((2- methoxyethyl)(methyl)amino)methyl)benzoate (437 mg, 85%) as a yellow oil. XH NMR (300 MHz, Chloroform-d) 8 7.67 (s, 2H), 3.91 (s, 3H), 3.81 (s, 2H), 3.61 (s, 2H), 3.52 (t, J = 5.7 Hz, 2H), 3.34 (s, 3H), 2.64 (t, J = 5.7 Hz, 2H), 2.29 (s, 3H). MS (ESI) [M+H]+ requires m/z 302.12, found m/z 301.60.
[0343] To a stirred solution of methyl 5-chloro-2-methoxy-3-(((2- methoxyethyl)(methyl)amino)methyl)benzoate (352 mg, 1.17 mmol) in MeOH (5 mL) was added 5.0 mL IN KOH solution. The resulting mixture was stirred at 50 °C overnight. The solvent was evaporated out and 4N HC1 in dioxane (2 mL) was added to the residue. The mixture was stirred for another 10 minutes before it was concentrated and dried under vacumn. To this residue was added HBTU (532 mg, 1.404 mmol), DMF (5 mL) and DIPEA
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SUBSTITUTE SHEET ( RULE 26 ) (1.02 mL, 5.85 mmol). The mixture was stirred for 10 minutes and then 6- (trifluoromethyl)benzo[d]thiazol-2-amine (255 mg, 1.17 mmol) was added. The resulting reaction was heated at 130 °C for 24hs. After cooling to rt, the mixture was separated between ethyl acetate and water. The organic layer was washed with brine, dried over NazSCh and concentrated in vacuo. Purification by column chromatography gave 5-chloro-2- hydroxy-3-(((2-methoxyethyl)(methyl)amino)methyl)-Ar-(6-(trifluoromethyl)benzo[<7]thiazol- 2-yl)benzamide as a yellow solid (220 mg, 40%). 'H NMR (300 MHz, Methanol-d4) 8 8.24 (s, 1H), 7.93 (s, 1H), 7.84 (d, J = 8.4 Hz, 1H), 7.69 (d, J = 9.0 Hz, 1H), 7.31 (s, 1H), 4.31 (s, 2H), 3.78 (t, J=6.0Hz, 2H), 3.44 (s, 3H), 3.35 (t, J=6.0Hz, 2H), 2 85 (s, 3H). MS (ESI) [M+H]+ requires m/z 474.09, found m/z 473.55.
[0344 To a stirred solution of 5-chloro-2-hydroxy-3-(((2- methoxyethyl)(methyl)amino)methyl)-A-(6-(trifluoromethyl)benzo[</]thiazol-2-yl)benzamide (126 mg, 0.266 mmol) in THF (5 mL) was added 4. ON HC1 in dioxane (70 pL, 0.266 mmol). The mixture was stirred at rt for 20mins. The solvent was removed under reduce pressure and the resulting residue was washed with diethyl ether to afford 5-chloro-2-hydroxy-3-(((2- methoxyethyl)(methyl)amino)methyl)-A-(6-(trifluoromethyl)benzo[</]thiazol-2-yl)benzamide hydrochloride as a yellow solid (135 mg, 100%).
Example 4
5-chloro-2V-(2-chloro-4-(trifluoromethyl)phenyl)-2-hydroxy-3-((2- m ethoxy ethoxy )methyl)benzamide (4)
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SUBSTITUTE SHEET ( RULE 26 )
[0345] To a stirred solution of 5 -chi oro-2-methoxy-3 -methylbenzoic acid (1.80g, 8.99 mmol) in DMF (10 mL) was added potassium carbonate (12.4g, 89.9 mmol) followed by CH3I (0.56 mL, 8.99 mmol). The mixture was stirred at rt for 24 h. Water was added and extracted with ethyl acetate two times. The combined organic layer was washed with water and brine and dried over sodium sulfate. The organic layer was filtered and the solvent removed in vacuo to yield methyl 5-chloro-2-methoxy-3-methylbenzoate as a pale yellow oil (1.13g, 63%). 'HNMR (300 MHz, cdch) S 7.52 (d, J = 2.7 Hz, 1H), 7.22 (d, J = 2.7 Hz, 1H), 3.83 (s, 3H), 3.74 (s, 3H), 2.21 (s, 3H).
[0346] At -78 °C, to a stirred solution of methyl 5-chloro-2-methoxy-3- methylbenzoate (241 mg, 1.12 mmol) in anhydrous DCM (5 mL) was added BBn (1.0M in DCM, 2.25 mL) dropwise. After addition, the reaction was allowed to warm to rt slowly and the mixture was stirred at rt for 2h. After completion of the reaction, the reaction mixture was cooled in an ice bath and MeOH and water were added to quench the reaction. The mixture was separated between DCM and water. The organic layer was washed with water and brine, dried over sodium sulfate and concentrated in vacuo to afford methyl 5-chl oro-2 -hy droxy-3 - methylbenzoate as pale yellow solid (220 mg, 98%), which was used in the next step without further purification.
10347] To a stirred solution of methyl 5-chl oro-2-hy droxy-3 -methylbenzoate (220 mg, 1.1 mmol) in anhydrous DCM (5 mL) was added pyridine (443 pL, 5.5 mmol), (Boc)2O (504 mg, 2.30 mmol) and DMAP (13 mg, 0.11 mmol). The resulting mixture was stirred at rt
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SUBSTITUTE SHEET ( RULE 26 ) for 2 days. After completion of the reaction, solvent was evaporated and the resulting residue was purified via silica gel column chromatography to yield methyl 2-((tert- butoxycarbonyl)oxy)-5-chloro-3-methylbenzoate (290 mg, 88%) as a colorless oil. 'H NAIR (300 MHz, cdch) 5 7.80 (dd, J = 2.7, 0.6 Hz, 1H), 7.40 (dd, J = 2.7, 0.7 Hz, 1H), 3.89 (s, 3H), 2.26 (s, 3H), 1.58 (s, 9H).
103481 To a flame dried flask was added NBS (205 mg, 1.156 mmol), AIBN (24 mg, 0.145 mmol) and a solution of methyl 2-((/er/-butoxycarbonyl)oxy)-5-chloro-3- methylbenzoate (289 mg, 0.963 mmol) in CCL (5 mL). The suspension was refluxed in the dark overnight. The mixture was cooled to rt and concentrated. The residue was purified via silica gel column chromatography to give methyl 3-(bromomethyl)-2-((/er/- butoxycarbonyl)oxy)-5-chlorobenzoate as a colorless oil (220 mg, 62%). 'H MR (300 MHz, cdch) 5 7.95 (d, J = 2.2 Hz, 1H), 7.61 (d, J = 2.3 Hz, 1H), 4.45 (s, 2H), 3.91 (s, 3H), 1.58 (s, 9H).
[0349] To a stirred solution of methyl 3-(bromomcthyl)-2-((/e/7- butoxycarbonyl)oxy)-5-chlorobenzoate (220 mg, 0.582 mmol) in 2-methoxyethanol (10 mL) was added 2N NaOH solution (5 mL). The resulting mixture was stirred at rt overnight and then concentrated in vacuo. The residue was dissolved in ethyl acetate and the resulting solution was washed with 2N HC1 dried over sodium sulfate and concentrated in vacuo. The residue was triturated with ether to give 5-chloro-2-hydroxy-3-((2- methoxyethoxy)methyl)benzoic acid (145 mg, 92%) as a white oil. 'H NMR (300 MHz, cdch) 5 10.76 (s, 2H), 7.67 (d, J = 2.6 Hz, 1H), 7.59 (d, J = 2.5 Hz, 1H), 4.61 (s, 2H), 3.79 (dd, J = 5.9, 2.7 Hz, 2H), 3.72 (dd, J = 5.9, 2.8 Hz, 2H), 3.49 (s, 3H). MS (ESI) [M+Na]+ requires m/z 283.03, found m/z 282.55.
[0350| 5-chl oro-2 -hydroxy-3 -((2-methoxyethoxy)methyl)benzoic acid (65 mg,
0.249 mmol) was dissolved in THF (3.0 mL), followed by the addition of a catalytic amount of DMF (1 drop) and oxalyl chloride (26 pL, 0.299 mmol) respectively. The reaction was allowed to stir at rt for 30 min and was concentrated in vacuo. The residue was re-dissolved in dioxane(5.0 mL) and 2-chloro-4-(trifluoromethyl)aniline (35 pL, 0.25 mmol) was added. The mixture was refluxed overnight. Solvent was evaporated and the resulting residue was purified via silica gel column chromatography to yield -chloro-A-(2-chloro-4- (trifluoromethyl)phenyl)-2-hydroxy-3 -((2 -m ethoxy ethoxy)methyl)benzamide (35 mg, 35%) as a white solid. LH NMR (500 MHz, cdch) 5 10.25 (s, 1H), 10.20 (s, 1H), 8.76 (d, J = 8.6 Hz, 1H), 7.96 (d, J = 2.6 Hz, 1H), 7.69 (d, J = 1.7 Hz, 1H), 7.58 (dd, J = 8.8, 2.1 Hz, 1H),
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SUBSTITUTE SHEET ( RULE 26 ) 7.36 (d, J = 2.6 Hz, 1H), 4.73 (s, 2H), 3.82 - 3.76 (m, 2H), 3.66 - 3.62 (m, 2H), 3.45 (s, 3H).
MS (ESI) [M+Na]+ requires m/z 460.03, found m/z 459.95.
Example 5
5-chloro-2-hydroxy-3-((2 -methoxy ethoxy )methyl)-iV-(6-(tri fluoromethoxy )benzo[c/]thiazol- 2-yl)benzamide (5)
|0351] 5-chloro-2-hydroxy-3-((2-methoxyethoxy)methyl)benzoic acid (77 mg, 0.28 mmol, Example 2) was dissolved in DCM (3.0 mL), followed by the addition of a catalytic amount of DMF (1 drop) and oxalyl chloride (40 pL, 0.34 mmol) respectively. The reaction was allowed to stir at rt for 30 minutes and concentrated in vacuo. The residue was re-dissolved in THF (5.0 mL), and Hunig’s base (59 pL, 0.34 mmol) and 6- (trifluoromethoxy)benzo[t/]thiazol-2-amine (65 mg, 0.28 mmol) were added. The mixture was stirred at rt for 48 hours before silica gel was added to quench the reaction. Solvent was evaporated and the resulting residue was purified via silica gel column chromatography to yield 5-chloro-2-methoxy-3-((2-methoxyethoxy)methyl)-jV-(6-
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SUBSTITUTE SHEET ( RULE 26 ) (trifluoromethoxy )benzo[ ]thiazol-2-yl)benzamide (27 mg, 34% yield) as a white solid. 'H NMR (500 MHz, cdch) 5 8.13 (d, J = 2.8 Hz, 1H), 7.83 (d, J = 8.8 Hz, 1H), 7.73 (d, J = 2.8 Hz, 2H), 7.34 (ddd, J = 8.8, 2.4, 0.8 Hz, 1H), 4.66 (s, 2H), 3.99 (s, 3H), 3.75 - 3.72 (m, 2H), 3.62 (dd, J = 3.9, 2.5 Hz, 2H), 3.41 (s, 3H). MS (ESI) [M+H]+ requires m/z 491.07, found m/z 491.10.
[0352| A solution of 5-chloro-2-methoxy-3-((2-methoxyethoxy)methyl)-jV-(6- (trifluoromethoxy)benzo[d]thiazol-2-yl)benzamide (27 mg, 0.055 mmol,) in DMF (3 mL) is mixed with sodium ethoxide (18 mg, 0.275 mmol) and the resulting suspension is heated at 140 °C overnight. After completion of the reaction, IN HC1 was added to the reaction and extracted with ethyl acetate two times. The combined ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give 5-chloro-2-hydroxy-3-((2-methoxyethoxy)methyl)-/V-(6- (trifluoromethoxy)benzo[d]thiazol-2-yl)benzamide (13 mg, 50%) as a yellow solid. 'H NMR (500 MHz, cdch) 5 8.13 (d, J = 2.8 Hz, 1H), 7.83 (d, J = 8.8 Hz, 1H), 7.73 (d, J = 2.8 Hz, 2H), 7.34 (ddd, J = 8.8, 2.4, 0.8 Hz, 1H), 4.66 (s, 2H), 3.75 - 3.72 (m, 2H), 3.62 (dd, J = 3.9, 2.5 Hz, 2H), 3.41 (s, 3H). MS (ESI) [M+H]+ requires m/z 477.05, found m/z 477.30.
Example 6
5-chloro-2-hydroxy-3-((2 -methoxy ethoxy )methyl)-/V-(5-(trifluoromethyl)pyrazin-2- yl)benzamide (6)
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SUBSTITUTE SHEET ( RULE 26 )
[0353] 5-chloro-2-methoxy-3-((2 -methoxyethoxy )methyl)benzoic acid (90 mg,
0.33 mmol, Example 2) was dissolved in DCM (3.0 mL), followed by the addition of catalytic amount of DMF (10 pL) and oxalyl chloride (34 pL, 0.39 mmol) respectively. The reaction was allowed to stir at rt for 30 minutes, concentrated in vacuo, and the 5-chloro-2- methoxy-3-((2-methoxyethoxy)methyl)benzoyl chloride residue was re-dissolved in THF (3.0 mL). In another flask, 5-(trifluoromethyl)pyrazin-2-amine (54.0 mg, 0.33 mmol) was dissolved in THF (3.0 mL) followed by addition of NaH (16.0 mg, 0.439 mmol, 60% in mineral oil) The mixture was stirred for 10 minutes before it was added dropwise at rt to the flask containing the freshly prepared 5-chloro-2-methoxy-3-((2- methoxyethoxy)methyl)benzoyl chloride. The reaction was stirred at rt for 2h before silica gel was added to quench the reaction. Solvent was evaporated and the resulting residue was purified via silica gel column chromatography to yield 5-chloro-2-methoxy-3-((2- methoxyethoxy)methyl)-A-(5-(trifluoromethyl)pyrazin-2-yl)benzamide as a white solid (32.0 mg, 23% yield). ‘H NMR (300 MHz, cdch) 8 10.54 (s, 1H), 9.82 (d, J = 1.1 Hz, 1H), 8.67 (s, 1H), 8.11 (d, J = 2.8 Hz, 1H), 7.71 (d, J = 2.8 Hz, 1H), 4.68 (s, 2H), 3.95 (s, 3H), 3.79 - 3.71 (m, 2H), 3.67 - 3.59 (m, 2H), 3.43 (s, 3H). MS (ESI) [M+H]+ requires m/z 420.10, found m/z 420.30.
10354] A solution of 5-chloro-2-methoxy-3-((2-methoxyethoxy)methyl)-A-(5- (trifluoromethyl)pyrazin-2-yl)benzamide (32 mg, 0.076 mmol) in DMF (3 mL) is mixed with sodium ethanethiolate (32 mg, 0.382 mmol) and the resulting suspension is heated at 130 °C overnight. After completion of the reaction, IN HC1 was added to the reaction and extracted
SUBSTITUTE SHEET ( RULE 26 ) with ethyl acetate two times. The combined ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give 5-chloro-2-hydroxy-3-((2-methoxyethoxy)methyl)-A-(5- (trifluoromethyl)pyrazin-2-yl)benzamide (18 mg, 62%) as a yellow solid. XHNMR (300 MHz, acetone) 8 9.66 (s, 1H), 8.82 (s, 1H), 8.13 (s, 1H), 7.56 (s, 1H), 4.74 (s, 2H), 3.88 - 3.74 (m, 2H), 3.74 - 3.61 (m, 2H), 3.41 (s, 3H). MS (ESI) [M+H]+ requires m/z 406.08, found m/z 406.20.
Example 7
5-chloro-2-hydroxy-3-((2-hydroxy ethoxy )methyl)-A-(6-(trifluoromethyl)benzo|T ]thiazol-2- yl)benzamide (7)
[03551 A mixture of methyl 3-bromo-5-chloro-2-methoxybenzoate (756 mg, 2.7 mmol, Example 2), methylboronic acid (324 mg, 5.4 mmol), Pd(OAc)2 (24 mg, 0.11 mmol), tricyclohexylphosphine (68 mg, 0.24 mmol) and potassium phosphate tribasic (1.9g, 8.96
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SUBSTITUTE SHEET ( RULE 26 ) mmol) was refluxed in toluene(10 mL) and water (1 mL) under N2 overnight. After the reaction was cooled, saturated NH4CI solution was added and extracted with ethyl acetate two times. The combined ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give methyl 5-chloro-2-methoxy-3-methylbenzoate (520 mg, 91%) as a yellow oil. XH NMR (300 MHz, chloroform) 8 7.62 (d, 1H, J=3.0Hz), 7.32(d, 1H, J=3.0Hz), 3.92(s, 3H), 3.82 (s, 3H), 2.30 (s, 3H).
[0356] To a flame dried flask was added NBS (183 mg, 1.03 mmol), AIBN (15 mg, 0 093 mmol) and a solution of methyl 5-chloro-2-methoxy-3-methylbenzoate (201 mg, 0.93 mmol) in CCL (10 mL). The suspension was refluxed in the dark overnight. The mixture was cooled to rt and concentrated. The residue was purified via silica gel column chromatography to give methyl 5-chloro-3-(bromomethyl)-2-methoxybenzoate as a colorless oil (236 mg, 87%). 'H NMR (300 MHz, cdch) 8 7.76 (d, J = 2.7 Hz, 1H), 7.54 (d, J = 2.7 Hz, 1H), 4.51 (s, 2H), 3.96 (s, 3H), 3.94 (s, 3H).
[0357J To a stirred solution of methyl 5-chl oro-3 -(bromomethyl)-2- methoxybenzoate (222 mg, 0.76 mmol) in 2-(terZ-butoxy)ethan-l-ol (4 mL) was added 2N NaOH solution (2 mL). The resulting mixture was stirred at rt for 4h and then concentrated in vacuo. The residue was dissolved in ethyl acetate and the resulting solution was washed with 2N HC1 dried over sodium sulfate and concentrated in vacuo. The residure was triturated with ether to give 3-((2-(to7-butoxy)ethoxy)methyl)-5-chloro-2-methoxybenzoic acid as a yellow oil (100%). MS (ESI) [M-H]' requires m/z 315.10, found m/z 315.60.
|0358] To a stirred solution of 3-((2-(terLbutoxy)ethoxy)methyl)-5-chloro-2- methoxybenzoic acid (240 mg, 0.76 mmol) in DMF (5 mL) was added HBTU (345 mg, 0.91 mmol) and DIPEA (662 pL, 3.8 mmol). The mixture was stirred for 10 minutes and then 6- (trifluoromethyl)benzo[d]thiazol-2-amine (165 mg, 0.76 mmol) was added. The resulting reaction was heated at 120 °C for 24 h. After cooled to rt, the mixture was separated between ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and concentrated in vacuo. Purification by column chromatography gave 3-( 2-(tert- butoxy)ethoxy)methyl)-5-chloro-2-hydroxy-JV-(6-(trifluoromethyl)benzo[d]thiazol-2- yl)benzamide as a yellow solid (27 mg, 10%).1HNMR (300 MHz, acetone) 8 8.35 (s, 1H), 7.96 (s, 1H), 7.72 (dd, J= 19.4, 8.5 Hz, 2H), 7.41 (s, 1H), 4.89 (s, 2H), 4.06 (s, 2H), 3.82 (s, 2H), 1.15(s, 9H). MS (ESI) [M+H]+ requires m/z 503.10, found m/z 503.20.
[0359] To a stirred solution of 3-((2-(Ze/7-butoxy)ethoxy)methyl )-5-chloro-2- hydroxy-A-(6-(trifluoromethyl)benzo[ ]thiazol-2-yl)benzamide (27 mg, 0.054 mmol) in
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SUBSTITUTE SHEET ( RULE 26 ) DCM (3 mL) was added TFA (1 mL) and the mixture was stirred at rt overnight. The solvent was evaporated out and the remaining residue was subjected to flash column to afford 5- chloro-2-hydroxy-3-((2-hydroxyethoxy)methyl)-Ar-(6-(trifluoromethyl)benzo[<7]thiazol-2- yl)benzamide as a yellow solid (19 mg, 83%). XHNMR (300 MHz, acetone) 8 8.35 (s, 1H), 7.96 (s, 1H), 7.72 (dd, J= 19.4, 8.5 Hz, 2H), 7.41 (s, 1H), 4.89 (s, 2H), 4.06 (s, 2H), 3.82 (s, 2H). MS (ESI) [M+H]+ requires m/z 447.04, found m/z 447.10.
Example 8
5-chloro-jV-(6-fluorobenzo[tZ]thiazol-2-yl)-2-hydroxy-3-((2- m ethoxy ethoxy )methyl)benzamide (8)
SUBSTITUTE SHEET ( RULE 26 ) [0360] To a flame dried flask was added NBS (183 mg, 1.03 mmol), AIBN (15 mg, 0.093 mmol) and a solution of methyl 5-chloro-2-methoxy-3-methylbenzoate (201g, 0.93 mmol) in CC14 (10 mL). The suspension was refluxed in the dark overnight The mixture was cooled to rt and concentrated. The residue was purified via silica gel column chromatography to give methyl 3-(bromomethyl)-5-chloro-2 -methoxybenzoate as a colorless oil (236 mg, 87%). H NVIR (300 MHz, cdch) 87.76 (d, J = 2.7 Hz, 1H), 7.54 (d, J = 2.7 Hz, 1H), 4.51 (s, 2H), 3.96 (s, 3H), 3.94 (s, 3H).
10361] To a stirred solution of 3 -(bromomethyl)-5-chl oro-2 -methoxybenzoate (180 mg, 0.614 mmol) in 2-methoxyethanol (5 mL) was added 2N NaOH solution (4 mL). The resulting mixture was stirred at rt overnight and then concentrated in vacuo. The residue was dissolved in ethyl acetate and the resulting solution was washed with 2N HC1 dried over sodium sulfate and concentrated in vacuo. The residure was triturated with ether to give 5- chloro-2-methoxy-3-((2-methoxy ethoxy )methyl)benzoic acid (170 mg, 100%) as yellow oil. 'H NMR (300 MHz, cdch) 8 10.02 (brs, 1H), 7.86 (d, J = 2.8 Hz, 1H), 7.66 (d, J = 1.4 Hz, 1H), 4.62 (s, 2H), 3.87 (s, 3H), 3.75 - 3.67 (m, 2H), 3.66 - 3.56 (m, 2H), 3.41 (s, 3H). MS (ESI) [M+Na]+requires m/z 297.05, found m/z 296.6.
10362] 5-chloro-2-methoxy-3-((2 -methoxyethoxy )methyl)benzoic acid (85 mg,
0.31 mmol) was dissolved in DCM (3.0 mL), followed by the addition of a catalytic amount of DMF (1 drop) and oxalyl chloride (32 pL, 0.372 mmol) respectively. The reaction was allowed to stir at rt for 30 minutes and was concentrated in vacuo. The residue was redissolved in THF (5.0 mL), and Hunig’s base (162 pL, 0.93 mmol) and 6- fluorobenzo[<7]thiazol-2-amine (52 mg, 0.31 mmol) were added. The mixture was stirred at rt for 48 hours before silica gel was added to quench the reaction. Solvent was evaporated and the resulting residue was purified via silica gel column chromatography to yield 5-chloro-A- (6-fluorobenzo[<7]thiazol-2-yl)-2-methoxy-3-((2-methoxy ethoxy )methyl)benzamide (52 mg, 41%) as a white solid. MS (ESI) [M+H]+ requires m/z 425.08, found m/z 425.10.
[0363] A solution of 5-chloro-A-(6-fluorobenzo[t ]thiazol-2-yl)-2-methoxy-3-((2- methoxyethoxy)methyl)benzamide (52 mg, 0.123 mmol) in DMF (3 mL) was mixed with sodium ethanethiolate (52 mg, 0.613 mmol) and the resulting suspension was heated at 130 °C overnight. After completion of the reaction, IN HC1 was added to the reaction and extracted with ethyl acetate two times. The combined ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give 5-chloro-;V-(6-fluorobenzo[c/]thiazol-2-yl)-2-hydroxy-3- ((2-methoxy ethoxy )methyl)benzamide (20 mg, 40%) as a yellow solid. LH NMR (300 MHz,
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SUBSTITUTE SHEET ( RULE 26 ) acetone) 5 8.05 (s, 1H), 7.77 (dd, J= 8.4, 2.5 Hz, 1H), 7.61 (s, 1H), 7.47 (s, 1H), 7.20 (t, J = 8.8 Hz, 1H), 4.79 (s, 2H), 3.83 - 3.72 (m, 4H), 3.47 (s, 3H). MS (ESI) [M+H]+ requires m/z 411 06, found m/z 411 10.
Example 9
5-chloro-2-hydroxy-3-(methoxymethyl)-/V-(6-(trifluoromethyl)benzo[tZ]thiazol-2- yl)benzamide (9)
[0364] To a flame dried flask was added NBS (183 mg, 1.03 mmol), AIBN (15 mg, 0.093 mmol) and a solution of methyl 5-chloro-2-methoxy-3-methylbenzoate (201g, 0.93 mmol) in CC14 (10 mL). The suspension was refluxed in the dark overnight. The mixture was cooled to rt and concentrated. The residue was purified via silica gel column chromatography to give methyl 3-(bromomethyl)-5-chloro-2 -methoxybenzoate as a colorless oil (236 mg, 87%). 'H NMR (300 MHz, cdch) 57.76 (d, J = 2.7 Hz, 1H), 7.54 (d, J = 2.7 Hz, 1H), 4.51 (s,
2H), 3.96 (s, 3H), 3.94 (s, 3H).
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SUBSTITUTE SHEET ( RULE 26 ) [0365| To a stirred solution of methyl 3-(bromomethyl)-5-chloro-2- methoxybenzoate (270 mg, 0.921 mmol) in methanol (10 mL) was added 2NNaOH solution (7 mL). The resulting mixture was stirred at rt overnight and then concentrated in vacuo. The residue was dissolved in ethyl acetate and the resulting solution was washed with 2N HC1 dried over sodium sulfate and concentrated in vacuo. The residure was triturated with ether to give 5-chloro-2-methoxy-3-(methoxymethyl)benzoic acid (217 mg, 83%) as yellow oil which was used in the next step without further purification. MS (ESI) [M-H]' requires m/z 229.02, found m/z 229.40.
[0366] 5-chloro-2-methoxy-3-(methoxymethyl)benzoic acid (217 mg, 0.94 mmol) was dissolved in DCM (5.0 mL), followed by the addition of a catalytic amount of DMF (1 drop) and oxalyl chloride (97 pL, 1.13 mmol) respectively. The reaction was allowed to stir at rt for 30 min and concentrated in vacuo. The residue was re-dissolved in THF (5.0 mL), and Hunig’s base (197 pL, 1.13 mmol) and 6-(trifluoromethyl)benzo[tZ]thiazol-2-amine (206 mg, 0.94 mmol) were added. The mixture was stirred at rt for 48 hours before silica gel was added to quench the reaction. Solvent was evaporated and the resulting residue was purified via silica gel column chromatography to yield 5-chloro-2-methoxy-3-(methoxymethyl)-7V-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (165 mg, 45%) as a yellow solid. MS (ESI) [M+H]+ requires m/z 431.05, found m/z 431.10.
[0367] A solution of 5-chloro-2-methoxy-3-(methoxymethyl)-A-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (100 mg, 0.23 mmol) in DMF (5 mL) was mixed with sodium ethanethiolate (97 mg, 1.16 mmol) and the resulting suspension was heated at 130 °C overnight. After completion of the reaction, IN HC1 was added to the reaction and extracted with ethyl acetate two times. The combined ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give 5-chloro-2-hydroxy-3-(methoxymethyl)-7V-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (90 mg, 94%) as a white solid. 'H NMR (300 MHz, cdch) 8 8.16 (s, 1H), 8.13 (d, J= 2.6 Hz, 1H), 7.87 (d, J= 8.6 Hz, 1H), 7.73 (d, J = 8.9 Hz, 1H), 7.49 (d, J= 2.6 Hz, 1H), 4.67 (s, 2H), 3.55 (s, 3H). MS (ESI) [M-H]' requires m/z 415.02, found m/z 415.40.
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SUBSTITUTE SHEET ( RULE 26 ) Example 10
5-chloro-2-hydroxy-3-(pyri din-3-yl )-Af-(6-(tri fluoromethyl )benzo[<7]thiazol -2- yl)benzamide(lO) and 5-chloro-2-hydroxy-3-(pyridin-3-yl)-A-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide hydrochloride (10A)
[0368] A mixture of methyl 3-bromo-5-chloro-2-methoxybenzoate (280 mg, 1.0 mmol, Example 2), 3-pyridine boronic acid (184 mg, 1.5 mmol), PdC12(dpppf)-DCM(81 mg, 0.1 mmol) and 2M sodium carbonate (2 m ) in dioxane (5 m ) was heated to 75 °C overnight under nitrogen. After cooling, the reaction mixture was partitioned between water and ethyl acetate. The organic layer was washed with brine and dried over sodium sulfate. After concentration, the residue was purified via silica gel column chromatography to give methyl 5 -chi oro-2-m ethoxy-3 -(pyri din-3 -yl)benzoate (140 mg, 52%) as a white solid. 'H NMR (300 MHz, cdch) 5 8.74 (d, J= 40.1 Hz, 2H), 7.95 (d, J= 6.3 Hz, 1H), 7.81 (d, J= 1.5
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SUBSTITUTE SHEET ( RULE 26 ) Hz, 1H), 7.47 (brs, J= 17.7 Hz, 2H), 3.96 (s, 3H), 3.52 (s, 3H). MS (ESI) [M+H]+ requires m/z 278.06, found m/z 278.20.
|0369] To a stirred solution of methyl 5-chloro-2-methoxy-3-(pyridin-3- yl)benzoate (140 mg, 0.505 mmol) in MeOH (4 mL) was added 2.5 mL IN KOH solution. The resulting mixture was stirred at rt overnight. The solvent was evaporated out and 10% citric acid was added to the residue to pH=3. The mixture was extracted with DCM two times. The combined organic layer was dried over sodium sulfate and concentrated to afford 5-chloro-2-methoxy-3-(pyridin-3-yl)benzoic acid (132 mg, 100%) which was used directly in the next step without further purification as a yellow solid. MS (ESI) [M+H]+ requires m/z 264.04, found m/z 264.20.
[0370] 5-chloro-2-methoxy-3-(pyridin-3-yl)benzoic acid (132 mg, 0.50 mmol) was dissolved in DCM (3.0 mL) and THF (3.0 mL), followed by the addition of a catalytic amount of DMF (1 drop) and oxalyl chloride (52 pL, 0.60 mmol) respectively. The reaction was allowed to stir at rt for 30 minutes and then concentrated in vacuo. The residue was redissolved in THF (5.0 mL), and Hunig’s base (104 pL, 0.60 mmol) and 6- (trifluoromethyl)benzo[4/]thiazol-2-amine (109 mg, 0.50 mmol) were added. The mixture was stirred at rt for 48 hours before silica gel was added to quench the reaction. Solvent was evaporated and the resulting residue was purified via silica gel column chromatography to yield 5-chloro-2-methoxy-3-(pyridin-3-yl)-/V-(6-(trifluoromethyl)benzo[ ]thiazol-2- yl)benzamide (20 mg, 10%) as a yellow solid. 'H NMR (300 MHz, acetone) 5 8.88 (s, 1H), 8.59 (s, 1H), 8.40 (s, 1H), 8.22 (s, 1H), 8.11 (d, J= 7.7 Hz, 1H), 7.85 (dd, J= 17.8, 9.1 Hz, 2H), 7.63 (s, 1H), 7.60 - 7.46 (m, 1H), 3.28 (s, 3H). MS (ESI) [M+H]+ requires m/z 464.05, found m/z 464.20.
[0371| At -78 °C, To a stirred solution of 5-chloro-2-methoxy-3-(pyridin-3-yl)-A- (6-(trifluoromethyl)benzo[tZ]thiazol-2-yl)benzamide (20 mg, 0.043 mmol) in anhydrous DCM (5 mL) was added BBr, (1.0M in DCM, 129 pL) dropwise. After addition, the reaction was allowed to warm to rt slowly and the mixture was stirred at rt for 2h. After completion of the reaction, the reaction mixture was cooled in an ice bath and MeOH and water was added to quench the reaction. The mixture was separated between DCM and water. The organic layer was washed with water, brine and dried over sodium sulfate, concentrated in vacuo. The residue was purified via silica gel column chromatography to afford 5-chloro-2-hydroxy-3- (pyridin-3-yl)-A-(6-(trifluoromethyl)benzo[ ]thiazol-2-yl)benzamide (12 mg, 63%) as a yellow solid. H NMR (300 MHz, acetone) 5 8.88 (s, 1H), 8.59 (s, 1H), 8.40 (s, 1H), 8.22 (s,
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SUBSTITUTE SHEET ( RULE 26 ) 1H), 8.11 (d, J= 7.7 Hz, 1H), 7.85 (dd, J= 17.8, 9.1 Hz, 2H), 7.63 (s, 1H), 7.60 - 7.46 (m, 1H). MS (ESI) [M+H]+ requires m/z 450.03, found m/z 450.10.
|0372] To a stirred solution of 5-chloro-2-hydroxy-3-(pyridin-3-yl)-A-(6- (trifluoromethyl)benzo[t/]thiazol-2-yl)benzamide (12 mg, 0.0267 mmol) in THF (5 mL) was added 2. ON HC1 in ether (15 pL, 0.0267 mmol). The mixture was stirred at rt for 20mins. The resulting precipitate was fdtered and washed with diethyl ether to afford 5-chloro-2-hydroxy- 3-(pyridin-3-yl)-M-(6-(trifluoromethyl)benzo[d]thiazol-2-yl)benzamide hydrochloride as a yellow solid (13 mg, 100%). MS (ESI) [M+H]+ requires m/z 450.03, found m/z 450.10.
Example 11
5-chloro-3-((c/s-2,6-dimethylmorpholino)methyl)-2-hydroxy-/V-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide(l 1) and 5-chloro-3-((c7.s-2,6- dimethylmorpholino)methyl)-2-hydroxy-JV-(6-(trifluoromethyl)benzo[t/]thiazol-2- yl)benzamide hydrochloride (11 A)
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SUBSTITUTE SHEET ( RULE 26 )
[03731 Methyl 3-(dibromomethyl)-5-chloro-2-methoxybenzoate (370 mg, 1 mmol, Example 41) was dissolved in 4 mL concentrated sulfuric acid and the mixture was stirred at rt for 2h. The reaction mixture was poured into ice water and extracted with ethyl acetate two times. The combined organic layer was washed with brine and dried over sodium sulfate. After concentration, the residue was purified via silica gel column chromatography to give methyl 5-chloro-3-formyl-2-methoxybenzoate (220 mg, 96%) as a white solid. 'H NMR (300 MHz, cdch) 5 10.33 (s, 1H), 8.00 (d, J = 2.6 Hz, 1H), 7.91 (d, J = 3.1 Hz, 1H), 3.98 (s, 4H), 3.94 (s, 4H).
[0374] To a stirred solution of methyl 5-chl oro-3 -formyl-2-methoxybenzoate (220 mg, 0.965 mmol) in MeOH (10 mL) was added (25,6A)-2,6-dimethylmorpholine (238 pL, 1.93 mmol), NaBTLCN (121 mg, 1.93 mmol) and acetic acid (138 pL, 2.41 mmol). The resulting mixture was stirred at rt overnight. Saturated NaHCOs was added and extracted with DCM two times. The combined organic layer was concentrated in vacuo and the residue was purified via silica gel column chromatography to yield methyl 5-chloro-3-((cA-2,6- dimethylmorpholino)methyl)-2-methoxybenzoate as a yellow oil (232 mg, 75% yield). MS (ESI) [M+H]+ requires m/z 328.20, found m/z 328.40.
SUBSTITUTE SHEET ( RULE 26 ) [03751 To a stirred solution of methyl 5-chloro-3-((c7.s-2,6- dimethylmorpholino)methyl)-2 -methoxybenzoate (232 mg, 0.71 mmol) in MeOH (4 mL) was added 3.5 mL IN KOH solution. The resulting mixture was stirred at rt overnight. The solvent was evaporated out and 4N HC1 in dioxane (2 mL) was added to the residue. The mixture was stirred for another 10 minutes before it was concentrated and dried under vacumn. To this residue was added HBTU (322 mg, 0.852 mmol), DMF (5 mL) and DIPEA (618 pL, 3.55 mmol). The mixture was stirred for 10 minutes and then 6- (trifluoromethyl)benzo[d]thiazol-2-amine (154 mg, 0.71 mmol) was added. The resulting reaction was heated at 120 °C for 24 h. After cooling to rt, the mixture was separated between ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and concentrated in vacuo. Purification by column chromatography gave 5-chloro-3-((cA- 2,6-dimethylmorpholino)methyl)-2-hydroxy-/V-(6-(trifluoromethyl)benzo[t/]thiazol-2- yl)benzamide as a white solid (77 mg, 35%). 'H NMR (300 MHz, acetone) 8 8.41 (s, 1H), 8.00 - 7.90 (m, 2H), 7.76 (dd, J= 8.5, 2.4 Hz, 1H), 7.36 - 7.29 (m, 1H), 4.27 (s, 2H), 4.02 - 3.93 (m, 2H), 3.39-3.33 (m, 4H), 2.51 (t, J= 11.4 Hz, 2H), 1.23 (d, J= 6.3 Hz, 6H). MS (ESI) [M+H]+ requires m/z 500.10, found m/z 500.30.
|0376] To a stirred solution of 5-chloro-3-((cA-2,6-dimethylmorpholino)methyl)- 2-hydroxy-A-(6-(trifluoromethyl)benzo[t/]thiazol-2-yl)benzamide (77 mg, 0.154 mmol) in THF (5 mL) was added 2. ON HC1 in ether (77 pL, 0.154 mmol). The mixture was stirred at rt for 20 minutes. The resulting precipitate was filtered and washed with diethyl ether to afford - chi oro-3 -((cis-2, 6-dimethylmorpholino)m ethyl)-2-hy droxy-.V-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide hydrochloride as a yellow solid (82 mg, 100%). MS (ESI) [M+H]+ requires m/z 500.10, found m/z 500.30.
Example 12
5-chloro-2-hydroxy-3-(thiazol-2-yl)-A-(6-(trifluoromethyl)benzo[<7]thiazol-2- yl)benzamide(12) and 5-chloro-2-hydroxy-3-(thiazol-2-yl)-A-(6- (trifluoromethyl)benzo[ ]thiazol-2-yl)benzamide hydrochloride (12A)
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SUBSTITUTE SHEET ( RULE 26 )
[0377] A mixture of methyl 3-bromo-5-chloro-2-methoxybenzoate (277 mg, 0.99 mmol, Example 2), 2-tributylstannylthiazole (346 pL, 1.1 mmol) and Pd(PPh3)4 (35 mg, 0.03 mmol) in dioxane (2 m ) was heated to 150 °C for 20 minutes in a microwave under nitrogen. After cooling to rt, the mixture was filtered through celite, washed with ethyl acetate and concentrated in vacuo. The resulting solid was purified by flash column to obtain methyl 5 -chi oro-2-m ethoxy-3 -(thiazol -2 -yl)benzoate (237 mg, 85%) as a yellow solid. MS (ESI) [M+H]+ requires m/z 284.01, found m/z 284.0.
[0378] To a stirred solution of methyl 5-chloro-2-methoxy-3-(thiazol-2- yl)benzoate (237 mg, 0.837 mmol) in MeOH (5 mL) and THF (4 mL) was added 4.0 mb IN KOH solution. The resulting mixture was stirred at rt overnight. The solvent was evaporated out and 10% citric acid was added to the residue to pH=3. The mixture was stirred for another 10 minutes before it was concentrated and dried under vacumn. To this residue was added HBTU (379 mg, 1.0 mmol), DMF (5 mL) and DIPEA (729 pL, 4.185 mmol). The mixture was stirred for 10 minutess and then 6-(trifluoromethyl)benzo[tZ]thiazol-2-amine (182 mg, 0.837 mmol) was added. The resulting reaction was heated at 120 °C for 24h. After cooling to rt, the mixture was separated between ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and concentrated in vacuo. Purification by column
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SUBSTITUTE SHEET ( RULE 26 ) chromatography gave 5-chloro-2-hydroxy-3-(thiazol-2-yl)-/V-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide as a yellow solid (177 mg, 47%). 'H NMR (300 MHz, cdch) 5 8.28 (d, J= 2.6 Hz, 1H), 8.07 (s, 1H), 7.88 - 7.74 (m, 3H), 7.62 (d, J= 8.5 Hz, 1H), 7.42 (d, J= 3.4 Hz, 1H). MS (ESI) [M+H]+ requires m/z 455.99, found m/z 456.2.
103791 To a stirred solution of 5-chloro-2-hydroxy-3-(thiazol-2-yl)-/V-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (177 mg, 0.389 mmol) in THF (5 mL) was added 2N HC1 in ether (195 pL, 0.389 mmol). The mixture was stirred at rt for 20 minutess. The resulting precipitate was filtered and washed with diethyl ether to afford 5-chloro-2- hydroxy-3-(thiazol-2-yl)-iV-(6-(trifluoromethyl)benzo[t7]thiazol-2-yl)benzamide hydrochloride as a yellow solid (191 mg, 100%). MS (ESI) [M+H]+ requires m/z 455.99, found m/z 456.2.
Example 13
5-chloro-2-hydroxy-iV-(4-((2-methoxy ethoxy )methyl)-6-('tri fluoromethyl )benzo[c/]thiazol-2- yl)-3 -methylbenzamide (13)
SUBSTITUTE SHEET ( RULE 26 )
[0380 | A mixture of 2-methyl-4-(trifluoromethyl)aniline (525 mg, 3 mmol), NH4SCN (228 mg, 3 mmol) and TFA (574 pL, 7.5 mmol) in ethyl acetate (5 mL) was refluxed overnight. After cooling to rt, the mixture was partitioned between water and ethyl acetate. The ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give l-(2-m ethyl - 4-(trifluoromethyl)phenyl)thiourea (347 mg, 50%) as a yellow solid. MS (ESI) [M+H]+ requires m/z 235.05, found m/z 235.30.
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SUBSTITUTE SHEET ( RULE 26 ) [03811 At 0 °C, a solution of Bn (76 pL, 1.48 mmol)in CHCh (2 mL) was added dropwise to a stirred solution of l-(2-methyl-4-(trifluoromethyl)phenyl)thiourea (347 mg, 1.48 mmol) in CHCh (5 mL). After completion of the addition, the mixture was heated to reflux overnight. After cooling to rt, the precipitated out white solid was filtered and collected, neutralized by addition ammoniaand further extracted with ethyl acetate. The organic layer was dried over sodium sulfate and concentrated under reduced pressure to afford 4-methyl-6-(trifluoromethyl)benzo[d]thiazol-2-amine (125 mg, 37%) as a white solid which was used in the next step without further purification. MS (ESI) [M+H]+ requires m/z 233.04, found m/z 233.20.
