structure–activity relationship studies of tolfenpyrad

1 Structure−Activity Relationship Studies of Tolfenpyrad Reveal2 Subnanomolar Inhibitors of Haemonchus contortus Development3 Thuy G. Le,‡ Abhijit Kundu,§ Atanu Ghoshal,§ Nghi H. Nguyen,‡ Sarah Preston,⊥,# Yaqing Jiao,⊥

4 Banfeng Ruan,‡,∇ Lian Xue,† Fei Huang,† Jennifer Keiser,○,⧫ Andreas Hofmann,∥

5 Bill C. H. Chang,⊥ Jose Garcia-Bustos,⊥ Timothy N. C. Wells,¶ Michael J. Palmer,¶ Abdul Jabbar,⊥

6 Robin B. Gasser,*,⊥ and Jonathan B. Baell*,†,‡

7†School of Pharmaceutical Sciences, Nanjing Tech University, No. 30, South Puzhu Road, Nanjing 211816, People’s Republic of

8 China

9‡Medicinal Chemistry, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Melbourne, Victoria 3052,

10 Australia

11§TCG Lifesciences Private Limited, Block BN, Plot 7, Salt-Lake Electronics Complex, Sector V, Kolkata 700091, West Bengal, India

12∥Griffith Institute for Drug Discovery, Griffith University, Nathan, Brisbane, Queensland4111, Australia

13⊥Department of Veterinary Biosciences, Melbourne Veterinary School, Faculty of Veterinary and Agricultural Sciences, The

14 University of Melbourne, Parkville, Melbourne, Victoria 3010, Australia

15#School of Health and Life Sciences, Federation University, Ballarat, Victoria 3353, Australia

16∇School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, P. R. China

17○Swiss Tropical and Public Health Institute, 4051 Basel, Switzerland

18⧫University of Basel, 4001 Basel, Switzerland

19¶Medicines for Malaria Venture, 1215 Geneva, Switzerland

20 *S Supporting Information

21 ABSTRACT: Recently, we have discovered that the regis-22 tered pesticide, tolfenpyrad, unexpectedly and potently23 inhibits the development of L4 larval stages of the parasitic24 nematode Haemonchus contortus with an IC50 value of 0.0325 μM while displaying good selectivity, with an IC50 of 37.9 μM26 for cytotoxicity. As a promising molecular template for27 medicinal chemistry optimization, we undertook anthelmintic28 structure−activity relationships for this chemical. Modifica-29 tions of the left-hand side, right-hand side, and middle section30 of the scaffold were explored to produce a set of 57 analogues. Analogues 25, 29, and 33 were shown to be the most potent31 compounds of the series, with IC50 values at a subnanomolar level of potency against the chemotherapeutically relevant fourth32 larval (L4) stages of H. contortus. Selected compounds from the series also showed promising activity against a panel of other33 different parasitic nematodes, such as hookworms and whipworms.

34 ■ INTRODUCTION

35 Parasitic worms, particularly gastrointestinal roundworms36 (nematodes), are major pathogens of livestock animals and37 cause diseases that, through productivity losses, adversely38 impact the agricultural, meat, and dairy industries.1,2 The39 control of these worms relies heavily on the use of anthelmintic40 chemotherapy. However, the effectiveness of many anthelmin-41 tics around the world has significantly decreased due to42 widespread drug resistance in such worms resulting from the43 excessive and uncontrolled use of these drugs.3−6 Therefore,44 the discovery of new anthelmintics with novel modes of action45 and that are active against drug-resistant parasites is in high46 demand.7

47 f1Recently, we have identified tolfenpyrad (TFP, Figure 1) to48be potently inhibitory of the motility and development of49parasitic larvae of Haemonchus contortus,8,9 a parasitic50nematode of major economic importance in ruminants. TFP51is a registered pesticide used in many countries to control52arthropod pests on infested crops.10 Along with the closely53related tebufenpyrad, it belongs to the pyrazole-5-carboxamide54class of complex I inhibitors, which interrupt electron transport55through inhibiting NADH/ubiquinone oxoreductase.11 De-56spite being reported in 1996,12 published SAR interrogation of57tolfenpyrad is relatively limited, even though marked

Received: November 15, 2018Published: December 20, 2018

Article

pubs.acs.org/jmc

© XXXX American Chemical Society A DOI: 10.1021/acs.jmedchem.8b01789J. Med. Chem. XXXX, XXX, XXX−XXX

* Unknown * | ACSJCA | JCA11.1.4300/W Library-x64 | research.3f (R4.1.i3 HF01:4938 | 2.1) 2018/08/24 11:08:00 | PROD-WS-120 | rq_779338 | 12/27/2018 09:34:26 | 18 | JCA-DEFAULT

pubs.acs.org/jmc

http://dx.doi.org/10.1021/acs.jmedchem.8b01789

58 insecticidal, fungicidal, or miticidal activities have been59 observed for this chemical.12−15 Furthermore, nothing is60 known about its SAR against parasitic nematodes. Herein, by61 utilizing a well-established but proprietary and sophisticated62 phenotypic drug-screening platform for H. contortus,16,17 we63 report, for the first time, comprehensive SAR of TFP for64 inhibition of L3 motility and L4 development of H. contortus65 larvae and reveal novel modifications that reach subnanomolar66 levels of L4 larval development inhibition.67 It can be seen that TFP harbors a large, hydrophobic,68 electron-rich p-methylphenoxybenzyloxy group. From a69 medicinal chemistry perspective, this chemical property of70 TFP would be considered suboptimal for a drug candidate that71 is proposed to be administered, for example, orally to a72 vertebrate animal affected by worms, as opposed to the73 application (as pesticide) to the surface of arthropod-affected74 plants. Therefore, the predominant focus of this study was to75 explore TFP SAR with a view to maintaining or even76 increasing potency of TFP, while moving the scaffold into a77 more druglike physicochemical “space” that might impart78 improved solubility and metabolic stability properties.

79 ■ RESULTS AND DISCUSSION80 The structure of the TFP scaffold can usefully be considered as81 divided into two main components, namely, the pyrazole-5-82 carboxamide and the p-methylphenoxybenzyloxy parts. For83 late-stage derivatization of the RHS pyrazole, the p-84 methylphenoxybenzyloxy group was obtained via a nucleo-85 philic aromatic substitution reaction of p-cresol and 4-86 fluorobenzonitrile, followed by a reduction of the nitrile87 group, to yield the benzylamine, which was then reacted with88 the pyrazole-5-carboxylic acid via an amide coupling reaction

s1 89 (Scheme 1). Likewise, in some cases, the LHS derivatization

90process could be performed in reverse order so that the RHS91pyrazole was to be installed first to form a 4-halobenzylamide92intermediate, which was then subjected to an Ullmann-type93 s2coupling with a phenoxy species (Scheme 2). The CuI/N,N-94dimethylglycine was an efficient catalytic system for the95Ullmann-type coupling, as reported by Ma et al.18

96To examine SAR, compounds were first subjected to a97primary screen to assess their ability to inhibit the motility of98H. contortus at the exsheathed L3 (xL3) stage, using99monepantel and moxidectin as positive control anthelmintics.100Only compounds that resulted in ≥70% motility inhibition of101xL3 larvae at a concentration of 100 μM were subjected to102subsequent dose−response evaluation, to establish IC50 values,103and then further assessed in the H. contortus L4 development104assay. The first aim of the study was to explore the chemical105space on the RHS pyrazole of TFP and simultaneously to106reduce the overall hydrophobicity by removing some or all of107the substituents on the pyrazole and to introduce functional108groups with differing steric and electronic effects. The results109 t1of this exploration are summarized in Table 1. Loss of both 3-110Et and 4-Cl on the pyrazole ring (3) led to a moderate111decrease in inhibitory potency of xL3 motility and a slight112reduction in L4 development compared to TFP. A similar but113diminished loss of activity was observed when the 3-Et group114was maintained but the 4-Cl group removed, to give 4, which115still exhibited relatively potent L4 larval development116inhibition (IC50 0.057 μM). Hence, comparing 3 with 4117suggests some hydrophobicity on the 3-position, which is118favorable. Implementing nitrile or trifluoromethyl groups at the1193- or 4-position led to either a complete or substantial loss of120activity, as seen for compounds 5−8. Interestingly, by keeping121the 4-Cl group in place and removing the ethyl group on the122pyrazole to give 9, we observed a 10-fold improvement in the123inhibition of L4 development compared to TFP, furnishing a124single-digit nM IC50 value of 3 nM. A 3-fold improvement of125potency to inhibit L4 development was also observed in the126case of 10 compared to TFP, where the 4-Cl was replaced with1274-F. The results exhibited by 9 and 10 were encouraging, as128both the aims of increasing potency and partially reducing129hydrophobicity were attained. From our recent SAR study on a130broadly related 1-methyl-1H-pyrazole-5-carboxamide derived131from a different screening campaign against H. contortus, we132discovered that other five- or six-membered rings such as furan,133thiophene, substituted phenyl ring, or pyridinyl moiety were all

Figure 1. Structure, activity, and c Log P of the tolfenpyrad hit.

Scheme 1. Synthetic Pathway of the Tolfenpyrad Scaffolda

a(a) K2CO3, DMF; (b) LiAlH4, THF; and (c) HOAt, EDCI·HCl, ACN, or HATU, DIPEA, DMF, or T3P, DIPEA, THF.

Journal of Medicinal Chemistry Article

DOI: 10.1021/acs.jmedchem.8b01789J. Med. Chem. XXXX, XXX, XXX−XXX

B


134 disfavored on the RHS of the scaffold, and therefore are not135 explored in this study.19

136 The next aim was to explore SAR on the LHS of TFP, again137 with a focus on not only improving potency but also enhancing

t2 138 the physicochemical properties (Table 2). To increase139 hydrophilicity within the LHS region of TFP, the phenyl140 ring was replaced by the pyridine moiety, as demonstrated by141 11, 12, and 13. Gratifyingly, of these three pyridinyl142 compounds, while 11 completely lost activity, 12 and 13143 maintained potent inhibitory activity against L4 development144 with respective IC50 values of 0.03 and 0.019 μM, and a small145 loss of activity against xL3 motility.146 The roles of fluorine in medicinal chemistry are well147 established, in terms of enhancement of metabolic stability,148 potency, and permeability.20−22 Therefore, in addition to149 inclusion of a fluorine substituent in pyridinyl compound 13, a150 “fluorine walk” was undertaken for TFP itself, as testified by151 compounds 14−16. Here, it can be seen that the L3 activity152 was slightly weaker compared to TFP, but potent inhibitory153 activity on L4 development was maintained, in particular for154 15 and 16, for which the IC50 value of both was 0.04 μM.155 When the trifluoromethyl group was investigated, both the156 aromatic ring (17) and aliphatic chain (18) variants caused a157 complete loss of inhibitory activity. Heterocyclic N-oxides have158 been successfully used as therapeutic agents.23−25 For this159 reason, we synthesized and tested analogue 19, which harbors160 a pyridine N-oxide group. However, a substantial loss of161 activity against H. contortus was observed in relation to both162 xL3 motility and L4 development. Benzoxazole species, such as163 20, did not improve the original potency for either xL3 motility164 or L4 development, while carboxylic acid 21, a reported

165metabolite of TFP,10 was not tolerated. Activity was166maintained in both xL3 and L4 when the p-methyl group167was replaced with a p-chloro, as seen for 23, which exhibited a168potent L4 development IC50 value of 0.08 μM. Compound 22

Scheme 2. Synthetic Pathway Used for LHS Derivatizationb

b(a) HOAt, EDCI·HCl, ACN, or HATU, DIPEA, DMF, or T3P, DIPEA, THF; (b) CuI, Cs2CO3, N,N-dimethylglycine, 1,4-dioxane.

Table 1. SAR of the RHS Pyrazolea

entry RIC50 (μM) ± SD in xL3

motility assayIC50 (μM) ± SD in L4development assay

3 16.3 ± 6.66 0.13 ± 0.104 3-Et 10.53 ± 3.44 0.057 ± 0.0025 3-CN >1006 3-CF3 >1007 4-CN 50 ± 0.001 1.69 ± 0.678 4-CF3 >1009a 4-Cl 2.97 ± 2.56 0.003 ± 0.00410a 4-F 4.37 ± 2.55 0.01 ± 0.007TFP 2.9 ± 0.58 0.03 ± 0.005monepantel 0.16 ± 0.008 0.075 ± 0.04moxidectin 0.08 ± 0.04 3.45 ± 0.75

ac Log P: 9: 3.9; 10: 3.7.

Table 2. SAR of the LHS Region



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169 was synthesized as part of the SAR assessment and also for its

170 potential to serve as a probe for target identification due to the

171 azide-functional group. Click chemistry in activity-based

172 protein profiling for target identification has been extensively

173 reported in the literature.26−28 Although the azide-tagged

174 analogue 22 resulted in a dramatic motility reduction in xL3, it175 still displayed a binding affinity to inhibit L4 development,

176which suggests that azide-tagged TFP might find utility for177future target identification studies.178From the SAR investigation on the RHS of TFP, we179identified that the RHS of compounds 9 and 10 and the LHS180of 12 and 23 were optimal for the compounds tested. Having181successfully identified these groups, the focus then was on182incorporating them to develop a new set of analogues to probe183 t3the next generation of SAR, whose results are summarized in

Table 3. Next-Generation SAR on the LHS and RHS Regions of TFP

Table 4. Next-Generation SAR with 3-Methyl-4-chloropyrazole and 3-Methyl-4-fluoropyrazole RHS



D


t3 184 Table 3. For LHS with the pyridinyl moiety, compound 24,185 with the 4-chloropyrazole RHS, displayed similar activity in186 both xL3 and L4 compared to 12, whereas 28, with the 4-187 fluoropyrazole RHS, caused a moderate loss in activity.188 Excitingly, compounds 25 and 29 showed a substantial

189improvement in activity, reducing the IC50 value for xL3190motility and L4 development inhibition to submicromolar and191subnanomolar ranges, respectively. Furthermore, we also192extended the SAR scope for the LHS by testing the pyrimidine193(26 and 30) and 2-chloro pyridinyl (27 and 31) species.

Table 5. SAR on the Middle Region of TFP

Table 6. Next-Generation SAR on the LHS and Middle Region of TFP



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194 However, 26 and 30 caused a complete loss in activity, whereas195 no significant improvement in activity was observed for 27 and196 31. The loss in activity caused by 26 and 30 suggested a197 specific binding interaction exerted by the LHS region.198 From Table 3, the LHS groups that resulted in active199 compounds were selected for the development of a similar set200 of analogues, but with the 3-methyl-4-chloro and 3-methyl-4-201 fluoropyrazole RHS, as a complement to the 4-chloro and 4-

t4 202 fluoropyrazole RHS set (results summarized in Table 4). We203 included a 3-methyl group based on evidence already discussed204 for Table 1, which suggested that some hydrophobicity at this205 position might be favorable. Overall, similar activities against206 H. contortus xL3 motility and L4 development were observed207 within the two sets of analogues. In particular, compounds 32,208 35, and 37 maintained the activity originally observed for TFP.209 Compounds 33, 34, and 36 achieved an IC50 value in the210 subnanomolar range for L4 development inhibition.

