WO2021247921A1 - Benzothiazole compounds and uses thereof - Google Patents

Benzothiazole compounds and uses thereof Download PDF

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WO2021247921A1
WO2021247921A1 PCT/US2021/035778 US2021035778W WO2021247921A1 WO 2021247921 A1 WO2021247921 A1 WO 2021247921A1 US 2021035778 W US2021035778 W US 2021035778W WO 2021247921 A1 WO2021247921 A1 WO 2021247921A1
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weeks
mmol
pyridin
thiazol
compound
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PCT/US2021/035778
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French (fr)
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WO2021247921A8 (en
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Bhaumik PANDYA
Madeline MACDONNELL
Kerem OZBOYA
Iwona WRONA
Vanessa KURIA
Bertrand Le Bourdonnec
Matthew Lucas
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Yumanity Therapeutics, Inc.
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Publication of WO2021247921A1 publication Critical patent/WO2021247921A1/en
Publication of WO2021247921A8 publication Critical patent/WO2021247921A8/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D213/36—Radicals substituted by singly-bound nitrogen atoms
    • C07D213/40—Acylated substituent nitrogen atom
    • 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
    • C07D417/12—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
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
    • C07D513/04—Ortho-condensed systems

Definitions

  • TDP-43 is a nuclear DNA/RNA binding protein involved in RNA splicing. Under pathological cell stress, TDP-43 translocates to the cytoplasm and aggregates into stress granules. These phenotypes are hallmarks of degenerating motor neurons and are found in 97% of all ALS cases. The highly penetrant nature of this pathology indicates that TDP-43 is broadly involved in both familial and sporadic ALS. Additionally, TDP-43 mutations that promote aggregation are linked to higher risk of developing ALS, suggesting protein misfolding and aggregation act as drivers of toxicity. TDP-43 toxicity can be recapitulated in yeast models, where the protein induces a viability deficit and localizes to stress granules.
  • the present inventors have discovered that the CYP51A1 inhibitors described herein are capable of reversing TDP-43 induced toxicity. Accordingly, the present invention describes such CYP51A1 inhibitors and methods of using these compounds for the treatment of disorders related to TDP-43 toxicity such as ALS.
  • the invention features a compound, or a pharmaceutically acceptable salt thereof, having the structure: Formula I wherein n is 0, 1, 2, 3, or 4; X is S or O; each R 1 is independently halo, cyano, hydroxy, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, -SO2-optionally substituted C1-C6 alkyl, or -CO2-optionally substituted C1-C6 alkyl; R 2 is optionally substituted C2-C9 heteroaryl; and R 3 is optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C9 heterocyclyl, optionally substituted C1-C6 alkyl C3-C8 cycloalkyl, optionally substituted C1-C6 heteroalkyl C3-C8 cycloalkyl, optionally substituted C1-C6 alkyl C2-C9 heteroaryl, optionally substituted C1
  • X is S. In some embodiments, X is O. In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, the compound has the structure: . Formula Ia In some embodiments, the compound has the structure: . Formula Ib In some embodiments, the compound has the structure: . Formula Ic In some embodiments, the compound has the structure: . Formula Id In some embodiments, at least one R 1 is halo (e.g., fluoro, chloro, or bromo). In some embodiments, at least one R 1 is cyano. In some embodiments, at least one R 1 is hydroxy.
  • At least one R 1 is optionally substituted C1-C6 alkyl (e.g., methyl, ethyl, trifluoromethyl, or hydroxymethyl). In some embodiments, at least one R 1 is optionally substituted C1-C6 heteroalkyl (e.g., ethoxy or trifluoromethoxy). In some embodiments, at least one R 1 is -SO2-optionally substituted C1-C6 alkyl (e.g., -SO2-methyl). In some embodiments, at least one R 1 is -CO2-optionally substituted C1-C6 alkyl (e.g., -CO2-methyl or -CO2-ethyl).
  • R 2 is a 5-membered optionally substituted C2-C9 heteroaryl (e.g., some embodiments, R 2 is a 6-membered optionally substituted C2-C9 heteroaryl (e.g., , , , In some embodiments, R 3 is optionally substituted C3-C8 cycloalkyl (e.g., cyclohexyl).
  • R 3 is optionally substituted C2-C9 heterocyclyl (e.g., , , , , , or , wherein R 4 is -C(O)R 5 ; R 5 is optionally substituted C1- C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C1-C6 alkyl C3-C8 cycloalkyl, or optionally substituted C1-C6 heteroalkyl C3-C8 cycloalkyl).
  • R 5 is optionally substituted C1- C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C1-C6 alkyl C3-C8 cycloalkyl, or optionally substituted C1-C6 hetero
  • R 3 is optionally substituted C6-C10 aryl (e.g., phenyl, 4-cyano-phenyl, or 4-fluoro-phenyl).
  • the invention features a compound, or a pharmaceutically acceptable salt thereof, having the structure of any one of compounds 1-273 in Table 1.
  • the invention features a pharmaceutical composition comprising any of the foregoing compounds and a pharmaceutically acceptable excipient.
  • the invention features a method of treating a neurological disorder (e.g., frontotemporal dementia (FTLD-TDP), chronic traumatic encephalopathy, ALS, Alzheimer’s disease, limbic-predominant age-related TDP-42 encephalopathy (LATE), or frontotemporal lobar degeneration) in a subject in need thereof.
  • a neurological disorder e.g., frontotemporal dementia (FTLD-TDP), chronic traumatic encephalopathy, ALS, Alzheimer’s disease, limbic-predominant age-related TDP-42 encephalopathy (LATE), or frontotemporal lobar degeneration
  • This method includes administering an effective amount of any of the foregoing compounds or pharmaceutical compositions.
  • the invention features a method of inhibiting toxicity in a cell (e.g., mammalian neural cell) related to a protein (e.g., TDP-43). This method includes administering an effective amount of any of the foregoing compounds or pharmaceutical compositions.
  • the invention features a method of treating a CYP51A1-associated disorder (e.g., FTLD-TDP, chronic traumatic encephalopathy, ALS, Alzheimer’s disease, LATE, or frontotemporal lobar degeneration) in a subject in need thereof. This method includes administering an effective amount of any of the foregoing compounds pharmaceutical compositions.
  • the invention features a method of inhibiting CYP51A1.
  • the invention features a method of treating a neurological disorder in a patient, such as a human patient, identified as likely to benefit from treatment with a CYP51A1 inhibitor on the basis of TDP-43 aggregation.
  • the method may include (i) determining that the patient exhibits, or is prone to develop, TDP-43 aggregation, and (ii) providing to the patient a therapeutically effective amount of a CYP51A1 inhibitor.
  • the patient has previously been determined to exhibit, or to be prone to developing, TDP-43 aggregation, and the method includes providing to the patient a therapeutically effective amount of a CYP51A1 inhibitor.
  • the susceptibility of the patient to developing TDP-43 aggregation may be determined, e.g., by determining whether the patient expresses a mutant isoform of TDP-43 containing a mutation that is associated with TDP-43 aggregation and toxicity, such as a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D.
  • the method includes the step of obtaining the sample from the patient.
  • the invention features a method of treating a neurological disorder in a patient, such as a human patient, identified as likely to benefit from treatment with a CYP51A1 inhibitor on the basis of TDP-43 expression.
  • the method includes (i) determining that the patient expresses a mutant form of TDP-43 having a mutation associated with TDP-43 aggregation (e.g., a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D), and (ii) providing to the patient a therapeutically effective amount of a CYP51A1 inhibitor.
  • a mutant form of TDP-43 having a mutation associated with TDP-43 aggregation e.g., a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D
  • the patient has previously been determined to express a mutant form of TDP-43 having a mutation associated with TDP- 43 aggregation, such as a Q331K, M337V, Q343R, N345K, R361S, or N390D mutation, and the method includes providing to the patient a therapeutically effective amount of a CYP51A1 inhibitor.
  • a mutation associated with TDP- 43 aggregation such as a Q331K, M337V, Q343R, N345K, R361S, or N390D mutation
  • the invention features a method of determining whether a patient (e.g., a human patient) having a neurological disorder is likely to benefit from treatment with a CYP51A1 inhibitor by (i) determining whether the patient exhibits, or is prone to develop, TDP-43 aggregation and (ii) identifying the patient as likely to benefit from treatment with a CYP51A1 inhibitor if the patient exhibits, or is prone to develop, TDP-43 aggregation.
  • the method further includes the step of (iii) informing the patient whether he or she is likely to benefit from treatment with a CYP51A1 inhibitor.
  • the susceptibility of the patient to developing TDP-43 aggregation may be determined, e.g., by determining whether the patient expresses a mutant isoform of TDP-43 containing a mutation that is associated with TDP-43 aggregation and toxicity, such as a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D. This may be performed, for example, by determining the amino acid sequence of a TDP-43 isoform isolated from a sample obtained from the patient or by determining the nucleic acid sequence of a TDP-43 gene isolated from a sample obtained from the patient. In some embodiments, the method includes the step of obtaining the sample from the patient.
  • the invention features a method of determining whether a patient (e.g., a human patient) having a neurological disorder is likely to benefit from treatment with a CYP51A1 inhibitor by (i) determining whether the patient expresses a TDP-43 mutant having a mutation associated with TDP-43 aggregation (e.g., a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D) and (ii) identifying the patient as likely to benefit from treatment with a CYP51A1 inhibitor if the patient expresses a TDP-43 mutant.
  • a mutation associated with TDP-43 aggregation e.g., a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D
  • the method further includes the step of (iii) informing the patient whether he or she is likely to benefit from treatment with a CYP51A1 inhibitor.
  • the TDP-43 isoform expressed by the patient may be assessed, for example, by isolated TDP-43 protein from a sample obtained from the patient and sequencing the protein using molecular biology techniques described herein or known in the art.
  • the TDP-43 isoform expressed by the patient is determined by analyzing the patient’s genotype at the TDP-43 locus, for example, by sequencing the TDP-43 gene in a sample obtained from the patient.
  • the method includes the step of obtaining the sample from the patient.
  • the CYP51A1 inhibitor is provided to the patient by administration of the CYP51A1 inhibitor to the patient. In some embodiments, the CYP51A1 inhibitor is provided to the patient by administration of a prodrug that is converted in vivo to the CYP51A1 inhibitor.
  • the neurological disorder is a neuromuscular disorder, such as a neuromuscular disorder selected from amyotrophic lateral sclerosis, congenital myasthenic syndrome, congenital myopathy, cramp fasciculation syndrome, Duchenne muscular dystrophy, glycogen storage disease type II, hereditary spastic paraplegia, inclusion body myositis, Isaac's Syndrome, Kearns-Sayre syndrome, Lambert–Eaton myasthenic syndrome, mitochondrial myopathy, muscular dystrophy, myasthenia gravis, myotonic dystrophy, peripheral neuropathy, spinal and bulbar muscular atrophy, spinal muscular atrophy, Stiff person syndrome, Troyer syndrome, and Guillain– Barré syndrome.
  • a neuromuscular disorder selected from amyotrophic lateral sclerosis, congenital myasthenic syndrome, congenital myopathy, cramp fasciculation syndrome, Duchenne muscular dystrophy, glycogen storage disease type II, hereditary spastic paraplegia, inclusion body myositis
  • the neurological disorder is amyotrophic lateral sclerosis.
  • the neurological disorder is selected from frontotemporal degeneration (also referred to as frontotemporal lobar degeneration and frontotemporal dementia), Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’s disease, Inclusion body myopathy with early-onset Paget disease and frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease, and hereditary inclusion body myopathy.
  • frontotemporal degeneration also referred to as frontotemporal lobar degeneration and frontotemporal dementia
  • Alzheimer’s disease Parkinson’s disease
  • dementia with Lewy Bodies corticobasal degeneration
  • progressive supranuclear palsy dementia parkinsonism ALS complex of Guam
  • the neurological disorder is amyotrophic lateral sclerosis
  • the patient exhibits one or more, or all, of the following responses: (i) an improvement in condition as assessed using the amyotrophic lateral sclerosis functional rating scale (ALSFRS) or the revised ALSFRS (ALSFRS-R), such as an improvement in the patient’s ALSFRS or ALSFRS-R score within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an improvement in the patient’s ALSFRS or ALSFRS-R score within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days
  • tautomeric forms result from the swapping of a single bond with an adjacent double bond and the concomitant migration of a proton.
  • a tautomeric form may be a prototropic tautomer, which is an isomeric protonation states having the same empirical formula and total charge as a reference form.
  • moieties with prototropic tautomeric forms are ketone – enol pairs, amide – imidic acid pairs, lactam – lactim pairs, amide – imidic acid pairs, enamine – imine pairs, and annular forms where a proton can occupy two or more positions of a heterocyclic system, such as, 1H- and 3H-imidazole, 1H-, 2H- and 4H- 1,2,4-triazole, 1H- and 2H- isoindole, and 1H- and 2H-pyrazole.
  • tautomeric forms can be in equilibrium or sterically locked into one form by appropriate substitution.
  • tautomeric forms result from acetal interconversion, e.g., the interconversion illustrated in the scheme below: .
  • isotopes of compounds described herein may be prepared and/or utilized in accordance with the present invention. “Isotopes” refers to atoms having the same atomic number but different mass numbers resulting from a different number of neutrons in the nuclei.
  • isotopes of hydrogen include tritium and deuterium.
  • an isotopic substitution may alter the physiciochemical properties of the molecules, such as metabolism and/or the rate of racemization of a chiral center.
  • many chemical entities in particular many organic molecules and/or many small molecules
  • can adopt a variety of different solid forms such as, for example, amorphous forms and/or crystalline forms (e.g., polymorphs, hydrates, solvates, etc).
  • such entities may be utilized in any form, including in any solid form.
  • such entities are utilized in a particular form, for example in a particular solid form.
  • compounds described and/or depicted herein may be provided and/or utilized in salt form. In certain embodiments, compounds described and/or depicted herein may be provided and/or utilized in hydrate or solvate form.
  • substituents of compounds of the present disclosure are disclosed in groups or in ranges. It is specifically intended that the present disclosure include each and every individual subcombination of the members of such groups and ranges.
  • the term “C 1 -C 6 alkyl” is specifically intended to individually disclose methyl, ethyl, C 3 alkyl, C 4 alkyl, C5 alkyl, and C6 alkyl.
  • a compound includes a plurality of positions at which substitutes are disclosed in groups or in ranges, unless otherwise indicated, the present disclosure is intended to cover individual compounds and groups of compounds (e.g., genera and subgenera) containing each and every individual subcombination of members at each position.
  • a phrase of the form “optionally substituted X” e.g., optionally substituted alkyl
  • X is optionally substituted
  • alkyl wherein said alkyl is optionally substituted
  • acyl represents a hydrogen or an alkyl group, as defined herein that is attached to a parent molecular group through a carbonyl group, as defined herein, and is exemplified by formyl (i.e., a carboxyaldehyde group), acetyl, trifluoroacetyl, propionyl, and butanoyl.
  • exemplary unsubstituted acyl groups include from 1 to 6, from 1 to 11, or from 1 to 21 carbons.
  • alkyl refers to a branched or straight-chain monovalent saturated aliphatic hydrocarbon radical of 1 to 20 carbon atoms (e.g., 1 to 16 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms).
  • An alkylene is a divalent alkyl group.
  • alkenyl refers to a straight-chain or branched hydrocarbon residue having a carbon-carbon double bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6, or 2 carbon atoms).
  • alkynyl refers to a straight-chain or branched hydrocarbon residue having a carbon-carbon triple bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6, or 2 carbon atoms).
  • amino represents -N(R N1 )2, wherein each R N1 is, independently, H, OH, NO2, N(R N2 )2, SO2OR N2 , SO2R N2 , SOR N2 , an N-protecting group, alkyl, alkoxy, aryl, arylalkyl, cycloalkyl, acyl (e.g., acetyl, trifluoroacetyl, or others described herein), wherein each of these recited R N1 groups can be optionally substituted; or two R N1 combine to form an alkylene or heteroalkylene, and wherein each R N2 is, independently, H, alkyl, or aryl.
  • each R N1 is, independently, H, alkyl, or aryl.
  • the amino groups of the invention can be an unsubstituted amino (i.e., -NH2) or a substituted amino (i.e., -N(R N1 )2).
  • aryl refers to an aromatic mono- or polycarbocyclic radical of 6 to 12 carbon atoms having at least one aromatic ring. Examples of such groups include, but are not limited to, phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, 1,2-dihydronaphthyl, indanyl, and 1H-indenyl.
  • arylalkyl represents an alkyl group substituted with an aryl group.
  • Exemplary unsubstituted arylalkyl groups are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as C1-C6 alkyl C6-10 aryl, C1-C10 alkyl C6-10 aryl, or C1-C20 alkyl C6-10 aryl), such as, benzyl and phenethyl.
  • the akyl and the aryl each can be further substituted with 1, 2, 3, or 4 substituent groups as defined herein for the respective groups.
  • the term “azido,” as used herein, represents a -N3 group.
  • cyano represents a CN group.
  • Carbocyclyl refer to a non-aromatic C3-C12 monocyclic, bicyclic, or tricyclic structure in which the rings are formed by carbon atoms.
  • Carbocyclyl structures include cycloalkyl groups and unsaturated carbocyclyl radicals.
  • cycloalkyl refers to a saturated, non-aromatic, monovalent mono- or polycarbocyclic radical of three to ten, preferably three to six carbon atoms.
  • halo means a fluorine (fluoro), chlorine (chloro), bromine (bromo), or iodine (iodo) radical.
  • heteroalkyl refers to an alkyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur.
  • the heteroalkyl group can be further substituted with 1, 2, 3, or 4 substituent groups as described herein for alkyl groups.
  • heteroalkyl groups are an “alkoxy” which, as used herein, refers alkyl-O- (e.g., methoxy and ethoxy).
  • a heteroalkylene is a divalent heteroalkyl group.
  • heteroalkenyl refers to an alkenyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur.
  • the heteroalkenyl group can be further substituted with 1, 2, 3, or 4 substituent groups as described herein for alkenyl groups.
  • heteroalkenyl groups are an “alkenoxy” which, as used herein, refers alkenyl-O-.
  • a heteroalkenylene is a divalent heteroalkenyl group.
  • heteroalkynyl refers to an alkynyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur.
  • the heteroalkynyl group can be further substituted with 1, 2, 3, or 4 substituent groups as described herein for alkynyl groups.
  • heteroalkynyl groups are an “alkynoxy” which, as used herein, refers alkynyl-O-.
  • heteroalkynylene is a divalent heteroalkynyl group.
  • heteroaryl refers to an aromatic mono- or polycyclic radical of 5 to 12 atoms having at least one aromatic ring containing one, two, or three ring heteroatoms selected from N, O, and S, with the remaining ring atoms being C. One or two ring carbon atoms of the heteroaryl group may be replaced with a carbonyl group. Examples of heteroaryl groups are pyridyl, pyrazoyl, benzooxazolyl, benzoimidazolyl, benzothiazolyl, imidazolyl, oxaxolyl, and thiazolyl.
  • heteroarylalkyl represents an alkyl group substituted with a heteroaryl group.
  • exemplary unsubstituted heteroarylalkyl groups are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as C1-C6 alkyl C2-C9 heteroaryl, C1-C10 alkyl C2-C9 heteroaryl, or C1-C20 alkyl C2-C9 heteroaryl).
  • the akyl and the heteroaryl each can be further substituted with 1, 2, 3, or 4 substituent groups as defined herein for the respective groups.
  • heterocyclyl denotes a mono- or polycyclic radical having 3 to 12 atoms having at least one ring containing one, two, three, or four ring heteroatoms selected from N, O or S and no aromatic ring containing any N, O, or S atoms.
  • heterocyclyl groups include, but are not limited to, morpholinyl, thiomorpholinyl, furyl, piperazinyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydropyranyl, tetrahydrofuranyl, and 1,3-dioxanyl.
  • heterocyclylalkyl represents an alkyl group substituted with a heterocyclyl group.
  • exemplary unsubstituted heterocyclylalkyl groups are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as C1-C6 alkyl C2-C9 heterocyclyl, C1-C10 alkyl C2-C9 heterocyclyl, or C1-C20 alkyl C2-C9 heterocyclyl).
  • the akyl and the heterocyclyl each can be further substituted with 1, 2, 3, or 4 substituent groups as defined herein for the respective groups.
  • hydroxyl represents an -OH group.
  • N-protecting group represents those groups intended to protect an amino group against undesirable reactions during synthetic procedures. Commonly used N-protecting groups are disclosed in Greene, “Protective Groups in Organic Synthesis,” 3 rd Edition (John Wiley & Sons, New York, 1999).
  • N-protecting groups include acyl, aryloyl, or carbamyl groups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, ⁇ -chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, and chiral auxiliaries such as protected or unprotected D, L or D, L-amino acids such as alanine, leucine, and phenylalanine; sulfonyl-containing groups such as benzenesulfonyl, and p-toluenesulfonyl; carbamate forming groups such as benzyloxycarbonyl, p-
  • N-protecting groups are alloc, formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, alanyl, phenylsulfonyl, benzyl, t-butyloxycarbonyl (Boc), and benzyloxycarbonyl (Cbz).
  • nitro represents an NO2 group.
  • thiol represents an -SH group.
  • alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl (e.g., cycloalkyl), aryl, heteroaryl, and heterocyclyl groups may be substituted or unsubstituted. When substituted, there will generally be 1 to 4 substituents present, unless otherwise specified.
  • Substituents include, for example: aryl (e.g., substituted and unsubstituted phenyl), carbocyclyl (e.g., substituted and unsubstituted cycloalkyl), halo (e.g., fluoro), hydroxyl, oxo, heteroalkyl (e.g., substituted and unsubstituted methoxy, ethoxy, or thioalkoxy), heteroaryl, heterocyclyl, amino (e.g., NH2 or mono- or dialkyl amino), azido, cyano, nitro, or thiol.
  • aryl e.g., substituted and unsubstituted phenyl
  • carbocyclyl e.g., substituted and unsubstituted cycloalkyl
  • halo e.g., fluoro
  • hydroxyl oxo
  • heteroalkyl e.g., substituted and
  • Aryl, carbocyclyl (e.g., cycloalkyl), heteroaryl, and heterocyclyl groups may also be substituted with alkyl (unsubstituted and substituted such as arylalkyl (e.g., substituted and unsubstituted benzyl)).
  • Compounds of the invention can have one or more asymmetric carbon atoms and can exist in the form of optically pure enantiomers, mixtures of enantiomers such as, for example, racemates, optically pure diastereoisomers, mixtures of diastereoisomers, diastereoisomeric racemates or mixtures of diastereoisomeric racemates.
  • optically active forms can be obtained for example by resolution of the racemates, by asymmetric synthesis or asymmetric chromatography (chromatography with a chiral adsorbent or eluant). That is, certain of the disclosed compounds may exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement. Enantiomers are pairs of stereoisomers whose mirror images are not superimposable, most commonly because they contain an asymmetrically substituted carbon atom that acts as a chiral center. "Enantiomer” means one of a pair of molecules that are mirror images of each other and are not superimposable.
  • Diastereomers are stereoisomers that are not related as mirror images, most commonly because they contain two or more asymmetrically substituted carbon atoms and represent the configuration of substituents around one or more chiral carbon atoms.
  • Enantiomers of a compound can be prepared, for example, by separating an enantiomer from a racemate using one or more well-known techniques and methods, such as, for example, chiral chromatography and separation methods based thereon. The appropriate technique and/or method for separating an enantiomer of a compound described herein from a racemic mixture can be readily determined by those of skill in the art.
  • Racemate or “racemic mixture” means a compound containing two enantiomers, wherein such mixtures exhibit no optical activity; i.e., they do not rotate the plane of polarized light.
  • “Geometric isomer” means isomers that differ in the orientation of substituent atoms in relationship to a carbon-carbon double bond, to a cycloalkyl ring, or to a bridged bicyclic system. Atoms (other than H) on each side of a carbon- carbon double bond may be in an E (substituents are on opposite sides of the carbon- carbon double bond) or Z (substituents are oriented on the same side) configuration.
  • R,” “S,” “S*,” “R*,” “E,” “Z,” “cis,” and “trans,” indicate configurations relative to the core molecule.
  • Certain of the disclosed compounds may exist in atropisomeric forms.
  • Atropisomers are stereoisomers resulting from hindered rotation about single bonds where the steric strain barrier to rotation is high enough to allow for the isolation of the conformers.
  • the compounds of the invention may be prepared as individual isomers by either isomer-specific synthesis or resolved from an isomeric mixture.
  • Conventional resolution techniques include forming the salt of a free base of each isomer of an isomeric pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming the salt of the acid form of each isomer of an isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), forming an ester or amide of each of the isomers of an isomeric pair using an optically pure acid, amine or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or resolving an isomeric mixture of either a starting material or a final product using various well known chromatographic methods.
  • the stereochemistry of a disclosed compound is named or depicted by structure
  • the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99% or 99.9%) by weight relative to the other stereoisomers.
  • the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by weight optically pure.
  • the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by weight pure.
  • Percent optical purity is the ratio of the weight of the enantiomer or over the weight of the enantiomer plus the weight of its optical isomer. Diastereomeric purity by weight is the ratio of the weight of one diastereomer or over the weight of all the diastereomers.
  • the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by mole fraction pure relative to the other stereoisomers.
  • the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by mole fraction pure.
  • diastereomer When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by mole fraction pure. Percent purity by mole fraction is the ratio of the moles of the enantiomer or over the moles of the enantiomer plus the moles of its optical isomer. Similarly, percent purity by moles fraction is the ratio of the moles of the diastereomer or over the moles of the diastereomer plus the moles of its isomer.
  • the term “a” may be understood to mean “at least one”; (ii) the term “or” may be understood to mean “and/or”; (iii) the terms “comprising” and “including” may be understood to encompass itemized components or steps whether presented by themselves or together with one or more additional components or steps; and (iv) the terms “about” and “approximately” may be understood to permit standard variation as would be understood by those of ordinary skill in the art; and (v) where ranges are provided, endpoints are included.
  • the term “administration” refers to the administration of a composition (e.g., a compound, a complex or a preparation that includes a compound or complex as described herein) to a subject or system.
  • Administration to an animal subject may be by any appropriate route.
  • administration may be bronchial (including by bronchial instillation), buccal, enteral, interdermal, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal and vitreal.
  • the term “animal” refers to any member of the animal kingdom. In some embodiments, “animal” refers to humans, at any stage of development.
  • animal refers to non-human animals, at any stage of development.
  • the non-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, cattle, a primate, and/or a pig).
  • animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, and/or worms.
  • an animal may be a transgenic animal, genetically-engineered animal, and/or a clone.
  • the terms “approximately” and “about” are each intended to encompass normal statistical variation as would be understood by those of ordinary skill in the art as appropriate to the relevant context.
  • the terms “approximately” or “about” each refer to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of a stated value, unless otherwise stated or otherwise evident from the context (e.g., where such number would exceed 100% of a possible value).
  • Two events or entities are “associated” with one another, as that term is used herein, if the presence, level and/or form of one is correlated with that of the other.
  • a particular entity e.g., polypeptide
  • a particular disease, disorder, or condition if its presence, level and/or form correlates with incidence of and/or susceptibility of the disease, disorder, or condition (e.g., across a relevant population).
  • a subject such as a human subject undergoing therapy for the treatment of a neurological disorder, for example, amyotrophic lateral sclerosis, frontotemporal degeneration (also referred to as frontotemporal lobar degeneration and frontotemporal dementia), Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’s disease, Inclusion body myopathy with early-onset Paget disease and frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease, and hereditary inclusion body myopathy.
  • a neurological disorder for example, amyotrophic lateral sclerosis, frontotemporal degeneration (also referred to as frontotemporal lobar degeneration and frontotemporal dementia), Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, cor
  • exemplary benefits in the context of a subject undergoing treatment for a neurological disorder using the compositions and methods described herein include the slowing and halting of disease progression, as well as suppression of one or more symptoms associated with the disease.
  • examples of clinical “benefits” and “responses” are (i) an improvement in the subject’s condition as assessed using the amyotrophic lateral sclerosis functional rating scale (ALSFRS) or the revised ALSFRS (ALSFRS-R) following administration of the CYP51A1 inhibitor, such as an improvement in the subject’s ALSFRS or ALSFRS-R score within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an improvement in the subject’s ALSFRS or ALSFRS-R score within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor
  • an “effective amount” of any one of the compounds of the invention or a combination of any of the compounds of the invention or a pharmaceutically acceptable salt thereof is administered via any of the usual and acceptable methods known in the art, either singly or in combination.
  • the terms “cytochrome P450 isoform 51A1,” “CYP51A1,” and “lanosterol 14- alpha demethylase” are used interchangeably and refer to the enzyme that catalyzes the conversion of lanosterol to 4,4-dimethylcholesta-8(9),14,24-trien-3 ⁇ -ol, for example, in human subjects.
  • cytochrome P450 isoform 51A1 refers not only to wild-type forms of CYP51A1, but also to variants of wild-type CYP51A1 proteins and nucleic acids encoding the same.
  • the amino acid sequence and corresponding mRNA sequence of a wild-type form of human CYP51A1 are provided herein as SEQ ID NOs: 1 and 2, which correspond to GenBank Accession No. AAC50951.1 and NCBI Reference Sequence NO. NM_000786.3, respectively. These sequences are shown in Table 2, below. Table 2. Amino acid and nucleic acid sequences of wild-type human CYP5A1
  • cytochrome P450 isoform 51A1 “CYP51A1,” and “lanosterol 14-alpha demethylase” as used herein include, for example, forms of the human CYP51A1 protein that have an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 1 (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9%, or 100% identical to the amino acid sequence of SEQ ID NO: 1) and/or forms of the human CYP51A1 protein that contain one or more substitutions, insertions, and/or deletions (e.g., one or more conservative and/or nonconservative amino acid substitutions, such as up to 5, 10, 15, 20, 25, or more, conservative or nonconservative amino acid substitutions) relative to a wild-type CYP51A1 protein.
  • substitutions, insertions, and/or deletions
  • cytochrome P450 isoform 51A1 inhibitor As used herein, the terms “cytochrome P450 isoform 51A1 inhibitor,” “CYP51A1 inhibitor,” and “lanosterol 14-alpha demethylase inhibitor” are used interchangeably and refer to substances, such as compounds of Formula I.
  • Inhibitors of this type may, for example, competitively inhibit CYP51A1 activity by specifically binding the CYP51A1 enzyme (e.g., by virtue of the affinity of the inhibitor for the CYP51A1 active site), thereby precluding, hindering, or halting the entry of one or more endogenous substrates of CYP51A1 into the enzyme’s active site.
  • cytochrome P450 isoform 51A1 inhibitor refers to substances that reduce the concentration and/or stability of CYP51A1 mRNA transcripts in vivo, as well as those that suppress the translation of functional CYP51A1 enzyme.
  • CYP51A1-associated disorder refers to an undesired physiological condition, disorder, or disease that is associated with and/or mediated at least in part by CYP51A1. In some instances, CYP51A1-associated disorders are associated with excess CYP51A1 levels and/or activity.
  • CYP51A1-associated disorders include but are not limited to central nervous system (CNS) disorders, dementia, Alzheimer's Disease, chronic traumatic encephalopathy, FTLD-TDP, LATE, or frontotemporal lobar degeneration.
  • CNS central nervous system
  • FTLD-TDP traumatic encephalopathy
  • LATE frontotemporal lobar degeneration
  • combination therapy refers to those situations in which a subject is simultaneously exposed to two or more therapeutic agents.
  • two or more compounds may be administered simultaneously; in some embodiments, such compounds may be administered sequentially; in some embodiments, such compounds are administered in overlapping dosing regimens.
  • the term “dosage form” refers to a physically discrete unit of an active compound (e.g., a therapeutic or diagnostic agent) for administration to a subject.
  • Each unit contains a predetermined quantity of active agent.
  • such quantity is a unit dosage amount (or a whole fraction thereof) appropriate for administration in accordance with a dosing regimen that has been determined to correlate with a desired or beneficial outcome when administered to a relevant population (i.e., with a therapeutic dosing regimen).
  • a dosage amount or a whole fraction thereof
  • a dosing regimen refers to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time.
  • a given therapeutic compound has a recommended dosing regimen, which may involve one or more doses.
  • a dosing regimen comprises a plurality of doses each of which are separated from one another by a time period of the same length; in some embodiments, a dosing regimen comprises a plurality of doses and at least two different time periods separating individual doses.
  • all doses within a dosing regimen are of the same unit dose amount. In some embodiments, different doses within a dosing regimen are of different amounts.
  • a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount different from the first dose amount. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount same as the first dose amount. In some embodiments, a dosing regimen is correlated with a desired or beneficial outcome when administered across a relevant population (i.e., is a therapeutic dosing regimen).
  • the term “neuromuscular disorder” refers to a disease impairing the ability of one or more neurons to control the activity of an associated muscle.
  • neuromuscular disorders are amyotrophic lateral sclerosis, congenital myasthenic syndrome, congenital myopathy, cramp fasciculation syndrome, Duchenne muscular dystrophy, glycogen storage disease type II, hereditary spastic paraplegia, inclusion body myositis, Isaac's Syndrome, Kearns-Sayre syndrome, Lambert–Eaton myasthenic syndrome, mitochondrial myopathy, muscular dystrophy, myasthenia gravis, myotonic dystrophy, peripheral neuropathy, spinal and bulbar muscular atrophy, spinal muscular atrophy, Stiff person syndrome, Troyer syndrome, and Guillain–Barré syndrome, among others.
  • composition represents a composition containing a compound described herein formulated with a pharmaceutically acceptable excipient, and manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for the treatment of disease in a mammal.
  • Pharmaceutical compositions can be formulated, for example, for oral administration in unit dosage form (e.g., a tablet, capsule, caplet, gelcap, or syrup); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate emboli and in a solvent system suitable for intravenous use); or in any other pharmaceutically acceptable formulation.
  • a “pharmaceutically acceptable excipient,” as used herein, refers any ingredient other than the compounds described herein (for example, a vehicle capable of suspending or dissolving the active compound) and having the properties of being substantially nontoxic and non-inflammatory in a patient.
  • Excipients may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspensing or dispersing agents, sweeteners, and waters of hydration.
  • pharmaceutically acceptable salt means any pharmaceutically acceptable salt of the compound of formula (I).
  • pharmaceutically acceptable salts of any of the compounds described herein include those that are within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and are commensurate with a reasonable benefit/risk ratio.
  • Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and in Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008.
  • the salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting a free base group with a suitable organic acid.
  • the compounds of the invention may have ionizable groups so as to be capable of preparation as pharmaceutically acceptable salts.
  • These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds of the invention be prepared from inorganic or organic bases.
  • the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases.
  • Suitable pharmaceutically acceptable acids and bases and methods for preparation of the appropriate salts are well-known in the art. Salts may be prepared from pharmaceutically acceptable non-toxic acids and bases including inorganic and organic acids and bases.
  • pure means substantially pure or free of unwanted components (e.g., other compounds and/or other components of a cell lysate), material defilement, admixture or imperfection.
  • unwanted components e.g., other compounds and/or other components of a cell lysate
  • material defilement e.g., cell lysate
  • admixture e.g., cell lysate
  • imperfection e.g., a variety of clinical indicators can be used to identify a patient as “at risk” of developing a particular neurological disease.
  • Examples of patients that are “at risk” of developing a neurological disease, such as amyotrophic lateral sclerosis, frontotemporal degeneration, Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’s disease, Inclusion body myopathy with early-onset Paget disease and frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease, and hereditary inclusion body myopathy, include (i) subjects exhibiting or prone to exhibit aggregation of TAR-DNA binding protein (TDP)-43, and (ii) subjects expressing a mutant form of TDP-43 containing a mutation associated with TDP-43 aggregation and toxicity, such as a mutation selected from Q331K, M337V, Q
  • Subjects that are “at risk” of developing amyotrophic lateral sclerosis may exhibit one or both of these characteristics, for example, prior to the first administration of a CYP51A1 inhibitor in accordance with the compositions and methods described herein.
  • TAR-DNA binding protein-43 and “TDP-43” are used interchangeably and refer to the transcription repressor protein involved in modulating HIV-1 transcription and alternative splicing of the cystic fibrosis transmembrane conductance regulator (CFTR) pre-mRNA transcript, for example, in human subjects.
  • CFTR cystic fibrosis transmembrane conductance regulator
  • TAR-DNA binding protein-43 and “TDP-43” refer not only to wild-type forms of TDP-43, but also to variants of wild-type TDP-43 proteins and nucleic acids encoding the same.
  • the amino acid sequence and corresponding mRNA sequence of a wild-type form of human TDP-43 are provided herein as SEQ ID NOs: 3 and 4, which correspond to NCBI Reference Sequence NOs. NM_007375.3 and NP_031401.1, respectively. These sequences are shown in Table 3, below. Table 3.
  • TAR-DNA binding protein-43 and “TDP-43” as used herein include, for example, forms of the human TDP-43 protein that have an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 3 (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9%, or 100% identical to the amino acid sequence of SEQ ID NO: 3) and/or forms of the human TDP-43 protein that contain one or more substitutions, insertions, and/or deletions (e.g., one or more conservative and/or nonconservative amino acid substitutions, such as up to 5, 10, 15, 20, 25, or more, conservative or nonconservative amino acid substitutions) relative to a wild- type TDP-43 protein.
  • substitutions, insertions, and/or deletions e.g., one or more conservative and/or nonconservative amino acid substitutions, such as up
  • patients that may be treated for a neurological disorder as described herein include human patients that express a form of TDP-43 having a mutation associated with elevated TDP-43 aggregation and toxicity, such as a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D.
  • a neurological disorder as described herein such as amyotrophic lateral sclerosis, frontotemporal degeneration, Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’s disease, Inclusion body myopathy with early-onset Paget disease and frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease, and hereditary inclusion body myopathy, include human patients that express a form of TDP-43 having
  • TAR-DNA binding protein-43 and “TDP-43” as used herein include, for example, forms of the human TDP-43 gene that encode an mRNA transcript having a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 4 (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9%, or 100% identical to the amino acid sequence of SEQ ID NO: 4).
  • the term “subject” refers to any organism to which a composition in accordance with the invention may be administered, e.g., for experimental, diagnostic, prophylactic, and/or therapeutic purposes. Typical subjects include any animal (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans). A subject may seek or be in need of treatment, require treatment, be receiving treatment, be receiving treatment in the future, or be a human or animal who is under care by a trained professional for a particular disease or condition.
  • animal e.g., mammals such as mice, rats, rabbits, non-human primates, and humans.
  • a subject may seek or be in need of treatment, require treatment, be receiving treatment, be receiving treatment in the future, or be a human or animal who is under care by a trained professional for a particular disease or condition.
  • treat means both therapeutic treatment and prophylactic or preventative measures wherein the object is to prevent or slow down (lessen) an undesired physiological condition, disorder, or disease, or obtain beneficial or desired clinical results.
  • Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of the extent of a condition, disorder, or disease; stabilized (i.e., not worsening) state of condition, disorder, or disease; delay in onset or slowing of condition, disorder, or disease progression; amelioration of the condition, disorder, or disease state or remission (whether partial or total), whether detectable or undetectable; an amelioration of at least one measurable physical parameter, not necessarily discernible by the patient; or enhancement or improvement of condition, disorder, or disease.
  • Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival as compared to expected survival if not receiving treatment.
  • a “therapeutic regimen” refers to a dosing regimen whose administration across a relevant population is correlated with a desired or beneficial therapeutic outcome.
  • the term “therapeutically effective amount” means an amount that is sufficient, when administered to a population suffering from or susceptible to a disease, disorder, and/or condition in accordance with a therapeutic dosing regimen, to treat the disease, disorder, and/or condition.
  • a therapeutically effective amount is one that reduces the incidence and/or severity of, and/or delays onset of, one or more symptoms of the disease, disorder, and/or condition.
  • a therapeutically effective amount does not in fact require successful treatment be achieved in a particular individual.
  • a therapeutically effective amount may be that amount that provides a particular desired pharmacological response in a significant number of subjects when administered to patients in need of such treatment. It is specifically understood that particular subjects may, in fact, be “refractory” to a “therapeutically effective amount.” To give but one example, a refractory subject may have a low bioavailability such that clinical efficacy is not obtainable.
  • reference to a therapeutically effective amount may be a reference to an amount as measured in one or more specific tissues (e.g., a tissue affected by the disease, disorder or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine, etc).
  • a therapeutically effective amount may be formulated and/or administered in a single dose. In some embodiments, a therapeutically effective amount may be formulated and/or administered in a plurality of doses, for example, as part of a dosing regimen.
  • FIGS.1A – 1C demonstrate that the viability of a yeast TDP-43 model is restored by the Erg11 inhibitor, fluconazole.
  • FIG.1A Structure of the Erg11 inhibitor and anti-fungal, fluconazole.
  • FIG.1B Fluconazole rescues viability of TDP-43-expressing yeast using a resazurin-reduction endpoint.
  • FIG. 1C Wild-type yeast cultures were treated with fluconazole for eight hours prior to HPLC analysis for lanosterol and ergosterol. Data are expressed as the area under the curve (AUC) normalized to cell mass based on optical density of cultures at 600 nm. Fluconazole treatment reduces ergosterol, while simultaneously leading to an increase in the Erg11 substrate, lanosterol.
  • FIG.2 shows the structures of compounds used in primary rat cortical neuron TDP-43 wild type and Q331K mutant survival studies.
  • FIGS.3A and 3B demonstrate that compound A promotes survival in primary rat cortical neurons transfected with wild-type TDP-43.
  • Rat primary cortical neurons were co-transfected with a red fluorescent protein (RFP) as a morphological marker and either control (empty vector) or wild-type TDP- 43 expression plasmids and treated with vehicle (DMSO) or a titration of compound A.
  • RFP red fluorescent protein
  • FIGS.3A and 3B Forest plots.
  • FIGS.4A and 4B demonstrate that compound A promotes survival in primary rat cortical neurons transfected with Q331K Mutant TDP-43.
  • Rat primary cortical neurons were co-transfected with a red fluorescent protein (RFP) as a morphological marker and either control (empty vector) or Q331K mutant TDP-43 expression plasmids and treated with vehicle (DMSO) or a titration of compound A.
  • RFP red fluorescent protein
  • DMSO vehicle
  • FIG.4A Risk of neuron death plots. The lifetime of each neuron was determined by either loss of RFP signal or morphological indicators of death such as loss of neurites and cell blebbing and used to generate cumulative hazard plots of risk of death over time (hrs) post-transfection.
  • FIG.4B Forest plots. Hazard ratios for each treatment group (relative to TDP-43 DMSO group) were determined by cox regression analysis and used to generate forest plots.
  • FIGS.5A and 5B demonstrate that compound B promotes survival in primary rat cortical neurons transfected with wild-type TDP-43. Rat primary cortical neurons were co-transfected with a red fluorescent protein (RFP) as a morphological marker and either control (empty vector) or wild type TDP- 43 expression plasmids and treated with vehicle (DMSO) or a titration of compound B.
  • RFP red fluorescent protein
  • FIG.5A Risk of neuron death plots.
  • the lifetime of each neuron was determined by either loss of RFP signal or morphological indicators of death such as loss of neurites and cell blebbing and used to generate cumulative hazard plots of risk of death over time (hrs) post-transfection.
  • FOG.5B Forest plots.
  • Hazard ratios for each treatment group were determined by cox regression analysis and used to generate forest plots.
  • Hazard ratios (HR) ⁇ 1 in which the confidence interval (CI) does not encompass 1 represent treatments that significantly reduce probability of neuron death relative to the TDP-43 DMSO control.
  • P p-value.
  • the present invention features compositions and methods for treating neurological disorders, such as amyotrophic lateral sclerosis and other neuromuscular disorders, as well as frontotemporal degeneration, Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’s disease, Inclusion body myopathy with early-onset Paget disease and frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease, and hereditary inclusion body myopathy among others.
  • neurological disorders such as amyotrophic lateral sclerosis and other neuromuscular disorders, as well as frontotemporal degeneration, Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’
  • the invention provides inhibitors of cytochrome P450 isoform 51A1 (CYP51A1), also referred to herein as lanosterol 14-alpha demethylase, that may be administered to a patient (e.g., a human patient) so as to treat or prevent a neurological disorder, such as one or more of the foregoing conditions.
  • a patient e.g., a human patient
  • the CYP51A1 inhibitor may be administered to the patient to alleviate one or more symptoms of the disorder and/or to remedy an underlying molecular pathology associated with the disease, such as to suppress or prevent aggregation of TAR-DNA binding protein (TDP)-43.
  • TDP TAR-DNA binding protein
  • TDP-43 aggregation modulates TDP- 43 aggregation in vivo. Suppression of TDP-43 aggregation exerts beneficial effects in patients suffering from a neurological disorder.
  • Many pathological conditions have been correlated with TDP-43-promoted aggregation and toxicity, such as amyotrophic lateral sclerosis, frontotemporal degeneration, Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’s disease, IBMPFD, sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease, and hereditary inclusion body myopathy.
  • patients suffering from diseases associated with TDP-43 aggregation and toxicity may be treated, for example, due to the suppression of TDP-43 aggregation induced by the CYP51A1 inhibitor.
  • Patients that are likely to respond to CYP51A1 inhibition as described herein include those that have or are at risk of developing TDP-43 aggregation, such as those that express a mutant form of TDP- 43 associated with TDP-43 aggregation and toxicity in vivo.
  • compositions and methods described herein thus provide the additional clinical benefit of enabling the identification of patients that are likely to respond to CYP51A1 inhibitor therapy, as well as processes for treating these patients accordingly.
  • the sections that follow provide a description of exemplary CYP51A1 inhibitors that may be used in conjunction with the compositions and methods disclosed herein.
  • the sections below additionally provide a description of various exemplary routes of administration and pharmaceutical compositions that may be used for delivery of these substances for the treatment of a neurological disorder.
  • CYP51A1 inhibitors described herein include compounds having a structure according to Formula I: Formula I wherein n is 0, 1, 2, 3, or 4; X is S or O; each R 1 is, independently, halo, cyano, hydroxy, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, -SO2-optionally substituted C1-C6 alkyl, or -CO2-optionally substituted C1-C6 alkyl; R 2 is optionally substituted C2-C9 heteroaryl; and R 3 is optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C9 heterocyclyl, optionally substituted C1-C6 alkyl C3-C8 cycloalkyl, optionally substituted C1-C6 heteroalkyl C3-C8 cycloalkyl, optionally substituted C1-C6 alkyl, optionally substituted C1-C6
  • the compound has the structure of any one of compounds 1-273 in Table 1.
  • Other embodiments, as well as exemplary methods for the synthesis or production of these compounds, are described herein.
  • Methods of Treatment Suppression of CYP51A1 Activity and TDP-43 Aggregation to Treat Neurological Disorders Using the compositions and methods described herein, a patient suffering from a neurological disorder may be administered a CYP51A1 inhibitor, such as a small molecule, antibody, antigen-binding fragment thereof, or interfering RNA molecule described herein, so as to treat the disorder and/or to suppress one or more symptoms associated with the disorder.
  • a CYP51A1 inhibitor such as a small molecule, antibody, antigen-binding fragment thereof, or interfering RNA molecule described herein, so as to treat the disorder and/or to suppress one or more symptoms associated with the disorder.
  • Exemplary neurological disorders that may be treated using the compositions and methods described herein are, without limitation, amyotrophic lateral sclerosis, frontotemporal degeneration, Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’s disease, IBMPFD, sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease, and hereditary inclusion body myopathy, as well as neuromuscular diseases such as congenital myasthenic syndrome, congenital myopathy, cramp fasciculation syndrome, Duchenne muscular dystrophy, glycogen storage disease type II, hereditary spastic paraplegia, inclusion body myositis, Isaac's Syndrome, Kearns-Sayre syndrome, Lambert–Eaton myasthenic syndrome, mitochondrial myopathy, muscular dystrophy, myasthenia
  • the present disclosure is based, in part, on the discovery that CYP51A1 inhibitors, such as the agents described herein, are capable of attenuating TDP-43 aggregation in vivo. TDP-43-promoted aggregation and toxicity have been associated with various neurological diseases.
  • the discovery that CYP51A1 inhibitors modulate TDP-43 aggregation provides an important therapeutic benefit.
  • a CYP51A1 inhibitor such as a CYP51A1 inhibitor described herein, a patient suffering from a neurological disorder or at risk of developing such a condition may be treated in a manner that remedies an underlying molecular etiology of the disease.
  • compositions and methods described herein can be used to treat or prevent such neurological conditions, for example, by suppressing the TDP-43 aggregation that promotes pathology. Additionally, the compositions and methods described herein provide the beneficial feature of enabling the identification and treatment of patients that are likely to respond to CYP51A1 inhibitor therapy.
  • a patient e.g., a human patient suffering from or at risk of developing a neurological disease described herein, such as amyotrophic lateral sclerosis
  • a CYP51A1 inhibitor if the patient is identified as likely to respond to this form of treatment.
  • Patients may be identified as such on the basis, for example, of susceptibility to TDP-43 aggregation.
  • the patient is identified is likely to respond to CYP51A1 inhibitor treatment based on the isoform of TDP-43 expressed by the patient.
  • patients expressing TDP-43 isoforms having a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D, among others are more likely to develop TDP-43-promoted aggregation and toxicity relative to patients that do not express such isoforms of TDP-43.
  • a patient may be identified as likely to respond to CYP51A1 inhibitor therapy on the basis of expressing such an isoform of TDP-43, and may subsequently be administered a CYP51A1 inhibitor so as to treat or prevent one or more neurological disorders, such as one or more of the neurological disorders described herein.
  • a patient having a neurological disorder e.g., a patient at risk of developing TDP-43 aggregation, such as a patient expressing a mutant form of TDP-43 having a mutation associated with elevated TDP-43 aggregation and toxicity, for example, a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D
  • a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D is responding favorably to CYP51A1 inhibition.
  • successful treatment of a patient having a neurological disease with a CYP51A1 inhibitor described herein may be signaled by: (i) an improvement in condition as assessed using the amyotrophic lateral sclerosis functional rating scale (ALSFRS) or the revised ALSFRS (ALSFRS-R), such as an improvement in the patient’s ALSFRS or ALSFRS-R score within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an improvement in the patient’s ALSFRS or ALSFRS-R score within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days,
  • the compounds of the invention can be combined with one or more therapeutic agents.
  • the therapeutic agent can be one that treats or prophylactically treats any neurological disorder described herein.
  • Combination Therapies A compound of the invention can be used alone or in combination with other agents that treat neurological disorders or symptoms associated therewith, or in combination with other types of treatment to treat, prevent, and/or reduce the risk of any neurological disorders.
  • the dosages of one or more of the therapeutic compounds may be reduced from standard dosages when administered alone. For example, doses may be determined empirically from drug combinations and permutations or may be deduced by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6, 2005).
  • compositions for administration to human subjects in a biologically compatible form suitable for administration in vivo.
  • the present invention provides a pharmaceutical composition comprising a compound of the invention in admixture with a suitable diluent, carrier, or excipient.
  • the compounds of the invention may be used in the form of the free base, in the form of salts, solvates, and as prodrugs. All forms are within the scope of the invention.
  • the described compounds or salts, solvates, or prodrugs thereof may be administered to a patient in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art.
  • the compounds of the invention may be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump, or transdermal administration and the pharmaceutical compositions formulated accordingly.
  • Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal, and topical modes of administration. Parenteral administration may be by continuous infusion over a selected period of time.
  • a compound of the invention may be orally administered, for example, with an inert diluent or with an assimilable edible carrier, or it may be enclosed in hard or soft shell gelatin capsules, or it may be compressed into tablets, or it may be incorporated directly with the food of the diet.
  • a compound of the invention may be incorporated with an excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, and wafers.
  • a compound of the invention may also be administered parenterally. Solutions of a compound of the invention can be prepared in water suitably mixed with a surfactant.
  • Dispersions can also be prepared in glycerol, liquid polyethylene glycols, DMSO and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms. Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington’s Pharmaceutical Sciences (2003, 20 th ed.) and in The United States Pharmacopeia: The National Formulary (USP 24 NF19), published in 1999.
  • the pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions.
  • compositions for nasal administration may conveniently be formulated as aerosols, drops, gels, and powders.
  • Aerosol formulations typically include a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually presented in single or multidose quantities in sterile form in a sealed container, which can take the form of a cartridge or refill for use with an atomizing device.
  • the sealed container may be a unitary dispensing device, such as a single dose nasal inhaler or an aerosol dispenser fitted with a metering valve which is intended for disposal after use.
  • the dosage form comprises an aerosol dispenser
  • a propellant which can be a compressed gas, such as compressed air or an organic propellant, such as fluorochlorohydrocarbon.
  • the aerosol dosage forms can also take the form of a pump-atomizer.
  • Compositions suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, where the active ingredient is formulated with a carrier, such as sugar, acacia, tragacanth, gelatin, and glycerine.
  • Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base, such as cocoa butter.
  • the compounds of the invention may be administered to an animal, e.g., a human, alone or in combination with pharmaceutically acceptable carriers, as noted herein, the proportion of which is determined by the solubility and chemical nature of the compound, chosen route of administration, and standard pharmaceutical practice.
  • Dosages The dosage of the compounds of the invention, and/or compositions comprising a compound of the invention, can vary depending on many factors, such as the pharmacodynamic properties of the compound; the mode of administration; the age, health, and weight of the recipient; the nature and extent of the symptoms; the frequency of the treatment, and the type of concurrent treatment, if any; and the clearance rate of the compound in the animal to be treated.
  • One of skill in the art can determine the appropriate dosage based on the above factors.
  • the compounds of the invention may be administered initially in a suitable dosage that may be adjusted as required, depending on the clinical response. In general, satisfactory results may be obtained when the compounds of the invention are administered to a human at a daily dosage of, for example, between 0.05 mg and 3000 mg (measured as the solid form). Dose ranges include, for example, between 10-1000 mg. Alternatively, the dosage amount can be calculated using the body weight of the patient. For example, the dose of a compound, or pharmaceutical composition thereof, administered to a patient may range from 0.1-50 mg/kg.
  • Reaction of amine III with appropriately substituted acyl chloride IV under basic conditions affords amide V.
  • R1, R2, or R3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions.
  • General Scheme 3 Reaction of appropriately substituted amine I with appropriately substituted acid II under a variety of coupling conditions (e.g. HBTU) yields amide III.
  • Reaction of amide III with appropriately substituted alkyl halide IV under basic conditions e.g. potassium carbonate
  • R1, R2, or R3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions.
  • R1, R2, or R3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions.
  • N-protecting groups e.g. trityl
  • General Scheme 7 Condensation of appropriately substituted amine I with appropriately substituted aldehyde II in the presence of a variety of reducing agents (e.g. sodium borohydride) gives appropriately substituted amine III. Coupling of amine III and appropriately substituted amine IV with triphosgene under basic conditions (e.g. triethylamine) yields urea V.
  • R 1 , R 2 , or R 3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions.
  • An appropriately substituted amine I is reacted with appropriately substituted acid II under a variety of coupling conditions (e.g. HATU), or acyl chloride II under basic conditions (e.g. triethylamine), to yield amide or carbamate III.
  • R1, R2, or R3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions.
  • General Scheme 9 Piperidine-4-carboxylic acid I is reacted with appropriately substituted acyl chloride II under basic conditions (e.g. triethylamine) to yield appropriately substituted carboxylic acid III.
  • Carboxylic acid III is coupled with appropriately substituted amine IV under a variety of coupling conditions (e.g.
  • R1, R2, or R3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions.
  • N-protecting groups e.g. trityl
  • Example 1 An appropriately substituted amine I can be reacted with appropriately substituted alcohol II and triphosgene under basic conditions (e.g. triethylamine) to yield carbamate III.
  • R1, R2, or R3 may contain N- protecting groups (e.g. trityl) that require removal under acidic conditions.
  • Example 1
  • Step 1 Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
  • Step 1 Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 2.70 g, 25 mmol
  • 6-ethylbenzo[d]thiazol-2-amine 3.00 g, 17 mmol
  • toluene 60 mL
  • Step 2 Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (1.34 g, 5.0 mmol)
  • 4- cyanobenzoic acid 0.740 g, 5.0 mmol
  • N-N,diisopropylethylamine (1.50 g, 15 mmol) in tetrahydrofuran (50 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (2.80 g, 7.5 mmol).
  • Step 1 Preparation of N-(6-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide
  • Step 1 Preparation of 6-methyl-N-(3-pyridylmethyl)-1,3-benzothiazol-2-amine
  • 3-(chloromethyl)pyridine 0.99 g, 3.0 mmol
  • cesium carbonate 1.98 g, 6.1 mmol
  • Step 2 Preparation of N-(6-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide
  • dichloromethane 5 mL
  • triethylamine 238 mg, 2.4 mmol
  • cyclohexanecarbonyl chloride 86.0 mg, 0.59 mmol
  • Example 3 Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide Step 1: Preparation of 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine To a solution of nicotinaldehyde (1.19 g, 11 mmol), 6-methoxybenzo[d]thiazol-2-amine (2.00 g, 11 mmol) in toluene (10 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide
  • 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 150 mg, 0.55 mmol
  • N,N-diisopropylethylamine 214 mg, 1.7 mmol
  • dichloromethane 10 mL
  • cyclohexanecarbonyl chloride 89.0 mg, 0.61 mmol
  • the residue was purified by prep-HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5 ⁇ m 4.6 ⁇ 50mm column.
  • the elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(6-methoxybenzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)cyclohexanecarboxamide (39.8 mg, 0.10 mmol, 19 %) as a light gray solid.
  • the product was indicated present via UPLC analysis.
  • the crude residue was purified by prep-HPLC (Waters X bridge 150*255uM column; 35-70 % acetonitrile in a 10 mM ammonium acetate solution in water, 12 min gradient).
  • Compound 4-cyano-N-(5- methoxy-1,3-benzothiazol-2-yl)benzamide (320 mg, 1.0 mmol, 60 %) was obtained as a pale yellow solid.
  • Step 1 Preparation of N-benzyl-4-cyano-N-(5-methoxy-1,3-benzothiazol-2-yl)benzamide
  • Step 1 Preparation of N-benzyl-5-methoxybenzo[d]thiazol-2-amine
  • benzaldehyde 0.589 g, 5.6 mmol
  • 5-methoxybenzo[d]thiazol-2-amine (1.00 g, 5.6 mmol) in toluene (10 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • a solution of sodium borohydride (1.06 g, 28 mmol) in ethanol (20 mL) was added.
  • Step 2 Preparation of N-benzyl-4-cyano-N-(5-methoxy-1,3-benzothiazol-2-yl)benzamide
  • thionyl chloride 10 mL
  • Step 2 Preparation of N-benzyl-4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)benzamide
  • N-benzyl-6-ethylbenzo[d]thiazol-2-amine 268 mg, 1.0 mmol
  • 4-cyanobenzoic acid 147 mg, 1.0 mmol
  • triethylamine 303 mg, 3.0 mmol
  • dichloromethane 5 mL
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide 570 mg, 1.5 mmol
  • Step 1 Preparation of 4-cyano-N-(5-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
  • Step 1 Preparation of 5-methoxy-N-(3-pyridylmethyl)-1,3-benzothiazol-2-amine
  • 3-(chloromethyl)pyridine 683 mg, 4.2 mmol
  • potassium carbonate 7.67 mg, 5.6 mmol
  • Step 2 Preparation of 4-cyano-N-(5-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • 5-methoxy-N-(3-pyridylmethyl)-1,3-benzothiazol-2-amine 180 mg, 0.66 mmol
  • dichloromethane 3 mL
  • 4-cyanobenzoyl chloride 165 mg, 1.0 mmol
  • Step 1 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)-2,3-dihydro-1,4- benzodioxine-6-carboxamide
  • Step 1 Preparation of 2,3-dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride
  • 2,3-dihydrobenzo[b][1,4]dioxine-6-carboxylic acid 900 mg, 5.0 mmol
  • thionyl chloride 10 mL
  • Step 2 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)-2,3-dihydro-1,4- benzodioxine-6-carboxamide
  • 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (259 mg, 1.0 mmol) and triethylamine (303 mg, 3.0 mmol) in dichloromethane (5 mL) was added 2,3-dihydrobenzo[b][1,4]dioxine- 6-carbonyl chloride (398 mg, 2.0 mmol) at 0 °C under argon.
  • the mixture was stirred at 40 °C for 4 hours.
  • the product was indicated present via UPLC analysis. Water (30 mL) was added, and the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated.
  • the crude product was purified by Prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10 ⁇ m column.
  • the mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-(4-fluorobenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (12.0 mg, 0.027 mmol, 2.7 %) as a white solid.
  • Step 1 Preparation of N-(4-chloro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3- yl)methyl]propenamide
  • Step 1 Preparation of N-benzyl-5-methoxybenzo[d]thiazol-2-amine
  • 4-chlorobenzo[d]thiazol-2-amine (1.02 g, 5.6 mmol) in toluene (10 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(4-chloro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3-yl)methyl]propanamide
  • 4-chloro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (226 mg, 0.82 mmol)
  • 3- phenoxypropanoic acid 150 mg, 0.90 mmol
  • N,N-diisopropylethylamine (318 mg, 2.5 mmol) in tetrahydrofuran (20 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (468 mg, 1.2 mmol).
  • Example 14 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide Step 1: Preparation of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine To a solution of nicotinaldehyde (1.27 g, 12 mmol) and 4-fluorobenzo[d]thiazol-2-amine (2.00 g, 12 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide
  • 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (259 mg, 1.0 mmol) and 4- (dimethylamino)pyridine (183 mg, 1.5 mmol) in pyridine (5 mL) was added cyclohexanecarbonyl chloride (221 mg, 1.5 mmol) at 0 °C under argon.
  • the mixture was stirred at 40 °C for 12 hours.
  • the product was indicated present via UPLC analysis.
  • Step 1 Preparation of N-[(4-cyanophenyl)methyl]-N-(6-ethyl-1,3-benzothiazol-2-yl)pyridine-3- carboxamide
  • Step 1 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)pyridine-3-carboxamide
  • 6-ethyl-1,3-benzothiazol-2-amine 100 mg, 0.56 mmol
  • pyridine-3-carboxylic acid 7.
  • N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide 214 mg, 0.56 mmol
  • N,N- diisopropylethylamine 3 mL
  • Step 2 Preparation of N-[(4-cyanophenyl)methyl]-N-(6-ethyl-1,3-benzothiazol-2-yl)pyridine-3- carboxamide
  • N-(6-ethyl-1,3-benzothiazol-2-yl)pyridine-3-carboxamide 91.0 mg, 0.32 mmol
  • dimethylformamide (2 mL) stirring.4-(bromomethyl)benzonitrile (62.9 mg, 0.32 mmol) and dipotassium carbonate (53.2 mg, 0.39 mmol) were added and the reaction was stirred at 65°C overnight.
  • the product was indicated via UPLC analysis.
  • Step 1 Preparation of N-(1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
  • Step 1 Preparation of N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde (2.14 g, 20 mmol)
  • benzo[d]thiazol-2-amine (3.00 g, 20 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves.
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
  • N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (241 mg, 1.0 mmol) and 4- (dimethylamino)pyridine (183 mg, 1.5 mmol) in pyridine
  • benzoyl chloride 210 mg, 1.5 mmol
  • the product was indicated present via UPLC analysis. Water (30 mL) was added to the reaction, then the mixture was extracted with dichloromethane (30 mL x 2).
  • Step 1 Preparation of N-(1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide
  • Step 1 Preparation of N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde benzo[d]thiazol-2-amine 3.00 g, 20 mmol
  • toluene 50 mL
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide
  • N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 241 mg, 1.0 mmol
  • 4- (dimethylamino)pyridine 183 mg, 1.5 mmol
  • cyclohexanecarbonyl chloride 220 mg, 1.5 mmol
  • the product was indicated present via UPLC analysis.
  • Example 18 Preparation of 4-cyano-N-(4-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)benzamide Step 1: Preparation of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine To a solution of nicotinaldehyde (1.27 g, 12 mmol) and 4-fluorobenzo[d]thiazol-2-amine (2.00 g, 12 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of 4-cyano-N-(4-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
  • 4-cyanobenzoic acid 147 mg, 1.0 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol)
  • N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide 5 mL
  • 4-fluoro- N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 259 mg, 1.0 mmol).
  • the mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give 4-cyano-N-(4- fluorobenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (14.0 mg, 0.035 mmol, 3.5 %) as a white solid.
  • Step 1 Preparation of N-(1,3-benzothiazol-2-yl)-N-benzyl-4-cyanobenzamide
  • Step 1 Preparation of N-benzylbenzo[d]thiazol-2-amine
  • benzaldehyde (1.06 g, 10 mmol)
  • benzo[d]thiazol-2-amine (1.50 g, 10 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves.
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • a solution of sodium borohydride (1.90 g, 50 mmol) in ethanol (100 mL).
  • the reaction was stirred at 110 °C for 2 hours.
  • Step 2 Preparation of N-(1,3-benzothiazol-2-yl)-N-benzyl-4-cyanobenzamide
  • 4-cyanobenzoic acid 147 mg, 1.0 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol)
  • N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide 5 mL
  • N- benzylbenzo[d]thiazol-2-amine 240 mg, 1.0 mmol
  • Step 1 Preparation of N-(1,3-benzothiazol-2-yl)-4-cyano-N-(pyridin-3-ylmethyl)benzamide
  • Step 1 Preparation of N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde (2.14 g, 20 mmol)
  • benzo[d]thiazol-2-amine (3.00 g, 20 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves.
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • a solution of sodium borohydride (1.90 g, 50 mmol) in ethanol (100 mL).
  • Step 2 Preparation of N-(1,3-benzothiazol-2-yl)-4-cyano-N-(pyridin-3-ylmethyl)benzamide
  • 4-cyanobenzoic acid 147 mg, 1.0 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol)
  • N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide 5 mL
  • N- (pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 0.241 g, 1.00 mmol
  • the mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-(benzo[d]thiazol- 2-yl)-4-cyano-N-(pyridin-3-ylmethyl)benzamide (32.0 mg, 0.085 mmol, 8.5 %) as a white solid.
  • Step 1 Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
  • Step 1 Preparation of 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 1.00 g, 11 mmol
  • 6-methoxybenzo[d]thiazol-2-amine (2.00 g, 11 mmol) in toluene (10 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
  • 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (271 mg, 1.0 mmol) and 4-(dimethylamino)pyridine (183 mg, 1.5 mmol) in pyridine (5 mL) was added benzoyl chloride (210 mg, 1.5 mmol) at 0 °C under argon. The mixture was stirred at 40 °C for 12 hours. The product was indicated present via UPLC analysis.
  • Step 1 Preparation of N-benzyl-4-cyano-N-(6-methoxy-1,3-benzothiazol-2-yl)benzamide
  • Step 1 Preparation of N-benzyl-6-methoxybenzo[d]thiazol-2-amine
  • benzaldehyde 0.590 g, 5.6 mmol
  • 6-methoxybenzo[d]thiazol-2-amine (1.00 g, 5.6 mmol) in toluene (10 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.06 g, 28 mmol) in ethanol (100 mL) was added.
  • Step 2 Preparation of N-benzyl-4-cyano-N-(6-methoxy-1,3-benzothiazol-2-yl)benzamide
  • 4-cyanobenzoic acid 147 mg, 1.0 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol)
  • N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide 5 mL
  • N- benzyl-6-methoxybenzo[d]thiazol-2-amine 270 mg, 1.0 mmol
  • Example 25 Preparation of 4-cyano-N-(6-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)benzamide Step 1: Preparation of 6-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine To a solution of nicotinaldehyde (1.07 g, 10 mmol), 6-methylbenzo[d]thiazol-2-amine (1.64 g, 10 mmol) in toluene (10 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of 4-cyano-N-(6-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
  • 6-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (0.255 g, 1.0 mmol)
  • 4- cyanobenzoic acid (0.735 g, 5.0 mmol)
  • N,N- diisopropylethylamine (1.29 g, 10 mmol) and N,N-dimethylformamide (10 mL) was stirred at room temperature for 1 hour.
  • Step 1 Preparation of 4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)benzamide
  • Step 1 Preparation of 6-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 0.56 g, 8.9 mmol
  • 6-fluorobenzo[d]thiazol-2-amine (1.50 g, 8.9 mmol) in toluene (10 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of 4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
  • thionyl chloride 10 mL
  • Crude product was purified by silica gel column using a gradient of 0-50% ethyl acetate in petroleum ether, then recrystallized by ethanol to offer 4-cyano-N-(6-fluorobenzo[d]thiazol-2- yl)-N-(pyridin-3-ylmethyl)benzamide (63.3 mg, 0.16 mmol, 17 %) as a white solid.
  • Step 2 Preparation of N-(6-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
  • 6-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 255 mg, 1.0 mmol
  • 4-(dimethylamino)pyridine 255 mg, 0.10 mmol
  • benzoyl chloride 154 mg, 1.1 mmol
  • the product was indicated present via UPLC analysis.
  • the product was indicated present via UPLC analysis.
  • the solvent was removed under the reduced pressure.
  • the residue was purified by silica gel column using a gradient of 0-50% ethyl acetate in petroleum ether, then recrystallized by ethanol to offer N-(6-fluorobenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (113 mg, 0.31 mmol, 40 %) as a white solid.
  • Step 1 Preparation of N-(5-methoxy-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
  • Step 1 Preparation of 5-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 1.00 g, 11 mmol
  • 5-methoxybenzo[d]thiazol-2-amine (2.00 g, 11 mmol) in toluene (10 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(5-methoxy-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
  • 5-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 250 mg, 0.92 mmol
  • diisopropylethylamine 357 mg, 2.8 mmol
  • dichloromethane 10 mL
  • benzoyl chloride 142 mg, 1.0 mmol
  • the residue was purified by HPLC (SunFire C18, 4.6*50 mm, 3.5 um column Xbridge C183.5 ⁇ m 4.6 ⁇ 50 mm column.
  • the elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(6-fluorobenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)benzamide (40.8 mg, 0.11 mmol, 12 %) as a white solid.
  • Step 1 Preparation of N-(4-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
  • Step 1 Preparation of 4-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • 4-methylbenzo[d]thiazol-2-amine 3.28 g, 20 mmol
  • toluene 60 mL
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(4-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
  • 4-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 255 mg, 1.0 mmol
  • 4-(dimethylamino)pyridine 12.0 mg, 0.10 mmol
  • benzoyl chloride 154 mg, 1.1 mmol
  • Step 1 Preparation of N-benzyl-4-cyano-N-(6-methyl-1,3-benzothiazol-2-yl)benzamide
  • Step 1 Preparation of N-benzyl-6-methylbenzo[d]thiazol-2-amine
  • benzaldehyde (1.06 g, 10 mmol)
  • 6-methylbenzo[d]thiazol-2-amine (1.64 g, 10 mmol) in toluene (30 mL) was added dried 4 ⁇ molecular sieves.
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • a solution of sodium borohydride (1.90 g, 50 mmol) in ethanol (10 mL) was added.
  • Step 2 Preparation of N-benzyl-4-cyano-N-(6-methyl-1,3-benzothiazol-2-yl)benzamide
  • N-benzyl-6-methylbenzo[d]thiazol-2-amine 0.254 g, 1.0 mmol
  • 4-cyanobenzoic acid 0.735 g, 5.0 mmol
  • N,N-diisopropylethylamine (1.29 g, 10 mmol
  • N,N-dimethylformamide 8 mL
  • Step 1 Preparation of N-benzyl-4-cyano-N-(4-methyl-1,3-benzothiazol-2-yl)benzamide
  • Step 1 Preparation of N-benzyl-4-methylbenzo[d]thiazol-2-amine
  • 4-methylbenzo[d]thiazol-2-amine 3.28 g, 20 mmol
  • benzaldehyde (2.12 g, 20 mmol) in toluene (30 mL) was added dried 4 ⁇ molecular sieves.
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • a solution of sodium borohydride (3.80 g, 100 mmol) in ethanol (20 mL) was added.
  • Step 2 Preparation of N-benzyl-4-cyano-N-(4-methyl-1,3-benzothiazol-2-yl)benzamide
  • N-benzyl-4-methylbenzo[d]thiazol-2-amine (254 mg, 1.0 mmol)
  • 4-cyanobenzoic acid 147 mg, 1.0 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide 570 mg, 1.5 mmol
  • N,N-diisopropylethylamine (645 mg, 5.0 mmol)
  • N,N-dimethylformamide 6 mL
  • Crude product was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5 ⁇ m 4.6 ⁇ 50mm column.
  • the elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(5-methoxybenzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (139 mg, 0.32 mmol, 35 %) as a white solid.
  • Example 34 Preparation of N-(5-methoxy-1,3-benzothiazol-2-yl)-3-phenoxy-N-(pyridin-3- ylmethyl)propenamide
  • thionyl chloride 10 mL
  • Step 1 Preparation of 4-cyano-N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
  • Step 1 Preparation of 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde (1.19 g, 11 mmol)
  • 6-methoxybenzo[d]thiazol-2-amine (2.00 g, 11 mmol) were refluxed with 4 ⁇ molecular sieves in toluene (50 mL) under N2 overnight.
  • Step 2 Preparation of 4-cyano-N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • 4-cyanobenzoic acid 147 mg, 1.0 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol)
  • N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide 5 mL
  • 6- methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 270 mg, 1.0 mmol
  • the mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give 4-cyano-N- (6-methoxybenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (41.0 mg, 0.10 mmol, 10 %) as a white solid.
  • Example 36 Preparation of 4-cyano-N-(4-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
  • Step 1 Preparation of 4-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • 4-methylbenzo[d]thiazol-2-amine 3.28 g, 20 mmol
  • toluene 60 mL
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of 4-cyano-N-(4-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • 4-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (0.255 g, 1.0 mmol)
  • 4- cyanobenzoic acid (0.735 g, 5.0 mmol)
  • N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (2.85 g, 7.5 mmol)
  • N,N- diisopropylethylamine (1.29 g, 10 mmol)
  • N,N-dimethylformamide 10 mL
  • Step 1 Preparation of 4-cyano-N-(4-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
  • Step 1 Preparation of 4-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • 4-methoxybenzo[d]thiazol-2-amine (2.00 g, 11 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of 4-cyano-N-(4-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • 4-cyanobenzoic acid 147 mg, 1.0 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol)
  • N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide 5 mL
  • 4- methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 270 mg, 1.0 mmol
  • the mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give 4-cyano-N-(4- methoxybenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (105 mg, 0.26 mmol, 26 %) as a white solid.
  • Step 1 Preparation of N-(4-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • Step 1 Preparation of 4-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • 4-methoxybenzo[d]thiazol-2-amine (2.00 g, 11 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(4-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • 4-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 270 mg, 1.0 mmol
  • 4-(dimethylamino)pyridine 183 mg, 1.5 mmol
  • benzoyl chloride 210 mg, 1.5 mmol
  • the product was indicated present via UPLC analysis.
  • Step 1 Preparation of N-benzyl-4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)benzamide
  • Step 1 Preparation of N-benzyl-6-fluorobenzo[d]thiazol-2-amine
  • benzaldehyde 0.631 g, 6.0 mmol
  • 6-fluorobenzo[d]thiazol-2-amine (1.00 g, 6.0 mmol) in toluene (10 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.13 g, 30 mmol) in ethanol (20 mL) was added.
  • Step 2 Preparation of N-benzyl-4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)benzamide To a solution of 4-cyanobenzoic acid (125 mg, 0.85 mmol) in dichloromethane (2 mL) was added thionyl chloride (10 mL). The mixture was heated to 100 °C and stirred for 2 hours. The solvent was removed under the reduced pressure.
  • Step 1 Preparation of N-benzyl-4-cyano-N-(4-methoxy-1,3-benzothiazol-2-yl)benzamide
  • Step 1 Preparation of N-benzyl-4-methoxybenzo[d]thiazol-2-amine
  • 4-methoxybenzo[d]thiazol-2-amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.06 g, 28 mmol) in ethanol (100 mL) was added.
  • Step 2 Preparation of N-benzyl-4-cyano-N-(4-methoxy-1,3-benzothiazol-2-yl)benzamide
  • 4-cyanobenzoic acid 147 mg, 1.0 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol)
  • N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide 5 mL
  • N- benzyl-4-methoxybenzo[d]thiazol-2-amine 270 mg, 1.0 mmol
  • the mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-benzyl-4-cyano- N-(4-methoxybenzo[d]thiazol-2-yl)benzamide (26.0 mg, 0.065 mmol, 6.5 %) as a white solid.
  • Step 1 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3- yl)methyl]propenamide
  • Step 1 Preparation of 3-phenoxypropanoyl chloride A mixture of 3-phenoxypropanoic acid (830 mg, 5.0 mmol) in thionyl chloride (10 mL) was stirred at 50 °C for 2 hours, then concentrated under reduced pressure. The resulting solid was 3- phenoxypropanoyl chloride (830 mg, 4.5 mmol, 90 %) as an off-white powder; LCMS (ESI) m/z: 181.1 [M+H] + .
  • Step 2 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3-yl)methyl]propanamide
  • 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (259 mg, 1.0 mmol) and N,N-diisopropylethylamine (387 mg, 3.0 mmol) in dichloromethane (5 mL) was added 3- phenoxypropanoyl chloride (370 mg, 2.0 mmol). The mixture was stirred at 0 °C for 2 hours. The product was indicated present via UPLC analysis.
  • Step 1 Preparation of 4-cyano-N-(5-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
  • Step 1 Preparation of 5-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 0.46 g, 8.8 mmol
  • 5-methylbenzo[d]thiazol-2-amine (1.45 g, 8.8 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of 4-cyano-N-(5-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • 4-cyanobenzoic acid 147 mg, 1.0 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol)
  • N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide 5 mL
  • 5- methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 255 mg, 1.0 mmol.
  • the mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give 4-cyano-N-(5- methylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (77.0 mg, 0.20 mmol, 20 %) as a white solid.
  • Step 1 Preparation of N-(5-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • Step 1 Preparation of 5-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 0.46 g, 8.8 mmol
  • 5-methylbenzo[d]thiazol-2-amine (1.45 g, 8.8 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(5-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • 5-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 255 mg, 1.0 mmol
  • 4-(dimethylamino)pyridine 183 mg, 1.5 mmol
  • benzoyl chloride 210 mg, 1.5 mmol
  • the product was indicated present via UPLC analysis.
  • Step 1 Preparation of N-(5-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 5-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 0.46 g, 8.8 mmol
  • 5-methylbenzo[d]thiazol-2-amine (1.45 g, 8.8 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(5-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • 5-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 255 mg, 1.0 mmol
  • 4-(dimethylamino)pyridine 183 mg, 1.5 mmol
  • cyclohexanecarbonyl chloride 220 mg, 1.5 mmol
  • the product was indicated present via UPLC analysis.
  • Example 45 Preparation of N-benzyl-4-cyano-N-(5-methyl-1,3-benzothiazol-2-yl)benzamide Step 1: Preparation of N-benzyl-5-methylbenzo[d]thiazol-2-amine To a solution of benzaldehyde (290 mg, 2.7 mmol), 5-methylbenzo[d]thiazol-2-amine (450 mg, 2.7 mmol) in toluene (20 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (520 mg, 14 mmol) in ethanol (40 mL) was added.
  • sodium borohydride 520 mg, 14 mmol
  • Step 2 Preparation of N-benzyl-4-cyano-N-(5-methyl-1,3-benzothiazol-2-yl)benzamide
  • 4-cyanobenzoic acid 147 mg, 1.0 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol)
  • N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide 5 mL
  • N- benzyl-5-methylbenzo[d]thiazol-2-amine 254 mg, 1.0 mmol
  • the product was indicated present via UPLC analysis.
  • the solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5 ⁇ m 4.6 ⁇ 50mm column.
  • the elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer 4-cyano-N-((5,6-dihydro-4H-pyrrolo[1,2- b]pyrazol-3-yl)methyl)-N-(6-ethylbenzo[d]thiazol-2-yl)benzamide (30.1 mg, 0.070 mmol, 18 %) as a white solid.
  • the elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer 4-cyano-N-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)methyl)-N-(5- methoxybenzo[d]thiazol-2-yl)benzamide (33.5 mg, 0.080 mmol, 20 %) as a white solid.
  • Step 1 Preparation of N-(5-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • Step 1 Preparation of 5-ethylbenzo[d]thiazol-2-amine
  • a mixture of 3-ethylaniline (5.00 g, 41 mmol) and potassium thiocyanate (4.00 g, 41 mmol) in glacial acetic acid (100 mL) was cooled in an ice bath and stirred for 20 min, and then bromine (6.50 g, 41 mmol) in glacial acetic acid (10 mL) was added dropwise at such a rate to keep the temperature below 10 °C throughout the addition.
  • Step 2 Preparation of 5-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • a mixture of 5-ethylbenzo[d]thiazol-2-amine (2.00 g, 11 mmol), nicotinaldehyde (1.80 g, 17 mmol) and 4 ⁇ molecular sieves in toluene (20 mL) was stirred at 100 °C for 16 hours. The reaction was cooled to room temperature. Then, a solution of sodium borohydride (0.851 mg, 22 mmol) in methanol (5 mL) was added. The suspension was stirred at 60 °C for 2 hours. The product was indicated present via UPLC analysis.
  • Step 3 Preparation of N-(5-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • 5-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 105 mg, 0.39 mmol
  • triethylamine 78.0 mg, 0.78 mmol
  • benzoyl chloride 82.0 mg, 0.59 mmol
  • Step 1 Preparation of N-benzyl-4-cyano-N-(5-ethyl-1,3-benzothiazol-2-yl)benzamide
  • Step 1 Preparation of 4-cyano-N-(5-ethylbenzo[d]thiazol-2-yl)benzamide
  • 5-ethylbenzo[d]thiazol-2-amine 0.356 g, 2.0 mmol
  • 4-cyanobenzoic acid 0.294 g, 2.0 mmol
  • triethylamine 0.606 g, 6.0 mmol
  • dichloromethane 10 mL
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (1.14 g, 3.0 mmol).
  • Step 2 Preparation of N-benzyl-4-cyano-N-(5-ethyl-1,3-benzothiazol-2-yl)benzamide
  • 4-cyano-N-(5-ethylbenzo[d]thiazol-2-yl)benzamide 153 mg, 0.50 mmol
  • (bromomethyl)benzene 170 mg, 1.0 mmol
  • potassium carbonate 138 mg, 1.00 mmol
  • the product was indicated present via UPLC analysis. Water (20 mL) was added, the mixture was extracted with ethyl acetate (3x 20 mL).
  • Step 1 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3- yl)methyl]propenamide
  • Step 1 Preparation of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 481 mg, 4.5 mmol
  • 5-fluorobenzo[d]thiazol-2-amine 500 mg, 3.0 mmol
  • toluene 20 mL
  • Step 2 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3-yl)methyl]propanamide
  • 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 100 mg, 0.39 mmol
  • N,N-diisopropylethylamine 100 mg, 0.78 mmol
  • Example 59 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]thiophene-2- carboxamide Step 1: Preparation of 6-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine To a solution of nicotinaldehyde (0.956 g, 8.9 mmol), 6-fluorobenzo[d]thiazol-2-amine (1.50 g, 8.9 mmol) in toluene (10 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]thiophene-2-carboxamide
  • 6-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 155 mg, 0.60 mmol
  • N,N- diisopropylethylamine 232 mg, 1.8 mmol
  • dichloromethane 5 mL
  • thiophene-2-carbonyl chloride 105 mg, 0.72 mmol
  • Example 60 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]thiophene-2- carboxamide Step 1: Preparation of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine To a solution of nicotinaldehyde (1.27 g, 12 mmol) and 4-fluorobenzo[d]thiazol-2-amine (2.00 g, 12 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]thiophene-2-carboxamide
  • sodium hydride 61.6 mg, 1.5 mmol
  • thiophene-2-carbonyl chloride 169 mg, 1.2 mmol
  • the mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give the desired product N-(4-fluorobenzo[d]thiazol-2- yl)-N-(pyridin-3-ylmethyl)thiophene-2-carboxamide as a white solid (76.4 mg, 0.21 mmol, 27 %).
  • Step 1 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • Step 1 Preparation of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde (1.27 g, 12 mmol)
  • 4-fluorobenzo[d]thiazol-2-amine 2.00 g, 12 mmol
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • sodium hydride 61.6 mg, 1.5 mmol
  • benzoyl chloride 161 mg, 1.2 mmol
  • Step 1 Preparation of N-(1,3-benzothiazol-2-yl)-N-[(pyridin-4- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of N-(pyridin-4-ylmethyl)benzo[d]thiazol-2-amine
  • benzo[d]thiazol-2-amine 0.800 g, 5.8 mmol
  • isonicotinaldehyde 0.621 g, 5.8 mmol
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(1,3-benzothiazol-2-yl)-N-[(pyridin-4-yl)methyl]cyclohexanecarboxamide
  • sodium hydride 66.4 mg, 1.7 mmol
  • cyclohexanecarbonyl chloride 181 mg, 1.2 mmol
  • the product was indicated present via UPLC analysis.
  • the reaction was quenched with crushed ice, then extracted with ethyl acetate (2x 10 mL). The organic layer was washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10 ⁇ m column.
  • the mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give the desired product N-(benzo[d]thiazol-2-yl)-N- (pyridin-4-ylmethyl)cyclohexanecarboxamide as a white solid (82.5 mg, 0.23 mmol, 28 %).
  • Example 64 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]benzamide
  • Step 1 Preparation of N-(6-fluorobenzo[d]thiazol-2-yl)-1-trityl-1H-imidazole-4-carboxamide
  • 6-fluorobenzo[d]thiazol-2-amine 1.68 g, 10 mmol
  • 1-trityl-1H-imidazole-4- carbaldehyde (3.38 g, 10 mmol) in toluene (30 mL) was added dried 4 ⁇ molecular sieves.
  • Step 2 Preparation of N-benzoyl-N-(6-fluorobenzo[d]thiazol-2-yl)-1-trityl-1H-imidazole-4-carboxamide
  • N,N-diisopropylethylamine 310 mg, 2.4 mmol
  • dichloromethane 5 mL
  • benzoyl chloride 134 mg, 0.96 mmol
  • Step 3 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]benzamide
  • N-benzoyl-N-(6-fluorobenzo[d]thiazol-2-yl)-1-trityl-1H-imidazole-4-carboxamide 210 mg, 0.35 mmol
  • dichloromethane 3.5 mL
  • trifluoroacetic acid 1.8 mL
  • the obtained solid was purified by pre-HPLC to offer N-benzoyl-N-(6-fluorobenzo[d]thiazol-2-yl)-1-trityl-1H-imidazole-4-carboxamide (50.4 mg, 0.14 mmol, 41 %) as a white solid.
  • Step 1 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of N-(6-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide
  • N,N-diisopropylethylamine 348 mg, 2.7 mmol
  • dichloromethane 5 mL
  • Step 2 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
  • N-(6-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide 330 mg, 0.55 mmol
  • dichloromethane 5 mL
  • trifluoroacetic acid 2.5 mL
  • Example 66 Preparation of 4-cyano-N-(4,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
  • Step 1 Preparation of 4,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde (1.07 g, 10 mmol
  • 4,6-difluorobenzo[d]thiazol-2-amine (1.86 g, 10 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of 4-cyano-N-(4,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • 4-cyanobenzoic acid 116 mg, 0.79 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide 360 mg, 0.95 mmol
  • N,N-diisopropylethylamine 153 mg, 1.2 mmol
  • N,N-dimethylformamide 5 mL
  • the mobile phase was acetonitrile/0.01% aqueous formic acid) to give 4- cyano-N-(4,6-difluorobenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (69.0 mg, 0.17 mmol, 21 %) as a white solid.
  • Step 1 Preparation of N-(4,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 4,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde (1.07 g, 10 mmol)
  • 4,6-difluorobenzo[d]thiazol-2-amine (1.86 g, 10 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(4,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • 4-(dimethylamino)pyridine 170 mg, 1.2 mmol
  • pyridine 5 mL
  • cyclohexanecarbonyl chloride 140 mg, 1.2 mmol
  • the product was indicated present via UPLC analysis. Water (30 mL) was added, the mixture was extracted with dichloromethane (2x 30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10 ⁇ m column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give N-(4,6-difluorobenzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)cyclohexanecarboxamide (104 mg, 0.27 mmol, 34 %) as a white solid.
  • the elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(4-chlorobenzo[d]thiazol-2-yl)-3-(phenylsulfonyl)-N-(pyridin-3- ylmethyl)propanamide (81.0 mg, 0.17 mmol, 12 %) as a white solid.
  • Step 1 Preparation of N-(1,3-benzothiazol-2-yl)-N-[(pyridin-2- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of N-(pyridin-2-ylmethyl)benzo[d]thiazol-2-amine
  • benzo[d]thiazol-2-amine 0.800 g, 5.8 mmol
  • picolinaldehyde 0.21 g, 5.8 mmol
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(1,3-benzothiazol-2-yl)-N-[(pyridin-2-yl)methyl]cyclohexanecarboxamide
  • sodium hydride 49.6 mg, 1.2 mmol
  • the mixture was stirred at 0 °C for 10 minutes.
  • cyclohexanecarbonyl chloride 136 mg, 0.93 mmol
  • the product was indicated present via UPLC analysis.
  • the mixture was quenched with crushed ice, the extracted with ethyl acetate (2x 10 mL). The organic layer was washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10 ⁇ m column.
  • the mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(benzo[d]thiazol-2-yl)-N-(pyridin-2- ylmethyl)cyclohexanecarboxamide as a white solid (78.4 mg, 0.22 mmol, 36 %).
  • the reaction was warmed to room temperature over 1 hour.
  • the product was indicated present via UPLC analysis.
  • the reaction mixture was extracted with dichloromethane (2x 30 mL), washed with brine (30 mL), dried, concentrated under reduced pressure.
  • the obtained solid was purified by pre-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston pHlex ODS 10um 21.2 ⁇ 250mm 120 ⁇ .
  • the mobile phase was acetonitrile/0.1% aqueous formic acid) to afford the desired compound N-(6-ethylbenzo[d]thiazol-2- yl)-N-(pyridin-4-ylmethyl)cyclohexanecarboxamide (72.9 mg, 0.19 mmol, 38 %) as yellow solid.
  • the elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(5-fluorobenzo[d]thiazol-2-yl)-3-(phenylsulfonyl)-N-(pyridin-3- ylmethyl)propanamide (40.0 mg, 0.090 mmol, 23 %) as a white solid.
  • the obtained solid was purified by pre-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston pHlex ODS 10um 21.2 ⁇ 250mm 120 ⁇ .
  • the mobile phase was acetonitrile/0.1% aqueous formic acid) to afford desire compound N-(6-fluorobenzo[d]thiazol-2-yl)-3-(phenylsulfonyl)-N-(pyridin-3-ylmethyl)propanamide (38.2 mg, 0.080 mmol, 12 %) as a white solid.
  • the mobile phase was acetonitrile/0.1% aqueous formic acid) to afford the desire compound N-(6-ethyl-1,3-benzothiazol-2- yl)-N-[(pyridin-2-yl)methyl]cyclohexanecarboxamide (49.8 mg, 0.13 mmol, 22 %) as a white solid.
  • Example 75 Preparation of 3-(benzenesulfonyl)-N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]propenamide
  • Step 1 Preparation of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde (1.27 g, 12 mmol)
  • 4-fluorobenzo[d]thiazol-2-amine (2.00 g, 12 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of 3-(benzenesulfonyl)-N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]propanamide
  • 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 300 mg, 1.2 mmol
  • 3-(phenylsulfonyl)propanoic acid 372 mg, 1.7 mmol
  • triethylamine 176 mg, 1.7 mmol
  • di(tert-butyl) carbonate 379 mg, 1.7 mmol
  • 4-(dimethylamino)pyridine 212 mg, 1.7 mmol.
  • the mixture was stirred at 50 °C for 17 hours.
  • the product was indicated present via UPLC analysis.
  • the mixture was concentrated under reduced pressure.
  • the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10 ⁇ m column.
  • the mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product 3-(benzenesulfonyl)-N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]propenamide as a white solid (17.6 mg, 0.040 mmol, 3.3 %).
  • the product was indicated present via UPLC analysis.
  • the reaction mixture was filtered.
  • the filtrate was concentrated under reduced pressure and purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston pHlex ODS 10um 21.2 ⁇ 250mm 120 ⁇ .
  • the mobile phase was acetonitrile/0.1% aqueous formic acid) to afford the desire compound 4-cyano-N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-4-ylmethyl)benzamide (45.5 mg, 0.11 mmol, 19 %) as a white solid.
  • Step 1 Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-2- yl)methyl]benzamide
  • Step 1 Preparation of 6-ethyl-N-(pyridin-2-ylmethyl)benzo[d]thiazol-2-amine
  • picolinaldehyde (1.06 g, 9.0 mmol)
  • 6-ethylbenzo[d]thiazol-2-amine (1.60 g, 9.0 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-2-yl)methyl]benzamide
  • 6-ethyl-N-(pyridin-2-ylmethyl)benzo[d]thiazol-2-amine 160 mg, 0.60 mmol
  • 4- cyanobenzoic acid 176 mg, 1.2 mmol
  • N,N-diisopropylethylamine 232 mg, 1.8 mmol
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol) in N,N-dimethylformamide (3 mL) was stirred at room temperature for 2 hours.
  • the product was indicated present via UPLC analysis.
  • the reaction mixture was filtered and concentrated under reduced pressure.
  • Crude product was purified by pre-HPLC (the crude samples were dissolved in N,N-dimethylformamide unless otherwise noted before purification.
  • the mobile phase was acetonitrile/0.1% aqueous formic acid) to afford desire compound 4-cyano-N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-2-ylmethyl)benzamide (75.7 mg, 0.18 mmol, 32 %) as yellow solid.
  • Example 78 Preparation of 4-cyano-N-(5,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
  • Step 1 Preparation of 5,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 343 mg, 3.2 mmol
  • Step 2 Preparation of 4-cyano-N-(5,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • the product was indicated present via UPLC analysis.
  • the mixture was poured into water, then extracted with ethyl acetate (2x 100 mL). The combined organic phase was concentrated under reduced pressure.
  • the crude product was purified by silica gel column chromatography using a gradient of 0-80% ethyl acetate in petroleum ether, then triturated with methanol (10 mL) to afford the target compound 4- cyano-N-(5,6-difluorobenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (0.0324 g, 0.080 mmol, 14 %) as a grey solid.
  • Step 1 Preparation of N-(5,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 5,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 343 mg, 3.2 mmol
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(5,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • a mixture of 5,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 200 mg, 0.22 mmol
  • cyclohexanecarbonyl chloride 256 mg, 1.8 mmol
  • 4-(dimethylamino)pyridine 20.0 mg, 0.16 mmol
  • pyridine 8 mL
  • Step 1 Preparation of N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine
  • 1-trityl-1H-imidazole-4-carbaldehyde (2.50 g, 7.4 mmol) in toluene (20 mL) was added dried 4 ⁇ molecular sieves.
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • a solution of sodium borohydride (1.40 g, 37 mmol) in ethanol (7.4 mL) was added. The reaction was stirred at 110 °C for 1 hour.
  • Step 2 Preparation of N-(benzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide
  • N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine 800 mg, 1.7 mmol
  • N,N-diisopropylethylamine 660 mg, 5.1 mmol
  • dichloromethane 20 mL
  • cyclohexanecarbonyl chloride 296 mg, 2.0 mmol
  • Step 3 Preparation of N-(1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]cyclohexanecarboxamide
  • N-(benzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide 100 mg, 0.17 mmol
  • dichloromethane (1 mL)
  • trifluoroacetic acid (1 mL) at 0 °C.
  • the reaction was warmed to room temperature over 1 hour.
  • the product was indicated present via UPLC analysis.
  • the reaction mixture was concentrated under reduced pressure.
  • the obtained solid was purified by pre-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston pHlex ODS 10um 21.2 ⁇ 250mm 120A.
  • the mobile phase was acetonitrile/0.1% aqueous formic acid) afford desired product N-((1H-imidazol-4-yl)methyl)-N- (benzo[d]thiazol-2-yl)cyclohexanecarboxamide (15.1 mg, 0.040 mmol, 26 %) as a white solid.
  • Example 81 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3- yl)methyl]acetamide
  • To a solution of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (200 mg, 0.77 mmol) in tetrahydrofuran (20 mL) was added sodium hydride (61.6 mg, 1.5 mmol) under argon at 0 °C.
  • the mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(4-fluorobenzo[d]thiazol-2-yl)-2-phenyl-N-(pyridin-3- ylmethyl)acetamide as a white solid (82.1 mg, 0.22 mmol, 28 %).
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (68.0 mg, 1.8 mmol) in dry ethyl alcohol (7.8 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of tert-butyl 2- ⁇ N-[(pyridin-3-yl)methyl]cyclohexaneamido ⁇ -4H,5H,6H,7H- [1,3]thiazolo[5,4-c]pyridine-5-carboxylate
  • tert-butyl 2- ⁇ [(pyridin-3-yl)methyl]amino ⁇ -4H,5H,6H,7H-[1,3]thiazolo[5,4- c]pyridine-5-carboxylate 189 mg, 0.54 mmol
  • dipotassium carbonate (112 mg, 0.81 mmol
  • Step 2 Preparation of N- ⁇ 5-methyl-4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin-2-yl ⁇ -N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • Step 2 Preparation of N- ⁇ 5-ethyl-4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin-2-yl ⁇ -N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • acetaldehyde (0.035 mL, 0.22 mmol), acetic acid (0.013 mL, 0.22 mmol), and sodium triacetoxyborohydride (37.5 mg, 0.18 mmol) were added and the reaction was stirred at room temperature. After 5 minutes, additional sodium triacetoxyborohydride (37.5 mg, 0.18 mmol) was added and the reaction was stirred for 6 hours. The product was indicated present via UPLC analysis.1 M sodium hydroxide and saturated sodium bicarbonate were added to quench the reaction. The reaction was washed with 2x 20 mL portions of dichloromethane and the organic layers were separated and concentrated under reduced pressure.
  • Step 1 Preparation of N-(6-cyano-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 2-(pyridin-3-ylmethylamino)benzo[d]thiazole-6-carbonitrile
  • 2-aminobenzo[d]thiazole-6-carbonitrile 400 mg, 2.3 mmol
  • nicotinaldehyde 244 mg, 2.3 mmol
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(6-cyano-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide
  • 2-(pyridin-3-ylmethylamino)benzo[d]thiazole-6-carbonitrile 100 mg, 0.37 mmol
  • dichloromethane 10 mL
  • N,N-diisopropylethylamine 143 mg, 1.1 mmol
  • the reaction was stirred at 0 °C for 10 min.
  • cyclohexanecarbonyl chloride (80.3 mg, 0.55 mmol) was slowly added.
  • the reaction was stirred at 10 °C for 2 hours.
  • the product was indicated present via UPLC analysis.
  • the mixture was filtered and the solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10 ⁇ m column.
  • the mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(6-cyanobenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide as a white solid (31.4 mg, 0.080 mmol, 23 %).
  • Step 1 Preparation of N-(4-chloro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 4-chlorobenzo[d]thiazol-2-amine
  • 4-chlorobenzo[d]thiazol-2-amine 600 mg, 3.3 mmol
  • nicotinaldehyde 348 mg, 3.3 mmol
  • the mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(4-chloro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide
  • dichloromethane 15 mL
  • N,N-diisopropylethylamine 139 mg, 1.1 mmol
  • cyclohexanecarbonyl chloride 52.6 mg, 0.36 mmol
  • the product was indicated present via UPLC analysis.
  • the mixture was filtered and the solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10 ⁇ m column.
  • the mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(4-chlorobenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide as a white solid (28.4 mg, 0.070 mmol, 21 %).
  • Example 87 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(4-methylpyridin-3- yl)methyl]cyclohexanecarboxamide Step 1: Preparation of 6-ethyl-N-[(4-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of 4-methylpyridine-3-carbaldehyde (149 mg, 1.2 mmol) and 6-ethyl- 1,3-benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 ⁇ molecular sieves under N2 for 24 hours.
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(4-methylpyridin-3- yl)methyl]cyclohexanecarboxamide
  • triethylamine 0.072 mL, 0.52 mmol
  • 6-ethyl-N-[(4- methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine 147 mg, 0.52 mmol
  • dichloromethane (1 mL)
  • cyclohexanecarbonyl chloride (113 mg, 0.78 mmol) was added dropwise, and the reaction was warmed to room temperature and stir for 45 minutes.
  • the product was indicated present via UPLC analysis.
  • the reaction was diluted with 10 mL deionized water and the layers were separated.
  • the inorganic layer was washed with 2x 10 mL washes of dichloromethane.
  • the organic layers were pooled and concentrated under reduced pressure.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes.
  • Step 1 Preparation of N-(6-methanesulfonyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 6-(methylsulfonyl)-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • 6-(methylsulfonyl)benzo[d]thiazol-2-amine 600 mg, 2.6 mmol
  • nicotinaldehyde (281 mg, 2.6 mmol) in toluene (20 mL) was added dried 4 ⁇ molecular sieves.
  • Step 2 Preparation of N-(6-methanesulfonyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • 6-(methylsulfonyl)-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 100 mg, 0.31 mmol
  • dichloromethane 10 mL
  • N,N-diisopropylethylamine 120 mg, 0.93 mmol
  • the mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(6-(methylsulfonyl)benzo[d]thiazol- 2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide as a white solid (3.30 mg, 0.020 mmol, 2.5 %).
  • Example 89 Preparation of ethyl 2- ⁇ N-[(pyridin-3-yl)methyl]cyclohexaneamido ⁇ -1,3-benzothiazole- 6-carboxylate
  • Step 1 Preparation of ethyl 2-(pyridin-3-ylmethylamino)benzo[d]thiazole-6-carboxylate
  • a solution of ethyl 2-aminobenzo[d]thiazole-6-carboxylate (800 mg, 3.6 mmol) and nicotinaldehyde (385 mg, 3.6 mmol) in toluene (16 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of ethyl 2- ⁇ N-[(pyridin-3-yl)methyl]cyclohexaneamido ⁇ -1,3-benzothiazole-6- carboxylate
  • ethyl 2-(pyridin-3-ylmethylamino)benzo[d]thiazole-6-carboxylate 440 mg, 1.4 mmol
  • dichloromethane 15 mL
  • N,N-diisopropylethylamine 542 mg, 4.2 mmol
  • the reaction was stirred at 0 °C for 10 min.
  • cyclohexanecarbonyl chloride (307 mg, 2.1 mmol) was slowly added.
  • the mixture was stirred at 40 °C for 17 hours.
  • the product was indicated present via UPLC analysis.
  • the mixture was filtered and the solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10 ⁇ m column.
  • the mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product ethyl 2-(N-(pyridin-3- ylmethyl)cyclohexanecarboxamido)benzo[d]thiazole-6-carboxylate as a white solid (180 mg, 0.42 mmol, 30 %).
  • Example 90 Preparation of N-[(pyridin-3-yl)methyl]-N-(quinolin-2-yl)cyclohexanecarboxamide
  • 1-(pyridin-3-yl)methanamine 989 mg, 9.2 mmol
  • 2-chloroquinoline 300 mg, 1.8 mmol
  • N,N-diisopropylethylamine 1.59 mL, 9.2 mmol
  • N-Methyl-2-pyrrolidone 9.2 mL
  • Step 2 Preparation of N-[(pyridin-3-yl)methyl]-N-(quinolin-2-yl)cyclohexanecarboxamide
  • N-[(pyridin-3-yl)methyl]quinolin-2-amine 176 mg, 0.74 mmol
  • triethylamine 0.103 mL, 0.74 mmol
  • dichloromethane 3.17 mL
  • cyclohexanecarbonyl chloride 0.147 mL, 1.1 mmol
  • Step 1 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(4-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 6-ethyl-N-[(4-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
  • 4-methoxypyridine-3-carbaldehyde 168 mg, 1.2 mmol
  • 6- ethyl-1,3-benzothiazol-2-amine 200 mg, 1.1 mmol
  • dry toluene 11 mL
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(4-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide
  • triethylamine 8. mg, 0.81 mmol
  • 6-ethyl-N-[(4- methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (244 mg, 0.81 mmol) in dichloromethane (1 mL) and stirred at 0 °C.