[0382[ To a stirred solution of 4-rnethyl-6-(trifluoromethyl)benzo[<7]thiazol-2- amine (97 mg, 0.418 mmol) and (Boc O (262 mg, 1.2 mmol) in anhydrous DCM (4 mL) was added DMAP (5 mg, 0.0418 mmol). The resulting mixture was stirred at rt for 5h. After completion of the reaction, ethyl acetate was added and washed sequentially with saturated NH4CI, water, saturated sodium bicarbonate and brine. The organic layer was dried over sodium sulfate and concentrated under reduced pressure to afford the tert-butyl N-tert- butoxycarbonyl-JV- (4-(m ethyl )-6-(trifluoromethyl)benzo/r//thiazol-2-yl)carbamate (180 mg, 100%) as a yellow solid which was used in the next step without further purification. MS (ESI) [M+H]+ requires m/z 433.14, found m/z 433.4.
[0383| A mixture of tert-butyl A-tert-butoxycarbonyl-A- (4-(methyl)-6- (trifluoromethyl)benzo[d]thiazol-2-yl)carbamate (180 mg, 0.417 mmol), NBS (82 mg, 0.459 mmol) and AIBN (7 mg, 0.0417 mmol) in CCI4 (5 mL) was refluxed for 2.5h under nitrogen. After cooling to rt, the mixture was concentrated and the residue was subjected to chromatography to afford tert-butyl A-(4-(bromomethyl)-6-(trifluoromethyl)benzo[ ]thiazol- 2-yl)-A-tert-butoxycarbonylcarbamate (210 mg, 98%) as a yellow solid. MS (ESI) [M+H]+ requires m/z 511.05, found m/z 511.20.
|0384] To a stirred solution of tert-butyl A-(4-(bromomethyl)-6- (trifluoromethyl)benzo[t/]thiazol-2-yl)-A-tert-butoxycarbonylcarbamate (210 mg, 0.411 mmol) in 2-methoxyethan-l-ol (8 mL) was added IN NaOH (4 mL). The resulting mixture was stirred at rt for 30 minutes. After completion of the reaction, the mixture was partitioned between ethyl acetate and water. The ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give te/7-butyl (4-((2-methoxyethoxy)methyl)-6- (trifluoromethyl)benzo[t/]thiazol-2-yl)carbamate (86 mg, 52%) as a colorless oil. MS (ESI) [M+H]+ requires m/z 407.13, found m/z 407.3.
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SUBSTITUTE SHEET ( RULE 26 ) [0385| To a stirred solution of Zc/7-butyl (4-((2-methoxyethoxy)methyl)-6- (trifluoromethyl)benzo[<i]thiazol-2-yl)carbamate (86 mg, 0.212 mmol) in DCM (4 mL) was added TFA (3 mL), the resulting mixture was stirred at rt for 2h After completion of the reaction, the excess of TFA was evaporated out and the residue was partitioned between ethyl acetate and saturated sodium bicarbonate. The ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure to afford 4-((2-methoxyethoxy)methyl)-6- (trifluoromethyl)benzo[d]thiazol-2-amine as a pale yellow solid (62 mg, 97%) which was used in the subsequent step without further purification. MS (ESI) [M+H]+ requires m/z 307.07, found m/z 307.20.
[0386| A mixture of methyl 3-bromo-5-chloro-2 -methoxybenzoate (756 mg, 2.7 mmol, Example 2), methylboronic acid (324 mg, 5.4 mmol), Pd(OAc)2 (24 mg, 0.11 mmol), tricyclohexylphosphine (68 mg, 0.24 mmol) and potassium phosphate tribasic (1.9g, 8.96 mmol) was refluxed in toluene(10 mL) and water (1 mL) under N2 overnight. After the reaction was cooled, saturated NH4CI solution was added and extracted with ethyl acetate two times. The combined ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give methyl 5-chloro-2-methoxy-3-methylbenzoate (520 mg, 91%) as a yellow oil. XH NMR (300 MHz, chloroform) 5 7.62 (d, 1H, J=3.0Hz), 7.32(d, 1H, J=3.0Hz), 3.92(s, 3H), 3.82 (s, 3H), 2.30 (s, 3H).
[03871 To a stirred solution of methyl 5-chloro-2-methoxy-3-methylbenzoate (43 mg, 0.20 mmol) in MeOH (5 mL) was added 1.0 mL IN KOH solution. The resulting mixture was stirred at rt overnight. After completion of the reaction, IN HC1 was added to the reaction and extracted with ethyl acetate two times. The combined organic phase was dried over sodium sulfate and concentrated under reduced pressureto afford 5-chloro-2-methoxy-3- methylbenzoic acid as a residue (white solid, 40 mg, 100%) which was used in the next step without further purification.
[0388J 5-chloro-2-methoxy-3-methylbenzoic acid (40 mg, 0.20 mmol) was dissolved in DCM (3.0 mL), followed by the addition of acatalytic amount of DMF (1 drop) and oxalyl chloride (21 pL, 0.243 mmol) respectively. The reaction was allowed to stir at rt for 30 minutes and concentrated in vacuo. The residue was re-dissolved in THF (5.0 mL), and Hunig’s base (43 pL, 0.243 mmol) and 4-((2-methoxyethoxy)methyl)-6- (trifluoromethyl)benzo[d]thiazol-2-amine (62 mg, 0.20 mmol) were added. The mixture was stirred at rt for 48 hours before silica gel was added to quench the reaction. Solvent was evaporated and the resulting residue was purified via silica gel column chromatography to
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SUBSTITUTE SHEET ( RULE 26 ) yield 5-chloro-2-methoxy-JV-(4-((2-methoxy ethoxy )methyl)-6- (trifluoromethyl)benzo[J]thiazol-2-yl)-3-methylbenzamide (20 mg, 21%) as a white solid. MS (ESI) [M+H]+ requires m/z 489 08, found m/z 489 20.
|0389] A solution of 5-chloro-2-methoxy-JV-(4-((2-methoxy ethoxy )methyl)-6-
(trifluoromethyl)benzo[ ]thiazol -2 -yl)-3 -methylbenzamide (20 mg, 0.04 mmol) in DMF (5 mL) is mixed with sodium ethanethiolate (17 mg, 0.205 mmol) and the resulting suspension is heated at 130 °C overnight. After completion of the reaction, IN HC1 was added to the reaction and extracted with ethyl acetate two times. The combined ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give the 5-chloro-2-hydroxy-/V-(4-((2- methoxy ethoxy )methyl)-6-(trifluoromethyl)benzo[<7]thiazol-2-yl)-3 -methylbenzamide (15 mg, 55%) as a white solid. XHNMR (300 MHz, cdcl3) 57.95 (s, 2H), 7.58 (s, 1H), 7.29 (s, 1H), 4.96 (s, 2H), 3.98 - 3.86 (m, 2H), 3.84 - 3.73 (m, 2H), 3.64 (s, 3H), 2.28 (s, 3H). MS (ESI) [M+H]+ requires m/z 475.06, found m/z 475.20.
Example 14
5-chl oro-2 -hydroxy-3-(tetrahydrofuran-3-yl)-/V-(6-(trifluorornethyl)benzo[d]thi azol -2- yl)benzamide (14)
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SUBSTITUTE SHEET ( RULE 26 )
[0390| A mixture of methyl 3-bromo-5-chloro-2-methoxybenzoate (275 mg, 0.98 mmol, Example 2), furan-3-ylboronic acid (218 mg, 1.96 mmol), Pd(PPhi)4 (56 mg, 0.049 mmol) and sodium carbonate (312 mg, 2.95 mmol) in dioxane (3 mL) and water (1 mL) was irradiated under microwave at 100 °C for Ih under nitrogen. After cooling to it, the mixture was partitioned between water and ethyl acetate. The ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give methyl 5-chl oro-3 -(furan-3-yl)-2 -methoxybenzoate (250 mg, 96%) as a colorless oil. MS (ESI) [M+H]+ requires m/z 267.03, found m/z 267.20.
(0391] A mixture of methyl 5-chloro-3-(furan-3-yl)-2-methoxybenzoate (267 mg, 1.0 mmol) and Pd-C (27 mg) in MeOH (5 mL) was stirred at rt under hydrogen atmosphere for 5h. The reaction mixture was filtered and the filtrate was concentrated to afford methyl 2- methoxy-3-(tetrahydrofuran-3-yl)benzoate (238 mg, 100%) as a colorless oil, which was used directly in the next step without further purification. MS (ESI) [M+H]+ requires m/z 237.1, found m/z 237.3.
[0392] A mixture of methyl 2-methoxy-3-(tetrahydrofuran-3-yl)benzoate (238 mg, 1 mmol) and NCS (134 mg, 1.0 mmol) in acetonitrile (5 mL) was refluxed for 24h. After cooling to rt, the solvent was concentrated and the residue was subjected to column directly to afford methyl 5-chloro-2-methoxy-3-(tetrahydrofuran-3-yl)benzoate (85 mg, 31%) as a colorless oil. 'HNMR (300 MHz, cdch) 8 7.66 - 7.60 (m, IH), 7.42 (d, J= 2.7 Hz, IH), 4.11
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SUBSTITUTE SHEET ( RULE 26 ) - 4.03 (m, 2H), 3.90 (s, 3H), 3.86 - 3.74 (m, 5H), 3.72 - 3.64 (m, 1H), 2.44 - 2.29 (m, 1H), 1.98 - 1.84 (m, 1H). MS (ESI) [M+H]+ requires m/z 271.07, found m/z 271.2.
|0393] To a stirred solution of 5-chloro-2-methoxy-3-(tetrahydrofuran-3- yl)benzoate (85 mg, 0.315 mmol) in MeOH (5 mL) was added 2.0 mL IN KOH solution. The resulting mixture was stirred at rt overnight. The solvent was evaporated out and 4N HC1 in dioxane (1 mL) was added to the residue The mixture was stirred for another 10 minutes before it was concentrated and dried under vacumn. To this residue was added HBTU (143 mg, 0.378 mmol), DMF (5 mL) and DIPEA (274 pL. 1.575 mmol). The mixture was stirred for 10 minutes and then 6-(trifluoromethyl)benzo[<7]thiazol-2-amine (68 mg, 0.315 mmol) was added. The resulting reaction was heated at 120 °C for 24h. After cooling to rt, the mixture was separated between ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and concentrated in vacuo. Purification by column chromatography gave 5 -chloro-2-hy droxy-3 -(tetrahy drofuran-3 -y 1 )- N- (6 - (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide as a white solid (14 mg, 10%). 'H NMR (400 MHz, acetone) 6 8.42 (s, 1H), 8.12 (d, J= 2.6 Hz, 1H), 7.91 (d, J= 8.5 Hz, 1H), 7.83 (dd, J= 8.5, 1.4 Hz, 1H), 7.47 (d, J= 2.6 Hz, 1H), 4.09 - 3.96 (m, 2H), 3.88 - 3.71 (m, 3H), 2.37 - 2.31 (m, 1H), 1.98 - 1.84 (m, 1H). MS (ESI) [M+H]+ requires m/z 443.04, found m/z 443.3.
Example 15
5-chl oro-2 -hy droxy-3-(tetrahydrofuran-2-yl)-/V-(6-(trifluoromethyl)benzo[ ]thi azol -2- yl)benzamide (15)
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SUBSTITUTE SHEET ( RULE 26 )
[0394] A mixture of methyl 3-bromo-5-chloro-2-methoxybenzoate (253 mg, 0.90 mmol, Example 2), furan-2-ylboronic acid (132 mg, 1.17 mmol), Pd(PPh3)4 (52 mg, 0.045 mmol) and sodium carbonate (287 mg, 2.7 mmol) in dioxane (3 mL) and water (1 mL) was irradiated under microwave at 100 °C for Ih under nitrogen. After cooling to rt, the mixture was partitioned between water and ethyl acetate. The ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give methyl 5-chl oro-3 -(furan -2 -yl)-2 -methoxybenzoate (131 mg, 55%) as a colorless oil. MS (ESI) [M+H]+ requires m/z 267.03, found m/z 267.20.
[0395] A mixture of methyl 5-chl oro-3 -(furan-2-yl)-2-methoxybenzoate (131 mg, 0.5 mmol) and Pd-C (15 mg) in MeOH (5 mL) was stirred at rt under hydrogen atmosphere for 6h. The reaction mixture was filtered and the filtrate was concentrated, the residue was purified via silica gel column chromatography to give methyl 5-chloro-2-methoxy-3- (tetrahydrofuran-2-yl)benzoate (95 mg, 70%) as a colorless oil. 'H NMR. (300 MHz, cdch) 8 7.71 (d, 7= 2.8 Hz, IH), 7.62 (dd, 7= 2.8, 0.6 Hz, IH), 5.13 (t, 7= 7.2 Hz, IH), 4.16 - 4.08 (m, IH), 3.99 - 3.90 (m, IH), 3.93 (s, 3H), 3.84 (s, 3H), 2.52 - 2.38 (m, IH), 2.07 - 1.96 (m, 2H), 1.73 - 1.65 (m, IH). MS (ESI) [M+H]+ requires m/z 271.07, found m/z 271.2.
[0396| To a stirred solution of 5-chloro-2-methoxy-3-(tetrahydrofuran-2- yl)benzoate (95 mg, 0.352 mmol) in MeOH (5 mL) was added 2.0 mL IN KOH solution. The resulting mixture was stirred at rt overnight. The solvent was evaporated out and 4N HC1 in dioxane (1 mL) was added to the residue. The mixture was stirred for another 10 minutes
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SUBSTITUTE SHEET ( RULE 26 ) before it was concentrated and dried under vacumn. To this residue was added HBTU (160 mg, 0.422 mmol), DMF (5 mL) and DIPEA (306 pL, 1.76 mmol). The mixture was stirred for 10 minutes and then 6-(trifluoromethyl)benzo[<7]thiazol-2-amine (76 mg, 0.352 mmol) was added. The resulting reaction was heated at 120 °C for 24h. After cooling to rt, the mixture was separated between ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and concentrated in vacuo. Purification by column chromatography gave 5-chl oro-2-hydroxy-3-(tetrahydrofuran-2-yl)-A-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide as a white solid (15 mg, 10%). 'H NMR (500 MHz, acetone) 5 8 42 (s, 1H), 8.11 (s, 1H), 7.91 (d, J= 8.3 Hz, 1H), 7.82 (d, J= 8.5 Hz, 1H), 7.56 (s, 1H), 5.12 (t, J= 7.1 Hz, 1H), 4.17 - 4.10 (m, 1H), 3.94 - 3.85 (m, 1H), 2.55 - 2.45 (m, 1H), 2.03 - 1.91 (m, 2H), 1.78 - 1.66 (m, 1H). MS (ESI) [M+H]+ requires m/z 443.04, found m/z 443.3.
Example 16
5-chloro-2-hydroxy-3-(morpholinomethyl)-jV-(6-(trifluoromethyl)benzo[6?]thiazol-2- yl)benzamide(16) and 5-chl oro-2 -hy droxy-3-(morpholinomethyl)-A'-(6- (trifluoromethyl)benzo[6?]thiazol-2-yl)benzarnide hydrochloride (16A)
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SUBSTITUTE SHEET ( RULE 26 )
[0397| To a flame dried flask was added NBS (183 mg, 1.03 mmol), AIBN (15 mg, 0.093 mmol) and a solution of methyl 5-chloro-2-methoxy-3-methylbenzoate (201g, 0.93 mmol) in CCh (10 mL). The suspension was refluxed in the dark overnight. The mixture was cooled to rt and concentrated. The residue was purified via silica gel column chromatography to give methyl 5-chloro-3-(bromomethyl)-2-methoxybenzoate as a colorless oil (236 mg, 87%). 'l l NMR (300 MHz, cdch) 57.76 (d, J = 2.7 Hz, 1H), 7.54 (d, J = 2.7 Hz, 1H), 4.51 (s, 2H), 3.96 (s, 3H), 3.94 (s, 3H).
[0398] At 0 °C, to a stirred solution of methyl 5-chl oro-3 -(bromomethyl)-2- methoxybenzoate (132 mg, 0.45 mmol) in THF (4 mL) was added morpholine (78 pL, 0.90 mmol) and the mixture was stirred at rt for 6h. After completion of the reaction, the reaction was partitioned between DCM and water. The organic layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by flash column to afford methyl 5 -chi oro-2-m ethoxy-3 -(morpholinomethyl)benzoate (110 mg, 83%) as a colorless oil.
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SUBSTITUTE SHEET ( RULE 26 ) XHNMR (500 MHz, cdcl3) 8 7.68 (d, J= 2.8 Hz, 1H), 7.58 (d, J= 2.7 Hz, 1H), 3.90 (s, 3H), 3.82 (s, 3H), 3.70 (t, 4H), 3.52 (s, 2H), 2.47 (t, J= 4.1 Hz, 4H). MS (ESI) [M+H]+ requires m/z 300.10, found m/z 300.30.
|0399] To a stirred solution of methyl 5-chloro-2-methoxy-3- (morpholinomethyl)benzoate (110 mg, 0.368 mmol) in MeOH (3 mL) was added 2.0 mL IN KOH solution. The resulting mixture was stirred at rt overnight. The solvent was evaporated out and 4N HC1 in dioxane (1 mL) was added to the residue. The mixture was stirred for another 10 minutes before it was concentrated and dried under vacumn. To this residue was added HBTU (168 mg, 0.442 mmol), DMF (5 mL) and DIPEA (320 pL, 1.84 mmol) The mixture was stirred for 10 minutes and then 6-(trifluoromethyl)benzo[tZ]thiazol-2-amine (80 mg, 0.368 mmol) was added. The resulting reaction mixturewas heated at 120 °C for 24h. After cooling to rt, the mixture was separated between ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and concentrated in vacuo. Purification by column chromatography gave 5-chloro-2-hydroxy-3-(morpholinomethyl)-7V- (6-(trifluoromethyl)benzo[t/]thiazol-2-yl)benzamide (75 mg, 43%) as a yellow solid. ’H NMR (400 MHz, dmso) 8 8.39 (s, 1H), 7.80 (dd, J= 9.5, 5.7 Hz, 2H), 7.67 (d, J= 8.6 Hz, 1H), 7.37 (d, J= 2.9 Hz, 1H), 4.23 (s, 2H), 3.81 (brs, 4H), 3.17 (brs, 4H). MS (ESI) [M+H]+ requires m/z 472.07, found m/z 472.20.
[04(H)] To a stirred solution of 5-chloro-2-hydroxy-3-(morpholinomethyl)-A-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (32 mg, 0.068 mmol) in THF (5 mL) was added 4. ON HC1 in dioxane (20 pL, 0.075 mmol). The mixture was stirred at rt for 20 minutes. The resulting precipitate was filtered and washed with diethyl ether to afford 5- chloro-2-hydroxy-3-(morpholinomethyl)-7V-(6-(trifluoromethyl)benzo[<7]thiazol-2- yl)benzamide hydrochloride as a yellow solid (100%). MS (ESI) [M+H]+ requires m/z 472.07, found m/z 472.20.
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SUBSTITUTE SHEET ( RULE 26 ) Example 17
5-chloro-2-hydroxy-3-(2-morpholinoethyl)-A-(6-(trifluoromethyl)benzo[</]thiazol-2- yl)benzamide(17) and 5-chl oro-2 -hy droxy-3-(2-morpholinoethyl)-A-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide hydrochloride (17A)
[0401] A mixture of methyl 3-bromo-5-chloro-2-methoxybenzoate (445 mg, 1.6 mmol, Example 2), potassium trifluoro(vinyl)borate (322 mg, 2.4 mmol), Pd(dppf)C12, (40
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SUBSTITUTE SHEET ( RULE 26 ) mg, 0.05 mmol), and sodium carbonate (339 mg, 3.2 mmol) was refluxed in dioxane (10 mL) and water (1 mL) under N2 overnight. After the reaction was cooled, saturated NH4CI solution was added and extracted with ethyl acetate two times. The combined ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give methyl 5-chloro-2-methoxy-3- vinylbenzoate (320 mg, 89%) as a colorless oil. XHNMR (300 MHz, cdch) 57.69 (d, J= 2.7 Hz, 1H), 7.63 (d, .7= 2,7 Hz, 1H), 6.99 (dd, J= 17.7, 11.1 Hz, 1H), 5.79 (dd, J= 17.7, 0.8 Hz, 1H), 5.43 (dd, J= 11.1, 0.8 Hz, 1H), 3.93 (s, 3H), 3.83 (s, 3H).
[0402] At 0 °C, to a stirred solution of methyl 5-chloro-2-methoxy-3- vinylbenzoate (270 mg, 1.19 mmol) was added 9-BBN (3.1 mL, 1.55 mmol, 0.5M in THF) dropwise. After completion of the addition, the mixture was allowed to stir at rt overnight. The reaction was cooled to 0 °C and 1 mL of 50% H2O2 solution was added followed by 1 mL of 3M NaOH solution. The mixture was stirred over an ice bath for 2h. The reaction was partitioned between water and ethyl acetate. The ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give methyl 5-chloro-3-(2-hydroxyethyl)-2-methoxybenzoate (110 mg, 50%) as a colorless oil. 'H NMR (400 MHz, d2o) 5 7.65 (s, 1H), 7.37 (s, 1H), 3.90 (s, 3H), 3.85 - 3.78 (m, 5H), 2.88 (t, J= 6.3 Hz, 2H). MS (ESI) [M+H]+ requires m/z 245.06, found m/z 245.20.
[0403] At 0 °C, to a stirred solution of methyl 5-chl oro-3 -(2 -hydroxy ethyl)-2- methoxybenzoate (110 mg, 0.451 mmol) in DCM (3 mL) was added TEA (95 pL, 0.677 mmol) and MsCl (42 pL, 0.54 mmol). The resulting mixture was stirred at 0 °C for Ih. The reaction mixture was partitioned between DCM and saturated NH4CI solution. The organic layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give methyl 5-chloro-2-methoxy-3-(2- ((methylsulfonyl)oxy)ethyl)benzoate (119 mg, 83%) as a colorless oil. 'H NMR (500 MHz, cdch) 8 7.71 (d, J= 2.7 Hz, IH), 7.37 (d, J= 2.7 Hz, IH), 4.41 (t, J= 6.8 Hz, 2H), 3.92 (s, 3H), 3.83 (s, 3H), 3.08 (t, J= 6.8 Hz, 2H), 2.93 (s, 3H).
[0404] To a stirred solution of methyl 5-chloro-2-methoxy-3-(2- ((methylsulfonyl)oxy)ethyl)benzoate (119 mg, 0.369 mmol) in DMF (3 mL) was added morpholine (160 pL, 1.84 mmol). The resulting mixture was stirred at 80 °C overnight. The reaction was partitioned between water and DCM. The organic layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give methyl 5-chloro-2-methoxy-3-(2-morpholinoethyl)benzoate
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SUBSTITUTE SHEET ( RULE 26 ) (91 mg, 83%) as a yellow oil. 'H NMR (300 MHz, cdch) 57.65 (d, J= 2.3 Hz, 1H), 7.35 (d, 1H), 3.91 (s, 3H), 3.82 (s, 3H), 3.78 - 3.69 (m, 4H), 2.86 - 2.79 (m, 2H), 2.64 - 2.56 (m, 2H), 2.57 - 2.49 (m, 4H) MS (ESI) [M+H]+ requires m/z 314 12, found m/z 314 20.
|0405] To a stirred solution of methyl 5-chloro-2-methoxy-3-(2- morpholinoethyl)benzoate (91 mg, 0.29 mmol) in MeOH (3 mL) was added 1.5 mL IN KOH solution. The resulting mixture was stirred at rt overnight. The solvent was evaporated out and 4N HC1 in dioxane (1 mL) was added to the residue. The mixture was stirred for another 10 minutes before it was concentrated and dried under vacumn. To this residue was added HBTU (132 mg, 0.35 mmol), DMF (5 mL) and DIPEA (253 pL, 1. 45 mmol). The mixture was stirred for 10 minutes and then 6-(trifluoromethyl)benzo[t/]thiazol-2-amine (64 mg, 0.29 mmol) was added. The resulting reaction was heated at 120 °C for 24h. After cooling to rt, the mixture was separated between ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and concentrated in vacuo. Purification by column chromatography gave 5-chloro-2-hydroxy-3-(2-morpholinoethyl)-/V-(6- (trifluoromethyl)benzo[< ]thiazol-2-yl)benzamide (56 mg, 40%) as a yellow solid. H NMR (300 MHz, acetone) 6 8.41 - 8.36 (m, 1H), 7.94 - 7.86 (m, 2H), 7.77 - 7.70 (m, 1H), 7.24 (d, J= 2.9 Hz, 1H), 4.04 (brs, 4H), 3.40 - 3.02 (m, 8H). MS (ESI) [M+H]+ requires m/z 486.10, found m/z 486.20.
[0406] To a stirred solution of 5-chloro-2-hydroxy-3-(2-morpholinoethyl)-/V-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (36 mg, 0.074 mmol) in THF (5 mL) was added 4. ON HC1 in dioxane (21 pL, 0.082 mmol). The mixture was stirred at rt for 20 minutes. The resulting precipitate was filtered and washed with diethyl ether to afford 5- chloro-2-hydroxy-3-(2-morpholinoethyl)-A-(6-(trifluoromethyl)benzo[c ]thiazol-2- yl)benzamide hydrochloride as a yellow solid (100%). MS (ESI) [M+H]+ requires m/z 486.10, found m/z 486.20.
Example 18
5-chloro-/V-(2-chloro-4-(trifluoromethyl)phenyl)-3-(2-(dimethylamino)ethyl)-2- hydroxybenzamide (18)
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SUBSTITUTE SHEET ( RULE 26 )
[0407| Methyl 3-bromo-5-chloro-2-methoxybenzoate (6.18g, 22.07 mmol, Example 2), potassium (2-((fer/-butoxycarbonyl)amino)ethyl)trifluoroborate (16.6g, 66.21 mmol), Pd(dppf)C12 (968 mg, 1.32 mmol) and CS2CO3 (21.5g, 66.21 mmol) were added to a flask. This flask was evacuated and refilled with N2 three times. Subsequently, toluene (75 mL) and water (25 mL) were added to the flask under N2. The mixture was stirred at 80 °C under N2 overnight. Saturated NH4CI solution was added and the resulting mixture was extracted with ethyl acetate two times. The combined organic layer was concentrated in vacuo and the residue was purified via silica gel column chromatography to yield methyl 3- (2-((ter/-butoxycarbonyl)amino)ethyl)-5-chloro-2-methoxybenzoate as a yellow oil (1.51g, 20% yield). 1H NMR (300 MHz, cdcl3) 5 7.64 (d, J = 2.7 Hz, 1H), 7.31 (d, J = 2.7 Hz, 1H), 3.90 (s, 3H), 3.81 (s, 3H), 3.34 (brs, 2H), 2.82 (t, J = 6.9 Hz, 2H), 1.41 (s, 9H). MS (ESI) [M+Na]+requires m/z 366.11, found m/z 365.95.
[0408] Methyl 3 -(2-((tert-butoxy carbonyl )ami no)ethyl )-5-chl oro-2 - methoxybenzoate (1.12g, 3.26 mmol) was dissolved in 4N HC1 in dioxane (5 mL) and the
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SUBSTITUTE SHEET ( RULE 26 ) resulting mixture was stirred at rt for 2h. Saturated NaHCCh solution was added and extracted with DCM. The organic layer was washed with water and brine and dried over sodium sulfate. The organic layer was filtered and the solvent removed in vacuo to yield methyl 3-(2-aminoethyl)-5-chloro-2-methoxybenzoate as a pale yellow oil (475 mg, 60%). 1H NMR (500 MHz, cdcl3) 8 7.61 (d, J = 2.7 Hz, 1H), 7.30 (d, J = 2.7 Hz, 1H), 3.87 (s, 3H), 3.78 (s, 3H), 2.92 (t, J = 7.5 Hz, 2H), 2.75 (t, J = 7.1 Hz, 2H), 1.61 (s, 2H). ). MS (ESI) [M+H]+ requires m/z 244.08, found m/z 244.30.
|0409] To a stirred solution of methyl 3-(2-aminoethyl)-5-chloro-2- methoxybenzoate (100 mg, 0.411 mmol) in MeOH (3 mL) was added formaldehyde (122 pL, 1.64 mmol, 37% wt in H2O), NaBHsCN (103 mg, 1.64 mmol) and acetic acid (117 pL, 2.06 mmol). The resulting mixture was stirred at rt overnight. Saturated NaHCCh was added and extracted with DCM two times. The combined organic layer was concentrated in vacuo and the residue was purified via silica gel column chromatography to yield methyl 5-chloro-3-(2- (dimethylamino)ethyl)-2-methoxybenzoate as a yellow oil (88 mg, 82% yield). 'H NMR (300 MHz, cdcl3) 87.66 (d, J = 2.5 Hz, 1H), 7.36 (d, J = 2.6 Hz, 1H), 3.92 (s, 3H), 3.84 (s, 3H), 2.86 (t, J = 7.5 Hz, 2H), 2.58 (t, J = 7.5 Hz, 2H), 2.34 (s, 6H). MS (ESI) [M+H]+ requires m/z 272.10, found m/z 272.05.
[0410] To a stirred solution of methyl 5-chl oro-3 -(2-(dimethylamino)ethyl)-2- methoxybenzoate (82 mg, 0.3 mmol) in MeOH (3 mL) was added IN KOH (1 mL) solution. The mixture was stirred at rt overnight. IN HC1 was added to adjust the pH to 1. The mixture was concentrated in vacuo. The residue was dissolved in DCM (3 mL) followed by the addition of catalytic amount of DMF (1 drop) and oxalyl chloride (32 pL, 0.36 mmol) respectively. The reaction was allowed to stir at rt for 2h then concentrated in vacuo. The residue was re-dissolved in THF (5.0 mL), and Hunig’s base (157 pL, 0.90 mmol) and 2- chloro-4-(trifluoromethyl)aniline (42 pL, 0.30 mmol) were added. The mixture was stirred at rt for 48 hours before silica gel was added to quench the reaction. Solvent was evaporated and the resulting residue was purified via silica gel column chromatography to yield 5- chloro-/V-(2-chloro-4-(trifluoromethyl)phenyl)-3-(2-(dimethylamino)ethyl)-2- methoxybenzamide (124 mg, 95%) as a white solid. 'H NMR (300 MHz, cdch) 8 10.61 (s, 1H), 8.85 (d, J= 8.7 Hz, 1H), 8.02 (d, J= 2.7 Hz, 1H), 7.71 (s, 1H), 7.60 (d, J= 8.7 Hz, 1H), 7.45 (d, J= Hz, 1H), 3.92 (s, 3H), 3.04 - 2.79 (m, 2H), 2.77 - 2.49 (m, 2H), 2.42 (s, 6H). MS (ESI) [M+H]+ requires m/z 435.09, found m/z 435.0.
[0411] At -78 °C, to a stirred solution of 5-chloro-7V-(2-chloro-4- (trifluoromethyl)phenyl)-3-(2-(dimethylamino)ethyl)-2-methoxybenzamide (69 mg, 0.16
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SUBSTITUTE SHEET ( RULE 26 ) mmol) in anhydrous DCM (5 mL) was added BBr3 (1.0M in DCM, 0.635 m ) dropwise. After addition, the reaction was allowed to warm to rt slowly and the mixture was stirred at rt for 2h. After completion of the reaction, the reaction mixture was cooled in an ice bath and MeOH and water were added to quench the reaction. The mixture was separated between DCM and water. The organic layer was washed with water, brine and dried over sodium sulfate, concentrated in vacuo. The residue was purified via silica gel column chromatography to afford 5-chloro-A-(2-chloro-4-(trifluoromethyl)phenyl)-3-(2- (dimethylamino)ethyl)-2-hydroxybenzamide (56 mg, 85%) as a yellow solid. LH NMR (300 MHz, cdcl3) 5 12.50 (s, 1H), 8.96 - 8.83 (m, 1H), 8.05 (d, J= 2.8 Hz, 1H), 7.65 (d, J= 1.4 Hz, 1H), 7.55 (dd, J= 8.7, 1.5 Hz, 1H), 7.11 (d, J= 2.8 Hz, 1H), 2.91 (s, 4H), 2.62 (s, 6H). MS (ESI) [M+H]+ requires m/z 421.07, found m/z 421.20.
Example 19
3 -((4-acetylpiperazin-l-yl)methyl)-5-chl oro-2 -hydroxy -N-(6- (trifluoromethyl)benzo[t/]thiazol-2-yl)benzamide (19) and 3-((4-acetylpiperazin-l- yl)methyl)-5-chloro-2-hydroxy-A-(6-(trifluoromethyl)benzo[</]thiazol-2-yl)benzamide hydrochloride (19A)
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SUBSTITUTE SHEET ( RULE 26 )
|0412] At 0 °C, to a stirred solution of methyl 3-(bromomethyl)-5-chloro-2- methoxybenzoate (257 mg, 0.88 mmol, Example 2) in THF (5 mL) was added 1 -(piperazin- 1- yl)ethan-l-one (225 mg, 1.754 mmol) and the mixture was stirred at rt for 6h. After completion of the reaction, the reaction was partitioned between DCM and water. The organic layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by flash column to afford methyl 3-((4-acetylpiperazin-l-yl)methyl)-5- chloro-2-methoxybenzoate (265 mg, 89%) as a colorless oil. 'H NMR (300 MHz, cdch) 5 7.62 (d, J= 2.8 Hz, 1H), 7.51 (d, J = 2.8 Hz, 1H), 3.84 (s, 3H), 3.75 (s, 3H), 3.57 - 3.52 (m, 2H), 3.48 (s, 2H), 3.42 - 3.37 (m, 2H), 2.43 - 2.36 (m, 4H), 2.00 (s, 3H).
|0413] To a stirred solution of methyl 3-((4-acetylpiperazin-l-yl)methyl)-5- chloro-2-methoxybenzoate (265 mg, 0.78 mmol) in MeOH (5 mL) was added 4.0 mL IN KOH solution. The resulting mixture was stirred at rt overnight. The solvent was evaporated out and 4N HC1 in dioxane (2 mL) was added to the residue. The mixture was stirred for another 10 minutes before it was concentrated and dried under vacumn. To this residue was added HBTU (356 mg, 0.94 mmol), DMF (5 mL) and DIPEA (680 pL, 3.9 mmol). The mixture was stirred for 10 minutes and then 6-(trifluoromethyl)benzo[tZ]thiazol-2-amine (170 mg, 0.78 mmol) was added. The resulting reaction was heated at 120 °C for 24h. After cooling to rt, the mixture was separated between ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and concentrated in vacuo. Purification by column chromatography gave 3-((4-acetylpiperazin-l-yl)methyl)-5-chloro-2-hydroxy-A-(6- (trifluoromethyl)benzo[t/]thiazol-2-yl)benzamide (101 mg, 30%) as a yellow solid. 'H NMR (300 MHz, cdch) 5 8.19 (d, J= 2.6 Hz, 1H), 8.15 (s, 1H), 7.90 (d, J= 8.6 Hz, 1H), 7.70 (d, J
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SUBSTITUTE SHEET ( RULE 26 ) = 8.5 Hz, 1H), 7.26 - 7.20 (m, 1H), 3.57 - 3.52 (m, 2H), 3.48 (s, 2H), 3.42 - 3.37 (m, 2H), 2.43 - 2.36 (m, 4H), 2.00 (s, 3H). MS (ESI) [M+H]+ requires m/z 513.10, found m/z 513.30.
|0414] To a stirred solution of 3-((4-acetylpiperazin-l-yl)methyl)-5-chloro-2- hydroxy-/V-(6-(trifluoromethyl)benzo[^/]thiazol-2-yl)benzamide (101 mg, 0.197 mmol) in THF (5 mL) was added 2N HC1 in ether (100 pL, 0.2 mmol). The mixture was stirred at rt for 20 minutess. The resulting precipitate was fdtered and washed with diethyl ether to afford 3- ((4-acetylpiperazin- l-yl)methyl )-5-chloro-2-hydroxy-A',-(6-(trifluoromethyl)benzo[<7]thiazol- 2-yl)benzamide hydrochloride as a yellow solid (100%). MS (ESI) [M+H]+ requires m/z 513 10, found m/z 513 30.
Example 20
5-chloro-2-hydroxy-3-((2 -methoxy ethoxy )methyl)-JV-(6-(trifluoromethyl)pyri din-3- yl)benzamide (20) and 5-chloro-2-hydroxy-3-((2-methoxy ethoxy )methyl)-jV-(6- (trifluoromethyl)pyridin-3-yl)benzamide hydrochloride (20A)
(0415] To a solution of 5-chloro-2-methoxy-3-((2-methoxyethoxy)methyl)benzoic acid (200 mg, 728.08 pmol, Example 2), 6-(trifluoromethyl)pyridin-3-amine (118 mg, 728.08 pmol) in pyridine (2 mL) was added POCh (167 mg, 1.09 mmol, 101.49 pL) at 0 °C. The mixture was stirred at 0 °C for 1 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with water (3 mL) and extracted with DCM (3 mL x 3). The combined organic layers were washed with brine 5 mL, dried over NazSCU, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCh, petroleum ether/ethyl acetate = 3:1) to afford 5-chloro-2-
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SUBSTITUTE SHEET ( RULE 26 ) methoxy-3-((2-methoxyethoxy)methyl)-A-(6-(trifluoromethyl)pyridine-3-ylbenzamide (117 mg, 212.32 pmol, 29.16% yield, 76% purity) as a white solid. MS (ESI) [M+H]+ requires m/z 419.09, found m/z 4192
|0416] To 5-chloro-2-methoxy-3-((2-methoxyethoxy)methyl)-/V-(6- (trifluoromethyl)pyridine-3-ylbenzamide (95 mg, 226.84 pmol) in DMF (2 mL) was added NaSEt (38 mg, 453.68 pmol) and the mixture was stirred at 160 °C for 2h The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 5-chloro-2-hydroxy-3-((2- methoxyethoxy)methyl)-Ar-(6-(trifluoromethyl)pyridin-3-yl)benzamide (32.99 mg, 73 67 pmol, 32.48% yield, 98.537% purity, HC1) as a brown solid. MS (ESI) [M+H]+ requires m/z 405.08, found m/z 405.0. XHNMR (400 MHz, METHANOL-d4) 5 ppm 8.93 (d, J = 2.19 Hz, 1 H), 8.37 (dd, J = 8.33, 2.19 Hz, 1 H), 7.90 (d, J = 2.19 Hz, 1 H), 7.74 (d, J = 8.77 Hz, 1 H), 7.52 (d, J = 2.63 Hz, 1 H), 4.53 (s, 2 H), 3.60 - 3.66 (m, 2 H), 3.51 - 3.56 (m, 2 H), 3.22 (dt, J = 3.29, 1.43 Hz, 3 H).
Example 21
5-chl oro-2 -hy droxy-3-((2-methoxy ethoxy)methyl)-A-(2-(trifluoromethyl)pyrimidin-5- yl)benzamide (21)
|0417] Using a method similar to Example 20, 5-chloro-2-hydroxy-3-((2- methoxyethoxy)methyl)-/V-(2-(trifluoromethyl)pyrimidin-5-yl)benzamide was obtained. MS (ESI) [M+H]+ requires m/z 406.07, found m/z 406.0. 'HNMR (400 MHz, METHANOL-d4) 5 ppm 9.35 (s, 2 H), 8.00 (d, J = 2.43 Hz, 1 H), 7.64 (s, 1 H), 4.64 (s, 2 H) 3.73 (dd, J = 5.62, 3.42 Hz, 2 H), 3.60 - 3.66 (m, 2 H), 3.40 (s, 3 H).
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SUBSTITUTE SHEET ( RULE 26 ) Example 22
5-chloro-2-hydroxy-3-(((2-methoxyethyl)amino)methyl)-A-(6- (trifluoromethy l)b cnzo[t/] th i azol -2-y l)b enzami de (22)
|0418] A solution of 5-chloro-2-hydroxybenzoic acid (10 g, 57.95 mmol), and
6,7,8,9-tetrazatricyclodecane (16.2 g, 115.90 mmol) in TFA (100 mL) was stirred at 100 °C for 12 h, then cooled to 20 °C, followed by addition of HCI (100 mL, 3M). The reaction mixture was stirred for another 1 h, then concentrated under reduced pressure. The white residue was suspend in water (50 mL). The solid was collected and dried under vacuum to obtained 5-chloro-3-formyl-2 -hydroxybenzoic acid (6 g, crude) as a white solid. 'H NMR (400 MHz, DMSO-dg) 8 ppm 10.30 (s, 1 H), 8.02 (br d, J = 2.45 Hz, 1 H), 7.83 (br d, J = 2.93 Hz, 1 H)
|0419| A mixture of 5-chloro-3-formyl-2 -hydroxybenzoic acid (2 g, 9.97 mmol), 6-(trifluoromethyl)benzo[<7]thiazol-2-amine (2.1 g, 9.97 mmol) and EDCI (2.2 g, 11.97 mmol), HOBt (2.0 g, 14.96 mmol) in DCM (20 mL) was heated at 50 °C for 8 h. The reaction mixture was concentrated to remove solvent, then H2O (20 mL) was added and NaOH (3M) was added until the pH was- 9-10. The reaction mixture was extracted with ethyl acetate (30 mL x 2), then concentrated to remove ethyl acetate. The residue was diluted with ethyl acetate (30 mL), solid precipitated, filtered and concentrated under reduced pressure to give 5-chloro-3-formyl-2-hydroxy-A-(6-(trifluoromethyl)benzo[ri]thiazol-2- yl)benzamide (580 mg, 5.8% yield) as a yellow solid. LH NMR (400 MHz, DMSO-de) 8 ppm
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SUBSTITUTE SHEET ( RULE 26 ) 10.34 (s, 1 H), 8.43 (s, 1 H), 7.95 (d, J = 3.09 Hz, 1 H), 7.86 (d, J = 8.38 Hz, 1 H), 7.70 (br d, J = 8.60 Hz, 1 H), 7.49 (d, J = 3.31 Hz, 1 H). MS (ESI) [M+H]+ requires m/z 400.99, found m/z 401.1.
|0420] To a solution of 2-methoxyethanamine (22 mg, 299.43 pmol), 5-chloro-3- formyl-2-hydroxy-A-(6-(trifluoromethyl)benzo[J]thiazol-2-yl)benzamide (100 mg, 249.53 pmol) in MeOH (4 mb) was added acetic acid (3 mg, 49.91 pmol, 2.85 pL) to make pH ~ 5- 6. The mixture was stirred at 20 °C for 2 h, then NaBFECN (94 mg, 1.50 mmol) was added. The mixture was stirred at 20 °C for another 10 h. The reaction mixture was concentrated to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 5-chloro- 2-hydroxy-3-(((2-methoxyethyl)amino)methyl)-jV-(6-(trifluoromethyl)benzo[d]thiazol-2- yl)benzamide (28.30 mg, 57.02 pmol, 22.85% yield, 100% purity, HC1) as a brown solid. 'H NMR (400 MHz, METHANOL-d4) 8 ppm 8.27 (s, 1 H), 8.20 (d, J = 2.45 Hz, 1 H), 7.74 - 7.82 (m, 2 H), 7.63 (d, J = 2.45 Hz, 1 H), 4.31 (s, 2 H), 3.67 - 3.72 (m, 2 H), 3.44 (s, 3 H), 3.28 (d, J = 4.89 Hz, 2 H). MS (ESI) [M+H]+ requires m/z 460.06, found m/z 460.2
Example 23
5-chloro-2-hydroxy-3-(pyrrolidin-l-ylmethyl)-JV-(6-(trifluoromethyl)benzo[</]thiazol-2-yl) benzamide (23)
[0421] 5-chl oro-2 -hydroxy-3-(pyrrolidin-l-ylmethyl)-7V-(6-
(trifluoromethyl)benzo[d]thiazol-2-yl) benzamide was made in a similar manner as the described in Example 22. MS (ESI) [M+H]+ requires m/z 456.07, found m/z 456.2. 'H NMR (400 MHz, METHANOL-d4) 8 ppm 8.16 - 8.32 (m, 2 H), 7.73 - 7.83 (m, 2 H), 7.70 (s, 1 H), 4.46 (s, 2 H), 3.51 - 3.63 (m, 2 H), 3.25 - 3.30 (m, 2 H), 2.15 - 2.28 (m, 2 H), 1.99 - 2.11 (m, 2 H)
Example 24
5-chloro-2-hydroxy-3-(((2-(2 -methoxy ethoxy)ethyl)(methyl)amino)methyl)-JV-(6- (trifluoromethyl)benzo[t/]thiazol-2-yl)benzamide (24) and 5-chloro-2-hydroxy-3-(((2-(2-
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SUBSTITUTE SHEET ( RULE 26 ) methoxy ethoxy)ethyl)(methyl)amino)methyl)-7V-(6-(trifluoromethyl)benzo[< ]thi azol -2- yl)benzamide hydrochloride (24A)
44.21 pL), 5-chloro-3-formyl-2-hydroxy-N-(6-(trifluoromethyl)benzo[<7]thiazol-2- yl)benzamide (150 mg, 374.29 pmol) in toluene (3 mL) was added acetic acid (22 mg, 374.29 pmol, 21.41 pL) to pH ~ 5-6. The mixture was stirred at 80 °C for 12 h. The reaction mixture was concentrated to remove solvent then MeOH (3 mL), and NaBHsCN (141 mg, 2.25 mmol) were added. The mixture was stirred at 20 °C for another 12 h, then (HCHO)n
(101 mg, 1.12 mmol) was added and the mixture was stirred at 20 °C for another 12 h The reaction mixture was concentrated to remove solvent, then purified by prep-HPLC (HC1 condition) to afford 5-chloro-2-hydroxy-3-(((2-(2- methoxyethoxy)ethyl)(methyl)amino)methyl)-A-(6-(trifluoromethyl)benzo[t/]thiazol-2- yl)benzamide (55.74 mg, 100.01 pmol, 26.72% yield, 99.478% purity, HC1) as a white solid. MS (ESI) [M+H]+ requires m/z 518.1, found m/z 518.2. XHNMR (400 MHz, METHANOL- d4) 5 ppm 8.20 - 8.30 (m, 2 H), 7.74 - 7.83 (m, 2 H), 7.71 (br d, J = 2.21 Hz, 1 H), 4.62 (br d, J = 11.25 Hz, 1 H), 4.35 (br d, J = 12.57 Hz, 1 H), 3.90 (br d, J = 4.41 Hz, 2 H), 3.67 - 3.75 (m, 2 H), 3.58 - 3.65 (m, 2 H), 3.51 (br s, 1 H), 3.44 (br s, 1 H), 3.41 (s, 3 H), 2.92 (s, 3 H).
