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Novel Octahydropyrrolo[3,4-c]pyrroles are Selective Orexin-2 Antagonists: SAR Leading to a Clinical Candidate Michael A Letavic, Pascal Bonaventure, Nicholas I. Carruthers, Christine Dugovic, Tatiana Koudriakova, Brian Lord, Timothy W Lovenberg, Kiev S Ly, Neelakandha S. Mani, Diane Nepomuceno, Daniel J. Pippel, Michele Rizzolio, Jonathan E Shelton, Chandra R Shah, Brock Shireman, Lana K Young, and Sujin Yun J. Med. Chem., Just Accepted Manuscript • Publication Date (Web): 18 Jun 2015 Downloaded from http://pubs.acs.org on June 18, 2015

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Journal of Medicinal Chemistry

Novel Octahydropyrrolo[3,4-c]pyrroles are Selective Orexin-2 Antagonists: SAR Leading to a Clinical Candidate Michael A. Letavic, Pascal Bonaventure, Nicholas I. Carruthers, Christine Dugovic, Tatiana Koudriakova, Brian Lord, Timothy W. Lovenberg, Kiev S. Ly, Neelakandha S. Mani, Diane Nepomuceno, Daniel J. Pippel, Michele Rizzolio, Jonathan E. Shelton, Chandra R. Shah, Brock T. Shireman, Lana K. Young and Sujin Yun Janssen Pharmaceutical Research & Development L.L.C., 3210 Merryfield Row, San Diego, CA 92121

KEYWORDS: Selective Orexin-2 Antagonists, Octahydropyrrolo[3,4-c]pyrroles, Primary Insomnia, JNJ-42847922. ABSTRACT: The pre-clinical characterization of novel octahydropyrrolo[3,4-c]pyrroles that are potent and selective orexin-2 antagonists is described. Optimization of physicochemical and DMPK properties led to the discovery of compounds with tissue distribution and duration of action suitable for evaluation in the treatment of primary insomnia. These selective orexin-2 antagonists are proven to promote sleep in rats and this work ultimately led to the identification of a compound that progressed into human clinical trials for the treatment of primary insomnia.

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The synthesis, SAR and optimization of the pharmacokinetic properties of this series of compounds as well as the identification of the clinical candidate, JNJ-42847922, are described herein. INTRODUCTION Orexin-A and -B are excitatory neuropeptides that originate in orexinergic neurons situated in the hypothalamus, and play important roles in the regulation of sleep/wake cycles, circadian rhythms, energy metabolism, reward-directed behavior, anxious arousal, and stress responses via a relatively discrete network of neuroanatomical projections.1 The orexin (OX) neuropeptides mediate their effect by stimulating two distinct G-protein coupled receptors, orexin-1 (OX1R) and orexin-2 (OX2R) that are co-located or selectively located in specific brain areas suggesting differentiated roles. The salient narcoleptic phenotype of prepro orexin knockout mice2 and the altered orexin signaling in human narcoleptic patients3 provided both genetic and clinical evidence of the key role of this system for the maintenance of wakefulness. This data prompted numerous drug discovery efforts which have recently been disclosed with the goal of understanding how the orexin system functions and whether modulation of this function could produce therapeutic benefit. These research efforts led to the development of several dual OX1/2 receptor antagonists4 including almorexant5 (1) and MK-43056 (2, Suvorexant) (Chart 1) and both of these compounds progressed into human clinical trials and have been shown to promote sleep in humans. Recently, the U.S. Food and Drug Administration approved Belsomra® (2, Suvorexant) for the treatment of primary insomnia.7


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Chart 1. Dual OX1R/OX2R Clinical Candidates.

OBJECTIVES Our efforts in this area have focused on the discovery and development of selective8 orexin-2 antagonists following our observation, using the selective OX2R antagonist JNJ-10397049 (3), that blockade of the OX2R alone is sufficient to initiate and prolong sleep in rodents.9 In additional rat sleep studies, we also demonstrated that pharmacological blockade of the OX1R in the presence of an OX2R antagonist elicited a dysregulation of REM sleep by shifting the balance in favor of REM sleep at the expense of NREM sleep.10 While several OX1R antagonists have been reported to minimally affect spontaneous sleep-wake states,11 controversial data has been found in one study with SB-33486712 however SB-334867 is also known to exhibit off target activities.11 As such, we were most interested in identifying novel, selective OX2R antagonists that were suitable for clinical development and a part of these efforts are now detailed in this report.


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Chart 2. Lead Compounds Used to Identify Selective OX2R Antagonists.

JNJ-10397049 (Chart 2), and similar compounds, provided potential starting points for the optimization of selective OX2R antagonists, however we quickly found that the 2,2-dimethyl1,3-dioxane moiety present in JNJ-10397049 appeared to be essential for activity and, not surprisingly, was also the primary reason for poor oral absorption observed in this series due the acid labile cyclic ketal.13 So, while JNJ-10397049 was a very useful compound to characterize a selective OX2R antagonist in pre-clinical models of sleep, optimization of this chemotype for oral dosing seemed challenging. The high-level goals of the project at this time were to discover novel, selective OX2R antagonists that were orally bioavailable, safe and with an optimal pharmacokinetic profile for the treatment of primary insomnia (quick absorption and short halflife). As such, a high-throughput screening campaign was conducted which, among other chemotypes, identified molecules containing the diazepane core as shown in 4 (Chart 2). Subsequent to our screening campaign this commercially available compound was also reported as a lead by a team at Merck.14 Since that time, several compounds have also been reported in the literature as selective OX2R antagonists, including the aryl amides MK-106415 and MK3697.16 Chart 3. MK-1064 and MK-3697.


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In addition, a number of piperidines and azepanes have been disclosed as selective OX2R antagonists with sleep promoting properties in preclinical studies.17 These include 518 and 6,19 both of which were recently reported by Merck. Compound 5 is orally active in rat sleep models and 6 has been shown to be orally efficacious in mouse sleep models. Chart 4. Compounds 5 and 6.

As part of Janssen’s efforts to follow-up on compound 4 we decided to investigate the OX2R SAR and selectivity of compounds with novel core structures in addition to other diamines including a variety of 3,8-diazabicyclo[4.2.0]octanes, 3,6-diazabicyclo[3.2.0]heptanes,20 octahydropyrrolo[3,4-c]pyrroles.21 This manuscript will focus on optimization of the octahydropyrrolo[3,4-c]pyrroles. RESULTS AND DISCUSSION


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In order to rapidly prepare a variety of analogs, the compounds were initially prepared by one of the general methods shown on Scheme 1. As such, first 2-benzyl-octahydro-pyrrolo[3,4c]pyrrole (7) was converted to hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tert-butyl ester (8) by installation of the Boc group followed by hydrogenation. This sequence was then followed by either addition of the heterocycle, deprotection of the Boc group, and coupling with a benzoic acid or by coupling with a benzoic acid, deprotection of the Boc group and addition of a heterocycle. Scheme 1. Synthesis of Compounds 11-40.

a

Reagents and Conditions: (a) (Boc)2O, DCM, (b) 10% Pd/C, HOAc, 70 psi H2, MeOH, (c)

various benzoic acids, HATU (or similar), diisopropyl ethyl amine, DMF, (d) DCM, TFA, (e)various 2-chloroheterocycles, cesium carbonate, DMF. In cases where the benzoic acid or 2-chloroheterocycle was not commercially available these intermediates were prepared by known methods. All the details of the syntheses of compounds 11-15 and 6 are located in the experimental section. One of the initial compounds prepared in this series was compound 11, which is a rather weak OX2R antagonist (OX2R Ki = 1600 nM).22 Iterative SAR led to some improvements in affinity


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(Table 1) and most importantly, yielded compounds with modest selectivity over OX1R (Compd 15, OX2R Ki = 746 nM, OX1R Ki = 10000 nM). Table 1. Binding Data for the Human OX2 and OX1 Receptor for Compounds 11-15.

a

Compd

R1

R2

OX2R Ki (nM)a

OX1R Ki (nM)a

11

6-MeO-

Cl

2200+870

6850b

12

4-MeO-

Cl

510+120

5300b

13

4-MeO-

Me

680+200

>10000b

14

4-MeO-

-OMe

875+84

>10000b

15

4-MeO-

F

746+388

>10000c

Ki’s are the mean of at least three experiments in triplicate, unless otherwise stated. bn=2 (in

triplicate), cn=3 (in triplicate), Ki + s.d. is reported. This initial promising SAR prompted further modifications of both the amide functionality and the benzothiazole group and produced the compounds shown in Table 2. The 2-phenyl and 2thiophene substituted benzamides 17-20 were also promising leads with slight improvements in OX2R affinity. However selectivity was modest at best but when the 2-thiophene moiety was combined with a quinoxaline (23) or a 4,6-dimethylpyrimidine (24) relatively high affinity OXR2 receptor antagonists were obtained and, at least in the case of 24, good selectivity over OX1R was also observed (~50-fold).