  • cyclohexanecarbonyl chloride (177 mg, 1.2 mmol) was added dropwise, and the reaction was warmed to room temperature and stir for 45 minutes.
  • the product was indicated present via UPLC analysis.
  • the reaction was diluted with 10 mL deionized water and the layers were separated.
  • the inorganic layer was washed with 2x 10 mL washes of dichloromethane.
  • the organic layers were pooled and concentrated under reduced pressure.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes.
  • Example 92 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(5-fluoropyridin-3- yl)methyl]cyclohexanecarboxamide Step 1: Preparation of 6-ethyl-N-[(5-fluoropyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of 5-fluoropyridine-3-carbaldehyde 153 mg, 1.2 mmol
  • 6-ethyl- 1,3-benzothiazol-2-amine 200 mg, 1.1 mmol
  • dry toluene 11 mL
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(5-fluoropyridin-3- yl)methyl]cyclohexanecarboxamide
  • triethylamine (0.18 mL, 1.3 mmol) was added to a solution of 6-ethyl-N-[(5- fluoropyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (321 mg, 1.1 mmol) in dichloromethane (1.1 mL) and stirred at 0 °C.
  • cyclohexanecarbonyl chloride (243 mg, 1.7 mmol) was added dropwise, and the reaction was warmed to room temperature and stir for 45 minutes.
  • the product was indicated present via UPLC.
  • the reaction was diluted with 10 mL deionized water and the layers were separated.
  • the inorganic layer was washed with 2x 10 mL washes of dichloromethane.
  • the organic layers were pooled and concentrated under reduced pressure.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-50% ethyl acetate in hexanes.
  • Step 1 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-5- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 6-ethyl-N-[(1H-pyrazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of 1H-pyrazole-5-carbaldehyde 118 mg, 1.2 mmol
  • 6-ethyl-1,3- benzothiazol-2-amine 200 mg, 1.1 mmol
  • dry toluene 11 mL
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-5- yl)methyl]cyclohexanecarboxamide
  • triethylamine (0.16 mL, 1.1 mmol) was added to a solution of 6-ethyl-N-[(1H- pyrazol-5-yl)methyl]-1,3-benzothiazol-2-amine (248 mg, 0.96 mmol) in dichloromethane (1 mL) and stirred at 0 °C.
  • cyclohexanecarbonyl chloride (209 mg, 1.4 mmol) was added dropwise, and the reaction was warmed to room temperature and stir for 2 hours.
  • the product was confirmed present via UPLC analysis.
  • the reaction was diluted with 10 mL deionized water and the layers were separated.
  • the inorganic layer was washed with 2x 10 mL portions of dichloromethane.
  • the organic layers were pooled and concentrated under reduced pressure.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-50% ethyl acetate in hexanes.
  • Example 94 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]bicyclo[1.1.1]pentane-1-carboxamide
  • a solution of bicyclo[1.1.1]pentane-1-carboxylic acid (67.0 mg, 0.60 mmol) in thionyl chloride (2 mL) was stirred at 70 °C for 2 hours.
  • the reaction was concentrated under reduced pressure and re- dissolved in dichloromethane (2 mL).6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (80.0 mg, 0.30 mmol) and N,N-diisopropylethylamine (77.0 mg, 0.60 mmol) was added to the solution at 0 °C. The reaction was warmed to room temperature with stirring over 16 hours. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane (20 mL) and washed with water (20 mL). The organic layer separated and concentrated under reduced pressure.
  • Crude product was purified by Prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10 ⁇ m column. The mobile phase was acetonitrile/0.01% aqueous formic acid) obtaining N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)bicyclo[1.1.1]pentane-1-carboxamide (24.0 mg, 0.070 mmol, 22 %) as white solid.
  • the mobile phase was acetonitrile/0.01% aqueous formic acid) obtaining N-(6-ethylbenzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)tetrahydro-2H-pyran-4-carboxamide (62.0 mg, 0.16 mmol, 54 %) as white solid.
  • Example 96 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-4-phenyl-N-[(pyridin-3- yl)methyl]butanamide
  • Step 1 Preparation of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 481 mg, 4.5 mmol
  • 5-fluorobenzo[d]thiazol-2-amine 500 mg, 3.0 mmol
  • toluene (20 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-4-phenyl-N-[(pyridin-3-yl)methyl]butanamide
  • 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 100 mg, 0.39 mmol
  • N,N-dimethylformamide 5 mL
  • 4-phenylbutanoic acid 127 mg, 0.77 mmol
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (292 mg, 0.77 mmol)
  • N,N-diisopropylethylamine 115 mg, 0.77 mmol.
  • Step 1 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-5- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 6-fluoro-N-[(1H-pyrazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of 1H-pyrazole-5-carbaldehyde 123 mg, 1.3 mmol
  • 6-fluoro-1,3- benzothiazol-2-amine 200 mg, 1.2 mmol
  • dry toluene (12 mL) was refluxed at 120 °C with activated 4 ⁇ molecular sieves under N2 for 24 hours.
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry ethyl alcohol (12 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-5- yl)methyl]cyclohexanecarboxamide
  • triethylamine 99.6 mg, 0.99 mmol
  • 6-fluoro-N-[(1H- pyrazol-5-yl)methyl]-1,3-benzothiazol-2-amine 204 mg, 0.82 mmol
  • cyclohexanecarbonyl chloride 180 mg, 1.2 mmol was added dropwise, and the reaction was warmed to room temperature and stir for 1.5 hours.
  • the product was indicated present via UPLC analysis.
  • the reaction was diluted with 10 mL deionized water and the layers were separated.
  • the inorganic layer was washed with 2x 10 mL washes of dichloromethane.
  • the organic layers were pooled and concentrated under reduced pressure.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-50% ethyl acetate in hexanes.
  • Example 98 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-4-methyl-N-[(pyridin-3- yl)methyl]piperazine-1-carboxamide
  • Step 1 Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 2.70 g, 25 mmol
  • 6-ethylbenzo[d]thiazol-2-amine 3.00 g, 17 mmol
  • toluene 60 mL
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-4-methyl-N-[(pyridin-3-yl)methyl]piperazine-1- carboxamide
  • triphosgene 65.3 mg, 0.22 mmol
  • dichloromethane 10 mL
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 120 mg, 0.45 mmol
  • pyridine 142 mg, 1.8 mmol
  • the mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(6- ethylbenzo[d]thiazol-2-yl)-4-methyl-N-(pyridin-3-ylmethyl)piperazine-1-carboxamide as a white solid (17.6 mg, 0.040 mmol, 9.9 %).
  • Example 99 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclopentanecarboxamide
  • Step 1 Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 2.70 g, 25 mmol
  • 6-ethylbenzo[d]thiazol-2-amine 3.00 g, 17 mmol
  • toluene 60 mL
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclopentanecarboxamide
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 120 mg, 0.45 mmol
  • dichloromethane 10 mL
  • N,N-diisopropylethylamine 174 mg, 1.4 mmol
  • the solution was stirred at 0 °C for 10 min, then cyclopentanecarbonyl chloride (88.8 mg, 0.67 mmol) was slowly added.
  • Example 100 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclobutanecarboxamide
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 80.0 mg, 0.30 mmol
  • N,N-diisopropylethylamine 80.0 mg, 0.60 mmol
  • dichloromethane 4 mL
  • the reaction was warmed to room temperature with stirring over 16 hours.
  • the product was indicated present via UPLC analysis. Water (30 mL) was added, then the mixture was extracted with dichloromethane (2x 30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure.
  • the crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10 ⁇ m column.
  • the mobile phase was acetonitrile/0.01% aqueous formic acid) to give N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclobutanecarboxamide (40.0 mg, 0.11 mmol, 38 %) as a white solid.
  • Example 101 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-2-methyl-N-[(pyridin-3- yl)methyl]propenamide
  • Isobutyryl chloride (64.0 mg, 0.60 mmol) was added into a solution of 6-ethyl-N-(pyridin-3- ylmethyl)benzo[d]thiazol-2-amine (80.0 mg, 0.30 mmol) and N,N-diisopropylethylamine (77.0 mg, 0.60 mmol) in dichloromethane (2 mL) at 0 °C.
  • Example 102 Preparation of N-(5,6-dimethoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide Step 1: Preparation of 5,6-dimethoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine To a solution of nicotinaldehyde (0.890 g, 8.4 mmol), 5,6-dimethoxybenzo[d]thiazol-2-amine (1.20 g, 5.6 mmol) in toluene (20 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(5,6-dimethoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • a solution of 5,6-dimethoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 300 mg, 1.0 mmol
  • cyclohexanecarboxylic acid (128 mg, 1.0 mmol)
  • triethylamine 303 mg, 3.0 mmol
  • dichloromethane 10 mL
  • N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide 570 mg, 1.5 mmol).
  • Example 103 Preparation of 2-(benzyloxy)-N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]acetamide
  • Step 1 Preparation of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 481 mg, 4.5 mmol
  • 5-fluorobenzo[d]thiazol-2-amine 500 mg, 3.0 mmol
  • toluene (20 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of 2-(benzyloxy)-N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]acetamide
  • 2-(benzyloxy)acetic acid 256 mg, 1.5 mmol
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (585 mg, 1.5 mmol) and N,N-diisopropylethylamine (198 mg, 1.5 mmol).
  • Step 1 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-1,1-dioxo-N-[(pyridin-3-yl)methyl]- 1 ⁇ 6-thiomorpholine-4-carboxamide
  • Step 1 Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 2.70 g, 25 mmol
  • 6-ethylbenzo[d]thiazol-2-amine 3.00 g, 17 mmol
  • toluene 60 mL
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-1,1-dioxo-N-[(pyridin-3-yl)methyl]-1 ⁇ 6- thiomorpholine-4-carboxamide
  • triphosgene 65.3 mg, 0.22 mmol
  • dichloromethane 10 mL
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 120 mg, 0.45 mmol
  • pyridine 142 mg, 1.8 mmol
  • the mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(6- ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)thiomorpholine-4-carboxamide 1,1-dioxide (43.6 mg, 0.10 mmol, 23 %) as a white solid.
  • Step 1 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]oxane-3- carboxamide
  • Step 1 Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 2.70 g, 25 mmol
  • 6-ethylbenzo[d]thiazol-2-amine 3.00 g, 17 mmol
  • toluene 60 mL
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]oxane-3-carboxamide
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 120 mg, 0.45 mmol
  • dichloromethane 15 mL
  • tetrahydro-2H-pyran-3-carboxylic acid 87.1 mg, 0.67 mmol
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide 255 mg, 0.67 mmol
  • triethylamine 136 mg, 1.4 mmol
  • Example 106 Preparation of 4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]benzamide
  • Step 1 Preparation of 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • 6-fluorobenzo[d]thiazol-2-amine 3.00 g, 18 mmol
  • toluene 50 mL
  • Step 2 Preparation of 4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]benzamide
  • N-((1H-imidazol-4-yl)methyl)-6-fluorobenzo[d]thiazol-2-amine 200 mg, 0.81 mmol
  • 4-cyanobenzoic acid 236 mg, 1.6 mmol
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (611 mg, 1.6 mmol) and N,N-diisopropylethylamine (207 mg, 1.6 mmol).
  • Step 2 Preparation of tert-butyl 4-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3- yl)methyl]carbamoyl]piperidine-1-carboxylate
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 300 mg, 1.1 mmol
  • dichloromethane 15 mL
  • 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (380 mg, 1.7 mmol)
  • N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide 631 mg, 1.7 mmol
  • triethylamine 336 mg, 3.3 mmol
  • Example 108 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-4- yl)methyl]cyclohexanecarboxamide Step 1: Preparation of 6-fluoro-N-[(1H-pyrazol-4-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of 1H-pyrazole-4-carbaldehyde 113 mg, 1.2 mmol
  • 6-fluoro-1,3- benzothiazol-2-amine 200 mg, 1.2 mmol
  • dry toluene (12 mL) was refluxed at 120 °C with activated 4 ⁇ molecular sieves under N2 for 24 hours.
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry ethyl alcohol (12 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-4- yl)methyl]cyclohexanecarboxamide
  • 6-fluoro-N-[(1H-pyrazol-4-yl)methyl]-1,3-benzothiazol-2-amine 211 mg, 0.85 mmol
  • triethylamine 0.17 mL, 1.3 mmol
  • Example 109 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-4- yl)methyl]cyclohexanecarboxamide Step 1: Preparation of 6-ethyl-N-[(1H-pyrazol-4-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of 1H-pyrazole-4-carbaldehyde 107 mg, 1.1 mmol
  • 6-ethyl-1,3- benzothiazol-2-amine 200 mg, 1.1 mmol
  • dry toluene 11 mL
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-4- yl)methyl]cyclohexanecarboxamide
  • 6-ethyl-N-[(1H-pyrazol-4-yl)methyl]-1,3-benzothiazol-2-amine (217 mg, 0.84 mmol) and triethylamine (0.17 mL, 1.3 mmol) were dissolved in tetrahydrofuran (1 mL) and cooled to 0 °C.
  • cyclohexanecarbonyl chloride (244 mg, 1.7 mmol) was added dropwise with stirring.
  • Example 110 Preparation of N-[(pyridin-3-yl)methyl]-N-(4,5,6,7-tetrahydro-1,3-benzothiazol-2- yl)cyclohexanecarboxamide Step 1: Preparation of 4,5,6,7-tetrahydro-1,3-benzothiazol-2-amine
  • a solution of pyridine-3-carbaldehyde (138 mg, 1.3 mmol) and 4,5,6,7- tetrahydro-1,3-benzothiazol-2-amine 200 mg, 1.3 mmol
  • dry toluene 13 mL
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (111 mg, 3.0 mmol) in dry ethyl alcohol (13 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of N-[(pyridin-3-yl)methyl]-N-(4,5,6,7-tetrahydro-1,3-benzothiazol-2- yl)cyclohexanecarboxamide
  • N-[(pyridin-3-yl)methyl]-4,5,6,7-tetrahydro-1,3-benzothiazol-2-amine 256 mg, 1.0 mmol
  • triethylamine (0.21 mL, 1.5 mmol) were dissolved in tetrahydrofuran (1.03 mL) and cooled to 0 °C.
  • cyclohexanecarbonyl chloride (302 mg, 2.1 mmol) was added dropwise with stirring.
  • Step 1 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperidine-1- carboxamide
  • Step 1 Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 2.70 g, 25 mmol
  • 6-ethylbenzo[d]thiazol-2-amine 3.00 g, 17 mmol
  • toluene 60 mL
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperidine-1-carboxamide
  • triphosgene 65.3 mg, 0.22 mmol
  • dichloromethane 10 mL
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 120 mg, 0.45 mmol
  • pyridine 142 mg, 1.8 mmol
  • Example 112 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]morpholine-4- carboxamide Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]morpholine-4-carboxamide
  • triphosgene 65.3 mg, 0.20 mmol
  • dichloromethane 10 mL
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 120 mg, 0.45 mmol
  • pyridine 142 mg, 1.8 mmol
  • Example 113 Preparation of N-(6-ethoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide Step 1: Preparation of 6-ethoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine To a solution of nicotinaldehyde (1.07 g, 10 mmol), 6-ethoxybenzo[d]thiazol-2-amine (1.94 g, 10 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(6-ethoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • 6-ethoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 200 mg, 0.70 mmol
  • N,N-diisopropylethylamine 180 mg, 1.4 mmol
  • dichloromethane 5 mL
  • cyclohexanecarbonyl chloride 210 mg, 1.4 mmol
  • Step 1 Preparation of N-(6-bromo-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 6-bromo-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 1.07 g, 10 mmol
  • 6-bromobenzo[d]thiazol-2-amine (2.29 g, 10 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(6-bromo-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide
  • 6-bromo-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 200 mg, 0.60 mmol
  • N,N-diisopropylethylamine 160 mg, 1.3 mmol
  • dichloromethane 5 mL
  • cyclohexanecarbonyl chloride 180 mg, 1.3 mmol
  • Example 115 Preparation of N-(7-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3- yl)methyl]propenamide
  • Step 1 Preparation of 7-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 321 mg, 3.0 mmol
  • 7-fluorobenzo[d]thiazol-2-amine 3m40 g, 2.0 mmol
  • Step 2 Preparation of N-(7-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3-yl)methyl]propanamide
  • N,N-dimethylformamide 5 mL
  • 3-phenoxypropanoic acid 166 mg, 1.0 mmol
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (285 mg, 0.75 mmol) and triethylamine (150 mg, 1.5 mmol).
  • Step 1 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1-methyl-1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 6-fluoro-N-((1-methyl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine
  • 6- fluorobenzo[d]thiazol-2-amine (1.00 g, 6.0 mmol) in toluene (30 mL) was added dried 4 ⁇ molecular sieves.
  • Step 2 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1-methyl-1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
  • 6-fluoro-N-((1-methyl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine 100 mg, 0.38 mmol
  • N,N-dimethylformamide 10 mL
  • cyclohexanecarboxylic acid 73.0 mg, 0.57 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide 216 mg, 1.6 mmol
  • Step 2 Preparation of tert-butyl 6-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3-yl)methyl]carbamoyl]-2- azaspiro[3.3]heptane-2-carboxylate
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (0.600 g, 2.2 mmol) in dichloromethane (20 mL) were added 2-(tert-butoxycarbonyl)-2-azaspiro[3.3]heptane-6-carboxylic acid (0.805 g, 3.3 mmol)
  • N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (1.30 g, 3.3 mmol) and trieth
  • Example 119 Preparation of N-(1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of 1H-imidazole-5-carbaldehyde (140 mg, 1.5 mmol) and 1,3- benzothiazol-2-amine (200 mg, 1.3 mmol) in dry toluene (13 mL) was refluxed at 120 °C with activated 4 ⁇ molecular sieves under N2 for 24 hours.
  • Step 2 Preparation of N-(1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]cyclohexanecarboxamide
  • N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine (306 mg, 1.3 mmol) and triethylamine (0.46 mL, 3.3 mmol) were dissolved in tetrahydrofuran (1.32 mL) and cooled to 0 °C.
  • cyclohexanecarbonyl chloride (580 mg, 4.0 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight.
  • Step 1 Preparation of N-[(pyridin-3-yl)methyl]-N-(quinazolin-2-yl)cyclohexanecarboxamide
  • Step 1 Preparation of N-[(pyridin-3-yl)methyl]quinazolin-2-amine
  • a solution of pyridine-3-carbaldehyde (0.14 mL, 1.5 mmol) and quinazolin-2- amine (200 mg, 1.4 mmol) in dry toluene (14 mL) was stirred at 100 °C with activated 4 ⁇ molecular sieves under nitrogen overnight.
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (119 mg, 3.2 mmol) in dry ethyl alcohol (14 mL). The reaction mixture was stirred at 70 °C for 30 min and then cooled to room temperature. The product was confirmed vial UPLC analysis. The reaction was filtered over a bed of Celite and washed with ethyl acetate (2x 20 mL). Crude product was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane through 24 g of silica gel.
  • Step 2 Preparation of N-[(pyridin-3-yl)methyl]-N-(quinazolin-2-yl)cyclohexanecarboxamide
  • N-[(pyridin-3-yl)methyl]quinazolin-2-amine 76.9 mg, 0.33 mmol
  • tetrahydrofuran 1.6 mL
  • cyclohexanecarbonyl chloride 0.065 mL, 0.49 mmol
  • Step 1 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 6-ethyl-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of pyrimidine-5-carbaldehyde (121 mg, 1.1 mmol) and 6-ethyl-1,3- benzothiazol-2-amine 200 mg, 1.1 mmol
  • dry toluene 11 mL
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]cyclohexanecarboxamide
  • 6-ethyl-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine (45.6 mg, 0.17 mmol) and triethylamine (0.035 mL, 0.25 mmol) were dissolved in tetrahydrofuran (1 mL) and cooled to 0 °C.
  • cyclohexanecarbonyl chloride 36.9 mg, 0.25 mmol
  • Step 1 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 6-ethyl-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of 1,3-thiazole-5-carbaldehyde 140 mg, 1.2 mmol
  • 6-ethyl-1,3- benzothiazol-2-amine (219 mg, 1.2 mmol) in dry toluene (12 mL) was refluxed at 120 °C with activated 4 ⁇ molecular sieves under N2 for 24 hours.
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (106 mg, 2.8 mmol) in dry ethyl alcohol (12 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5- yl)methyl]cyclohexanecarboxamide
  • 6-ethyl-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine (241 mg, 0.87 mmol) and triethylamine (0.18 mL, 1.3 mmol) were dissolved in tetrahydrofuran (1.7 mL) and cooled to 0 °C.
  • cyclohexanecarbonyl chloride 190 mg, 1.3 mmol
  • Step 1 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 6-fluoro-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • 1,3-thiazole-5-carbaldehyde 242 mg, 2.1 mmol
  • 6-fluoro-1,3- benzothiazol-2-amine 360 mg, 2.1 mmol
  • dry toluene 21 mL
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (186 mg, 4.9 mmol) in dry ethyl alcohol (21 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5- yl)methyl]cyclohexanecarboxamide
  • 6-fluoro-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine (393 mg, 1.5 mmol) and triethylamine (0.31 mL, 2.2 mmol) were dissolved in tetrahydrofuran (3 mL) and cooled to 0 °C.
  • cyclohexanecarbonyl chloride (324 mg, 2.2 mmol) was added dropwise with stirring.
  • Example 125 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]oxolane-2- carboxamide
  • Step 1 Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 2.70 g, 25 mmol
  • 6-ethylbenzo[d]thiazol-2-amine 3.00 g, 17 mmol
  • toluene 60 mL
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]oxolane-2-carboxamide
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 120 mg, 0.45 mmol
  • dichloromethane 15 mL
  • tetrahydrofuran-2-carboxylic acid 77.7 mg, 0.67 mmol
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide 255 mg, 0.67 mmol
  • triethylamine 136 mg, 1.4 mmol
  • Example 126 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]pyrrolidine-1- carboxamide
  • Step 1 Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 2.70 g, 25 mmol
  • 6-ethylbenzo[d]thiazol-2-amine 3.00 g, 17 mmol
  • toluene 60 mL
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]pyrrolidine-1-carboxamide
  • triphosgene 65.3 mg, 0.22 mmol
  • dichloromethane 10 mL
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 120 mg, 0.45 mmol
  • pyridine 142 mg, 1.8 mmol
  • Example 127 Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 6-chloro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 0.90 g, 8.4 mmol
  • 6-ethylbenzo[d]thiazol-2-amine (1.00 g, 5.6 mmol) in toluene (20 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide
  • 6-chloro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 110 mg, 0.39 mmol
  • triethylamine 78.0 mg, 0.78 mmol
  • dichloromethane 5 mL
  • cyclohexanecarbonyl chloride 85.0 mg, 0.59 mmol
  • Example 128 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(phenylamino)-N-[(pyridin-3- yl)methyl]propenamide
  • Step 1 Preparation of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 481 mg, 4.5 mmol
  • 5-fluorobenzo[d]thiazol-2-amine 500 mg, 3.0 mmol
  • toluene (20 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(phenylamino)-N-[(pyridin-3- yl)methyl]propanamide
  • 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 300 mg, 1.2 mmol
  • N,N-dimethylformamide 15 mL
  • 3-(phenylamino)propanoic acid (286 mg, 1.7 mmol)
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (661 mg, 1.7 mmol) and N,N-diisopropylethylamine (223 mg, 1.7 mmol).
  • Example 129 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5- yl)methyl]cyclohexanecarboxamide Step 1: Preparation of 6-ethyl-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of 1H-imidazole-4-carbaldehyde 107 mg, 1.1 mmol
  • 6-ethyl-1,3- benzothiazol-2-amine 200 mg, 1.1 mmol
  • dry toluene 11 mL
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5- yl)methyl]cyclohexanecarboxamide
  • 6-ethyl-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine (289 mg, 1.1 mmol) and triethylamine (0.23 mL, 1.7 mmol) were dissolved in tetrahydrofuran (2.2 mL) and cooled to 0 °C.
  • cyclohexanecarbonyl chloride 243 mg, 1.7 mmol
  • Example 130 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridazin-3- yl)methyl]cyclohexanecarboxamide Step 1: Preparation of 6-ethyl-N-[(pyridazin-3-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of pyridazine-3-carbaldehyde 121 mg, 1.1 mmol
  • 6-ethyl-1,3- benzothiazol-2-amine 200 mg, 1.1 mmol
  • dry toluene 11 mL
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • cyclohexanecarbonyl chloride (73.8 mg, 0.50 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of dichloromethane were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Example 131 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(2-fluorophenoxy)-N-[(pyridin-3- yl)methyl]propenamide
  • Step 1 Preparation of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 481 mg, 4.5 mmol
  • 5-fluorobenzo[d]thiazol-2-amine 500 mg, 3.0 mmol
  • toluene (20 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(2-fluorophenoxy)-N-[(pyridin-3- yl)methyl]propanamide
  • 1-hydroxybenzotriazole 86.4 mg, 0.64 mmol
  • 1,3-dicyclohexylcarbodiimide 132 mg, 0.64 mmol
  • Step 1 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(3-fluorophenoxy)-N-[(pyridin-3- yl)methyl]propenamide
  • Step 1 Preparation of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 481 mg, 4.5 mmol
  • 5-fluorobenzo[d]thiazol-2-amine 500 mg, 3.0 mmol
  • toluene 20 mL
  • Step 2 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(3-fluorophenoxy)-N-[(pyridin-3- yl)methyl]propanamide
  • 1-hydroxybenzotriazole 86.4 mg, 0.64 mmol
  • 1,3-dicyclohexylcarbodiimide 132 mg, 0.64 mmol
  • Example 133 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(4-fluorophenoxy)-N-[(pyridin-3- yl)methyl]propenamide
  • Step 1 Preparation of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 481 mg, 4.5 mmol
  • 5-fluorobenzo[d]thiazol-2-amine 500 mg, 3.0 mmol
  • toluene (20 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(4-fluorophenoxy)-N-[(pyridin-3- yl)methyl]propanamide
  • 1-hydroxybenzotriazole 86.4 mg, 0.64 mmol
  • 1,3-dicyclohexylcarbodiimide 132 mg, 0.64 mmol
  • Example 134 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]benzamide
  • Step 1 Preparation of 6-ethyl-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • 6-ethylbenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves.
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]benzamide
  • N-((1H-imidazol-4-yl)methyl)-6-ethylbenzo[d]thiazol-2-amine 200 mg, 0.76 mmol
  • benzoic acid 186 mg, 1.5 mmol
  • N-[(dimethylamino)-1H- 1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide 577 mg, 1.5 mmol
  • N,N-diisopropylethylamine (194 mg, 1.5 mmol).
  • Step 1 Preparation of 4-cyano-N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol- 2-yl)benzamide
  • Step 1 Preparation of 6-methoxy-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • 6-methoxybenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves.
  • Step 2 Preparation of 4-cyano-N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2- yl)benzamide
  • N-((1H-imidazol-4-yl)methyl)-6-methoxybenzo[d]thiazol-2-amine 200 mg, 0.76 mmol
  • 4-cyanobenzoic acid 223 mg, 1.5 mmol
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (577 mg, 1.5 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol).
  • Step 1 Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-4-cyano-N-[(1H-imidazol-4- yl)methyl]benzamide
  • Step 1 Preparation of 6-chloro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • 6-chlorobenzo[d]thiazol-2- amine (1.00 g, 5.4 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves.
  • Step 2 Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-4-cyano-N-[(1H-imidazol-4-yl)methyl]benzamide
  • 6-chloro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine 200 mg, 0.75 mmol
  • N,N-dimethylformamide 5 mL
  • 4-cyanobenzoic acid (223 mg, 1.5 mmol)
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (577 mg, 1.5 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol).
  • Step 1 Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3- yl)methyl]acetamide
  • Step 1 Preparation of 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 1 g, 11 mmol
  • 6-methoxybenzo[d]thiazol-2-amine (2.00 g, 11 mmol) in toluene (10 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3-yl)methyl]acetamide
  • 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 200 mg, 0.74 mmol
  • 2-phenylacetic acid 120 mg, 0.89 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (420 mg, 1.1 mmol) and N,N- diisopropylethylamine (286 mg, 2.2 mmol) were dissolved in dichloromethane (5 mL).
  • Step 1 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-2- phenylacetamide
  • Step 1 Preparation of 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • 6-fluorobenzo[d]thiazol-2-amine 3.00 g, 18 mmol
  • toluene 50 mL
  • Step 2 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-2-phenylacetamide
  • 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine 200 mg, 0.81 mmol
  • 2-phenylacetic acid 132 mg, 0.97 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin- 1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (459 mg, 1.2 mmol) and N,N- diisopropylethylamine (312 mg, 2.4 mmol) were dissolved in dichloromethane (5 mL).
  • Step 1 Preparation of N-[(1H-imidazol-5-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2-yl)-2- phenylacetamide
  • Step 1 Preparation of 6-methoxy-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • 6-methoxybenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves.
  • Step 2 Preparation of N-[(1H-imidazol-5-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2-yl)-2- phenylacetamide
  • N-((1H-imidazol-5-yl)methyl)-6-methoxybenzo[d]thiazol-2-amine 230 mg, 0.88 mmol
  • 2-phenylacetic acid 144 mg, 1.1 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (504 mg, 1.3 mmol) and N,N- diisopropylethylamine (342 mg, 2.7 mmol) were dissolved in dichloromethane (5 mL
  • Step 1 Preparation of 4-chloro-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
  • Step 1 Preparation of 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of pyridine-3-carbaldehyde (0.63 mL, 6.7 mmol) and 6-ethyl-1,3-benzothiazol-2-amine (1.20 g, 6.7 mmol) in dry toluene (34 mL) was stirred at 105°C with activated 4 ⁇ molecular sieves under nitrogen overnight.
  • the reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (1.27 g, 34 mmol) in dry ethyl alcohol (34 mL). The reaction mixture was stirred at 70 °C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with ethyl acetate. The filtrate was concentrated then diluted with dichloromethane and filtered again. The filtrate was purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane through a 40 g column of silica gel.
  • Step 2 Preparation of 4-chlorobenzoyl chloride
  • 4-chlorobenzoic acid 300 mg, 1.9 mmol
  • dichloromethane 9.5 mL
  • oxalyl chloride 0.81 mL, 9.5 mmol
  • N,N-dimethylformamide were added as a catalyst and the reaction bubbled.
  • the reaction was then heated to 40 °C and stirred overnight.
  • the product was indicated present via crude NMR analysis.
  • the reaction was cooled to room temperature and concentrated under reduced pressure. Crude product was indicated pure enough to continue to next step via NMR and assumed 100% yield.
  • Step 3 Preparation of 4-chloro-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
  • 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine 400 mg, 1.5 mmol
  • tetrahydrofuran 7.4 mL
  • 4-chlorobenzoyl chloride 334 mg, 1.9 mmol
  • the reaction was stirred from 0 °C to room temperature for 1.5 hours.
  • the product was indicated present via UPLC analysis.
  • the reaction was diluted with dichloromethane and washed with water (3x).
  • the organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated.
  • the residue was dissolved with ethyl acetate and filtered again.
  • Crude product was purified via flash column chromatography eluting with 0-100% hexanes/ethyl acetate through 40 g of silica gel.
  • Step 1 Preparation of N-[(pyridin-3-yl)methyl]-N-(quinoxalin-2-yl)cyclohexanecarboxamide Step 1: Preparation of N-[(pyridin-3-yl)methyl]quinoxalin-2-amine
  • a solution of pyridine-3-carbaldehyde (0.097 mL, 1.0 mmol) and quinoxalin-2- amine (150 mg, 1.0 mmol) in dry toluene (5.2 mL) was stirred at 100 °C with activated 4 ⁇ molecular sieves under nitrogen overnight.
  • the reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (194 mg, 5.2 mmol) in dry ethyl alcohol (5.2 mL). The reaction mixture was stirred at 70 °C for 30 minutes and then cooled to room temperature. The reaction was filtered over a bed of Celite and washed with ethyl acetate (2x 20 mL). The filtrate was concentrated then diluted with dichloromethane and filtered again. The filtrate was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane.
  • Step 2 Preparation of N-[(pyridin-3-yl)methyl]-N-(quinoxalin-2-yl)cyclohexanecarboxamide
  • N-[(pyridin-3-yl)methyl]quinoxalin-2-amine 107 mg, 0.45 mmol
  • tetrahydrofuran 2.2 mL
  • cyclohexanecarbonyl chloride 98.5 mg, 0.67 mmol
  • Step 1 Preparation of (1s,3r)-N-(6-ethyl-1,3-benzothiazol-2-yl)-3-methyl-N-[(pyridin-3- yl)methyl]cyclobutane-1-carboxamide
  • Step 1 Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 2.70 g, 25 mmol
  • 6-ethylbenzo[d]thiazol-2-amine 3.00 g, 17 mmol
  • toluene 60 mL
  • Step 2 Preparation of (1s,3r)-N-(6-ethyl-1,3-benzothiazol-2-yl)-3-methyl-N-[(pyridin-3- yl)methyl]cyclobutane-1-carboxamide
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 120 mg, 0.45 mmol
  • dichloromethane 15 mL
  • 3-methylcyclobutanecarboxylic acid 76.4 mg, 0.67 mmol
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide 255 mg, 0.67 mmol
  • triethylamine 136 mg, 1.4 mmol
  • the reaction was stirred at 10 °C for 17 hours.
  • the product was indicated present via UPLC analysis.
  • the solvent was removed under the reduced pressure and the residue was further purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10 ⁇ m column.
  • the mobile phase was acetonitrile/0.01% aqueous formic acid).
  • Racemic product N-(6- ethylbenzo[d]thiazol-2-yl)-3-methyl-N-(pyridin-3-ylmethyl)cyclobutanecarboxamide (80.0 mg, 0.22 mmol, 49 %) was afforded as a white solid.80 mg of this product was sent to chiral separation (the crude sample was dissolved in 35 mL methanol. AD 20*250 mm column. The mobile phase was 0.2% methanol/ammonia) to afford two isomers.
  • Example 143 Preparation of N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2- yl)cyclohexanecarboxamide Step 1: Preparation of 6-methoxy-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine To a solution of 1H-imidazole-4-carbaldehyde (0.799 g, 8.3 mmol), 6-methoxybenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves.
  • Step 2 Preparation of N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2- yl)cyclohexanecarboxamide
  • N-((1H-imidazol-5-yl)methyl)-6-methoxybenzo[d]thiazol-2-amine 230 mg, 0.88 mmol
  • cyclohexanecarboxylic acid (135 mg, 1.1 mmol)
  • N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5- b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (504 mg, 1.3 mmol) and N,N-diisopropylethylamine (342 mg, 2.6 mmol) were dissolved in
  • Step 1 Preparation of N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2- yl)benzamide
  • Step 1 Preparation of 6-methoxy-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • 6-methoxybenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2-yl)benzamide
  • N-((1H-imidazol-4-yl)methyl)-6-methoxybenzo[d]thiazol-2-amine 200 mg, 0.76 mmol
  • benzoic acid 185 mg, 1.5 mmol
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (577 mg, 1.5 mmol)
  • N,N-diisopropylethylamine (194 mg, 1.5 mmol).
  • Step 1 Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]benzamide
  • Step 1 Preparation of 6-ethyl-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • 6-ethylbenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves.
  • Step 2 Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]benzamide
  • N-((1H-imidazol-4-yl)methyl)-6-ethylbenzo[d]thiazol-2-amine 200 mg, 0.76 mmol
  • 4-cyanobenzoic acid 223 mg, 1.5 mmol
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (577 mg, 1.5 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol).
  • Step 1 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3- yl)methyl]acetamide
  • Step 1 Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 2.70 g, 25 mmol
  • 6-ethylbenzo[d]thiazol-2-amine 3.00 g, 17 mmol
  • toluene 60 mL
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3-yl)methyl]acetamide
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 260 mg, 0.97 mmol
  • 2-phenylacetic acid 158 mg, 1.2 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide 550 mg, 1.5 mmol
  • N,N- diisopropylethylamine 374 mg, 2.9 mmol
  • Step 1 Preparation of (1r,3s)-N-(6-ethyl-1,3-benzothiazol-2-yl)-3-methyl-N-[(pyridin-3- yl)methyl]cyclobutane-1-carboxamide
  • Step 1 Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 2.70 g, 25 mmol
  • 6-ethylbenzo[d]thiazol-2-amine 3.00 g, 17 mmol
  • toluene 60 mL
  • Step 2 Preparation of (1r,3s)-N-(6-ethyl-1,3-benzothiazol-2-yl)-3-methyl-N-[(pyridin-3- yl)methyl]cyclobutane-1-carboxamide
  • 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine 120 mg, 0.45 mmol
  • dichloromethane 15 mL
  • 3-methylcyclobutanecarboxylic acid 76.4 mg, 0.67 mmol
  • N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide 255 mg, 0.67 mmol
  • triethylamine 136 mg, 1.4 mmol
  • the reaction was stirred at 10 °C for 17 hours.
  • the product was indicated present via UPLC analysis.
  • the solvent was removed under the reduced pressure and the residue was further purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10 ⁇ m column.
  • the mobile phase was acetonitrile/0.01% aqueous formic acid).
  • Racemic product N-(6- ethylbenzo[d]thiazol-2-yl)-3-methyl-N-(pyridin-3-ylmethyl)cyclobutanecarboxamide (80.0 mg, 0.22 mmol, 49 %) was afforded as a white solid.80 mg of this product was sent to chiral separation (the crude sample was dissolved in 35 mL methanol. AD 20*250 mm column. The mobile phase was 0.2% methanol/ammonia) to afford two isomers.
  • Example 148 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-2- yl)methyl]cyclohexanecarboxamide Step 1: Preparation of 6-fluoro-N-[(1H-imidazol-2-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of 6-fluoro-1,3-benzothiazol-2-amine 200 mg, 1.2 mmol
  • 1H- imidazole-2-carbaldehyde 113 mg, 1.2 mmol
  • dry toluene (12 mL) was refluxed at 120 °C with activated 4 ⁇ molecular sieves under N2 for 24 hours.
  • the reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry ethyl alcohol (12 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate and concentrated under reduced pressure.
  • the product was purified through flash column chromatography on a 24 gram silica gel column using a gradient of 0- 100% ethyl acetate in hexanes.
  • Step 2 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-2- yl)methyl]cyclohexanecarboxamide
  • 6-fluoro-N-[(1H-imidazol-2-yl)methyl]-1,3-benzothiazol-2-amine 199 mg, 0.80 mmol
  • triethylamine 0.17 mL, 1.2 mmol
  • cyclohexanecarbonyl chloride 174 mg, 1.2 mmol
  • Step 1 Preparation of 3-[(1H-imidazol-5-yl)methyl]-3-(6-methoxy-1,3-benzothiazol-2-yl)-1- phenylurea
  • Step 1 Preparation of 6-methoxy-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
  • 6-methoxybenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 ⁇ molecular sieves.
  • Step 2 Preparation of 3-[(1H-imidazol-5-yl)methyl]-3-(6-methoxy-1,3-benzothiazol-2-yl)-1-phenylurea
  • triphosgene 113 mg, 0.38 mmol
  • dichloromethane 10 mL
  • N-((1H-imidazol-5-yl)methyl)-6-methoxybenzo[d]thiazol-2-amine 200 mg, 0.77 mmol
  • pyridine 182 mg, 2.3 mmol
  • Step 2 Preparation of tert-butyl 4-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3- yl)methyl]carbamoyl]piperazine-1-carboxylate
  • triphosgene 113 mg, 0.38 mmol
  • dichloromethane 10 mL
  • N-((1H-imidazol-5-yl)methyl)-6-ethylbenzo[d]thiazol-2-amine 200 mg, 0.77 mmol
  • pyridine 182 mg, 2.3 mmol
  • Step 1 Preparation of 4-acetyl-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]piperazine-1-carboxamide
  • Step 1 Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
  • nicotinaldehyde 2.70 g, 25 mmol
  • 6-ethylbenzo[d]thiazol-2-amine 3.00 g, 17 mmol
  • toluene 60 mL
  • Step 2 Preparation of 4-acetyl-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperazine-1- carboxamide
  • triphosgene 113 mg, 0.38 mmol
  • dichloromethane 10 mL
  • N-((1H-imidazol-5-yl)methyl)-6-ethylbenzo[d]thiazol-2-amine 200 mg, 0.77 mmol
  • pyridine 182 mg, 2.3 mmol
  • Step 1 Preparation of N-(1,3-benzothiazol-2-yl)-4-cyano-N-[(1H-imidazol-5- yl)methyl]benzamide
  • Step 1 Preparation of N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine
  • 1-trityl-1H-imidazole-4-carbaldehyde (2.50 g, 7.4 mmol) in toluene (20 mL) was added dried 4 ⁇ molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours.
  • Step 2 Preparation of N-(benzo[d]thiazol-2-yl)-4-cyano-N-((1-trityl-1H-imidazol-5-yl)methyl)benzamide
  • N-((1-trityl-1H-imidazol-5-yl)methyl)benzo[d]thiazol-2-amine 400 mg, 0.85 mmol
  • 4- cyanobenzoic acid 125 mg, 0.85 mmol
  • N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (485 mg, 1.3 mmol) and N,N- diisopropylethylamine (219 mg, 1.7 mmol) in tetrahydrofuran (20 mL) was stirred at 70 °C for 16 hours.
  • Step 3 Preparation of N-(1,3-benzothiazol-2-yl)-4-cyano-N-[(1H-imidazol-5-yl)methyl]benzamide
  • dichloromethane 5 mL
  • trifluoroacetic acid 2 mL
  • the reaction was stirred at room temperature for 3 hours.
  • the product was indicated present via UPLC analysis.
  • the reaction was concentrated under reduced pressure and quenched with a solution of 7 M ammonia in methanol until pH 8.
  • Step 1 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-4-methyl-N-[(pyridin-3- yl)methyl]cyclohexane-1-carboxamide
  • Step 1 Preparation of 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of pyridine-3-carbaldehyde (0.63 mL, 6.7 mmol) and 6-ethyl-1,3-benzothiazol-2-amine (1.20 g, 6.7 mmol) in dry toluene (34 mL) was stirred at 105°C with activated 4 ⁇ molecular sieves under nitrogen overnight.
  • the reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (1.27 g, 34 mmol) in dry ethyl alcohol (34 mL). The reaction mixture was stirred at 70 °C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with ethyl acetate. The filtrate was concentrated then diluted with dichloromethane and filtered again. The filtrate was purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane through a 40 g column of silica gel.
  • Step 2 Preparation of 4-methylcyclohexane-1-carbonyl chloride
  • 4-methylcyclohexane-1-carboxylic acid 300 mg, 2.1 mmol
  • dichloromethane 11 mL
  • oxalyl chloride 0.89 mL, 11 mmol
  • N,N-dimethylformamide were added as a catalyst and the reaction bubbled.
  • the reaction was then heated to 40 °C and stirred overnight.
  • the product was indicated present via crude NMR analysis.
  • the reaction was cooled to room temperature and concentrated under reduced pressure. Crude product was indicated pure enough to continue to next step via NMR and assumed 100% yield.
  • Step 3 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-4-methyl-N-[(pyridin-3-yl)methyl]cyclohexane-1- carboxamide
  • 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine 400 mg, 1.5 mmol
  • tetrahydrofuran 7.4 mL
  • Step 1 Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(4-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 6-methoxy-N-[(4-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
  • 4-methoxypyridine-3-carbaldehyde 150 mg, 1.1 mmol
  • 6- methoxy-1,3-benzothiazol-2-amine 200 mg, 1.1 mmol
  • dry toluene 11 mL
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (95.7 mg, 2.5 mmol) in dry ethyl alcohol (11 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(4-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide
  • 6-methoxy-N-[(4-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine 255 mg, 0.85 mmol
  • triethylamine (0.17 mL, 1.3 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C.
  • cyclohexanecarbonyl chloride (368 mg, 2.6 mmol) was added dropwise with stirring.
  • Example 155 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(5-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide Step 1: Preparation of 6-ethyl-N-[(5-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of 5-methoxypyridine-3-carbaldehyde 153 mg, 1.1 mmol
  • 6- ethyl-1,3-benzothiazol-2-amine 200 mg, 1.1 mmol
  • dry toluene 11 mL
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(5-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide
  • 6-ethyl-N-[(5-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (271 mg, 0.90 mmol) and triethylamine (0.19 mL, 1.4 mmol) were dissolved in dimethylformamide (1.1 mL) and cooled to 0 °C.
  • cyclohexanecarbonyl chloride (197 mg, 1.4 mmol) was added dropwise with stirring.
  • the reaction was warmed to room temperature and stirred overnight.
  • the product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-25% ethyl acetate in hexanes.
  • Example 156 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(6-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide Step 1: Preparation of 6-ethyl-N-[(6-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of 6-methoxypyridine-3-carbaldehyde 153 mg, 1.1 mmol
  • 6- ethyl-1,3-benzothiazol-2-amine 200 mg, 1.1 mmol
  • dry toluene 11 mL
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane.
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(6-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide
  • 6-ethyl-N-[(6-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (332 mg, 1.1 mmol) and triethylamine (0.23 mL, 1.7 mmol) were dissolved in dimethylformamide (1.4 mL) and cooled to 0 °C.
  • cyclohexanecarbonyl chloride (241 mg, 1.7 mmol) was added dropwise with stirring.
  • Example 157 Preparation of 1-cyclopropanecarbonyl-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin- 3-yl)methyl]piperidine-4-carboxamide
  • 1-cyclopropanecarbonyl-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]piperidine-4-carboxamide To a solution of N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)piperidine-4-carboxamide (50.0 mg, 0.13 mmol) in dichloromethane (3 mL) was added triethylamine (26.0 mg, 0.26 mmol) and cyclopropanecarbonyl chloride (16.0 mg, 0.18 mmol).
  • the reaction was stirred at room temperature for 2 hours.
  • the product was indicated present via UPLC analysis.
  • the reaction was diluted with water (3 mL), then extracted with 3x 15 mL portions of dichloromethane. The organic layers were separated, pooled, filtered over sodium sulfate, and concentrated under reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston pHlex ODS 10um 21.2 ⁇ 250mm 120A. The mobile phase was acetonitrile/0.1% ammonium bicarbonate).
  • the reaction was stirred at room temperature for 2 hours.
  • the product was indicated present via UPLC analysis.
  • the reaction was diluted with water (3 mL), then extracted with 3x 15 mL portions of dichloromethane. The organic layers were separated, pooled, filtered over sodium sulfate, and concentrated under reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston pHlex ODS 10um 21.2 ⁇ 250mm 120A. The mobile phase was acetonitrile/0.1% ammonium bicarbonate).
  • Example 159 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(2-methoxypyridin-4- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 6-ethyl-N-((2-methoxypyridin-4-yl)methyl)benzo[d]thiazol-2-amine
  • 2-methoxyisonicotinaldehyde 233 mg, 1.7 mmol
  • toluene 40 mL
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(2-methoxypyridin-4- yl)methyl]cyclohexanecarboxamide
  • 6-ethyl-N-((2-methoxypyridin-4-yl)methyl)benzo[d]thiazol-2-amine 150 mg, 0.50 mmol
  • N-dimethylformamide 5 mL
  • 60% w/w sodium hydride in oil 30.0 mg, 0.75 mmol
  • cyclohexanecarbonyl chloride 95.0 mg, 0.65 mmol
  • the product was indicated present via UPLC analysis.
  • the mixture was quenched with ice water, then extracted with ethyl acetate (2x 15 mL). The organic layers were pooled, washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure.
  • the residue was purified by prep-HPLC (the crude samples were dissolved in methanol before purification. Boston C1821*250mm 10 ⁇ m column. The mobile phase was acetonitrile/0.01% aqueous ammonium bicarbonate).
  • Example 160 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 5-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine
  • 5-fluorobenzo[d]thiazol-2-amine 50.0 mg, 0.30 mmol
  • 1-trityl-1H-imidazole-4- carbaldehyde 151 mg, 0.45 mmol
  • Step 2 Preparation of N-(5-Fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide
  • 5-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine 100 mg, 0.20 mmol
  • triethylamine 0.085 mL, 0.61 mmol
  • dichloromethane 8 mL
  • cyclohexanecarbonyl chloride 45.0 mg, 0.31 mmol
  • Step 3 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
  • N-(5-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide 78.0 mg, 0.13 mmol
  • dichloromethane 5 mL
  • trifluoroacetic acid 1.5 mL
  • Step 1 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 4-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine
  • 4-fluorobenzo[d]thiazol-2-amine 50.0 mg, 0.30 mmol
  • 1-trityl-1H-imidazole-4- carbaldehyde 151 mg, 0.45 mmol
  • Step 2 Preparation of N-(4-Fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide
  • 4-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine 100 mg, 0.20 mmol
  • triethylamine 0.085 mL, 0.61 mmol
  • dichloromethane 8 mL
  • cyclohexanecarbonyl chloride 45.0 mg, 0.31 mmol
  • Step 3 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
  • N-(4-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide 80.0 mg, 0.13 mmol
  • dichloromethane 5 mL
  • trifluoroacetic acid 1.5 mL
  • Step 1 Preparation of N-(7-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 7-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine
  • 7-fluorobenzo[d]thiazol-2-amine 50.0 mg, 0.30 mmol
  • 1-trityl-1H-imidazole-4- carbaldehyde 151 mg, 0.45 mmol
  • Step 2 Preparation of N-(7-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide
  • a solution of 7-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine 100 mg, 0.20 mmol
  • triethylamine 0.085 mL, 0.61 mmol
  • dichloromethane 8 mL
  • cyclohexanecarbonyl chloride 45.0 mg, 0.31 mmol
  • Step 3 Preparation of N-(7-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
  • N-(7-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide 80.0 mg, 0.13 mmol
  • dichloromethane 5 mL
  • trifluoroacetic acid 1.5 mL
  • Step 1 Preparation of phenyl N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2- yl)carbamate
  • Step 1 Preparation of 6-methoxy-N-((1-trityl-1H-imidazol-5-yl)methyl)benzo[d]thiazol-2-amine
  • 6-methoxybenzo[d]thiazol-2-amine 300 mg, 1.7 mmol
  • 1-trityl-1H-imidazole-4- carbaldehyde 563 mg, 1.7 mmol
  • toluene 40 mL
  • Step 2 Preparation of phenyl (6-methoxybenzo[d]thiazol-2-yl)((1-trityl-1H-imidazol-5-yl)methyl)carbamate
  • 6-methoxy-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine 100 mg, 0.20 mmol
  • N,N-dimethylformamide 5 mL
  • 60% w/w sodium hydride in oil (12.0 mg, 0.30 mmol
  • phenyl carbonochloridate 41.0 mg, 0.26 mmol
  • Step 3 Preparation of phenyl N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2-yl)carbamate
  • dichloromethane 5 mL
  • trifluoroacetic acid 2 mL
  • the product was indicated present via UPLC analysis.
  • the resulting mixture was concentrated and quenched with a solution of 7 M ammonia in methanol, until pH 8.
  • Example 165 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-1- propanoylpiperidine-4-carboxamide
  • Step 1 Preparation of 1-propionylpiperidine-4-carboxylic acid
  • N,N-diisopropylethylamine 593 mg, 4.6 mmol
  • propionyl chloride 275 mg, 3.0 mmol
  • the mixture was purified directly by reversed-phase chromatography (mobile phase was 15% acetonitrile/0.01% aqueous formic acid) to offer 1-propionylpiperidine-4-carboxylic acid (200 mg, 1.1 mmol, 48 %) as a white solid.
  • Step 2 Preparation of N-(6-fluorobenzo[d]thiazol-2-yl)-1-propionyl-N-((1-trityl-1H-imidazol-5- yl)methyl)piperidine-4-carboxamide
  • the product was indicated present via UPLC analysis.
  • the reaction was quenched with ice water, then extracted with ethyl acetate (2x 15 mL). The organic layers were pooled, washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by flash column chromatography through silica gel using a gradient of 0-25% ethyl acetate in petroleum ether.
  • Step 3 Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-1- propanoylpiperidine-4-carboxamide
  • N-(6-fluorobenzo[d]thiazol-2-yl)-1-propionyl-N-((1-trityl-1H-imidazol-5- yl)methyl)piperidine-4-carboxamide 120 mg, 0.18 mmol
  • dichloromethane 5 mL
  • trifluoroacetic acid 2 mL
  • Example 166 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-4- yl)methyl]propenamide
  • Step 1 Preparation of 5-fluoro-N-(pyridin-4-ylmethyl)benzo[d]thiazol-2-amine
  • isonicotinaldehyde 321 mg, 3.0 mmol
  • Step 2 Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-4-yl)methyl]propanamide
  • 5-fluoro-N-(pyridin-4-ylmethyl)benzo[d]thiazol-2-amine 500 mg, 1.9 mmol
  • pyridine 0.5 mL
  • N,N-dimethylformamide 2 mL
  • dichloromethane 5 mL
  • Step 1 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-3- phenoxypropanamide
  • Step 1 Preparation of 4-fluoro-N-[(1H-imidazol-4-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of 1H-imidazole-4-carbaldehyde 113 mg, 1.2 mmol
  • 4-fluoro-1,3- benzothiazol-2-amine 200 mg, 1.2 mmol
  • dry toluene (12 mL) was refluxed at 120 °C with activated 4 ⁇ molecular sieves under N2 for 24 hours.
  • reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry ethyl alcohol (12 mL).
  • the resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature.
  • the product was indicated present via UPLC analysis.
  • the reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate.
  • the product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-20% methanol in ethyl acetate.
  • Step 2 Preparation of 3-phenoxypropanoyl chloride
  • 3-phenoxypropanoic acid 500 mg, 3.0 mmol
  • dichloromethane 6 mL
  • oxalyl chloride (1.26 mL, 15 mmol) was added dropwise, followed by a few drops of N,N-dimethylformamide.
  • the reaction bubbled.
  • the reaction was heated to 47°C and stir for 2 hours.
  • the reaction was then cool to room temperature and concentrated under reduced pressure. Crude product was indicated pure enough to continue to next step via NMR.
  • Step 3 Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-3- phenoxypropanamide
  • 4-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine 104 mg, 0.42 mmol
  • triethylamine 0.087 mL, 0.63 mmol
  • Step 1 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-2- yl)methyl]cyclohexanecarboxamide
  • Step 1 Preparation of 6-ethyl-N-[(1H-imidazol-2-yl)methyl]-1,3-benzothiazol-2-amine
  • a solution of 1H-imidazole-2-carbaldehyde 188 mg, 2.0 mmol
  • 6-ethyl-1,3- benzothiazol-2-amine 350 mg, 2.0 mmol
  • dry toluene (10 mL) was stirred at 100 °C with activated 4 ⁇ molecular sieves under nitrogen for 24 hours.
  • the reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (222 mg, 5.9 mmol) in dry ethyl alcohol (10 mL). The reaction mixture was stirred at 65°C for 1 hour and then cooled to room temperature. The reaction was filtered over a bed of Celite and washed with ethyl acetate (2x 20 mL). The filtrate was diluted with ethyl acetate and washed with sodium bicarbonate. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated.
  • Step 2 Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-2- yl)methyl]cyclohexanecarboxamide
  • 6-ethyl-N-[(1H-imidazol-2-yl)methyl]-1,3-benzothiazol-2-amine (324 mg, 1.3 mmol) was dissolved in tetrahydrofuran (6 mL) and cooled to 0 °C.
  • the mobile phase was acetonitrile/0.1% ammonium bicarbonate) to afford the desired compound N-(6- ethylbenzo[d]thiazol-2-yl)-1-(2-(1-methylcyclopropyl)acetyl)-N-(pyridin-3- ylmethyl)piperidine-4-carboxamide (17.5 mg, 0.040 mmol, 28 %) as a white solid.

Abstract

The present invention features compounds useful in the treatment of neurological disorders. The compounds of the invention, alone or in combination with other pharmaceutically active agents, can be used for treating or preventing neurological disorders.

Description

BENZOTHIAZOLE COMPOUNDS AND USES THEREOF Background An incomplete understanding of the molecular perturbations that cause disease, as well as a limited arsenal of robust model systems, has contributed to a failure to generate successful disease-modifying therapies against common and progressive neurological disorders, such as ALS and FTD. Progress is being made on many fronts to find agents that can arrest the progress of these disorders. However, the present therapies for most, if not all, of these diseases provide very little relief. Accordingly, a need exists to develop therapies that can alter the course of neurodegenerative diseases. More generally, a need exists for better methods and compositions for the treatment of neurodegenerative diseases in order to improve the quality of the lives of those afflicted by such diseases. Summary TDP-43 is a nuclear DNA/RNA binding protein involved in RNA splicing. Under pathological cell stress, TDP-43 translocates to the cytoplasm and aggregates into stress granules. These phenotypes are hallmarks of degenerating motor neurons and are found in 97% of all ALS cases. The highly penetrant nature of this pathology indicates that TDP-43 is broadly involved in both familial and sporadic ALS. Additionally, TDP-43 mutations that promote aggregation are linked to higher risk of developing ALS, suggesting protein misfolding and aggregation act as drivers of toxicity. TDP-43 toxicity can be recapitulated in yeast models, where the protein induces a viability deficit and localizes to stress granules. The present inventors have discovered that the CYP51A1 inhibitors described herein are capable of reversing TDP-43 induced toxicity. Accordingly, the present invention describes such CYP51A1 inhibitors and methods of using these compounds for the treatment of disorders related to TDP-43 toxicity such as ALS. In an aspect, the invention features a compound, or a pharmaceutically acceptable salt thereof, having the structure:
Figure imgf000002_0001
Formula I wherein n is 0, 1, 2, 3, or 4; X is S or O; each R1 is independently halo, cyano, hydroxy, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, -SO2-optionally substituted C1-C6 alkyl, or -CO2-optionally substituted C1-C6 alkyl; R2 is optionally substituted C2-C9 heteroaryl; and R3 is optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C9 heterocyclyl, optionally substituted C1-C6 alkyl C3-C8 cycloalkyl, optionally substituted C1-C6 heteroalkyl C3-C8 cycloalkyl, optionally substituted C1-C6 alkyl C2-C9 heteroaryl, optionally substituted C1-C6 heteroalkyl C2-C9 heteroaryl, optionally substituted C1-C6 alkyl C2-C9 heterocyclyl, optionally substituted C1-C6 heteroalkyl C2-C9 heterocyclyl, optionally substituted C1-C6 alkyl C6-C10 aryl, or optionally substituted C1-C6 heteroalkyl C6-C10 aryl. In some embodiments, X is S. In some embodiments, X is O. In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, the compound has the structure:
Figure imgf000003_0001
. Formula Ia In some embodiments, the compound has the structure:
Figure imgf000003_0002
. Formula Ib In some embodiments, the compound has the structure:
Figure imgf000003_0003
. Formula Ic In some embodiments, the compound has the structure:
Figure imgf000003_0004
. Formula Id In some embodiments, at least one R1 is halo (e.g., fluoro, chloro, or bromo). In some embodiments, at least one R1 is cyano. In some embodiments, at least one R1 is hydroxy. In some embodiments, at least one R1 is optionally substituted C1-C6 alkyl (e.g., methyl, ethyl, trifluoromethyl, or hydroxymethyl). In some embodiments, at least one R1 is optionally substituted C1-C6 heteroalkyl (e.g., ethoxy or trifluoromethoxy). In some embodiments, at least one R1 is -SO2-optionally substituted C1-C6 alkyl (e.g., -SO2-methyl). In some embodiments, at least one R1 is -CO2-optionally substituted C1-C6 alkyl (e.g., -CO2-methyl or -CO2-ethyl). In some embodiments, R2 is a 5-membered optionally substituted C2-C9 heteroaryl (e.g.,
Figure imgf000003_0005
some embodiments, R2 is a 6-membered optionally substituted C2-C9 heteroaryl (e.g., , ,
Figure imgf000004_0001
In some embodiments, R3 is optionally substituted C3-C8 cycloalkyl (e.g., cyclohexyl). In some embodiments, R3 is optionally substituted C2-C9 heterocyclyl (e.g.,
Figure imgf000004_0002
, , ,
Figure imgf000004_0003
, , or , wherein R4 is -C(O)R5; R5 is optionally substituted C1- C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C1-C6 alkyl C3-C8 cycloalkyl, or optionally substituted C1-C6 heteroalkyl C3-C8 cycloalkyl). In some embodiments, R3 is optionally substituted C6-C10 aryl (e.g., phenyl, 4-cyano-phenyl, or 4-fluoro-phenyl). In an aspect, the invention features a compound, or a pharmaceutically acceptable salt thereof, having the structure of any one of compounds 1-273 in Table 1.
Table 1. Compounds of the Invention
Figure imgf000005_0001
Figure imgf000006_0001
Figure imgf000007_0001
Figure imgf000008_0001
Figure imgf000009_0001
Figure imgf000010_0001
Figure imgf000011_0001
Figure imgf000012_0001
Figure imgf000013_0001
Figure imgf000014_0001
Figure imgf000015_0001
Figure imgf000016_0001
Figure imgf000017_0001
Figure imgf000018_0001
Figure imgf000019_0001
Figure imgf000020_0001
Figure imgf000021_0001
Figure imgf000022_0001
Figure imgf000023_0001
Figure imgf000024_0001
In an aspect, the invention features a pharmaceutical composition comprising any of the foregoing compounds and a pharmaceutically acceptable excipient. In an aspect, the invention features a method of treating a neurological disorder (e.g., frontotemporal dementia (FTLD-TDP), chronic traumatic encephalopathy, ALS, Alzheimer’s disease, limbic-predominant age-related TDP-42 encephalopathy (LATE), or frontotemporal lobar degeneration) in a subject in need thereof. This method includes administering an effective amount of any of the foregoing compounds or pharmaceutical compositions. In an aspect, the invention features a method of inhibiting toxicity in a cell (e.g., mammalian neural cell) related to a protein (e.g., TDP-43). This method includes administering an effective amount of any of the foregoing compounds or pharmaceutical compositions. In an aspect, the invention features a method of treating a CYP51A1-associated disorder (e.g., FTLD-TDP, chronic traumatic encephalopathy, ALS, Alzheimer’s disease, LATE, or frontotemporal lobar degeneration) in a subject in need thereof. This method includes administering an effective amount of any of the foregoing compounds pharmaceutical compositions. In an aspect, the invention features a method of inhibiting CYP51A1. This method includes contacting a cell with an effective amount of any of the foregoing compounds or pharmaceutical compositions. In another aspect, the invention features a method of treating a neurological disorder in a patient, such as a human patient, identified as likely to benefit from treatment with a CYP51A1 inhibitor on the basis of TDP-43 aggregation. In this aspect, the method may include (i) determining that the patient exhibits, or is prone to develop, TDP-43 aggregation, and (ii) providing to the patient a therapeutically effective amount of a CYP51A1 inhibitor. In some embodiments, the patient has previously been determined to exhibit, or to be prone to developing, TDP-43 aggregation, and the method includes providing to the patient a therapeutically effective amount of a CYP51A1 inhibitor. The susceptibility of the patient to developing TDP-43 aggregation may be determined, e.g., by determining whether the patient expresses a mutant isoform of TDP-43 containing a mutation that is associated with TDP-43 aggregation and toxicity, such as a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D. This may be performed, for example, by determining the amino acid sequence of a TDP-43 isoform isolated from a sample obtained from the patient or by determining the nucleic acid sequence of a TDP-43 gene isolated from a sample obtained from the patient. In some embodiments, the method includes the step of obtaining the sample from the patient. In an additional aspect, the invention features a method of treating a neurological disorder in a patient, such as a human patient, identified as likely to benefit from treatment with a CYP51A1 inhibitor on the basis of TDP-43 expression. In this aspect, the method includes (i) determining that the patient expresses a mutant form of TDP-43 having a mutation associated with TDP-43 aggregation (e.g., a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D), and (ii) providing to the patient a therapeutically effective amount of a CYP51A1 inhibitor. In some embodiments, the patient has previously been determined to express a mutant form of TDP-43 having a mutation associated with TDP- 43 aggregation, such as a Q331K, M337V, Q343R, N345K, R361S, or N390D mutation, and the method includes providing to the patient a therapeutically effective amount of a CYP51A1 inhibitor. In another aspect, the invention features a method of determining whether a patient (e.g., a human patient) having a neurological disorder is likely to benefit from treatment with a CYP51A1 inhibitor by (i) determining whether the patient exhibits, or is prone to develop, TDP-43 aggregation and (ii) identifying the patient as likely to benefit from treatment with a CYP51A1 inhibitor if the patient exhibits, or is prone to develop, TDP-43 aggregation. In some embodiments, the method further includes the step of (iii) informing the patient whether he or she is likely to benefit from treatment with a CYP51A1 inhibitor. The susceptibility of the patient to developing TDP-43 aggregation may be determined, e.g., by determining whether the patient expresses a mutant isoform of TDP-43 containing a mutation that is associated with TDP-43 aggregation and toxicity, such as a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D. This may be performed, for example, by determining the amino acid sequence of a TDP-43 isoform isolated from a sample obtained from the patient or by determining the nucleic acid sequence of a TDP-43 gene isolated from a sample obtained from the patient. In some embodiments, the method includes the step of obtaining the sample from the patient. In another aspect, the invention features a method of determining whether a patient (e.g., a human patient) having a neurological disorder is likely to benefit from treatment with a CYP51A1 inhibitor by (i) determining whether the patient expresses a TDP-43 mutant having a mutation associated with TDP-43 aggregation (e.g., a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D) and (ii) identifying the patient as likely to benefit from treatment with a CYP51A1 inhibitor if the patient expresses a TDP-43 mutant. In some embodiments, the method further includes the step of (iii) informing the patient whether he or she is likely to benefit from treatment with a CYP51A1 inhibitor. The TDP-43 isoform expressed by the patient may be assessed, for example, by isolated TDP-43 protein from a sample obtained from the patient and sequencing the protein using molecular biology techniques described herein or known in the art. In some embodiments, the TDP-43 isoform expressed by the patient is determined by analyzing the patient’s genotype at the TDP-43 locus, for example, by sequencing the TDP-43 gene in a sample obtained from the patient. In some embodiments, the method includes the step of obtaining the sample from the patient. In some embodiments of any of the above aspects, the CYP51A1 inhibitor is provided to the patient by administration of the CYP51A1 inhibitor to the patient. In some embodiments, the CYP51A1 inhibitor is provided to the patient by administration of a prodrug that is converted in vivo to the CYP51A1 inhibitor. In some embodiments of any of the above aspects, the neurological disorder is a neuromuscular disorder, such as a neuromuscular disorder selected from amyotrophic lateral sclerosis, congenital myasthenic syndrome, congenital myopathy, cramp fasciculation syndrome, Duchenne muscular dystrophy, glycogen storage disease type II, hereditary spastic paraplegia, inclusion body myositis, Isaac's Syndrome, Kearns-Sayre syndrome, Lambert–Eaton myasthenic syndrome, mitochondrial myopathy, muscular dystrophy, myasthenia gravis, myotonic dystrophy, peripheral neuropathy, spinal and bulbar muscular atrophy, spinal muscular atrophy, Stiff person syndrome, Troyer syndrome, and Guillain– Barré syndrome. In some embodiments, the neurological disorder is amyotrophic lateral sclerosis. In some embodiments of any of the above aspects, the neurological disorder is selected from frontotemporal degeneration (also referred to as frontotemporal lobar degeneration and frontotemporal dementia), Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’s disease, Inclusion body myopathy with early-onset Paget disease and frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease, and hereditary inclusion body myopathy. In some embodiments, the neurological disorder is amyotrophic lateral sclerosis, and following administration of the CYP51A1 inhibitor to the patient, the patient exhibits one or more, or all, of the following responses: (i) an improvement in condition as assessed using the amyotrophic lateral sclerosis functional rating scale (ALSFRS) or the revised ALSFRS (ALSFRS-R), such as an improvement in the patient’s ALSFRS or ALSFRS-R score within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an improvement in the patient’s ALSFRS or ALSFRS-R score within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the patient); (ii) an increase in slow vital capacity, such as an increase in the patient’s slow vital capacity within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an increase in the patient’s slow vital capacity within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the patient); (iii) a reduction in decremental responses exhibited by the patient upon repetitive nerve stimulation, such as a reduction that is observed within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., a reduction that is observed within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the patient); (iv) an improvement in muscle strength, as assessed, for example, by way of the Medical Research Council muscle testing scale (as described, e.g., in Jagtap et al., Ann. Indian. Acad. Neurol. 17:336-339 (2014), the disclosure of which is incorporated herein by reference as it pertains to measuring patient response to neurological disease treatment), such as an improvement that is observed within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an improvement that is observed within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the patient); (v) an improvement in quality of life, as assessed, for example, using the amyotrophic lateral sclerosis-specific quality of life (ALS-specific QOL) questionnaire, such as an improvement in the patient’s quality of life that is observed within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an improvement in the subject’s quality of life that is observed within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the patient); (vi) a decrease in the frequency and/or severity of muscle cramps, such as a decrease in cramp frequency and/or severity within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., a decrease in cramp frequency and/or severity within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the patient); and/or (vii) a decrease in TDP-43 aggregation, such as a decrease in TDP-43 aggregation within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., a decrease in TDP- 43 aggregation within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the patient. Chemical Terms It is to be understood that the terminology employed herein is for the purpose of describing particular embodiments and is not intended to be limiting. Those skilled in the art will appreciate that certain compounds described herein can exist in one or more different isomeric (e.g., stereoisomers, geometric isomers, tautomers) and/or isotopic (e.g., in which one or more atoms has been substituted with a different isotope of the atom, such as hydrogen substituted for deuterium) forms. Unless otherwise indicated or clear from context, a depicted structure can be understood to represent any such isomeric or isotopic form, individually or in combination. In some embodiments, one or more compounds depicted herein may exist in different tautomeric forms. As will be clear from context, unless explicitly excluded, references to such compounds encompass all such tautomeric forms. In some embodiments, tautomeric forms result from the swapping of a single bond with an adjacent double bond and the concomitant migration of a proton. In certain embodiments, a tautomeric form may be a prototropic tautomer, which is an isomeric protonation states having the same empirical formula and total charge as a reference form. Examples of moieties with prototropic tautomeric forms are ketone – enol pairs, amide – imidic acid pairs, lactam – lactim pairs, amide – imidic acid pairs, enamine – imine pairs, and annular forms where a proton can occupy two or more positions of a heterocyclic system, such as, 1H- and 3H-imidazole, 1H-, 2H- and 4H- 1,2,4-triazole, 1H- and 2H- isoindole, and 1H- and 2H-pyrazole. In some embodiments, tautomeric forms can be in equilibrium or sterically locked into one form by appropriate substitution. In certain embodiments, tautomeric forms result from acetal interconversion, e.g., the interconversion illustrated in the scheme below:
Figure imgf000029_0001
. Those skilled in the art will appreciate that, in some embodiments, isotopes of compounds described herein may be prepared and/or utilized in accordance with the present invention. “Isotopes” refers to atoms having the same atomic number but different mass numbers resulting from a different number of neutrons in the nuclei. For example, isotopes of hydrogen include tritium and deuterium. In some embodiments, an isotopic substitution (e.g., substitution of hydrogen with deuterium) may alter the physiciochemical properties of the molecules, such as metabolism and/or the rate of racemization of a chiral center. As is known in the art, many chemical entities (in particular many organic molecules and/or many small molecules) can adopt a variety of different solid forms such as, for example, amorphous forms and/or crystalline forms (e.g., polymorphs, hydrates, solvates, etc). In some embodiments, such entities may be utilized in any form, including in any solid form. In some embodiments, such entities are utilized in a particular form, for example in a particular solid form. In some embodiments, compounds described and/or depicted herein may be provided and/or utilized in salt form. In certain embodiments, compounds described and/or depicted herein may be provided and/or utilized in hydrate or solvate form. At various places in the present specification, substituents of compounds of the present disclosure are disclosed in groups or in ranges. It is specifically intended that the present disclosure include each and every individual subcombination of the members of such groups and ranges. For example, the term “C1-C6 alkyl” is specifically intended to individually disclose methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl. Furthermore, where a compound includes a plurality of positions at which substitutes are disclosed in groups or in ranges, unless otherwise indicated, the present disclosure is intended to cover individual compounds and groups of compounds (e.g., genera and subgenera) containing each and every individual subcombination of members at each position. Herein a phrase of the form “optionally substituted X” (e.g., optionally substituted alkyl) is intended to be equivalent to “X, wherein X is optionally substituted” (e.g., “alkyl, wherein said alkyl is optionally substituted”). It is not intended to mean that the feature “X” (e.g. alkyl) per se is optional. The term “acyl,” as used herein, represents a hydrogen or an alkyl group, as defined herein that is attached to a parent molecular group through a carbonyl group, as defined herein, and is exemplified by formyl (i.e., a carboxyaldehyde group), acetyl, trifluoroacetyl, propionyl, and butanoyl. Exemplary unsubstituted acyl groups include from 1 to 6, from 1 to 11, or from 1 to 21 carbons. The term “alkyl,” as used herein, refers to a branched or straight-chain monovalent saturated aliphatic hydrocarbon radical of 1 to 20 carbon atoms (e.g., 1 to 16 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms). An alkylene is a divalent alkyl group. The term “alkenyl,” as used herein, alone or in combination with other groups, refers to a straight-chain or branched hydrocarbon residue having a carbon-carbon double bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6, or 2 carbon atoms). The term “alkynyl,” as used herein, alone or in combination with other groups, refers to a straight-chain or branched hydrocarbon residue having a carbon-carbon triple bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6, or 2 carbon atoms). The term “amino,” as used herein, represents -N(RN1)2, wherein each RN1 is, independently, H, OH, NO2, N(RN2)2, SO2ORN2, SO2RN2, SORN2, an N-protecting group, alkyl, alkoxy, aryl, arylalkyl, cycloalkyl, acyl (e.g., acetyl, trifluoroacetyl, or others described herein), wherein each of these recited RN1 groups can be optionally substituted; or two RN1 combine to form an alkylene or heteroalkylene, and wherein each RN2 is, independently, H, alkyl, or aryl. The amino groups of the invention can be an unsubstituted amino (i.e., -NH2) or a substituted amino (i.e., -N(RN1)2). The term “aryl,” as used herein, refers to an aromatic mono- or polycarbocyclic radical of 6 to 12 carbon atoms having at least one aromatic ring. Examples of such groups include, but are not limited to, phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, 1,2-dihydronaphthyl, indanyl, and 1H-indenyl. The term “arylalkyl,” as used herein, represents an alkyl group substituted with an aryl group. Exemplary unsubstituted arylalkyl groups are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as C1-C6 alkyl C6-10 aryl, C1-C10 alkyl C6-10 aryl, or C1-C20 alkyl C6-10 aryl), such as, benzyl and phenethyl. In some embodiments, the akyl and the aryl each can be further substituted with 1, 2, 3, or 4 substituent groups as defined herein for the respective groups. The term “azido,” as used herein, represents a -N3 group. The term “cyano,” as used herein, represents a CN group. The terms “carbocyclyl,” as used herein, refer to a non-aromatic C3-C12 monocyclic, bicyclic, or tricyclic structure in which the rings are formed by carbon atoms. Carbocyclyl structures include cycloalkyl groups and unsaturated carbocyclyl radicals. The term “cycloalkyl,” as used herein, refers to a saturated, non-aromatic, monovalent mono- or polycarbocyclic radical of three to ten, preferably three to six carbon atoms. This term is further exemplified by radicals such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, and adamantyl. The term “halo,” as used herein, means a fluorine (fluoro), chlorine (chloro), bromine (bromo), or iodine (iodo) radical. The term “heteroalkyl,” as used herein, refers to an alkyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkyl group can be further substituted with 1, 2, 3, or 4 substituent groups as described herein for alkyl groups. Examples of heteroalkyl groups are an “alkoxy” which, as used herein, refers alkyl-O- (e.g., methoxy and ethoxy). A heteroalkylene is a divalent heteroalkyl group. The term “heteroalkenyl,” as used herein, refers to an alkenyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkenyl group can be further substituted with 1, 2, 3, or 4 substituent groups as described herein for alkenyl groups. Examples of heteroalkenyl groups are an “alkenoxy” which, as used herein, refers alkenyl-O-. A heteroalkenylene is a divalent heteroalkenyl group. The term “heteroalkynyl,” as used herein, refers to an alkynyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkynyl group can be further substituted with 1, 2, 3, or 4 substituent groups as described herein for alkynyl groups. Examples of heteroalkynyl groups are an “alkynoxy” which, as used herein, refers alkynyl-O-. A heteroalkynylene is a divalent heteroalkynyl group. The term “heteroaryl,” as used herein, refers to an aromatic mono- or polycyclic radical of 5 to 12 atoms having at least one aromatic ring containing one, two, or three ring heteroatoms selected from N, O, and S, with the remaining ring atoms being C. One or two ring carbon atoms of the heteroaryl group may be replaced with a carbonyl group. Examples of heteroaryl groups are pyridyl, pyrazoyl, benzooxazolyl, benzoimidazolyl, benzothiazolyl, imidazolyl, oxaxolyl, and thiazolyl. The term “heteroarylalkyl,” as used herein, represents an alkyl group substituted with a heteroaryl group. Exemplary unsubstituted heteroarylalkyl groups are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as C1-C6 alkyl C2-C9 heteroaryl, C1-C10 alkyl C2-C9 heteroaryl, or C1-C20 alkyl C2-C9 heteroaryl). In some embodiments, the akyl and the heteroaryl each can be further substituted with 1, 2, 3, or 4 substituent groups as defined herein for the respective groups. The term “heterocyclyl,” as used herein, denotes a mono- or polycyclic radical having 3 to 12 atoms having at least one ring containing one, two, three, or four ring heteroatoms selected from N, O or S and no aromatic ring containing any N, O, or S atoms. Examples of heterocyclyl groups include, but are not limited to, morpholinyl, thiomorpholinyl, furyl, piperazinyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydropyranyl, tetrahydrofuranyl, and 1,3-dioxanyl. The term “heterocyclylalkyl,” as used herein, represents an alkyl group substituted with a heterocyclyl group. Exemplary unsubstituted heterocyclylalkyl groups are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as C1-C6 alkyl C2-C9 heterocyclyl, C1-C10 alkyl C2-C9 heterocyclyl, or C1-C20 alkyl C2-C9 heterocyclyl). In some embodiments, the akyl and the heterocyclyl each can be further substituted with 1, 2, 3, or 4 substituent groups as defined herein for the respective groups. The term “hydroxyl,” as used herein, represents an -OH group. The term “N-protecting group,” as used herein, represents those groups intended to protect an amino group against undesirable reactions during synthetic procedures. Commonly used N-protecting groups are disclosed in Greene, “Protective Groups in Organic Synthesis,” 3rd Edition (John Wiley & Sons, New York, 1999). N-protecting groups include acyl, aryloyl, or carbamyl groups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, and chiral auxiliaries such as protected or unprotected D, L or D, L-amino acids such as alanine, leucine, and phenylalanine; sulfonyl-containing groups such as benzenesulfonyl, and p-toluenesulfonyl; carbamate forming groups such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1-(p-biphenylyl)-1-methylethoxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxy carbonyl, t-butyloxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2,-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxy carbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, and phenylthiocarbonyl, arylalkyl groups such as benzyl, triphenylmethyl, and benzyloxymethyl, and silyl groups, such as trimethylsilyl. Preferred N-protecting groups are alloc, formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, alanyl, phenylsulfonyl, benzyl, t-butyloxycarbonyl (Boc), and benzyloxycarbonyl (Cbz). The term “nitro,” as used herein, represents an NO2 group. The term “thiol,” as used herein, represents an -SH group. The alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl (e.g., cycloalkyl), aryl, heteroaryl, and heterocyclyl groups may be substituted or unsubstituted. When substituted, there will generally be 1 to 4 substituents present, unless otherwise specified. Substituents include, for example: aryl (e.g., substituted and unsubstituted phenyl), carbocyclyl (e.g., substituted and unsubstituted cycloalkyl), halo (e.g., fluoro), hydroxyl, oxo, heteroalkyl (e.g., substituted and unsubstituted methoxy, ethoxy, or thioalkoxy), heteroaryl, heterocyclyl, amino (e.g., NH2 or mono- or dialkyl amino), azido, cyano, nitro, or thiol. Aryl, carbocyclyl (e.g., cycloalkyl), heteroaryl, and heterocyclyl groups may also be substituted with alkyl (unsubstituted and substituted such as arylalkyl (e.g., substituted and unsubstituted benzyl)). Compounds of the invention can have one or more asymmetric carbon atoms and can exist in the form of optically pure enantiomers, mixtures of enantiomers such as, for example, racemates, optically pure diastereoisomers, mixtures of diastereoisomers, diastereoisomeric racemates or mixtures of diastereoisomeric racemates. The optically active forms can be obtained for example by resolution of the racemates, by asymmetric synthesis or asymmetric chromatography (chromatography with a chiral adsorbent or eluant). That is, certain of the disclosed compounds may exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement. Enantiomers are pairs of stereoisomers whose mirror images are not superimposable, most commonly because they contain an asymmetrically substituted carbon atom that acts as a chiral center. "Enantiomer" means one of a pair of molecules that are mirror images of each other and are not superimposable. Diastereomers are stereoisomers that are not related as mirror images, most commonly because they contain two or more asymmetrically substituted carbon atoms and represent the configuration of substituents around one or more chiral carbon atoms. Enantiomers of a compound can be prepared, for example, by separating an enantiomer from a racemate using one or more well-known techniques and methods, such as, for example, chiral chromatography and separation methods based thereon. The appropriate technique and/or method for separating an enantiomer of a compound described herein from a racemic mixture can be readily determined by those of skill in the art. "Racemate" or "racemic mixture" means a compound containing two enantiomers, wherein such mixtures exhibit no optical activity; i.e., they do not rotate the plane of polarized light. “Geometric isomer" means isomers that differ in the orientation of substituent atoms in relationship to a carbon-carbon double bond, to a cycloalkyl ring, or to a bridged bicyclic system. Atoms (other than H) on each side of a carbon- carbon double bond may be in an E (substituents are on opposite sides of the carbon- carbon double bond) or Z (substituents are oriented on the same side) configuration. "R," "S," "S*," "R*," "E," "Z," "cis," and "trans," indicate configurations relative to the core molecule. Certain of the disclosed compounds may exist in atropisomeric forms. Atropisomers are stereoisomers resulting from hindered rotation about single bonds where the steric strain barrier to rotation is high enough to allow for the isolation of the conformers. The compounds of the invention may be prepared as individual isomers by either isomer-specific synthesis or resolved from an isomeric mixture. Conventional resolution techniques include forming the salt of a free base of each isomer of an isomeric pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming the salt of the acid form of each isomer of an isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), forming an ester or amide of each of the isomers of an isomeric pair using an optically pure acid, amine or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or resolving an isomeric mixture of either a starting material or a final product using various well known chromatographic methods. When the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99% or 99.9%) by weight relative to the other stereoisomers. When a single enantiomer is named or depicted by structure, the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by weight optically pure. When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by weight pure. Percent optical purity is the ratio of the weight of the enantiomer or over the weight of the enantiomer plus the weight of its optical isomer. Diastereomeric purity by weight is the ratio of the weight of one diastereomer or over the weight of all the diastereomers. When the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by mole fraction pure relative to the other stereoisomers. When a single enantiomer is named or depicted by structure, the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by mole fraction pure. When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by mole fraction pure. Percent purity by mole fraction is the ratio of the moles of the enantiomer or over the moles of the enantiomer plus the moles of its optical isomer. Similarly, percent purity by moles fraction is the ratio of the moles of the diastereomer or over the moles of the diastereomer plus the moles of its isomer. When a disclosed compound is named or depicted by structure without indicating the stereochemistry, and the compound has at least one chiral center, it is to be understood that the name or structure encompasses either enantiomer of the compound free from the corresponding optical isomer, a racemic mixture of the compound or mixtures enriched in one enantiomer relative to its corresponding optical isomer. When a disclosed compound is named or depicted by structure without indicating the stereochemistry and has two or more chiral centers, it is to be understood that the name or structure encompasses a diastereomer free of other diastereomers, a number of diastereomers free from other diastereomeric pairs, mixtures of diastereomers, mixtures of diastereomeric pairs, mixtures of diastereomers in which one diastereomer is enriched relative to the other diastereomer(s) or mixtures of diastereomers in which one or more diastereomer is enriched relative to the other diastereomers. The invention embraces all of these forms. Definitions In this application, unless otherwise clear from context, (i) the term “a” may be understood to mean “at least one”; (ii) the term “or” may be understood to mean “and/or”; (iii) the terms “comprising” and “including” may be understood to encompass itemized components or steps whether presented by themselves or together with one or more additional components or steps; and (iv) the terms “about” and “approximately” may be understood to permit standard variation as would be understood by those of ordinary skill in the art; and (v) where ranges are provided, endpoints are included. As used herein, the term “administration” refers to the administration of a composition (e.g., a compound, a complex or a preparation that includes a compound or complex as described herein) to a subject or system. Administration to an animal subject (e.g., to a human) may be by any appropriate route. For example, in some embodiments, administration may be bronchial (including by bronchial instillation), buccal, enteral, interdermal, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal and vitreal. As used herein, the term “animal” refers to any member of the animal kingdom. In some embodiments, “animal” refers to humans, at any stage of development. In some embodiments, “animal” refers to non-human animals, at any stage of development. In some embodiments, the non-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, cattle, a primate, and/or a pig). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, and/or worms. In some embodiments, an animal may be a transgenic animal, genetically-engineered animal, and/or a clone. As used herein, the terms “approximately” and “about” are each intended to encompass normal statistical variation as would be understood by those of ordinary skill in the art as appropriate to the relevant context. In certain embodiments, the terms “approximately” or “about” each refer to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of a stated value, unless otherwise stated or otherwise evident from the context (e.g., where such number would exceed 100% of a possible value). Two events or entities are “associated” with one another, as that term is used herein, if the presence, level and/or form of one is correlated with that of the other. For example, a particular entity (e.g., polypeptide) is considered to be associated with a particular disease, disorder, or condition, if its presence, level and/or form correlates with incidence of and/or susceptibility of the disease, disorder, or condition (e.g., across a relevant population). As used herein, the terms “benefit” and “response” are used interchangeably in the context of a subject, such as a human subject undergoing therapy for the treatment of a neurological disorder, for example, amyotrophic lateral sclerosis, frontotemporal degeneration (also referred to as frontotemporal lobar degeneration and frontotemporal dementia), Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’s disease, Inclusion body myopathy with early-onset Paget disease and frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease, and hereditary inclusion body myopathy. The terms “benefit” and “response” refer to any clinical improvement in the subject’s condition. Exemplary benefits in the context of a subject undergoing treatment for a neurological disorder using the compositions and methods described herein (e.g., in the context of a human subject undergoing treatment for a neurological disorder described herein, such as amyotrophic lateral sclerosis, with a cytochrome P450 isoform 51A1 (CYP51A1) inhibitor described herein, such as an inhibitory small molecule, antibody, antigen-binding fragment thereof, or interfering RNA molecule) include the slowing and halting of disease progression, as well as suppression of one or more symptoms associated with the disease. Particularly, in the context of a patient (e.g., a human patient) undergoing treatment for amyotrophic lateral sclerosis with a CYP51A1 inhibitor described herein, examples of clinical “benefits” and “responses” are (i) an improvement in the subject’s condition as assessed using the amyotrophic lateral sclerosis functional rating scale (ALSFRS) or the revised ALSFRS (ALSFRS-R) following administration of the CYP51A1 inhibitor, such as an improvement in the subject’s ALSFRS or ALSFRS-R score within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an improvement in the subject’s ALSFRS or ALSFRS-R score within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the subject, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the subject); (ii) an increase in the subject’s slow vital capacity following administration of the CYP51A1 inhibitor, such as an increase in the subject’s slow vital capacity within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an increase in the subject’s slow vital capacity within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the subject, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the subject); (iii) a reduction in decremental responses exhibited by the subject upon repetitive nerve stimulation, such as a reduction that is observed within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., a reduction that is observed within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the subject, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the subject); (iv) an improvement in the subject’s muscle strength, as assessed, for example, by way of the Medical Research Council muscle testing scale (as described, e.g., in Jagtap et al., Ann. Indian. Acad. Neurol. 17:336-339 (2014), the disclosure of which is incorporated herein by reference as it pertains to measuring patient response to neurological disease treatment), such as an improvement that is observed within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an improvement that is observed within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the subject, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the subject); (v) an improvement in the subject’s quality of life, as assessed, for example, using the amyotrophic lateral sclerosis-specific quality of life (ALS-specific QOL) questionnaire, such as an improvement in the subject’s quality of life that is observed within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an improvement in the subject’s quality of life that is observed within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the subject, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the subject); and (vi) a decrease in the frequency and/or severity of muscle cramps exhibited by the subject, such as a decrease in cramp frequency and/or severity within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., a decrease in cramp frequency and/or severity within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the subject, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the subject). In the practice of the methods of the present invention, an “effective amount” of any one of the compounds of the invention or a combination of any of the compounds of the invention or a pharmaceutically acceptable salt thereof, is administered via any of the usual and acceptable methods known in the art, either singly or in combination. As used herein, the terms “cytochrome P450 isoform 51A1,” “CYP51A1,” and “lanosterol 14- alpha demethylase” are used interchangeably and refer to the enzyme that catalyzes the conversion of lanosterol to 4,4-dimethylcholesta-8(9),14,24-trien-3β-ol, for example, in human subjects. The terms “cytochrome P450 isoform 51A1,” “CYP51A1,” and “lanosterol 14-alpha demethylase” refer not only to wild-type forms of CYP51A1, but also to variants of wild-type CYP51A1 proteins and nucleic acids encoding the same. The amino acid sequence and corresponding mRNA sequence of a wild-type form of human CYP51A1 are provided herein as SEQ ID NOs: 1 and 2, which correspond to GenBank Accession No. AAC50951.1 and NCBI Reference Sequence NO. NM_000786.3, respectively. These sequences are shown in Table 2, below. Table 2. Amino acid and nucleic acid sequences of wild-type human CYP5A1
Figure imgf000037_0001
Figure imgf000038_0001
The terms “cytochrome P450 isoform 51A1,” “CYP51A1,” and “lanosterol 14-alpha demethylase” as used herein include, for example, forms of the human CYP51A1 protein that have an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 1 (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9%, or 100% identical to the amino acid sequence of SEQ ID NO: 1) and/or forms of the human CYP51A1 protein that contain one or more substitutions, insertions, and/or deletions (e.g., one or more conservative and/or nonconservative amino acid substitutions, such as up to 5, 10, 15, 20, 25, or more, conservative or nonconservative amino acid substitutions) relative to a wild-type CYP51A1 protein. Similarly, the terms “cytochrome P450 isoform 51A1,” “CYP51A1,” and “lanosterol 14-alpha demethylase” as used herein include, for example, forms of the human CYP51A1 gene that encode an mRNA transcript having a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 2 (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9%, or 100% identical to the amino acid sequence of SEQ ID NO: 2). As used herein, the terms “cytochrome P450 isoform 51A1 inhibitor,” “CYP51A1 inhibitor,” and “lanosterol 14-alpha demethylase inhibitor” are used interchangeably and refer to substances, such as compounds of Formula I. Inhibitors of this type may, for example, competitively inhibit CYP51A1 activity by specifically binding the CYP51A1 enzyme (e.g., by virtue of the affinity of the inhibitor for the CYP51A1 active site), thereby precluding, hindering, or halting the entry of one or more endogenous substrates of CYP51A1 into the enzyme’s active site. Additional examples of CYP51A1 inhibitors that suppress the activity of the CYP51A1 enzyme include substances that may bind CYP51A1 at a site distal from the active site and attenuate the binding of endogenous substrates to the CYP51A1 active site by way of a change in the enzyme’s spatial conformation upon binding of the inhibitor. In addition to encompassing substances that modulate CYP51A1 activity, the terms “cytochrome P450 isoform 51A1 inhibitor,” “CYP51A1 inhibitor,” and “lanosterol 14-alpha demethylase inhibitor” refer to substances that reduce the concentration and/or stability of CYP51A1 mRNA transcripts in vivo, as well as those that suppress the translation of functional CYP51A1 enzyme. As used herein, the term “CYP51A1-associated disorder” refers to an undesired physiological condition, disorder, or disease that is associated with and/or mediated at least in part by CYP51A1. In some instances, CYP51A1-associated disorders are associated with excess CYP51A1 levels and/or activity. Exemplary CYP51A1-associated disorders include CYP51A1-associated disorders include but are not limited to central nervous system (CNS) disorders, dementia, Alzheimer's Disease, chronic traumatic encephalopathy, FTLD-TDP, LATE, or frontotemporal lobar degeneration. As used herein, the term “combination therapy” refers to those situations in which a subject is simultaneously exposed to two or more therapeutic agents. In some embodiments, two or more compounds may be administered simultaneously; in some embodiments, such compounds may be administered sequentially; in some embodiments, such compounds are administered in overlapping dosing regimens. As used herein, the term “dosage form” refers to a physically discrete unit of an active compound (e.g., a therapeutic or diagnostic agent) for administration to a subject. Each unit contains a predetermined quantity of active agent. In some embodiments, such quantity is a unit dosage amount (or a whole fraction thereof) appropriate for administration in accordance with a dosing regimen that has been determined to correlate with a desired or beneficial outcome when administered to a relevant population (i.e., with a therapeutic dosing regimen). Those of ordinary skill in the art appreciate that the total amount of a therapeutic composition or compound administered to a particular subject is determined by one or more attending physicians and may involve administration of multiple dosage forms. As used herein, the term “dosing regimen” refers to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time. In some embodiments, a given therapeutic compound has a recommended dosing regimen, which may involve one or more doses. In some embodiments, a dosing regimen comprises a plurality of doses each of which are separated from one another by a time period of the same length; in some embodiments, a dosing regimen comprises a plurality of doses and at least two different time periods separating individual doses. In some embodiments, all doses within a dosing regimen are of the same unit dose amount. In some embodiments, different doses within a dosing regimen are of different amounts. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount different from the first dose amount. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount same as the first dose amount. In some embodiments, a dosing regimen is correlated with a desired or beneficial outcome when administered across a relevant population (i.e., is a therapeutic dosing regimen). As used herein, the term “neuromuscular disorder” refers to a disease impairing the ability of one or more neurons to control the activity of an associated muscle. Examples of neuromuscular disorders are amyotrophic lateral sclerosis, congenital myasthenic syndrome, congenital myopathy, cramp fasciculation syndrome, Duchenne muscular dystrophy, glycogen storage disease type II, hereditary spastic paraplegia, inclusion body myositis, Isaac's Syndrome, Kearns-Sayre syndrome, Lambert–Eaton myasthenic syndrome, mitochondrial myopathy, muscular dystrophy, myasthenia gravis, myotonic dystrophy, peripheral neuropathy, spinal and bulbar muscular atrophy, spinal muscular atrophy, Stiff person syndrome, Troyer syndrome, and Guillain–Barré syndrome, among others. The term “pharmaceutical composition,” as used herein, represents a composition containing a compound described herein formulated with a pharmaceutically acceptable excipient, and manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for the treatment of disease in a mammal. Pharmaceutical compositions can be formulated, for example, for oral administration in unit dosage form (e.g., a tablet, capsule, caplet, gelcap, or syrup); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate emboli and in a solvent system suitable for intravenous use); or in any other pharmaceutically acceptable formulation. A “pharmaceutically acceptable excipient,” as used herein, refers any ingredient other than the compounds described herein (for example, a vehicle capable of suspending or dissolving the active compound) and having the properties of being substantially nontoxic and non-inflammatory in a patient. Excipients may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspensing or dispersing agents, sweeteners, and waters of hydration. As used herein, the term “pharmaceutically acceptable salt” means any pharmaceutically acceptable salt of the compound of formula (I). For example, pharmaceutically acceptable salts of any of the compounds described herein include those that are within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and in Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. The salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting a free base group with a suitable organic acid. The compounds of the invention may have ionizable groups so as to be capable of preparation as pharmaceutically acceptable salts. These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds of the invention be prepared from inorganic or organic bases. Frequently, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases and methods for preparation of the appropriate salts are well-known in the art. Salts may be prepared from pharmaceutically acceptable non-toxic acids and bases including inorganic and organic acids and bases. The term “pure” means substantially pure or free of unwanted components (e.g., other compounds and/or other components of a cell lysate), material defilement, admixture or imperfection. A variety of clinical indicators can be used to identify a patient as “at risk” of developing a particular neurological disease. Examples of patients (e.g., human patients) that are “at risk” of developing a neurological disease, such as amyotrophic lateral sclerosis, frontotemporal degeneration, Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’s disease, Inclusion body myopathy with early-onset Paget disease and frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease, and hereditary inclusion body myopathy, include (i) subjects exhibiting or prone to exhibit aggregation of TAR-DNA binding protein (TDP)-43, and (ii) subjects expressing a mutant form of TDP-43 containing a mutation associated with TDP-43 aggregation and toxicity, such as a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D. Subjects that are “at risk” of developing amyotrophic lateral sclerosis may exhibit one or both of these characteristics, for example, prior to the first administration of a CYP51A1 inhibitor in accordance with the compositions and methods described herein. As used herein, the terms “TAR-DNA binding protein-43” and “TDP-43” are used interchangeably and refer to the transcription repressor protein involved in modulating HIV-1 transcription and alternative splicing of the cystic fibrosis transmembrane conductance regulator (CFTR) pre-mRNA transcript, for example, in human subjects. The terms “TAR-DNA binding protein-43” and “TDP-43” refer not only to wild-type forms of TDP-43, but also to variants of wild-type TDP-43 proteins and nucleic acids encoding the same. The amino acid sequence and corresponding mRNA sequence of a wild-type form of human TDP-43 are provided herein as SEQ ID NOs: 3 and 4, which correspond to NCBI Reference Sequence NOs. NM_007375.3 and NP_031401.1, respectively. These sequences are shown in Table 3, below. Table 3. Amino acid and nucleic acid sequences of wild-type human TDP-43
Figure imgf000041_0001
Figure imgf000042_0001
The terms “TAR-DNA binding protein-43” and “TDP-43” as used herein include, for example, forms of the human TDP-43 protein that have an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 3 (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9%, or 100% identical to the amino acid sequence of SEQ ID NO: 3) and/or forms of the human TDP-43 protein that contain one or more substitutions, insertions, and/or deletions (e.g., one or more conservative and/or nonconservative amino acid substitutions, such as up to 5, 10, 15, 20, 25, or more, conservative or nonconservative amino acid substitutions) relative to a wild- type TDP-43 protein. For instance, patients that may be treated for a neurological disorder as described herein, such as amyotrophic lateral sclerosis, frontotemporal degeneration, Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’s disease, Inclusion body myopathy with early-onset Paget disease and frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease, and hereditary inclusion body myopathy, include human patients that express a form of TDP-43 having a mutation associated with elevated TDP-43 aggregation and toxicity, such as a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D. Similarly, the terms “TAR-DNA binding protein-43” and “TDP-43” as used herein include, for example, forms of the human TDP-43 gene that encode an mRNA transcript having a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 4 (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9%, or 100% identical to the amino acid sequence of SEQ ID NO: 4). As used herein, the term “subject” refers to any organism to which a composition in accordance with the invention may be administered, e.g., for experimental, diagnostic, prophylactic, and/or therapeutic purposes. Typical subjects include any animal (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans). A subject may seek or be in need of treatment, require treatment, be receiving treatment, be receiving treatment in the future, or be a human or animal who is under care by a trained professional for a particular disease or condition. As used herein, the terms "treat," "treated," or "treating" mean both therapeutic treatment and prophylactic or preventative measures wherein the object is to prevent or slow down (lessen) an undesired physiological condition, disorder, or disease, or obtain beneficial or desired clinical results. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of the extent of a condition, disorder, or disease; stabilized (i.e., not worsening) state of condition, disorder, or disease; delay in onset or slowing of condition, disorder, or disease progression; amelioration of the condition, disorder, or disease state or remission (whether partial or total), whether detectable or undetectable; an amelioration of at least one measurable physical parameter, not necessarily discernible by the patient; or enhancement or improvement of condition, disorder, or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival as compared to expected survival if not receiving treatment. A “therapeutic regimen” refers to a dosing regimen whose administration across a relevant population is correlated with a desired or beneficial therapeutic outcome. The term “therapeutically effective amount” means an amount that is sufficient, when administered to a population suffering from or susceptible to a disease, disorder, and/or condition in accordance with a therapeutic dosing regimen, to treat the disease, disorder, and/or condition. In some embodiments, a therapeutically effective amount is one that reduces the incidence and/or severity of, and/or delays onset of, one or more symptoms of the disease, disorder, and/or condition. Those of ordinary skill in the art will appreciate that the term “therapeutically effective amount” does not in fact require successful treatment be achieved in a particular individual. Rather, a therapeutically effective amount may be that amount that provides a particular desired pharmacological response in a significant number of subjects when administered to patients in need of such treatment. It is specifically understood that particular subjects may, in fact, be “refractory” to a “therapeutically effective amount.” To give but one example, a refractory subject may have a low bioavailability such that clinical efficacy is not obtainable. In some embodiments, reference to a therapeutically effective amount may be a reference to an amount as measured in one or more specific tissues (e.g., a tissue affected by the disease, disorder or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine, etc). Those of ordinary skill in the art will appreciate that, in some embodiments, a therapeutically effective amount may be formulated and/or administered in a single dose. In some embodiments, a therapeutically effective amount may be formulated and/or administered in a plurality of doses, for example, as part of a dosing regimen. Brief Description of the Drawings FIGS.1A – 1C demonstrate that the viability of a yeast TDP-43 model is restored by the Erg11 inhibitor, fluconazole. (FIG.1A) Structure of the Erg11 inhibitor and anti-fungal, fluconazole. (FIG.1B) Fluconazole rescues viability of TDP-43-expressing yeast using a resazurin-reduction endpoint. A 2-fold serial dilution of fluconazole was applied to TDP-43-expressing yeast for 24 hours prior to analysis. (FIG. 1C) Wild-type yeast cultures were treated with fluconazole for eight hours prior to HPLC analysis for lanosterol and ergosterol. Data are expressed as the area under the curve (AUC) normalized to cell mass based on optical density of cultures at 600 nm. Fluconazole treatment reduces ergosterol, while simultaneously leading to an increase in the Erg11 substrate, lanosterol. FIG.2 shows the structures of compounds used in primary rat cortical neuron TDP-43 wild type and Q331K mutant survival studies. FIGS.3A and 3B demonstrate that compound A promotes survival in primary rat cortical neurons transfected with wild-type TDP-43. Rat primary cortical neurons were co-transfected with a red fluorescent protein (RFP) as a morphological marker and either control (empty vector) or wild-type TDP- 43 expression plasmids and treated with vehicle (DMSO) or a titration of compound A. (FIG.3A) Risk of neuron death plots. The lifetime of each neuron was determined by either loss of RFP signal or morphological indicators of death such as loss of neurites and cell blebbing and used to generate cumulative hazard plots of risk of death over time (hrs) post-transfection. (FIG.3B) Forest plots. Hazard ratios for each treatment group (relative to TDP-43 DMSO group) were determined by cox regression analysis and used to generate forest plots. Hazard ratios (HR) < 1 in which the confidence interval (CI) does not encompass 1 represent treatments that significantly reduce probability of neuron death relative to the TDP-43 DMSO control. P, p-value. FIGS.4A and 4B demonstrate that compound A promotes survival in primary rat cortical neurons transfected with Q331K Mutant TDP-43. Rat primary cortical neurons were co-transfected with a red fluorescent protein (RFP) as a morphological marker and either control (empty vector) or Q331K mutant TDP-43 expression plasmids and treated with vehicle (DMSO) or a titration of compound A. (FIG.4A) Risk of neuron death plots. The lifetime of each neuron was determined by either loss of RFP signal or morphological indicators of death such as loss of neurites and cell blebbing and used to generate cumulative hazard plots of risk of death over time (hrs) post-transfection. (FIG.4B) Forest plots. Hazard ratios for each treatment group (relative to TDP-43 DMSO group) were determined by cox regression analysis and used to generate forest plots. Hazard ratios (HR) < 1 in which the confidence interval (CI) does not encompass 1 represent treatments that significantly reduce probability of neuron death relative to the TDP-43 DMSO control. P, p-value. FIGS.5A and 5B demonstrate that compound B promotes survival in primary rat cortical neurons transfected with wild-type TDP-43. Rat primary cortical neurons were co-transfected with a red fluorescent protein (RFP) as a morphological marker and either control (empty vector) or wild type TDP- 43 expression plasmids and treated with vehicle (DMSO) or a titration of compound B. (FIG.5A) Risk of neuron death plots. The lifetime of each neuron was determined by either loss of RFP signal or morphological indicators of death such as loss of neurites and cell blebbing and used to generate cumulative hazard plots of risk of death over time (hrs) post-transfection. (FIG.5B) Forest plots. Hazard ratios for each treatment group (relative to TDP-43 DMSO group) were determined by cox regression analysis and used to generate forest plots. Hazard ratios (HR) < 1 in which the confidence interval (CI) does not encompass 1 represent treatments that significantly reduce probability of neuron death relative to the TDP-43 DMSO control. P, p-value. Detailed Description The present invention features compositions and methods for treating neurological disorders, such as amyotrophic lateral sclerosis and other neuromuscular disorders, as well as frontotemporal degeneration, Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’s disease, Inclusion body myopathy with early-onset Paget disease and frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease, and hereditary inclusion body myopathy among others. Particularly, the invention provides inhibitors of cytochrome P450 isoform 51A1 (CYP51A1), also referred to herein as lanosterol 14-alpha demethylase, that may be administered to a patient (e.g., a human patient) so as to treat or prevent a neurological disorder, such as one or more of the foregoing conditions. In the context of therapeutic treatment, the CYP51A1 inhibitor may be administered to the patient to alleviate one or more symptoms of the disorder and/or to remedy an underlying molecular pathology associated with the disease, such as to suppress or prevent aggregation of TAR-DNA binding protein (TDP)-43. The disclosure herein is based, in part, on the discovery that CYP51A1 inhibition modulates TDP- 43 aggregation in vivo. Suppression of TDP-43 aggregation exerts beneficial effects in patients suffering from a neurological disorder. Many pathological conditions have been correlated with TDP-43-promoted aggregation and toxicity, such as amyotrophic lateral sclerosis, frontotemporal degeneration, Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’s disease, IBMPFD, sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease, and hereditary inclusion body myopathy. Without being limited by mechanism, by administering an inhibitor of CYP51A1, patients suffering from diseases associated with TDP-43 aggregation and toxicity may be treated, for example, due to the suppression of TDP-43 aggregation induced by the CYP51A1 inhibitor. Patients that are likely to respond to CYP51A1 inhibition as described herein include those that have or are at risk of developing TDP-43 aggregation, such as those that express a mutant form of TDP- 43 associated with TDP-43 aggregation and toxicity in vivo. Examples of such mutations in TDP-43 that have been correlated with elevated TDP-43 aggregation and toxicity include Q331K, M337V, Q343R, N345K, R361S, and N390D, among others. The compositions and methods described herein thus provide the additional clinical benefit of enabling the identification of patients that are likely to respond to CYP51A1 inhibitor therapy, as well as processes for treating these patients accordingly. The sections that follow provide a description of exemplary CYP51A1 inhibitors that may be used in conjunction with the compositions and methods disclosed herein. The sections below additionally provide a description of various exemplary routes of administration and pharmaceutical compositions that may be used for delivery of these substances for the treatment of a neurological disorder. CYP51A1 Inhibitors Exemplary CYP51A1 inhibitors described herein include compounds having a structure according to Formula I:
Figure imgf000045_0001
Formula I wherein n is 0, 1, 2, 3, or 4; X is S or O; each R1 is, independently, halo, cyano, hydroxy, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, -SO2-optionally substituted C1-C6 alkyl, or -CO2-optionally substituted C1-C6 alkyl; R2 is optionally substituted C2-C9 heteroaryl; and R3 is optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C9 heterocyclyl, optionally substituted C1-C6 alkyl C3-C8 cycloalkyl, optionally substituted C1-C6 heteroalkyl C3-C8 cycloalkyl, optionally substituted C1-C6 alkyl C2-C9 heteroaryl, optionally substituted C1-C6 heteroalkyl C2-C9 heteroaryl, optionally substituted C1-C6 alkyl C2-C9 heterocyclyl, optionally substituted C1-C6 heteroalkyl C2-C9 heterocyclyl, optionally substituted C1-C6 alkyl C6-C10 aryl, or optionally substituted C1-C6 heteroalkyl C6-C10 aryl, or pharmaceutically acceptable salts thereof. In some embodiments, the compound has the structure of any one of compounds 1-273 in Table 1. Other embodiments, as well as exemplary methods for the synthesis or production of these compounds, are described herein. Methods of Treatment Suppression of CYP51A1 Activity and TDP-43 Aggregation to Treat Neurological Disorders Using the compositions and methods described herein, a patient suffering from a neurological disorder may be administered a CYP51A1 inhibitor, such as a small molecule, antibody, antigen-binding fragment thereof, or interfering RNA molecule described herein, so as to treat the disorder and/or to suppress one or more symptoms associated with the disorder. Exemplary neurological disorders that may be treated using the compositions and methods described herein are, without limitation, amyotrophic lateral sclerosis, frontotemporal degeneration, Alzheimer’s disease, Parkinson’s disease, dementia with Lewy Bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington’s disease, IBMPFD, sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease, and hereditary inclusion body myopathy, as well as neuromuscular diseases such as congenital myasthenic syndrome, congenital myopathy, cramp fasciculation syndrome, Duchenne muscular dystrophy, glycogen storage disease type II, hereditary spastic paraplegia, inclusion body myositis, Isaac's Syndrome, Kearns-Sayre syndrome, Lambert–Eaton myasthenic syndrome, mitochondrial myopathy, muscular dystrophy, myasthenia gravis, myotonic dystrophy, peripheral neuropathy, spinal and bulbar muscular atrophy, spinal muscular atrophy, Stiff person syndrome, Troyer syndrome, and Guillain–Barré syndrome. The present disclosure is based, in part, on the discovery that CYP51A1 inhibitors, such as the agents described herein, are capable of attenuating TDP-43 aggregation in vivo. TDP-43-promoted aggregation and toxicity have been associated with various neurological diseases. The discovery that CYP51A1 inhibitors modulate TDP-43 aggregation provides an important therapeutic benefit. Using a CYP51A1 inhibitor, such as a CYP51A1 inhibitor described herein, a patient suffering from a neurological disorder or at risk of developing such a condition may be treated in a manner that remedies an underlying molecular etiology of the disease. Without being limited by mechanism, the compositions and methods described herein can be used to treat or prevent such neurological conditions, for example, by suppressing the TDP-43 aggregation that promotes pathology. Additionally, the compositions and methods described herein provide the beneficial feature of enabling the identification and treatment of patients that are likely to respond to CYP51A1 inhibitor therapy. For example, in some embodiments, a patient (e.g., a human patient suffering from or at risk of developing a neurological disease described herein, such as amyotrophic lateral sclerosis) is administered a CYP51A1 inhibitor if the patient is identified as likely to respond to this form of treatment. Patients may be identified as such on the basis, for example, of susceptibility to TDP-43 aggregation. In some embodiments, the patient is identified is likely to respond to CYP51A1 inhibitor treatment based on the isoform of TDP-43 expressed by the patient. For example, patients expressing TDP-43 isoforms having a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D, among others, are more likely to develop TDP-43-promoted aggregation and toxicity relative to patients that do not express such isoforms of TDP-43. Using the compositions and methods described herein, a patient may be identified as likely to respond to CYP51A1 inhibitor therapy on the basis of expressing such an isoform of TDP-43, and may subsequently be administered a CYP51A1 inhibitor so as to treat or prevent one or more neurological disorders, such as one or more of the neurological disorders described herein. Assessing Patient Response A variety of methods known in the art and described herein can be used to determine whether a patient having a neurological disorder (e.g., a patient at risk of developing TDP-43 aggregation, such as a patient expressing a mutant form of TDP-43 having a mutation associated with elevated TDP-43 aggregation and toxicity, for example, a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D) is responding favorably to CYP51A1 inhibition. For example, successful treatment of a patient having a neurological disease, such as amyotrophic lateral sclerosis, with a CYP51A1 inhibitor described herein may be signaled by: (i) an improvement in condition as assessed using the amyotrophic lateral sclerosis functional rating scale (ALSFRS) or the revised ALSFRS (ALSFRS-R), such as an improvement in the patient’s ALSFRS or ALSFRS-R score within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an improvement in the patient’s ALSFRS or ALSFRS-R score within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the patient); (ii) an increase in slow vital capacity, such as an increase in the patient’s slow vital capacity within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an increase in the patient’s slow vital capacity within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the patient); (iii) a reduction in decremental responses exhibited by the patient upon repetitive nerve stimulation, such as a reduction that is observed within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., a reduction that is observed within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the patient); (iv) an improvement in muscle strength, as assessed, for example, by way of the Medical Research Council muscle testing scale (as described, e.g., in Jagtap et al., Ann. Indian. Acad. Neurol. 17:336-339 (2014), the disclosure of which is incorporated herein by reference as it pertains to measuring patient response to neurological disease treatment), such as an improvement that is observed within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an improvement that is observed within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the patient); (v) an improvement in quality of life, as assessed, for example, using the amyotrophic lateral sclerosis-specific quality of life (ALS-specific QOL) questionnaire, such as an improvement in the patient’s quality of life that is observed within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., an improvement in the subject’s quality of life that is observed within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the patient); (vi) a decrease in the frequency and/or severity of muscle cramps, such as a decrease in cramp frequency and/or severity within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., a decrease in cramp frequency and/or severity within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the patient); and/or (vii) a decrease in TDP-43 aggregation, such as a decrease in TDP-43 aggregation within one or more days, weeks, or months following administration of the CYP51A1 inhibitor (e.g., a decrease in TDP- 43 aggregation within from about 1 day to about 48 weeks (e.g., within from about 2 days to about 36 weeks, from about 4 weeks to about 24 weeks, from about 8 weeks to about 20 weeks, or from about 12 weeks to about 16 weeks), or more, following the initial administration of the CYP51A1 inhibitor to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, or more, following the initial administration of the CYP51A1 inhibitor to the patient. Combination Formulations and Uses Thereof The compounds of the invention can be combined with one or more therapeutic agents. In particular, the therapeutic agent can be one that treats or prophylactically treats any neurological disorder described herein. Combination Therapies A compound of the invention can be used alone or in combination with other agents that treat neurological disorders or symptoms associated therewith, or in combination with other types of treatment to treat, prevent, and/or reduce the risk of any neurological disorders. In combination treatments, the dosages of one or more of the therapeutic compounds may be reduced from standard dosages when administered alone. For example, doses may be determined empirically from drug combinations and permutations or may be deduced by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6, 2005). In this case, dosages of the compounds when combined should provide a therapeutic effect. Pharmaceutical Compositions The compounds of the invention are preferably formulated into pharmaceutical compositions for administration to human subjects in a biologically compatible form suitable for administration in vivo. Accordingly, in another aspect, the present invention provides a pharmaceutical composition comprising a compound of the invention in admixture with a suitable diluent, carrier, or excipient. The compounds of the invention may be used in the form of the free base, in the form of salts, solvates, and as prodrugs. All forms are within the scope of the invention. In accordance with the methods of the invention, the described compounds or salts, solvates, or prodrugs thereof may be administered to a patient in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art. The compounds of the invention may be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump, or transdermal administration and the pharmaceutical compositions formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal, and topical modes of administration. Parenteral administration may be by continuous infusion over a selected period of time. A compound of the invention may be orally administered, for example, with an inert diluent or with an assimilable edible carrier, or it may be enclosed in hard or soft shell gelatin capsules, or it may be compressed into tablets, or it may be incorporated directly with the food of the diet. For oral therapeutic administration, a compound of the invention may be incorporated with an excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, and wafers. A compound of the invention may also be administered parenterally. Solutions of a compound of the invention can be prepared in water suitably mixed with a surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, DMSO and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms. Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington’s Pharmaceutical Sciences (2003, 20th ed.) and in The United States Pharmacopeia: The National Formulary (USP 24 NF19), published in 1999. The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form must be sterile and must be fluid to the extent that may be easily administered via syringe. Compositions for nasal administration may conveniently be formulated as aerosols, drops, gels, and powders. Aerosol formulations typically include a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually presented in single or multidose quantities in sterile form in a sealed container, which can take the form of a cartridge or refill for use with an atomizing device. Alternatively, the sealed container may be a unitary dispensing device, such as a single dose nasal inhaler or an aerosol dispenser fitted with a metering valve which is intended for disposal after use. Where the dosage form comprises an aerosol dispenser, it will contain a propellant, which can be a compressed gas, such as compressed air or an organic propellant, such as fluorochlorohydrocarbon. The aerosol dosage forms can also take the form of a pump-atomizer. Compositions suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, where the active ingredient is formulated with a carrier, such as sugar, acacia, tragacanth, gelatin, and glycerine. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base, such as cocoa butter. The compounds of the invention may be administered to an animal, e.g., a human, alone or in combination with pharmaceutically acceptable carriers, as noted herein, the proportion of which is determined by the solubility and chemical nature of the compound, chosen route of administration, and standard pharmaceutical practice. Dosages The dosage of the compounds of the invention, and/or compositions comprising a compound of the invention, can vary depending on many factors, such as the pharmacodynamic properties of the compound; the mode of administration; the age, health, and weight of the recipient; the nature and extent of the symptoms; the frequency of the treatment, and the type of concurrent treatment, if any; and the clearance rate of the compound in the animal to be treated. One of skill in the art can determine the appropriate dosage based on the above factors. The compounds of the invention may be administered initially in a suitable dosage that may be adjusted as required, depending on the clinical response. In general, satisfactory results may be obtained when the compounds of the invention are administered to a human at a daily dosage of, for example, between 0.05 mg and 3000 mg (measured as the solid form). Dose ranges include, for example, between 10-1000 mg. Alternatively, the dosage amount can be calculated using the body weight of the patient. For example, the dose of a compound, or pharmaceutical composition thereof, administered to a patient may range from 0.1-50 mg/kg. EXAMPLES List of Abbreviations: • NaBH4 = sodium borohydride • EtOH = ethanol • MeOH = methanol • NEt3 = triethylamine • DCM = dichloromethane • NaHB(OAc)3 = sodium triacetoxyborohydride • AcOH = acetic acid • HCl = hydrochloric acid • DMF = N,N-dimethylformamide • THF = tetrahydrofuran • T3P = propylphosphonic anhydride • DIPEA = N,N-diisopropylethylamine • Pyr = pyridine • TFA = trifluoroacetic acid • Ti(O-iPr)4 = titanium(IV) isopropoxide • HATU = N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide • HBTU = N,N,N',N'-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate • DMAP = 4-(dimethylamino)pyridine • KSCN = potassium thiocyanate • Br2 = bromine • NaCNBH3 = sodium cyanoboroydride • NaH = sodium hydride • Boc2O = di(tert-butyl) carbonate • HOBT = 1-hydroxybenzotriazole • DCC = 1,3-dicyclohexylcarbodiimide • EDC = 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride • LiOH x H2O = lithium hydroxide hydrate General Schemes: General Scheme 1
Figure imgf000052_0001
Condensation of appropriately substituted amine I with appropriately substituted aldehyde II in the presence of a variety of reducing agents (e.g. sodium borohydride) gives appropriately substituted amine III. Reaction of amine III with appropriately substituted acid IV under a variety of coupling conditions (e.g. HATU), or acyl chloride IV generated in situ (e.g. with thionyl chloride), or pre-generated acyl chloride IV under basic conditions (e.g. triethylamine) yields amide V. R1, R2, or R3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions. General Scheme 2
Figure imgf000052_0002
An appropriately substituted amine I and alkyl halide II can be reacted under basic conditions (e.g. potassium carbonate) to give appropriately substituted amine III. Reaction of amine III with appropriately substituted acyl chloride IV under basic conditions (e.g. triethylamine) affords amide V. R1, R2, or R3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions. General Scheme 3
Figure imgf000053_0001
Reaction of appropriately substituted amine I with appropriately substituted acid II under a variety of coupling conditions (e.g. HBTU) yields amide III. Reaction of amide III with appropriately substituted alkyl halide IV under basic conditions (e.g. potassium carbonate) affords desired amide V. R1, R2, or R3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions. General Scheme 4
Figure imgf000053_0002
An appropriately substituted amine I is reacted with potassium thiocyanate II to give appropriately substituted benzothiazol-2-amine III. Condensation of benzothiazol-2-amine III with appropriately substituted aldehyde IV in the presence of a variety of reducing agents (e.g. sodium borohydride) gives appropriately substituted amine V. Reaction of amine V with appropriately substituted acyl chloride VI under basic conditions (e.g. triethylamine) affords amide VII. R1, R2, or R3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions. General Scheme 5
Figure imgf000053_0003
Condensation of appropriately substituted amine I with appropriately substituted aldehyde II in the presence of a variety of reducing agents (e.g. sodium borohydride) gives appropriately substituted amine III. R1, R2, or R3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions. General Scheme 6
Figure imgf000054_0001
An appropriately substituted aryl halide I and appropriately substituted amine II can be reacted under basic (e.g. potassium carbonate) and/or microwave conditions to give appropriately substituted amine III. Reaction of amine III with appropriately substituted acyl chloride IV under basic conditions (e.g. triethylamine) affords amide V. R1, R2, or R3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions. General Scheme 7
Figure imgf000054_0002
Condensation of appropriately substituted amine I with appropriately substituted aldehyde II in the presence of a variety of reducing agents (e.g. sodium borohydride) gives appropriately substituted amine III. Coupling of amine III and appropriately substituted amine IV with triphosgene under basic conditions (e.g. triethylamine) yields urea V. R1, R2, or R3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions.