211These encouraging results for the LHS and RHS of TFP212paved the way to explore SAR on the middle ring by testing the213fluoro substituent and the pyridinyl moiety. The results are214 t5summarized in Table 5. At the 2-position, the pyridinyl group215in compounds 38 produced a complete loss of activity, whereas216the fluoro substituent in 39 maintained the level of potency217observed for TFP against both xL3 motility and L4218development. Similar results were observed for the 3-position219(40 and 41).220From these results, it was decided to incorporate the fluoro-221substituted prototype, with the two previously identified222optimal LHS and the original RHS of TFP being kept223constant. This evaluation resulted in a set of analogues224 t6summarized in Table 6. When fluorine substitution was225explored at the 2-position, a 10-fold improvement in the226inhibition of L4 development was achieved for compound 42227compared to TFP, while 43 displayed the original L4228development activity. The originally observed levels of activity

Table 7. SAR on Various Linking Regions of the TFP Scaffold

entry R IC50 (μM) ± SD in xL3 motility assay

47 CH2 >10048 CO >10049 S >10050 N-Me >10051 >100

Table 8. Next-Generation SAR When Incorporating the Methylene Bridge into the TFP Scaffold



F


229 against xL3 by TFP were maintained for both 42 and 43.230 There was no significant improvement in the original activity231 against both xL3 and L4 when the same LHS groups were232 experimented upon with fluorine substitution at the 3-position233 of the middle ring, as seen for 44 and 45. Interestingly, a234 complete loss of activity against H. contortus was observed for235 compound 46 when an additional fluorine was implemented at236 the 5-position of the middle ring. These findings indicated a237 very tight SAR for this region.238 Alterations to the ether bridge and the benzylic carbon of239 TFP were also assessed. Different functional groups were used240 to replace the oxygen from the ether bridge, such as a

t7 241 methylene group (compound 47), a carbonyl group (48), at7 242 sulfur (49), or a methylated amine (50) (Table 7). However,

243 none of these replacement groups yielded active compounds. A244 similar result was achieved when the benzylic carbon was245 methylated to produce 51. Interestingly, when incorporating246 the methylene prototype into one of the four most active RHS

t8 247 pyrazoles to produce 52−55 (Table 8), activity was regained,248 but with no significant improvement with respect to TFP.249 From these results, the same RHS pyrazoles and the methylene250 bridge were then experimented upon with the active pyridinyl251 LHS, with the hope of achieving some improvement in activity.252 However, the original potency could not be maintained (56−253 59, Table 8).254 Having successfully identified compounds with significant255 improvement in activity compared to TFP, we selected nine256 compounds with high potency in inhibiting L4 development to257 test for their cytotoxicity on the MCF10A cell line, and the

t9 258 results are summarized in Table 9. We were delighted to

259observe high selectivity for the nine compounds tested,260particularly for compounds 27−29 and 31−34. Despite the261low cytotoxic IC50 value of 8.02 μM for one of our most potent262compounds 25, it was still a great level of selectivity compared263to the activity for L4 development of 0.7 nM. It was also264encouraging to see that our other most potent compounds 29265and 33 were not cytotoxic. These results reinforced the266potential of the TFP scaffold to be a novel scaffold with267anthelmintic activity.268To construct a biological activity profile for TFP and its269scaffold, we selected seven compounds, including TFP as a270control, to test for activity on three other parasitic nematodes271 t10at various concentrations (Table 10). The panel included272Ancylostoma ceylanicum (hookworm), Heligosomoides polygyrus273(rodent nematode), and T. muris (whipworm). It can be seen274that all seven compounds displayed 100% inhibition and >70%275inhibition of L3 of A. ceylanicum at 100 and 10 μM,276respectively. Similar results were seen for adult H. polygyrus,277where all compounds, except 33, showed complete inhibition278at 100 μM and >70% inhibition at only 1 μM. All compounds279exhibited >90% inhibition of first larval (L1) stage of T. muris280at 100 μM. There was an obvious improvement in the281inhibition of H. polygyrus L3 for the six newly developed282compounds compared to TFP.283In relation to the SAR for the previously reported pesticidal284activity by Okada et al.,12,29 while compounds 17, 48, and 49285were inactive against H. contortus in this study, they displayed286high potency against Nephotettix cincticeps (for 17) and both287Myzus persicae and Plutella xylostella (for 48 and 49). Likewise,288compound 4 with moderate activity against H. contortus was289not potent against Myzus persicae.14,29 These results indicated a290nonparallel SAR, in terms of inhibitory activity observed for H.291contortus in this study and the arthropod species studied by292Okada et al.14,29

293To assess the drug-likeliness of the TFP scaffold, we294 t11subjected TFP and nine other representative analogues (Table295 t1111) to different experiments, which determined key296physicochemical and metabolic parameters. The results297showed that all of our key compounds had reduced298hydrophobicity compared to TFP, including key high potency299compounds such as 34, which with a c Log P of 3.0 was 40300times less lipophilic than TFP (c Log P 4.6). This change, in301turn, led to an improvement in aqueous solubility at pH 6.5 for302all compounds, with some, such as 28, being around 20-fold303more soluble. At pH 2.0, solubility was also improved for most304compounds, except 23, 29, and 33. Concomitant with305decreased lipophilicity and improved solubility, all selected306compounds displayed a longer microsomal half-life than that

Table 9. Cytotoxicity Data for Selected Active Compoundson the MCF10A Cell Line

entryIC50 (μM) ± SD inxL3 motility assay

IC50 (μM) ± SD inL4 development assay

MCF10Acytotoxicity

IC50 (μM) ± SEM

TFP 2.57 ± 0.58 0.025 ± 0.005 37.90 ± 3.1110 4.37 ± 2.55 0.01 ± 0.007 13.13 ± 0.3823 2.43 ± 1.42 0.08 ± 0.006 8.8 ± 0.3425 0.38 ± 0.10 0.0007 ± 0 8.02 ± 0.4827 2.80 ± 2.26 0.02 ± 0.01 >5028 14.47 ± 7.45 0.19 ± 0.15 >5029 0.70 ± 0.24 0.0008 ± 0.0001 >5031 14.8 ± 9.6 0.17 ± 0.09 >5032 3.33 ± 1.89 0.01 ± 0.01 >5033 2.03 ± 1.82 0.0008 ± 0.0001 >5034 1.8 ± 0.49 0.008 ± 0.009 >50

Table 10. Biological Activity Profile of Selected Compounds against a Panel of Parasitic Nematodes

H. polygyrus adult(% inhibition) H. polygyrus L3 (% inhibition) A. ceylanicum L3 (% inhibition) T. muris L1 (% inhibition)

entry 10 μM 1 μM 100 μM 100 μM 10 μM 100 μM

TFP 100 73.4 32.4 100 94.85 10010 100 85.9 91.8 100 100 91.9723 100 100 89.2 100 98.8 10027 100 100 100 100 73.15 10029 100 90.6 93.7 100 98.8 93.4533 96.5 96.9 66.9 100 81.6 10034 100 100 99.2 100 85.4 100



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307 observed for TFP, ranging from 20 min for 10 to 84 min for308 27.

309 ■ CONCLUSIONS310 We have discovered that TFP potently inhibits the develop-311 ment of L4 stage larvae of the ovine parasitic nematode, H.312 contortus. Herein, we report a systematic SAR interrogation313 that has led to the identification of novel modifications that not314 only improve druglike physicochemical properties, such as315 lipophilicity, aqueous solubility, and microsomal degradation316 half-life, but are also exquisitely potent. For example, 25, 29,317 and 33 achieved a remarkable improvement in inhibitory318 activity against both xL3 motility and L4 development,319 reducing the original and already impressive IC50 values of320 TFP down to the submicromolar or subnanomolar ranges, at321 the same time, maintaining selectivity toward the parasite.322 TFP, as a pesticide, was reported to be a complex I inhibitor323 that disrupts the respiratory electron transport chain in324 mitochondria.11,30 However, the specific biological target(s)325 of this series of compounds in nematodes is/are unknown.326 With a moderately potent inhibitory affinity observed against327 L4 development, compound 22 might represent a potential328 tool for target identification. Having established a compre-329 hensive antiparasitic SAR herein, we are now embarking on330 pathway identification and further downstream efficacy assess-331 ment and hope to report on these efforts in due course.

332 ■ EXPERIMENTAL SECTION333 The nematode assays and cytotoxicity assay are as described by Le et334 al.19

335 ■ PHYSICOCHEMICAL EXPERIMENTAL336 Calculated Physicochemical Parameters Using ChemAxon337 JChem Software. A range of physicochemical properties evaluating338 drug-likeliness and likely oral absorption characteristics were339 calculated using the ChemAxon chemistry cartridge via JChem for340 Excel software (version 16.4.11). A brief description of each341 parameter is provided: c Log P: partition coefficients, reflecting the342 lipophilic character of the neutral structure.343 Kinetic Solubility Estimation Using Nephelometry (SolpH).344 Compound in DMSO was spiked into either pH 6.5 phosphate buffer345 or 0.01 M HCl (approximately pH 2.0) with the final DMSO346 concentration being 1%. After 30 min had elapsed, the samples were347 then analyzed via nephelometry to determine a solubility range (see348 Bevan et al.31).31

349 In Vitro Metabolic Stability. The procedure is as described by350 Le et al.19

351■ CHEMISTRY EXPERIMENTAL352All solvents and reagents were used directly from commercial353suppliers unless otherwise stated. All of the final compounds had354purities greater than 95% based on analytical HPLC, 1H NMR, and355LC-MS methods. General chemistry experimental conditions were as356reported by Le et al.19

357General Procedure A1: Nucleophilic Aromatic Substitution358Reactions. To a stirred solution of arylfluoride (1.0 equiv) and359nucleophile (1.1 equiv) in DMF (20 mL) was added Cs2CO3 (2.0360equiv), and the mixture was stirred at 100 °C for 2 h. Upon361completion, the reaction mixture was diluted with EtOAc (50 mL)362and organic layer was washed with water and brine, dried over363anhydrous MgSO4, and concentrated in vacuo. Crude product was364purified by column chromatography (5−20% EtOAc/petroleum365benzine) to yield the desired product.366General Procedure A2: Nucleophilic Aromatic Substitution367Reactions. To a stirred solution of arylfluoride (1.0 equiv) and368nucleophile (1.0 equiv) in DMF (20 mL), K2CO3 (2.0 equiv) was369added. The reaction mixture was then stirred at 100 °C overnight.370Upon completion, the reaction mixture was extracted with EtOAc,371washed with water and brine, dried over anhydrous Na2SO4, and372concentrated in vacuo. Crude product was purified by column373chromatography (5−20% EtOAc/petroleum benzine) to yield the374desired product.375General Procedure B1: Nitrile Reduction. LiAlH4 (3.0 equiv)376was slowly added to a solution of substituted benzonitrile (1.0 equiv)377in anhydrous THF (10 mL). The reaction was stirred at room378temperature for 2 h and then cooled on ice before a solution of 1 M379NaOH was added. The slurry mixture was then filtered through a pad380of celite and filtrate was extracted with EtOAc (3 × 30 mL).381Combined organic layers were dried (MgSO4) and concentrated in382vacuo. HCl (4 M) in 1,4-dioxane (15 mL) was added to the residue,383and then the reaction mixture was stirred at room temperature384overnight. Precipitate of the resulting HCl salt was then filtered,385washed with diethyl ether, and dried in a vacuum oven to yield the386desired product as an HCl salt.387General Procedure B2: Nitrile Reduction. A stirred solution of388substituted benzonitrile (2.39 mmol) in THF (10 mL) was cooled to3890 °C and LiAlH4 solution (2.4 mL, 2 M in THF) was added dropwise390under N2. The reaction mixture was stirred at room temperature for 3391h and then quenched with saturated Na2SO4 solution dropwise. The392slurry mixture was filtered through a pad of celite and washed393thoroughly with EtOAc. The filtrate was washed with water and brine,394dried over anhydrous Na2SO4, and concentrated in vacuo to yield the395desired benzylamine, which was taken to the next step without any396purification.397General Procedure B3: Nitrile Reduction. To a stirred solution398of substituted benzonitrile (1.06 mmol) in MeOH (10 mL) was399added Raney Ni (1.16 mmol), followed by NH3 (1 mL, 7.0 M in400MeOH). The reaction mixture was stirred at room temperature for 3401h. Upon completion, the reaction mixture was filtered through a pad

Table 11. Key Physicochemical Parameters and in Vitro Metabolic Stability of Selected Compounds

c Log Pa solb (μg/mL)

ID pH 2.0 pH 6.5 T1/2 (min) CLint, in vitroc (μL/min/mg protein) microsome-predicted EH

d

TFP 4.6 3.1−6.3 <1.6 14 121 N/A10 3.3 25−50 6.3−2.5 20 87 0.6523 4.1 1.6−3.1 1.6−3.1 36 48 0.5127 2.8 6.3−12.5 6.3−12.5 84 21 0.3128 1.7 >100 25−50 25 69 0.6029 3.4 3.1−6.3 1.6−3.1 57 30 0.3931 2.4 9−18 9−18 80 22 0.3232 2.3 >100 12.5−25 31 55 0.5433 4.0 1.6−3.1 3.1−6.3 80 22 0.3234 3.0 4.9−9.8 2.4−4.9 55 31 0.40

aCalculated using ChemAxon JChem software. bKinetic solubility determined by nephelometry (SolpH).cIn vitro intrinsic clearance determined in

mouse liver microsomes. dPredicted hepatic extraction ratio calculated from in vitro data.



H


402 of celite and the filtrate was concentrated in vacuo to yield the desired403 benzylamine, which was taken directly to the next step without further404 purification.405 General Procedure B4: Nitrile Reduction. Substituted406 benzonitrile (1.0 equiv), di-tert-butyl dicarbonate (1.5 equiv), and407 NiCl2·6H2O (0.2 equiv) were dissolved in anhydrous MeOH in an408 oven-dried flask under N2. The mixture was cooled to 0 °C before409 NaBH4 (7.0 equiv) was added in small portions over 20 min. The410 reaction mixture was stirred at room temperature for 2 h.411 Diethylenetriamine (1.0 equiv) was then added, and the reaction412 mixture was stirred for another 15 min before MeOH was removed in413 vacuo. Saturated solution of NaHCO3 was added to the residue and414 extracted with EtOAc (3 × 20 mL). Combined organic layers were415 washed with water and brine, dried (MgSO4), and solvent was416 removed in vacuo to afford the desired Boc-protected benzylamine,417 which was then stirred in 4 M solution of HCl in 1,4-dioxane at 60 °C418 for 2 h to yield the corresponding HCl salt upon filtration of419 precipitate.420 General Procedure B5: Nitrile Reduction. Di-tert-butyl421 dicarbonate (2.0 equiv) and NiCl2·6H2O (0.2 equiv) were added to422 a stirred solution of substituted benzonitrile (1.0 equiv) in MeOH (7423 mL) at 0 °C. NaBH4 (7.0 equiv) was then added to the reaction424 mixture in small portions over 30 min. The reaction mixture was425 stirred at room temperature for 1 h before it was filtered through a426 pad of celite. The filtrate was diluted with EtOAc, washed with a427 saturated NaHCO3 solution and brine, dried over Na2SO4, and428 concentrated in vacuo to yield crude product without further429 purification.430 General Procedure C1: Amide Coupling. Amine (either as free431 base or HCl salt, 1.0 equiv), HOAt (2.0 equiv), Et3N (2.0 equiv),432 EDCI·HCl (2.0 equiv), and carboxylic acid (2.0 equiv) were dissolved433 in 3 mL of DMF. The reaction mixture was heated at 80 °C until434 completion before EtOAc was added. The organic layer was washed435 with water, dried (MgSO4), and solvent was removed in vacuo to give436 crude product, which was purified by column chromatography (5−437 10% EtOAc/petroleum benzine) to yield the desired product.438 General Procedure C2: Amide Coupling. T3P (2.0 equiv, 50%439 in EtOAc) and DIPEA (3.0 equiv) were added to a stirred solution of440 carboxylic acid (1.0 equiv) and amine (1.0 equiv) in THF (5 mL).441 The reaction mixture was stirred at room temperature for 6 h before442 EtOAc was added. The organic layer was washed with saturated443 NaHCO3 solution and brine, dried over Na2SO4, and concentrated in444 vacuo to give crude product, which was purified by prep-HPLC to445 afford the desired product.446 General Procedure C3: Amide Coupling. HATU (0.97 mmol)447 and DIPEA (1.28 mmol) were added to a stirred solution of448 carboxylic acid (0.54 mmol) in DMF (5.0 mL). The solution was449 stirred for 5 min before amine (0.71 mmol) was added, and the450 reaction was stirred further at room temperature for 16 h. Upon451 completion, the reaction was diluted with EtOAc, washed with water452 and brine, and then dried over anhydrous Na2SO4. Solvent was then453 removed in vacuo to give crude product, which was purified by454 column chromatography (5−20% EtOAc/petroleum benzine) to455 yield the desired product.456 General Procedure C4: Amide Coupling. EDCI·HCl (1.2457 equiv) and HOAt (1.2 equiv) were added to a solution of carboxylic458 acid (1 equiv) in ACN (0.8 M) at room temperature. The reaction459 mixture was heated to 50 °C before amine (1.2 equiv) was added after460 10 min. The reaction was stirred at this temperature overnight before461 it was cooled to room temperature and concentrated in vacuo. The462 residue was extracted with EtOAc (2 × 10 mL) and washed with463 water. Combined organic layers were dried over MgSO4 and then464 loaded directly onto silica. The crude product was purified by silica465 gel chromatography (Isolera Biotage, 0−50% EtOAc/petroleum466 benzine). Product-containing fractions were combined and concen-467 trated in vacuo to give the desired product.468 General Procedure C5: Amide Coupling. To a stirred solution469 of carboxylic acid (1.0 equiv) in pyridine (5 mL), T3P (50% in470 EtOAc, 7.0 equiv) was added and the reaction mixture was stirred for471 5 min before amine (free base or HCl salt, 1.0 equiv) was added. The

472reaction mixture was stirred at room temperature for 16 h. Upon473completion, the reaction mixture was diluted with EtOAc. The474organic layer was washed with saturated NaHCO3 solution and brine,475dried over anhydrous Na2SO4, and solvent was concentrated in vacuo.476Crude product was purified by column chromatography (5−20%477EtOAc/petroleum benzine) to yield the desired product.478General Procedure C6: Amide Coupling. Carboxylic acid479(14.58 mmol), amine HCl (16.04 mmol), EDCI·HCl (16.04 mmol),480and Et3N (32.08 mmol) were dissolved in dichloromethane (DCM).481The reaction mixture was stirred at room temperature overnight.482Upon completion, DCM was removed in vacuo and the residue was483extracted with EtOAc, washed with water and brine, dried (MgSO4),484and concentrated in vacuo to afford crude product, which was purified485by column chromatography to yield the desired product.486General Procedure D1: Ester Hydrolysis. LiOH·H2O (2.0487equiv) was added to a stirred solution of ester (1.0 equiv) in a 3:1488mixture of THF/H2O. The reaction mixture was stirred at room489temperature for 3 h before THF was removed in vacuo. Aqueous layer490was then neutralized by 1 N HCl and extracted with EtOAc, washed491with brine, dried over anhydrous Na2SO4, and concentrated in vacuo492to afford the desired carboxylic acid without further purification.493General Procedure D2: Ester Hydrolysis. NaOH (23.75 mmol)494was added to a solution of ester (1.04 mmol) in EtOH (10 ml). The495mixture was stirred at room temperature overnight before solvent was496removed in vacuo. The residue was redissolved in water and washed497with EtOAc (3 × 10 mL). The aqueous layer was acidified with 1 M498HCl to pH ∼ 3 and extracted with EtOAc (3 × 10 mL). Combined499organic layers were dried (MgSO4), and solvent was removed in500vacuo to give the desired carboxylic acid without further purification.501General Procedure D3: Ester Hydrolysis. LiOH (5.0 equiv)502was added to a solution of ester (1.0 equiv) in THF (10 mL). The503reaction was stirred at room temperature overnight before THF was504removed in vacuo. Water (20 mL) was then added to the residue,505followed by 1 M HCl to pH ∼ 1. The aqueous was extracted with506EtOAc (2 × 10 mL). Combined organic layers was washed with brine,507dried (MgSO4), and concentrated in vacuo to give the desired508carboxylic acid without further purification.509General Procedure E1: Ullmann-Type Coupling. In a510microwave tube charged with a magnetic stirrer bar, arylhalide (1.0511equiv), substituted phenol (1.5 equiv), CuI (0.1 equiv), Cs2CO3 (2.0512equiv), and N,N-dimethylglycine (0.4 equiv) were dissolved in 1,4-513dioxane. The tube was sealed with a cap and placed in a microwave514reactor heated at 110 °C for 2 h. Upon completion, the reaction515mixture was diluted with EtOAc, washed with H2O and brine, dried516(MgSO4), and solvent was removed in vacuo to afford crude product.517Crude was purified by column chromatography (5−20% EtOAc/518petroleum benzine) to yield the desired product.519General Procedure E2: Ullmann-Type Coupling. Substituted520thiophenol (4.03 mmol), K2CO3 (8.05 mmol), and arylhalide (6.04521mmol) were dissolved in toluene (5 mL) in a sealed tube. The522mixture was degassed for 0.5 h before 1,10-phenanthroline (0.4523mmol) and CuI (0.4 mmol) were added. The reaction mixture was524stirred at 130 °C for 2 h. Upon completion, the reaction mixture was525filtered through a pad of celite. The filtrate was extracted with EtOAc526(3 × 20 mL), and the combined organic layers were washed with527water and brine, dried (MgSO4), and concentrated in vacuo. Crude528product was purified by column chromatography (5−20% EtOAc/529petroleum benzine) to yield the desired product.530General Procedure F: Primary Amine Synthesis from531Aldehyde. Substituted benzaldehyde (1.0 equiv) was dissolved in532EtOH (10 mL), NH2OH·HCl (1.2 equiv) was then added, and the533reaction mixture was stirred at room temperature for 1 h before 3 mL534of concentrated HCl (37%) was added, followed by Zn dust (2.5535equiv). The reaction was stirred for another 15 min before basified536with excess amount of aqueous NH3 and 6 M NaOH solution. The537resulting slurry mixture was filtered through a pad of celite and diluted538with EtOAc. The organic was washed with water, dried (MgSO4), and539concentrated in vacuo to afford the desired benzylamine as a free base,540which was carried through to the next step without any purification.