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Table 2. Binding Data for the Human OX2 and OX1 Receptor for Compounds 16-24.

Compd

R3

R4

OX2R Ki (nM)a

OX1R Ki (nM)a

16

Ph-

687+107

1398+387

17

Ph-

294+53

925+84

18

2-thiophene-

137+15

267+25

19

Ph-

421+198

765+178

20

Ph-

240+190

1320+1060

21

Ph-

899+150

>10000b

22

1-pyrazole

768+216

>10000c

23

2-thiophene-

106+95

623+282

24

2-thiophene-

9.8+3.8

471+71

a

Ki’s are the mean of at least three experiments in triplicate, unless otherwise stated., bn=2 (in

triplicate), cn=3 (in triplicate), Ki + s.d. is reported.

Unfortunately, compound 24 suffered from poor in vitro microsomal stability (extraction ratio in rat and human microsomes is >0.92 and >0.95, respectively). Given that the physicochemical


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properties of the compound are not ideal (MW=404.5, cLog P=3.69) this data was not too surprising. Nevertheless, this structure provided a viable lead for the series, and therefore it was further profiled in vitro in prior to in vivo evaluation to benchmark the series. Compound 24 is devoid of hERG affinity (IC50 = >10 µM), is a high affinity rat OX2R ligand (Ki = 9.8 nM) and when tested in a commercial panel of 50 ion channel, receptor and transporter assays (EurofinsCEREP, http://www.eurofins.com/) had no significant binding affinity (10000b

28

27+6

557+94

29

41+19

257+66

30

12+5

1640+620

H O F

27

a

N N

N

N H

Ki’s are the mean of at least three experiments in triplicate.

b

n=3 (in triplicate), Ki + s.d. is

reported. Indeed, compound 30 (MW=389.5, cLog P = 2.44) has slightly better calculated physicochemical properties than 24 and is also marginally superior in the in vitro microsomal stability assay (extraction ratio in rat and human 0.87 and 0.79, respectively. After confirming


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the rat OX2R affinity (Ki = 7 nM), this compound was dosed 30 mg/kg p.o. in a rat and exhibited good receptor occupancy (67% occupancy @ 30 min).23 However, a subsequent rat PK experiment demonstrated that further PK improvements were still needed (F = 8%, CL = 52.1 mL/min/kg, i.v. t1/2 = 0.27 h, p.o. Cmax = 148 ng/mL dosed @10 mg/kg p.o. and 1 mg.kg i.v.). In order to address these issues, additional 2-substituted benzamides were prepared and tested with the goal of making additional improvements to the PK properties of these compounds, for example by blocking or reducing metabolism by introducing fluorine substituents in various positions. These data are shown in Table 4. First, focusing on the 2-substituted triazoles 31-34, the addition of a fluorine atom is tolerated in positions 3 through 6 and all the compounds that were prepared in this set have good OX2R selectivity. Table 4. Binding Data for the Human OX2 and OX1 Receptor for Compounds 31-40. Compd

Structure

OX2R Ki (nM)a

OX1R Ki (nM)a

31

14+5

2280+1500

32

10+3

1960+2180

33

30+15

5780+2860


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34

10+1

800+220

35

30+12

3650+1740

36

120+38

680+3110

37

54+31

3090+1670

38

8+3

346+105

39

46+3

5750+2880

77+19

4630+640

H

40

N O N N

N N

N

N H

Cl

a

Ki’s are the mean of at least three experiments in triplicate, Ki + s.d. is reported.

The 1,2,4-triazole 35 is also a high affinity OX2R ligand. Several other substituted triazolobenzamides were also prepared, including the methyl substituted compounds 36-39, and all are also high affinity OX2R ligands, except for the 6-methyl substituted compound 38, which unexpectedly exhibits modest affinity for OX1R as well.


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Selected compounds were evaluated for microsomal stability, hERG binding, functional human OX2R activity (Kb) and rat OX2R binding affinity in order to determine whether they would be suitable candidates for in vivo characterization. These data are shown in Table 5. The compounds have relatively high extraction ratios, low hERG binding, good functional activity and adequate rat affinity, and it became clear at this point that these structures would require differentiation in vivo. Table 5. Select In Vitro Data for Compounds 24, 30-40. RLM Stabilitya

HLM Stabilityb

hERGc

24

>0.92

>0.95

30

0.79

31

Compd

a

hKb

rKi d

e

R.O.f

(nM)

(nM)

>10 µM

10

7.0

26%g

0.87

>10 µM

3.5

2.0

67%h

0.71

0.83

6.0 µM

2.9+1.6

10+2

49%i

32

0.82

0.88

>10 µM

5.3+1.3

14+3

72%h

33

0.81

0.89

>10 µM

32+5

n.t.

n.t.

34

0.85

0.84

>10 µM

1.6+0.6

10.0+0.1

71%i

35

0.51

0.73

>10 µM

11+6

39

0%i

36

0.81

0.92

n.t.

16

n.t.

n.t.

37

0.86

0.91

>10 µM

16

n.t.

n.t.

38

0.83

0.85

6.9 µM

2.5

12

n.t.

39

0.76

0.87

8.1 µM

7.1

38

n.t.

40

0.91

>0.92

n.t.

16

51

n.t.

Stability in rat liver microsomes. Data reported as extraction ratio. bStability in human liver

microsomes. Data reported as extraction ratio. Extraction ratios are calculated as in vitro CL/Q where Q is species specific hepatic blood flow. chERG IC50 as measured in an [3H]-astemizole competition binding assay in HEK-293 cells expressing the hERG channel. dHuman OX2R Kb,


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data are the result of a single experiment or the average of two experiments in triplicate, except where + s.d. is reported (n=3). eRat OX2R Ki. fData shown are % occupancy at the times and doses specified. gAt 30 min. after a 10 mg/kg p.o. dose. hAt 30 min. after a 30 mg/kg p.o. dose. . iAt 15 min. after a 30 mg/kg p.o. dose. The results of a screening receptor occupancy assay are shown in Table 5. In this assay, compounds are dosed orally at the dose noted and OX2R occupancy is measured in an ex-vivo autoradiography experiment9 with the intention of choosing compounds with high levels of OX2R occupancy for further evaluation. The results of these experiments prompted additional in vivo studies on compounds 30-32 and 34 and they were selected for rat PK experiments. Compound 35 was included, in spite of poor receptor occupancy, because of the low rat liver microsome extraction ratio (0.51). The pharmacokinetic data generated are summarized in Table 6. Table 6. Rat Pharmacokinetic Parameters for Compounds 30-32, 34,and 35. Compd

Cl (mL/min/kg)a

Vss(L/kg)b

t1/2 (h)c

%F

30

52.1

0.7

0.27

8%

31

57.0

0.7

0.20

21%

32

65.5

1.5

0.37

18%

34

40.0

0.7

0.30

24%

35

149.6

3.9

0.38

22%

a

Clearance, bvolume of distribution at steady state, ci.v.half-life.

Compounds 30-32 and 34 had rat PK consistent with the microsomal data collected earlier (Table 5) and all had relatively high clearance, moderate volumes of distribution and short i.v. half-lives. Compound 35 has very high clearance which was not predicted by the rat microsomal


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extraction ratio. The key structural difference is the 1,2,4-triazole present in 35, which might be a potential reason for its significantly different behavior. Compounds 31, 32, and 34 had acceptable bioavailability and based on the screening receptor occupancy data these compounds were progressed for further profiling. Additional in vitro profiling data for compounds 31, 32, and 34 are shown in Table 7. The compounds are all permeable with a low propensity for efflux as measured by a Caco-2 assay. All three compounds have relatively high free fractions as measured by protein binding in rat and human, and the two that were tested exhibit a high free fraction in rat brain tissue. In a Cerep assay (a commercial panel of 50 ion channel, receptor and transporter assays (Eurofins-CEREP, http://www.eurofins.com/ ) compounds 31, 32 and 34 had a 1µM