Figure imgf000054_0003
An appropriately substituted amine I is reacted with appropriately substituted acid II under a variety of coupling conditions (e.g. HATU), or acyl chloride II under basic conditions (e.g. triethylamine), to yield amide or carbamate III. R1, R2, or R3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions. General Scheme 9
Figure imgf000055_0004
Piperidine-4-carboxylic acid I is reacted with appropriately substituted acyl chloride II under basic conditions (e.g. triethylamine) to yield appropriately substituted carboxylic acid III. Carboxylic acid III is coupled with appropriately substituted amine IV under a variety of coupling conditions (e.g. HATU) to yield amide or carbamate V. R1, R2, or R3 may contain N-protecting groups (e.g. trityl) that require removal under acidic conditions. General Scheme 10
Figure imgf000055_0001
An appropriately substituted amine I can be reacted with appropriately substituted alcohol II and triphosgene under basic conditions (e.g. triethylamine) to yield carbamate III. R1, R2, or R3 may contain N- protecting groups (e.g. trityl) that require removal under acidic conditions. Example 1. Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
Figure imgf000055_0002
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000055_0003
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified by flash column chromatography through silica gel using a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N- (pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000056_0001
To a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (1.34 g, 5.0 mmol), 4- cyanobenzoic acid (0.740 g, 5.0 mmol), N-N,diisopropylethylamine (1.50 g, 15 mmol) in tetrahydrofuran (50 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (2.80 g, 7.5 mmol). The mixture was heated to 90 °C and stirred for 4 hours. Then, the solvent was removed under the reduced pressure and the residue was purified by flash column chromatography through silica gel with a gradient of 0-50% ethyl acetate in petroleum ether. The elutant was then was recrystallized from ethanol to offer 4-cyano-N-(6- ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (1.60 g, 4.0 mmol, 80 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.43 (dd, J = 5.0, 1.5 Hz, 1H), 8.33 (d, J = 1.7 Hz, 1H), 7.96 (d, J = 8.3 Hz, 2H), 7.87 (s, 1H), 7.76 (d, J = 8.3 Hz, 2H), 7.69 (d, J = 8.3 Hz, 1H), 7.57 (d, J = 8.0 Hz, 1H), 7.39 – 7.17 (m, 2H), 5.40 (s, 2H), 2.72 – 2.68 (m, 2H), 1.23 – 1.15 (m, 3H); LCMS (ESI) m/z: 399.1 [M+H]+. Example 2. Preparation of N-(6-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide
Figure imgf000056_0002
Step 1: Preparation of 6-methyl-N-(3-pyridylmethyl)-1,3-benzothiazol-2-amine
Figure imgf000056_0003
To a stirred solution of 6-methyl-1,3-benzothiazol-2-amine (0.500 g, 3.0 mmol) in N,N- dimethylformamide (10 mL) was added 3-(chloromethyl)pyridine (0.499 g, 3.0 mmol), cesium carbonate (1.98 g, 6.1 mmol). The mixture was stirred at 100 °C for 16 hours. LCMS showed product formation.15 mL of water was added to the reaction, the reaction mixture was extracted with ethyl acetate (2x 30 mL). The combined organic layers were washed with brine (15 mL) and dried over sodium sulfate. The combined organic layers were concentrated to dryness. The crude product was purified by flash column chromatography in a gradient of 0-80% ethyl acetate in petroleum ether through 20 g of silica gel. Product 6-methyl-N-(3-pyridylmethyl)-1,3-benzothiazol-2-amine (0.200 g, 0.78 mmol, 26%) was obtained as a yellow solid.1H NMR (400 MHz, Chloroform-d) δ = 8.66 (d, J = 1.5 Hz, 1H), 8.56 (dd, J = 1.4, 4.7 Hz, 1H), 7.76 (br d, J = 7.7 Hz, 1H), 7.43 (d, J = 8.4 Hz, 1H), 7.40 (s, 1H), 7.31 - 7.28 (m, 1H), 7.12 (dd, J = 1.1, 8.2 Hz, 1H), 5.79 (br s, 1H), 4.69 (s, 2H), 2.40 (s, 3H). Step 2: Preparation of N-(6-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide
Figure imgf000057_0001
To a solution of 6-methyl-N-(3-pyridylmethyl)-1,3-benzothiazol-2-amine (150 mg, 0.59 mmol) in dichloromethane (5 mL) was added triethylamine (238 mg, 2.4 mmol), cyclohexanecarbonyl chloride (86.0 mg, 0.59 mmol) at 0 °C. The mixture was stirred at room temperature for 30 minutes. LCMS showed the reaction was complete. The reaction mixture was concentrated to dryness. Crude product was purified by prep-HPLC (Waters Xbridge 150*255u column; 50-80% acetonitrile in a 10 mM ammonium bicarbonate solution in water, 10 min gradient). Product N-(6-methyl-1,3-benzothiazol-2-yl)-N- (3-pyridylmethyl)cyclohexanecarboxamide (112 mg, 0.31 mmol, 52 %) was obtained as a pale yellow solid.1H NMR (400 MHz, Methanol-d4) δ = 8.54 (d, J=1.6 Hz, 1H), 8.46 (dd, J=1.3, 4.9 Hz, 1H), 7.74 (br d, J=7.9 Hz, 1H), 7.66 (s, 1H), 7.61 (d, J=8.3 Hz, 1H), 7.41 (dd, J=4.9, 7.9 Hz, 1H), 7.24 (dd, J=1.2, 8.3 Hz, 1H), 5.70 (br s, 2H), 2.83 (tt, J=3.2, 11.4 Hz, 1H), 2.45 (s, 3H), 1.82 - 1.65 (m, 5H), 1.62 - 1.46 (m, 2H), 1.35 - 1.17 (m, 3H); LCMS (ESI) m/z: 366.2 [M+H]+. Example 3. Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide
Figure imgf000057_0002
Step 1: Preparation of 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000057_0003
To a solution of nicotinaldehyde (1.19 g, 11 mmol), 6-methoxybenzo[d]thiazol-2-amine (2.00 g, 11 mmol) in toluene (10 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (2.11 g, 56 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol- 2-amine (1.10 g, 4.1 mmol, 36%) as a white solid. LCMS (ESI) m/z: 272.1 [M+H]+. Step 2: Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide
Figure imgf000058_0001
To a solution of 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (150 mg, 0.55 mmol), N,N-diisopropylethylamine (214 mg, 1.7 mmol) in dichloromethane (10 mL) was added cyclohexanecarbonyl chloride (89.0 mg, 0.61 mmol) at room temperature. The mixture was stirred at room temperature for 2 hours, then the solvent was removed under the reduced pressure. The residue was purified by prep-HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(6-methoxybenzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)cyclohexanecarboxamide (39.8 mg, 0.10 mmol, 19 %) as a light gray solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.53 (d, J = 26.3 Hz, 2H), 7.79 – 7.53 (m, 3H), 7.50 – 7.26 (m, 1H), 7.01 (dd, J = 8.8, 2.3 Hz, 1H), 5.65 (s, 2H), 3.81 (s, 3H), 2.84 (t, J = 10.9 Hz, 1H), 1.65 (d, J = 10.8 Hz, 5H), 1.42 (d, J = 11.3 Hz, 2H), 1.22 (d, J = 11.8 Hz, 3H); LCMS (ESI) m/z: 382.2 [M+H]+. Example 4. Preparation of 4-cyano-N-(5-methoxy-1,3-benzothiazol-2-yl)benzamide
Figure imgf000058_0002
Preparation of 4-cyano-N-(5-methoxy-1,3-benzothiazol-2-yl)benzamide
Figure imgf000058_0003
A mixture of 6-methoxy-1,3-benzothiazol-2-amine (300 mg, 1.7 mmol), 4-cyanobenzoic acid (267 mg, 1.8 mmol) , N,N,N′,N′-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate (750 mg, 2.0 mmol), N,N-diisopropylethylamine (642 mg, 5.0 mmol) in N,N-dimethylformamide (30 mL) then the mixture was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The crude residue was purified by prep-HPLC (Waters X bridge 150*255uM column; 35-70 % acetonitrile in a 10 mM ammonium acetate solution in water, 12 min gradient). Compound 4-cyano-N-(5- methoxy-1,3-benzothiazol-2-yl)benzamide (320 mg, 1.0 mmol, 60 %) was obtained as a pale yellow solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ = 13.15 (br s, 1H), 8.26 (d, J=8.2 Hz, 2H), 8.05 (d, J=8.1 Hz, 2H), 7.89 (d, J=8.7 Hz, 1H), 7.30 (br s, 1H), 7.00 (dd, J=2.1, 8.7 Hz, 1H), 3.85 (s, 3H); LCMS (ESI) m/z: 310.0 [M+H]+. Example 5. Preparation of N-benzyl-4-cyano-N-(5-methoxy-1,3-benzothiazol-2-yl)benzamide
Figure imgf000058_0004
Step 1: Preparation of N-benzyl-5-methoxybenzo[d]thiazol-2-amine
Figure imgf000059_0001
To a solution of benzaldehyde (0.589 g, 5.6 mmol), 5-methoxybenzo[d]thiazol-2-amine (1.00 g, 5.6 mmol) in toluene (10 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.06 g, 28 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-benzyl-5-methoxybenzo[d]thiazol-2-amine (0.500 g, 1.9 mmol, 33 %) as a white solid. LCMS (ESI) m/z: 271.2 [M+H]+. Step 2: Preparation of N-benzyl-4-cyano-N-(5-methoxy-1,3-benzothiazol-2-yl)benzamide
Figure imgf000059_0002
To a solution of 4-cyanobenzoic acid (150 mg, 1.0 mmol) in dichloromethane (2 mL) was added thionyl chloride (10 mL) at room temperature. The mixture was heated to 100 °C and stirred for 2 hours. Then the solvent was removed under the reduced pressure. The residue was dissolved in dichloromethane (2 mL) and slowly added to a solution of N-benzyl-5-methoxybenzo[d]thiazol-2-amine (250 mg, 0.93 mmol), N,N-diisopropylethylamine (359 mg, 2.8 mmol) in dichloromethane (10 mL). Then the residue was stirred at room temperature for 2 hours. The residue was concentrated under reduced pressure. Crude product was purified by silica gel column using a gradient of 0-50% ethyl acetate in petroleum ether, then recrystallized by ethanol to offer N-benzyl-4-cyano-N-(5-methoxybenzo[d]thiazol-2-yl)benzamide (72.6 mg, 0.18 mmol, 20 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 7.92 (dd, J = 8.4, 5.1 Hz, 3H), 7.70 (d, J = 8.2 Hz, 2H), 7.43 – 7.16 (m, 4H), 7.16 – 6.96 (m, 3H), 5.41 (s, 2H), 3.80 (s, 3H); LCMS (ESI) m/z: 400.0 [M+H]+. Example 6. Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)benzamide
Figure imgf000059_0003
Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)benzamide
Figure imgf000059_0004
A mixture of 6-ethyl-1,3-benzothiazol-2-amine (50.0 mg, 0.28 mmol), 4-cyanobenzoic acid (45.0 mg, 0.31 mmol), N,N,N′,N′-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate (127 mg, 0.34 mmol), N,N-diisopropylethylamine (108 mg, 0.84 mmol) in N,N-dimethylformamide (1.5 mL) then the mixture was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. Water (20 mL) was added, then the organic layer was extracted with 2x 20 mL washes of ethyl acetate. The organic layers were pooled and concentrated under reduced pressure. The crude residue was purified by prep-HPLC (Waters X bridge 150*255uM column; 35-70 % acetonitrile in an a 10mM ammonium acetate solution in water, 12 min gradient). Compound 4-cyano-N-(6-ethyl-1,3-benzothiazol-2- yl)benzamide (39.0 mg, 0.13 mmol, 45 %) was obtained as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ = 8.26 (d, J=8.2 Hz, 2H), 8.05 (d, J=8.3 Hz, 2H), 7.85 (s, 1H), 7.70 (br d, J=8.2 Hz, 1H), 7.34 (dd, J=1.0, 8.3 Hz, 1H), 2.79 - 2.67 (m, 2H), 1.25 (t, J=7.6 Hz, 3H); LCMS (ESI) m/z: 308.0 [M+H]+. Example 7. Preparation of N-benzyl-4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)benzamide
Figure imgf000060_0001
Step 1: Preparation of N-benzyl-6-ethylbenzo[d]thiazol-2-amine
Figure imgf000060_0002
To a solution of benzaldehyde (0.890 g, 8.4 mmol), 6-ethylbenzo[d]thiazol-2-amine (1.00 g, 5.6 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (0.638 g, 17 mmol) in ethanol (5 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-benzyl-6-ethylbenzo[d]thiazol-2-amine (1.00 g, 3.7 mmol, 66 %) as a white solid. LCMS (ESI) m/z: 269.1 [M+H]+. Step 2: Preparation of N-benzyl-4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)benzamide
Figure imgf000060_0003
To a solution of N-benzyl-6-ethylbenzo[d]thiazol-2-amine (268 mg, 1.0 mmol), 4-cyanobenzoic acid (147 mg, 1.0 mmol), triethylamine (303 mg, 3.0 mmol) in dichloromethane (5 mL) was added N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (570 mg, 1.5 mmol) at room temperature for 16 hours. The solvent was removed under the reduced pressure and the residue was purified by silica gel column using a gradient of 0-50% ethyl acetate in petroleum ether, then recrystallized from ethanol to offer N-benzyl-4-cyano-N-(6- ethylbenzo[d]thiazol-2-yl)benzamide (72.0 mg, 0.19 mmol, 49 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.03 – 7.82 (m, 3H), 7.74 – 7.60 (m, 3H), 7.36 – 7.15 (m, 4H), 7.07 (d, J = 7.1 Hz, 2H), 5.40 (s, 2H), 2.75 – 2.72 (m, 2H), 1.23 (t, J = 7.5 Hz, 3H); LCMS (ESI) m/z: 398.1 [M+H]+. Example 8. Preparation of 4-cyano-N-(5-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
Figure imgf000061_0001
Step 1: Preparation of 5-methoxy-N-(3-pyridylmethyl)-1,3-benzothiazol-2-amine
Figure imgf000061_0002
To a solution of 5-methoxy-1,3-benzothiazol-2-amine (500 mg, 2.8 mmol) in N,N- dimethylformamide (5 mL) was added 3-(chloromethyl)pyridine (683 mg, 4.2 mmol) and potassium carbonate (767 mg, 5.6 mmol). The reaction mixture was stirred at 90 °C for 12 hours. The product was indicated present via UPLC analysis. Water (10 mL) was added to the reaction, then the organic layer was extracted with 2x 10 mL portions of ethyl acetate. The combined organic layers were concentrated under reduced pressure. Crude product was purified by flash column chromatography using a gradient of 80-100% ethyl acetate in petroleum ether through 10 gram silica gel. Product 5-methoxy-N-(3- pyridylmethyl)-1,3-benzothiazol-2-amine (230 mg, 0.85 mmol, 31 %) was obtained as a yellow solid.1H NMR (400 MHz, Chloroform-d) δ 8.57 (d, J = 1.8 Hz, 1H), 8.47 (dd, J = 1.5, 4.8 Hz, 1H), 7.73 - 7.63 (m, 1H), 7.35 (d, J = 8.6 Hz, 1H), 7.26 - 7.15 (m, 1H), 7.01 (d, J = 2.4 Hz, 1H), 6.66 (dd, J = 2.5, 8.6 Hz, 1H), 4.60 (s, 2H), 3.75 (s, 3H). Step 2: Preparation of 4-cyano-N-(5-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000061_0003
To a solution of 5-methoxy-N-(3-pyridylmethyl)-1,3-benzothiazol-2-amine (180 mg, 0.66 mmol) in dichloromethane (3 mL) at 0 °C was added triethylamine (201 mg, 2.0 mmol) and 4-cyanobenzoyl chloride (165 mg, 1.0 mmol). The mixture was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The reaction mixture was filtered over Celite and washed with 2x 20 mL portions of ethyl acetate. The filtrate was concentrated under reduced pressure. Crude product was purified by prep-HPLC (Waters Xbridge Prep OBD C18150*40mm*10um column; 35-65 % acetonitrile in a 10 mM ammonium bicarbonate solution in water, 11 min gradient). Product 4-cyano-N-(5-methoxy-1,3- benzothiazol-2-yl)-N-(3-pyridylmethyl)benzamide (63.0 mg, 0.16 mmol, 24 %) was obtained as an off- white solid.1H NMR (400 MHz, Chloroform-d) δ = 8.53 (dd, J = 1.5, 4.8 Hz, 1H), 8.40 (d, J = 1.3 Hz, 1H), 7.75 - 7.64 (m, 3H), 7.60 - 7.48 (m, 3H), 7.32 (d, J = 2.4 Hz, 1H), 7.23 (dd, J = 4.8, 7.9 Hz, 1H), 7.02 (dd, J = 2.4, 8.8 Hz, 1H), 5.47 (s, 2H), 3.88 (s, 3H); LCMS (ESI) m/z: 401.0 [M+H]+. Example 9. Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)-2,3-dihydro-1,4- benzodioxine-6-carboxamide
Figure imgf000062_0001
Step 1: Preparation of 2,3-dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride
Figure imgf000062_0002
A mixture of 2,3-dihydrobenzo[b][1,4]dioxine-6-carboxylic acid (900 mg, 5.0 mmol) in thionyl chloride (10 mL) was stirred at 50 °C for 2 hours, then concentrated under reduced pressure. The resulting solid was 2,3-dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride (890 mg, 4.5 mmol, 90 %) as an off-white powder; LCMS (ESI) m/z: 195.1 [M+H]+. Step 2: Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)-2,3-dihydro-1,4- benzodioxine-6-carboxamide
Figure imgf000062_0003
To a solution of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (259 mg, 1.0 mmol) and triethylamine (303 mg, 3.0 mmol) in dichloromethane (5 mL) was added 2,3-dihydrobenzo[b][1,4]dioxine- 6-carbonyl chloride (398 mg, 2.0 mmol) at 0 °C under argon. The mixture was stirred at 40 °C for 4 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added, and the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude product was purified by Prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-(4-fluorobenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (12.0 mg, 0.027 mmol, 2.7 %) as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.43 (d, J = 4.0 Hz, 1H), 8.38 (s, 1H), 7.85 (d, J = 7.7 Hz, 1H), 7.58 (d, J = 8.0 Hz, 1H), 7.43 – 7.25 (m, 3H), 7.17 (d, J = 1.9 Hz, 1H), 7.09 (dd, J = 8.3, 1.9 Hz, 1H), 6.96 (d, J = 8.3 Hz, 1H), 5.51 (s, 2H), 4.29 (dd, J = 11.9, 4.7 Hz, 4H); LCMS (ESI) m/z: 422.0 [M+H]+. Example 10. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3- yl)methyl]propenamide Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3-yl)methyl]propanamide
Figure imgf000063_0001
To a solution of 6-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (425 mg, 1.6 mmol), 3- phenoxypropanoic acid (300 mg, 1.8 mmol), N,N-diisopropylethylamine (637 mg, 4.9 mmol) in tetrahydrofuran (20 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (937 mg, 2.5 mmol). The mixture was heated to 90 °C and stirred for 4 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by silica gel column using a gradient of 0-50% ethyl acetate in petroleum ether, then recrystallized with ethanol to offer N-(6- fluorobenzo[d]thiazol-2-yl)-3-phenoxy-N-(pyridin-3-ylmethyl)propanamide (125 mg, 0.31 mmol, 19 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.56 (d, J = 40.5 Hz, 2H), 7.93 (d, J = 7.2 Hz, 1H), 7.87 – 7.63 (m, 2H), 7.31 (t, J = 23.4 Hz, 4H), 6.91 (d, J = 7.3 Hz, 3H), 5.68 (s, 2H), 4.31 (s, 2H), 3.22 (s, 2H); LCMS (ESI) m/z: 408.1 [M+H]+. Example 11. Preparation of N-(4-chloro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000063_0002
Step 1: Preparation of N-benzyl-5-methoxybenzo[d]thiazol-2-amine
Figure imgf000063_0003
To a solution of nicotinaldehyde (0.594 g, 5.6 mmol), 4-chlorobenzo[d]thiazol-2-amine (1.02 g, 5.6 mmol) in toluene (10 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.06 g, 28 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-benzyl-5-methoxybenzo[d]thiazol-2-amine (0.550 g, 2.0 mmol, 36 %) as a white solid. LCMS (ESI) m/z: 276.1 [M+H]+. Step 2: Preparation of N-(4-chloro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3-yl)methyl]propanamide
Figure imgf000064_0001
To a solution of 4-chloro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (226 mg, 0.82 mmol), 3- phenoxypropanoic acid (150 mg, 0.90 mmol), N,N-diisopropylethylamine (318 mg, 2.5 mmol) in tetrahydrofuran (20 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (468 mg, 1.2 mmol). The mixture was heated to 90 °C and stirred for 4 hours. The solvent was removed under the reduced pressure and the residue was further purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(4-chlorobenzo[d]thiazol-2-yl)-3-phenoxy- N-(pyridin-3-ylmethyl)propanamide (22.2 mg, 0.052 mmol, 6.0 %) as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.70 (d, J = 1.7 Hz, 1H), 8.51 (d, J = 3.4 Hz, 1H), 7.99 (d, J = 7.3 Hz, 1H), 7.80 (d, J = 7.9 Hz, 1H), 7.54 (d, J = 7.1 Hz, 1H), 7.46 – 7.22 (m, 4H), 6.94 (t, J = 8.3 Hz, 3H), 5.69 (s, 2H), 4.34 (t, J = 5.8 Hz, 2H), 3.30 (d, J = 5.9 Hz, 2H); LCMS (ESI) m/z: 424.0 [M+H]+. Example 12. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000064_0002
Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide
Figure imgf000064_0003
To a solution of 6-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (150 mg, 0.58 mmol), N,N- diisopropylethylamine (224 mg, 1.7 mmol) in dichloromethane (10 mL) was added cyclohexylcarbonyl chloride (89.0 mg, 0.64 mmol). The mixture was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure. The residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(6-fluorobenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide (78.2 mg, 0.21 mmol, 37 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.58 (d, J = 1.8 Hz, 1H), 8.50 (dd, J = 4.8, 1.5 Hz, 1H), 7.91 (dd, J = 8.7, 2.7 Hz, 1H), 7.76 (dd, J = 8.9, 4.8 Hz, 1H), 7.71 – 7.58 (m, 1H), 7.37 (m, 1H), 7.28 (td, J = 9.1, 2.7 Hz, 1H), 5.68 (s, 2H), 2.87 (t, J = 11.3 Hz, 1H), 1.76 – 1.56 (m, 5H), 1.43 (dd, J = 22.4, 10.8 Hz, 2H), 1.23 (d, J = 13.1 Hz, 3H); LCMS (ESI) m/z: 370.1 [M+H]+. Example 13. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide
Figure imgf000065_0001
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide
Figure imgf000065_0002
To a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (1.34 g, 5.0 mmol), cyclohexanecarboxylic acid (0.640 g, 5.0 mmol), triethylamine (1.20 g, 12 mmol) in dichloromethane (50 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (2.28 g, 6.0 mmol) at room temperature for 16 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by flash column chromatography through silica gel using a gradient of 0-50% ethyl acetate in petroleum ether, then recrystallizing from ethanol. Product N-(6- ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide (2.00 g, 5.3 mmol, 88 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.56 (d, J = 1.8 Hz, 1H), 8.49 (dd, J = 5.0, 1.5 Hz, 1H), 7.81 (d, J = 1.1 Hz, 1H), 7.70 – 7.58 (m, 2H), 7.36 (dd, J = 7.5, 5.0 Hz, 1H), 7.27 (dd, J = 8.0, 2.0 Hz, 1H), 5.68 (s, 2H), 2.86 (t, J = 11.2 Hz, 1H), 2.73 – 2.68 (m, 2H), 1.65 (d, J = 11.0 Hz, 5H), 1.42 (d, J = 12 Hz, 2H), 1.24 – 1.18 (m, 6H); LCMS (ESI) m/z: 380.3 [M+H]+. Example 14. Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide
Figure imgf000065_0003
Step 1: Preparation of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000065_0004
To a solution of nicotinaldehyde (1.27 g, 12 mmol) and 4-fluorobenzo[d]thiazol-2-amine (2.00 g, 12 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (2.26 g, 60 mmol) in ethanol (100 mL). The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (1.48 g, 5.7 mmol, 48 %) as an off-white powder; LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide
Figure imgf000066_0001
To a solution of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (259 mg, 1.0 mmol) and 4- (dimethylamino)pyridine (183 mg, 1.5 mmol) in pyridine (5 mL) was added cyclohexanecarbonyl chloride (221 mg, 1.5 mmol) at 0 °C under argon. The mixture was stirred at 40 °C for 12 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added to the reaction, then the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by Prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-(4-fluorobenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide (166 mg, 0.45 mmol, 45 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.59 (d, J = 1.8 Hz, 1H), 8.50 (dd, J = 4.7, 1.2 Hz, 1H), 7.84 (d, J = 7.2 Hz, 1H), 7.66 (d, J = 8.0 Hz, 1H), 7.42 – 7.23 (m, 3H), 5.71 (s, 2H), 2.91 (t, J = 11.3 Hz, 1H), 1.67 (m, 5H), 1.43 (dd, J = 22.8, 10.9 Hz, 2H), 1.31 – 1.14 (m, 3H); LCMS (ESI) m/z: 370.1 [M+H]+. Example 15. Preparation of N-[(4-cyanophenyl)methyl]-N-(6-ethyl-1,3-benzothiazol-2-yl)pyridine-3- carboxamide
Figure imgf000066_0002
Step 1: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)pyridine-3-carboxamide
Figure imgf000066_0003
In a reaction vial, 6-ethyl-1,3-benzothiazol-2-amine (100 mg, 0.56 mmol), pyridine-3-carboxylic acid (72.5 mg, 0.59 mmol), and N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (214 mg, 0.56 mmol) were dissolved in N,N- diisopropylethylamine (3 mL). The reaction was stirred at room temperature for 12 hours. The product was indicated present via UPLC analysis. The reaction was diluted with 10 mL ethyl acetate and 10 mL deionized water. The organic layer was separated and concentrated under reduced pressure. Crude product was purified via flash column chromatography, eluting with 0-100% ethyl acetate in dichloromethane through 12 g of silica gel. Product N-(6-ethyl-1,3-benzothiazol-2-yl)pyridine-3- carboxamide (115 mg, 0.41 mmol, 73 %) was afforded as an orange solid. Step 2: Preparation of N-[(4-cyanophenyl)methyl]-N-(6-ethyl-1,3-benzothiazol-2-yl)pyridine-3- carboxamide
Figure imgf000067_0001
In a reaction vial, N-(6-ethyl-1,3-benzothiazol-2-yl)pyridine-3-carboxamide (91.0 mg, 0.32 mmol) was suspended in dimethylformamide (2 mL) with stirring.4-(bromomethyl)benzonitrile (62.9 mg, 0.32 mmol) and dipotassium carbonate (53.2 mg, 0.39 mmol) were added and the reaction was stirred at 65°C overnight. The product was indicated via UPLC analysis. The reaction was cooled to room temperature and diluted with 20 mL ethyl acetate. The solution was washed with 3x 10 mL portions of water and 1x 10 mL portion of saturated brine. The organic layer was dried over sodium sulfate, filtered, and concentrated under reduced pressure. Crude product was purified via flash column chromatography, eluting with 0- 100% ethyl acetate in hexanes through 12 g of silica gel. Product N-[(4-cyanophenyl)methyl]-N-(6-ethyl- 1,3-benzothiazol-2-yl)pyridine-3-carboxamide (21.0 mg, 0.053 mmol, 17 %) was afforded as an orange solid.1H NMR (300 MHz, Chloroform-d) δ 9.64 – 9.36 (m, 1H), 8.91 – 8.64 (m, 1H), 8.53 (d, J = 7.8 Hz, 1H), 7.90 – 7.53 (m, 3H), 7.45 (d, J = 8.7 Hz, 3H), 7.22 (dd, J = 28.7, 7.7 Hz, 2H), 5.81 (s, 2H), 2.76 (d, J = 7.5 Hz, 2H), 1.28 (d, J = 7.8 Hz, 3H); LCMS (ESI) m/z: 399.3 [M+H]+. Example 16. Preparation of N-(1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000067_0002
Step 1: Preparation of N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000067_0003
To a solution of nicotinaldehyde (2.14 g, 20 mmol) and benzo[d]thiazol-2-amine (3.00 g, 20 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (3.80 g, 100 mmol) in ethanol (100 mL). The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (2.12 g, 8.8 mmol, 44 %) as an off-white powder; LCMS (ESI) m/z: 242.1 [M+H]+. Step 2: Preparation of N-(1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000068_0001
To a solution of N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (241 mg, 1.0 mmol) and 4- (dimethylamino)pyridine (183 mg, 1.5 mmol) in pyridine (5 mL) was added benzoyl chloride (210 mg, 1.5 mmol) at 0 °C under argon. The mixture was stirred at 40 °C for 12 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added to the reaction, then the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by Prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-(benzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)benzamide (197 mg, 0.57 mmol, 57 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide- d6) δ 8.42 (d, J = 4.1 Hz, 1H), 8.32 (s, 1H), 8.04 (d, J = 7.8 Hz, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.56 (t, J = 7.1 Hz, 4H), 7.53 – 7.41 (m, 3H), 7.37 (t, J = 7.5 Hz, 1H), 7.29 (dd, J = 7.6, 4.9 Hz, 1H), 5.48 (s, 2H); LCMS (ESI) m/z: 346.0 [M+H]+. Example 17. Preparation of N-(1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide
Figure imgf000068_0002
Step 1: Preparation of N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine To a solution of nicotinaldehyde
Figure imgf000068_0003
benzo[d]thiazol-2-amine (3.00 g, 20 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (3.80 g, 100 mmol) in ethanol (100 mL). The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient of 0-5% methanol in dichloromethane to offer N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (2.12 g, 8.8 mmol, 44 %) as an off-white powder; LCMS (ESI) m/z: 242.1 [M+H]+. Step 2: Preparation of N-(1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide
Figure imgf000068_0004
To a solution of N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (241 mg, 1.0 mmol) and 4- (dimethylamino)pyridine (183 mg, 1.5 mmol) in pyridine (5 mL) was added cyclohexanecarbonyl chloride (220 mg, 1.5 mmol) at 0 °C under argon. The mixture was stirred at 40 °C for 12 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added to the reaction, then the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-(benzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide (169 mg, 0.48 mmol, 48 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.58 (d, J = 1.4 Hz, 1H), 8.49 (d, J = 4.0 Hz, 1H), 8.00 (d, J = 7.9 Hz, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.64 (d, J = 7.9 Hz, 1H), 7.43 (t, J = 7.6 Hz, 1H), 7.35 (m, 2H), 5.70 (s, 2H), 2.87 (t, J = 11.3 Hz, 1H), 1.67 (m, 5H), 1.43 (dd, J = 22.2, 10.9 Hz, 2H), 1.30 – 1.10 (m, 3H); LCMS (ESI) m/z: 352.2 [M+H]+. Example 18. Preparation of 4-cyano-N-(4-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)benzamide
Figure imgf000069_0001
Step 1: Preparation of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000069_0002
To a solution of nicotinaldehyde (1.27 g, 12 mmol) and 4-fluorobenzo[d]thiazol-2-amine (2.00 g, 12 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (2.26 g, 60 mmol) in ethanol (100 mL). The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (1.48 g, 5.7 mmol, 48 %) as an off-white powder; LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of 4-cyano-N-(4-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000069_0003
To a solution of 4-cyanobenzoic acid (147 mg, 1.0 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide (5 mL) was added 4-fluoro- N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (259 mg, 1.0 mmol). The mixture was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added and the mixture was extracted with dichloromethane (2x 30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give 4-cyano-N-(4- fluorobenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (14.0 mg, 0.035 mmol, 3.5 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.43 (d, J = 3.5 Hz, 1H), 8.33 (s, 1H), 7.99 (d, J = 8.3 Hz, 2H), 7.91 (d, J = 7.8 Hz, 1H), 7.79 (d, J = 8.3 Hz, 2H), 7.58 (d, J = 7.9 Hz, 1H), 7.40 (td, J = 8.0, 4.8 Hz, 1H), 7.36 – 7.27 (m, 2H), 5.42 (s, 2H); LCMS (ESI) m/z: 389.1 [M+H]+. Example 19. Preparation of N-(1,3-benzothiazol-2-yl)-N-benzyl-4-cyanobenzamide
Figure imgf000070_0001
Step 1: Preparation of N-benzylbenzo[d]thiazol-2-amine
Figure imgf000070_0002
To a solution of benzaldehyde (1.06 g, 10 mmol) and benzo[d]thiazol-2-amine (1.50 g, 10 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.90 g, 50 mmol) in ethanol (100 mL). The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-benzylbenzo[d]thiazol-2-amine (1.13 g, 4.7 mmol, 47 %) as an off-white powder; LCMS (ESI) m/z: 241.1 [M+H]+. Step 2: Preparation of N-(1,3-benzothiazol-2-yl)-N-benzyl-4-cyanobenzamide
Figure imgf000070_0003
To a solution of 4-cyanobenzoic acid (147 mg, 1.0 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide (5 mL) was added N- benzylbenzo[d]thiazol-2-amine (240 mg, 1.0 mmol). The mixture was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added and the mixture was extracted with dichloromethane (2x 30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by Prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-(benzo[d]thiazol-2-yl)-N-benzyl-4- cyanobenzamide (41.0 mg, 0.11 mmol, 11 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.07 (d, J = 7.8 Hz, 1H), 7.94 (d, J = 8.1 Hz, 2H), 7.78 (d, J = 8.0 Hz, 1H), 7.72 (d, J = 8.1 Hz, 2H), 7.46 (t, J = 7.4 Hz, 1H), 7.38 (t, J = 7.4 Hz, 1H), 7.24 (dq, J = 13.9, 6.9 Hz, 3H), 7.08 (d, J = 7.2 Hz, 2H), 5.42 (s, 2H); LCMS (ESI) m/z: 370.1 [M+H]+. Example 20. Preparation of N-(1,3-benzothiazol-2-yl)-4-cyano-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000071_0001
Step 1: Preparation of N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000071_0002
To a solution of nicotinaldehyde (2.14 g, 20 mmol) and benzo[d]thiazol-2-amine (3.00 g, 20 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.90 g, 50 mmol) in ethanol (100 mL). The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer afford N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (2.12 g, 8.8 mmol, 44 %) as an off-white powder; LCMS (ESI) m/z: 242.1 [M+H]+. Step 2: Preparation of N-(1,3-benzothiazol-2-yl)-4-cyano-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000071_0003
To a solution of 4-cyanobenzoic acid (147 mg, 1.0 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide (5 mL) was added N- (pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (0.241 g, 1.00 mmol). The mixture was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added and the mixture was extracted with dichloromethane (2x 30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by Prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-(benzo[d]thiazol- 2-yl)-4-cyano-N-(pyridin-3-ylmethyl)benzamide (32.0 mg, 0.085 mmol, 8.5 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.43 (d, J = 3.5 Hz, 1H), 8.33 (d, J = 1.6 Hz, 1H), 8.07 (d, J = 7.4 Hz, 1H), 7.97 (d, J = 8.3 Hz, 2H), 7.78 (m, 3H), 7.58 (d, J = 8.0 Hz, 1H), 7.49 – 7.43 (m, 1H), 7.39 (dd, J = 11.1, 4.0 Hz, 1H), 7.29 (dd, J = 7.9, 4.8 Hz, 1H), 5.41 (s, 2H); LCMS (ESI) m/z: 371.1 [M+H]+. Example 21. Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000072_0001
Step 1: Preparation of 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000072_0002
To a solution of nicotinaldehyde (1.19 g, 11 mmol), 6-methoxybenzo[d]thiazol-2-amine (2.00 g, 11 mmol) in toluene (10 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (2.11 g, 56 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol- 2-amine (1.10 g, 4.1 mmol, 36 %) as a white solid. LCMS (ESI) m/z: 272.1 [M+H]+. Step 2: Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000072_0003
To a solution of 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (271 mg, 1.0 mmol) and 4-(dimethylamino)pyridine (183 mg, 1.5 mmol) in pyridine (5 mL) was added benzoyl chloride (210 mg, 1.5 mmol) at 0 °C under argon. The mixture was stirred at 40 °C for 12 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added to the reaction, then the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by Prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-(6-methoxybenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)benzamide (67.0 mg, 0.18 mmol, 18 %) as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide- d6) δ 8.42 (d, J = 3.7 Hz, 1H), 8.31 (s, 1H), 7.67 (d, J = 8.9 Hz, 1H), 7.62 (d, J = 2.5 Hz, 1H), 7.55 (m, 4H), 7.51 – 7.44 (m, 2H), 7.29 (dd, J = 7.8, 4.8 Hz, 1H), 7.04 (dd, J = 8.9, 2.6 Hz, 1H), 5.43 (s, 2H), 3.82 (s, 3H); LCMS (ESI) m/z: 376.0 [M+H]+. Example 22. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000073_0001
To a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (105 mg, 0.39 mmol) and triethylamine (78.0 mg, 0.78 mmol) in dichloromethane (5 mL) was added benzoyl chloride (82.0 mg, 0.59 mmol) in dichloromethane (2 mL) at room temperature for 1 hour. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (72.0 mg, 0.19 mmol, 49 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.42 (d, J = 4.6 Hz, 1H), 8.30 (s, 1H), 7.85 (s, 1H), 7.68 (d, J = 8.3 Hz, 1H), 7.58 – 7.52 (m, 4H), 7.51 – 7.42 (m, 2H), 7.29 (dd, J = 10.0, 6.0 Hz, 2H), 5.45 (s, 2H), 2.75 – 2.71 (m, 2H), 1.23 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 374.1 [M+H]+. Example 23. Preparation of N-benzyl-4-cyano-N-(6-methoxy-1,3-benzothiazol-2-yl)benzamide
Figure imgf000073_0002
Step 1: Preparation of N-benzyl-6-methoxybenzo[d]thiazol-2-amine
Figure imgf000073_0003
To a solution of benzaldehyde (0.590 g, 5.6 mmol), 6-methoxybenzo[d]thiazol-2-amine (1.00 g, 5.6 mmol) in toluene (10 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.06 g, 28 mmol) in ethanol (100 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-benzyl-6-methoxybenzo[d]thiazol-2-amine (0.540 g, 2.0 mmol, 36 %) as an off-white powder; LCMS (ESI) m/z: 271.1 [M+H]+. Step 2: Preparation of N-benzyl-4-cyano-N-(6-methoxy-1,3-benzothiazol-2-yl)benzamide
Figure imgf000074_0001
To a solution of 4-cyanobenzoic acid (147 mg, 1.0 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide (5 mL) was added N- benzyl-6-methoxybenzo[d]thiazol-2-amine (270 mg, 1.0 mmol). The mixture was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added and the mixture was extracted with dichloromethane (2x 30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-benzyl-4-cyano- N-(6-methoxybenzo[d]thiazol-2-yl)benzamide (73.0 mg, 0.18 mmol, 18 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 7.93 (d, J = 8.3 Hz, 2H), 7.73 – 7.62 (m, 4H), 7.30 – 7.18 (m, 3H), 7.12 – 7.01 (m, 3H), 5.38 (s, 2H), 3.82 (s, 3H); LCMS (ESI) m/z: 400.0 [M+H]+. Example 24. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)-2,3-dihydro-1,4- benzodioxine-6-carboxamide
Figure imgf000074_0002
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)-2,3-dihydro-1,4-benzodioxine-6- carboxamide
Figure imgf000074_0003
To a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (105 mg, 0.39 mmol) and N,N-diisopropylethylamine (100 mg, 0.78 mmol) in dichloromethane (5 mL) was added 2,3- dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride (85.0 mg, 0.59 mmol) in dichloromethane (2 mL) at room temperature for 1 hour. Then the solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5 μm 4.6×50 mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (50.0 mg, 0.12 mmol, 30 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.42 (d, J = 3.6 Hz, 1H), 8.38 (s, 1H), 7.82 (s, 1H), 7.66 (d, J = 8.3 Hz, 1H), 7.57 (d, J = 7.9 Hz, 1H), 7.35 – 7.23 (m, 2H), 7.13 (d, J = 1.9 Hz, 1H), 7.05 (dd, J = 8.4, 2.0 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 5.47 (s, 2H), 4.28 (dd, J = 12.0, 4.9 Hz, 4H), 2.73 – 2.68 (m, 2H), 1.24 – 1.20 (m, 3H); LCMS (ESI) m/z: 432.1 [M+H]+. Example 25. Preparation of 4-cyano-N-(6-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)benzamide
Figure imgf000075_0001
Step 1: Preparation of 6-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000075_0002
To a solution of nicotinaldehyde (1.07 g, 10 mmol), 6-methylbenzo[d]thiazol-2-amine (1.64 g, 10 mmol) in toluene (10 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.90 g, 50 mmol) in ethanol (100 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (1.80 g, 7.0 mmol, 70 %) as a light green solid. LCMS (ESI) m/z: 256.2 [M+H]+. Step 2: Preparation of 4-cyano-N-(6-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000075_0003
A mixture of 6-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (0.255 g, 1.0 mmol), 4- cyanobenzoic acid (0.735 g, 5.0 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (2.28 g, 6.0 mmol), N,N- diisopropylethylamine (1.29 g, 10 mmol) and N,N-dimethylformamide (10 mL) was stirred at room temperature for 1 hour. The mixture was poured into water, extracted with ethyl acetate (2 x 100 mL). The combined organic phase was concentrated. The residual was purified by silica gel column chromatography using a gradient of 0-100% ethyl acetate in hexanes to afford the target compound 4- cyano-N-(6-methylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (0.0560 g, 0.15 mmol, 15 %) as an off-white solid.1H NMR (400 MHz, Chloroform-d) δ 8.53 (dd, J = 4.8, 1.6 Hz, 1H), 8.40 (s, 1H), 7.72- 7.70 (m, 3H), 7.61-7.53 (m, 4H), 7.30-7.22 (m, 2H), 5.47 (s, 2H), 2.49 (s, 3H); LCMS (ESI) m/z: 385.1 [M+H]+. Example 26. Preparation of 4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3- ylmethyl)benzamide
Figure imgf000076_0001
Step 1: Preparation of 6-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000076_0002
To a solution of nicotinaldehyde (0.956 g, 8.9 mmol), 6-fluorobenzo[d]thiazol-2-amine (1.50 g, 8.9 mmol) in toluene (10 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.70 g, 45 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (1.30 g, 5.0 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of 4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000076_0003
To a solution of 4-cyanobenzoic acid (156 mg, 1.1 mmol) in dichloromethane (2 mL) was added thionyl chloride (10 mL) at room temperature. The mixture was heated to 100 °C and stirred for 2 hours. Then the solvent was removed under the reduced pressure. The residue was dissolved in dichloromethane (2 mL) and slowly added to a solution of 6-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol- 2-amine (250 mg, 0.97 mmol), N,N-diisopropylethylamine (374 mg, 2.9 mmol) in dichloromethane (10 mL). Then the residue was stirred at room temperature for 2 hours. The residue was concentrated under reduced pressure. Crude product was purified by silica gel column using a gradient of 0-50% ethyl acetate in petroleum ether, then recrystallized by ethanol to offer 4-cyano-N-(6-fluorobenzo[d]thiazol-2- yl)-N-(pyridin-3-ylmethyl)benzamide (63.3 mg, 0.16 mmol, 17 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.43 (d, J = 3.4 Hz, 1H), 8.32 (d, J = 1.8 Hz, 1H), 8.07 – 7.95 (m, 3H), 7.87 – 7.73 (m, 3H), 7.57 (d, J = 8.0 Hz, 1H), 7.31 (m, 2H), 5.39 (s, 2H); LCMS (ESI) m/z: 388.9 [M+H]+. Example 27. Preparation of N-(6-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000076_0004
Step 1: Preparation of 6-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000077_0001
To a solution of nicotinaldehyde (1.07 g, 10 mmol), 6-methylbenzo[d]thiazol-2-amine (1.64 g, 10 mmol) in toluene (10 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.90 g, 50 mmol) in ethanol (100 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (1.80 g, 7.0 mmol, 70 %) as a light green solid. LCMS (ESI) m/z: 256.2 [M+H]+. Step 2: Preparation of N-(6-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000077_0002
To a solution of 6-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (255 mg, 1.0 mmol) and 4-(dimethylamino)pyridine (12.0 mg, 0.10 mmol) in pyridine (5 mL) was added benzoyl chloride (154 mg, 1.1 mmol) at 0 °C under argon. The mixture was stirred at 40 °C for 12 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added to the reaction, then the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The residual was purified by prep-HPLC and further washed with methanol (20 mL) to afford the target compound N-(6-methylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (95.0 mg, 0.26 mmol, 26 %) as a white solid.1H NMR (400 MHz, Chloroform-d) δ 8.49-8.48 (m, 1H), 8.36 (d, J = 1.6 Hz, 1H), 7.71 (d, J = 8.4 Hz, 1H), 7.62 (s, 1H), 7.56-7.51 (m, 2H), 7.45-7.44 (m, 4H), 7.27 (d, J = 8.4 Hz, 1H), 7.20-7.17 (m, 1H), 5.54 (s, 2H), 2.49 (s, 3H); LCMS (ESI) m/z: 360.1 [M+H]+. Example 28. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000077_0003
Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000077_0004
To a solution of 6-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (200 mg, 0.77 mmol), N,N- diisopropylethylamine (299 mg, 2.3 mmol) in dichloromethane (10 mL) was added benzoyl chloride (119 mg, 0.85 mmol). The mixture was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure. The residue was purified by silica gel column using a gradient of 0-50% ethyl acetate in petroleum ether, then recrystallized by ethanol to offer N-(6-fluorobenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (113 mg, 0.31 mmol, 40 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.42 (dd, J = 4.7, 1.3 Hz, 1H), 8.31 (d, J = 1.9 Hz, 1H), 7.96 (dd, J = 8.7, 2.6 Hz, 1H), 7.80 (dd, J = 8.9, 4.8 Hz, 1H), 7.56 (dd, J = 10.5, 5.9 Hz, 4H), 7.49 (dd, J = 9.4, 5.7 Hz, 2H), 7.30 (m, 2H), 5.45 (s, 2H); LCMS (ESI) m/z: 364.0 [M+H]+. Example 29. Preparation of N-(5-methoxy-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000078_0001
Step 1: Preparation of 5-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000078_0002
To a solution of nicotinaldehyde (1.19 g, 11 mmol), 5-methoxybenzo[d]thiazol-2-amine (2.00 g, 11 mmol) in toluene (10 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (2.11 g, 56 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 5-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol- 2-amine (1.00 g, 3.7 mmol, 33 %) as a white solid. LCMS (ESI) m/z: 272.1 [M+H]+. Step 2: Preparation of N-(5-methoxy-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000078_0003
To a solution of 5-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (250 mg, 0.92 mmol)， diisopropylethylamine (357 mg, 2.8 mmol) in dichloromethane (10 mL) was added benzoyl chloride (142 mg, 1.0 mmol). The mixture was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure. The residue was purified by HPLC (SunFire C18, 4.6*50 mm, 3.5 um column Xbridge C183.5 μm 4.6×50 mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(6-fluorobenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)benzamide (40.8 mg, 0.11 mmol, 12 %) as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide- d6) δ 8.43 (dd, J = 4.7, 1.2 Hz, 1H), 8.31 (d, J = 1.8 Hz, 1H), 7.90 (d, J = 8.7 Hz, 1H), 7.62 – 7.53 (m, 4H), 7.52 – 7.45 (m, 2H), 7.40 – 7.25 (m, 2H), 7.00 (dd, J = 8.7, 2.4 Hz, 1H), 5.46 (s, 2H), 3.80 (s, 3H); LCMS (ESI) m/z: 376.0 [M+H]+. Example 30. Preparation of N-(4-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000079_0001
Step 1: Preparation of 4-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000079_0002
To a solution of nicotinaldehyde (2.14 g, 20 mmol), 4-methylbenzo[d]thiazol-2-amine (3.28 g, 20 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (3.80 g, 100 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 4-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.80 g, 11 mmol, 55 %) as a light green solid. LCMS (ESI) m/z: 256.0 [M+H]+. Step 2: Preparation of N-(4-methyl-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide
Figure imgf000079_0003
To a solution of 4-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (255 mg, 1.0 mmol) and 4-(dimethylamino)pyridine (12.0 mg, 0.10 mmol) in pyridine (5 mL) was added benzoyl chloride (154 mg, 1.1 mmol) at 0 °C under argon. The mixture was stirred at 40 °C for 1 hour. The product was indicated present via UPLC analysis. Water (30 mL) was added to the reaction, then the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The residual was purified by silica gel column chromatography using a gradient of 0-100% ethyl acetate in hexanes, then further purified by prep-HPLC to afford the target compound N-(4-methylbenzo[d]thiazol- 2-yl)-N-(pyridin-3-ylmethyl)benzamide (39.0 mg, 0.11 mmol, 11 %) as a white solid.1H NMR (400 MHz, Chloroform-d) δ 8.48 (dd, J = 4.8, 1.2 Hz, 1H), 8.44 (d, J = 1.6 Hz, 1H), 7.76-7.45 (m, 7H), 7.26-7.18 (m, 3H), 5.56 (s, 2H), 2.62 (s, 3H); LCMS (ESI) m/z: 360.0 [M+H]+. Example 31. Preparation of N-benzyl-4-cyano-N-(6-methyl-1,3-benzothiazol-2-yl)benzamide
Figure imgf000080_0001
Step 1: Preparation of N-benzyl-6-methylbenzo[d]thiazol-2-amine
Figure imgf000080_0002
To a solution of benzaldehyde (1.06 g, 10 mmol), 6-methylbenzo[d]thiazol-2-amine (1.64 g, 10 mmol) in toluene (30 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.90 g, 50 mmol) in ethanol (10 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-benzyl-6-methylbenzo[d]thiazol-2-amine (1.00 g, 3.9 mmol, 39 %) as a white solid. LCMS (ESI) m/z: 255.1 [M+H]+. Step 2: Preparation of N-benzyl-4-cyano-N-(6-methyl-1,3-benzothiazol-2-yl)benzamide
Figure imgf000080_0003
A mixture of N-benzyl-6-methylbenzo[d]thiazol-2-amine (0.254 g, 1.0 mmol), 4-cyanobenzoic acid (0.735 g, 5.0 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (2.28 g, 6.0 mmol), N,N-diisopropylethylamine (1.29 g, 10 mmol) and N,N-dimethylformamide (8 mL) was stirred at room temperature for 1 hour. The mixture was purified directly by prep-HPLC to afford the target compound N-benzyl-4-cyano-N-(6- methylbenzo[d]thiazol-2-yl)benzamide (0.0550 g, 0.14 mmol, 14 %) as a white solid.1H NMR (400 MHz, Chloroform-d) δ 7.72-7.64 (m, 4H), 7.48 (d, J = 8.4 Hz, 1H), 7.28-7.26 (m, 1H), 7.09-7.07 (m, 2H), 5.48 (s, 2H), 2.50 (s, 3H); LCMS (ESI) m/z: 384.1 [M+H]+. Example 32. Preparation of N-benzyl-4-cyano-N-(4-methyl-1,3-benzothiazol-2-yl)benzamide
Figure imgf000080_0004
Step 1: Preparation of N-benzyl-4-methylbenzo[d]thiazol-2-amine
Figure imgf000081_0001
To a solution of 4-methylbenzo[d]thiazol-2-amine (3.28 g, 20 mmol), benzaldehyde (2.12 g, 20 mmol) in toluene (30 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (3.80 g, 100 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-benzyl-4-methylbenzo[d]thiazol-2-amine (2.70 g, 11 mmol, 53 %) as a white solid. LCMS (ESI) m/z: 255.1 [M+H]+. Step 2: Preparation of N-benzyl-4-cyano-N-(4-methyl-1,3-benzothiazol-2-yl)benzamide
Figure imgf000081_0002
A mixture of N-benzyl-4-methylbenzo[d]thiazol-2-amine (254 mg, 1.0 mmol), 4-cyanobenzoic acid (147 mg, 1.0 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (570 mg, 1.5 mmol), N,N-diisopropylethylamine (645 mg, 5.0 mmol) and N,N-dimethylformamide (6 mL) was stirred at room temperature for 1 hour. The mixture was poured into water (30 mL), extracted with ethyl acetate (50 mL x 2). The combined organic phase was concentrated under reduced pressure. The residual was purified by silica gel column chromatography using a gradient of 0-100% ethyl acetate in hexanes to afford the target compound N- benzyl-4-cyano-N-(4-methylbenzo[d]thiazol-2-yl)benzamide (39.0 mg, 0.10 mmol, 10 %) as a white solid. 1H NMR (400 MHz, Chloroform-d) δ 7.70-7.67 (m, 3H), 7.52 (d, J = 8.4 Hz, 2H), 7.28-7.26 (m, 5H), 7.14- 7.12 (m, 2H), 5.51 (s, 2H), 2.58 (s, 3H); LCMS (ESI) m/z: 384.1 [M+H]+. Example 33. Preparation of N-(5-methoxy-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)-2,3-dihydro- 1,4-benzodioxine-6-carboxamide
Figure imgf000081_0003
Preparation of N-(5-methoxy-1,3-benzothiazol-2-yl)-N-(pyridin-3-ylmethyl)-2,3-dihydro-1,4-benzodioxine- 6-carboxamide
Figure imgf000082_0001
To a solution of 2,3-dihydrobenzo[b][1,4]dioxine-6-carboxylic acid (182 mg, 1.0 mmol) in dichloromethane (2 mL) was added thionyl chloride (10 mL). The mixture was heated to 100 °C and stirred for 2 hours. The solvent was removed under the reduced pressure. The residue was dissolved in dichloromethane (2 mL) and slowly added to a solution of 5-methoxy-N-(pyridin-3- ylmethyl)benzo[d]thiazol-2-amine (250 mg, 0.92 mmol), N,N-diisopropylethylamine (357 mg, 2.8 mmol) in dichloromethane (10 mL). Then mixture was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The solvent was evaporated under reduced pressure. Crude product was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(5-methoxybenzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (139 mg, 0.32 mmol, 35 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.43 (dd, J = 4.7, 1.3 Hz, 1H), 8.39 (d, J = 1.8 Hz, 1H), 7.87 (d, J = 8.7 Hz, 1H), 7.57 (d, J = 7.9 Hz, 1H), 7.31 (dd, J = 7.3, 3.7 Hz, 2H), 7.14 (d, J = 2.0 Hz, 1H), 7.05 (dd, J = 8.3, 2.0 Hz, 1H), 7.01 – 6.83 (m, 2H), 5.49 (s, 2H), 4.28 (dd, J = 12.0, 5.0 Hz, 4H), 3.79 (s, 3H); LCMS (ESI) m/z: 434.1 [M+H]+. Example 34. Preparation of N-(5-methoxy-1,3-benzothiazol-2-yl)-3-phenoxy-N-(pyridin-3- ylmethyl)propenamide
Figure imgf000082_0002
Preparation of N-(5-methoxy-1,3-benzothiazol-2-yl)-3-phenoxy-N-(pyridin-3-ylmethyl)propanamide
Figure imgf000082_0003
To a solution of 3-phenoxypropanoic acid (168 mg, 1.0 mmol) in dichloromethane (2 mL) was added thionyl chloride (10 mL). The mixture was heated to 100 °C and stirred for 2 hours. The solvent was removed under the reduced pressure. The residue was dissolved in dichloromethane (2 mL) and slowly added to a solution of 5-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (250 mg, 0.92 mmol), N,N-diisopropylethylamine (357 mg, 2.8 mmol ) in dichloromethane (10 mL). The residue was stirred at room temperature for 2 hours. The residue was concentrated, purified by silica gel column using a gradient of 0-50% ethyl acetate in petroleum ether, then recrystallized by ethanol to offer N-(5- methoxybenzo[d]thiazol-2-yl)-3-phenoxy-N-(pyridin-3-ylmethyl)propanamide (139 mg, 0.33 mmol, 35 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.60 (s, 1H), 8.51 (d, J = 4.1 Hz, 1H), 7.86 (d, J = 8.7 Hz, 1H), 7.68 (d, J = 8.0 Hz, 1H), 7.38 (dd, J = 7.8, 4.8 Hz, 1H), 7.35 – 7.17 (m, 3H), 7.06 – 6.82 (m, 4H), 5.69 (s, 2H), 4.31 (t, J = 5.8 Hz, 2H), 3.79 (s, 3H), 3.20 (t, J = 5.8 Hz, 2H); LCMS (ESI) m/z: 420.1 [M+H]+. Example 35. Preparation of 4-cyano-N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
Figure imgf000083_0001
Step 1: Preparation of 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000083_0002
In a reaction vial, nicotinaldehyde (1.19 g, 11 mmol) and 6-methoxybenzo[d]thiazol-2-amine (2.00 g, 11 mmol) were refluxed with 4 Å molecular sieves in toluene (50 mL) under N2 overnight. The resulting bright yellow imine solution was added to a mixture of sodium borohydride (2.11 g, 56 mmol) in excess hot, dry ethanol (100 mL). The resulting mixture was cooled with stirring under N2, then concentrated under reduced pressure. The resulting solid was washed with ethanol to afford 6-methoxy-N-(pyridin-3- ylmethyl)benzo[d]thiazol-2-amine (1.32 g, 4.9 mmol, 44 %) as an off-white powder; LCMS (ESI) m/z: 272.1 [M+H]+. Step 2: Preparation of 4-cyano-N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000083_0003
To a solution of 4-cyanobenzoic acid (147 mg, 1.0 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide (5 mL) was added 6- methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (270 mg, 1.0 mmol) under argon. The mixture was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added, the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give 4-cyano-N- (6-methoxybenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (41.0 mg, 0.10 mmol, 10 %) as a white solid.1H NMR (400 MHz, Dimethylfulfoxide-d6) δ 8.43 (d, J = 3.8 Hz, 1H), 8.32 (s, 1H), 7.96 (d, J = 8.3 Hz, 2H), 7.75 (d, J = 8.3 Hz, 2H), 7.68 (d, J = 8.9 Hz, 1H), 7.64 (d, J = 2.5 Hz, 1H), 7.57 (d, J = 7.9 Hz, 1H), 7.29 (dd, J = 7.8, 4.8 Hz, 1H), 7.05 (dd, J = 8.9, 2.5 Hz, 1H), 5.37 (s, 2H), 3.82 (s, 3H); LCMS (ESI) m/z: 401.1 [M+H]+. Example 36. Preparation of 4-cyano-N-(4-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
Figure imgf000084_0001
Step 1: Preparation of 4-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000084_0002
To a solution of nicotinaldehyde (2.14 g, 20 mmol), 4-methylbenzo[d]thiazol-2-amine (3.28 g, 20 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (3.80 g, 100 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 4-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.80 g, 11 mmol, 55 %) as a light green solid. LCMS (ESI) m/z: 256.0 [M+H]+. Step 2: Preparation of 4-cyano-N-(4-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000084_0003
A mixture of 4-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (0.255 g, 1.0 mmol), 4- cyanobenzoic acid (0.735 g, 5.0 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (2.85 g, 7.5 mmol), N,N- diisopropylethylamine (1.29 g, 10 mmol) and N,N-dimethylformamide (10 mL) was stirred at room temperature for 1 hour. The mixture was poured into water, extracted with ethyl acetate (2x 150 mL). The combined organic phase was concentrated under reduced pressure. The residual was purified by silica gel column chromatography using a gradient of 0-100% ethyl acetate in hexanes to afford the target compound 4-cyano-N-(4-methylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (0.135 g, 0.35 mmol, 35 %) as a light yellow solid.1H NMR (400 MHz, Chloroform-d) δ 8.53 (dd, J = 4.8, 1.6 Hz, 1H), 8.46 (s, 1H), 7.76-7.74 (m, 2H), 7.66-7.58 (m, 4H), 7.28-7.22 (m, 3H), 5.50 (s, 2H), 2.59 (s, 3H); LCMS (ESI) m/z: 385.0 [M+H]+. Example 37. Preparation of 4-cyano-N-(4-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
Figure imgf000085_0001
Step 1: Preparation of 4-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000085_0002
To a solution of nicotinaldehyde (1.19 g, 11 mmol), 4-methoxybenzo[d]thiazol-2-amine (2.00 g, 11 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (2.11 g, 56 mmol) in ethanol (100 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 4-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol- 2-amine (1.63 g, 6.0 mmol, 54 %) as an off-white powder; LCMS (ESI) m/z: 272.1 [M+H]+. Step 2: Preparation of 4-cyano-N-(4-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000085_0003
To a solution of 4-cyanobenzoic acid (147 mg, 1.0 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide (5 mL) was added 4- methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (270 mg, 1.0 mmol). The mixture was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added, the mixture was extracted with dichloromethane (2x 30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give 4-cyano-N-(4- methoxybenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (105 mg, 0.26 mmol, 26 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.43 (d, J = 4.0 Hz, 1H), 8.30 (s, 1H), 7.96 (d, J = 8.2 Hz, 2H), 7.74 (d, J = 8.2 Hz, 2H), 7.60 (d, J = 7.9 Hz, 1H), 7.54 (d, J = 7.9 Hz, 1H), 7.37 – 7.25 (m, 2H), 7.02 (d, J = 8.0 Hz, 1H), 5.42 (s, 2H), 3.87 (s, 3H); LCMS (ESI) m/z: 401.1 [M+H]+. Example 38. Preparation of N-(4-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000086_0001
Step 1: Preparation of 4-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000086_0002
To a solution of nicotinaldehyde (1.19 g, 11 mmol), 4-methoxybenzo[d]thiazol-2-amine (2.00 g, 11 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (2.11 g, 56 mmol) in ethanol (100 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 4-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol- 2-amine (1.63 g, 6.0 mmol, 54 %) as an off-white powder; LCMS (ESI) m/z: 272.1 [M+H]+. Step 2: Preparation of N-(4-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000086_0003
To a solution of 4-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (270 mg, 1.0 mmol) and 4-(dimethylamino)pyridine (183 mg, 1.5 mmol) in pyridine (5 mL) was added benzoyl chloride (210 mg, 1.5 mmol) at 0 °C under argon. The mixture was stirred at 40 °C for 12 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added and the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-(4-methoxybenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)benzamide (132 mg, 0.35 mmol, 35 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide- d6) δ 8.42 (d, J = 3.5 Hz, 1H), 8.28 (d, J = 1.7 Hz, 1H), 7.53 (m, 7H), 7.35 – 7.26 (m, 2H), 7.01 (d, J = 7.9 Hz, 1H), 5.48 (s, 2H), 3.88 (s, 3H); LCMS (ESI) m/z: 376.1 [M+H]+. Example 39. Preparation of N-benzyl-4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)benzamide
Figure imgf000087_0001
Step 1: Preparation of N-benzyl-6-fluorobenzo[d]thiazol-2-amine
Figure imgf000087_0002
To a solution of benzaldehyde (0.631 g, 6.0 mmol), 6-fluorobenzo[d]thiazol-2-amine (1.00 g, 6.0 mmol) in toluene (10 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.13 g, 30 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-benzyl-6-fluorobenzo[d]thiazol-2-amine (0.600 g, 2.3 mmol, 39 %) as a white solid. LCMS (ESI) m/z: 259.1 [M+H]+. Step 2: Preparation of N-benzyl-4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)benzamide
Figure imgf000087_0003
To a solution of 4-cyanobenzoic acid (125 mg, 0.85 mmol) in dichloromethane (2 mL) was added thionyl chloride (10 mL). The mixture was heated to 100 °C and stirred for 2 hours. The solvent was removed under the reduced pressure. The residue was dissolved in dichloromethane (2 mL) and slowly added to a solution of N-benzyl-6-fluorobenzo[d]thiazol-2-amine (200 mg, 0.78 mmol), N,N- diisopropylethylamine (300 mg, 2.3 mmol) in dichloromethane (10 mL). The residue was stirred at room temperature for 2 hours. The residue was concentrated, purified by silica gel column using a gradient of 0-50% ethyl acetate in petroleum ether, then recrystallized by ethanol to offer N-benzyl-4-cyano-N-(6- fluorobenzo[d]thiazol-2-yl)benzamide (37.5 mg, 0.097 mmol, 12 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.05 – 7.91 (m, 3H), 7.80 (dd, J = 8.9, 4.8 Hz, 1H), 7.72 (d, J = 8.4 Hz, 2H), 7.38 – 7.17 (m, 4H), 7.08 (d, J = 6.9 Hz, 2H), 5.41 (s, 2H); LCMS (ESI) m/z: 388.0 [M+H]+. Example 40. Preparation of N-benzyl-4-cyano-N-(4-methoxy-1,3-benzothiazol-2-yl)benzamide
Figure imgf000088_0001
Step 1: Preparation of N-benzyl-4-methoxybenzo[d]thiazol-2-amine
Figure imgf000088_0002
To a solution of benzaldehyde (0.590 g, 5.6 mmol), 4-methoxybenzo[d]thiazol-2-amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.06 g, 28 mmol) in ethanol (100 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-benzyl-4-methoxybenzo[d]thiazol-2-amine (0.270 g, 1.0 mmol, 18 %) as an off-white powder; LCMS (ESI) m/z: 271.0 [M+H]+. Step 2: Preparation of N-benzyl-4-cyano-N-(4-methoxy-1,3-benzothiazol-2-yl)benzamide
Figure imgf000088_0003
To a solution of 4-cyanobenzoic acid (147 mg, 1.0 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide (5 mL) was added N- benzyl-4-methoxybenzo[d]thiazol-2-amine (270 mg, 1.0 mmol). The mixture was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added, the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-benzyl-4-cyano- N-(4-methoxybenzo[d]thiazol-2-yl)benzamide (26.0 mg, 0.065 mmol, 6.5 %) as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 7.92 (d, J = 8.2 Hz, 2H), 7.67 (d, J = 8.2 Hz, 2H), 7.60 (d, J = 8.0 Hz, 1H), 7.33 (t, J = 8.0 Hz, 1H), 7.23 (m, 3H), 7.03 (m, 3H), 5.44 (s, 2H), 3.86 (s, 3H); LCMS (ESI) m/z: 400.1 [M+H]+. Example 41. Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000089_0001
Step 1: Preparation of 3-phenoxypropanoyl chloride
Figure imgf000089_0002
A mixture of 3-phenoxypropanoic acid (830 mg, 5.0 mmol) in thionyl chloride (10 mL) was stirred at 50 °C for 2 hours, then concentrated under reduced pressure. The resulting solid was 3- phenoxypropanoyl chloride (830 mg, 4.5 mmol, 90 %) as an off-white powder; LCMS (ESI) m/z: 181.1 [M+H]+. Step 2: Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3-yl)methyl]propanamide
Figure imgf000089_0003
To a solution of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (259 mg, 1.0 mmol) and N,N-diisopropylethylamine (387 mg, 3.0 mmol) in dichloromethane (5 mL) was added 3- phenoxypropanoyl chloride (370 mg, 2.0 mmol). The mixture was stirred at 0 °C for 2 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added, and the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile and water) to give N-(4-fluorobenzo[d]thiazol-2-yl)-3-phenoxy-N-(pyridin-3- ylmethyl)propanamide (75.0 mg, 0.19 mmol, 19 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.63 (d, J = 1.9 Hz, 1H), 8.51 (d, J = 3.5 Hz, 1H), 7.85 (d, J = 7.0 Hz, 1H), 7.72 (d, J = 8.0 Hz, 1H), 7.43 – 7.24 (m, 5H), 6.93 (m, 3H), 5.70 (s, 2H), 4.32 (t, J = 5.8 Hz, 2H), 3.25 (t, J = 5.9 Hz, 2H); LCMS (ESI) m/z: 408.1 [M+H]+. Example 42. Preparation of 4-cyano-N-(5-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
Figure imgf000090_0001
Step 1: Preparation of 5-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000090_0002
To a solution of nicotinaldehyde (0.946 g, 8.8 mmol), 5-methylbenzo[d]thiazol-2-amine (1.45 g, 8.8 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.68 g, 44 mmol) in ethanol (100 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 5-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (0.770 g, 3.0 mmol, 34 %) as an off-white powder; LCMS (ESI) m/z: 256.0 [M+H]+. Step 2: Preparation of 4-cyano-N-(5-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000090_0003
To a solution of 4-cyanobenzoic acid (147 mg, 1.0 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide (5 mL) was added 5- methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (255 mg, 1.0 mmol). The mixture was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added, the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give 4-cyano-N-(5- methylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (77.0 mg, 0.20 mmol, 20 %) as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.43 (d, J = 3.9 Hz, 1H), 8.34 (s, 1H), 7.97 (d, J = 8.3 Hz, 2H), 7.92 (d, J = 8.1 Hz, 1H), 7.77 (d, J = 8.3 Hz, 2H), 7.59 (d, J = 9.1 Hz, 2H), 7.30 (dd, J = 7.9, 4.8 Hz, 1H), 7.21 (d, J = 8.2 Hz, 1H), 5.39 (s, 2H), 2.41 (s, 3H); LCMS (ESI) m/z: 385.1 [M+H]+. Example 43. Preparation of N-(5-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000090_0004
Step 1: Preparation of 5-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000091_0001
To a solution of nicotinaldehyde (0.946 g, 8.8 mmol), 5-methylbenzo[d]thiazol-2-amine (1.45 g, 8.8 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.68 g, 44 mmol) in ethanol (100 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 5-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (0.770 g, 3.0 mmol, 34 %) as an off-white powder; LCMS (ESI) m/z: 256.0 [M+H]+. Step 2: Preparation of N-(5-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000091_0002
To a solution of 5-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (255 mg, 1.0 mmol) and 4-(dimethylamino)pyridine (183 mg, 1.5 mmol) in pyridine (5 mL) was added benzoyl chloride (210 mg, 1.5 mmol) at 0 °C under argon. The mixture was stirred at 40 °C for 12 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added to the reaction, then the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-(5-methylbenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)benzamide (62.0 mg, 0.17 mmol, 17 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide- d6) δ 8.42 (d, J = 3.9 Hz, 1H), 8.31 (s, 1H), 7.90 (d, J = 8.1 Hz, 1H), 7.57 (m, 5H), 7.52 – 7.44 (m, 2H), 7.29 (dd, J = 7.8, 4.8 Hz, 1H), 7.20 (d, J = 8.1 Hz, 1H), 5.45 (s, 2H), 2.41 (s, 3H); LCMS (ESI) m/z: 360.1 [M+H]+. Example 44. Preparation of N-(5-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000091_0003
Step 1: Preparation of 5-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000091_0004
To a solution of nicotinaldehyde (0.946 g, 8.8 mmol), 5-methylbenzo[d]thiazol-2-amine (1.45 g, 8.8 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.68 g, 44 mmol) in ethanol (100 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 5-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (0.770 g, 3.0 mmol, 34 %) as an off-white powder; LCMS (ESI) m/z: 256.0 [M+H]+. Step 2: Preparation of N-(5-methyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000092_0001
To a solution of 5-methyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (255 mg, 1.0 mmol) and 4-(dimethylamino)pyridine (183 mg, 1.5 mmol) in pyridine (5 mL) was added cyclohexanecarbonyl chloride (220 mg, 1.5 mmol) at 0 °C under argon. The mixture was stirred at 40 °C for 12 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added and the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-(5-methylbenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide (52.0 mg, 0.14 mmol, 14 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.57 (s, 1H), 8.49 (d, J = 3.9 Hz, 1H), 7.85 (d, J = 8.1 Hz, 1H), 7.63 (d, J = 7.8 Hz, 1H), 7.56 (s, 1H), 7.37 (dd, J = 7.8, 4.8 Hz, 1H), 7.16 (d, J = 8.1 Hz, 1H), 5.68 (s, 2H), 2.87 (t, J = 11.2 Hz, 1H), 2.40 (s, 3H), 1.66 (m, 5H), 1.42 (dd, J = 22.0, 10.8 Hz, 2H), 1.31 – 1.12 (m, 3H); LCMS (ESI) m/z: 366.1 [M+H]+. Example 45. Preparation of N-benzyl-4-cyano-N-(5-methyl-1,3-benzothiazol-2-yl)benzamide
Figure imgf000092_0002
Step 1: Preparation of N-benzyl-5-methylbenzo[d]thiazol-2-amine
Figure imgf000092_0003
To a solution of benzaldehyde (290 mg, 2.7 mmol), 5-methylbenzo[d]thiazol-2-amine (450 mg, 2.7 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (520 mg, 14 mmol) in ethanol (40 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-benzyl-5-methylbenzo[d]thiazol-2-amine (250 mg, 0.99 mmol, 36 %) as an off-white powder; LCMS (ESI) m/z: 255.1 [M+H]+. Step 2: Preparation of N-benzyl-4-cyano-N-(5-methyl-1,3-benzothiazol-2-yl)benzamide
Figure imgf000093_0001
To a solution of 4-cyanobenzoic acid (147 mg, 1.0 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol) in N,N-dimethylformamide (5 mL) was added N- benzyl-5-methylbenzo[d]thiazol-2-amine (254 mg, 1.0 mmol). The mixture was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added, the mixture was extracted with dichloromethane (30 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give N-benzyl-4-cyano- N-(5-methylbenzo[d]thiazol-2-yl)benzamide (46.0 mg, 0.11 mmol, 11 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 7.92 (m, 3H), 7.71 (d, J = 8.3 Hz, 2H), 7.59 (s, 1H), 7.30 – 7.17 (m, 4H), 7.08 (d, J = 7.1 Hz, 2H), 5.40 (s, 2H), 2.40 (s, 3H); LCMS (ESI) m/z: 384.0 [M+H]+. Example 46. Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-({4H,5H,6H-pyrrolo[1,2- b]pyrazol-3-yl}methyl)benzamide
Figure imgf000093_0002
Step 1: Preparation of N-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)methyl)-6-ethylbenzo[d]thiazol-2- amine
Figure imgf000093_0003
To a solution of 5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-3-carbaldehyde (272 mg, 2.0 mmol), 6- ethylbenzo[d]thiazol-2-amine (356 mg, 2.0 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (228 mg, 6.0 mmol) in ethanol (40 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-((5,6- dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)methyl)-6-ethylbenzo[d]thiazol-2-amine (360 mg, 1.2 mmol, 60 %) as a white solid. LCMS (ESI) m/z: 299.1 [M+H]+. Step 2: Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-({4H,5H,6H-pyrrolo[1,2-b]pyrazol-3- yl}methyl)benzamide
Figure imgf000094_0001
To a solution of N-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)methyl)-6-ethylbenzo[d]thiazol-2- amine (131 mg, 0.40 mmol), 4-cyanobenzoic acid (60.0 mg, 0.40 mmol), N,N-diisopropylethylamine (123 mg, 1.2 mmol) in dichloromethane (5 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin- 1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (228 mg, 0.60 mmol). The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer 4-cyano-N-((5,6-dihydro-4H-pyrrolo[1,2- b]pyrazol-3-yl)methyl)-N-(6-ethylbenzo[d]thiazol-2-yl)benzamide (30.1 mg, 0.070 mmol, 18 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.03 (d, J = 8.3 Hz, 2H), 7.86 (s, 1H), 7.82 – 7.76 (m, 3H), 7.35 (dd, J = 8.5, 1.5 Hz, 1H), 7.04 (s, 1H), 5.17 (s, 2H), 3.92 (t, J = 7.1 Hz, 2H), 2.77 – 2.73 (m, 2H), 2.49 – 2.43 (m, 2H), 2.43 – 2.31 (m, 2H), 1.25 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 428.1 [M+H]+. Example 47. Preparation of 4-cyano-N-(5-methoxy-1,3-benzothiazol-2-yl)-N-({4H,5H,6H-pyrrolo[1,2- b]pyrazol-3-yl}methyl)benzamide
Figure imgf000094_0002
Step 1: Preparation of N-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)methyl)-5-methoxybenzo[d]thiazol-2- amine
Figure imgf000094_0003
To a solution of 5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-3-carbaldehyde (272 mg, 20 mmol), 5- methoxybenzo[d]thiazol-2-amine (360 mg, 2.0 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (228 mg, 6.0 mmol) in ethanol (40 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-((5,6- dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)methyl)-5-methoxybenzo[d]thiazol-2-amine (400 mg, 1.3 mmol, 67 %) as a white solid. LCMS (ESI) m/z: 301.2 [M+H]+. Step 2: Preparation of 4-cyano-N-(5-methoxy-1,3-benzothiazol-2-yl)-N-({4H,5H,6H-pyrrolo[1,2-b]pyrazol- 3-yl}methyl)benzamide
Figure imgf000095_0001
To a solution of N-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)methyl)-5-methoxybenzo[d]thiazol- 2-amine (120 mg, 0.40 mmol), 4-cyanobenzoic acid (60.0 mg, 0.40 mmol), N,N-diisopropylethylamine (123 mg, 1.2 mmol) in dichloromethane (5 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5- b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (228 mg, 0.60 mmol). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer 4-cyano-N-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)methyl)-N-(5- methoxybenzo[d]thiazol-2-yl)benzamide (33.5 mg, 0.080 mmol, 20 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.03 (d, J = 8.4 Hz, 2H), 7.90 (d, J = 8.7 Hz, 1H), 7.79 (d, J = 8.4 Hz, 2H), 7.43 (d, J = 2.4 Hz, 1H), 7.06 (s, 1H), 7.02 (dd, J = 8.7, 2.4 Hz, 1H), 5.18 (s, 2H), 3.93 (t, J = 6.9 Hz, 2H), 3.84 (s, 3H), 2.44 – 2.40 (m, 4H); LCMS (ESI) m/z: 430.0 [M+H]+. Example 48. Preparation of N-(5-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000095_0002
Step 1: Preparation of 5-ethylbenzo[d]thiazol-2-amine
Figure imgf000095_0003
A mixture of 3-ethylaniline (5.00 g, 41 mmol) and potassium thiocyanate (4.00 g, 41 mmol) in glacial acetic acid (100 mL) was cooled in an ice bath and stirred for 20 min, and then bromine (6.50 g, 41 mmol) in glacial acetic acid (10 mL) was added dropwise at such a rate to keep the temperature below 10 °C throughout the addition. The reaction mixture was stirred at room temperature for 4 hours, the hydrobromide salt thus separated out was filtered, washed with acetic acid, dried, dissolved in hot water and quenched with a solution of 7 M ammonia in methanol until pH 11. The resulting precipitate was filtered, washed with water and the crude residue was purified by flash column chromatography through silica gel using a gradient of 0-50% ethyl acetate in petroleum ether. Product 5-ethylbenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol, 14 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 7.31 (d, J = 8.4 Hz, 1H), 6.58 (d, J = 2.5 Hz, 1H), 6.48 (dd, J = 8.4, 2.6 Hz, 1H), 5.74 (s, 2H), 2.74 – 2.69 (m, 2H), 1.19 (t, J = 7.5 Hz, 3H); LCMS (ESI) m/z: 179.2 [M+H]+. Step 2: Preparation of 5-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000096_0001
A mixture of 5-ethylbenzo[d]thiazol-2-amine (2.00 g, 11 mmol), nicotinaldehyde (1.80 g, 17 mmol) and 4 Å molecular sieves in toluene (20 mL) was stirred at 100 °C for 16 hours. The reaction was cooled to room temperature. Then, a solution of sodium borohydride (0.851 mg, 22 mmol) in methanol (5 mL) was added. The suspension was stirred at 60 °C for 2 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added, the mixture was extracted with ethyl acetate (2x 100 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by flash column chromatography using a gradient of 0-35% ethyl acetate in petroleum ether. Product 5-ethyl- N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (0.200 g, 0.74 mmol, 6.6 %) was afforded as a yellow oil; LCMS (ESI) m/z: 270.1 [M+H]+. Step 3: Preparation of N-(5-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000096_0002
To a solution of 5-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (105 mg, 0.39 mmol) and triethylamine (78.0 mg, 0.78 mmol) in dichloromethane (5 mL) was added benzoyl chloride (82.0 mg, 0.59 mmol) in dichloromethane (2 mL). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(5-ethylbenzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)benzamide (26.5 mg, 0.070 mmol, 18 %) as a yellow oil.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.59 (s, 1H), 8.53 (d, J = 4.9 Hz, 1H), 7.91 (d, J = 7.9 Hz, 1H), 7.59 – 7.45 (m, 2H), 7.34 (dd, J = 11.0, 7.5 Hz, 3H), 7.28 – 7.22 (m, 2H), 7.20 – 7.16 (m, 1H), 7.12 – 7.05 (m, 1H), 5.21 (s, 2H), 2.59 – 2.53 (m, 2H), 0.91 (t, J = 7.5 Hz, 3H); LCMS (ESI) m/z: 374.1 [M+H]+. Example 49. Preparation of N-benzyl-4-cyano-N-(5-ethyl-1,3-benzothiazol-2-yl)benzamide
Figure imgf000096_0003
Step 1: Preparation of 4-cyano-N-(5-ethylbenzo[d]thiazol-2-yl)benzamide
Figure imgf000096_0004
To a solution of 5-ethylbenzo[d]thiazol-2-amine (0.356 g, 2.0 mmol), 4-cyanobenzoic acid (0.294 g, 2.0 mmol), triethylamine (0.606 g, 6.0 mmol) in dichloromethane (10 mL) was added N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (1.14 g, 3.0 mmol). The reaction was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. Water (20 mL) was added, the mixture was extracted with ethyl acetate (3x 20 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by silica gel column using a gradient of 0-50% ethyl acetate in petroleum ether. Product 4-cyano-N-(5-ethylbenzo[d]thiazol-2-yl)benzamide (0.310 g, 1.0 mmol, 50 %) as a light yellow solid. LCMS (ESI) m/z: 308.0 [M+H]+. Step 2: Preparation of N-benzyl-4-cyano-N-(5-ethyl-1,3-benzothiazol-2-yl)benzamide
Figure imgf000097_0001
To a solution of 4-cyano-N-(5-ethylbenzo[d]thiazol-2-yl)benzamide (153 mg, 0.50 mmol), (bromomethyl)benzene (170 mg, 1.0 mmol), potassium carbonate (138 mg, 1.00 mmol) in acetonitrile (5 mL) was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. Water (20 mL) was added, the mixture was extracted with ethyl acetate (3x 20 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-benzyl-4-cyano-N-(5-ethylbenzo[d]thiazol-2-yl)benzamide (26.5 mg, 0.070 mmol, 18 %) as a yellow oil.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 7.76 (d, J = 8.2 Hz, 2H), 7.55 – 7.51 (m, 3H), 7.31 (d, J = 4.4 Hz, 4H), 7.25 (dd, J = 8.5, 4.0 Hz, 1H), 7.16 (dd, J = 8.4, 2.5 Hz, 1H), 7.10 (s, 1H), 5.15 (s, 2H), 2.58 – 2.53 (m, 2H), 0.90 (t, J = 7.5 Hz, 3H); LCMS (ESI) m/z: 398.1 [M+H]+. Example 50. Preparation of N-(5-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000097_0002
Preparation of N-(5-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide
Figure imgf000097_0003
To a solution of 5-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (105 mg, 0.39 mmol) and triethylamine (78.0 mg, 0.78 mmol) in dichloromethane (5 mL) was added cyclohexanecarbonyl chloride (85.0 mg, 0.59 mmol) in dichloromethane (2 mL). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C18 3.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(5- ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide (26.1 mg, 0.070 mmol, 18 %) as a yellow oil.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.44 (d, J = 4.0 Hz, 1H), 8.35 (s, 1H), 7.70 (d, J = 8.3 Hz, 1H), 7.58 (d, J = 7.7 Hz, 1H), 7.33 (dd, J = 8.0, 5.0 Hz, 1H), 7.25 (d, J = 2.1 Hz, 1H), 7.20 (dd, J = 8.0, 2.5 Hz, 1H), 4.88 (s, 2H), 2.76 – 2.73 (m, 2H), 2.18 (s, 1H), 1.65 – 1.62 (m, 4H), 1.52 (d, J = 13.0 Hz, 1H), 1.42 – 1.38 (m, 2H), 1.15 – 1.11 (m, 3H), 0.94 – 0.90 (m, 3H); LCMS (ESI) m/z: 380.3 [M+H]+. Example 51. Preparation of 4-cyano-N-(5-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
Figure imgf000098_0001
Preparation of 4-cyano-N-(5-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000098_0002
To a solution of 5-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (105 mg, 0.39 mmol) and triethylamine (78.0 mg, 0.78 mmol) in dichloromethane (5 mL) was added 4-cyanobenzoyl chloride (97.0 mg, 0.59 mmol) in dichloromethane (2 mL). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer 4-cyano-N-(5- ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (43.0 mg, 0.11 mmol, 28 %) as a colorless oil. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.54 (s, 1H), 8.48 (d, J = 4.5 Hz, 1H), 7.79 – 7.75 (m, 3H), 7.57 – 7.52 (m, 3H), 7.39 (dd, J = 8.0, 5.0 Hz, 1H), 7.20 (d, J = 8.5 Hz, 1H), 7.16 (s, 1H), 5.19 (s, 2H), 2.57 (d, J = 7.5 Hz, 2H), 0.91 (t, J = 7.5 Hz, 3H); LCMS (ESI) m/z: 399.1 [M+H]+. Example 52. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000099_0001
Step 1: Preparation of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000099_0002
To a solution of nicotinaldehyde (481 mg, 4.5 mmol), 5-fluorobenzo[d]thiazol-2-amine (500 mg, 3.0 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (342 mg, 9.0 mmol) in ethanol (10 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (600 mg, 2.3 mmol, 77 %) as a white solid. LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3-yl)methyl]propanamide
Figure imgf000099_0003
To a solution of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (100 mg, 0.39 mmol) and N,N-diisopropylethylamine (100 mg, 0.78 mmol) in dichloromethane (5 mL) was added 3- phenoxypropanoyl chloride (72.0 mg, 0.39 mmol) in dichloromethane (2 mL). The reaction was stirred at room temperature for 12 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%- 95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/water.) to offer N-(5-fluorobenzo[d]thiazol- 2-yl)-3-phenoxy-N-(pyridin-3-ylmethyl)propanamide (32.4 mg, 0.080 mmol, 20 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.60 (s, 1H), 8.50 (d, J = 4.5 Hz, 1H), 8.05 (dd, J = 8.5, 5.6 Hz, 1H), 7.69 (d, J = 7.8 Hz, 1H), 7.61 (d, J = 9.9 Hz, 1H), 7.38 (dd, J = 7.6, 4.8 Hz, 1H), 7.32 – 7.08 (m, 3H), 6.93 – 6.88 (m, 3H), 5.69 (s, 2H), 4.32 – 4.29 (m, 2H), 3.22 (t, J = 5.7 Hz, 2H); LCMS (ESI) m/z: 408.1 [M+H]+. Example 53. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000100_0001
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3-yl)methyl]propanamide
Figure imgf000100_0002
To a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (105 mg, 0.39 mmol) and N,N-diisopropylethylamine (100 mg, 0.78 mmol) in dichloromethane (5 mL) was added 3- phenoxypropanoyl chloride (72.0 mg, 0.39 mmol) in dichloromethane (2 mL). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%- 95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/water.) to offer N-(6-ethylbenzo[d]thiazol- 2-yl)-3-phenoxy-N-(pyridin-3-ylmethyl)propanamide (16.0 mg, 0.040 mmol, 10 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.59 (d, J = 2.0 Hz, 1H), 8.49 (dd, J = 5.0, 1.5 Hz, 1H), 7.83 (d, J = 1.2 Hz, 1H), 7.67 (dd, J = 8.0, 4.0 Hz, 2H), 7.37 (dd, J = 8.0, 4.5 Hz, 1H), 7.27 (dd, J = 10.8, 5.3 Hz, 3H), 6.98 – 6.84 (m, 3H), 5.68 (s, 2H), 4.33 – 4.29 (m, 2H), 3.23 – 3.18 (m, 2H), 2.78 – 2.64 (m, 2H), 1.22 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 418.2 [M+H]+. Example 54. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]thiophene-2- carboxamide
Figure imgf000100_0003
Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]thiophene-2-carboxamide
Figure imgf000100_0004
To a solution of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (101 mg, 0.39 mmol) and N,N-diisopropylethylamine (100 mg, 0.78 mmol) in dichloromethane (5 mL) was added thiophene-2- carbonyl chloride (57.0 mg, 0.39 mmol) in dichloromethane (2 mL). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/water) to offer N-(5-fluorobenzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)thiophene-2-carboxamide (47.3 mg, 0.13 mmol, 33 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.58 – 8.45 (m, 2H), 8.08 (dd, J = 8.5, 5.5 Hz, 1H), 7.99 (dd, J = 5.0, 1.5 Hz, 1H), 7.70 – 7.57 (m, 2H), 7.53 (dd, J = 4.0, 1.5 Hz, 1H), 7.35 (dd, J = 8.0, 5.0 Hz, 1H), 7.29 – 7.23 (m, 1H), 7.24 – 7.16 (m, 1H), 5.79 (s, 2H); LCMS (ESI) m/z: 370.0 [M+H]+. Example 55. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000101_0001
Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide
Figure imgf000101_0002
To a solution of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (101 mg, 0.39 mmol) and N,N-diisopropylethylamine (100 mg, 0.78 mmol) in dichloromethane (5 mL) was added cyclohexanecarbonyl chloride (72.0 mg, 0.39 mmol) in dichloromethane (2 mL). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%- 95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(5-fluorobenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide (73.0 mg, 0.20 mmol, 51 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.57 (d, J = 1.9 Hz, 1H), 8.49 (dd, J = 5.0, 1.5 Hz, 1H), 8.03 (dd, J = 8.5, 5.5 Hz, 1H), 7.67 – 7.55 (m, 2H), 7.37 (dd, J = 8.0, 4.8 Hz, 1H), 7.26 – 7.17 (m, 1H), 5.69 (s, 2H), 2.89 – 2.84 (m, 1H), 1.67 – 1.63 (m, 5H), 1.42 (d, J = 11.0 Hz, 2H), 1.23 – 1.18 (m, 3H); LCMS (ESI) m/z: 370.2 [M+H]+. Example 56. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3- yl)methyl]acetamide
Figure imgf000101_0003
Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3-yl)methyl]acetamide
Figure imgf000102_0001
To a solution of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (101 mg, 0.39 mmol) and N,N-diisopropylethylamine (100 mg, 0.78 mmol) in dichloromethane (5 mL) was added 2-phenylacetyl chloride (60.0 mg, 0.39 mmol) in dichloromethane (2 mL). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(5- fluorobenzo[d]thiazol-2-yl)-2-phenyl-N-(pyridin-3-ylmethyl)acetamide (36.0 mg, 0.090 mmol, 23 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.58 (d, J = 1.8 Hz, 1H), 8.49 (dd, J = 4.8, 1.5 Hz, 1H), 8.03 (dd, J = 8.8, 5.5 Hz, 1H), 7.65 (d, J = 8.0 Hz, 1H), 7.59 (dd, J = 10.0, 2.5 Hz, 1H), 7.41 – 7.16 (m, 7H), 5.72 (s, 2H), 4.12 (s, 2H); LCMS (ESI) m/z: 378.1 [M+H]+. Example 57. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000102_0002
Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000102_0003
To a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (101 mg, 0.39 mmol) and N,N-diisopropylethylamine (100 mg, 0.78 mmol) in dichloromethane (5 mL) was added benzoyl chloride (55.0 mg, 0.39 mmol) in dichloromethane (2 mL). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C18 3.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/water) to offer N-(5-fluorobenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl) benzamide (93.0 mg, 0.25 mmol, 66 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.42 (dd, J = 5.0, 1.5 Hz, 1H), 8.29 (d, J = 2.0 Hz, 1H), 8.09 (dd, J = 8.5, 5.5 Hz, 1H), 7.66 – 7.61 (m, 1H), 7.61 – 7.39 (m, 6H), 7.29 – 7.26 (m, 2H), 5.46 (s, 2H); LCMS (ESI) m/z: 364.0 [M+H]+. Example 58. Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000103_0001
Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3-yl)methyl]propanamide
Figure imgf000103_0002
To a solution of 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (297 mg, 1.1 mmol), 3- phenoxypropanoic acid (200 mg, 1.2 mmol), and N,N-diisopropylethylamine (424 mg, 3.3 mmol) in tetrahydrofuran (20 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (624 mg, 1.6 mmol). The mixture was stirred at 90 °C for 4 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was further purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N- (6-methoxybenzo[d]thiazol-2-yl)-3-phenoxy-N-(pyridin-3-ylmethyl)propanamide (63.0 mg, 0.15 mmol, 14 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.60 (d, J = 1.6 Hz, 1H), 8.50 (d, J = 3.6 Hz, 1H), 7.67 (t, J = 8.7 Hz, 2H), 7.59 (d, J = 2.5 Hz, 1H), 7.38 (dd, J = 7.9, 4.8 Hz, 1H), 7.28 (dd, J = 8.5, 7.5 Hz, 2H), 7.03 (dd, J = 8.9, 2.6 Hz, 1H), 6.92 (dd, J = 13.5, 7.5 Hz, 3H), 5.66 (s, 2H), 4.31 (t, J = 5.9 Hz, 2H), 3.81 (s, 3H), 3.20 (t, J = 5.9 Hz, 2H); LCMS (ESI) m/z: 420.1 [M+H]+. Example 59. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]thiophene-2- carboxamide
Figure imgf000103_0003
Step 1: Preparation of 6-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000103_0004
To a solution of nicotinaldehyde (0.956 g, 8.9 mmol), 6-fluorobenzo[d]thiazol-2-amine (1.50 g, 8.9 mmol) in toluene (10 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.70 g, 45 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (1.30 g, 5.0 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]thiophene-2-carboxamide
Figure imgf000104_0001
To a solution of 6-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (155 mg, 0.60 mmol), N,N- diisopropylethylamine (232 mg, 1.8 mmol) in dichloromethane (5 mL) was added thiophene-2-carbonyl chloride (105 mg, 0.72 mmol) at 0 °C with stirring. The reaction was warmed to room temperature over 1 hour. The product was indicated present via UPLC analysis. Water (20 mL) was added to the reaction. The organic layer was extracted with dichloromethane (2 x 30 mL), washed with brine (30 mL), dried, and concentrated. Crude product was purified by prep-HPLC to afford the desire compound N-(6- fluorobenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)thiophene-2-carboxamide (65.8 mg, 0.18 mmol, 30 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.50 (d, J = 1.8 Hz, 1H), 8.47 (dd, J = 4.8, 1.5 Hz, 1H), 7.99 (dd, J = 5.0, 1.1 Hz, 1H), 7.96 (dd, J = 8.7, 2.7 Hz, 1H), 7.78 (dd, J = 8.9, 4.8 Hz, 1H), 7.64 (d, J = 7.9 Hz, 1H), 7.52 (dd, J = 3.8, 1.1 Hz, 1H), 7.35 (dd, J = 7.9, 4.8 Hz, 1H), 7.32-7.27 (m, 1H), 7.18 (dd, J = 5.0, 3.8 Hz, 1H), 5.77 (s, 2H); LCMS (ESI) m/z: 370.0 [M+H]+. Example 60. Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]thiophene-2- carboxamide
Figure imgf000104_0002
Step 1: Preparation of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000104_0003
To a solution of nicotinaldehyde (1.27 g, 12 mmol) and 4-fluorobenzo[d]thiazol-2-amine (2.00 g, 12 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (2.26 g, 60 mmol) in ethanol (100 mL). The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (1.48 g, 5.7 mmol, 48 %) as an off-white powder; LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]thiophene-2-carboxamide
Figure imgf000105_0001
To a solution of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (200 mg, 0.77 mmol) in tetrahydrofuran (20 mL) was added sodium hydride (61.6 mg, 1.5 mmol) at 0 °C. The mixture was stirred at 0 °C for 10 minutes. Then thiophene-2-carbonyl chloride (169 mg, 1.2 mmol) was slowly added. The reaction was heated to 70 °C and stirred for 17 hours. The product was indicated present via UPLC analysis. The mixture was quenched with crushed ice, extracted with ethyl acetate (2x 10 mL). The organic layer was washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give the desired product N-(4-fluorobenzo[d]thiazol-2- yl)-N-(pyridin-3-ylmethyl)thiophene-2-carboxamide as a white solid (76.4 mg, 0.21 mmol, 27 %).1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.55 – 8.41 (m, 2H), 8.01 (dd, J = 5.0, 1.0 Hz, 1H), 7.88 (dd, J = 7.9, 0.9 Hz, 1H), 7.66 (d, J = 8.2 Hz, 1H), 7.58 (dd, J = 3.8, 1.0 Hz, 1H), 7.46 – 7.25 (m, 3H), 7.20 (dd, J = 5.0, 3.9 Hz, 1H), 5.81 (s, 2H); LCMS (ESI) m/z: 370.0 [M+H]+. Example 61. Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000105_0002
Step 1: Preparation of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000105_0003