I


541 General Procedure G1: Chan−Lam coupling. To a stirred542 solution of substituted phenol (3.01 mmol) in DCM (10 mL), aryl543 boronic acid (9.03 mmol) was added. Et3N (6.02 mmol) and544 Cu(OAc)2 (3.01 mmol) were then added and the reaction mixture545 was degassed for 5 min. The reaction mixture was stirred under O2546 balloon for 18 h. Upon completion, the reaction mixture was filtered547 and residue was diluted with DCM. Organic layer was washed with548 cold water and brine, dried over anhydrous Na2SO4, and concentrated549 in vacuo to give the desired product, which was used in the next step550 without any purification.551 General Procedure G2: Chan−Lam Coupling. To a stirred552 solution of substituted phenol (1.0 equiv), aryl boronic acid (2.0553 equiv), and Et3N (5.0 equiv) in DCM (30 mL) was added Cu(OAc)2554 (1.0 equiv), followed by 4 Å molecular sieves (0.5 g). The reaction555 mixture was then stirred under O2 for 16 h. Upon completion, the556 reaction mixture was filtered through a pad of celite and the filtrate557 was washed with 10% aqueous NaHSO4 solution and 1 N NaOH558 solution. The organic layer was extracted and dried over anhydrous559 Na2SO4 and then concentrated in vacuo to give crude product, which560 was purified by column chromatography (5−20% EtOAc/petroleum561 benzine) to yield the desired product.562 General Procedure H1: Weinreb Ketone Synthesis. Weinreb563 amide (0.3 mmol) was dissolved in anhydrous THF (5 mL) in an564 oven-dried round-bottom flask under N2. The mixture was cooled in565 an ice bath to 0 °C before Grignard reagent (0.7 mmol) was added.566 The reaction mixture was stirred at room temperature for 1 h. Upon567 completion, THF was removed in vacuo and saturated NH4Cl568 solution was added to the residue, which was then extracted with569 EtOAc (3 × 10 mL). Combined organic layers were washed with570 water and brine, dried (MgSO4), and concentrated in vacuo to give571 crude product, which was purified by column chromatography (5−572 20% EtOAc/petroleum benzine) to yield the desired product.573 General Procedure H2: Weinreb Ketone Synthesis. Arylha-574 lide (24.4 mmol) was dissolved in anhydrous THF (15 mL) in an575 oven-dried round-bottom flask under N2. The mixture was cooled to576 −78 °C before nBuLi (2.5 M in hexane, 24.4 mmol) was added577 dropwise, and the mixture was left stirred for 30 min. A solution of578 Weinreb amide (12.2 mmol) in THF (10 mL) was then added. The579 reaction mixture was stirred at −78 °C for 45 min. Upon completion,580 water was added to quench the reaction, followed by extraction with581 EtOAc (3 × 20 mL). Combined organic layers were washed with582 brine, dried (MgSO4), and concentrated in vacuo to give crude583 product, which was purified by column chromatography (5−20%584 EtOAc/petroleum benzine) to yield the desired product.585 4-(p-Tolyloxy)benzonitrile (1). Title compound was prepared586 according to General procedure A1, starting from p-cresol and 4-587 fluorobenzonitrile to give a colorless oil (1.9 g, 37%). 1H NMR (400588 MHz, CDCl3): δ = 7.56 (d, J = 8.8 Hz, 2H), 7.19 (d, J = 8.3 Hz, 2H),589 6.98−6.93 (m, 4H), 2.36 (s, 3H) ppm; LC-MS: m/z = 210.3 [M +590 H]+.591 (4-(p-Tolyloxy)phenyl)methanamine (2). Title compound was592 prepared according to General procedure B2, starting from 1 to give a593 colorless oil (78%). 1H NMR (400 MHz, CDCl3): δ = 7.23−7.26 (m,594 2H), 7.1 (d, J = 8.1 Hz, 2H), 6.94 (d, J = 8.4 Hz, 2H), 6.89 (d, J = 8.3595 Hz, 2H), 3.83 (br, 2H), 2.32 (s, 3H) ppm. LC-MS: m/z = 214.3 [M +596 H]+.597 1-Methyl-N-(4-(p-tolyloxy)benzyl)-1H-pyrazole-5-carboxa-598 mide (3). Title compound was prepared according to General599 procedure C1, starting from 2 and 1-methyl-1H-pyrazole-5-carboxylic600 acid to give a brown oil (74%). 1H NMR (400 MHz, CDCl3) δ = 7.31601 (s, J = 1.8 Hz, 1H), 7.16 (d, J = 8.4 Hz, 2H), 7.05 (d, J = 8.5 Hz, 2H),602 6.88−6.77 (m, 4H), 6.54 (s, br, 1H), 6.43 (d, J = 2.1 Hz, 1H), 4.43603 (d, J = 5.7 Hz, 2H), 4.08 (s, 3H), 2.24 (s, 3H) ppm; 13C NMR (101604 MHz, CDCl3) δ = 159.9, 157.6, 154.5, 137.6, 135.1, 133.2, 132.1,605 130.3, 129.3, 119.2, 118.5, 106.4, 43.0, 39.3, 20.7 ppm; LC-MS: m/z606 = 321.9 [M + H]+.607 3-Ethyl-1-methyl-N-(4-(p-tolyloxy)benzyl)-1H-pyrazole-5-608 carboxamide (4). Title compound was prepared according to609 General procedure C1, starting from 2 and 3-ethyl-1-methyl-1H-610 pyrazole-5-carboxylic acid to give a colorless oil (21%). 1H NMR

611(400 MHz, CDCl3) δ = 7.20−7.00 (m, 4H), 6.93−6.77 (m, 4H), 6.32612(s, br, 1H), 6.21 (d, J = 7.0 Hz, 1H), 4.43 (d, J = 6.0 Hz, 2H), 4.03 (d,613J = 7.1 Hz, 4H), 2.54−2.48 (m, 2H), 2.24 (d, J = 6.8 Hz, 3H), 1.13 (t,614J = 7.6 Hz, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ = 160.0, 157.5,615154.5, 153.00, 135.6, 133.2, 132.1, 130.4, 129.3, 119.2, 118.6, 104.2,61643.0, 38.9, 21.2, 20.8, 13.9 ppm; LC-MS: m/z = 349.9.6173-Cyano-1-methyl-N-(4-(p-tolyloxy)benzyl)-1H-pyrazole-5-618carboxamide (5). Title compound was prepared according to619General procedure C2, starting from 2 and 65 to give an off-white620solid (26%). 1H NMR (400 MHz, DMSO-d6): δ = 9.25 (t, J = 5.8 Hz,6211H), 7.50 (s, 1H), 7.31 (d, J = 8.5 Hz, 2H), 7.18 (d, J = 8.2 Hz, 2H),6226.93 (d, J = 8.5 Hz, 2H), 6.88 (d, J = 8.4 Hz, 2H), 4.42 (d, J = 5.8 Hz,6232H), 4.15 (s, 3H), 2.28 (s, 3H) ppm; LC-MS: m/z = 347.1 [M + H]+.6241-Methyl-N-(4-(p-tolyloxy)benzyl)-3-(trifluoromethyl)-1H-625pyrazole-5-carboxamide (6). Title compound was prepared626according to General procedure C3, starting from 2 and 1-methyl-6273-(trifluoromethyl)-1H-pyrazole-5-carboxylic acid to give a yellow628solid (60%). 1H NMR (400 MHz, DMSO-d6): δ = 9.20 (t, J = 5.6 Hz,6291H), 7.35 (s, 1H), 7.31 (d, J = 8.5 Hz, 2H), 7.19 (d, J = 8.2 Hz, 2H),6306.93 (d, J = 8.4 Hz, 2H), 6.89 (d, J = 8.3 Hz, 2H), 4.42 (d, J = 5.6 Hz,6312H), 4.14 (s, 3H), 2.27 (s, 3H); ppm; LC-MS: m/z = 390.1 [M +632H]+.6334-Cyano-1-methyl-N-(4-(p-tolyloxy)benzyl)-1H-pyrazole-5-634carboxamide (7). To a stirred solution of 4-iodo-1-methyl-1H-635pyrazole-5-carboxylic acid methyl ester (700 mg, 2.63 mmol) in DMF636(10 mL), CuCN (472 mg, 5.26 mmol) was added. The reaction was637stirred at 140 °C for 3 h. Upon completion of the reaction, the638reaction mixture was cooled to room temperature and extracted with639EtOAc; washed with saturated NH4Cl, water, and brine; dried over640anhydrous Na2SO4; and concentrated in vacuo. Crude product was641purified by column chrom atography (30% EtOAc in hexane) to642afford methy-4-cyano-1-methyl-1H-pyrazole-5-carboxylate as a white643solid, which was directly subjected to General procedure D1 to give644the corresponding carboxylic acid that was subsequently coupled to64559 according to General procedure C2 to give the title compound as646an off-white solid (68%). 1H NMR (400 MHz, DMSO-d6): δ = 9.48647(br, 1H), 8.12 (s, 1H), 7.36 (d, J = 8.1 Hz, 2H), 7.19 (d, J = 7.9 Hz,6482H), 6.95−6.89 (m, 4H), 4.47 (d, J = 5.4 Hz, 2H), 3.95 (s, 3H), 2.29649(s, 3H) ppm; LC-MS: m/z = 347.2 [M + H]+.6501-Methyl-N-(4-(p-tolyloxy)benzyl)-4-(trifluoromethyl)-1H-651pyrazole-5-carboxamide (8). Title compound was prepared652according to General procedure C3, starting from 2 and 1-methyl-6534-(trifluoromethyl)-1H-pyrazole-5-carboxylic acid to give a white654solid (53%). 1H NMR (400 MHz, CDCl3): δ = 7.65 (s, 1H), 7.26 (d,655J = 8.4 Hz, 2H), 7.13 (d, J = 8.3 Hz, 2H), 6.95 (d, J = 8.5 Hz, 2H),6566.90 (d, J = 8.4 Hz, 2H), 6.41 (d, J = 5.6 Hz, 1H), 4.57 (d, J = 5.6 Hz,6572H), 4.09 (s, 3H), 2.32 (s, 3H) ppm; LC-MS: m/z = 390.1 [M + H]+.6584-Chloro-1-methyl-N-(4-(p-tolyloxy)benzyl)-1H-pyrazole-5-659carboxamide (9). Title compound was prepared according to660General procedure C1, starting from (4-(4-chlorophenoxy)phenyl)-661methanamine HCl and 4-chloro-1-methyl-1H-pyrazole-5-carboxylic662acid to give a white solid (19%). 1H NMR (400 MHz, CDCl3) δ =6637.43 (s, 1H), 7.31−7.27 (m, 2H), 7.14 (d, J = 8.2 Hz, 2H), 7.01−6.88664(m, 5H), 4.60 (d, J = 5.7 Hz, 2H), 4.19 (s, 3H), 2.33 (s, 3H) ppm;665

13C NMR (101 MHz, CDCl3) δ = 158.3, 157.6, 154.5, 136.7, 133.3,666131.7, 131.0, 130.4, 129.2, 119.3, 118.6, 109.6, 43.1, 41.2, 20.8 ppm;667LC-MS: m/z = 355.8 [M + H]+.6684-Fluoro-1-methyl-N-(4-(p-tolyloxy)benzyl)-1H-pyrazole-5-669carboxamide (10). Title compound was prepared according to670General procedure C4, starting from 2 and 67 to give a white solid671(62% yield). 1H NMR (400 MHz, CDCl3) δ = 7.36 (d, J = 4.5 Hz,6721H), 7.34−7.27 (m, 1H), 7.16 (d, J = 8.1 Hz, 2H), 7.03−6.90 (m,6734H), 6.51 (s, 1H), 4.60 (d, J = 5.7 Hz, 2H), 4.19 (d, J = 0.9 Hz, 3H),6742.36 (s, 3H) ppm; LC-MS m/z = 339.9 [M + H]+.6754-Chloro-3-ethyl-1-methyl-N-(4-((5-methylpyridin-2-yl)-676oxy)benzyl)-1H-pyrazole-5-carboxamide (11). General proce-677dure C1 was followed, starting from 4-chloro-3-ethyl-1-methyl-1H-678pyrazole-5-carboxylic acid and (4-bromophenyl)methanamine HCl to679give N-(4-bromobenzyl)-4-chloro-3-ethyl-1-methyl-1H-pyrazole-5-680carboxamide, which was subsequently coupled to 5-methylpyridin-2-