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SUBSTITUTE SHEET ( RULE 26 ) Example 25
3-((bis(2 -methoxy ethyl)amino)methyl)-5-chloro-2 -hydroxy -N-(6- (trifluoromethyl)benzo[t/]thiazol-2-yl)benzamide (25) and 3-((bis(2- methoxyethyl)amino)methyl)-5-chloro-2-hydroxy-A-(6-(trifluoromethyl)benzo[t ]thiazol-2- yl)benzamide hydrochloride (25A)
[0423] To a solution of bis(2-methoxyethyl)amine (40 mg, 299.43 pmol, 44.21 pL) and 5-chloro-3-formyl-2-hydroxy-A-(6-(trifluoromethyl)benzo[</]thiazol-2-yl)benzamide (100 mg, 249.53 pmol, Example 22) in toluene (3 mL) was added acetic acid (3 mg, 49.91 pmol, 2.85 pL) to pH ~ 5-6. The mixture was stirred at 75 °C for 12 h. The reaction mixture was concentrated to remove solvent, then MeOH (3 mL) and NaBHsCN (94 mg, 1.5 mmol) were added. The mixture was stirred at 20 °C for another 5h The reaction mixture was concentrated to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 3-((bis(2-methoxyethyl)amino)methyl)-5-chloro-2-hydroxy-A-(6- (trifluoromethyl)benzo[c/]thiazol-2-yl)benzamide (98.93% purity, HC1) 47.28 mg as a brown solid. MS (ESI) [M+H]+ requires m/z 518.1, found m/z 518.1. 'H NMR (400 MHz, METHANOL-O 5 ppm 8.26 (s, 1 H), 8.15 (d, J = 2.44 Hz, 1 H), 7.73 - 7.80 (m, 2 H), 7.56 (d, J= 2.45 Hz, 1 H), 4.52 (s, 2 H), 3.74 - 3.80 (m, 4 H), 3.44 - 3 50 (m, 4 H), 3.41 (s, 6 H).
[0424] Compounds 26-37 were made in a similar manner as Compound 25 in Example 25:
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SUBSTITUTE SHEET ( RULE 26 ) Example 26
5-chloro-2-hydroxy-3-(piperidin- l-ylmethyl)-/V-(6-(trifluoromethyl)benzo[t/]thiazol-2- yl)benzamide (26)
[0425| MS (ESI) [M+H]+ requires m/z 470.08, found m/z 470.0. 'H NMR (400 MHz, METHANOL-d4) 5 ppm 8.24 - 8.29 (m, 2 H), 7.74 - 7.82 (m, 2 H), 7.68 (d, J = 2.45 Hz, 1 H), 4.36 (s, 2 H), 3.46 - 3.59 (m, 2 H), 3.02 - 3.18 (m, 2 H), 1.96 (br d, J = 14.18 Hz, 2 H), 1.77 (br d, J = 13.69 Hz, 3 H), 1.56 (br s, 1 H).
Example 27
3-(azetidin-l-ylmethyl)-5-chloro-2-hydroxy-A-(6-(trifluoromethyl)benzo[t/]thiazol-2- yl)benzamide (27)
10426] MS (ESI) [M+H]+ requires m/z 442.85, found m/z 442.1. 'H NMR (400 MHz, METHANOL-c/4) 5 ppm 8.28 (s, 1 H) 8.24 (d, J=2.65 Hz, 1 H) 7.74 - 7.84 (m, 2 H) 7.65 (d, J=2.21 Hz, 1 H)
Example 28
5-chloro-2-hydroxy-3-((4-methylpiperazin-l-yl)methyl)-A-(6- (trifluoromethy l)b enzo[t/J th i azol -2-y l)b enzami de (28)
[0427] MS (ESI) [M+H]+ requires m/z 485.09, found m/z 485.0. 'H NMR (400 MHz, METI IANOL-<74) 8 ppm 8.27 (s, 1 H), 8.25 (d, J = 2.93 Hz, 1 H), 7.74 - 7.82 (m, 3 H), 4.50 (s, 2 H), 3.43 - 3.88 (m, 8 H), 3.02 (s, 3 H)
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SUBSTITUTE SHEET ( RULE 26 ) Example 29
5-chloro-2-hydroxy-3-((3-oxopiperazin-l-yl)methyl)-/V-(6-(trifluoromethyl)benzo[</]thiazol- 2-yl)benzamide (29)
[0428| MS (ESI) [M+H]+ requires m/z 485.06, found m/z 485.0. 'H NMR (400 MHz, METHANOL-J4) 5 ppm 8.28 - 8.31 (m, 2 H), 7.76 - 7.84 (m, 2 H), 7.72 (d, J = 2.63 Hz, 1 H), 4.53 (s, 2 H), 3.95 (s, 2 H), 3.60 (br s, 4 H)
Example 30
5-chloro-2-hydroxy-3-((3-hydroxypyrrolidin-l-yl)methyl)-A-(6-
(trifluoromethyl)benzo[c/]thiazol-2-yl (benzamide (30)
[0429] MS (ESI) [M+H]+ requires m/z 472.06 found m/z 472.0. 1 H NMR (400
MHz, DMSO-676) 5 ppm 8.41 (s, 1 H), 8.05 (d, J= 2.87 Hz, 1 H), 7.70 - 7.85 (m, 3 H), 4.43 (br d, J= 2.20 Hz, 1 H), 4.39 (s, 2 H), 3.41 - 3.50 (m, 2 H), 3.32 - 3.40 (m, 1 H), 3.15.
Example 31
5-chloro-2-hydroxy-3-((3 -hydroxy-3 -methylpyrrolidin- l -yl)mcthyl)-Af-(6- (trifluoromethyl(benzo[c/]thiazol-2-yl)benzamide (31)
|0430] MS (ESI) [M+H]+ requires m/z 486.08 found m/z 486.0. LH NMR (400 MHz, METHANOL-d4) 8 ppm 8.23 (br d, J = 16.63 Hz, 2 H), 7.77 (q, J = 8.64 Hz, 2 H), 7.69 (s, 1 H), 4.41 - 4.59 (m, 2 H), 3.47 - 3 87 (m, 2 H), 3.34 - 3.45 (m, 1.5 H), 3.22 (d, J = 11.74 Hz, 0.5 H), 1.97 - 2.30 (m, 2 H), 1.46 (s, 3 H).
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SUBSTITUTE SHEET ( RULE 26 ) Example 32
5-chloro-2-hydroxy-3-((3-methoxypyrrolidin-l-yl)methyl)-A-(6-
(trifluoromethy l)b cnzo[t/] th i azol -2-y l)b enzami de (32)
[04311 MS (ESI) [M+H]+ requires m/z 486.08 found m/z 486.1. LH NMR (400 MHz, METHANOL-d4) 8 ppm 8.19 - 8.28 (m, 2 H), 7.77 (q, J = 8.53 Hz, 2 H), 7.69 (br s, 1 H), 4.43 - 4.55 (m, 2 H), 4.16 - 4.24 (m, 1 H), 3.56 - 3.74 (m, 2 H), 3.32 - 3.50 (m, 5 H), 2.30 - 2.48 (m, 1 H), 2.06 - 2.21 (m, 1 H).
Example 33
3-((3-acetamidopyrrolidin-l-yl)methyl)-5-chloro-2-hydroxy-A-(6-
(trifluoromethy l)b enzo[t/J th i azol -2-y l)b enzami de (33 )
[0432] MS (ESI) [M+H]+ requires m/z 513.09 found m/z 513.0. 1HNMR (400 MHz, DMSO-d6) 8 ppm 8.45 (s, 1 H) 8.24 (br d, J=5.07 Hz, 1 H) 8.11 (d, J=2.65 Hz, 1 H)
7.84 - 7.91 (m, 2 H) 7.77 - 7.83 (m, 1 H) 4.44 (s, 2 H) 4.40 (br d, J=6.39 Hz, 1 H) 3.47 - 3.63 (m, 2 H) 3.32 - 3.44 (m, 1 H) 3.18 - 3.27 (m, 1 H) 2.27 - 2.39 (m, 1 H) 1.89 - 2.01 (m, 1 H)
1.84 (s, 3 H)
Example 34 l-(5-chloro-2-hydroxy-3-((6-(trifluoromethyl)benzo|X]thiazol-2- yl)carbamoyl)benzyl)pyrrolidine-2-carboxamide (34)
[0433[ MS (ESI) [M+H]+ requires m/z 499.07 found m/z 498.9. LH NMR (400
MHz, METHANOL-t74) 8 ppm 8.28 (s, 1 H), 8.23 (d, J = 2.65 Hz, 1 H), 7.74 - 7.84 (m, 2 H),
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SUBSTITUTE SHEET ( RULE 26 ) 7.67 (d, J= 2.43 Hz, 1 H), 4.45 - 4.58 (m, 2 H), 4.33 (dd, J= 9.37, 6.73 Hz, 1 H), 3.73 (br s, 1 H), 3.42 - 3.50 (m, 2 H), 2.51 - 2.67 (m, 1 H), 1.99 - 2.28 (m, 2 H)
Example (35)
5-chloro-3-((l,l-dioxidothiomorpholino)methyl)-2-hydroxy-A-(6- (trifluoromethyl)benzo[c/]thiazol-2-yl (benzamide (35)
[0434] MS (ESI) [M+H]+ requires m/z 520.03 found m/z 520.0. LH NMR (400
MHz, METHANOL-c/4) 5 ppm 8.27 - 8.31 (m, 2 H), 7.73 - 7.84 (m, 3 H), 4.59 (s, 2 H), 3.91
(br d, J= 5.73 Hz, 4 H), 3.57 (br s, 4 H)
Example 36
5-chloro-3-((3-cyanopyrrolidin-l-yl)methyl)-2-hydroxy-A-(6- (trifluoromethy l)b enzo[t/J th i azol -2-y l)b enzami de (36)
[0435| MS (ESI) [M+H]+ requires m/z 481.06 found m/z 480.9. 'H NMR (400 MHz, DMSO-d6) 5 ppm 8.42 (s, 1 H) 7.78 - 7.86 (m, 2 H) 7.69 (br d, J=8.38 Hz, 1 H) 7.38 (d, J=2.87 Hz, 1 H) 4.25 (br s, 2 H) 3.62 (br s, 1 H) 3.49 - 3.56 (m, 4 H) 2.16 - 2.36 (m, 2 H)
Example 37
5-chloro-2-hydroxy-3-((3-(methylsulfonyl)pyrrolidin-l-yl)methyl)-/V-(6- (trifluoromethy l)b enzo[<7] th i azol -2-y l)b enzami de (37)
[0436] MS (ESI) [M+H]+ requires m/z 534.05 found m/z 534.0. 1H NMR (400
MHz, METHANOL-J4) 6 ppm 8.22 - 8.30 (m, 2 H), 7.74 - 7.84 (m, 2 H), 7.70 (br d, J= 2.45
SUBSTITUTE SHEET ( RULE 26 ) Hz, 1 H), 4.51 - 4.58 (m, 2 H), 4.16 - 4.27 (m, 1 H), 3.83 - 3.99 (m, 2 H), 3.60 (br s, 2 H),
3.10 (s, 3 H), 2.50 - 2.69 (m, 2 H)
Example 38
(E)-5-chloro-2-hydroxy-3-((hydroxyimino)methyl)-7V-(6-(trifluoromethyl)benzo[t7]thiazol-2- yl)benzamide (38)
[0437] To a solution of 5-chloro-3-formyl-2-hydroxy-JV-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (100 mg, 249.53 pmol) in EtOH (3 mL) was added NH2OH.HCI (17.34 mg, 249.53 pmol) and Na2CO3 (52.89 mg, 499.05 pmol). The mixture was stirred at 20 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (basic condition) to afford the title compound (E)-5-chloro-2-hydroxy-3-((hydroxyimino)methyl)-7V- (6-(trifluoromethyl)benzo[tZ]thiazol-2-yl)benzamide (30.95 mg, 70.19 pmol, 28.13% yield, 94.297% purity) as a yellow solid. 'H NMR (400 MHz, METHANOL-d4) 5 ppm 8.38 (br s, 1 H), 8.30 (br s, 1 H), 8.05 (br s, 1 H), 7.87 (br s, 1 H), 7.72 (br d, J = 16.63 Hz, 2 H). MS (ESI) [M+H]+ requires m/z 416.00, found m/z 416.00.
Example 39
(£)-5-chloro-2-hydroxy-3-((methoxyimino)methyl)-AL(6-(trifluoromethyl)benzo[<i]thiazol-2- yl)benzamide (39)
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SUBSTITUTE SHEET ( RULE 26 ) [0438] (E)-5-chl oro-2 -hy droxy-3-((methoxyimino)methyl)-A-(6-
(trifluoromethyl)benzo[d]thiazol-2-yl)benzamide was made in a similar manner as compound 38. MS (ESI) [M+H]+ requires 430 02m/z found m/z 430.0. JH NMR (400 MHz, DMSO-de) 6 ppm 8.52 (br s, 1 H), 8.41 (br s, 1 H), 8.01 (br s, 1 H), 7.83 (br s, 2 H), 7.76 (br d, J = 0.98 Hz, 1 H), 3.94 (br s, 3 H).
Example 40
3-(acetamidomethyl)-5-chloro-2-hydroxy- V-(6-(trifluoromethyl)benzo[</]thiazol-2- yl)benzamide (40)
|0439j To a solution of (E)-5-chloro-2-hydroxy-3-((hydroxyimino)methyl)-A-(6- (trifluoromethyl)benzo[ ]thiazol-2-yl)benzamide (320 mg, 769.65 pmol, Example 40) in TFA (6 mL) was added Zn (251 mg, 3.85 mmol). The mixture was stirred at 25 °C for 12 h. The reaction mixture was concentrated to remove solvent. The residue was diluted with water (3 mL) and adjusted to pH 9 with 2M NaOH, then extracted with ethyl acetate (5 mL x 3). The combined organic layer was washed with brine (15 mL), dried over N 2SC>4, fdtered and concentrated under reduced pressure to afford 3-(aminomethyl)-5-chloro-2-hydroxy-2V- (6-(trifluoromethyl)benzo[t/]thiazol-2-yl)benzamide (300 mg, crude) as a white solid.
[0440] 3-(aminomethyl)-5-chloro-2-hydroxy-A-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (20 mg, 49.78 pmol) and TEA (6 mg, 59.73 pmol, 8.31 pL) was dissolved in DCM (3 mL) at 0 °C. The solution wasstirred and acetyl chloride (3 mg, 39.82 pmol, 2.84 pL) was added. The resulting solution was warmed to 20 °C for 30 minutes. The reaction mixture was quenched by addition of MeOH (3 mL), then concentrated under reduced pressure to give a residue. The residue was purified by prep- HPLC (HC1 condition) to afford 3-(acetamidomethyl)-5-chloro-2-hydroxy-Ar-(6-
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SUBSTITUTE SHEET ( RULE 26 ) (trifluoromethyl)benzo[c/]thiazol-2-yl)benzarriide (14 mg, 99.660% purity) as a white solid. XH NMR (400 MHz, METHANOL-^) 5 ppm 8.28 (s, 1 H), 8.03 (d, J= 2.45 Hz, 1 H), 7.73 - 7.86 (m, 2 H), 7.43 (s, 1 H), 4.39 (s, 2 H), 2.03 (s, 3 H) MS (ESI) [M+H]+ requires m/z 444.03, found m/z 444.0.
Example 41
5-chloro-2-hydroxy-JV1,7V1-bis(2-methoxyethyl)-JV3-(6-(trifluoromethyl)benzo[t/]thiazol-2- yl)isophthalamide (41)
SUBSTITUTE SHEET ( RULE 26 ) [04411 To a flame dried flask was added NBS (2.99g, 16.79 mmol), AIBN (173 mg, 1.05 mmol) and a solution of methyl 5-chloro-2-methoxy-3-methylbenzoate (1.13g, 5.26 mmol) in CC14 (20 mL). The suspension was refluxed in the dark overnight The mixture was cooled to rt and concentrated. The residue was purified via silica gel column chromatography to give methyl 5-chloro-3-(dibromomethyl)-2-methoxybenzoate as a yellow oil (1.59g, 81%). 'H NMR (300 MHz, cdcl3) 6 8 04 (d, J = 2.7 Hz, 1H), 7 80 (d, J = 2.7 Hz, 1H), 7 09 (s, 1H), 3.95 (s, 6H).
[0442] Methyl 5-chloro-3-(dibromomethyl)-2-methoxybenzoate (1.59g, 4.27 mmol) was dissolved in 10 mL concentrated sulfuric acid and the mixture was stirred at rt for 2h. The reaction mixture was poured into ice water and extracted with ethyl acetate two times. The combined organic layer was washed with brine and dried over sodium sulfate. After concentration, the residue was purified via silica gel column chromatography to give methyl 5-chloro-3-formyl-2 -methoxybenzoate (673 mg, 71%) as a white solid. 'H NMR (300 MHz, cdcl3) 5 10.33 (s, 1H), 8.00 (d, J = 2.6 Hz, 1H), 7.91 (d, J = 3.1 Hz, 1H), 3.98 (s, 4H), 3.94 (s, 4H).
[0443] To a stirred solution of methyl 5-chloro-3-formyl-2-methoxybenzoate (112 mg, 0.49 mmol) in acetone (3 mL) and water (3 mL) was added KMnCU (310 mg, 1.96 mmol). The resulting mixture was stirred at rt for 3.5h. After completion of the reaction as indicated by TLC, 0.8g sodium carbonate was added and stirred for another 15minutes. The mixture was then filtered and the pH of the filtrate was adjusted to 2-3 when a large amount of solid was precipitated. The mixture was filtered and the solid was washed with water. The crude product was purified through recrystallization in ethanol to give 5-chloro-2-methoxy-3- (methoxycarbonyl)benzoic acid (90 mg, 76%) as a white solid. MS (ESI) [M+H]+ requires m/z 245.02, found m/z 245.0.
[0444] 5-chloro-2-methoxy-3-(methoxycarbonyl)benzoic acid (90 mg, 0.37 mmol) was dissolved in DCM (3.0 mL), followed by the addition of a catalytic amount of DMF (1 drop) and oxalyl chloride (38 pL, 0.44 mmol) respectively. The reaction was allowed to stir at rt for 30 minutes and concentrated in vacuo. The residue was re-dissolved in THF (5.0 mL), and Hunig’s base (192 pL, 1.1 mmol) and bis(2-methoxyethyl)amine (55 pL, 0.37 mmol) were added. The mixture was stirred at rt for 48 hours before silica gel was added to quench the reaction. The solvent was evaporated and the resulting residue was purified via silica gel column chromatography to yield methyl 3 -(bi s(2 -methoxy ethyl)carbam oyl)-5- chloro-2-methoxybenzoate (116 mg, 88%) as a colorless oil. 'H NMR (300 MHz, cdcl3) 5 7.81 (d, J= Hz, 1H), 7.42 (d, J= 2.7 Hz, 1H), 4.04 (M, 1H), 3.93 (s, 3H), 3.88 (s, 3H),
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SUBSTITUTE SHEET ( RULE 26 ) 3.71 - 3.62 (m, 2H), 3.57 - 3.40 (m, 3H), 3.39 (s, 3H), 3.37 - 3.24 (m, 2H), 3.23 (s, 3H). MS (ESI) [M+H]+ requires m/z 360.12, found m/z 360.10.
|0445] To a stirred solution of methyl 3-(bis(2-methoxyethyl)carbamoyl)-5- chloro-2-methoxybenzoate (116 mg, 0.323 mmol) in MeOH (3 mL) was added 2.0 m IN KOH solution. The resulting mixture was stirred at rt overnight. The solvent was evaporated out and the residue was portioned between ethyl acetate and IN NaOH solution. The aqueous layer was acidified to pH=l and extracted with ethyl acetate. This organic phase was dried over over sodium sulfate and concentrated in vacuo to afford 3-(bis(2- methoxyethyl)carbamoyl)-5-chloro-2-methoxybenzoic acid (115 mg, 100%) as a colorless oil which used in the next step without further purification. MS (ESI) [M+H]+ requires m/z 346.11, found m/z 346.20.
[0446] 3-(bis(2-methoxyethyl)carbamoyl)-5-chloro-2-methoxybenzoic acid (115 mg, 0.32 mmol) was dissolved in DCM (3.0 mL), followed by the addition of a catalytic amount of DMF (1 drop) and oxalyl chloride (34 pL, 0.39 mmol) respectively. The reaction was allowed to stir at rt for 30 minutes and concentrated in vacuo. The residue was redissolved in THF (5.0 mL), and Hunig’s base (170 pL, 0.98 mmol) and 6- (trifluoromethyl)benzo[d]thiazol-2-amine (71 mg, 0.32 mmol) were added. The mixture was stirred at rt for 48 hours before silica gel was added to quench the reaction. The solvent was evaporated and the resulting residue was purified via silica gel column chromatography to yield 5-chloro-2-methoxy-A1,A1-bis(2-methoxyethyl)-/V3-(6-(trifluoromethyl)benzo[6 ]thiazol- 2-yl)isophthalamide (95 mg, 58%) as a colorless oil. MS (ESI) [M+H]+ requires m/z 546.11, found m/z 546.20.
[0447] A solution of 5-chloro-2-methoxy-A1,A1-bis(2-methoxyethyl)-/V3-(6- (trifluoromethyl)benzo[t/]thiazol-2-yl)isophthalamide (95 mg, 0.174 mmol) in DMF (3 mL) was mixed with sodium ethanethiolate (74 mg, 0.872 mmol) and the resulting suspension was heated at 130 °C overnight. After completion of the reaction, IN HC1 was added to the reaction and extracted with ethyl acetate two times. The combined ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give 5-chloro-2-hydroxy-A1,A1-bis(2- methoxyethyl)-A3-(6-(trifluoromethyl)benzo[ ]thiazol-2-yl)isophthalamide (55 mg, 63%) as a yellow solid. XHNMR (300 MHz, cdcl3) 5 8.18 - 7.95 (m, 5H), 3.71 (brs, 4H), 3.56 (brs, 4H), 2.95 (s, 3H), 2.87 (s, 3H). MS (ESI) [M+H]+ requires m/z 532.09, found m/z 532.20.
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SUBSTITUTE SHEET ( RULE 26 ) Example 42
5-chloro-2V-(6-(difluoromethyl)benzo[<i]thiazol-2-yl)-2-hydroxy-3-((2- methoxyethoxy)methyl)benzamide (42)
[0448] To a solution of ethyl 2-anii nobeiizo[c/]tlii azol e-6-carboxyl ale (3 g, 13.50 mmol) in THF (60 mL) was added portion wise DIBAL-H (1 M, 53.99 mb) at 0 °C. The reaction mixture was stirred at 50 °C for 12 h. The reaction mixture was quenched by addition saturated sodium potassium tartrate (30 mL), and then diluted with water (100 mL) and extracted with ethyl acetate (200 mL x 3). The combined organic layer was washed with brine (200 mL), dried over NazSO-i, filtered and concentrated under reduced pressure to give (2-aminobenzo[</]thiazol-6-yl)methanol (2.4 g, crude) as a yellow solid. ’H NMR (400 MHz, DMSO-6/6) 8 ppm 7.57 (s, 1 H), 7.38 (s, 2 H), 7.27 (d, J= 8.31 Hz, 1 H), 7.14 (dd, J= 8.07,
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SUBSTITUTE SHEET ( RULE 26 ) 1.22 Hz, 1 H), 5.09 (t, J= 5.87 Hz, 1 H), 4.47 (d, J= 5.87 Hz, 2 H). MS (ESI) [M+H]+ requires m/z 181.04, found m/z 181.2.
|0449] To a solution of (2-aminobenzo[< ]thiazol-6-yl)methanol (1 g, 5.55 mmol) in ethyl acetate (30 mL) was added MnCb (4.8 g, 55.49 mmol). The mixture was stirred at 25 °C for 12 h. The reaction mixture was filtered with diatomite filter and concentrated under reduced pressure to give the crude product 2-aminobenzo[d]thiazole-6-carbaldehyde (900 mg, crude) as a yellow solid. 'H NMR (400 MHz, DMSO- e) 5 ppm 9.87 (s, 1 H), 8.23 (d, J = 1.47 Hz, 1 H), 8.05 (br s, 2 H), 7.75 (dd, J= 8.31, 1.47 Hz, 1 H), 7.44 (d, J= 8.31 Hz, 1 H). MS (ESI) [M+H]+ requires m/z 179.02, found m/z 179.1.
[0450] To a stirred solution of 5-chloro-2-methoxybenzoic acid (10 g, 53.59 mmol) in H2SO4 (18.25 mL, purity 98%) and TFA (36.50 mL) at 20 °C was added NBS (10.5 g, 58.95 mmol). The pale solution was stirred at 20 °C for 12 h. The resulting pale suspension was carefully poured onto crushed ice (100 mL). The mixture was extracted with ethyl acetate (50 mL x 3), dried over NazSCU and concentrated under reduced pressure. The light yellow residue was suspend in DCM (50 mL). The solid was collected, washed with cold DCM (50 mL) and dried under vacuum to afford 3-bromo-5-chloro-2 -methoxybenzoic acid (10 g, crude) as a white solid. 'H NMR (400 MHz, DMSO-de) 5 ppm 7.96 (t, J = 2.32 Hz, 1 H), 7.69 (t, I = 2.32 Hz, 1 H), 3.78 (d, J = 2.43 Hz, 3 H)
[0451] To a stirred solution of 3 -bromo-5-chloro-2 -methoxybenzoic acid (5 g, 18.83 mmol) in DMF (25 mL) was added K2CO3 (26 g, 188.33 mmol) followed by CH3I (2.7g, 18.83 mmol, 1.2 mL). The mixture was stirred at 20 °C for 12 h. H2O (50 mL) was added and extracted with ethyl acetate (30 mL x 2). The combined organic layer was washed with brine (100 mL) and dried over sodium sulfate. The organic layer was filtered and the solvent removed in vacuo to afford methyl 3-bromo-5-chloro -2 -methoxybenzoate (4.7 g, purity 88.4%) as yellow oil. 'H NMR (400 MHz, DMSO-d6) 5 ppm 8.03 (d, J = 2.93 Hz, 1 H), 7.74 (d, J = 2.45 Hz, 1 H), 3.87 (s, 3 H), 3.81 (s, 3 H). MS (ESI) [M+H]+ requires m/z 278.93, found m/z 281.0.
[0452] A mixture of methyl 3-bromo-5-chloro-2 -methoxybenzoate (2 g, 7.16 mmol), potassium trifluoro (methyl)boranuide (1 g, 8.59 mmol), K2CO3 (3 g, 21.47 mmol), Pd(dppf)C12 (1 g, 1.43 mmol) in dioxane (20 mL), H2O (2 mL) was degassed and purged with N2 3 times, and then the mixture was stirred at 90 °C for 12 h under N2 atmosphere. The reaction mixture was concentrated to remove solvent. The residue was purified by column chromatography (SiCh, petroleum ether/ethyl acetate = 1/0 to 10/1) to afford methyl 5-chloro- 2-methoxy-3-methyl-benzoate (1.2 g, 5.31 mmol, 74.23% yield, 95% purity) as yellow oil.
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SUBSTITUTE SHEET ( RULE 26 ) [04531 To a flame dried flask was added NBS (1.2 g, 6.71 mmol), AIBN (91 mg, 559.06 pmol) and a solution of methyl 5-chloro-2-methoxy-3-methyl-benzoate (1.2 g, 5.59 mmol) in CCL (10 mL). The suspension was stirred at 80 °C for 12 h. The reaction mixture was concentrated to remove solvent. The residue was purified by column chromatography (SiCh, petroleum ether/ethyl acetate = 1/0 to 100/1) to afford methyl 3-(bromomethyl)-5- chloro-2-methoxybenzoate (1.5 g, 3.58 mmol, 63.98% yield, 70% purity) as yellow oil XH NMR (400 MHz, CHLOROFORM-d) 5 ppm 7.69 (d, J = 2.93 Hz, 1 H), 7.46 (d, J = 2.93 Hz, 1 H), 4.44 (s, 2 H), 3.88 (s, 3 H), 3.86 (s, 3 H).
[0454| To a stirred solution of methyl 3-(bromomethyl)-5-chloro-2- methoxybenzoate (1 g, 3.41 mmol) in 2-methoxyethanol (19.3 g, 253.63 mmol, 20 mL) was added 2NNaOH (15 mL). The resulting mixture was stirred at 75 °C for 12 h then concentrated in vacuo. The residue was diluted with water (5 mL) and adjusted to pH 9 with 2M NaOH, then extracted with ethyl acetate (5 mL x 3). The clear solution was acidified with IN HC1 to pH 5 and then extracted with ethyl acetate (5 mL x 3). The combined organic layer was washed with brine (20 mL) and dried over Na2 Ch, filtered and concentrated under reduced pressure to afford 5-chloro-2-methoxy-3-((2-methoxyethoxy)methyl)benzoic acid (500 mg, purity:80%) as yellow oil. 'H NMR (400 MHz, CHLOROFORM-d) 6 ppm 7.90 - 8.00 (m, 1 H), 7.63 - 7.73 (m, 1 H), 4.63 (s, 2 H), 3.92 - 3.94 (m, 3 H), 3.72 (dd, J = 5.62, 3.67 Hz, 2 H), 3.59 - 3.65 (m, 2 H), 3.42 (s, 3 H). [M-H]’ requires m/z 273.0, found m/z 272.9.
[0455] To a solution of 2-aminobenzo[r/]thiazole-6-carbaldehyde (162 mg, 910.09 pmol), 5-chloro-2-methoxy-3-((2-methoxyethoxy)methyl)benzoic acid (250 mg, 910.09 pmol) in DCM (4 mL) was added EDCI (209.36 mg, 1.09 mmol) and HOBt (184 mg, 1.37 mmol). The mixture was stirred at 50 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (neutral condition) to afford 5-chloro-A-(6-formylbenzo[ ]thiazol-2-yl) -2 -m ethoxy-3 -((2- methoxyethoxy)methyl)benzamide (80 mg, 97% purity) as a yellow solid. [M+H]+ requires m/z 435.07, found m/z 435.1
[0456| To a solution of 5-chloro-JV-(6-formylbenzo[t/]thiazol-2-yl) -2-methoxy-3- ((2 -methoxyethoxy )methyl)benzamide (50 mg, 114.97 pmol) in DCM (2 mL) was added BAST (406 mg, 1.84 mmol, 402.95 pL). The mixture was stirred at 50 °C for another 12 h. The reaction mixture was quenched by addition sat.NaHCCL (8 mL), and extracted with ethyl acetate (8 mL x 3). The combined organic layer was washed with brine (10 mL), dried over NazSCh, filtered and concentrated under reduced pressure to give a residue. The residue was
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SUBSTITUTE SHEET ( RULE 26 ) purified by prep-TLC (SiC>2, petroleum ether/ethyl acetate = 3: 1) to afford 5-chloro-/V-(6- (difluoromethyl)benzo[d]thiazol-2-yl)-2-methoxy-3-((2-methoxyethoxy)methyl)benzamide (40 mg, 72.67 pmol, 63.20% yield, 83% purity) as a white solid. [M+H]+ requires m/z 457.07, found m/z 457.2
[0457] To a solution of 5-chloro-/V-(6-(difluoromethyl)benzo[d]thiazol-2-yl)-2- methoxy-3-((2-methoxyethoxy)methyl)benzamide (40 mg, 87.55 pmol) in DMF (2 mL) was added ethyl sulfanyl sodium (22 mg, 262.65 pmol). The mixture was stirred at 160 °C for 3 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with water (3 mL) and extracted with DCM (3 mL x 3). The combined organic layers were washed with brine (5 mL), dried over NaiSCU, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCh, Petroleum ether/Ethyl acetate = 0:1). The residue was purified by prep-HPLC (HC1 condition) to afford 5-chloro-A'-(6-(difluorornethyl)benzo[<7]thiazol-2-yl)-2-hydroxy-3-((2- methoxyethoxy)methyl)benzamide (5.22 mg, 99.157% purity) was obtained as a white solid. 1 H NMR (400 MHz, DMSO-d6) 5 ppm 8.28 (s, 1 H), 7.99 (br s, 1 H), 7.76 (br s, 1 H), 7.67 - 7.73 (m, 1 H), 7.52 (s, 1 H), 6.98 - 7.33 (m, 1 H), 4.56 (s, 2 H), 3.65 (dd, J = 5.62, 3.67 Hz, 2 H), 3.52 (dd, J = 5.62, 3.67 Hz, 2 H), 3.28 (s, 3 H). [M+H]+ requires m/z 443.06, found m/z 442.9
Example 43
5-fluoro-2-hydroxy-3-((2-methoxyethoxy)methyl)-/V-(6-(trifluoromethylbenzo[<7]thiazol-2- yl)benzamide (43)
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SUBSTITUTE SHEET ( RULE 26 )
[0458] To a stirred solution of 5-fluoro-2 -methoxybenzoic acid (5 g, 29.39 mmol) in H2SO4 (10 mL) (98%) and TFA (20 mL) at 20°C was added NBS (5.7 g, 32.33 mmol). The pale solution was stirred at 20°C for 12 h. The resulting pale suspension was carefully poured onto crushed ice (500 mL). The mixture was extracted with ethyl acetate (300 mL x 3). The combined organic layer was washed with brine 1000 mL, dried over Na2SC>4, filtered and concentrated under reduced pressure to afford 3-bromo-5-fluoro-2 -methoxybenzoic acid (7 g) as a yellow solid. 'H NMR (400 MHz, DMSO- r,) 5 ppm 7.77 (dd, J= 7.89, 3.07 Hz, 1 H), 7.50 (dd, 7= 8.77, 3.07 Hz, 1 H), 3.76 - 3.80 (s, 3 H).
(0459] To a stirred solution of 3 -bromo-5-fluoro-2-m ethoxybenzoic acid (7 g, 28.11 mmol) in DMF (20 mL) was added K2CO3 (38.8 g, 281.09 mmol) followed by Mel (3.9 g, 28.11 mmol, 1.75 mL). The mixture was stirred at 20°C for 12. Water (100 mL) was added and the extracted with ethyl acetate (150 mL x 3). The combined organic layer was washed with brine (200 mL) and dried over sodium sulfate. The organic layer was filtered and the solvent was removed in vacuo. The residue was purified by column chromatography (SiCh, petroleum ether/ethyl acetate 1/0 to 100/1) to afford methyl 3-bromo-5-fluoro-2- methoxybenzoate (4.6 g, 11.19 mmol, 39.81% yield, 64% purity) as colorless oil.
|0460] A mixture of methyl 3-bromo-5-fluoro-2 -methoxybenzoate (3 g, 11.40 mmol), potassium;trifluoro(methyl)boranuide (1.5 g, 12.54 mmol), K2CO3 (4.7 g, 34.21
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SUBSTITUTE SHEET ( RULE 26 ) mmol) and Pd(dppf)Ch (834 mg, 1.14 mmol) in dioxane (30 mL) and H2O (3 mL) was degassed and purged with N2 3 times, then the mixture was stirred at 90°C for 12 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by column chromatography (Si O2, petroleum ether/ethyl acetate 1/0 to 100/1) to afford methyl 5-fluoro-2-methoxy-3-methylbenzoate (1.8 g, 8.17 mmol, 71.67% yield, 90% purity) as yellow oil. 'H NMR (400 MHz, CHLOROFORM-*/) 5 ppm 7.31 (dd, J= 8.60, 2.87 Hz, 1 H), 7.01 - 7.06 (m, 1 H), 3.90 (s, 3 H), 3.78 (s, 3 H), 2.29 (s, 3 H)
[0461] Methyl 5-fluoro-2-methoxy-3-methylbenzoate (1.8 g, 9.08 mmol), NBS (1.2 g, 7.27 mmol) and AIBN (149 mg, 908.22 umol) in CCL (30 mL), were combined and the mixture was stirred at 80°C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by column chromatography (SiO2, petroleum ether/ethyl acetate 1/0 to 100/1) to afford methyl 3-(bromomethyl)-5-fluoro- 2 -methoxybenzoate (2.1 g, 6.06 mmol, 66.76% yield, 80% purity) as yellow oil. 'H NMR (400 MHz, CHLOROFORM-*/) 8 ppm 7.43 (dd, J= 8.31, 3.42 Hz, 1 H), 7.19 - 7.24 (m, 1 H), 4.46 (s, 2 H), 3.87 (s, 3 H), 3.86 (s, 3 H)
|0462] To methyl 3-(bromomethyl)-5-fluoro-2-methoxy-benzoate (2.1 g, 7.58 mmol) in NaOH (30 mL) (2M) was added 2-methoxyethanol (40.5 g, 532.63 mmol, 42.00 mL), the mixture was stirred at 75°C for 12 h. The reaction mixture extracted with ethyl acetate (40 mL x 3) The combined water layers were adjusted pH ~ 4 by addition of HC1 (2N), and then the mixture was extracted with ethyl acetate (50 mL x 3); the combined organic layer was concentrated under reduced pressure to afford 5-fluoro-2-methoxy-3-(2- methoxyethoxymethyl)benzoic acid (1.9 g, crude) as a yellow oil.