To a solution of nicotinaldehyde (1.27 g, 12 mmol) and 4-fluorobenzo[d]thiazol-2-amine (2.00 g, 12 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (2.26 g, 60 mmol) in ethanol (100 mL). The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (1.48 g, 5.7 mmol, 48 %) as an off-white powder; LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000106_0001
To a solution of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (200 mg, 0.77 mmol) in tetrahydrofuran (20 mL) was added sodium hydride (61.6 mg, 1.5 mmol) at 0 °C. The mixture was stirred at 0 °C for 10 min. Then benzoyl chloride (161 mg, 1.2 mmol) was slowly added. The reaction was heated to 70 °C and stirred for 17 hours. The product was indicated present via UPLC analysis. The mixture was quenched with crushed ice, extracted with ethyl acetate (2x 10 mL). The organic layer was washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated. The residue was purified by prep- HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C18 21*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give the desired product N-(4-fluorobenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide as a white solid (60.6 mg, 0.17 mmol, 22 %).1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.42 (dd, J = 4.7, 1.4 Hz, 1H), 8.31 (d, J = 1.8 Hz, 1H), 7.89 (dd, J = 7.9, 1.0 Hz, 1H), 7.65 – 7.45 (m, 6H), 7.34 (dtd, J = 22.0, 7.9, 4.5 Hz, 3H), 5.48 (s, 2H); LCMS (ESI) m/z: 364.1 [M+H]+. Example 62. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3- yl)methyl]acetamide
Figure imgf000106_0002
Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3-yl)methyl]acetamide
Figure imgf000106_0003
To a solution of 6-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (207 mg, 0.80 mmol) in tetrahydrofuran (5 mL) was added sodium hydride (35.0 mg, 0.88 mmol) at 0 °C, and the reaction was stirred at 0 °C for 1 hour. Then, 2-phenylacetyl chloride (136 mg, 0.88 mmol) was added to the reaction mixture and the reaction was warmed to room temperature over 1 hour. The product was indicated present via UPLC analysis. The reaction mixture was quenched with ice water. The product was extracted with ethyl acetate (2x 30 mL), washed with brine (30 mL), dried, concentrated under reduced pressure. Crude product was purified by pre-HPLC to afford the desire compound N-(6-fluorobenzo[d]thiazol-2-yl)- 2-phenyl-N-(pyridin-3-ylmethyl)acetamide (50.0 mg, 0.13 mmol, 17 %) as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.58 (s, 1H), 8.49 (d, J = 3.8 Hz, 1H), 8.31 (s, 1H), 7.92 (dd, J = 8.8, 2.6 Hz, 1H), 7.76 (dd, J = 8.8, 4.8 Hz, 1H), 7.65 (d, J = 7.8 Hz, 1H), 7.40 – 7.24 (m, 5H), 7.24 – 7.18 (m, 2H), 5.70 (s, 2H), 4.12 (s, 2H); LCMS (ESI) m/z: 378.1 [M+H]+. Example 63. Preparation of N-(1,3-benzothiazol-2-yl)-N-[(pyridin-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000107_0001
Step 1: Preparation of N-(pyridin-4-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000107_0002
To a solution of benzo[d]thiazol-2-amine (0.800 g, 5.8 mmol) and isonicotinaldehyde (0.621 g, 5.8 mmol) in toluene (15 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.10 g, 29 mmol) in ethanol (10 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-50% ethyl acetate in petroleum ether to offer N-(pyridin-4-ylmethyl)benzo[d]thiazol-2-amine (0.600 g, 2.5 mmol, 43 %) as a yellow solid. LCMS data unavailable. Step 2: Preparation of N-(1,3-benzothiazol-2-yl)-N-[(pyridin-4-yl)methyl]cyclohexanecarboxamide
Figure imgf000107_0003
To a solution of N-(pyridin-4-ylmethyl)benzo[d]thiazol-2-amine (200 mg, 0.83 mmol) in tetrahydrofuran (20 mL) was added sodium hydride (66.4 mg, 1.7 mmol) under argon at 0 °C. The mixture was stirred at 0 °C for 10 min. Then cyclohexanecarbonyl chloride (181 mg, 1.2 mmol) was slowly added. The reaction was heated to 70 °C and stirred for 17 hours. The product was indicated present via UPLC analysis. The reaction was quenched with crushed ice, then extracted with ethyl acetate (2x 10 mL). The organic layer was washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous trifluoroacetic acid) to give the desired product N-(benzo[d]thiazol-2-yl)-N- (pyridin-4-ylmethyl)cyclohexanecarboxamide as a white solid (82.5 mg, 0.23 mmol, 28 %). 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.53 (dd, J = 4.5, 1.5 Hz, 2H), 8.00 (d, J = 7.3 Hz, 1H), 7.71 (d, J = 7.9 Hz, 1H), 7.39 (ddd, J = 24.8, 8.2, 4.1 Hz, 2H), 7.24 (d, J = 5.9 Hz, 2H), 5.69 (s, 2H), 2.84 – 2.67 (m, 1H), 1.64 (d, J = 22.7 Hz, 5H), 1.44 (dd, J = 21.5, 10.7 Hz, 2H), 1.18 (d, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 352.2 [M+H]+. Example 64. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]benzamide
Figure imgf000108_0001
Step 1: Preparation of N-(6-fluorobenzo[d]thiazol-2-yl)-1-trityl-1H-imidazole-4-carboxamide
Figure imgf000108_0002
To a solution of 6-fluorobenzo[d]thiazol-2-amine (1.68 g, 10 mmol), 1-trityl-1H-imidazole-4- carbaldehyde (3.38 g, 10 mmol) in toluene (30 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (10 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-(6-fluorobenzo[d]thiazol-2- yl)-1-trityl-1H-imidazole-4-carboxamide (1.50 g, 3.0 mmol, 31 %) as a white solid. LCMS (ESI) m/z: 491.1[M+H]+. Step 2: Preparation of N-benzoyl-N-(6-fluorobenzo[d]thiazol-2-yl)-1-trityl-1H-imidazole-4-carboxamide
Figure imgf000108_0003
To a solution of N-(6-fluorobenzo[d]thiazol-2-yl)-1-trityl-1H-imidazole-4-carboxamide (392 mg, 0.80 mmol), N,N-diisopropylethylamine (310 mg, 2.4 mmol) in dichloromethane (5 mL) was added benzoyl chloride (134 mg, 0.96 mmol) at 0 °C. The reaction was warmed to room temperature over 1 hour. The product was indicated present via UPLC analysis. Water (20 mL) was added to the reaction. The reaction mixture was extracted with dichloromethane (2x 30 mL), washed with brine (30 mL), dried, and concentrated under reduced pressure. Crude product was carried onto next step. Product N-benzoyl- N-(6-fluorobenzo[d]thiazol-2-yl)-1-trityl-1H-imidazole-4-carboxamide (450 mg, 0.76 mmol, 93 %) was afforded as a white solid. LCMS (ESI) m/z: 595.2 [M+H]+. Step 3: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]benzamide
Figure imgf000109_0001
To a solution of N-benzoyl-N-(6-fluorobenzo[d]thiazol-2-yl)-1-trityl-1H-imidazole-4-carboxamide (210 mg, 0.35 mmol) in dichloromethane (3.5 mL) was added trifluoroacetic acid (1.8 mL) at 0 °C. The reaction was warmed to room temperature over 1 hour. The product was indicated present via UPLC analysis. The reaction mixture was concentrated under reduced pressure. The obtained solid was purified by pre-HPLC to offer N-benzoyl-N-(6-fluorobenzo[d]thiazol-2-yl)-1-trityl-1H-imidazole-4-carboxamide (50.4 mg, 0.14 mmol, 41 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 13.18 (s, 1H), 8.14 (s, 1H), 8.08 (s, 1H), 7.94 (dd, J = 8.7, 2.7 Hz, 1H), 7.82 (dd, J = 8.9, 4.8 Hz, 1H), 7.73 – 7.66 (m, 2H), 7.58 (t, J = 7.3 Hz, 1H), 7.51 (t, J = 7.3 Hz, 2H), 7.31-7.26 (m, 1H), 7.15 (s, 1H), 5.32 (s, 2H); LCMS (ESI) m/z: 353.1 [M+H]+. Example 65. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000109_0002
Step 1: Preparation of N-(6-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide
Figure imgf000109_0003
To a solution of N-(6-fluorobenzo[d]thiazol-2-yl)-1-trityl-1H-imidazole-4-carboxamide (441 mg, 0.90 mmol), N,N-diisopropylethylamine (348 mg, 2.7 mmol) in dichloromethane (5 mL) was added cyclohexanecarbonyl chloride (158 mg, 1.1 mmol) at 0 °C. The reaction was warmed to room temperature over 1 hour. The product was indicated present via UPLC analysis. Water (20 mL) was added to the reaction. The reaction mixture was extracted with dichloromethane (2x 30 mL), washed with brine (30 mL), dried, and concentrated under reduced pressure. Crude product was carried forward onto next step. Product N-(6-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4-yl)methyl)cyclohexanecarboxamide (600 mg, 1.0 mmol, 111 %) was afforded as a brown oil. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000110_0001
To a solution of N-(6-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide (330 mg, 0.55 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (2.5 mL) at 0 °C. The reaction was warmed to room temperature over 1 hour. The product was indicated present via UPLC analysis. The reaction mixture was concentrated under reduced pressure. Crude product was purified by pre-HPLC to offer N-((1H-imidazol-4-yl)methyl)-N-(6- fluorobenzo[d]thiazol-2-yl)cyclohexanecarboxamide (64.1 mg, 0.18 mmol, 33 %) as a yellow solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 12.48 (s, 1H), 8.14 (s, 1H), 7.87 (dd, J = 8.7, 2.6 Hz, 1H), 7.81 (dd, J = 8.9, 4.8 Hz, 1H), 7.73 (s, 1H), 7.31-7.26 (m, 1H), 7.12 (s, 1H), 5.46 (s, 2H), 3.27 (t, J = 11.2 Hz, 1H), 1.77 (dd, J = 25.6, 12.2 Hz, 4H), 1.67 (d, J = 11.8 Hz, 1H), 1.44 (dd, J = 23.4, 10.7 Hz, 2H), 1.36- 1.18 (m, 3H); LCMS (ESI) m/z: 359.1 [M+H]+. Example 66. Preparation of 4-cyano-N-(4,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
Figure imgf000110_0002
Step 1: Preparation of 4,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000110_0003
To a solution nicotinaldehyde (1.07 g, 10 mmol) and 4,6-difluorobenzo[d]thiazol-2-amine (1.86 g, 10 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.90 g, 50 mmol) in ethanol (100 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-25% ethyl acetate in hexanes to offer 4,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (0.440 g, 1.6 mmol, 16 %) as an off-white powder; LCMS (ESI) m/z: 278.1 [M+H]+. Step 2: Preparation of 4-cyano-N-(4,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000111_0001
To a solution of 4-cyanobenzoic acid (116 mg, 0.79 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo- [4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (360 mg, 0.95 mmol) and N,N-diisopropylethylamine (153 mg, 1.2 mmol) in N,N-dimethylformamide (5 mL) was added 4,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (220 mg, 0.79 mmol). The reaction was stirred at room temperature for 16 h. The product was indicated present via UPLC analysis. Water (30 mL) was added, the mixture was extracted with dichloromethane (2x 30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by Prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C18 21*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give 4- cyano-N-(4,6-difluorobenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (69.0 mg, 0.17 mmol, 21 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.43 (d, J = 4.1 Hz, 1H), 8.32 (s, 1H), 7.99 (d, J = 8.3 Hz, 2H), 7.93 – 7.85 (m, 1H), 7.79 (d, J = 8.2 Hz, 2H), 7.58 (d, J = 8.0 Hz, 1H), 7.48 – 7.38 (m, 1H), 7.29 (dd, J = 7.8, 4.8 Hz, 1H), 5.39 (s, 2H); LCMS (ESI) m/z: 407.0 [M+H]+. Example 67. Preparation of N-(4,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000111_0002
Step 1: Preparation of 4,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000111_0003
To a solution nicotinaldehyde (1.07 g, 10 mmol) and 4,6-difluorobenzo[d]thiazol-2-amine (1.86 g, 10 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.90 g, 50 mmol) in ethanol (100 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-25% ethyl acetate in hexanes to offer 4,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (0.440 g, 1.6 mmol, 16 %) as an off-white powder; LCMS (ESI) m/z: 278.1 [M+H]+. Step 2: Preparation of N-(4,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000112_0001
To a solution of 4,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (220 mg, 0.79 mmol) and 4-(dimethylamino)pyridine (170 mg, 1.2 mmol) in pyridine (5 mL) was added cyclohexanecarbonyl chloride (140 mg, 1.2 mmol) at 0 °C under argon. The reaction was stirred at 40 °C for 12 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added, the mixture was extracted with dichloromethane (2x 30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give N-(4,6-difluorobenzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)cyclohexanecarboxamide (104 mg, 0.27 mmol, 34 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.59 (d, J = 1.7 Hz, 1H), 8.50 (d, J = 3.6 Hz, 1H), 7.82 (dd, J = 8.2, 1.9 Hz, 1H), 7.66 (d, J = 8.0 Hz, 1H), 7.37 (ddd, J = 10.9, 7.7, 3.2 Hz, 2H), 5.69 (s, 2H), 2.91 (t, J = 11.2 Hz, 1H), 1.74 – 1.56 (m, 5H), 1.42 (dd, J = 22.5, 11.2 Hz, 2H), 1.32 – 1.14 (m, 3H); LCMS (ESI) m/z: 388.1 [M+H]+. Example 68. Preparation of 3-(benzenesulfonyl)-N-(4-chloro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000112_0002
Preparation of 3-(benzenesulfonyl)-N-(4-chloro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]propanamide
Figure imgf000112_0003
To a solution of 4-chloro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (385 mg, 1.4 mmol), 3- (phenylsulfonyl)propanoic acid (300 mg, 1.4 mmol), di(tert-butyl) carbonate (458 mg, 2.1 mmol), 4- (dimethylamino)pyridine (9.00 mg, 0.070 mmol) in acetonitrile (15 mL) was added triethylamine (14.0 mg, 0.14 mmol). The reaction was stirred at 30 °C for 2 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was further purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(4-chlorobenzo[d]thiazol-2-yl)-3-(phenylsulfonyl)-N-(pyridin-3- ylmethyl)propanamide (81.0 mg, 0.17 mmol, 12 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.59 (s, 1H), 8.53 (d, J = 4.1 Hz, 1H), 7.99 (d, J = 7.9 Hz, 1H), 7.80 (d, J = 7.6 Hz, 2H), 7.77 – 7.67 (m, 2H), 7.61 (t, J = 7.7 Hz, 2H), 7.54 (d, J = 7.7 Hz, 1H), 7.44 – 7.26 (m, 2H), 5.60 (s, 2H), 3.72 (t, J = 7.0 Hz, 2H), 3.12 (t, J = 7.0 Hz, 2H); LCMS (ESI) m/z: 472.0 [M+H]+. Example 69. Preparation of 3-(benzenesulfonyl)-N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000113_0001
Preparation of 3-(benzenesulfonyl)-N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]propanamide
Figure imgf000113_0002
To a solution of 6-chloro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (321 mg, 1.2 mmol), 3- (phenylsulfonyl)propanoic acid (250 mg, 1.2 mmol), di(tert-butyl) carbonate (382 mg, 1.8 mmol), 4- (dimethylamino)pyridine (7.00 mg, 0.059 mmol) in acetonitrile (15 mL) was added triethylamine (12.0 mg, 0.12 mmol). The reaction was stirred at 30 °C for 2 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was further purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column.The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(6-chlorobenzo[d]thiazol-2-yl)-3-(phenylsulfonyl)-N-(pyridin-3- ylmethyl)propanamide (27.2 mg, 0.060 mmol, 5.0 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.52 (d, J = 5.8 Hz, 2H), 8.16 (d, J = 2.0 Hz, 1H), 7.86 – 7.67 (m, 4H), 7.59 (t, J = 7.7 Hz, 3H), 7.45 (dd, J = 8.7, 2.1 Hz, 1H), 7.38 (dd, J = 7.8, 4.8 Hz, 1H), 5.59 (s, 2H), 3.70 (t, J = 7.0 Hz, 2H), 3.05 (t, J = 6.9 Hz, 2H); LCMS (ESI) m/z: 472.0 [M+H]+. Example 70. Preparation of N-(1,3-benzothiazol-2-yl)-N-[(pyridin-2- yl)methyl]cyclohexanecarboxamide
Figure imgf000113_0003
Step 1: Preparation of N-(pyridin-2-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000114_0001
To a solution of benzo[d]thiazol-2-amine (0.800 g, 5.8 mmol) and picolinaldehyde (0.621 g, 5.8 mmol) in toluene (15 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.10 g, 29 mmol) in ethanol (10 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-35% ethyl acetate in petroleum ether to offer N-(pyridin-2-ylmethyl)benzo[d]thiazol-2-amine (0.400 g, 1.7 mmol, 29 %) as a yellow solid. LCMS data unavailable. Step 2: Preparation of N-(1,3-benzothiazol-2-yl)-N-[(pyridin-2-yl)methyl]cyclohexanecarboxamide
Figure imgf000114_0002
To a solution of N-(pyridin-2-ylmethyl)benzo[d]thiazol-2-amine (150 mg, 0.62 mmol) in tetrahydrofuran (20 mL) was added sodium hydride (49.6 mg, 1.2 mmol) under argon at 0 °C. The mixture was stirred at 0 °C for 10 minutes. Then cyclohexanecarbonyl chloride (136 mg, 0.93 mmol) was slowly added. The reaction was heated to 70 °C and stirred for 16 hours. The product was indicated present via UPLC analysis. The mixture was quenched with crushed ice, the extracted with ethyl acetate (2x 10 mL). The organic layer was washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(benzo[d]thiazol-2-yl)-N-(pyridin-2- ylmethyl)cyclohexanecarboxamide as a white solid (78.4 mg, 0.22 mmol, 36 %). 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.48 (d, J = 4.4 Hz, 1H), 7.96 (d, J = 7.6 Hz, 1H), 7.80 (td, J = 7.7, 1.7 Hz, 1H), 7.69 (d, J = 8.0 Hz, 1H), 7.47 – 7.36 (m, 2H), 7.29 (dd, J = 13.1, 6.0 Hz, 2H), 5.74 (s, 2H), 2.85 (t, J = 11.4 Hz, 1H), 1.81 – 1.58 (m, 5H), 1.45 (dd, J = 21.8, 11.0 Hz, 2H), 1.18 (s, 3H); LCMS (ESI) m/z: 352.2 [M+H]+. Example 71. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000114_0003
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-4-yl)methyl]cyclohexanecarboxamide
Figure imgf000115_0001
To a solution of 6-ethyl-N-(pyridin-4-ylmethyl)benzo[d]thiazol-2-amine (135 mg, 0.50 mmol), N,N- diisopropylethylamine (194 mg, 1.0 mmol) in dichloromethane (3 mL) was added cyclohexanecarbonyl chloride (88.0 mg, 0.60 mmol) at 0 °C. The reaction was warmed to room temperature over 1 hour. The product was indicated present via UPLC analysis. The reaction mixture was extracted with dichloromethane (2x 30 mL), washed with brine (30 mL), dried, concentrated under reduced pressure. The obtained solid was purified by pre-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston pHlex ODS 10um 21.2×250mm 120Å. The mobile phase was acetonitrile/0.1% aqueous formic acid) to afford the desired compound N-(6-ethylbenzo[d]thiazol-2- yl)-N-(pyridin-4-ylmethyl)cyclohexanecarboxamide (72.9 mg, 0.19 mmol, 38 %) as yellow solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.52 (d, J = 6.0 Hz, 2H), 7.81 (s, 1H), 7.61 (d, J = 8.3 Hz, 1H), 7.26 (dd, J = 8.4, 1.6 Hz, 1H), 7.23 (d, J = 5.9 Hz, 2H), 5.67 (s, 2H), 2.74 – 2.68 (m, 3H), 1.67 (d, J = 9.8 Hz, 4H), 1.62-1.58 (m, 1H), 1.50-1.38 (m, 2H), 1.24-1.17 (m, 6H); LCMS (ESI) m/z: 380.2 [M+H]+. Example 72. Preparation of 3-(benzenesulfonyl)-N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]propanamide
Figure imgf000115_0002
Preparation of 3-(benzenesulfonyl)-N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]propanamide
Figure imgf000115_0003
To a solution of 3-(phenylsulfonyl)propanoic acid (999 mg, 2.7 mmol) in dichloromethane (50 mL) was added N,N-dimethylformamide (0.2 mL) and oxalyl dichloride (999 mg, 8.0 mmol) at 0 °C. The reaction was warmed to room temperature over 1 hour. The reaction was concentrated under reduced pressure. The residue was added to a solution of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (101 mg, 0.39 mmol) and sodium hydride (23.0 mg, 0.59 mmol) in dichloromethane (5 mL). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(5-fluorobenzo[d]thiazol-2-yl)-3-(phenylsulfonyl)-N-(pyridin-3- ylmethyl)propanamide (40.0 mg, 0.090 mmol, 23 %) as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.52 (dd, J = 8.0, 2.5 Hz, 2H), 8.04 (dd, J = 8.5, 5.5 Hz, 1H), 7.84 – 7.69 (m, 3H), 7.61 – 7.58 (m, 4H), 7.38 (dd, J = 8.0, 5.0 Hz, 1H), 7.26 – 7.21 (m, 1H), 5.60 (s, 2H), 3.70 (t, J = 7.0 Hz, 2H), 3.04 (t, J = 7.0 Hz, 2H); LCMS (ESI) m/z: 456.0 [M+H]+. Example 73. Preparation of 3-(benzenesulfonyl)-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000116_0001
Preparation of 3-(benzenesulfonyl)-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]propanamide
Figure imgf000116_0002
To a solution of 3-(phenylsulfonyl)propanoic acid (222 mg, 1.0 mmol) in dichloromethane (10 mL) was added oxalyl chloride (260 mg, 2.1 mmol) and N,N-dimethylformamide (1 drop) and stirred at room temperature for 3 hours. The reaction mixture was concentrated, dissolved in dichloromethane (10 mL). To a solution of 6-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (180 mg, 0.69 mmol) in tetrahydrofuran (10 mL), sodium hydride (20.0 mg, 0.83 mmol) was added at 0 °C and stirred for 1 hour. Then, the above dichloromethane solution was added to the reaction mixture and stirred at room temperature for 17 hours. The reaction mixture was added to ice water, then extracted with ethyl acetate (2x 50 mL), washed with brine (30 mL), dried, concentrated. The obtained solid was purified by pre-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston pHlex ODS 10um 21.2×250mm 120Å. The mobile phase was acetonitrile/0.1% aqueous formic acid) to afford desire compound N-(6-fluorobenzo[d]thiazol-2-yl)-3-(phenylsulfonyl)-N-(pyridin-3-ylmethyl)propanamide (38.2 mg, 0.080 mmol, 12 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.60 – 8.44 (m, 2H), 7.93 (dd, J = 8.6, 2.6 Hz, 1H), 7.78-7.71 (m, 4H), 7.59 (t, J = 7.8 Hz, 3H), 7.38 (dd, J = 7.8, 4.8 Hz, 1H), 7.33 – 7.22 (m, 1H), 5.58 (s, 2H), 3.70 (t, J = 7.0 Hz, 2H), 3.04 (t, J = 7.0 Hz, 2H); LCMS (ESI) m/z: 456.1 [M+H]+. Example 74. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-2- yl)methyl]cyclohexanecarboxamide
Figure imgf000116_0003
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-2-yl)methyl]cyclohexanecarboxamide
Figure imgf000117_0001
To a solution of 6-ethyl-N-(pyridin-2-ylmethyl)benzo[d]thiazol-2-amine (160 mg, 0.60 mmol), N,N- diisopropylethylamine (232 mg, 1.8 mmol) in dichloromethane (5 mL) was added cyclohexanecarbonyl chloride (176 mg, 1.2 mmol) at 0 °C. The reaction was warmed to room temperature over 2 hours. The product was indicated present via UPLC analysis. Water (20 mL) was added to the reaction. The reaction mixture was extracted with dichloromethane (2x 20 mL), washed with brine (20 mL), dried, concentrated. The obtained solid was purified by pre-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston pHlex ODS 10um 21.2×250mm 120Å. The mobile phase was acetonitrile/0.1% aqueous formic acid) to afford the desire compound N-(6-ethyl-1,3-benzothiazol-2- yl)-N-[(pyridin-2-yl)methyl]cyclohexanecarboxamide (49.8 mg, 0.13 mmol, 22 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.48 (d, J = 4.4 Hz, 1H), 7.79 (dd, J = 7.6, 6.2 Hz, 2H), 7.59 (d, J = 8.4 Hz, 1H), 7.41 (d, J = 7.8 Hz, 1H), 7.34 – 7.16 (m, 2H), 5.71 (s, 2H), 2.84 (t, J = 11.2 Hz, 1H), 2.69 (q, J = 7.6 Hz, 2H), 1.76-1.61 (m, 5H), 1.46-1.41(m, 2H), 1.21 (t, J = 7.6 Hz, 6H); LCMS (ESI) m/z: 380.1 [M+H]+. Example 75. Preparation of 3-(benzenesulfonyl)-N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000117_0002
Step 1: Preparation of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000117_0003
To a solution of nicotinaldehyde (1.27 g, 12 mmol) and 4-fluorobenzo[d]thiazol-2-amine (2.00 g, 12 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (2.26 g, 60 mmol) in ethanol (100 mL). The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (1.48 g, 5.7 mmol, 48 %) as an off-white powder; LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of 3-(benzenesulfonyl)-N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]propanamide
Figure imgf000118_0001
To a solution of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (300 mg, 1.2 mmol) in acetonitrile (20 mL) were added 3-(phenylsulfonyl)propanoic acid (372 mg, 1.7 mmol), triethylamine (176 mg, 1.7 mmol), di(tert-butyl) carbonate (379 mg, 1.7 mmol), and 4-(dimethylamino)pyridine (212 mg, 1.7 mmol). The mixture was stirred at 50 °C for 17 hours. The product was indicated present via UPLC analysis. The mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product 3-(benzenesulfonyl)-N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]propenamide as a white solid (17.6 mg, 0.040 mmol, 3.3 %).1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.53 (d, J = 2.5 Hz, 2H), 7.88 – 7.68 (m, 4H), 7.65 – 7.55 (m, 3H), 7.41 – 7.27 (m, 3H), 5.61 (s, 2H), 3.71 (t, J = 7.0 Hz, 2H), 3.07 (t, J = 7.0 Hz, 2H); LCMS (ESI) m/z: 455.9 [M+H]+. Example 76. Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-4- yl)methyl]benzamide
Figure imgf000118_0002
Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-4-yl)methyl]benzamide
Figure imgf000118_0003
A mixture of 6-ethyl-N-(pyridin-4-ylmethyl)benzo[d]thiazol-2-amine (160 mg, 0.60 mmol), N,N- diisopropylethylamine (181 mg, 1.8 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (342 mg, 0.90 mmol), 4- cyanobenzoic acid (132 mg, 0.90 mmol) in N,N-dimethylformamide (3 mL) was and stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The reaction mixture was filtered. The filtrate was concentrated under reduced pressure and purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston pHlex ODS 10um 21.2×250mm 120Å. The mobile phase was acetonitrile/0.1% aqueous formic acid) to afford the desire compound 4-cyano-N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-4-ylmethyl)benzamide (45.5 mg, 0.11 mmol, 19 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.51 – 8.39 (m, 2H), 7.94 (d, J = 8.3 Hz, 2H), 7.89 (s, 1H), 7.74 (d, J = 8.3 Hz, 2H), 7.64 (d, J = 8.3 Hz, 1H), 7.29 (dd, J = 8.4, 1.5 Hz, 1H), 7.17 (d, J = 5.8 Hz, 2H), 5.37 (s, 2H), 2.75-2.70 (m, 2H), 1.23 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 399.1 [M+H]+. Example 77. Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-2- yl)methyl]benzamide
Figure imgf000119_0001
Step 1: Preparation of 6-ethyl-N-(pyridin-2-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000119_0002
To a solution of picolinaldehyde (1.06 g, 9.0 mmol), 6-ethylbenzo[d]thiazol-2-amine (1.60 g, 9.0 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.70 g, 45 mmol) in excess hot, dry ethanol (100 mL). The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-2-ylmethyl)benzo[d]thiazol-2- amine (1.10 g, 4.1 mmol, 46 %) as yellow solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-2-yl)methyl]benzamide
Figure imgf000119_0003
A solution of 6-ethyl-N-(pyridin-2-ylmethyl)benzo[d]thiazol-2-amine (160 mg, 0.60 mmol), 4- cyanobenzoic acid (176 mg, 1.2 mmol), N,N-diisopropylethylamine (232 mg, 1.8 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol) in N,N-dimethylformamide (3 mL) was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The reaction mixture was filtered and concentrated under reduced pressure. Crude product was purified by pre-HPLC (the crude samples were dissolved in N,N-dimethylformamide unless otherwise noted before purification. Boston pHlex ODS 10um 21.2×250mm 120Å. The mobile phase was acetonitrile/0.1% aqueous formic acid) to afford desire compound 4-cyano-N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-2-ylmethyl)benzamide (75.7 mg, 0.18 mmol, 32 %) as yellow solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.44 (d, J = 4.2 Hz, 1H), 7.92 (d, J = 8.4 Hz, 2H), 7.85 (d, J = 1.1 Hz, 1H), 7.78 (d, J = 8.5 Hz, 2H), 7.74-7.70 (m, 1H), 7.61 (d, J = 8.3 Hz, 1H), 7.32 (d, J = 7.9 Hz, 1H), 7.28-7.22 (m, 2H), 5.44 (s, 2H), 2.71 (q, J = 7.6 Hz, 2H), 1.22 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 399.1 [M+H]+. Example 78. Preparation of 4-cyano-N-(5,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
Figure imgf000120_0001
Step 1: Preparation of 5,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000120_0002
To a solution of 5,6-difluorobenzo[d]thiazol-2-amine (600 mg, 3.2 mmol), nicotinaldehyde (343 mg, 3.2 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (243 mg, 6.4 mmol) in excess hot, dry ethanol (100 mL). The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-100% ethyl acetate in hexanes to offer 5,6-difluoro-N-(pyridin-3- ylmethyl)benzo[d]thiazol-2-amine (750 mg, 2.7 mmol, 84 %) as a white solid. LCMS (ESI) m/z: 278.1 [M+H]+. Step 2: Preparation of 4-cyano-N-(5,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000120_0003
A mixture of 5,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (0.130 g, 0.57 mmol), 4- cyanobenzoic acid (0.419 g, 2.9 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (2.17 g, 5.7 mmol), N,N- diisopropylethylamine (0.349 g, 5.7 mmol) in N,N-dimethylformamide (10 mL) was stirred 90 °C for 18 hours. The product was indicated present via UPLC analysis. The mixture was poured into water, then extracted with ethyl acetate (2x 100 mL). The combined organic phase was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography using a gradient of 0-80% ethyl acetate in petroleum ether, then triturated with methanol (10 mL) to afford the target compound 4- cyano-N-(5,6-difluorobenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)benzamide (0.0324 g, 0.080 mmol, 14 %) as a grey solid.1H NMR (400 MHz, Chloroform-d) δ 8.43 (dd, J = 4.8, 1.2 Hz, 1H), 8.31 (d, J = 1.6 Hz, 1H), 8.24 (dd, J = 10.0, 8.0 Hz, 1H), 7.98 (d, J = 8.4 Hz, 2H), 7.93 (dd, J = 11.2, 7.2 Hz, 1H), 7.77 (d, J = 8.4 Hz, 2H), 7.56 (d, J = 8.4 Hz, 1H), 7.29 (dd, J = 7.6, 4.8 Hz, 1H), 5.38 (s, 2H); LCMS (ESI) m/z: 407.1 [M+H]+. Example 79. Preparation of N-(5,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000121_0001
Step 1: Preparation of 5,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000121_0002
To a solution of 5,6-difluorobenzo[d]thiazol-2-amine (600 mg, 3.2 mmol), nicotinaldehyde (343 mg, 3.2 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (243 mg, 6.4 mmol) in excess hot, dry ethanol (100 mL). The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-100% ethyl acetate in hexanes to offer 5,6-difluoro-N-(pyridin-3- ylmethyl)benzo[d]thiazol-2-amine (750 mg, 2.7 mmol, 84 %) as a white solid. LCMS (ESI) m/z: 278.1 [M+H]+. Step 2: Preparation of N-(5,6-difluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000121_0003
A mixture of 5,6-difluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (200 mg, 0.22 mmol), cyclohexanecarbonyl chloride (256 mg, 1.8 mmol), 4-(dimethylamino)pyridine (20.0 mg, 0.16 mmol) in pyridine (8 mL) was stirred at 90 °C under nitrogen atmosphere for 2 hours. The product was indicated present via UPLC analysis. The mixture was poured into water, then extracted with ethyl acetate (2x 100 mL). The combined organic phase was concentrated under reduced pressure. The resulting residual was purified by prep-HPLC to afford the target compound N-(5,6-difluorobenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide (102 mg, 0.26 mmol, 30 %) as a white solid.1H NMR (400 MHz, Chloroform-d) δ 8.57 (d, J = 1.2 Hz, 1H), 8.49 (d, J = 4.0 Hz, 1H), 8.16 (dd, J = 10.4, 8.0 Hz, 1H), 7.85 (dd, J = 11.6, 7.2 Hz, 1H), 7.62 (d, J = 8.0 Hz, 1H), 7.36 (dd, J = 7.6, 4.8 Hz, 1H), 5.68 (s, 2H), 2.90-2.84 (m, 1H), 1.66-1.63 (m, 5H), 1.46-1.37 (m, 2H), 1.27-1.15 (m, 3H); LCMS (ESI) m/z: 388.1 [M+H]+. Example 80. Preparation of N-(1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000122_0001
Step 1: Preparation of N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine
Figure imgf000122_0002
To a solution of benzo[d]thiazol-2-amine (1.10 g, 7.4 mmol), 1-trityl-1H-imidazole-4-carbaldehyde (2.50 g, 7.4 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.40 g, 37 mmol) in ethanol (7.4 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-((1-trityl-1H-imidazol-4- yl)methyl)benzo[d]thiazol-2-amine (2.20 g, 4.7 mmol, 63 %) as a yellow solid. LCMS (ESI) m/z: 473.1[M+H]+. Step 2: Preparation of N-(benzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide
Figure imgf000122_0003
To a solution of N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (800 mg, 1.7 mmol), N,N-diisopropylethylamine (660 mg, 5.1 mmol) in dichloromethane (20 mL) was added cyclohexanecarbonyl chloride (296 mg, 2.0 mmol) at 0 °C. The reaction was warmed to room temperature over 1 hour. The product was indicated present via UPLC analysis. Water (20 mL) was added to the reaction. The reaction mixture was extracted with dichloromethane (2x 30 mL), washed with brine (30 mL), dried, and concentrated under reduced pressure. Crude product was carried onto next step. Product N-(benzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4-yl)methyl)cyclohexanecarboxamide (600 mg, 1.0 mmol, 61 %) was afforded as a brown oil liquid. Step 3: Preparation of N-(1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]cyclohexanecarboxamide
Figure imgf000123_0001
To a solution of N-(benzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide (100 mg, 0.17 mmol) in dichloromethane (1 mL) was added trifluoroacetic acid (1 mL) at 0 °C. The reaction was warmed to room temperature over 1 hour. The product was indicated present via UPLC analysis. The reaction mixture was concentrated under reduced pressure. The obtained solid was purified by pre-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile/0.1% aqueous formic acid) afford desired product N-((1H-imidazol-4-yl)methyl)-N- (benzo[d]thiazol-2-yl)cyclohexanecarboxamide (15.1 mg, 0.040 mmol, 26 %) as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.15 (s, 1H), 7.97 (d, J = 8.0 Hz, 1H), 7.93 (s, 1H), 7.80 (d, J = 8.4 Hz, 1H), 7.44 (t, J = 7.6 Hz, 1H), 7.32 (t, J = 7.6 Hz, 1H), 7.21 (s, 1H), 5.51 (s, 2H), 3.32 (t, J = 11.0 Hz, 1H), 1.81 (d, J = 12.0 Hz, 2H), 1.74 (d, J = 12.4 Hz, 2H), 1.68 (d, J = 11.6 Hz, 1H), 1.50-1.41 (m, 2H), 1.34- 1.21 (m, 3H); LCMS (ESI) m/z: 341.1 [M+H]+. Example 81. Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3- yl)methyl]acetamide
Figure imgf000123_0002
Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3-yl)methyl]acetamide
Figure imgf000123_0003
To a solution of 4-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (200 mg, 0.77 mmol) in tetrahydrofuran (20 mL) was added sodium hydride (61.6 mg, 1.5 mmol) under argon at 0 °C. The reaction was stirred at 0 °C for 10 min. Then 2-phenylacetyl chloride (177 mg, 1.2 mmol) was slowly added. The reaction was warmed to room temperature with stirring over 2 hours. The mixture was quenched with crushed ice, extracted with ethyl acetate (2x 10 mL). The organic layer was washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(4-fluorobenzo[d]thiazol-2-yl)-2-phenyl-N-(pyridin-3- ylmethyl)acetamide as a white solid (82.1 mg, 0.22 mmol, 28 %).1H NMR (400 MHz, Dimethylsulfoxide- d6) δ 8.61 (d, J = 1.9 Hz, 1H), 8.50 (dd, J = 4.8, 1.5 Hz, 1H), 7.83 (dd, J = 7.8, 1.0 Hz, 1H), 7.68 (d, J = 8.0 Hz, 1H), 7.38 – 7.17 (m, 8H), 5.73 (s, 2H), 4.15 (s, 2H); LCMS (ESI) m/z: 378.1 [M+H]+. Example 82. Preparation of tert-butyl 2-{N-[(pyridin-3-yl)methyl]cyclohexaneamido}-4H,5H,6H,7H- [1,3]thiazolo[5,4-c]pyridine-5-carboxylate
Figure imgf000124_0001
Step 1: Preparation of tert-butyl 2-{[(pyridin-3-yl)methyl]amino}-4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridine- 5-carboxylate
Figure imgf000124_0002
In a reaction vial, a solution of pyridine-3-carbaldehyde (83.8 mg, 0.78 mmol) and tert-butyl 2- amino-4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridine-5-carboxylate (200 mg, 0.78 mmol) in dry toluene (7.8 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (68.0 mg, 1.8 mmol) in dry ethyl alcohol (7.8 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product tert-butyl 2-{[(pyridin-3-yl)methyl]amino}-4H,5H,6H,7H- [1,3]thiazolo[5,4-c]pyridine-5-carboxylate (188 mg, 0.55 mmol, 69 %) was afforded as an orange oil.1H NMR (300 MHz, Chloroform-d) δ 8.62 (d, J = 2.3 Hz, 1H), 8.55 (dd, J = 4.8, 1.6 Hz, 1H), 7.79 – 7.66 (m, 1H), 7.36 – 7.23 (m, 1H), 4.75 (s, 1H), 4.45 (d, J = 23.9 Hz, 4H), 3.68 (t, J = 5.6 Hz, 2H), 2.63 (t, J = 5.9 Hz, 2H), 1.47 (s, 9H); LCMS (ESI) m/z: 347.2 [M+H]+. Step 2: Preparation of tert-butyl 2-{N-[(pyridin-3-yl)methyl]cyclohexaneamido}-4H,5H,6H,7H- [1,3]thiazolo[5,4-c]pyridine-5-carboxylate
Figure imgf000125_0001
In a reaction vial, tert-butyl 2-{[(pyridin-3-yl)methyl]amino}-4H,5H,6H,7H-[1,3]thiazolo[5,4- c]pyridine-5-carboxylate (189 mg, 0.54 mmol) and dipotassium carbonate (112 mg, 0.81 mmol) were stirred magnetically in dichloromethane (1 mL) at 0 °C. To the vial, cyclohexanecarbonyl chloride (95.2 mg, 0.65 mmol) was added dropwise. The reaction was warmed up to room temperature and was stirred for 45 minutes. The starting material was indicated to be consumed by UPLC and TLC analysis.1 mL water was added to the reaction vial and the organic layer was separated with 2x 3 mL washes with dichloromethane and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product tert-butyl 2-{N-[(pyridin-3-yl)methyl]cyclohexaneamido}-4H,5H,6H,7H-[1,3]thiazolo[5,4- c]pyridine-5-carboxylate (111 mg, 0.25 mmol, 45 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.54 (d, J = 4.5 Hz, 2H), 7.52 (d, J = 4.8 Hz, 1H), 7.30 – 7.21 (m, 1H), 5.50 (s, 2H), 4.55 (s, 2H), 3.71 (t, J = 5.8 Hz, 2H), 2.80 – 2.51 (m, 3H), 1.85 – 1.52 (m, 7H), 1.48 (s, 9H), 1.32 – 1.10 (m, 3H); LCMS (ESI) m/z: 457.3 [M+H]+. Example 83. Preparation of N-{5-methyl-4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin-2-yl}-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000125_0002
Step 1: Preparation of N-[(pyridin-3-yl)methyl]-N-{4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin-2- yl}cyclohexanecarboxamide
Figure imgf000125_0003
In a reaction vial, tert-butyl 2-{N-[(pyridin-3-yl)methyl]cyclohexaneamido}-4H,5H,6H,7H- [1,3]thiazolo[5,4-c]pyridine-5-carboxylate (0.103 g, 0.22 mmol) was dissolved in dichloromethane (1 mL). A solution of 4 M hydrochloric acid in dioxane (1.0 mL, 4.0 mmol) was added to the vial and the reaction was stirred at room temperature for 2 hours. The product confirmed present via UPLC analysis and crude product N-[(pyridin-3-yl)methyl]-N-{4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin-2-yl}cyclohexanecarboxamide was assumed 100% yield and carried onto next step; LCMS (ESI) m/z: 357.2 [M+H]+. Step 2: Preparation of N-{5-methyl-4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin-2-yl}-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000126_0001
In a reaction vial, N-[(pyridin-3-yl)methyl]-N-{4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin-2- yl}cyclohexanecarboxamide (39.7 mg, 0.11 mmol) was dissolved in dichloromethane (5.7 mL). formaldehyde (18.5 mg, 0.22 mmol), acetic acid (0.013 mL, 0.22 mmol), and sodium triacetoxyborohydride (37.5 mg, 0.18 mmol) were added and the reaction was stirred at room temperature. After 5 minutes, additional sodium triacetoxyborohydride (37.5 mg, 0.18 mmol) was added and the reaction was stirred for 6 hours. The product was indicated present via UPLC analysis.1 M sodium hydroxide and saturated sodium bicarbonate were added to quench the reaction. The reaction was washed with 2x 20 mL portions of dichloromethane and the organic layers were separated and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product N-{5-methyl- 4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin-2-yl}-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide (40.1 mg, 0.11 mmol, 98 %) was afforded as a cream solid.1H NMR (300 MHz, Chloroform-d) δ 8.53 (d, J = 5.4 Hz, 2H), 7.49 (s, 1H), 7.30 – 7.16 (m, 1H), 5.51 (s, 2H), 3.59 (s, 2H), 2.77 (s, 4H), 2.65 – 2.43 (m, 4H), 1.86 – 1.45 (m, 8H), 1.36 – 1.05 (m, 6H), 0.99 – 0.75 (m, 1H); LCMS (ESI) m/z: 371.3 [M+H]+. Example 84. Preparation of N-{5-ethyl-4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin-2-yl}-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000126_0002
Step 1: Preparation of N-[(pyridin-3-yl)methyl]-N-{4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin-2- yl}cyclohexanecarboxamide
Figure imgf000126_0003
In a reaction vial, tert-butyl 2-{N-[(pyridin-3-yl)methyl]cyclohexaneamido}-4H,5H,6H,7H- [1,3]thiazolo[5,4-c]pyridine-5-carboxylate (0.103 g, 0.22 mmol) was dissolved in dichloromethane (0.22 mL). A solution of 4 M hydrochloric acid in dioxane (1.00 mL, 4.0 mmol) was added to the vial and the reaction was stirred at room temperature for 2 hours. The product confirmed present via UPLC analysis and crude product N-[(pyridin-3-yl)methyl]-N-{4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin-2- yl}cyclohexanecarboxamide was assumed 100% yield and carried onto next step; LCMS (ESI) m/z: 357.2 [M+H]+. Step 2: Preparation of N-{5-ethyl-4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin-2-yl}-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000127_0001
In a reaction vial, N-[(pyridin-3-yl)methyl]-N-{4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin-2- yl}cyclohexanecarboxamide (39.7 mg, 0.11 mmol) was dissolved in dichloromethane (5.7 mL). acetaldehyde (0.035 mL, 0.22 mmol), acetic acid (0.013 mL, 0.22 mmol), and sodium triacetoxyborohydride (37.5 mg, 0.18 mmol) were added and the reaction was stirred at room temperature. After 5 minutes, additional sodium triacetoxyborohydride (37.5 mg, 0.18 mmol) was added and the reaction was stirred for 6 hours. The product was indicated present via UPLC analysis.1 M sodium hydroxide and saturated sodium bicarbonate were added to quench the reaction. The reaction was washed with 2x 20 mL portions of dichloromethane and the organic layers were separated and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product N-{5-ethyl- 4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin-2-yl}-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide (42.6 mg, 0.11 mmol, 100 %) was afforded as an orange solid.1H NMR (300 MHz, Chloroform-d) δ 8.59 – 8.46 (m, 2H), 7.48 (s, 1H), 7.23 (dd, J = 7.7, 5.1 Hz, 1H), 5.51 (s, 2H), 3.63 (d, J = 1.8 Hz, 2H), 2.87 – 2.70 (m, 3H), 2.71 – 2.49 (m, 3H), 1.85 – 1.46 (m, 8H), 1.27 – 1.08 (m, 6H); LCMS (ESI) m/z: 385.2 [M+H]+. Example 85. Preparation of N-(6-cyano-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000127_0002
Step 1: Preparation of 2-(pyridin-3-ylmethylamino)benzo[d]thiazole-6-carbonitrile
Figure imgf000127_0003
To a solution of 2-aminobenzo[d]thiazole-6-carbonitrile (400 mg, 2.3 mmol) and nicotinaldehyde (244 mg, 2.3 mmol) in toluene (16 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (431 mg, 11 mmol) in ethanol (4 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 2-(pyridin-3- ylmethylamino)benzo[d]thiazole-6-carbonitrile as a yellow solid (220 mg, 0.83 mmol, 36 %) as a yellow solid. Step 2: Preparation of N-(6-cyano-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide
Figure imgf000128_0001
To a solution of 2-(pyridin-3-ylmethylamino)benzo[d]thiazole-6-carbonitrile (100 mg, 0.37 mmol) in dichloromethane (10 mL) was added N,N-diisopropylethylamine (143 mg, 1.1 mmol) under argon at 0 °C. The reaction was stirred at 0 °C for 10 min. Then cyclohexanecarbonyl chloride (80.3 mg, 0.55 mmol) was slowly added. The reaction was stirred at 10 °C for 2 hours. The product was indicated present via UPLC analysis. The mixture was filtered and the solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(6-cyanobenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide as a white solid (31.4 mg, 0.080 mmol, 23 %). 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.55 (dd, J = 40.8, 7.6 Hz, 3H), 7.86 (dd, J = 26.9, 8.5 Hz, 2H), 7.65 (d, J = 7.9 Hz, 1H), 7.37 (dd, J = 7.9, 4.8 Hz, 1H), 5.73 (s, 2H), 2.91 (t, J = 11.1 Hz, 1H), 1.66 (d, J = 8.7 Hz, 5H), 1.42 (dd, J = 23.0, 11.5 Hz, 2H), 1.32 – 1.11 (m, 3H); LCMS (ESI) m/z: 377.0 [M+H]+. Example 86. Preparation of N-(4-chloro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000128_0002
Step 1: Preparation of 4-chlorobenzo[d]thiazol-2-amine
Figure imgf000128_0003
To a solution of 4-chlorobenzo[d]thiazol-2-amine (600 mg, 3.3 mmol) and nicotinaldehyde (348 mg, 3.3 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (614 mg, 16 mmol) in ethanol (5 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 4-chlorobenzo[d]thiazol-2-amine (320 mg, 1.2 mmol, 36 %) as a yellow solid. LCMS data unavailable. Step 2: Preparation of N-(4-chloro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide
Figure imgf000129_0001
To a solution of 4-chlorobenzo[d]thiazol-2-amine (100 mg, 0.36 mmol) in dichloromethane (15 mL) was added N,N-diisopropylethylamine (139 mg, 1.1 mmol) under argon at 0 °C. The reaction was stirred at 0 °C for 10 min. Then cyclohexanecarbonyl chloride (52.6 mg, 0.36 mmol) was slowly added. The reaction was stirred at 10 °C for 2 hours. The product was indicated present via UPLC analysis. The mixture was filtered and the solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(4-chlorobenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide as a white solid (28.4 mg, 0.070 mmol, 21 %). 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.65 (d, J = 1.7 Hz, 1H), 8.50 (d, J = 4.6 Hz, 1H), 7.98 (d, J = 7.9 Hz, 1H), 7.72 (d, J = 8.0 Hz, 1H), 7.53 (d, J = 7.8 Hz, 1H), 7.42 – 7.28 (m, 2H), 5.70 (s, 2H), 2.98 (t, J = 11.2 Hz, 1H), 1.67 (t, J = 15.9 Hz, 5H), 1.54 – 1.40 (m, 2H), 1.36 – 1.12 (m, 3H); LCMS (ESI) m/z: 386.0 [M+H]+. Example 87. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(4-methylpyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000129_0002
Step 1: Preparation of 6-ethyl-N-[(4-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000129_0003
In a reaction vial, a solution of 4-methylpyridine-3-carbaldehyde (149 mg, 1.2 mmol) and 6-ethyl- 1,3-benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-ethyl-N-[(4-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (156 mg, 0.56 mmol, 49 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.51 (s, 1H), 8.42 (d, J = 5.1 Hz, 1H), 7.50 – 7.38 (m, 2H), 7.21 – 7.04 (m, 2H), 5.55 (s, 1H), 4.66 (s, 2H), 2.69 (q, J = 7.6 Hz, 2H), 2.40 (s, 3H), 1.25 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 284.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(4-methylpyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000130_0001
In a reaction vial, triethylamine (0.072 mL, 0.52 mmol) was added to a solution of 6-ethyl-N-[(4- methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (147 mg, 0.52 mmol) in dichloromethane (1 mL) and stirred at 0 °C. cyclohexanecarbonyl chloride (113 mg, 0.78 mmol) was added dropwise, and the reaction was warmed to room temperature and stir for 45 minutes. The product was indicated present via UPLC analysis. The reaction was diluted with 10 mL deionized water and the layers were separated. The inorganic layer was washed with 2x 10 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(4-methylpyridin-3-yl)methyl]cyclohexanecarboxamide (62.9 mg, 0.16 mmol, 31 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.40 (s, 1H), 8.16 (s, 1H), 7.62 (s, 2H), 7.18 (dd, J = 29.0, 6.7 Hz, 2H), 5.54 (s, 2H), 2.75 (q, J = 7.6 Hz, 2H), 2.55 (s, 1H), 2.46 (s, 3H), 1.80 (d, J = 11.7 Hz, 4H), 1.74 – 1.55 (m, 3H), 1.28 (t, J = 7.5 Hz, 4H), 1.25 – 1.05 (m, 2H); LCMS (ESI) m/z: 394.3 [M+H]+. Example 88. Preparation of N-(6-methanesulfonyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000130_0002
Step 1: Preparation of 6-(methylsulfonyl)-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000130_0003
To a solution of 6-(methylsulfonyl)benzo[d]thiazol-2-amine (600 mg, 2.6 mmol) and nicotinaldehyde (281 mg, 2.6 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (495 mg, 13 mmol) in ethanol (5 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-(methylsulfonyl)-N- (pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (340 mg, 1.1 mmol, 41 %) as a white solid. Step 2: Preparation of N-(6-methanesulfonyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000131_0001
To a solution of 6-(methylsulfonyl)-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (100 mg, 0.31 mmol) in dichloromethane (10 mL) was added N,N-diisopropylethylamine (120 mg, 0.93 mmol) under argon at 0 °C. The reaction was stirred at 0 °C for 10 min. Then cyclohexanecarbonyl chloride (67.9 mg, 0.46 mmol) was slowly added. The reaction was stirred at 10 °C for 2 hours. The product was indicated present via UPLC analysis. The mixture was filtered and the solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(6-(methylsulfonyl)benzo[d]thiazol- 2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide as a white solid (3.30 mg, 0.020 mmol, 2.5 %). 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.67 (s, 1H), 8.61 – 8.51 (m, 1H), 8.50 (d, J = 3.5 Hz, 1H), 8.06 – 7.85 (m, 2H), 7.64 (d, J = 8.0 Hz, 1H), 7.37 (dd, J = 7.9, 4.8 Hz, 1H), 5.74 (s, 2H), 3.25 (s, 3H), 2.91 (t, J = 11.1 Hz, 1H), 1.67 (d, J = 9.7 Hz, 5H), 1.43 (dd, J = 21.6, 11.0 Hz, 2H), 1.24 (d, J = 12.2 Hz, 3H); LCMS (ESI) m/z: 430.0 [M+H]+. Example 89. Preparation of ethyl 2-{N-[(pyridin-3-yl)methyl]cyclohexaneamido}-1,3-benzothiazole- 6-carboxylate
Figure imgf000131_0002
Step 1: Preparation of ethyl 2-(pyridin-3-ylmethylamino)benzo[d]thiazole-6-carboxylate
Figure imgf000131_0003
To a solution of ethyl 2-aminobenzo[d]thiazole-6-carboxylate (800 mg, 3.6 mmol) and nicotinaldehyde (385 mg, 3.6 mmol) in toluene (16 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (680 mg, 18 mmol) in ethanol (5 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer ethyl 2-(pyridin-3- ylmethylamino)benzo[d]thiazole-6-carboxylate (480 mg, 1.5 mmol, 44 %) as a yellow solid. Step 2: Preparation of ethyl 2-{N-[(pyridin-3-yl)methyl]cyclohexaneamido}-1,3-benzothiazole-6- carboxylate
Figure imgf000132_0001
To a solution of ethyl 2-(pyridin-3-ylmethylamino)benzo[d]thiazole-6-carboxylate (440 mg, 1.4 mmol) in dichloromethane (15 mL) was added N,N-diisopropylethylamine (542 mg, 4.2 mmol) under argon at 0 °C. The reaction was stirred at 0 °C for 10 min. Then cyclohexanecarbonyl chloride (307 mg, 2.1 mmol) was slowly added. The mixture was stirred at 40 °C for 17 hours. The product was indicated present via UPLC analysis. The mixture was filtered and the solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product ethyl 2-(N-(pyridin-3- ylmethyl)cyclohexanecarboxamido)benzo[d]thiazole-6-carboxylate as a white solid (180 mg, 0.42 mmol, 30 %).1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.63 (dd, J = 30.9, 1.7 Hz, 2H), 8.50 (d, J = 3.5 Hz, 1H), 7.99 (dd, J = 8.5, 1.7 Hz, 1H), 7.82 (d, J = 8.5 Hz, 1H), 7.65 (d, J = 8.0 Hz, 1H), 7.37 (dd, J = 7.9, 4.8 Hz, 1H), 5.72 (s, 2H), 4.34 (q, J = 7.1 Hz, 2H), 2.90 (t, J = 11.4 Hz, 1H), 1.67 (d, J = 9.2 Hz, 5H), 1.54 – 1.33 (m, 5H), 1.19 (d, J = 24.8 Hz, 3H); LCMS (ESI) m/z: 424.1 [M+H]+. Example 90: Preparation of N-[(pyridin-3-yl)methyl]-N-(quinolin-2-yl)cyclohexanecarboxamide
Figure imgf000132_0002
In a reaction vial, 1-(pyridin-3-yl)methanamine (989 mg, 9.2 mmol), 2-chloroquinoline (300 mg, 1.8 mmol), and N,N-diisopropylethylamine (1.59 mL, 9.2 mmol) in N-Methyl-2-pyrrolidone (9.2 mL) was heated to 120 °C and stirred overnight. The product was indicated present via UPLC analysis. The reaction was removed from heat and cooled to room temperature. The reaction was diluted with dichloromethane and washed with water (3x). The organic layers were pooled, dried over sodium sulfate, filtered and concentrated. Crude product was purified via flash chromatography eluting with 0-10% methanol in dichloromethane. Product N-[(pyridin-3-yl)methyl]quinolin-2-amine (175 mg, 0.75 mmol, 41 %) was isolated as an orange oil.1H NMR (300 MHz, Chloroform-d) δ 8.77 – 8.35 (m, 1H), 7.91 – 7.46 (m, 2H), 7.38 – 7.10 (m, 1H), 6.65 (d, J = 8.8 Hz, 1H), 5.12 (s, 1H), 4.78 (d, J = 5.8 Hz, 1H), 3.38 (t, J = 7.1 Hz, 1H), 2.85 (s, 2H), 2.38 (t, J = 8.1 Hz, 1H), 2.14 – 1.91 (m, 1H); LCMS (ESI) m/z: 236.1 [M+H]+. Step 2: Preparation of N-[(pyridin-3-yl)methyl]-N-(quinolin-2-yl)cyclohexanecarboxamide
Figure imgf000133_0001
In a reaction vial, N-[(pyridin-3-yl)methyl]quinolin-2-amine (176 mg, 0.74 mmol) and triethylamine (0.103 mL, 0.74 mmol) were dissolved in dichloromethane (3.7 mL) and cooled to 0 °C. To the solution was added cyclohexanecarbonyl chloride (0.147 mL, 1.1 mmol), slowly. The reaction was stirred from 0 °C to room temperature overnight. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane and washed with water (3x). The organic layers were pooled, dried over anhydrous sodium sulfate, filtered, and concentrated. Crude product was purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane through 24 g of silica gel. Product N-[(pyridin-3-yl)methyl]-N-(quinolin-2-yl)cyclohexanecarboxamide (112 mg, 0.33 mmol, 44 %) was afforded as an orange oil.1H NMR (300 MHz, Chloroform-d) δ 8.62 – 8.41 (m, 2H), 8.16 (d, J = 8.6 Hz, 1H), 8.10 – 7.94 (m, 1H), 7.91 – 7.68 (m, 3H), 7.58 (ddd, J = 8.1, 6.9, 1.2 Hz, 1H), 7.34 – 7.08 (m, 2H), 5.26 (d, J = 32.5 Hz, 2H), 2.50 (tt, J = 11.3, 3.5 Hz, 1H), 1.96 – 1.41 (m, 8H), 1.35 – 0.86 (m, 3H); LCMS (ESI) m/z: 346.3 [M+H]+. Example 91. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(4-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000133_0002
Step 1: Preparation of 6-ethyl-N-[(4-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000133_0003
In a reaction vial, a solution of 4-methoxypyridine-3-carbaldehyde (168 mg, 1.2 mmol) and 6- ethyl-1,3-benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-ethyl-N-[(4-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (248 mg, 0.83 mmol, 74 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.50 (s, 1H), 8.46 (d, J = 5.7 Hz, 1H), 7.50 – 7.36 (m, 2H), 7.12 (dd, J = 8.2, 1.8 Hz, 1H), 6.81 (d, J = 5.7 Hz, 1H), 5.67 (s, 1H), 4.59 (s, 2H), 3.92 (d, J = 1.6 Hz, 3H), 2.68 (q, J = 7.6 Hz, 2H), 1.24 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 300.2 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(4-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000134_0001
In a reaction vial, triethylamine (82.0 mg, 0.81 mmol) was added to a solution of 6-ethyl-N-[(4- methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (244 mg, 0.81 mmol) in dichloromethane (1 mL) and stirred at 0 °C. cyclohexanecarbonyl chloride (177 mg, 1.2 mmol) was added dropwise, and the reaction was warmed to room temperature and stir for 45 minutes. The product was indicated present via UPLC analysis. The reaction was diluted with 10 mL deionized water and the layers were separated. The inorganic layer was washed with 2x 10 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(4-methoxypyridin-3-yl)methyl]cyclohexanecarboxamide (62.5 mg, 0.15 mmol, 19 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.43 (d, J = 5.7 Hz, 1H), 8.25 (s, 1H), 7.66 (d, J = 8.3 Hz, 1H), 7.61 (s, 1H), 7.23 (dd, J = 8.3, 1.4 Hz, 1H), 6.83 (d, J = 5.8 Hz, 1H), 5.55 (s, 2H), 3.95 (s, 3H), 2.74 (p, J = 7.3 Hz, 3H), 1.80 (d, J = 12.6 Hz, 4H), 1.71 – 1.51 (m, 3H), 1.28 (t, J = 7.6 Hz, 3H), 1.26 – 1.13 (m, 3H); LCMS (ESI) m/z: 410.3 [M+H]+. Example 92. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(5-fluoropyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000134_0002
Step 1: Preparation of 6-ethyl-N-[(5-fluoropyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000134_0003
In a reaction vial, a solution of 5-fluoropyridine-3-carbaldehyde (153 mg, 1.2 mmol) and 6-ethyl- 1,3-benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-ethyl-N-[(5-fluoropyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (320 mg, 1.1 mmol, 99 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.47 (d, J = 1.6 Hz, 1H), 8.41 (d, J = 2.7 Hz, 1H), 7.51 (ddd, J = 9.1, 2.7, 1.8 Hz, 1H), 7.48 – 7.39 (m, 2H), 7.15 (dd, J = 8.2, 1.8 Hz, 1H), 5.70 (s, 1H), 4.73 (s, 2H), 2.69 (q, J = 7.6 Hz, 2H), 1.25 (td, J = 7.6, 0.8 Hz, 3H); LCMS (ESI) m/z: 288.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(5-fluoropyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000135_0001
In a reaction vial, triethylamine (0.18 mL, 1.3 mmol) was added to a solution of 6-ethyl-N-[(5- fluoropyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (321 mg, 1.1 mmol) in dichloromethane (1.1 mL) and stirred at 0 °C. cyclohexanecarbonyl chloride (243 mg, 1.7 mmol) was added dropwise, and the reaction was warmed to room temperature and stir for 45 minutes. The product was indicated present via UPLC. The reaction was diluted with 10 mL deionized water and the layers were separated. The inorganic layer was washed with 2x 10 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-50% ethyl acetate in hexanes. Product N-(6-ethyl-1,3- benzothiazol-2-yl)-N-[(5-fluoropyridin-3-yl)methyl]cyclohexanecarboxamide (308 mg, 0.78 mmol, 70 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.43 (s, 1H), 8.40 (d, J = 2.7 Hz, 1H), 7.68 (d, J = 8.3 Hz, 1H), 7.63 (s, 1H), 7.37 – 7.25 (m, 1H), 5.60 (s, 2H), 2.77 (q, J = 7.6 Hz, 2H), 2.70 – 2.60 (m, 1H), 1.70 (ddd, J = 33.7, 25.6, 12.4 Hz, 6H), 1.29 (t, J = 7.6 Hz, 3H), 1.29 – 1.16 (m, 3H); LCMS (ESI) m/z: 398.3 [M+H]+. Example 93. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000135_0002
Step 1: Preparation of 6-ethyl-N-[(1H-pyrazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000135_0003
In a reaction vial, a solution of 1H-pyrazole-5-carbaldehyde (118 mg, 1.2 mmol) and 6-ethyl-1,3- benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-ethyl-N-[(1H- pyrazol-5-yl)methyl]-1,3-benzothiazol-2-amine (247 mg, 0.96 mmol, 85 %) was afforded as a cream solid. 1H NMR (300 MHz, Methanol-d4) δ 7.58 (s, 1H), 7.44 – 7.29 (m, 2H), 7.09 (d, J = 8.0 Hz, 1H), 6.33 (s, 1H), 4.62 (s, 2H), 2.73 – 2.51 (m, 2H), 1.22 (tt, J = 7.5, 1.4 Hz, 3H); LCMS (ESI) m/z: 259.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000136_0001
In a reaction vial, triethylamine (0.16 mL, 1.1 mmol) was added to a solution of 6-ethyl-N-[(1H- pyrazol-5-yl)methyl]-1,3-benzothiazol-2-amine (248 mg, 0.96 mmol) in dichloromethane (1 mL) and stirred at 0 °C. cyclohexanecarbonyl chloride (209 mg, 1.4 mmol) was added dropwise, and the reaction was warmed to room temperature and stir for 2 hours. The product was confirmed present via UPLC analysis. The reaction was diluted with 10 mL deionized water and the layers were separated. The inorganic layer was washed with 2x 10 mL portions of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-50% ethyl acetate in hexanes. N-(6-ethyl-1,3- benzothiazol-2-yl)-N-[(1H-pyrazol-5-yl)methyl]cyclohexanecarboxamide (206 mg, 0.56 mmol, 59 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.19 (d, J = 2.8 Hz, 1H), 7.50 (d, J = 8.2 Hz, 1H), 7.47 – 7.40 (m, 1H), 7.16 (dd, J = 8.2, 1.8 Hz, 1H), 6.44 (d, J = 2.8 Hz, 1H), 5.74 (s, 1H), 4.71 (s, 2H), 3.61 (tt, J = 11.5, 3.5 Hz, 1H), 2.70 (q, J = 7.6 Hz, 2H), 1.98 (dd, J = 12.5, 2.5 Hz, 2H), 1.90 – 1.68 (m, 3H), 1.66 – 1.30 (m, 5H), 1.26 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 369.3 [M+H]+. Example 94. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]bicyclo[1.1.1]pentane-1-carboxamide
Figure imgf000136_0002
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]bicyclo[1.1.1]pentane-1- carboxamide
Figure imgf000136_0003
A solution of bicyclo[1.1.1]pentane-1-carboxylic acid (67.0 mg, 0.60 mmol) in thionyl chloride (2 mL) was stirred at 70 °C for 2 hours. The reaction was concentrated under reduced pressure and re- dissolved in dichloromethane (2 mL).6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (80.0 mg, 0.30 mmol) and N,N-diisopropylethylamine (77.0 mg, 0.60 mmol) was added to the solution at 0 °C. The reaction was warmed to room temperature with stirring over 16 hours. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane (20 mL) and washed with water (20 mL). The organic layer separated and concentrated under reduced pressure. Crude product was purified by Prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) obtaining N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)bicyclo[1.1.1]pentane-1-carboxamide (24.0 mg, 0.070 mmol, 22 %) as white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.52 – 8.38 (m, 2H), 7.82 (s, 1H), 7.60 (d, J = 8.3 Hz, 1H), 7.54 (d, J = 7.9 Hz, 1H), 7.35 (dd, J = 7.8, 4.8 Hz, 1H), 7.25 (dd, J = 8.4, 1.6 Hz, 1H), 5.64 (s, 2H), 2.70 (q, J = 7.5 Hz, 2H), 2.44 (s, 1H), 2.13 (s, 6H), 1.21 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 364.2 [M+H]+. Example 95. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]oxane-4- carboxamide
Figure imgf000137_0001
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]oxane-4-carboxamide
Figure imgf000137_0002
In a reaction vial, tetrahydro-2H-pyran-4-carbonyl chloride (89.0 mg, 0.60 mmol) was added into a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (80.0 mg, 0.30 mmol) and N,N- diisopropylethylamine (77.0 mg, 0.60 mmol) in dichloromethane (2 mL) at 0 °C. The reaction was warmed to room temperature over 16 hours. The product was indicated present via UPLC analysis. The mixture was diluted with dichloromethane (20 mL) and washed with water (20 mL). The organic layer was concentrated under reduced pressure. Crude product was purified by Prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) obtaining N-(6-ethylbenzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)tetrahydro-2H-pyran-4-carboxamide (62.0 mg, 0.16 mmol, 54 %) as white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.57 (d, J = 1.9 Hz, 1H), 8.48 (dd, J = 4.8, 1.4 Hz, 1H), 7.82 (d, J = 1.1 Hz, 1H), 7.64 (t, J = 8.3 Hz, 2H), 7.36 (dd, J = 7.9, 4.8 Hz, 1H), 7.28 (dd, J = 8.3, 1.6 Hz, 1H), 5.70 (s, 2H), 3.83 (dd, J = 11.5, 2.5 Hz, 2H), 3.34 (m, 2H), 3.26 – 3.13 (m, 1H), 2.71 (q, J = 7.6 Hz, 2H), 1.77 – 1.61 (m, 2H), 1.56 (d, J = 11.9 Hz, 2H), 1.22 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 382.2 [M+H]+. Example 96. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-4-phenyl-N-[(pyridin-3- yl)methyl]butanamide
Figure imgf000137_0003
Step 1: Preparation of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000138_0001
To a solution of nicotinaldehyde (481 mg, 4.5 mmol), 5-fluorobenzo[d]thiazol-2-amine (500 mg, 3.0 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (342 mg, 9.0 mmol) in ethanol (10 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (600 mg, 2.3 mmol, 77 %) as a white solid. LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-4-phenyl-N-[(pyridin-3-yl)methyl]butanamide
Figure imgf000138_0002
To a solution of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (100 mg, 0.39 mmol) in N,N-dimethylformamide (5 mL) was added 4-phenylbutanoic acid (127 mg, 0.77 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (292 mg, 0.77 mmol) and N,N-diisopropylethylamine (115 mg, 0.77 mmol). The reaction mixture was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. The mixture was purified prep-HPLC to give N-(5-fluoro-1,3-benzothiazol-2-yl)-4-phenyl- N-[(pyridin-3-yl)methyl]butanamide (22.5 mg, 0.056 mmol, 28 %) as white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.57 – 8.45 (m, 2H), 8.04 (dd, J = 8.8, 5.5 Hz, 1H), 7.59 (dd, J = 10.1, 2.6 Hz, 2H), 7.43 – 7.25 (m, 1H), 7.25 – 7.07 (m, 6H), 5.59 (s, 2H), 2.83 – 2.52 (m, 4H), 2.13 – 1.63 (m, 2H); LCMS (ESI) m/z: 406.0 [M+H]+. Example 97. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000138_0003
Step 1: Preparation of 6-fluoro-N-[(1H-pyrazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000138_0004
In a reaction vial, a solution of 1H-pyrazole-5-carbaldehyde (123 mg, 1.3 mmol) and 6-fluoro-1,3- benzothiazol-2-amine (200 mg, 1.2 mmol) in dry toluene (12 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry ethyl alcohol (12 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-fluoro-N-[(1H- pyrazol-5-yl)methyl]-1,3-benzothiazol-2-amine (204 mg, 0.82 mmol, 70 %) was afforded as a yellow solid. 1H NMR (300 MHz, Methanol-d4) δ 7.59 (s, 1H), 7.38 (s, 2H), 7.00 (t, J = 9.3 Hz, 1H), 6.33 (s, 1H), 4.62 (s, 2H); LCMS (ESI) m/z: 249.1 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000139_0001
In a reaction vial, triethylamine (99.6 mg, 0.99 mmol) was added to a solution of 6-fluoro-N-[(1H- pyrazol-5-yl)methyl]-1,3-benzothiazol-2-amine (204 mg, 0.82 mmol) in dichloromethane (1 mL) and stirred at 0 °C. cyclohexanecarbonyl chloride (180 mg, 1.2 mmol) was added dropwise, and the reaction was warmed to room temperature and stir for 1.5 hours. The product was indicated present via UPLC analysis. The reaction was diluted with 10 mL deionized water and the layers were separated. The inorganic layer was washed with 2x 10 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-50% ethyl acetate in hexanes. Product N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-5-yl)methyl]cyclohexanecarboxamide (221 mg, 0.62 mmol, 75 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.21 (d, J = 2.8 Hz, 1H), 7.51 (dd, J = 8.9, 4.7 Hz, 1H), 7.32 (dd, J = 8.1, 2.6 Hz, 1H), 7.07 (td, J = 8.9, 2.7 Hz, 1H), 6.46 (d, J = 2.8 Hz, 1H), 4.70 (s, 2H), 3.60 (tt, J = 11.5, 3.4 Hz, 1H), 1.97 (dd, J = 12.5, 2.6 Hz, 1H), 1.88 – 1.71 (m, 2H), 1.64 – 1.47 (m, 2H), 1.47 – 1.19 (m, 1H); LCMS (ESI) m/z: 359.2 [M+H]+. Example 98. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-4-methyl-N-[(pyridin-3- yl)methyl]piperazine-1-carboxamide
Figure imgf000139_0002
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000140_0001
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-4-methyl-N-[(pyridin-3-yl)methyl]piperazine-1- carboxamide
Figure imgf000140_0002
To a solution of triphosgene (65.3 mg, 0.22 mmol) in dichloromethane (10 mL) was added a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (120 mg, 0.45 mmol) and pyridine (142 mg, 1.8 mmol) in dichloromethane (5 mL) at -60 °C under argon. The reaction was warmed to room temperature over 2 hours. Then a solution of 1-methylpiperazine (67.0 mg, 0.67 mmol) and pyridine (142 mg, 1.8 mmol) in dichloromethane (6 mL) was added at -60 °C. The resulting mixture was warmed to room temperature over 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(6- ethylbenzo[d]thiazol-2-yl)-4-methyl-N-(pyridin-3-ylmethyl)piperazine-1-carboxamide as a white solid (17.6 mg, 0.040 mmol, 9.9 %).1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.58 (s, 1H), 8.45 (d, J = 4.7 Hz, 1H), 8.28 (s, 1H), 7.76 (d, J = 7.9 Hz, 1H), 7.70 (s, 1H), 7.56 (d, J = 8.3 Hz, 1H), 7.35 (dd, J = 7.9, 4.8 Hz, 1H), 7.22 (d, J = 8.3 Hz, 1H), 5.12 (s, 2H), 3.39 (d, J = 4.3 Hz, 4H), 2.66 (q, J = 7.5 Hz, 2H), 2.28 (s, 4H), 2.15 (s, 3H), 1.19 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 396.1 [M+H]+. Example 99. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclopentanecarboxamide
Figure imgf000140_0003
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000141_0001
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclopentanecarboxamide
Figure imgf000141_0002
To a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (120 mg, 0.45 mmol) in dichloromethane (10 mL) was added N,N-diisopropylethylamine (174 mg, 1.4 mmol) under argon at 0 °C. The solution was stirred at 0 °C for 10 min, then cyclopentanecarbonyl chloride (88.8 mg, 0.67 mmol) was slowly added. The reaction was warmed to room temperature over 2 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclopentanecarboxamide as a white solid (41.5 mg, 0.11 mmol, 25 %).1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.53 (d, J = 1.9 Hz, 1H), 8.49 – 8.41 (m, 1H), 7.81 (s, 1H), 7.61 (dd, J = 16.8, 8.1 Hz, 2H), 7.36 (d, J = 4.7 Hz, 1H), 7.27 (dd, J = 8.3, 1.5 Hz, 1H), 5.67 (s, 2H), 3.26 (dd, J = 15.2, 7.7 Hz, 1H), 2.71 (q, J = 7.6 Hz, 2H), 1.85 – 1.65 (m, 6H), 1.62 – 1.51 (m, 2H), 1.22 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 366.1 [M+H]+. Example 100. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclobutanecarboxamide
Figure imgf000141_0003
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclobutanecarboxamide
Figure imgf000142_0001
To a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (80.0 mg, 0.30 mmol) and N,N-diisopropylethylamine (80.0 mg, 0.60 mmol) in dichloromethane (4 mL) was added cyclobutanecarbonyl chloride (70.0 mg, 0.60 mmol) at 0 °C under argon. The reaction was warmed to room temperature with stirring over 16 hours. The product was indicated present via UPLC analysis. Water (30 mL) was added, then the mixture was extracted with dichloromethane (2x 30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclobutanecarboxamide (40.0 mg, 0.11 mmol, 38 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.51 (d, J = 1.9 Hz, 1H), 8.46 (dd, J = 4.7, 1.3 Hz, 1H), 7.82 (s, 1H), 7.62 (d, J = 8.3 Hz, 1H), 7.57 (d, J = 7.9 Hz, 1H), 7.33 (dd, J = 7.8, 4.8 Hz, 1H), 7.26 (dd, J = 8.3, 1.6 Hz, 1H), 5.48 (s, 2H), 3.66 (p, J = 8.3 Hz, 1H), 2.71 (q, J = 7.5 Hz, 2H), 2.35 – 2.18 (m, 2H), 2.08 (q, J = 8.9 Hz, 2H), 1.92 (dq, J = 17.6, 8.9 Hz, 1H), 1.77 (dd, J = 19.4, 9.5 Hz, 1H), 1.22 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 352.2 [M+H]+. Example 101. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-2-methyl-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000142_0002
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-2-methyl-N-[(pyridin-3-yl)methyl]propanamide
Figure imgf000142_0003
Isobutyryl chloride (64.0 mg, 0.60 mmol) was added into a solution of 6-ethyl-N-(pyridin-3- ylmethyl)benzo[d]thiazol-2-amine (80.0 mg, 0.30 mmol) and N,N-diisopropylethylamine (77.0 mg, 0.60 mmol) in dichloromethane (2 mL) at 0 °C. The reaction was warmed to room temperature with stirring over 16 hours. The product was indicated present via UPLC analysis. The mixture was diluted with dichloromethane (20 mL) and washed with water (20 mL). The organic layer was separated and concentrated under reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to afford N-(6-ethylbenzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)isobutyramide (37.0 mg, 0.10 mmol, 32 %) as white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.54 (d, J = 1.8 Hz, 1H), 8.48 (dd, J = 4.7, 1.4 Hz, 1H), 7.82 (s, 1H), 7.65 (d, J = 8.3 Hz, 1H), 7.60 (d, J = 8.0 Hz, 1H), 7.36 (dd, J = 7.9, 4.9 Hz, 1H), 7.28 (dd, J = 8.3, 1.6 Hz, 1H), 5.67 (s, 2H), 3.12 (dt, J = 13.3, 6.7 Hz, 1H), 2.71 (q, J = 7.5 Hz, 2H), 1.22 (t, J = 7.6 Hz, 3H), 1.09 (d, J = 6.6 Hz, 6H); LCMS (ESI) m/z: 340.2 [M+H]+. Example 102. Preparation of N-(5,6-dimethoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000143_0001
Step 1: Preparation of 5,6-dimethoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000143_0002
To a solution of nicotinaldehyde (0.890 g, 8.4 mmol), 5,6-dimethoxybenzo[d]thiazol-2-amine (1.20 g, 5.6 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (0.638 g, 17 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 5,6-dimethoxy-N-(pyridin-3- ylmethyl)benzo[d]thiazol-2-amine (1.00 g, 3.3 mmol, 59 %) as a white solid. LCMS (ESI) m/z: 302.1 [M+H]+. Step 2: Preparation of N-(5,6-dimethoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000143_0003
To a solution of 5,6-dimethoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (300 mg, 1.0 mmol), cyclohexanecarboxylic acid (128 mg, 1.0 mmol), triethylamine (303 mg, 3.0 mmol) in dichloromethane (10 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (570 mg, 1.5 mmol). The mixture was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%- 95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to offer N-(5,6-dimethoxybenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide (40.0 mg, 0.090 mmol, 23 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.55 (d, J = 2.0 Hz, 1H), 8.49 (dd, J = 5.0, 1.5 Hz, 1H), 7.61 (d, J = 8.0 Hz, 1H), 7.55 (s, 1H), 7.37 (dd, J = 8.0, 5.0 Hz, 1H), 7.31 (s, 1H), 5.65 (s, 2H), 3.79 (s, 3H), 3.80 (s, 3H), 2.84 – 2.80 (m, 1H), 1.68 – 1.62 (m, 5H), 1.45 – 1.41 (m, 2H), 1.24 – 1.21 (m, 3H); LCMS (ESI) m/z: 412.2 [M+H]+. Example 103. Preparation of 2-(benzyloxy)-N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]acetamide
Figure imgf000144_0001
Step 1: Preparation of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000144_0002
To a solution of nicotinaldehyde (481 mg, 4.5 mmol), 5-fluorobenzo[d]thiazol-2-amine (500 mg, 3.0 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (342 mg, 9.0 mmol) in methanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (600 mg, 2.3 mmol, 77 %) as a white solid. LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of 2-(benzyloxy)-N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]acetamide
Figure imgf000144_0003
To a solution of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (200 mg, 0.77 mmol) in N,N-dimethylformamide (10 mL) was added 2-(benzyloxy)acetic acid (256 mg, 1.5 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (585 mg, 1.5 mmol) and N,N-diisopropylethylamine (198 mg, 1.5 mmol). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. Water (20 mL) was added to reaction mixture, which was then extracted with 2 x 20 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified prep-HPLC to give 2-(benzyloxy)-N-(5-fluoro-1,3-benzothiazol-2-yl)-N- [(pyridin-3-yl)methyl]acetamide (75.0 mg, 0.18 mmol, 36 %) as white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.54 (d, J = 1.9 Hz, 1H), 8.48 (dd, J = 4.7, 1.3 Hz, 1H), 8.07 (dd, J = 8.8, 5.5 Hz, 1H), 7.67 – 7.56 (m, 2H), 7.42 – 7.20 (m, 7H), 5.51 (s, 2H), 4.58 (d, J = 2.9 Hz, 4H); LCMS (ESI) m/z: 408.1 [M+H]+. Example 104. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-1,1-dioxo-N-[(pyridin-3-yl)methyl]- 1λ⁶-thiomorpholine-4-carboxamide
Figure imgf000145_0001
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000145_0002
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-1,1-dioxo-N-[(pyridin-3-yl)methyl]-1λ⁶- thiomorpholine-4-carboxamide
Figure imgf000145_0003
To a solution of triphosgene (65.3 mg, 0.22 mmol) in dichloromethane (10 mL) was added a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (120 mg, 0.45 mmol) and pyridine (142 mg, 1.8 mmol) in dichloromethane (5 mL) at -60 °C under argon. The reaction was warmed to room temperature over 2 hours. Then a solution of thiomorpholine 1,1-dioxide (87.8 mg, 0.67 mmol) and pyridine (142 mg, 1.8 mmol) in dichloromethane (6 mL) was added at -60 °C. The resulting mixture was warmed to room temperature over 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(6- ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)thiomorpholine-4-carboxamide 1,1-dioxide (43.6 mg, 0.10 mmol, 23 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.59 (d, J = 1.7 Hz, 1H), 8.45 (t, J = 11.3 Hz, 1H), 7.82 – 7.67 (m, 2H), 7.58 (d, J = 8.3 Hz, 1H), 7.33 (dd, J = 7.8, 4.8 Hz, 1H), 7.28 – 7.18 (m, 1H), 5.15 (s, 2H), 3.81 (s, 4H), 3.26 (s, 4H), 2.66 (q, J = 7.5 Hz, 2H), 1.29 – 1.07 (m, 3H); LCMS (ESI) m/z: 431.0 [M+H]+. Example 105. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]oxane-3- carboxamide
Figure imgf000146_0001
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000146_0002
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]oxane-3-carboxamide
Figure imgf000146_0003
To a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (120 mg, 0.45 mmol) in dichloromethane (15 mL) were added tetrahydro-2H-pyran-3-carboxylic acid (87.1 mg, 0.67 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (255 mg, 0.67 mmol) and triethylamine (136 mg, 1.4 mmol) at 0 °C. The reaction was warmed to room temperature with stirring over 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C18 21*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product N-(6- ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)tetrahydro-2H-pyran-3-carboxamide (75.9 mg, 0.19 mmol, 44 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.56 (d, J = 1.9 Hz, 1H), 8.50 (dd, J = 4.7, 1.2 Hz, 1H), 7.82 (s, 1H), 7.64 (dd, J = 11.9, 8.2 Hz, 2H), 7.37 (dd, J = 7.9, 4.8 Hz, 1H), 7.28 (dd, J = 8.3, 1.5 Hz, 1H), 5.82 – 5.57 (m, 2H), 3.83 (dd, J = 25.5, 11.0 Hz, 2H), 3.45 (t, J = 10.6 Hz, 1H), 3.32 (td, J = 11.1, 5.0 Hz, 1H), 3.09 (dd, J = 12.3, 8.7 Hz, 1H), 2.71 (q, J = 7.6 Hz, 2H), 1.82 (d, J = 12.3 Hz, 1H), 1.68 (ddd, J = 17.4, 11.6, 5.7 Hz, 1H), 1.54 (dd, J = 11.1, 8.6 Hz, 2H), 1.22 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 382.2 [M+H]+. Example 106. Preparation of 4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]benzamide
Figure imgf000147_0001
Step 1: Preparation of 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000147_0002
To a solution of 1H-imidazole-4-carbaldehyde (2.50 g, 27 mmol), 6-fluorobenzo[d]thiazol-2-amine (3.00 g, 18 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.30 g, 36 mmol) in methanol (50 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3- benzothiazol-2-amine (2.50 g, 10 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 249.1 [M+H]+. Step 2: Preparation of 4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]benzamide
Figure imgf000147_0003
To a solution of N-((1H-imidazol-4-yl)methyl)-6-fluorobenzo[d]thiazol-2-amine (200 mg, 0.81 mmol) in N,N-dimethylformamide (10 mL) was added 4-cyanobenzoic acid (236 mg, 1.6 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (611 mg, 1.6 mmol) and N,N-diisopropylethylamine (207 mg, 1.6 mmol). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. Water (20 mL) was added and the reaction was extracted with 2x 20 mL washes of ethyl acetate. The organic layers were pooled and concentrated under reduced pressure. The mixture was purified prep-HPLC to give 4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]benzamide (62.0 mg, 0.16 mmol, 23 %) as white solid.1H NMR (400 MHz, Dimethylsulfoxide- d6) δ 12.02 – 11.97 (S, 1H), 8.35 (s, 1H), 8.05 – 7.90 (m, 5H), 7.84 (dt, J = 23.0, 11.5 Hz, 1H), 7.52 (d, J = 1.1 Hz, 3H), 7.32 (td, J = 9.1, 2.7 Hz, 1H), 6.95 (s, 1H), 5.23 (s, 2H); LCMS (ESI) m/z: 378.1 [M+H]+. Example 107. Preparation of tert-butyl 4-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000147_0004
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000148_0001
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of tert-butyl 4-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000148_0002
To a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (300 mg, 1.1 mmol) in dichloromethane (15 mL) were added 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (380 mg, 1.7 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (631 mg, 1.7 mmol) and triethylamine (336 mg, 3.3 mmol) at 0 °C. The reaction was warmed to room temperature with stirring for 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C18 21*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product tert- butyl 4-((6-ethylbenzo[d]thiazol-2-yl)(pyridin-3-ylmethyl)carbamoyl)piperidine-1-carboxylate (167 mg, 0.35 mmol, 31 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.58 (d, J = 1.9 Hz, 1H), 8.49 (dd, J = 4.7, 1.4 Hz, 1H), 7.82 (d, J = 1.0 Hz, 1H), 7.64 (t, J = 7.1 Hz, 2H), 7.36 (dd, J = 7.8, 4.8 Hz, 1H), 7.28 (dd, J = 8.3, 1.5 Hz, 1H), 5.69 (s, 2H), 3.93 (d, J = 10.6 Hz, 2H), 3.14 (t, J = 11.1 Hz, 1H), 2.71 (dd, J = 15.1, 7.6 Hz, 4H), 1.65 (d, J = 12.0 Hz, 2H), 1.56 – 1.46 (m, 2H), 1.40 (s, 9H), 1.22 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 481.3 [M+H]+. Example 108. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000148_0003
Step 1: Preparation of 6-fluoro-N-[(1H-pyrazol-4-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000149_0001
In a reaction vial, a solution of 1H-pyrazole-4-carbaldehyde (113 mg, 1.2 mmol) and 6-fluoro-1,3- benzothiazol-2-amine (200 mg, 1.2 mmol) in dry toluene (12 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry ethyl alcohol (12 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-fluoro-N-[(1H- pyrazol-4-yl)methyl]-1,3-benzothiazol-2-amine (220 mg, 0.89 mmol, 75 %) was afforded as a white solid. 1H NMR (300 MHz, Methanol-d4) δ 7.65 (s, 2H), 7.39 (tdd, J = 8.6, 4.7, 1.9 Hz, 2H), 7.02 (tt, J = 9.0, 2.2 Hz, 1H), 4.52 (d, J = 1.7 Hz, 2H); LCMS (ESI) m/z: 249.0 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000149_0002
In a reaction vial, 6-fluoro-N-[(1H-pyrazol-4-yl)methyl]-1,3-benzothiazol-2-amine (211 mg, 0.85 mmol) and triethylamine (0.17 mL, 1.3 mmol) were dissolved in tetrahydrofuran (1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (247 mg, 1.7 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of dichloromethane were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-50% ethyl acetate in hexanes. Product N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-4- yl)methyl]cyclohexanecarboxamide (59.0 mg, 0.16 mmol, 20 %) was afforded as a white solid. 1H NMR (300 MHz, Chloroform-d) δ 8.25 (t, J = 0.8 Hz, 1H), 7.76 (d, J = 0.8 Hz, 1H), 7.49 (dd, J = 8.9, 4.7 Hz, 1H), 7.30 (dd, J = 8.1, 2.6 Hz, 1H), 7.04 (td, J = 9.0, 2.6 Hz, 1H), 5.39 (s, 1H), 4.56 (s, 2H), 3.60 (tt, J = 11.6, 3.5 Hz, 1H), 2.03 – 1.92 (m, 2H), 1.88 – 1.78 (m, 2H), 1.74 (d, J = 12.7 Hz, 1H), 1.65 – 1.43 (m, 3H), 1.46 – 1.20 (m, 3H); LCMS (ESI) m/z: 359.2 [M+H]+. Example 109. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000149_0003
Step 1: Preparation of 6-ethyl-N-[(1H-pyrazol-4-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000150_0001
In a reaction vial, a solution of 1H-pyrazole-4-carbaldehyde (107 mg, 1.1 mmol) and 6-ethyl-1,3- benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-ethyl-N-[(1H- pyrazol-4-yl)methyl]-1,3-benzothiazol-2-amine (221 mg, 0.86 mmol, 76 %) was afforded as a white solid. 1H NMR (300 MHz, Methanol-d4) δ 7.65 (s, 2H), 7.42 (d, J = 1.9 Hz, 1H), 7.36 (dd, J = 8.2, 1.9 Hz, 1H), 7.11 (dd, J = 8.2, 2.0 Hz, 1H), 4.52 (d, J = 1.9 Hz, 2H), 2.67 (q, J = 7.7 Hz, 2H), 1.24 (td, J = 7.6, 2.0 Hz, 3H); LCMS (ESI) m/z: 259.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000150_0002
In a reaction vial, 6-ethyl-N-[(1H-pyrazol-4-yl)methyl]-1,3-benzothiazol-2-amine (217 mg, 0.84 mmol) and triethylamine (0.17 mL, 1.3 mmol) were dissolved in tetrahydrofuran (1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (244 mg, 1.7 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of dichloromethane were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-50% ethyl acetate in hexanes. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-pyrazol-4- yl)methyl]cyclohexanecarboxamide (54.7 mg, 0.15 mmol, 18 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.24 (q, J = 0.8 Hz, 1H), 7.76 (d, J = 0.9 Hz, 1H), 7.48 (d, J = 8.3 Hz, 1H), 7.44 – 7.41 (m, 1H), 7.15 (dd, J = 8.2, 1.7 Hz, 1H), 5.35 (s, 1H), 4.56 (s, 2H), 3.59 (tt, J = 11.5, 3.5 Hz, 1H), 2.69 (q, J = 7.6 Hz, 2H), 2.03 – 1.91 (m, 2H), 1.89 – 1.78 (m, 2H), 1.79 – 1.67 (m, 1H), 1.63 – 1.29 (m, 5H), 1.26 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 369.3 [M+H]+. Example 110. Preparation of N-[(pyridin-3-yl)methyl]-N-(4,5,6,7-tetrahydro-1,3-benzothiazol-2- yl)cyclohexanecarboxamide
Figure imgf000151_0001
Step 1: Preparation of 4,5,6,7-tetrahydro-1,3-benzothiazol-2-amine
Figure imgf000151_0002
In a reaction vial, a solution of pyridine-3-carbaldehyde (138 mg, 1.3 mmol) and 4,5,6,7- tetrahydro-1,3-benzothiazol-2-amine (200 mg, 1.3 mmol) in dry toluene (13 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (111 mg, 3.0 mmol) in dry ethyl alcohol (13 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product N-[(pyridin-3-yl)methyl]-4,5,6,7-tetrahydro-1,3-benzothiazol-2-amine (264 mg, 1.1 mmol, 84 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.61 (d, J = 2.3 Hz, 1H), 8.54 (dd, J = 4.8, 1.7 Hz, 1H), 7.72 (dt, J = 7.8, 2.0 Hz, 1H), 7.27 (dd, J = 7.8, 4.8 Hz, 1H), 4.48 (s, 2H), 2.54 (tt, J = 3.8, 2.1 Hz, 4H), 1.79 (p, J = 3.6 Hz, 4H); LCMS (ESI) m/z: 246.2 [M+H]+. Step 2: Preparation of N-[(pyridin-3-yl)methyl]-N-(4,5,6,7-tetrahydro-1,3-benzothiazol-2- yl)cyclohexanecarboxamide
Figure imgf000151_0003
In a reaction vial, N-[(pyridin-3-yl)methyl]-4,5,6,7-tetrahydro-1,3-benzothiazol-2-amine (256 mg, 1.0 mmol) and triethylamine (0.21 mL, 1.5 mmol) were dissolved in tetrahydrofuran (1.03 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (302 mg, 2.1 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of dichloromethane were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product N-[(pyridin-3-yl)methyl]-N-(4,5,6,7-tetrahydro-1,3-benzothiazol-2- yl)cyclohexanecarboxamide (271 mg, 0.76 mmol, 74 %) was afforded as an orange waxy oil. 1H NMR (300 MHz, Chloroform-d) δ 8.55 (dd, J = 7.6, 2.2 Hz, 2H), 7.70 – 7.52 (m, 1H), 7.34 (s, 1H), 5.71 – 5.39 (m, 2H), 2.80 – 2.43 (m, 5H), 1.91 – 1.47 (m, 12H), 1.16 (d, J = 18.7 Hz, 4H); LCMS (ESI) m/z: 356.2 [M+H]+. Example 111. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperidine-1- carboxamide
Figure imgf000152_0001
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000152_0002
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperidine-1-carboxamide
Figure imgf000152_0003
To a solution of triphosgene (65.3 mg, 0.22 mmol) in dichloromethane (10 mL) was added a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (120 mg, 0.45 mmol) and pyridine (142 mg, 1.8 mmol) in dichloromethane (5 mL) at -60 °C under argon. The mixture was warmed to 0 °C and stirred for 30 minutes. Then a solution of piperidine (56.9 mg, 0.67 mmol) and pyridine (142 mg, 1.8 mmol) in dichloromethane (6 mL) was added at -60 °C. The reaction was warmed to room temperature over 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)piperidine-1-carboxamide (31.3 mg, 0.080 mmol, 18 %) was afforded as a yellow solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.59 (d, J = 1.7 Hz, 1H), 8.46 (dd, J = 4.7, 1.3 Hz, 1H), 7.77 (d, J = 7.9 Hz, 1H), 7.68 (d, J = 1.2 Hz, 1H), 7.56 (d, J = 8.2 Hz, 1H), 7.35 (dd, J = 7.8, 4.8 Hz, 1H), 7.21 (dd, J = 8.3, 1.6 Hz, 1H), 5.12 (s, 2H), 3.38 – 3.28 (m, 4H), 2.66 (q, J = 7.6 Hz, 2H), 1.50 (dd, J = 33.2, 4.0 Hz, 6H), 1.20 (q, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 381.0 [M+H]+. Example 112. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]morpholine-4- carboxamide
Figure imgf000153_0001
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000153_0002
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]morpholine-4-carboxamide
Figure imgf000153_0003
To a solution of triphosgene (65.3 mg, 0.20 mmol) in dichloromethane (10 mL) was added a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (120 mg, 0.45 mmol) and pyridine (142 mg, 1.8 mmol) in dichloromethane (5 mL) at -60 °C under argon. The mixture was warmed to 0 °C and stirred for 30 minutes. Then a solution of morpholine (58.3 mg, 0.67 mmol) and pyridine (142 mg, 1.8 mmol) in dichloromethane (6 mL) was added at -60 °C. The reaction was warmed to room temperature over 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)morpholine-4-carboxamide (19.4 mg, 0.050 mmol, 11 %) was afforded as a yellow solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.60 (s, 1H), 8.42 (t, J = 21.4 Hz, 1H), 7.77 (d, J = 7.9 Hz, 1H), 7.69 (d, J = 1.0 Hz, 1H), 7.57 (d, J = 8.2 Hz, 1H), 7.34 (dd, J = 7.8, 4.8 Hz, 1H), 7.21 (dd, J = 8.3, 1.5 Hz, 1H), 5.13 (s, 2H), 3.63 – 3.55 (m, 4H), 3.42 – 3.36 (m, 4H), 2.65 (q, J = 7.5 Hz, 2H), 1.19 (q, J = 7.7 Hz, 3H); LCMS (ESI) m/z: 383.0 [M+H]+. Example 113. Preparation of N-(6-ethoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000153_0004
Step 1: Preparation of 6-ethoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000154_0001
To a solution of nicotinaldehyde (1.07 g, 10 mmol), 6-ethoxybenzo[d]thiazol-2-amine (1.94 g, 10 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.90 g, 50 mmol) in ethanol (100 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-80% ethyl acetate in hexanes to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (0.430 g, 1.5 mmol, 15 %) as a white powder; LCMS (ESI) m/z: 286.2 [M+H]+. Step 2: Preparation of N-(6-ethoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000154_0002
Into a solution of 6-ethoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (200 mg, 0.70 mmol) and N,N-diisopropylethylamine (180 mg, 1.4 mmol) in dichloromethane (5 mL) was added cyclohexanecarbonyl chloride (210 mg, 1.4 mmol) at 0 °C. The reaction was warmed to room temperature with stirring over 4 hours. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane (20 mL) and washed with water (20 mL). The organic layer was concentrated under reduced pressure and purified by Prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product N-(6-ethoxybenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide (140 mg, 0.35 mmol, 51 %) was afforded as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.55 (d, J = 1.9 Hz, 1H), 8.51 – 8.45 (m, 1H), 7.62 (m, 2H), 7.56 (d, J = 2.5 Hz, 1H), 7.37 (dd, J = 7.9, 4.8 Hz, 1H), 7.00 (dd, J = 8.8, 2.5 Hz, 1H), 5.65 (s, 2H), 4.07 (q, J = 6.9 Hz, 2H), 2.84 (t, J = 11.3 Hz, 1H), 1.64 (m, 5H), 1.48 – 1.11 (m, 8H); LCMS (ESI) m/z: 396.1 [M+H]+. Example 114. Preparation of N-(6-bromo-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000154_0003
Step 1: Preparation of 6-bromo-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000154_0004
To a solution of nicotinaldehyde (1.07 g, 10 mmol), 6-bromobenzo[d]thiazol-2-amine (2.29 g, 10 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.90 g, 50 mmol) in ethanol (100 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-80% ethyl acetate in hexanes to offer 6-bromo-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (0.890 g, 2.8 mmol, 28 %) as a white powder; LCMS (ESI) m/z: 320.0 [M+H]+. Step 2: Preparation of N-(6-bromo-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide
Figure imgf000155_0001