J


681 ol according to General procedure E1 to give the title compound as a682 white solid (20% yield). 1H NMR (400 MHz, CDCl3) δ = 7.46 (d, J =683 8.2 Hz, 2H), 7.36 (d, J = 8.3 Hz, 2H), 7.27 (dd, J = 9.4, 2.4 Hz, 1H),684 7.22 (s, br, 1H), 7.10 (s, 1H), 6.60 (d, J = 9.3 Hz, 1H), 4.67 (d, J =685 5.8 Hz, 2H), 4.13 (s, 3H), 2.64 (q, J = 7.6 Hz, 2H), 2.10 (s, 3H), 1.24686 (t, J = 6.9 Hz, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ = 161.8,687 158.7, 149.6, 142.8, 140.4, 138.0, 135.2, 131.0, 128.5, 127.0, 121.4,688 115.2, 107.8, 42.9, 40.6, 19.2, 17.0, 12.8 ppm; LC-MS m/z = 384.8689 [M + H]+.690 4-Chloro-3-ethyl-1-methyl-N-(4-((6-methylpyridin-3-yl)-691 oxy)benzyl)-1H-pyrazole-5-carboxamide (12). General proce-692 dure C1 was followed, starting from 4-chloro-3-ethyl-1-methyl-1H-693 pyrazole-5-carboxylic acid and (4-bromophenyl)methanamine HCl to694 give N-(4-bromobenzyl)-4-chloro-3-ethyl-1-methyl-1H-pyrazole-5-695 carboxamide, which was subsequently coupled to 6-methylpyridin-2-696 ol according to General procedure E1 to give the title compound as a697 white solid (12% yield). 1H NMR (400 MHz, CDCl3) δ = 8.27 (d, J =698 2.7 Hz, 1H), 7.32 (d, J = 8.7 Hz, 2H), 7.22 (dd, J = 8.5, 2.8 Hz, 1H),699 7.12 (d, J = 8.5 Hz, 1H), 7.05 (s, br, 1H), 6.98−6.94 (m, 2H), 4.60700 (d, J = 5.8 Hz, 2H), 4.13 (s, 3H), 2.62 (q, J = 7.6 Hz, 2H), 2.53 (s,701 3H), 1.22 (t, J = 7.6 Hz, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ =702 158.6, 156.7, 153.5, 151.4, 149.7, 140.8, 132.9, 131.0, 129.4, 126.9,703 123.9, 118.7, 107.7, 42.9, 40.7, 23.6, 19.3, 12.9 ppm; LC-MS m/z =704 384.8 [M + H]+.705 4-Chloro-3-ethyl-N-(4-((2-fluoro-6-methylpyridin-3-yl)oxy)-706 benzyl)-1-methyl-1H-pyrazole-5-carboxamide (13). General707 procedure A1 was followed, starting from 2-fluoro-6-methylpyridin-708 3-ol and 4-fluorobenzaldehyde to give 4-((2-fluoro-6-methylpyridin-3-709 yl)oxy)benzaldehyde, which was converted to the corresponding710 benzylamine according to General procedure F, and then711 subsequently coupled to 4-chloro-3-ethyl-1-methyl-1H-pyrazole-5-712 carboxylic acid according to General procedure C1 to give the title713 compound as a brown solid (46% yield). 1H NMR (400 MHz,714 CDCl3) δ = 7.36−7.28 (m, 3H), 7.03 (s, br, 1H), 6.98 (d, J = 7.9 Hz,715 1H), 6.95−6.90 (m, 2H), 4.59 (d, J = 5.8 Hz, 2H), 4.11 (s, 3H), 2.60716 (q, J = 7.6 Hz, 2H), 2.46 (s, 3H), 1.21 (t, J = 7.6 Hz, 3H) ppm; 13C717 NMR (101 MHz, CDCl3) δ = 158.6, 156.4, 155.7, 153.3, 151.6 (d, J =718 12.5 Hz), 149.6, 136.3 (d, J = 26.5 Hz), 132.9, 131.4 (d, J = 3.7 Hz),719 131.0, 129.3, 121.4 (d, J = 4.5 Hz), 117.6, 107.7, 42.9, 40.6, 23.2, 19.3,720 12.8 ppm; LC-MS m/z = 402.8 [M + H]+.721 4-Chloro-3-ethyl-N-(4-(3-fluoro-4-methylphenoxy)benzyl)-722 1-methyl-1H-pyrazole-5-carboxamide (14). Title compound was723 prepared according to General procedure C2, starting from 68 and 4-724 chloro-3-ethyl-1-methyl-1H-pyrazole-5-carboxylic acid to give an off-725 white solid (25%). 1H NMR (400 MHz, DMSO-d6): δ 8.92 (t, J = 5.6726 Hz, 1H), 7.37 (d, J = 8.3 Hz, 2H), 7.27 (t, J = 8.5 Hz, 1H), 7.01 (d, J727 = 8.3 Hz, 2H), 6.82 (d, J = 10.9 Hz, 1H), 6.73 (d, J = 6.3 Hz, 1H),728 4.46 (d, J = 5.6 Hz, 2H), 3.84 (s, 3H), 2.57-2.53 (m, 2H), 2.19 (s,729 3H), 1.16 (t, J = 7.4 Hz, 3H) ppm; LC-MS: m/z = 402.1 [M + H]+.730 4-Chloro-3-ethyl-N-(4-(2-fluoro-4-methylphenoxy)benzyl)-731 1-methyl-1H-pyrazole-5-carboxamide (15). Title compound was732 prepared according to General procedure C2, starting from 69 and 4-733 chloro-3-ethyl-1-methyl-1H-pyrazole-5-carboxylic acid to give a color-734 less gum (21%). 1H NMR (400 MHz, DMSO-d6): δ 8.90 (t, J = 5.8735 Hz, 1H), 7.32 (d, J = 8.5 Hz, 2H), 7.21 (d, J = 11.6 Hz, 1H), 7.09−736 7.02 (m, 2H), 6.91 (d, J = 8.4 Hz, 2H), 4.43 (d, J = 5.9 Hz, 2H), 3.83737 (s, 3H), 2.57−2.53 (m, 2H), 2.32 (s, 3H), 1.16 (t, J = 7.5 Hz, 3H)738 ppm; LC-MS: m/z = 402.1 [M + H]+.739 4-Chloro-3-ethyl-N-(4-(4-fluorophenoxy)benzyl)-1-methyl-740 1H-pyrazole-5-carboxamide (16). General procedure B2 was741 followed, starting from 70 to give (4-(4-fluorophenoxy)phenyl)-742 methanamine, which was subsequently coupled to 4-chloro-3-ethyl-1-743 methyl-1H-pyrazole-5-carboxylic acid according to General procedure744 C2 to give the title compound as a light yellow gum (37%). 1H NMR745 (400 MHz, DMSO-d6): δ 8.91 (s. br, 1H), 7.35 (d, J = 8.3 Hz, 2H),746 7.22 (t, J = 8.8 Hz, 2H), 7.06−7.03 (m, 2H), 6.97 (d, J = 8.3 Hz, 2H),747 4.45 (d, J = 5.8 Hz, 2H), 3.83 (s, 3H), 2.57−2.53 (m, 2H), 1.16 (t, J =748 7.4 Hz, 3H) ppm; LC-MS: m/z = 388.1 [M + H]+.749 4-Chloro-3-ethyl-1-methyl-N-(4-(4-(trifluoromethyl)-750 phenoxy)benzyl)-1H-pyrazole-5-carboxamide (17). General

751procedure A2 was followed, starting from 4-(trifluoromethyl)phenol752and 4-fluorobenzonitrile to give 4-(4-(trifluoromethyl)phenoxy)-753benzonitrile, which was then reduced according to General procedure754B2 to give the corresponding benzylamine that was subsequently755coupled to 4-chloro-3-ethyl-1-methyl-1H-pyrazole-5-carboxylic acid756according to General procedure C3 to give the title compound as a757white solid (19%). 1H NMR (400 MHz, DMSO-d6): δ = 8.95 (t, J =7585.7 Hz, 1H), 7.73 (d, J = 8.5 Hz, 2H), 7.43 (d, J = 8.2 Hz, 2H), 7.12759(m, 4H), 4.49 (d, J = 5.7 Hz, 2H), 3.84, (s, 3H), 2.57−2.50 (m, 2H),7601.16 (t, J = 7.4 Hz, 3H ), ppm; LC-MS: m/z = 438.2 [M + H]+.7614-Chloro-3-ethyl-1-methyl-N-(4-(3,3,3-trifluoropropoxy)-762benzyl)-1H-pyrazole-5-carboxamide (18). General procedure A2763was followed, starting from 3,3,3-trifluoropropan-1-ol and 4-764fluorobenzonitrile to give 4-(3,3,3-trifluoropropoxy)benzonitrile,765which was then reduced according to General procedure B2 to give766the corresponding benzylamine that was subsequently coupled to 4-767chloro-3-ethyl-1-methyl-1H-pyrazole-5-carboxylic acid according to768General procedure C3 to give the title compound as an off-white solid769(50 mg, 14%). 1H NMR (400 MHz, DMSO-d6): δ = 8.86 (t, J = 5.7770Hz, 1H), 7.27 (d, J = 8.5 Hz, 2H), 6.93 (d, J = 8.6 Hz, 2H), 4.39 (d, J771= 6.0 Hz, 2H), 4.18 (t, J = 5.9 Hz, 2H), 3.82 (s, 3H), 2.81−2.72 (m,7722H), 2.50−2.56 (m, 2H), 1.15 (t, J = 7.5 Hz, 3H) ppm; LC-MS: m/z773= 390.1 [M + H]+.7745- (4- ( (4-Chloro-3-ethyl -1-methyl -1H -pyrazole-5-775carboxamido)methyl)phenoxy)-2-methylpyridine 1-oxide776(19). Compound 12 (0.3 mmol) was dissolved in DCM, and m-777CPBA was then added. The reaction mixture was stirred at room778temperature for 2 h. DCM was then removed in vacuo and EtOAc779was added to the residue. The organic layer was washed with780saturated NaHCO3 and brine, dried (MgSO4), and concentrated in781vacuo to give crude product, which was then purified by column782chromatography (5% MeOH/DCM) to give the title compound as a783yellow oil (13%). 1H NMR (400 MHz, CDCl3) δ = 8.07 (d, J = 2.0784Hz, 1H), 7.38 (d, J = 8.6 Hz, 2H), 7.18 (d, J = 8.7 Hz, 1H), 7.08 (s,785br, 1H), 7.05−7.02 (m, 2H), 6.91 (d, J = 8.7 Hz, 1H), 4.63 (d, J = 5.9786Hz, 2H), 4.14 (s, 3H), 2.63 (q, J = 7.6 Hz, 2H), 2.49 (s, 3H), 1.23 (t,787J = 7.5 Hz, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ = 158.7, 154.7,788154.6, 149.7, 144.2, 134.7, 131.2, 130.9, 129.7, 126.1, 120.0, 117.1,789107.8, 42.9, 40.8, 19.3, 17.2, 12.9 ppm; LC-MS: m/z = 400.8 [M +790H]+.791N-(4-(Benzo[d]oxazol-5-yloxy)benzyl)-4-chloro-3-ethyl-1-792methyl-1H-pyrazole-5-carboxamide (20). General procedure B3793was followed, starting from 74 to give the desired benzylamine, which794was subsequently coupled to 4-chloro-3-ethyl-1-methyl-1H-pyrazole-7955-carboxylic acid according to General procedure C3 to afford the title796compound as a brown solid (10%). 1H NMR (400 MHz, CDCl3): δ =7978.09 (t, J = 4.8 Hz, 1H), 7.55 (d, J = 8.7 Hz, 1H), 7.40 (s, 1H), 7.32798(d, J = 7.8 Hz, 2H), 7.12−7.10 (m, 1H), 6.99−6.97 (m, 3H), 4.60 (d,799J = 4.8 Hz, 2H), 4.13 (s, 3H), 2.63−2.61 (m, 2H), 1.23 (t, J = 7.7 Hz,8003H) ppm; LCMS: m/z = 411.2 [M + H]+.8014- (4- ( (4-Chloro-3-ethyl -1-methyl -1H -pyrazole-5-802carboxamido)methyl)phenoxy)benzoic acid (21). Title com-803pound was prepared according to General procedure D1, starting804from 76 to give a white solid (17%). 1H NMR (400 MHz, DMSO-805d6): δ = 12.8 (s. br, 1H), 8.95 (t, J = 5.9 Hz, 1H), 7.93 (d, J = 8.6 Hz,8062H), 7.42 (d, J = 8.3 Hz, 2H), 7.10 (d, J = 8.4 Hz, 2H), 7.01 (d, J =8078.6 Hz, 2H), 4.49 (d, J = 5.9 Hz, 2H), 3.84 (s, 3H), 2.57−2.49 (m,8082H), 1.16 (t, J = 7.6 Hz, 3H) ppm; LC-MS: m/z = 414.1 [M + H]+.809N-(4-(4-(Azidomethyl)phenoxy)benzyl)-4-chloro-3-ethyl-1-810methyl-1H-pyrazole-5-carboxamide (22). General procedure C1811was followed, starting from 4-hydroxybenzylamine and 4-chloro-3-812ethyl-1-methyl-1H-pyrazole-5-carboxylic acid to give 4-chloro-3-ethyl-813N-(4-hydroxybenzyl)-1-methyl-1H-pyrazole-5-carboxamide, which814was then coupled to (4-iodophenyl)methanol according to General815procedure E1 to give 4-chloro-3-ethyl-N-(4-(4-(hydroxymethyl)-816phenoxy)benzyl)-1-methyl-1H-pyrazole-5-carboxamide. This result-817ing product (1.0 equiv) was reacted with triphenylphosphine (1.1818equiv) and CBr4 (1.1 equiv) in DCM at room temperature for 2 h.819DCM was then removed in vacuo and DMF was added to the residue,820followed by NaN3 (5.5 equiv). The reaction mixture was stirred at



K


821 room temperature for 2 h. Upon completion, the reaction was diluted822 with EtOAc, washed with water and brine, and the organic layer was823 dried over MgSO4. Solvent was removed in vacuo to afford crude824 product, which was purified by column chromatography (5−20%825 EtOAc/petroleum benzine) to give the title compound as a colorless826 oil (24%). 1H NMR (401 MHz, CDCl3) δ = 7.33 (d, J = 8.6 Hz, 2H),827 7.28 (d, J = 8.6 Hz, 2H), 7.01 (d, J = 8.0 Hz, 5H), 4.62 (d, J = 5.8 Hz,828 2H), 4.31 (s, 2H), 4.15 (s, 3H), 2.63 (q, J = 7.6 Hz, 2H), 1.24 (t, J =829 7.6 Hz, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ = 183.3, 158.6,830 156.5, 149.7, 132.8, 131.0, 130.4, 130.0, 129.3, 119.5, 119.1, 107.0,831 54.4, 43.0, 40.8, 19.3, 12.9 ppm; LC-MS: m/z = 424.8 [M + H]+.832 N-(4-(4-Chlorophenoxy)benzyl)-3-ethyl-1-methyl-1H-pyra-833 zole-5-carboxamide (23). Title compound was prepared according834 to General procedure C1, starting from (4-(4-chlorophenoxy)-835 phenyl)methanamine HCl and 3-ethyl-1-methyl-1H-pyrazole-5-car-836 boxylic acid to give a yellow oil (58%). 1H NMR (400 MHz, CDCl3)837 δ = 7.32−7.25 (m, 4H), 6.99−6.90 (m, 4H), 6.39 (s, br, 1H), 6.31 (s,838 1H), 4.54 (d, J = 5.8 Hz, 2H), 4.12 (s, 3H), 2.61 (q, J = 7.6 Hz, 2H),839 1.21 (t, J = 7.6 Hz, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ =840 160.1, 156.6, 155.8, 153.0, 135.5, 133.2, 129.8, 129.5, 128.5, 120.2,841 119.2, 104.2, 42.9, 39.0, 21.3, 13.9 ppm; LC-MS: m/z = 369.9 [M +842 H]+.843 4-Chloro-1-methyl-N-(4-((6-methylpyridin-3-yl)oxy)benzyl)-844 1H-pyrazole-5-carboxamide (24). General procedure A1 was845 followed, starting from 6-methylpyridin-3-ol and 4-fluorobenzalde-846 hyde to give 4-((6-methylpyridin-3-yl)oxy)benzaldehyde, which was847 converted to the corresponding benzylamine according to General848 procedure F, and then subsequently coupled to 4-chloro-1-methyl-849 1H-pyrazole-5-carboxylic acid according to General procedure C1 to850 give the title compound as a yellow oil (35%).1H NMR (400 MHz,851 CDCl3) δ = 8.26 (d, J = 2.8 Hz, 1H), 7.43 (d, J = 4.0 Hz, 1H), 7.37−852 7.32 (m, 3H), 7.20 (d, J = 8.5 Hz, 1H), 7.06−6.96 (m, 3H), 4.62 (d, J853 = 5.8 Hz, 2H), 4.19 (s, 3H), 2.61 (s, 3H) ppm; 13C NMR (101 MHz,854 CDCl3) δ = 166.6, 158.4, 156.1, 155.7, 136.8, 133.4, 130.9, 129.6,855 129.0, 128.2, 124.6, 119.0, 109.7, 43.0, 41.3, 22.9 ppm; LC-MS: m/z856 = 356.8 [M + H]+.857 4-Chloro-N-(4-(4-chlorophenoxy)benzyl)-1-methyl-1H-pyra-858 zole-5-carboxamide (25). Title compound was prepared according859 to General procedure C1, starting from (4-(4-chlorophenoxy)-860 phenyl)methanamine HCl and 4-chloro-1-methyl-1H-pyrazole-5-861 carboxylic acid to give a white solid (36%). 1H NMR (400 MHz,862 CDCl3) δ = 7.44 (d, J = 1.3 Hz, 1H), 7.35−7.24 (m, 4H), 7.06−6.89863 (m, 5H), 4.61 (d, J = 5.7 Hz, 2H), 4.19 (s, 3H) ppm; 13C NMR (101864 MHz, CDCl3) δ = 158.4, 156.6, 155.8, 136.7, 132.7, 130.9, 129.9,865 129.3, 128.5, 120.2, 119.2, 109.6, 43.0, 41.2 ppm; LC-MS: m/z =866 375.7 [M + H]+.867 4-Chloro-1-methyl-N-(4-((2-methylpyrimidin-5-yl)oxy)-868 benzyl)-1H-pyrazole-5-carboxamide (26). General procedure A1869 was followed, starting from 2-methylpyrimidin-5-ol and 4-fluoroben-870 zonitrile to give 4-((2-methylpyrimidin-5-yl)oxy)benzonitrile, which871 was converted to the corresponding benzylamine according to872 General procedure B4, and then subsequently coupled to 4-chloro-873 1-methyl-1H-pyrazole-5-carboxylic acid according to General proce-874 dure C1 to give the title compound as a yellow oil (16%). 1H NMR875 (400 MHz, CDCl3) δ = 8.39 (s, 2H), 7.44 (s, 1H), 7.36 (d, J = 8.4876 Hz, 2H), 7.00 (d, J = 8.2 Hz, 3H), 4.63 (d, J = 5.8 Hz, 2H), 4.18 (s,877 3H), 2.72 (s, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ = 163.1,878 158.4, 155.8, 149.7, 147.9, 136.8, 133.7, 130.8, 129.6, 118.8, 109.7,879 42.9, 41.3, 25.2 ppm; LC-MS: m/z = 357.8 [M + H]+.880 4-Chloro-N-(4-((6-chloropyridin-3-yl)oxy)benzyl)-1-methyl-881 1H-pyrazole-5-carboxamide (27). General procedure A1 was882 followed, starting from 6-chloropyridin-3-ol and 4-fluorobenzonitrile883 to give 4-((6-chloropyridin-3-yl)oxy)benzonitrile, which was con-884 verted to the corresponding benzylamine according to General885 procedure B4, and then subsequently coupled to 4-chloro-1-methyl-886 1H-pyrazole-5-carboxylic acid according to General procedure C1 to887 give the title compound as a yellow oil (23%). 1H NMR (400 MHz,888 MeOD) δ = 8.08 (dd, J = 2.5, 1.1 Hz, 1H), 7.50−7.38 (m, 6H), 7.09−889 7.03 (m, 2H), 4.58 (s, 2H), 3.99 (s, 3H) ppm; 13C NMR (101 MHz,890 CDCl3) δ = 158.4, 155.9, 152.2, 138.3, 136.7, 136.7, 133.5, 130.9,