[0463| 5-fluoro-2-m ethoxy-3 -(2-methoxyethoxymethyl)benzoic acid (500 mg,
1.94 mmol), 6-(trifluoromethyl)benzo[*/]thiazol-2-amine (422 mg, 1.94 mmol), EDCI (445 mg, 2.32 mmol) and HOBt (392 mg, 2.90 mmol) in DCM (15 mL), the mixture was stirred at 50°C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (neutral condition) to afford 5-Fluoro-2- methoxy-3-((2 -methoxy ethoxy)methyl))- V-[6-(trifluoromethyl)benzo[*/]thiazol-2- yl]benzamide (300 mg, 523.53 umol, 27.04% yield, 80% purity) as a white solid.
|0464] 5 -Fluoro-2-methoxy-3-((2-methoxyethoxy)methyl))-7V-[6-
(trifluoromethyl)benzo[*/]thiazol-2-yl]benzamide (50 mg, 109.07 pmol) and NaSEt (45 mg, 545.34 pmol) in DMF (2 mL), the mixture was stirred at 160 °C for 1 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was
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SUBSTITUTE SHEET ( RULE 26 ) purified by prep-TLC (SiCh, petroleum ether/ethyl acetate = 1: 1) and then the residue was purified by prep-HPLC (HC1 condition) to afford 5-fluoro-2-hydroxy-3-((2- methoxyethoxy)methyl)-Ar-(6-(trifluoromethylbenzo[<7]thiazol-2-yl)benzamide (13.33 mg, 30.00 pmol, 27.51% yield, 100% purity) as a white solid. 1HNMR (400 MHz, METHANOL- c/4) 5 ppm 8.27 (s, 1 H), 7.71 - 7.85 (m, 3 H), 7.40 (br d, J= 6.85 Hz, 1 H), 4.65 (s, 2 H), 3.71 - 3 77 (m, 2 H), 3.63 (dd, J= 5.62, 3.67 Hz, 2 H), 3.41 (s, 3 H) MS (ESI) [M+H]+ requires m/z 445.0, found m/z 445.0
Example 44
5-chloro-2-hvdroxy-3-((2-methoxyethoxy)methyl)-N-(6-(trifluoromethyl)pyridin-3- yl)benzamide(44) and 5-chloro-2-hvdroxy-3-((2-methoxyethoxy)methyl )-N-f6- (trifluoromethyl)pyridin-3-yl)benzamide hydrochloride (44A)
[0465| To a solution of 5-chloro-2-methoxy-3-(2-methoxyethoxymethyl)benzoic acid (200 mg, 728.08 pmol), 6-(trifluoromethyl)pyridin-3-amine (118 mg, 728.08 pmol) in Py (2 mL) was added POCI3 (167 mg, 1.09 mmol, 101.49 pL) at 0°C. The mixture was stirred at 0°C for 1 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with water (3 mL) and extracted with DCM (3 mL x 3). The combined organic layer was washed with brine (5 mL), dried over NazSCk, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep- TLC (SiCh, petroleum ether/ethyl acetate 3/1) to afford 5-chloro-2-methoxy-3-((2- methoxyethoxy)methyl)-2V-(6-(trifluoromethyl)pyridine-3-yl)benzamide (117 mg, 212.32
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SUBSTITUTE SHEET ( RULE 26 ) pmol, 29.16% yield, 76% purity) as a white solid. MS (ESI) [M+H]+ requires m/z 419.09, found m/z 419.2
|0466] To 5-chloro-2-methoxy-3-((2-methoxy ethoxy )methyl)-A-(6- (trifluoromethyl)pyridine-3-yl)benzamide (95 mg, 226.84 pmol) in DMF (2 mL) was added NaSEt (38 mg, 453.68 umol) and the mixture was stirred at 160°C for 2 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 5-chloro-2-hydroxy-3-((2- methoxyethoxy)methyl)-N-(6-(trifluoromethyl)pyridin-3-yl)benzamide (32.99 mg, 73.67 pmol, 32.48% yield, 98.53% purity, HC1) as a brown solid. XH NMR (400 MHz, METHANOL-^) 8 ppm 8.93 (d, J= 2.19 Hz, 1 H), 8.37 (dd, J= 8.33, 2.19 Hz, 1 H), 7.90 (d, J= 2.19 Hz, 1 H), 7.74 (d, J = 8.77 Hz, 1 H), 7.52 (d, J = 2.63 Hz, 1 H), 4.53 (s, 2 H), 3.60 - 3.66 (m, 2 H), 3.51 - 3.56 (m, 2 H), 3.22 (dt, J= 3.29, 1.43 Hz, 3 H). MS (ESI) [M+H]+ requires m/z 405.08, found m/z 405.0
Example 45
5-chloro-2-hydroxy-3-((2 -methoxy ethoxy )methyl)-A-(5-(trifluoromethyl)pyri din-2- yl)benzamide (45)
[0467] 5-chl oro-2 -hydroxy-3 -((2-methoxy ethoxy)methyl)-A-(5-
(trifluoromethyl)pyridin-2-yl)benzamide was made in a manner similr to Example 44. LCMS: MS (ESI) [M+H]+ requires m/z 405.08, found m/z 405.0. 1HNMR (400 MHz, METHANOL-c/i) 8 ppm 8.69 (s, 1 H), 8.43 (d, J= 8.82 Hz, 1 H), 8.13 (dd, J= 8.93, 2.09 Hz, 1 H), 8.00 (d, J= 2.43 Hz, 1 H), 7.60 (d, J= 2.43 Hz, 1 H), 4.65 (s, 2 H), 3.71 - 3.77 (m, 2 H), 3.61 - 3.66 (m, 2 H), 3.41 (s, 3 H)
Example 46
2-hy droxy-5-i odo-3 -((2-methoxy ethoxy)methyl)-A-(6-(trifluoromethyl)benzo[<7]thiazol-2- yl)benzamide (46)
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SUBSTITUTE SHEET ( RULE 26 )
[0468| 5-Iodo-2-methoxy-3-((2-methoxyethoxy)methyl)-A-(6- (trifluoromethyl)benzo[4/]thiazol-2-yl)benzamide (Example 48; 40 mg, 70.63 pmol, 1 eq) in DMF (2 mL) was stirred at 160 °C for 10 minutes, then NaSEt (29 mg, 353.15 pmol) was added, the mixture was stirred at 160 °C for 1 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 2-hydroxy-5-iodo-3-((2-methoxyethoxy)methyl)-JV-(6- (trifluoromethyl)benzo[</]thiazol-2-yl)benzamide (10.26 mg, 18.55 pmol, 26.26% yield, 99.848% purity) as a white solid. 'H NMR (400 MHz, METHANOL-^) 3 ppm 8.33 (d, J = 1.96 Hz, 1 H), 8.26 (s, 1 H), 7.86 (s, 1 H), 7.77 (br t, J= 8.80 Hz, 2 H), 4.62 (s, 2 H), 3.72 (dd, J= 5.62, 3.67 Hz, 2 H), 3.62 (dd, J= 5.62, 3.67 Hz, 2 H), 3.39 - 3.42 (s, 3 H). MS (ESI) [M+H]+ requires m/z 552.9 , found m/z 553.0.
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SUBSTITUTE SHEET ( RULE 26 ) Example 47
5-chloro-A-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-hydroxy-3-((2- methoxyethoxy)methyl)benzamide (47)
10469] To a solution of 5-chloro-2-methoxy-3-((2-methoxyethoxy)methyl)benzoic acid (50 mg, 182.02 pmol, Example 2), 2-chloro-6-(trifluoromethyl)pyridin-3-amine (35 mg, 182.02 ymol) in pyridine (2 m ) was added POCh (41 mg, 273.03 pmol, 25.37 pL) at 0 °C. The mixture was stirred at 20 °C for 1 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-TLC (SiCh, petroleum ether/ethyl acetate = 2:1) to afford 5-chloro-A-(2-chloro-6-(trifluoromethyl)pyridine-3yl)-2- methoxy-3-((2-methoxyethoxy)methyl)benzamide (75 mg, 65% purity) as a white solid.
[0470] To a mixture of EtSH (1 mL) and DCM (1 mL) was added A1CL (66 mg, 496.43 pmol, 27.13 pL) at 0 °C, the resulting solution was warmed to 20 °C, and then 5- chloro-A-(2-chloro-6-(trifluoromethyl)pyridine-3yl)-2-methoxy-3-((2- methoxy ethoxy )methyl)benzamide (75 mg, 165.48 pmol) was added with stirring and the mixture was stirred at 20 °C for 9 hs. The mixture was poured into 2 mL of water, acidified with HC1 to pH ~ 4-5, then extracted with DCM (2 x 3), dried over Na2SOr, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep- HPLC (HC1 condition) to afford 5-chloro-A-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2- hydroxy-3-((2-methoxyethoxy)methyl)benzamide (29.94 mg, 62.94 pmol, 38.04% yield, 100% purity, HC1) as a white solid. LH NMR (400 MHz, METHANOL-^) 8 ppm 8.90 (br d, J= 8.31 Hz, 1 H), 8.02 (br s, 1 H), 7.85 (br d, J= 8.31 Hz, 1 H), 7.56 (br s, 1 H), 4.69 (s, 2
-210-
SUBSTITUTE SHEET ( RULE 26 ) H), 3.75 (br s, 2 H), 3.64 (br s, 2 H), 3.42 (s, 3 H). MS (ESI) [M+H]+ requires m/z 439.04, found m/z 439.0
Example 48 and 49
5-iodo-2-methoxy-3-((2-methoxyethoxy)methyl)-Af-(6-(trifluoromethylbenzo[d]thiazol-2- yl]benzamide (48) 2-hy droxy -3 -((2-m ethoxy ethoxy)methyl)-5-methyl-A-(6- (trifluoromethyl)benzo[c/]thiazol-2-yl (benzamide (49 A) 2-hydroxy-3-((2- methoxyethoxy)methyl)-/V-(6-(trifluoromethyl)benzo[t/]thiazol-2-yl)benzamide (49B)
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SUBSTITUTE SHEET ( RULE 26 )
[0471| I2 (50 g, 197.18 mmol, 39.7 mL) and KI (27.2 g, 164.31 mmol) in H2O (125 mL) were slowly added to a 0 °C mixture of 2 -hydroxy-3 -methylbenzoic acid (25 g, 164.31 mmol) in KOH (200 mL, 10%), and the mixture was stirred at 0 °C for 1 h. The reaction mixture was diluted with water (500 mL) and extracted with ethyl acetate (1000 mL x 3). The clear solution was acidified with IN HC1 to pH 5 and then extracted with ethyl acetate (1000 mL x 3). The combined organic layer was washed with brine (2000 mL), dried over Na2SO 1, filtered and concentrated under reduced pressure to afford 2-hydroxy-5-iodo-3- methylbenzoic acid (17 g, crude) as a yellow solid.
[0472| 2-hydroxy-5-i odo-3 -methylbenzoic acid (7 g, 25.18 mmol) and K2CCL (34.7 g, 251.76 mmol) in DMF (140 mL) was stirred at 20 °C for 30 minutes, then Mel (7.8 g, 55.39 mmol, 3.45 mL) was added dropwise, and the mixture was stirred at 20 °C for another 12 h. H2O (300 mL) was added and extracted with ethyl acetate (200 mL x 3). The combined organic layer was washed with brine (500 mL x 2), and dried over sodium sulfate. The organic layer was filtered and the solvent removed in vacuo to afford methyl 5-iodo-2- methoxy-3 -methylbenzoate (5 g, 13.88 mmol, 55.15% yield, 85% purity) as a yellow solid.
|0473] To methyl 5-iodo-2-methoxy-3-methylbenzoate (5 g, 16.33 mmol) and NBS (2.9 g, 16.33 mmol) in CCL (80 mL) was added AIBN (268 mg, 1.63 mmol), and the mixture was stirred at 80 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by column chromatography (SiO2,
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SUBSTITUTE SHEET ( RULE 26 ) petroleum ether/ethyl acetate = I/O to 20/1) to afford methyl 3-(bromomethyl)-5-iodo-2- methoxybenzoate (5.8 g, 11.00 mmol, 67.33% yield, 73% purity) as a yellow solid.
|0474] To a stirred solution of methyl 3-(bromomethyl)-5-iodo-2- methoxybenzoate (5.3 g, 13.77 mmol ) in 2-methoxyethanol (77.9 g, 1.02 mol, 80.82 mL), was added 2N NaOH (50 mL). The resulting mixture was stirred at 75 °C for 12 h. The residue was diluted with water (300 mL) and extracted with ethyl acetate (300 mL x 3). The clear solution was acidified with IN HC1 to pH ~ 5 and then extracted with ethyl acetate (200 mL x 3). The combined organic layer was washed with brine (500 mL), dried over Na SCL, filtered and concentrated under reduced pressure to afford 5-iodo-2-methoxy-3-((2 methoxyethoxy )methyl)benzoic acid (4.6 g, crude) as yellow oil.
[0475] To a solution of 6-(trifluoromethyl)-benzo[d]thiazol-2-amine (1.4 g, 6.55 mmol), 5-iodo-2-methoxy-3-((2-methoxyethoxy)methyl)benzoic acid (2 g, 5.46 mmol) in DCM (60 mL) was added EDCI (1.2 g, 6.55 mmol) and HOBt (1.1 g, 8.19 mmol). The mixture was stirred at 50 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (neutral condition) to afford 5-iodo-2-methoxy-3-((2-methoxyethoxy)methyl)-JV-(6- (trifluoromethylbenzo[<7]thiazol-2-yl]benzamide (920 mg, 1.62 mmol, 29.74% yield, 100% purity) as a white solid.
[0476] A mixture of 2,4,6-trimethyl-l,3,5,2,4,6-trioxatriborinane (199 mg, 794.59 pmol, 222.16 pL) 50% in THF, 5-iodo-2 -methoxy -3-((2-methoxyethoxy)methyl)-/V-(6- (trifluoromethylbenzo[</]thiazol-2-yl]benzamide (150 mg, 264.86 pmol), K2CO3 (73 mg, 529.73 pmol) and Pd(dppf)Cl2 CH2CI2 (21 mg, 26.49 pmol) in H2O (0.5 mL) and THF (2 mL), and the mixture was stirred at 80 °C for 12 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-TLC (SiCb, petroleum ether/ethyl acetate 0/1) to afford the mixture of 2- methoxy-3-((2 -methoxy ethoxy)methyl)-5-methyl-/V-(6-(trifluoromethyl)benzo[6(]thiazol-2- yl)benzamide and 2-m ethoxy-3 -((2 -methoxy ethoxy )methyl)-/V-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (50 mg) as a white solid.
[0477] To a mixture of EtSH (1 mL) and DCM (1 mL) was added Al Cl 3 (26 mg, 198.04 pmol, 10.82 pL) at 0 °C, the resulting solution was warmed to 20 °C, and then the mixture 2-m ethoxy-3 -((2 -methoxy ethoxy )methyl)-5-methyl-A-(6- (trifluoromethyl)benzo[c/]thiazol-2-yl)benzamide and 2-methoxy-3-((2- methoxyethoxy)methyl)-/V-(6-(trifluoromethyl)benzo[t ]thiazol-2-yl)benzamide (50 mg) was added with stirring, and the mixture was stirred at 20 °C for 9 h. The mixture was poured
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SUBSTITUTE SHEET ( RULE 26 ) into water (2 mL), HCI was added (2N) to reach a pH ~ 4-5, and the mixture was extracted with DCM (3 mL x 2). The combined organic layer was washed with brine (5 mL), dried over NazSCL, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (HCI condition) to afford 2-hydroxy-3-((2- methoxy ethoxy )methyl)-5-methyl-N-(6-(trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (5.15 mg, 11 69 pmol, 17.71% yield, 100% purity) as a black solid. LH NMR (400 MHz, METHANOL-^) 5 ppm 8.28 (s, 1 H), 7.84 (s, 2 H), 7.73 (d, J= 8.80 Hz, 1 H), 7.44 (s, 1 H), 4.65 (s, 2 H), 3.68 - 3.74 (m, 2 H), 3.59 - 3.64 (m, 2 H), 3.40 (s, 3 H), 2.35 (s, 3 H). MS (ESI) [M+H]+ requires m/z 441.10 , found m/z 441.1 and 2-m ethoxy-3 -((2- methoxy ethoxy )methyl)-5-methyl-A-(6-(trifluoromethyl)benzo[<7]thi azol -2 -yl)benzamide (6.62 mg, 15.22 pmol, 23.06% yield, 98.04% purity) as a white solid. LH NMR (400 MHz, METHANOL-^) 5 ppm 8.28 (s, 1 H), 8.00 - 8.06 (m, 1 H), 7.86 (br s, 1 H), 7.74 (d, J= 8.80 Hz, 1 H), 7.61 (br d, J= 7.34 Hz, 1 H), 7.02 (br t, J= 7.58 Hz, 1 H), 4.68 (s, 2 H), 3.69 - 3.75 (m, 2 H), 3.59 - 3.64 (m, 2 H), 3.40 (s, 3 H). MS (ESI) [M+H]+ requires m/z 427.09, found m/z 427.0.
Example 50
5-chloro-2-hydroxy-3-methoxy-7V-(5-(trifluoromethyl)thiazol-2-yl)benzamide (50)
10478] Sodium dithionite (5.2 g, 29.96 mmol, 6.52 mL) was added in one portion to a mixture of thiazol-2-amine (3 g, 29.96 mmol), NaHCCf (2.5 g, 29.96 mmol, 1.17 mL) and trifluoro(iodo)methane (23.4 g, 29.96 mmol) in ACN (40 mL) and H2O (10 mL) at 5 °C and stirred for 12 h under N2 atmosphere. The reaction mixture was concentrated under
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SUBSTITUTE SHEET ( RULE 26 ) reduced pressure to remove solvent. The residue was diluted with water 20 mL and extracted with ethyl acetate (10 mL x 3). The combined organic layer was washed with brine (50 mL), dried over NajSCL. filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCh, petroleum ether/ethyl acetate = 1/0 to 10/1) to afford 5-(trifluoromethyl)thiazol-2-amine (790 mg, 4.32 mmol, 14.43% yield, 92% purity) as yellow oil.
[0479] To a solution of 2-hydroxy-3 -methoxybenzoic acid (2 g, 11.89 mmol) in dioxane (20 mL), was added HC1 (6 M, 2.08 mL). H2O2 (1.4 g, 13.08 mmol, 1.26 mL, 30% purity) was added with rapid stirring of 1 min, and then the mixture was stirred at 20 °C for 12 h, producted from light. The reaction mixture was quenched by addition 10% Na2SC>3 (10 mL), concentrated under reduced pressure to remove solvent, then diluted with water (10 mL) and adjusted to pH 9 by addition of 2N NaOH. Then the mixture was extracted with ethyl acetate (15 mL x 3). The clear solution was acidified to pH 5 with IN HC1 and then extracted with ethyl acetate (15 mL x 3). The combined organic layer was washed with brine (50 mL), dried over Na2SC>4, filtered and concentrated under reduced pressure to give 5- chloro-2-hydroxy-3-methoxybenzoic acid (2.1 g, crude) as a yellow solid.
10480] To 5-chloro-2-hydroxy-3-methoxybenzoic acid (50 mg, 246.80 pmol) and 5 -(trifluoromethyl) thiazol-2-amine (41 mg, 246.80 pmol) in DCM (3 mL) was added EDCI (56 mg, 296.16 pmol) and HOBt (50 mg, 370.20 pmol), and the mixture was stirred at 45 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 5-chl oro-2 -hydroxy-3 - methoxy-JV-(5-(trifluoromethyl)thiazol-2-yl)benzamide (23.11 mg, 64.18 pmol, 26.00% yield, 97.95% purity) as a white solid. 'H NMR (400 MHz, MEI'HANOL- /4) 5 ppm 7.95 (d, J= 1.32 Hz, 1 H), 7.62 (d, J= 2.43 Hz, 1 H), 7.23 (d, J= 2.43 Hz, 1 H), 3.95 (s, 3 H). MS (ESI) [M+H]+ requires m/z 352.9 , found m/z 353.0
Example 51
5-chloro-2-hydroxy-3-(morpholinomethyl)-/V-(5-(trifluoromethyl)thiazol-2-yl)benzamide
(51)
SUBSTITUTE SHEET ( RULE 26 ) [0481| 5-chl oro-2 -hy droxy-3-(morpholinomethyl)-A-(5-(trifluoromethyl)thi azol-
2-yl)benzamide was made in a similar manner as Example 22. LCMS: MS (ESI) [M+H]+ requires m/z 422.0 , found m/z 422.0. XHNMR (400 MHz, METHANOL-d4) 8 ppm 8.14 (d, J = 2.45 Hz, 1 H), 8.00 (d, J = 1.47 Hz, 1 H), 7.59 (d, J = 2.45 Hz, 1 H), 4.35 (s, 2 H), 3.90 (br s, 4 H), 3.28 (br d, J = 4.40 Hz, 4 H)
Example 52
5-chloro-2-hydroxy-3-(pyrrolidin-l-ylmethyl)-A-(5-(trifluoromethyl)thiazol-2-yl)benzamide
(52)
[0482] 5-chl oro-2 -hy droxy-3 -(pyrrolidin-1 -ylmethyl)-jV-(5 -
(trifluoromethyl)thiazol-2-yl)benzamide was made in a similar manner to Example 22. LCMS: MS (ESI) [M+H]+ requires m/z 406.0 , found m/z 406.0. 1HNMR (400 MHz, METHANOL-d4) 8 ppm 8.25 (d, J= 2.65 Hz, 1 H), 8.07 (d, J= 1.10 Hz, 1 H), 7.74 (d, J = 2.43 Hz, 1 H), 4.46 (s, 2 H), 3.56 (br s, 2 H), 3.22 - 3.30 (m, 2 H), 2.21 (br s, 2 H), 2.05 (br s, 2 H)
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SUBSTITUTE SHEET ( RULE 26 ) Example 53
5-chloro-2-hydroxy-3-(morpholinomethyl)-7V-(6-(trifluoromethyl)pyridin-3-yl)benzamide
(53) and 5-chloro-2-hydroxy-3-(morpholinomethyl)-/V-(6-(trifluoromethyl)pyridin-3- yl)benzamide hydrochloride (53A)
[0483| To a stirred solution of 5-chloro-2-hydroxy-3-methylbenzoic acid (7 g, 37.51 mmol) in DMF (200 mL) was added K2CO3 (51.8 g, 375.15 mmol) followed by Mel (11.7 g, 82.53 mmol, 5.14 mL). The mixture was stirred at 20 °C for 12 h. H2O (500 mL) was added and extracted with ethyl acetate (300 mL x 3). The combined organic layer was washed with brine (500 mL) and dried over sodium sulfate. The organic layer was filtered and the solvent removed in vacuo. The residue was purified by column chromatography (SiC>2, petroleum ether/ethyl acetate = 1/0 to 10/1) to afford methyl 5-chloro-2-methoxy-3- methylbenzoate (87% purity) 11.2 g as a yellow solid.
[0484| Methyl 5-chloro-2-methoxy-3-methylbenzoate (2 g, 9.32 mmol) and 6- (trifluoromethyl)pyridin-3-amine (1.5 g, 9.32 mmol) in toluene (40 mL), was stirred at 0 °C, then AlMes (2 M, 13.98 mL) was added at 0 °C, the mixture was stirred at 20 °C for another 12 h. The reaction mixture was added to sat. NH4CI (100 mL), and then extracted with ethyl acetate (40 mL x 3). The combined organic layer was washed with brine (100 mL x 3), dried over Na2SC>4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCb, petroleum ether/Ethyl acetate = 1/0 to 20/1)
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SUBSTITUTE SHEET ( RULE 26 ) to afford 5-chloro-2-methoxy-3-methyl-/V-(6-(trifluoromethylpyridin-3-yl)benzamide (1.6 g, 2.51 mmol, 26.90% yield, 54% purity) as a yellow solid.
|0485] To 5-chloro-2-methoxy-3-methyl-Ar-(6-(trifluoromethylpyridin-3- yl)benzamide (1.5 g, 4.35 mmol) in CCk (20 mL) was added NBS (851 mg, 4.79 mmol) and AIBN (714 mg, 4.35 mmol), and the mixture was stirred at 70 °C for 12 (under light, 1000W). The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by column chromatography (Si O2, petroleum ether/ethyl acetate = 50/1 to 10/1) to afford 3-(bromomethyl)-5-chloro-2-methoxy-A-(6-(trifluoromethyl) pyridine- 3-yl)benzamide (1.4 g, 2.91 mmol, 66.84% yield, 88% purity) as a white solid.
[0486] To morpholine (113 mg, 1.30 mmol, 114.26 pL) in ACN (10 mL), was added K2CO3 (326 mg, 2.36 mmol). The mixture was stirred at 0 °C for 10 minutes, then 3- (bromomethyl)-5-chloro-2-methoxy-A-(6-(trifluoromethyl) pyridine-3-yl)benzamide (500 mg, 1.18 mmol) was added, and the mixture was stirred at 30 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by column chromatography (Si O?, petroleum ether/ethyl acetate = 1/0 to 1/1) to afford 5-chl oro-2 -methoxy-3-(morpholinomethyl)-A-(6-(trifluoromethyl)pyridin-3- yl)benzamide (340 mg, 711.93 pmol, 60.32% yield, 90% purity) as yellow oil.
[0487] AICI3 (139 mg, 1.05 mmol, 57.21 pL) was added to a mixture of EtSH (2 mL) and DCM (2 mL) at 0 °C, then stirred at 0 °C for 10 minutes. 5-Chloro-2-m ethoxy-3 - (morpholinomethyl)-A-(6-(trifluoromethyl)pyridin-3-yl)benzamide (150 mg, 348.98 pmol) was added, and the mixture was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep- HPLC (HC1 condition) to afford 5-chloro-2-hyoxy-3-(morpholinomethyl)-A-(6- (trifluoromethyl)pyridin-3-yl)benzamide (98.15% purity, HC1) 61.99 mg as a white solid. LH NMR (400 MHz, DMSO-<76) 6 ppm 9.06 (s, 1 H), 8.43 (dd, J= 8.80, 1.96 Hz, 1 H), 8.30 (br s, 1 H), 7.98 (d, J= 8.80 Hz, 1 H), 7.86 (br s, 1 H), 4.35 (br s, 2 H), 3.83 (br d, J= 9.29 Hz, 4 H), 3.16 - 3.32 (m, 2 H). MS (ESI) [M+H]+ requires m/z 416.0, found m/z 416.0.
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SUBSTITUTE SHEET ( RULE 26 ) Example 54
5-chloro-2-hydroxy-3-(piperidin-l-ylmethyl)-A-(6-(trifluoromethyl)pyridin-3-yl)benzamide
(54) and 5-chloro-2-hydroxy-3-(piperidin-l -ylmethyl)-A-(6-(trifluoromethyl)pyridin-3- yl)benzamide hydrochloride (54A)
(0488] To piperidine (30 mg, 354.10 pmol, 34.97 pL, Example 53) in ACN (2 mL) was added K2CO3 (97 mg, 708.20 pmol), and the mixture was stirred at 0 °C for 10 minutes. 3-(Bromomethyl)-5-chloro-2-methoxy-7V-(6-(trifluoromethyl)pyridine-3- yl)benzamide (150 mg, 354.10 pmol) was added, and the mixture was stirred at 30 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-TLC (SiCh, petroleum ether/ethyl acetate= 1 : 1) to afford 5- chloro-2-methoxy-3-(piperi din- l -ylmethyl )- -(6-(tri fluoromethyl )pyridin-3-yl (benzamide (100 mg, 186.98 pmol, 52.81% yield, 80% purity) as yellow oil.
[0489] 5-chl oro-2 -methoxy-3-(piperi din- 1 -ylmethyl)-A-(6-
(trifluoromethyl)pyridin-3-yl)benzamide (60 mg, 140.24 pmol) in DMF (2 mL) was stirred at 160 °C for 10 minutes, then NaSEt (58 mg, 701.19 pmol) was added, and the mixture was stirred at 160 °C for 1 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 5-chloro- 2 -hydroxy-3 -(piperi din- l-ylmethyl)-A-(6-(trifluoromethyl)pyri din-3 -yl)benzamide (18 mg, 39.32 pmol, 28.04% yield, 95.24% purity, HC1) as a white solid. 'H NMR (400 MHz, METHANOL-^) 8 ppm 9.07 (d, J= 2.21 Hz, 1 H), 8.47 (dd, J= 8.49, 2.32 Hz, 1 H), 8.30 (d, J= 2.43 Hz, 1 H), 7.88 (d, J= 8.60 Hz, 1 H), 7.79 (d, J= 2.43 Hz, 1 H), 4.38 (s, 2 H), 3.52 (br d, J= 12.35 Hz, 2 H), 3.10 (br t, J= 12.35 Hz, 2 H), 1.97 (br d, J= 14.55 Hz, 2 H), 1.70 - 1.87 (m, 3 H), 1.47 - 1.62 (m, 1 H). MS (ESI) [M+H]+ requires m/z 414.1, found m/z 414.1.
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SUBSTITUTE SHEET ( RULE 26 ) Example 55
5-chloro-3-((3-cyanopyrrolidin-l-yl)methyl)-2-hydroxy-A-(6-(trifluoromethyl)pyridin-3- yl)benzamide (55)
|0490] To 3-(bromomethyl)-5-chloro-2-methoxy-Ar-(6-(trifluoromethyl)pyridin-3- yl)benzamide (500 mg, 1.18 mmol, Example 53) in ACN (10 mL) was added K2CO3 (326 mg, 2.36 mmol), and the mixture was stirred at 0 °C for 10 minutes. Pyrrolidine-3 - carbonitrile (124 mg, 1.30 mmol, 7.88 pL) was added and the mixture was stirred at 30 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by column chromatography (SiCh, petroleum ether/Ethyl acetate = 1/0 to 1/ l) to afford 5 -chi oro-3 -((3 -cyanopyrrolidin-l-yl)methyl)-2 -methoxy -N-(6- (trifluoromethyl)pyridine-3-yl)benzamide (320 mg, 656.29 pmol, 55.62% yield, 90% purity) as yellow oil.
]049.1] To 5-chloro-3-((3-cyanopyrrolidin-l-yl)methyl)-2-methoxy-/V-(6- (trifluoromethyl)pyridine-3-yl)benzamide (150 mg, 341.82 pmol) in DCM (2 mL), was added BBrs (256 mg, 1.03 mmol, 98.81 pL) in DCM (0.5 mL) at -78 °C, and the mixture was stirred at 25 °C for 12 h. The reaction mixture was diluted with sat NaHCO, (5 mL) and extracted with DCM (5 mL x 3). The combined organic layer was washed with brine (5 mL), dried over NazSCh, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (HC1 condition) to afford 5-chloro-3-((3-cyanopyrrolidin-l- yl)methyl)-2-hydroxy-JV-(6-(trifluoromethyl)pyridin-3-yl)benzamide.
Example 56
5-chloro-2-hydroxy-/V-(6-(tri fluoromethyl )benzo[<7]thiazol-2-yl)-3-((2-
(trifhioromethyl)pyrrolidin-l-yl)methyl)benzamide (56) and 5-chloro-2-hydroxy-A-(6-
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SUBSTITUTE SHEET ( RULE 26 ) (trifluoromethyl)benzo[tZ]thiazol-2-yl)-3-((2-(trifluoromethyl)pyrrolidin-l- yl)methyl)benzamide hydrochloride (56A)
[04921 A solution of 5-chloro-2-hydroxybenzoic acid (50 g, 289.74 mmol), and 1,3,5,7-tetraazaadamantane (81.24 g, 579.49 mmol, 108.31 mL) in TFA (500 mL) was stirred at 100 °C for 12 h. The reaction mixture was cooled to 20 °C, followed by addition of HC1 (500 mL) (3M). After stirring for another 12 h, the solution was concentrated under reduced pressure. The white residue was suspended in water (1000 mL). The solid formed was collected, dried under vacuum to afford compound 5-chloro-3-formyl-2-hydroxybenzoic acid (50 g, crude) as a white solid.
(0493] A mixture of 5-chloro-3-formyl-2 -hydroxybenzoic acid (3 g, 14.96 mmol), 6-(trifluoromethyl)-benzo[t/]thiazol-2-amine (2.6 g, 11.97 mmol) and EDCI (3.4 g, 17.95 mmol), HOBt (3 g, 22.44 mmol) in DCM (150 mL) was heated at 50 °C for 8 h. The reaction mixture was concentrated to remove solvent. Then H2O (300 mL) was added and NaOH (3M) was added to reach a pH of ~ 9-10, extracted with ethyl acetate (200 mL x 2), then concentrated to remove ethyl acetate. After concentration 200 mL ethyl acetate was added and the precipitated solid was filtered and concentrated under reduced pressure to give 5- chloro-3-formyl-2-hydroxy-A-(6-(trifluoromethyl)benzo[tZ]thiazol-2-yl)benzamide (1.3 g) as a yellow solid.
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SUBSTITUTE SHEET ( RULE 26 ) [0494| To a solution of 2-(trifluoromethyl)pyrrolidine (38 mg, 274.48 pmol), 5- chloro-3-formyl-2-hydroxy-A-(6-(trifluoromethyl)benzo[</]thiazol-2-yl)benzamide (100 mg, 249 53 pmol) in MeOH (5 mL) was added acetic acid (14 mg, 249.53 pmol, 14.27 pL) to pH ~ 5-6. The mixture was stirred at 30 °C for 12 h then NaBHiCN (94 mg, 1.50 mmol) was added. The mixture was stirred at 30 °C for another 4 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep- HPLC (HC1 condition) to afford 5-chloro-2-hydroxy-A-(6-(trifluoromethyl)benzo[d]thiazol- 2-yl)-3-((2-(trifluoromethyl)pyrrolidin-l-yl)methyl)benzamide (18 mg, 32.16 pmol, 12.89% yield, 100% purity, HC1) as a white solid. XH NMR (400 MHz, METHANOL-c/4) 8 ppm 8.30 (s, 1 H), 8.04 (d, J= 2.45 Hz, 1 H), 7.87 (d, J= 8.80 Hz, 1 H), 7.75 (d, J= 8.31 Hz, 1 H), 7.49 (d, J= 2.45 Hz, 1 H), 4.41 (br d, J= 14.67 Hz, 1 H), 4.04 (d, J = 14.18 Hz, 1 H), 3.73 (br s, 1 H), 3.11 - 3.25 (m, 1 H), 2.65 - 2.78 (m, 1 H), 2.21 - 2.33 (m, 1 H), 2.08 (dt, J = 17.24, 3.36 Hz, 1 H), 1.97 (br d, J= 9.29 Hz, 2 H). MS (ESI) [M+H]+ requires m/z 524.0, found m/z 524.0.
Example 57
5-chloro-2-hydroxy-A-('6-(tri fluoromethyl )benzo[<7]thiazol-2-yl)-3-(('4- (trifluoromethyl)piperidin-l-yl)methyl)benzamide (57)
[0495] 5-chl oro-2 -hydroxy -A'-(6-(tri fluoromethyl )benzo[t/]thi azol -2 -yl)-3-((4-
(trifluoromethyl)piperidin-l-yl)methyl)benzamide was made in a manner similar to Example 56. LCMS: MS (ESI) [M+H]+ requires m/z 538.0 found m/z 538.0. 1HNMR (400 MHz, METHANOL-^) 8 ppm 8.21 - 8.34 (m, 2 H), 7.75 - 7.86 (m, 2 H), 7.67 (d, J= 2.45 Hz, 1 H), 4.41 (s, 2 H), 3.67 (br d, J= 10.76 Hz, 2 H), 3.09 - 3.26 (m, 2 H), 2.63 (br s, 1 H), 2.18 (br d, J= 14.67 Hz, 2 H), 1.88 (br d, J= 12.72 Hz, 2 H)
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SUBSTITUTE SHEET ( RULE 26 ) Example 58
5-chloro-3-((3-fluoropiperidin- 1 -yl)methyl)-2 -hydroxy -N-(6- (trifluoromethyl)benzo[t/]thiazol-2-yl)benzamide
(0496| 5-chl oro-3-((3-fluoropiperidin-l-yl)methyl)-2 -hydroxy -N-(6-
(trifluoromethyl)benzo[c/]thiazol-2-yl)benzamide was made in a similar manner to Example 56. LCMS: MS (ESI) [M+H]+ requires m/z 488.07, found m/z 488.1. 1HNMR (400 MHz, METHANOL-^) 8 ppm 8.27 (s, 1 H), 8.21 (d, J= 2.65 Hz, 1 H), 7.78 (s, 2 H), 7.63 (d, J = 2.43 Hz, 1 H), 4.97 - 5.15 (m, 1 H), 4.32 - 4.46 (m, 2 H), 3.66 (br s, 1 H), 3.37 - 3.56 (m, 2 H), 3.10 - 3.21 (m, 1 H), 2.12 (br s, 2 H) 1.87 (br d, J= 8.82 Hz, 2 H)
Example 59
5-chloro-3-((3,3-difluoropiperidin-l-yl)methyl)-2-hydroxy-/V-(6- (tri fl uoromethy I )benzo[c/]thi azol -2-yl (benzamide (59)
|0497( 5-chloro-3-((3,3-difluoropiperidin-l-yl)methyl)-2-hydroxy-A-(6- (trifluoromethyl)benzo[c/]thiazol-2-yl (benzamide was made in a manner similar to Example 56. LCMS: MS (ESI) [M+H]+ requires m/z 506.0 , found m/z 506.0. LHNMR (400 MHz, DMSO-4) 8 ppm 8.50 (s, 1 H) 7.85 - 7.92 (m, 2 H) 7.77 (br d, J=8.80 Hz, 1 H) 7.48 (d, .7=2,45 Hz, 1 H) 4.10 (br s, 2 H) 3.29 (br s, 2 H) 2.87 (br s, 2 H) 2.05 (br s, 2 H) 1.83 (br s, 2 H)
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SUBSTITUTE SHEET ( RULE 26 ) Example 60
5-chloro-3-((4-fluoropiperidin- 1 -yl)methyl)-2 -hydroxy -N-(6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (60)
[0498] 5-chl oro-3-((4-fluoropiperidin-l-yl)methyl)-2 -hydroxy -N-(6-
(trifluoromethyl)benzo[d]thiazol-2-yl)benzamide was made in a manner similar to Example 56. LCMS: MS (ESI) [M+H]+ requires m/z 488.0, found m/z 488.1. 1HNMR (400 MHz, METHANOL-^) 8 ppm 8.29 (s, 1 H) 8.25 (d, J=2.65 Hz, 1 H) 7.79 (s, 2 H) 7.67 (s, 1 H) 5.02 (br s, 1 H) 4.42 (s, 2 H) 3.35 - 3.51 (m, 4 H) 2.19 (br s, 4 H)
Example 61
5-chloro-2-hydroxy-/V-(6-(trifluoromethyl)benzo[t/]thiazol-2-yl)-3-((3-
(trifluoromethyl)piperidin-l-yl)methyl)benzamide (61)
[0499] 5 -chi oro-2 -hydroxy -jV-(6-(trifluoromethyl)benzo[d]thi azol -2 -yl)-3-((3-
(trifluoromethyl)piperidin-l-yl)methyl)benzamide was made in a manner similar to Example 56. LCMS: MS (ESI) [M+H]+ requires m/z 538.0 , found m/z 538.1. 1HNMR (400 MHz, METHANOL-d4) 8 ppm 8.27 (br s, 1 H) 8.22 (br d, J=4.19 Hz, 1 H) 7.78 (br s, 2 H) 7.65 (d, J=2.65 Hz, 1 H) 4.37 - 4.48 (m, 2 H) 3.69 (br d, J=11.25 Hz, 1 H) 3.52 (br d, J=12.35 Hz, 1 H) 3.17 (t, J=12.24 Hz, 1 H) 3.02 - 3.11 (m, 1 H) 2.85 (br s, 1 H) 2.03 - 2.15 (m, 2 H) 1.85 (br d, J=12.79 Hz, 1 H) 1.57 - 1.70 (m, 1 H)
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SUBSTITUTE SHEET ( RULE 26 ) Example 62
5-chloro-3-((4,4-difluoropiperidin-l-yl)methyl)-2-hydroxy-?/-(6- (trifluoromethy l)b cnzo[t/] th i azol -2-y l)b enzami de (62)
[0500] 5-chloro-3-((4,4-difluoropiperidin-l-yl)methyl)-2-hydroxy-7V-(6- (trifluoromethyl)benzo[c/]thiazol-2-yl)benzamide was made in a manner similar to Example 56. LCMS: MS (ESI) [M+H]+ requires m/z 506.0 , found m/z 506.1. LHNMR (400 MHz, METHANOL-^) 8 ppm 8.29 (s, 1 H) 8.27 (d, J=2.87 Hz, 1 H) 7.77 - 7.84 (m, 2 H) 7.69 (d, J=2.65 Hz, 1 H) 4.47 (s, 2 H) 3.47 - 3.57 (m, 4 H) 2.30 - 2.46 (m, 4 H)
Example 63
5-chl oro-3 -ethyl-2-hydroxy-A-(4-((2 -methoxy ethoxy)methyl)-6- (trifluoromethy l)b enzo[c/] th i azol -2-y l)b enzami de (63 )
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SUBSTITUTE SHEET ( RULE 26 )
[0501 To a solution of methyl 5-chloro-2-methoxy-3-vinylbenzoate (0.45 g, 1.99 mmol) in MeOH (6 mL) was added Pd/C (0.1 g, 10% purity). The mixture was degassed and purged with H2 3 times, and then the mixture was stirred at 20 °C for 30 minutes under H2 (15 Psi) atmosphere. The reaction mixture was filtered and concentrated under reduced pressure to afford methyl 5-chl oro-3 -ethyl-2 -methoxybenzoate (400 mg) as a yellow oil. XH NMR (400 MHz, CHLOROFORM-tZ) 5 ppm 7.61 - 7.65 (m, 1 H), 7.34 (d, J= 2.65 Hz, 1 H), 3.90 - 3.95 (s, 3 H), 3.81 - 3.85 (s, 3 H), 2.63 - 2.74 (m, 2 H), 1.19 - 1.28 (m, 3 H)
[0502] To a solution of methyl 5-chl oro-3 -ethyl-2 -methoxybenzoate (0.3 g, 1.31 mmol) in DCM (6 mL) was added dropwise RBn (1.97 g, 7.87 mmol, 758.45 pL) at -78 °C, then the resulting mixture was stirred at 40 °C for 12 h. The reaction mixture was quenched by addition sat. NH4CI (10 mL), and then extracted with DCM (3 mL x 2). The combined organic layer was washed with brine (5 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to afford methyl 5-chloro-3-ethyl-2-hydroxybenzoate (0.25 g) as a yellow solid.
[0503] To a solution of methyl 5-chl oro-3 -ethyl-2 -hydroxybenzoate (0.25 g, 1.16 mmol) in THF (3 mL) and H2O (1 mL) was added LiOH H2O (146 mg, 3.49 mmol). The mixture was stirred at 25 °C for 12 h. The reaction mixture was extracted with ethyl acetate (5 mL x 3), then the aqueous solution was acidized to pH to 5-6 with HC1 (IM), and extracted with ethyl acetate (5 mL x 3). The combined organic layer was washed with brine (30 mL), dried over Na2SC>4, filtered and concentrated under reduced pressure to afford 5- chl oro-3 -ethyl-2-hydroxybenzoic acid (200 mg) as a yellow solid.
[0504] To a solution of 5-chloro-3-ethyl-2-hydroxybenzoic acid (80 mg, 398.77 pmol) and 4-((2 -methoxy ethoxy )methyl)-6-(trifluoromethyl)benzo[t/]thiazol-2-amine (122
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SUBSTITUTE SHEET ( RULE 26 ) mg, 398.77 pmol, Example 13) in DCM (3 mL) was added EDCI (91 mg, 478.52 pmol) and HOBt (80 mg, 598.15 pmol). The mixture was stirred at 45 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HCI condition) to afford 5-chl oro-3 -ethyl-2 -hydroxy - V-(4-((2- methoxyethoxy)methyl)-6-(trifluoromethyl)benzo[<7]thiazol-2-yl)benzamide (108.9 mg, 222.74 pmol, 55.86% yield, 100% purity) as a white solid. 'H NMR (400 MHz, DMSO-flk) 6 ppm 8.45 (s, 1 H), 8.05 (br s, 1 H), 7.80 (s, 1 H), 7.44 (d, J= 1.96 Hz, 1 H), 4.96 (s, 2 H), 3.70 (dd, J= 5.62, 3.67 Hz, 2 H), 3.48 - 3.60 (m, 2 H), 3.28 (s, 3 H), 2.65 (q, J= 7.83 Hz, 2 H), 1.18 (t, J = 7.58 Hz, 3 H). MS (ESI) [M+H]+ requires m/z 489 0, found m/z 489.0
Example 64
5-chl oro-3-((cA-2,6-dimethylmorpholino)methyl)-2 -hydroxy -A-(4-((2- methoxyethoxy)methyl)-6-(trifluoromethyl)benzo[t ]thiazol-2-yl)benzamide (64) and 5- chloro-3-((czs-2,6-dimethylmorpholino)methyl)-2-hydroxy-A-(4-((2-methoxyethoxy)methyl)-
6-(trifluoromethyl)benzo[z7]thiazol-2-yl)benzamide hydrochloride (64A)
|0505] To a solution of 5-chloro-2-hydroxybenzoic acid (50 g, 289.74 mmol) in TFA (500 mL) was added 1,3,5,7-tetraazaadamantane (81.2 g, 579.49 mmol) at 25 °C. The mixture was stirred at 100 °C for 12 h, then cooled to 20 °C, followed by addition of HCI (500 mL, 3 M). After stirring for another 12 h, the solution was concentrated under reduced pressure. The white residue was suspended in water (1000 mL). The solid was collected, dried under vacuum to afford compound 5-chloro-3-formyl-2- hydroxybenzoic acid (50 g, crude) as a white solid.