Into a solution of 6-bromo-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (200 mg, 0.60 mmol) and N,N-diisopropylethylamine (160 mg, 1.3 mmol) in dichloromethane (5 mL) was added cyclohexanecarbonyl chloride (180 mg, 1.3 mmol) at 0 °C. The reaction was warmed to room temperature with stirring over 4 hours. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane (20 mL) and washed with water (20 mL). The organic layer was concentrated under reduced pressure and purified by prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product N-(6-bromobenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)cyclohexanecarboxamide (140 mg, 0.33 mmol, 52 %) was afforded as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.57 (d, J = 1.9 Hz, 1H), 8.49 (d, J = 3.6 Hz, 1H), 8.28 (d, J = 1.9 Hz, 1H), 7.68 (d, J = 8.6 Hz, 1H), 7.63 (d, J = 8.0 Hz, 1H), 7.56 (dd, J = 8.6, 2.0 Hz, 1H), 7.37 (dd, J = 7.9, 4.8 Hz, 1H), 5.69 (s, 2H), 2.88 (t, J = 11.2 Hz, 1H), 1.65 (m, 5H), 1.42 (dd, J = 22.3, 11.0 Hz, 2H), 1.31 – 1.11 (m, 3H); LCMS (ESI) m/z: 430.0 [M+H]+. Example 115. Preparation of N-(7-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000155_0002
Step 1: Preparation of 7-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000155_0003
To a solution of nicotinaldehyde (321 mg, 3.0 mmol), 7-fluorobenzo[d]thiazol-2-amine (3m40 g, 2.0 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (296 mg, 8.0 mmol) in methanol (10 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-10% methanol in dichloromethane to offer 7-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (300 mg, 1.2 mmol, 57 %) as a yellow solid. LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of N-(7-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-3-yl)methyl]propanamide
Figure imgf000156_0001
To a solution of 7-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (130 mg, 0.50 mmol) in N,N-dimethylformamide (5 mL) was added 3-phenoxypropanoic acid (166 mg, 1.0 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (285 mg, 0.75 mmol) and triethylamine (150 mg, 1.5 mmol). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. Water (10 mL) was added to the reaction vial, and the product was extracted with 30 mL ethyl acetate. The organic layer was concentrated under reduced pressure. Crude product was purified prep-HPLC to give N-(7-fluorobenzo[d]thiazol-2-yl)-3-phenoxy-N-(pyridin-3-ylmethyl)propanamide (19.0 mg, 0.047 mmol, 9.0 %) as white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.68 (s, 1H), 8.56 (d, J = 4.8 Hz, 1H), 7.82 (d, J = 7.66 Hz, 1H), 7.64 (d, J = 8.0 Hz, 1H), 7.46 – 7.51 (m, 2H), 7.22 – 7.30 (m, 3H), 6.91- 6.96 (m, 3H), 5.72 (s, 2H), 4.32 (t, J = 5.6 Hz, 2H), 3.26 (t, J = 5.6 Hz, 2H); LCMS (ESI) m/z: 408.1 [M+H]+. Example 116. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1-methyl-1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000156_0002
Step 1: Preparation of 6-fluoro-N-((1-methyl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine To a solution of 1-methyl-1H-imidazole-4-carbaldehyde (0.785 g, 7.1 mmol), 6- fluorobenzo[d]thiazol-2-amine (1.00 g, 6.0 mmol) in toluene (30 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (0.440 g, 12 mmol) in methanol (30 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-10% methanol in dichloromethane to offer 6- fluoro-N-((1-methyl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (0.600 g, 2.3 mmol, 40 %) as a yellow solid. LCMS (ESI) m/z: 263.1 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1-methyl-1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000157_0001
To a solution of 6-fluoro-N-((1-methyl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (100 mg, 0.38 mmol) in N,N-dimethylformamide (10 mL) was added cyclohexanecarboxylic acid (73.0 mg, 0.57 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (216 mg, 1.6 mmol) and N,N-diisopropylethylamine (73.0 mg, 0.57 mmol). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. Water (10 mL) was added to the reaction vial, and the product was extracted with 30 mL ethyl acetate. The organic layer was concentrated under reduced pressure. Crude product was purified prep-HPLC to give N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1-methyl-1H-imidazol-4- yl)methyl]cyclohexanecarboxamide (36.0 mg, 0.097 mmol, 25 %) as white solid.1H NMR (300 MHz, Chloroform-d) δ 7.79 (dd, J = 8.9, 4.6 Hz, 1H), 7.60 (d, J = 1.0 Hz, 1H), 7.50 (dd, J = 8.1, 2.6 Hz, 1H), 7.18 (td, J = 8.9, 2.6 Hz, 1H), 7.09 (s, 1H), 5.44 (s, 2H), 3.66 (s, 3H), 3.10 (t, J = 10.7 Hz, 1H), 1.85 (d, J = 10.1 Hz, 4H), 1.79 – 1.53 (m, 2H), 1.39 (dd, J = 22.7, 10.4 Hz, 2H); LCMS (ESI) m/z: 373.2 [M+H]+. Example 117. Preparation of tert-butyl 6-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3- yl)methyl]carbamoyl]-2-azaspiro[3.3]heptane-2-carboxylate
Figure imgf000157_0002
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine To a solution of nicotinaldehyde (
Figure imgf000157_0003
thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of tert-butyl 6-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3-yl)methyl]carbamoyl]-2- azaspiro[3.3]heptane-2-carboxylate
Figure imgf000158_0001
To a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (0.600 g, 2.2 mmol) in dichloromethane (20 mL) were added 2-(tert-butoxycarbonyl)-2-azaspiro[3.3]heptane-6-carboxylic acid (0.805 g, 3.3 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (1.30 g, 3.3 mmol) and triethylamine (0.676 g, 6.7 mmol) at 0 °C. The reaction was warmed to room temperature over 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product tert-butyl 6-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3-yl)methyl]carbamoyl]-2- azaspiro[3.3]heptane-2-carboxylate (0.349 g, 0.71 mmol, 32 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.52 (d, J = 1.9 Hz, 1H), 8.47 (dd, J = 4.7, 1.3 Hz, 1H), 7.82 (d, J = 1.1 Hz, 1H), 7.59 (dd, J = 19.3, 8.1 Hz, 2H), 7.33 (dd, J = 7.9, 4.8 Hz, 1H), 7.26 (dd, J = 8.3, 1.6 Hz, 1H), 5.48 (s, 2H), 3.85 (s, 2H), 3.74 (s, 2H), 3.55 (p, J = 8.2 Hz, 1H), 2.70 (q, J = 7.6 Hz, 2H), 2.37 (dt, J = 21.1, 12.5 Hz, 4H), 1.35 (s, 9H), 1.21 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 493.2 [M+H]+. Example 118. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]-2- azaspiro[3.3]heptane-6-carboxamide
Figure imgf000158_0002
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]-2-azaspiro[3.3]heptane-6- carboxamide
Figure imgf000158_0003
To a solution of tert-butyl 6-((6-ethylbenzo[d]thiazol-2-yl)(pyridin-3-ylmethyl)carbamoyl)-2- azaspiro[3.3]heptane-2-carboxylate (330 mg, 0.67 mmol) in dichloromethane (12 mL) was added trifluoroacetic acid (3 mL) at 0 °C. The reaction was warmed to room temperature over 2 hours. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product N-(6- ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)-2-azaspiro[3.3]heptane-6-carboxamide (190 mg, 0.48 mmol, 73 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.58 – 8.43 (m, 2H), 7.82 (d, J = 1.1 Hz, 1H), 7.59 (dd, J = 18.3, 8.1 Hz, 2H), 7.34 (dd, J = 7.8, 4.8 Hz, 1H), 7.26 (dd, J = 8.3, 1.6 Hz, 1H), 5.49 (s, 2H), 3.96 (s, 2H), 3.85 (s, 2H), 3.58 (p, J = 8.3 Hz, 1H), 2.70 (q, J = 7.5 Hz, 2H), 2.49 – 2.28 (m, 4H), 1.22 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 393.1 [M+H]+. Example 119. Preparation of N-(1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000159_0001
Step 1: Preparation of N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000159_0002
In a reaction vial, a solution of 1H-imidazole-5-carbaldehyde (140 mg, 1.5 mmol) and 1,3- benzothiazol-2-amine (200 mg, 1.3 mmol) in dry toluene (13 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (115 mg, 3.1 mmol) in dry ethyl alcohol (13 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis, and crude product as sufficient purity to continue onto the next step. Product N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine (306 mg, 1.3 mmol, 100 %) was assumed 100% yield and afforded as a cream solid.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 7.61 (d, J = 7.4 Hz, 1H), 7.35 (d, J = 7.8 Hz, 1H), 7.17 (t, J = 6.0 Hz, 2H), 6.96 (t, J = 7.6 Hz, 1H), 6.70 (s, 1H), 4.37 (d, J = 3.1 Hz, 2H); LCMS (ESI) m/z: 231.1 [M+H]+. Step 2: Preparation of N-(1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]cyclohexanecarboxamide
Figure imgf000159_0003
In a reaction vial, N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine (306 mg, 1.3 mmol) and triethylamine (0.46 mL, 3.3 mmol) were dissolved in tetrahydrofuran (1.32 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (580 mg, 4.0 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of dichloromethane were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-50% ethyl acetate in hexanes. Product N-(1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5- yl)methyl]cyclohexanecarboxamide (127 mg, 0.38 mmol, 28 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.00 (s, 1H), 7.93 – 7.76 (m, 2H), 7.46 (dd, J = 8.1, 7.0 Hz, 1H), 7.33 (t, J = 7.6 Hz, 1H), 7.18 (s, 1H), 5.47 (s, 2H), 3.03 (t, J = 11.2 Hz, 1H), 1.84 (d, J = 11.8 Hz, 4H), 1.75 (d, J = 11.4 Hz, 1H), 1.62 (q, J = 12.2, 11.7 Hz, 2H), 1.48 – 1.22 (m, 3H); LCMS (ESI) m/z: 341.2 [M+H]+. Example 120. Preparation of N-[(pyridin-3-yl)methyl]-N-(quinazolin-2-yl)cyclohexanecarboxamide
Figure imgf000160_0001
Step 1: Preparation of N-[(pyridin-3-yl)methyl]quinazolin-2-amine
Figure imgf000160_0002
In a reaction vial, a solution of pyridine-3-carbaldehyde (0.14 mL, 1.5 mmol) and quinazolin-2- amine (200 mg, 1.4 mmol) in dry toluene (14 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen overnight. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (119 mg, 3.2 mmol) in dry ethyl alcohol (14 mL). The reaction mixture was stirred at 70 °C for 30 min and then cooled to room temperature. The product was confirmed vial UPLC analysis. The reaction was filtered over a bed of Celite and washed with ethyl acetate (2x 20 mL). Crude product was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane through 24 g of silica gel. Product N-[(pyridin-3-yl)methyl]quinazolin-2-amine (76.9 mg, 0.33 mmol, 24 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 9.00 (d, J = 0.8 Hz, 1H), 8.81 – 8.68 (m, 1H), 8.54 (dd, J = 4.8, 1.6 Hz, 1H), 7.88 – 7.51 (m, 4H), 7.39 – 7.17 (m, 2H), 5.72 (s, 1H), 4.81 (d, J = 6.1 Hz, 2H); LCMS (ESI) m/z: 237.1 [M+H]+. Step 2: Preparation of N-[(pyridin-3-yl)methyl]-N-(quinazolin-2-yl)cyclohexanecarboxamide
Figure imgf000160_0003
In a reaction vial, N-[(pyridin-3-yl)methyl]quinazolin-2-amine (76.9 mg, 0.33 mmol) was dissolved in tetrahydrofuran (1.6 mL) and cooled to 0 °C. To the solution was added triethylamine (0.068 mL, 0.49 mmol) then cyclohexanecarbonyl chloride (0.065 mL, 0.49 mmol), slowly. The reaction was stirred from 0 °C to room temperature for 1.5 hours. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane and washed with water (3x). The organic layers were pooled, dried over anhydrous sodium sulfate, filtered, and concentrated. Crude product was purified via flash column chromatography, eluting with 0-100% ethyl acetate in dichloromethane. Product N-[(pyridin-3-yl)methyl]- N-(quinazolin-2-yl)cyclohexanecarboxamide (28.4 mg, 0.082 mmol, 25 %) was afforded as a yellow solid. 1H NMR (300 MHz, Chloroform-d) δ 9.32 (s, 1H), 8.73 – 8.58 (m, 1H), 8.43 (dd, J = 5.0, 1.6 Hz, 1H), 8.07 – 7.74 (m, 4H), 7.60 (ddd, J = 8.0, 5.8, 2.4 Hz, 1H), 7.39 – 7.13 (m, 1H), 5.40 (s, 2H), 3.23 (tt, J = 11.5, 3.3 Hz, 1H), 2.14 – 1.91 (m, 2H), 1.86 – 1.44 (m, 2H), 1.43 – 1.02 (m, 2H); LCMS (ESI) m/z: 347.2 [M+H]+. Example 121. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000161_0001
Step 1: Preparation of 6-ethyl-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000161_0002
In a reaction vial, a solution of pyrimidine-5-carbaldehyde (121 mg, 1.1 mmol) and 6-ethyl-1,3- benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-ethyl-N- [(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine (45.6 mg, 0.17 mmol, 15 %) was afforded as a yellow solid.1H NMR (300 MHz, Methanol-d4) δ 9.07 (s, 1H), 8.81 (d, J = 18.9 Hz, 2H), 7.44 (s, 1H), 7.35 (d, J = 8.2 Hz, 1H), 7.21 – 7.04 (m, 1H), 4.69 (s, 2H), 2.67 (d, J = 8.4 Hz, 3H), 1.32 – 1.16 (m, 4H); LCMS (ESI) m/z: 271.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000161_0003
In a reaction vial, 6-ethyl-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine (45.6 mg, 0.17 mmol) and triethylamine (0.035 mL, 0.25 mmol) were dissolved in tetrahydrofuran (1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (36.9 mg, 0.25 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]cyclohexanecarboxamide (20.9 mg, 0.055 mmol, 33 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 9.16 (s, 1H), 8.75 (s, 2H), 7.69 (d, J = 8.4 Hz, 1H), 7.63 (dd, J = 1.8, 0.8 Hz, 1H), 7.27 (d, J = 9.1 Hz, 1H), 5.53 (s, 2H), 2.77 (q, J = 7.6 Hz, 2H), 2.74 – 2.61 (m, 1H), 1.82 (d, J = 12.9 Hz, 4H), 1.76 – 1.58 (m, 4H), 1.29 (t, J = 7.6 Hz, 5H); LCMS (ESI) m/z: 381.2 [M+H]+. Example 122. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000162_0001
Step 1: Preparation of 6-ethyl-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000162_0002
In a reaction vial, a solution of 1,3-thiazole-5-carbaldehyde (140 mg, 1.2 mmol) and 6-ethyl-1,3- benzothiazol-2-amine (219 mg, 1.2 mmol) in dry toluene (12 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (106 mg, 2.8 mmol) in dry ethyl alcohol (12 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-ethyl-N-[(1,3- thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine (240 mg, 0.87 mmol, 71 %) was afforded as a white solid. 1H NMR (300 MHz, Methanol-d4) δ 8.91 (d, J = 1.6 Hz, 1H), 7.87 (s, 1H), 7.44 (s, 1H), 7.39 (d, J = 8.3 Hz, 1H), 7.19 – 7.07 (m, 1H), 4.87 (s, 2H), 2.67 (q, J = 7.6 Hz, 2H), 1.36 – 1.14 (m, 3H); LCMS (ESI) m/z: 276.0 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000162_0003
In a reaction vial, 6-ethyl-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine (241 mg, 0.87 mmol) and triethylamine (0.18 mL, 1.3 mmol) were dissolved in tetrahydrofuran (1.7 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (190 mg, 1.3 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-50% ethyl acetate in hexanes. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5- yl)methyl]cyclohexanecarboxamide (257 mg, 0.67 mmol, 77 %) was afforded as a white solid. 1H NMR (300 MHz, Chloroform-d) δ 8.72 (s, 1H), 7.93 (s, 1H), 7.78 (d, J = 8.3 Hz, 1H), 7.63 (d, J = 1.7 Hz, 1H), 7.30 (dd, J = 8.4, 1.7 Hz, 1H), 5.70 (s, 2H), 2.88 (tt, J = 11.7, 3.3 Hz, 1H), 2.78 (q, J = 7.6 Hz, 2H), 1.93 – 1.56 (m, 8H), 1.40 – 1.20 (m, 5H); LCMS (ESI) m/z: 386.2 [M+H]+. Example 123. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000163_0001
Step 1: Preparation of 6-fluoro-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000163_0002
In a reaction vial, a solution of 1,3-thiazole-5-carbaldehyde (242 mg, 2.1 mmol) and 6-fluoro-1,3- benzothiazol-2-amine (360 mg, 2.1 mmol) in dry toluene (21 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (186 mg, 4.9 mmol) in dry ethyl alcohol (21 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-fluoro-N-[(1,3- thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine (393 mg, 1.5 mmol, 69 %) was afforded as a white solid.1H NMR (300 MHz, Methanol-d4) δ 8.91 (s, 1H), 7.93 – 7.83 (m, 1H), 7.48 – 7.36 (m, 2H), 7.12 – 6.93 (m, 1H), 4.88 – 4.79 (m, 2H); LCMS (ESI) m/z: 266.0 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000163_0003
In a reaction vial, 6-fluoro-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine (393 mg, 1.5 mmol) and triethylamine (0.31 mL, 2.2 mmol) were dissolved in tetrahydrofuran (3 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (324 mg, 2.2 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of dichloromethane were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-50% ethyl acetate in hexanes. Product N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5- yl)methyl]cyclohexanecarboxamide (256 mg, 0.68 mmol, 46 %) was afforded as a white solid. 1H NMR (300 MHz, Chloroform-d) δ 8.73 (s, 1H), 7.94 (s, 1H), 7.80 (dd, J = 8.9, 4.7 Hz, 1H), 7.50 (dd, J = 8.1, 2.6 Hz, 1H), 7.18 (td, J = 8.9, 2.6 Hz, 1H), 5.70 (s, 2H), 2.89 (tt, J = 11.2, 2.9 Hz, 1H), 1.94 – 1.72 (m, 5H), 1.74 – 1.55 (m, 2H), 1.43 – 1.23 (m, 3H); LCMS (ESI) m/z: 376.2 [M+H]+. Example 124. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperidine-4- carboxamide
Figure imgf000164_0001
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperidine-4-carboxamide
Figure imgf000164_0002
To a solution of tert-butyl 4-((6-ethylbenzo[d]thiazol-2-yl)(pyridin-3-ylmethyl)carbamoyl)piperidine- 1-carboxylate (150 mg, 0.31 mmol) in dichloromethane (12 mL) at 0 °C was added trifluoroacetic acid (3 mL). The reaction was warmed to room temperature over 2 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C18 21*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product N-(6- ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)piperidine-4-carboxamide (36.3 mg, 0.090 mmol, 31 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.59 (s, 1H), 8.49 (d, J = 4.1 Hz, 1H), 7.82 (s, 1H), 7.65 (d, J = 8.3 Hz, 2H), 7.36 (dd, J = 7.8, 4.8 Hz, 1H), 7.28 (dd, J = 8.4, 1.4 Hz, 1H), 5.68 (s, 2H), 3.21 (d, J = 12.5 Hz, 3H), 2.82 (s, 2H), 2.71 (q, J = 7.5 Hz, 2H), 1.79 (s, 4H), 1.22 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 381.1 [M+H]+. Example 125. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]oxolane-2- carboxamide
Figure imgf000164_0003
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000164_0004
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]oxolane-2-carboxamide
Figure imgf000165_0001
To a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (120 mg, 0.45 mmol) in dichloromethane (15 mL) were added tetrahydrofuran-2-carboxylic acid (77.7 mg, 0.67 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (255 mg, 0.67 mmol) and triethylamine (136 mg, 1.4 mmol) at 0 °C. The reaction was warmed to room temperature over 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep- HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C18 21*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product N-(6- ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)tetrahydrofuran-2-carboxamide (68.0 mg, 0.18 mmol, 41 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.55 (d, J = 1.7 Hz, 1H), 8.46 (d, J = 3.5 Hz, 1H), 7.83 (d, J = 1.2 Hz, 1H), 7.63 (dd, J = 8.0, 5.1 Hz, 2H), 7.34 (dd, J = 7.9, 4.8 Hz, 1H), 7.27 (dd, J = 8.3, 1.7 Hz, 1H), 5.64 (dd, J = 41.7, 17.2 Hz, 2H), 4.94 (t, J = 6.5 Hz, 1H), 4.01 – 3.55 (m, 2H), 2.71 (q, J = 7.6 Hz, 2H), 2.11 (dd, J = 14.3, 6.9 Hz, 2H), 2.01 – 1.68 (m, 2H), 1.22 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 368.0 [M+H]+. Example 126. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]pyrrolidine-1- carboxamide
Figure imgf000165_0002
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000165_0003
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]pyrrolidine-1-carboxamide
Figure imgf000166_0001
To a solution of triphosgene (65.3 mg, 0.22 mmol) in dichloromethane (10 mL) was added a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (120 mg, 0.45 mmol) and pyridine (142 mg, 1.8 mmol) in dichloromethane (5 mL) at -60 °C under argon. The reaction was warmed to 0 °C over 2 hours. Then a solution of pyrrolidine (47.6 mg, 0.67 mmol) and pyridine (142 mg, 1.8 mmol) in dichloromethane (6 mL) was added at -60 °C. The resulting mixture was warmed to room temperature with stirring over 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product N-(6-ethylbenzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)pyrrolidine-1-carboxamide (26.2 mg, 0.070 mmol, 16 %) was afforded as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.57 (t, J = 8.6 Hz, 1H), 8.43 (dt, J = 12.2, 6.1 Hz, 1H), 7.80 – 7.65 (m, 2H), 7.55 (d, J = 8.2 Hz, 1H), 7.42 – 7.29 (m, 1H), 7.20 (dd, J = 8.3, 1.6 Hz, 1H), 5.18 (s, 2H), 3.37 (s, 4H), 2.75 – 2.59 (m, 2H), 1.80 (t, J = 6.5 Hz, 4H), 1.21 (dt, J = 15.1, 7.5 Hz, 3H); LCMS (ESI) m/z: 367.1 [M+H]+. Example 127. Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000166_0002
Step 1: Preparation of 6-chloro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000166_0003
To a solution of nicotinaldehyde (0.890 g, 8.4 mmol), 6-ethylbenzo[d]thiazol-2-amine (1.00 g, 5.6 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (0.638 g, 17 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-chloro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (0.700 g, 2.5 mmol, 45 %) as a white solid. LCMS (ESI) m/z: 276.1 [M+H]+. Step 2: Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide
Figure imgf000167_0001
To a solution of 6-chloro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (110 mg, 0.39 mmol) and triethylamine (78.0 mg, 0.78 mmol) in dichloromethane (5 mL) was added cyclohexanecarbonyl chloride (85.0 mg, 0.59 mmol). The reaction was stirred at room temperature overnight. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate). Product N-(6-chlorobenzo[d]thiazol-2-yl)-N-(pyridin- 3-ylmethyl)cyclohexanecarboxamide (31.3 mg, 0.080 mmol, 21 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.69 (d, J = 1.8 Hz, 1H), 8.65 – 8.54 (m, 1H), 8.16 (d, J = 2.1 Hz, 1H), 7.87 (d, J = 8.0 Hz, 1H), 7.72 (d, J = 8.6 Hz, 1H), 7.55 (dd, J = 7.8, 5.1 Hz, 1H), 7.44 (dd, J = 8.6, 2.2 Hz, 1H), 5.71 (s, 2H), 2.88 (t, J = 11.1 Hz, 1H), 1.67 (t, J = 11.7 Hz, 4H), 1.43 (d, J = 11.0 Hz, 2H), 1.24 (d, J = 14.3 Hz, 3H); LCMS (ESI) m/z: 386.1 [M+H]+. Example 128. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(phenylamino)-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000167_0002
Step 1: Preparation of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000167_0003
To a solution of nicotinaldehyde (481 mg, 4.5 mmol), 5-fluorobenzo[d]thiazol-2-amine (500 mg, 3.0 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (342 mg, 9.0 mmol) in ethanol (10 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (600 mg, 2.3 mmol, 77 %) as a white solid. LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(phenylamino)-N-[(pyridin-3- yl)methyl]propanamide
Figure imgf000168_0001
To a solution of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (300 mg, 1.2 mmol) in N,N-dimethylformamide (15 mL) was added 3-(phenylamino)propanoic acid (286 mg, 1.7 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (661 mg, 1.7 mmol) and N,N-diisopropylethylamine (223 mg, 1.7 mmol). The reaction mixture was stirred at 80 °C for 16 hours. The product was indicated present via UPLC analysis. Water (20 mL) was added to the reaction, which was then extracted with 2x 20 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified prep-HPLC to give N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(phenylamino)-N-[(pyridin-3- yl)methyl]propenamide (11.0 mg, 0.027 mmol, 3.0 %) as white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.56 (s, 1H), 8.49 (dd, J = 23.7, 20.0 Hz, 1H), 8.05 (dd, J = 8.7, 5.5 Hz, 1H), 7.60 (dd, J = 16.1, 5.9 Hz, 2H), 7.32 (dd, J = 7.8, 4.8 Hz, 1H), 7.28 – 7.13 (m, 1H), 7.05 (t, J = 7.8 Hz, 2H), 6.60 – 6.42 (m, 3H), 5.62 (d, J = 11.7 Hz, 3H), 3.63 – 3.33 (m, 2H), 2.97 (t, J = 6.6 Hz, 2H); LCMS (ESI) m/z: 407.0 [M+H]+. Example 129. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000168_0002
Step 1: Preparation of 6-ethyl-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000168_0003
In a reaction vial, a solution of 1H-imidazole-4-carbaldehyde (107 mg, 1.1 mmol) and 6-ethyl-1,3- benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The crude product was not purified. Product 6-ethyl-N-[(1H-imidazol-5- yl)methyl]-1,3-benzothiazol-2-amine (367 mg, 1.4 mmol, 126 %) was afforded as a brown solid.1H NMR (300 MHz, Methanol-d4) δ 7.65 (s, 1H), 7.45 – 7.29 (m, 2H), 7.15 – 7.01 (m, 2H), 4.56 (s, 2H), 2.67 (q, J = 7.5 Hz, 2H), 1.24 (t, J = 7.5 Hz, 3H); LCMS (ESI) m/z: 259.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000169_0001
In a reaction vial, 6-ethyl-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine (289 mg, 1.1 mmol) and triethylamine (0.23 mL, 1.7 mmol) were dissolved in tetrahydrofuran (2.2 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (243 mg, 1.7 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of dichloromethane were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5- yl)methyl]cyclohexanecarboxamide (255 mg, 0.69 mmol, 62 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 7.78 (d, J = 8.3 Hz, 1H), 7.63 (d, J = 1.7 Hz, 1H), 7.58 (d, J = 1.0 Hz, 1H), 7.30 (dd, J = 8.3, 1.8 Hz, 1H), 7.08 (s, 1H), 5.42 (s, 2H), 3.05 (t, J = 11.5 Hz, 1H), 2.78 (q, J = 7.6 Hz, 2H), 1.85 (d, J = 11.0 Hz, 4H), 1.76 (d, J = 9.1 Hz, 1H), 1.62 (q, J = 12.8, 12.2 Hz, 2H), 1.30 (t, J = 7.6 Hz, 3H), 1.47 – 1.15 (m, 3H); LCMS (ESI) m/z: 369.2 [M+H]+. Example 130. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridazin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000169_0002
Step 1: Preparation of 6-ethyl-N-[(pyridazin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000169_0003
In a reaction vial, a solution of pyridazine-3-carbaldehyde (121 mg, 1.1 mmol) and 6-ethyl-1,3- benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-ethyl-N- [(pyridazin-3-yl)methyl]-1,3-benzothiazol-2-amine (91.1 mg, 0.34 mmol, 30 %) was afforded as a brown solid.1H NMR (300 MHz, Methanol-d4) δ 9.08 (d, J = 4.9 Hz, 1H), 7.78 (d, J = 8.5 Hz, 1H), 7.68 (dd, J = 8.6, 5.0 Hz, 1H), 7.43 (s, 1H), 7.31 (d, J = 8.3 Hz, 1H), 7.09 (d, J = 8.2 Hz, 1H), 4.94 (s, 2H), 2.65 (q, J = 7.6 Hz, 2H), 1.28 – 1.16 (m, 3H); LCMS (ESI) m/z: 271.1 [M+H]+ Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridazin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000170_0001
In a reaction vial, 6-ethyl-N-[(pyridazin-3-yl)methyl]-1,3-benzothiazol-2-amine (91.1 mg, 0.34 mmol) and triethylamine (0.070 mL, 0.50 mmol) were dissolved in tetrahydrofuran (1.1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (73.8 mg, 0.50 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of dichloromethane were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridazin-3- yl)methyl]cyclohexanecarboxamide (36.8 mg, 0.097 mmol, 29 %) was afforded as a white solid. 1H NMR (300 MHz, Chloroform-d) δ 9.13 (dd, J = 4.9, 1.6 Hz, 1H), 7.68 (d, J = 8.4 Hz, 1H), 7.63 (d, J = 1.7 Hz, 1H), 7.56 (dd, J = 8.5, 1.6 Hz, 1H), 7.41 (dd, J = 8.5, 4.9 Hz, 1H), 7.29 – 7.23 (m, 1H), 5.91 (s, 2H), 3.02 (tt, J = 11.1, 2.8 Hz, 1H), 2.76 (q, J = 7.6 Hz, 2H), 1.88 – 1.47 (m, 5H), 1.35 – 1.23 (m, 8H); LCMS (ESI) m/z: 381.2 [M+H]+. Example 131. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(2-fluorophenoxy)-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000170_0002
Step 1: Preparation of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000170_0003
To a solution of nicotinaldehyde (481 mg, 4.5 mmol), 5-fluorobenzo[d]thiazol-2-amine (500 mg, 3.0 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (342 mg, 9.0 mmol) in methanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (600 mg, 2.3 mmol, 77 %) as a white solid. LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(2-fluorophenoxy)-N-[(pyridin-3- yl)methyl]propanamide
Figure imgf000171_0001
To a solution of 3-(2-fluorophenoxy)propanoic acid (107 mg, 0.58 mmol) in dichloromethane (15 mL) were added 1-hydroxybenzotriazole (86.4 mg, 0.64 mmol) and 1,3-dicyclohexylcarbodiimide (132 mg, 0.64 mmol). The mixture was stirred at 0 °C for 2 hours. Then a solution of 5-fluoro-N-(pyridin-3- ylmethyl)benzo[d]thiazol-2-amine (150 mg, 0.58 mmol) in pyridine (45.8 mg, 0.58 mmol) and N,N- dimethylformamide (5 mL) was added. The reaction mixture was stirred at 40 °C for 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product N-(5-fluorobenzo[d]thiazol-2-yl)-3-(2-fluorophenoxy)-N-(pyridin-3- ylmethyl)propanamide (28.9 mg, 0.070 mmol, 12 %) was afforded as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.61 (s, 1H), 8.50 (d, J = 3.9 Hz, 1H), 8.05 (dd, J = 8.6, 5.5 Hz, 1H), 7.77 – 7.55 (m, 2H), 7.36 (dd, J = 7.7, 4.9 Hz, 1H), 7.30 – 7.01 (m, 4H), 7.05 – 6.86 (m, 1H), 5.72 (d, J = 24.1 Hz, 2H), 4.40 (t, J = 5.7 Hz, 2H), 3.27 (t, J = 5.7 Hz, 2H); LCMS (ESI) m/z: 426.0 [M+H]+. Example 132. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(3-fluorophenoxy)-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000171_0002
Step 1: Preparation of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000171_0003
To a solution of nicotinaldehyde (481 mg, 4.5 mmol), 5-fluorobenzo[d]thiazol-2-amine (500 mg, 3.0 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (342 mg, 9.0 mmol) in methanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (600 mg, 2.3 mmol, 77 %) as a white solid. LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(3-fluorophenoxy)-N-[(pyridin-3- yl)methyl]propanamide
Figure imgf000172_0001
To a solution of 3-(3-fluorophenoxy)propanoic acid (107 mg, 0.58 mmol) in dichloromethane (15 mL) were added 1-hydroxybenzotriazole (86.4 mg, 0.64 mmol) and 1,3-dicyclohexylcarbodiimide (132 mg, 0.64 mmol). The mixture was stirred at 0 °C for 2 hours. Then a solution of 5-fluoro-N-(pyridin-3- ylmethyl)benzo[d]thiazol-2-amine (150 mg, 0.58 mmol) in pyridine (45.8 mg, 0.58 mmol) and N,N- dimethylformamide (5 mL) was added. The reaction mixture was stirred at 40 °C for 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product N-(5-fluorobenzo[d]thiazol-2-yl)-3-(3-fluorophenoxy)-N-(pyridin-3- ylmethyl)propanamide (26.5 mg, 0.060 mmol, 11 %) was afforded as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.59 (d, J = 1.8 Hz, 1H), 8.49 (dd, J = 6.6, 5.3 Hz, 1H), 8.05 (dd, J = 8.8, 5.5 Hz, 1H), 7.68 (d, J = 7.9 Hz, 1H), 7.61 (dd, J = 10.1, 2.4 Hz, 1H), 7.38 (dd, J = 7.8, 4.7 Hz, 1H), 7.34 – 7.18 (m, 2H), 6.78 (ddd, J = 13.5, 6.6, 2.0 Hz, 3H), 5.72 (d, J = 24.4 Hz, 2H), 4.33 (t, J = 5.8 Hz, 2H), 3.22 (t, J = 5.8 Hz, 2H); LCMS (ESI) m/z: 426.1 [M+H]+. Example 133. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(4-fluorophenoxy)-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000172_0002
Step 1: Preparation of 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000172_0003
To a solution of nicotinaldehyde (481 mg, 4.5 mmol), 5-fluorobenzo[d]thiazol-2-amine (500 mg, 3.0 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (342 mg, 9.0 mmol) in methanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 5-fluoro-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (600 mg, 2.3 mmol, 77 %) as a white solid. LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(4-fluorophenoxy)-N-[(pyridin-3- yl)methyl]propanamide
Figure imgf000173_0001
To a solution of 3-(4-fluorophenoxy)propanoic acid (107 mg, 0.58 mmol) in dichloromethane (15 mL) were added 1-hydroxybenzotriazole (86.4 mg, 0.64 mmol) and 1,3-dicyclohexylcarbodiimide (132 mg, 0.64 mmol). The mixture was stirred at 0 °C for 2 hours. Then a solution of 5-fluoro-N-(pyridin-3- ylmethyl)benzo[d]thiazol-2-amine (150 mg, 0.58 mmol) in pyridine (45.8 mg, 0.58 mmol) and N,N- dimethylformamide (5 mL) was added. The reaction mixture was stirred at 40 °C for 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product N-(5-fluorobenzo[d]thiazol-2-yl)-3-(4-fluorophenoxy)-N-(pyridin-3- ylmethyl)propanamide (29.3 mg, 0.070 mmol, 12 %) was afforded as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.60 (d, J = 1.6 Hz, 1H), 8.50 (d, J = 3.5 Hz, 1H), 8.05 (dd, J = 8.7, 5.5 Hz, 1H), 7.68 (d, J = 8.0 Hz, 1H), 7.61 (dd, J = 10.0, 2.4 Hz, 1H), 7.38 (dd, J = 7.8, 4.8 Hz, 1H), 7.24 (td, J = 9.1, 2.4 Hz, 1H), 7.10 (t, J = 8.8 Hz, 2H), 7.01 – 6.84 (m, 2H), 5.69 (s, 2H), 4.29 (t, J = 5.8 Hz, 2H), 3.21 (t, J = 5.8 Hz, 2H); LCMS (ESI) m/z: 426.1 [M+H]+. Example 134. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]benzamide
Figure imgf000173_0002
Step 1: Preparation of 6-ethyl-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000173_0003
To a solution of 1H-imidazole-4-carbaldehyde (0.799 g, 8.3 mmol), 6-ethylbenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (0.308 g, 8.3 mmol) in methanol (50 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-[(1H-imidazol-5- yl)methyl]-1,3-benzothiazol-2-amine (1.20 g, 4.6 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 259.0 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]benzamide
Figure imgf000174_0001
To a solution of N-((1H-imidazol-4-yl)methyl)-6-ethylbenzo[d]thiazol-2-amine (200 mg, 0.76 mmol) in N,N-dimethylformamide (5 mL) was added benzoic acid (186 mg, 1.5 mmol), N-[(dimethylamino)-1H- 1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (577 mg, 1.5 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. Water (20 mL) was added, then the reaction was extracted with 2x 20 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified prep-HPLC to give N- (6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]benzamide (95.0 mg, 0.26 mmol, 43%) as white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.57 – 8.45 (m, 2H), 8.04 (dd, J = 8.8, 5.5 Hz, 1H), 7.59 (dd, J = 10.1, 2.6 Hz, 2H), 7.43 – 7.25 (m, 1H), 7.25 – 7.07 (m, 6H), 5.59 (s, 2H), 2.83 – 2.52 (m, 4H), 2.13 – 1.63 (m, 2H); LCMS (ESI) m/z: 363.0 [M+H]+. Example 135. Preparation of 4-cyano-N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol- 2-yl)benzamide
Figure imgf000174_0002
Step 1: Preparation of 6-methoxy-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000174_0003
To a solution of 1H-imidazole-4-carbaldehyde (0.799 g, 8.3 mmol), 6-methoxybenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (0.308 g, 8.3 mmol) in methanol (50 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-methoxy-N-[(1H-imidazol- 5-yl)methyl]-1,3-benzothiazol-2-amine (1.00 g, 3.8 mmol, 68 %) as a white solid. LCMS (ESI) m/z: 261.0 [M+H]+. Step 2: Preparation of 4-cyano-N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2- yl)benzamide To a solution of N-((1H-imidazol-4-yl)methyl)-6-methoxybenzo[d]thiazol-2-amine (200 mg, 0.76 mmol) in N,N-dimethylformamide (5 mL) was added 4-cyanobenzoic acid (223 mg, 1.5 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (577 mg, 1.5 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. Water (20 mL) was added, then the reaction was extracted using 2x 20 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified prep-HPLC to give 4-cyano-N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3- benzothiazol-2-yl)benzamide (80.0 mg, 0.21 mmol, 34 %) as white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.57 – 8.45 (m, 2H), 8.04 (dd, J = 8.8, 5.5 Hz, 1H), 7.59 (dd, J = 10.1, 2.6 Hz, 2H), 7.43 – 7.25 (m, 1H), 7.25 – 7.07 (m, 6H), 5.59 (s, 2H), 2.83 – 2.52 (m, 4H), 2.13 – 1.63 (m, 2H); LCMS (ESI) m/z: 390.0 [M+H]+. Example 136. Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-4-cyano-N-[(1H-imidazol-4- yl)methyl]benzamide
Figure imgf000175_0001
Step 1: Preparation of 6-chloro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine To a solution of 1H-imidazole-4-carbaldehyde (0.782 g, 8.2 mmol), 6-chlorobenzo[d]thiazol-2- amine (1.00 g, 5.4 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (0.301 g, 8.2 mmol) in methanol (50 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-10% methanol in dichloromethane to offer 6-chloro-N-[(1H-imidazol-5- yl)methyl]-1,3-benzothiazol-2-amine (0.800 g, 3.0 mmol, 60 %) as a white solid. LCMS (ESI) m/z: 265.0 [M+H]+. Step 2: Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-4-cyano-N-[(1H-imidazol-4-yl)methyl]benzamide
Figure imgf000175_0002
To a solution of 6-chloro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine (200 mg, 0.75 mmol) in N,N-dimethylformamide (5 mL) was added 4-cyanobenzoic acid (223 mg, 1.5 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (577 mg, 1.5 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. Water (20 mL) was added, then the reaction was extracted using 2x 20 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified prep-HPLC to give N-(6-chloro-1,3-benzothiazol-2-yl)-4-cyano-N-[(1H-imidazol-4- yl)methyl]benzamide (80.0 mg, 0.20 mmol, 34 %) as white solid.1H NMR (400 MHz, Dimethylsulfoxide- d6) δ 8.57 – 8.45 (m, 2H), 8.04 (dd, J = 8.8, 5.5 Hz, 1H), 7.59 (dd, J = 10.1, 2.6 Hz, 2H), 7.43 – 7.25 (m, 1H), 7.25 – 7.07 (m, 6H), 5.59 (s, 2H), 2.83 – 2.52 (m, 4H), 2.13 – 1.63 (m, 2H); LCMS (ESI) m/z: 393.0 [M+H]+. Example 137. Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3- yl)methyl]acetamide
Figure imgf000176_0001
Step 1: Preparation of 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000176_0002
To a solution of nicotinaldehyde (1.19 g, 11 mmol), 6-methoxybenzo[d]thiazol-2-amine (2.00 g, 11 mmol) in toluene (10 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (2.11 g, 56 mmol) in methanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol- 2-amine (1.10 g, 4.1 mmol, 36 %) as a white solid. LCMS (ESI) m/z: 272.1 [M+H]+. Step 2: Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3-yl)methyl]acetamide
Figure imgf000176_0003
In a reaction vial, 6-methoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (200 mg, 0.74 mmol) and 2-phenylacetic acid (120 mg, 0.89 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (420 mg, 1.1 mmol) and N,N- diisopropylethylamine (286 mg, 2.2 mmol) were dissolved in dichloromethane (5 mL). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. Water (20 mL) was added, then the reaction was extracted using 2x 20 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by flash chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane to give N- (6-methoxybenzo[d]thiazol-2-yl)-2-phenyl-N-(pyridin-3-ylmethyl)acetamide (100 mg, 0.26 mmol, 35 %) as a yellow solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.57 (s,1H ), 8.48-8.49 (d, J = 3.6 Hz, 1H), 7.63-7.65 (d, J = 9.2 Hz, 2H), 7.58-7.58 (d, J = 2.8 Hz, 1H), 7.20-7.37 (m, 6H), 7.00-7.03 (m,1H), 5.68 (s,2H), 4.09 (s, 2H), 3.80 (s, 3H); LCMS (ESI) m/z: 390.0 [M+H]+. Example 138. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-2- phenylacetamide
Figure imgf000177_0001
Step 1: Preparation of 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000177_0002
To a solution of 1H-imidazole-4-carbaldehyde (2.50 g, 27 mmol), 6-fluorobenzo[d]thiazol-2-amine (3.00 g, 18 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.30 g, 36 mmol) in methanol (50 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3- benzothiazol-2-amine (2.50 g, 10 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 249.1 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-2-phenylacetamide
Figure imgf000177_0003
In a reaction vial, 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine (200 mg, 0.81 mmol) and 2-phenylacetic acid (132 mg, 0.97 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin- 1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (459 mg, 1.2 mmol) and N,N- diisopropylethylamine (312 mg, 2.4 mmol) were dissolved in dichloromethane (5 mL). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. Water (20 mL) was added, then the reaction was extracted using 2x 20 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by flash chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane to give N- (6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-2-phenylacetamide (120 mg, 0.33 mmol, 40 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 12.08 (s, 1H), 8.31 (s, 1H), 7.82-7.89 (m, 2H), 7.65 (s, 1H), 7.16-7.38 (m, 6H), 7.16 (s,1H), 5.48 (s, 2H), 4.51 (s, 2H); LCMS (ESI) m/z: 367.0 [M+H]+. Example 139. Preparation of N-[(1H-imidazol-5-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2-yl)-2- phenylacetamide
Figure imgf000178_0001
Step 1: Preparation of 6-methoxy-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000178_0002
To a solution of 1H-imidazole-4-carbaldehyde (0.799 g, 8.3 mmol), 6-methoxybenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (0.308 g, 8.3 mmol) in methanol (50 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-methoxy-N-[(1H-imidazol- 5-yl)methyl]-1,3-benzothiazol-2-amine (1.00 g, 3.8 mmol, 68 %) as a white solid. LCMS (ESI) m/z: 261.0 [M+H]+. Step 2: Preparation of N-[(1H-imidazol-5-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2-yl)-2- phenylacetamide
Figure imgf000178_0003
In a reaction vial, N-((1H-imidazol-5-yl)methyl)-6-methoxybenzo[d]thiazol-2-amine (230 mg, 0.88 mmol) and 2-phenylacetic acid (144 mg, 1.1 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (504 mg, 1.3 mmol) and N,N- diisopropylethylamine (342 mg, 2.7 mmol) were dissolved in dichloromethane (5 mL). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. Water (20 mL) was added, then the reaction was extracted using 2x 20 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by flash chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane to give N- [(1H-imidazol-5-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2-yl)-2-phenylacetamide (30.0 mg, 0.079 mmol, 10%) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 12.07 (s, 1H), 8.38 (s, 1H), 7.71-7.73 (d, J=8.8Hz, 1H), 7.64 (s, 1H), 7.53 (s, 1H), 7.25-7.37 (m, 5H), 7.12 (s, 1H), 7.01-7.04 (m, 1H), 5.46 (s, 1H), 4.75 (s, 2H), 3.80 (s, 3H); LCMS (ESI) m/z: 379.0 [M+H]+. Example 140. Preparation of 4-chloro-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]benzamide
Figure imgf000179_0001
Step 1: Preparation of 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000179_0002
In a round bottom flask equipped with a stir bar, a solution of pyridine-3-carbaldehyde (0.63 mL, 6.7 mmol) and 6-ethyl-1,3-benzothiazol-2-amine (1.20 g, 6.7 mmol) in dry toluene (34 mL) was stirred at 105°C with activated 4 Å molecular sieves under nitrogen overnight. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (1.27 g, 34 mmol) in dry ethyl alcohol (34 mL). The reaction mixture was stirred at 70 °C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with ethyl acetate. The filtrate was concentrated then diluted with dichloromethane and filtered again. The filtrate was purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane through a 40 g column of silica gel. Product 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3- benzothiazol-2-amine (0.915 g, 3.4 mmol, 51 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.72 – 8.50 (m, 2H), 7.76 (dddd, J = 7.9, 2.4, 1.7, 0.8 Hz, 1H), 7.50 – 7.37 (m, 2H), 7.35 – 7.22 (m, 1H), 7.14 (dt, J = 8.3, 1.2 Hz, 1H), 6.24 (s, 1H), 4.68 (s, 2H), 2.70 (q, J = 7.6 Hz, 2H), 1.26 (td, J = 7.6, 0.7 Hz, 3H); LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of 4-chlorobenzoyl chloride
Figure imgf000179_0003
In a reaction vial, 4-chlorobenzoic acid (300 mg, 1.9 mmol) was dissolved in dichloromethane (9.5 mL) and cooled to 0 °C. To the reaction was added oxalyl chloride (0.81 mL, 9.5 mmol) slowly. A few drops of N,N-dimethylformamide were added as a catalyst and the reaction bubbled. The reaction was then heated to 40 °C and stirred overnight. The product was indicated present via crude NMR analysis. The reaction was cooled to room temperature and concentrated under reduced pressure. Crude product was indicated pure enough to continue to next step via NMR and assumed 100% yield. Product 4- chlorobenzoyl chloride (334 mg, 1.9 mmol, 100 %) was afforded as a yellow oil. 1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 8.02 (s, 1H), 3.14 – 2.41 (m, 4H); LCMS data unavailable. Step 3: Preparation of 4-chloro-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]benzamide
Figure imgf000180_0001
In a reaction vial, 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (400 mg, 1.5 mmol) was dissolved in tetrahydrofuran (7.4 mL) and cooled to 0 °C. To the solution was added triethylamine (0.31 mL, 2.2 mmol) then 4-chlorobenzoyl chloride (334 mg, 1.9 mmol) was added, slowly. The reaction was stirred from 0 °C to room temperature for 1.5 hours. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane and washed with water (3x). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was dissolved with ethyl acetate and filtered again. Crude product was purified via flash column chromatography eluting with 0-100% hexanes/ethyl acetate through 40 g of silica gel. Product 4-chloro-N-(6-ethyl-1,3-benzothiazol-2- yl)-N-[(pyridin-3-yl)methyl]benzamide (62.6 mg, 0.15 mmol, 10 %) was afforded as a pale yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.59 – 8.37 (m, 2H), 7.74 (d, J = 8.4 Hz, 1H), 7.69 – 7.54 (m, 2H), 7.49 – 7.17 (m, 6H), 5.51 (s, 2H), 2.78 (q, J = 7.6 Hz, 2H), 1.31 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 408.2 [M+H]+. Example 141. Preparation of N-[(pyridin-3-yl)methyl]-N-(quinoxalin-2-yl)cyclohexanecarboxamide
Figure imgf000180_0002
Step 1: Preparation of N-[(pyridin-3-yl)methyl]quinoxalin-2-amine
Figure imgf000180_0003
In a reaction vial, a solution of pyridine-3-carbaldehyde (0.097 mL, 1.0 mmol) and quinoxalin-2- amine (150 mg, 1.0 mmol) in dry toluene (5.2 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen overnight. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (194 mg, 5.2 mmol) in dry ethyl alcohol (5.2 mL). The reaction mixture was stirred at 70 °C for 30 minutes and then cooled to room temperature. The reaction was filtered over a bed of Celite and washed with ethyl acetate (2x 20 mL). The filtrate was concentrated then diluted with dichloromethane and filtered again. The filtrate was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane. Product N-[(pyridin-3-yl)methyl]quinoxalin-2-amine (106 mg, 0.45 mmol, 44 %) was afforded as a beige solid.1H NMR (300 MHz, Chloroform-d) δ 8.71 (d, J = 2.3 Hz, 1H), 8.56 (dd, J = 4.9, 1.7 Hz, 1H), 8.25 (s, 1H), 7.90 (dd, J = 8.2, 1.5 Hz, 1H), 7.83 – 7.69 (m, 2H), 7.60 (ddd, J = 8.4, 6.9, 1.5 Hz, 1H), 7.43 (ddd, J = 8.3, 7.0, 1.5 Hz, 1H), 7.38 – 7.20 (m, 1H), 5.29 (s, 1H), 4.80 (d, J = 5.8 Hz, 2H); LCMS (ESI) m/z: 237.1 [M+H]+. Step 2: Preparation of N-[(pyridin-3-yl)methyl]-N-(quinoxalin-2-yl)cyclohexanecarboxamide
Figure imgf000181_0001
In a reaction vial, N-[(pyridin-3-yl)methyl]quinoxalin-2-amine (107 mg, 0.45 mmol) was added. was dissolved in tetrahydrofuran (2.2 mL) and cooled to 0 °C. To the solution was added triethylamine (0.094 mL, 0.67 mmol) then cyclohexanecarbonyl chloride (98.5 mg, 0.67 mmol) was added, slowly. The reaction was stirred from 0 °C to room temperature for 1.5 hours. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane and washed with water (3x). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Crude product was purified via flash column chromatography eluting with 0-100% hexanes/ethyl acetate through 24 g of silica gel. Product N-[(pyridin-3-yl)methyl]-N-(quinoxalin-2-yl)cyclohexanecarboxamide (105 mg, 0.30 mmol, 68 %) was afforded as a yellow oil.1H NMR (300 MHz, Chloroform-d) δ 8.81 (s, 1H), 8.72 – 8.41 (m, 2H), 8.19 – 7.92 (m, 2H), 7.89 – 7.63 (m, 3H), 7.38 – 7.17 (m, 1H), 5.29 (d, J = 12.7 Hz, 2H), 2.51 (tt, J = 11.3, 3.4 Hz, 1H), 1.94 – 1.46 (m, 7H), 1.38 – 0.92 (m, 3H); LCMS (ESI) m/z: 347.2 [M+H]+. Example 142. Preparation of (1s,3r)-N-(6-ethyl-1,3-benzothiazol-2-yl)-3-methyl-N-[(pyridin-3- yl)methyl]cyclobutane-1-carboxamide
Figure imgf000181_0002
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000181_0003
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of (1s,3r)-N-(6-ethyl-1,3-benzothiazol-2-yl)-3-methyl-N-[(pyridin-3- yl)methyl]cyclobutane-1-carboxamide
Figure imgf000182_0001
To a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (120 mg, 0.45 mmol) in dichloromethane (15 mL) were added 3-methylcyclobutanecarboxylic acid (76.4 mg, 0.67 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (255 mg, 0.67 mmol) and triethylamine (136 mg, 1.4 mmol). The reaction was stirred at 10 °C for 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was further purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Racemic product N-(6- ethylbenzo[d]thiazol-2-yl)-3-methyl-N-(pyridin-3-ylmethyl)cyclobutanecarboxamide (80.0 mg, 0.22 mmol, 49 %) was afforded as a white solid.80 mg of this product was sent to chiral separation (the crude sample was dissolved in 35 mL methanol. AD 20*250 mm column. The mobile phase was 0.2% methanol/ammonia) to afford two isomers. Product (1s,3r)-N-(6-ethylbenzo[d]thiazol-2-yl)-3-methyl-N- (pyridin-3-ylmethyl)cyclobutanecarboxamide (14.7 mg, 0.040 mmol, 8.9 %) was afforded as white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.57 – 8.30 (m, 2H), 7.82 (s, 1H), 7.58 (dd, J = 22.5, 8.1 Hz, 2H), 7.43 – 7.32 (m, 1H), 7.28 – 7.14 (m, 1H), 5.51 (s, 2H), 3.47 (p, J = 8.8 Hz, 1H), 2.70 (q, J = 7.5 Hz, 2H), 2.35 – 2.13 (m, 3H), 1.90 – 1.75 (m, 2H), 1.33 – 1.09 (m, 3H), 0.98 (d, J = 6.3 Hz, 3H); LCMS (ESI) m/z: 366.0 [M+H]+. Example 143. Preparation of N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2- yl)cyclohexanecarboxamide
Figure imgf000182_0002
Step 1: Preparation of 6-methoxy-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000182_0003
To a solution of 1H-imidazole-4-carbaldehyde (0.799 g, 8.3 mmol), 6-methoxybenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (0.308 g, 8.3 mmol) in methanol (50 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-methoxy-N-[(1H-imidazol- 5-yl)methyl]-1,3-benzothiazol-2-amine (1.00 g, 3.8 mmol, 68 %) as a white solid. LCMS (ESI) m/z: 261.0 [M+H]+. Step 2: Preparation of N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2- yl)cyclohexanecarboxamide
Figure imgf000183_0001
In a reaction vial, N-((1H-imidazol-5-yl)methyl)-6-methoxybenzo[d]thiazol-2-amine (230 mg, 0.88 mmol) and cyclohexanecarboxylic acid (135 mg, 1.1 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5- b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (504 mg, 1.3 mmol) and N,N-diisopropylethylamine (342 mg, 2.6 mmol) were dissolved in dichloromethane (5 mL). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. Water (20 mL) was added, then the reaction was extracted using 2x 20 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by flash chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane to give N- [(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2-yl)cyclohexanecarboxamide (48.0 mg, 0.13 mmol, 15 %) as a yellow solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 11.96 (s, 1H), 7.68-7.70 (d, J = 8.8 Hz, 1H), 7.56 (s,1H), 7.52 (s, 1H), 7.00-7.04 (m, 2H), 5.42 (s, 2H), 3.80 (s, 3H), 1.66-1.83 (m, 6H), 1.35-1.48 (m, 2H), 1.20-1.28 (m, 3H); LCMS (ESI) m/z: 371.0 [M+H]+. Example 144. Preparation of N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2- yl)benzamide
Figure imgf000183_0002
Step 1: Preparation of 6-methoxy-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000183_0003
To a solution of 1H-imidazole-4-carbaldehyde (0.799 g, 8.3 mmol), 6-methoxybenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (0.308 g, 8.3 mmol) in methanol (50 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-methoxy-N-[(1H-imidazol- 5-yl)methyl]-1,3-benzothiazol-2-amine (1.00 g, 3.8 mmol, 68 %) as a white solid. LCMS (ESI) m/z: 261.0 [M+H]+. Step 2: Preparation of N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2-yl)benzamide
Figure imgf000184_0001
To a solution of N-((1H-imidazol-4-yl)methyl)-6-methoxybenzo[d]thiazol-2-amine (200 mg, 0.76 mmol) in N,N-dimethylformamide (5 mL) was added benzoic acid (185 mg, 1.5 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (577 mg, 1.5 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. Water (20 mL) was added, then the reaction was extracted using 2x 20 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified prep-HPLC to give N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2- yl)benzamide (80.0 mg, 0.22 mmol, 34 %) as white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.57 – 8.45 (m, 2H), 8.04 (dd, J = 8.8, 5.5 Hz, 1H), 7.59 (dd, J = 10.1, 2.6 Hz, 2H), 7.43 – 7.25 (m, 1H), 7.25 – 7.07 (m, 6H), 5.59 (s, 2H), 2.83 – 2.52 (m, 4H), 2.13 – 1.63 (m, 2H); LCMS (ESI) m/z: 365.0 [M+H]+. Example 145. Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]benzamide
Figure imgf000184_0002
Step 1: Preparation of 6-ethyl-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000184_0003
To a solution of 1H-imidazole-4-carbaldehyde (0.799 g, 8.3 mmol), 6-ethylbenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (0.308 g, 8.3 mmol) in methanol (50 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-[(1H-imidazol-5- yl)methyl]-1,3-benzothiazol-2-amine (1.20 g, 4.6 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 259.0 [M+H]+. Step 2: Preparation of 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]benzamide
Figure imgf000184_0004
To a solution of N-((1H-imidazol-4-yl)methyl)-6-ethylbenzo[d]thiazol-2-amine (200 mg, 0.76 mmol) in N,N-dimethylformamide (5 mL) was added 4-cyanobenzoic acid (223 mg, 1.5 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (577 mg, 1.5 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. Water (20 mL) was added, then the reaction was extracted using 2x 20 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified prep-HPLC to give 4-cyano-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]benzamide (80.0 mg, 0.21 mmol, 34 %) as white solid.1H NMR (400 MHz, Dimethylsulfoxide- d6) δ 8.57 – 8.45 (m, 2H), 8.04 (dd, J = 8.8, 5.5 Hz, 1H), 7.59 (dd, J = 10.1, 2.6 Hz, 2H), 7.43 – 7.25 (m, 1H), 7.25 – 7.07 (m, 6H), 5.59 (s, 2H), 2.83 – 2.52 (m, 4H), 2.13 – 1.63 (m, 2H); LCMS (ESI) m/z: 387.0 [M+H]+. Example 146. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3- yl)methyl]acetamide
Figure imgf000185_0001
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000185_0002
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-2-phenyl-N-[(pyridin-3-yl)methyl]acetamide
Figure imgf000185_0003
In a reaction vial, 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (260 mg, 0.97 mmol) and 2-phenylacetic acid (158 mg, 1.2 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (550 mg, 1.5 mmol) and N,N- diisopropylethylamine (374 mg, 2.9 mmol) were dissolved in dichloromethane (5 mL). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. Water (20 mL) was added, then the reaction was extracted using 2x 20 mL washes of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by flash chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane to give N- (6-ethylbenzo[d]thiazol-2-yl)-2-phenyl-N-(pyridin-3-ylmethyl)acetamide (47.0 mg, 0.12 mmol, 13 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.57 (s, 1H), 8.48-8.57 (m, 1H), 7.81–7.81 (d, J = 1.2 Hz, 2H), 7.67–7.66 (m, 2H), 7.20–7.37 (m, 7H), 5.71 (s, 2H), 4.10 (s, 2H), 2.69–2.71 (m, 2H), 1.20– 1.23 (t, 3H); LCMS (ESI) m/z: 388.0 [M+H]+. Example 147. Preparation of (1r,3s)-N-(6-ethyl-1,3-benzothiazol-2-yl)-3-methyl-N-[(pyridin-3- yl)methyl]cyclobutane-1-carboxamide
Figure imgf000186_0001
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000186_0002
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of (1r,3s)-N-(6-ethyl-1,3-benzothiazol-2-yl)-3-methyl-N-[(pyridin-3- yl)methyl]cyclobutane-1-carboxamide
Figure imgf000186_0003
To a solution of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (120 mg, 0.45 mmol) in dichloromethane (15 mL) were added 3-methylcyclobutanecarboxylic acid (76.4 mg, 0.67 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (255 mg, 0.67 mmol) and triethylamine (136 mg, 1.4 mmol). The reaction was stirred at 10 °C for 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was further purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Racemic product N-(6- ethylbenzo[d]thiazol-2-yl)-3-methyl-N-(pyridin-3-ylmethyl)cyclobutanecarboxamide (80.0 mg, 0.22 mmol, 49 %) was afforded as a white solid.80 mg of this product was sent to chiral separation (the crude sample was dissolved in 35 mL methanol. AD 20*250 mm column. The mobile phase was 0.2% methanol/ammonia) to afford two isomers. Product (1r,3s)-N-(6-ethylbenzo[d]thiazol-2-yl)-3-methyl-N- (pyridin-3-ylmethyl)cyclobutanecarboxamide (4.60 mg, 0.010 mmol, 2.2 %) as white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.49 (d, J = 17.8 Hz, 2H), 7.82 (s, 1H), 7.59 (dd, J = 20.2, 7.6 Hz, 2H), 7.33 (s, 1H), 7.26 (d, J = 8.2 Hz, 1H), 5.47 (s, 2H), 3.84 – 3.57 (m, 1H), 2.71 (dd, J = 14.7, 7.2 Hz, 2H), 2.44 – 2.27 (m, 3H), 1.75 (s, 2H), 1.22 (t, J = 7.3 Hz, 3H), 1.09 (d, J = 5.9 Hz, 3H); LCMS (ESI) m/z: 366.0 [M+H]+. Example 148. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-2- yl)methyl]cyclohexanecarboxamide
Figure imgf000187_0001
Step 1: Preparation of 6-fluoro-N-[(1H-imidazol-2-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000187_0002
In a reaction vial, a solution of 6-fluoro-1,3-benzothiazol-2-amine (200 mg, 1.2 mmol) and 1H- imidazole-2-carbaldehyde (113 mg, 1.2 mmol) in dry toluene (12 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry ethyl alcohol (12 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate and concentrated under reduced pressure. The product was purified through flash column chromatography on a 24 gram silica gel column using a gradient of 0- 100% ethyl acetate in hexanes. Product 6-fluoro-N-[(1H-imidazol-2-yl)methyl]-1,3-benzothiazol-2-amine (198 mg, 0.80 mmol, 68 %) was afforded as a brown solid.1H NMR (300 MHz, Methanol-d4) δ 7.42 (tdd, J = 8.7, 3.8, 1.4 Hz, 2H), 7.16 – 6.90 (m, 3H), 4.70 (d, J = 1.4 Hz, 2H); LCMS (ESI) m/z: 249.0 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-2- yl)methyl]cyclohexanecarboxamide
Figure imgf000187_0003
In a reaction vial, 6-fluoro-N-[(1H-imidazol-2-yl)methyl]-1,3-benzothiazol-2-amine (199 mg, 0.80 mmol) and triethylamine (0.17 mL, 1.2 mmol) were dissolved in tetrahydrofuran (1.6 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (174 mg, 1.2 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-2- yl)methyl]cyclohexanecarboxamide (193 mg, 0.54 mmol, 68 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 7.83 (dd, J = 8.8, 4.6 Hz, 1H), 7.51 (dd, J = 8.0, 2.6 Hz, 1H), 7.21 (td, J = 8.9, 2.6 Hz, 1H), 7.01 (s, 2H), 5.47 (s, 2H), 3.35 (t, J = 10.9 Hz, 1H), 1.83 (d, J = 12.3 Hz, 5H), 1.69 – 1.39 (m, 4H), 1.38 – 1.19 (m, 1H); LCMS (ESI) m/z: 359.2 [M+H]+. Example 149. Preparation of 3-[(1H-imidazol-5-yl)methyl]-3-(6-methoxy-1,3-benzothiazol-2-yl)-1- phenylurea
Figure imgf000188_0001
Step 1: Preparation of 6-methoxy-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000188_0002
To a solution of 1H-imidazole-4-carbaldehyde (0.799 g, 8.3 mmol), 6-methoxybenzo[d]thiazol-2- amine (1.00 g, 5.6 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (0.308 g, 8.3 mmol) in methanol (50 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-methoxy-N-[(1H-imidazol- 5-yl)methyl]-1,3-benzothiazol-2-amine (1.00 g, 3.8 mmol, 68 %) as a white solid. LCMS (ESI) m/z: 261.0 [M+H]+. Step 2: Preparation of 3-[(1H-imidazol-5-yl)methyl]-3-(6-methoxy-1,3-benzothiazol-2-yl)-1-phenylurea
Figure imgf000188_0003
To a solution of triphosgene (113 mg, 0.38 mmol) in dichloromethane (10 mL) was added a solution of N-((1H-imidazol-5-yl)methyl)-6-methoxybenzo[d]thiazol-2-amine (200 mg, 0.77 mmol) and pyridine (182 mg, 2.3 mmol) in dichloromethane (5 mL) at room temperature under argon. The reaction was stirred at room temperature for 2 hours. Then a solution of aniline (113 mg, 0.77 mmol) and pyridine (182 mg, 2.3 mmol) in dichloromethane (5 mL) was added at room temperature. The reaction was stirred at room temperature for 4 hours. After completed, water (100 mL) was added, and reaction was extracted with 3x 50 mL washes of dichloromethane. The organic layers were pooled, dried over sodium sulfate, filtered and concentrated. Crude product was purified by silica gel column using a gradient of 0-50% ethyl acetate in petroleum ether. Product 3-[(1H-imidazol-5-yl)methyl]-3-(6-methoxy-1,3-benzothiazol-2-yl)-1- phenylurea (51.0 mg, 0.13 mmol, 17 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 12.29 (s, 1H), 11.15 (s, 1H), 7.83 (s, 1H), 7.67–7.69 (d, J = 8.8 Hz, 1H), 7.50–7.59 (m, 1H), 7.50 (s, 1H), 7.37–7.41 (t, 2H), 7.26 (s, 1H), 7.10–7.13 (t, 1H), 5.39 (s, 2H), 3.80 (s, 3H); LCMS (ESI) m/z: 380.0 [M+H]+. Example 150. Preparation of tert-butyl 4-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3- yl)methyl]carbamoyl]piperazine-1-carboxylate
Figure imgf000189_0001
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000189_0002
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of tert-butyl 4-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3- yl)methyl]carbamoyl]piperazine-1-carboxylate
Figure imgf000189_0003
To a solution of triphosgene (113 mg, 0.38 mmol) in dichloromethane (10 mL) was added a solution of N-((1H-imidazol-5-yl)methyl)-6-ethylbenzo[d]thiazol-2-amine (200 mg, 0.77 mmol) and pyridine (182 mg, 2.3 mmol) in dichloromethane (5 mL) at room temperature under argon. The reaction was stirred at room temperature for 2 hours. Then a solution of tert-butyl piperazine-1-carboxylate (143 mg, 0.77 mmol) and pyridine (182 mg, 2.3 mmol) in dichloromethane (5 mL) was added at room temperature. The reaction was stirred at room temperature for 4 hours. The product was indicated present via UPLC analysis. Water (100 mL) was added, and reaction was extracted with 3x 50 mL washes of dichloromethane. The organic layers were pooled, dried over sodium sulfate, filtered and concentrated. Crude product was purified by silica gel column using a gradient of 0-50% ethyl acetate in petroleum ether. Product tert-butyl 4-((6-ethylbenzo[d]thiazol-2-yl)(pyridin-3-ylmethyl)carbamoyl)piperazine-1- carboxylate (36.0 mg, 0.075 mmol, 12 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.58–8.58 (d, J = 1.6 Hz, 1H), 8.43–8.45 (m, 1H), 7.75–7.77 (d, J = 8.0 Hz, 1H), 7.69 (s, 1H), 7.56–7.58 (d, J = 8.4 Hz, 1H), 7.33–7.35 (t, 1H), 7.20–7.23 (m, 1H), 5.13 (s, 2H), 3.36–3.37 (m, 8H), 2.65–2.67 (m, 2H), 1.40 (s, 9H), 1.17–1.21 (t, 3H); LCMS (ESI) m/z: 482.0 [M+H]+. Example 151. Preparation of 4-acetyl-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]piperazine-1-carboxamide
Figure imgf000190_0001
Step 1: Preparation of 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000190_0002
To a solution of nicotinaldehyde (2.70 g, 25 mmol), 6-ethylbenzo[d]thiazol-2-amine (3.00 g, 17 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.89 g, 50 mmol) in ethanol (20 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (2.50 g, 9.3 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of 4-acetyl-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperazine-1- carboxamide
Figure imgf000190_0003
To a solution of triphosgene (113 mg, 0.38 mmol) in dichloromethane (10 mL) was added a solution of N-((1H-imidazol-5-yl)methyl)-6-ethylbenzo[d]thiazol-2-amine (200 mg, 0.77 mmol) and pyridine (182 mg, 2.3 mmol) in dichloromethane (5 mL) at room temperature under argon. The reaction was stirred at room temperature for 2 hours. Then a solution of 1-(piperazin-1-yl)ethanone (71.0 mg, 0.77 mmol) and pyridine (182 mg, 2.3 mmol) in dichloromethane (5 mL) was added at room temperature. The resulting mixture was stirred at room temperature for 4 hours. The product was indicated present via UPLC analysis. Water (100 mL) was added, and reaction was extracted with 3x 50 mL washes of dichloromethane. The organic layers were pooled, dried over sodium sulfate, filtered and concentrated. Crude product was purified by silica gel column using a gradient of 0-50% ethyl acetate in petroleum ether. Product 4-acetyl-N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)piperazine-1-carboxamide (96.0 mg, 0.23 mmol, 29 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.59 (d, J = 1.6 Hz, 1H), 8.43–8.45 (m, 1H), 7.75–7.78 (m, 1H), 7.69 (d, J = 1.6 Hz, 1H), 7.23–7.36 (m, 1H), 7.21–7.23 (m, 1H), 5.14 (s, 2H), 3.34–3.47 (m, 8H), 2.63–2.69 (m, 2H), 2.00 (s, 3H), 1.17–1.23 (t, 3H); LCMS (ESI) m/z: 424.0 [M+H]+. Example 152. Preparation of N-(1,3-benzothiazol-2-yl)-4-cyano-N-[(1H-imidazol-5- yl)methyl]benzamide
Figure imgf000191_0001
Step 1: Preparation of N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine
Figure imgf000191_0002
To a solution of benzo[d]thiazol-2-amine (1.10 g, 7.4 mmol), 1-trityl-1H-imidazole-4-carbaldehyde (2.50 g, 7.4 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.40 g, 37 mmol) in ethanol (7.4 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-((1-trityl-1H-imidazol-4- yl)methyl)benzo[d]thiazol-2-amine (2.20 g, 4.7 mmol, 63 %) as a yellow solid. LCMS (ESI) m/z: 473.1[M+H]+. Step 2: Preparation of N-(benzo[d]thiazol-2-yl)-4-cyano-N-((1-trityl-1H-imidazol-5-yl)methyl)benzamide
Figure imgf000191_0003
A mixture of N-((1-trityl-1H-imidazol-5-yl)methyl)benzo[d]thiazol-2-amine (400 mg, 0.85 mmol), 4- cyanobenzoic acid (125 mg, 0.85 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (485 mg, 1.3 mmol) and N,N- diisopropylethylamine (219 mg, 1.7 mmol) in tetrahydrofuran (20 mL) was stirred at 70 °C for 16 hours. The product was indicated present via UPLC analysis. The mixture was quenched with ice water, then extracted with 2x 15 mL portions of ethyl acetate. The organic layers were pooled, washed with 10 mL brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by flash chromatography through silica gel using a gradient of 0-35% ethyl acetate in petroleum ether. Product N-(benzo[d]thiazol-2-yl)-4-cyano-N-((1-trityl-1H-imidazol-5-yl)methyl)benzamide (250 mg, 0.42 mmol, 49 %) was afforded as a light yellow solid. Step 3: Preparation of N-(1,3-benzothiazol-2-yl)-4-cyano-N-[(1H-imidazol-5-yl)methyl]benzamide
Figure imgf000192_0001
To a solution of N-(benzo[d]thiazol-2-yl)-4-cyano-N-((1-trityl-1H-imidazol-5-yl)methyl)benzamide (100 mg, 0.17 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (2 mL). The reaction was stirred at room temperature for 3 hours. The product was indicated present via UPLC analysis. The reaction was concentrated under reduced pressure and quenched with a solution of 7 M ammonia in methanol until pH 8. The mixture was concentrated and purified directly by prep-HPLC (the crude samples were dissolved in methanol before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous ammonium bicarbonate). Product N-((1H-imidazol-5- yl)methyl)-N-(benzo[d]thiazol-2-yl)-4-cyanobenzamide (10.6 mg, 0.030 mmol, 18 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 11.93 (s, 1H), 8.04 – 7.93 (m, 5H), 7.83 (d, J = 7.9 Hz, 1H), 7.54 – 7.43 (m, 2H), 7.40 – 7.33 (m, 1H), 6.97 (s, 1H), 5.25 (s, 2H); LCMS (ESI) m/z: 360.1 [M+H]+. Example 153. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-4-methyl-N-[(pyridin-3- yl)methyl]cyclohexane-1-carboxamide
Figure imgf000192_0002
Step 1: Preparation of 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000192_0003
In a round bottom flask equipped with a stir bar, a solution of pyridine-3-carbaldehyde (0.63 mL, 6.7 mmol) and 6-ethyl-1,3-benzothiazol-2-amine (1.20 g, 6.7 mmol) in dry toluene (34 mL) was stirred at 105°C with activated 4 Å molecular sieves under nitrogen overnight. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (1.27 g, 34 mmol) in dry ethyl alcohol (34 mL). The reaction mixture was stirred at 70 °C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with ethyl acetate. The filtrate was concentrated then diluted with dichloromethane and filtered again. The filtrate was purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane through a 40 g column of silica gel. Product 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3- benzothiazol-2-amine (0.915 g, 3.4 mmol, 51 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.72 – 8.50 (m, 2H), 7.76 (dddd, J = 7.9, 2.4, 1.7, 0.8 Hz, 1H), 7.50 – 7.37 (m, 2H), 7.35 – 7.22 (m, 1H), 7.14 (dt, J = 8.3, 1.2 Hz, 1H), 6.24 (s, 1H), 4.68 (s, 2H), 2.70 (q, J = 7.6 Hz, 2H), 1.26 (td, J = 7.6, 0.7 Hz, 3H); LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of 4-methylcyclohexane-1-carbonyl chloride
Figure imgf000193_0001
In a reaction vial, 4-methylcyclohexane-1-carboxylic acid (300 mg, 2.1 mmol) was dissolved in dichloromethane (11 mL) and cooled to 0 °C. To the reaction was added oxalyl chloride (0.89 mL, 11 mmol) slowly. A few drops of N,N-dimethylformamide were added as a catalyst and the reaction bubbled. The reaction was then heated to 40 °C and stirred overnight. The product was indicated present via crude NMR analysis. The reaction was cooled to room temperature and concentrated under reduced pressure. Crude product was indicated pure enough to continue to next step via NMR and assumed 100% yield. Product 4-methylcyclohexane-1-carbonyl chloride (337 mg, 2.1 mmol, 100 %) was afforded as a clear oil. 1H NMR (300 MHz, Chloroform-d) δ 11.84 (s, 1H), 10.99 (s, 1H), 3.89 (s, 3H), 3.20 – 2.77 (m, 1H), 2.19 – 1.93 (m, 1H), 1.80 – 1.06 (m, 3H), 1.02 – 0.71 (m, 2H). Step 3: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-4-methyl-N-[(pyridin-3-yl)methyl]cyclohexane-1- carboxamide
Figure imgf000193_0002
In a reaction vial, 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (400 mg, 1.5 mmol) was dissolved in tetrahydrofuran (7.4 mL) and cooled to 0 °C. To the solution was added triethylamine (0.31 mL, 2.2 mmol) then 4-methylcyclohexane-1-carbonyl chloride (337 mg, 2.1 mmol) was added slowly. The reaction was stirred from 0 °C to room temperature for 1.5 hours. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane and washed with water (3x). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Crude product was purified via flash column chromatography eluting with 0-100% hexanes/ethyl acetate through 24 g of silica gel. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-4-methyl-N-[(pyridin-3-yl)methyl]cyclohexane-1- carboxamide (206 mg, 0.52 mmol, 35 %) was afforded as a colorless oil.1H NMR (300 MHz, Chloroform- d) δ 8.72 – 8.48 (m, 2H), 7.81 – 7.48 (m, 3H), 7.37 – 7.13 (m, 2H), 5.61 (s, 2H), 2.77 (q, J = 7.6 Hz, 3H), 2.10 – 1.37 (m, 8H), 0.95 (dd, J = 36.9, 6.8 Hz, 3H); LCMS (ESI) m/z: 394.3 [M+H]+. Example 154. Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(4-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000194_0001
Step 1: Preparation of 6-methoxy-N-[(4-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000194_0002
In a reaction vial, a solution of 4-methoxypyridine-3-carbaldehyde (150 mg, 1.1 mmol) and 6- methoxy-1,3-benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (95.7 mg, 2.5 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-methoxy-N-[(4-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (255 mg, 0.85 mmol, 77 %) was afforded as a pink solid.1H NMR (300 MHz, Methanol-d4) δ 8.37 (q, J = 2.1, 1.5 Hz, 2H), 7.33 (dd, J = 8.8, 1.2 Hz, 1H), 7.20 (dt, J = 2.8, 1.3 Hz, 1H), 7.10 (dd, J = 5.9, 1.2 Hz, 1H), 6.87 (ddd, J = 8.8, 2.7, 1.2 Hz, 1H), 4.62 (s, 2H), 3.99 (d, J = 1.2 Hz, 3H), 3.80 (d, J = 1.2 Hz, 3H); LCMS (ESI) m/z: 302.1 [M+H]+. Step 2: Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(4-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000194_0003
In a reaction vial, 6-methoxy-N-[(4-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (255 mg, 0.85 mmol) and triethylamine (0.17 mL, 1.3 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (368 mg, 2.6 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-50% ethyl acetate in hexanes. Product N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(4-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide (103 mg, 0.25 mmol, 30 %) was afforded as a white solid. 1H NMR (300 MHz, Chloroform-d) δ 8.43 (d, J = 5.7 Hz, 1H), 8.25 (s, 1H), 7.64 (d, J = 8.8 Hz, 1H), 6.99 (dd, J = 8.9, 2.5 Hz, 1H), 6.83 (d, J = 5.7 Hz, 1H), 5.53 (s, 2H), 3.95 (s, 3H), 3.86 (s, 3H), 2.78 – 2.64 (m, 1H), 1.80 (d, J = 12.8 Hz, 4H), 1.63 (q, J = 12.5, 12.0 Hz, 4H), 1.22 (dd, J = 17.0, 9.8 Hz, 3H); LCMS (ESI) m/z: 412.2 [M+H]+. Example 155. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(5-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000195_0001
Step 1: Preparation of 6-ethyl-N-[(5-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000195_0002
In a reaction vial, a solution of 5-methoxypyridine-3-carbaldehyde (153 mg, 1.1 mmol) and 6- ethyl-1,3-benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-ethyl-N-[(5-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (270 mg, 0.90 mmol, 81 %) was afforded as a brown solid.1H NMR (300 MHz, Methanol-d4) δ 8.23 – 8.09 (m, 2H), 7.49 – 7.37 (m, 2H), 7.35 (dd, J = 8.3, 1.1 Hz, 1H), 7.11 (dt, J = 8.3, 1.4 Hz, 1H), 4.66 (s, 2H), 3.86 (d, J = 1.4 Hz, 2H), 2.66 (q, J = 7.6 Hz, 2H), 1.23 (td, J = 7.6, 1.2 Hz, 3H); LCMS (ESI) m/z: 300.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(5-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000195_0003
In a reaction vial, 6-ethyl-N-[(5-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (271 mg, 0.90 mmol) and triethylamine (0.19 mL, 1.4 mmol) were dissolved in dimethylformamide (1.1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (197 mg, 1.4 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-25% ethyl acetate in hexanes. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(5-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide (298 mg, 0.73 mmol, 81 %) was afforded as a white solid. 1H NMR (300 MHz, Chloroform-d) δ 8.23 (d, J = 2.8 Hz, 1H), 8.20 (s, 1H), 7.68 (d, J = 8.3 Hz, 1H), 7.63 (d, J = 1.0 Hz, 1H), 7.26 (dd, J = 8.3, 1.7 Hz, 1H), 7.10 (t, J = 2.3 Hz, 1H), 5.59 (s, 2H), 3.78 (d, J = 0.7 Hz, 3H), 2.76 (q, J = 7.6 Hz, 3H), 1.69 (dq, J = 34.0, 12.4, 11.7 Hz, 7H), 1.41 – 1.02 (m, 5H); LCMS (ESI) m/z: 410.3 [M+H]+. Example 156. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(6-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000196_0001
Step 1: Preparation of 6-ethyl-N-[(6-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000196_0002
In a reaction vial, a solution of 6-methoxypyridine-3-carbaldehyde (153 mg, 1.1 mmol) and 6- ethyl-1,3-benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-ethyl-N-[(6-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (331 mg, 1.1 mmol, 99 %) was afforded as a white solid.1H NMR (300 MHz, Methanol-d4) δ 8.16 (dt, J = 2.6, 0.9 Hz, 1H), 7.76 – 7.59 (m, 1H), 7.42 (d, J = 1.7 Hz, 1H), 7.35 (d, J = 8.2 Hz, 1H), 7.11 (dd, J = 8.3, 1.7 Hz, 1H), 6.79 (dt, J = 8.6, 0.9 Hz, 1H), 4.55 (s, 2H), 3.31 (p, J = 1.6 Hz, 3H), 2.66 (q, J = 7.6 Hz, 2H), 1.23 (td, J = 7.6, 0.9 Hz, 3H); LCMS (ESI) m/z: 300.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(6-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000197_0001
In a reaction vial, 6-ethyl-N-[(6-methoxypyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (332 mg, 1.1 mmol) and triethylamine (0.23 mL, 1.7 mmol) were dissolved in dimethylformamide (1.4 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (241 mg, 1.7 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of dichloromethane were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-15% ethyl acetate in hexanes. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(6-methoxypyridin-3- yl)methyl]cyclohexanecarboxamide (173 mg, 0.42 mmol, 38 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.29 – 7.99 (m, 1H), 7.70 (d, J = 8.3 Hz, 1H), 7.62 (dd, J = 1.7, 0.7 Hz, 1H), 7.53 (dd, J = 8.6, 2.6 Hz, 1H), 7.31 – 7.12 (m, 1H), 6.68 (dd, J = 8.6, 0.7 Hz, 1H), 5.51 (s, 2H), 3.91 (s, 3H), 2.76 (q, J = 7.6 Hz, 3H), 2.04 – 1.49 (m, 8H), 1.28 (q, J = 6.7, 5.8 Hz, 5H); LCMS (ESI) m/z: 410.3 [M+H]+. Example 157. Preparation of 1-cyclopropanecarbonyl-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin- 3-yl)methyl]piperidine-4-carboxamide
Figure imgf000197_0002
Preparation of 1-cyclopropanecarbonyl-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]piperidine-4-carboxamide
Figure imgf000197_0003
To a solution of N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)piperidine-4-carboxamide (50.0 mg, 0.13 mmol) in dichloromethane (3 mL) was added triethylamine (26.0 mg, 0.26 mmol) and cyclopropanecarbonyl chloride (16.0 mg, 0.18 mmol). The reaction was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The reaction was diluted with water (3 mL), then extracted with 3x 15 mL portions of dichloromethane. The organic layers were separated, pooled, filtered over sodium sulfate, and concentrated under reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile/0.1% ammonium bicarbonate). Product 1-(cyclopropanecarbonyl)-N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)piperidine-4- carboxamide (39.4 mg, 0.088 mmol, 68 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.58 (s, 1H), 8.49 (d, J = 4.7 Hz, 1H), 7.82 (s, 1H), 7.66 – 7.62 (m, 2H), 7.36 (dd, J = 7.9, 4.8 Hz, 1H), 7.28 (d, J = 8.4, Hz, 1H), 5.70 (s, 2H), 4.37 – 4.24 (m, 2H), 3.37 – 3.24 (m, 1H), 3.15 – 3.07 (m, 1H), 2.74 – 2.57 (m, 3H), 2.00 - 1.94 (m, 1H), 1.82 - 1.36 (m, 4H), 1.22 (t, J = 7.6 Hz, 3H), 0.70 (s, 4H); LCMS (ESI) m/z: 449.3 [M+H]+. Example 158. Preparation of methyl 4-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000198_0001
Preparation of methyl 4-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3-yl)methyl]carbamoyl]piperidine-1- carboxylate
Figure imgf000198_0002
To a solution of N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)piperidine-4-carboxamide (50.0 mg, 0.13 mmol) in dichloromethane (3 mL) was added triethylamine (26.0 mg, 0.26 mmol) and methyl carbonochloridate (15.0 mg, 0.16 mmol). The reaction was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The reaction was diluted with water (3 mL), then extracted with 3x 15 mL portions of dichloromethane. The organic layers were separated, pooled, filtered over sodium sulfate, and concentrated under reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile/0.1% ammonium bicarbonate). Product methyl 4-((6-ethylbenzo[d]thiazol-2-yl)(pyridin-3-ylmethyl)carbamoyl)piperidine-1-carboxylate (29.4 mg, 0.067 mmol, 51 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.58 (d, J = 1.7 Hz, 1H), 8.48 (d, J = 3.5 Hz, 1H), 7.81 (s, 1H), 7.66 – 762 (m, 2H), 7.36 (dd, J = 7.8, 4.7 Hz, 1H), 7.28 (d, J = 8.3 Hz, 1H), 5.69 (s, 2H), 3.98 – 3.94 (m, 2H), 3.59 (s, 3H), 3.27 – 3.12 (m, 1H), 2.87 – 2.79 (m, 2H), 2.71 (q, J = 7.5 Hz, 2H), 1.70 – 1.67 (m, 2H), 1.58 – 1.21 (m, 2H), 1.22 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 439.2 [M+H]+. Example 159. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(2-methoxypyridin-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000199_0001
Step 1: Preparation of 6-ethyl-N-((2-methoxypyridin-4-yl)methyl)benzo[d]thiazol-2-amine
Figure imgf000199_0002
To a solution of 6-ethylbenzo[d]thiazol-2-amine (300 mg, 1.7 mmol), 2-methoxyisonicotinaldehyde (233 mg, 1.7 mmol) in toluene (40 mL) was added dried 4 Å molecular sieves. The mixture was heated to 120 °C and stirred for 16 hours. Then a solution of sodium borohydride (323 mg, 8.5 mmol) in methanol (30 mL) was added. The reaction was stirred at 120 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-50% ethyl acetate in dichloromethane to offer 6-ethyl-N-((2-methoxypyridin-4- yl)methyl)benzo[d]thiazol-2-amine (300 mg, 1.0 mmol, 59%) as a yellow solid. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(2-methoxypyridin-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000199_0003
To a solution of 6-ethyl-N-((2-methoxypyridin-4-yl)methyl)benzo[d]thiazol-2-amine (150 mg, 0.50 mmol) in N, N-dimethylformamide (5 mL) was added 60% w/w sodium hydride in oil (30.0 mg, 0.75 mmol) and cyclohexanecarbonyl chloride (95.0 mg, 0.65 mmol). The mixture was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The mixture was quenched with ice water, then extracted with ethyl acetate (2x 15 mL). The organic layers were pooled, washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (the crude samples were dissolved in methanol before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous ammonium bicarbonate). Product N-(6- ethylbenzo[d]thiazol-2-yl)-N-((2-methoxypyridin-4-yl)methyl)cyclohexanecarboxamide (50.0 mg, 0.12 mmol, 24 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.12 (d, J = 5.3 Hz, 1H), 7.82 (d, J = 1.7 Hz, 1H), 7.62 (d, J = 8.3 Hz, 1H), 7.27 (dd, J = 8.4, 1.8 Hz, 1H), 6.82 (dd, J = 5.3, 1.5 Hz, 1H), 6.59 (s, 1H), 5.62 (s, 2H), 3.81 (s, 3H), 2.71 (q, J = 7.6 Hz, 3H), 1.66 (t, J = 16.0 Hz, 5H), 1.43 (q, J = 11.5, 10.9 Hz, 2H), 1.21 (q, J = 9.5, 8.5 Hz, 6H); LCMS (ESI) m/z: 410.0 [M+H]+. Example 160. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000200_0001
Step 1: Preparation of 5-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine
Figure imgf000200_0002
To a solution of 5-fluorobenzo[d]thiazol-2-amine (50.0 mg, 0.30 mmol), 1-trityl-1H-imidazole-4- carbaldehyde (151 mg, 0.45 mmol) in toluene (8 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (45.0 mg, 1.2 mmol) in methanol (10 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was carried onto next step. Crude product 5-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (102 mg, 0.21 mmol, 69 %) was afforded as a yellow solid. LCMS (ESI) m/z: 491.0 [M+H]+. Step 2: Preparation of N-(5-Fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide
Figure imgf000200_0003
To a solution of 5-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (100 mg, 0.20 mmol) and triethylamine (0.085 mL, 0.61 mmol) in dichloromethane (8 mL) was added cyclohexanecarbonyl chloride (45.0 mg, 0.31 mmol). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. Water (10 mL) was added to the reaction, then the organic layer was extracted with 2x 15 mL portions of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by flash column chromatography through silica gel using a gradient of 0-3% methanol in dichloromethane. Product N-(5- fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4-yl)methyl)cyclohexanecarboxamide (78.0 mg, 0.13 mmol, 65 %) was afforded as a yellow solid. LCMS (ESI) m/z: 601.0 [M+H]+. Step 3: Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000201_0001
To a solution of N-(5-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide (78.0 mg, 0.13 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1.5 mL). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The mixture was concentrated and purified by prep-HPLC to give product N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]cyclohexanecarboxamide (27.2 mg, 0.076 mmol, 29 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 11.99 (s, 1H), 7.80- 7.96 (m, 1H), 7.65–7.62 (m, 1H), 7.58 (s, 1H), 7.22–7.17 (m, 1H), 7.08 (s, 1H), 5.46 (s, 2H), 1.83-1.66 (m, 5H), 1.49-1.20 (m, 6H); LCMS (ESI) m/z: 358.0 [M+H]+. Example 161. Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000201_0002
Step 1: Preparation of 4-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine
Figure imgf000202_0001
To a solution of 4-fluorobenzo[d]thiazol-2-amine (50.0 mg, 0.30 mmol), 1-trityl-1H-imidazole-4- carbaldehyde (151 mg, 0.45 mmol) in toluene (8 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (45.0 mg, 1.2 mmol) in methanol (10 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was carried onto next step. Crude product 4-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (102 mg, 0.21 mmol, 69 %) was afforded as a yellow solid. LCMS (ESI) m/z: 491.0 [M+H]+. Step 2: Preparation of N-(4-Fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide
Figure imgf000202_0002
To a solution of 4-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (100 mg, 0.20 mmol) and triethylamine (0.085 mL, 0.61 mmol) in dichloromethane (8 mL) was added cyclohexanecarbonyl chloride (45.0 mg, 0.31 mmol). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. Water (10 mL) was added to the reaction, then the organic layer was extracted with 2x 15 mL portions of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by flash column chromatography through silica gel using a gradient of 0-3% methanol in dichloromethane. Product N-(4- fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4-yl)methyl)cyclohexanecarboxamide (80.0 mg, 0.13 mmol, 65 %) was afforded as a yellow solid. LCMS (ESI) m/z: 601.0 [M+H]+. Step 3: Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000203_0001
To a solution of N-(4-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide (80.0 mg, 0.13 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1.5 mL). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The mixture was concentrated and purified by prep-HPLC to give product N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]cyclohexanecarboxamide (9.50 mg, 0.027 mmol, 14 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 11.99 (s, 1H), 7.80- 7.78 (m, 1H), 7.59 (s, 1H), 7.34-7.26 (m, 2H), 7.07 (s, 1H), 5.48 (s, 2H), 1.84-1.66 (m, 5H), 1.49-1.21 (m, 6H); LCMS (ESI) m/z: 358.0 [M+H]+. Example 162. Preparation of N-(7-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000203_0002
Step 1: Preparation of 7-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine
Figure imgf000203_0003
To a solution of 7-fluorobenzo[d]thiazol-2-amine (50.0 mg, 0.30 mmol), 1-trityl-1H-imidazole-4- carbaldehyde (151 mg, 0.45 mmol) in toluene (8 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (45.0 mg, 1.2 mmol) in methanol (10 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was carried onto next step. Crude product 7-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (102 mg, 0.21 mmol, 69 %) was afforded as a yellow solid. LCMS (ESI) m/z: 491.0 [M+H]+. Step 2: Preparation of N-(7-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide
Figure imgf000204_0001
To a solution of 7-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (100 mg, 0.20 mmol) and triethylamine (0.085 mL, 0.61 mmol) in dichloromethane (8 mL) was added cyclohexanecarbonyl chloride (45.0 mg, 0.31 mmol). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. Water (10 mL) was added to the reaction, then the organic layer was extracted with 2x 15 mL portions of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by flash column chromatography through silica gel using a gradient of 0-3% methanol in dichloromethane. Product N-(7- fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4-yl)methyl)cyclohexanecarboxamide (80.0 mg, 0.13 mmol, 65 %) was afforded as a yellow solid. LCMS (ESI) m/z: 601.0 [M+H]+. Step 3: Preparation of N-(7-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000204_0002
To a solution of N-(7-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide (80.0 mg, 0.13 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1.5 mL). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The mixture was concentrated and purified by prep-HPLC to give product N-(7-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]cyclohexanecarboxamide (14.1 mg, 0.039 mmol, 20 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 12.00 (s, 1H), 7.69- 7.67 (m, 1H), 7.59 (s, 1H), 7.50-7.45 (m, 1H), 7.22-7.17 (m, 1H), 7.10 (s, 1H), 5.47(s, 2H), 1.85-1.66 (m, 5H), 1.49-1.21 (m, 6H); LCMS (ESI) m/z: 358.0 [M+H]+. Example 163. Preparation of phenyl N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2- yl)carbamate
Figure imgf000205_0001
Step 1: Preparation of 6-methoxy-N-((1-trityl-1H-imidazol-5-yl)methyl)benzo[d]thiazol-2-amine
Figure imgf000205_0002
To a solution of 6-methoxybenzo[d]thiazol-2-amine (300 mg, 1.7 mmol), 1-trityl-1H-imidazole-4- carbaldehyde (563 mg, 1.7 mmol) in toluene (40 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (323 mg, 8.5 mmol) in methanol (30 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-30% ethyl acetate in dichloromethane. Product 6-methoxy-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (300 mg, 0.60 mmol, 35 %) was afforded as a white solid. Step 2: Preparation of phenyl (6-methoxybenzo[d]thiazol-2-yl)((1-trityl-1H-imidazol-5-yl)methyl)carbamate
Figure imgf000205_0003
To a solution of 6-methoxy-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (100 mg, 0.20 mmol) in N,N-dimethylformamide (5 mL) was added 60% w/w sodium hydride in oil (12.0 mg, 0.30 mmol) and phenyl carbonochloridate (41.0 mg, 0.26 mmol). The reaction was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. Water (10 mL) was added to the reaction, then the organic layer was extracted with 2x 15 mL portions of dichloromethane. The organic layers were pooled, washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by flash column chromatography through silica gel using a gradient of 0-25% ethyl acetate in petroleum ether. Product phenyl 6-methoxybenzo[d]thiazol-2- yl((1-trityl-1H-imidazol-5-yl)methyl)carbamate (100 mg, 0.16 mmol, 80 %) was afforded as a light yellow solid. Step 3: Preparation of phenyl N-[(1H-imidazol-4-yl)methyl]-N-(6-methoxy-1,3-benzothiazol-2-yl)carbamate
Figure imgf000206_0001
To a solution of (6-methoxybenzo[d]thiazol-2-yl)((1-trityl-1H-imidazol-5-yl)methyl)carbamate (100 mg, 0.16 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (2 mL), and the reaction was stirred at room temperature for 3 hours. The product was indicated present via UPLC analysis. The resulting mixture was concentrated and quenched with a solution of 7 M ammonia in methanol, until pH 8. The mixture was concentrated and purified directly by prep-HPLC (the crude samples were dissolved in methanol before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous ammonium bicarbonate). Product phenyl ((1H-imidazol-5-yl)methyl)(6- methoxybenzo[d]thiazol-2-yl)carbamate (29.9 mg, 0.079 mmol, 49 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 11.97 (s, 1H), 7.71 (d, J = 8.8 Hz, 1H), 7.61 – 7.54 (m, 2H), 7.47 (t, J = 7.9 Hz, 2H), 7.33 (t, J = 7.4 Hz, 1H), 7.26 (dd, J = 7.5, 1.5 Hz, 2H), 7.10 (s, 1H), 7.03 (dd, J = 8.9, 2.6 Hz, 1H), 5.47 (s, 2H), 3.80 (s, 3H); LCMS (ESI) m/z: 381.0 [M+H]+. Example 164. Preparation of 1-{bicyclo[1.1.1]pentane-1-carbonyl}-N-(6-ethyl-1,3-benzothiazol-2-yl)- N-[(pyridin-3-yl)methyl]piperidine-4-carboxamide