891129.6, 128.4, 124.8, 119.0, 109.6, 42.9, 41.2, 22.6 ppm; LC-MS: m/z892= 376.8 [M + H]+.8934-Fluoro-1-methyl-N-(4-((6-methylpyridin-3-yl)oxy)benzyl)-8941H-pyrazole-5-carboxamide (28). General procedure A1 was895followed, starting from 6-methylpyridin-3-ol and 4-fluorobenzalde-896hyde to give 4-((6-methylpyridin-3-yl)oxy)benzaldehyde, which was897converted to the corresponding benzylamine according to General898procedure F, and then subsequently coupled to 4-fluoro-1-methyl-1H-899pyrazole-5-carboxylic acid according to General procedure C1 to give900the title compound as a yellow oil (48%). 1H NMR (400 MHz,901CDCl3) δ = 8.27 (d, J = 2.6 Hz, 1H), 7.33−7.27 (m, 3H), 7.21 (dd, J902= 8.5, 2.8 Hz, 1H), 7.11 (d, J = 8.5 Hz, 1H), 6.97−6.93 (m, 2H), 6.56903(s, br, 1H), 4.57 (d, J = 5.8 Hz, 2H), 4.14 (d, J = 0.9 Hz, 3H), 2.52 (s,9043H) ppm; 13C NMR (101 MHz, CDCl3) δ = 157.8, 156.7, 153.4,905151.3, 150.0, 147.5, 140.8, 132.9, 129.4, 126.9, 124.39, 123.8, 118.6,90642.6, 40.8, 23.6 ppm; LC-MS: m/z = 340.9 [M + H]+.907N-(4-(4-Chlorophenoxy)benzyl)-4-fluoro-1-methyl-1H-pyra-908zole-5-carboxamide (29). Title compound was prepared according909to General procedure C1, starting from (4-(4-chlorophenoxy)-910phenyl)methanamine HCl and 4-fluoro-1-methyl-1H-pyrazole-5-911carboxylic acid to give a white solid (17%). 1H NMR (400 MHz,912CDCl3) δ = 7.35−7.25 (m, 5H), 7.00−6.91 (m, 4H), 6.51 (s, br, 1H),9134.59 (d, J = 5.8 Hz, 2H), 4.17 (d, J = 1.0 Hz, 3H) ppm; 13C NMR914(101 MHz, CDCl3) δ = 157.8, 156.6, 155.8, 150.1, 147.6, 132.9,915129.9, 129.4, 128.6, 124.4, 120.2, 119.2, 42.7, 40.9 ppm; LC-MS: m/z916= 357.9 [M − H]−.9174-Fluoro-1-methyl-N-(4-((2-methylpyrimidin-5-yl)oxy)-918benzyl)-1H-pyrazole-5-carboxamide (30). General procedure A1919was followed, starting from 2-methylpyrimidin-5-ol and 4-fluoroben-920zonitrile to give 4-((2-methylpyrimidin-5-yl)oxy)benzonitrile, which921was converted to the corresponding benzylamine according to922General procedure B4, and then subsequently coupled to 4-fluoro-9231-methyl-1H-pyrazole-5-carboxylic acid according to General proce-924dure C1 to give the title compound as a yellow oil (18%). 1H NMR925(400 MHz, MeOD) δ = 8.43 (s, 2H), 7.45−7.41 (m, 3H), 7.11−7.06926(m, 2H), 4.56 (s, 2H), 4.03 (d, J = 0.8 Hz, 3H), 2.66 (s, 3H) ppm;927

13C NMR (101 MHz, MeOD) δ = 163.3, 156.4, 151.9, 151.1, 148.4,928148.4, 136.6, 130.6, 125.6, 120.10, 120.0, 43.3, 40.3, 24.6 ppm; LC-929MS: m/z = 341.9 [M + H]+.930N-(4-((6-Chloropyridin-3-yl)oxy)benzyl)-4-fluoro-1-methyl-9311H-pyrazole-5-carboxamide (31). General procedure A1 was932followed, starting from 6-chloropyridin-3-ol and 4-fluorobenzonitrile933to give 4-((6-chloropyridin-3-yl)oxy)benzonitrile, which was con-934verted to the corresponding benzylamine according to General935procedure B4, and then subsequently coupled to 4-fluoro-1-methyl-9361H-pyrazole-5-carboxylic acid according to General procedure C1 to937give the title compound as a yellow oil (7%). 1H NMR (400 MHz,938MeOD) δ = 8.09 (dd, J = 2.4, 1.1 Hz, 1H), 7.46−7.39 (m, 5H), 7.09−9397.04 (m, 2H), 4.56 (d, J = 4.2 Hz, 2H), 4.03 (d, J = 0.8 Hz, 3H) ppm;940

13C NMR (101 MHz, MeOD) δ = 186.5, 185.2, 178.6, 175.6, 173.2,941171.0, 166.4, 160.5, 160.1, 156.3, 155.6, 150.4, 73.3, 70.3 ppm; LC-942MS: m/z = 360.8 [M + H]+.9434-Chloro-1,3-dimethyl-N-(4-((6-methylpyridin-3-yl)oxy)-944benzyl)-1H-pyrazole-5-carboxamide (32). General procedure A1945was followed, starting from 6-methylpyridin-3-ol and 4-fluorobenzal-946dehyde to give 4-((6-methylpyridin-3-yl)oxy)benzaldehyde, which947was converted to the corresponding benzylamine according to948General procedure F, and then subsequently coupled to 4-chloro-9491,3-dimethyl-1H-pyrazole-5-carboxylic acid according to General950procedure C1 to give the title compound as a yellow oil (40%). 1H951NMR (400 MHz, CDCl3) δ = 8.28 (d, J = 2.8 Hz, 1H), 7.33−7.29952(m, 2H), 7.21 (dd, J = 8.4, 2.9 Hz, 1H), 7.11 (d, J = 8.5 Hz, 1H), 7.03953(s, br, 1H), 6.98−6.93 (m, 2H), 4.59 (d, J = 5.8 Hz, 2H), 4.12 (s,9543H), 2.53 (s, 3H), 2.22 (s, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ955= 158.5, 156.8, 153.5, 151.3, 144.6, 141.0, 132.8, 131.0, 129.3, 126.8,956123.8, 118.6, 108.5, 42.9, 40.7, 23.7, 11.1 ppm; LC-MS: m/z = 370.8957[M + H]+.9584-Chloro-N-(4-(4-chlorophenoxy)benzyl)-1,3-dimethyl-1H-959pyrazole-5-carboxamide (33). Title compound was prepared960according to General procedure C1, starting from (4-(4-



L


961 chlorophenoxy)phenyl)methanamine HCl and 4-chloro-1,3-dimethyl-962 1H-pyrazole-5-carboxylic acid to give a yellow solid (37%). 1H NMR963 (400 MHz, CDCl3) δ = 7.34−7.26 (m, 4H), 7.04 (s, br, 1H), 7.00−964 6.90 (m, 4H), 4.61 (d, J = 5.8 Hz, 2H), 4.13 (s, 3H), 2.23 (s, 3H)965 ppm; 13C NMR (101 MHz, CDCl3) δ = 158.5, 156.6, 155.8, 144.6,966 132.8, 131.0, 129.8, 129.3, 128.5, 120.2, 119.2, 108.4, 42.9, 40.7, 11.1967 ppm; LC-MS: m/z = 389.8 [M + H]+.968 4-Chloro-N-(4-((6-chloropyridin-3-yl)oxy)benzyl)-1,3-di-969 methyl-1H-pyrazole-5-carboxamide (34). General procedure A1970 was followed, starting from 6-chloropyridin-3-ol and 4-fluorobenzoni-971 trile to give 4-((6-chloropyridin-3-yl)oxy)benzonitrile, which was972 converted to the corresponding benzylamine according to General973 procedure B4, and then subsequently coupled to 4-chloro-1,3-974 dimethyl-1H-pyrazole-5-carboxylic acid according to General proce-975 dure C1 to give the title compound as a yellow oil (10%). 1H NMR976 (400 MHz, MeOD) δ = 8.09 (d, J = 1.1 Hz, 1H), 7.48−7.41 (m, 4H),977 7.07 (dd, J = 8.5, 1.7 Hz, 2H), 4.58 (s, 2H), 3.93 (d, J = 1.7 Hz, 3H),978 2.20 (d, J = 1.7 Hz, 3H) ppm; LC-MS: m/z = 390.8 [M + H]+.979 4-Fluoro-1,3-dimethyl-N-(4-((6-methylpyridin-3-yl)oxy)-980 benzyl)-1H-pyrazole-5-carboxamide (35). General procedure A1981 was followed, starting from 6-methylpyridin-3-ol and 4-fluorobenzal-982 dehyde to give 4-((6-methylpyridin-3-yl)oxy)benzaldehyde, which983 was converted to the corresponding benzylamine according to984 General procedure F, and then subsequently coupled to 4-fluoro-985 1,3-dimethyl-1H-pyrazole-5-carboxylic acid according to General986 procedure C1 to give the title compound as a yellow oil (12%). 1H987 NMR (400 MHz, CDCl3) δ = 8.29 (d, J = 2.7 Hz, 1H), 7.32−7.28988 (m, 2H), 7.23−7.19 (m, 1H), 7.12 (d, J = 8.5 Hz, 1H), 6.99−6.94 (m,989 2H), 6.52 (s, br, 1H), 4.58 (d, J = 5.8 Hz, 2H), 4.10 (d, J = 0.8 Hz,990 3H), 2.54 (s, 3H), 2.21 (s, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ991 = 158.0, 156.8, 153.5, 151.3, 148.1, 145.6, 141.0, 133.0, 132.8 (d, J =992 11.7 Hz), 129.4, 126.8, 123.8, 118.7, 42.6, 40.2, 23.7, 9.7 (d, J = 3.2993 Hz) ppm; LC-MS: m/z = 354.9 [M + H]+.994 N-(4-(4-Chlorophenoxy)benzyl)-4-fluoro-1,3-dimethyl-1H-995 pyrazole-5-carboxamide (36). Title compound was prepared996 according to General procedure C1, starting from (4-(4-997 chlorophenoxy)phenyl)methanamine HCl and 4-fluoro-1,3-dimeth-998 yl-1H-pyrazole-5-carboxylic acid to give a colorless oil (30%). 1H999 NMR (400 MHz, CDCl3) δ = 7.34−7.25 (m, 4H), 7.01−6.90 (m,1000 4H), 6.52 (s, br, 1H), 4.59 (d, J = 5.8 Hz, 2H), 4.10 (d, J = 0.8 Hz,1001 3H), 2.22 (s, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ = 157.9,1002 156.4, 155.7, 148.0, 145.5, 132.9, 132.63, 129.7, 129.2, 128.4, 120.1,1003 119.1, 42.5, 40.1, 9.5 (d, J = 3.2 Hz) ppm; LC-MS: m/z = 373.8 [M +1004 H]+.1005 N-(4-((6-Chloropyridin-3-yl)oxy)benzyl)-4-fluoro-1,3-di-1006 methyl-1H-pyrazole-5-carboxamide (37). General procedure A11007 was followed, starting from 6-chloropyridin-3-ol and 4-fluorobenzoni-1008 trile to give 4-((6-chloropyridin-3-yl)oxy)benzonitrile, which was1009 converted to the corresponding benzylamine according to General1010 procedure B4, and then subsequently coupled to 4-fluoro-1,3-1011 dimethyl-1H-pyrazole-5-carboxylic acid according to General proce-1012 dure C1 to give the title compound as a colorless oil (38%). 1H NMR1013 (400 MHz, CDCl3) δ = 8.11−8.07 (m, 1H), 7.33−7.28 (m, 2H), 7.231014 (d, J = 1.9 Hz, 2H), 6.99−6.93 (m, 2H), 6.60 (d, J = 6.0 Hz, 1H),1015 4.56 (d, J = 5.9 Hz, 2H), 4.05 (d, J = 0.9 Hz, 3H), 2.17 (s, 3H) ppm;1016

13C NMR (101 MHz, CDCl3) δ = 158.0, 157.9, 155.5, 153.1, 148.0,1017 145.5, 144.9, 140.5, 134.1, 132.6 (d, J = 11.7 Hz), 129.5, 128.57,1018 124.8, 119.2, 42.4, 40.1, 9.5 (d, J = 3.1 Hz) ppm; LC-MS: m/z =1019 374.8 [M + H]+.1020 4-Chloro-3-ethyl-1-methyl-N-((5-(p-tolyloxy)pyridin-2-yl)-1021 methyl)-1H-pyrazole-5-carboxamide (38). Title compound was1022 prepared according to General procedure C5, starting from 78 and 4-1023 chloro-3-ethyl-1-methyl-1H-pyrazole-5-carboxylic acid to give a white1024 solid (19%). 1H NMR (400 MHz, DMSO d6): δ 8.94 (t, J = 5.6 Hz,1025 1H), 8.28 (s, 1H), 7.39 (s, 2H), 7.22 (d, J = 8.0 Hz, 2H), 6.96 (d, J =1026 8.4 Hz, 2H), 4.56 (d, J = 5.6 Hz, 2H), 3.87 (s, 3H), 2.58−2.50 (m,1027 2H), 2.30 (s, 3H), 1.17 (t, J = 7.5 Hz, 3H) ppm; LC-MS: m/z = 385.11028 [M + H]+.1029 4-Chloro-3-ethyl-N-(2-fluoro-4-(p-tolyloxy)benzyl)-1-meth-1030 yl-1H-pyrazole-5-carboxamide (39). Title compound was pre-

1031pared according to General procedure C2, starting from 80 and 4-1032chloro-3-ethyl-1-methyl-1H-pyrazole-5-carboxylic acid to give a color-1033less gum (62%). 1H NMR (400 MHz, DMSO-d6): δ 8.88 (t, J = 5.61034Hz, 1H), 7.39 (t, J = 8.6 Hz, 1H), 7.22 (d, J = 8.2 Hz, 2H), 6.96 (d, J1035= 8.4 Hz, 2H), 6.84−6.80 (m, 1H), 6.77 (dd, J = 8.4, 2.36 Hz, 1H),10364.46 (d, J = 5.7 Hz, 2H), 3.83 (s, 3H), 2.56-2.50 (m, 2H), 2.30 (s,10373H), 1.16 (t, J = 7.5 Hz, 3H) ppm; LC-MS: m/z = 402.1 [M + H]+.10384-Chloro-3-ethyl-1-methyl-N-((6-(p-tolyloxy)pyridin-3-yl)-1039methyl)-1H-pyrazole-5-carboxamide (40). Title compound was1040prepared according to General procedure C2, starting from 82 and 4-1041chloro-3-ethyl-1-methyl-1H-pyrazole-5-carboxylic acid to give an off-1042white solid (23%). 1H NMR (400 MHz, DMSO-d6): δ 8.93 (t, J = 5.71043Hz, 1H), 8.10 (s, 1H), 7.80 (d, J = 6.4 Hz, 1H), 7.20 (d, J = 8.1 Hz,10442H), 6.99−6.96 (m, 3H), 4.43 (d, J = 5.7 Hz, 2H), 3.83 (s, 3H),10452.54−2.50 (m, 2H), 2.31 (s, 3H), 1.16 (t, J = 7.5 Hz, 3H) ppm; LC-1046MS: m/z = 385.1 [M + H]+.10474-Chloro-3-ethyl-N-(3-fluoro-4-(p-tolyloxy)benzyl)-1-meth-1048yl-1H-pyrazole-5-carboxamide (41). Title compound was pre-1049pared according to General procedure C2, starting from 84 and 4-1050chloro-3-ethyl-1-methyl-1H-pyrazole-5-carboxylic acid to give an off-1051white solid (44%). 1H NMR (400 MHz, DMSO-d6): δ 8.96 (s. br,10521H), 7.35−7.32 (m, 1H), 7.18−7.07 (m, 4H), 6.86 (d, J = 8.0 Hz,10532H), 4.48 (d, J = 5.3 Hz, 2H), 3.84 (s, 3H), 2.56−2.50 (m, 2H), 2.271054(s, 3H), 1.17 (t, J = 7.3 Hz, 3H) ppm; LC-MS: m/z = 402.2 [M +1055H]+.10564-Chloro-3-ethyl-N-(2-fluoro-4-((6-methylpyridin-3-yl)oxy)-1057benzyl)-1-methyl-1H-pyrazole-5-carboxamide (42). General1058procedure C1 was followed, starting from (4-bromo-2-fluorophenyl)-1059methanamine and 4-chloro-3-ethyl-1-methyl-1H-pyrazole-5-carbox-1060ylic acid to give N-(4-bromo-2-fluorobenzyl)-4-chloro-3-ethyl-1-1061methyl-1H-pyrazole-5-carboxamide, which was then coupled to 6-1062methylpyridin-3-ol according to General procedure E1 to give the title1063compound as a colorless oil (37%). 1H NMR (400 MHz, CDCl3) δ =10648.27 (s, 1H), 7.34 (t, J = 8.4 Hz, 1H), 7.23 (dd, J = 8.5, 2.3 Hz, 1H),10657.13 (s, br, 2H), 6.74−6.66 (m, 2H), 4.60 (d, J = 5.9 Hz, 2H), 4.09 (s,10663H), 2.59 (q, J = 7.6 Hz, 2H), 2.52 (s, 3H), 1.20 (t, J = 7.6 Hz, 3H)1067ppm; 13C NMR (101 MHz, CDCl3) δ = 162.8, 160.3, 158.6, 158.2 (d,1068J = 10.7 Hz), 154.3, 149.6, 141.2, 131.1 (d, J = 5.8 Hz), 130.9, 127.4,1069119.7 (d, J = 15.2 Hz), 113.6 (d, J = 3.4 Hz), 107.7, 105.9, 105.69,107040.6, 37.2 (d, J = 3.2 Hz), 23.7, 19.2, 12.8 ppm; LC-MS: m/z = 402.81071[M + H]+.10724-Chloro-N-(4-(4-chlorophenoxy)-2-fluorobenzyl)-3-ethyl-10731-methyl-1H-pyrazole-5-carboxamide (43). Title compound was1074prepared according to General procedure C2, starting from 86 and 4-1075chloro-3-ethyl-1-methyl-1H-pyrazole-5-carboxylic acid to give an off-1076white solid (24%). 1H NMR (400 MHz, DMSO-d6): δ 8.90 (s, br,10771H), 7.47−7.41 (m, 3H), 7.08 (d, J = 8.7 Hz, 2H), 6.95 (d, J = 11.31078Hz, 1H), 6.86 (d, J = 7.4 Hz, 1H), 4.48 (d, J = 5.1 Hz, 2H), 3.83 (s,10793H), 2.55−2.50 (m, 2H), 1.16 (t, J = 7.4 Hz, 3H) ppm; LC-MS: m/z1080= 422.0 [M + H]+.10814-Chloro-3-ethyl-N-(3-fluoro-4-((6-methylpyridin-3-yl)oxy)-1082benzyl)-1-methyl-1H-pyrazole-5-carboxamide (44). General1083procedure A1 was followed, starting from 6-methylpyridin-3-ol and10843,4-difluorobenzaldehyde to give 3-fluoro-4-((6-methylpyridin-3-yl)-1085oxy)benzaldehyde, which was converted to the corresponding1086benzylamine according to General procedure F, and then1087subsequently coupled to 4-chloro-3-ethyl-1-methyl-1H-pyrazole-5-1088carboxylic acid according to General procedure C1 to give the title1089compound as a yellow oil (28%). 1H NMR (400 MHz, MeOD) δ10908.12 (d, J = 2.7 Hz, 1H), 7.34−7.21 (m, 4H), 7.14 (t, J = 8.3 Hz, 1H),10914.58 (s, 2H), 3.94 (s, 3H), 2.62 (q, J = 7.6 Hz, 2H), 2.49 (s, 3H), 1.221092(t, J = 7.6 Hz, 3H) ppm; 13C NMR (101 MHz, MeOD) δ = 160.8,1093156.6, 154.1, 153.9 (d, J = 6.7 Hz), 151.0, 143.1 (d, J = 11.8 Hz),1094138.8, 138.3 (d, J = 6.2 Hz), 134.7, 126.5, 125.6, 125.3 (d, J = 3.51095Hz), 123.2 (d, J = 0.9 Hz), 117.4 (d, J = 19.0 Hz), 108.9, 43.3, 39.4,109622.9, 19.9, 13.2 ppm; LC-MS: m/z = 402.8 [M + H]+.10974-Chloro-N-(4-(4-chlorophenoxy)-3-fluorobenzyl)-3-ethyl-10981-methyl-1H-pyrazole-5-carboxamide (45). General procedure1099B2 was followed, starting from 87 to give the desired benzylamine,1100which was subsequently coupled to 4-chloro-3-ethyl-1-methyl-1H-