All > 10µM

CYP Inhibd

1A2 3.1 µM

1A2 9.3 µM

All >10 µM


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Reported as Papp x10-6cm/sec. bPreliminary (Tier 1) data. cRat brain tissue binding. dReported as

an IC50 and all other CYP IC50’s were > 10 µM, CYP 2C9, 2C19, 2C8, 1A2, 2D6 and 3A4 were tested. Compounds in this series had moderate CYP inhibition and of the compounds shown in Table 7, only 31 had significant CYP 1A2 inhibition (3.1 µM). Balancing the overall profiles of compounds 31, 32 and 34 and considering the fact that 31 had some CYP and hERG inhibition, and that both 31 and 32 had slightly higher rat clearance as compared to 34, compound 34 was profiled further in order to predict the human dose required for efficacy, and to predict a duration of action for 34 in humans. As a part of this effort a p.o. concentration-response curve for ex-vivo OX2R occupancy for 34 was generated and the data is shown in Figure 1. Compound 34 dose dependently occupies the OX2R and the EC50 for occupancy in this experiment is 171 ng/mL, which corresponds to an oral ED50 of 3 mg/kg. This EC50 is the concentration we used to help predict the dose required for efficacy in human clinical trials. Our goal was to be able to dose high enough to maintain occupancy above 50% for 4-6 hours because >50% occupancy has been shown to be required to induce sleep in rat9 and our target duration of action for this indication is around 6 hours. In rats implanted with telemetry devices for EEG sleep recording, oral administration of 34 (JNJ42847922) at the onset of the dark phase produced a dose-dependent reduction in NREM latency and an increase in NREM sleep time with no significant effect on REM sleep in these experimental conditions (Figure 2). The lowest effective dose to significantly induce NREM sleep was 3 mg/kg, which corresponds to the ED50 for receptor occupancy. Further details on the


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pre-clinical pharmacology of JNJ-42847922, including additional sleep studies in rodent models, will be disclosed in a separate venue.24 Figure 1. Ex Vivo OX2R Occupancy with Compound 34 in Rat: Concentration-Response After Oral Administrationa

a

Data collected 15 minutes post-dose.


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Figure 2. Dose-Response Effects of 34 (1, 3, 10 and 30 mg/kg, p.o.) on Sleep Parameters in Rat.a

a

Data are expressed in minutes (mean ± SEM of the same 8 animals per dose). Latency to

NREM and REM sleep, and duration of NREM and REM sleep were calculated for 2 hours after compound or vehicle administration at the onset of the dark phase. * P50% OX2R occupancy and using sleep EEG to demonstrate target engagement and functional in vivo activity in a rat. Subsequently, a detailed analysis of the pre-clinical DMPK profile of 34 demonstrated that the compound was a viable pre-clinical candidate. The pre-clinical PK data was used to predict the human PK profile and this analysis suggested that 34 whould have human PK properties needed for a drug to treat primary insomnia (i.e., acceptable bioavailability and a relatively short duration of action) and prompted nomination of this compound as a clinical candidate for the treatment of primary insomnia. Finally, the synthesis of 34 was optimized in order to obtain material for preclinical toleration and developability studies that were used to support nomination. Compound 34 has


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completed GLP toxicology studies and several Phase I clinical trials and is now ready to begin efficacy trials. The results of those studies, including the human PK profile for this compound, will be published in the near future.

EXPERIMENTAL SECTION Chemistry. General Methods. Solvents and reagents were used as supplied by the manufacturer. Unless indicated, all reactions were mechanically stirred and run under an atmosphere of N2. Concentrated refers to concentrated using a rotary evaporator under reduced pressure. Mass spectra (MS) were obtained on an Agilent series 1100 MSD using electrospray ionization (ESI) in positive mode. The calculated (calcd.) mass corresponds to the exact mass. Unless specified otherwise, normal-phase silica gel column chromatography (sgc) was performed on silica gel (SiO2) using prepackaged cartridges and the indicated solvents. Nuclear magnetic resonance (NMR) spectra were obtained on Bruker model DRX spectrometers (300 MHz, 500 MHz or 600 MHz). The following procedures were used to prepare compounds 11-15 and 16-40. Representative Procedure A Step 1 5-Benzyl-hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tert-butyl ester To a solution of 2-benzyl-octahydro-pyrrolo[3,4-c]pyrrole (5.62 g, 27.8 mmol) in DCM (100 mL) was added (Boc)2O (6.16 g, 28.2 mmol). The reaction mixture was stirred for 24 hours at


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23 °C. The solvent was removed in vacuo and the resulting product was used in the next step without further purification. MS (ESI) mass calcd. for C18H26N2O2, 302.41; m/z found, 303.2 [M+H]+. 1H NMR (400 MHz, CDCl3): 7.36 – 7.20 (m, 5H), 3.61 – 3.46 (m, 4H), 3.24 (br s, 2H), 2.85 – 2.72 (m, 2H), 2.70-2.63 (m, 2H), 2.43 – 2.30 (m, 2H), 1.50 – 1.42 (s, 9H). Step 2 Hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tert-butyl ester(8) 5-Benzyl-hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tert-butyl ester (19.85 g, 65.6 mmol), MeOH (200 mL), HOAc (3 mL) and 10% Pd/C Degussa type (400 mg) were charged to a Parr shaker vial and shaken for 3 days at 70 psi hydrogen gas. The resulting material was filtered through Celite® and concentrated. The crude mixture was purified by flash column chromatography (FCC), DCM to 10% MeOH/DCM containing 1% NH4OH, to afford the product. MS (ESI) mass calcd. for C11H20N2O2, 212.29; m/z found, 213.2 [M+H]+. 1H NMR (400 MHz, CDCl3): 3.60-3.55 (m, 2H), 3.38-3.25 (m, 4H), 2.95-2.86 (m, 4H), 1.47 (s, 9H). Step 3 5-Benzothiazol-2-ylhexahydropyrrolo[3,4-c]pyrrole-2-carboxylic acid tert-butyl ester To a solution of hexahydropyrrolo[3,4-c]pyrrole-2-carboxylic acid tert-butyl ester (200 mg, 0.942 mmol) in N,N-dimethylformamide (3 mL) was added 2-chlorobenzothiazole (141 µL, 1.08 mmol) and cesium carbonate (353 mg, 1.08 mmol) and the reaction mixture was stirred at 100 °C for 18 h under argon. The reaction mixture was poured into water (15 mL) and extracted with EtOAc (3 x 8 mL). The combined organic layers were dried over sodium sulfate, filtered and evaporated. The residue was purified by silica gel column chromatography eluting with hexane:


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EtOAc (3:1 to 2:1) to give the title compound (265 mg, 0.767 mmol, 82%) as a white powder. LCMS: 100%, tR = 1.614 min, m/z = 346 [M + H]+. Step 4 2-(Hexahydropyrrolo[3,4-c]pyrrol-2-yl)-benzothiazole A mixture of 5-benzothiazol-2-yl-hexahydropyrrolo[3,4-c]pyrrole-2-carboxylic acid tert-butyl ester (227 mg, 0.657 mmol) and hydrogen chloride (3.18 M in 1,4-dioxane, 2.3 mL, 7.31 mmol) was stirred at room temperature for 1 h. The reaction mixture was diluted with diethyl ether (15 mL) and the precipitate was collected. The product was washed with diethyl ether (3 x 10 mL) to give the title compound (224 mg, 0.704 mmol, >100%) as a white powder. LCMS: 100%, tR = 0.761 min, m/z = 246 [M + H]+. Representative Procedure B 5-Methyl-2-[1,2,3]triazol-2-ylbenzoic acid A suspension of 2-iodo-5-methylbenzoic acid (1.0 g, 3.82 mmol), 1H-1,2,3-triazole (555 µL, 9.58 mmol), copper(I) iodide (65 mg, 0.341 mmol), trans-N,N′-dimethylcyclohexane-1,2diamine (102 µL, 0.647 mmol) and cesium carbonate (2.10 g, 6.44 mmol) in N,Ndimethylformamide (3.5 mL) was stirred at 100 °C for 10 min under microwave irradiation. The reaction mixture was diluted with water (15 mL) and extracted with EtOAc (10 mL). The aqueous layer was acidified to pH 4 by addition of 1 M hydrochloric acid and extracted with EtOAc (6 x 30 mL). The combined organic layers were washed with water (30 mL), dried over sodium sulfate, filtered and evaporated. The residue was purified by silica gel column chromatography eluting with DCM:HOAc (100:1). The product was triturated with hexane (2


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mL) to afford the title compound (440 mg, 2.165 mmol, 57%) as a white crystalline solid. LCMS: 96%, tR = 1.148 min, m/z = 204 [M + H]+, 202 [M − H]−. Representative Procedure C [5-(6-Chlorobenzothiazol-2-yl)-hexahydropyrrolo[3,4-c]pyrrol-2-yl]-(2,6dimethoxyphenyl)-methanone, (11) To a solution of the appropriate amine (100 mg, 0.357 mmol) in N,N-dimethylformamide (2.5 mL) was added 2,6-dimethoxybenzoic acid (72 mg, 0.395 mmol), N,N-diisopropylethylamine (124 µL, 0.712 mmol) and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU, 150 mg, 0.394 mmol) and the reaction mixture was stirred at room temperature for 2 h under argon. The reaction mixture was poured into water (13 mL) and extracted with DCM (3 x 5 mL). The combined organic layers were washed with 5% aqueous sodium carbonate (5 mL), dried over sodium sulfate, filtered and evaporated. The residue was purified by silica gel column chromatography eluting with EtOAc. The product was triturated with EtOAc (1 mL) to afford the title compound (64 mg, 0.144 mmol, 40%) as a white powder. LCMS: 97%, tR = 1.650 min, m/z = 444 [M + H]+. Representative Procedure D (2,4-Dimethoxyphenyl)-[5-(6-fluorobenzothiazol-2-yl)-hexahydropyrrolo[3,4-c]pyrrol-2-yl]methanone, (15) To a mixture of 6-fluoro-2-(hexahydropyrrolo[3,4-c]pyrrol-2-yl)-benzothiazole (200 mg, 0.595 mmol) in water (1.5 mL) and DCM (0.7 mL) was added potassium carbonate (329 mg, 2.38 mmol). To the reaction mixture was then added freshly prepared 2,4-dimethoxybenzoyl chloride