Figure imgf000206_0002
Preparation of 1-{bicyclo[1.1.1]pentane-1-carbonyl}-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]piperidine-4-carboxamide
Figure imgf000206_0003
A solution of N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)piperidine-4-carboxamide (50.0 mg, 0.13 mmol), bicyclo[1.1.1]pentane-1-carboxylic acid (18.0 mg, 0.16 mmol), 1H-benzo[d][1,2,3]triazol- 1-ol (35.0 mg, 0.26 mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (50.0 mg, 0.26 mmol) in N,N-dimethylformamide (5 mL) was stirred at room temperature for 16 hours. Water (20 mL) was added and the mixture was extracted with 2x 30 mL portions of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile/0.1% ammonium bicarbonate) to afford product 1-(bicyclo[1.1.1]pentane-1-carbonyl)-N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3- ylmethyl)piperidine-4-carboxamide (38.7 mg, 0.082 mmol, 62 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.58 (s, 1H), 8.49 (d, J = 4.7 Hz, 1H), 7.82 (s, 1H), 7.66 – 7.62 (m, 2H), 7.36 (dd, J = 7.9, 4.8 Hz, 1H), 7.28 (d, J = 8.4, Hz, 1H), 5.70 (s, 2H), 4.28 (d, J = 12.9 Hz, 1H), 4.13 (d, J = 13.9 Hz, 1H), 3.30 – 3.19 (m, 1H), 3.06 (t, J = 11.9 Hz, 1H), 2.71 (q, J = 7.5 Hz, 2H), 2.59 (t, J = 12.8 Hz, 1H), 2.44 (s, 1H), 2.22 – 1.96 (m, 6H), 1.71 (t, J = 16.1 Hz, 2H), 1.48 (dd, J = 30.6, 11.4 Hz, 2H), 1.22 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 475.2 [M+H]+. Example 165. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-1- propanoylpiperidine-4-carboxamide
Figure imgf000207_0001
Step 1: Preparation of 1-propionylpiperidine-4-carboxylic acid
Figure imgf000207_0002
To a solution of piperidine-4-carboxylic acid (300 mg, 2.3 mmol) in tetrahydrofuran (10 mL) was added N,N-diisopropylethylamine (593 mg, 4.6 mmol) and propionyl chloride (275 mg, 3.0 mmol). The resulting mixture was stirred at room temperature for 3 hours. The mixture was purified directly by reversed-phase chromatography (mobile phase was 15% acetonitrile/0.01% aqueous formic acid) to offer 1-propionylpiperidine-4-carboxylic acid (200 mg, 1.1 mmol, 48 %) as a white solid. Step 2: Preparation of N-(6-fluorobenzo[d]thiazol-2-yl)-1-propionyl-N-((1-trityl-1H-imidazol-5- yl)methyl)piperidine-4-carboxamide
Figure imgf000207_0003
A mixture of 6-fluoro-N-((1-trityl-1H-imidazol-5-yl)methyl)benzo[d]thiazol-2-amine (211 mg, 0.43 mmol), 1-propionylpiperidine-4-carboxylic acid (80.0 mg, 0.43 mmol), N-[(dimethylamino)-1H-1,2,3- triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (245 mg, 0.65 mmol), N,N-diisopropylethylamine (111 mg, 0.86 mmol) and tetrahydrofuran (15 mL) was stirred at 70ºC for 16 hours. The product was indicated present via UPLC analysis. The reaction was quenched with ice water, then extracted with ethyl acetate (2x 15 mL). The organic layers were pooled, washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by flash column chromatography through silica gel using a gradient of 0-25% ethyl acetate in petroleum ether. Product N-(6-fluorobenzo[d]thiazol-2-yl)-1-propionyl-N-((1-trityl-1H-imidazol-5- yl)methyl)piperidine-4-carboxamide (120 mg, 0.18 mmol, 43 %) was afforded as light yellow solid. Step 3: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-1- propanoylpiperidine-4-carboxamide
Figure imgf000208_0001
To a solution of N-(6-fluorobenzo[d]thiazol-2-yl)-1-propionyl-N-((1-trityl-1H-imidazol-5- yl)methyl)piperidine-4-carboxamide (120 mg, 0.18 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (2 mL). The reaction was stirred at room temperature for 3 hours, then concentrated under reduced pressure. The residue was quenched with a solution of 7 M ammonia in methanol until pH 8. The resulting mixture was concentrated and purified directly by prep-HPLC (the crude samples were dissolved in methanol before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-((1H-imidazol-5-yl)methyl)-N-(6- fluorobenzo[d]thiazol-2-yl)-1-propionylpiperidine-4-carboxamide (70.5 mg, 0.17 mmol, 94 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 12.03 (s, 1H), 7.91 – 7.78 (m, 2H), 7.59 (d, J = 1.2 Hz, 1H), 7.29 (td, J = 9.1, 2.7 Hz, 1H), 7.11 (s, 1H), 4.45 (d, J = 13.1 Hz, 1H), 3.91 (d, J = 13.6 Hz, 1H), 3.69 – 3.57 (m, 1H), 3.08 (t, J = 12.7 Hz, 1H), 2.68 – 2.56 (m, 1H), 2.40 – 2.20 (m, 2H), 1.84 (t, J = 12.4 Hz, 2H), 1.69 – 1.35 (m, 2H), 0.99 (t, J = 7.4 Hz, 3H); LCMS (ESI) m/z: 416.0 [M+H]+. Example 166. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-4- yl)methyl]propenamide
Figure imgf000208_0002
Step 1: Preparation of 5-fluoro-N-(pyridin-4-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000208_0003
To a solution of 5-fluorobenzo[d]thiazol-2-amine (340 mg, 2.0 mmol), isonicotinaldehyde (321 mg, 3.0 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (296 mg, 8.0 mmol) in methanol (10 mL) was added. The reaction was stirred at room temperature for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane. Product 5- fluoro-N-(pyridin-4-ylmethyl)benzo[d]thiazol-2-amine (500 mg, 1.9 mmol, 96 %) was afforded as a yellow solid. LCMS (ESI) m/z: 260.1 [M+H]+. Step 2: Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyridin-4-yl)methyl]propanamide
Figure imgf000209_0001
To a solution of 5-fluoro-N-(pyridin-4-ylmethyl)benzo[d]thiazol-2-amine (500 mg, 1.9 mmol) and pyridine (0.5 mL) in N,N-dimethylformamide (2 mL) and dichloromethane (5 mL), was added 3- phenoxypropanoic acid (379 mg, 2.3 mmol), 1,3-dicyclohexylcarbodiimide (597 mg, 2.9 mmol), and 1- hydroxybenzotriazole (391 mg, 2.9 mmol). The reaction mixture was stirred at room temperature overnight. The product was indicated present via UPLC analysis. The mixture was purified by prep-HPLC to give N-(5-fluorobenzo[d]thiazol-2-yl)-3-phenoxy-N-(pyridin-4-ylmethyl)propanamide (100 mg, 0.25 mmol, 13 %) as white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.53-8.54 (m, 2H), 8.04-8.07 (m, 1H), 7.56-7.59 (m, 1H), 7.23-7.29 (m, 5H), 6.89-6.95 (m, 3H), 5.69 (s, 2H), 4.30 (t, J = 6.0 Hz, 2H), 3.14 (t, J = 6.0 Hz, 2H); LCMS (ESI) m/z: 408.1 [M+H]+. Example 167. Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-3- phenoxypropanamide
Figure imgf000209_0002
Step 1: Preparation of 4-fluoro-N-[(1H-imidazol-4-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000209_0003
In a reaction vial, a solution of 1H-imidazole-4-carbaldehyde (113 mg, 1.2 mmol) and 4-fluoro-1,3- benzothiazol-2-amine (200 mg, 1.2 mmol) in dry toluene (12 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry ethyl alcohol (12 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-20% methanol in ethyl acetate. Product 4-fluoro-N-[(1H- imidazol-4-yl)methyl]-1,3-benzothiazol-2-amine (104 mg, 0.42 mmol, 36 %) was afforded as a white solid. 1H NMR (300 MHz, Methanol-d4) δ 7.66 (d, J = 1.1 Hz, 1H), 7.39 (dd, J = 6.3, 2.8 Hz, 1H), 7.10 (s, 1H), 7.08 – 6.96 (m, 2H), 4.60 (s, 2H); LCMS (ESI) m/z: 249.0 [M+H]+. Step 2: Preparation of 3-phenoxypropanoyl chloride
Figure imgf000210_0001
In a reaction vial, 3-phenoxypropanoic acid (500 mg, 3.0 mmol) was dissolved in dichloromethane (6 mL) and cooled to 0 °C while stirring. oxalyl chloride (1.26 mL, 15 mmol) was added dropwise, followed by a few drops of N,N-dimethylformamide. The reaction bubbled. The reaction was heated to 47°C and stir for 2 hours. The reaction was then cool to room temperature and concentrated under reduced pressure. Crude product was indicated pure enough to continue to next step via NMR. Product 3- phenoxypropanoyl chloride (549 mg, 3.0 mmol, 99 %) was afforded as a yellow oil.1H NMR (300 MHz, Chloroform-d) δ 7.35 – 7.27 (m, 1H), 6.99 (ddt, J = 7.7, 7.0, 1.0 Hz, 1H), 6.93 – 6.86 (m, 1H), 4.28 (t, J = 5.9 Hz, 1H), 3.35 (t, J = 5.9 Hz, 1H). Step 3: Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-3- phenoxypropanamide
Figure imgf000210_0002
In a reaction vial, 4-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine (104 mg, 0.42 mmol) and triethylamine (0.087 mL, 0.63 mmol) were dissolved in tetrahydrofuran (1 mL) and cooled to 0 °C.3-phenoxypropanoyl chloride (115 mg, 0.63 mmol) was added dropwise with stirring. The reaction was warmed to room temperature and stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of dichloromethane were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-3- phenoxypropanamide (54.1 mg, 0.14 mmol, 33 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 7.67 – 7.51 (m, 2H), 7.37 – 7.23 (m, 4H), 7.17 (ddd, J = 10.6, 8.1, 1.1 Hz, 1H), 7.01 – 6.84 (m, 3H), 5.52 (s, 2H), 4.45 (t, J = 6.4 Hz, 2H), 3.52 (t, J = 6.5 Hz, 2H); LCMS (ESI) m/z: 397.1 [M+H]+. Example 168. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-2- yl)methyl]cyclohexanecarboxamide
Figure imgf000211_0001
Step 1: Preparation of 6-ethyl-N-[(1H-imidazol-2-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000211_0002
In a reaction vial, a solution of 1H-imidazole-2-carbaldehyde (188 mg, 2.0 mmol) and 6-ethyl-1,3- benzothiazol-2-amine (350 mg, 2.0 mmol) in dry toluene (10 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (222 mg, 5.9 mmol) in dry ethyl alcohol (10 mL). The reaction mixture was stirred at 65°C for 1 hour and then cooled to room temperature. The reaction was filtered over a bed of Celite and washed with ethyl acetate (2x 20 mL). The filtrate was diluted with ethyl acetate and washed with sodium bicarbonate. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The filtrate was purified via flash column chromatography eluting with 0-100% hexanes/ethyl acetate, then 0-10% methanol in dichloromethane. Product 6-ethyl-N-[(1H-imidazol-2- yl)methyl]-1,3-benzothiazol-2-amine (323 mg, 1.3 mmol, 64 %) was afforded as a beige solid.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 11.94 (s, 1H), 8.32 (t, J = 5.2 Hz, 1H), 7.50 (d, J = 1.8 Hz, 1H), 7.32 (dd, J = 8.2, 1.3 Hz, 1H), 7.17 – 6.59 (m, 4H), 4.56 (dd, J = 5.4, 1.3 Hz, 2H), 3.34 (s, 1H), 1.18 (td, J = 7.5, 1.4 Hz, 4H); LCMS (ESI) m/z: 259.0 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-2- yl)methyl]cyclohexanecarboxamide
Figure imgf000211_0003
In a reaction vial, 6-ethyl-N-[(1H-imidazol-2-yl)methyl]-1,3-benzothiazol-2-amine (324 mg, 1.3 mmol) was dissolved in tetrahydrofuran (6 mL) and cooled to 0 °C. To the solution was added triethylamine (0.26 mL, 1.9 mmol) then cyclohexanecarbonyl chloride (0.25 mL, 1.9 mmol) was added, slowly. The reaction was stirred from 0 °C to room temperature for 3.5 hours. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane and washed with water (3x). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Crude product was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane through 24 g of silica gel. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-2- yl)methyl]cyclohexanecarboxamide (86.6 mg, 0.24 mmol, 19 %) was afforded as a pale pink solid.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 7.88 – 7.57 (m, 2H), 7.27 (d, J = 8.4 Hz, 1H), 7.04 (s, 1H), 6.80 (s, 1H), 5.56 (s, 2H), 1.90 – 1.54 (m, 6H), 1.52 – 1.01 (m, 8H); LCMS (ESI) m/z: 369.2 [M+H]+ Example 169. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-1-[2-(1-methylcyclopropyl)acetyl]-N- [(pyridin-3-yl)methyl]piperidine-4-carboxamide
Figure imgf000212_0001
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-1-[2-(1-methylcyclopropyl)acetyl]-N-[(pyridin-3- yl)methyl]piperidine-4-carboxamide
Figure imgf000212_0002
To a solution of N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)piperidine-4-carboxamide (50.0 mg, 0.13 mmol) in N,N-dimethylformamide (3 mL) was added 1-ethyl-3-(3- dimethylaminopropyl)carbodiimide hydrochloride (50.0 mg, 0.26 mmol), 1-hydroxybenzotriazole (35.0 mg, 0.26 mmol) and 2-(1-methylcyclopropyl)acetic acid (18.0 mg, 0.16 mmol). The reaction was stirred at room temperature for 20 hours. The product was indicated present via UPLC analysis. The mixture was diluted with water (3 mL), then extracted with dichloromethane (3x 15 mL). The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile/0.1% ammonium bicarbonate) to afford the desired compound N-(6- ethylbenzo[d]thiazol-2-yl)-1-(2-(1-methylcyclopropyl)acetyl)-N-(pyridin-3- ylmethyl)piperidine-4-carboxamide (17.5 mg, 0.040 mmol, 28 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.59 (s, 1H), 8.58 - 8.48 (m, 1H), 7.82 (s, 1H), 7.81 – 7.62 (m, 2H), 7.38 – 7.26 (m, 2H), 5.70 (s, 2H), 4.41 (d, J = 12.8 Hz, 1H), 3.87 (d, J = 13.2 Hz, 1H), 3.23 – 3.17 (m, 1H), 3.04 – 2.98 (m, 1H), 2.71 (q, J = 7.6 Hz, 2H), 2.58 – 2.53 (m, 1H), 2.42 – 2.18 (m, 2H), 1.71 – 1.43 (m, 4H), 1.21 (t, J = 7.6 Hz, 3H),1.06 (s, 3H), 0.41 – 0.09 (m, 4H); LCMS (ESI) m/z: 473.3 [M+H]+. Example 170. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-1-(3-hydroxy-3-methylbutanoyl)-N- [(pyridin-3-yl)methyl]piperidine-4-carboxamide
Figure imgf000212_0003
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-1-(3-hydroxy-3-methylbutanoyl)-N-[(pyridin-3- yl)methyl]piperidine-4-carboxamide
Figure imgf000213_0001
To a solution of N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)piperidine-4-carboxamide (50.0 mg, 0.13 mmol) in N,N-dimethylformamide (3 mL) was added 1-ethyl-3-(3- dimethylaminopropyl)carbodiimide hydrochloride (50.0 mg, 0.26 mmol), 1-hydroxybenzotriazole (35.0 mg, 0.26 mmol) and 3-hydroxy-3-methylbutanoic acid (19.0 mg, 0.16 mmol). The reaction was stirred at room temperature for 20 hours. The product was indicated present via UPLC analysis. The mixture was diluted with water (3 mL), then extracted with dichloromethane (3x 15 mL). The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile/0.1% ammonium bicarbonate) to afford the desired compound N-(6-ethylbenzo[d]thiazol-2-yl)-1-(3-hydroxy-3-methylbutanoyl)-N-(pyridin-3-ylmethyl)piperidine-4- carboxamide (8.40 mg, 0.020 mmol, 13 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.60 (s, 1H), 8.49 (d, J = 4.7 Hz, 1H), 7.82 (s, 4H), 7.70 – 7.57 (m, 2H), 7.39 – 7.26 (m, 2H), 5.71 (s, 2H), 4.86 (s, 1H), 4.45 (d, J = 13.0 Hz, 1H), 4.01 (d, J = 14.0 Hz, 1H), 3.34 – 3.19 (m, 2H), 3.03 – 3.00 (m, 2H), 2.71 (q, J = 7.5 Hz, 2H), 2.62 – 2.51 (m, 1H), 2.50 – 2.38 (m, 2H), 1.84 – 1.33 (m, 4H), 1.22 (t, J = 7.6 Hz, 3H), 1.19 (s, 6H); LCMS (ESI) m/z: 481.2 [M+H]+. Example 171. Preparation of cyclobutyl 4-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000213_0002
Preparation of cyclobutyl 4-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3-yl)methyl]carbamoyl]piperidine-1- carboxylate
Figure imgf000213_0003
To a solution of triphosgene (50.0 mg, 0.13 mmol) in tetrahydrofuran (5 mL) was added a solution of cyclobutanol (12.0 mg, 0.17 mmol) and triethylamine (39.0 mg, 0.39 mmol) in tetrahydrofuran (2 mL) at 0 °C under argon. The mixture was stirred at 0 °C for 2 hours. Then a solution of N-((1H-imidazol-5- yl)methyl)-6-methoxybenzo[d]thiazol-2-amine (50.0 mg, 0.13 mmol) was added at 0 °C. The resulting mixture was stirred at room temperature for 4 hours. The product was indicated present via UPLC analysis. Water (10 mL) was added, then the mixture was extracted with dichloromethane 3x 15 mL. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile/0.1% ammonium bicarbonate) to afford the desired compound cyclobutyl 4-((6-ethylbenzo[d]thiazol-2-yl)(pyridin-3- ylmethyl)carbamoyl)piperidine-1-carboxylate (5.10 mg, 0.010 mmol, 7.7 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.58 (s, 1H), 8.49 (d, J = 3.9 Hz, 1H), 7.82 (s, 1H), 7.66 – 7.62 (m, 2H), 7.38 – 7.27 (m, 2H), 5.69 (s, 2H), 4.82 (p, J = 7.3 Hz, 1H), 3.96 (d, J = 13.1 Hz, 2H), 3.20 – 3.09 (m, 1H), 2.90 – 2.68 (m, 4H), 2.25 – 2.20 (m, 2H), 2.01 – 1.95 (m, 2H), 1.70 – 1.66 (m, 3H).1.59 – 1.51 (m, 3H).1.34 – 1.16 (m, 3H); LCMS (ESI) m/z: 479.3 [M+H]+. Example 172. Preparation of tert-butyl 4-{N-[(pyridin-3-yl)methyl]6-ethyl-1,3-benzothiazole-2- amido}piperidine-1-carboxylate
Figure imgf000214_0001
Step 1: Preparation of tert-butyl 4-((pyridin-3-ylmethyl)amino)piperidine-1-carboxylate
Figure imgf000214_0002
In a reaction vial, a solution of tert-butyl 4-aminopiperidine-1-carboxylate (100 mg, 0.50 mmol) and nicotinaldehyde (69.0 mg, 0.65 mmol) in dry toluene (10 mL) was stirred at 110 °C with activated 4 Å molecular sieves under nitrogen for 16 hours. The reaction mixture was cooled to room temperature and then poured into a suspension of sodium borohydride (78.0 mg, 2.1 mmol) in dry methanol (20 mL). The reaction mixture was stirred at room temperature for 1 hour and then cooled to room temperature. The reaction was filtered over a bed of Celite and washed with ethyl acetate (2x 20 mL). The filtrate was diluted with ethyl acetate and washed with sodium bicarbonate. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The filtrate was purified via flash column chromatography eluting with 0-5% methanol in dichloromethane. Product tert-butyl 4-((pyridin-3- ylmethyl)amino)piperidine-1-carboxylate (100 mg, 0.34 mmol, 69 %) was afforded as a white solid. LCMS (ESI) m/z: 292.0 [M+H]+. Step 2: Preparation of tert-butyl 4-{N-[(pyridin-3-yl)methyl]6-ethyl-1,3-benzothiazole-2-amido}piperidine-1- carboxylate
Figure imgf000215_0001
A mixture of 6-ethylbenzo[d]thiazole-2-carboxylic acid (36.0 mg, 0.17 mmol), N,N- diisopropylethylamine (67.0 mg, 0.52 mmol) and N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (85.0 mg, 0.22 mmol) in dichloromethane (8 mL) was stirred at room temperature for 30 minutes, then tert-butyl 4-((pyridin-3- ylmethyl)amino)piperidine-1-carboxylate (50.0 mg, 0.17 mmol) was added. The reaction mixture was stirred at room temperature for 1.5 hours. The product was indicated present via UPLC analysis. The reaction mixture was diluted with 20 mL water and 20 mL dichloromethane. The organic layer was separated and concentrated under reduced pressure. Crude product was purified by prep-HPLC to give tert-butyl 4-{N-[(pyridin-3-yl)methyl]6-ethyl-1,3-benzothiazole-2-amido}piperidine-1-carboxylate (24.0 mg, 0.050 mmol, 29 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.57-8.42 (m, 2H), 8.09- 7.70 (m, 3H), 7.50-7.30 (m, 2H), 5.38 (s, 1H), 5.275.245 (m, 1H), 4.78 (s, 1H), 4.00 (s, 2H), 2.82 - 2.72 (m, 4H), 1.81 - 1.63 (m, 4H), 1.41 (s, 9H), 1.31 - 1.21 (m, 4H); LCMS (ESI) m/z: 481.0 [M+H]+. Example 173. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyrazin-2- yl)methyl]cyclohexanecarboxamide
Figure imgf000215_0002
Step 1: Preparation of 6-ethyl-N-[(pyrazin-2-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000215_0003
In a reaction vial, a solution of pyrazine-2-carbaldehyde (121 mg, 1.1 mmol) and 6-ethyl-1,3- benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-20% methanol in ethyl acetate. Product 6-ethyl-N-[(pyrazin-2- yl)methyl]-1,3-benzothiazol-2-amine (210 mg, 0.78 mmol, 70 %) was afforded as an orange solid.1H NMR (300 MHz, Methanol-d4) δ 8.69 (s, 1H), 8.60 (t, J = 2.0 Hz, 1H), 8.50 (d, J = 2.6 Hz, 1H), 7.44 (d, J = 1.7 Hz, 1H), 7.33 (d, J = 8.2 Hz, 1H), 7.10 (dd, J = 8.3, 1.7 Hz, 1H), 4.80 (s, 2H), 2.67 (q, J = 7.6 Hz, 2H), 1.23 (td, J = 7.6, 0.8 Hz, 3H); LCMS (ESI) m/z: 271.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyrazin-2-yl)methyl]cyclohexanecarboxamide
Figure imgf000216_0001
In a reaction vial, 6-ethyl-N-[(pyrazin-2-yl)methyl]-1,3-benzothiazol-2-amine (205 mg, 0.76 mmol) and N,N-diisopropylethylamine (0.39 mL, 2.3 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (165 mg, 1.1 mmol) was added dropwise with stirring. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-40% ethyl acetate in hexanes. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N- [(pyrazin-2-yl)methyl]cyclohexanecarboxamide (150 mg, 0.40 mmol, 52 %) was afforded as a cream solid. 1H NMR (300 MHz, Chloroform-d) δ 8.73 (d, J = 1.4 Hz, 1H), 8.61 – 8.39 (m, 2H), 7.66 (d, J = 8.3 Hz, 1H), 7.61 (d, J = 1.7 Hz, 1H), 7.32 – 7.13 (m, 1H), 5.68 (s, 2H), 2.95 (tt, J = 11.3, 3.4 Hz, 1H), 2.75 (q, J = 7.6 Hz, 2H), 1.85 (d, J = 12.4 Hz, 4H), 1.67 (d, J = 19.7 Hz, 3H), 1.28 (td, J = 7.0, 6.4, 3.2 Hz, 6H); LCMS (ESI) m/z: 381.2 [M+H]+. Example 174. Preparation of N-{6-methoxy-[1,3]thiazolo[4,5-b]pyridin-2-yl}-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000216_0002
Step 1: Preparation of 6-methoxy-N-[(pyridin-3-yl)methyl]-[1,3]thiazolo[4,5-b]pyridin-2-amine
Figure imgf000216_0003
In a reaction vial, a solution of 6-methoxy-[1,3]thiazolo[4,5-b]pyridin-2-amine (200 mg, 1.1 mmol) and pyridine-3-carbaldehyde (117 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (95.7 mg, 2.5 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The filtrate was concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-20% methanol in ethyl acetate. Product 6-methoxy-N-[(pyridin-3-yl)methyl]-[1,3]thiazolo[4,5- b]pyridin-2-amine (144 mg, 0.53 mmol, 48 %) was afforded as an orange solid.1H NMR (300 MHz, Methanol-d4) δ 8.60 (s, 1H), 8.45 (d, J = 4.9 Hz, 1H), 8.02 – 7.85 (m, 2H), 7.85 – 7.68 (m, 1H), 7.43 (dd, J = 7.4, 5.5 Hz, 1H), 4.73 (s, 2H), 3.86 (d, J = 1.4 Hz, 3H); LCMS (ESI) m/z: 273.0 [M+H]+. Step 2: Preparation of N-{6-methoxy-[1,3]thiazolo[4,5-b]pyridin-2-yl}-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000217_0001
In a reaction vial, 6-methoxy-N-[(pyridin-3-yl)methyl]-[1,3]thiazolo[4,5-b]pyridin-2-amine (145 mg, 0.53 mmol) and N,N-diisopropylethylamine (0.27 mL, 1.6 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (116 mg, 0.79 mmol) was added dropwise with stirring and the reaction was stirred for 16 hours. The product was confirmed present via UPLC analysis. 10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-{6-methoxy-[1,3]thiazolo[4,5-b]pyridin-2-yl}-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide (117 mg, 0.31 mmol, 58 %) was afforded as a yellow solid. 1H NMR (300 MHz, Chloroform-d) δ 8.59 (d, J = 1.9 Hz, 1H), 8.55 (dd, J = 4.8, 1.5 Hz, 1H), 8.34 (d, J = 2.9 Hz, 1H), 7.79 – 7.54 (m, 2H), 7.27 (ddd, J = 7.8, 4.9, 0.9 Hz, 1H), 5.69 (s, 2H), 3.93 (d, J = 1.0 Hz, 3H), 2.71 (tt, J = 11.1, 3.2 Hz, 1H), 1.97 – 1.55 (m, 7H), 1.24 (t, J = 10.5 Hz, 3H); LCMS (ESI) m/z: 383.1 [M+H]+. Example 175. Preparation of N-(6-bromo-1,3-benzoxazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000217_0002
Step 1: Preparation of 6-bromo-N-[(pyridin-3-yl)methyl]-1,3-benzoxazol-2-amine
Figure imgf000217_0003
In a round bottom flask equipped with a stir bar, a solution of pyridine-3-carbaldehyde (0.22 mL, 2.3 mmol) and 6-bromo-1,3-benzoxazol-2-amine (500 mg, 2.3 mmol) in dry toluene (12 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (265 mg, 7.0 mmol) in dry ethyl alcohol (12 mL). The reaction mixture was stirred at 70 °C for 1 hour and then cooled to room temperature. The reaction was filtered over a bed of Celite and washed with ethyl acetate (2x, 20 mL). The filtrate was concentrated then diluted with dichloromethane and re-filtered. The filtrate was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane. Product 6-bromo-N- [(pyridin-3-yl)methyl]-1,3-benzoxazol-2-amine (293 mg, 0.96 mmol, 41 %) was afforded as a white solid. 1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 8.81 – 8.64 (m, 1H), 8.60 (dd, J = 2.3, 0.9 Hz, 1H), 8.48 (dd, J = 4.8, 1.7 Hz, 1H), 7.78 (dt, J = 7.9, 2.0 Hz, 1H), 7.66 (d, J = 1.9 Hz, 1H), 7.37 (ddd, J = 7.8, 4.7, 0.9 Hz, 1H), 7.28 (dd, J = 8.3, 1.9 Hz, 1H), 7.19 (d, J = 8.3 Hz, 1H), 4.54 (d, J = 5.3 Hz, 1H), 3.34 (s, 2H); LCMS (ESI) m/z: 306.00 [M+2H]+. Step 2: Preparation of N-(6-bromo-1,3-benzoxazol-2-yl)-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide
Figure imgf000218_0001
In a reaction vial, 6-bromo-N-[(pyridin-3-yl)methyl]-1,3-benzoxazol-2-amine (293 mg, 0.96 mmol) was added. was dissolved in tetrahydrofuran (5 mL) and cooled to 0 °C. To the solution was added triethylamine (0.20 mL, 1.4 mmol) then cyclohexanecarbonyl chloride (0.19 mL, 1.4 mmol) was added, slowly. The reaction was stirred from 0 °C to room temperature for 5 hours. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane and washed with water (3x). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Crude product was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane through 24 g of silica gel. Product N-(6-bromo-1,3-benzoxazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide (52.7 mg, 0.13 mmol, 13 %) was afforded as a white solid. 1H NMR (300 MHz, Chloroform-d) δ 8.75 – 8.47 (m, 2H), 7.81 – 7.60 (m, 2H), 7.63 – 7.39 (m, 2H), 7.36 – 7.16 (m, 1H), 5.19 (s, 2H), 3.27 (tt, J = 11.4, 3.3 Hz, 1H), 2.12 – 1.40 (m, 7H), 1.29 (d, J = 9.8 Hz, 3H); LCMS (ESI) m/z: 416.1 [M+2H]+. Example 176. Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-({4H,5H,6H-pyrrolo[1,2- b]pyrazol-3-yl}methyl)cyclohexanecarboxamide
Figure imgf000218_0002
Step 1: Preparation of N-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)methyl)-6-methoxybenzo[d]thiazol-2- amine
Figure imgf000218_0003
To a solution of 6-methoxybenzo[d]thiazol-2-amine (172 mg, 0.95 mmol), 5,6-dihydro-4H- pyrrolo[1,2-b]pyrazole-3-carbaldehyde (100 mg, 0.73 mmol) in toluene (8 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (111 mg, 2.9 mmol) in methanol (10 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane. Product N-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3- yl)methyl)-6-methoxybenzo[d]thiazol-2-amine (100 mg, 0.33 mmol, 45 %) was afforded as a white solid. LCMS (ESI) m/z: 301.0 [M+H]+. Step 2: Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-({4H,5H,6H-pyrrolo[1,2-b]pyrazol-3- yl}methyl)cyclohexanecarboxamide
Figure imgf000219_0001
In a reaction vial, N-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)methyl)-6- methoxybenzo[d]thiazol-2-amine (50.0 mg, 0.17 mmol) and triethylamine (51.0 mg, 0.50 mmol) were dissolved in dichloromethane (5 mL). Cyclohexanecarbonyl chloride (48.0 mg, 0.33 mmol) was added slowly, and the reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. Water (5 mL) was added and the reaction was extracted with 2x 10 mL portions of dichloromethane. The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by prep-HPLC to give the desired product N-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol- 3-yl)methyl)-N-(6-methoxybenzo[d]thiazol-2-yl)cyclohexanecarboxamide (5.00 mg, 0.010 mmol, 7.2%) as white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 7.60 (d, J =9.2 Hz, 1H), 7.40 (s, 1H), 7.31 (s, 1H), 6.96-6.93 (m, 1H), 5.31 (s, 2H), 3.93 (t, J =7.2 Hz, 2H), 3.76 (s, 3H), 2.89 - 2.87 (m, 1H), 3.14 (t, J =7.2 Hz, 2H), 2.45 - 2.39 (m, 2H), 1.74 - 1.62 (m, 5H), 1.50 - 1.41 (m, 2H), 1.29 - 1.17 (m, 3H); LCMS (ESI) m/z: 411.0 [M+H]+. Example 177. Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-({5H,6H,7H-pyrrolo[1,2- a]imidazol-3-yl}methyl)cyclohexanecarboxamide
Figure imgf000219_0002
Step 1: Preparation of N-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)methyl)-6-methoxybenzo[d]thiazol-2- amine
Figure imgf000219_0003
To a solution of 6-methoxybenzo[d]thiazol-2-amine (120 mg, 0.67 mmol), 6,7-dihydro-5H- pyrrolo[1,2-a]imidazole-3-carbaldehyde (70.0 mg, 0.51 mmol) in toluene (8 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (78 mg, 2.1 mmol) in methanol (10 mL) was added. The reaction was stirred at room temperature for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane. Product N-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3- yl)methyl)-6-methoxybenzo[d]thiazol-2-amine (100 mg, 0.33 mmol, 52 %) was afforded as a white solid. LCMS (ESI) m/z: 301.0 [M+H]+. Step 2: Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-({5H,6H,7H-pyrrolo[1,2-a]imidazol-3- yl}methyl)cyclohexanecarboxamide
Figure imgf000220_0001
In a reaction vial, a mixture of cyclohexanecarboxylic acid (42.0 mg, 0.33 mmol), N,N- diisopropylethylamine (0.17 mL, 1.0 mmol) and N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (127 mg, 0.33 mmol) in tetrahydrofuran (10 mL) was stirred at room temperature for 30 minutes. Then, N-((6,7-dihydro-5H- pyrrolo[1,2-a]imidazol-3-yl)methyl)-6-methoxybenzo[d]thiazol-2-amine (100 mg, 0.33 mmol) was added and the reaction mixture was stirred at room temperature for 1.5 hours. The product was indicated present via UPLC analysis. The mixture was concentrated under reduced pressure. Crude product was purified by prep-HPLC to give the desired product N-(6-methoxy-1,3-benzothiazol-2-yl)-N-({5H,6H,7H- pyrrolo[1,2-a]imidazol-3-yl}methyl)cyclohexanecarboxamide (17.5 mg, 0.043 mmol, 13 %) as yellow solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 7.72 (d, J = 8.0 Hz, 1H), 7.43 (s, 1H), 7.07 (d, J = 8.0 Hz, 1H), 6.87 (s, 1H), 5.58 (s, 2H), 3.91 (t, J = 7.2 Hz, 2H), 3.89 (s, 3H), 3.00-2.98 (m, 1H), 2.80 (t, J = 7.2 Hz, 2H), 2.60 - 2.53 (m, 2H), 1.83 - 1.76 (m, 5H), 1.61 - 1.52 (m, 2H), 1.34 - 1.28 (m, 3H); LCMS (ESI) m/z: 411.0 [M+H]+. Example 178. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]oxane-4- carboxamide
Figure imgf000220_0002
Step 1: Preparation of 6-ethyl-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine
Figure imgf000221_0001
To a solution of 6-ethylbenzo[d]thiazol-2-amine (100 mg, 0.56 mmol), 1-trityl-1H-imidazole-4- carbaldehyde (247 mg, 0.73 mmol) in toluene (8 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (78 mg, 2.1 mmol) in methanol (10 mL) was added. The reaction was stirred at room temperature for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane. Product 6-ethyl-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (70.0 mg, 0.14 mmol, 25 %) was afforded as a white solid. LCMS (ESI) m/z: 501.0 [M+H]+. Step 2: Preparation of N-(6-ethylbenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4-yl)methyl)tetrahydro-2H- pyran-4-carboxamide
Figure imgf000221_0002
To a mixture of 6-ethyl-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (70.0 mg, 0.14 mmol) and triethylamine (51.0 mg, 0.50 mmol) in dichloromethane (5 mL) was added cyclohexanecarbonyl chloride (48.0 mg, 0.33 mmol). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The mixture was concentrated and purified by flash column chromatography through silica gel eluting with 0-5% methanol in dichloromethane to give the desired product N-(6-ethylbenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4-yl)methyl)tetrahydro-2H- pyran-4-carboxamide (21.5 mg, 0.035 mmol, 25 %) as white solid. LCMS (ESI) m/z: 614.0 [M+H]+. Step 3: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]oxane-4-carboxamide
Figure imgf000221_0003
To a solution of N-(6-ethylbenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4-yl)methyl)tetrahydro-2H- pyran-4-carboxamide (21.5 mg, 0.035 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (2 mL), and the reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The resulting mixture was concentrated and quenched with a solution of 7 M ammonia in methanol, until pH 8. The mixture was concentrated and purified directly by prep-HPLC (the crude samples were dissolved in methanol before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous ammonium bicarbonate). Product N-(6-ethyl-1,3- benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]oxane-4-carboxamide (6.00 mg, 0.016 mmol, 46 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 12.00 (s, 1H), 7.77 (s, 1H), 7.71 (d, J = 8.0 Hz, 1H), 7.58 (s, 1H), 7.28 (d, J = 8.0 Hz, 1H), 7.08 (s, 1H), 5.49 (s, 2H), 3.91 - 3.89 (m, 2H), 3.64 - 3.63 (m, 1H), 3.43 - 3.33 (m, 2H), 2.74 - 2.68 (m, 2H), 1.73 - 1.71 (m, 4H) 1.22 (t, J = 8.0 Hz, 3H); LCMS (ESI) m/z: 371.0 [M+H]+. Example 179. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-2-phenoxy-N-[(pyridin-3- yl)methyl]acetamide
Figure imgf000222_0001
Step 1: Preparation of 5-fluoro-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000222_0002
In a round bottom flask equipped with a stir bar, a solution of pyridine-3-carbaldehyde (1.38 mL, 15 mmol) and 5-fluoro-1,3-benzothiazol-2-amine (2.50 g, 15 mmol) in dry toluene (74 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen overnight. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (2.80 g, 74 mmol) in dry ethyl alcohol (74 mL). The reaction mixture was stirred at 70 °C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with ethyl acetate. The filtrate was concentrated then diluted with dichloromethane and filtered again. The filtrate was purified via flash column chromatography eluting with 0-100% hexanes/dichloromethane. Product 5-fluoro-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (1.26 g, 4.9 mmol, 33 %) was afforded as a white powder.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 8.95 – 8.33 (m, 2H), 7.91 – 7.59 (m, 2H), 7.29 (ddd, J = 47.6, 9.2, 3.7 Hz, 2H), 6.88 (td, J = 9.1, 2.6 Hz, 1H), 4.63 (d, J = 4.7 Hz, 2H), 3.36 (s, 1H); LCMS (ESI) m/z: 261.0 [M+H]+. Step 2: Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-2-phenoxy-N-[(pyridin-3-yl)methyl]acetamide
Figure imgf000222_0003
In a reaction vial, 5-fluoro-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (275 mg, 1.1 mmol) was added. was dissolved in tetrahydrofuran (5.30 mL) and cooled to 0 °C. To the solution was added triethylamine (0.29 mL, 2.12 mmol) then 2-phenoxyacetyl chloride (0.25 mL, 1.63 mmol) was added, slowly. The reaction was stirred from 0 °C to room temperature for 2 hours. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane and washed with water (3x). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Crude product was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane through 24 g of silica gel. Product N-(5-fluoro-1,3-benzothiazol-2-yl)-2-phenoxy-N-[(pyridin-3-yl)methyl]acetamide (61.0 mg, 0.16 mmol, 15 %) was afforded as a pale yellow solid.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 8.71 – 8.44 (m, 1H), 8.15 – 7.96 (m, 1H), 7.82 – 7.55 (m, 1H), 7.32 (dt, J = 33.5, 6.6 Hz, 2H), 7.02 – 6.79 (m, 2H), 5.62 (s, 1H), 5.19 (s, 1H), 3.34 (s, 7H); LCMS (ESI) m/z: 394.1 [M+H]+. Example 180. Preparation of N-[(1H-imidazol-4-yl)methyl]-N-(5-methoxy-1,3-benzothiazol-2-yl)-3- phenoxypropanamide
Figure imgf000223_0001
Step 1: Preparation of N-[(1H-imidazol-5-yl)methyl]-5-methoxy-1,3-benzothiazol-2-amine
Figure imgf000223_0002
In a reaction vial, a solution of 5-methoxy-1,3-benzothiazol-2-amine (200 mg, 1.1 mmol) and 1H- imidazole-5-carbaldehyde (105 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (95.7 mg, 2.5 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-20% methanol in ethyl acetate. Product N-[(1H-imidazol-5-yl)methyl]-5-methoxy-1,3-benzothiazol-2-amine (87.3 mg, 0.34 mmol, 31 %) was afforded as an orange solid.1H NMR (300 MHz, Methanol-d4) δ 7.69 (s, 1H), 7.43 (dd, J = 8.7, 1.6 Hz, 1H), 7.09 (s, 1H), 7.04 (t, J = 1.9 Hz, 1H), 6.70 (dt, J = 8.6, 2.0 Hz, 1H), 4.57 (s, 2H), 3.81 (d, J = 1.5 Hz, 3H); LCMS (ESI) m/z: 261.0 [M+H]+. Step 2: Preparation of N-[(1H-imidazol-4-yl)methyl]-N-(5-methoxy-1,3-benzothiazol-2-yl)-3- phenoxypropanamide
Figure imgf000224_0001
In a reaction vial, 3-phenoxypropanoic acid (111 mg, 0.67 mmol) was dissolved in thionyl chloride (0.24 mL, 3.4 mmol) and stir at room temperature for 2 hours. The reaction was concentrated under reduced pressure and re-dissolved in dimethylformamide (1 mL). N,N-diisopropylethylamine (0.13 mL, 0.74 mmol) and N-[(1H-imidazol-4-yl)methyl]-5-methoxy-1,3-benzothiazol-2-amine (87.3 mg, 0.34 mmol) were added at 0 °C with stirring. The reaction was warmed to room temperature and stirred for 2 hours. The product was indicated present via UPLC analysis. The reaction was diluted with 10 mL deionized water and 10 mL ethyl acetate. The organic layer was separated and washed with 2x 10 mL portions of deionized water, then concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-[(1H-imidazol-4-yl)methyl]-N-(5-methoxy-1,3-benzothiazol-2-yl)-3- phenoxypropanamide (34.7 mg, 0.085 mmol, 26 %) was afforded as an orange solid.1H NMR (300 MHz, Chloroform-d) δ 7.62 (d, J = 8.7 Hz, 1H), 7.55 (s, 1H), 7.33 (s, 1H), 7.22 (d, J = 8.2 Hz, 2H), 7.11 (s, 1H), 6.97 – 6.80 (m, 4H), 5.44 (s, 2H), 4.37 (t, J = 6.5 Hz, 2H), 3.85 (q, J = 1.7 Hz, 3H), 3.37 (t, J = 6.0 Hz, 2H); LCMS (ESI) m/z: 409.1 [M+H]+. Example 181. Preparation of N-(1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]-3- phenoxypropanamide
Figure imgf000224_0002
Step 1: Preparation of N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000224_0003
In a reaction vial, a solution of 1H-imidazole-5-carbaldehyde (127 mg, 1.3 mmol) and 1,3- benzothiazol-2-amine (200 mg, 1.3 mmol) in dry toluene (13 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (115 mg, 3.1 mmol) in dry ethyl alcohol (13 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-20% methanol in ethyl acetate. Product N-[(1H-imidazol-5- yl)methyl]-1,3-benzothiazol-2-amine (235 mg, 1.0 mmol, 77 %) was afforded as an orange solid.1H NMR (300 MHz, Methanol-d4) δ 7.66 (d, J = 1.6 Hz, 1H), 7.63 – 7.53 (m, 1H), 7.45 (d, J = 8.0 Hz, 1H), 7.34 – 7.20 (m, 1H), 7.15 – 6.99 (m, 2H), 4.58 (s, 2H); LCMS (ESI) m/z: 230.9 [M+H]+. Step 2: Preparation of N-(1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]-3-phenoxypropanamide
Figure imgf000225_0001
In a reaction vial, 3-phenoxypropanoic acid (338 mg, 2.0 mmol) was dissolved in thionyl chloride (0.74 mL, 10 mmol) and stir at room temperature for 2 hours. The reaction was concentrated under reduced pressure and re-dissolved in dimethylformamide (1.3 mL). N,N-diisopropylethylamine (0.39 mL, 2.2 mmol) and N-[(1H-imidazol-4-yl)methyl]-1,3-benzothiazol-2-amine (236 mg, 1.0 mmol) were added at 0 °C with stirring. The reaction was warmed to room temperature and stirred for 2 hours. The product was indicated present via UPLC analysis. The reaction was diluted with 10 mL deionized water and 10 mL ethyl acetate. The organic layer was separated and washed with 2x 10 mL portions of deionized water, then concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-(1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]-3-phenoxypropanamide (82.7 mg, 0.22 mmol, 21 %) was afforded as an orange solid.1H NMR (300 MHz, Chloroform-d) δ 7.89 (d, J = 8.1 Hz, 1H), 7.83 (dd, J = 8.0, 1.2 Hz, 1H), 7.60 (s, 1H), 7.47 (td, J = 8.2, 7.8, 1.3 Hz, 1H), 7.38 – 7.26 (m, 3H), 7.17 (s, 1H), 7.01 – 6.89 (m, 3H), 5.51 (s, 2H), 4.44 (t, J = 6.4 Hz, 2H), 3.45 (t, J = 6.4 Hz, 2H); LCMS (ESI) m/z: 379.1 [M+H]+. Example 182. Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000225_0002
Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]cyclohexanecarboxamide
Figure imgf000225_0003
A mixture of N-((1H-imidazol-4-yl)methyl)-6-chlorobenzo[d]thiazol-2-amine (156 mg, 0.6 mmol), cyclohexanecarboxylic acid (92.0 mg, 0.72 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (456 mg, 1.2 mmol), N,N- diisopropylethylamine (180 mg, 1.8 mmol) in dichloromethane (10 mL) was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. The mixture was quenched with ice water, then extracted with ethyl acetate (2x 15 mL). The organic layers were pooled, washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (the crude samples were dissolved in methanol before purification. Boston C18 21*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous ammonium bicarbonate) to give the desired product N-((1H-imidazol-4-yl)methyl)-N-(6-chlorobenzo[d]thiazol-2- yl)cyclohexanecarboxamide (15.5 mg, 0.040 mmol, 6.9 %) as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 11.99 (s, 1H), 8.10 (d, J = 2.1 Hz, 1H), 7.80 (d, J = 8.7 Hz, 1H), 7.58 (s, 1H), 7.45 (dd, J = 8.6, 2.1 Hz, 1H), 7.07 (s, 1H), 5.45 (s, 2H), 1.85 – 1.62 (m, 5H), 1.50 – 1.21 (m, 6H); LCMS (ESI) m/z: 375.1 [M+H]+. Example 183. Preparation of methyl 4-{[(1H-imidazol-4-yl)methyl][6-(trifluoromethyl)-1,3- benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000226_0001
Step 1: Preparation of 6-(trifluoromethyl)-N-{[1-(triphenylmethyl)-1H-imidazol-4-yl]methyl}-1,3- benzothiazol-2-amine
Figure imgf000226_0002
In a reaction vial equipped with a magnetic stir bar, 1-(triphenylmethyl)-1H-imidazole-4- carbaldehyde (697 mg, 2.1 mmol) and 6-(trifluoromethyl)-1,3-benzothiazol-2-amine (450 mg, 2.1 mmol) were dissolved in dioxane (10 mL). To the reaction was added titanium(IV) isopropoxide (0.61 mL, 2.1 mmol) and the reaction was stirred at 60 °C under nitrogen for 1 hour. The reaction was cooled to room temperature and to the reaction was added sodium borohydride (155 mg, 4.1 mmol). The reaction mixture was stirred at room temperature for 4 hours. The product was indicated present via UPLC analysis. The reaction was quenched with methanol and stirred until the bubbling ceased, then concentrated under reduced pressure. Crude product was purified via flash column chromatography eluting with 0-100% ethyl acetate in hexanes. Product 6-(trifluoromethyl)-N-{[1-(triphenylmethyl)-1H-imidazol-4-yl]methyl}-1,3- benzothiazol-2-amine (453 mg, 0.84 mmol, 41 %) was afforded as a white solid.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 8.64 (t, J = 5.2 Hz, 1H), 8.12 (d, J = 1.6 Hz, 1H), 7.59 – 7.27 (m, 12H), 7.19 – 6.99 (m, 6H), 6.90 (d, J = 1.4 Hz, 1H), 4.45 (d, J = 5.2 Hz, 2H). Step 2: Preparation of methyl 4-{[6-(trifluoromethyl)-1,3-benzothiazol-2-yl]({[1-(triphenylmethyl)-1H- imidazol-4-yl]methyl})carbamoyl}piperidine-1-carboxylate
Figure imgf000227_0001
In a reaction vial equipped with a stir bar, 6-(trifluoromethyl)-N-{[1-(triphenylmethyl)-1H-imidazol- 4-yl]methyl}-1,3-benzothiazol-2-amine (454 mg, 0.84 mmol) was dissolved in dichloromethane (4 mL). To the reaction solution was added 1-(methoxycarbonyl)piperidine-4-carboxylic acid (312 mg, 1.7 mmol) and 50% propylphosphonic anhydride in ethyl acetate (1.6 mL, 2.1 mmol). The reaction mixture was cooled to 0 °C and triethylamine (0.58 mL, 4.2 mmol) was added. The reaction was stirred from 0 °C to room temperature overnight. The product was indicated present via UPLC analysis. The reaction solution was diluted with dichloromethane and washed with water (3 x 15 mL). The organic layers were pooled, dried over anhydrous sodium sulfate, filtered and concentrated. Crude product was purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane. Product methyl 4-{[6- (trifluoromethyl)-1,3-benzothiazol-2-yl]({[1-(triphenylmethyl)-1H-imidazol-4- yl]methyl})carbamoyl}piperidine-1-carboxylate (530 mg, 0.75 mmol, 89 %) was afforded as a white solid. 1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 8.48 (d, J = 1.8 Hz, 1H), 7.92 – 7.64 (m, 2H), 7.51 – 7.17 (m, 11H), 7.15 – 6.86 (m, 7H), 5.50 (s, 2H), 4.16 – 3.71 (m, 2H), 3.33 (s, 3H), 3.04 – 2.63 (m, 4H), 1.78 (d, J = 13.4 Hz, 3H), 1.67 – 1.29 (m, 4H). Step 3: Preparation of methyl 4-{[(1H-imidazol-4-yl)methyl][6-(trifluoromethyl)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000227_0002
In a reaction vial equipped with a stir bar, methyl 4-{[6-(trifluoromethyl)-1,3-benzothiazol-2-yl]({[1- (triphenylmethyl)-1H-imidazol-4-yl]methyl})carbamoyl}piperidine-1-carboxylate (531 mg, 0.75 mmol) was dissolved in dichloromethane (4 mL). To the reaction was added trifluoroacetic acid (0.46 mL, 6.0 mmol) and the reaction was stirred at room temperature overnight. The product was indicated present via UPLC analysis. The reaction was concentrated under reduced pressure then diluted with dichloromethane and quenched with saturated sodium bicarbonate until bubbling ceased. The reaction was diluted with 10 mL deionized water and washed with dichloromethane (2x 10 mL). The organic layers were combined and dried over anhydrous sodium sulfate, filtered, then concentrated. Crude product was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane. Product methyl 4-{[(1H- imidazol-4-yl)methyl][6-(trifluoromethyl)-1,3-benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate (159 mg, 0.34 mmol, 46 %) was afforded as a pale yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.11 (dt, J = 1.7, 0.8 Hz, 1H), 8.03 – 7.88 (m, 1H), 7.78 – 7.57 (m, 2H), 7.16 (s, 1H), 5.56 (s, 2H), 4.44 – 4.06 (m, 3H), 3.73 (s, 3H), 3.54 (q, J = 7.5 Hz, 1H), 2.98 (s, 3H), 2.08 (d, J = 12.7 Hz, 1H), 1.98 – 1.67 (m, 4H); LCMS (ESI) m/z: 468.2 [M+H]+. Example 184. Preparation of 1-acetyl-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]piperidine-4-carboxamide
Figure imgf000228_0001
Step 1: Preparation of 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000228_0002
In a round bottom flask equipped with a stir bar, a solution of pyridine-3-carbaldehyde (0.63 mL, 6.7 mmol) and 6-ethyl-1,3-benzothiazol-2-amine (1.20 g, 6.7 mmol) in dry toluene (34 mL) was stirred at 105°C with activated 4 Å molecular sieves under nitrogen overnight. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (1.27 g, 34 mmol) in dry ethyl alcohol (34 mL). The reaction mixture was stirred at 70 °C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with ethyl acetate. The filtrate was concentrated then diluted with dichloromethane and filtered again. The filtrate was purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane through a 40 g column of silica gel. Product 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3- benzothiazol-2-amine (0.915 g, 3.4 mmol, 51 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.72 – 8.50 (m, 2H), 7.76 (dddd, J = 7.9, 2.4, 1.7, 0.8 Hz, 1H), 7.50 – 7.37 (m, 2H), 7.35 – 7.22 (m, 1H), 7.14 (dt, J = 8.3, 1.2 Hz, 1H), 6.24 (s, 1H), 4.68 (s, 2H), 2.70 (q, J = 7.6 Hz, 2H), 1.26 (td, J = 7.6, 0.7 Hz, 3H); LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of 1-acetyl-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperidine-4- carboxamide
Figure imgf000228_0003
In a reaction vial equipped with a stir bar, 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2- amine (258 mg, 0.96 mmol) was dissolved in dichloromethane (5 mL). To the reaction solution was added 1-acetylpiperidine-4-carboxylic acid (326 mg, 1.9 mmol) and 50% propylphosphonic anhydride in ethyl acetate (1.81 mL, 2.4 mmol). The reaction mixture was cooled to 0 °C and triethylamine (0.67 mL, 4.8 mmol) was added. The reaction was stirred from 0 °C to room temperature for 3 days. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane and washed with water (3 x 15 mL). The organic layers were pooled, dried over anhydrous sodium sulfate, filtered and concentrated. Crude product was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane. Product 1-acetyl-N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperidine- 4-carboxamide (135 mg, 0.32 mmol, 33 %) was afforded as yellow solid.1H NMR (300 MHz, Chloroform- d) δ 8.70 – 8.50 (m, 2H), 7.81 – 7.51 (m, 3H), 7.40 – 7.18 (m, 2H), 4.62 (d, J = 13.7 Hz, 1H), 3.87 (d, J = 14.0 Hz, 1H), 3.03 (t, J = 13.9 Hz, 2H), 2.78 (q, J = 7.6 Hz, 2H), 2.60 (dd, J = 14.4, 7.5 Hz, 1H), 2.10 (s, 9H), 1.30 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 423.2 [M+H]+. Example 185. Preparation of N-(6-cyano-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000229_0001
Step 1: Preparation of 2-(((1-trityl-1H-imidazol-4-yl)methyl)amino)benzo[d]thiazole-6-carbonitrile
Figure imgf000229_0002
To a solution of 6-cyanobenzo[d]thiazol-2-amine (100 mg, 0.56 mmol), 1-trityl-1H-imidazole-4- carbaldehyde (247 mg, 0.73 mmol) in toluene (16 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (78.0 mg, 2.1 mmol) in methanol (30 mL) was added. The reaction was stirred at room temperature for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane. Product 2-(((1-trityl-1H-imidazol-4-yl)methyl)amino)benzo[d]thiazole-6-carbonitrile (70.0 mg, 0.14 mmol, 25 %) was afforded as a yellow solid. LCMS (ESI) m/z: 498.3 [M+H]+. Step 2: Preparation of N-(6-cyanobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide
Figure imgf000230_0001
In a reaction vial, a mixture cyclohexanecarbonyl chloride (39.0 mg, 0.30 mmol), N,N- diisopropylethylamine (0.10 mL, 0.48 mmol) and N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (115 mg, 0.30 mmol) in tetrahydrofuran (10 mL) was stirred at room temperature for 30 minutes. Then 2-(((1-trityl-1H-imidazol-4- yl)methyl)amino)benzo[d]thiazole-6-carbonitrile (60.0 mg, 0.12 mmol) was added to the reaction vial. The reaction was stirred at 70 °C for 2 hours. The product was indicated present via UPLC analysis. The reaction mixture was concentrated under reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane. Product N-(6- cyanobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4-yl)methyl)cyclohexanecarboxamide (60.0 mg, 0.099 mmol, 82 %) was afforded as a white solid. LCMS (ESI) m/z: 630.3 [M+Na]+. Step 3: Preparation of N-(6-cyano-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000230_0002
To a solution of 2-(((1-trityl-1H-imidazol-4-yl)methyl)amino)benzo[d]thiazole-6-carbonitrile (60 mg, 0.099 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1.5 mL). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The mixture was concentrated under reduced pressure and purified by prep-HPLC to give N-(6-cyano-1,3-benzothiazol-2- yl)-N-[(1H-imidazol-4-yl)methyl]cyclohexanecarboxamide (20.0 mg, 0.055 mmol, 55 %) as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 12.00 (s, 1H), 8.56 (s, 1H), 7.94 (d, J = 8.0 Hz, 1H), 7.83 (d, J = 8.0 Hz, 1H), 7.58 (s, 1H), 7.11 (s, 1H), 5.49 (s, 2H), 1.84 - 1.66 (m, 5H), 1.46 - 1.43 (m, 2H), 1.33 - 1.24 (m, 4H); LCMS (ESI) m/z: 366.0 [M+H]+. Example 186. Preparation of N-(6-hydroxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000230_0003
Step 1: Preparation of 2-((pyridin-3-ylmethyl)amino)benzo[d]thiazol-6-ol
Figure imgf000231_0001
To a solution of 2-aminobenzo[d]thiazol-6-ol (300 mg, 1.8 mmol), nicotinaldehyde (193 mg, 1.8 mmol) in toluene (40 mL) was added dried 4 Å molecular sieves. The mixture was heated to 120 °C and stirred for 16 hours. Then a solution of sodium borohydride (342 mg, 9.0 mmol) in methanol (30 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-10% methanol in dichloromethane. Product 2- ((pyridin-3-ylmethyl)amino)benzo[d]thiazol-6-ol (150 mg, 0.58 mmol, 32 %) was afforded as a white solid. Step 2: Preparation of N-(6-hydroxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000231_0002
In a reaction vial, a mixture of 2-((pyridin-3-ylmethyl)amino)benzo[d]thiazol-6-ol (100 mg, 0.39 mmol), cyclohexanecarboxylic acid (50.0 mg, 0.39 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5- b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (224 mg, 0.59 mmol), N,N-diisopropylethylamine (101 mg, 0.78 mmol) in tetrahydrofuran (10 mL) was stirred at 70 °C for 16 hours. The product was indicated present via UPLC analysis. The mixture was quenched with ice water, then extracted with ethyl acetate (2x 15 mL). The organic layers were pooled, washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (the crude samples were dissolved in methanol before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous ammonium bicarbonate) to give the desired product N-(6-hydroxybenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)cyclohexanecarboxamide (9.50 mg, 0.026 mmol, 6.6 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 9.70 (s, 1H), 8.54 (d, J = 1.6 Hz, 1H), 8.48 (dd, J = 1.6, 3.2 Hz, 1H), 7.62 (d, J = 2.0 Hz, 1H), 7.55 (d, J = 8.4 Hz, 1H), 7.38 (dd, J = 0.8, 4.0 Hz, 1H), 7.27 (d, J = 2.4 Hz, 1H), 6.86 (dd, J = 2.4, 6.4 Hz, 1H), 5.62 (s, 2H), 2.85-2.79 (m, 1H), 1.65 (d, J = 11.6 Hz, 5H), 1.46-1.18 (m, 5H); LCMS (ESI) m/z: 368.0 [M+H]+. Example 187. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]piperidine- 4-carboxamide formate
Figure imgf000231_0003
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]piperidine-4-carboxamide formate
Figure imgf000232_0001
In a reaction vial, tert-butyl 4-((6-ethylbenzo[d]thiazol-2-yl)(pyrimidin-5- ylmethyl)carbamoyl)piperidine-1-carboxylate (60.0 mg, 0.12 mmol) was dissolved in dichloromethane (3 mL). A 4 M solution of hydrochloric acid in dioxane (2 mL) was added to the reaction vial slowly, and the reaction was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The mixture was concentrated under reduced pressure and purified by prep-HPLC (Boston pHlex ODS 10um 21.2×250mm120A. The mobile phase was acetonitrile/0.1% formic acid) to give N-(6- ethylbenzo[d]thiazol-2-yl)-N-(pyrimidin-5-ylmethyl)piperidine-4-carboxamide formate (22.0 mg, 0.052 mmol, 43 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 9.11 (s, 1H), 8.79 (s, 2H), 7.82 (s, 1H), 7.65 (d, J = 8.3 Hz, 1H), 7.28 (d, J = 7.1 Hz, 1H), 5.63 (s, 2H), 3.21 - 3.07 (m, 3H), 2.74 – 2.67 (m, 4H), 1.80 – 1.67 (m, 4H), 1.22 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 382.0 [M+H]+. Example 188. Preparation of methyl 4-{[(pyridin-3-yl)methyl][6-(trifluoromethyl)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000232_0002
Step 1: Preparation of N-[(pyridin-3-yl)methyl]-6-(trifluoromethyl)-1,3-benzothiazol-2-amine
Figure imgf000232_0003
In a reaction vial, a solution of pyridine-3-carbaldehyde (0.12 mL, 1.3 mmol) and 6- (trifluoromethyl)-1,3-benzothiazol-2-amine (300 mg, 1.4 mmol) in dry toluene (7 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (119 mg, 3.2 mmol) in dry methanol (7 mL). The reaction mixture was stirred at 70 °C for 1 hour and then cooled to room temperature. The reaction was filtered over a bed of Celite and washed with methanol (2x 20 mL). The filtrate was concentrated then diluted with dichloromethane and re-filtered. Crude product was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane. Product N-[(pyridin-3-yl)methyl]-6- (trifluoromethyl)-1,3-benzothiazol-2-amine (289 mg, 0.94 mmol, 68 %) was afforded as a white solid.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 9.00 – 8.85 (m, 1H), 8.69 – 8.39 (m, 2H), 8.15 (s, 1H), 7.79 (dt, J = 7.8, 1.9 Hz, 1H), 7.53 (t, J = 1.5 Hz, 2H), 7.47 – 7.32 (m, 1H), 4.66 (d, J = 5.5 Hz, 2H), 3.34 (d, J = 1.7 Hz, 1H); LCMS (ESI) m/z: 310.0 [M+H]+. Step 2: Preparation of methyl 4-{[(pyridin-3-yl)methyl][6-(trifluoromethyl)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000233_0001
In a reaction vial equipped with a stir bar, N-[(pyridin-3-yl)methyl]-6-(trifluoromethyl)-1,3- benzothiazol-2-amine (290 mg, 0.93 mmol) was dissolved in dichloromethane (5 mL). To the reaction solution was added 1-(methoxycarbonyl)piperidine-4-carboxylic acid (348 mg, 1.9 mmol) and 50% propylphosphonic anhydride in ethyl acetate (1.76 mL, 2.3 mmol). The reaction mixture was cooled to 0 °C and triethylamine (0.65 mL, 4.7 mmol) was added. The reaction was stirred from 0 °C to room temperature over 3 days. The product was indicated present over UPLC analysis. The reaction solution was diluted with dichloromethane and washed with water (3x 15 mL). The organic layers were pooled, dried over anhydrous sodium sulfate, filtered and concentrated. Purified via flash column chromatography eluting with 0-10% methanol in dichloromethane. Product methyl 4-{[(pyridin-3-yl)methyl][6- (trifluoromethyl)-1,3-benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate (121 mg, 0.25 mmol, 27 %) was afforded as a pale yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.74 – 8.52 (m, 1H), 8.20 – 8.05 (m, 1H), 7.86 (d, J = 8.5 Hz, 1H), 7.75 – 7.55 (m, 1H), 7.42 – 7.22 (m, 1H), 5.69 (s, 1H), 4.01 (d, J = 37.5 Hz, 3H), 3.09 – 2.70 (m, 4H), 2.53 (tt, J = 10.8, 3.9 Hz, 1H), 2.11 – 1.47 (m, 6H); LCMS (ESI) m/z: 479.1 [M+H]+. Example 189. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(5-methylpyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000233_0002
Step 1: Preparation of 6-ethyl-N-[(5-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000233_0003
In a reaction vial, a solution of 5-methylpyridine-3-carbaldehyde (135 mg, 1.1 mmol) and 6-ethyl- 1,3-benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry methanol (11 mL). The resultant reaction mixture was refluxed at 100 °C for 1 hour and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-ethyl-N-[(5-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (260 mg, 0.92 mmol, 82 %) was afforded as a cream solid.1H NMR (300 MHz, Methanol-d4) δ 8.38 (s, 1H), 8.29 (s, 1H), 7.71 (s, 1H), 7.43 (s, 1H), 7.34 (d, J = 8.2 Hz, 1H), 7.11 (d, J = 8.2 Hz, 1H), 4.64 (s, 2H), 2.66 (q, J = 7.6 Hz, 2H), 2.36 (s, 3H), 1.23 (td, J = 7.6, 1.2 Hz, 3H); LCMS (ESI) m/z: 284.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(5-methylpyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000234_0001
In a reaction vial, 6-ethyl-N-[(5-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (261 mg, 0.92 mmol) and N,N-diisopropylethylamine (0.48 mL, 2.8 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (200 mg, 1.4 mmol) was added dropwise with stirring and the reaction was stirred for 16 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(5-methylpyridin-3- yl)methyl]cyclohexanecarboxamide (329 mg, 0.84 mmol, 91 %) was afforded as a yellow solid. 1H NMR (300 MHz, Chloroform-d) δ 8.39 (d, J = 12.5 Hz, 2H), 7.69 (d, J = 8.3 Hz, 1H), 7.64 (d, J = 1.7 Hz, 1H), 7.38 (s, 1H), 7.26 (dd, J = 8.3, 1.8 Hz, 1H), 5.58 (s, 2H), 2.77 (q, J = 7.6 Hz, 2H), 2.72 – 2.59 (m, 1H), 2.29 (s, 3H), 1.83 – 1.58 (m, 7H), 1.29 (t, J = 7.6 Hz, 3H), 1.26 – 1.11 (m, 3H); LCMS (ESI) m/z: 394.2 [M+H]+. Example 190. Preparation of N-(6-fluoro-1H-1,3-benzodiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000234_0002
Step 1: Preparation of 6-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)-1H-benzo[d]imidazol-2-amine
Figure imgf000235_0001
To a solution of 6-fluoro-1H-benzo[d]imidazol-2-amine (0.500 g, 3.3 mmol), 1-trityl-1H-imidazole- 4-carbaldehyde (1.10 g, 3.3 mmol) in toluene (40 mL) was added dried 4 Å molecular sieves. The mixture was heated to 120 °C and stirred for 16 hours. Then a solution of sodium borohydride (0.342 g, 9.0 mmol) in methanol (30 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-25% ethyl acetate in dichloromethane. Product 6-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)-1H-benzo[d]imidazol-2-amine (0.350 g, 0.74 mmol, 22 %) as a white solid. LCMS (ESI) m/z: 474.0 [M+H]+. Step 2: Preparation of N-(6-fluoro-1H-benzo[d]imidazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)cyclohexanecarboxamide
Figure imgf000235_0002
In a reaction vial, a mixture of 6-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)-1H-benzo[d]imidazol- 2-amine (350 mg, 0.74 mmol), cyclohexanecarboxylic acid (95.0 mg, 0.74 mmol), N-[(dimethylamino)-1H- 1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (418 mg, 1.1 mmol), N,N-diisopropylethylamine (194 mg, 1.5 mmol) in tetrahydrofuran (15 mL) was stirred at 70 °C for 16 hours. The product was indicated present via UPLC analysis. The mixture was quenched with ice water, then extracted with ethyl acetate (2x 15 mL). The organic layers were pooled, washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-25% ethyl acetate in petroleum ether. Product N-(6-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-5- yl)methyl)cyclohexanecarboxamide (400 mg, 0.68 mmol, 93 %) was afforded as a light yellow solid. LCMS (ESI) m/z: 584.0 [M+H]+. Step 3: Preparation of N-(6-fluoro-1H-1,3-benzodiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]cyclohexanecarboxamide
Figure imgf000236_0001
To a solution of N-(6-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-5- yl)methyl)cyclohexanecarboxamide (400 mg, 0.68 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (2 mL). The reaction was stirred at room temperature for 3 hours. The product was indicated present via UPLC analysis. The mixture was concentrated and quenched with a 7 M solution of ammonia in methanol until pH 8. The mixture was concentrated and purified by prep-HPLC to give product N-((1H-imidazol-5-yl)methyl)-N-(6-fluorobenzo[d]thiazol-2-yl)cyclohexanecarboxamide (35.3 mg, 0.10 mmol, 15 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 12.46 (s, 1H), 11.96 (s, 1H), 7.57 (s, 1H), 7.49 (d, J = 7.3 Hz, 1H), 7.30 (s, 1H), 6.97 (s, 2H), 5.17 (s, 2H), 1.89 – 0.90 (m, 11H); LCMS (ESI) m/z: 342.0 [M+H]+. Example 191. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-2-(propan-2-yloxy)-N-[(pyridin-3- yl)methyl]acetamide
Figure imgf000236_0002
Step 1: Preparation of 5-fluoro-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000236_0003
In a round bottom flask equipped with a stir bar, a solution of pyridine-3-carbaldehyde (1.38 mL, 15 mmol) and 5-fluoro-1,3-benzothiazol-2-amine (2.50 g, 15 mmol) in dry toluene (74 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen overnight. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (2.79 g, 74 mmol) in dry ethyl alcohol (74 mL). The reaction mixture was stirred at 70 °C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with ethyl acetate. The filtrate was concentrated then diluted with dichloromethane and filtered again. The filtrate was purified via flash column chromatography eluting with 0-100% hexanes/dichloromethane. Product 5-fluoro-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (1.26 g, 4.9 mmol, 33 %) was afforded as a white powder.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 8.95 – 8.33 (m, 2H), 7.91 – 7.59 (m, 2H), 7.29 (ddd, J = 47.6, 9.2, 3.7 Hz, 2H), 6.88 (td, J = 9.1, 2.6 Hz, 1H), 4.63 (d, J = 4.7 Hz, 2H), 3.36 (s, 1H); LCMS (ESI) m/z: 261.0 [M+H]+. Step 2: Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-2-(propan-2-yloxy)-N-[(pyridin-3- yl)methyl]acetamide
Figure imgf000237_0001
In a reaction vial equipped with a stir bar, 5-fluoro-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2- amine (250 mg, 0.96 mmol) was dissolved in dichloromethane (5 mL). To the reaction solution was added 2-(propan-2-yloxy)acetic acid (226 mg, 1.9 mmol) and 50% propylphosphonic anhydride in ethyl acetate (1.82 mL, 2.4 mmol). The reaction mixture was cooled to 0 °C and triethylamine (0.67 mL, 4.8 mmol) was added. The reaction was stirred from 0 °C to room temperature over 3 days. The product was indicated present via UPLC analysis. The reaction solution was diluted with dichloromethane and washed with water (3x 15 mL). The organic layers were pooled, dried over anhydrous sodium sulfate, filtered and concentrated. Purified via flash column chromatography eluting with 0-100% dichloromethane in ethyl acetate. Product N-(5-fluoro-1,3-benzothiazol-2-yl)-2-(propan-2-yloxy)-N-[(pyridin-3-yl)methyl]acetamide (250 mg, 0.70 mmol, 72 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.71 – 8.49 (m, 2H), 7.75 (dd, J = 8.7, 5.2 Hz, 1H), 7.60 (dddd, J = 7.9, 2.4, 1.7, 0.9 Hz, 1H), 7.47 (dd, J = 9.7, 2.5 Hz, 1H), 7.35 – 7.20 (m, 1H), 7.10 (td, J = 8.8, 2.4 Hz, 1H), 5.64 (s, 2H), 4.38 (s, 2H), 3.73 (p, J = 6.1 Hz, 1H), 1.19 (d, J = 6.0 Hz, 6H); LCMS (ESI) m/z: 360.2 [M+H]+. Example 192. Preparation of tert-butyl 4-{[(pyridin-3-yl)methyl][6-(trifluoromethoxy)-1,3- benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000237_0002
Step 1: Preparation of N-(pyridin-3-ylmethyl)-6-(trifluoromethoxy)benzo[d]thiazol-2-amine
Figure imgf000237_0003
To a solution of 6-(trifluoromethoxy)benzo[d]thiazol-2-amine (400 mg, 1.7 mmol) and nicotinaldehyde (183 mg, 1.7 mmol) in toluene (15 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (323 mg, 8.6 mmol) in ethanol (15 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane. Product N-(pyridin-3-ylmethyl)-6-(trifluoromethoxy)benzo[d]thiazol-2-amine (270 mg, 0.83 mmol, 49 %) was afforded as a white solid. LCMS (ESI) m/z: 326.0 [M+H]+. Step 2: Preparation of tert-butyl 4-{[(pyridin-3-yl)methyl][6-(trifluoromethoxy)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000238_0001
To a solution of N-(pyridin-3-ylmethyl)-6-(trifluoromethoxy)benzo[d]thiazol-2-amine (250 mg, 0.77 mmol) in N,N-dimethylformamide (15 mL) were added 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (263 mg, 1.2 mmol) and N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (437 mg, 1.2 mmol) and triethylamine (233 mg, 2.3 mmol). The reaction mixture was stirred at 45°C for 17 hours. The product was indicated present via UPLC analysis. The mixture was filtered over Celite and washed with 2x 15 mL portions of ethyl acetate. The filtrate was collected and the solvent was removed under the reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product tert-butyl 4-((pyridin-3-ylmethyl)(6-(trifluoromethoxy)benzo[d]thiazol-2- yl)carbamoyl)piperidine-1-carboxylate (59.9 mg, 0.11 mmol, 15 %) as a white solid.1H NMR (500 MHz, Dimethylsulfoxide-d6) δ 8.61 (s, 1H), 8.51 (d, J = 3.8 Hz, 1H), 8.15 (t, J = 4.1 Hz, 1H), 7.83 (d, J = 8.8 Hz, 1H), 7.68 (d, J = 8.0 Hz, 1H), 7.48 – 7.17 (m, 2H), 5.72 (s, 2H), 3.92 (s, 2H), 3.17 (dd, J = 15.1, 7.3 Hz, 1H), 2.70 (d, J = 65.8 Hz, 2H), 1.67 (d, J = 12.4 Hz, 2H), 1.50 (qd, J = 12.8, 4.2 Hz, 2H), 1.40 (s, 9H); LCMS (ESI) m/z: 537.0 [M+H]+. Example 193. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]morpholine-4-carboxamide
Figure imgf000238_0002
Step 1: Preparation of 6-ethyl-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine
Figure imgf000238_0003
To a solution of 6-ethylbenzo[d]thiazol-2-amine (100 mg, 0.56 mmol), 1-trityl-1H-imidazole-4- carbaldehyde (247 mg, 0.73 mmol) in toluene (8 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (78 mg, 2.1 mmol) in methanol (10 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane. Product 6-ethyl-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (200 mg, 0.43 mmol, 77 %) was afforded as a white solid. LCMS (ESI) m/z: 501.0 [M+H]+. Step 2: Preparation of N-(6-ethylbenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4-yl)methyl)morpholine-4- carboxamide
Figure imgf000239_0001
In a reaction vial, a mixture of morpholine (42.0 mg, 0.50 mmol), triethylamine (0.2 mL, 1.2 mmol) and triphosgene (119 mg, 0.40 mmol) in tetrahydrofuran (10 mL) was stirred at room temperature for 30 minutes. Then, Product 6-ethyl-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2-amine (200 mg, 0.43 mmol) was added and the reaction was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The mixture was concentrated under reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane. Product N-(6-ethylbenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)morpholine-4-carboxamide (80.0 mg, 0.13 mmol, 30%) was afforded as a yellow solid. LCMS (ESI) m/z: 614.5 [M+H]+. Step 3: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]morpholine-4- carboxamide
Figure imgf000239_0002
To a solution of N-(6-ethylbenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4-yl)methyl)morpholine-4- carboxamide (80.0 mg, 0.13 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1.5 mL). The reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The mixture was concentrated under reduced pressure and purified by prep-HPLC to give N-(6- ethyl-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]morpholine-4-carboxamide (12.0 mg, 0.032 mmol, 25 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 7.65 (s, 1H), 7.62-7.60 (m, 2H), 7.27-7.25 (m, 1H), 7.10 (s, 1H), 5.06 (s, 2H), 3.62 (t, J = 4.8 Hz, 4H), 3.48 (t, J = 4.8 Hz, 4H), 2.75-2.73 (m, 2H), 1.28 (t, J = 8.0 Hz, 3H); LCMS (ESI) m/z: 372.3 [M+H]+. Example 194. Preparation of N-[(pyridin-3-yl)methyl]-N-[6-(trifluoromethyl)-1,3-benzothiazol-2- yl]cyclohexanecarboxamide
Figure imgf000240_0001
Step 1: Preparation of N-(pyridin-3-ylmethyl)-6-(trifluoromethyl)benzo[d]thiazol-2-amine
Figure imgf000240_0002
To a solution of 6-(trifluoromethyl)benzo[d]thiazol-2-amine (200 mg, 0.90 mmol), nicotinaldehyde (108 mg, 1.0 mmol) in toluene (30 mL) was added dried 4 Å molecular sieves. The mixture was heated to 120 °C and stirred for 16 hours. Then a solution of sodium borohydride (175 mg, 4.6 mmol) in methanol (20 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-30% ethyl acetate in dichloromethane. Product N-(pyridin-3-ylmethyl)-6-(trifluoromethyl)benzo[d]thiazol-2-amine (120 mg, 0.39 mmol, 43 %) was afforded as a white solid. LCMS (ESI) m/z: 310.0 [M+H]+. Step 2: Preparation of N-[(pyridin-3-yl)methyl]-N-[6-(trifluoromethyl)-1,3-benzothiazol-2- yl]cyclohexanecarboxamide
Figure imgf000240_0003
In a reaction vial, a mixture of ethyl N-(pyridin-3-ylmethyl)-6-(trifluoromethyl)benzo[d]thiazol-2- amine (100 mg, 0.32 mmol), cyclohexanecarboxylic acid (50.0 mg, 0.39 mmol), N,N- diisopropylethylamine (84.0 mg, 0.65 mmol) and 1 N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (185 mg, 0.49 mmol) in tetrahydrofuran (5 mL) was stirred at 70 °C for 16 hours. The product was indicated present via UPLC analysis. The mixture was poured into water (20 mL) and extracted with 3x 15 mL washes of ethyl acetate. The organic phases were pooled, washed with brine (10 mL), dried over sodium sulfate, and concentrated under reduced pressure. Crude product was purified by prep-HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to afford N-(pyridin-3-ylmethyl)-N-(6-(trifluoromethyl)benzo[d]thiazol-2- yl)cyclohexanecarboxamide (13.3 mg, 0.032 mmol, 9.9 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.59 (d, J = 1.8 Hz, 1H), 8.53 (s, 1H), 8.50 (dd, J = 4.8, 1.5 Hz, 1H), 7.92 (d, J = 8.5 Hz, 1H), 7.73 (dd, J = 8.5, 1.7 Hz, 1H), 7.65 (d, J = 8.2 Hz, 1H), 7.37 (dd, J = 7.9, 4.8 Hz, 1H), 5.73 (s, 2H), 2.91 (t, J = 11.3 Hz, 1H), 1.67 (d, J = 9.4 Hz, 5H), 1.43 (d, J = 11.2 Hz, 2H), 1.24 (d, J = 13.2 Hz, 3H); LCMS (ESI) m/z: 420.1 [M+H]+. Example 195. Preparation of N-(6-ethoxy-1,3-benzothiazol-2-yl)-1-fluoro-N-[(pyridin-3- yl)methyl]cyclohexane-1-carboxamide
Figure imgf000241_0001
Step 1: Preparation of 6-ethoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000241_0002
To a solution of nicotinaldehyde (1.07 g, 10 mmol), 6-ethoxybenzo[d]thiazol-2-amine (1.94 g, 10 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.90 g, 50 mmol) in ethanol (100 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-80% ethyl acetate in hexanes to offer 6-ethyl-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (0.430 g, 1.5 mmol, 15 %) as a white powder; LCMS (ESI) m/z: 286.2 [M+H]+. Step 2: Preparation of N-(6-ethoxy-1,3-benzothiazol-2-yl)-1-fluoro-N-[(pyridin-3-yl)methyl]cyclohexane-1- carboxamide
Figure imgf000241_0003
To a reaction vial, a mixture of 6-ethoxy-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (100 mg, 0.35 mmol), 1-fluorocyclohexane-1-carboxylic acid (51.0 mg, 0.35 mmol), N,N-diisopropylethylamine (91.0 mg, 0.70 mmol) and N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (200 mg, 0.52 mmol) in tetrahydrofuran (10 mL) was stirred at 70 °C for 16 hours. The product was indicated present via UPLC analysis. The mixture was poured into water (20 mL) and extracted with 3x 10 mL portions of ethyl acetate. The organic layers were pooled, dried over sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by prep-HPLC to give N-(6-ethoxybenzo[d]thiazol-2-yl)-1-fluoro-N-(pyridin-3- ylmethyl)cyclohexane-1-carboxamide (6.80 mg, 0.016 mmol, 4.7 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 1.34 (t, J = 6.8 Hz, 3H), 1.46-1.66 (m, 6H), 1.93-2.16 (m, 4H), 4.06 (q, J = 7.2 Hz, 2H), 5.58 (s, 2H), 6.94 (dd, J = 8.8, 2.4 Hz, 1H), 7.32 (q, J = 4.4 Hz, 1H), 7.56-7.62 (m, 3H), 8.13 (s, 1H), 8.43 (d, J = 4.4 Hz, 1H), 8.47 (s, 1H); LCMS (ESI) m/z: 414.1 [M+H]+. Example 196. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(3-fluorophenoxy)-N-[(1,3-thiazol- 5-yl)methyl]propenamide
Figure imgf000242_0001
Step 1: Preparation of 5-fluoro-N-(thiazol-5-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000242_0002
To a solution of thiazole-5-carbaldehyde (271 mg, 2.4 mmol), 5-fluorobenzo[d]thiazol-2-amine (400 mg, 2.4 mmol) in toluene (15 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (454 mg, 12 mmol) in ethanol (15 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 5-fluoro-N-(thiazol-5- ylmethyl)benzo[d]thiazol-2-amine (280 mg, 1.1 mmol, 44 %) as a yellow solid. LCMS (ESI) m/z: 266.0 [M+H]+. Step 2: Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(3-fluorophenoxy)-N-[(1,3-thiazol-5- yl)methyl]propanamide
Figure imgf000242_0003
To a solution of 3-(3-fluorophenoxy)propanoic acid (188 mg, 1.0 mmol) in dichloromethane (10 mL) were added 1-hydroxybenzotriazole (138 mg, 1.0 mmol) and 1,3-dicyclohexylcarbodiimide (210 mg, 1.0 mmol). The mixture was stirred for 2 hours. The resulting solution was cooled to 0 °C, and then a solution of 5-fluoro-N-(thiazol-5-ylmethyl)benzo[d]thiazol-2-amine (180 mg, 0.68 mmol) and pyridine (80.0 mg, 1.0 mmol) in N,N-dimethylformamide (10 mL) was added. The reaction mixture was stirred at 40 °C for 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product N-(5-fluorobenzo[d]thiazol-2-yl)-3-(3-fluorophenoxy)-N- (thiazol-5-ylmethyl)propanamide (17.3 mg, 0.040 mmol, 5.9 %) was afforded as a white solid. 1H NMR (500 MHz, Dimethylsulfoxide-d6) δ 9.07 (d, J = 0.5 Hz, 1H), 8.17 (s, 1H), 8.12 (dd, J = 8.8, 5.5 Hz, 1H), 7.77 (dd, J = 9.9, 2.5 Hz, 1H), 7.51 – 7.23 (m, 2H), 7.05 – 6.70 (m, 3H), 5.85 (s, 2H), 4.41 (t, J = 5.8 Hz, 2H), 3.52 – 3.45 (m, 2H); LCMS (ESI) m/z: 432.0 [M+H]+. Example 197. Preparation of 4-fluoro-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]benzamide
Figure imgf000243_0001
Step 1: Preparation of 4-fluoro-N-(6-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)benzamide
Figure imgf000243_0002
In a reaction vial, a mixture of 6-fluoro-N-((1-trityl-1H-imidazol-4-yl)methyl)benzo[d]thiazol-2- amine (300 mg, 0.60 mmol), 4-fluorobenzoic acid (86.0 mg, 0.60 mmol), N,N-diisopropylethylamine (158 mg, 1.2 mmol) and N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (349 mg, 0.90 mmol) in tetrahydrofuran (20 mL) was stirred at 70 °C for 16 hours. The product was indicated present via UPLC analysis. The mixture was poured into water (20 mL) and extracted with 3x 15 mL portions of ethyl acetate. The organic layers were pooled, washed with brine (10 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-40% ethyl acetate in petroleum ether. Product 4-fluoro-N-(6-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H- imidazol-4-yl)methyl)benzamide (195 mg, 0.32 mmol, 54 %) was afforded as a white solid. Step 2: Preparation of 4-fluoro-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]benzamide
Figure imgf000243_0003
To a solution of 4-fluoro-N-(6-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4- yl)methyl)benzamide (195 mg, 0.32 mmol) in dichloromethane (8 mL) was added trifluoroacetic acid (4 mL). The reaction was stirred at room temperature for 3 hours. The product was indicated present via UPLC analysis. The mixture was concentrated under reduced pressure and the residue was quenched with a 7 M solution of ammonia in methanol until pH 7, then extracted with 3x 10 mL portions of ethyl acetate. The organic layers were pooled, dried over sodium sulfate, filtered, concentrated under reduced pressure, and purified by prep-HPLC to give 4-fluoro-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]benzamide (53.0 mg, 0.14 mmol, 45 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 11.91 (s, 1H), 7.81-7.93 (m, 4H), 7.51 (s, 1H), 7.27-7.36 (m, 3H), 6.96 (s, 1H), 5.30 (d, J = 17.2 Hz, 2H); LCMS (ESI) m/z: 371.4 [M+H]+. Example 198. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperidine-1- carboxamide
Figure imgf000244_0001
Step 1: Preparation of 6-fluoro-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000244_0002
In a reaction vial, a solution of pyridine-3-carbaldehyde (126 mg, 1.2 mmol) and 6-fluoro-1,3- benzothiazol-2-amine (200 mg, 1.2 mmol) in dry toluene (12 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry methanol (12 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-fluoro-N-[(pyridin-3- yl)methyl]-1,3-benzothiazol-2-amine (274 mg, 1.1 mmol, 90 %) was afforded as a white solid.1H NMR (300 MHz, Methanol-d4) δ 8.59 (s, 1H), 8.51 – 8.39 (m, 1H), 7.89 (d, J = 8.0 Hz, 1H), 7.54 – 7.27 (m, 3H), 7.01 (tdd, J = 9.0, 2.8, 1.3 Hz, 1H), 4.68 (s, 2H); LCMS (ESI) m/z: 260.0 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperidine-1-carboxamide
Figure imgf000244_0003
In a reaction vial, 6-fluoro-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (274 mg, 1.1 mmol) was dissolved in dichloromethane (52 mL) and cooled to 0 °C. To this solution, pyridine (0.52 mL, 6.5 mmol) and triphosgene (155 mg, 0.53 mmol) were added and the reaction was stirred at 0 °C. After 1 hour, piperidine (97.9 mg, 1.2 mmol) was added at 0 °C and the reaction mixture was stirred at that temperature for 1 additional hour. The product was indicated via UPLC analysis. The reaction mixture was washed with 2x 10 mL deionized water and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product N-(6-fluoro-1,3-benzothiazol-2-yl)- N-[(pyridin-3-yl)methyl]piperidine-1-carboxamide (211 mg, 0.57 mmol, 54 %) was isolated as a yellow oil. 1H NMR (300 MHz, Chloroform-d) δ 8.73 (s, 1H), 8.51 (d, J = 4.3 Hz, 1H), 7.87 (dt, J = 7.9, 2.0 Hz, 1H), 7.63 (dd, J = 8.9, 4.7 Hz, 1H), 7.34 (dd, J = 8.1, 2.6 Hz, 1H), 7.27 (dd, J = 7.9, 4.9 Hz, 1H), 7.08 (td, J = 9.0, 2.6 Hz, 1H), 5.20 (s, 2H), 3.37 (t, J = 5.3 Hz, 4H), 1.67 – 1.40 (m, 6H); LCMS (ESI) m/z: 371.1 [M+H]+. Example 199. Preparation of methyl 4-[(6-methoxy-1,3-benzothiazol-2-yl)[(1,3-thiazol-5- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000245_0001
Step 1: Preparation of 6-methoxy-N-(thiazol-5-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000245_0002
To a solution of 6-methoxybenzo[d]thiazol-2-amine (200 mg, 1.1 mmol) and thiazole-5- carbaldehyde (150 mg, 1.3 mmol) in toluene (10 mL) was added dried 4 Å molecular sieves. The mixture was heated to 120 °C and stirred for 16 hours. Then a solution of sodium borohydride (170 mg, 4.4 mmol) in methanol (10 mL) was added. The reaction was stirred at room temperature for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-10% methanol in dichloromethane. Product 6-methoxy-N-(thiazol-5-ylmethyl)benzo[d]thiazol-2-amine (190 mg, 0.68 mmol, 68 %) was afforded as a white solid. LCMS (ESI) m/z: 278.0 [M+H]+. Step 2: Preparation of tert-butyl 4-((6-methoxybenzo[d]thiazol-2-yl)(thiazol-5- ylmethyl)carbamoyl)piperidine-1-carboxylate
Figure imgf000245_0003
In a reaction vial, a mixture of 6-methoxy-N-(thiazol-5-ylmethyl)benzo[d]thiazol-2-amine (200 mg, 0.72 mmol), 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (250 mg, 1.1 mmol), N,N- diisopropylethylamine (280 mg, 2.2 mmol) and N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (410 mg, 1.1 mmol) in dichloromethane (10 mL) was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. The mixture was poured into water (20 mL) and extracted with 3x 15 mL portions of ethyl acetate. The organic layers were pooled, washed with brine (10 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-25% ethyl acetate in petroleum ether. Product 4- fluoro-N-(6-fluorobenzo[d]thiazol-2-yl)-N-((1-trityl-1H-imidazol-4-yl)methyl)benzamide (210 mg, 0.43 mmol, 60 %) was afforded as a yellow solid. LCMS (ESI) m/z: 489.0 [M+H]+. Step 3: Preparation of N-(6-methoxybenzo[d]thiazol-2-yl)-N-(thiazol-5-ylmethyl)piperidine-4-carboxamide
Figure imgf000246_0001
In a reaction vial, tert-butyl 4-((6-methoxybenzo[d]thiazol-2-yl)(thiazol-5- ylmethyl)carbamoyl)piperidine-1-carboxylate (260 mg, 0.53 mmol) was dissolved in dichloromethane (10 mL). A 4 M solution of hydrochloric acid in dioxane (5 mL) was added to the solution, and the reaction was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The mixture was concentrated under reduced pressure to afford crude N-(6-methoxybenzo[d]thiazol-2-yl)-N- (thiazol-5-ylmethyl)piperidine-4-carboxamide (120 mg, 0.31 mmol, 68 %) as yellow solid, which was used directly in next step without further purification; LCMS (ESI) m/z: 389.0 [M+H]+. Step 4: Preparation of methyl 4-[(6-methoxy-1,3-benzothiazol-2-yl)[(1,3-thiazol-5- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000246_0002
To a solution of N-(6-methoxybenzo[d]thiazol-2-yl)-N-(thiazol-5-ylmethyl)piperidine-4- carboxamide (110 mg, 0.28 mmol) in dichloromethane (3 mL) was added triethylamine (72.0 mg, 0.56 mmol) and methyl carbonochloridate (52.0 mg, 0.56 mmol). The reaction mixture was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The mixture was diluted with water (3 mL) and extracted with dichloromethane (3x 15 mL). The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile/0.1% ammonium bicarbonate) to afford the desire compound methyl 4-((6-methoxybenzo[d]thiazol-2-yl)(thiazol-5-ylmethyl)carbamoyl)piperidine-1-carboxylate (23.6 mg, 0.058 mmol, 20 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.99 (s, 1H), 8.13 (s, 1H), 7.76 (d, J = 8.9 Hz, 1H), 7.59 (d, J = 2.5 Hz, 1H), 7.07 (dd, J = 8.8, 2.6 Hz, 1H), 5.79 (s, 2H), 4.02 – 3.98 (m, 2H), 3.82 (s, 3H), 3.61 (s, 3H), 3.32 – 3.28 (m, 1H), 2.98 – 2.94 (m, 2H), 1.73 – 1.69 (m, 2H), 1.59 – 1.42 (m, 2H); LCMS (ESI) m/z: 447.1 [M+H]+. Example 200. Preparation of 4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5- yl)methyl]benzamide
Figure imgf000246_0003
Step 1: Preparation of 6-fluoro-N-(thiazol-5-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000247_0001
To a solution of 6-fluorobenzo[d]thiazol-2-amine (400 mg, 2.4 mmol) and thiazole-5-carbaldehyde (271 mg, 2.4 mmol) in toluene (15 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (454 mg, 12 mmol) in ethanol (15 mL) was added. The reaction was stirred at 110 °C and stirred for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane. Product 6-fluoro-N-(thiazol-5-ylmethyl)benzo[d]thiazol-2-amine (280 mg, 1.1 mmol, 44 %) was afforded as a yellow solid. LCMS (ESI) m/z: 266.0 [M+H]+. Step 2: Preparation of 4-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5-yl)methyl]benzamide
Figure imgf000247_0002
To a solution of 4-cyanobenzoic acid (182 mg, 1.2 mmol) in dichloromethane (10 mL) were added 1-hydroxybenzotriazole (167 mg, 1.2 mmol) and 1,3-dicyclohexylcarbodiimide (255 mg, 1.2 mmol). The reaction mixture was stirred at room temperature for 2 hours. The resulting solution was cooled to 0 °C, and then a solution of 6-fluoro-N-(thiazol-5-ylmethyl)benzo[d]thiazol-2-amine (220 mg, 0.83 mmol) in pyridine (98.0 mg, 1.2 mmol) and N,N-dimethylformamide (10 mL) was added. The reaction mixture was stirred at 45°C for 17 hours. The product was indicated present via UPLC analysis. Water (20 mL) was added, and the organic layers were extracted with 3x 20 mL washes of ethyl acetate. The organic layers were pooled and concentrated under reduced pressure, and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product 4-cyano-N-(6- fluorobenzo[d]thiazol-2-yl)-N-(thiazol-5-ylmethyl)benzamide (80.3 mg, 0.20 mmol, 25 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.96 (s, 1H), 8.06 (d, J = 8.3 Hz, 2H), 8.00 (dd, J = 8.7, 2.7 Hz, 1H), 7.93 (dd, J = 8.9, 4.8 Hz, 1H), 7.82 (t, J = 13.4 Hz, 2H), 7.51 (s, 1H), 7.38 (td, J = 9.1, 2.7 Hz, 1H), 5.51 (s, 2H); LCMS (ESI) m/z: 394.9 [M+H]+. Example 201. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]pyridine-4-carboxamide
Figure imgf000247_0003
Step 1: Preparation of 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000248_0001
To a solution of 1H-imidazole-4-carbaldehyde (2.50 g, 27 mmol), 6-fluorobenzo[d]thiazol-2-amine (3.00 g, 18 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.30 g, 36 mmol) in methanol (50 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3- benzothiazol-2-amine (2.50 g, 10 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 249.1 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]pyridine-4- carboxamide
Figure imgf000248_0002
To a solution of N-((1H-imidazol-4-yl)methyl)-6-fluorobenzo[d]thiazol-2-amine (100 mg, 0.40 mmol) in tetrahydrofuran (5 mL) was added isonicotinic acid (60.0 mg, 0.48 mmol), N-[(dimethylamino)- 1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (230 mg, 0.60 mmol) and N,N-diisopropylethylamine (104 mg, 0.80 mmol). The reaction mixture was stirred at 70 °C for 16 hours. The product was indicated present via UPLC analysis. Water (20 mL) was added and the reaction was extracted with 2x 20 mL washes of ethyl acetate. The organic layers were pooled and concentrated under reduced pressure. The mixture was purified prep-HPLC to afford N-(6- fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]pyridine-4-carboxamide (36.4 mg, 0.10 mmol, 26 % yield) as a white solid.1H NMR (400 MHz, Methanol-d4) δ 8.66 (dd, J = 4.5, 1.6 Hz, 2H), 7.80 (dd, J = 8.9, 4.6 Hz, 1H), 7.67 (dd, J = 8.4, 2.6 Hz, 1H), 7.59 – 7.52 (m, 2H), 7.22 (td, J = 9.1, 2.7 Hz, 1H), 6.86 (s, 1H), 5.37 (s, 2H); LCMS (ESI) m/z: 354.0 [M+H]+. Example 202. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(propan-2-yloxy)-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000248_0003
Step 1: Preparation of 5-fluoro-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000248_0004
In a round bottom flask equipped with a stir bar, a solution of pyridine-3-carbaldehyde (1.38 mL, 15 mmol) and 5-fluoro-1,3-benzothiazol-2-amine (2.50 g, 15 mmol) in dry toluene (74 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen overnight. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (2.79 g, 74 mmol) in dry ethyl alcohol (74 mL). The reaction mixture was stirred at 70 °C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with ethyl acetate. The filtrate was concentrated then diluted with dichloromethane and filtered again. The filtrate was purified via flash column chromatography eluting with 0-100% hexanes/dichloromethane. Product 5-fluoro-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (1.26 g, 4.87 mmol, 33 %) was afforded as a white powder.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 8.95 – 8.33 (m, 2H), 7.91 – 7.59 (m, 2H), 7.29 (ddd, J = 47.6, 9.2, 3.7 Hz, 2H), 6.88 (td, J = 9.1, 2.6 Hz, 1H), 4.63 (d, J = 4.7 Hz, 2H), 3.36 (s, 1H); LCMS (ESI) m/z: 261.0 [M+H]+. Step 2: N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(propan-2-yloxy)-N-[(pyridin-3-yl)methyl]propenamide
Figure imgf000249_0001
In a reaction vial equipped with a stir bar, 5-fluoro-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2- amine (250 mg, 0.96 mmol) was dissolved in dichloromethane (5 mL). To the reaction solution was added 3-(propan-2-yloxy)propanoic acid (506 mg, 3.8 mmol) and 50% propylphosphonic anhydride in ethyl acetate (3.64 mL, 4.8 mmol). The reaction mixture was cooled to 0 °C and triethylamine (0.010 mL, 9.6 mmol) was added. The reaction was stirred from 0 °C to room temperature over the weekend. The product was indicated present via UPLC analysis. The reaction solution was diluted with dichloromethane and washed with water (3x 15 mL). The organic layers were pooled and dried over anhydrous sodium sulfate. The organic layer was filtered and concentrated. The product was purified via flash column chromatography eluting with 0-100% ethyl acetate in hexanes through a 24 g column of silica gel. Product N-(5-fluoro-1,3-benzothiazol-2-yl)-3-(propan-2-yloxy)-N-[(pyridin-3-yl)methyl]propenamide (57.2 mg, 0.15 mmol, 16 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.60 (dd, J = 17.3, 3.4 Hz, 2H), 7.75 (dd, J = 8.7, 5.2 Hz, 1H), 7.62 (d, J = 8.0 Hz, 1H), 7.47 (dd, J = 9.7, 2.4 Hz, 1H), 7.38 – 7.20 (m, 2H), 7.09 (td, J = 8.8, 2.4 Hz, 1H), 5.67 (s, 2H), 3.84 (t, J = 6.4 Hz, 2H), 3.62 (p, J = 6.1 Hz, 1H), 2.88 (t, J = 6.4 Hz, 2H), 1.15 (d, J = 6.1 Hz, 6H); LCMS (ESI) m/z: 374.1 [M+H]+. Example 203. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-4-yl)methyl]benzamide
Figure imgf000249_0002
Step 1: Preparation of 6-fluoro-N-[(pyridin-4-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000250_0001
In a reaction vial, a solution of 6-fluoro-1,3-benzothiazol-2-amine (200 mg, 1.2 mmol) and pyridine-4-carbaldehyde (126 mg, 1.2 mmol) in dry toluene (12 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry ethyl alcohol (12 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-fluoro-N-[(pyridin- 4-yl)methyl]-1,3-benzothiazol-2-amine (228 mg, 0.88 mmol, 75 %) was afforded as a yellow-orange solid. 1H NMR (300 MHz, Methanol-d4) δ 8.59 – 8.28 (m, 2H), 7.45 (d, J = 5.5 Hz, 2H), 7.41 – 7.27 (m, 2H), 7.01 (td, J = 9.2, 2.6 Hz, 1H), 4.69 (d, J = 10.4 Hz, 2H); LCMS (ESI) m/z: 260.0 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-4-yl)methyl]benzamide
Figure imgf000250_0002
In a reaction vial, 6-fluoro-N-[(pyridin-4-yl)methyl]-1,3-benzothiazol-2-amine (229 mg, 0.88 mmol) and N,N-diisopropylethylamine (0.46 mL, 2.6 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. benzoyl chloride (184 mg, 1.3 mmol) was added dropwise with stirring and the reaction was stirred for 16 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyridin-4-yl)methyl]benzamide (75.9 mg, 0.21 mmol, 24 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.51 (d, J = 5.7 Hz, 2H), 7.69 (dd, J = 8.9, 4.7 Hz, 1H), 7.57 – 7.45 (m, 2H), 7.41 (d, J = 4.4 Hz, 4H), 7.15 (td, J = 9.0, 2.6 Hz, 1H), 7.05 (d, J = 5.1 Hz, 2H), 5.48 (s, 2H); LCMS (ESI) m/z: 364.2 [M+H]+. Example 204. Preparation of 1-acetyl-N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3- yl)methyl]piperidine-4-carboxamide