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SUBSTITUTE SHEET ( RULE 26 ) [0506] To a solution of 5-chloro-3-formyl-2-hydroxybenzoic acid (2 g, 9.97 mmol), and cA-2,6-dimethylmorpholine (2.3 g, 19.94 mmol, 2.46 mL) in toluene (50 mL) was added acetic acid (598 mg, 9.97 mmol, 570.26 pL) to make pH ~ 5-6. The mixture was stirred at 80 °C for 12 h, then the reaction mixture was concentrated under reduced pressure to remove solvent and then MeOH (50 mL) and NaBHtCN (3.7 g, 59.83 mmol) were added. The mixture was stirred at 50 °C for another 3 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (neutral condition) to afford compound 5-chloro-3-((cA-dimethylmorpholino)methyl]-2- hydroxybenzoic acid (1.73 g, 4.62 mmol, 46.30% yield, 80% purity) as a yellow solid. LH NMR (400 MHz, DMSO-<76) 8 ppm 7.67 (d, J= 2.93 Hz, 1 H), 7.41 (d, J= 2.93 Hz, 1 H), 3.78 - 3.92 (m, 2 H), 3.21 - 3.33 (m, 2 H), 2.56 - 2.68 (m, 2 H), 1.11 (d, J= 6.36 Hz, 6 H).
[0507] A mixture of 2-methyl-4-(trifluoromethyl)aniline (22 g, 125.61 mmol), ammonia thiocyanic acid (14.3 g, 188.41 mmol, 14.34 mL) and TFA (35.8 g, 314.02 mmol, 23.25 mL) in CHCL (440 mL) was stirred at 80 °C for 12 h. To the reaction mixture was
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SUBSTITUTE SHEET ( RULE 26 ) added water (1000 mL) and extracted with ethyl acetate (500 mL x 3). The combined organic layer was washed with brine (600 mL x 3), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 5-50% ethyl acetate/petroleum ether gradient @ 100 mL/min) to afford l-(2-methyl-4- (trifluoromethyl)phenyl)thiourea (172 g, 69.02 mmol, 54.95% yield, 94% purity) as a yellow solid. LH NMR (400 MHz, CHLOROFORM-d) 5 ppm 8.09 (br s, 1 H), 7.50 - 7.65 (m, 2 H), 7.39 (br d, J = 7.72 Hz, 1 H), 6.12 (br s, 2 H), 2.40 (br s, 3 H). MS (ESI) [M+H]+ requires m/z 235.0, found m/z 235.1
[05081 At 0 °C, a solution of Br2 (9.8 g, 61.48 mmol, 3.17 mL) in CHCL (15 mL) was added dropwise to a stirred solution of (2-methyl-4-(trifluoromethyl)phenyl)thiourea (12 g, 51.23 mmol) in CHCh (100 mL), then the mixture was stirred at 70 °C for 12 h, then cooled to 20 °C. The precipitated white solid was filtered out and collected, then the solid was diluted with water (500 mL) and neutralized by NH3.H2O to pH ~ 8-9, and extracted with ethyl acetate (250 mL x 3). The combined organic layer was washed with brine (500 mL), dried over Na2SC>4. filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCh, petroleum ether/ethyl acetate = 1/0 to 5/1) to afford 4-methyl-6-(trifluoromethyl)benzo[t/]thiazol-2-amine (91% purity) 13.4 g as a yellow solid. 'H NMR (400 MHz, CHLOROFORM-^/) 8 ppm 7.70 (s, 1 H), 7.37 (s, 1 H), 6.30 (br s, 2 H), 2.56 (s, 3 H)
[0509] To 4-methyl-6-(trifluoromethyl)benzo[</]thiazol-2-amine (13.4 g, 57.70 mmol) and BOC2O (31.4 g, 144.26 mmol, 33.14 mL) in DCM (400 mL) was added DMAP (704 mg, 5.77 mmol), and the mixture was stirred at 20 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (ISCO®;40 g SepaFlash® Silica Flash Column, Eluent of 0-2% ethyl acetate/petroleum ether gradient @ 70 mL/min) to afford tert-butyl N-tert- butoxycarbonyl-A-(4-methyl-6-(trifluoromethyl)benzo[ ]thiazol-2-yl)carbamate (24.76 g, 48.67 mmol, 84.34% yield, 85% purity) as yellow oil. XHNMR (400 MHz, CHLOROFORM-tZ) 8 ppm 7.90 (s, 1 H), 7.46 (s, 1 H), 2.64 (s, 3 H), 1.62 (s, 18 H)
[0510] To tert-butyl A-tert-butoxycarbonyl-/V-(4-methyl-6- (trifluoromethyl)benzo[d]thiazol-2-yl)carbamate (20 g, 46.25 mmol) and NBS (24.6 g, 138.74 mmol) in CCI4 (500 mL) was added AIBN (759 mg, 4.62 mmol), and the mixture was stirred at 80 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by column chromatography (SiCh, petroleum
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SUBSTITUTE SHEET ( RULE 26 ) ether/ethyl acetate = 1/0 to 20/1) to afford Zc/7-butyl /V-Zc77-butoxycarbonyl-A-(4- (dibromomethyl)-6-(trifluoromethyl)benzo[z/]thiazol-2-yl)carbamate (27 g, 36.32 mmol, 78.53% yield, 79.392% purity) as yellow oil. 'H NMR (400 MHz, CHLOROFORM-^/) 8 ppm 8.15 (s, 1 H), 8.00 - 8.05 (m, 1 H), 7.42 (s, 1 H), 4.13 (q, J= 7.34 Hz, 1 H), 1.60 - 1.69 (s, 18 H)
[05111 Diethyl phosphate (6 3 g, 45.74 mmol, 5.90 mL) was added to a stirred mixture of tert-butyl JV-(4-(dibromomethyl)-6-(trifluoromethyl)benzo[</]thiazol-2-yl)-jV-tert- butoxycarbonyl carbamate (27 g, 45.74 mmol) in isopropyl acetate (200 mL), N,N- diisopropylethylamine (3.2 g, 25 16 mmol, 4.38 mL) was added dropwise and the mixture was stirred at 20 °C for 12 h. The reaction mixture was diluted with water (500 mL) and extracted with ethyl acetate (200 mL x 2). The combined organic layer was washed with brine (500 mL), dried over NaiSCh, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCb, Petroleum ether / Ethyl acetate 1/0 to 20/1) to afford t di-ZcvZ-butyl (4-(bromomethyl)-6- (trifluoromethyl)benzo[t/]thiazol-2-yl)iminodicarbonate as a yellow solid .
|0512] To a stirred solution of the di-Zc77-butyl (4-(bromomethyl)-6- (trifluorornethyl)benzo[d]thiazol-2-yl)iminodicarbonate (5 g, 7.33 mmol) in 2- methoxyethanol (144.7 g, 1.90 mol, 149.95 mL) was added NaOH (1 M, 56.25 mL). The resulting mixture was stirred at 20 °C for 12 h. The reaction mixture was diluted with water (150 mL) and extracted with ethyl acetate (50 mL x 2). The combined organic layer was washed with brine (500 mL x 2), dried over ISfeSCL, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0-10% ethyl acetate/petroleum ether gradient @ 70 mL/min) to afford ZcvZ-butyl (4-((2-methoxyethoxy)methyl)-6- (trifluoromethyl)benzo[z/]thiazol-2-yl]carbamate (2.6 g, 5.30 mmol, 72.30% yield, 82.88% purity) as a white solid.
[0513] To a solution of Zc77-butyl (4-((2-methoxyethoxy)methyl)-6- (trifluoromethyl)benzo[J]thiazol-2-yl]carbamate (0.6 g, 1.48 mmol) in DCM (3 mL) was added TFA (3 mL). The mixture was stirred at 20 °C for 0.5 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The reaction mixture was diluted with water (5 mL) and extracted with DCM (3 mL x 3). The combined organic layers were washed with brine (20 mL), dried over NazSCL, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, eluent of 0-50% ethyl acetate/petroleum
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SUBSTITUTE SHEET ( RULE 26 ) ether gradient @70 mL/min) to afford 4-((2-methoxyethoxy)methyl)-6- (trifluoromethyl)benzo[J]thiazol-2-amine (94.47% purity 1.2 g) as a yellow solid. 'H NMR (400 MHz, DMSO-dfi) 5 ppm 8.04 (s, 1 H), 7.98 (br s, 2 H),7.53 (s, 1 H), 4.76 (s, 2 H), 3.64 (dd, J= 5.62, 3.67 Hz, 2 H), 3.51 (dd, J= 5.62, 4.16 Hz, 2 H), 3.24 - 3.29 (m, 3 H)
[05l4j To a solution of 4-((2 -methoxyethoxy )methyl)-6- (trifluoromethyl)benzo[r/]thiazol-2-amine (0.5 g, 1.63 mmol), 5-chloro-3-((czs-2,6- dimethylmorpholino)methyl)-2-hydroxybenzoic acid (733 mg, 2.45 mmol) in DCM (30 m ) was added EDCI (375 mg, 1.96 mmol) and HOBt (330 mg, 2.45 mmol). The mixture was stirred at 45 °C for 12 h The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 5-chloro- 3-((cA-2,6-dimethylmorpholino)methyl)-2-hydroxy-/V-(4-((2-methoxy ethoxy )methyl)-6- (trifluoromethyl)benzo|@]thiazol-2-yl]benzamide (544.77 mg, 836.59 pmol, 51.25% yield, 95.90% purity, HC1) as a yellow solid. LH NMR (400 MHz, METHANOL-^) 8 ppm 8.24 (d, J= 2.45 Hz, 1 H), 8.20 (s, 1 H), 7.79 (s, 1 H), 7.71 (d, J = 2.45 Hz, 1 H), 4.98 (s, 2 H), 4.41 (s, 2 H), 3.91 (br s, 2 H), 3.80 (dd, J= 5.62, 3.67 Hz, 2 H), 3.65 - 3.71 (m, 2 H), 3.41 - 3.48 (m, 5 H), 2.84 (t, J = 11.74 Hz, 2 H), 1.25 (d, J = 6.36 Hz, 6 H). MS (ESI) [M+H]+ requires m/z 588.1, found m/z 588.1
Example 65
5-chloro-2-hydroxy-A-(4-((2-methoxy ethoxy )methyl)-6-(trifluoromethyl)benzo[<7] thiazol-2- yl)-3-(morpholinomethyl)benzamide (65)
|0515] 5-chloro-2-hydroxy-A-(4-((2-methoxyethoxy)methyl)-6- (trifluoromethyl)benzo[r ]thiazol-2-yl)-3-(morpholinomethyl)benzamide was made in a manner similar to Example 64. LCMS: MS (ESI) [M+H]+ requires m/z 560.1, found m/z 560.2. 1HNMR (400 MHz, METHANOL-d4) 8 ppm 8.12 - 8.19 (m, 2 H), 7.74 (s, 1 H), 7.68
-231-
SUBSTITUTE SHEET ( RULE 26 ) (d, J = 1.96 Hz, 1 H), 4.95 (s, 2 H), 4.40 (s, 2 H), 3.93 (br s, 4 H), 3.80 (dd, J = 5.38, 3.42 Hz,
2 H), 3.64 - 3.72 (m, 2 H), 3.44 (s, 3 H), 3.37 (br s, 4 H)
Example 66
5-chloro-2-hydroxy-jV-(4-((2-methoxy ethoxy )methyl)-6-(trifluoromethyl)benzo[d]thiazol-2- yl)-3-(piperidin- 1 -ylmethyl)benzamide (66)
|0516] 5-chl oro-2 -hydroxy -7V-(4-((2-methoxy ethoxy )methyl)-6-
(trifluoromethyl)benzo[d]thiazol-2-yl)-3-(piperidin-l-ylmethyl)benzamide was made in a manner similar to Example 64. LCMS: MS (ESI) [M+H]+ requires m/z 558.1, found m/z 558.2. 1HNMR (400 MHz, DMSO-d6) 5 ppm 8.40 (s, 1 H), 8.01 (br s, 1 H), 7.75 (s, 1 H) 7.69 (br s, 1 H), 4.94 (s, 2 H), 4.25 (s, 2 H), 3.69 (dd, J = 5.14, 3.67 Hz, 2 H), 3.42 - 3.58 (m, 6 H), 3.27 (s, 3 H), 1.37 - 1.82 (m, 6 H)
Example 67
5-chloro-2-hydroxy-2V-(4-methoxy-6-(trifluoromethyl)benzo[</]thiazol-2-yl)-3- methylbenzamide (67)
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SUBSTITUTE SHEET ( RULE 26 ) [0517] To a solution of 2-amino-5-(trifluoromethyl)phenol (0.5 g, 2.82 mmol) in MeOH (396 mg, 12.36 mmol, 500.34 pL) and THF (10 mL) was added PPh3 (1.4 g, 5.65 mmol), the mixture was stirred at 20 °C for 5 minutes, then DIAB (1.5 g, 7.62 mmol, 1.48 mL) was added dropwise, and the mixture was stirred at 20 °C for another 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by column chromatography (SiCb, petroleum ether/ethyl acetate = 1/0 to 20/1) to afford 2-methoxy-4-(trifluoromethyl)aniline (380 mg, 1.77 mmol, 62.68% yield, 89% purity) as yellow oil.
(0518] A mixture of 2-methoxy-4-(trifluoromethyl)aniline (380 mg, 1.99 mmol), ammonium thiocyanate (226 mg, 2.98 mmol, 226.99 pL) and TFA (566 mg, 4.97 mmol, 367.97 pL) in CHCh (8 mL) was stirred at 80 °C for 12 h. To the reaction mixture was added water (5 mL) and extracted with DCM (5 mL x 3). The combined organic layer was washed with brine (10 mL x 2), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether/ethyl acetate = 1/0 to 5/1) to afford l-(2-methoxy-4- (trifluoromethyl)phenyl)thiourea (220 mg, 733.24 pmol, 36.88% yield, 83.40% purity) as a yellow solid. MS (ESI) [M+H]+ requires m/z 251.0, found m/z 251.2.
(0519] At 0 °C, a solution of Br2 (119 mg, 747.28 pmol, 38.52 pL) in CHCh (1.5 mL) was added dropwise to a stirred solution of 1 -(2 -methoxy -4- (trifluoromethyl)phenyl)thiourea (170 mg, 679.35 pmol) in CHCh (6 mL), then the mixture was stirred at 70 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H2O (5 mL) and neutralized by NH3.H2O to pH ~ 8-9, and extracted with ethyl acetate (5 mL x 3). The combined organic layer was washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO2, petroleum ether/ethyl acetate = 1:1) to afford 4-methoxy-6-(trifluoromethyl)benzo[d]thiazol-2-amine (62 mg, 238.07 pmol, 35.04% yield, 95.31% purity) as a yellow solid. 'H NMR (400 MHz, CHLOROFORM-rZ) 5 ppm 7.51 (s, 1 H), 7.02 (s, 1 H), 5.62 (br s, 2 H), 4.02 (s, 3 H). MS (ESI) [M+H]+ requires m/z 249.0 , found m/z 249.0.
]0520] To a solution of 4-methoxy-6-(trifluoromethyl)benzo[rZ]thiazol-2-amine (60 mg, 241.72 pmol) and 5-chloro-2-hydroxy-3-methylbenzoic acid (67 mg, 362.57 pmol) in DCM (5 mL) was added EDCI (55 mg, 290.06 pmol) and HOBt (48 mg, 362.57 pmol). The mixture was stirred at 45 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (neutral
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SUBSTITUTE SHEET ( RULE 26 ) condition) to afford 5-chloro-2-hydroxy-7V-(4-methoxy-6-(trifluoromethyl)benzo|/7]thiazol-2- yl)-3 -methylbenzamide (20 mg, 49.33 pmol, 20.41% yield, 98.84% purity) as a white solid. 'H NMR (400 MHz, MF.THANOI.-J4) 8 ppm 7 88 - 7.90 (m, 1 H), 7.81 (s, 1 H), 7 30 (d, J = 1.96 Hz, 1 H), 7.23 (s, 1 H), 4.07 (s, 3 H), 2.24 (s, 3 H). MS (ESI) [M+H]+ requires m/z 417.0 , found m/z 417.0.
Example 68
5-chloro-2-hydroxy-.¥-(4-(methoxyniethyl)-6-(trifluoromethyl )benzo[<7]tliiazol-2-yl)-3- methylbenzamide (68)
(trifluoromethyl)benzo[d]thiazol-2- yl)iminodicarbonate
[0521] di-/c77-biityl (4-(bromomethyl)-6-(trifluoromethyl)benzo[z/]thiazol-2- yl)iminodicarbonate (0.5 g, 1.22 mmol, Example 64) was added to NaOMe (10 mL) (30%). The mixture was stirred at 30 °C for 12 h. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (5 mL x 2). The combined organic layer was washed with brine (30 mL), dried over NazSCL, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® Silica Flash Column, Eluent of 0~7% ethyl acetate/petroleum ether gradient @ 70 mL/min) to afford tert-butyl (tert-butoxycarbonyl)(4-(methoxymethyl)-6- (trifluoromethyl)benzo[J]thiazol-2-yl)carbamate (280 mg, 688.37 pmol, 56.62% yield, 89.08% purity) as a white solid.
-234-
SUBSTITUTE SHEET ( RULE 26 ) [0522 | To a solution of tert-butyl A-[4-(methoxymethyl)-6-(trifluoromethyl)-l,3- benzothiazol-2-yl]carbamate (280 mg, 772.70 pmol) in DCM (3 mL) was added TFA (1 mL). The mixture was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with water (3 mL) and extracted with ethyl acetate (3 mL x 2). The combined organic layer was washed with brine (4 mL x 2), dried over NaiSCL, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 50% ethyl acetate/petroleum ether gradient @ 70 mL/min) to afford 4-(methoxymethyl)-6-(trifluoromethyl)benzo[<7]thiazol-2-amine (180 mg, 558 74 pmol, 72.31% yield, 81.40% purity) as a yellow solid.
[0523] To a solution of 4-(mcthoxymcthyl)-6-('trifluoromcthyl)bcnzo[</]thiazol-2- amine (130 mg, 495.71 pmol), and 5-chloro-2-hydroxy-3-methylbenzoic acid (138 mg, 743.56 pmol) in DCM (6 mL), was added EDCI (114 mg, 594.85 pmol) and HOBt (100 mg, 743.56 pmol). The mixture was stirred at 45 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep- HPLC (neutral condition) to afford 5-chloro-2-hydroxy-/V-(4-(methoxymethyl)-6- (trifluoromethyl)benzo[d]thiazol -2 -yl)-3 -methylbenzamide (63.3 mg, 139.82 pmol, 28.21% yield, 95.16% purity) as a yellow solid. TH NMR (400 MHz, DMSO-cZs) 5 ppm 8.43 (br s, 1 H), 8.00 (br s, 1 H) 7.72 (br s, 1 H), 7.44 (br s, 1 H), 4.85 (br s, 2 H), 3.41 (br s, 3 H), 2.21 (br s, 3 H). MS (ESI) [M+H]+ requires m/z 431.0, found m/z 431.1.
Example 69
3-((bis(2 -methoxy ethyl)amino)methyl)-5-chl oro-2 -hydroxy -N-(4-(methoxymethyl)-6- (trifluoromethy l)b enzo[<7] th i azol -2-y l)b enzami de (69)
[0524] 3-((bis(2-methoxyethyl)amino)methyl)-5-chl oro-2 -hydroxy-N-(4-
(methoxymethyl)-6-(trifluoromethyl)benzo[<7]thiazol-2-yl (benzamide was made in a manner similar to Example 68. LCMS: MS (ESI) [M+H]+ requires m/z 562.1, found m/z 562.2. 1HNMR (400 MHz, METHANOL-d4) 5 ppm 7.99 (s, 2 H), 7.55 (s, 1 H), 7.44 (d, J = 2.21
-235-
SUBSTITUTE SHEET ( RULE 26 ) Hz, 1 H), 4.68 (s, 2 H), 4.35 (s, 2 H), 3.55 - 3.62 (m, 4 H), 3.28 - 3.31 (m, 5 H), 3.23 (s, 6 H), 3.12 (dt, J = 3.14, 1.63 Hz, 2 H).
Example 70
3-((bis(2-methoxyethyl)amino)methyl)-5-chloro-2-hydroxy-7V-(4-((2- methoxy ethoxy )methyl)-6-(trifluoromethyl)benzo[</]thiazol-2-yl)benzamide (70)
[0525] To a solution of 5-chloro-3-formyl-2-hydroxybenzoic acid (0.5 g, 2.49 mmol) and 2-methoxy- V-(2-methoxyethyl)ethanamine (664 mg, 4.99 mmol, 736.17 pL) in toluene (10 mL) was added acetic acid (149 mg, 2.49 mmol, 142.57 pL ) to pH ~ 5-6. The mixture was stirred at 80 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent and then MeOH (10 mL) and NaBHsCN (939 mg, 14.96 mmol) were added. The mixture was stirred at 50 °C for another 3h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep- HPLC (neutral conditions) to afford 3-((bis(2-methoxyethyl)amino)methyl)-5-chloro-2- hydroxybenzoic acid (400 mg, 352.46 pmol, 14.14% yield, 28% purity) as a white solid. LH NMR (400 MHz, DMSO-o ) 5 ppm 7.63 (d, J= 2.93 Hz, 1 H), 7.38 (d, J= 2.93 Hz, 1 H), 4.34 (s, 2 H), 3.67 (t, J= 4.89 Hz, 4 H), 3.31 (br t, J = 4.65 Hz, 4 H), 3.27 (s, 6 H).
[0526] To a solution of 3-((bis(2 -methoxy ethyl)amino)methyl)-5-chloro-2- hydroxybenzoic acid (155 mg, 489.71 pmol), 4-((2-methoxyethoxy)methyl)-6-
-236-
SUBSTITUTE SHEET ( RULE 26 ) (trifluoromethyl)benzo[d]thiazol-2-amine (100 mg, 326.47 pmol, Example 13) in DCM (5 mL) was added EDCI (75 mg, 391.77 pmol) and HOBt (66 mg, 489.71 pmol). The mixture was stirred at 50 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 3- ((bis(2-methoxyethyl)amino)methyl)-5-chl oro-2 -hydroxy -N-(4- (2 -methoxy ethoxy)methyl)- 6-(trifluoromethyl)benzo[<7]thiazol-2-yl]benzamide (79.6 mg, 122.63 pmol, 37.56% yield, 98.98% purity, HC1) as a white solid. 1 H NMR (400 MHz, METHANOL-^) 8 ppm 8.14 (d, J= 2.45 Hz, 1 H), 8.10 (s, 1 H), 7.70 (s, 1 H), 7.56 (d, J= 2.45 Hz, 1 H), 4.88 (s, 2 H), 4.46 (s, 2 H), 3.64 - 3.72 (m, 6 H), 3.53 - 3 60 (m, 2 H), 3.36 - 3.44 (m, 4 H), 3 33 (s, 3 H), 3.20 (dt, J= 3.30, 1.53 Hz, 6 H). MS (ESI) [M+H]+ requires m/z 606.1 , found m/z 606.2.
Example 71
5-chloro-2-hydroxy-3-methyl-7V-(4-(tetrahydrofuran-2-yl)-6- (trifluoromethy l)b enzo \d] t h i azol -2-y l)b enzami de (71 )
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SUBSTITUTE SHEET ( RULE 26 ) [0527| To a solution of 4-bromo-6-(trifluorornethyl)benzo[d]thiazol-2-amine (2.6 g, 8.75 mmol) in DCM (60 mL) was added TEA (2.6 g, 26.25 mmol, 3.65 mL) and Boc O (4.7 g, 21.88 mmol, 5.03 mL) and DMAP (53 mg, 437.57 pmol). The mixture was stirred at 20 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by column chromatography (Si O2, petroleum ether/ethyl acetate = 1/0 to 100/1) to afford tert-butyl (4-bromo-6-(trifluoromethyl)benzo[</]thiazol-2- yl)carbamate (1.5 g, 1.77 mmol, 20.28% yield, 47% purity) as a white solid.
[0528] A mixture of tert-butyl (4-bromo-6-(trifluoromcthyl)bcnzo[t/]thiazol-2- yl)carbamate (1 g, 2.52 mmol), 2,3 -dihydrofuran (352 mg, 5.04 mmol, 380.70 pL), K2CO3 (1 g, 7.55 mmol), diacetoxypalladium (56 mg, 251.76 pmol) and PPhi (132 mg, 503.51 pmol) in DMF (10 mL) was degassed and purged with N2 3 times, then the mixture was stirred at 110 °C for 2 h under N2 atmosphere. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (40 mL x 3). The combined organic layer was washed with brine (100 mL x 3), dried over Na2SC>4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®;4 g SepaFlash® Silica Flash Column, eluent of 0-20% ethyl acetate/petroleum ether gradient @70 mL / min) to afford /crt-butyl (4-(2,5-dihydrofuran-2-yl)-6- (trifluoromethyl)benzo[rZ]thiazol-2-yl)carbamate (254 mg, 657.37 pmol, 26.11% yield) as yellow oil.
]0529] A mixture of tert-butyl (4-(2,5-dihydrofuran-2-yl)-6- (trifluorornethyl)benzo[ ]thiazol-2-yl)carbamate (0.2 g, 517.61 pmol), Pd/C (10 mg, 517.61 pmol, 10% purity), and NH3.H2O (145 mg, 1.04 mmol, 159.49 pL, 25% purity) in MeOH (4 mL), was degassed and purged with H2 3 times, then the mixture was stirred at 20 °C for 5 min under H (15 psi) atmosphere. The reaction mixture was filtered and concentrated under reduced pressure to afford tert-butyl (4-tetrahydrofuran-2-yl)-6- (trifluoromethyl)benzo[ ]thiazol-2-yl)carbamate (120 mg) as a white solid.
[0530] To a solution of tert-butyl (4-tetrahydrofuran-2-yl)-6- (trifluoromethyl)benzo[r/]thiazol-2-yl)carbamate (120 mg, 308.96 pmol) in DCM (2 mL) was added TFA (2 mL). The mixture was stirred at 20 °C for 1 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by column chromatography (SiCh, petroleum ether / ethyl acetate = 1/0 to 3/1) to afford 4- (tetrahydrofuran-2-yl)-6-(trifluoromethyl)benzo[r/]thiazol-2-amine (80 mg, 241.62 pmol, 78.21% yield, 87.071% purity) as a yellow solid.
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SUBSTITUTE SHEET ( RULE 26 ) [0531| To a solution of 4-(tetrahydrofuran-2-yl)-6- (trifluoromethyl)benzo[J]thiazol-2-amine (60 mg, 208.12 ymol), 5-chloro-2-hydroxy-3- methylbenzoic acid (42 mg, 228.94 pmol) in DCM (5 mb) was added EDCI (47 mg, 249.75 pmol) and HOBt (42 mg, 312.19 pmol). The mixture was stirred at 45 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (neutral condition) to afford 5-chloro-2-liydroxy-3-metliyl-/V-(4- (tetrahydrofuran-2-yl)-6-(trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (6.06 mg, 12.84 pmol, 6.17% yield, 96.78% purity) was obtained as a white solid. LH NMR (400 MHz, MF.THANOI.-Ji) 8 ppm 8.10 (s, 1 H), 7.85 (br s, 1 H), 7.70 (br s, 1 H), 7.26 (br s, 1 H), 5.60 (br s, 1 H), 4.23 (br d, J= 7.83 Hz, 1 H), 4.02 (br d, J= 7.34 Hz, 1 H), 2.65 (br s, 1 H), 2.24 (s, 3 H), 2.07 (br d, J = 6.85 Hz, 2 H), 1.81 (br s, 1 H). MS (ESI) [M+H]+ requires m/z 457.0, found m/z 457.1.
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SUBSTITUTE SHEET ( RULE 26 ) Example 72
5-chloro-3-((cz5-2,6-dimethylmorpholino)methyl)-2-hydroxy-JV-(4- (methylsulfonamidomethyl)-6-(trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (72)
[0532| To a solution of ZezZ-butyl (methylsulfonyl)carbamate (381 mg, 1.96 mmol) in MeCN (30 mb) was added CS2CO3 (1.9 g, 5.87 mmol). The mixture was stirred at 80 °C for 3 h, then ZcvZ-butyl A''-(4-(bromomethyl)-6-(trifluoromethyl)benzo[t/]thiazol-2-yl)- #-Zcz7-butoxycarbonyl carbarn ate (1 g, 1.96 mmol, Example 64) was added, the mixture was stirred at 80 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with water (5 mL) and extracted with ethyl acetate (7 mL x 3). The combined organic layer was washed with brine (20 mL), dried over NaiSCh, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®, 12 g SepaFlash® Silica Flash Column, Eluent of 0-10% ethylacetate/petroleum ether gradient @ 70 mL/min) to afford ZcvZ-butyl ('(2-((Zez'Z-
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SUBSTITUTE SHEET ( RULE 26 ) butoxy carbonyl)amino)-6-(trifluoromethyl)benzo[6?]thiazol-4- yl)methyl)(methylsulfonyl)carbamate (0.3 g, 481.77 pmol, 24.6% yield, 84.4% purity) as yellow oil.
|0533] To a solution of /cv7-butyl ((2-((/cv7-butoxycarbonyl)amino)-6- (trifluoromethyl)benzo[J]thiazol-4-yl)methyl)(methylsulfonyl)carbamate (0.3 g, 570.82 pmol) in DCM (3.0 mb) was added TFA (1.0 mL). The mixture was stirred at 20 °C for 2 h. The mixture was adjusted to pH 7-8 with sat. NaHCCh, the residue was diluted with water (10 mL) and extracted with DCM (5mL x 3). The combined organic layers were washed with brine (10 mL), dried over NaiSCL, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®, 4 g SepaFlash® Silica Flash Column, Eluent of 0-50% ethyl acetate/petroleum ether gradient @ 70 mL/min) to afford A-((2-amino-6-(trifluoromethyl)benzo[t/]thiazol-4- yl)methyl)methanesulfonamide (90 mg, 190.88 pmol, 33.4% yield, 69% purity) as a white solid.
10534| To a solution of A-((2-amino-6-(trifluoromethyl)benzo[t/]thiazol-4- yl)methyl)methanesulfonamide (30 mg, 92.21 pmol), 5-chloro-3-((cA-2,6- dimethylmorpholino)methyl)-2 -hydroxybenzoic acid (30 mg, 101.44 pmol, Example 64) in DCM (4.0 mL) was added EDCI (22 mg, 119.88 pmol) and HOBt (18 mg, 138.32 pmol). The mixture was stirred at 45 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 5-chloro-3-((cA-2,6-dimethylmorpholino)methyl)-2-hydroxy-A-(4- (methanesulfonamidomethyl )-6-(tri fluoromethyl )benzo[6/]thiazol-2-yl)benzamide (44.18 mg, 99.57% purity, HC1) as a white solid. 'H NMR (400 MHz, METHANOL-^) 8 ppm 8.26 - 8.32 (m, 2 H), 7.84 (s, 1 H), 7.75 (s, 1 H), 4.77 (s,2 H), 4.42 (s, 2 H), 3.90 (br s, 2 H), 3.44 (br d, J= 12.13 Hz, 2 H), 3.00 (s, 3 H), 2.84 (t, J= 11.69 Hz, 2 H), 1.25 (d, J= 6.17 Hz, 6 H); MS (ESI) [M+H]+ requires m/z 607.10, found m/z 607.1.
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SUBSTITUTE SHEET ( RULE 26 ) Example 73
5-chloro-2-hydroxy-3-methyl-A-(4-(methylsulfonamidomethyl)-6- (trifluoromethy l)b cnzo[t/] th i azol -2-y l)b enzami de (73 )
|0535] 5-chl oro-2 -hydroxy-3-methyl-/V-(4-(methylsulfonamidomethyl)-6-
(trifluoromethyl)benzo[t/]thiazol-2-yl)benzamide was made in a manner similar to Example 72. LCMS: MS (ESI) [M+H]+ requires m/z 494.01, found m/z 493.9. 'HNMR (400 MHz, DMSO-de) 8 ppm 8.47 (s, 1 H), 8.05 (br s, 1 H), 7.83 (s, 1 H), 7.77 (t, J = 6.17 Hz, 1 H), 7.50 (br s, 1 H), 4.67 (br d, J = 5.95 Hz, 2 H), 2.98 (s, 3 H) 2.23 (s, 3 H)
Example 74
3-((bis(2 -methoxy ethyl)amino)methyl)-5-chloro-2 -hydroxy -JV-(4- (methylsulfonamidomethyl)-6-(trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (74)
(0536] 3-((bis(2-methoxyethyl)amino)methyl)-5-chl oro-2 -hydroxy -7V-(4-
(methylsulfonamidomethyl)-6-(trifluorornethyl)benzo[</]thiazol-2-yl)benzamide was made in a manner similar to Example 72. LCMS: MS (ESI) [M+H]+ requires m/z 625.11, found m/z 625.1. 1HNMR (400 MHz, METHANOL-^) 8 ppm 8.26 (s, 2 H), 7.83 (s, 1 H), 7.71 (d, J = 2.21 Hz, 1 H), 4.75 (s, 2 H), 4.57 (s, 2 H), 3.75 - 3.80 (m, 4 H), 3.47 - 3.52 (m, 4 H), 3.42 (s, 6 H), 2.99 (s, 3 H)
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SUBSTITUTE SHEET ( RULE 26 ) Example 75
5-chloro-2-hydroxy-3-methyl-/V-(4-((2-(4-methylpiperazin-l-yl)ethoxy)methyl)-6- (trifluoromethy l)b cnzo[t/] th i azol -2-y l)b enzami de (75)
[0537| To a mixture of Ze/7- butyl 2V-(4-(bromomethyl)-6-
(trifluoromethyl)benzo[c/]thiazol-2-yl)-Af-Zez'Z-butoxycarbonylcarbamate (0.1 g, 195.56 pmol, Example 64), 2-(4-methylpiperazin-l-yl)ethan-l-ol (56 mg, 391.12 pmol) in THF (4.0 mL) was added t-BuOK (1 M, 782.24 pL, in TEIF) at 0 °C, then the mixture was stirred at 20 °C for 1 h. The residue was diluted with water (3 mL) and extracted with ethyl acetate (3 mL x 3). The combined organic layers were washed with brine (5 mL), dried over Na2SCU, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep- TLC (S1O2, DCM/MeOH = 10/1) to afford Ze/Z-butyl /V-Zc7'Z-butoxycarbonyl-A'f-(4-(2-(4- methylpiperazin-l-yl)ethoxymethyl)-6-(trifluoromethyl)benzo[J]thiazol-2-yl)carbamate (100 mg, 104.93 pmol, 53.6% yield, 60.3% purity) as yellow oil.
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SUBSTITUTE SHEET ( RULE 26 ) [0538| To a solution of /c77-butyl A-/c77-butoxycarbonyl-,V-(4-(2-(4- methylpiperazin-1 -yl)ethoxymethyl)-6-(tri fluoromethyl )benzo[r/]thiazol-2-yl)carbamate (0.1 g, 174.02 pmol) in DCM (2.0 mL) was added TFA (1 mL). The mixture was stirred at 20 °C for 1 h. The reaction mixture was pulled into sat. NaHCCh (10 mL), and then extracted with DCM (3 mL x 4). The combined organic layer was washed with brine (10 mL x 2), dried over NazSCL, fdtered and concentrated under reduced pressure to give 4-((2-(4- methylpiperazin-l-yl)ethoxy)methyl)-6-(trifluoromethyl)benzo[<7]thiazol-2-amine (50 mg, crude) as yellow oil.
[0539] A mixture of 4-((2-(4-methylpiperazin-l-yl)ethoxy)methyl)-6- (trifluoromethyl)benzo[r/]thiazol-2-amine (25 mg, 66.77 pmol), 5-chloro-2-hydroxy-3- methylbenzoic acid (14 mg, 80.12 pmol), EDCI (15 mg, 80.12 pmol), HOBt (13 mg, 100.15 pmol) in DCM (3.0 mL), was stirred at 45 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep- HPLC (HCL condition) to afford 5-chloro-2-hydroxy-3-methyl-A-(4-((2-(4-methylpiperazin- l-yl)ethoxy)methyl)-6-(trifluoromethyl)benzo[</]thiazol-2-yl)benzamide (19.1 mg, 99.49% purity, HC1 salt) as a white solid. 'H NMR (400 MHz, DMSO-<7y) 3 ppm 8.51 (br s, 1 H), 8.03 (br s, 1 H), 7.86 (s, 1 H), 7.49 (br s, 1 H), 4.99 (s, 2 H), 3.98 (br s, 2 H), 3.24 - 3.43 (m, 10 H), 2.80 (br s, 3 H), 2.24 (s, 3 H); MS (ESI) [M+H]+ requires m/z 543.14, found m/z
543.1
Example 76
5-chloro-3-((cA-2,6-dimethylmorpholino)methyl)-2-hydroxy-A-(4-((2-(4-methylpiperazin-l- yl)ethoxy)methyl)-6-(trifluoromethyl)benzo[rZ]thiazol-2-yl)benzamide (76)
[0540] 5-chloro-3-((cA-2,6-dimethylmorpholino)methyl)-2-hydroxy-A-(4-((2-(4- methylpiperazin-l-yl)ethoxy)methyl)-6-(trifluoromethyl)benzo[d]thiazol-2-yl)benzamide was
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SUBSTITUTE SHEET ( RULE 26 ) made in a manner similar to Example 75. LCMS: MS (ESI) [M+H]+ requires m/z 656.22, found m/z 656.3. 1HNMR (400 MHz, METHANOL-ch) 8 ppm 8.28 - 8.35 (m, 2 H), 7.87 (s, 1 H), 7.79 (d, J = 2.43 Hz, 1 H), 5.09 (s, 2 H), 4.45 (s, 2 H), 4.06 - 4.12 (m, 2 H), 3.54 - 4.05 (m, 12 H), 3.47 (br d, J = 11.91 Hz, 2 H), 3.00 (s, 3 H), 2.87 (t, J = 11.80 Hz, 2 H),1.24 (d, J = 6.17 Hz, 6 H)
Example 77
3-((bis(2-methoxyethyl)amino)methyl)-5-chloro-2-hydroxy-7V-(4-((2-(4-methylpiperazin-l- yl)ethoxy)methyl)-6-('tri fluoromethyl )benzo[<7]thiazol-2-yl)benzamide (77)
[0541] 3-((bis(2-methoxyethyl)amino)methyl)-5-chloro-2-hydroxy-Ar-(4-((2-(4- methylpiperazin-l-yl)ethoxy)methyl)-6-(trifluoromethyl)benzo[d]thiazol-2-yl)benzamide was made in a manner similar to Example 75. LCMS: MS (ESI) [M+H]+ requires m/z 674.23, found m/z 674.3. LHNMR (400 MHz, METHANOL-^) 8 ppm 8.32 (d, J= 2.43 Hz, 1 H), 8.29 (s, 1 H), 7.87 (s, 1 H), 7.75 (d, J= 2.65 Hz, 1 H), 5.09 (s, 2 H), 4.61 (s, 2 H), 4.06 - 4.13 (m, 2 H), 3.60 - 4.02 (m, 15 H), 3.52 (br d, J= 16.98 Hz, 4 H), 3.42 (s, 6 H), 3.00 (s, 3 H)
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SUBSTITUTE SHEET ( RULE 26 ) Example 78
5-chloro-2-hydroxy-3 -methyl -A'-(4-(( 4-methyl pi perazi n-1 -y I (methyl )-6- (trifluoromethy l)b cnzo[t/] th i azol -2-y l)b enzami de (78)
(0542| To a solution of 1 -methylpiperazine (29 mg, 293.34 pmol, 32.54 pL) in MeCN (2.0 mL) was added K2CO3 (81 mg, 586.68 pmol) and the mixture was stirred at 0 °C for 10 minutes, then tert-butyl 7V-(4-(b romom ethyl )-6-(trifluorom ethyl (benzo[<7]thi azol -2-yl )- A-ter/-butoxycarbonylcarbamate (0.1 g, 195.56 pmol, Example 64) was added, and the mixture was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with water (2 mL) and extracted with ethyl acetate (3 mL x 2). The combined organic layer was washed with brine (5 mL), dried over Na2SC>4, fdtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, DCM/MeOH = 10/1) to afford terLbutyl N-tert- butoxycarbonyl -/V-(4-((4-mcthylpipcrazin- l -yl (methyl )-6-(trifluorom ethyl )bcnzo[6/]thiazol-2- yl)carbamate (100 mg, 144.93 pmol, 74.1% yield, 76.9% purity) as yellow oil.
|0543] To a solution of tert-butyl A'-/c/7-butoxycarbonyl-.V-(4-((4- methylpiperazin-l-yl)methyl)-6-(trifluoromethyl)benzo|p(]thiazol-2-yl)carbamate (100 mg, 188.47 pmol) in DCM (2.0 mL) was added TFA (1 mL). The mixture was stirred at 20 °C for 1 h. The reaction mixture was pulled into sat. NaHCO, (10 mL), and then extracted with
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SUBSTITUTE SHEET ( RULE 26 ) DCM (3 mL * 4). The combined organic layer was washed with brine (10 mb x 2), dried over NazSCM, filtered and concentrated under reduced pressure to give 4-((4- methylpiperazin-l-yl)methyl)-6-(trifluoromethyl)benzo[<7]thiazol-2-amine (50 mg, crude) as yellow oil.