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(0.74 M in DCM, 0.8 mL, 0.592 mmol) dropwise and the mixture was stirred at room temperature for 1 h. The reaction mixture was diluted with DCM (20 mL) and washed with water (5 mL), 1N sodium hydroxide (5 mL) and water (5 mL), dried over sodium sulfate, filtered and evaporated. The residue was purified by silica gel column chromatography eluting with DCM:ethanol (40:1). The residue was triturated with EtOAc (3 mL) to give the title compound (160 mg, 0.374 mmol, 63%) as a white powder. Representative Procedure E [5-(4,6-Dimethylpyrimidin-2-yl)-hexahydropyrrolo[3,4-c]pyrrol-2-yl]-(4-fluoro-2[1,2,3]triazol-2-ylphenyl)-methanone, (32) To a solution of freshly prepared 4-fluoro-2-[1,2,3]triazol-2-ylbenzoyl chloride (0.515 mmol) in DCM (2 mL) was added 2-(4,6-dimethylpyrimidin-2-yl)-octahydropyrrolo[3,4-c]pyrrole (149 mg, 0.512 mmol), N,N-diisopropylethylamine (355 µL, 2.07 mmol) and 4Å molecular sieves, and the reaction mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (10 mL) and washed with water (5 mL), 1 N sodium hydroxide (5 mL) and water (2 x 5 mL), dried over sodium sulfate, filtered and evaporated. The residue was triturated with diethyl ether (1 mL). The product was recrystallized from acetonitrile (0.6 mL) to give the title compound (93 mg, 0.228 mmol, 45%) as a white crystalline solid. The analytical LC/MS method used is as follows: An Agilent 1200 HPLC was used to asses purity. The HPLC column used was a Kinetex, 2.6 µm, C18, 50 x 2.1 mm, maintained at 40 °C. The HPLC Gradient used was as follows: 1.0 mL/min, 95:5:0.1 water:acetonitrile:formic acid to 5:95:0.1 water:acetonitrile:formic acid in 2.0 min, maintaining for 0.5 min, UV channel 220 nm, SI± alternating scans


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The Analytical HPLC method used to assess compound purity is as follows; The instrument used was a Waters Alliance 2695. The HPLC column used was a Kinetex, 5 µm, C18, 150 x 4.6 mm, ambient temperature. The HPLC Gradient used was as follows: 1.0 mL/min, 95:5:0.1 water:acetonitrile:trifluoroacetic acid to 10:90:0.1 water:acetonitrile: trifluoroacetic acid in 10.0 min, maintaining for 1.0 min. Dual channel detection at 220 and 254 nm. General Procedure A Step 1 To a solution of hexahydropyrrolo[3,4-c]pyrrole-2-carboxylic acid tert-butyl ester (200 mg, 0.942 mmol) in N,N-dimethylformamide (3 mL) was added heteroaryl chloride (1.15 equiv) and cesium carbonate (1.15 equiv) and the reaction mixture was stirred at 100 °C for 18 h under argon. The reaction mixture was poured into water (15 mL) and extracted with EtOAc (3 x 10 mL). The combined organic layers were dried over sodium sulfate, filtered and evaporated. The residue was purified by silica gel column chromatography. The following intermediates were prepared by general procedure A. 5-(6-Chloro-benzothiazol-2-yl)-hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tertbutyl ester. Isolated as a white powder (622 mg, 69%); MS m/z 380 [M + H]+. 5-(6-Methoxy-benzothiazol-2-yl)-hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tertbutyl ester. Isolated as an off-white powder (282 mg, 64%); MS m/z 376 [M + H]+. 5-(6-Methyl-benzothiazol-2-yl)-hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tertbutyl ester. Isolated as a white foam (250 mg, 74%); MS m/z 360 [M + H]+.


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5-(6-Fluoro-benzothiazol-2-yl)-hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tertbutyl ester. Isolated as an off-white powder (779 mg, 65%); MS m/z 364 [M + H]+. 5-Quinoxalin-2-yl-hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tert-butyl ester; Isolated as a pale yellow powder (320 mg, 66%); MS m/z 341 [M + H]+. 5-(4-Phenyl-pyrimidin-2-yl)-hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tert-butyl ester. Isolated as a pale yellow powder (212 mg, 49%); MS m/z 367 [M + H]+. 5-(4-Methyl-pyrimidin-2-yl)-hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tert-butyl ester. Isolated as an off-white powder (237 mg, 66%); MS m/z 305 [M + H]+. 5-(4-Methoxy-pyrimidin-2-yl)-hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tert-butyl ester. Isolated as a light brown powder (175 mg, 58%); MS m/z 321 [M + H]+. 5-(4,6-Dimethoxy-pyrimidin-2-yl)-hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tertbutyl ester. Isolated as a white powder (199 mg, 48%); MS m/z 351 [M + H]+. Step 2 A mixture of Boc-protected amine and hydrogen chloride (3.18 M in 1,4-dioxane, 10 equiv) was stirred at room temperature for 1 h. The reaction mixture was diluted with diethyl ether (15 mL) and the precipitate was collected. The product was washed with diethyl ether (3 x 10 mL) to give the title compound. 6-Chloro-2-(hexahydro-pyrrolo[3,4-c]pyrrol-2-yl)-benzothiazole. Isolated as a white powder (205 mg, 70%); MS m/z 280 [M + H]+.


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2-(Hexahydro-pyrrolo[3,4-c]pyrrol-2-yl)-6-methoxy-benzothiazole dihydrochloride salt. Isolated as a white powder (260 mg, 99%); MS m/z 276 [M + H]+. 2-(Hexahydro-pyrrolo[3,4-c]pyrrol-2-yl)-6-methyl-benzothiazole. Isolated as a white powder (146 mg, 83%); MS m/z 260 [M + H]+. 6-Fluoro-2-(hexahydro-pyrrolo[3,4-c]pyrrol-2-yl)-benzothiazole. Isolated as a pale yellow powder (706 mg, 98%); MS m/z 264 [M + H]+. 2-(Hexahydro-pyrrolo[3,4-c]pyrrol-2-yl)-quinoxaline. Isolated as a pale yellow powder (415 mg, 54%); MS m/z 241 [M + H]+. 2-(4-Phenyl-pyrimidin-2-yl)-octahydro-pyrrolo[3,4-c]pyrrole. Isolated as a light brown powder (185 mg, 94%); MS m/z 267 [M + H]+. 2-(4-Methyl-pyrimidin-2-yl)-octahydro-pyrrolo[3,4-c]pyrrole. Isolated as an orange oil (181 mg, 84%); MS m/z 205 [M + H]+. 2-(4-Methoxy-pyrimidin-2-yl)-octahydro-pyrrolo[3,4-c]pyrrole dihydrochloride salt. Isolated as a white powder (286 mg, 93%); MS m/z 221 [M + H]+. 2-(4,6-Dimethoxy-pyrimidin-2-yl)-octahydro-pyrrolo[3,4-c]pyrrole dihydrochloride salt. Isolated as an orange oil (158 mg, 86%); MS m/z 251 [M + H]+. General Procedure B A suspension of the appropriate 2-iodobenzoic acid (1.0 g), 1H-1,2,3-triazole (2.5 equiv), copper(I) iodide (0.09 equiv), trans-N,N′-dimethylcyclohexane-1,2-diamine (0.17 equiv) and cesium carbonate (2.5 equiv) in N,N-dimethylformamide (3.5 mL) was stirred at 100 °C for 10