Figure imgf000250_0003
Step 1: Preparation of 6-methoxy-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000251_0001
In a reaction vial, a solution of pyridine-3-carbaldehyde (117 mg, 1.1 mmol) and 6-methoxy-1,3- benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (95.7 mg, 2.5 mmol) in dry methanol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-methoxy-N-[(pyridin-3- yl)methyl]-1,3-benzothiazol-2-amine (248 mg, 0.91 mmol, 83 %) was afforded as a yellow solid.1H NMR (300 MHz, Methanol-d4) δ 8.70 – 8.46 (m, 1H), 8.54 – 8.37 (m, 1H), 7.98 – 7.79 (m, 1H), 7.43 (dd, J = 7.9, 5.0 Hz, 1H), 7.33 (dd, J = 8.9, 1.1 Hz, 1H), 7.20 (dd, J = 2.7, 1.1 Hz, 1H), 6.87 (ddd, J = 8.8, 2.7, 1.2 Hz, 1H), 4.67 (s, 2H), 3.79 (d, J = 1.3 Hz, 3H); LCMS (ESI) m/z: 272.0 [M+H]+. Step 2: Preparation of 1-acetyl-N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperidine-4- carboxamide
Figure imgf000251_0002
In a reaction vial, 6-methoxy-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (258 mg, 0.95 mmol), 1-acetylpiperidine-4-carboxylic acid (323 mg, 1.9 mmol), and triethylamine (0.66 mL, 4.7 mmol) were dissolved in dichloromethane (5 mL) with stirring. The reaction was cooled to 0 °C, and 50% propylphosphonic anhydride in ethyl acetate (1.8 mL, 2.4 mmol) was added. The reaction was warmed to room temperature and stirred for 3 days. The reaction was diluted with 10 mL deionized water and washed with 2x 10 mL portions of dichloromethane. The organic layers were separated, pooled, and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 1-acetyl-N-(6- methoxy-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperidine-4-carboxamide (156 mg, 0.37 mmol, 39 %) was afforded as a yellow solid.1H NMR (300 MHz, Methanol-d4) δ 8.56 (s, 1H), 8.47 (d, J = 4.9 Hz, 1H), 7.77 (d, J = 8.0 Hz, 1H), 7.63 (d, J = 8.9 Hz, 1H), 7.48 – 7.35 (m, 2H), 7.02 (d, J = 8.9 Hz, 1H), 5.72 (s, 2H), 4.50 (d, J = 13.3 Hz, 1H), 4.03 – 3.90 (m, 1H), 3.85 (d, J = 1.3 Hz, 3H), 3.11 (d, J = 21.8 Hz, 2H), 2.67 (t, J = 12.8 Hz, 1H), 2.09 (d, J = 1.2 Hz, 3H), 1.88 – 1.70 (m, 3H); LCMS (ESI) m/z: 425.2 [M+H]+. Example 205. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-1-methyl-2-oxo-N-[(pyridin-3- yl)methyl]piperidine-4-carboxamide
Figure imgf000252_0001
Step 1: Preparation of 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000252_0002
In a round bottom flask equipped with a stir bar, a solution of pyridine-3-carbaldehyde (0.63 mL, 6.7 mmol) and 6-ethyl-1,3-benzothiazol-2-amine (1.20 g, 6.7 mmol) in dry toluene (34 mL) was stirred at 105°C with activated 4 Å molecular sieves under nitrogen overnight. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (1.27 g, 34 mmol) in dry ethyl alcohol (34 mL). The reaction mixture was stirred at 70 °C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with ethyl acetate. The filtrate was concentrated then diluted with dichloromethane and filtered again. The filtrate was purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane through a 40 g column of silica gel. Product 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3- benzothiazol-2-amine (0.920 g, 3.4 mmol, 51 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.72 – 8.50 (m, 2H), 7.76 (dddd, J = 7.9, 2.4, 1.7, 0.8 Hz, 1H), 7.50 – 7.37 (m, 2H), 7.35 – 7.22 (m, 1H), 7.14 (dt, J = 8.3, 1.2 Hz, 1H), 6.24 (s, 1H), 4.68 (s, 2H), 2.70 (q, J = 7.6 Hz, 2H), 1.26 (td, J = 7.6, 0.7 Hz, 3H); LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of 1-methyl-2-oxopiperidine-4-carbonyl chloride
Figure imgf000252_0003
In a reaction vial, 1-methyl-2-oxopiperidine-4-carboxylic acid (150 mg, 0.95 mmol) was dissolved in thionyl chloride (1.37 mL, 19 mmol) and stirred at room temperature for 3 hours. The reaction was concentrated under reduced pressure. Crude product was indicated pure enough to continue to next step via NMR. Assumed 100% yield. Product 1-methyl-2-oxopiperidine-4-carbonyl chloride (167 mg, 0.95 mmol, 100 %) was afforded as a yellow oil.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 3.35 – 3.15 (m, 1H), 2.78 (d, J = 1.6 Hz, 2H), 2.50 (h, J = 1.8 Hz, 6H), 2.33 (t, J = 7.1 Hz, 1H). Step 3: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-1-methyl-2-oxo-N-[(pyridin-3-yl)methyl]piperidine- 4-carboxamide In a reaction vial equipped with a magnetic stir bar, 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3- benzothiazol-2-amine (185 mg, 0.69 mmol) was dissolved in tetrahydrofuran (3.5 mL) and cooled to 0 °C. To the solution was added triethylamine (0.24 mL, 1.7 mmol) then 1-methyl-2-oxopiperidine-4-carbonyl chloride (179 mg, 1.0 mmol) was added, slowly. The reaction was stirred from 0 °C to room temperature overnight. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane and washed with water (3x). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Crude product was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane through 24 g of silica gel. Product N-(6-ethyl-1,3-benzothiazol-2- yl)-1-methyl-2-oxo-N-[(pyridin-3-yl)methyl]piperidine-4-carboxamide (35.1 mg, 0.086 mmol, 13 %) was afforded as a yellow oil.1H NMR (300 MHz, Chloroform-d) δ 8.67 – 8.49 (m, 2H), 7.84 – 7.53 (m, 3H), 7.38 – 7.17 (m, 3H), 3.27 (ddt, J = 28.1, 12.5, 6.5 Hz, 3H), 2.95 (s, 3H), 2.89 – 2.49 (m, 4H), 2.00 (d, J = 52.8 Hz, 2H), 1.31 (td, J = 7.6, 0.9 Hz, 4H); LCMS (ESI) m/z: 409.1 [M+H]+. Example 206. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5- yl)methyl]piperidine-1-carboxamide
Figure imgf000253_0001
Step 1: Preparation of 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000253_0002
In a reaction vial, a solution of 1H-imidazole-5-carbaldehyde (113 mg, 1.2 mmol) and 6-fluoro-1,3- benzothiazol-2-amine (200 mg, 1.2 mmol) in dry toluene (12 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry methanol (12 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-20% methanol in ethyl acetate. Product 6-fluoro-N-[(1H-imidazol-5- yl)methyl]-1,3-benzothiazol-2-amine (245 mg, 0.99 mmol, 84 %) was afforded as a pink-brown solid.1H NMR (300 MHz, Methanol-d4) δ 7.66 (d, J = 1.0 Hz, 1H), 7.46 – 7.32 (m, 2H), 7.08 (s, 1H), 7.01 (td, J = 9.1, 2.6 Hz, 1H), 4.57 (s, 2H); LCMS (ESI) m/z: 249.0 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]piperidine-1- carboxamide
Figure imgf000253_0003
In a reaction vial, 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine (246 mg, 0.99 mmol) was dissolved in dichloromethane (49 mL) and cooled to 0 °C. To this solution, pyridine (0.49 mL, 6.1 mmol) and triphosgene (145 mg, 0.49 mmol) were added and the reaction was stirred at 0 °C. After 1 hour, piperidine (91.9 mg, 1.1 mmol) was added at 0 °C and the reaction mixture was stirred at that temperature for 1 additional hour. The product was indicated via UPLC analysis. The reaction mixture was washed with 2x 10 mL deionized water and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product N-(6-fluoro-1,3-benzothiazol-2-yl)- N-[(1H-imidazol-5-yl)methyl]piperidine-1-carboxamide (45.2 mg, 0.13 mmol, 13 %) was isolated as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 7.84 (d, J = 1.3 Hz, 1H), 7.47 (dd, J = 8.8, 4.7 Hz, 1H), 7.31 – 7.26 (m, 1H), 7.19 (d, J = 1.2 Hz, 1H), 7.01 (td, J = 9.0, 2.7 Hz, 1H), 5.60 (dd, J = 5.9, 1.9 Hz, 1H), 4.59 (s, 2H), 4.00 – 3.86 (m, 2H), 3.52 (t, J = 5.2 Hz, 5H), 2.01 – 1.76 (m, 3H); LCMS (ESI) m/z: 360.1 [M+H]+. Example 207. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(2-methylpyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000254_0001
Step 1: Preparation of 6-ethyl-N-[(2-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000254_0002
In a reaction vial, a solution of 2-methylpyridine-3-carbaldehyde (135 mg, 1.1 mmol) and 6-ethyl- 1,3-benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry methanol (11 mL). The resultant reaction mixture was refluxed at 100 °C for 1 hour and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-ethyl-N-[(2-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (214 mg, 0.76 mmol, 68 %) was afforded as a yellow solid.1H NMR (300 MHz, Methanol-d4) δ 8.31 (t, J = 5.9 Hz, 1H), 7.80 (dd, J = 12.9, 7.7 Hz, 1H), 7.44 (s, 1H), 7.33 (d, J = 8.2 Hz, 1H), 7.27 – 7.19 (m, 1H), 7.11 (dd, J = 8.3, 1.8 Hz, 1H), 4.66 (d, J = 1.9 Hz, 2H), 3.35 (d, J = 1.0 Hz, 3H), 2.67 (q, J = 7.5 Hz, 2H), 1.24 (td, J = 7.6, 1.0 Hz, 3H); LCMS (ESI) m/z: 284.0 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(2-methylpyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000255_0001
In a reaction vial, 6-ethyl-N-[(2-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (214 mg, 0.76 mmol) and N,N-diisopropylethylamine (0.39 mL, 2.3 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (165 mg, 1.1 mmol) was added dropwise with stirring and the reaction was stirred for 16 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(2-methylpyridin-3- yl)methyl]cyclohexanecarboxamide (102 mg, 0.26 mmol, 34 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.49 – 8.29 (m, 1H), 7.69 – 7.59 (m, 2H), 7.26 – 7.20 (m, 1H), 7.15 (d, J = 7.6 Hz, 1H), 7.03 (dd, J = 7.8, 4.8 Hz, 1H), 5.54 (s, 2H), 2.75 (q, J = 7.7 Hz, 2H), 2.70 (s, 3H), 2.45 (s, 1H), 1.87 – 1.72 (m, 4H), 1.64 (p, J = 11.3, 9.9 Hz, 4H), 1.34 – 1.22 (m, 3H), 1.24 – 1.08 (m, 2H); LCMS (ESI) m/z: 394.1 [M+H]+. Example 208. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]-2- azaspiro[3.3]heptane-2-carboxamide
Figure imgf000255_0002
Step 1: Preparation of 6-ethyl-N-(pyrimidin-5-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000255_0003
To a solution of pyrimidine-5-carbaldehyde (364 mg, 3.4 mmol) and 6-ethylbenzo[d]thiazol-2- amine (600 mg, 3.4 mmol) in toluene (100 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (640 mg, 17 mmol) in methanol (60 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-10% methanol in dichloromethane to offer 6-ethyl-N- (pyrimidin-5-ylmethyl)benzo[d]thiazol-2-amine (160 mg, 0.59 mmol, 18 %) as a white solid. LCMS (ESI) m/z: 271.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]-2-azaspiro[3.3]heptane- 2-carboxamide
Figure imgf000256_0001
To a solution of 2-azaspiro[3.3]heptane (115 mg, 1.2 mmol) in tetrahydrofuran (15 mL) was added triethylamine (180 mg, 1.8 mmol), and triphosgene (32.0 mg, 0.29 mmol). The mixture was stirred at 0 °C for 15 minutes, then 6-ethyl-N-(pyrimidin-5-ylmethyl)benzo[d]thiazol-2-amine (160 mg, 0.59 mmol) was added and the reaction was warmed to room temperature with stirring over 16 hours. The product was indicated present via UPLC analysis. The reaction was quenched by ice water (20 mL), then extracted with 3x 20 mL portions of ethyl acetate. The organic layers were pooled, dried over sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by prep-HPLC to afford N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyrimidin-5-ylmethyl)-2-azaspiro[3.3]heptane-2-carboxamide (6.60 mg, 0.017 mmol, 2.8%) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 9.07 (s, 1H), 8.72 (s, 2H), 7.73 (s, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.19 (dd, J = 8.4, 1.6 Hz, 1H), 5.28 (s, 2H), 4.03 (s, 4H), 2.67 (q, J = 7.2 Hz, 2H), 2.10 (t, J = 7.2 Hz, 4H), 1.67-1.75 (m, 2H), 1.20 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 394.0 [M+H]+. Example 209. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]pyridine-3-carboxamide
Figure imgf000256_0002
Step 1: Preparation of 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000256_0003
To a solution of 1H-imidazole-4-carbaldehyde (2.50 g, 27 mmol), 6-fluorobenzo[d]thiazol-2-amine (3.00 g, 18 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.30 g, 36 mmol) in methanol (50 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3- benzothiazol-2-amine (2.50 g, 10 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 249.1 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]pyridine-3- carboxamide
Figure imgf000256_0004
To a solution of N-((1H-imidazol-4-yl)methyl)-6-fluorobenzo[d]thiazol-2-amine (150 mg, 0.60 mmol) in tetrahydrofuran (10 mL) was added nicotinic acid (89.3 mg, 0.72 mmol), N-[(dimethylamino)-1H- 1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (342 mg, 1.2 mmol) and N,N-diisopropylethylamine (155 mg, 0.90 mmol). The reaction mixture was stirred at 70 °C for 16 hours. The product was indicated present via UPLC analysis. Water (20 mL) was added and the reaction was extracted with 2x 20 mL washes of ethyl acetate. The organic layers were pooled and concentrated under reduced pressure. The mixture was purified prep-HPLC to afford N-(6-fluoro-1,3- benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]pyridine-3-carboxamide (101 mg, 0.29 mmol, 48 % yield) as a white solid.1H NMR (400 MHz, Methanol-d4) δ 8.69 (dd, J = 15.9, 2.6 Hz, 2H), 8.04 (d, J = 7.3 Hz, 1H), 7.79 (s, 1H), 7.66 (dd, J = 8.4, 2.6 Hz, 1H), 7.60 – 7.38 (m, 2H), 7.21 (td, J = 9.0, 2.5 Hz, 1H), 6.88 (s, 1H), 5.41 (s, 2H); LCMS (ESI) m/z: 354.0 [M+H]+. Example 210. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]pyridine-2-carboxamide
Figure imgf000257_0001
Step 1: Preparation of 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000257_0002
To a solution of 1H-imidazole-4-carbaldehyde (2.50 g, 27 mmol), 6-fluorobenzo[d]thiazol-2-amine (3.00 g, 18 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.30 g, 36 mmol) in methanol (50 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3- benzothiazol-2-amine (2.50 g, 10 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 249.1 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]pyridine-2- carboxamide
Figure imgf000257_0003
To a solution of N-((1H-imidazol-4-yl)methyl)-6-fluorobenzo[d]thiazol-2-amine (150 mg, 0.60 mmol) in tetrahydrofuran (10 mL) was added picolinic acid (89.3 mg, 0.72 mmol), N-[(dimethylamino)-1H- 1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (342 mg, 1.2 mmol) and N,N-diisopropylethylamine (155 mg, 0.90 mmol). The reaction mixture was stirred at 70 °C for 16 hours. The product was indicated present via UPLC analysis. Water (20 mL) was added and the reaction was extracted with 2x 20 mL washes of ethyl acetate. The organic layers were pooled and concentrated under reduced pressure. The mixture was purified prep-HPLC to afford N-(6-fluoro-1,3- benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]pyridine-2-carboxamide (128 mg, 0.36 mmol, 60% yield) as a white solid.1H NMR (400 MHz, Methanol-d4) δ 8.65 (d, J = 4.5 Hz, 1H), 7.95 (dd, J = 8.5, 7.1 Hz, 1H), 7.82 (dd, J = 8.8, 4.7 Hz, 1H), 7.76 – 7.62 (m, 2H), 7.54 (dd, J = 19.9, 13.2 Hz, 2H), 7.22 (td, J = 9.1, 2.7 Hz, 1H), 7.22 (td, J = 9.1, 2.7 Hz, 1H), 6.73 (s, 1H), 5.68 (s, 2H); LCMS (ESI) m/z: 354.0 [M+H]+. Example 211. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperazine-1- carboxamide
Figure imgf000258_0001
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyridin-3-yl)methyl]piperazine-1-carboxamide
Figure imgf000258_0002
In a reaction vial, tert-butyl 4-((6-ethylbenzo[d]thiazol-2-yl)(pyridin-3- ylmethyl)carbamoyl)piperazine-1-carboxylate (360 mg, 0.75 mmol) was dissolved in dichloromethane (15 mL). Trifluoroacetic acid (8 mL) was added and the reaction was stirred at room temperature for 3 hours. The product was indicated present via UPLC analysis. The reaction was concentrated under reduced pressure. The residue was re-dissolved in water (20 mL) and extracted with 3x 20 mL portions of ethyl acetate. The organic layers were pooled, dried over sodium sulfate, filtered, and concentrated under reduced pressure. Crude product was purified by prep-HPLC to give N-(6-ethylbenzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)piperazine-1-carboxamide (150 mg, 0.039 mmol, 53 %) as a white solid.1H NMR (500 MHz, Chloroform-d) δ 8.70 (s, 1H), 8.49 (s, 1H), 7.82 (d, J = 8.0 Hz, 1H), 7.63 (d, J = 8.5 Hz, 1H), 7.48 (s, 1H), 7.23 (t, J = 12.5 Hz, 2H), 5.18 (s, 2H), 3.44 (d, J = 4.5 Hz, 4H), 2.86 (s, 4H), 2.72 (q, J = 7.5 Hz, 2H), 1.26 (t, J = 8.0 Hz, 3H); LCMS (ESI) m/z: 382.2 [M+H]+. Example 212. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-1-methyl-N-[(pyridin-3- yl)methyl]piperidine-4-carboxamide
Figure imgf000258_0003
Step 1: Preparation of 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000258_0004
In a round bottom flask equipped with a stir bar, a solution of pyridine-3-carbaldehyde (0.63 mL, 6.7 mmol) and 6-ethyl-1,3-benzothiazol-2-amine (1.20 g, 6.7 mmol) in dry toluene (34 mL) was stirred at 105°C with activated 4 Å molecular sieves under nitrogen overnight. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (1.27 g, 34 mmol) in dry ethyl alcohol (34 mL). The reaction mixture was stirred at 70 °C for 30 minutes and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with ethyl acetate. The filtrate was concentrated then diluted with dichloromethane and filtered again. The filtrate was purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane through a 40 g column of silica gel. Product 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3- benzothiazol-2-amine (0.915 g, 3.4 mmol, 51 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.72 – 8.50 (m, 2H), 7.76 (dddd, J = 7.9, 2.4, 1.7, 0.8 Hz, 1H), 7.50 – 7.37 (m, 2H), 7.35 – 7.22 (m, 1H), 7.14 (dt, J = 8.3, 1.2 Hz, 1H), 6.24 (s, 1H), 4.68 (s, 2H), 2.70 (q, J = 7.6 Hz, 2H), 1.26 (td, J = 7.6, 0.7 Hz, 3H); LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-1-methyl-N-[(pyridin-3-yl)methyl]piperidine-4- carboxamide
Figure imgf000259_0001
In a reaction vial equipped with a magnetic stir bar, 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3- benzothiazol-2-amine (375 mg, 1.4 mmol) was dissolved in tetrahydrofuran (7 mL) and cooled to 0 °C. To the solution was added triethylamine (0.48 mL, 3.5 mmol) then 1-methylpiperidine-4-carbonyl chloride (336 mg, 2.1 mmol) was added, slowly. The reaction was stirred from 0 °C to room temperature overnight. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane and washed with water (3x). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Crude product was purified via flash column chromatography eluting with 0-20% methanol in dichloromethane. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-1-methyl-N-[(pyridin- 3-yl)methyl]piperidine-4-carboxamide (62.1 mg, 0.16 mmol, 11 %) was afforded as pale yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.73 – 8.49 (m, 2H), 7.81 – 7.51 (m, 3H), 7.38 – 7.19 (m, 2H), 5.62 (s, 2H), 3.07 – 2.53 (m, 5H), 2.28 (s, 3H), 2.14 – 1.50 (m, 6H), 1.30 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 395.1 [M+H]+. Example 213. Preparation of N-(5-fluoro-1,3-benzoxazol-2-yl)-N-[(1H-imidazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000259_0002
Step 1: Preparation of 5-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzoxazol-2-amine
Figure imgf000260_0001
In a round bottom flask equipped with a magnetic stir bar, a solution of 1H-imidazole-5- carbaldehyde (179 mg, 1.9 mmol) and 5-fluoro-1,3-benzoxazol-2-amine (300 mg, 2.0 mmol) in dry toluene (10 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen for 24 hours. The reaction mixture was cooled to 60 °C and then in was added of sodium borohydride (171 mg, 4.5 mmol) and dry methanol (10 mL). The reaction mixture was stirred at 70 °C for 1 hour and then cooled to room temperature. The reaction was filtered over a bed of Celite and washed with methanol (2x 20 mL). The filtrate was concentrated then diluted with dichloromethane and re-filtered. The filtrate was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane. Product 5-fluoro-N- [(1H-imidazol-5-yl)methyl]-1,3-benzoxazol-2-amine (77.9 mg, 0.34 mmol, 17 %) was afforded as an orange powder.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 8.41 (d, J = 5.7 Hz, 1H), 7.58 (s, 1H), 7.32 (ddd, J = 8.6, 4.5, 1.4 Hz, 1H), 7.20 – 6.87 (m, 1H), 6.86 – 6.65 (m, 1H), 4.40 (d, J = 5.7 Hz, 1H), 3.34 (d, J = 1.4 Hz, 2H); LCMS (ESI) m/z: 233.0 [M+H]+. Step 2: Preparation of N-(5-fluoro-1,3-benzoxazol-2-yl)-N-[(1H-imidazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000260_0002
In a reaction vial equipped with a magnetic stir bar, 5-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3- benzoxazol-2-amine (77.9 mg, 0.34 mmol) was dissolved in tetrahydrofuran (2 mL) and cooled to 0 °C. To the solution was added triethylamine (0.070 mL, 0.50 mmol) then cyclohexanecarbonyl chloride (0.067 mL, 0.50 mmol) was added, slowly. The reaction was stirred from 0 °C to room temperature overnight. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane and washed with water (3x). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Crude product was purified via flash column chromatography eluting with 0-100% ethyl acetate in hexanes. Product N-(5-fluoro-1,3-benzoxazol-2-yl)-N-[(1H-imidazol-5- yl)methyl]cyclohexanecarboxamide (20.5 mg, 0.060 mmol, 18 %) was afforded as an amber oil.1H NMR (300 MHz, Chloroform-d) δ 7.52 – 7.23 (m, 3H), 7.18 – 6.96 (m, 2H), 5.13 (s, 2H), 3.24 (d, J = 11.3 Hz, 1H), 2.00 – 1.62 (m, 5H), 1.51 (d, J = 12.9 Hz, 2H), 1.40 – 1.11 (m, 4H); LCMS (ESI) m/z: 343.1 [M+H]+. Example 214. Preparation of methyl 4-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyrimidin-5- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000260_0003
Preparation of methyl 4-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyrimidin-5-yl)methyl]carbamoyl]piperidine-1- carboxylate
Figure imgf000261_0001
To a solution of N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyrimidin-5-ylmethyl)piperidine-4-carboxamide (60.0 mg, 0.15 mmol) in dichloromethane (3 mL) was added triethylamine (30.0 mg, 0.30 mmol) and methyl carbonochloridate (16.0 mg, 0.17 mmol). The reaction was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The reaction was diluted with water (3 mL), then extracted with dichloromethane (3x 15 mL). The organic layers were pooled and concentrated under reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N- dimethylformamide before purification. Boston pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile/0.1% ammonium bicarbonate) to afford the desire compound methyl 4-((6- ethylbenzo[d]thiazol-2-yl)(pyrimidin-5-ylmethyl)carbamoyl)piperidine-1-carboxylate (31.5 mg, 0.070 mmol, 48 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 9.11 (s, 1H), 8.79 (s, 2H), 7.81 (s, 1H), 7.64 (d, J = 8.3 Hz, 1H), 7.28 (d, J = 8.5 Hz, 1H), 5.64 (s, 2H), 4.01 – 3.96 (m, 2H), 3.60 (s, 3H), 3.27– 3.20 (m, 1H), 2.90 – 2.85 (m, 2H), 1.79 (d, J = 12.2 Hz, 2H), 1.81 – 1.77 (m, 2H), 1.81 – 1.77 (m, 2H), 1.21 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 440.3 [M+H]+. Example 215. Preparation of tert-butyl 4-{[(pyrimidin-5-yl)methyl][6-(trifluoromethoxy)-1,3- benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000261_0002
Step 1: Preparation of N-(pyrimidin-5-ylmethyl)-6-(trifluoromethoxy)benzo[d]thiazol-2-amine
Figure imgf000261_0003
To a solution of pyrimidine-5-carbaldehyde (92.0 mg, 0.85 mmol) and 6- (trifluoromethoxy)benzo[d]thiazol-2-amine (200 mg, 0.85 mmol) in toluene (8 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (161 mg, 4.3 mmol) in ethanol (8 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer N-(pyrimidin-5-ylmethyl)-6-(trifluoromethoxy)benzo[d]thiazol-2-amine (20.0 mg, 0.060 mmol, 7.2%) as a white solid. LCMS (ESI) m/z: 327.0 [M+H]+. Step 2: Preparation of tert-butyl 4-{[(pyrimidin-5-yl)methyl][6-(trifluoromethoxy)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000262_0001
To a solution of N-(pyrimidin-5-ylmethyl)-6-(trifluoromethoxy)benzo[d]thiazol-2-amine (20.0 mg, 0.060 mmol) in N,N-dimethylformamide (8 mL) were added 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (27.0 mg, 0.12 mmol) and N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (34.0 mg, 0.090 mmol) and triethylamine (18.0 mg, 0.18 mmol). The mixture was stirred at 45°C for 17 hours. The product was indicated present via UPLC analysis. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid). Product tert-butyl 4-((pyrimidin-5-ylmethyl)(6-(trifluoromethoxy)benzo[d]thiazol-2- yl)carbamoyl)piperidine-1-carboxylate (4.55 mg, 0.0085 mmol, 14.1 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 9.12 (s, 1H), 8.82 (s, 2H), 8.14 (d, J = 1.4 Hz, 1H), 7.83 (d, J = 8.8 Hz, 1H), 7.41 (d, J = 8.7 Hz, 1H), 5.66 (s, 2H), 3.95 (d, J = 12.1 Hz, 2H), 3.24 (d, J = 11.2 Hz, 1H), 2.82 (t, J = 31.9 Hz, 2H), 1.77 (d, J = 12.0 Hz, 2H), 1.63 – 1.44 (m, 2H), 1.38 (d, J = 19.6 Hz, 9H); LCMS (ESI) m/z: 538.2 [M+H]+. Example 216. Preparation of 3-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4- yl)methyl]benzamide
Figure imgf000262_0002
Step 1: Preparation of 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000262_0003
To a solution of 1H-imidazole-4-carbaldehyde (2.50 g, 27 mmol), 6-fluorobenzo[d]thiazol-2-amine (3.00 g, 18 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.30 g, 36 mmol) in methanol (50 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-fluoro-N-[(1H-imidazol-5- yl)methyl]-1,3-benzothiazol-2-amine (2.50 g, 10 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 249.1 [M+H]+. Step 2: Preparation of 3-cyano-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]benzamide
Figure imgf000263_0001
To a solution of N-((1H-imidazol-4-yl)methyl)-6-fluorobenzo[d]thiazol-2-amine (150 mg, 0.60 mmol) in tetrahydrofuran (10 mL) was added 3-cyanobenzoic acid (106 mg, 0.72 mmol), N- [(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (342 mg, 1.2 mmol) and N,N-diisopropylethylamine (155 mg, 0.90 mmol). The reaction mixture was stirred at 70 °C for 16 hours. The product was indicated present via UPLC analysis. Water (20 mL) was added and the reaction was extracted with 2x 20 mL washes of ethyl acetate. The organic layers were pooled and concentrated under reduced pressure. The mixture was purified prep-HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2ml/min and the solvent was acetonitrile/0.01% aqueous ammonium bicarbonate) to afford N-((1H-imidazol-4-yl)methyl)-3-cyano-N-(6- fluorobenzo[d]thiazol-2-yl)benzamide (110 mg, 0.29 mmol, 49 %) as white solid.1H NMR (400 MHz, Methanol-d4) δ 8.00 – 7.86 (m, 3H), 7.83 (dd, J = 8.6, 4.5 Hz, 1H), 7.73 – 7.63 (m, 2H), 7.60 (d, J = 1.0 Hz, 1H), 7.24 (td, J = 9.0, 2.6 Hz, 1H), 6.90 (s, 1H), 5.40 (s, 2H); LCMS (ESI) m/z: 378.0 [M+H]+. Example 217. Preparation of tert-butyl 4-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyrimidin-5- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000263_0002
Step 1: Preparation of 6-ethyl-N-(pyrimidin-5-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000263_0003
To a solution of pyrimidine-5-carbaldehyde (364 mg, 3.4 mmol) and 6-ethylbenzo[d]thiazol-2- amine (600 mg, 3.4 mmol) in toluene (100 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (640 mg, 17 mmol) in methanol (60 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-10% methanol in dichloromethane to offer 6-ethyl-N- (pyrimidin-5-ylmethyl)benzo[d]thiazol-2-amine (160 mg, 0.59 mmol, 18 %) as a white solid. LCMS (ESI) m/z: 271.1 [M+H]+. Step 2: Preparation of tert-butyl 4-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyrimidin-5- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000264_0001
To a solution of 6-ethyl-N-(pyrimidin-5-ylmethyl)benzo[d]thiazol-2-amine (100 mg, 0.37 mmol) in dichloromethane (10 mL) was added 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (127 mg, 0.55 mmol), N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (211 mg, 0.55 mmol) and N,N-diisopropylethylamine (101 mg, 0.78 mmol). The reaction mixture was stirred at room temperature for 16 hours. The product was indicated present via UPLC analysis. Water (20 mL) was added and the reaction was extracted with 2x 20 mL washes of ethyl acetate. The organic layers were pooled and concentrated under reduced pressure. The mixture was purified prep-HPLC (the crude samples were dissolved in methanol before purification. Boston C18 21*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous ammonium bicarbonate) to give the desired product tert-butyl 4-((6-ethylbenzo[d]thiazol-2-yl)(pyrimidin-5- ylmethyl)carbamoyl)piperidine-1-carboxylate (41.0 mg, 0.085 mmol, 23 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 9.11 (s, 1H), 8.79 (s, 2H), 7.81 (s, 1H), 7.66 – 7.63 (m, 1H), 7.29 – 7.26 (m, 1H), 5.64 (s, 2H), 3.98 – 3.92 (m, 2H), 3.28 – 3.17 (m, 1H), 2.83 – 2.66 (m, 4H), 1.77 – 1.74 (m, 2H), 1.57 – 1.46 (m, 2H), 1.40 (s, 9H), 1.22 (t, J = 7.5 Hz, 3H); LCMS (ESI) m/z: 482.0 [M+H]+. Example 218. Preparation of N-(6-bromo-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]piperidine-1-carboxamide
Figure imgf000264_0002
Step 1: Preparation of 6-bromo-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000264_0003
In a reaction vial, a solution of pyrimidine-5-carbaldehyde (164 mg, 1.5 mmol) and 6-bromo-1,3- benzothiazol-2-amine (350 mg, 1.5 mmol) in dry toluene (15 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (132 mg, 3.5 mmol) in dry ethyl alcohol (15 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-bromo-N- [(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine (176 mg, 0.55 mmol, 36 %) was afforded as a cream solid.1H NMR (300 MHz, Methanol-d4) δ 9.07 (s, 1H), 8.85 (d, J = 1.4 Hz, 2H), 7.77 (d, J = 1.7 Hz, 1H), 7.48 – 7.27 (m, 2H), 4.70 (s, 2H); LCMS (ESI) m/z: 322.9 [M+H]+. Step 2: Preparation of N-(6-bromo-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]piperidine-1- carboxamide
Figure imgf000265_0001
In a reaction vial, 6-bromo-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine (176 mg, 0.55 mmol) was dissolved in dichloromethane (27 mL) and cooled to 0 °C. To this solution, pyridine (0.27 mL, 3.4 mmol) and triphosgene (81.0 mg, 0.27 mmol) were added and the reaction was stirred at 0 °C for 1 hour. Then, piperidine (51.1 mg, 0.60 mmol) was added at 0 °C and the reaction mixture was warmed to room temperature with stirring overnight. The product was indicated present via UPLC analysis. The reaction mixture was washed with 2x 10 mL deionized water and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product N-(6-bromo- 1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]piperidine-1-carboxamide (146 mg, 0.34 mmol, 62 %) was isolated as a yellow oil.1H NMR (300 MHz, Methanol-d4) δ 9.03 (d, J = 1.1 Hz, 1H), 8.90 (d, J = 1.1 Hz, 2H), 7.93 (t, J = 1.5 Hz, 1H), 7.56 (dd, J = 8.7, 1.1 Hz, 1H), 7.48 (dt, J = 8.7, 1.6 Hz, 1H), 5.18 (s, 2H), 3.45 (t, J = 5.2 Hz, 4H), 1.73 – 1.48 (m, 6H); LCMS (ESI) m/z: 433.0 [M+H]+. Example 219. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]-3- phenoxypropanamide
Figure imgf000265_0002
Step 1: Preparation of 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000265_0003
In a reaction vial, a solution of 1H-imidazole-5-carbaldehyde (113 mg, 1.2 mmol) and 6-fluoro-1,3- benzothiazol-2-amine (200 mg, 1.2 mmol) in dry toluene (12 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry methanol (12 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-20% methanol in ethyl acetate. Product 6-fluoro-N-[(1H-imidazol-5- yl)methyl]-1,3-benzothiazol-2-amine (245 mg, 0.99 mmol, 84 %) was afforded as a pink-brown solid.1H NMR (300 MHz, Methanol-d4) δ 7.66 (d, J = 1.0 Hz, 1H), 7.46 – 7.32 (m, 2H), 7.08 (s, 1H), 7.01 (td, J = 9.1, 2.6 Hz, 1H), 4.57 (s, 2H); LCMS (ESI) m/z: 249.0 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]-3- phenoxypropanamide
Figure imgf000266_0001
In a reaction vial, 6-fluoro-N-[(1H-imidazol-4-yl)methyl]-1,3-benzothiazol-2-amine (217 mg, 0.87 mmol), 3-phenoxypropanoic acid (289 mg, 1.7 mmol), and triethylamine (0.61 mL, 4.4 mmol) were dissolved in dichloromethane (4.5 mL) with stirring. The reaction was cooled to 0 °C, and 50% propylphosphonic anhydride in ethyl acetate (1.7 mL, 2.2 mmol) was added. The reaction was warmed to room temperature and stirred for 3 days. The reaction was diluted with 10 mL deionized water and washed with 2x 10 mL portions of dichloromethane. The organic layers were separated, pooled, and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product N-(6-fluoro-1,3- benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]-3-phenoxypropanamide (244 mg, 0.62 mmol, 71 %) was afforded as a white solid.1H NMR (300 MHz, Methanol-d4) δ 8.28 (s, 1H), 7.86 – 7.69 (m, 1H), 7.64 – 7.53 (m, 1H), 7.34 (s, 1H), 7.31 – 7.06 (m, 4H), 6.95 – 6.78 (m, 3H), 5.59 (s, 2H), 4.51 – 4.25 (m, 2H), 3.39 – 3.22 (m, 2H); LCMS (ESI) m/z: 397.1 [M+H]+. Example 220. Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(4-methylpyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000266_0002
Step 1: Preparation of 6-methoxy-N-[(4-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000266_0003
In a reaction vial, a solution of 4-methylpyridine-3-carbaldehyde (133 mg, 1.1 mmol) and 6- methoxy-1,3-benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (95.7 mg, 2.5 mmol) in dry methanol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-methoxy-N-[(4-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (193 mg, 0.68 mmol, 62 %) was afforded as a white solid.1H NMR (300 MHz, Methanol-d4) δ 8.45 (s, 1H), 8.32 (d, J = 5.1 Hz, 1H), 7.34 (dd, J = 8.8, 1.0 Hz, 1H), 7.29 (d, J = 5.1 Hz, 1H), 7.20 (dd, J = 2.6, 1.0 Hz, 1H), 6.87 (ddd, J = 8.8, 2.7, 1.0 Hz, 1H), 4.66 (s, 2H), 3.79 (d, J = 1.0 Hz, 3H), 2.44 (s, 3H); LCMS (ESI) m/z: 286.0 [M+H]+. Step 2: Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(4-methylpyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000267_0001
In a reaction vial, 6-methoxy-N-[(4-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (194 mg, 0.68 mmol) and N,N-diisopropylethylamine (0.35 mL, 2.0 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (148 mg, 1.0 mmol) was added dropwise with stirring and the reaction was stirred for 16 hours. The product was confirmed present via UPLC analysis. 10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(4-methylpyridin-3- yl)methyl]cyclohexanecarboxamide (90.3 mg, 0.23 mmol, 34 %) was afforded as a white solid. 1H NMR (300 MHz, Methanol-d4) δ 8.31 (d, J = 5.1 Hz, 1H), 7.98 (s, 1H), 7.59 (d, J = 8.9 Hz, 1H), 7.42 (d, J = 2.6 Hz, 1H), 7.36 – 7.26 (m, 1H), 7.00 (d, J = 8.9 Hz, 1H), 5.61 (s, 2H), 3.84 (d, J = 1.1 Hz, 3H), 2.68 (t, J = 11.5 Hz, 1H), 2.51 (s, 3H), 1.86 – 1.48 (m, 7H), 1.36 – 1.12 (m, 3H); LCMS (ESI) m/z: 396.1 [M+H]+. Example 221. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1,3-oxazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000267_0002
Step 1: Preparation of 6-ethyl-N-[(1,3-oxazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000267_0003
In a reaction vial, a solution of 1,3-oxazole-5-carbaldehyde (108 mg, 1.1 mmol) and 6-ethyl-1,3- benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry methanol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-20% methanol in ethyl acetate. Product 6-ethyl-N-[(1,3-oxazol-5- yl)methyl]-1,3-benzothiazol-2-amine (145 mg, 0.56 mmol, 50 %) was afforded as a white solid.1H NMR (300 MHz, Methanol-d4) δ 8.17 (s, 1H), 7.44 (s, 1H), 7.37 (d, J = 8.2 Hz, 1H), 7.12 (d, J = 6.4 Hz, 2H), 4.70 (s, 1H), 2.67 (q, J = 7.6 Hz, 2H), 1.24 (td, J = 7.6, 0.9 Hz, 3H); LCMS (ESI) m/z: 260.0 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1,3-oxazol-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000268_0001
In a reaction vial, 6-ethyl-N-[(1,3-oxazol-5-yl)methyl]-1,3-benzothiazol-2-amine (145 mg, 0.56 mmol) and N,N-diisopropylethylamine (0.29 mL, 1.7 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (244 mg, 1.7 mmol) was added dropwise with stirring and the reaction was stirred for 16 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-50% ethyl acetate in hexanes. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1,3-oxazol-5-yl)methyl]cyclohexanecarboxamide (41.6 mg, 0.11 mmol, 20 %) was afforded as a white solid.1H NMR (300 MHz, Methanol-d4) δ 8.17 (d, J = 1.2 Hz, 1H), 7.72 (d, J = 8.3 Hz, 1H), 7.67 (s, 1H), 7.29 (dd, J = 8.3, 1.8 Hz, 1H), 7.18 (s, 1H), 5.69 (s, 2H), 3.10 (t, J = 11.5 Hz, 1H), 2.76 (q, J = 7.6 Hz, 2H), 1.96 – 1.76 (m, 4H), 1.51 (dq, J = 51.1, 11.6, 11.2 Hz, 6H), 1.34 – 1.21 (m, 3H); LCMS (ESI) m/z: 370.1 [M+H]+. Example 222. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5-yl)methyl]piperidine- 1-carboxamide
Figure imgf000268_0002
Step 1: Preparation of 6-ethyl-N-(thiazol-5-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000268_0003
To a solution of thiazole-5-carbaldehyde (254 mg, 2.2 mmol) and 6-ethylbenzo[d]thiazol-2-amine (400 mg, 2.2 mmol) in toluene (60 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (427 mg, 11 mmol) in methanol (40 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-10% methanol in dichloromethane to offer 6-ethyl-N-(thiazol-5- ylmethyl)benzo[d]thiazol-2-amine (240 mg, 0.87 mmol, 39 %) as a white solid. LCMS (ESI) m/z: 276.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5-yl)methyl]piperidine-1- carboxamide
Figure imgf000269_0001
To a solution of 6-ethyl-N-(thiazol-5-ylmethyl)benzo[d]thiazol-2-amine (100 mg, 0.36 mmol) in tetrahydrofuran (15 mL) at 0 °C was added triethylamine (110 mg, 1.1 mmol) and triphosgene (79.0 mg, 0.72 mmol). The reaction was stirred at 0 °C for 1 hour. Then, piperidine (62.0 mg, 0.73 mmol) was added and the reaction was warmed to room temperature over 16 hours. The product was indicated present via UPLC analysis. The reaction was quenched by ice water (5 mL), then extracted with 3x 15 mL ethyl acetate. The organic layers were pooled, dried over sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by prep-HPLC to give N-(6-ethylbenzo[d]thiazol-2-yl)-N- (thiazol-5-ylmethyl)piperidine-1-carboxamide (9.00 mg, 0.023 mmol, 6.5 %) as a white solid.1H NMR (500 MHz, Chloroform-d) δ 8.68 (s, 1H), 7.89 (s, 1H), 7.67 (d, J = 8.5 Hz, 1H), 7.47 (s, 1H), 7.23 (dd, J = 8.0, 1.5 Hz, 1H), 5.36 (s, 2H), 3.36 (d, J = 5.5 Hz, 4H), 2.72 (q, J = 7.5 Hz, 2H), 1.55-1.59 (m, 6H), 1.27 (t, J = 8.0 Hz, 3H); LCMS (ESI) m/z: 387.1 [M+H]+. Example 223. Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]piperidine-1-carboxamide
Figure imgf000269_0002
Step 1: Preparation of 6-chloro-N-(pyrimidin-5-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000269_0003
To a solution of 6-chlorobenzo[d]thiazol-2-amine (370 mg, 2.0 mmol) and pyrimidine-5- carbaldehyde (260 mg, 2.4 mmol) in toluene (15 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (300 mg, 8.0 mmol) in methanol (10 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-10% methanol in dichloromethane to offer 6-chloro-N- (pyrimidin-5-ylmethyl)benzo[d]thiazol-2-amine (40.0 mg, 0.11 mmol, 14 %) as a white solid. Step 2: Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]piperidine-1- carboxamide
Figure imgf000270_0001
To a solution of triphosgene (17.0 mg, 0.056 mmol) in dichloromethane (3 mL) was added a solution of 6-chloro-N-(pyrimidin-5-ylmethyl)benzo[d]thiazol-2-amine (40.0 mg, 0.14 mmol) and N,N- diisopropylethylamine (54.0 mg, 0.42 mmol) in dichloromethane (3 mL) at room temperature under argon. The mixture was stirred at room temperature for 10 minutes. Then, piperidine (15.0 mg, 0.18 mmol) was added to the reaction vial. The resulting mixture was stirred at room temperature for 4 hours. The product was indicated present via UPLC analysis. Water (5 mL) was added, then the organic layers were extracted with dichloromethane (3x 10 mL). The organic layers were pooled, washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in methanol before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous ammonium bicarbonate) to give the desired product N-(6-chlorobenzo[d]thiazol-2-yl)-N-(pyrimidin-5-ylmethyl)piperidine-1-carboxamide (16.1 mg, 0.040 mmol, 30 %) was afforded as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 9.08 (s, 1H), 8.83 (s, 2H), 8.01 (d, J = 2.1 Hz, 1H), 7.64 (d, J = 8.6 Hz, 1H), 7.39 (dd, J = 8.6, 2.1 Hz, 1H), 5.13 (s, 2H), 3.40 – 3.37 (m, 4H), 1.59 – 1.50 (m, 6H); LCMS (ESI) m/z: 388.0 [M+H]+. Example 224. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-4-(2-methylpropanoyl)-N-[(pyridin-3- yl)methyl]piperazine-1-carboxamide
Figure imgf000270_0002
Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-4-(2-methylpropanoyl)-N-[(pyridin-3-yl)methyl]piperazine- 1-carboxamide
Figure imgf000270_0003
In a reaction vial, a mixture of N-(6-ethylbenzo[d]thiazol-2-yl)-N-(pyridin-3-ylmethyl)piperazine-1- carboxamide (100 mg, 0.26 mmol), isobutyric acid (23.0 mg, 0.26 mmol), N,N-diisopropylethylamine (68.0 mg, 0.52 mmol) and N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (150 mg, 0.39 mmol) in tetrahydrofuran (10 mL) was stirred at 70 °C for 16 hours. The product was indicated present via UPLC analysis. The mixture was poured into water (20 mL) and extracted with 3x 15 mL portions of ethyl acetate. The organic layers were pooled, washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by prep-HPLC to give N-(6-ethylbenzo[d]thiazol-2-yl)-4-isobutyryl-N- (pyridin-3-ylmethyl)piperazine-1-carboxamide (41.0 mg, 0.091 mmol, 35 %) as white solid.1H NMR (500 MHz, Dimethylsulfoxide-d6) δ 8.58 (d, J = 2.0 Hz, 1H), 8.44 (dd, J = 4.75, 1.5 Hz, 1H), 7.76 (d, J = 9.0 Hz, 1H), 7.68 (d, J = 1.0 Hz, 1H), 7.57 (d, J = 8.0 Hz, 1H), 7.33 (q, J = 4.5 Hz, 1H), 7.21 (dd, J = 8.5, 1.5 Hz, 1H), 5.13 (s, 2H), 3.51 (d, J = 2.3 Hz, 4H), 3.38-3.42 (m, 4H), 2.81-2.87 (m, 1H), 2.66 (q, J = 7.5 Hz, 2H), 1.19 (t, J = 7.5 Hz, 3H), 0.98 (d, J = 7.0 Hz, 6H); LCMS (ESI) m/z: 452.3 [M+H]+. Example 225. Preparation of N-(6-cyano-1,3-benzothiazol-2-yl)-3-(3-fluorophenoxy)-N-[(pyridin-3- yl)methyl]propenamide
Figure imgf000271_0001
Step 1: Preparation of 2-(pyridin-3-ylmethylamino)benzo[d]thiazole-6-carbonitrile
Figure imgf000271_0002
To a solution of 2-aminobenzo[d]thiazole-6-carbonitrile (400 mg, 2.3 mmol) and nicotinaldehyde (244 mg, 2.3 mmol) in toluene (16 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (431 mg, 11 mmol) in ethanol (4 mL) was added. The reaction was stirred at 110 °C for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 2-(pyridin-3- ylmethylamino)benzo[d]thiazole-6-carbonitrile as a yellow solid (220 mg, 0.83 mmol, 36 %) as a yellow solid. LCMS data unavailable. Step 2: Preparation of N-(6-cyano-1,3-benzothiazol-2-yl)-3-(3-fluorophenoxy)-N-[(pyridin-3- yl)methyl]propanamide
Figure imgf000271_0003
To a solution of 3-(3-fluorophenoxy)propanoic acid (68.0 mg, 0.37 mmol) in dichloromethane (8 mL) were added 1-hydroxybenzotriazole (74.0 mg, 0.55 mmol) and 1,3-dicyclohexylcarbodiimide (113 mg, 0.55 mmol). The reaction mixture was stirred at room temperature for 2 hours. The resulting solution was cooled to 0 °C, and then a solution of 2-(pyridin-3-ylmethylamino)benzo[d]thiazole-6-carbonitrile (100 mg, 0.37 mmol) in pyridine (43.0 mg, 0.55 mmol) and N,N-dimethylformamide (8 mL) was added. The reaction mixture was stirred at 40 °C for 17 hours. The product was indicated present via UPLC analysis. The solvent was removed under the reduced pressure and the residue was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the desired product N-(6-cyanobenzo[d]thiazol-2-yl)-3-(3-fluorophenoxy)-N-(pyridin-3-ylmethyl)propanamide (22.6 mg, 0.050 mmol, 14 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.62 (dd, J = 6.5, 1.5 Hz, 2H), 8.50 (dd, J = 4.8, 1.4 Hz, 1H), 7.91 (d, J = 8.4 Hz, 1H), 7.83 (dd, J = 8.5, 1.7 Hz, 1H), 7.70 (d, J = 8.2 Hz, 1H), 7.37 (dd, J = 7.9, 4.8 Hz, 1H), 7.30 (dd, J = 15.4, 8.1 Hz, 1H), 6.89 – 6.71 (m, 3H), 5.74 (d, J = 18.5 Hz, 2H), 4.34 (t, J = 5.8 Hz, 2H), 3.30 – 3.16 (m, 2H); LCMS (ESI) m/z: 433.0 [M+H]+. Example 226. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]-1,3- thiazole-2-carboxamide
Figure imgf000272_0001
Step 1: Preparation of 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000272_0002
To a solution of 1H-imidazole-4-carbaldehyde (2.50 g, 27 mmol), 6-fluorobenzo[d]thiazol-2-amine (3.00 g, 18 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (1.30 g, 36 mmol) in methanol (50 mL) was added. The reaction was stirred at 110 °C for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was further purified by silica gel column in a gradient 0-5% methanol in dichloromethane to offer 6-fluoro-N-[(1H-imidazol-5-yl)methyl]-1,3- benzothiazol-2-amine (2.50 g, 10 mmol, 56 %) as a white solid. LCMS (ESI) m/z: 249.1 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]-1,3-thiazole-2- carboxamide
Figure imgf000272_0003
In a reaction vial, a mixture of N-((1H-imidazol-4-yl)methyl)-6-fluorobenzo[d]thiazol-2-amine (150 mg, 0.60 mmol), thiazole-2-carboxylic acid (93.6 mg, 0.72 mmol), N,N-diisopropylethylamine (155 mg, 0.90 mmol) and N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (342 mg, 1.2 mmol) in tetrahydrofuran (10 mL) was stirred at 70 °C for 16 hours. The product was indicated present via UPLC analysis. The mixture was poured into water (20 mL) and extracted with 3x 15 mL portions of ethyl acetate. The organic layers were pooled, washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by prep-HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2 mL/min and the solvent was acetonitrile/0.01% aqueous ammonium carbonate) to afford N-((1H-imidazol- 4-yl)methyl)-N-(6-fluorobenzo[d]thiazol-2-yl)thiazole-2-carboxamide (27.2 mg, 0.076 mmol, 13 %) as white solid.1H NMR (500 MHz, Dimethylsulfoxide-d6) δ 11.85 (s, 1H), 8.22 (d, J = 3.1 Hz, 1H), 8.15 (d, J = 2.9 Hz, 1H), 7.92 (ddd, J = 13.6, 8.7, 3.7 Hz, 2H), 7.43 (s, 1H), 7.34 (td, J = 9.1, 2.6 Hz, 1H), 6.87 (s, 1H), 6.27 (s, 2H); LCMS (ESI) m/z: 360.1 [M+H]+. Example 227. Preparation of tert-butyl 3-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000273_0001
Step 1: Preparation of 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000273_0002
In a round bottom flask equipped with a stir bar, a solution of pyridine-3-carbaldehyde (0.63 mL, 6.7 mmol) and 6-ethyl-1,3-benzothiazol-2-amine (1.20 g, 6.7 mmol) in dry toluene (34 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen overnight. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (1.27 g, 34 mmol) in dry ethyl alcohol (34 mL). The reaction mixture was stirred at 70 °C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with ethyl acetate. The filtrate was concentrated then diluted with dichloromethane and filtered again. The filtrate was purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane through a 40 g column of silica gel. Product 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3- benzothiazol-2-amine (0.915 g, 3.4 mmol, 51 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.72 – 8.50 (m, 2H), 7.76 (dddd, J = 7.9, 2.4, 1.7, 0.8 Hz, 1H), 7.50 – 7.37 (m, 2H), 7.35 – 7.22 (m, 1H), 7.14 (dt, J = 8.3, 1.2 Hz, 1H), 6.24 (s, 1H), 4.68 (s, 2H), 2.70 (q, J = 7.6 Hz, 2H), 1.26 (td, J = 7.6, 0.7 Hz, 3H); LCMS (ESI) m/z: 270.1 [M+H]+. Step 2: Preparation of tert-butyl 3-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000273_0003
In a reaction vial equipped with a stir bar, 6-ethyl-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2- amine (500 mg, 1.9 mmol) was dissolved in dichloromethane (9 mL). To the reaction solution was added 1-[(tert-butoxy)carbonyl]piperidine-3-carboxylic acid (848 mg, 3.7 mmol) and 50% propylphosphonic anhydride in ethyl acetate (3.5 mL, 4.6 mmol). The reaction mixture was cooled to 0 °C and triethylamine (1.3 mL, 9.3 mmol) was added. The reaction was stirred from 0 °C to room temperature overnight. The product was indicated present via UPLC analysis. The reaction solution was diluted with dichloromethane and washed with water (3x 15 mL). The organic layers were pooled, dried over anhydrous sodium sulfate, filtered and concentrated. Crude product was purified via flash column chromatography eluting with 0- 50% ethyl acetate in dichloromethane. Product tert-butyl 3-[(6-ethyl-1,3-benzothiazol-2-yl)[(pyridin-3- yl)methyl]carbamoyl]piperidine-1-carboxylate (176 mg, 0.37 mmol, 20 %) was afforded as a white solid. 1H NMR (300 MHz, Chloroform-d) δ 8.73 – 8.46 (m, 2H), 7.81 – 7.49 (m, 3H), 7.37 – 7.16 (m, 2H), 2.78 (q, J = 7.6 Hz, 5H), 2.02 – 1.63 (m, 3H), 1.60 – 1.07 (m, 16H); LCMS (ESI) m/z: 481.1 [M+H]+. Example 228. Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]piperidine-1-carboxamide
Figure imgf000274_0001
Step 1: Preparation of 6-methoxy-N-(pyrimidin-5-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000274_0002
To a solution of 6-methoxybenzo[d]thiazol-2-amine (400 mg, 2.2 mmol), pyrimidine-5- carbaldehyde (288 mg, 2.7 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (337 mg, 8.8 mmol) in methanol (10 mL) was added. The reaction was stirred at room temperature for 1 hour. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-10% methanol in dichloromethane. Product 6-methoxy-N-(pyrimidin-5-ylmethyl)benzo[d]thiazol-2-amine (245 mg, 0.89 mmol, 40 %) was afforded as a white solid. Step 2: Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]piperidine-1- carboxamide
Figure imgf000274_0003
To a solution of triphosgene (88.0 mg, 0.3 mmol) in dichloromethane (5 mL) was added a solution of piperidine (77.0 mg, 0.9 mmol) and N,N-diisopropylethylamine (116 mg, 0.90 mmol) in dichloromethane (5 mL) at 0 °C under argon. The mixture was stirred at 0 °C for 10 minutes. Then 6-methoxy-N-(pyrimidin- 5-ylmethyl)benzo[d]thiazol-2-amine (90.0 mg, 0.30 mmol) was added at 0 °C. The reaction was warmed to room temperature with stirring over 4 hours. The product was indicated via UPLC analysis. Water (10 mL) was added to the reaction vial, then the reaction was extracted with dichloromethane (3x 20 mL). The organic layers were pooled, washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (the crude samples were dissolved in methanol before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous ammonium bicarbonate) to give the desired product N-(6- methoxybenzo[d]thiazol-2-yl)-N-(pyrimidin-5-ylmethyl)piperidine-1-carboxamide (16.1 mg, 0.042 mmol, 13 %) was afforded as a white solid.1H NMR (400 MHz, Dimethyulsulfoxide-d6) δ 9.07 (s, 1H), 8.81 (s, 2H), 7.57 (d, J = 8.8 Hz, 1H), 7.48 (d, J = 2.6 Hz, 1H), 6.97 (dd, J = 8.8, 2.6 Hz, 1H), 5.10 (s, 2H), 3.77 (s, 3H), 3.37 – 3.32 (m, 4H), 1.57 – 1.49 (m, 6H); LCMS (ESI) m/z: 384.0 [M+H]+. Example 229. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]piperidine- 1-carboxamide
Figure imgf000275_0001
Step 1: Preparation of 6-ethyl-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000275_0002
In a reaction vial, a solution of pyrimidine-5-carbaldehyde (121 mg, 1.1 mmol) and 6-ethyl-1,3- benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry methanol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-ethyl-N-[(pyrimidin-5- yl)methyl]-1,3-benzothiazol-2-amine (163 mg, 0.60 mmol, 54 %) was afforded as a yellow solid.1H NMR (300 MHz, Methanol-d4) δ 9.06 (s, 1H), 8.84 (s, 2H), 7.44 (d, J = 1.9 Hz, 1H), 7.34 (d, J = 8.2 Hz, 1H), 7.11 (dd, J = 8.2, 1.8 Hz, 1H), 4.69 (s, 2H), 2.66 (q, J = 7.6 Hz, 2H), 1.23 (td, J = 7.6, 1.2 Hz, 3H); LCMS (ESI) m/z: 271.0 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]piperidine-1- carboxamide
Figure imgf000275_0003
In a reaction vial, 6-ethyl-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine (163 mg, 0.60 mmol) was dissolved in dichloromethane (30 mL) and cooled to 0 °C. To this solution, pyridine (0.30 mL, 3.7 mmol) and triphosgene (89.3 mg, 0.30 mmol) were added and the reaction was stirred at 0 °C for 1 hour. Then, piperidine (56.3 mg, 0.66 mmol) was added at 0 °C and the reaction mixture was warmed to room temperature with stirring overnight. The product was indicated present via UPLC analysis. The reaction mixture was washed with 2x 10 mL deionized water and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product N-(6-ethyl-1,3- benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]piperidine-1-carboxamide (89.8 mg, 0.24 mmol, 39 %) was isolated as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 9.09 (s, 1H), 8.90 (s, 2H), 7.63 (d, J = 8.3 Hz, 1H), 7.47 (d, J = 1.7 Hz, 1H), 7.22 (dd, J = 8.4, 1.6 Hz, 1H), 5.17 (s, 2H), 3.38 (t, J = 5.1 Hz, 4H), 2.72 (q, J = 7.6 Hz, 2H), 1.74 – 1.46 (m, 6H), 1.26 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 382.1 [M+H]+. Example 230. Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000276_0001
Step 1: Preparation of 6-methoxy-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000276_0002
In a reaction vial, a solution of pyrimidine-5-carbaldehyde (118 mg, 1.1 mmol) and 6-methoxy-1,3- benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (95.7 mg, 2.5 mmol) in dry methanol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-methoxy-N-[(pyrimidin- 5-yl)methyl]-1,3-benzothiazol-2-amine (172 mg, 0.63 mmol, 58 %) was afforded as a pink solid.1H NMR (300 MHz, Methanol-d4) δ 9.06 (s, 1H), 8.84 (s, 2H), 7.33 (dd, J = 8.8, 1.1 Hz, 1H), 7.21 (dd, J = 2.8, 1.1 Hz, 1H), 6.87 (ddd, J = 8.8, 2.7, 1.2 Hz, 1H), 4.67 (s, 2H), 3.79 (d, J = 1.2 Hz, 3H); LCMS (ESI) m/z: 273.0 [M+H]+. Step 2: Preparation of N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000276_0003
In a reaction vial, 6-methoxy-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine (172 mg, 0.63 mmol) and N,N-diisopropylethylamine (0.33 mL, 1.9 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (277 mg, 1.9 mmol) was added dropwise with stirring and the reaction was stirred for 16 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-45% ethyl acetate in hexanes. Product N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]cyclohexanecarboxamide (191 mg, 0.50 mmol, 79 %) was afforded as a white solid. 1H NMR (300 MHz, Chloroform-d) δ 9.16 (d, J = 2.6 Hz, 1H), 8.74 (t, J = 2.0 Hz, 2H), 7.81 – 7.59 (m, 1H), 7.32 – 7.20 (m, 1H), 7.02 (d, J = 8.9 Hz, 1H), 5.52 (s, 2H), 3.86 (q, J = 2.1 Hz, 3H), 2.69 (d, J = 12.0 Hz, 1H), 1.80 (d, J = 13.0 Hz, 5H), 1.69 (s, 5H); LCMS (ESI) m/z: 383.1 [M+H]+. Example 231. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000277_0001
Step 1: Preparation of 6-fluoro-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine In a reaction vial, a solution of pyrimidine-5-carbaldehyde (127 mg, 1.2 mmol) and 6-fluoro-1,3- benzothiazol-2-amine (200 mg, 1.2 mmol) in dry toluene (12 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry methanol (12 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-fluoro-N-[(pyrimidin-5- yl)methyl]-1,3-benzothiazol-2-amine (149 mg, 0.57 mmol, 49 %) was afforded as a yellow solid.1H NMR (300 MHz, Methanol-d4) δ 9.07 (d, J = 1.3 Hz, 1H), 8.85 (d, J = 1.3 Hz, 2H), 7.63 – 7.27 (m, 2H), 7.02 (tdd, 0.1J = 9.1, 2.8, 1.4 Hz, 1H), 4.70 (s, 2H); LCMS (ESI) m/z: 261.0 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]cyclohexanecarboxamide
Figure imgf000277_0002
In a reaction vial, 6-fluoro-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine (149 mg, 0.57 mmol) and N,N-diisopropylethylamine (0.30 mL, 1.7 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (250 mg, 1.7 mmol) was added dropwise with stirring and the reaction was stirred for 16 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-60% ethyl acetate in hexanes. Product N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]cyclohexanecarboxamide (162 mg, 0.44 mmol, 77 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 9.18 (d, J = 4.8 Hz, 1H), 8.75 (d, J = 4.8 Hz, 2H), 7.70 (dt, J = 9.2, 4.7 Hz, 1H), 7.50 (ddd, J = 7.9, 4.9, 2.5 Hz, 1H), 7.16 (ddt, J = 11.3, 6.1, 2.5 Hz, 1H), 5.56 (s, 2H), 2.69 (d, J = 12.1 Hz, 1H), 1.96 – 1.72 (m, 5H), 1.77 – 1.58 (m, 4H), 1.39 – 1.26 (m, 1H); LCMS (ESI) m/z: 371.0 [M+H]+. Example 232. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]piperidine-1-carboxamide
Figure imgf000278_0001
Step 1: Preparation of 6-fluoro-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000278_0002
In a reaction vial, a solution of pyrimidine-5-carbaldehyde (127 mg, 1.2 mmol) and 6-fluoro-1,3- benzothiazol-2-amine (200 mg, 1.2 mmol) in dry toluene (12 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry methanol (12 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-fluoro-N-[(pyrimidin-5- yl)methyl]-1,3-benzothiazol-2-amine (149 mg, 0.57 mmol, 49 %) was afforded as a yellow solid.1H NMR (300 MHz, Methanol-d4) δ 9.07 (d, J = 1.3 Hz, 1H), 8.85 (d, J = 1.3 Hz, 2H), 7.63 – 7.27 (m, 2H), 7.02 (tdd, 0.1J = 9.1, 2.8, 1.4 Hz, 1H), 4.70 (s, 2H); LCMS (ESI) m/z: 261.0 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]piperidine-1- carboxamide
Figure imgf000278_0003
In a reaction vial, 6-fluoro-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine (171 mg, 0.65 mmol) was dissolved in dichloromethane (33 mL) and cooled to 0 °C. To this solution, pyridine (0.32 mL, 4.0 mmol) and triphosgene (96.7 mg, 0.33 mmol) were added and the reaction was stirred at 0 °C for 1 hour. Then, piperidine (61.1 mg, 0.72 mmol) was added at 0 °C and the reaction mixture was warmed to room temperature with stirring overnight. The product was indicated present via UPLC analysis. The reaction mixture was washed with 2x 10 mL deionized water and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-70% ethyl acetate in hexanes. Product N-(6-fluoro-1,3- benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]piperidine-1-carboxamide (162 mg, 0.44 mmol, 67 %) was isolated as a white solid.1H NMR (300 MHz, Chloroform-d) δ 9.11 (d, J = 2.0 Hz, 1H), 8.90 (d, J = 2.0 Hz, 2H), 7.70 – 7.53 (m, 1H), 7.35 (dd, J = 8.1, 2.4 Hz, 1H), 7.15 – 7.02 (m, 1H), 5.17 (d, J = 2.0 Hz, 2H), 3.39 (d, J = 5.6 Hz, 4H), 1.60 (s, 6H); LCMS (ESI) m/z: 372.1 [M+H]+. Example 233. Preparation of 4-cyano-N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]benzamide
Figure imgf000279_0001
Step 1: Preparation of 6-methoxy-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000279_0002
In a reaction vial equipped with a magnetic stir bar, a solution of pyrimidine-5-carbaldehyde (141 mg, 1.3 mmol) and 6-methoxy-1,3-benzothiazol-2-amine (250 mg, 1.4 mmol) in dry toluene (7 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen for 24 hours. The reaction mixture was cooled to 60 °C and then to the reaction was added sodium borohydride (119 mg, 3.2 mmol) and dry methanol (7 mL). The reaction mixture was stirred at 70 °C for 1 hour and then cooled to room temperature. The reaction was filtered over a bed of Celite and washed with methanol. The filtrate was concentrated then diluted with dichloromethane and re-filtered. The filtrate was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane. Product 6-methoxy-N-[(pyrimidin-5- yl)methyl]-1,3-benzothiazol-2-amine (155 mg, 0.57 mmol, 41 %) was isolated as an orange solid.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 9.10 (d, J = 1.0 Hz, 1H), 8.83 (d, J = 1.0 Hz, 1H), 8.36 (t, J = 5.7 Hz, 1H), 7.41 – 7.16 (m, 1H), 6.83 (ddd, J = 8.8, 2.7, 1.1 Hz, 1H), 4.59 (d, J = 5.7 Hz, 2H), 3.72 (q, J = 2.2, 1.6 Hz, 2H), 3.40 – 3.18 (m, 1H); LCMS (ESI) m/z: 272.9 [M+H]+. Step 2: Preparation of 4-cyano-N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]benzamide
Figure imgf000279_0003
In a reaction vial equipped with a magnetic stir bar, 6-methoxy-N-[(pyrimidin-5-yl)methyl]-1,3- benzothiazol-2-amine (156 mg, 0.57 mmol) was dissolved in tetrahydrofuran (3 mL) and cooled to 0 °C. To the solution was added triethylamine (0.20 mL, 1.4 mmol) then 4-cyanobenzoyl chloride (141 mg, 0.85 mmol) was added, slowly. The reaction was stirred from 0 °C to room temperature overnight. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane and washed with water (3x). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Crude product was purified via flash column chromatography eluting with 0-100% ethyl acetate in hexanes. Product 4-cyano-N-(6-methoxy-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]benzamide (87.9 mg, 0.22 mmol, 39 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 9.16 (s, 1H), 8.72 (s, 2H), 7.74 (dd, J = 8.6, 4.2 Hz, 3H), 7.67 – 7.51 (m, 2H), 7.35 – 7.19 (m, 1H), 7.08 (dd, J = 8.9, 2.6 Hz, 1H), 5.42 (s, 2H), 3.88 (s, 3H); LCMS (ESI) m/z: 402.1 [M+H]+. Example 234. Preparation of tert-butyl 4-[(6-methoxy-1,3-benzothiazol-2-yl)[(pyridin-3- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000280_0001
Step 1: Preparation of 6-methoxy-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000280_0002
In a reaction vial, a solution of pyridine-3-carbaldehyde (117 mg, 1.1 mmol) and 6-methoxy-1,3- benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (95.7 mg, 2.5 mmol) in dry methanol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-methoxy-N-[(pyridin-3- yl)methyl]-1,3-benzothiazol-2-amine (248 mg, 0.92 mmol, 83 %) was afforded as a yellow solid.1H NMR (300 MHz, Methanol-d4) δ 8.70 – 8.46 (m, 1H), 8.54 – 8.37 (m, 1H), 7.98 – 7.79 (m, 1H), 7.43 (dd, J = 7.9, 5.0 Hz, 1H), 7.33 (dd, J = 8.9, 1.1 Hz, 1H), 7.20 (dd, J = 2.7, 1.1 Hz, 1H), 6.87 (ddd, J = 8.8, 2.7, 1.2 Hz, 1H), 4.67 (s, 2H), 3.79 (d, J = 1.3 Hz, 3H); LCMS (ESI) m/z: 272.0 [M+H]+. Step 2: Preparation of tert-butyl 4-[(6-methoxy-1,3-benzothiazol-2-yl)[(pyridin-3- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000280_0003
In a reaction vial, 6-methoxy-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (249 mg, 0.92 mmol), 1-[(tert-butoxy)carbonyl]piperidine-4-carboxylic acid (419 mg, 1.8 mmol), and triethylamine (0.64 mL, 4.6 mmol) were dissolved in dichloromethane (5 mL) with stirring. The reaction was cooled to 0 °C, and 50% propylphosphonic anhydride in ethyl acetate (1.7 mL, 2.3 mmol) was added. The reaction was warmed to room temperature and stirred for 3 days. The product was indicated via UPLC analysis. The reaction was diluted with 30 mL deionized water and 30 mL dichloromethane. The organic layer was separated, washed with 2 x 10 mL portions deionzed water, pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-40% ethyl acetate in hexanes. Product tert-butyl 4-[(6-methoxy-1,3-benzothiazol-2- yl)[(pyridin-3-yl)methyl]carbamoyl]piperidine-1-carboxylate (78.0 mg, 0.16 mmol, 18 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.61 (s, 1H), 8.56 (dd, J = 4.9, 1.4 Hz, 1H), 7.76 – 7.55 (m, 2H), 7.29 (dd, J = 7.2, 3.8 Hz, 2H), 7.04 (d, J = 9.0 Hz, 1H), 4.12 (s, 2H), 3.87 (s, 3H), 2.84 (s, 1H), 2.67 (s, 2H), 1.75 (ddd, J = 33.9, 18.5, 9.0 Hz, 5H), 1.45 (s, 9H); LCMS (ESI) m/z: 427.1 [M-t-butyl]+. Example 235. Preparation of 4-fluoro-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]benzamide
Figure imgf000281_0001
Step 1: Preparation of 6-fluoro-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000281_0002
In a reaction vial equipped with a magnetic stir bar, a solution of pyrimidine-5-carbaldehyde (151 mg, 1.4 mmol) and 6-fluoro-1,3-benzothiazol-2-amine (250 mg, 1.5 mmol) in dry toluene (7 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen for 24 hours. The reaction mixture was cooled to 60 °C and then to the reaction was added sodium borohydride (128 mg, 3.4 mmol) and dry methanol (7 mL). The reaction mixture was stirred at 70 °C for 1 hour and then cooled to room temperature. The reaction was filtered over a bed of Celite and washed with methanol. Filtrate was concentrated then diluted with dichloromethane and re-filtered. The filtrate was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane. Product 6-fluoro-N-[(pyrimidin-5- yl)methyl]-1,3-benzothiazol-2-amine (154 mg, 0.59 mmol, 40 %) was afforded as an olive green solid.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 9.10 (s, 1H), 8.83 (s, 2H), 8.57 (t, J = 5.7 Hz, 1H), 7.63 (dd, J = 8.7, 2.7 Hz, 1H), 7.39 (dd, J = 8.8, 4.8 Hz, 1H), 7.14 – 6.93 (m, 1H), 4.62 (d, J = 5.6 Hz, 2H), 3.39 – 3.21 (m, 1H); LCMS (ESI) m/z: 261.0 [M+H]+. Step 2: Preparation of 4-fluoro-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]benzamide
Figure imgf000282_0001
In a reaction vial equipped with a magnetic stir bar, 6-fluoro-N-[(pyrimidin-5-yl)methyl]-1,3- benzothiazol-2-amine (155 mg, 0.59 mmol) was dissolved in tetrahydrofuran (3 mL) and cooled to 0 °C. To the solution was added triethylamine (0.20 mL, 1.5 mmol) then 4-fluorobenzoyl chloride (140 mg, 0.89 mmol) was added, slowly. The reaction was stirred from 0 °C to room temperature for 2.5 hours. The product was indicated present via UPLC analysis. The reaction was diluted with dichloromethane and washed with water (3x). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Crude product was purified via flash column chromatography eluting with 0-100% ethyl acetate in hexanes. Product 4-fluoro-N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]benzamide (181 mg, 0.47 mmol, 80 %) was afforded as a beige solid.1H NMR (300 MHz, Chloroform-d) δ 9.14 (s, 1H), 8.66 (s, 2H), 7.78 (dd, J = 8.9, 4.7 Hz, 1H), 7.62 – 7.43 (m, 3H), 7.32 – 7.09 (m, 3H), 5.50 (s, 2H); LCMS (ESI) m/z: 383.0 [M+H]+. Example 236. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(4-methylpyridin-3- yl)methyl]piperidine-1-carboxamide
Figure imgf000282_0002
Step 1: Preparation of 6-ethyl-N-((4-methylpyridin-3-yl)methyl)benzo[d]thiazol-2-amine
Figure imgf000282_0003
To a solution of 6-ethylbenzo[d]thiazol-2-amine (200 mg, 1.1 mmol) and 4-methylnicotinaldehyde (136 mg, 1.1 mmol) in toluene (50 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (213 mg, 5.6 mmol) in methanol (30 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-10% methanol in dichloromethane. Product 6-ethyl-N-((4-methylpyridin-3-yl)methyl)benzo[d]thiazol-2-amine as white solid (140 mg, 0.49 mmol, 45 %); LCMS (ESI) m/z: 284.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(4-methylpyridin-3-yl)methyl]piperidine-1- carboxamide
Figure imgf000283_0001
To a solution of 6-ethyl-N-((4-methylpyridin-3-yl)methyl)benzo[d]thiazol-2-amine (120 mg, 0.42 mmol) in tetrahydrofuran (15 mL) was added triethylamine (128 mg, 1.3 mmol) and triphosgene (250 mg, 0.85 mmol). The mixture was stirred at 0 °C for 1 hour. Then, piperidine (72.0 mg, 0.85 mmol) was added, and the reaction was warmed to room temperature with stirring over 16 hours. The product was indicated present via UPLC analysis. The reaction was quenched by ice water (20 mL), then extracted with 3x 30 mL portions of ethyl acetate. The organic layers were pooled, dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC to give N-(6- ethylbenzo[d]thiazol-2-yl)-N-((4-methylpyridin-3-yl)methyl)piperidine-1-carboxamide (8.80 mg, 0.022 mmol, 5.3 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.41 (s, 1H), 8.32 (d, J = 4.8 Hz, 1H), 7.68 (d, J = 1.2 Hz, 1H), 7.54 (d, J = 8.0 Hz, 1H), 7.20 (t, J = 6.0 Hz, 2H), 5.12 (s, 2H), 3.33 (d, J = 5.6 Hz, 4H), 2.66 (q, J = 7.6 Hz, 2H), 2.38 (s, 3H), 1.53 (d, J = 4.4 Hz, 2H), 1.41 (d, J = 3.6 Hz, 4H), 1.19 (t, J = 7.6 Hz, 3H); LCMS (ESI) m/z: 395.3 [M+H]+. Example 237. Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(6-methylpyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000283_0002
Step 1: Preparation of 6-ethyl-N-[(6-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000283_0003
In a reaction vial, a solution of 6-methylpyridine-3-carbaldehyde (135 mg, 1.1 mmol) and 6-ethyl- 1,3-benzothiazol-2-amine (200 mg, 1.1 mmol) in dry toluene (11 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (97.2 mg, 2.6 mmol) in dry ethyl alcohol (11 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-ethyl-N-[(6-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (247 mg, 0.87 mmol, 78 %) was afforded as a white solid.1H NMR (300 MHz, Methanol-d4) δ 8.44 (d, J = 2.0 Hz, 1H), 7.77 (dd, J = 8.0, 2.2 Hz, 1H), 7.43 (s, 1H), 7.32 (dd, J = 15.2, 8.1 Hz, 2H), 7.11 (d, J = 8.3 Hz, 1H), 4.63 (s, 2H), 2.67 (q, J = 7.5 Hz, 2H), 2.51 (s, 3H), 1.24 (td, J = 7.6, 1.3 Hz, 3H); LCMS (ESI) m/z: 284.1 [M+H]+. Step 2: Preparation of N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(6-methylpyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000284_0001
In a reaction vial, 6-ethyl-N-[(6-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (248 mg, 0.87 mmol) and N,N-diisopropylethylamine (0.45 mL, 2.6 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. cyclohexanecarbonyl chloride (382 mg, 2.6 mmol) was added dropwise with stirring and the reaction was stirred for 16 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-(6-ethyl-1,3-benzothiazol-2-yl)-N-[(6-methylpyridin-3- yl)methyl]cyclohexanecarboxamide (202 mg, 0.51 mmol, 59 %) was afforded as a yellow solid. 1H NMR (300 MHz, Chloroform-d) δ 8.49 (d, J = 2.4 Hz, 1H), 7.68 (d, J = 8.3 Hz, 1H), 7.62 (d, J = 1.7 Hz, 1H), 7.46 (dd, J = 8.2, 1.9 Hz, 1H), 7.25 (dd, J = 8.3, 1.7 Hz, 1H), 7.08 (d, J = 8.1 Hz, 1H), 5.56 (s, 2H), 2.76 (q, J = 7.6 Hz, 2H), 2.74 – 2.62 (m, 1H), 2.54 (s, 3H), 1.71 (dt, J = 39.2, 11.1 Hz, 7H), 1.29 (t, J = 7.6 Hz, 3H), 1.27 – 1.14 (m, 3H); LCMS (ESI) m/z: 394.2 [M+H]+. Example 238. Preparation of N-[6-(2-aminoethoxy)-1,3-benzothiazol-2-yl]-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000284_0002
Preparation of N-[6-(2-aminoethoxy)-1,3-benzothiazol-2-yl]-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000284_0003
In a reaction vial, tert-butyl (2-((2-(N-(pyridin-3- ylmethyl)cyclohexanecarboxamido)benzo[d]thiazol-6-yl)oxy)ethyl)carbamate (65.0 mg, 0.13 mmol) was dissolved in dichloromethane (5 mL). Trifluoroacetic acid (2 mL) was added and the reaction was stirred at room temperature for 1 hour. The product was indicated present via UPLC analysis. The reaction was concentrated under reduced pressure, and the residue was quenched with a 7 M solution of ammonia in methanol until pH 7. The mixture was concentrated under reduced pressure and crude product was purified by prep-HPLC (SunFire C18, 4.6*50mm, 3.5um column Xbridge C183.5μm 4.6×50mm column. The elution system used was a gradient of 5%-95% over 1.5 min at 2ml/min and the solvent was acetonitrile/0.01% aqueous ammonia bicarbonate). Product N-(6-(2-aminoethoxy)benzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)cyclohexanecarboxamide (30.0 mg, 0.073 mmol, 56 %) was afforded as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.56 (s, 1H), 8.51 – 8.46 (m, 1H), 7.69 – 7.53 (m, 3H), 7.37 (dd, J = 7.5, 4.8 Hz, 1H), 7.02 (dd, J = 8.9, 2.5 Hz, 1H), 5.65 (s, 2H), 4.03 – 3.88 (m, 2H), 2.95 – 2.75 (m, 3H), 1.64 (d, J = 11.6 Hz, 5H), 1.52 – 1.33 (m, 2H), 1.21 (t, J = 10.9 Hz, 3H); LCMS (ESI) m/z: 411.1 [M+H]+. Example 239. Preparation of 1-cyclopropanecarbonyl-N-[6-(cyclopropylmethoxy)-1,3-benzothiazol- 2-yl]-N-[(pyridin-3-yl)methyl]piperidine-4-carboxamide
Figure imgf000285_0001
Step 1: Preparation of 6-(cyclopropylmethoxy)-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000285_0002
To a solution of 6-(cyclopropylmethoxy)benzo[d]thiazol-2-amine (180 mg, 0.82 mmol) and nicotinaldehyde (131 mg, 1.2 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (426 mg, 11 mmol) in methanol (30 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-20% methanol in dichloromethane. Product 6-(cyclopropylmethoxy)-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (127 mg, 0.41 mmol, 50 %) as a white solid. LCMS (ESI) m/z: 312.1 [M+H]+. Step 2: Preparation of 1-(cyclopropanecarbonyl)-N-(6-(cyclopropylmethoxy)benzo[d]thiazol-2-yl)-N- (pyridin-3-ylmethyl)piperidine-4-carboxamide
Figure imgf000285_0003
To a solution of 6-(cyclopropylmethoxy)-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2-amine (127 mg, 0.41 mmol) in tetrahydrofuran (5 mL) were added 1-(cyclopropanecarbonyl)piperidine-4-carboxylic acid (80.0 mg, 0.41 mmol) and N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (233 mg, 0.61 mmol) and N,N- diisopropylethylamine (105 mg, 0.82 mmol). The reaction was stirred at 70 °C for 16 hours. The product was indicated via UPLC analysis. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the product 1-cyclopropanecarbonyl-N- [6-(cyclopropylmethoxy)-1,3-benzothiazol-2-yl]-N-[(pyridin-3-yl)methyl]piperidine-4-carboxamide (28.6 mg, 0.068 mmol, 14 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.58 (d, J = 1.2 Hz, 1H), 8.49 (dd, J = 4.0 Hz, 1H), 7.61-7.65 (m, 2H), 7.54 (d, J = 2.0Hz, 1H), 7.36-7.38 (m, 1H), 7.00-7.02 (m, 1H), 5.68 (s, 2H), 4.25-4.36 (m, 2H), 3.85-3.86 (m, 2H), 3.10-3.26 (m, 2H), 2.61-2.64 (m, 1H), 1.94-1.99 (m, 1H), 1.45-1.76 (m, 4H), 1.21-1.27 (m, 1H), 0.70-0.71 (m, 4H), 0.56-0.59 (m, 2H), 0.31-0.34 (m, 2H); LCMS (ESI) m/z: 491.3 [M+H]+. Example 240. Preparation of methyl 4-[(6-cyano-1,3-benzothiazol-2-yl)[(1H-imidazol-4- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000286_0001
Step 1: Preparation of 2-({[1-(triphenylmethyl)-1H-imidazol-4-yl]methyl}amino)-1,3-benzothiazole-6- carbonitrile
Figure imgf000286_0002
In a reaction vial equipped with a magnetic stir bar, a solution of 1-(triphenylmethyl)-1H- imidazole-4-carbaldehyde (453 mg, 1.3 mmol) and 2-amino-1,3-benzothiazole-6-carbonitrile (250 mg, 1.4 mmol) in dry toluene (7 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen for 24 hours. The reaction mixture was cooled to 60 °C and then to the reaction was added sodium borohydride (123 mg, 3.3 mmol) and dry methanol (7 mL). The reaction mixture was stirred at 64°C for 1 hour and then cooled to room temperature. The reaction was filtered over a bed of Celite and washed with methanol. The filtrate was concentrated then diluted with dichloromethane and re-filtered. The filtrate was purified via flash column chromatography eluting with 0-100% ethyl acetate in hexanes. Product 2- ({[1-(triphenylmethyl)-1H-imidazol-4-yl]methyl}amino)-1,3-benzothiazole-6-carbonitrile (124 mg, 0.25 mmol, 18 %) was isolated as yellow solid.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 8.80 (s, 1H), 8.19 (t, J = 1.6 Hz, 1H), 7.62 (dt, J = 8.4, 1.7 Hz, 1H), 7.54 – 7.27 (m, 12H), 7.22 – 7.01 (m, 6H), 6.91 (d, J = 1.6 Hz, 1H), 4.46 (d, J = 5.2 Hz, 2H). Step 2: Preparation of methyl 4-[(6-cyano-1,3-benzothiazol-2-yl)({[1-(triphenylmethyl)-1H-imidazol-4- yl]methyl})carbamoyl]piperidine-1-carboxylate
Figure imgf000287_0001
In a reaction vial equipped with a stir bar, 2-({[1-(triphenylmethyl)-1H-imidazol-4-yl]methyl}amino)- 1,3-benzothiazole-6-carbonitrile (124 mg, 0.25 mmol) was dissolved in dichloromethane (1.5 mL). To the reaction solution was added 1-(methoxycarbonyl)piperidine-4-carboxylic acid (93.2 mg, 0.50 mmol) and 50% propylphosphonic anhydride in ethyl acetate (0.47 mL, 0.62 mmol). The reaction mixture was cooled to 0 °C and triethylamine (0.17 mL, 1.2 mmol) was added. The reaction was stirred from 0 °C to room temperature overnight. The product was indicated present via UPLC analysis. The reaction solution was diluted with dichloromethane and washed with water (3 x 15 mL). The organic layers were pooled, dried over anhydrous sodium sulfate, filtered and concentrated. Crude product was purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane. Product methyl 4-[(6-cyano-1,3- benzothiazol-2-yl)({[1-(triphenylmethyl)-1H-imidazol-4-yl]methyl})carbamoyl]piperidine-1-carboxylate (150 mg, 0.23 mmol, 90 %) was afforded as a white solid. Step 3: Preparation of methyl 4-[(6-cyano-1,3-benzothiazol-2-yl)[(1H-imidazol-4- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000287_0002
In a reaction vial equipped with a stir bar, methyl 4-[(6-cyano-1,3-benzothiazol-2-yl)({[1- (triphenylmethyl)-1H-imidazol-4-yl]methyl})carbamoyl]piperidine-1-carboxylate (151 mg, 0.22 mmol) was dissolved in dichloromethane (1.5 mL). To the reaction was added trifluoroacetic acid (0.14 mL, 1.8 mmol) and the reaction was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The reaction was concentrated under reduced pressure then diluted with dichloromethane and quenched with saturated sodium bicarbonate until bubbling ceased. The reaction was diluted with 10 mL deionized water and washed with dichloromethane (2x 10 mL). The organic layers were combined and dried over anhydrous sodium sulfate, filtered, then concentrated. Crude product was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane. Product methyl 4-[(6-cyano-1,3-benzothiazol-2-yl)[(1H-imidazol-4-yl)methyl]carbamoyl]piperidine-1-carboxylate (44.4 mg, 0.10 mmol, 44 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.15 (dd, J = 1.6, 0.7 Hz, 1H), 7.90 (dd, J = 8.4, 0.7 Hz, 1H), 7.76 – 7.58 (m, 2H), 7.27 (d, J = 0.8 Hz, 1H), 7.16 (s, 1H), 5.57 (s, 2H), 4.25 (s, 2H), 3.73 (d, J = 0.8 Hz, 4H), 2.99 (s, 2H), 1.85 (d, J = 3.8 Hz, 4H); LCMS (ESI) m/z: 425.1 [M+2H]+. Example 241. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyrimidin-5- yl)methyl]propenamide
Figure imgf000288_0001
Step 1: Preparation of 5-fluoro-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000288_0002
In a reaction vial, a solution of pyrimidine-5-carbaldehyde (192 mg, 1.8 mmol) and 5-fluoro-1,3- benzothiazol-2-amine (300 mg, 1.8 mmol) in dry toluene (18 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (154 mg, 4.1 mmol) in dry ethyl alcohol (18 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12- gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 5-fluoro-N- [(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine (193 mg, 0.74 mmol, 42 %) was afforded as an orange solid.1H NMR (300 MHz, Methanol-d4) δ 9.07 (d, J = 1.4 Hz, 1H), 8.85 (d, J = 1.3 Hz, 2H), 7.56 (ddd, J = 8.8, 5.4, 1.3 Hz, 1H), 7.14 (ddd, J = 10.1, 2.6, 1.3 Hz, 1H), 7.00 – 6.76 (m, 1H), 4.71 (s, 2H); LCMS (ESI) m/z: 261.0 [M+H]+. Step 2: Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(pyrimidin-5- yl)methyl]propanamide
Figure imgf000288_0003
In a reaction vial, 5-fluoro-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine (95.9 mg, 0.37 mmol), 3-phenoxypropanoic acid (122 mg, 0.74 mmol), and triethylamine (0.26 mL, 1.8 mmol) were dissolved in dichloromethane (2 mL) with stirring. The reaction was cooled to 0 °C, and 50% propylphosphonic anhydride in ethyl acetate (0.70 mL, 0.92 mmol) was added. The reaction was warmed to room temperature and stirred for 2.5 days. The reaction was diluted with 10 mL deionized water and washed with 2x 10 mL portions of dichloromethane. The organic layers were separated, pooled, and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product N-(5-fluoro-1,3- benzothiazol-2-yl)-3-phenoxy-N-[(pyrimidin-5-yl)methyl]propanamide (54.4 mg, 0.13 mmol, 36 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 9.19 (s, 1H), 8.80 (s, 2H), 7.75 (dd, J = 8.7, 5.2 Hz, 1H), 7.47 (dd, J = 9.6, 2.5 Hz, 1H), 7.35 – 7.23 (m, 2H), 7.11 (td, J = 8.9, 2.5 Hz, 1H), 7.02 – 6.94 (m, 1H), 6.94 – 6.82 (m, 2H), 5.65 (s, 2H), 4.43 (t, J = 6.2 Hz, 2H), 3.15 (t, J = 6.2 Hz, 2H); LCMS (ESI) m/z: 409.0 [M+H]+. Example 242. Preparation of 1-cyclopropanecarbonyl-N-[6-(cyclopropylmethoxy)-1,3-benzothiazol- 2-yl]-N-[(pyrimidin-5-yl)methyl]piperidine-4-carboxamide
Figure imgf000289_0001
Step 1: Preparation of 6-(cyclopropylmethoxy)-N-(pyrimidin-5-ylmethyl)benzo[d]thiazol-2-amine
Figure imgf000289_0002
To a solution of 6-(cyclopropylmethoxy)benzo[d]thiazol-2-amine (190 mg, 0.86 mmol) and pyrimidine-5-carbaldehyde (140 mg, 1.3 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (426 mg, 11 mmol) in methanol (30 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-20% methanol in dichloromethane. Product 6-(cyclopropylmethoxy)-N-(pyrimidin-5-ylmethyl)benzo[d]thiazol-2- amine (92.0 mg, 0.30 mmol, 33 %) as a white solid. LCMS (ESI) m/z: 313.2 [M+H]+. Step 2: Preparation of 1-cyclopropanecarbonyl-N-[6-(cyclopropylmethoxy)-1,3-benzothiazol-2-yl]-N- [(pyrimidin-5-yl)methyl]piperidine-4-carboxamide
Figure imgf000289_0003
To a solution of 6-(cyclopropylmethoxy)-N-(pyrimidin-5-ylmethyl)benzo[d]thiazol-2-amine (92.0 mg, 0.30 mmol) in tetrahydrofuran (5 mL) were added 1-(cyclopropanecarbonyl)piperidine-4-carboxylic acid (58.0 mg, 0.29 mmol) and N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N- methylmethanaminium hexafluorophosphate N-oxide (168 mg, 0.44 mmol) and N,N- diisopropylethylamine (76.0 mg, 0.58 mmol). The reaction was stirred at 70 °C for 16 hours. The product was indicated via UPLC analysis. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the product 1-cyclopropanecarbonyl-N- [6-(cyclopropylmethoxy)-1,3-benzothiazol-2-yl]-N-[(pyrimidin-5-yl)methyl]piperidine-4-carboxamide (4.9 mg, 0.010 mmol, 3.0 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 9.12 (s, 1H), 8.80 (s, 2H), 7.54-7.63 (m, 2H), 7.01 (dd, J = 6.8Hz, 1H), 5.64 (s, 2H), 4.27-4.38 (m, 2H), 3.85 (d, J = 2.0 Hz, 2H), 3.28-3.29 (m, 1H), 3.16-3.18 (m, 1H), 2.64-2.69 (m, 1H), 1.97-2.01 (m, 1H), 1.78-1.86 (m, 2H), 1.47-1.63 (m, 2H), 1.21-1.26 (m, 1H), 0.71-0.72 (m, 4H), 0.56-0.59 (m, 2H), 0.31-0.34 (m, 2H); LCMS (ESI) m/z: 492.2 [M+H]+. Example 243. Preparation of 1-cyclopropanecarbonyl-N-[6-(cyclopropylmethoxy)-1,3-benzothiazol- 2-yl]-N-[(1H-imidazol-5-yl)methyl]piperidine-4-carboxamide
Figure imgf000290_0001
Step 1: Preparation of N-((1H-imidazol-5-yl)methyl)-6-(cyclopropylmethoxy)benzo[d]thiazol-2-amine
Figure imgf000290_0002
To a solution of 6-(cyclopropylmethoxy)benzo[d]thiazol-2-amine (180 mg, 0.82 mmol) and 1H- imidazole-5-carbaldehyde (118 mg, 1.2 mmol) in toluene (20 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (426 mg, 11 mmol) in methanol (30 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-20% methanol in dichloromethane. Product 6-(cyclopropylmethoxy)-N-(pyridin-3-ylmethyl)benzo[d]thiazol-2- amine (38.8 mg, 0.13 mmol, 16 %) as a white solid. LCMS (ESI) m/z: 301.0 [M+H]+. Step 2: Preparation of 1-cyclopropanecarbonyl-N-[6-(cyclopropylmethoxy)-1,3-benzothiazol-2-yl]-N-[(1H- imidazol-5-yl)methyl]piperidine-4-carboxamide
Figure imgf000290_0003
To a solution of N-((1H-imidazol-5-yl)methyl)-6-(cyclopropylmethoxy)benzo[d]thiazol-2-amine (38.8 mg, 0.13 mmol) in tetrahydrofuran (5 mL) were added 1-(cyclopropanecarbonyl)piperidine-4- carboxylic acid (25.0 mg, 0.13 mmol) and N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (72.0 mg, 0.19 mmol) and N,N- diisopropylethylamine (33.0 mg, 0.26 mmol). The reaction was stirred at 70 °C for 16 hours. The product was indicated via UPLC analysis. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified by prep-HPLC (the crude samples were dissolved in N,N-dimethylformamide before purification. Boston C1821*250mm 10µm column. The mobile phase was acetonitrile/0.01% aqueous formic acid) to give the product 1-cyclopropanecarbonyl-N- [6-(cyclopropylmethoxy)-1,3-benzothiazol-2-yl]-N-[(1H-imidazol-5-yl)methyl]piperidine-4-carboxamide (8.48 mg, 0.018 mmol, 14 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 12.00 (s, 1H), 7.67-7.70 (m, 1H), 7.59 (s, 1H), 7.49-7.50 (m, 1H), 7.08 (s, 1H), 7.01-7.04 (m, 1H), 5.49 (s, 2H), 4.30-4.42 (m, 2H), 3.85 (d, J = 7.2Hz, 2H), 3.60-3.67 (m, 1H), 3.14-3.26 (m, 1H), 2.62-2.67 (m, 1H), 1.98-2.01 (m, 1H), 1.80-1.88 (m, 2H), 1.44-1.62 (m, 2H), 1.21-1.27 (m, 1H), 0.70-0.71 (m, 4H), 0.55-0.60 (m, 2H), 0.31- 0.35 (m, 2H); LCMS (ESI) m/z: 480.3 [M+H]+. Example 244. Preparation of methyl 4-[(6-chloro-1,3-benzothiazol-2-yl)[(1H-imidazol-4- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000291_0001
Step 1: Preparation of 6-chloro-N-{[1-(triphenylmethyl)-1H-imidazol-4-yl]methyl}-1,3-benzothiazol-2-amine
Figure imgf000291_0002
In a reaction vial equipped with a magnetic stir bar, a solution of 1-(triphenylmethyl)-1H- imidazole-4-carbaldehyde (433 mg, 1.3 mmol) and 6-chloro-1,3-benzothiazol-2-amine (250 mg, 1.4 mmol) in dry toluene (7 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen for 24 hours. The reaction mixture was cooled to 60 °C and then to the reaction was added sodium borohydride (117 mg, 3.1 mmol) and dry methanol (7 mL). The reaction mixture was stirred at 64°C for 1 hour and then cooled to room temperature. The reaction was filtered over a bed of Celite and washed with methanol. The filtrate was concentrated then diluted with dichloromethane and re-filtered. The filtrate was purified via flash column chromatography eluting with 0-100% ethyl acetate in hexanes. Product 6-chloro- N-{[1-(triphenylmethyl)-1H-imidazol-4-yl]methyl}-1,3-benzothiazol-2-amine (100 mg, 0.20 mmol, 15 %) was afforded as a pale orange solid.1H NMR (300 MHz, Dimethylfulfoxide-d6) δ 8.38 (d, J = 5.5 Hz, 1H), 7.78 (t, J = 1.9 Hz, 1H), 7.51 – 7.28 (m, 7H), 7.22 (dt, J = 8.6, 2.0 Hz, 1H), 7.16 – 7.01 (m, 4H), 6.88 (d, J = 1.7 Hz, 1H), 4.41 (d, J = 5.1 Hz, 1H), 3.34 (d, J = 1.6 Hz, 5H); LCMS (ESI) m/z: 266.0 [M+H]+. Step 2: Preparation of methyl 4-[(6-chloro-1,3-benzothiazol-2-yl)({[1-(triphenylmethyl)-1H-imidazol-4- yl]methyl})carbamoyl]piperidine-1-carboxylate
Figure imgf000292_0001
In a reaction vial equipped with a stir bar, 6-chloro-N-{[1-(triphenylmethyl)-1H-imidazol-4- yl]methyl}-1,3-benzothiazol-2-amine (100 mg, 0.20 mmol) was dissolved in dichloromethane (1 mL). To the reaction solution was added 1-(methoxycarbonyl)piperidine-4-carboxylic acid (73.7 mg, 0.39 mmol) and 50% propylphosphonic anhydride in ethyl acetate (0.37 mL, 0.49 mmol). The reaction mixture was cooled to 0 °C and triethylamine (0.14 mL, 0.99 mmol) was added. The reaction was stirred from 0 °C to room temperature overnight. The product was indicated present via UPLC analysis. The reaction solution was diluted with dichloromethane and washed with water (3 x 15 mL). The organic layers were pooled, dried over anhydrous sodium sulfate, filtered and concentrated. Crude product was purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane. Product methyl 4-[(6- chloro-1,3-benzothiazol-2-yl)({[1-(triphenylmethyl)-1H-imidazol-4-yl]methyl})carbamoyl]piperidine-1- carboxylate (92.4 mg, 0.14 mmol, 69 %) was afforded as a white solid. Step 3: Preparation of methyl 4-[(6-chloro-1,3-benzothiazol-2-yl)[(1H-imidazol-4- yl)methyl]carbamoyl]piperidine-1-carboxylate
Figure imgf000292_0002
In a reaction vial equipped with a stir bar, methyl 4-[(6-chloro-1,3-benzothiazol-2-yl)({[1- (triphenylmethyl)-1H-imidazol-4-yl]methyl})carbamoyl]piperidine-1-carboxylate (92.4 mg, 0.14 mmol) was dissolved in dichloromethane (1 mL). To the reaction was added trifluoroacetic acid (0.083 mL, 1.1 mmol) and the reaction was stirred at room temperature for 2 hours. The product was indicated present via UPLC analysis. The reaction was concentrated under reduced pressure then diluted with dichloromethane and quenched with saturated sodium bicarbonate until bubbling ceased. The reaction was diluted with 10 mL deionized water and washed with dichloromethane (2x 10 mL). The organic layers were combined and dried over anhydrous sodium sulfate, filtered, then concentrated. Crude product was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane. Product methyl 4-[(6-chloro-1,3-benzothiazol-2-yl)[(1H-imidazol-4-yl)methyl]carbamoyl]piperidine-1-carboxylate (26.3 mg, 0.061 mmol, 45 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 7.91 – 7.63 (m, 3H), 7.42 (dd, J = 8.6, 2.1 Hz, 1H), 7.28 (d, J = 0.9 Hz, 1H), 7.17 (s, 1H), 5.52 (s, 2H), 4.24 (s, 3H), 3.73 (s, 3H), 3.50 (s, 1H), 2.99 (s, 2H), 1.83 (s, 4H); LCMS (ESI) m/z: 434.1 [M+H]+. Example 245. Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(1,3-thiazol-5- yl)methyl]propenamide
Figure imgf000293_0001
Step 1: Preparation of 5-fluoro-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000293_0002
In a reaction vial, a solution of 1,3-thiazole-5-carbaldehyde (201 mg, 1.8 mmol) and 5-fluoro-1,3- benzothiazol-2-amine (300 mg, 1.8 mmol) in dry toluene (18 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (154 mg, 4.1 mmol) in dry ethyl alcohol (18 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12- gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 5-fluoro-N-[(1,3- thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine (325 mg, 1.2 mmol, 69 %) was afforded as a orange solid. 1H NMR (300 MHz, Methanol-d4) δ 8.92 (s, 1H), 7.88 (s, 1H), 7.67 – 7.41 (m, 1H), 7.18 (dt, J = 10.2, 2.0 Hz, 1H), 6.86 (td, J = 8.9, 2.3 Hz, 1H), 4.89 (d, J = 1.3 Hz, 2H); LCMS (ESI) m/z: 265.9 [M+H]+. Step 2: Preparation of N-(5-fluoro-1,3-benzothiazol-2-yl)-3-phenoxy-N-[(1,3-thiazol-5- yl)methyl]propanamide
Figure imgf000293_0003