M


1101 pyrazole-5-carboxylic acid according to General procedure C2 to1102 afford the title compound as an off-white solid (28%). 1H NMR (4001103 MHz, DMSO-d6): δ 8.97 (s. br, 1H), 7.43−7.35 (m, 3H), 7.22 (s. br,1104 2H), 6.98 (d, J = 8.5 Hz, 2H), 4.49 (d, J = 5.3 Hz, 2H), 3.85 (s, 3H),1105 2.56−2.50 (m, 2H), 1.17 (t, J = 7.4 Hz, 3H) ppm; LC-MS: m/z =1106 422.1 [M + H]+.1107 4-Chloro-N-(3,5-difluoro-4-((6-methylpyridin-3-yl)oxy)-1108 benzyl)-3-ethyl-1-methyl-1H-pyrazole-5-carboxamide (46).1109 General procedure A1 was followed, starting from 6-methylpyridin-1110 3-ol and 3,4,5-trifluorobenzaldehyde to give 3,5-difluoro-4-((6-1111 methylpyridin-3-yl)oxy)benzaldehyde, which was converted to the1112 corresponding benzylamine according to General procedure F, and1113 then subsequently coupled to 4-chloro-3-ethyl-1-methyl-1H-pyrazole-1114 5-carboxylic acid according to General procedure C1 to give the title1115 compound as a white solid (31%). 1H NMR (400 MHz, CDCl3) δ =1116 8.25 (d, J = 2.8 Hz, 1H), 7.21−7.12 (m, 2H), 7.09 (d, J = 8.5 Hz,1117 1H), 7.05−6.98 (m, 2H), 4.62 (d, J = 6.1 Hz, 2H), 4.14 (s, 3H), 2.641118 (q, J = 7.6 Hz, 2H), 2.52 (s, 3H), 1.24 (t, J = 7.6 Hz, 3H) ppm; 13C1119 NMR (101 MHz, CDCl3) δ = 158.9, 157.30 (d, J = 4.7 Hz), 154.7 (d,1120 J = 4.9 Hz), 152.9, 152.4, 149.8, 137.2, 136.6, 130.6, 123.6 (d, J = 27.71121 Hz), 111.7 (d, J = 5.6 Hz), 111.5 (d, J = 5.6 Hz), 108.0, 42.4, 40.9,1122 23.4, 19.3, 12.9 ppm; LC-MS: m/z = 420.8 [M + H]+.1123 4-Chloro-3-ethyl-1-methyl-N-(4-(4-methylbenzyl)benzyl)-1124 1H-pyrazole-5-carboxamide (47). General procedure B1 was1125 followed, starting from 88 to give the corresponding benzylamine,1126 which was subsequently coupled to 4-chloro-3-ethyl-1-methyl-1H-1127 pyrazole-5-carboxylic acid according to General procedure C1 to give1128 the title compound as a white solid (45%). 1H NMR (400 MHz,1129 CDCl3) δ = 7.28−7.24 (m, 2H), 7.18 (d, J = 8.1 Hz, 2H), 7.11−7.051130 (m, 4H), 6.99 (s, br, 1H), 4.60 (d, J = 5.7 Hz, 2H), 4.14 (s, 3H), 3.931131 (s, 2H), 2.62 (q, J = 7.6 Hz, 2H), 2.31 (s, 3H), 1.23 (t, J = 7.6 Hz,1132 3fH) ppm; 13C NMR (101 MHz, CDCl3) δ = 158.6, 149.7, 141.1,1133 137.9, 135.8, 135.2, 129.4, 129.3, 128.9, 128.3, 127.9, 107.7, 43.3,1134 41.3, 40.8, 21.1, 19.3, 12.9 ppm; LC-MS: m/z = 381.9 [M + H]+.1135 4-Chloro-3-ethyl-1-methyl-N-(4-(4-methylbenzoyl)benzyl)-1136 1H-pyrazole-5-carboxamide (48). General procedure C1 was1137 followed, starting from methyl 4-(aminomethyl)benzoate HCl and 4-1138 chloro-3-ethyl-1-methyl-1H-pyrazole-5-carboxylic acid to give methyl1139 4-((4-chloro-3-ethyl-1-methyl-1H-pyrazole-5-carboxamido)methyl)-1140 benzoate, which was hydrolyzed according to General procedure D31141 to give the corresponding carboxylic acid that subsequently turned1142 into a Weinreb amide according to General procedure C1. The1143 resulting Weinreb amide was subjected to General procedure H1,1144 reacting with p-tolylmagnesium bromide to give the title compound as1145 a white solid (78%). 1H NMR (400 MHz, CDCl3) δ = 7.73−7.68 (m,1146 2H), 7.63 (d, J = 8.2 Hz, 2H), 7.38 (d, J = 8.2 Hz, 2H), 7.22−7.181147 (m, 2H), 7.09 (s, br, 1H), 4.65 (d, J = 5.9 Hz, 2H), 4.07 (s, 3H), 2.571148 (q, J = 7.6 Hz, 2H), 2.36 (s, 3H), 1.17 (t, J = 7.6 Hz, 3H) ppm; 13C1149 NMR (101 MHz, CDCl3) δ = 196.0, 158.8, 149.7, 143.4, 142.0,1150 137.4, 134.9, 130.9, 130.6, 130.38, 129.1, 127.3, 107.8, 43.2, 40.8,1151 21.7, 19.3, 12.9 ppm; LC-MS: m/z = 395.9 [M + H]+.1152 4-Chloro-3-ethyl-1-methyl-N-(4-(p-tolylthio)benzyl)-1H-pyr-1153 azole-5-carboxamide (49). General procedure B1 was followed,1154 starting from 89 to give the corresponding benzylamine, which was1155 subsequently coupled to 4-chloro-3-ethyl-1-methyl-1H-pyrazole-5-1156 carboxylic acid to give the title compound as a yellow solid (79%).1157

1H NMR (400 MHz, CDCl3) δ = 7.33−7.28 (m, 2H), 7.25 (d, J = 7.71158 Hz, 4H), 7.14 (d, J = 7.9 Hz, 2H), 7.01 (s, br, 1H), 4.59 (d, J = 5.81159 Hz, 2H), 4.14 (s, 3H), 2.63 (q, J = 7.6 Hz, 2H), 2.35 (s, 3H), 1.23 (t,1160 J = 7.6 Hz, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ = 158.6, 149.7,1161 138.0, 136.9, 135.8, 132.6, 131.0, 131.0, 130.2, 130.0, 128.4, 107.7,1162 43.1, 40.8, 21.2, 19.3, 12.9 ppm; LC-MS: m/z = 399.8 [M + H]+.1163 4-Chloro-3-ethyl-1-methyl-N-(4-(methyl(p-tolyl)amino)-1164 benzyl)-1H-pyrazole-5-carboxamide (50). N,4-Dimethylaniline1165 (2.48 mmol) was dissolved in 1,4-dioxane in a microwave tube,1166 followed by boc-protected (4-bromophenyl)methanamine (2.971167 mmol), rac-BINAP (0.25 mmol), and Cs2CO3 (4.95 mmol). The1168 reaction mixture was degassed for 0.5 h before Pd(OAc)2 (0.121169 mmol) was added. The tube was then sealed and placed in a1170 microwave reactor to react at 110 °C for 1 h. Upon completion, the

1171reaction mixture was diluted with EtOAc, washed with NaHCO3 and1172brine, dried over MgSO4, and concentrated in vacuo to afford crude1173product, which was purified by column chromatography (5−20%1174EtOAc/petroleum benzine) to give tert-butyl (4-(methyl(p-tolyl)-1175amino)benzyl)carbamate. This resulting product was then reacted1176with 4 M HCl in 1,4-dioxane at 60 °C for 2 h to give the1177corresponding benzylamine as a free base, which was subsequently1178coupled to 4-chloro-3-ethyl-1-methyl-1H-pyrazole-5-carboxylic acid1179according to General procedure C1 to give the title compound as a1180yellow oil (19%). 1H NMR (400 MHz, CDCl3) δ = 7.12 (d, J = 8.71181Hz, 2H), 7.05 (d, J = 8.1 Hz, 2H), 6.96−6.91 (m, 2H), 6.86 (s, br,11821H), 6.82−6.76 (m, 2H), 4.46 (d, J = 5.5 Hz, 2H), 4.06 (s, 3H), 3.201183(s, 3H), 2.59−2.51 (m, 2H), 2.24 (s, 3H), 1.15 (t, J = 7.6 Hz, 3H)1184ppm; 13C NMR (101 MHz, CDCl3) δ = 158.5, 149.6, 149.0, 146.4,1185132.8, 131.2, 130.1, 128.7, 128.0, 123.3, 117.7, 107.6, 43.2, 40.7, 40.4,118620.8, 19.3, 12.9 ppm; LC-MS: m/z = 396.9 [M + H]+.11874-Chloro-3-ethyl-1-methyl-N-(1-(4-(p-tolyloxy)phenyl)-1188ethyl)-1H-pyrazole-5-carboxamide (51). General procedure C11189was followed, starting from 1-(4-bromophenyl)ethan-1-amine HCl1190and 4-chloro-3-ethyl-1-methyl-1H-pyrazole-5-carboxylic acid to give1191N-(1-(4-bromophenyl)ethyl)-4-chloro-3-ethyl-1-methyl-1H-pyrazole-11925-carboxamide, which was then coupled to p-cresol according to1193General procedure E1 to give the title compound as a yellow oil1194(12%). 1H NMR (400 MHz, CDCl3) δ = 7.30 (d, J = 8.5 Hz, 2H),11957.14 (d, J = 8.2 Hz, 2H), 7.06 (d, J = 7.3 Hz, 1H), 6.94 (dd, J = 19.5,11968.5 Hz, 4H), 5.28−5.19 (m, 1H), 4.13 (s, 3H), 2.67 (q, J = 7.6 Hz,11972H), 2.34 (s, 3H), 1.60 (d, J = 6.9 Hz, 3H), 1.23 (t, J = 7.6 Hz, 3H)1198ppm; 13C NMR (101 MHz, CDCl3) δ = 157.6, 157.6, 154.4, 150.2,1199136.5, 133.3, 131.5, 130.4, 127.5, 119.4, 118.5, 108.3, 49.3, 40.2, 22.3,120020.8, 19.0, 13.0 ppm; LC-MS: m/z = 397.9 [M + H]+.12014-Fluoro-1-methyl-N-(4-(4-methylbenzyl)benzyl)-1H-pyra-1202zole-5-carboxamide (52). General procedure B2 was followed,1203starting from 88 to give the corresponding benzylamine, which was1204subsequently coupled to 4-fluoro-1-methyl-1H-pyrazole-5-carboxylic1205acid according to General procedure C1 to give the title compound as1206a white solid (28%). 1H NMR (400 MHz, CDCl3) δ = 7.33 (d, J = 4.51207Hz, 1H), 7.28−7.24 (m, 2H), 7.19 (d, J = 8.2 Hz, 2H), 7.13−7.06 (m,12084H), 6.51 (d, J = 4.2 Hz, 1H), 4.59 (d, J = 5.7 Hz, 2H), 4.17 (d, J =12091.0 Hz, 3H), 3.94 (s, 2H), 2.32 (s, 3H) ppm; 13C NMR (101 MHz,1210CDCl3) δ = 157.7 (d, J = 4.2 Hz), 150.0, 147.5, 141.2, 137.9, 135.7,1211135.3, 129.4, 129.3, 128.8, 127.9, 124.3 (d, J = 13.4 Hz), 43.0, 41.2,121240.8, 21.1 ppm; LC-MS: m/z = 337.9 [M + H]+.12134-Fluoro-1,3-dimethyl-N-(4-(4-methylbenzyl)benzyl)-1H-1214pyrazole-5-carboxamide (53). General procedure B2 was followed,1215starting from 88 to give the corresponding benzylamine, which was1216subsequently coupled to 4-fluoro-1,3-dimethyl-1H-pyrazole-5-carbox-1217ylic acid according to General procedure C1 to give the title1218compound as a white solid (46%). 1H NMR (400 MHz, CDCl3) δ =12197.25−7.21 (m, 2H), 7.16 (d, J = 8.2 Hz, 2H), 7.11−7.03 (m, 4H),12206.50 (d, J = 5.3 Hz, 1H), 4.56 (d, J = 5.7 Hz, 2H), 4.08 (d, J = 0.8 Hz,12213H), 3.92 (s, 2H), 2.30 (s, 3H), 2.20 (s, 3H) ppm; 13C NMR (1011222MHz, CDCl3) δ = 157.9 (d, J = 4.3 Hz), 148.0, 145.5, 141.1, 137.9,1223135.7, 135.3, 132.7 (d, J = 11.7 Hz), 129.3, 129.2, 128.8, 127.8, 42.9,122441.2, 40.1, 21.0, 9.6 (d, J = 3.1 Hz) ppm; LC-MS: m/z = 351.9 [M +1225H]+.12264-Chloro-1-methyl-N-(4-(4-methylbenzyl)benzyl)-1H-pyra-1227zole-5-carboxamide (54). General procedure B2 was followed,1228starting from 88 to give the corresponding benzylamine, which was1229subsequently coupled to 4-chloro-1-methyl-1H-pyrazole-5-carboxylic1230acid according to General procedure C1 to give the title compound as1231a white solid (57%). 1H NMR (400 MHz, CDCl3) δ = 7.34 (s, 1H),12327.18 (d, J = 8.2 Hz, 2H), 7.10 (d, J = 8.2 Hz, 2H), 7.03−6.96 (m,12334H), 6.88 (s, br, 1H), 4.51 (d, J = 5.7 Hz, 2H), 4.10 (s, 3H), 3.85 (s,12342H), 2.23 (s, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ = 158.3,1235141.2, 137.9, 136.7, 135.7, 135.0, 131.0, 129.4, 129.3, 128.8, 127.9,1236109.6, 43.4, 41.2, 41.2, 21.1 ppm; LC-MS: m/z = 353.9 [M + H]+.12374-Chloro-1,3-dimethyl-N-(4-(4-methylbenzyl)benzyl)-1H-1238pyrazole-5-carboxamide (55). General procedure B2 was followed,1239starting from 88 to give the corresponding benzylamine, which was1240subsequently coupled to 4-chloro-1,3-dimethyl-1H-pyrazole-5-carbox-