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min under microwave irradiation. The reaction mixture was diluted with water (15 mL) and extracted with EtOAc (10 mL). The aqueous layer was acidified to pH 4 by addition of 1 M hydrochloric acid and extracted with EtOAc (6 x 30 mL). The combined organic layers were washed with water (30 mL), dried over sodium sulfate, filtered and evaporated. The residue was purified by silica gel column chromatography. 5-Methyl-2-[1,2,3]triazol-2-yl-benzoic acid. Isolated as a white crystalline solid (440 mg, 57%); MS m/z 204 [M + H]+; 1H NMR (500 MHz, Methanol-d4) δ 7.88 (s, 2H), 7.66 (d, J = 2.1 Hz, 1H), 7.60 (d, J = 8.2 Hz, 1H), 7.50 (dd, J = 8.4, 2.0 Hz, 1H), 2.47 (s, 3H). 5-Methoxy-2-[1,2,3]triazol-2-yl-benzoic acid. Isolated as a dark yellow powder (362 mg, 48%); MS m/z 220 [M + H]+; 1H NMR (300 MHz, Methanol-d4) δ 7.99 (s, 1H), 7.88 (s, 2H), 7.62 (d, J = 8.8 Hz, 1H), 7.40 (d, J = 2.9 Hz, 1H), 7.25 (dd, J = 8.8, 2.9 Hz, 1H), 3.93 (s, 3H). 5-Chloro-2-[1,2,3]triazol-2-yl-benzoic acid. Isolated as a pale yellow powder (383 mg, 48%); MS m/z 222 [M - H]-; 1H NMR (300 MHz, Methanol-d4) δ 7.94 (s, 2H), 7.82 (d, J = 2.3 Hz, 1H), 7.79 (d, J = 8.8 Hz, 1H), 7.71 (dd, J = 8.6, 2.4 Hz, 1H). 2-Methyl-6-[1,2,3]triazol-2-yl-benzoic acid. Isolated as a light brown crystalline solid (488 mg, 62%); MS m/z 204 [M + H]+; 1H NMR (300 MHz, Methanol-d4) δ 7.91 (s, 2H), 7.75 (d, J = 8.0 Hz, 1H), 7.50 (t, J = 7.9 Hz, 1H), 7.38 (d, J = 7.6 Hz, 1H), 2.48 (s, 3H). General Procedure C To a solution of amine (100 mg) in N,N-dimethylformamide (2.5 mL) was added the appropriate benzoic acid (1.1 equiv), N,N-diisopropylethylamine (2 equiv) and 1[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide


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hexafluorophosphate (HATU, 1.1 equiv) and the reaction mixture was stirred at room temperature under argon. The reaction mixture was poured into water (15 mL) and extracted with DCM (3 x 10 mL). The combined organic layers were washed with 5% aqueous sodium carbonate (10 mL), dried over sodium sulfate, filtered and evaporated. The residue was purified by silica gel column chromatography. The compounds were prepared using General Procedure C are as follows. [5-(6-Chloro-benzothiazol-2-yl)-hexahydro-pyrrolo[3,4-c]pyrrol-2-yl]-(2,6-dimethoxyphenyl)-methanone (11). Triturated with EtOAc (1 mL) to give a white crystalline solid (64 mg, 40%); mp 203-204.5 °C; 1H NMR (500 MHz, DMSO-d6) δ 7.90 (d, J = 2.3 Hz, 1H), 7.44 (d, J = 8.6 Hz, 1H), 7.29 (t, J = 8.7 Hz, 1H), 7.28 (d, J = 8.3 Hz, 1H), 6.70 (d, J = 8.4 Hz, 1H), 6.66 (d, J = 8.3 Hz, 1H), 3.77 (dd, J = 10.0, 7.8 Hz, 1H), 3.75 (s, 3H), 3.70 (dd, J = 10.6, 7.5 Hz, 1H), 3.67 – 3.61 (m, 1H), 3.64 (s, 3H), 3.52 (dd, J = 12.5, 4.6 Hz, 1H), 3.43 (dd, J = 10.6, 4.6 Hz, 1H), 3.36 (dd, J = 11.0, 7.3 Hz, 1H), 3.34 – 3.30 (m, 1H), 3.15 – 3.08 (m, 1H), 3.07 – 3.00 (m, 1H), 2.97 (dd, J = 10.8, 4.6 Hz, 1H); 13C NMR (126 MHz, DMSO-d6) δ 164.86, 164.00, 155.99, 155.53, 151.82, 132.06, 130.22, 126.05, 124.37, 120.88, 119.11, 115.49, 104.35, 104.16, 55.70, 55.54, 53.48, 52.94, 50.45, 48.76, 41.60, 40.78; HRMS calcd for C22H23ClN3O3S [M + H]+ 444.1149; Found 444.1152. Anal. Calcd for C22H22ClN3O3S: C, 59.52; H, 4.99; N, 9.47; Found: C, 59.15; H, 4.83; N, 9.18. 6-Chloro-2-{5-[(2,4-dimethoxyphenyl)carbonyl]hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl}1,3-benzothiazole (12). Triturated with EtOAc (1 mL) to give an off-white powder (96 mg, 60%); mp 155-157.5 °C; 1H NMR (500 MHz, DMSO-d6) δ 7.90 (d, J = 2.3 Hz, 1H), 7.43 (d, J = 8.6 Hz, 1H), 7.28 (dd, J = 8.6, 2.2 Hz, 1H), 7.12 (d, J = 8.3 Hz, 1H), 6.59 (d, J = 2.3 Hz, 1H),


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6.53 (dd, J = 8.4, 2.2 Hz, 1H), 3.81 – 3.75 (m, 1H), 3.78 (s, 3H), 3.73 (s, 3H), 3.72 – 3.65 (m, 2H), 3.51 – 3.40 (m, 3H), 3.36 – 3.27 (m, 1H), 3.17 – 3.07 (m, 2H), 3.07 – 2.98 (m, 1H); 13C NMR (126 MHz, DMSO-d6) δ 166.55, 164.87, 161.10, 156.12, 151.80, 132.08, 128.58, 126.04, 124.36, 120.86, 119.53, 119.09, 105.20, 98.40, 55.41, 55.28, 53.51, 52.78, 50.77, 49.21, 41.79, 40.59; HRMS calcd for C22H23ClN3O3S [M + H]+ 444.1149; Found 444.1135. Anal. Calcd for C22H22ClN3O3S: C, 59.52; H, 4.99; N, 9.47; Found: C, 59.36; H, 4.92; N, 9.25. 2-{5-[(2,4-Dimethoxyphenyl)carbonyl]hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl}-6-methyl1,3-benzothiazole (13). Triturated with diisopropyl ether (1 mL) to give an off-white foam (52 mg, 28%); 1H NMR (500 MHz, DMSO-d6) δ 7.56 (s, 1H), 7.35 (d, J = 8.2 Hz, 1H), 7.12 (d, J = 8.3 Hz, 1H), 7.08 (d, J = 8.3 Hz, 1H), 6.59 (s, 1H), 6.54 (d, J = 8.5 Hz, 1H), 3.85 – 3.61 (m, 3H), 3.78 (s, 3H), 3.73 (s, 3H), 3.53 – 3.38 (m, 3H), 3.33 – 3.29 (m, 1H), 3.18 – 3.05 (m, 2H), 3.05 – 2.95 (m, 1H), 2.33 (s, 3H); 13C NMR (126 MHz, DMSO-d6) δ 166.55, 163.70, 161.09, 156.12, 150.80, 130.56, 129.77, 128.58, 126.94, 121.02, 119.55, 117.91, 105.20, 98.40, 55.41, 55.28, 53.47, 52.75, 50.86, 49.29, 41.78, 40.58, 20.68; HRMS calcd for C23H26N3O3S [M + H]+ 424.1695; Found 424.1690. 6-Fluoro-2-{5-[(2-thiophen-2-ylphenyl)carbonyl]hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl}1,3-benzothiazole (18). Isolated as an off-white foam (154 mg, 76%); 1H NMR (500 MHz, DMSO-d6) δ 7.70 (dd, J = 8.7, 2.8 Hz, 1H), 7.62 – 7.38 (m, 5H), 7.35 (dd, J = 7.5, 1.4 Hz, 1H), 7.23 – 7.15 (m, 1H), 7.11 (td, J = 9.1, 2.8 Hz, 1H), 7.09 – 6.97 (m, 1H), 3.83 – 3.60 (m, 2H), 3.60 – 2.59 (m, 8H); 13C NMR (126 MHz, DMSO-d6) δ 168.05, 164.12, 156.89 (d, J = 236.5 Hz), 149.55, 140.55, 135.95, 131.41 (d, J = 11.1 Hz), 130.00, 129.35, 128.97, 127.97, 127.95, 127.00, 126.91, 125.92, 118.66 (d, J = 8.8 Hz), 113.28 (d, J = 23.6 Hz), 108.02 (d, J = 27.3 Hz),