[0544j A mixture of 4-((4-methylpiperazin- 1 -yl)methyl)-6- (trifluoromethyl)benzo[r/]thiazol-2-amine (25 mg, 75.67 pmol), 5-chloro-2-hydroxy-3- methylbenzoic acid (16 mg, 90.81 pmol), HOBt (15 mg, 113.51 pmol), EDCI (17 mg, 90.81 pmol) in DCM (3 mL), was stirred at 45 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent The residue was purified by prep-HPLC (HC1 condition) to afford 5-chloro-2-hydroxy-3-methyl-7V-(4-((4-methylpiperazin-l-yl)methyl)-6- (trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (15 mg, 100% purity, HC1) as a white solid. XH NMR (400 MHz, DMSO-t76) 3 ppm 8.61 (br s, 1 H), 8.17 (br s, 1 H), 8.08 (br s, 1 H), 7.50 (br s, 1 H), 4.68 (br s, 2 H), 3.34 - 3.51 (m, 8 H), 2.79 (br s, 3 H), 2.24 (s, 3 H); MS (ESI) [M+H]+ requires m/z 499.11, found m/z 499.1
Example 79
5-chloro-3-((czs-2,6-dimethylmorpholino)methyl)-2-hydroxy-A-(4-((4-methylpiperazin-l- yl)methyl)-6-(trifluoromethyl)benzo[<7]thiazol-2-yl)benzamide (79)
[0545] 5-chloro-3-((cA-2,6-dimethylmorpholino)methyl)-2-hydroxy-JV-(4-((4- methylpiperazin-l-yl)methyl)-6-(trifluoromethyl)benzo[d]thiazol-2-yl)benzamide was made in a manner similar to Example 78. LCMS: MS (ESI) [M+H]+ requires m/z 612.19, found m/z 612.2. 1HNMR (400 MHz, METHANOL-^) 5 ppm 8.47 - 8.56 (m, 2 H), 8.02 (s, 1 H), 7.86 (d, J= 2.43 Hz, 1 H), 4.89 (br s, 2 H), 4.45 (s, 2 H), 3.90 (br d, J= 6.17 Hz, 2 H), 3.74 (br s, 8 H), 3.46 (br d, J= 11.69 Hz, 2 H), 3.01 (s, 3 H), 2.80 - 2.92 (m, 2 H), 1.24 (d, J =639 Hz, 6 H)
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SUBSTITUTE SHEET ( RULE 26 ) Example 80
3-((bis(2 -methoxy ethyl)amino)methyl)-5-chl oro-2 -hydroxy -A-(4-((4-methylpiperazin-l - yl)methyl)-6-(trifluoromethyl)benzo[t/]thiazol-2-yl)benzamide (80)
[0546] 3-((bis(2 -methoxyethyl)amino)methyl)-5-chl oro-2 -hydroxy -JV-(4-((4- methylpiperazin-l-yl)methyl)-6-(trifluoromethyl)benzo[d]thiazol-2-yl)benzamide was made in a manner similar to Example 78. LCMS: MS (ESI) [M+H]+ requires m/z 630.21, found m/z 630.2. 1HNMR (400 MHz, METHANOL-^) 8 ppm 8.41 - 8.50 (m, 2 H), 7.98 (s, 1 H), 7.80 (d, J= 2.43 Hz, 1 H), 4.71 (br s, 2 H), 4.61 (s, 2 H),3.79 (t, J= 4.96 Hz, 4 H), 3.47 - 3.73 (m, 12 H), 3.42 (s, 6 H), 2.98 (s, 3 H)
Example 81
5-chloro-2-hydroxy-3-((4-methylpiperazin-l-yl)methyl)-/V-(4-((4-methylpiperazin-l- yl)methyl)-6-(trifluoromethyl)benzo[<7]thiazol-2-yl)benzamide(81)
[0547] 5 -chi oro-2 -hy droxy-3 -((4-methylpiperazin- 1 -yl)methyl)-A'-(4-((4- methylpiperazin-l-yl)methyl)-6-(trifluoromethyl)benzo[6?]thiazol-2-yl)benzamide was made in a manner similar to Example 78. LCMS: MS (ESI) [M+H]+ requires m/z 597.19, found m/z 597.2. XHNMR (400 MHz, DMSO-fifc) 5 ppm 11.87 (br s, 1 H), 8.61 (s, 1 H), 8.20 (br s, 1 H), 8.13 (br s, 1 H), 7.88 (br d, J= 1.76 Hz, 1 H), 4.74 (br s, 2 H), 4.36 (br s, 2 H), 3.61 (br d, J = 7.28 Hz, 8 H), 3.44 (br s, 8 H), 2.82 (br d, J = 12.13 Hz, 6 H)
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SUBSTITUTE SHEET ( RULE 26 ) Example 82
5-chloro-/V-(4-(((4-chlorobenzyl )oxy)methyl)-6-(tri fluoromethyl )benzo[c/]thiazol-2-yl)-2- hydroxy-3 -methylbenzamide (82)
|0548] To a stirred solution of (4-chlorophenyl)methanol (83 mg, 586.68 pmol, 6.86 pL) in DMF (4.0 mL) at -10 °C, was added NaH (117 mg, 2.93 mmol, 60% purity). After 1 h, Ze/7-butyl A-(4-(bromomethyl)-6-(trifluoromethyl)benzo[<7]thiazol-2-yl)-A-ZerZ- butoxycarbonylcarbamate (200 mg, 293.34 pmol, Example 64) was added. Stirring was continued at -10 °C for 1 h. The reaction mixture was quenched by addition sat. NH4CI (5 mL), and then diluted with water (5 mL) and extracted with DCM (5 mL x3). The combined organic layer was washed with brine (10 mL), dried over NaiSCL, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCh, petroleum ether/ethyl acetate = 3/1) to afford ZcvZ-butyl (4-(((4-chlorobenzyl)oxy)methyl)-6- (trifluoromethyl)benzo[r/]thiazol-2-yl)carbamate (72 mg, 86.70 pmol, 29.5% yield, 69% purity) as a yellow solid.
|0549| To a solution of ZcvZ-butyl A'-Zc/'Z-butoxycarbonyl-A-[4-[(4- chlorophenyl)methoxymethyl]-6- (trifluoromethyl)-l,3-benzothiazol-2-yl]carbamate (70 mg, 122.16 pmol) in DCM (2.0 mL) was added TFA (1 mL). The mixture was stirred at 20 °C for 5 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The
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SUBSTITUTE SHEET ( RULE 26 ) residue was diluted with H2O (3 mL) and extracted with DCM (2 mL * 2). The combined organic layer was washed with brine (5 mL), dried over NazSCE, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCh, petroleum ether/ethyl acetate = 3/1) to afford 4-(((4-chlorobenzyl)oxy)methyl)-6- (trifluoromethyl)benzo[J]thiazol-2-amine (40 mg, 58.16 pmol, 47.6% yield, 54.2% purity) as a yellow solid.
[0550] To a solution of 4-(((4-chlorobenzyl)oxy)methyl)-6- (trifluoromethyl)benzo[d]thiazol-2-amine (40 mg, 107.30 pmol), 5-chloro-2-hydroxy-3- methylbenzoic acid (40 mg, 214.60 pmol) in DCM (4 0 mL) was added EDCI (24 mg, 128.76 pmol) and HOBt (21 mg, 160.95 pmol). The mixture was stirred at 50 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 5-chloro-7V-(4-(((4- chlorobenzyl)oxy)methyl)-6-(trifluoromethyl)benzo[J]thiazol-2-yl)-2-hydroxy-3- methylbenzamide (8.4 mg, 15.61 pmol, 14.5% yield, 100% purity) as a white solid. LH NMR (400 MHz, METHANOL-^) 8 ppm 8.22 (s, 1 H), 7.95 (d, J= 2.45 Hz, 1 H), 7.79 (s, 1 H), 7.33 - 7.42 (m, 5 H), 5.05 (s, 2 H), 4.69 (s, 2 H), 2.28 (s, 3 H); MS (ESI) [M+H]+ requires m/z 541.03, found m/z 541.1.
Example 83
5-chloro-JV-(4-(((4-fluorobenzyl)oxy)methyl)-6-(trifluoromethyl)benzo[</]thiazol-2-yl)-2- hydroxy-3 -methylbenzamide (83)
[0551] 5 -chloro-/V-(4-(((4-fluorobenzyl)oxy)methyl)-6-
(trifluoromethyl)benzo[t/]thiazol -2 -yl)-2 -hydroxy-3 -methylbenzamide was made in a manner similar to Example 82. LCMS: MS (ESI) [M+H]+ requires m/z 525.06, found m/z 525.1.
1HNMR (400 MHz, DMSO-r/r,) 8 ppm 8.47 (s, 1 H), 8.03 (br s, 1 H), 7.78 (s, 1 H), 7.36 - 7.52
(m, 3 H), 7.19 (t, J= 8.80 Hz, 2 H), 4.98 (s, 2 H),4.66 (s, 2 H),2.22 (s, 3 H)
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SUBSTITUTE SHEET ( RULE 26 ) Example 84
Ar-(4-((benzyloxy)methyl)-6-(trifluoromethyl)benzo[t/]thiazol-2 -yl)-5-chl oro-2 -hydroxy-3- methylbenzamide (84)
[0552| 7V-(4-((benzyloxy)methyl)-6-(trifluoromethyl)benzo[</]thiazol-2-yl)-5- chloro-2-hydroxy-3 -methylbenzamide was made in a manner similar to Example 82. LCMS: MS (ESI) [M+H]+ requires m/z 507.07, found m/z 507.2. 1HNMR (400 MHz, DMSO-d6) 8 ppm 8.47 (s, 1 H), 8.04 (br s, 1 H), 7.80 (s, 1 H),7.48 (br s, 1 H), 7.35 - 7.43 (m, 4 H), 7.28 - 7.34 (m, 1 H), 4.99 (s, 2 H),4.69 (s, 2 H), 2.22 (s, 3 H)
Example 85
5-chloro-2-hydroxy-3-methyl-jV-(4-(((4-methylbenzyl)oxy)methyl)-6-
(trifluoromethyl)benzo[rZ]thiazol-2-yl)benzamide (85)
|0553] To a solution of /cvV-butyl jV-(4-(bromomethyl)-6-
(trifluoromethyl)benzo[r/]thiazol-2-yl)-Ar-ter/-butoxycarbonylcarbamate (0.2 g, 391.12 pmol,
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SUBSTITUTE SHEET ( RULE 26 ) Example 64) and /?-tolylmethanol (95 mg, 782.24 pmol, 63.25 pL) in THF (5 mL) was added t-BuOK, (1 M, 1.56 mL, THF), at 0 °C, then the mixture was stirred at 20 °C for 1 h. The residue was diluted with water (5 mL) and extracted with ethyl acetate (3 mL x 2). The combined organic layer was washed with brine (3 mL x 2), dried over Na2SC>4, filtered and concentrated under reduced pressure to give tert-butyl (4-(((4-methylbenzyl)oxy)methyl)-6- (trifluoromethyl)benzo[c/]thiazol-2-yl)carbamate (150 mg, crude) as yellow oil.
[0554] To a solution of tert-butyl (4-(((4-methylbenzyl)oxy)methyl)-6- (trifluoromethyl)benzo[ ]thiazol-2-yl)carbamate (150 mg, 331.50 pmol) in DCM (2.0 mL) was added TFA (1.0 mL). The mixture was stirred at 30 °C for 1 h. The reaction mixture was put into sat. NaHCCh (10 mL) and extracted with DCM (5 mL x 2). The combined organic layer was washed with brine (20 mL x 2), dried over ISfeSCL, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep- TLC ( Si O2, petroleum ether/ethyl acetate = 1/1) to afford 4-(((4-methylbenzyl)oxy)methyl)- 6-(trifluoromethyl)benzo[<7]thiazol-2-amine (50 mg, 76.62 pmol, 23.11% yield, 54% purity) as a yellow solid.
[0555] To a solution of 4-(((4-methylbenzyl)oxy)methyl)-6- (trifluoromethyl)benzo[d]thiazol-2-amine (30 mg, 85.14 pmol), 5-chloro-2-hydroxy-3- methylbenzoic acid (19 mg, 102.16 pmol) in DCM (2.0 mL) was added EDCI (19 mg, 102.16 pmol) and HOBt (17 mg, 127.71 pmol). The mixture was stirred at 45 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (neutral condition) to afford 5-chloro-2-hydroxy-3-methyl-/V-(4- (((4-methylbenzyl)oxy)m ethyl )-6-(trifluorom ethyl )benzo[<7]thiazol-2-yl (benzamide (10.9 mg, 97.23% purity) as a white solid. 'H NMR (400 MHz, DMSO-fifc) 5 ppm 8.46 (br s, 1 H), 8.03 (br s, 1 H), 7.77 (br s, 1 H), 7.47 (br s, 1H), 7.28 (br d, J= 7.50 Hz, 2 H), 7.18 (br d, J= 7.50 Hz, 2 H), 4.96 (br s, 2 H), 4.63 (s, 2 H), 2.30 (s, 3 H), 2.22 (s, 3 H); MS (ESI) [M+H]+ requires m/z 521.08, found m/z 521.2.
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SUBSTITUTE SHEET ( RULE 26 ) Example 86
5-chloro-/V-(4-((2-(dimethylamino)ethoxy)methyl)-6-(trifluoromethyl)benzo[</]thiazol-2-yl)- 2-hydroxy-3 -methylbenzamide (86)
(0556] 5-chloro-Ar-(4-((2-(dimethylamino)ethoxy)methyl)-6- (trifluoromethyl)benzo[<7]thiazol -2 -yl)-2 -hydroxy-3 -methylbenzamide was made in a manner similar to Example 85. LCMS: MS (ESI) [M+H]+ requires m/z 488.09, found m/z 488.0. XHNMR (400 MHz, DMSO-ds) 5 ppm 9.72 (br s, 1 H), 8 19 (s, 1 H), 7.69 (d, J = 2.65 Hz, 1 H), 7.62 (s, 1 H), 7.21 (d, J = 2.87 Hz, 1 H), 4.97 (s, 2 H),3.90 (t, J= 5.07 Hz, 2 H), 3.42 (br t, J = 4.96 Hz, 2 H), 2.89 (s, 6 H), 2.16 (s, 3 H)
Example 87
5-chloro-2-hydroxy-3-methyl-7V-(4-((thiazol-2-ylmethoxy)methyl)-6- (trifluoromethy l)b enzo[<7] th i azol -2-y l)b enzami de (87)
|0557] 5-chl oro-2 -hy droxy-3-methyl-/V-(4-((thi azol-2-ylmethoxy )methyl)-6-
(trifluoromethyl)benzo[d]thiazol-2-yl)benzamide was made in a manner similar to Example 85. LCMS: MS (ESI) [M+H]+ requires m/z 514.02, found m/z 513.9. ‘HNMR (400 MHz, DMSO-ds) 5 ppm 8.49 (s, 1 H), 8.04 (br s, 1 H), 7.82 (d, J = 3.09 Hz, 2 H), 7.76 (d, J = 3.31 Hz, 1 H), 7.48 (br s, 1 H), 5.11 (s, 2 H), 5.00 (s, 2 H), 2.22 (s, 3 H)
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SUBSTITUTE SHEET ( RULE 26 ) Example 88
5-chloro-2-hydroxy-JV-(4-((2-hydroxyethoxy)methyl)-6-(trifluoromethyl)benzo[</]thiazol-2- yl)-3 -methylbenzamide (88)
[0558] 5-chl oro-2 -hydroxy -7/-(4-((2-hydroxy ethoxy)methyl)-6-
(trifluoromethyl)benzo[c/]thiazol-2-yl)-3 -methyl benzamide was made in a manner similar to Example 85.LCMS: MS (ESI) [M+H]+ requires m/z 461.05, found m/z 460.9. LHNMR (400 MHz, DMSO-fifc) 5 ppm 8.46 (s, 1 H), 8.03 (br s, 1 H), 7.83 (s, 1 H), 7.48 (br s, 1 H), 4.96 (br s, 2 H), 3.61 (s, 4 H), 2.23 (s, 3 H)
Example 89
5-chloro-2-hydroxy-7V-(4-(((4-methoxybenzyl)oxy)methyl)-6- (trifluoromethyl)benzo[<7]thiazol-2-yl)-3 -methylbenzamide (89)
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SUBSTITUTE SHEET ( RULE 26 ) [0559| To a solution of te/7-butyl A'-(4-(bromomethyl(-6- (trifluoromethyl)benzo[d]thiazol-2-yl)-/V-te/7-butoxycarbonylcarbamate (0.4 g, 782.24 pmol, Example 64) and (4-methoxyphenyl)methanol (140 mg, 1.02 mmol, 126.58 pL) in TEIF (10 0 mL) was added tBuOK, (1 M, 3.13 mL, THF) at 0 °C, then the mixture was stirred at 20 °C for 12 h. The residue was diluted with water (20 mL) and extracted with ethyl acetate (10 mL x 2). The combined organic layer was washed with brine (20 mL x 2), dried over NazSCU, fdtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0~10% ethyl acetate/petroleum ether gradient @ 70 mL / min) to afford tert-butyl (4-((('4-methoxybenzyl(oxy(methyl(-6-(trifluoromethyl(benzo[<7]thiazol -2-yl (carbamate (170 mg, 362.87 pmol, 46.39% yield) as yellow oil.
|0560] A solution of tert-butyl (4-(((4-methoxybenzyl)oxy)methyl)-6- (trifluoromethyl)benzo[t/]thiazol-2-yl)carbamate (100 mg, 213.45 pmol) in 1, 1,1, 3,3,3- hexafluoropropan-2-ol (3.0 mL) was stirred at 80 °C for 1.5 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep- TLC (SiCb, petroleum ether/ethyl acetate = 3/1) to afford 4-(((4-methoxybenzyl)oxy)methyl)- 6-(trifluoromethyl)benzo[<7]thiazol-2-amine (70 mg, 190.02 pmol, 89.0% yield) as a yellow solid.
[0561] To a solution of 4-(((4-methoxybenzyl)oxy)methyl)-6- (trifluoromethyl)benzo[d]thiazol-2-amine (70 mg, 190.02 pmol), and 5-chl oro-2 -hydroxy-3- methylbenzoic acid (35 mg, 190.02 pmol) in DCM (3.0 mL) was added EDCI (43 mg, 228.03 pmol) and HOBt (38 mg, 285.04 pmol). The mixture was stirred at 45 °C for 3 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (neutral condition) to afford 5-chloro-2-hydroxy-A-(4-(((4- methoxybenzyl)oxy)methyl)-6-(trifluoromethyl)benzo[7/]thiazol-2-yl)-3-methylbenzamide (25 mg, 47.92 pmol, 25.2% yield, 100% purity) as a yellow solid. 'H WiR (400 MHz, DMSO-tA) 5 ppm 8.45 (s, 1 H), 8.03 (br s, 1 H), 7.76 (s, 1 H), 7.47 (br s, 1 H), 7.31 (d, J = 8.31 Hz, 2 H), 6.92 (d, J = 8.80 Hz, 2 H), 4.94 (s, 2 H), 4.60 (s, 2 H), 3.74 (s, 3 H), 2.22 (s, 3 H); MS (ESI) [M+H]+ requires m/z 537.08, found m/z 537.0
Example 90
5-chl oro-2-hydroxy-3-methyl-A-(4-(propionamidomethyl)-6- (trifluoromcthyl)bcnzo[t/]thiazol-2-yl (benzamide (90)
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SUBSTITUTE SHEET ( RULE 26 )
[0562] To a solution of zc/7-butyl A'-(4-(bromomethyl)-6- (trifluoromethyl)benzo[d]thiazol-2-yl)-jV-tert-butoxycarbonylcarbamate (300 mg, 586.68 pmol, Example 64) in DMF (5.0 mL) was added potassium l,3-dioxoisoindolin-2-ide) (163 mg, 880.02 pmol). The mixture was stirred at 25 °C for 12 h. The reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (5 mL x 3). The combined organic layer was washed with brine (15 mL x 2), dried over ISfeSCL, fdtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® Silica Flash Column, eluent of 0-10% ethyl acetate/petroleum ether gradient @ 70 mL/min) to afford Zc/Z-butyl N-tert- butoxycarbonyl-JV-(4-((l,3-dioxoisoindolin-2-yl)methyl)-6-(trifluoromethyl)benzo[6/]thiazol- 2-yl)carbamate (280 mg, 380.07 pmol, 64.78% yield, 78.4% purity) as a white solid.
[0563[ To a suspension of ZcvZ-butyl A-Zc77-butoxycarbonyl-;V-(4-(( l ,3- dioxoisoindolin-2-yl)methyl)-6-(trifluoromethyl)benzo[<7]thiazol-2-yl)carbamate (280 mg, 484.79 pmol) in EtOH (5.0 mL) was added N2H4.H2O (49 mg, 969.58 pmol, 48.09 pL, 98% purity). The mixture was stirred at 25 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with DCM (5 mL), then filtered and concentrated under reduced pressure to give Zc77-butyl (4-(aminomethyl)-6- (trifluoromethyl)benzo[c/]thiazol -2-yl (carbamate (150 mg, crude) as a white solid.
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SUBSTITUTE SHEET ( RULE 26 ) [0564| To a solution of t terZ-butyl (4-(aminomethyl)-6- (trifluoromethyl)benzo[J]thiazol-2-yl)carbamate (100 mg, 287.89 pmol) in DCM (4 mL) was added TEA (43 mg, 431.83 pmol, 60.11 uL), then propanoyl chloride (31 mg, 345.47 pmol, 31 ul) was added at 0°C. The resulting mixture was stirred at 20°C for 1 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-TLC (SiCh, petroleum ether/ethyl acetate 1/1) to afford tert-butyl (4- (propionamidomethyl)-6-(trifhioromethyl)benzo[d]thiazol-2-yl)carbamate (70 mg, 143.85 pmol, 49.97% yield, 82.9% purity) as a white solid.
[0565] To a solution of tert-butyl (4-(propionamidomethyl)-6- (trifluoromethyl)benzo[4/]thiazol-2-yl)carbamate (70 mg, 173.52pmol) in DCM (2.0 mL) was added TFA (0.5 mL). The mixture was stirred at 20 °C for 1 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with sat NaHCCh (5 mL) and extracted with DCM (3 mL x 3). The combined organic layers were washed with brine (5 mL x 3), dried over NazSCL, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/ethyl acetate = 0/1) to afford A-((2-amino-6-(trifluoromethyl)benzo[<7]thiazol-4- yl)methyl)propionamide (40 mg, 105.51 pmol, 60.8% yield, 80% purity) as a white solid.
[0566] To a solution of A-((2-amino-6-(trifluoromethyl)benzo[6(]thiazol-4- yl)methyl)propionam (25 mg, 82.43 pmol), and 5-chloro-2-hydroxy-3-methylbenzoic acid (21 mg, 115.40 pmol) in DCM (3.0 mL) was added EDCI (18 mg, 98.91 pmol) and HOBt (16 mg, 123.64 pmol). The mixture was stirred at 45 °C for 4 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep- HPLC (neutral condition) to afford 5-chloro-2-hydroxy-3-methyl-A-(4- (propionamidomethyl)-6-(trifluoromethyl)benzo[t(]thiazol-2-yl)benzamide (9.4 mg, 19.76 pmol, 23.9% yield, 99.18% purity) as a white solid. TH NMR (400 MHz, DMSO-^) 5 ppm 8.48 (br t, J= 5.62 Hz, 1 H), 8.42 (s, 1 H), 8.03 (br s, 1 H), 7.58 (s, 1 H), 7.48 (br s, 1 H), 4.72 (br s, 2 H), 2.19 - 2.26 (m, 5 H), 1.06 (t, J= 7.61 Hz, 3 H); MS (ESI) [M+H]+ requires m/z 472.06, found m/z 471.9
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SUBSTITUTE SHEET ( RULE 26 ) Example 91
5-chloro-2-hydroxy-3-methyl-/V-(4-((2-(methylsulfonamido)ethoxy)methyl)-6- (trifluoromethy l)b cnzo[t/] th i azol -2-y l)b enzami de (91 )
10567] To a mixture of ethylene glycol (21.3 g, 343.92 mmol, 19.23 mb) and pyridine (20.0 mL) was added toT-butylchlorodiphenylsilane (10.7 g, 38.93 mmol, 10 mL), the mixture was stirred at 25 °C for 1 h. The reaction mixture was diluted with ethyl acetate (120 mL) and poured in water (60 mL), the layers were separated and the organic phase was washed with aq. NaHCCh (60 mL, Wt 10%) and water (60 mL), dried over Na?SC>4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCh, petroleum ether/ethyl acetate = 1/0 to 10/1) to afford 2((tert- butyldiphenylsilyl)oxy)ethan-l-ol (9.8 g, 26.09 mmol, 67.03% yield, 80% purity) as yellow
-258-
SUBSTITUTE SHEET ( RULE 26 ) oil. 'H NMR (400 MHz, CHLOROFORM- ) 5 ppm 7.62 - 7.75 (m, 4 H), 7.29 - 7.52 (m, 6 H), 3.75 - 3.81 (m, 2 H), 3.66 - 3.72 (m, 2 H), 1.06 - 1.09 (m, 9 H)
|0568] To a solution of 2((tert-butyldiphenylsilyl)oxy)ethan-l-ol (1.7 g, 5.87 mmol), tert-butyl A-(4-(bromomethyl)-6-(trifluoromethyl)benzo[d]thiazol-2-yl)-JV-tert- butoxycarbonylcarbamate (1.5 g, 2.93 mmol, Example 64) in THF (60 mL) was added t- BuOK (1 M, 11.73 mL, in THF), at 0 °C, then the mixture was stirred at 20 °C for 1 h. The residue was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 2). The combined organic layer was washed with brine (20 mL), dried over NazSC , filtered and concentrated under reduced pressure to give tert-butyl A-tert-butoxycarbonyl-(4-((2-(tert- butyldiphenylsilyl)oxy)ethoxymethyl)-6-(trifluoromethyl)benzo[d]thiazol-2-yl)carbamate (2 g, crude) as yellow oil.
|0569] tert-butyl A-tert-butoxycarbonyl-(4-((2-(tert- butyldiphenylsilyl)oxy)ethoxymethyl)-6-(trifluoromethyl)benzo[d]thiazol-2-yl)carbamate (1 g, 1.37 mmol) was dissolved in THF (10 mL) and cooled to 0 °C then treated with TBAF (1 M, 2.74 mL). The resulting mixture was stirred at 0 °C for 5 minutes, then warmed to 20 °C and stirred for 12 h. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic layer was washed with brine (10 mL x 3), dried over NazSCL. filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0-50% ethyl acetate/petroleum ether gradient @ 70 mL/min) to afford tert-butyl (4-((2 -hydroxyethoxy )methyl)-6-(trifluoromethyl)benzo[6 ]thiazol -2- yl)carbamate (500 mg, 90.7% purity) as a white solid.
[0570] A solution of tert-butyl (methylsulfonyl)carbamate (29 mg, 152.28 pmol) in THF (3 mL) was added PPhz (79 mg, 304.56 pmol). The solution was stirred under N2 and the tert-butyl tert-butyl 7V-tert-butoxycarbonyl-zV-[4-(2-hydroxyethoxymethyl)-6- (trifluoromethyl)-l,3-benzothiazol-2-yl]carbamate (50 mg, 101.52 pmol) was added followed by DIAL) (51 mg, 253.80 pmol, 49.35 pL). The mixture was stirred at 20 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-TLC (Si O?, petroleum ether/ethyl acetate = 3/1) to afford tert-butyl (2- ((2-((tert-butoxycarbonyl)amino)-6-(trifluoromethyl)benzo[</]thiazol-4- yl)methoxy)ethyl)(methylsulfonyl)carbamate (60 mg, 75.88 pmol, 74.74% yield, 84.7% purity) as a white solid.
[0571] To a solution of tert-butyl (2-((2-((tert-butoxycarbonyl)amino)-6- (trifl uoromethyl)benzo[< ]thiazol -4-yl )methoxy)ethyl)(methyl sulfonyl (carbamate (60 mg,
-259-
SUBSTITUTE SHEET ( RULE 26 ) 89.59 pmol) in DCM (2.0 mL) was added TFA (1 mL). The mixture was stirred at 25 °C for 3 h. The reaction mixture was poured into sat NaHCOs (5 mL), the pH was adjusted with sat. NaHCChto around 9 and extracted with DCM (3 mL x 3). The combined organic layer was washed with brine (5 mL x 3), dried over NazSCU, filtered and concentrated under reduced pressure to give A-(2-((2-amino-6-(trifluoromethyl)benzo[</]thiazol-4- yl)methoxy)ethyl)methanesulfonamide (30 mg, crude) as a yellow solid.
[0572] To a solution of A-(2-((2-amino-6-(trifhjoromethyl)benzo[d]thiazol-4- yl)methoxy)ethyl)methanesulfonamide (30 mg, 81.22 pmol), 5-chloro-2-hydroxy-3- methylbenzoic acid (22 mg, 121.82 pmol) in DCM (3 0 mL) was added EDCI (18 mg, 97.46 pmol) and HOBt (16 mg, 121.82 pmol). The mixture was stirred at 45 °C for 5 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 5-chloro-2-hydroxy-3-methyl-,V-(4-((2- (methylsulfonamido)ethoxy)methyl)-6-(tri fluoromethyl )benzo[<7]thiazol-2-yl)benzamide (4.1 mg, 7.70 pmol, 9.4% yield, 100% purity) as a white solid. 'H NMR (400 MHz, DMSO-c4>) <5 ppm 8.47 (s, 1 H), 8.02 (br s, 1 H), 7.83 (s, 1 H), 7.48 (br s, 1 H), 7.12 - 7.23 (m, 1 H), 4.96 (br s, 2 H), 3.64 (br t, J= 5.73 Hz, 2 H), 3.13 - 3.27 (m, 2 H), 2.91 (s, 3 H), 2.23 (s, 3 H); MS (ESI) [M+H]+ requires m/z 538.04, found m/z 538.0.
Example 92
5-chloro-3-((czs-2,6-dimethylmorpholino)methyl)-2 -hydroxy -7V-(4-((2- (methylsulfonamido)ethoxy)methyl)-6-(trifluoromethyl)benzo[z7]thiazol-2-yl)benzamide (92)
[0573] 5-chloro-3-((c7.s-2.6-dimethylmorpholino)methyl)-2-hydroxy-/V-(4-((2- (methylsulfonamido)ethoxy)methyl)-6-(trifluoromethyl)benzo[</]thiazol-2-yl)benzamide was made in a manner similar to Example 91. LCMS: MS (ESI) [M+H]+ requires m/z 651.12, found m/z 651.2. 1HNMR (400 MHz, DMSO4) 8 ppm 8.36 (s, 1 H), 7.87 (br s, 1 H), 7.74
-260-
SUBSTITUTE SHEET ( RULE 26 ) (s, 1 H), 7.46 (br s, 1 H), 7.17 (t, J= 5.84 Hz, 1 H), 4.93 (s, 2 H), 4.25 (s, 2 H), 3.90 (br dd, J = 9.81, 6.73 Hz, 2 H), 3.64 (t, J= 5.73 Hz, 2 H), 3.32 (br d, J= 12.13 Hz, 2 H), 3.20 (q, J = 5.59 Hz, 2 H), 2.91 (s, 3 H), 2.63 - 2.74 (m, 2 H),1.12 (d, J= 6 17 Hz, 6 H)
Example 93
3-((bis(2-methoxyethyl)amino)methyl)-5-chloro-2-hydroxy-JV-(4-((2-
(methylsulfonamido)ethoxy)methyl)-6-(trifluoromethyl)benzo[</]thiazol-2-yl)benzamide (93)
[0574 | 3-((bis(2-methoxyethyl)amino)methyl)-5-chl oro-2 -hydroxy -/V-(4-((2- (methylsulfonamido)ethoxy)methyl)-6-(trifluoromethyl)benzo[t/]thiazol-2-yl)benzamide was made in a manner similar to Example 91. LCMS: MS (ESI) [M+H]+requires m/z 669.14, found m/z 669.2. 'HNMR (400 MHz, METHANOL-dj 5 ppm 8.21 (s, 1 H), 8.08 (br s, 1 H), 7.79 (s, 1 H), 7.49 (br s, 1 H), 5.00 (s, 2 H), 4.47 (s, 2 H), 3.75 (dt, J = 8.71, 5.13 Hz, 6 H), 3.42 - 3.45 (m, 4 H), 3.41 (s, 6 H), 3.35 (t, J = 5.29 Hz, 2 H), 2.97 (s, 3 H).
Example 94
5-chloro-2-hydroxy-3-methyl-jV-(4-(l-methylpiperidin-4-yl)-6- (trifluoromethy l)b enzo[J] th i azol -2-y l)b enzami de (94)
-261-
SUBSTITUTE SHEET ( RULE 26 )
[0575] To a solution of 2-hydroxy-3 -methylbenzoic acid (3.04 g, 20.0 mmol) in DMF (50 mL) was added NCS (2.67 g, 20.0 mmol), and the mixture was stirred at room temperature overnight. After completion of the reaction as indicated by LCMS, the reaction mixture was poured into H2O (150 mL), extracted with ethyl acetate (200 mLx 4) and concentrated under reduced pressure to give a residue, which was purified by prep-HPLC to give 5-chloro-2-hydroxy-3-methylbenzoic acid (704.2 mg, 19% yield) as an off-white solid. MS Calcd.: 186.0; MS Found: 187.1 [M+H] +.
[0576J To a solution of 2-bromo-4-(trifluoromethyl) aniline (23.9 g, 100.0 mmol) and NH4SCN (15.2 g, 200.0 mmol) in CHCh (500 mL) was added TFA (17.1 g, 150.0 mmol), and the mixture was stirred at 70 °C overnight. After completion of the reaction as indicated by LCMS, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (eluted with 5% MeOH/DCM) to afford l-(2-bromo-4-(trifluoromethyl)phenyl)thiourea (22.5 g, 76% yield) as a yellow solid. MS Calcd.: 297.9; MS Found: 299.0 [M+H] +.
-262-
SUBSTITUTE SHEET ( RULE 26 ) [0577] To a solution of l-(2-bromo-4-(trifluoromethyl) phenyl) thiourea (14.9 g, 50.0 mmol) in CHCh (300 mL) was added dropwise Br (15.8 g, 100.0 mmol) at 0 °C, and then the mixture was stirred at 70 °C overnight. After completion of the reaction as indicated by LCMS, the reaction mixture was filtered. The solid was dissolved in water, adjusted to pH 10 with 2N NaOH (aq), and extracted with ethyl acetate. The combined organic extract was washed with brine, dried over Na2SO4 and concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (eluted with 5% MeOH/DCM) to give 4-bromo-6-(trifluoromethyl)benzo[t/]thiazol-2-amine (6.0 g, 40% yield) as a yellow solid. MS Calcd.: 295.9; MS Found: 297.0 [M+H] +.
[0578| To a solution of 4-bromo-6-(trifluoromethyl)benzo[d]thiazol-2-amine (592.0 mg, 2.0 mmol), l-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,2,3,6- tetrahydropyridine (669.0 mg, 3.0 mmol) and K3PO4 (1.27 g, 6.0 mmol) in 1,4-dioxane (10 mL) and H2O (1 mL) was added 1, 1'-Bis (di-t-butylphosphino) ferrocene palladium dichloride (118.4 mg, 20% w/w), and then the mixture was stirred at 80 °C in a microwave reactor for 1 hour. After completion of the reaction as indicated by LCMS, the reaction mixture concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (eluted with 5% MeOH/DCM) to give 4-(l-methyl-l, 2,3,6- tetrahydropyridin-4-yl)-6-(trifluoromethyl)benzo[c/]thiazol-2-amine (412.5 mg, 65% yield) as yellow solid. MS Calcd.: 313.1; MS Found: 314.2 [M+H] +.
[0579] To a solution of 4-(l-methyl-l,2,3,6-tetrahydropyridin-4-yl)-6- (trifluoromethyl)benzo[ ]thiazol-2-amine (412.5 mg, 1.3 mmol) in EtOH (40 mL) was added Rh(PPh3)3Cl (412.5 mg), and the mixture was stirred at 90 °C under H2 (g) at atmospheric pressure overnight. After completion of the reaction as indicated by LCMS, the reaction mixture was filtered through Celite and the filtrate was concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (eluted with 10% MeOH/DCM) to give 4-(l-methylpiperidin-4-yl)-6-(trifluoromethyl)benzo[tZ]thiazol-2-amine (312.5 mg, 75% yield) as a light yellow solid. MS Calcd.: 315.1; MS Found: 316.0 [M+H] +.
[0580] To a solution of 4-(l-methylpiperidin-4-yl)-6- (trifluoromethyl)benzo[d]thiazol-2-amine (312.5 mg, 0.99 mmol), 5-chl oro-2 -hydroxy-3- methylbenzoic acid (202.5 mg, 1.09 mmol) and E+N (303.0 mg, 33.0 mmol) in DCM (5 mL) was added EDCI (382.2 mg, 2.0 mmol) and HOBT (270.4 mg, 2.0 mmol), and the mixture was stirred at room temperature overnight. After completion of the reaction as indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (eluted with 10% MeOH/DCM),
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SUBSTITUTE SHEET ( RULE 26 ) followed by prep-HPLC to give 5-chloro-2-hydroxy-3-methyl-jV-(4-(l-methylpiperidin-4-yl)- 6-(trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (28.86 mg, 6% yield) as a white solid.
MS Calcd.: 483.1; MS Found: 484.0 [M+H] +. 'H NMR (400 MHz, DMSO-d6) 5 17.05 (d, J = 2.8 Hz, 1H), 9.27 (dd, J= 8.8, 12.0 Hz, 1H), 8.12 (s, 1H), 7.62 (d, J= 2.8 Hz, 1H), 7.32 (d, J= 12.0 Hz, 1H), 7.18 (d, J= 2.8 Hz, 1H), 3.69-3.67 (m, 1H), 3.65-3.61 (m, 2H), 3.21 (s, 2H), 2.33 (s, 3H), 2.15-2 07 (m, 4H).
Example 95
5-chloro-2-hydroxy-3-methyl-jV-(4-(morpholinomethyl)-6-(trifluoro-methyl)benzo[<i]thiazol- 2-yl)benzamide (95)
[0581] To a solution of 4-bromo-6-(trifluoromethyl)benzo[tZ]thiazol-2-amine
(5.92 g, 20.0 mmol, Example 94), triethylamine (2.1 g, 21.0 mmol), DMAP (424.2 mg, 2.0
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SUBSTITUTE SHEET ( RULE 26 ) mmol) in DCM (200 mL) was added (BochO (4.6 g, 21.0 mmol), and the mixture was stirred at room temperature overnight. After completion of the reaction as indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography (eluted with 10% ethy; acetate/petroleum ether) to give tert-butyl (4-bromo-6-(trifluoromethyl)benzo[ ] thiazol-2-yl)carbamate (6.5 g, 82% yield) as a light-yellow solid. MS Calcd.: 396.0, MS Found: 341 [M-56+H] +.
[0582] To a solution of tert-butyl (4-bromo-6-(trifluoromethyl)benzo[<7] thiazol-2- yl)carbamate (3.96 g, 10.0 mmol) in THF (50 mL) at -78 °C was added z?-BuLi (2.5 N in hexane, 8.0 mL, 20.0 mmol), and the mixture was stirred at -78 °C for 1 hour. To this mixture was added DMF (1.48 g, 20.0 mmol), and the resulting mixture was stirred at -78 °C for another 1 hour. After completion of the reaction as indicated by LCMS, the reaction mixture was poured into sat. aq. NH4CI (100 mL), extracted with ethyl acetate (100 mL x 4) and concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (eluted with 20% ethyl acetate/petroleum ether) to give tert-butyl (4- formyl-6-(trifluorom ethyl )benzo[ ]thiazol-2-yl)carbamate (2.03 g, 58% yield) as a lightyellow solid. MS Calcd.: 346.1; MS Found: 291.0 [M-56+H] +.
|0583] To a solution of tert-butyl (4-formyl-6-(trifluoromcthyl)bcnzo[t/Jthiazol-2- yl)carbamate (346.0 mg, 1.0 mmol) and morpholine (435.0 mg, 5.0 mmol) in CH3OH (10 mL) was added NaBFFCN (126 mg, 2.0 mmol). The mixture was stirred at room temperature overnight. After completion of the reaction as indicated by LCMS, the reaction mixture was poured into water, extracted with ethyl acetate (30 mL x 4) and concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (eluted with 33% ethyl acetate/petroleum ether) to give Zc/7-butyl (4-(morpholinomethyl)-6- (trifluoromethyl)benzo[t/]thiazol-2-yl)carbamate (226.3 mg, 54% yield) as a light-yellow solid. MS Calcd.: 417.1; MS Found: 418.0 [M+H]+.
|0584] To a solution of ZcrZ-butyl (4-(morpholinomethyl)-6- (trifluoromethyl)benzo[t/]thiazol-2-yl)carbamate (226.3 mg, 0.54 mmol) in DCM (8 mL) was added TFA (2 mL) and the resulting solution was stirred at room temperature overnight. After completion of the reaction as indicated by LCMS, the reaction mixture was concentrated, poured into 0.05 NNaOH aq. (20 mL), extracted with DCM (30 mL x 4) and concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (eluted with 5% MeOH/DCM) to give 4-(morpholinomethyl)-6- (trifluoromethyl)benzo[J]thiazol-2-amine (143.6 mg, 83% yield) as a light-yellow solid. MS Calcd.: 317.1; MS Found: 318.0 [M+H] +.
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SUBSTITUTE SHEET ( RULE 26 ) [05851 To a solution of 4-(morphol i nomethyl )-6-(trifluoromethyl)benzo[t/]thiazol-
2-amine (143.6 mg, 0.45 mmol), 5-chloro-2-hydroxy-3-methylbenzoic acid (92.7 mg, 0.50 mmol) and F.hN (136.4 mg, 1.35 mmol) in DCM (5 mb) was added EDCI (171.9 mg, 0 90 mmol) and HOBT (121.5 mg, 0.90 mmol), and the mixture was stirred at room temperature overnight. After completion of the reaction as indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (eluted with 10% MeOH/DCM), followed by prep-HPLC to give 5- chloro-2-hydroxy-3-methyl-A-(4-(morpholinomethyl)-6-(trifluoromethyl)benzo[t/]thiazol-2- yl)benzmide (70.8 mg, 33% yield) as a light-yellow solid. MS Calcd.: 485.1; MS Found: 486.1 [M+H] +. XH NMR (400 MHz, DMSO-d6) 5 8.36 (s, 1H), 7.80 (s, 1H), 7.74 (s, 1H), 7.26 (d, J= 2.0 Hz, 1H), 4.63 (s, 2H), 3.78 (s, 4H), 3.18 (d, J = 7.2 Hz, 4H), 2.18 (s, 3H).
Example 96
5-chloro-A-(4-((dimethylamino)rnethyl)-6-(trifluoromethyl)benzo[<7]thiazol-2-yl)-2-hydroxy- 3 -methylbenzamide (96)
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SUBSTITUTE SHEET ( RULE 26 ) [0586| To a solution of tert-butyl (4-formyl-6-(trifluoromethyl)benzo[z/]thiazol-2- yl)carbamate (346.0 mg, 1.0 mmol, Example 95) and dimethylamine (2.5 mL, 2 N in THF) in CH3OH (10 mL) was added NaBH CN (126 mg, 2.0 mmol), and the mixture was stirred at room temperature overnight. After completion of the reaction indicated by LCMS, the reaction mixture was poured on water, extracted with ethyl acetate (30 mL x 4) and concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (eluted with 33% ethyl acetate/petroleum ether) to give tert-butyl (4- ((dimethylamino)methyl)-6-(trifluoromethyl)benzo[ ]thiazol-2- yl)carbamate (213.5 mg, 57% yield) as a light-yellow solid. MS Calcd.: 375.1, MS Found: 376 0 [M+H] +
[0587| To a solution of tert-butyl (4-((dimethylamino)methyl)-6- (trifluoromethyl)benzo[ ]thiazol-2- yl)carbamate (213.5 mg, 0.57 mmol) in DCM (8 mL) was added TFA (2 mL). The resulting solution was stirred at room temperature overnight. After completion of the reaction as indicated by LCMS, the reaction mixture was concentrated, poured into 0.05 NNaOH aq. (20 mL), extracted with DCM (30 mL x 4) and concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (eluted with 5% MeOH/DCM) to give 4-((dimethylamino)methyl)- 6-(trifluoromethyl)benzo[d]thiazol-2-amine (140.2 mg, 89% yield) as a light-yellow solid. MS Calcd.: 275.1; MS Found: 276.1 [M+H] +.
[0588| To a solution of 4-((dimethylamino)methyl)-6- (trifluoromethyl)benzo[d]thiazol-2-amine (123.8 mg, 0.45 mmol), 5-chl oro-2 -hydroxy-3- methylbenzoic acid (92.7 mg, 0.50 mmol) and EtsN (136.4 mg, 1.35 mmol) in DCM (5 mL) was added EDCI (171.9 mg, 0.90 mmol) and HOBT (121.5 mg, 0.90 mmol). The resulting mixture was stirred at room temperature overnight. After completion of the reaction as indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (eluted with 10% MeOH/DCM), followed by prep-HPLC to give 5-chloro-A-(4-((dimethylamino)methyl)-6- (trifluoromethyl)benzo[c/]thiazol-2-yl)-2-hydroxy-3-methylbenzamide (68.4 mg, 34% yield) as an off-white solid. MS Calcd.: 443.1; MS Found: 444.1 [M+H] +. 'H NMR (400 MHz, DMSO-dr,) 6 16.44 (s, 1H), 9.53 (br, 1H), 8.33 (s, 1H), 7.78 (s, 1H), 7.70 (d, 7= 2.4 Hz, 1H), 7.22 (d, J= 2.4 Hz, 1H), 4.69 (s, 2H), 2.03 (s, 6H), 2.17 (s, 3H).