N


1241 ylic acid according to General procedure C1 to give the title1242 compound as a white solid (61%). 1H NMR (400 MHz, CDCl3) δ =1243 7.19−7.15 (m, 2H), 7.09 (d, J = 8.2 Hz, 2H), 7.03−6.96 (m, 4H),1244 6.91 (s, br, 1H), 4.50 (d, J = 5.7 Hz, 2H), 4.04 (s, 3H), 3.84 (s, 2H),1245 2.22 (s, 3H), 2.14 (s, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ =1246 158.5, 144.5, 141.1, 137.9, 135.7, 135.1, 131.1, 129.39, 129.2, 128.8,1247 127.8, 108.4, 43.3, 41.2, 40.6, 21.0, 11.1 ppm; LC-MS: m/z = 367.81248 [M + H]+.1249 4-Fluoro-1-methyl-N-(4-((6-methylpyridin-3-yl)methyl)-1250 benzyl)-1H-pyrazole-5-carboxamide (56). Title compound was1251 prepared according to General procedure C1, starting from 94 and 4-1252 fluoro-1-methyl-1H-pyrazole-5-carboxylic acid to give a colorless oil1253 (47%). 1H NMR (400 MHz, CDCl3) δ = 8.44 (d, J = 1.8 Hz, 1H),1254 7.48 (dd, J = 8.0, 2.2 Hz, 1H), 7.30−7.21 (m, 3H), 7.13 (dd, J = 14.5,1255 8.1 Hz, 3H), 6.54 (d, J = 5.3 Hz, 1H), 4.55 (d, J = 5.8 Hz, 2H), 4.111256 (d, J = 0.9 Hz, 3H), 3.93 (s, 2H), 2.57 (s, 3H) ppm; 13C NMR (1011257 MHz, CDCl3) δ = 157.8, 155.0, 146.7, 139.1, 138.9, 136.1, 134.7,1258 129.3, 128.2, 124.5, 120.9, 120.7, 42.9, 40.7 (d, J = 22.0 Hz), 38.2,1259 22.2 ppm; LC-MS: m/z = 338.9 [M + H]+.1260 4-Fluoro-1,3-dimethyl-N-(4-((6-methylpyridin-3-yl)methyl)-1261 benzyl)-1H-pyrazole-5-carboxamide (57). Title compound was1262 prepared according to General procedure C1, starting from 94 and 4-1263 fluoro-1,3-dimethyl-1H-pyrazole-5-carboxylic acid to give a colorless1264 oil (32%). 1H NMR (401 MHz, CDCl3) δ = 8.61 (s, 1H), 8.01 (d, J =1265 7.1 Hz, 1H), 7.54 (t, J = 8.8 Hz, 1H), 7.31 (d, J = 8.0 Hz, 2H), 7.161266 (d, J = 8.0 Hz, 2H), 6.60 (d, J = 5.7 Hz, 1H), 4.59 (d, J = 5.8 Hz, 2H),1267 4.12−4.06 (m, 5H), 2.80 (s, 3H), 2.22 (s, 3H) ppm; 13C NMR (1011268 MHz, CDCl3) δ = 158.0, 152.1, 145.4, 141.1, 138.7, 137.2, 136.4,1269 132.9 (d, J = 12.1 Hz), 129.5, 128.6, 127.2, 121.1, 120.8, 42.8, 40.1,1270 37.9, 19.3, 9.5 (d, J = 3.0 Hz) ppm; LC-MS: m/z = 352.9 [M + H]+.1271 4-Chloro-1-methyl-N-(4-((6-methylpyridin-3-yl)methyl)-1272 benzyl)-1H-pyrazole-5-carboxamide (58). Title compound was1273 prepared according to General procedure C1, starting from 94 and 4-1274 chloro-1-methyl-1H-pyrazole-5-carboxylic acid to give a white solid1275 (35%). 1H NMR (400 MHz, CDCl3) δ = 8.28 (d, J = 14.6 Hz, 1H),1276 7.36−7.33 (m, 1H), 7.31−7.26 (m, 1H), 7.20 (d, J = 7.8 Hz, 2H),1277 7.09 (d, J = 8.2 Hz, 2H), 6.98 (t, J = 7.6 Hz, 1H), 6.93 (s, br, 1H),1278 4.53 (d, J = 5.7 Hz, 2H), 4.10 (s, 3H), 3.85 (s, 2H), 2.44 (s, 3H)1279 ppm; 13C NMR (101 MHz, CDCl3) δ = 158.3, 156.3, 149.2, 139.8,1280 136.9, 136.7, 135.6, 133.2, 130.9, 129.3, 128.0, 123.2, 109.6, 43.3,1281 41.2, 38.3, 23.9 ppm; LC-MS: m/z = 354.8 [M + H]+.1282 4-Chloro-1,3-dimethyl-N-(4-((6-methylpyridin-3-yl)methyl)-1283 benzyl)-1H-pyrazole-5-carboxamide (59). Title compound was1284 prepared according to General procedure C1, starting from 94 and 4-1285 chloro-1,3-dimethyl-1H-pyrazole-5-carboxylic acid to give a white1286 solid (29%). 1H NMR (400 MHz, CDCl3) δ = 8.35 (d, J = 1.6 Hz,1287 1H), 7.35 (dd, J = 7.9, 2.3 Hz, 1H), 7.29−7.23 (m, 2H), 7.14 (d, J =1288 8.2 Hz, 2H), 7.06 (d, J = 8.0 Hz, 1H), 7.02 (s, br, 1H), 4.58 (d, J = 5.71289 Hz, 2H), 4.11 (s, 3H), 3.91 (s, 2H), 2.51 (s, 3H), 2.21 (s, 3H) ppm;1290

13C NMR (101 MHz, CDCl3) δ = 158.5, 156.2, 149.1, 144.5, 139.7,1291 137.0, 135.7, 133.3, 131.0, 129.3, 128.0, 123.2, 108.4, 43.2, 40.6, 38.3,1292 23.9, 11.1 ppm; LC-MS: m/z = 368.9 [M + H]+.1293 3,5-Dimethyl-1H-pyrazole-3,5-dicarboxylate (60). To a1294 stirred solution of 1H-pyrazole-3,5-dicarboxylic acid (3.5 g, 22.431295 mmol) in EtOH (84 mL) was added SOCl2 (14 mL) at 0 °C. The1296 reaction mixture was stirred at room temperature for 18 h. Upon1297 completion, the reaction was concentrated in vacuo to afford the title1298 compound (3.5 g, 85%) as an off-white solid. 1H NMR (400 MHz,1299 CDCl3): δ = 7.34 (s, 1H), 3.95 (s, 6H) ppm; LCMS m/z = 185.0 [M1300 + H] +.1301 3,5-Dimethyl-1-methyl-1H-pyrazole-3,5-dicarboxylate (61).1302 To a stirred solution of 60 (3.5 g, 19.02 mmol) and K2CO3 (3.94 g,1303 28.53 mmol) in acetone (100 mL) at room temperature, dimethyl1304 sulfate (2 mL, 20.92 mmol) was added. The reaction mixture was1305 stirred at 40 °C for 3 h. After completion, the reaction mixture was1306 filtered and the filtrate was concentrated in vacuo to afford the title1307 compound (3.5 g, 93%) as a white solid. 1H NMR (400 MHz,1308 CDCl3): δ = 7.34 (s, 1H), 4.24 (s, 3H), 3.92 (s, 3H), 3.89 (s, 3H)1309 ppm; LCMS m/z = 199 [M + H]+.

13105-(Methoxycarbonyl)-1-methyl-1H-pyrazole-3-carboxylic1311acid (62). To a stirred solution of 61 (4 g, 20.20 mmol) in 1,4-1312dioxane (16 mL) and water (40 mL), concentrated H2SO4 (0.43 mL,13138.081 mmol) was added dropwise. The reaction mixture was refluxed1314for 24 h. Upon completion, the reaction mixture was concentrated in1315vacuo to afford a gummy liquid, which was dissolved in CHCl3 and1316filtered. Filtrate was concentrated to afford the title compound (1.2 g,131732%) as a white solid. 1H NMR (400 MHz, CDCl3): δ = 7.40 (s, 1H),13184.27 (s, 3H), 3.91 (s, 3H) ppm; LCMS m/z = 185.0 [M + H] +.1319Methyl-3-carbamoyl-1-methyl-1H-pyrazole-5-carboxylate1320(63). A mixture of 62 (1.2 g, 4.22 mmol) and SOCl2 (10 mL) was1321stirred at 80 °C for 2 h. The reaction mixture was concentrated,1322diluted with toluene (10 mL), and ammonia gas was passed into the1323reaction mixture at 0 °C for 2 h. After completion, the reaction1324mixture was quenched by the addition of cold water and extracted1325with 10% MeOH in DCM, dried over anhydrous Na2SO4, and1326concentrated to give the title compound (0.92 g, 77%) as an off-white1327solid, which was used in the next step without purification. 1H NMR1328(400 MHz, DMSO-d6): δ = 7.68 (s, 1H), 7.38 (s, 1H), 7.18 (s, 1H),13294.12 (s, 3H), 3.85 (s, 3H) ppm; LC-MS: m/z = 184 [M + H]+

1330Methyl-3-cyano-1-methyl-1H-pyrazole-5-carboxylate (64).1331To a stirred solution of 63 (0.90 g, 4.89 mmol) in DCM (15 mL)1332was added DIPEA (2.3 mL, 13.21 mmol) at 0 °C. A solution of1333trifluroacetic anhydride (0.78 mL, 5.63 mmol) in DCM (5 mL) was1334then added at 0 °C. The reaction mixture was stirred at 0 °C for 2 h1335and then diluted with DCM. The organic layer was washed with1336saturated sodium bicarbonate solution, 5% citric acid solution, and1337brine; dried over Na2SO4; and concentrated in vacuo to afford a1338gummy liquid, which was purified by column chromatography (10%1339EtOAc in hexane) to afford the title compound (0.80 g, 99%) as off-1340white solid. 1H NMR (400 MHz, DMSO-d6): δ = 7.61 (s, 1H), 4.171341(s, 3H), 3.87 (s, 3H) ppm; LC-MS: m/z = 166 [M + H]+.13423-Cyano-1-methyl-1H-pyrazole-5-carboxylic acid (65). Title1343compound was prepared according to General procedure D1, starting1344from 64 to give an off-white solid (37%). 1H NMR (400 MHz,1345DMSO-d6): δ = 14.02 (s, br, 1H), 7.52 (s, 1H), 4.16 (s, 3H) ppm;1346LC-MS: m/z = 149.9 [M − H]+.1347Methyl 4-Fluoro-1-methyl-1H-pyrazole-5-carboxylate (66).1348To a solution of methyl 1-methyl-1H-pyrazole-5-carboxylate (0.5 g)1349in ACN (7 mL) and acetic acid (1.0 mL) was added Selectfluor (1.371350g). The mixture was heated at 100 °C under microwave irradiation for1351120 min. Selectfluor (1.37 g) was added to the mixture and heated at1352100 °C under microwave irradiation for 60 min. The solvent was1353removed in vacuo (water bath was kept at room temperature to avoid1354loss of product under vacuum as the product is very volatile), and the1355residue was partitioned between DCM (15 mL) and water (25 mL).1356The aqueous layer was further extracted with DCM (2 × 10 mL), and1357the combined organic layers were concentrated in vacuo (water bath1358was kept at room temperature to avoid loss of product under vacuum1359as the product is very volatile). The crude product was purified by a1360flash chromatography column on silica gel, eluting with a gradient of13610−15% EtOAc/petroleum benzine to give the title compound as a1362white solid (0.17 g, 31%). 1H NMR (400 MHz, CDCl3) δ = 7.36 (d, J1363= 4.4 Hz, 1H), 4.13 (d, J = 1.0 Hz, 3H), 3.95 (s, 3H). 19F NMR (3761364MHz, CDCl3) δ = −161.32 (s) ppm; LC-MS: Rt 2.89 min, does not1365ionize.13661-Methyl-4-fluoro-1H-pyrazole-5-carboxylic acid (67). Title1367compound was prepared according to General procedure D2, starting1368from 66 to give a white solid (95% yield). 1H NMR (400 MHz,1369DMSO) δ = 7.60 (d, J = 4.3 Hz, 2H), 4.00 (d, J = 1.0 Hz, 7H). 19F1370NMR (376 MHz, DMSO) δ = −162.96 (s) ppm, LC-MS Rt 1.17 min,1371does not ionize.1372(4-(3-Fluoro-4-methylphenoxy)phenyl)methanamine (68).1373General procedure A2 was followed, starting from 3-fluoro-4-1374methylphenol and 4-fluorobenzonitrile to give 4-(3-fluoro-4-1375methylphenoxy)benzonitrile, which was reduced according to General1376procedure B2 to give the title compound as a gummy liquid (80%).1377

1H NMR (400 MHz, CDCl3): δ 7.28−7.25 (m, 2H), 7.11−7.07 (m,13781H), 6.97 (d, J = 8.0 Hz, 2H), 6.69−6.64 (m, 2H), 3.85 (s, 2H), 2.221379(s, 3H) ppm; LC-MS: m/z = 232 [M + H]+.



O


1380 (4-(2-Fluoro-4-methylphenoxy)phenyl)methanamine (69).1381 General procedure A2 was followed, starting from 2-fluoro-4-1382 methylphenol and 4-fluorobenzonitrile to give 4-(2-fluoro-4-1383 methylphenoxy)benzonitrile, which was reduced according to General1384 procedure B2 to give the title compound as a gummy liquid (75%).1385

1H NMR (400 MHz, CDCl3): δ 7.25−7.22 (m, 2H), 6.99−6.90 (m,1386 5H), 3.82 (s, 2H), 2.23 (s, 3H) ppm; LC-MS: m/z = 232 [M + H]+.1387 4-(4-Fluorophenoxy)benzonitrile (70). Title compound was1388 prepared according to General procedure A2, starting from 4-1389 fluorophenol and 4-fluorobenzonitrile to give a gummy liquid (50%).1390

1H NMR (400 MHz, CDCl3): δ 7.59 (d, J = 8.7 Hz, 2H), 7.12−7.071391 (m, H), 7.05−7.01 (m, 2H), 6.96 (d, J = 8.7 Hz, 2H) ppm; LC-MS:1392 m/z = 214 [M + H]+.1393 2-Amino-4-methoxyphenol (71). To a stirred solution of 4-1394 methoxy-2-nitrophenol (7 g, 41.38 mmol) in EtOAc (15 mL) and1395 MeOH (30 mL), Pd/C (10%, 1 g) was added. The reaction mixture1396 was stirred under H2 for 16 h. Upon completion, the reaction mixture1397 was filtered through a pad of celite and washed with EtOAc. The1398 filtrate was concentrated in vacuo to afford the title compound (4.5 g,1399 78%) as a white solid. 1H NMR (400 MHz, DMSO-d6): δ = 8.43 (s,1400 1H), 6.50 (d, J = 8.4 Hz, 1H), 6.20 (d, J = 2.7 Hz, 1H), 5.94 (dd, J =1401 8.3, 2.9 Hz, 1H), 4.52 (s, 2H), 3.57 (s, 3H) ppm; LC-MS: m/z =1402 140.0 [M + H]+.1403 5-Methoxybenzo[d]oxazole (72). A stirred solution of 71 (4.51404 g, 32.40 mmol) in triethyl orthoformate (50 mL) was refluxed for 161405 h. Upon completion, the reaction mixture was cooled to room1406 temperature and concentrated in vacuo. Residue was diluted with1407 EtOAc, and the organic layer was washed with water and brine, dried1408 over anhydrous Na2SO4, and concentrated in vacuo. Crude product1409 was purified by column chromatography (50% EtOAc in hexane) to1410 afford the title compound as a yellow solid (3.6 g, 75%). 1H NMR1411 (400 MHz, CDCl3): δ = 8.04 (s, 1H), 7.44 (d, J = 8.9 Hz, 1H), 7.241412 (d, J = 2.4 Hz, 1H), 6.97 (dd, J = 8.9, 2.5 Hz, 1H), 3.83 (d, J = 6.6 Hz,1413 3H) ppm; LCMS: m/z = 149.9 [M + H]+.1414 Benzo[d]oxazol-5-ol (73). A stirred solution of 72 (3 g, 20.131415 mmol) in DCM (30 mL) was cooled to −20 °C. BBr3 (100 mL,1416 100.70 mmol, 1 M in DCM) was then added dropwise to the reaction1417 mixture. The reaction was stirred at room temperature for 16 h. Upon1418 completion, the reaction mixture was quenched with MeOH and1419 extracted with DCM. The organic layer was washed with water and1420 brine, dried over anhydrous Na2SO4, and concentrated in vacuo.1421 Crude product was purified by column chromatography (50% EtOAc1422 in hexane) to afford the title compound as brown solid (0.75 g, 28%).1423

1H NMR (400 MHz, DMSO-d6): δ = 9.49 (s, 1H), 8.59 (s, 1H), 7.531424 (d, J = 8.7 Hz, 1H), 7.06 (d, J = 1.6 Hz, 1H), 6.86−6.84 (m, 1H)1425 ppm; LC-MS: m/z = 136.0 [M + H]+.1426 4-(Benzo[d]oxazol-5-yloxy)benzonitrile (74). Title compound1427 was prepared according to General procedure A1, starting from 731428 and 4-fluorobenzonitrile to give a white solid (61%). 1H NMR (4001429 MHz, CDCl3): δ = 8.15 (s, 1H), 7.62−7.58 (m, 3H), 7.49 (d, J = 2.31430 Hz, 1H), 7.13 (dd, J = 8.7, 2.3 Hz, 1H), 7.00−6.97 (m, 2H) ppm;1431 LCMS: m/z = 237.1 [M + H]+.1432 Ethyl 4-(4-Cyanophenoxy)benzoate (75). Title compound was1433 prepared according to General procedure G1, starting from ethyl 4-1434 hydroxybenzoate and (4-cyanophenyl)boronic acid to give a brown1435 solid (52%). 1H NMR (400 MHz, DMSO-d6): δ = 8.0. (d, J = 8.6 Hz,1436 2H), 7.90 (d, J = 8.7 Hz, 2H), 7.24 (t, J = 8.3 Hz, 4H), 4.31 (q, J = 7.11437 Hz, 2H), 1.31 (t, J = 7.0 Hz, 3H); ppm; LC-MS: m/z = 268.3 [M +1438 H]+.1439 Ethyl 4-(4-((4-Chloro-3-ethyl-1-methyl-1H-pyrazole-5-1440 carboxamido)methyl)phenoxy)benzoate (76). General proce-1441 dure B3 was followed, starting from 75 to give the desired1442 benzylamine, which was subsequently coupled to 4-chloro-3-ethyl-1-1443 methyl-1H-pyrazole-5-carboxylic acid according to General procedure1444 C3 to afford the title compound as a brown solid (39%). 1H NMR1445 (400 MHz, DMSO-d6): δ = 8.95 (t, J = 6.0 Hz, 1H), 7.95 (d, J = 8.71446 Hz, 2H), 7.42 (d, J = 8.4 Hz, 2H), 7.11 (d, J = 8.4 Hz, 2H), 7.03 (d, J1447 = 8.8 Hz, 2H), 4.50 (d, J = 6.0 Hz, 2H), 4.29 (q, J = 7.0 Hz, 2H), 3.841448 (s, 3H), 2.57−2.50 (m, 2H), 1.30 (t, J = 7.0 Hz, 3H), 1.16 (t, J = 7.61449 Hz, 3H) ppm; LC-MS: m/z = 442.3 [M + H]+.