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53.12, 52.46, 50.99, 49.21, 41.74, 40.41; HRMS calcd for C24H21FN3OS2 [M + H]+ 450.1110; Found 450.1112. 2-{5-[(2-Thiophen-2-ylphenyl)carbonyl]hexahydropyrrolo[3,4-c]pyrrol-2(1H)yl}quinoxaline (23). Triturated with diisopropyl ether (5 mL) to give a yellow foam (107 mg, 60%); 1H NMR (500 MHz, DMSO-d6) δ 8.35 (s, 1H), 7.82 (d, J = 8.1 Hz, 1H), 7.67 – 7.50 (m, 3H), 7.50 – 7.25 (m, 5H), 7.24 – 7.10 (m, 1H), 7.10 – 6.84 (m, 1H), 3.93 – 3.54 (m, 3H), 3.54 – 3.36 (m, 2H), 3.36 – 3.12 (m, 2H), 3.11 – 2.59 (m, 3H); 13C NMR (126 MHz, DMSO-d6) δ 168.07, 150.15, 141.63, 140.60, 136.92, 135.98, 129.94, 129.81, 129.31, 128.94, 128.45, 127.96, 127.89, 126.89, 125.90, 125.70, 123.52, 51.14, 50.35, 49.66, 49.39, 41.34, 40.08; HRMS calcd for C25H23N4OS [M + H]+ 427.1593; Found 427.1588. 2-(4,6-Dimethylpyrimidin-2-yl)-5-[(2-thiophen-2-ylphenyl)carbonyl]octahydro-pyrrolo[3,4c]pyrrole (24). Purified by preparative HPLC (water:methanol gradient, 1.5% trifluoroacetic acid) to give a pale yellow glass (48 mg, 34%); 1H NMR (500 MHz, DMSO-d6) δ 7.62 – 7.36 (m, 1H), 7.56 (d, J = 7.8 Hz, 1H), 7.47 (td, J = 7.6, 1.6 Hz, 1H), 7.41 (t, J = 7.5 Hz, 1H), 7.34 (d, J = 7.5 Hz, 1H), 7.26 – 6.90 (m, 2H), 6.38 (s, 1H), 3.82 – 2.58 (m, 10H), 2.21 (s, 6H); 13C NMR (126 MHz, DMSO-d6) δ 168.04 , 166.36 , 160.02 , 140.55 , 136.00 , 130.01 , 129.29 , 128.92 , 127.93 , 127.89 , 126.91 , 126.86 , 125.82 , 108.24 , 51.28 , 50.33 , 49.65 , 49.49 , 41.34 , 40.09 , 23.63; HRMS calcd for C23H25N4OS [M + H]+ 405.1749; Found 405.1738. 2-(4,6-Dimethoxypyrimidin-2-yl)-5-[(2-thiophen-2-ylphenyl)carbonyl]octahydro pyrrolo[3,4-c]pyrrole (29). Isolated as a pale yellow foam (125 mg, 58%); 1H NMR (500 MHz, DMSO-d6) δ 7.63 – 7.37 (m, 1H), 7.56 (d, J = 7.8 Hz, 1H), 7.47 (t, J = 7.5 Hz, 1H), 7.41 (t, J = 7.5 Hz, 1H), 7.35 (d, J = 7.5 Hz, 1H), 7.25 – 7.13 (m, 1H), 7.13 – 6.93 (m, 1H), 5.38 (s, 1H),


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3.79 (s, 6H), 3.77 – 2.59 (m, 10H); 13C NMR (126 MHz, DMSO-d6) δ 171.61, 168.56, 159.55, 141.06, 136.49, 130.47, 129.80, 129.43, 128.45, 128.34, 127.40, 127.32, 126.36, 77.86, 53.46, 51.60, 50.56, 49.88, 41.73, 40.40; HRMS calcd for C23H25N4O3S [M + H]+ 437.1647; Found 437.1649. 2-(4,6-Dimethylpyrimidin-2-yl)-5-{[2-(2H-1,2,3-triazol-2-yl)phenyl]carbonyl} octahydropyrrolo[3,4-c]pyrrole (30). Triturated with diisopropyl ether (1 mL) to give a white crystalline solid (71 mg, 53%); mp 159.5-161.8 °C; 1H NMR (500 MHz, DMSO-d6) δ 7.97 (s, 2H), 7.87 (d, J = 8.0 Hz, 1H), 7.61 (td, J = 7.7, 1.8 Hz, 1H), 7.52 (t, J = 7.4 Hz, 1H), 7.49 (dd, J = 7.6, 1.7 Hz, 1H), 6.39 (s, 1H), 3.73 (dd, J = 11.5, 7.6 Hz, 1H), 3.67 (dd, J = 12.4, 7.5 Hz, 1H), 3.57 (dd, J = 11.7, 7.0 Hz, 1H), 3.48 – 3.34 (m, 4H), 3.03 – 2.84 (m, 3H), 2.23 (s, 6H); 13C NMR (126 MHz, DMSO-d6) δ 166.43, 166.39, 160.04, 136.37, 135.31, 130.71, 129.92, 128.48, 127.99, 122.33, 108.20, 51.54, 50.35, 49.79, 49.45, 41.45, 40.16, 23.62; HRMS calcd for C21H24N7O [M + H]+ 390.2042; Found 390.2039. Anal. Calcd for C21H23N7O: C, 64.76; H, 5.95; N, 25.18; Found: C, 64.63; H, 5.58; N, 25.23. 2-(4,6-Dimethylpyrimidin-2-yl)-5-{[3-fluoro-2-(2H-1,2,3-triazol-2-yl)phenyl]carbonyl} octahydropyrrolo[3,4-c]pyrrole (31). Triturated with diethyl ether (1.5 mL) to give a white crystalline solid (125 mg, 59%); mp 151.2-153.2 °C; 1H NMR (500 MHz, DMSO-d6) δ 7.99 (s, 2H), 7.67 (ddd, J = 13.1, 8.2, 5.1 Hz, 1H), 7.62 – 7.55 (m, 1H), 7.41 (d, J = 7.6 Hz, 1H), 6.39 (s, 1H), 3.70 (dd, J = 11.5, 7.2 Hz, 1H), 3.60 (dd, J = 11.7, 6.8 Hz, 1H), 3.54 (dd, J = 12.6, 7.3 Hz, 1H), 3.50 (dd, J = 10.7, 7.3 Hz, 1H), 3.42 (dd, J = 11.7, 3.8 Hz, 1H), 3.35 (dd, J = 11.6, 4.8 Hz, 1H), 3.26 (dd, J = 12.5, 3.9 Hz, 1H), 3.10 (dd, J = 10.8, 4.8 Hz, 1H), 2.99 – 2.88 (m, 2H), 2.23 (s, 6H); 13C NMR (126 MHz, DMSO-d6) δ 166.43, 164.28, 160.10, 155.39 (d, J = 253.9 Hz), 136.43, 136.27, 131.64 (d, J = 8.3 Hz), 124.51 (d, J = 12.1 Hz), 123.25 (d, J = 3.8 Hz), 117.49


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(d, J = 19.4 Hz), 108.23, 51.75, 50.23, 49.70, 49.38, 41.48, 40.10, 23.65; HRMS calcd for C21H23N7OF [M + H]+ 408.1948; Found 408.1944. Anal. Calcd for C21H22N7OF: C, 61.90; H, 5.44; N, 24.06; Found: C, 61.48; H, 5.27; N, 23.63. 2-(4,6-Dimethylpyrimidin-2-yl)-5-{[2-(1H-1,2,4-triazol-5-yl)phenyl]carbonyl} octahydropyrrolo[3,4-c]pyrrole (35). Triturated with diethyl ether (1.5 mL) to give a white powder (52 mg, 26%); Mixture of tautomers by NMR (ca. 4:1). Major tautomer is tabulated: 1H NMR (500 MHz, DMSO-d6) δ 14.09 (s, 1H), 8.50 (s, 1H), 8.07 – 7.21 (m, 4H), 6.37 (s, 1H), 3.84 – 3.64 (m, 2H), 3.64 – 3.18 (m, 5H), 3.06 – 2.74 (m, 3H), 2.22 (s, 6H); All peaks reported: 13C NMR (126 MHz, DMSO-d6) δ 166.41, 160.08, 159.94, 136.61, 128.94, 128.59, 127.91, 127.38, 126.75, 108.21, 51.62, 51.40, 50.34, 49.74, 49.38, 41.47, 40.17, 23.64; HRMS calcd for C21H24N7O [M + H]+ 390.2042; Found 390.2043. General Procedure D To a mixture of amine (150 mg) in DCM (1.4 mL) and water (1.0 mL) was added potassium carbonate (4 equiv). To the reaction mixture was added freshly prepared 2-biphenylcarbonyl chloride (solution in DCM, 1 equiv) dropwise and the mixture was stirred at room temperature for 1 h. The reaction mixture was diluted with DCM (15 mL) and washed with water (5 mL), 1N sodium hydroxide (5 mL) and water (5 mL), dried over sodium sulfate, filtered and evaporated. The residue was purified by silica gel column chromatography. The compounds that were prepared using General Procedure D are as follows. 2-{5-[(2,4-Dimethoxyphenyl)carbonyl]hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl}-6-methoxy1,3-benzothiazole (14). Triturated with EtOAc (1.5 mL) to give a white powder (111 mg, 34%); mp 107.5-111.5 °C; 1H NMR (500 MHz, DMSO-d6) δ 7.40 (s, 1H), 7.37 (d, J = 8.9 Hz, 1H),