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SUBSTITUTE SHEET ( RULE 26 ) Example 97
5-chloro-2-hydroxy-3-m ethoxy -/V-(6-(trifluoromethyl)benzo[</]thiazol-2-yl)benzamide (97)
[0589] To a solution of 2-hydroxy-3 -methoxybenzoic acid (200 mg, 1.19 mmol) in dioxane (2 mL) was added HC1 (6 M, 208.15 pL). Then H2O2 (148 mg, 1.31 mmol, 125.72 pL, 30% purity) was added with rapid stirring of 1 minute, and then the mixture was stirred at 20 °C for 12 h. The reaction mixture was quenched by addition 10% Na2SOs (5 mL), concentrated under reduced pressure to remove solvent, diluted with water (5 mL) and adjusted to pH 9 with 2 N NaOH. then extracted with ethyl acetate (5 mL x 3). The clear solution was acidified with IN HC1 to pH 5 and then extracted with ethyl acetate (10 mL x 3). The combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give 5-chloro-2-hydroxy-3-methoxybenzoic acid (140 mg, crude) as a yellow solid. MS (ESI) [M-H]' requires m/z 201.00, found m/z 200.9
[0590] A mixture of 5-chloro-2-hydroxy-3-methoxybenzoic acid (100 mg, 493.60 pmol), 6-(trifluoromethyl)benzo[tZ]thiazol-2-amine (107 mg, 493.60 pmol), EDCI (94 mg, 493.60 pmol), and HOBt (66 mg, 493.60 pmol) in DCM (3 mL), was stirred at 50 °C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 5-chloro-2-hydroxy-3- methoxy-JV-(6-(trifluoromethyl)benzo[</]thiazol-2-yl)benzamide (59.49 mg, 100% purity) as a white solid. 'H NMR (400 MHz, DMSO-tL) d ppm 8.54 (s, 1 H), 7.90 (br d, J= 7.94 Hz, 1 H), 7.79 (br d, J= 8.60 Hz, 1 H), 7.52 (d, J= 2.43 Hz, 1 H), 7.28 (s, 1 H), 3.90 (s, 3 H). MS (ESI) [M-H]' requires m/z 403.01, found m/z 402.9
Example 98
5-chloro-2-hydroxy-3-(tetrahydrofuran-3-yl)-7V-(6-(trifluoromethyl)pyridin-3-yl)benzamide (98)
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SUBSTITUTE SHEET ( RULE 26 )
[0591] To a stirred solution of methyl 5-chloro-2-methoxy-3-(tetrahydrofuran-3- yl)benzoate (929 mg, 3.44 mmol, Example 14) in MeOH (10 mL) was added 10.0 mL IN KOH solution. The resulting mixture was stirred at rt overnight. After completion of the reaction, the mixture was partitioned between IN HC1 and ethyl acetate. The organic layer was dried over sodium sulfate and concentrated under reduced pressure to afford a residue of 5-chloro-2-methoxy-3-(tetrahydrofuran-3-yl)benzoic acid. The residue was used directly in the next step.
[0592] 5-chloro-2-methoxy-3-(tetrahydrofuran-3-yl)benzoic acid (81 mg, 0.316 mmol) was dissolved in DCM (5.0 mL), followed by the addition of a catalytic amount of DMF (1 drop) and oxalyl chloride (33 pL, 0. 379 mmol) respectively. The reaction was allowed to stir at rt for 30 minutes then concentrated in vacuo. The residue was re-dissolved in THF (5.0 mL), and Hunig’s base (66 pL, 0.379 mmol) and 6-(trifluorom ethyl)pyri din-3 - amine (51 mg, 0.316 mmol) were added. The mixture was stirred at rt for 48 hours before silica gel was added to quench the reaction. Solvent was evaporated and the resulting residue was purified via silica gel column chromatography to yield 5-chloro-2-methoxy-3- (tetrahydrofuran-3-yl)- V-(6-(trifluoromethyl)pyridin-3-yl)benzamide (124 mg, 97%) as a yellow solid. MS (ESI) [M+H]+ requires m/z 401.0, found m/z 401.1.
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SUBSTITUTE SHEET ( RULE 26 ) [0593| A solution of 5-chloro-2-methoxy-3-(tetrahydrofuran-3-yl)-A-(6- (trifluoromethyl)pyridin-3-yl)benzamide (124 mg, 0.31 mmol) in DMF (5 mL) was mixed with sodium ethanethiolate (130 mg, 1.55 mmol) and the resulting suspension was heated at 130 °C overnight. After completion of the reaction, IN HC1 was added to the reaction and extracted with ethyl acetate two times. The combined ethyl acetate layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel column chromatography to give 5-chloro-2-hydroxy-3-(tetrahydrofuran-3-yl)-A-(6- (trifluoromethyl)pyridin-3-yl)benzamide (103 mg, 87%) as a yellow solid. XHNMR (300 MHz, Chloroform-cZ) 5 11.73 (s, 1H), 9.09 (s, 1H), 8.84 (s, 1H), 8.47 (d, J= 8.7 Hz, 1H), 7 84 - 7.57 (m, 2H), 7.36 (d, J= 2.4 Hz, 1H), 4.24 - 4.04 (m, 2H), 4.01 - 3.83 (m, 2H), 3.84 - 3.71 (m, 1H), 2.53 - 2.35 (m, 1H), 2.14 - 1.96 (m, 1H). MS (ESI) [M-H]’ requires m/z 385.1, found m/z 385.3.
Example 100
5-chloro-2-hydroxy-A-(4-(((4-hydroxybenzyl)oxy)methyl)-6- (trifluoromethyl)benzo[6f]thiazol-2-yl)-3 -methylbenzamide (100)
[0594] To a solution of Zc/7-butyl (4-(bromomethyl)-6- (trifluoromethyl)benzo[d]thiazol-2-yl)(tert-butoxycarbonyl)carbamate (546 mg, 1.07 mmol,
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SUBSTITUTE SHEET ( RULE 26 ) Example 64) and (4-bromophenyl)methanol (200 mg, 1.07 mmol) in THF (10 mL) was added dropwise t-BuOK (1 M, 4.2 mL) at 0°C under Nz. The resulting mixture was stirred at 0°C for 1 hr. The reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (5 mL x 3). The combined organic layer was washed with brine (10 mL x 2), dried over NazSC , filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®, 12 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% ethyl acetate/petroleum ether gradient @70 mL/min) to afford compound tert-butyl (4-(((4-bromobenzyl)oxy)methyl)-6-(trifluoromethyl)benzo[ ]thiazol-2- yl)(tert-butoxycarbonyl)carbamate (0.2 g, 168.43 pmol, 15.75% yield, 52% purity) as yellow oil.
10595] tert-butyl (4-(((4-bromobenzyl)oxy)methyl)-6- (trifluoromethyl)benzo[d]thiazol-2-yl)(tert-butoxycarbonyl)carbamate (0.2 g, 323.90 pmol), KOH (363 mg, 6.48 mmol), Pd2(dba)3 (5 mg, 6.48 pmol)and di-tert-butyl-[2-(2,4,6- triisopropylphenyl)phenyl]phosphane (11 mg, 25.91 pmol) in dioxane (5 mL) and HzO (5 mL) was degassed and purged with N2 3 times, and then the mixture was stirred at 110°C for 2 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with water (5 mL) and extracted with ethyl acetate (5 mL x 3). The combined organic layer was washed with brine (3 mL x 3); dried over Na?SC>4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiOz, petroleum ether/ethyl acetate 3/1) to afford compound tert-butyl (tert-butoxycarbonyl)(4-(((4-hydroxybenzyl)oxy)methyl)-6-(trifluoromethyl)benzo[</]thiazol- 2-yl)carbamate (0.1 g, 100.98 pmol, 31.18% yield, 56% purity) as yellow oil.
[0596] A solution tert-butyl (tert-butoxycarbonyl)(4-(((4- hydroxybenzyl)oxy)methyl)-6-(trifluoromethyl)benzo[</]thiazol-2-yl)carbamate (0.07 g, 126.22 pmol) in HFIP (3 mL) was stirred at 80°C for 1.5 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep- TLC (S1O2, petroleum ether/ethyl acetate 1/1) to afford 4-(((2-amino-6- (trifluoromethyl)benzo[r/]thiazol-4-yl)methoxy)methyl)phenol (17 mg, 26.72 pmol, 21.17% yield, 55.7% purity) as yellow oil.
[0597] A mixture of 4-(((2-amino-6-(trifluoromethyl)benzo[</]thiazol-4- yl)methoxy)methyl)phenol (25 mg, 70.55 pmol), 5-chloro-2-hydroxy-3-methylbenzoic acid (15 mg, 84.66 pmol) in DCM (3 mL) was added EDCI (16 mg, 84.66 pmol) and HOBt (14 mg, 105.83 pmol), and then the mixture was stirred at 45°C for 12 hrs. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by
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SUBSTITUTE SHEET ( RULE 26 ) prep-HPLC (neutral condition) to afford 5-chloro-2-hydroxy-JV-(4-(((4- hydroxybenzyl)oxy)methyl)-6-(trifluoromethyl)benzo[</]thiazol-2-yl)-3-methylbenzamide (7.54 mg, 14 31 pmol, 20.29% yield, 99.27% purity) as a white solid. LH NMR (400 MHz, DMSO-Jr,) 8 ppm 9.41 (br s, 1 H), 8.44 (s, 1 H), 8.02 (br s, 1 H), 7.75 (s, 1 H), 7.47 (br s, 1 H) 7.19 (d, J= 8.38 Hz, 2 H), 6.74 (d, J= 8.38 Hz, 2 H), 4.93 (s, 2 H), 4.55 (s, 2 H), 2.16 - 2.27 (m, 3 H). MS (ESI) [M+H]+ requires m/z 523.1, found m/z 522.9
Example 101
5-chloro-2-hydroxy-.V-(4-((2-methoxyacetamido)methyl )-6-(trifluoromethyl)benzo[6/]thiazol-
2-yl)-3-methylbenzamide (101)
10598] To a solution of Zc/7-butyl (4-(bromomethyl)-6- (trifluoromethyl)benzo[rZ]thiazol-2-yl)(tert-butoxycarbonyl)carbamate (500 mg, 977.8 pmol, Example 64) in DMF (5 mL) was added potassium l,3-dioxoisoindolin-2-ide (271 mg, 1.47 mmol). The mixture was stirred at 25°C for 12 hrs. The reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (5 mL x 3). The combined organic layers were washed with brine (15 mL x 2), dried over Na2SC>4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% ethyl
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SUBSTITUTE SHEET ( RULE 26 ) acetate /petroleum ether gradient @ 70 mL/min) to afford Zc/7-butyl (tert-butoxycarbonyl)(4- (h 1 ,3-di oxoisoindolin-2-yl )methyl)-6-ftrifluorom ethyl )benzo[<7]thiazol-2-yl)carbamate (320 mg, 554.0 pmol, 56.6% yield) as a white solid.
|0599] To a suspension of tert-butyl (tert-butoxycarbonyl)(4-((l,3- dioxoi soindolin-2-yl )methyl)-6-(trifluoromethyl)benzo[c/]thiazol -2-yl )carbam ate (320 mg, 554 04 pmol) in EtOH (5 mb) was added N2H4.H2O (56 mg, 1.1 mmol, 54 95 uL, 98% purity). The mixture was stirred at 25°C for 5 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with DCM (20 mL), then fdtered and concentrated under reduced pressure to afford tert-butyl (4-(aminomethyl)-6- (trifluoromethyl)benzo[J]thiazol-2-yl)(tert-butoxycarbonyl)carbamate (150 mg, crude) as a white solid.
|0600] To a solution of t terZ-butyl (4-(aminomethyl)-6- (trifluoromethyl)benzo[J]thiazol-2-yl)(tert-butoxycarbonyl)carbamate (130 mg, 374.26 pmol) in DCM (3 mL) was added TEA (56 mg, 561.38 umol, 78 pL), then 2-methoxyacetyl chloride (48 mg, 449.11 pmol, 40 pL) was added at 0°C. The resulting mixture was stirred at 20°C for 12 hrs. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-TLC (SiCb, petroleum ether/ethyl acetate 1/1) to afford tert-butyl (tert-butoxycarbonyl)(4-((2-methoxyacetamido)methyl)-6- (trifluoromethyl)benzo[t/]thiazol-2-yl)carbamate (140 mg, 298.41 pmol, 79.74% yield, 89.4% purity) as yellow oil.
|0601] To a solution of tert-butyl (tert-butoxycarbonyl)(4-((2- methoxyacetamido)methyl)-6-(trifluoromethyl)benzo[d]thiazol-2-yl)carbamate (140 mg, 333.80 pmol) in DCM (3 mL) was added TFA (1 mL). The mixture was stirred at 25°C for 26 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with sat NaHCCh (20 mL) and extracted with ethyl acetate (3 mL x 3). The combined organic layer was washed with brine (10 mL x 2), dried over JSfeSCL, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep- TLC ( Si O2, petroleum ether/ethyl acetate 1/1) to afford compound A-((2-amino-6- (trifluoromethyl)benzo[d]thiazol-4-yl)methyl)-2-methoxyacetamide (0.1 g, 295.77 pmol, 88.61% yield, 94.44% purity) as a white solid.
|0602] To a solution of A-((2-amino-6-(trifluoromethyl)benzo[d]thiazol-4- yl)methyl)-2-methoxyacetamide (0.08 g, 250.55 umol), 5 -chi oro-2-hydroxy-3 -methylbenzoic acid (46 mg, 250.55 pmol) in DCM (3 mL) was added EDCI (57 mg, 300.66 pmol) and HOBt (50 mg, 375.82 pmol). The mixture was stirred at 50°C for 12 hrs. The reaction
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SUBSTITUTE SHEET ( RULE 26 ) mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 5-chloro-2-hydroxy-A'-(4-(('2- methoxyacetamido)methyl)-6-(trifluoromethyl)benzo[<7]thiazol-2-yl)-3-methylbenzamide (21.24 mg, 43.54 pmol, 17.38% yield, 100% purity) as a white solid. 1H NMR (400 MHz, DMSO-d6) S ppm 8.53 - 8.60 (m, 1 H), 8.41 (s, 1 H), 8.02 (br s, 1 H), 7.56 (br s, 1 H), 7.47 (br s, 1 H), 4 74 (br s, 2 H), 3.92 (s, 2 H), 3.34 (s, 3 H), 2.21 (s, 3 H) MS (ESI) [M+H]+ requires m/z 488.1 , found m/z 487.9
Example 102
5-chloro-3-((cA-2,6-dimethylmorpholino)methyl)-2-hydroxy-A-(4-(methoxymethyl)-6-
(trifluoromethyl)benzo[d]thiazol-2-yl)benzamide (102)
[0603| A solution of /evV-butyl (4-(bromomethyl)-6- (trifluoromethyl)benzo[t/]thiazol-2-yl)(tert-butoxycarbonyl)carbamate (0.4 g, 782.24 pmol. Example 64) in NaOMe (10 mL, 30% in MeOH) was stirred at 30°C for 5 h. The reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (5 mL * 3). The combined organic layer was washed with brine (30 mL), dried over NazSCh, filtered and
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SUBSTITUTE SHEET ( RULE 26 ) concentrated under reduced pressure to afford /cz'Z-butyl (4-(methoxymethyl)-6- (trifluoromethyl)benzo[d]thiazol-2-yl)carbamate ( 0.25 g, crude) as a yellow oil.
|0604] To a solution of tert-butyl (4-(methoxymethyl)-6- (trifluoromethyl)benzo[z/]thiazol-2-yl)carbamate (0.25 g, 689.91 pmol) in DCM (3 mL) was added TFA (1 mL). The mixture was stirred at 20°C for 1 hr. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with sat NaHCCh (10 mL) and extracted with ethyl acetate (5 mL x 3). The combined organic layer was washed with brine (15 mL x 3), dried over NazSCL, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/ethyl acetate 1/1) to afford 4-(methoxymethyl)-6-(trifluoromethyl)benzo[d]thiazol-2- amine (136 mg, 475.03 pmol, 68.85% yield, 91.6% purity) as a yellow solid.
|0605] To a solution of 4-(methoxymethyl)-6-(trifluoromethyl)benzo[z/]thiazol-2- amine (60 mg, 228.79 pmol), 5-chloro-3-[(cis,2,6-dimethylmorpholin-4-yl)methyl]-2- hydroxy-benzoic acid (82 mg, 274.55 pmol) in DCM (4 mL) was added EDCI (52 mg, 274.55 pmol) and HOBt (46 mg, 343 pmol). The mixture was stirred at 45°C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (neutral condition) to afford compound 5-chloro-3-((czs-2,6- dimethylmorpholino)methyl)-2 -hydroxy -/V-(4-(methoxymethyl)-6- (trifluoromethyl)benzo[z/]thiazol-2-yl)benzamide (23.67 mg, 42.37 pmol, 18.52% yield, 97.373% purity) as a white solid. 'H NMR (400 MHz, DMSO-zL) 6 ppm 8.35 (s, 1 H), 7.81 (d, J= 2.87 Hz, 1 H), 7.64 (s, 1 H), 7.38 (d, J= 3.09 Hz, 1 H), 4.84 (s, 2 H), 4.22 (br s, 2 H), 3.91 (br dd, J= 931, 6.28 Hz, 2 H), 3.42 (s, 3 H), 3.29 - 3.36 (m, 4 H), 1.13 (d, J = 6.17 Hz, 6 H). MS (ESI) [M+H]+ requires m/z 544.1 , found m/z 544.1
Example 103
5-chloro-3-((czs-2,6-dimethylmorpholino)methyl)-/V-(4-(ethoxymethyl)-6- (trifluoromethyl)benzo[z/]thiazol-2-yl)-2-hydroxybenzamide (103)
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SUBSTITUTE SHEET ( RULE 26 ) [0606J 5-chloro-3-((czs-2,6-dimethylmorpholino)methyl)-jV-(4-(ethoxymethyl)-6- (trifluoromethyl)benzo[c/]thiazol-2-yl )-2 -hydroxybenzamide was prepared in a manner similar to Example 102.
[0607! LCMS: MS (ESI) [M+H]+ requires m/z 558.1, found m/z 558.1XH NMR (400 MHz, DMSO-d6) 5 ppm 8.37 (s, 1 H), 7.90 (br s, 1 H), 7.68 (s, 1 H), 7.48 (br s, 1 H), 4.89 (s, 2 H), 4.26 (s, 2 H), 3.91 (br dd, J= 10.03, 6.06 Hz, 2 H), 3.62 (q, J= 6.98 Hz, 2 H), 3.34 (br d, J= 12.13 Hz, 2 H), 2.69 (br t, J= 11.36 Hz, 2 H), 1.21 (t, J= 7.06 Hz, 3 H), 1.13 (d, J = 6.39 Hz, 6 H
Example 104
5-chloro-3-((czs-2,6-dimethylmorpholino)methyl)-JV-(4-((2-methoxy ethoxy )methyl)-6- (trifluoromethyl)benzo|X]thiazol-2-yl)-2-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6- (hydroxymethyl )tetrahydro-2/7-pyran-2-yl)oxy)benzamide (104)
[0608| 5 -chloro-3-((czs-2,6-dimethylmorpholino)methyl)-2-hydroxy-JV-(4-((2- methoxyethoxy)methyl)-6-(trifluoromethyl)benzo[z/]thiazol-2-yl)benzamide (100 mg, 170.06
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SUBSTITUTE SHEET ( RULE 26 ) pmol, Example 64), (2/?,3/<4A’,5/?,6A’)-2-(acetoxymethyl )-6-bromotetrahydro-2//-pyran-
3.4.5-triyl triacetate (139 mg, 340.11 pmol), AgzO (78 mg, 340.11 pmol) in ACN (9 mL), mixed then stirred at 30°C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-TLC (SiCh, petroleum ether/ethyl acetate 5/7) to afford (2A,3A,4S,5A,65)-2-(acetoxymethyl)-6-(4-chloro-2-((c/s-2,6- dimethylmorpholino)methyl)-6-((4-((2 -methoxy ethoxy)methyl)-6-
(tri fl uoromethyl)benzo[<7]thiazol -2-yl (carbamoyl (phenoxy (tetrahydro-2//-pyran-3, 4, 5-triyl triacetate (85 mg, 72.20 pmol, 42.45% yield, 78% purity) as yellow oil.
[0609] To (27?,3A,41S,57?,61S)-2-(acetoxymethyl)-6-(4-chloro-2-((cA-2,6- dimethylmorpholino)methyl)-6-((4-((2-methoxyethoxy)methyl)-6-
(tri fl uoromcthyl)bcnzo[t/]thiazol -2-yl (carbamoyl (phenoxy )tctrahydro-27/-pyran-3, 4, 5-triyl triacetate (85 mg, 92.56 pmol) in MeOH (6 mL) was added NaOMe (33 mg, 185.12 pmol, 30% purity). The mixture was stirred at 25°C for 5 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (neutral condition) to afford 5-chloro-3-((cA-2,6-dimethylmorpholino)methyl)-A-(4-((2- methoxy ethoxy )methyl)-6-(trifluoromethyl)benzo[<7]thiazol-2-yl)-2-(((2S,3R,4S, 5S,6R)-
3.4.5-trihydroxy-6-(hydroxymcthyl)tctrahydro-27/-pyran-2-yl)oxy (benzamide (3.72 mg, 4.82 pmol, 5.21% yield, 97.284% purity) as a white solid. ’H NMR (400 MHz, DMSO-r/y) 5 ppm 12.74 (br s, 1 H), 8.44 (s, 1 H), 7.72 (s, 1 H), 7.61 (s, 1 H), 7.56 (d, J= 1.98 Hz, 1 H), 5.42 (br s, 1 H), 4.94 (s, 2 H), 4.77 (d, J = 5.29 Hz, 1 H), 4.62 (d, J= 7.72 Hz, 1 H), 4.11 - 4.20 (m, 2 H), 3.65 - 3.77 (m, 3 H), 3.47 - 3.60 (m, 7 H), 3.25 (s, 3 H), 2.83 (br d, J= 9.70 Hz, 1 H), 2.65 - 2.69 (m, 1 H), 1.65 - 1.82 (m, 2 H), 1.03 (br t, J= 7.06 Hz, 6 H). MS (ESI) [M+H]+ requires m/z 750.2 , found m/z 750.0
Example 105
5-chloro-3-((/ra«5-2,6-dimethylmorpholino)methyl)-2-hydroxy-A-(4-((2- m cthoxycthoxy (methyl )-6-(trifluoromcthyl)bcnzo[t/]thi azol -2-yl (benzamide (105)
SUBSTITUTE SHEET ( RULE 26 )
[0610| To a solution of 5-chloro-3-formyl-2-hydroxybenzoic acid (0.4 g, 1.99 mmol) and trans-2,6-dimethylmorpholine (459 mg, 3.99 mmol, 491 gL) in THF (30 mL) was added acetic acid (119 mg, 1.99 mmol, 114 gL) until the pH reached ~ 5 - 6. The mixture was stirred at 30°C for 3 h then NaBH(OAc)3 (2.5 g, 11.97 mmol) was added. The mixture was stirred at 30°C for another 3 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (neutral condition) to afford 5-chloro-3-((/ra/AS-2,6-dimethylmorpholino)methyl)-2-hydroxybenzoic acid (250 mg, 748.96 gmol, 37.56% yield, 89.8% purity) as a white solid.
[0611j To a solution of fert-butyl (4-(bromomethyl)-6- (trifluoromethyl)benzo[c/]thiazol-2-yl)(/c/7-butoxycarbonyl)carbarnate (400 mg, 586.68 gmol), 2-methoxyethanol (89 mg, 1.17 mmol, 92 gL) in THF (10 mL) was added Z-BuOK (IM, 2 mL) at 0°C under N2 atmosphere. The suspension was degassed and purged with N2 3 times, then the mixture was stirred at 20°C for 12 h. The reaction mixture was diluted with
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SUBSTITUTE SHEET ( RULE 26 ) water (30 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layer was washed with brine (100 mL), dried over Na2SC>4, filtered and concentrated under reduced pressure to afford tert-butyl (4-((2-methoxyethoxy)methyl)-6-
(trifluoromethyl)benzo[<7]thi azol -2-yl (carbarn ate (230 mg, crude) as yellow oil.
|0612] To a solution of tert-butyl (4-((2-methoxyethoxy)methyl)-6- (trifluoromethyl)benzo[d]thiazol-2-yl)carbamate (230 mg, 565.92 pmol) in DCM (7 mL) was added TFA (1 mL). The mixture was stirred at 20°C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with NaHCCh (20 mL) and extracted with ethyl acetate (4 mL x 3). The combined organic layer was washed with brine (10 mL x 3), dried over NaiSCL, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/ethyl acetate 1/1) to afford 4-((2 -methoxyethoxy )methyl)-6- (trifluoromethyl)benzo[<7]thiazol-2-amine (160 mg, 411.10 pmol, 72.64% yield, 78.7% purity) as a yellow solid.
|0613] To a solution of 5-chloro-3-((teans-2,6-dimethylmorpholino)methyl)-2- hydroxybenzoic acid (70 mg, 235.06 pmol), 4-((2-methoxyethoxy)methyl)-6- (trifluoromethyl)benzo[d]thiazol-2-amine (60 mg, 195.88 pmol) in DCM (4 mL) was added EDCI (45 mg, 235.06 pmol) and HOBt (39 mg, 293.83 pmol). The mixture was stirred at 45°C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford 5-chloro-3- ((/rans-2,6-dimethylmorpholino)methyl)-2-hydroxy-7V-(4-((2-methoxy ethoxy )methyl)-6- (trifluoromethyl)benzo|7/]thiazol-2-yl)benzamide (52.88 mg, 80.76 pmol, 41.23% yield, 95.37% purity, HC1) as a white solid. H NMR (400 MHz, DMSCM,) 5 ppm 8.45 (s, 1 H), 8.07 (br s, 1 H), 7.70 - 7.81 (m, 2 H), 4.95 (s, 2 H), 4.31 (s, 2 H), 4.17 (br s, 2 H), 3.70 (dd, J = 5.62, 3.67 Hz, 2 H), 3.54 (dd, J =5.62, 3.67 Hz, 2 H), 3.20 - 3.28 (m, 5 H), 2.97 (br s, 2 H), 1.24 (br s, 6 H). MS (ESI) [M+H]+ requires m/z 588.2, found m/z 588.1
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SUBSTITUTE SHEET ( RULE 26 ) Example 106
5-chloro-2-hydroxy-A-(4-((2-methoxy ethoxy )methyl)-6-(trifluoromethyl)benzo[ ]thiazol-2- yl)-3 -(pyrrolidin- 1 -ylmethyl)benzamide ( 106)
[0614| To a solution of 5-chloro-3-formyl-2-hydroxybenzoic acid (500 mg, 2.49 mmol) and pyrrolidine (195 mg, 2.74 mmol, 228 uL) in THF (15 mL) was added acetic acid (149 mg, 2.49 mmol, 142 uL) to reach a pH of - 5-6. The mixture was stirred at 30°C for 3 h then NaBH(OAc)s (3.17 g, 14.96 mmol) was added. The mixture was stirred at 30°C for another 3 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (neutral condition) to afford 5-chloro-2- hydroxy-3-(pyrrolidin-l-ylmethyl)benzoic acid (420 mg, 1.60 mmol, 64.17% yield, 97.39% purity) as a yellow solid.
[0615] 5-chloro-2-hydroxy-3-(pyrrolidin-l-ylmethyl)benzoic acid (40.07 mg, 156.71 pmol), 4-(2-methoxyethoxymethyl)-6-(trifluoromethyl)-l,3-benzothiazol-2-amine (40 mg, 130.59 umol) in DCM (4 mL) was added EDCI (30 mg, 156.71 pmol) and HOBt (26 mg, 195.88 pmol). The mixture was stirred at 45°C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep- HPLC (HC1 condition) to afford 5-chloro-2-hydroxy-jV-(4-((2-methoxyethoxy)methyl)-6- (trifluoromethyl)benzo[< ]thiazol -2 -yl)-3 -(pyrrolidin- l-ylmethyl)benzamide (98.25% purity, HC1) 49.21 mg as a white solid. 'H NMR (400 MHz, DMSO-<76) d ppm 10.00 (br s, 1 H), 8.43 (s, 1 H), 8.06 (br s, 1 H), 7.78 (s, 1 H), 7.70 (br s, 1 H), 4.94 (s, 2 H), 4.33 (br s, 2 H),
SUBSTITUTE SHEET ( RULE 26 ) 3.67 - 3.71 (m, 2 H), 3.51 - 3.57 (m, 6 H), 3.27 (s, 3 H), 1.97 (br s, 4 H). MS (ESI) [M+H]+ requires m/z 544.1, found m/z 544.11
Example 107
5-chloro-2-hydroxy-7V-(4-(methoxymethyl)-6-(trifluoromethyl)benzo[6(]thiazol-2-yl)-3- (pyrrolidin-1 -ylmethyl)benzamide (107)
[0616] 5-chloro-2-hydroxy-jV-(4-(methoxymethyl)-6- (trifluoromethyl)benzo[d]thiazol-2-yl)-3-(pyrrolidin-l-ylmethyl)benzamide was prepared in a manner similar to Example 106. LCMS: MS (ESI) [M+H]+ requires m/z 500.1, found m/z 500. fHNMR (400 MHz, DMSO-4) <5 ppm 10.06 (br s, 1 H), 8.44 (s, 1 H), 8.06 (br s, 1 H), 7.70 - 7.75 (m, 2 H), 4.85 (s, 2 H), 4.33 (s, 2 H), 3.40 (s, 7 H), 1.97 (br s, 4 H)
Example 108
(4-chloro-2-((czs-2,6-dimethylmorpholino)methyl)-6-((4-((2-methoxyethoxy)methyl)-6-
(trifluoromethyl)benzo[<7]thiazol-2-yl)carbamoyl)phenoxy)methyl acetate (108)
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SUBSTITUTE SHEET ( RULE 26 ) [0617| To a solution of 5-chloro-3-((cA-2,6-dimethylmorpholino)methyl)-2- hydroxy-/V-(4-((2 -methoxy ethoxy)methyl)-6-(trifluoromethyl)benzo[</]thiazol-2- yl)benzamide (100 mg, 170.06 pmol, Example 64), DIPEA (65 mg, 510.17 pmol, 88 pL) in ACN (4 mL), was added bromomethyl acetate (78 mg, 510.17 pmol, 50 pL) slowly, and then the mixture was stirred at 20°C for 12 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HC1 condition) to afford (4-chloro-2-((cA-2,6-dimethylmorpholino)methyl)-6-((4-((2- methoxy ethoxy )methyl)-6-(trifluoromethyl)benzo[<7]thiazol-2-yl)carbamoyl)phenoxy)methyl acetate (10.41 mg, 14.46 pmol, 8.51% yield, 96 782% purity, HC1) as a white solid. 'H NMR (400 MHz, DMSO- s) 8 ppm 13.08 (s, 1 H), 10.86 (br s, 1 H), 8.51 (s, 1 H), 8.10 (br s, 1 H), 7.88 (br s, 1 H), 7.78 (s, 1 H), 5.55 (s, 2 H), 4.96 (s, 2 H), 4.37 (br s, 2 H), 3.97 (br s, 2 H), 3.69 (dd, J= 5.62, 3.64 Hz, 2 H), 3.52 (dd, J= 5.51, 3.75 Hz, 2 H), 3.26 (s, 3 H), 2.78 (br d, J = 8.82 Hz, 2 H), 1.86 - 1.97 (m, 3 H), 1.10 - 1.19 (m, 6 H). MS (ESI) [M+H]+ requires m/z 660.2, found m/z 660.0
Example B 1
Determining mitochondrial uncoupling activity by oxygen consumption rate (OCR) assay with Seahorse instruments
[0618] By definition, mitochondrial uncoupling is the decoupling of mitochondrial electron transport chain activity (mitochondrial oxidation) from ATP synthesis by mitochondrial ATP synthase. Technically, mitochondrial uncoupling activity is defined by the ability of a chemical compound to induce oxygen consumption of cells or isolated mitochondria in the presence of a mitochondrial ATP synthase inhibitor such as oligomycin. Accordingly, mitochondrial uncoupling activity was determined by oxygen consumption rate assay with a Seahorse XF96 or a Seahorse XF24 instrument following the vendor’s instruction, using mouse myoblast C2C12 cells grown in a medium containing 10% fetal bovine serum. OCR was measured stepwise under the following conditions: first under normal growth conditions; then. OCR was measured after oligomycin was added to reach a final concentration of 2.5 pM); then OCR was measured after the compound was further added at varying concentrations. Finally, rotenone and antimycin A were added to confirm that the measurement was mitochondrial oxygen consumption (rotenone and a are mitochondrial electron transport chain inhibitors). Figure 2 shows typical results demonstrating that the tested compounds 16 and 69, are mitochondrial uncouplers which increase OCR in the presence of oligomycin.
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SUBSTITUTE SHEET ( RULE 26 ) [0619| CocR-min the minimal concentration required for each compound to induce OCR in the presence of oligomycin, of disclosed compounds, was also determined by varying the final concentration of the compound added to the Seahorse OCR analysis experiment as described above (Figure 2, varying testing compound concentration). Table 8 summarizes the mitochondrial uncoupling activity of disclosed compounds, as represented by Corp -
Table 8
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SUBSTITUTE SHEET ( RULE 26 ) \ C OCR -min: minimal concentration increasing OCR in the presence of oligomycin. ***, <5 pM; **between 5 and 10 pM; * >10 pM.
Example B2
Unique Property: Compounds Effectively Induce OCR Without Significant Loss of Mitochondrial Membrane Potential
[0620] As proton gradient is responsible for establishing mitochondrial membrane potential (MMP), the unregulated proton transport across mitochondrial inner membrane catalyzed by mitochondrial uncouplers dissipates MMP. Prior to the present disclosure, all mitochondrial uncouplers (including DNP, herein referred to as conventional uncouplers) tested have properties that increase OCR and decreased MMP, and the concentrations for increasing OCR and dissipating MMP correlated. As such, the activity for decreasing MMP is considered a second hallmark of mitochondrial uncoupling.
[06211 Mitochondrial membrane potential assay was performed using the standard TMRE (tetramethylrhodamine ethyl ester) staining method with cultured mammalian cells, the NIH-3T3 cells, C2C12 cells, and other mammalian cell lines. Cells were seeded in 6-well plates and cultured in DMEM medium supplemented with 10% fetal bovine serum and 2 mM glutamine. Cells were allowed to grow to the logarithmic growth phase prior to experiments. The cells were treated with each individual compound at various concentrations for two hours, followed by staining with TMRE at a final concentration of 100 nM for 15 minutes. The cells were then washed once with PBS, and examined under fluorescence microscopy. Cells treated with various concentrations of niclosamide ethanolamine were used as a positive control.
[06221 In some examples, mitochondrial membrane potential was measured with DiICl(5) staining. Briefly, DiICl(5) was added to the cells to 50 nM final concentration and the cells were incubated at 37°C, 5% CO2, for 15 to 30 minutes. Cells were washed once by adding 2 m of warm phosphate-buffered saline (PBS). Mitochondrial staining was determined by fluorescence microscopy with 633 nm excitation using standard filters for far- red emission.
|0623{ As illustrated in Figure 4, the conventional mitochondrial uncoupler FCCP starts to increase OCR and reduce MMP at 3 uM at the same time; at the concentration of 9 uM, MMP is totally lost and the rapid MMP loss correlates with the increase in OCR. This correlates with what is typically seen in conventional mitochondrial uncouplers. The window between the concentration showing minimal activity Cmin-ocR and the concentration of
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SUBSTITUTE SHEET ( RULE 26 ) conventional uncoupler dissipating MMP (losing 90% of MMP, defined as CIO%TMRE) is narrow, with the ratio of Cio%TMRE/Cmin-ocR around 3 (Figure 4A and B).
|0624] Surprisingly, many of the compounds of the invention exhibited a unique feature shown in Figure 5. Compound 25 started to increase OCR at around 1.0 pM, and showed very robust OCR induction at 3.0, 6.0, and 9.0 pM (Figure 5A). However, Compound 25 does not cause observable MMP reduction under these conditions (Figure 5B).
[0625] In addition, another group of MMP-retaining uncouplers were identified, as represented by Compound 64. Unlike Compound 25, which does not lead to any observable loss in MMP, Compound 64 appears to lead to a minor loss of MMP at the highest level of mitochondrial uncoupling (highest OCR), as illustrated in Figure 6. But the MMP loss is drastically lower than that induced by a conventional uncoupler such as FCCP (Figure 4).
[0626] Using the ratio between CIO%TMRE (the concentration at which 10% MMP remains, i.e. 90% MMP is lost) and Cmin-ocR (the concentration at which minimal OCR induction is observed), we categorized the MMP-retaining uncouplers into two groups. Compound 25-like compounds are denoted as MMP -Retaining Uncoupling Compounds (CIO%TMRE / Cmin-ocR>25). The Compound 64-like compounds are denoted as MMP -Retaining Uncoupling Compounds (CIO%TMRE / Cmin-ocR between 10 to 25). The conventional uncouplers, including FCCP and DNP, have a CIO%WRE / cmin-ocR that is equal to or less than 3. The compounds are summarized in Table 6.
Example B3
New Benzamide Mitochondrial Uncouplers with Drastically Improved PK profiles
[0627] For PO pharmacokinetic studies, the compound was suspended in 0.5% CMC-Na + 1.0 % Tween 80 in water at 1.0 mg/ml, dosing at 10 mg/kg by gavage. The blood concentration of the compound was then determined at 0 (pre-dosing), 5 min, 15 min, 30 min, 1 hr, 2 hr, 4 hr, 8 hr, 24 hr time points by LC-MS/MS analysis (See Tables 1 and 2, vide supra).
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SUBSTITUTE SHEET ( RULE 26 ) Example B4
Efficacy on Type 2 Diabetes and Fatty Liver In Vivo, the Effect on Blood Glucose, Blood Insulin, and Fatty Liver in High-Fat Diet (HFD)-Induced Diabetic Mouse Model and db/db Mouse Model.
|0628] To demonstrate the efficacy of compounds of the invention in treating type 2 diabetes and steatosis in vivo, Compound 25 (Figure 11-15) and other compounds (Table 9) were tested in clinically relevant mouse models to show the effect on blood glucose, blood insulin concentration, liver steatosis, as well as other parameters such as blood lipid and cholesterol. Figure 11 shows results from db/db mice, and Figure 12-15, show results from HFD induced diabetic model. For HFD induced diabetic mouse model, C57B16 mice (male, n = 6) were induced to obese and diabetic conditions by feeding high-fat diet (HFD, 60% fat calories, Research Diet, Inc. New Brunswick, NJ) for 3-4 months. The mice were then treated with P.O. dosing (gavage) of 5 mg/kg Compound 25 or Compound 64 once daily, or vehicle, for 3 weeks. During the 3-week period, the mice continued feeding HFD. The db/db (BKS.CgDock7m +/+ Lepr db/J; stock# 000642) mice (male, n = 6) purchased from the Jackson Laboratory (Bar Harbor, ME), were treated with P.O. dosing (gavage) of 5 mg/kg Compound 25 or Compound 64 once daily, or vehicle, for 3 weeks. The blood glucose levels were measured after an overnight fast, plasma insulin levels were tested after 5-6 hours fast, and the glycated HbAlc levels were determined in whole blood samples. The results (Figure 11- Figure 13) show Compound 25 and Compound 64 are highly efficacious in reducing blood glucose, blood insulin, and glycated HbAlc in the clinically relevant models.
|0629] Compound 25 is efficacious in reducing blood triglyceride levels and non- HDL cholesterol levels induced by high-fat diet feeding (Figure 13).
[0630] The effect of Compound 25 or 64 on liver steatosis was evaluated by direct comparison of liver histology (Figure 15) between the drug-treated and vehicle-treated high- fat diet mice. The drug is highly efficacious in reversing fatty liver developed by HFD feeding. Figure 15 shows liver histology images with H&E stain from mice treated as noted: Figure 15A shows a normal liver section histology from a healthy C57B16 mouse without HFD feeding; Figure 15B shows liver section histology of a mouse fed HFD. Figure 15C and Figure 15D show liver sections from mice fed HFD following 3 weeks of treatment with Compounds 25 and 64, respectively (P.O. dosing of 5 mg/kg, by daily gavage).
|0631] Experiments were similarly performed with other compounds, in either high-fat diet induced diabetes and fatty liver models, or in db/db mouse model, or in both, by
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SUBSTITUTE SHEET ( RULE 26 ) daily oral gavage administration with low dosages. Table 9 shows the compounds, the animal model tested, and corresponding efficacious daily dose.
Table 9
|0632} Collectively, the data show that Compounds 25 and 64, and the other compounds tested, at low dosages (l-10mg/kg/day, by daily oral gavage administration), are highly efficacious in treating the above-mentioned symptoms in metabolic diseases in clinically relevant models. The efficacious dosages are greatly reduced as compared to prior disclosed compounds and the compounds do not require that the drug be mixed in food as compared to the prior disclosed compounds (International Patent Publication Number WO 2012/068274, International Patent Publication Number WO 2016/081599, U. S. Patent 10,227,315, and Tao et al., 2014).
Example B5
Inhibiting Hepatic Fibrosis In Vitro and In Vivo
[0633] Liver fibrosis is one hallmark of NASH. Liver fibrosis is mainly caused by hepatic stellate cells differentiating into myofibroblast cells upon stimulation by various inflammatory signals in response to liver damage, induced by steatosis and other factors. This process could be modeled in vitro using the hepatic stellate differentiation experiment. The human hepatic stellate LX-2 cells were cultured within DMEM (Dulbecco's Modified Eagle Media, pH 7.4) supplied with 2% fetal bovine serum, 4.5 g/1 glucose and 2 mM glutamine to 100 percent confluence. TGF0 (10 ng/ml) and Compound 25 (7.5 pM) or Compound 64 (1.0 pM) were then added into the media and incubated for 24 hours. Cellular morphology was then examined under a microscope. As shown in Figure 16 to Figure 17, hepatic stellate cells could be induced by TGF-beta to differentiate into myofibroblast-like cells. The experiments showed that treatment with Compound 25 and 64 effectively inhibit the differentiation process, indicating these compounds are efficacious in blocking stellate cell differentiation.
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SUBSTITUTE SHEET ( RULE 26 ) Figure 16B shows a panel of microscopic images of human hepatic stellate LX-2 cells in vehicle (DMSO), treated with TGFP which induces the differentiation of LX-2 cells into myofibroblast-like cells, which form the “ring”-shape morphology after TGFp treatment, and treated with TGFP and Compound 25 which prevents TGFP-induced differentiation of LX-2 cells. Figure 17A shows a panel of microscopic images of human hepatic stellate LX-2 cells treated with TGFP and Compound 64 which prevents TGFP-induced differentiation of LX-2 cells.