14505-(p-Tolyloxy)picolinonitrile (77). Title compound was pre-1451pared according to General procedure A2, starting from p-cresol and14525-fluoropicolinonitrile to give a white solid (52%). 1H NMR (4001453MHz, DMSO-d6): δ 8.50 (s, 1H), 8.00 (d, J = 8.6 Hz, 1H), 7.44−7.411454(m, 1H) 7.30 (d, J = 7.7 Hz, 2H), 7.11 (d, J = 8.0 Hz, 2H), 2.33 (s,14553H) ppm; LC-MS: m/z = 210.8 [M + H]+.1456(5-(p-Tolyloxy)pyridin-2-yl)methanamine HCl (78). Title1457compound was prepared according to General procedure B5, starting1458from 77 to give a white solid (34%). 1H NMR (400 MHz, DMSO1459d6): δ 8.38 (s, br, 3H), 7.50−7.47 (m, 2H), 7.24 (d, J = 7.2 Hz, 2H),14606.97 (d, J = 7.7 Hz, 2H), 4.15 (d, J = 4.2 Hz, 2H), 2.31 (s, 3H) ppm;1461LC-MS: m/z = 215.0 [M + H]+.14622-Fluoro-4-(p-tolyloxy)benzonitrile (79). Title compound was1463prepared according to General procedure G2, starting from 2-fluoro-14644-hydroxybenzonitrile and p-tolylboronic acid to give a colorless oil1465(18%). 1H NMR (400 MHz, DMSO-d6): δ 7.50 (t, J = 8.0 Hz, 1H),14667.22 (d, J = 8.1 Hz, 2H), 6.95 (d, J = 8.2 Hz, 2H), 6.78−6.67 (m,14671H), 6.69 (dd, J = 10.6, 2.0 Hz, 1H), 2.37 (s, 3H) ppm; LC-MS: m/z1468= 228.1 [M + H]+.1469(2-Fluoro-4-(p-tolyloxy)phenyl)methanamine HCl (80). Title1470compound was prepared according to General procedure B5, starting1471from 79 to give a pale yellow solid (73%). 1H NMR (400 MHz,1472DMSO-d6): δ 8.25 (s, br, 3H), 7.53 (t, J = 8.6 Hz, 1H), 7.25 (d, J =14738.3 Hz, 2H), 6.97 (d, J = 8.4 Hz, 2H), 6.91−6.88 (m, 1H), 6.84 (dd, J1474= 8.5, 2.3 Hz, 1H), 4.01 (s, 2H), 2.31 (s, 3H) ppm; LC-MS: m/z =1475232.0 [M + H]+.14766-(p-Tolyloxy)nicotinonitrile (81). Title compound was pre-1477pared according to General procedure A2, starting from p-cresol and14786-chloronicotinonitrile to give a pale yellow solid (92%). 1H NMR1479(400 MHz, CDCl3): δ 8.45 (d, J = 1.7 Hz, 1H), 7.89 (dd, J = 8.6, 2.081480Hz, 1H), 7.23 (d, J = 8.2 Hz, 2H), 7.02−6.94 (m, 3H), 2.37 (s, 3H)1481ppm; LC-MS: m/z = 211 [M + H]+.1482(6-(p-Tolyloxy)pyridin-3-yl)methanamine HCl (82). Title1483compound was prepared according to General procedure B5, starting1484from 81 to give a white solid (52%). 1H NMR (400 MHz, DMSO-1485d6): δ 8.28 (br, 3H), 8.20 (s, 1H), 7.95 (d, J = 9.0 Hz, 1H), 7.22 (d, J1486= 7.9 Hz, 2H), 7.05 (d, J = 8.1 Hz, 1H), 6.99 (d, J = 7.8 Hz, 2H),14874.05-3.95 (m, 2H), 2.32 (s, 3H) ppm; LC-MS: m/z = 215 [M + H]+.14883-Fluoro-4-(p-tolyloxy)benzonitrile (83). Title compound was1489prepared according to General procedure A2, starting from 3,4-1490difluorobenzonitrile and p-cresol to give a colorless oil (85%). 1H1491NMR (400 MHz, CDCl3): δ 7.45 (d, J = 10.0 Hz, 1H), 7.33 (d, J =14928.4 Hz, 1H), 7.20 (d, J = 8.0 Hz, 2H), 6.96−6.88 (m, 3H), 2.36 (s,14933H) ppm; LC-MS: m/z = 228.0 [M + H]+.1494(3-Fluoro-4-(p-tolyloxy)phenyl)methanamine (84). Title1495compound was prepared according to General procedure B2, starting1496from 83 to give a gummy liquid (92%). 1H NMR (400 MHz, CDCl3):1497δ 7.15−7.09 (m, 3H), 7.02−6.93 (m, 2H), 6.86 (d, J = 8.2 Hz, 2H),14983.88 (s, br, 2H), 2.31 (s, 3H) ppm; LC-MS: m/z = 232.1 [M + H]+.14994-(4-Chlorophenoxy)-2-fluorobenzonitrile (85). Title com-1500pound was prepared according to General procedure G2, starting1501from 2-fluoro-4-hydroxybenzonitrile and (4-chlorophenyl)boronic1502acid to give a colorless oil (55%). 1H NMR (400 MHz, CDCl3): δ15037.56−7.52 (m, 1H), 7.39 (d, J = 8.5 Hz, 2H), 7.02 (d, J = 8.5 Hz,15042H), 6.79 (d, J = 8.5 Hz, 1H), 6.73 (d, J = 10.1 Hz, 1H) ppm; LC-1505MS: m/z = 248.1 [M + H]+.1506(4-(4-Chlorophenoxy)-2-fluorophenyl)methanamine (86).1507Title compound was prepared according to General procedure B2,1508starting from 85 to give a gummy liquid (92%). 1H NMR (400 MHz,1509CDCl3): δ 7.33−7.23 (m, 3H), 6.97−6.92 (m, 2H), 6.76−6.67 (m,15102H), 3.90 (s, br, 2H) ppm; LC-MS: m/z = 252.1 [M + H]+.15114-(4-Chlorophenoxy)-3-fluorobenzonitrile (87). Title com-1512pound was prepared according to General procedure A2, starting from15133,4-difluorobenzonitrile and 4-chlorophenol to give a colorless oil1514(75%). 1H NMR (400 MHz, CDCl3): δ 7.48 (d, J = 8.4 Hz, 1H),15157.40−7.35 (m, 3H), 7.00−6.96 (m, 3H) ppm; LC-MS: m/z = 248.01516[M + H]+.15174-(4-Methylbenzyl)benzonitrile (88). A solution of 4-1518(chloromethyl)benzonitrile (1 g, 6.60 mmol) and p-tolylboronic1519acid (1.08 g, 7.92 mmol) in DMF (10.0 mL) and water (2.0 mL) was



P


1520 degassed under N2 for 5 min. K2CO3 (1.82 g, 13.2 mmol) and PdCl21521 (0.12 g, 0.66 mmol) were then added to the reaction mixture. The1522 reaction mixture was stirred at 90 °C for 3 h. Upon completion, the1523 reaction mixture was cooled and diluted with EtOAc. The organic1524 layer was washed with water and brine, dried over anhydrous Na2SO4,1525 and concentrated in vacuo to afford crude product, which was purified1526 by column chromatography (5% EtOAc in hexane) to give the title1527 compound as a white solid (88%). 1H NMR (400 MHz, CDCl3): δ =1528 7.56−7.54 (m, 3H), 7.26 (d, J = 9.0 Hz, 1H), 7.11 (d, J = 7.7 Hz,1529 2H), 7.03 (d, J = 7.7 Hz, 2H), 3.98 (s, 2H), 2.32 (s, 3H) ppm; LC-1530 MS: m/z = 208.0 [M + H]+.1531 4-(p-Tolylthio)benzonitrile (89). Title compound was prepared1532 according to General procedure E2, starting from 4-methylbenzene-1533 thiol and 4-iodobenzonitrile to give a white solid (58%). 1H NMR1534 (400 MHz, CDCl3) δ = 7.38−7.30 (m, 4H), 7.18−7.13 (m, 2H),1535 7.04−7.00 (m, 2H), 2.31 (s, 3H) ppm; 13C NMR (101 MHz, CDCl3)1536 δ = 146.6, 140.0, 135.0, 132.3, 130.8, 126.9, 126.8, 118.9, 108.4, 21.41537 ppm; LC-MS: m/z = 225.9 [M + H]+.1538 N-Methoxy-N,6-dimethylnicotinamide (90). Title compound1539 was prepared according to General procedure C6, starting from 6-1540 methylnicotinic acid and N,O-Dimethylhydroxylamine HCl to give a1541 yellow oil (91%). 1H NMR (400 MHz, CDCl3) δ = 8.70 (s, 1H), 7.791542 (dd, J = 8.1, 2.2 Hz, 1H), 7.07 (d, J = 8.1 Hz, 1H), 3.41 (s, 3H), 3.221543 (s, 3H), 2.45 (s, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ = 167.3,1544 160.5, 148.6, 136.4, 126.7, 122.4, 61.0, 33.0, 24.2 ppm; LC-MS: m/z1545 = 180.9 [M + H]+.1546 (4-Bromophenyl)(6-methylpyridin-3-yl)methanone (91).1547 Title compound was prepared according to General procedure H2,1548 starting from 90 and 1,4-dibromobenzene to give a yellow solid1549 (68%). 1H NMR (400 MHz, CDCl3) δ = 8.77 (d, J = 1.0 Hz, 1H),1550 7.92 (dd, J = 8.1, 2.2 Hz, 1H), 7.61−7.53 (m, 4H), 7.23 (d, J = 8.01551 Hz, 1H), 2.58 (s, 3H) ppm; 13C NMR (101 MHz, CDCl3) δ = 193.8,1552 163.0, 150.6, 137.5, 135.8, 131.9, 131.4, 130.1, 128.2, 123.2, 24.81553 ppm; LC-MS: m/z = 275.8 [M + H]+, 277.8 [M + 2]+.1554 5-(4-Bromobenzyl)-2-methylpyridine (92). Compound 911555 (5.79 mmol), hydrazine monohydrate (57.94 mmol), and KOH1556 (23.18 mmol) were dissolved in ethylene glycol (10 mL), followed by1557 stirring at 150 °C for 1 h. Upon completion, the reaction mixture was1558 cooled to room temperature and diluted with H2O. The mixture was1559 extracted with EtOAc (3 × 20 mL), and combined organic layers were1560 washed with brine, dried (MgSO4), and concentrated in vacuo to give1561 crude product, which was purified by column chromatography (5−1562 30% EtOAc/petroleum benzine) to afford the title compound as a1563 yellow oil (55%). 1H NMR (400 MHz, CDCl3) δ = 8.35 (d, J = 2.01564 Hz, 1H), 7.41−7.36 (m, 2H), 7.30 (dd, J = 8.0, 2.3 Hz, 1H), 7.06−1565 6.98 (m, 3H), 3.86 (s, 2H), 2.50 (s, 3H) ppm; 13C NMR (101 MHz,1566 CDCl3) δ = 156.5, 149.2, 139.2, 136.7, 132.7, 131.7, 130.5, 123.2,1567 120.3, 38.0, 24.0 ppm; LC-MS: m/z = 261.8 [M + H]+, 263.8 [M +1568 2]+.1569 4-((6-Methylpyridin-3-yl)methyl)benzonitrile (93). Com-1570 pound 92 (2.29 mmol), K4Fe(CN)6·3H2O (1.14 mmol), XPhos1571 (0.23 mmol), and KOAc (0.30 mmol) were dissolved in a 1:4 mixture1572 of H2O/1,4-dioxane in a sealed tube charged with a magnetic stirring1573 bar. The mixture was degassed for 0.5 h before Pd(dba)3 was added.1574 The reaction was stirred vigorously (≥1000 rpm) at 100 °C for 1 h.1575 Upon completion, the reaction mixture was cooled to room1576 temperature and then extracted with EtOAc and washed with water,1577 NaHCO3, and brine. The organic layer was dried (MgSO4), and1578 solvent was removed in vacuo to afford crude product, which was1579 purified by column chromatography (5−20% EtOAc/petroleum1580 benzine) to yield the title compound as a yellow solid (84%). 1H1581 NMR (400 MHz, CDCl3) δ = 8.29 (d, J = 1.6 Hz, 1H), 7.53−7.461582 (m, 2H), 7.26 (dd, J = 7.9, 2.3 Hz, 1H), 7.22−7.16 (m, 2H), 7.02 (d,1583 J = 7.9 Hz, 1H), 3.92 (s, 2H), 2.45 (s, 3H) ppm; 13C NMR (1011584 MHz, CDCl3) δ = 156.9, 149.3, 145.8, 136.8, 132.5, 131.7, 129.6,1585 123.3, 118.8, 110.5, 38.7, 24.0 ppm; LC-MS: m/z = 209.0 [M + H]+.1586 (4-((6-Methylpyridin-3-yl)methyl)phenyl)methanamine HCl1587 (94). Title compound was prepared according to General procedure1588 B4, starting from 93 to give a yellow solid (86%). 1H NMR (4001589 MHz, DMSO) δ = 8.70 (d, J = 1.8 Hz, 1H), 8.48 (s, br, 3H), 8.31 (d,

1590J = 8.3 Hz, 1H), 7.82 (d, J = 8.2 Hz, 1H), 7.45 (d, J = 8.0 Hz, 2H),15917.36 (d, J = 8.2 Hz, 2H), 4.13 (d, J = 10.8 Hz, 2H), 3.96 (q, J = 5.61592Hz, 2H), 2.70 (s, 3H); 13C NMR (101 MHz, DMSO) δ = 151.6,1593145.4, 140.1, 139.4, 137.9, 132.5, 129.4, 128.9, 127.4, 41.7, 36.4, 18.81594ppm; LC-MS: m/z = 214.0 [M + H]+.1595Interference Compounds. All final compounds have been1596examined for the presence of substructures classified as Pan Assay1597Interference Compounds (PAINS) using a KNIME workflow.32,33

1598■ ASSOCIATED CONTENT1599*S Supporting Information1600The Supporting Information is available free of charge on the1601ACS Publications website at DOI: 10.1021/acs.jmed-1602chem.8b01789.

1603SMILES molecular formula strings (CSV)

1604■ AUTHOR INFORMATION1605Corresponding Authors1606*E-mail: [email protected]. Tel: +61 3 9731 22831607(R.B.S.).1608*E-mail: [email protected]. Tel: +61 3 9903 90441609(J.B.B.).1610ORCID1611Fei Huang: 0000-0002-5290-24901612Jennifer Keiser: 0000-0003-0290-35211613Andreas Hofmann: 0000-0003-4408-54671614Jonathan B. Baell: 0000-0003-2114-82421615Author Contributions1616The manuscript was written through contributions of all1617authors. All authors have given approval to the final version of1618the manuscript.1619Notes1620The authors declare no competing financial interest.

1621■ ACKNOWLEDGMENTS1622Support from the Australian Research Council (ARC),1623Medicines for Malaria Venture (MMV), and Yourgene1624Bioscience is gratefully acknowledged. Fellowship support1625(J.B.) was from the National Health and Medical Research1626Council of Australia (NHMRC) (2012-2016 Senior Research1627Fellowship #1020411, 2017- Principal Research Fellowship1628#1117602). Also acknowledged are Australian Federal Govern-1629ment Education Investment Fund Super Science Initiative and1630the Victorian State Government, Victoria Science Agenda1631Investment Fund for infrastructure support, and the facilities1632and the scientific and technical assistance of the Australian1633Translational Medicinal Chemistry Facility (ATMCF), Mon-1634ash Institute of Pharmaceutical Sciences (MIPS). ATMCF was1635supported by Therapeutic Innovation Australia (TIA). TIA1636was supported by the Australian Government through the1637National Collaborative Research Infrastructure Strategy1638(NCRIS) program. The Centre for Drug Candidate Opti-1639misation, Monash Institute of Pharmaceutical Sciences, is1640acknowledged for their conduct of the ADME screening1641studies. The Peter McCallum Cancer Centre is acknowledged1642for their conduct of the cytotoxicity studies.

1643■ ABBREVIATIONS1644ACN, acetonitrile; DCM, dichloromethane; LHS, left-hand1645side; RHS, right-hand side; TFP, tolfenpyrad



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