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7.12 (d, J = 8.3 Hz, 1H), 6.88 (d, J = 8.7 Hz, 1H), 6.59 (s, 1H), 6.54 (d, J = 8.4 Hz, 1H), 3.83 – 3.59 (m, 3H), 3.78 (s, 3H), 3.75 (s, 3H), 3.73 (s, 3H), 3.52 – 3.37 (m, 3H), 3.34 – 3.25 (m, 1H), 3.17 – 2.94 (m, 3H); 13C NMR (126 MHz, DMSO-d6) δ 166.53, 162.91, 161.09, 156.12, 154.08, 147.04, 131.46, 128.57, 119.55, 118.60, 113.54, 105.57, 105.19, 98.40, 55.51, 55.41, 55.27, 53.46, 52.73, 50.88, 49.30, 41.78, 40.59; HRMS calcd for C23H26N3O4S [M + H]+ 440.1644; Found 440.1644. Anal. Calcd for C23H25N3O4S: C, 62.85; H, 5.73; N, 9.56; Found: C, 62.64; H, 5.48; N, 9.43. 6-Fluoro-2-{5-[(2,4-dimethoxyphenyl)carbonyl]hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl}1,3-benzothiazole (15). Triturated with EtOAc (3 mL) to give a white crystalline solid (160 mg, 63%); mp 169-170 °C; 1H NMR (500 MHz, DMSO-d6) δ 7.70 (dd, J = 8.7, 2.7 Hz, 1H), 7.44 (dd, J = 8.9, 4.8 Hz, 1H), 7.12 (d, J = 8.2 Hz, 1H), 7.11 (dd, J = 9.0, 2.8 Hz, 1H), 6.59 (d, J = 2.4 Hz, 1H), 6.54 (dd, J = 8.3, 2.2 Hz, 1H), 3.81 – 3.64 (m, 4H), 3.78 (s, 3H), 3.73 (s, 3H), 3.51 – 3.40 (m, 3H), 3.15 – 3.06 (m, 2H), 3.06 – 2.98 (m, 1H); 13C NMR (126 MHz, DMSO-d6) δ 166.56, 164.28, 161.10, 156.90 (d, J = 236.6 Hz), 156.13, 149.60, 131.38 (d, J = 10.9 Hz), 128.58, 119.54, 118.68 (d, J = 8.9 Hz), 113.32 (d, J = 23.8 Hz), 108.05 (d, J = 27.3 Hz), 105.21, 98.40, 55.42, 55.28, 53.48, 52.75, 50.82, 49.25, 41.80, 40.60; HRMS calcd for C22H23FN3O3S [M + H]+ 428.1444; Found 428.1440. Anal. Calcd for C22H22FN3O3S: C, 61.81; H, 5.19; N, 9.83; Found: C, 61.74; H, 4.91; N, 9.51. Biphenyl-2-yl-[5-(6-methyl-benzothiazol-2-yl)-hexahydro-pyrrolo[3,4-c]pyrrol-2-yl]methanone (16). Isolated as an off-white foam (86 mg, 43%); 1H NMR (500 MHz, DMSO-d6) δ 7.77 (d, J = 7.7 Hz, 1H), 7.55 – 7.19 (m, 11H), 7.05 (td, J = 7.5, 1.2 Hz, 1H), 3.70 – 3.50 (m, 2H), 3.50 – 3.35 (m, 1H), 3.31 – 2.53 (m, 7H); 13C NMR (126 MHz, DMSO-d6) δ 168.51, 164.08, 152.88, 139.46, 137.61, 136.55, 130.56, 129.38, 129.36, 128.32, 128.06, 127.81, 127.64,


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126.94, 125.84, 121.13, 120.61, 118.22, 52.97, 52.36, 50.72, 49.05, 41.64, 40.25; HRMS calcd for C26H24N3OS [M + H]+ 426.1640; Found 426.1634. 2-[5-(Biphenyl-2-ylcarbonyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl]-6-fluoro-1,3benzothiazole (17). Isolated as an off-white foam (142 mg, 72%); 1H NMR (500 MHz, DMSOd6) δ 7.77 – 7.65 (m, 1H), 7.59 – 7.18 (m, 10H), 7.18 – 7.05 (m, 1H), 3.68 – 3.49 (m, 2H), 3.48 – 3.35 (m, 1H), 3.31 – 2.54 (m, 7H); 13C NMR (126 MHz, DMSO-d6) δ 168.50, 164.01, 156.90 (d, J = 236.7 Hz), 149.57, 139.45, 137.59, 136.53, 131.42 (d, J = 11.0 Hz), 129.37, 129.36, 128.30, 128.05, 127.79, 127.63, 126.93, 118.65 (d, J = 8.8 Hz), 113.28 (d, J = 23.8 Hz), 108.02 (d, J = 27.4 Hz), 52.94, 52.34, 50.70, 49.03, 41.63, 40.25; HRMS calcd for C26H23FN3OS [M + H]+ 444.1546; Found 444.1546. 2-(Biphenyl-2-ylcarbonyl)-5-(4-phenylpyrimidin-2-yl)octahydropyrrolo[3,4-c]pyrrole (19). Triturated with EtOAc (1 mL) to give a white powder (114 mg, 47%); mp 120.3-121.1 °C; 1H NMR (500 MHz, DMSO-d6) δ 8.41 (d, J = 5.1 Hz, 1H), 8.21 – 8.08 (m, 2H), 7.59 – 6.95 (m, 12H), 7.20 (d, J = 5.1 Hz, 1H), 3.83 – 3.40 (m, 3H), 3.40 – 2.54 (m, 7H); 13C NMR (126 MHz, DMSO-d6) δ 168.50, 162.87, 160.03, 158.60, 139.46, 137.61, 136.94, 136.65, 130.59, 129.32, 128.66, 128.20, 127.99, 127.59, 126.92, 126.70, 104.96, 50.99, 50.16, 49.53, 49.32, 41.28, 39.92; HRMS calcd for C29H27N4O [M + H]+ 447.2185; Found 447.2169. 2-(Biphenyl-2-ylcarbonyl)-5-(4-methylpyrimidin-2-yl)octahydropyrrolo[3,4-c]pyrrole (21). Tan low-melting solid (125 mg, 50%); 1H NMR (500 MHz, DMSO-d6) δ 8.17 (d, J = 5.0 Hz, 1H), 7.50 (t, J = 7.5 Hz, 1H), 7.46 – 7.40 (m, 2H), 7.40 – 7.16 (m, 6H), 6.49 (d, J = 5.0 Hz, 1H), 3.63 – 3.48 (m, 2H), 3.48 – 3.35 (m, 1H), 3.27 – 3.03 (m, 2H), 3.03 – 2.54 (m, 5H), 2.27 (s, 3H); 13

C NMR (126 MHz, DMSO-d6) δ 168.45, 166.72, 159.83, 157.17, 139.43, 137.65, 136.62,