In a reaction vial, 5-fluoro-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine (172 mg, 0.65 mmol), 3-phenoxypropanoic acid (214 mg, 1.3 mmol), and triethylamine (0.45 mL, 3.2 mmol) were dissolved in dichloromethane (3 mL) with stirring. The reaction was cooled to 0 °C, and 50% propylphosphonic anhydride in ethyl acetate (1.2 mL, 1.6 mmol) was added. The reaction was warmed to room temperature and stirred for 2.5 days. The reaction was diluted with 10 mL deionized water and washed with 2x 10 mL portions of dichloromethane. The organic layers were separated, pooled, and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product N-(5-fluoro-1,3- benzothiazol-2-yl)-3-phenoxy-N-[(1,3-thiazol-5-yl)methyl]propanamide (22.5 mg, 0.054 mmol, 8.4 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 8.77 (s, 1H), 7.99 (s, 1H), 7.75 (dd, J = 8.7, 5.2 Hz, 1H), 7.57 (dd, J = 9.6, 2.4 Hz, 1H), 7.34 – 7.27 (m, 2H), 7.12 (td, J = 8.9, 2.5 Hz, 1H), 6.98 (tt, J = 7.3, 1.0 Hz, 1H), 6.97 – 6.87 (m, 2H), 5.77 (s, 2H), 4.44 (t, J = 6.4 Hz, 2H), 3.28 (t, J = 6.4 Hz, 2H); LCMS (ESI) m/z: 414.0 [M+H]+. Example 246. Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]oxane-4- carboxamide
Figure imgf000294_0001
Step 1: Preparation of 6-chloro-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000294_0002
In a reaction vial, a solution of pyrimidine-5-carbaldehyde (145 mg, 1.4 mmol) and 6-chloro-1,3- benzothiazol-2-amine (250 mg, 1.4 mmol) in dry toluene (14 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (117 mg, 3.1 mmol) in dry methanol (14 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-chloro-N-[(pyrimidin-5- yl)methyl]-1,3-benzothiazol-2-amine (129 mg, 0.47 mmol, 35 %) was afforded as a yellow solid.1H NMR (300 MHz, Methanol-d4) δ 9.07 (s, 1H), 8.85 (s, 1H), 8.78 (s, 1H), 7.63 (d, J = 2.2 Hz, 1H), 7.38 (d, J = 8.6 Hz, 1H), 7.24 (dd, J = 8.6, 2.2 Hz, 1H), 4.70 (s, 2H); LCMS (ESI) m/z: 276.9 [M+H]+. Step 2: Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]oxane-4-carboxamide
Figure imgf000294_0003
In a reaction vial, 6-chloro-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine (121 mg, 0.44 mmol) and N,N-diisopropylethylamine (0.23 mL, 1.3 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. oxane-4-carbonyl chloride (194 mg, 1.3 mmol) was added dropwise with stirring and the reaction was stirred for 16 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction. The layers were separated, and the organic layer was washed with 2 x 10 mL portions deionized water. The organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]oxane-4-carboxamide (104 mg, 0.27 mmol, 62 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 9.18 (s, 1H), 8.74 (s, 2H), 7.80 (d, J = 2.1 Hz, 1H), 7.68 (d, J = 8.6 Hz, 1H), 7.39 (dd, J = 8.7, 2.1 Hz, 1H), 5.56 (s, 2H), 4.15 – 3.79 (m, 2H), 3.63 – 3.31 (m, 2H), 2.97 (d, J = 12.5 Hz, 1H), 2.17 – 1.90 (m, 2H), 1.68 (d, J = 13.5 Hz, 2H); LCMS (ESI) m/z: 389.0 [M+H]+. Example 247. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5-yl)methyl]oxane-4- carboxamide
Figure imgf000295_0001
Step 1: Preparation of 6-fluoro-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000295_0002
In a reaction vial, a solution of 1,3-thiazole-5-carbaldehyde (242 mg, 2.1 mmol) and 6-fluoro-1,3- benzothiazol-2-amine (360 mg, 2.1 mmol) in dry toluene (21 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (186 mg, 4.9 mmol) in dry ethyl alcohol (21 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12- gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 6-fluoro-N-[(1,3- thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine (393 mg, 1.5 mmol, 69 %) was afforded as a white solid.1H NMR (300 MHz, Methanol-d4) δ 8.91 (s, 1H), 7.93 – 7.83 (m, 1H), 7.48 – 7.36 (m, 2H), 7.12 – 6.93 (m, 1H), 4.88 – 4.79 (m, 2H); LCMS (ESI) m/z: 266.0 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5-yl)methyl]oxane-4-carboxamide
Figure imgf000295_0003
In a reaction vial, 6-fluoro-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine (128 mg, 0.48 mmol) and N,N-diisopropylethylamine (0.25 mL, 1.4 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. oxane-4-carbonyl chloride (213 mg, 1.4 mmol) was added dropwise with stirring and the reaction was stirred for 16 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5-yl)methyl]oxane-4-carboxamide (90.8 mg, 0.24 mmol, 50 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.75 (s, 1H), 7.94 (s, 1H), 7.81 (dd, J = 8.9, 4.7 Hz, 1H), 7.51 (dd, J = 8.0, 2.6 Hz, 1H), 7.20 (td, J = 8.9, 2.6 Hz, 1H), 5.71 (s, 2H), 4.08 (d, J = 11.4 Hz, 2H), 3.49 (t, J = 11.7 Hz, 2H), 3.16 (tt, J = 11.3, 3.8 Hz, 1H), 2.16 – 1.92 (m, 2H), 1.70 (d, J = 13.9 Hz, 2H); LCMS (ESI) m/z: 378.0 [M+H]+. Example 248. Preparation of tert-butyl 4-{[(pyrimidin-5-yl)methyl][6-(trifluoromethyl)-1,3- benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000296_0001
Step 1: Preparation of N-[(pyrimidin-5-yl)methyl]-6-(trifluoromethyl)-1,3-benzothiazol-2-amine
Figure imgf000296_0002
In a reaction vial equipped with a magnetic stir bar, a solution of pyrimidine-5-carbaldehyde (140 mg, 1.3 mmol) and 6-(trifluoromethyl)-1,3-benzothiazol-2-amine (300 mg, 1.4 mmol) in dry toluene (7 mL) was stirred at 100 °C with activated 4 Å molecular sieves under nitrogen for 24 hours. The reaction mixture was cooled to 60 °C and then to the reaction was added sodium borohydride (129 mg, 3.4 mmol) and dry methanol (7 mL). The reaction mixture was stirred at 64°C for 1 hour and then cooled to room temperature. The reaction was filtered over a bed of Celite and washed with methanol. The filtrate was concentrated then diluted with dichloromethane and re-filtered. The filtrate was purified via flash column chromatography eluting with 0-20% methanol in dichloromethane. Product N-[(pyrimidin-5-yl)methyl]-6- (trifluoromethyl)-1,3-benzothiazol-2-amine (182 mg, 0.59 mmol, 43 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 9.20 (s, 1H), 8.86 (s, 2H), 7.87 (dt, J = 1.6, 0.8 Hz, 1H), 7.72 – 7.46 (m, 2H), 5.93 (s, 1H), 4.77 (s, 2H); LCMS (ESI) m/z: 311.0 [M+H]+. Step 2: Preparation of tert-butyl 4-{[(pyrimidin-5-yl)methyl][6-(trifluoromethyl)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000296_0003
In a reaction vial equipped with a stir bar, N-[(pyrimidin-5-yl)methyl]-6-(trifluoromethyl)-1,3- benzothiazol-2-amine (182 mg, 0.59 mmol) was dissolved in dichloromethane (3 mL). To the reaction solution was added 1-[(tert-butoxy)carbonyl]piperidine-4-carboxylic acid (268 mg, 1.2 mmol) and 50% propylphosphonic anhydride in ethyl acetate (1.10 mL, 1.5 mmol). The reaction mixture was cooled to 0 °C and triethylamine (0.41 mL, 2.9 mmol) was added. The reaction was stirred from 0 °C to room temperature overnight. The product was indicated present via UPLC analysis. The reaction solution was diluted with dichloromethane and washed with water (3 x 15 mL). The organic layers were pooled, dried over anhydrous sodium sulfate, filtered and concentrated. Crude product was purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane. Product tert-butyl 4-{[(pyrimidin-5- yl)methyl][6-(trifluoromethyl)-1,3-benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate (91.4 mg, 0.18 mmol, 30 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 9.21 (s, 1H), 8.76 (s, 2H), 8.13 (dd, J = 1.9, 0.9 Hz, 1H), 7.86 (d, J = 8.5 Hz, 1H), 7.77 – 7.63 (m, 1H), 5.62 (s, 2H), 4.20 (s, 2H), 3.00 – 2.65 (m, 3H), 2.01 – 1.64 (m, 4H), 1.48 (s, 9H); LCMS (ESI) m/z: 422.1 [M-t-butyl]+. Example 249. Preparation of N-[(2S)-5-methoxy-2,3-dihydro-1H-inden-2-yl]-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000297_0001
Step 1: Preparation of 5-methoxy-N-(pyridin-3-ylmethyl)-2,3-dihydro-1H-inden-2-amine
Figure imgf000297_0002
To a solution of 5-methoxy-2,3-dihydro-1H-inden-2-amine (100 mg, 0.61 mmol) and nicotinaldehyde (72.0 mg, 0.67 mmol) in toluene (8 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (70.0 mg, 1.8 mmol) in methanol (10 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane. Product 5-methoxy-N-(pyridin-3-ylmethyl)-2,3-dihydro-1H-inden-2-amine (80.0 mg, 0.32 mmol, 52 %) as a white solid. LCMS (ESI) m/z: 255.1 [M+H]+. Step 2: Preparation of N-[(2S)-5-methoxy-2,3-dihydro-1H-inden-2-yl]-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000297_0003
In a reaction vial, a mixture of 5-methoxy-N-(pyridin-3-ylmethyl)-2,3-dihydro-1H-inden-2-amine (45.0 mg, 0.35 mmol), N,N-diisopropylethylamine (92.0 mg, 0.71 mmol) and N-[(dimethylamino)-1H-1,2,3- triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (135 mg, 0.35 mmol) in tetrahydrofuran (10 mL) was stirred at room temperature for 30 minutes. Then, cyclohexanecarboxylic acid (60.0 mg, 0.24 mmol) was added, and the reaction was stirred at 70 °C for 16 hours. The product was indicated present via UPLC analysis. Water (20 mL) was added to the reaction mixture, and the organics were extracted with 3x 20 mL portions of ethyl acetate. The organic layers were pooled, dried over sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane. The racemic product was separated via chiral-HPLC to give N-[(2S)-5-methoxy-2,3- dihydro-1H-inden-2-yl]-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide (6.40 mg, 0.018 mmol, 5.0 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.42-8.29 (m, 2H), 7.47-7.30 (m, 2H), 7.03 (d, J = 4.0 Hz, 2H), 6.74 (s, 1H), 6.69 (d, J = 4.0 Hz, 2H), 5.01 (t, J = 7.6 Hz, 1H), 4.53 (s, 2H), 3.52 (s, 3H), 3.18 - 2.80 (m, 4H), 1.70- 1.23 (m, 11H); LCMS (ESI) m/z: 365.3 [M+H]+. Example 250. Preparation of N-[(2R)-5-methoxy-2,3-dihydro-1H-inden-2-yl]-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000298_0001
Step 1: Preparation of 5-methoxy-N-(pyridin-3-ylmethyl)-2,3-dihydro-1H-inden-2-amine
Figure imgf000298_0002
To a solution of 5-methoxy-2,3-dihydro-1H-inden-2-amine (100 mg, 0.61 mmol) and nicotinaldehyde (72.0 mg, 0.67 mmol) in toluene (8 mL) was added dried 4 Å molecular sieves. The mixture was heated to 110 °C and stirred for 16 hours. Then a solution of sodium borohydride (70.0 mg, 1.8 mmol) in methanol (10 mL) was added. The reaction was stirred at room temperature for 2 hours. The reaction was filtered over Celite and the solvent was removed under the reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane. Product 5-methoxy-N-(pyridin-3-ylmethyl)-2,3-dihydro-1H-inden-2-amine (80.0 mg, 0.32 mmol, 52 %) as a white solid. LCMS (ESI) m/z: 255.1 [M+H]+. Step 2: Preparation of N-[(2R)-5-methoxy-2,3-dihydro-1H-inden-2-yl]-N-[(pyridin-3- yl)methyl]cyclohexanecarboxamide
Figure imgf000298_0003
In a reaction vial, a mixture of 5-methoxy-N-(pyridin-3-ylmethyl)-2,3-dihydro-1H-inden-2-amine (45.0 mg, 0.35 mmol), N,N-diisopropylethylamine (92.0 mg, 0.71 mmol) and N-[(dimethylamino)-1H-1,2,3- triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (135 mg, 0.35 mmol) in tetrahydrofuran (10 mL) was stirred at room temperature for 30 minutes. Then, cyclohexanecarboxylic acid (60.0 mg, 0.24 mmol) was added, and the reaction was stirred at 70 °C for 16 hours. The product was indicated present via UPLC analysis. Water (20 mL) was added to the reaction mixture, and the organics were extracted with 3x 20 mL portions of ethyl acetate. The organic layers were pooled, dried over sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified via flash column chromatography through silica gel using a gradient of 0-5% methanol in dichloromethane. The racemic product was separated via chiral-HPLC to give N-[(2R)-5-methoxy-2,3- dihydro-1H-inden-2-yl]-N-[(pyridin-3-yl)methyl]cyclohexanecarboxamide (5.60 mg, 0.015 mmol, 4.4 %) as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 8.42-8.29 (m, 2H), 7.47-7.30 (m, 2H), 7.03 (d, J = 4.0 Hz, 2H), 6.74 (s, 1H), 6.69 (d, J = 4.0 Hz, 2H), 5.01 (t, J = 7.6 Hz, 1H), 4.53 (s, 2H), 3.52 (s, 3H), 3.18 - 2.80 (m, 4H), 1.70- 1.23 (m, 11H); LCMS (ESI) m/z: 365.4 [M+H]+. Example 251. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-oxazol-5-yl)methyl]oxane-4- carboxamide
Figure imgf000299_0001
Step 1: Preparation of 6-fluoro-N-[(1,3-oxazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000299_0002
In a reaction vial, a solution of 1,3-oxazole-5-carbaldehyde (114 mg, 1.2 mmol) and 6-fluoro-1,3- benzothiazol-2-amine (200 mg, 1.2 mmol) in dry toluene (12 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry methanol (12 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-fluoro-N-[(1,3-oxazol-5- yl)methyl]-1,3-benzothiazol-2-amine (145 mg, 0.58 mmol, 49 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 7.85 (s, 1H), 7.50 (dd, J = 8.9, 4.7 Hz, 1H), 7.30 (dd, J = 8.1, 2.6 Hz, 1H), 7.10 (s, 1H), 7.11 – 6.97 (m, 1H), 4.73 (d, J = 0.9 Hz, 2H); LCMS (ESI) m/z: 249.9 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-oxazol-5-yl)methyl]oxane-4-carboxamide
Figure imgf000299_0003
In a reaction vial, 6-fluoro-N-[(1,3-oxazol-5-yl)methyl]-1,3-benzothiazol-2-amine (139 mg, 0.56 mmol) and N,N-diisopropylethylamine (0.29 mL, 1.7 mmol) were dissolved in dimethylformamide (1.5 mL) and cooled to 0 °C. oxane-4-carbonyl chloride (248 mg, 1.7 mmol) was added dropwise with stirring and the reaction was stirred for 16 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-oxazol-5-yl)methyl]oxane-4-carboxamide (14.7 mg, 0.041 mmol, 7.31 %) was afforded as a white solid.1H NMR (300 MHz, Acetone-d6) δ 8.10 (s, 1H), 7.84 (dd, J = 8.9, 4.7 Hz, 1H), 7.76 (dd, J = 8.5, 2.7 Hz, 1H), 7.28 (dd, J = 9.1, 2.7 Hz, 1H), 7.23 (d, J = 1.9 Hz, 1H), 5.80 (s, 2H), 3.97 (dd, J = 10.2, 2.8 Hz, 2H), 3.56 (qd, J = 11.4, 10.8, 4.0 Hz, 4H), 1.87 (dd, J = 10.9, 4.1 Hz, 4H); LCMS (ESI) m/z: 362.1 [M+H]+. Example 252. Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5-yl)methyl]oxane-4- carboxamide
Figure imgf000300_0001
Step 1: Preparation of 4-fluoro-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000300_0002
In a reaction vial, a solution of 1,3-thiazole-5-carbaldehyde (201 mg, 1.8 mmol) and 4-fluoro-1,3- benzothiazol-2-amine (300 mg, 1.8 mmol) in dry toluene (18 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (154 mg, 4.1 mmol) in dry methanol (18 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 4-fluoro-N-[(1,3-thiazol-5- yl)methyl]-1,3-benzothiazol-2-amine (322 mg, 1.2 mmol, 68 %) was afforded as an orange solid.1H NMR (300 MHz, Chloroform-d) δ 8.76 (d, J = 0.7 Hz, 1H), 7.90 (d, J = 0.8 Hz, 1H), 7.41 – 7.33 (m, 1H), 7.10 – 7.01 (m, 2H), 4.92 (s, 2H); LCMS (ESI) m/z: 265.9 [M+H]+. Step 2: Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5-yl)methyl]oxane-4-carboxamide
Figure imgf000300_0003
In a reaction vial, 4-fluoro-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine (170 mg, 0.64 mmol) and N,N-diisopropylethylamine (0.33 mL, 1.9 mmol) were dissolved in dimethylformamide (1.5 mL) and cooled to 0 °C. oxane-4-carbonyl chloride (282 mg, 1.9 mmol) was added dropwise with stirring and the reaction was stirred for 16 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-(4-fluoro-1,3-benzothiazol-2-yl)-N-[(1,3-thiazol-5-yl)methyl]oxane-4-carboxamide (105 mg, 0.28 mmol, 44 %) was afforded as a white solid.1H NMR (300 MHz, Acetone-d6) δ 8.99 (d, J = 0.8 Hz, 1H), 8.39 – 8.04 (m, 1H), 7.84 (dd, J = 7.9, 1.0 Hz, 1H), 7.44 (td, J = 8.0, 4.7 Hz, 1H), 7.33 (ddd, J = 10.8, 8.0, 1.0 Hz, 1H), 6.11 – 5.93 (m, 2H), 4.04 (ddd, J = 11.6, 4.4, 2.0 Hz, 2H), 3.73 – 3.52 (m, 3H), 2.06 – 1.76 (m, 4H); LCMS (ESI) m/z: 378.0 [M+H]+. Example 253. Preparation of N-[(1,3-thiazol-5-yl)methyl]-N-[6-(trifluoromethoxy)-1,3-benzothiazol-2- yl]oxane-4-carboxamide
Figure imgf000301_0001
Step 1: Preparation of N-(thiazol-5-ylmethyl)-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine
Figure imgf000301_0002
To a solution of thiazole-5-carbaldehyde (3.00 g, 27 mmol) and 6-(trifluoromethoxy)-1,3- benzothiazol-2-amine (4.00 g, 17 mmol) in dry diglyme (50 mL) was added titanium isopropoxide (9.71 g, 34 mmol) in a flamed dried flask. The mixture was heated to 60 °C with stirring for 1.5 hours. Then, the mixture was cooled to 0 °C and sodium borohydride (0.969 g, 26 mmol) was added. The reaction was warmed to room temperature over 2 hours. The product was indicated via UPLC analysis. The reaction was quenched with 10 mL water, and the organics were extracted with ethyl acetate (5x 30 mL). The organic layers were pooled, washed with brine (3x 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by reversed-phase HPLC (30% methanol in water solution condition) to give N-(thiazol-5-ylmethyl)-6-(trifluoromethoxy)-1,3-benzothiazol- 2-amine (2.41 g, 7.1 mmol, 42 %) as a white solid.1H NMR (400 MHz, Chloroform-d) δ 8.75 (s, 1H), 7.87 (s, 1H), 7.52 (d, J=8.8 Hz, 1H), 7.46 (d, J=1.1 Hz, 1H), 7.18 (dd, J=1.4, 8.7 Hz, 1H), 5.70 (br s, 1H), 5.01 (s, 2H); LCMS (ESI) m/z: 332.0 [M+H]+. Step 2: Preparation of N-[(1,3-thiazol-5-yl)methyl]-N-[6-(trifluoromethoxy)-1,3-benzothiazol-2-yl]oxane-4- carboxamide
Figure imgf000301_0003
In a reaction vial, N-[(1,3-thiazol-5-yl)methyl]-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (150 mg, 0.45 mmol) and N,N-diisopropylethylamine (0.23 mL, 1.4 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. oxane-4-carbonyl chloride (200 mg, 1.4 mmol) was added dropwise with stirring and the reaction was stirred for 3 hours. The product was confirmed present via UPLC analysis. 10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-50% ethyl acetate in dichloromethane. Product N-[(1,3-thiazol-5-yl)methyl]-N-[6-(trifluoromethoxy)-1,3-benzothiazol-2-yl]oxane- 4-carboxamide (147 mg, 0.33 mmol, 73 %) was afforded as a white solid.1H NMR (300 MHz, Acetone-d6) δ 8.91 (d, J = 0.8 Hz, 1H), 8.12 (q, J = 0.8 Hz, 1H), 8.03 (dd, J = 2.4, 1.2 Hz, 1H), 7.95 (d, J = 8.8 Hz, 1H), 7.53 – 7.39 (m, 1H), 5.97 (d, J = 1.0 Hz, 2H), 3.95 (ddd, J = 11.6, 4.5, 2.0 Hz, 2H), 3.56 (tt, J = 11.7, 2.8 Hz, 3H), 1.95 – 1.64 (m, 4H); LCMS (ESI) m/z: 444.0 [M+H]+. Example 254. Preparation of N-(4-fluoro-1H-1,3-benzodiazol-2-yl)-N-[(1,3-thiazol-5-yl)methyl]oxane- 4-carboxamide
Figure imgf000302_0001
Step 1: Preparation of 4-fluoro-N-[(1,3-thiazol-5-yl)methyl]-1H-1,3-benzodiazol-2-amine
Figure imgf000302_0002
In a small microwave reaction flask, 2-chloro-4-fluoro-1H-1,3-benzodiazole (200 mg, 1.2 mmol), 1-(1,3-thiazol-5-yl)methanamine hydrochloride (176 mg, 1.2 mmol) and N,N-diisopropylethylamine (0.22 mL, 1.3 mmol) were dissolved in acetonitrile (4 mL). The reaction was microwaved for 30 minutes at 170 °C. The reaction was cooled to room temperature and the solvent was evaporated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-50% ethyl acetate in dichloromethane. Product 4-fluoro-N-[(1,3-thiazol-5-yl)methyl]- 1H-1,3-benzodiazol-2-amine (73.8 mg, 0.30 mmol, 25 %) was afforded as a yellow solid.1H NMR (300 MHz, Methanol-d4) δ 8.90 (s, 1H), 7.86 (s, 1H), 7.14 – 6.96 (m, 1H), 6.93 (tdd, J = 8.1, 4.8, 1.3 Hz, 1H), 6.84 – 6.67 (m, 1H), 4.83 (d, J = 1.5 Hz, 2H); LCMS (ESI) m/z: 248.9 [M+H]+. Step 2: Preparation of N-(4-fluoro-1H-1,3-benzodiazol-2-yl)-N-[(1,3-thiazol-5-yl)methyl]oxane-4- carboxamide
Figure imgf000302_0003
In a reaction vial, 4-fluoro-N-[(1,3-thiazol-5-yl)methyl]-1H-1,3-benzodiazol-2-amine (66.1 mg, 0.27 mmol) and N,N-diisopropylethylamine (0.14 mL, 0.80 mmol) were dissolved in tetrahydrofuran (1 mL) and cooled to 0 °C. oxane-4-carbonyl chloride (94.8 mg, 0.64 mmol) was added dropwise with stirring and the reaction was stirred overnight. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-(4-fluoro-1H-1,3-benzodiazol-2-yl)-N-[(1,3-thiazol-5-yl)methyl]oxane-4-carboxamide (22.0 mg, 0.061 mmol, 23 %) was afforded as a yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.77 (s, 1H), 7.96 (s, 1H), 7.33 – 7.22 (m, 1H), 7.24 – 7.11 (m, 1H), 6.99 (dd, J = 10.5, 8.3 Hz, 1H), 5.76 (s, 2H), 4.09 (d, J = 11.9 Hz, 2H), 3.50 (t, J = 11.7 Hz, 2H), 3.22 – 3.04 (m, 1H), 2.02 (qd, J = 12.7, 11.7, 4.4 Hz, 2H), 1.67 (d, J = 13.5 Hz, 2H); LCMS (ESI) m/z: 361.1 [M+H]+. Example 255. Preparation of N-[(pyrimidin-5-yl)methyl]-N-[6-(trifluoromethoxy)-1,3-benzothiazol-2- yl]oxane-4-carboxamide
Figure imgf000303_0001
Step 1: Preparation of N-(pyrimidin-5-ylmethyl)-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine
Figure imgf000303_0002
To a solution of pyrimidine-5-carbaldehyde (1.94 g, 18 mmol) and 6-(trifluoromethyl)-1,3- benzothiazol-2-amine (3.51 g, 15 mmol) in tetrahydrofuran (50 mL) was added titanium isopropoxide (10.6 g, 38 mmol) in a flamed dried flask. The mixture was heated to 60 °C with stirring for 2 hours. Then, the mixture was cooled to 0 °C and sodium borohydride (0.851 g, 23 mmol) was added. The reaction was warmed to room temperature over 2 hours. The product was indicated via UPLC analysis. The reaction was quenched with 10 mL water, and the organics were extracted with ethyl acetate (5x 30 mL). The organic layers were pooled, washed with brine (3x 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by flash column chromatography through 80 grams of silica gel using a gradient of 0-30 % ethyl acetate in petroleum ether. Product N- (pyrimidin-5-ylmethyl)-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (1.76 g, 5.4 mmol, 36 %) was obtained as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 9.10 (s, 1H), 8.84(s, 2H), 8.75-8.72 (m, 1H), 7.82 (d, 1H), 7.47 (d, J = 8.8 Hz, 1H), 7.21-7.19 (dd, J = 1.6, 8.8 Hz, 1H), 4.65 (d, J = 5.6, 2H); LCMS (ESI) m/z: 327.0 [M+H]+. Step 2: Preparation of N-[(pyrimidin-5-yl)methyl]-N-[6-(trifluoromethoxy)-1,3-benzothiazol-2-yl]oxane-4- carboxamide
Figure imgf000303_0003
In a reaction vial, N-[(pyrimidin-5-yl)methyl]-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (150 mg, 0.46 mol) and N,N-diisopropylethylamine (0.24 mL, 1.4 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. oxane-4-carbonyl chloride (203 mg, 1.4 mmol) was added dropwise with stirring and the reaction was stirred for 3 hours. The product was confirmed present via UPLC analysis. 10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product N-[(pyrimidin-5-yl)methyl]-N-[6-(trifluoromethoxy)-1,3-benzothiazol-2-yl]oxane- 4-carboxamide (164 mg, 0.38 mmol, 82 %) was afforded as a white solid.1H NMR (300 MHz, Acetone-d6) δ 9.06 (s, 1H), 8.84 (d, J = 0.8 Hz, 2H), 8.01 (dd, J = 2.4, 1.2 Hz, 1H), 7.83 (d, J = 8.8 Hz, 1H), 7.40 (ddt, J = 8.9, 2.6, 0.9 Hz, 1H), 5.82 (s, 2H), 3.93 (ddd, J = 11.5, 4.3, 2.1 Hz, 2H), 3.47 (td, J = 11.6, 2.6 Hz, 3H), 1.99 – 1.71 (m, 4H); LCMS (ESI) m/z: 439.0 [M+H]+. Example 256. Preparation of N-[(pyrimidin-5-yl)methyl]-N-[6-(trifluoromethyl)-1,3-benzothiazol-2- yl]oxane-4-carboxamide
Figure imgf000304_0001
Step 1: Preparation of N-(pyrimidin-5-ylmethyl)-6-(trifluoromethyl)-1,3-benzothiazol-2-amine
Figure imgf000304_0002
To a solution of pyrimidine-5-carbaldehyde (2.03 g, 19 mmol) and 6-(trifluoromethyl)-1,3- benzothiazol-2-amine (3.42 g, 16 mmol) in tetrahydrofuran (50 mL) was added titanium isopropoxide (11.1 g, 39 mmol) in a flamed dried flask. The mixture was heated to 60 °C with stirring for 2 hours. Then, the mixture was cooled to 0 °C and sodium borohydride (0.889 g, 24 mmol) was added. The reaction was warmed to room temperature over 2 hours. The product was indicated via UPLC analysis. The reaction was quenched with 10 mL water, and the organics were extracted with ethyl acetate (5x 30 mL). The organic layers were pooled, washed with brine (3x 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by flash column chromatography through 80 grams of silica gel using a gradient of 0-30 % ethyl acetate in petroleum ether. Product N- (pyrimidin-5-ylmethyl)-6-(trifluoromethyl)-1,3-benzothiazol-2-amine (1.75 g, 5.6 mmol, 36 %) was obtained as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 9.11 (s, 1H), 8.93-8.90 (m, 1H), 8.84 (s, 2H), 8.15 (s, 1H), 7.56-7.51 (m, 2H), 4.69-4.67 (m, 2H); LCMS (ESI) m/z: 311.0 [M+H]+. Step 2: Preparation of N-[(pyrimidin-5-yl)methyl]-N-[6-(trifluoromethyl)-1,3-benzothiazol-2-yl]oxane-4- carboxamide
Figure imgf000304_0003
In a reaction vial, N-[(pyrimidin-5-yl)methyl]-6-(trifluoromethyl)-1,3-benzothiazol-2-amine (150 mg, 0.48 mmol) and N,N-diisopropylethylamine (0.25 mL, 1.4 mmol) were dissolved in dimethylformamide (1.5 mL) and cooled to 0 °C. oxane-4-carbonyl chloride (213 mg, 1.4 mmol) was added dropwise with stirring and the reaction was stirred for 3 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product N-[(pyrimidin-5-yl)methyl]-N-[6-(trifluoromethyl)-1,3-benzothiazol-2-yl]oxane-4-carboxamide (63.0 mg, 0.15 mmol, 31 %) was afforded as a white solid.1H NMR (300 MHz, Acetone-d6) δ 9.07 (s, 1H), 8.85 (d, J = 0.8 Hz, 2H), 8.40 (q, J = 1.2, 0.8 Hz, 1H), 7.91 (dt, J = 8.5, 0.8 Hz, 1H), 7.79 – 7.69 (m, 1H), 5.85 (s, 2H), 3.93 (dd, J = 10.5, 3.8 Hz, 2H), 3.47 (td, J = 11.8, 2.5 Hz, 3H), 1.87 (dt, J = 21.7, 12.4 Hz, 4H); LCMS (ESI) m/z: 423.1 [M+H]+. Example 257. Preparation of N-[(4-methylpyridin-3-yl)methyl]-N-[6-(trifluoromethyl)-1,3- benzothiazol-2-yl]oxane-4-carboxamide
Figure imgf000305_0001
Step 1: Preparation of N-[(4-methyl-3-pyridyl)methyl]-6-(trifluoromethyl)-1,3-benzothiazol-2-amine
Figure imgf000305_0002
To a solution of 4-methylpyridine-3-carbaldehyde (2.02 g, 17 mmol) and 6-(trifluoromethyl)-1,3- benzothiazol-2-amine (3.05 g, 14 mmol) in tetrahydrofuran (50 mL) was added titanium isopropoxide (9.94 g, 35 mmol) in a flamed dried flask. The mixture was heated to 60 °C with stirring for 2 hours. Then, the mixture was cooled to 0 °C and sodium borohydride (0.794 g, 21 mmol) was added. The reaction was warmed to room temperature over 2 hours. The product was indicated via UPLC analysis. The reaction was quenched with 10 mL water, and the organics were extracted with ethyl acetate (5x 30 mL). The organic layers were pooled, washed with brine (3x 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by flash column chromatography through 80 grams of silica gel using a gradient of 0-30 % ethyl acetate in petroleum ether. Product N-[(4- methyl-3-pyridyl)methyl]-6-(trifluoromethyl)-1,3-benzothiazol-2-amine (1.26 g, 3.9 mmol, 28 %) was obtained as a white solid.1H NMR (400 MHz, Chloroform-d) δ 8.50 (s, 1H), 8.43 (d, J = 5.0 Hz, 1H), 7.85 (s, 1H), 7.55 (s, 2H), 7.13 (d, J = 5.0 Hz, 1H), 6.24 (br s, 1H), 4.71 (s, 2H), 2.41 (s, 3H); LCMS (ESI) m/z: 324.1 [M+H]+. Step 2: Preparation of N-[(4-methylpyridin-3-yl)methyl]-N-[6-(trifluoromethyl)-1,3-benzothiazol-2-yl]oxane- 4-carboxamide
Figure imgf000305_0003
In a reaction vial, N-[(4-methylpyridin-3-yl)methyl]-6-(trifluoromethyl)-1,3-benzothiazol-2-amine (150 mg, 0.46 mmol) and N,N-diisopropylethylamine (0.24 mL, 1.4 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. oxane-4-carbonyl chloride (205 mg, 1.4 mmol) was added dropwise with stirring and the reaction was stirred for 3 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product N-[(4-methylpyridin-3-yl)methyl]-N-[6-(trifluoromethyl)-1,3-benzothiazol- 2-yl]oxane-4-carboxamide (169 mg, 0.39 mmol, 84 %) was afforded as a white solid.1H NMR (300 MHz, Acetone-d6) δ 8.41 (dt, J = 1.8, 0.9 Hz, 1H), 8.34 (d, J = 4.9 Hz, 1H), 8.07 (s, 1H), 7.84 (dt, J = 8.6, 0.8 Hz, 1H), 7.71 (dd, J = 8.7, 1.9 Hz, 1H), 7.26 (dt, J = 4.9, 0.8 Hz, 1H), 5.82 (s, 2H), 3.91 (ddd, J = 11.6, 4.5, 1.9 Hz, 2H), 3.37 (td, J = 11.8, 2.3 Hz, 2H), 3.25 (tt, J = 11.2, 3.9 Hz, 1H), 2.53 (s, 3H), 2.00 – 1.83 (m, 2H), 1.76 (dq, J = 13.2, 2.1 Hz, 2H); LCMS (ESI) m/z: 436.1 [M+H]+. Example 258. Preparation of methyl 4-{[(pyrimidin-5-yl)methyl][6-(trifluoromethyl)-1,3- benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000306_0001
Step 1: Preparation of N-(pyrimidin-5-ylmethyl)-6-(trifluoromethyl)-1,3-benzothiazol-2-amine
Figure imgf000306_0002
To a solution of pyrimidine-5-carbaldehyde (2.03 g, 19 mmol) and 6-(trifluoromethyl)-1,3- benzothiazol-2-amine (3.42 g, 16 mmol) in tetrahydrofuran (50 mL) was added titanium isopropoxide (11.1 g, 39 mmol) in a flamed dried flask. The mixture was heated to 60 °C with stirring for 2 hours. Then, the mixture was cooled to 0 °C and sodium borohydride (0.889 g, 24 mmol) was added. The reaction was warmed to room temperature over 2 hours. The product was indicated via UPLC analysis. The reaction was quenched with 10 mL water, and the organics were extracted with ethyl acetate (5x 30 mL). The organic layers were pooled, washed with brine (3x 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by flash column chromatography through 80 grams of silica gel using a gradient of 0-30 % ethyl acetate in petroleum ether. Product N- (pyrimidin-5-ylmethyl)-6-(trifluoromethyl)-1,3-benzothiazol-2-amine (1.75 g, 5.6 mmol, 36 %) was obtained as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 9.11 (s, 1H), 8.93-8.90 (m, 1H), 8.84 (s, 2H), 8.15 (s, 1H), 7.56-7.51 (m, 2H), 4.69-4.67 (m, 2H); LCMS (ESI) m/z: 311.0 [M+H]+. Step 2: Preparation of methyl 4-{[(pyrimidin-5-yl)methyl][6-(trifluoromethyl)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000306_0003
In a reaction vial equipped with a stir bar, N-[(pyrimidin-5-yl)methyl]-6-(trifluoromethyl)-1,3- benzothiazol-2-amine (250 mg, 0.81 mmol) was dissolved in dichloromethane (4 mL). To the reaction solution was added 1-(methoxycarbonyl)piperidine-4-carboxylic acid (299 mg, 1.6 mmol) and 50% propylphosphonic anhydride in ethyl acetate (1.5 mL, 2.0 mmol). The reaction mixture was cooled to 0 °C and triethylamine (0.56 mL, 4.0 mmol) was added. The reaction was stirred from 0 °C to room temperature overnight. The product was indicated present via UPLC analysis. The reaction solution was diluted with dichloromethane and washed with water (3x 15 mL). The organic layers were pooled, dried over anhydrous sodium sulfate, filtered and concentrated. The product was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane. Product methyl 4-{[(pyrimidin-5- yl)methyl][6-(trifluoromethyl)-1,3-benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate (107 mg, 0.22 mmol, 28 %) was afforded as an orange solid.1H NMR (300 MHz, Chloroform-d) δ 9.21 (s, 1H), 8.77 (s, 2H), 8.13 (dt, J = 1.7, 0.9 Hz, 1H), 7.86 (d, J = 8.5 Hz, 1H), 7.77 – 7.63 (m, 1H), 5.62 (s, 2H), 4.16 (d, J = 51.6 Hz, 3H), 2.90 (dt, J = 26.7, 12.2 Hz, 4H), 2.03 – 1.54 (m, 6H), 1.24 (d, J = 13.1 Hz, 2H); LCMS (ESI) m/z: 480.1 [M+H]+. Example 259. Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-1-propanoyl-N-[(1,3-thiazol-5- yl)methyl]piperidine-4-carboxamide
Figure imgf000307_0001
Step 1: Preparation of 4-fluoro-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000307_0002
In a reaction vial, a solution of 1,3-thiazole-5-carbaldehyde (201 mg, 1.8 mmol) and 4-fluoro-1,3- benzothiazol-2-amine (300 mg, 1.8 mmol) in dry toluene (18 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (154 mg, 4.1 mmol) in dry methanol (18 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12 gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 4-fluoro-N-[(1,3-thiazol-5- yl)methyl]-1,3-benzothiazol-2-amine (322 mg, 1.2 mmol, 68 %) was afforded as an orange solid.1H NMR (300 MHz, Chloroform-d) δ 8.76 (d, J = 0.7 Hz, 1H), 7.90 (d, J = 0.8 Hz, 1H), 7.41 – 7.33 (m, 1H), 7.10 – 7.01 (m, 2H), 4.92 (s, 2H); LCMS (ESI) m/z: 265.9 [M+H]+. Step 2: Preparation of N-(4-fluoro-1,3-benzothiazol-2-yl)-1-propanoyl-N-[(1,3-thiazol-5- yl)methyl]piperidine-4-carboxamide
Figure imgf000307_0003
In a reaction vial, 4-fluoro-N-[(1,3-thiazol-5-yl)methyl]-1,3-benzothiazol-2-amine (149 mg, 0.56 mmol), 1-propanoylpiperidine-4-carboxylic acid (207 mg, 1.1 mmol), and 50% propylphosphonic anhydride in ethyl acetate (1.05 mL, 1.4 mmol) were dissolved in dichloromethane (3 mL) with stirring. The reaction was cooled to 0 °C, and triethylamine (0.39 mL, 2.8 mmol) was added. The reaction was warmed to room temperature and stirred for 2.5 days. The reaction was diluted with 10 mL deionized water and washed with 2x 10 mL portions of dichloromethane. The organic layers were separated, pooled, and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product N-(4-fluoro-1,3-benzothiazol-2-yl)-1-propanoyl-N-[(1,3-thiazol-5-yl)methyl]piperidine-4- carboxamide (84.9 mg, 0.20 mmol, 35 %) was afforded as a white solid. 1H NMR (300 MHz, Acetone-d6) δ 8.92 (s, 1H), 8.22 – 8.09 (m, 1H), 7.79 (dd, J = 7.9, 1.0 Hz, 1H), 7.38 (td, J = 8.0, 4.6 Hz, 1H), 7.26 (ddd, J = 10.8, 8.1, 1.0 Hz, 1H), 5.99 (s, 2H), 4.61 (d, J = 13.3 Hz, 1H), 4.12 – 3.95 (m, 1H), 3.61 (tt, J = 11.2, 3.8 Hz, 1H), 3.27 (t, J = 13.0 Hz, 1H), 2.83 – 2.67 (m, 1H), 2.44 – 2.28 (m, 2H), 1.89 (d, J = 12.8 Hz, 2H), 1.83 – 1.54 (m, 2H), 1.06 (t, J = 7.4 Hz, 3H); LCMS (ESI) m/z: 433.0 [M+H]+. Example 260. Preparation of methyl 4-{[(4-methylpyridin-3-yl)methyl][6-(trifluoromethyl)-1,3- benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000308_0001
Step 1: Preparation of N-[(4-methyl-3-pyridyl)methyl]-6-(trifluoromethyl)-1,3-benzothiazol-2-amine
Figure imgf000308_0002
To a solution of 4-methylpyridine-3-carbaldehyde (2.02 g, 17 mmol) and 6-(trifluoromethyl)-1,3- benzothiazol-2-amine (3.05 g, 14 mmol) in tetrahydrofuran (50 mL) was added titanium isopropoxide (9.94 g, 35 mmol) in a flamed dried flask. The mixture was heated to 60 °C with stirring for 2 hours. Then, the mixture was cooled to 0 °C and sodium borohydride (0.794 g, 21 mmol) was added. The reaction was warmed to room temperature over 2 hours. The product was indicated via UPLC analysis. The reaction was quenched with 10 mL water, and the organics were extracted with ethyl acetate (5x 30 mL). The organic layers were pooled, washed with brine (3x 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by flash column chromatography through 80 grams of silica gel using a gradient of 0-30 % ethyl acetate in petroleum ether. Product N-[(4- methyl-3-pyridyl)methyl]-6-(trifluoromethyl)-1,3-benzothiazol-2-amine (1.26 g, 3.9 mmol, 28 %) was obtained as a white solid.1H NMR (400 MHz, Chloroform-d) δ 8.50 (s, 1H), 8.43 (d, J = 5.0 Hz, 1H), 7.85 (s, 1H), 7.55 (s, 2H), 7.13 (d, J = 5.0 Hz, 1H), 6.24 (br s, 1H), 4.71 (s, 2H), 2.41 (s, 3H); LCMS (ESI) m/z: 324.1 [M+H]+. Step 2: Preparation of methyl 4-{[(4-methylpyridin-3-yl)methyl][6-(trifluoromethyl)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000309_0001
In a reaction vial equipped with a stir bar, N-[(4-methylpyridin-3-yl)methyl]-6-(trifluoromethyl)-1,3- benzothiazol-2-amine (250 mg, 0.77 mmol) was dissolved in dichloromethane (4 mL). To the reaction solution was added 1-(methoxycarbonyl)piperidine-4-carboxylic acid (288 mg, 1.5 mmol) and 50% propylphosphonic anhydride in ethyl acetate (1.45 mL, 1.9 mmol). The reaction mixture was cooled to 0 °C and triethylamine (0.54 mL, 3.9 mmol) was added. The reaction was stirred from 0 °C to room temperature overnight. The product was indicated present via UPLC analysis. The reaction solution was diluted with dichloromethane and washed with water (3 x 15 mL). The organic layers were pooled, dried over anhydrous sodium sulfate, filtered and concentrated. The product was purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane. Product methyl 4-{[(4- methylpyridin-3-yl)methyl][6-(trifluoromethyl)-1,3-benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate (57.0 mg, 0.12 mmol, 15 %) was afforded as a pale yellow solid.1H NMR (300 MHz, Chloroform-d) δ 8.45 (d, J = 4.9 Hz, 1H), 8.25 – 8.02 (m, 2H), 7.80 (d, J = 8.5 Hz, 1H), 7.65 (d, J = 8.9 Hz, 1H), 7.35 – 7.16 (m, 1H), 5.61 (s, 2H), 4.21 (s, 2H), 3.71 (d, J = 0.8 Hz, 3H), 2.77 (t, J = 12.2 Hz, 2H), 2.53 (s, 3H), 1.89 (dd, J = 30.4, 18.2 Hz, 4H), 1.38 – 1.18 (m, 1H); LCMS (ESI) m/z: 493.1 [M+H]+. Example 261. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-1-[2-(1-methylcyclopropyl)acetyl]-N- [(pyridin-3-yl)methyl]piperidine-4-carboxamide
Figure imgf000309_0002
Step 1: Preparation of 1-[2-(1-methylcyclopropyl)acetyl]piperidine-4-carboxylic acid
Figure imgf000309_0003
In a reaction vial equipped with a stir bar, methyl piperidine-4-carboxylate (400 mg, 2.8 mmol) was dissolved in dichloromethane (11 mL). To the reaction solution was added 2-(1- methylcyclopropyl)acetic acid (318 mg, 2.8 mmol) and 50% propylphosphonic anhydride in ethyl acetate (5.29 mL, 6.97 mmol). The reaction mixture was cooled to 0 °C and triethylamine (1.92 mL, 14 mmol) was added. The reaction was stirred from 0 °C to room temperature overnight. The reaction solution was diluted with dichloromethane and washed with water (3 x 15 mL). The organic layers were pooled, dried over anhydrous sodium sulfate, filtered and concentrated. Crude product purified via flash column chromatography eluting with 0-100% ethyl acetate in dichloromethane. The elutant was pooled and concentrated under reduced pressure to afford an orange oil. The oil was dissolved in tetrahydrofuran (10 mL). To the reaction was added water (4 mL) and lithium hydroxide hydrate (66.2 mg, 1.6 mmol), and the reaction was stirred at room temperature overnight. The reaction was acidified with 1N hydrochloric acid. The reaction was then extracted with ethyl acetate (2x 10 mL), and the organic layer was washed with brine. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Crude product was determined sufficiently pure to continue onto next step via NMR analysis. Product 1-[2-(1- methylcyclopropyl)acetyl]piperidine-4-carboxylic acid (193 mg, 0.86 mmol, 54 %) was afforded as a clear oil.1H NMR (300 MHz, Chloroform-d) δ 4.3 - 4.2 (M, 1H), 4.18 - 3.6 (M, 1H), 3.32 – 2.74 (m, 2H), 2.60 (td, J = 10.5, 5.0 Hz, 1H), 2.37 (s, 2H), 2.12 – 1.84 (m, 3H), 1.82 – 1.46 (m, 4H), 1.38 – 0.74 (m, 4H), 0.55 – 0.23 (m, 4H); LCMS data unavailable. Step 2: Preparation of 6-fluoro-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000310_0001
In a reaction vial, a solution of pyridine-3-carbaldehyde (126 mg, 1.2 mmol) and 6-fluoro-1,3- benzothiazol-2-amine (200 mg, 1.2 mmol) in dry toluene (12 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (102 mg, 2.7 mmol) in dry methanol (12 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 6-fluoro-N-[(pyridin-3- yl)methyl]-1,3-benzothiazol-2-amine (274 mg, 1.1 mmol, 90 %) was afforded as a white solid.1H NMR (300 MHz, Methanol-d4) δ 8.59 (s, 1H), 8.51 – 8.39 (m, 1H), 7.89 (d, J = 8.0 Hz, 1H), 7.54 – 7.27 (m, 3H), 7.01 (tdd, J = 9.0, 2.8, 1.3 Hz, 1H), 4.68 (s, 2H); LCMS (ESI) m/z: 260.0 [M+H]+. Step 3: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-1-[2-(1-methylcyclopropyl)acetyl]-N-[(pyridin-3- yl)methyl]piperidine-4-carboxamide
Figure imgf000310_0002
In a reaction vial equipped with a stir bar, 6-fluoro-N-[(pyridin-3-yl)methyl]-1,3-benzothiazol-2- amine (221 mg, 0.86 mmol) was dissolved in dichloromethane (4.5 mL). To the reaction solution was added 1-[2-(1-methylcyclopropyl)acetyl]piperidine-4-carboxylic acid (193 mg, 0.86 mmol) and 50% propylphosphonic anhydride in ethyl acetate (1.61 mL, 2.1 mmol). The reaction mixture was cooled to 0 °C and triethylamine (0.60 mL, 4.3 mmol) was added. The reaction was stirred from 0 °C to room temperature overnight. The product was indicated present via UPLC analysis. The reaction solution was diluted with dichloromethane and washed with water (3 x 15 mL). The organic layers were pooled, dried over anhydrous sodium sulfate, filtered and concentrated. Crude product was purified via flash column chromatography eluting with 0-10% methanol in dichloromethane. Product N-(6-fluoro-1,3-benzothiazol-2- yl)-1-[2-(1-methylcyclopropyl)acetyl]-N-[(pyridin-3-yl)methyl]piperidine-4-carboxamide (108 mg, 0.23 mmol, 27 %) was afforded as a yellow oil.1H NMR (300 MHz, Chloroform-d) δ 8.70 – 8.50 (m, 2H), 7.83 – 7.45 (m, 3H), 7.38 – 7.12 (m, 2H), 5.95 – 5.40 (m, 1H), 5.31 (d, J = 1.0 Hz, 2H), 4.68 (d, J = 13.4 Hz, 1H), 3.94 (d, J = 14.0 Hz, 1H), 3.17 – 2.82 (m, 2H), 2.61 (s, 1H), 2.34 (s, 1H), 1.97 – 1.55 (m, 4H), 1.15 (s, 3H), 0.41 (dd, J = 8.8, 2.2 Hz, 3H); LCMS (ESI) m/z: 467.6 [M+H]+. Example 262. Preparation of N-[(1,3-thiazol-5-yl)methyl]-N-[6-(trifluoromethyl)-1,3-benzothiazol-2- yl]oxane-4-carboxamide
Figure imgf000311_0001
Step 1: Preparation of N-(thiazol-5-ylmethyl)-6-(trifluoromethyl)-1,3-benzothiazol-2-amine
Figure imgf000311_0002
To a solution of thiazole-5-carbaldehyde (2.18 g, 19 mmol) and 6-(trifluoromethyl)-1,3- benzothiazol-2-amine (3.50 g, 16 mmol) in tetrahydrofuran (50 mL) was added titanium isopropoxide (11.4 g, 40 mmol) in a flamed dried flask. The mixture was heated to 60 °C with stirring for 2 hours. Then, the mixture was cooled to 0 °C and sodium borohydride (0.910 g, 24 mmol) was added. The reaction was warmed to room temperature over 2 hours. The product was indicated via UPLC analysis. The reaction was quenched with 10 mL water, and the organics were extracted with ethyl acetate (5x 30 mL). The organic layers were pooled, washed with brine (3x 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by flash column chromatography through 80 grams of silica gel using a gradient of 0-30 % ethyl acetate in petroleum ether. Product N- (thiazol-5-ylmethyl)-6-(trifluoromethyl)-1,3-benzothiazol-2-amine (1.43 g, 4.5 mmol, 28 %) was obtained as a white solid.1H NMR (400 MHz, Chloroform-d) δ = 8.76 (s, 1H), 7.88 (d, J = 6.8 Hz, 2H), 7.65 - 7.52 (m, 2H), 6.29 (br s, 1H), 4.94 (s, 2H); LCMS (ESI) m/z: 316.0 [M+H]+. Step 2: Preparation of N-[(1,3-thiazol-5-yl)methyl]-N-[6-(trifluoromethyl)-1,3-benzothiazol-2-yl]oxane-4- carboxamide
Figure imgf000311_0003
In a reaction vial, N-[(1,3-thiazol-5-yl)methyl]-6-(trifluoromethyl)-1,3-benzothiazol-2-amine (150 mg, 0.48 mmol) and N,N-diisopropylethylamine (0.25 mL, 1.4 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. oxane-4-carbonyl chloride (210 mg, 1.4 mmol) was added dropwise with stirring and the reaction was stirred for 3 hours. The product was confirmed present via UPLC analysis. 10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-[(1,3-thiazol-5-yl)methyl]-N-[6-(trifluoromethyl)-1,3-benzothiazol-2-yl]oxane-4- carboxamide (171 mg, 0.40 mmol, 84 %) was afforded as a white solid.1H NMR (300 MHz, Acetone-d6) δ 8.92 (d, J = 0.8 Hz, 1H), 8.42 (dd, J = 1.8, 0.9 Hz, 1H), 8.14 (d, J = 0.9 Hz, 1H), 8.04 (dt, J = 8.4, 0.8 Hz, 1H), 7.80 (dd, J = 8.4, 1.8 Hz, 1H), 6.00 (d, J = 1.0 Hz, 2H), 4.04 – 3.85 (m, 2H), 3.56 (td, J = 11.7, 2.7 Hz, 3H), 1.95 – 1.68 (m, 4H); LCMS (ESI) m/z: 428.1 [M+H]+. Example 263. Preparation of N-[(4-methylpyridin-3-yl)methyl]-N-[6-(trifluoromethoxy)-1,3- benzothiazol-2-yl]oxane-4-carboxamide
Figure imgf000312_0001
Step 1: Preparation of N-[(4-methyl-3-pyridyl)methyl]-6-(trifluoromethyl)-1,3-benzothiazol-2-amine
Figure imgf000312_0002
To a solution of 4-methylpyridine-3-carbaldehyde (2.20 g, 18 mmol) and 6-(trifluoromethoxy)-1,3- benzothiazol-2-amine (3.55 g, 15 mmol) in tetrahydrofuran (50 mL) was added titanium isopropoxide (10.8 g, 38 mmol) in a flamed dried flask. The mixture was heated to 60 °C with stirring for 2 hours. Then, the mixture was cooled to 0 °C and sodium borohydride (0.862 g, 23 mmol) was added. The reaction was warmed to room temperature over 2 hours. The product was indicated via UPLC analysis. The reaction was quenched with 10 mL water, and the organics were extracted with ethyl acetate (5x 30 mL). The organic layers were pooled, washed with brine (3x 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by flash column chromatography through 80 grams of silica gel using a gradient of 0-30 % ethyl acetate in petroleum ether. Product N-[(4- methyl-3-pyridyl)methyl]-6-(trifluoromethyl)-1,3-benzothiazol-2-amine (2.32 g, 6.8 mmol, 45 %) was obtained as a white solid.1H NMR (400 MHz, Chloroform-d) δ 8.52 (s, 1H), 8.44 (d, J = 5.0 Hz, 1H), 7.54 - 7.45 (m, 2H), 7.21 - 7.13 (m, 2H), 5.68 (br s, 1H), 4.69 (s, 2H), 2.42 (s, 3H); LCMS (ESI) m/z: 340.0 [M+H]+. Step 2: Preparation of N-[(4-methylpyridin-3-yl)methyl]-N-[6-(trifluoromethoxy)-1,3-benzothiazol-2- yl]oxane-4-carboxamide
Figure imgf000312_0003
In a reaction vial, N-[(4-methylpyridin-3-yl)methyl]-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (150 mg, 0.44 mmol) and N,N-diisopropylethylamine (0.23 mL, 1.3 mmol) were dissolved in dimethylformamide (1 mL) and cooled to 0 °C. oxane-4-carbonyl chloride (196 mg, 1.3 mmol) was added dropwise with stirring and the reaction was stirred for 3 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-[(4-methylpyridin-3-yl)methyl]-N-[6-(trifluoromethoxy)-1,3- benzothiazol-2-yl]oxane-4-carboxamide (135 mg, 0.30 mmol, 68 %) was afforded as a white solid.1H NMR (300 MHz, Acetone-d6) δ 8.34 (d, J = 4.9 Hz, 1H), 8.07 (s, 1H), 8.01 (dd, J = 2.3, 1.1 Hz, 1H), 7.76 (d, J = 8.8 Hz, 1H), 7.37 (ddt, J = 8.8, 1.6, 0.9 Hz, 1H), 7.25 (dt, J = 4.9, 0.8 Hz, 1H), 5.79 (s, 2H), 3.90 (ddd, J = 11.6, 4.6, 1.9 Hz, 2H), 3.37 (td, J = 11.8, 2.2 Hz, 2H), 3.21 (dt, J = 11.2, 3.9 Hz, 1H), 2.52 (s, 3H), 2.01 – 1.79 (m, 2H), 1.79 – 1.61 (m, 2H); LCMS (ESI) m/z: 452.1 [M+H]+. Example 264. Preparation of methyl 4-{[(pyrimidin-5-yl)methyl][6-(trifluoromethoxy)-1,3- benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000313_0001
Step 1: Preparation of N-(pyrimidin-5-ylmethyl)-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine
Figure imgf000313_0002
To a solution of pyrimidine-5-carbaldehyde (1.94 g, 18 mmol) and 6-(trifluoromethyl)-1,3- benzothiazol-2-amine (3.51 g, 15 mmol) in tetrahydrofuran (50 mL) was added titanium isopropoxide (10.6 g, 38 mmol) in a flamed dried flask. The mixture was heated to 60 °C with stirring for 2 hours. Then, the mixture was cooled to 0 °C and sodium borohydride (0.851 g, 23 mmol) was added. The reaction was warmed to room temperature over 2 hours. The product was indicated via UPLC analysis. The reaction was quenched with 10 mL water, and the organics were extracted with ethyl acetate (5x 30 mL). The organic layers were pooled, washed with brine (3x 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by flash column chromatography through 80 grams of silica gel using a gradient of 0-30 % ethyl acetate in petroleum ether. Product N- (pyrimidin-5-ylmethyl)-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (1.76 g, 5.4 mmol, 36 %) was obtained as a white solid.1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 9.10 (s, 1H), 8.84(s, 2H), 8.75-8.72 (m, 1H), 7.82 (d, 1H), 7.47 (d, J = 8.8 Hz, 1H), 7.21-7.19 (dd, J = 1.6, 8.8 Hz, 1H), 4.65 (d, J = 5.6, 2H); LCMS (ESI) m/z: 327.0 [M+H]+. Step 2: Preparation of methyl 4-{[(pyrimidin-5-yl)methyl][6-(trifluoromethoxy)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000313_0003
In a reaction vial, N-[(pyrimidin-5-yl)methyl]-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (200 mg, 0.61 mmol), 1-(methoxycarbonyl)piperidine-4-carboxylic acid (228 mg, 1.2 mmol), and triethylamine (0.43 mL, 3.1 mmol) were dissolved in dichloromethane (1.5 mL) with stirring. The reaction was cooled to 0 °C, and 50% propylphosphonic anhydride in ethyl acetate (1.2 mL, 1.5 mmol) was added. The reaction was warmed to room temperature and stirred overnight. The reaction was diluted with 10 mL deionized water and washed with 2x 10 mL portions of dichloromethane. The organic layers were separated, pooled, and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product methyl 4-{[(pyrimidin-5-yl)methyl][6-(trifluoromethoxy)-1,3-benzothiazol-2-yl]carbamoyl}piperidine- 1-carboxylate (217 mg, 0.44 mmol, 72 %) was afforded as a white solid.1H NMR (300 MHz, Acetone-d6) δ 9.07 (s, 1H), 8.84 (d, J = 0.8 Hz, 2H), 8.10 – 7.91 (m, 1H), 7.83 (dd, J = 8.8, 0.6 Hz, 1H), 7.59 – 7.18 (m, 1H), 5.82 (s, 2H), 4.13 (d, J = 13.5 Hz, 2H), 3.64 (s, 3H), 3.42 (tt, J = 11.2, 3.6 Hz, 1H), 2.93 (s, 2H), 1.93 (d, J = 13.2 Hz, 2H), 1.87 – 1.65 (m, 2H); LCMS (ESI) m/z: 496.2 [M+H]+. Example 265. Preparation of methyl 4-{[(1,3-thiazol-5-yl)methyl][6-(trifluoromethoxy)-1,3- benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000314_0001
Step 1: Preparation of N-(thiazol-5-ylmethyl)-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine
Figure imgf000314_0002
To a solution of thiazole-5-carbaldehyde (3.00 g, 27 mmol) and 6-(trifluoromethoxy)-1,3- benzothiazol-2-amine (4.00 g, 17 mmol) in dry diglyme (50 mL) was added titanium isopropoxide (9.71 g, 34 mmol) in a flamed dried flask. The mixture was heated to 60 °C with stirring for 1.5 hours. Then, the mixture was cooled to 0 °C and sodium borohydride (0.969 g, 26 mmol) was added. The reaction was warmed to room temperature over 2 hours. The product was indicated via UPLC analysis. The reaction was quenched with 10 mL water, and the organics were extracted with ethyl acetate (5x 30 mL). The organic layers were pooled, washed with brine (3x 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by reversed-phase HPLC (30% methanol in water solution condition) to give N-(thiazol-5-ylmethyl)-6-(trifluoromethoxy)-1,3-benzothiazol- 2-amine (2.41 g, 7.1 mmol, 42 %) as a white solid.1H NMR (400 MHz, Chloroform-d) δ 8.75 (s, 1H), 7.87 (s, 1H), 7.52 (d, J=8.8 Hz, 1H), 7.46 (d, J=1.1 Hz, 1H), 7.18 (dd, J=1.4, 8.7 Hz, 1H), 5.70 (br s, 1H), 5.01 (s, 2H); LCMS (ESI) m/z: 332.0 [M+H]+. Step 2: Preparation of methyl 4-{[(1,3-thiazol-5-yl)methyl][6-(trifluoromethoxy)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000314_0003
In a reaction vial, N-[(1,3-thiazol-5-yl)methyl]-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (200 mg, 0.60 mmol), 1-(methoxycarbonyl)piperidine-4-carboxylic acid (224 mg, 1.2 mmol), and triethylamine (0.42 mL, 3.0 mmol) were dissolved in dichloromethane (1.5 mL) with stirring. The reaction was cooled to 0 °C, and 50% propylphosphonic anhydride in ethyl acetate (1.1 mL, 1.5 mmol) was added. The reaction was warmed to room temperature and stirred for 2.5 days. The reaction was diluted with 10 mL deionized water and washed with 2x 10 mL portions of dichloromethane. The organic layers were separated, pooled, and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-75% ethyl acetate in dichloromethane. Product methyl 4-{[(1,3-thiazol-5-yl)methyl][6-(trifluoromethoxy)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate (136 mg, 0.27 mmol, 45 %) was afforded as a yellow solid.1H NMR (300 MHz, Acetone-d6) δ 8.92 (d, J = 0.8 Hz, 1H), 8.28 – 8.09 (m, 1H), 8.03 (d, J = 2.3 Hz, 1H), 7.95 (dt, J = 8.9, 0.6 Hz, 1H), 7.46 (ddt, J = 8.8, 2.6, 0.9 Hz, 1H), 5.97 (s, 2H), 4.15 (d, J = 13.5 Hz, 3H), 3.64 (d, J = 0.9 Hz, 3H), 3.53 (ddt, J = 11.2, 7.6, 3.9 Hz, 1H), 3.02 (s, 3H), 1.86 (d, J = 12.9 Hz, 2H), 1.71 (qd, J = 12.3, 4.2 Hz, 2H); LCMS (ESI) m/z: 501.1 [M+H]+. Example 266. Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(4-methylpyridin-3- yl)methyl]oxane-4-carboxamide
Figure imgf000315_0001
Step 1: Preparation of 6-chloro-N-[(4-methyl-3-pyridyl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000315_0002
To a solution of 4-methylpyridine-3-carbaldehyde (2.83 g, 23 mmol) and 6-chloro-1,3- benzothiazol-2-amine (3.60 g, 20 mmol) in tetrahydrofuran (50 mL) was added titanium isopropoxide (13.8 g, 49 mmol) in a flamed dried flask. The mixture was heated to 60 °C with stirring for 2 hours. Then, the mixture was cooled to 0 °C and sodium borohydride (1.10 g, 29 mmol) was added. The reaction was warmed to room temperature over 2 hours. The product was indicated via UPLC analysis. The reaction was quenched with 10 mL water, and the organics were extracted with ethyl acetate (5x 30 mL). The organic layers were pooled, washed with brine (3x 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by flash column chromatography through 80 grams of silica gel using a gradient of 0-30 % ethyl acetate in petroleum ether. Product 6- chloro-N-[(4-methyl-3-pyridyl)methyl]-1,3-benzothiazol-2-amine (1.74 g, 6.0 mmol, 31 %) was obtained as a white solid.1H NMR (400 MHz, Chloroform-d) δ 8.50 (s, 1H), 8.42 (d, J = 4.8 Hz, 1H), 7.54 (s, 1H), 7.42 (d, J = 8.6 Hz, 1H), 7.25 (br d, J = 0.9 Hz, 1H), 7.13 (d, J = 4.8 Hz, 1H), 5.96 (br s, 1H), 4.67 (s, 2H), 2.40 (s, 3H); LCMS (ESI) m/z: 290.0 [M+H]+. Step 2: Preparation of N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(4-methylpyridin-3-yl)methyl]oxane-4- carboxamide
Figure imgf000316_0001
In a reaction vial, 6-chloro-N-[(4-methylpyridin-3-yl)methyl]-1,3-benzothiazol-2-amine (300 mg, 1.0 mmol) and N,N-diisopropylethylamine (0.54 mL, 3.1 mmol) were dissolved in dimethylformamide (2.5 mL) and cooled to 0 °C. oxane-4-carbonyl chloride (459 mg, 3.1 mmol) was added dropwise with stirring and the reaction was stirred for 3 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-(6-chloro-1,3-benzothiazol-2-yl)-N-[(4-methylpyridin-3-yl)methyl]oxane-4-carboxamide (176 mg, 0.44 mmol, 43 %) was afforded as a white solid.1H NMR (300 MHz, Acetone-d6) δ 8.34 (d, J = 4.9 Hz, 1H), 8.13 – 7.99 (m, 2H), 7.66 (d, J = 8.7 Hz, 1H), 7.40 (dd, J = 8.7, 2.2 Hz, 1H), 7.25 (dt, J = 4.9, 0.8 Hz, 1H), 5.77 (s, 2H), 3.90 (ddd, J = 11.6, 4.4, 1.9 Hz, 2H), 3.36 (td, J = 11.8, 2.3 Hz, 2H), 3.22 (tt, J = 11.2, 3.9 Hz, 1H), 2.52 (s, 3H), 2.00 – 1.79 (m, 2H), 1.77 – 1.62 (m, 2H); LCMS (ESI) m/z: 402.1 [M+H]+. Example 267. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]oxane- 4-carboxamide
Figure imgf000316_0002
Step 1: Preparation of 6-fluoro-N-[(1-tritylimidazol-4-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000316_0003
To a solution of 6-fluoro-1,3-benzothiazol-2-amine (4.00 g, 24 mmol) and 1-tritylimidazole-4- carbaldehyde (8.05 g, 24 mmol) in tetrahydrofuran (60 mL) was added titanium isopropoxide (20.3 g, 71 mmol) in a flamed dried flask. The mixture was stirred at 60 °C for 2 hours. Then, sodium cyanoborohydride (2.24 g, 36 mmol) was added at 0 °C and the mixture was warmed to room temperature over 2 hours. The product was indicated present via UPLC analysis. The mixture was cooled to 0 °C and quenched by the slow addition of saturated ammonium chloride (20 mL). The reaction mixture was filtered over Celite and washed with 3x 100 mL portions of ethyl acetate. The filtrate was collected and concentrated under reduced pressure. Crude product was purified via flash column chromatography through 80 grams of silica gel using a gradient of 0-100% ethyl acetate in petroleum ether. Product 6- fluoro-N-[(1-tritylimidazol-4-yl)methyl]-1,3-benzothiazol-2-amine (4.09 g, 7.7 mmol, 32 %) was obtained as a white solid.1H NMR (400 MHz, Chloroform-d) δ 7.47 - 7.40 (m, 2H), 7.37 - 7.28 (m, 10H), 7.16 - 7.09 (m, 6H), 7.00 (dt, J = 2.5, 9.0 Hz, 1H), 6.84 (s, 1H), 6.15 (br s, 1H), 4.53 (s, 2H); LCMS (ESI) m/z: 491.2 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]oxane-4- carboxamide
Figure imgf000317_0001
In a reaction vial, 6-fluoro-N-{[1-(triphenylmethyl)-1H-imidazol-4-yl]methyl}-1,3-benzothiazol-2- amine (300 mg, 0.61 mmol) and N,N-diisopropylethylamine (318 mL, 1.8 mmol) were dissolved in dimethylformamide (1.5 mL) and cooled to 0 °C. oxane-4-carbonyl chloride (271 mg, 1.8 mmol) was added dropwise with stirring and the reaction was stirred for 3 hours. The product was confirmed present via UPLC analysis.10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-4-yl)methyl]oxane-4- carboxamide (27.6 mg, 0.077 mmol, 13 %) was afforded as a white solid.1H NMR (300 MHz, Acetone-d6) δ 7.80 (dd, J = 8.9, 4.8 Hz, 1H), 7.71 (dd, J = 8.5, 2.6 Hz, 1H), 7.58 (s, 1H), 7.30 – 7.10 (m, 2H), 5.57 (s, 2H), 4.04 – 3.87 (m, 2H), 3.68 – 3.56 (m, 1H), 3.56 – 3.29 (m, 2H), 1.93 – 1.65 (m, 4H); LCMS (ESI) m/z: 361.1 [M+H]+. Example 268. Preparation of methyl 4-{[(4-methylpyridin-3-yl)methyl][6-(trifluoromethoxy)-1,3- benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000317_0002
Step 1: Preparation of N-[(4-methyl-3-pyridyl)methyl]-6-(trifluoromethyl)-1,3-benzothiazol-2-amine
Figure imgf000317_0003
To a solution of 4-methylpyridine-3-carbaldehyde (2.20 g, 18 mmol) and 6-(trifluoromethoxy)-1,3- benzothiazol-2-amine (3.55 g, 15 mmol) in tetrahydrofuran (50 mL) was added titanium isopropoxide (10.8 g, 38 mmol) in a flamed dried flask. The mixture was heated to 60 °C with stirring for 2 hours. Then, the mixture was cooled to 0 °C and sodium borohydride (0.862 g, 23 mmol) was added. The reaction was warmed to room temperature over 2 hours. The product was indicated via UPLC analysis. The reaction was quenched with 10 mL water, and the organics were extracted with ethyl acetate (5x 30 mL). The organic layers were pooled, washed with brine (3x 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Crude product was purified by flash column chromatography through 80 grams of silica gel using a gradient of 0-30 % ethyl acetate in petroleum ether. Product N-[(4- methyl-3-pyridyl)methyl]-6-(trifluoromethyl)-1,3-benzothiazol-2-amine (2.32 g, 6.8 mmol, 45 %) was obtained as a white solid.1H NMR (400 MHz, Chloroform-d) δ 8.52 (s, 1H), 8.44 (d, J = 5.0 Hz, 1H), 7.54 - 7.45 (m, 2H), 7.21 - 7.13 (m, 2H), 5.68 (br s, 1H), 4.69 (s, 2H), 2.42 (s, 3H); LCMS (ESI) m/z: 340.0 [M+H]+. Step 2: Preparation of methyl 4-{[(4-methylpyridin-3-yl)methyl][6-(trifluoromethoxy)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000318_0001
In a reaction vial, N-[(4-methylpyridin-3-yl)methyl]-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (200 mg, 0.59 mmol), 1-(methoxycarbonyl)piperidine-4-carboxylic acid (219 mg, 1.2 mmol), and triethylamine (0.41 mL, 2.9 mmol) were dissolved in dichloromethane (1.5 mL) with stirring. The reaction was cooled to 0 °C, and 50% propylphosphonic anhydride in ethyl acetate (1.1 mL, 1.5 mmol) was added. The reaction was warmed to room temperature and stirred for 2.5 days. The reaction was diluted with 10 mL deionized water and washed with 2x 10 mL portions of dichloromethane. The organic layers were separated, pooled, and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product methyl 4-{[(4-methylpyridin-3-yl)methyl][6-(trifluoromethoxy)-1,3-benzothiazol- 2-yl]carbamoyl}piperidine-1-carboxylate (27.9 mg, 0.055 mmol, 9.33 %) was afforded as a cream solid.1H NMR (300 MHz, Acetone-d6) δ 8.22 (d, J = 4.9 Hz, 1H), 7.96 (s, 1H), 7.89 (dd, J = 2.4, 1.2 Hz, 1H), 7.64 (d, J = 8.8 Hz, 1H), 7.33 – 7.21 (m, 1H), 7.13 (dt, J = 4.8, 0.8 Hz, 1H), 5.67 (s, 2H), 3.97 (d, J = 20.4 Hz, 2H), 3.50 (d, J = 1.0 Hz, 3H), 3.08 (tt, J = 11.1, 3.8 Hz, 1H), 2.72 (d, J = 15.5 Hz, 2H), 2.40 (s, 3H), 1.82 – 1.70 (m, 2H), 1.70 – 1.52 (m, 2H); LCMS (ESI) m/z: 509.2 [M+H]+. Example 269. Preparation of 1-(2-cyclopropylacetyl)-N-(5-fluoro-1,3-benzothiazol-2-yl)-N- [(pyrimidin-5-yl)methyl]piperidine-4-carboxamide
Figure imgf000318_0002
Step 1: Preparation of 5-fluoro-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000318_0003
In a reaction vial, a solution of pyrimidine-5-carbaldehyde (192 mg, 1.8 mmol) and 5-fluoro-1,3- benzothiazol-2-amine (300 mg, 1.8 mmol) in dry toluene (18 mL) was refluxed at 120 °C with activated 4 Å molecular sieves under N2 for 24 hours. The reaction mixture was cooled to 60 °C and then poured into a suspension of sodium borohydride (154 mg, 4.1 mmol) in dry ethyl alcohol (18 mL). The resultant reaction mixture was refluxed at 77°C for 30 min and then cooled to room temperature. The product was indicated present via UPLC analysis. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions of ethyl acetate. The product was purified through flash column chromatography on a 12- gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product 5-fluoro-N- [(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine (193 mg, 0.74 mmol, 42 %) was afforded as an orange solid.1H NMR (300 MHz, Methanol-d4) δ 9.07 (d, J = 1.4 Hz, 1H), 8.85 (d, J = 1.3 Hz, 2H), 7.56 (ddd, J = 8.8, 5.4, 1.3 Hz, 1H), 7.14 (ddd, J = 10.1, 2.6, 1.3 Hz, 1H), 7.00 – 6.76 (m, 1H), 4.71 (s, 2H); LCMS (ESI) m/z: 261.0 [M+H]+. Step 2: Preparation of 1-(2-cyclopropylacetyl)piperidine-4-carboxylic acid
Figure imgf000319_0001
In a reaction vial, 2-cyclopropylacetic acid (1.00 g, 10 mmol) was dissolved in dichloromethane (10 mL). thionyl chloride (0.79 mL, 11 mmol) was added and the reaction was heated to 60 °C and stirred for 2 hours. The reaction was cooled to 0 °C and ethyl piperidine-4-carboxylate (1.90 g, 12 mmol) was added. The reaction was warmed to room temperature overnight. The reaction was diluted with 10 mL deionized water and 10 mL dichloromethane. The layers were separated, and the aqueous layer was washed with 2 x 10 mL portions dichloromethane. The organic layers were pooled and concentrated under reduced pressure. The residue was then re-dissolved in tetrahydrofuran (10 mL) and water (1 mL). Lithium hydroxide hydrate (0.835 g, 20 mmol) was added and reaction was stirred at room temperature overnight. The reaction was washed with 1 M hydrochloric acid and 40 mL ethyl acetate. The layers were separated, and the aqueous layer was washed with 2 x 20 mL portions ethyl acetate. The organic layers were pooled and concentrated under reduced pressure. Crude product 1-(2-cyclopropylacetyl)piperidine- 4-carboxylic acid (1.30 g, 6.2 mmol, 62 %) was afforded as a yellow-orange oil.1H NMR (300 MHz, Methanol-d4) δ 4.35 (dd, J = 10.9, 6.7 Hz, 1H), 3.91 (d, J = 13.9 Hz, 1H), 3.80 – 3.66 (m, 1H), 3.26 – 3.13 (m, 1H), 2.92 – 2.78 (m, 1H), 2.70 – 2.52 (m, 1H), 1.96 – 1.89 (m, 1H), 1.72 – 1.42 (m, 2H), 1.04 – 0.92 (m, 1H), 0.54 (dddd, J = 8.3, 5.7, 4.5, 1.1 Hz, 2H), 0.19 (h, J = 4.6, 4.1 Hz, 2H). Step 3: Preparation of 1-(2-cyclopropylacetyl)-N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5- yl)methyl]piperidine-4-carboxamide
Figure imgf000319_0002
In a reaction vial, 5-fluoro-N-[(pyrimidin-5-yl)methyl]-1,3-benzothiazol-2-amine (150 mg, 0.58 mmol), 1-(2-cyclopropylacetyl)piperidine-4-carboxylic acid (242 mg, 1.2 mmol), and triethylamine (0.40 mL, 2.9 mmol) were dissolved in dichloromethane (1.4 mL) with stirring. The reaction was cooled to 0 °C, and 50% propylphosphonic anhydride in ethyl acetate (1.1 mL, 1.4 mmol) was added. The reaction was warmed to room temperature and stirred for 2.5 days. The reaction was diluted with 10 mL deionized water and washed with 2x 10 mL portions of dichloromethane. The organic layers were separated, pooled, and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product 1-(2-cyclopropylacetyl)-N-(5-fluoro-1,3-benzothiazol-2-yl)-N-[(pyrimidin-5-yl)methyl]piperidine-4- carboxamide (15.2 mg, 0.034 mmol, 5.82 %) was afforded as a white solid.1H NMR (300 MHz, Acetone- d6) δ 9.07 (s, 1H), 8.84 (s, 2H), 7.97 (dd, J = 8.8, 5.3 Hz, 1H), 7.48 (dd, J = 9.9, 2.5 Hz, 1H), 7.17 (td, J = 9.0, 2.5 Hz, 1H), 5.82 (d, J = 2.5 Hz, 2H), 4.59 (d, J = 13.3 Hz, 1H), 3.78 (s, 1H), 3.47 (tt, J = 11.2, 3.8 Hz, 1H), 3.19 (t, J = 12.9 Hz, 1H), 2.66 (q, J = 15.0, 13.8 Hz, 1H), 2.30 (dd, J = 6.7, 2.5 Hz, 2H), 1.93 (s, 1H), 1.90 – 1.55 (m, 3H), 1.08 – 0.95 (m, 1H), 0.61 – 0.36 (m, 2H), 0.19 – 0.03 (m, 2H); LCMS (ESI) m/z: 454.2 [M+H]+. Example 270. Preparation of methyl 4-{[(1H-imidazol-5-yl)methyl][6-(trifluoromethoxy)-1,3- benzothiazol-2-yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000320_0001
Step 1: Preparation of N-[(1H-imidazol-5-yl)methyl]-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine
Figure imgf000320_0002
In a reaction vial, 6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (250 mg, 1.1 mmol) and 1H- imidazole-5-carbaldehyde (99.9 mg, 1.0 mmol) were dissolved in dry dioxane (5 mL). titanium(IV) isopropoxide (0.31 mL, 1.1 mmol) was added and the reaction was stirred at 60 °C for 1 hour. The reaction was cooled to room temperature and sodium borohydride (91.9 mg, 2.4 mmol) was added. The reaction mixture was stirred at room temperature for 4 hours. The product was indicated present via UPLC. The reaction was quenched with excess methanol and stirred until bubbling ceased. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions methanol. The filtrate was concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-10% methanol in ethyl acetate. Product N-[(1H- imidazol-5-yl)methyl]-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (86.3 mg, 0.27 mmol, 26 %) was afforded as an orange solid.1H NMR (300 MHz, Methanol-d4) δ 7.67 (d, J = 1.6 Hz, 1H), 7.57 (s, 1H), 7.47 (dd, J = 8.8, 1.3 Hz, 1H), 7.17 (dd, J = 8.6, 2.1 Hz, 1H), 7.10 (s, 1H), 4.59 (s, 2H); LCMS (ESI) m/z: 315.0 [M+H]+. Step 2: Preparation of methyl 4-{[(1H-imidazol-5-yl)methyl][6-(trifluoromethoxy)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000321_0001
In a reaction vial, N-[(1H-imidazol-5-yl)methyl]-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (86.3 mg, 0.27 mmol), 1-(methoxycarbonyl)piperidine-4-carboxylic acid (102 mg, 0.55 mmol), and triethylamine (0.19 mL, 1.4 mmol) were dissolved in dichloromethane (1.5 mL) with stirring. The reaction was cooled to 0 °C, and 50% propylphosphonic anhydride in ethyl acetate (0.52 mL, 0.69 mmol) was added. The reaction was warmed to room temperature and stirred for 2.5 days. The reaction was diluted with 10 mL deionized water and washed with 2x 10 mL portions of dichloromethane. The organic layers were separated, pooled, and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-5% methanol in dichloromethane. Product methyl 4-{[(1H-imidazol-5-yl)methyl][6-(trifluoromethoxy)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate (9.80 mg, 0.020 mmol, 7.42 %) was afforded as a white solid.1H NMR (300 MHz, Acetone-d6) δ 7.96 (s, 1H), 7.89 (d, J = 8.8 Hz, 1H), 7.59 (s, 1H), 7.40 (d, J = 8.8 Hz, 1H), 7.23 (s, 1H), 5.61 (s, 2H), 4.18 (d, J = 9.5 Hz, 2H), 4.04 – 3.78 (m, 1H), 3.65 (s, 2H), 2.96 (s, 3H), 2.02 – 1.87 (m, 2H), 1.74 (ddd, J = 13.6, 8.5, 2.8 Hz, 2H); LCMS (ESI) m/z: 484.1 [M+H]+. Example 271. Preparation of methyl 4-{[(pyridin-3-yl)methyl][6-(trifluoromethoxy)-1,3-benzothiazol- 2-yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000321_0002
Step 1: Preparation of N-[(pyridin-3-yl)methyl]-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine
Figure imgf000321_0003
In a reaction vial, 6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (250 mg, 1.1 mmol) and pyridine- 3-carbaldehyde (111 mg, 1.0 mmol) were dissolved in dry dioxane (7 mL). titanium(IV) isopropoxide (0.31 mL, 1.1 mmol) was added and the reaction was stirred at 60 °C for 1 hour. The reaction was cooled to room temperature and sodium borohydride (91.9 mg, 2.4 mmol) was added. The reaction mixture was stirred at room temperature for 4 hours. The product was indicated present via UPLC. The reaction was quenched with excess methanol and stirred until bubbling ceased. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions methanol. The filtrate was concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-10% methanol in ethyl acetate. Product N-[(pyridin-3-yl)methyl]-6-(trifluoromethoxy)- 1,3-benzothiazol-2-amine (229 mg, 0.71 mmol, 67 %) was afforded as a cream solid.1H NMR (300 MHz, Methanol-d4) δ 8.60 (s, 1H), 8.45 (d, J = 4.9 Hz, 1H), 8.02 – 7.84 (m, 1H), 7.58 (s, 1H), 7.44 (ddd, J = 8.1, 5.3, 3.1 Hz, 2H), 7.17 (d, J = 8.8 Hz, 1H), 4.71 (s, 2H); LCMS (ESI) m/z: 326.0 [M+H]+. Step 2: Preparation of methyl 4-{[(pyridin-3-yl)methyl][6-(trifluoromethoxy)-1,3-benzothiazol-2- yl]carbamoyl}piperidine-1-carboxylate
Figure imgf000322_0001
In a reaction vial, N-[(pyridin-3-yl)methyl]-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (229 mg, 0.70 mmol), 1-(methoxycarbonyl)piperidine-4-carboxylic acid (262 mg, 1.4 mmol), and triethylamine (0.49 mL, 3.5 mmol) were dissolved in dichloromethane (2 mL) with stirring. The reaction was cooled to 0 °C, and 50% propylphosphonic anhydride in ethyl acetate (1.3 mL, 1.8 mmol) was added. The reaction was warmed to room temperature and stirred for 2.5 days. The reaction was diluted with 10 mL deionized water and washed with 2x 10 mL portions of dichloromethane. The organic layers were separated, pooled, and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-100% ethyl acetate in hexanes. Product methyl 4-{[(pyridin-3-yl)methyl][6-(trifluoromethoxy)-1,3-benzothiazol-2-yl]carbamoyl}piperidine-1- carboxylate (53.3 mg, 0.11 mmol, 15 %) was afforded as a yellow solid.1H NMR (300 MHz, Acetone-d6) δ 8.89 – 8.57 (m, 1H), 8.51 (dd, J = 4.8, 1.6 Hz, 1H), 8.01 (dd, J = 2.4, 1.1 Hz, 1H), 7.82 (d, J = 8.8 Hz, 1H), 7.72 (ddd, J = 7.9, 2.5, 1.6 Hz, 1H), 7.47 – 7.18 (m, 2H), 5.84 (s, 2H), 4.10 (d, J = 13.0 Hz, 2H), 3.63 (d, J = 0.8 Hz, 3H), 3.31 (ddt, J = 10.8, 7.8, 4.0 Hz, 1H), 2.88 (s, 2H), 1.93 – 1.56 (m, 4H); LCMS (ESI) m/z: 495.1 [M+H]+. Example 272. Preparation of N-[(1,3-thiazol-2-yl)methyl]-N-[6-(trifluoromethoxy)-1,3-benzothiazol-2- yl]oxane-4-carboxamide
Figure imgf000322_0002
Step 1: Preparation of N-[(1,3-thiazol-2-yl)methyl]-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine
Figure imgf000322_0003
In a reaction vial, 6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (250 mg, 1.1 mmol) and 1,3- thiazole-2-carbaldehyde (117 mg, 1.0 mmol) were dissolved in dry dioxane (7 mL). Titanium(IV) isopropoxide (0.31 mL, 1.1 mmol) was added and the reaction was stirred at 60 °C for 1 hour. The reaction was cooled to room temperature and sodium borohydride (91.9 mg, 2.4 mmol) was added. The reaction mixture was stirred at room temperature for 4 hours. The product was indicated present via UPLC. The reaction was quenched with excess methanol and stirred until bubbling ceased. The reaction was filtered over a bed of Celite and washed with 2x 20 mL portions methanol. The filtrated was concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-10% methanol in ethyl acetate. Product N-[(1,3-thiazol- 2-yl)methyl]-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (280 mg, 0.85 mmol, 80 %) was afforded as an orange solid.1H NMR (300 MHz, Methanol-d4) δ 7.75 (dd, J = 3.4, 1.5 Hz, 1H), 7.61 (s, 1H), 7.52 (dd, J = 3.5, 1.5 Hz, 1H), 7.47 (dd, J = 8.8, 1.5 Hz, 1H), 7.18 (d, J = 8.7 Hz, 1H), 4.98 (d, J = 1.5 Hz, 2H); LCMS (ESI) m/z: 331.9 [M+H]+. Step 2: Preparation of N-[(1,3-thiazol-2-yl)methyl]-N-[6-(trifluoromethoxy)-1,3-benzothiazol-2-yl]oxane-4- carboxamide
Figure imgf000323_0001
In a reaction vial, N-[(1,3-thiazol-2-yl)methyl]-6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (281 mg, 0.85 mmol) and N,N-diisopropylethylamine (0.44 mL, 2.5 mmol) were dissolved in tetrahydrofuran (2 mL) and cooled to 0 °C before oxane-4-carbonyl chloride (750 mg, 5.1 mmol) was added dropwise with stirring and the reaction was stirred for overnight. The product was confirmed present via UPLC analysis. 10 mL of deionized water and 3x 10 mL portions of ethyl acetate were added to the reaction and the organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-75% ethyl acetate in hexanes. Product N-[(1,3-thiazol-2-yl)methyl]-N-[6-(trifluoromethoxy)-1,3-benzothiazol-2-yl]oxane-4- carboxamide (100 mg, 0.23 mmol, 27 %) was afforded as a yellow solid.1H NMR (300 MHz, Acetone-d6) δ 8.01 (dd, J = 2.4, 1.1 Hz, 1H), 7.88 (dd, J = 8.8, 0.8 Hz, 1H), 7.75 (dd, J = 3.3, 0.9 Hz, 1H), 7.59 (dd, J = 3.3, 0.9 Hz, 1H), 7.49 – 7.36 (m, 1H), 6.01 (s, 2H), 4.05 – 3.84 (m, 2H), 3.63 (tt, J = 11.1, 4.2 Hz, 1H), 3.47 (td, J = 11.6, 2.7 Hz, 2H), 1.90 – 1.56 (m, 4H); LCMS (ESI) m/z: 444.1 [M+H]+. Example 273. Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-2- (oxan-4-yl)acetamide
Figure imgf000323_0002
Step 1: Preparation of 6-fluoro-N-[(1-tritylimidazol-4-yl)methyl]-1,3-benzothiazol-2-amine
Figure imgf000323_0003
To a solution of 6-fluoro-1,3-benzothiazol-2-amine (4.00 g, 24 mmol) and 1-tritylimidazole-4- carbaldehyde (8.05 g, 24 mmol) in tetrahydrofuran (60 mL) was added titanium isopropoxide (20.3 g, 71 mmol) in a flamed dried flask. The mixture was stirred at 60 °C for 2 hours. Then, sodium cyanoborohydride (2.24 g, 36 mmol) was added at 0 °C and the mixture was warmed to room temperature over 2 hours. The product was indicated present via UPLC analysis. The mixture was cooled to 0 °C and quenched by the slow addition of saturated ammonium chloride (20 mL). The reaction mixture was filtered over Celite and washed with 3x 100 mL portions of ethyl acetate. The filtrate was collected and concentrated under reduced pressure. Crude product was purified via flash column chromatography through 80 grams of silica gel using a gradient of 0-100% ethyl acetate in petroleum ether. Product 6- fluoro-N-[(1-tritylimidazol-4-yl)methyl]-1,3-benzothiazol-2-amine (4.09 g, 7.7 mmol, 32 %) was obtained as a white solid.1H NMR (400 MHz, Chloroform-d) δ 7.47 - 7.40 (m, 2H), 7.37 - 7.28 (m, 10H), 7.16 - 7.09 (m, 6H), 7.00 (dt, J = 2.5, 9.0 Hz, 1H), 6.84 (s, 1H), 6.15 (br s, 1H), 4.53 (s, 2H); LCMS (ESI) m/z: 491.2 [M+H]+. Step 2: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-2-(oxan-4-yl)-N-{[1-(triphenylmethyl)-1H- imidazol-5-yl]methyl}acetamide
Figure imgf000324_0001
In a reaction vial, 6-fluoro-N-{[1-(triphenylmethyl)-1H-imidazol-5-yl]methyl}-1,3-benzothiazol-2- amine (250 mg, 0.51 mmol), 2-(oxan-4-yl)acetic acid (145 mg, 1.0 mmol), and 50% propylphosphonic anhydride in ethyl acetate (0.96 mL, 1.3 mmol) were dissolved in dichloromethane (1 mL) with stirring. The reaction was cooled to 0 °C, and triethylamine (0.35 mL, 2.5 mmol) was added. The reaction was warmed to room temperature and stirred for 2.5 days. The reaction was diluted with 10 mL deionized water and washed with 2x 10 mL portions of dichloromethane. The organic layers were separated, pooled, and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-25% ethyl acetate in hexanes. Product N-(6-fluoro-1,3-benzothiazol-2-yl)-2-(oxan-4-yl)-N-{[1-(triphenylmethyl)-1H-imidazol-5- yl]methyl}acetamide (136 mg, 0.22 mmol, 43 %) was afforded as a white solid.1H NMR (300 MHz, Chloroform-d) δ 7.44 – 7.34 (m, 1H), 7.29 (dt, J = 8.1, 2.3 Hz, 1H), 7.18 (d, J = 2.4 Hz, 1H), 7.06 (tt, J = 8.4, 5.2 Hz, 9H), 7.00 – 6.82 (m, 8H), 6.75 (s, 1H), 5.17 (s, 2H), 3.91 – 3.64 (m, 2H), 3.29 (td, J = 11.8, 2.2 Hz, 2H), 2.80 (d, J = 6.5 Hz, 2H), 2.12 (ddd, J = 11.2, 7.5, 4.0 Hz, 1H), 1.73 – 1.47 (m, 2H), 1.22 (qd, J = 12.2, 4.0 Hz, 2H). Step 3: Preparation of N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5-yl)methyl]-2-(oxan-4- yl)acetamide
Figure imgf000325_0001
In a reaction vial, N-(6-fluoro-1,3-benzothiazol-2-yl)-2-(oxan-4-yl)-N-{[1-(triphenylmethyl)-1H- imidazol-5-yl]methyl}acetamide (136 mg, 0.22 mmol) was dissolved in dichloromethane (1 mL). trifluoroacetic acid (0.34L, 4.4 mmol) was added and the reaction was stirred overnight. The product was indicated present via UPLC analysis. A solution of saturated sodium bicarbonate was added to the reaction until bubbling ceased. The layers were separated, and the aqueous layer was washed with 2 x 10 mL dichloromethane. The organic layers were pooled and concentrated under reduced pressure. The product was purified through flash column chromatography on a 12-gram silica gel column using a gradient of 0-10% methanol in ethyl acetate. Product N-(6-fluoro-1,3-benzothiazol-2-yl)-N-[(1H-imidazol-5- yl)methyl]-2-(oxan-4-yl)acetamide (26.0 mg, 0.069 mmol, 32 %) was afforded as a white solid.1H NMR (300 MHz, Acetone-d6) δ 7.80 (dd, J = 8.9, 4.8 Hz, 1H), 7.71 (dd, J = 8.5, 2.7 Hz, 1H), 7.58 (s, 1H), 7.27 – 7.13 (m, 2H), 5.49 (s, 2H), 3.94 – 3.79 (m, 2H), 3.69 – 3.55 (m, 2H), 3.39 (td, J = 11.7, 2.1 Hz, 2H), 3.16 (d, J = 7.0 Hz, 2H), 2.35 – 2.14 (m, 1H), 1.46 – 1.24 (m, 3H); LCMS (ESI) m/z: 375.1 [M+H]+. Example 274. Inhibition of CYP51A1 by Compounds of the Invention Method: Recombinant human CYP51A1 (lanosterol-14a-demethylase) enzyme was co-expressed with CYP reductase in bacterial membranes and the fluorescent substrate BOMCC (a non-natural substrate that causes increases in fluorescence upon CYP51A1-dependent demethylation) was used to obtain 8-point dose concentration-response curves for each compound. Results: As shown in Table 4, the compounds of the invention inhibit CYP51A1. Table 4.
Figure imgf000325_0002
Figure imgf000326_0001
“+++” = ≤0.1 µM; “++” = 0.1 µM to ≤1 µM; “+” = >1 µM Example 275. Inhibition of CYP51A1 modulates TDP-43 aggregation Introduction Amyotrophic lateral sclerosis (ALS), also known as Lou Gehrig’s disease, is an aggressive, debilitating disease in which affected patients succumb within two to five years after diagnosis. ALS presents with heterogeneous clinical features but has a common underlying pathology of motor neuron loss that limits the central nervous system’s ability to effectively regulate voluntary and involuntary muscle activity. Additionally, without neuronal trophic support muscles being to atrophy, further exacerbating motor deterioration. Cellular and tissue degeneration results in motor impairment such as fasciculations and weakening in the arms, legs and neck, difficulty swallowing, slurred speech and ultimately failure of the diaphragm muscles that control breathing. At the cellular level, 97% of all ALS cases have the common pathological feature of misfolded and aggregated TAR-DNA binding protein (TDP)-43 in spinal motor neuron inclusions. TDP-43 is a DNA/RNA binding protein involved in RNA splicing and is typically localized to the nucleus but can be translocated to the cytoplasm under conditions of cell stress. Nuclear clearing and cytoplasmic accumulation of misfolded and aggregated TDP-43 are hallmarks of degenerating motor neurons in ALS, but it remains unclear if mechanism of toxicity is due to aggregation-dependent loss of TDP-43 function or if the aggregates acquire toxic gain of function. Aggregates of TDP-43 accumulate in discrete cellular domains known as stress granules, which are also enriched with translationally inactive mRNAs. Stress granules are observed in multiple cellular types and are thought to be directly related to TDP-43- dependent toxicity in ALS and FTD. Dysfunction in DNA/RNA binding protein activity plays a crucial role in susceptible motor neurons in ALS, as familial cases have also been traced to mutations in the protein Fused in Sarcoma (FUS), a DNA/RNA binding protein that recently has been shown to be involved in gene silencing. Preclinical studies suggest that FUS mutations promote a toxic gain of function that may be causative in motor neuron degeneration. Mutations in the TDP-43 gene (TARDBP) have also been causally linked to familial forms of ALS. A common TDP-43 mutation is known as Q331K, in which glutamine (Q) 331 has been mutated to a lysine (K). This mutation results in a TDP-43 protein that is more aggregation prone and exhibits enhanced toxicity. A recent study has also demonstrated that the Q331K mutation can confer a toxic gain of function in a TDP-43 knock-in mouse, which exhibits cognitive deficits and histological abnormalities similar to that which occurs in frontotemporal dementia (FTD). FTD refers to a group of degenerative disorders that are characterized by atrophy in the frontal and temporal cortices due to progressive neuron loss. Due to the functional nature of the brain regions impacted in FTD, the most common symptoms involve noticeable alterations in personality, behavior and linguistic ability and can also present with loss of speech. The pathological basis of FTD appears to be multifactorial involving mutations in genes such as C9orf72, progranulin (GRN) and MAPT, but intracellular inclusions of aggregated TDP-43, FUS and tau have been observed. Although ALS and FTD may have different genetic and molecular triggers and occur in different cell types, similar protein misfolding and degenerative mechanisms may operate in multiple diseases. The toxic gain of function features of TDP-43 can be faithfully recapitulated in the simple model organism, budding yeast, where the protein also localizes to stress granules. Human disease mutations in TDP-43 enhance toxicity and yeast genetic screens have revealed key connections that are conserved to humans. The yeast model thus provides a robust cell-based screening platform for small molecules capable of ameliorating toxicity. To validate compounds from such phenotypic screens, it is imperative to test compounds in a mammalian neuronal context. In an effort to develop TDP-43-related mammalian models of neuron loss that occurs in ALS and FTD, primary cultures of rat cortical neurons were transfected with human wild type or Q331K mutant TDP-43. These cells were compared to cells which received an empty expression vector control. Validation studies have demonstrated that cells expressing either wild type or Q331K TDP-43 have are more susceptible to dying over time in culture. In the experiments described in this example, this model system is used to interrogate new therapeutic approaches to ameliorate TDP-43 toxicity. Results From the TDP-43 yeast model, a compound with known mode of action was identified that restored viability to TDP-43-expressing yeast (FIG.1A). Fluconazole is an antifungal known to inhibit Erg11, the yeast lanosterol 14-alpha demethylase (FIG.1B). Inhibition of Erg11 reduces ergosterol synthesis (yeast equivalent of cholesterol), while increasing lanosterol levels, the substrate of Erg11 (FIG. 1C). The human homolog of Erg11 is Cyp51A1, a member of the cytochrome P450 superfamily of enzymes but does not appear to have a role in detoxification of xenobiotics. CYP51A1 has also been known as lanosterol 14-alpha demethylase, which describes its function in removing the 14-alpha-methyl group from lanosterol to generate 4,4-dimethylcholesta-8(9),14,24-trien-3β-ol, which is a critical step in the cholesterol biosynthetic pathway. To evaluate the potential role of CYP51A1 in TDP-43 pathology, the aforementioned primary rat cortical neuron TDP-43 models were utilized to test the efficacy of published inhibitors (FIG.2). Rat cortical neurons transfected with wild type human TDP-43 exhibited a significant reduction in survival compared to neurons transfected with empty vector control, and this reduction in survival was partially alleviated by treatment with compound A (FIGS.3A and 3B). Compound A has the structure:
Figure imgf000328_0001
A similar survival befit was conferred by compound A when applied to cells transfected with Q331K mutant TDP-43 (FIGS.4A and 4B). A similar effect in rescuing a survival deficit was observed for a structurally differentiated compound, compound B, when applied to cells transfected with wild-type TDP-43 (FIGS.5A and 5B). Compound B has the structure:
Figure imgf000328_0002
These studies demonstrate that inhibition of Erg11 in yeast and inhibition of Cyp51A1 has a beneficial effect of rescuing cells from wild type and mutant TDP-43 toxicity and promotes cell survival. This is the first demonstration that inhibition of CYP51A1 is beneficial in treating and preventing TDP-43 pathological processes and represents a novel therapeutic approach for the treatment of ALS. Other Embodiments While the present invention has been described with reference to what are presently considered to be the preferred examples, it is to be understood that the invention is not limited to the disclosed examples. To the contrary, the invention is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. All publications, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety. Where a term in the present application is found to be defined differently in a document incorporated herein by reference, the definition provided herein is to serve as the definition for the term. Other embodiments are in the claims.

Claims

CLAIMS 1. A compound, or a pharmaceutically acceptable salt thereof, having the structure:
Figure imgf000330_0001
Formula I wherein n is 0, 1, 2, 3, or 4; X is S or O; each R1 is, independently, halo, cyano, hydroxy, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, -SO2-optionally substituted C1-C6 alkyl, or -CO2-optionally substituted C1-C6 alkyl; R2 is optionally substituted C2-C9 heteroaryl; and R3 is optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C9 heterocyclyl, optionally substituted C1-C6 alkyl C3-C8 cycloalkyl, optionally substituted C1-C6 heteroalkyl C3-C8 cycloalkyl, optionally substituted C1-C6 alkyl C2-C9 heteroaryl, optionally substituted C1-C6 heteroalkyl C2-C9 heteroaryl, optionally substituted C1-C6 alkyl C2-C9 heterocyclyl, optionally substituted C1-C6 heteroalkyl C2-C9 heterocyclyl, optionally substituted C1-C6 alkyl C6-C10 aryl, or optionally substituted C1-C6 heteroalkyl C6-C10 aryl.
2. The compound of claim 1, wherein X is S.
3. The compound of claim 1, wherein X is O.
4. The compound of any one of claims 1 to 3, wherein n is 0.
5. The compound of any one of claims 1 to 3, wherein n is 1.
6. The compound of claim 5, wherein the compound has the structure:
Figure imgf000330_0002
. Formula Ia
7. The compound of claim 5, wherein the compound has the structure:
Figure imgf000330_0003
. Formula Ib
8. The compound of claim 5, wherein the compound has the structure: .
Figure imgf000331_0001
9. The compound of claim 5, wherein the compound has the structure:
Figure imgf000331_0002
. Formula Id
10. The compound of any one of claims 1 to 9, wherein R1 is halo.
11. The compound of claim 10, wherein halo is fluoro.
12. The compound of any one of claims 1 to 9, wherein R1 is cyano.
13. The compound of any one of claims 1 to 9, wherein R1 is hydroxy.
14. The compound of any one of claims 1 to 9, wherein R1 is optionally substituted C1-C6 alkyl.
15. The compound of claim 14, wherein optionally substituted C1-C6 alkyl is methyl, ethyl, trifluoromethyl, or hydroxymethyl.
16. The compound of any one of claims 1 to 9, wherein R1 is optionally substituted C1-C6 heteroalkyl.
17. The compound of claim 16, wherein optionally substituted C1-C6 heteroalkyl is ethoxy or trifluoromethoxy.
18. The compound of any one of claims 1 to 9, wherein R1 is -SO2-optionally substituted C1-C6 alkyl.
19. The compound of claim 18, wherein -SO2-optionally substituted C1-C6 alkyl is -SO2-methyl.
20. The compound of any one of claims 1 to 9, wherein R1 is -CO2-optionally substituted C1-C6 alkyl.
21. The compound of claim 20, wherein -CO2-optionally substituted C1-C6 alkyl is -CO2-methyl or -CO2-ethyl.
22. The compound of any one of claims 1 to 21, wherein R2 is a 5-membered optionally substituted C2-C9 heteroaryl.
23. The compound of claim 22, wherein R2 is
Figure imgf000332_0001
, , , ,
Figure imgf000332_0002
24. The compound of any one of claims 1 to 21, wherein R2 is a 6-membered optionally substituted C2-C9 heteroaryl. 25. The compound of claim 24, wherein R2 is
Figure imgf000332_0003
, , , ,
Figure imgf000332_0004
26. The compound of any one of claims 1 to 25, wherein R3 is optionally substituted C3-C8 cycloalkyl. 27. The compound of claim 26, wherein R3 is cyclohexyl. 28. The compound of any one of claims 1 to 25, wherein R3 is optionally substituted C2-C9 heterocyclyl. 29. The compound of claim 28, wherein R3 is
Figure imgf000332_0005
, , , ,
Figure imgf000332_0006
, wherein R4 is -C(O)R5; R5 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C1-C6 alkyl C3-C8 cycloalkyl, or optionally substituted C1-C6 heteroalkyl C3-C8 cycloalkyl. 30. The compound of any one of claims 1 to 25, wherein R3 is optionally substituted C6-C10 aryl. 31. The compound of claim 30, wherein R3 is phenyl, 4-cyano-phenyl, or 4-fluoro-phenyl. 32. A compound, or pharmaceutically acceptable salt thereof, having the structure of any one of compounds 1-273 in Table 1. 33. A pharmaceutical composition comprising a compound of any one of claims 1 to 32 and a pharmaceutically acceptable excipient. 34. A method of treating a neurological disorder in a subject in need thereof, the method comprising administering an effective amount of a compound, or pharmaceutically acceptable salt thereof, of any one of claims 1 to 32 or a pharmaceutical composition of claim 33. 35. A method of inhibiting toxicity in a cell related to a protein, the method comprising administering an effective amount of a compound of any one of claims 1 to 32 or a pharmaceutical composition of claim 33. 36. The method of claim 35, wherein the toxicity is TDP-43-related toxicity. 37. The method of any one of claims 35 to 36, wherein the cell is a mammalian neural cell. 38. A method of treating a CYP51A1-associated disorder in a subject in need thereof, the method comprising administering an effective amount of a compound, or pharmaceutically acceptable salt thereof, of any one of claims 1 to 32 or a pharmaceutical composition of claim 33. 39. The method of claim 38, wherein the CYP51A1-associated disorder is ALS. 40. A method of inhibiting CYP51A1, the method comprising contacting a cell with an effective amount of a compound of any one of claims 1 to 32 or a pharmaceutical composition of claim 33.
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Citations (2)

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US20100075959A1 (en) * 2000-06-21 2010-03-25 Alexander Alanine Benzothiazole derivatives with activity as adenosine receptor ligands
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US20190134056A1 (en) * 2017-03-10 2019-05-09 The Trustees Of The Stevens Institute Of Technolog K-ras mutations and antagonists

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Title
DATABASE Pubchem NCBI; 25 January 2012 (2012-01-25), ANONYMOUS : "1-(3-chlorophenyl)-N-[4-[(1-methylimidazol-2-yl)methoxy]phenyl]cyclopentane-1-carboxamide | C23H24ClN3O2", XP055881736, Database accession no. PubChem CID 55881734 *
DATABASE PubChem NCBI; 30 July 2007 (2007-07-30), ANONYMOUS : "[1-(Methylcarbamoylamino)-1-oxopropan-2-yl] 4-phenylmethoxybenzoate | C19H20N2O5", XP055881734, Database accession no. PubChem CID 16503869 *

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