J06341 Some other example compounds tested and their effectiveness of inhibiting hepatic stellate cells differentiating to myofibroblast-like cells are summarized in Table 10.
Table 10
(06351 The molecular mechanism by which Compound 64 inhibits TGF-p signaling pathway was examined. Immunoblotting analysis showed Compound 64 blocks activation of TGF-P signaling pathway at early stages by inhibiting Smad2/3 phosphorylation (Figure 17B). LX-2 cells were treated with either vehicle (control), or TGF- P alone, or TGF- plus indicated concentrations of Compound 64 for 6 hrs. Cells were then harvested, and immunoblotting analyses was performed with antibodies against p-Smad2/3 (phosphorylated Smad2/3), Smad2/3, or GAPDH, as indicated.
[0636] Since Compounds 25, 64, 16, and 23, were able to directly inhibit hepatic stellate cell (HSC) activation in vitro, a direct in vivo anti-fibrotic effect test (independent of anti-steatosis effect) was conducted using the carbon tetrachloride (CCD-induced liverdamaging and liver fibrosis NASH murine models. CCL is a hepatotoxin that induces acute liver injury in mice and induces oxidative damage, inflammation, and liver fibrosis in mice, independent of liver steatosis, and thus is considered as a “pure” fibrotic model.
[0637] C57B16 male mice at the age of 7-8 weeks were randomized into 3 groups: vehicle, CCL alone treatment, CCL plus Compound treatment, n = 7. CCL was diluted in olive
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SUBSTITUTE SHEET ( RULE 26 ) oil (1 :5 v/v), and intraperitoneally injected at a dose of 750 pl/kg to the mice, twice a week for 6 weeks. Compound treatment started at the beginning of CCL injection. At the end of treatment, the mice were euthanized and liver tissue was collected. For histology analyses, the liver tissue was fixed and paraffin embedded; tissue sections were stained with hematoxylin and eosin (H&E), or with Picrosirius red. The fibrosis severities were scored by a board- certified pathologist based on Picrosirius red stain of the liver. For immunoblotting analyses, the liver tissue was homogenized and the proteins were separated by native page gel electrophoresis. The levels of Collagen I and GAPDH were detected by Collagen I polyclonal antibody (Abeam ab34710) and GAPDH (14C10) Rabbit mAb (Cell signaling #2118).
[0638| As shown in Figure 18, treatment with Compound 25 reduced fibrosis formation and improved fibrosis scores (Figure. 18 A-B). Moreover, molecular characterization by immunoblotting with collagen antibody confirmed that Compound 25 reduces collagen levels, the biomarker of fibrosis (Figure 18 C).
Example B6
Effect on Cancer Cell Growth Inhibition
[0639] Cell growth inhibition assays were performed using a protocol standardized by National Cancer Institute (NCI) for evaluating anti-cancer drugs. Various mammalian cell lines were used for the assays. Briefly, the cells were inoculated into 96 well microtiter plates. After cell inoculation, the microtiter plates were incubated for cell growth for 24 h prior to the addition of compounds. After 24 h, two plates of each cell line were fixed in situ with TCA, to represent a measurement of the cell population for each cell line at the time of compound addition (Tz). Test compounds were solubilized in dimethyl sulfoxide. At the time of test compound addition, five 10-fold or ’A log serial dilutions were made to provide a total of five test compound concentrations plus control. Aliquots of 100 pl of these different test compound dilutions were added to the appropriate microtiter wells already containing 100 pl of the medium, resulting in the required final test compound concentrations. Following test compound addition, the plates were incubated for an additional 48 h. The assay was terminated by the addition of cold TCA. Cells were fixed in situ with 50 pl of cold 50 % (w/v) TCA (final concentration, 10 % TCA), washed five times with tap water and air-dried. Sulforhodamine B (SRB) solution (100 pl) at 0.4 % (w/v) in 1 % acetic acid was added to each well, and plates were incubated for 10 minutes at room temperature. After staining, the unbound dye was removed by washing five times with 1 %
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SUBSTITUTE SHEET ( RULE 26 ) acetic acid and the plates were air-dried. The bound stain was subsequently solubilized with 10 mM trizma base, and the absorbance was read on an automated plate reader at a wavelength of 515 nm. Using the seven absorbance measurements [time zero, (Tz), control growth, (C), and test growth in the presence of a compound at the five concentration levels (Ti)], the percentage growth was calculated at each of the test compound concentrations levels. Growth inhibition of 50 % (GI50) was calculated from [(Ti-Tz)Z(C-Tz)] x 100 = 50, which is the test compound concentration resulting in a 50% reduction in the net protein increase (as measured by SRB staining) in control cells during the compound incubation. Table 11 shows the GI50 of the tested example compounds.
Table 11
1, GI50, 50% growth inhibition activity: ***, <5 pM; **>5 but < 20 pM; * >20 pM
Example B7
Metabolic Stability Assay
[0640| The metabolic stability of the compounds was evaluated in incubation with rat liver microsomes. Briefly, 1 pM of each testing compound was incubated within rat liver microsomes (from BD Gentest) with final liver microsomal protein concentration of 0.5 mg/ml for different time points (0, 5, 15, 30 and 45 min) at 37°C, in potassium phosphate buffer plus 6 mM NAPDH. After the incubation, the reactions were quenched and the remaining compound concentrations were then extracted and determined by LC- MS/MS.Results are described in Tables 4 and 5.
Example B8
Rat Pharmacokinetics Study
[064.1] Pharmacokinetics of the testing compounds was determined in male SD rats, n = 3. Briefly, each compound was prepared in 0.5% CMC -Na and 1% Tween in water to yield a homogeneous suspension at 1 mg/ml, and then administrated to 10 mg/kg via oral gavage. The animals were fasted overnight before dosing and fed for 4 hour post dosing with free access to water for all time. Blood samples were collected at pre-dose, 0.083, 0.25, 0.5,
-290-
SUBSTITUTE SHEET ( RULE 26 ) 1, 2, 4, 8 and 24 hours, into KzEDTA-coated tube via tail vein for serial sampling and cardiac puncture for terminal sampling. Blood samples were put on ice and centrifuged to obtain plasma samples (2000g, 5 min under 4°C) within 15 minutes. Plasma samples were stored at approximately -80°C until analysis. The compound in the plasma samples were then extracted and concentrations determined by LC-MS/MS.
Example B9
Determination of Acute Toxicity
|0642] Acute toxicity tests were performed on C57B16 mice, male, 6 weeks old, n=6 for each dose. Compounds 64 and 25 were prepared to fine suspension in 0.5% CMC-Na / 1% Tween80 water solution. The tests were performed under fed condition, by oral gavage of 100-400 pL compound suspension according to the body weight to desired dosages (mg/kg). The mice were supplied with drinking water all through the testing time. Mouse behaviors were monitored every 15-30 minutes. The dosage leading to the death of 50% of animals is defined as LD50. In Figure 8, LD50 of Compound 64 was measured at around 420 mg/kg, while the LD50 of Compound 25 was measured over 500mg/kg.
Example B10
Short-Term Toxicology Study
]0643] 10-day escalating toxicology study in CD-I mice: Different doses of compounds (e.g. Compound 25 and Compound 64) were given to male CD1 mice once daily via oral gavage, for 10 consecutive days. The compound was administered as a homogeneous suspension in 0.5% CMC + 1% Tween80 in water in a volume of 10 mL/kg for oral administration. There were two arms in the study: 5 mice in the toxicology group and 5 mice in the toxicokinetic (TK) satellite group Clinical signs were recorded twice daily. Body weight and food consumption were recorded once daily. Rectal temperature was recorded once every two days for a total of 5 times throughout the study. Necropsies were conducted at the termination of the study. Hematology, serum chemistry, plasma exposure, and tissue distribution were evaluated post 10th dosing in the TK satellite animals. Histopathology was examined in selected organs/tissues. Based on the outcomes of the toxicology parameters, NOAEL (no-observable-adverse-effect-level) and MTD (maximal tolerable dose) were determined.
-291-
SUBSTITUTE SHEET ( RULE 26 ) [06441 In Figure 9 and Figure 10, for Compound 25, 25mg/kg/day, 50 mg/kg/day, and 75 mg/kg/day were tested; for Compound 64, 20mg/kg/day, 30mg/kg/day, and 40 mg/kg/day were tested. The NOAEL of Compound 25 was 50 mg/kg/day, while the NAOEL of Compound 64 was 40 mg/kg/day.
Example B 11
TGF-P Inhibitory Effect in T cells
[0645] The effect of compounds (e.g. Compound 25 and Compound 64) on the TGF-P signaling pathway was analyzed in T- Cells. Jurkat cells (a human T-cell cancer cell line) and the primary mouse CD8+ cells (T-cells) were tested. The cells were treated with TGF-P (10 ng/ml) alone, or TGF-P (10 ng/ml) plus compound at various concentrations (e.g. 0.5, 1.0, 2.0 pM), or treated with vehicle (control), for 6 hours. The cells were then harvested, and cell lysates were subjected to immunoblotting analyses with antibodies against Smad2/3, phosphorylated Smad2/3, and GAPDH. As shown in Figure 19, Compound 64 effectively blocks TGF-P signaling pathway activation in T cells, as demonstrated by inhibiting Smad2/3 phosphorylation, an essential event during TGF-P activation.
Example B 12
Anti-Cancer Effect in Combination with a PD-1 Inhibitor
[0646] Cancer immunotherapy provides a powerful therapeutic strategy. For example, antibodies against PD-1 /PD-L1 and cell therapy using CAR-T have been proved efficacious in some cancers. However, many types of tumors do not respond to these immunoncology therapies. Moreover, within the same type of cancer, immunoncology therapies are not equally efficacious for all patients. One important variable that dictates the immunoncology treatment outcomes is the cancer microenvironment (CME). The various cancer metabolites in CME as well as the presence of high levels of TGF-P are known to inhibit T-cells which may otherwise be efficacious in eliminating cancer cells.
[0647[ Due to the strong inhibitory effect of Compound 64 on the TGF-P pathway, a combinatory treatment of anti-PDl antibody with Compound 64 was tested to determine the efficacy of the combination against metastatic liver cancer of various origins, such as colorectal origin.
[0648| The experimental design is illustrated in Figure 20A. Mouse colon cancer cells, MC38, were transplanted in livers of immune competent B57/B16 mice, and the effect
-292-
SUBSTITUTE SHEET ( RULE 26 ) of the combinatory treatment was determined. 100,000 or 250,000 MC38 cells suspended in 50 pl PBS were injected in the main lobe of the liver of C57B16 mice (female, 8 weeks old).
1 week after cancer cell transplantation, the mice were randomized in 3 groups for the following treatments: vehicle, anti-PD-1 antibody alone, and Compound 64 plus anti-PD-1 antibody. Compound 64 was prepared as fine suspension in 0.5% CMC -Na + 1% Tween80 water solution. Treatment was started from day 7, at a dose of 2.5 mg/kg, once per day by oral gavage. The InVivoPlus Anti-mouse PD-1 (CD279) antibody, and its control antibody, InVivoPlus rat IgG2a isotype control (BioXcell), were used. The antibodies were diluted using dilution buffer from the BioXcell. The Anti-PD-1 antibody and its control were administrated to the mice as 100 pg per mouse via i.p. injection for a total of 5 doses on day 8, 11, 14, 17 and 20 after the cancer cell implantation. The mice were euthanized on day 28. Tumor incidence and volume were determined.
|0649] As shown in Figure 20B, the combinatory treatment is highly efficacious as compared to vehicle or PD-1 antibody alone treated group, leading to the elimination of cancer in 90% of mice (90% cancer-free, P<0.01).
Example B 13
Anti-Viral Activity Against Enveloped Viruses, Using Coronavirus 229E and SARS-CoV-2 as Examples.
[0650] Replication and release of enveloped viruses require the host cells to synthesize lipids that support the assembly of viral envelopes, which consist of lipid bilayer. Mitochondrial uncouplers promote lipid oxidation and inhibit lipid synthesis. Thus, the compounds of the present invention were tested for antiviral activities against envelope viruses, using coronavirus 229E and SARS-CoV-2 (the pathogen of COVID 19) as examples.
[06511 Compounds 64, 25, and 57, were tested, and specificity index (SI) was determined. Inhibition of virus-induced cytopathic effects (CPE) and cell viability following SARS-CoV-2 (USA_WAl/2020 strain) replication or alpha coronavirus 229E replication in Vero 760 cells (SARS-COV-2) or MRC-5 cells (CoV-229E) was measured. Cells were seeded in 96-well flat-bottom tissue culture plates and allowed to adhere overnight.
Following overnight incubation, diluted test compounds and virus diluted to a pre-determined titer to yield 85 to 95% cell killing at 3 days (SARS CoV-2) or 6 days (CoV-229E) postinfection were added to the plate. Following incubation at 37°C, 5% CO2 for three or six days, cell viability was measured by Visual or sulforhodamine B staining. Percent CPE reduction of the virus-infected wells and the percent cell viability of uninfected drug control
-293-
SUBSTITUTE SHEET ( RULE 26 ) wells were calculated to determine the EC50 and TC50 values. Specificity Index (SI, ratio of TC50 over EC50) was calculated accordingly. As shown in Figure 21, Compounds 64, 25, and 57 exhibited anti-viral effects with SI ranging from 10-79.
|0652] Although the present disclosure has been described in considerable detail with reference to certain preferred versions thereof, other versions are possible. Therefore, the spirit and scope of the present disclosure should not be limited to the description of the preferred versions described herein.
[0653] Although compositions, materials, and methods similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable preparations, methods and materials are described herein. All publications mentioned herein are incorporated by reference in their entirety. In the case of conflict, the present specification, including definitions, will control. In addition, the particular embodiments discussed below are illustrative only and not intended to be limiting.
[0654] All features disclosed in the specification, including the abstract and drawings, and all the steps in any method or process disclosed, may be combined in any combination, except combinations where at least some of such features and/or steps are mutually exclusive. Each feature disclosed in the specification, including abstract and drawings, can be replaced by alternative features serving the same, equivalent or similar purpose, unless expressly stated otherwise. Thus, unless expressly stated otherwise, each feature disclosed is one example only of a generic series of equivalent or similar features. Various modifications of the present disclosure, in addition to those described herein, will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims.
-294-
SUBSTITUTE SHEET ( RULE 26 )

Claims

CLAIMS What Is Claimed Is :
1 . A compound of F ormul a A : wherein R1000a of Formula A is selected from the group consisting of
-CH3, -CH2CH3, -C1-C6 alkyl, -C3-C6 cycloalkyl, -0CH3, -CH2OCH3, -CH2OCH2OCH3,
C(O)N(CH2CH2OCH3)2; each of substituents R5000A andR5000B is independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH2)2OCH3; alternatively R5000A and R5000B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl;
R6000 is selected from the group consisting of a 5 to 6-membered heterocyclyl, pyridinyl and thiazolyl; r’ is an integer selected the group consisting of 1, 2 and 3; r is an integer selected from the group consisting of 0, 1, 2 and 3;
Rloooc of Formula I is selected from the group consisting of chloro, fluoro, iodo, and bromo;
-295-
SUBSTITUTE SHEET ( RULE 26 ) each of R4000b and R4000d is independently selected from the group consisting of Y1000 and Z1000, provided that when R4000b is Y1000, R4000d is z1000 and when R4000b is z1000, R4000d is yiooo.
Y1000 is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , - CHF2, fluoro(Ci-C6)alkyl, halo(Ci-C6)alkyl, -OCF3, -SO2(Ci-C6)alkyl, cyano, and -CO2(Ci- Ce)alkyl;
Z1000 is selected from the group consisting of H, -CH2OCH3, -CH2OCH2CH3,
R2OOOA ANC| R2OOOB together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclic ring optionally substituted with one or more methyl groups;
R3000 is a 5 to 6-membered heterocyclic ring;
Ar1 is a 5 to 6-membered aryl or heteroaryl group optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl, halo, hydroxyl and alkoxy; each of R7000A and R7000B is independently selected from Ci-Ce alkyl; alternatively R7OOOA anj R7000B together with the nitrogen to which they are attached, form a 4 to 8- membered heterocyclyl optionally substituted with one or more substituents independently selected from Ci-Ce alkyl;
R8000 is selected from the group consisting of a 5 to 6-membered heterocyclic ring optionally substituted with methyl; s is an integer selected from the group consisting of 0, 1, 2 and 3; t’ is an integer selected from the group consisting of 1, 2 and 3; t is an integer selected from the group consisting of 0, 1, 2 and 3; with the proviso that
R5000Aand R5000B are not both Ci-Ce alkyl; and
-296-
SUBSTITUTE SHEET ( RULE 26 ) when Z1000 is H; R1000a is not Ci- C6 alkyl, -C3-C6 cycloalkyl, CH3 or CH2CH3; or a pharmaceutically acceptable salt, solvate, or prodrug thereof,
2. A compound of F ormul a I : each of R5A andR5B is independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH2)2OCH3; alternatively R5A and R5B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl;R6 is selected from the group consisting of 5 to 6-membered heterocyclyl, pyridinyl and thiazolyl; m’ is an integer selected the group consisting of 1, 2 and 3; m is an integer selected the group consisting of 0, 1, 2 and 3;
-297-
SUBSTITUTE SHEET ( RULE 26 ) Rlc is selected from the group consisting of chloro, fluoro, iodo, and bromo; each of R4b and R4d is independently selected from the group consisting of Y and Z, provided that when R4b is Y, R4d is Z and when R4h is Z, R4d is Y;
Y is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-C6)alkyl, halo(Ci-C6)alkyl, -OCF3, -SO2(Ci-C6)alkyl, cyano, and -CO2(Ci-C6)alkyl;
Z is selected from the group consisting of -CH2OH, -CH2OCH2CH3, -CH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3, -CH2O(CH2)2NHCH3, -CH2O(CH2)2N(CH3)2, -CH2O(CH2)2NHSO2CH3, -(CH2)O(CH2)2NR2AR2B, -(CH2)nR3, -CH2OCH2Ar, and OCH3;
R2A and R2B together with the nitrogen to which they are attached, form a 4 to
8-membered heterocyclic ring optionally substituted with one or more methyl groups;
R3 is selected from the group consisting of a 5 to 6-membered heterocyclic ring and phenoxy; n is an integer selected from the group consisting of 0 1, 2, and 3; and
Ar is a 5-6 membered aryl or heteroaryl group optionally substituted with one or more substituents, independently selected from the group consisting of -Ci-Ce alkyl, halo, hydroxy, and alkoxy; or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
3. The compound of claim 2, wherein Rla is selected from the group consisting of -CH3, -CH2CH3 and -CH2NR5AR5B.
4. The compound of claim 3, wherein each of R5A and R5B is independently selected from Ci-Ce alkyl substituted with one or more methoxy; alternatively, R5A and R5B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl.
5. The compound of claim 2, wherein Rlais selected from the group consisting of
-298-
SUBSTITUTE SHEET ( RULE 26 )
6. The compound of claim 2 wherein Rlc is chloro.
7. The compound of claim 2 wherein R4b is Y and R4d is Z.
8. The compound of claim 2 wherein R2A and R2B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with a methyl group.
9. The compound of claim 2 wherein R2A and R2B together with the nitrogen to which they are attached, form a heterocyclyl selected from piperazinyl or a 4-methyl piperazinyl.
10. The compound of claim 2 wherein R3 is a 5-membered heterocyclic ring.
11. The compound of claim 2 wherein R3 is tetrahydrofuranyl.
12. The compound of claim 2 wherein n is 0.
13. The compound of claim 2 wherein -(CH2)nR is .
14. The compound of claim 2 wherein Ar is selected from the group consisting of a 5 to 6-membered aryl or heteroaryl optionally substituted with one or more substituents independently selected from the group consisting of methyl, fluoro, chloro, hydroxy and methoxy.
15. The compound of claim 14 wherein Ar is selected from the group consisting
16. The compound of claim 2 wherein Yis CF3
17. The compound of claim 2, wherein Z is selected from the group consisting of - CH2OCH2CH3,-CH2OCH3, -CH2O(CH2)2OH, -CH2O(CH2)2OCH3, -CH2O(CH2)2N(CH3)2J -CH2O(CH2)2NHSO2CH3, -(CH2)O(CH2)2NR2A R2B, -(CH2)mR3, -CH2OCH2Ar and OCH3; wherein R2A and R2B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of piperazinyl and 4-methyl piperazinyl
R3 is tetrahydrofuranyl; and
-299-
SUBSTITUTE SHEET ( RULE 26 ) Ar is a 5 to 6-membered aryl or heteroaryl group optionally substituted with one or more substituents independently selected from the group consisting of methyl, fluoro, chloro, hydroxyl and methoxy.
18. The compound of claim 2 wherein
R4d is selected from the group consisting of -CH2OCH2CH3, -CH2OCH3,
19. The compound of claim 2 wherein
Rlais selected from the group consisting of -CH3, -CH2CH3 and -CH2NR5AR5B; each of R5A and R5B is independently selected from Ci-Ce alkyl substituted with one or more methoxy; alternatively, R5A and R5B together with the nitrogen to which they are attached, form a 4-8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl;
Rlc is chloro
Y is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-Cg)alkyl, halo(Ci-Ce)alkyl, -OCF3, -SO2(Ci-Ce)alkyl, cyano, and -CC>2(Ci- C6)alkyl;
Z is selected from the group consisting of -CH2OCH2CH3, -CH2OCH3, - CH2O(CH2)2OH, -CH2O(CH2)2OCH3, -CH2O(CH2)2N(CH3)2, -CH2O(CH2)2NHSO2CH3, - (CH2)O(CH2)2 NR2A R2B, -(CH2)mR3, -CH2OCH2Ar and OCH3; wherein R2A and R2B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of piperazinyl and 4-methyl piperazinyl;
-300-
SUBSTITUTE SHEET ( RULE 26 ) R3 is tetrahydrofuranyl;
Ar is a 5 to 6-membered aryl or heteroaryl group optionally substituted with one or more substituents independently selected from the group consisting of methyl, fluoro, chloro, hydroxy and methoxy; or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
20. The compound of claim 2 wherein
Rlais selected from the group consisting of -CH3, -CH2CH3 and -CH2NR5AR5B; each of R5A and R5B is independently selected from Ci-Ce alkyl substituted with one or more groups selected from methoxy; alternatively, R5A and R5B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl;
Rlc is chloro
Y is CF3;
Z is selected from the group consisting of -CH2OCH2CH3,-CH2OCH , - CH2O(CH2)2OH, -CH2O(CH2)2OCH3, -CH2O(CH2)2N(CH3)2, -CH2O(CH2)2NHSO2CH3, - (CH2)O(CH2)2NR2A R2B, -(CH2)mR3, -CH2OCH2Ar and OCH3;
R2A and R2B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of piperazinyl or a 4-methyl piperazinyl
R3 is tetrahydrofuranyl;
Ar is a 5 to 6-membered aryl or heteroaryl group optionally substituted with one or more substituents independently selected from the group consisting of methyl, fluoro, chloro, hydroxyl, and methoxy or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
21. The compound of claim 2 wherein
-301-
SUBSTITUTE SHEET ( RULE 26 ) Rlais selected from the group consisting of -CH3, -CH2CH3, -
Rlc is chloro;
Y is CF3; and
Z is selected from the group consisting of -CH2OCH2CH3, -CH2OCH3, - or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
22. The compound of claim 21 wherein
R4b is Y and R4d is Z.
23. The compound of claim 2 of Formula la
-302-
SUBSTITUTE SHEET ( RULE 26 ) wherein Rlais selected from the group consisting of -CH3, -CH2CH3, -
R4d is selected from the group consisting of CH2OH, -CH2OCH2CH3, -CH2OCH3, or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
24. The compound of claim 23 wherein
R4d is selected from the group consisting of -CH2OCH2CH3, -CH2OCH3,
25. The compound of claim 2selected from the group consisting of:
-303-
SUBSTITUTE SHEET ( RULE 26 )
-304-
SUBSTITUTE SHEET (RULE 26)
-305-
SUBSTITUTE SHEET (RULE 26)
-306-
SUBSTITUTE SHEET (RULE 26)
-307-
SUBSTITUTE SHEET (RULE 26)
-308-
SUBSTITUTE SHEET (RULE 26)
-309-
SUBSTITUTE SHEET (RULE 26)
-310-
SUBSTITUTE SHEET (RULE 26) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
26. The compound of claim 25 that is
-311-
SUBSTITUTE SHEET ( RULE 26 ) pharmaceutically acceptable salt, solvate, or prodrug thereof.
27. A compound of Formula II: wherein:
R10a is selected from the group consisting of -OCH3, -CH2OCH3, -CH2OCH2OCH3,
C(O)N(CH2CH2OCH3)2; each of R50AandR50B is independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-Ce alkoxy, and -O(CH2)2OCH3; alternatively R50A and R50B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl;
-312-
SUBSTITUTE SHEET ( RULE 26 ) R60 is selected from the group consisting of 5 to 6-membered heterocyclyl, pyridinyl and thiazolyl; o’ is an integer selected from the group consisting of 1, 2, and 3; o is an integer selected from the group consisting of 0, 1, 2, and 3;
R10c is selected from the group consisting of chloro, fluoro, iodo, and bromo; and one of R40b and R40d is H and the other of R40b and R40d is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-Ce)alkyl, halo(Ci-Ce)alkyl, -OCF3, -SO2(Ci-C6)alkyl, cyano and -CO2(Ci-C6)alkyl; with the proviso that R50A and R50B are not both Ci-Ce alkyl; or a pharmaceutically acceptable salt, solvate, or prodrug thereof
28. The compound of claim 27 wherein R10a is selected from the group consisting C(O)N(CH2CH2OCH3)2
29. The compound of claim 27, wherein each of R50A andR50B is independently selected from the group consisting of methyl; and Ci-Ce alkyl substituted with one or more groups selected from methoxy, and -CXCFL^OCFF; alternatively R50A and R50B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl selected from the group consisting of azetindinyl; pyrrolidinyl; pyrrolidinyl substituted with one or more substituents selected from the group consisting of cyano, hydroxyl, methoxy, -NHC(O)CH3,
-C(O)NH2, -SO2CH3, -CF3 and methyl; piperidinyl; piperidinyl substituted with one or more substituents selected from the group consisting of -CF3, and fluoro; piperazinyl substituted with one or more substituents selected from the group consisting of oxo, methyl, and acetyl; morpholinyl; morpholinyl substituted with one or more methyl groups; dioxothiomorphylinyl.
30. The compound of claim 29 wherein -CH2NR50AR50B is selected from the group consisting of the group consisting of -CH2N(CH3)(CH2)2OCH3, -(CH2)N(CH2CH2OCH3)2, -
-313-
SUBSTITUTE SHEET ( RULE 26 )
31. The compound of claim 27 wherein R60 is selected from the group consisting of tetrahydrofuranyl, thiazolyl, and pyridinyl and o is 0.
32. The compound of claim 27 wherein -(CH2)o R‘ is selected from the group consisting
33. The compound of claim 27 wherein R10a is selected from the group consisting
-314-
SUBSTITUTE SHEET ( RULE 26 )
and C(O)N(CH2CH2OCH3)2.
34. The compound of claim 27 wherein one of R40b and R40d is H and the other of R40b and R40d is selected from the group consisting of fluoro, -CF3 , -CHF2, and -OCF3.
35. The compound of claim 34 wherein R40d is H and R40b is selected from the group consisting of fluoro, -CF3 , -CHF2, and -OCF3.
36. The compound of claim 27, wherein
-315-
SUBSTITUTE SHEET ( RULE 26 )
C(O)N(CH2CH2OCH3)2;
R10c is selected from the group consisting of chloro, fluoro, iodo; and
R40d is H and R40b is selected from the group consisting of fluoro, -CF3, -CHF2, and -OCF3; or a pharmaceutically acceptable salt, solvate, or prodrug thereof
37. The compound of claim 36 wherein R10A is selected from the group consisting
-316-
SUBSTITUTE SHEET ( RULE 26 )
38. The compound of claim 37 wherein R10c is chloro; and R40b is -CF3.
39. The compound of claim 27 of Formula Ila
-317-
SUBSTITUTE SHEET ( RULE 26 )
C(O)N(CH2CH2OCH3)2; or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
40. The compound of claim 39 wherein R10a is selected from the group consisting
-318-
SUBSTITUTE SHEET ( RULE 26 )
-C(O)N(CH2CH2OCH3)2.
41. A compound selected from the group consisting of:
-319-
SUBSTITUTE SHEET ( RULE 26 )
-320-
SUBSTITUTE SHEET (RULE 26)
-321-
SUBSTITUTE SHEET (RULE 26)
-322-
SUBSTITUTE SHEET (RULE 26)
-323-
SUBSTITUTE SHEET (RULE 26)
-324-
SUBSTITUTE SHEET (RULE 26)
-325-
SUBSTITUTE SHEET (RULE 26)
-326-
SUBSTITUTE SHEET (RULE 26) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
42. The compound of claim 41 selected from the group consisting of pharmaceutically acceptable salt, solvate, or prodrug thereof.
43. A compound of Formula III: wherein:
-327-
SUBSTITUTE SHEET ( RULE 26 ) R100a is selected from the group consisting of -CH3, -OCH3, -CH2OCH3, each of R500A andR500B is independently selected from the group consisting of -Ci-Ce alkyl; and -Ci-Ce alkyl substituted with one or more groups selected from -Ci-G> alkoxy, and -O(CH2)2OCH3; alternatively R500A and R500B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more substituents, independently selected from the group consisting of oxo, cyano, hydroxyl, alkoxy, acylamino , carboxyamido, -SO2CH3, -CF3, Ci-Ce alkyl, halo, and acyl;
R600 is selected from the group consisting of a 5 to 6-membered heterocyclyl, pyridinyl and thiazolyl; p’ is an integer selected from the group consisting of 1, 2, and 3; p is an integer selected the group consisting of 0, 1, 2 and 3;
R100c is selected from the group consisting of chloro, fluoro, iodo, and bromo; each of R400b and R400d is independently selected from the group consisting of Y1 and Z1, provided that when R400b is Y1, R400d is Z1 and when R400b is Z1, R400d is Y1;
Y1 is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-C6)alkyl, halo(Ci-Ce)alkyl, -OCF3, -SO2(Ci-Ce)alkyl, cyano, and -CO2(Ci-C6)alkyl;
Z1 is selected from the group consisting of -CH2NHC(O)CH2CH3, -CH2NHC(O)CH2OCH3, -CH2NHSO2CH3, -(CH2)q NR7AR7B, and -(CH2)qR8; each ofR7A andR7B is independently selected from Ci-Ce alkyl; alternatively R7A and R7B together with the nitrogen to which they are attached, form a 4 to 8-membered heterocyclyl optionally substituted with one or more independently selected Ci-Ce alkyl;
R8 is selected from the group consisting of a 5 to 6-membered heterocyclic ring optionally substituted with methyl;
-328-
SUBSTITUTE SHEET ( RULE 26 ) q' is an integer selected from the group consisting of 1, 2 and 3; and q is an integer selected from the group consisting of 0, 1, 2 and 3; or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
44. The compound of claim 43 wherein R100a is selected from the group consisting of -CH3, and -CH2NR500AR500B
45. The compound of claim 44 wherein each of R500A and R500B is independently selected from Ci-Ce alkyl substituted with one or more methoxy; alternatively, R500A and R500B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl.
46. The compound of claim 43 wherein R100a is selected from the group consisting
47. The compound of claim 43 wherein R100c is chloro.
48. The compound of claim 43 wherein R400b is Y and R400dis Z.
49. The compound of claim 43 wherein each of R7A and R7B is methyl; alternatively, R7A and R7B together with the nitrogen to which they are attached, form a
6-membered heterocyclyl optionally substituted with a methyl group.
50. The compound of claim 43 wherein R7A and R7B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of, 4-methylpiperazinyl, and morpholinyl.
51. The compound of claim 43 wherein R8 is a 6-membered heterocyclic ring optionally substituted with methyl.
52. The compound of claim 43 wherein R8 is 4-methylpiperidinyl.
53. The compound of claim 43 wherein q is 0.
54. The compound of claim 43 wherein Z1 is selected from the group consisting of -CH2NHC(O)CH2CH3, -CH2NHC(O)CH2OCH3,-CH2NHSO2CH3, -CH2NR7AR7B, and -(CH2)qR8;
-329-
SUBSTITUTE SHEET ( RULE 26 ) wherein each of R7A and R7B is methyl; alternatively R7A and R7B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of,
4-methylpiperazinyl, and morpholinyl; and and R8 is 4-methylpiperidinyl.
55. The compound of claim 43 wherein Z1 is selected from the group consisting of
56. The compound of claim 43 wherein Y1 is CF3.
57. The compound of claim 43 wherein is Z1 is selected from the group consisting
58. The compound of claim 43 wherein:
R100ais selected from the group consisting of -CH3, and -CH2NR500AR500B; each of R500Aand R500B is independently selected from Ci-Ce alkyl substituted with one or more methoxy; alternatively, R500A and R500B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl;
R100c is chloro;
Y1 is selected from the group consisting of chloro, fluoro, iodo, bromo, -CF3 , -CHF2, fluoro(Ci-C6)alkyl, halo(Ci-Ce)alkyl, -OCF3, -SO2(Ci-C6)alkyl, cyano and -CO2(Ci-C6)alkyl;
Z1 is selected from the group consisting of -CH2NHC(O)CH2OCH3, -CH2NHC(O)CH2CH3, -CH2NHSO2CH3, -CH2NR7AR7B, and -(CH2)qR8;
-330-
SUBSTITUTE SHEET ( RULE 26 ) each of R7A and R7B is methyl; alternatively R7A and R7B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of, 4-methylpiperazinyl, and morpholinyl; and
R8 is 4-methylpiperidinyl; or a pharmaceutically acceptable salt, solvate, or prodrug thereof
59. The compound of claim 43 wherein:
R100ais selected from the group consisting of -CH3, and -CH2NR500AR500B; each of R500Aand R500B is independently selected from Ci-Ce alkyl substituted with one or more methoxy; alternatively, R500A and R500B together with the nitrogen to which they are attached, form a 6-membered heterocyclyl optionally substituted with one or more substituents, independently selected from Ci-Ce alkyl;
R1°°C is chloro-
Y1 is -CF3;
Z1 is selected from the group consisting of -CH2NHC(O)CH2OCH3, -CH2NHC(O)CH2CH3, -CH2NHSO2CH3, -CH2NR7AR7B, and -(CH2)qR8; each of R7A and R7B is methyl; alternatively R7A and R7B together with the nitrogen to which they are attached, form a heterocyclyl selected from the group consisting of, 4-methylpiperazinyl, and morpholinyl; and
R8 is 4-methylpiperidinyl; or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
60. The compound of claim 43 wherein
R100ais selected from the group consisting of -CH3, -(CH2)N(CH2CH2OCH3)2,
RI°°C js chioj-o-
-331-
SUBSTITUTE SHEET ( RULE 26 ) Y1 is CF3; and or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
61. The compound of claim 60 wherein R400b is Y1 and R400d is Z1.
62. The compound of claim 43 of Formula Illa
R100ais selected from the group consisting of -CH3, -(CH2)N(CH2CH2OCH3)2, or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
63. The compound of claim 43 selected from the group consisting of
-332-
SUBSTITUTE SHEET ( RULE 26 )
-333-
SUBSTITUTE SHEET (RULE 26)
-334-
SUBSTITUTE SHEET (RULE 26) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.
64. A pharmaceutical composition comprising a compound of any one of claims 1 to 63 or a pharmaceutically acceptable salt or prodrug thereof and a pharmaceutically acceptable carrier or diluent.
65. A method of treating a mitochondria-related condition or disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition according claim 64.
66. The method of claim 65 wherein the mitochondria-related condition or disorder has and one or more underlying causal factors selected from the group consisting of hyperglycemia, abnormal accumulation of lipid in cells, abnormal accumulation of lipid in tissues, abnormal lipid metabolism, abnormal mitochondrial metabolism, insulin resistance, abnormal cell proliferation, abnormal TGF-beta activation, and abnormal fibrosis.
67. The method of claim 65 wherein the mitochondria-related condition or disorder has and one or more underlying symptoms selected from the group consisting of hyperglycemia, abnormal accumulation of lipid in cells, a bnormal accumulation of lipid in tissues, abnormal lipid metabolism, abnormal mitochondrial metabolism, insulin resistance, abnormal cell proliferation, abnormal TGF-beta activation, and abnormal fibrosis.
68. The method of any one of claims 65 to 67, wherein the mitochondria-related condition or disorder is selected from the group consisting of a metabolic disease, cancer, an autoimmune disease, pulmonary fibrosis, a dermatological disorder, an infectious disease, and a neurodegenerative disease.
69. The method of claim of 68, wherein the metabolic disease is selected from the group consisting of type 2 diabetes, a disease characterized by insulin resistance or hyperglycemia; obesity or obesity related complications, and a disease characterized by abnormal lipid accumulation.
SUBSTITUTE SHEET ( RULE 26 )
70. The method of claim 68, wherein the metabolic disease is a complication caused by type 2 diabetes, selected from the group consisting of diabetes-induced cardiovascular diseases, neurodegenerative disorders, atherosclerosis, hypertension, coronary heart diseases, nephropathy, retinopathy, neuropathy and diabetic heart failure.
71. The method of claim 68, wherein the metabolic disease or disorder is nonalcoholic fatty liver disease (NAFLD), wherein at least one prognosis stage of the disease is selected from the group consisting of hepatic steatosis, non-alcoholic steatohepatitis (NASH), cirrhosis, and NAFLD induced hepatocellular carcinoma (HCC).
72. The method of claim 68, wherein the metabolic disease or disorder is alcoholic fatty liver disease, or one or more complications caused by alcoholic fatty liver disease, wherein the one or more complications caused by alcoholic fatty liver disease is selected from the group consisting of alcoholic hepatitis, cirrhosis, and a combination thereof.
73. The method of claim 68, wherein the metabolic disease or disorder is dyslipidemia, or one or more complications caused by dyslipidemia.
74. The method of claim 68, wherein the cancer is a primary cancer selected from the group consisting of hepatocellular carcinoma, colorectal carcinoma, pancreatic cancer, breast cancer, prostate cancer, leukemia, lymphoma, melanoma, ovarian cancer, and lung cancer.
75. The method of claim 68, wherein the cancer is a metastatic cancer originated from a primary tumor of other tissue types.
76. The method of claim 68, wherein the pharmaceutical composition is administered in combination with a second agent indicated for the metabolic disease.
77. The method of claim 76, wherein the second agent is an anti-diabetic agent selected from the group consisting of metformin, insulin, insulin analogs, sulfonylureas, biguanides, meglit-inides, thiazolidinediones, alpha glucosidase inhibitors, GLP-1 agonists, DPP -4 inhibitors and SGLT2 inhibitors.
78. The method of claim 76, wherein the second agent is selected from the group consisting of an anti -obesity agent, an anti-nonalcoholic fatty liver disease agent, an anti- nonalcoholic fatty liver disease agent and an anti -dyslipidemia agent.
79. The method of either of claims 74 or 75, wherein the pharmaceutical composition is administered in combination with a second anti-cancer agent or anti-cancer regimen.
-336-
SUBSTITUTE SHEET ( RULE 26 )
80. The method of claim 79, wherein the second anti-cancer agent is an immunoncological agent.
81. The method of claim 80 wherein the immunocological agent is selected from the group consisting of an antibody against PD-1/PD-L1, an antibody against other immune check point proteins, CAR-T cells, and other therapeutic immune cells.
82. The method of claim 68, wherein the dermatological disorder is selected from eczema, dyshidrotic eczema, seborrheic eczema psoriasis, rosacea, dermatitis and atopic dermatitis.
83. The method of claim 68, wherein the infectious disease is a bacterial infection
84. The method of claim 68, wherein the infectious disease is viral infection.
85. The method of claim 84, wherein the viral infection is selected from SARS-
CoV-2 infection, a corona viral infection, and Ebola viral infection.
86. A method of treating a metabolic disease or disorder characterized by hyperglycemia or insulin resistance, or by abnormal accumulation of lipid in tissue, or a disease or a disorder in which hyperglycemia or insulin resistance or abnormal accumulation of lipid in tissue is a symptom, in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition according claim 64.
87. A method of treating cancer or hyperplasia, in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition according to claim 64.
88. A method of treating or preventing autoimmune diseases in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition of claim 64.
89. A method of treating or preventing a dermatological disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition according to claim 64.
90. A method of treating fibrosis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition according to claim 64.
-337-
SUBSTITUTE SHEET ( RULE 26 )
91. A method of treating or preventing a bacterial infection in a subj ect in need thereof, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition according to claim 64.
92. A method of treating or preventing a viral infection in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition according to claim 64
93. The method of claim of any of claims 65 to 92, wherein the subject is a mammal or a human.
94. The method of claim of claim 93, wherein the subject is a human.
95. The method of claim of any one of claims 65 to 94, wherein the pharmaceutical composition is administered orally, intravenously, subcutaneously, intramuscularly, transdermally, intraperitoneally, or by other pharmacologically acceptable routes.
96. A method for long-term disease management of a metabolic disease or disorder, or for long-term disease management of cancer, comprising administering to a subject in need thereof in need of such long-term management an effective amount of a pharmaceutical composition according to claim 64.
97. Use of a compound of any one of claims 1 to 63 or a pharmaceutically acceptable salt or prodrug thereof in the manufacture of a medicament for treatment of a metabolic disease or disorder characterized by hyperglycemia, or insulin resistance or by abnormal accumulation of lipid in tissue; a disease or a disorder in which hyperglycemia, or insulin resistance or abnormal accumulation of lipid in tissue is a symptom; cancer; hyperplasia; or cancer or hyperplasia related complications; diabetes; obesity; non-alcoholic fatty liver disease; alcoholic fatty liver disease; dyslipidemia; a dermatological disorder; or a bacterial infection; or a viral infection.
98. A method of preparing a mitochondrial membrane-retaining mitochondrial uncoupler comprising:
1. identifying a conventional mitochondrial uncoupler;
2. designing a compound that covalently links at least one secondary or tertiary amino moiety to a conventional mitochondrial uncoupler; and
-338-
SUBSTITUTE SHEET ( RULE 26 )
3. preparing the compound of step 2, wherein the compound is a mitochondrial membrane retaining uncoupler compound.
99. A mitochondrial membrane-retaining uncoupler compound of the Formula:
(RA)U-RB; or a pharmaceutically acceptable salt, solvate or prodrug thereof; wherein RAand RB are covalently linked; each RA is independently a moiety containing a secondary or tertiary amine; u is an integer selected from the group consisting of 1 and 2; and
RB is a conventional mitochondrial uncoupler prior to being covalently linked to RA; provided the mitochondrial membrane-retaining uncoupler compound is not a pharmaceutically acceptable salt, solvate, or prodrug thereof.
100. The compound of claim 97 wherein RA is selected from the group consisting of: RA is selected from the group consisting of -CH2NHSO2CH3, -CH2N(CHs)2, -(CH2)2N(CH3)2, -
-339-
SUBSTITUTE SHEET ( RULE 26 )
-340-
SUBSTITUTE SHEET (RULE 26)
EP23863911.6A 2022-09-08 2023-08-28 Mitochondrial uncouplers for treatment of metabolic diseases and cancer Pending EP4583870A2 (en)

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