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129.32, 129.30, 128.23, 127.95, 127.61, 127.55, 126.90, 108.89, 51.03, 50.15, 49.53, 49.29, 41.24, 39.85, 23.83; HRMS calcd for C24H25N4O [M + H]+ 385.2028; Found 385.2030. 2-(4,6-Dimethylpyrimidin-2-yl)-5-[(2-methylnaphthalen-1-yl)carbonyl]octahydropyrrolo[3,4-c]pyrrole (25). Triturated with diisopropyl ether (1 mL) to give a white powder (87 mg, 43%); mp 148.2-149.4 °C; Mixture of atropisomers by NMR (ca. 7:3). Major isomer is tabulated below. 1H NMR (500 MHz, DMSO-d6) δ 7.95 – 7.88 (m, 1H), 7.86 (d, J = 8.4 Hz, 1H), 7.52 – 7.33 (m, 4H), 6.39 (s, 1H), 3.88 (dd, J = 12.5, 7.8 Hz, 1H), 3.82 – 3.73 (m, 1H), 3.69 (dd, J = 12.5, 4.6 Hz, 1H), 3.58 – 3.50 (m, 2H), 3.35 – 3.26 (m, 1H), 3.22 (dd, J = 11.7, 4.6 Hz, 1H), 3.11 – 3.02 (m, 1H), 2.96 – 2.87 (m, 1H), 2.67 (dd, J = 11.2, 5.0 Hz, 1H), 2.37 (s, 3H), 2.21 (s, 6H); All peaks reported. 13C NMR (126 MHz, DMSO-d6) δ 167.62, 167.56, 166.43, 160.10, 160.08, 134.07, 134.02, 131.24, 130.67, 130.47, 128.59, 128.52, 128.39, 128.02, 126.96, 126.81, 125.42, 125.36, 123.93, 123.71, 108.31, 50.93, 50.71, 50.63, 50.08, 49.98, 49.27, 49.20, 41.38, 40.20, 23.60, 18.78, 18.73; HRMS calcd for C24H27N4O [M + H]+ 387.2185; Found 387.2193. 2-(4-Methoxypyrimidin-2-yl)-5-[(2-methylnaphthalen-1-yl)carbonyl]octahydro-pyrrolo[3,4c]pyrrole (26). Isolated as a pale yellow glass (162 mg, 43%); Mixture of atropisomers by NMR (ca. 1:1). All peaks reported. 1H NMR (500 MHz, DMSO-d6) δ 8.05 (d, J = 5.7 Hz, 1H), 7.97 – 7.88 (m, 1H), 7.86 (d, J = 8.5 Hz, 1H), 7.67 – 7.33 (m, 4H), 6.08 – 6.03 (m, 1H), 4.01 – 3.74 (m, 5H), 3.74 – 3.44 (m, 3H), 3.36 – 3.03 (m, 3H), 2.98 – 2.64 (m, 2H), 2.40 – 2.26 (m, 3H); 13C NMR (126 MHz, DMSO-d6) δ 169.18, 167.67, 167.61, 160.02, 160.00, 158.22, 134.07, 134.00, 131.26, 130.73, 130.45, 128.61, 128.53, 128.42, 128.05, 126.99, 126.82, 125.45, 125.40, 123.95, 123.69, 95.60, 52.59, 50.94, 50.87, 50.62, 50.55, 50.01, 49.88, 49.25, 49.17, 41.36, 40.20, 18.80, 18.74; HRMS calcd for C23H25N4O2 [M + H]+ 389.1978; Found 389.1975.


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2-(4,6-Dimethylpyrimidin-2-yl)-5-[(3'-fluorobiphenyl-2-yl)carbonyl]octahydro-pyrrolo[3,4c]pyrrole (27). Isolated as a pale yellow low-melting solid (168 mg, 78%); 1H NMR (500 MHz, DMSO-d6) δ 7.57 – 7.42 (m, 3H), 7.42 – 7.29 (m, 2H), 7.27 – 7.14 (m, 2H), 7.12 – 6.97 (m, 1H), 6.38 (s, 1H), 3.66 – 3.50 (m, 2H), 3.50 – 3.36 (m, 1H), 3.28 – 3.07 (m, 2H), 3.07 – 2.57 (m, 5H), 2.22 (s, 6H); 13C NMR (126 MHz, DMSO-d6) δ 168.21, 166.29, 161.94 (d, J = 243.9 Hz), 159.94, 141.76 (d, J = 7.8 Hz), 136.62, 136.22, 130.19 (d, J = 8.5 Hz), 129.41, 129.38, 128.12, 126.91, 124.17, 114.75 (d, J = 22.1 Hz), 114.43 (d, J = 20.9 Hz), 108.20, 51.18, 50.21, 49.55, 49.42, 41.27, 39.88, 23.63; HRMS calcd for C25H26N4OF [M + H]+ 417.2093; Found 417.2092. 2-(4,6-Dimethylpyrimidin-2-yl)-5-[(2-ethoxynaphthalen-1-yl)carbonyl]octahydropyrrolo[3,4-c]pyrrole (28). Triturated with diisopropyl ether (1 mL) to give a white powder (66 mg, 46%); mp 178-180 °C; Mixture of atropisomers by NMR (ca. 3:2). Major isomer is tabulated: 1H NMR (500 MHz, DMSO-d6) δ 7.95 (d, J = 9.3 Hz, 1H), 7.92 – 7.85 (m, 1H), 7.56 (d, J = 8.5 Hz, 1H), 7.54 – 7.31 (m, 3H), 6.39 (s, 1H), 4.30 – 4.19 (m, 1H), 4.13 (q, J = 7.0 Hz, 2H), 3.87 – 3.79 (m, 1H), 3.79 – 3.72 (m, 1H), 3.66 – 3.38 (m, 4H), 3.21 – 3.14 (m, 1H), 3.09 – 2.99 (m, 1H), 2.95 – 2.85 (m, 1H), 2.22 (s, 6H), 1.22 (t, J = 7.0 Hz, 3H); All peaks reported: 13C NMR (126 MHz, DMSO-d6) δ 166.43, 165.82, 165.67, 160.20, 160.08, 151.40, 151.06, 130.12, 130.05, 129.99, 128.24, 128.22, 127.99, 127.15, 127.11, 123.81, 123.49, 123.26, 120.95, 120.84, 114.74, 114.40, 108.31, 108.21, 41.29, 40.35, 40.29, 23.61, 14.91, 14.49; HRMS calcd for C25H29N4O2 [M + H]+ 417.2291; Found 417.2280. 2-(4,6-Dimethylpyrimidin-2-yl)-5-{[5-methyl-2-(2H-1,2,3-triazol-2-yl)phenyl] carbonyl}octahydropyrrolo[3,4-c]pyrrole (37). Recrystallized from diethyl ether (1 mL) to give a white crystalline solid (145 mg, 70%); mp 164.5-166 °C; 1H NMR (500 MHz, DMSO-d6) δ 7.93 (s, 2H), 7.74 (d, J = 8.3 Hz, 1H), 7.40 (dd, J = 8.3, 1.9 Hz, 1H), 7.29 (d, J = 1.8 Hz, 1H),


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6.38 (s, 1H), 3.73 (dd, J = 11.5, 7.6 Hz, 1H), 3.65 (dd, J = 12.4, 7.4 Hz, 1H), 3.60 – 3.51 (m, 1H), 3.49 – 3.33 (m, 4H), 3.04 – 2.81 (m, 3H), 2.38 (s, 3H), 2.22 (s, 6H); 13C NMR (126 MHz, DMSO-d6) δ 166.51, 166.42, 160.11, 138.30, 136.11, 133.19, 130.63, 130.32, 128.18, 122.25, 108.20, 51.53, 50.34, 49.78, 49.42, 41.45, 40.18, 23.64, 20.33; HRMS calcd for C22H26N7O [M + H]+ 404.2199; Found 404.2190. Anal. Calcd for C22H25N7O: C, 65.49; H, 6.25; N, 24.30; Found: C, 65.33; H, 6.10; N, 24.03. 2-(4,6-Dimethylpyrimidin-2-yl)-5-{[5-methoxy-2-(1H-1,2,3-triazol-2yl)phenyl]carbonyl}octahydropyrrolo[3,4-c]pyrrole (39). Triturated with diethyl ether (1 mL) to give a white crystalline solid (160 mg, 74%); mp 145.5-147 °C; 1H NMR (500 MHz, DMSOd6) δ 7.90 (s, 2H), 7.75 (d, J = 8.9 Hz, 1H), 7.14 (dd, J = 8.9, 2.9 Hz, 1H), 7.02 (d, J = 2.9 Hz, 1H), 6.38 (s, 1H), 3.84 (s, 3H), 3.72 (dd, J = 11.5, 7.6 Hz, 1H), 3.64 (dd, J = 12.4, 7.5 Hz, 1H), 3.61 – 3.51 (m, 1H), 3.44 (dd, J = 11.5, 5.2 Hz, 1H), 3.42 – 3.33 (m, 3H), 3.05 – 2.83 (m, 3H), 2.22 (s, 6H); 13C NMR (126 MHz, DMSO-d6) δ 166.38, 165.89, 160.09, 158.84, 135.81, 132.34, 128.92, 124.24, 115.28, 112.55, 108.17, 55.73, 51.51, 50.30, 49.74, 49.41, 41.45, 40.17, 23.63; HRMS calcd for C22H26N7O2 [M + H]+ 420.2148; Found 420.2144. Anal. Calcd for C22H25N7O2: C, 62.99; H, 6.01; N, 23.37; Found: C, 62.69; H, 5.78; N, 23.12. 2-{[5-Chloro-2-(2H-1,2,3-triazol-2-yl)phenyl]carbonyl}-5-(4,6-dimethylpyrimidin-2yl)octahydropyrrolo[3,4-c]pyrrole (40). Triturated with diethyl ether (1 mL) to give a white powder (120 mg, 55%); mp 197.5-199.5 °C; 1H NMR (500 MHz, DMSO-d6) δ 8.00 (s, 2H), 7.91 (d, J = 8.7 Hz, 1H), 7.67 (dd, J = 8.7, 2.4 Hz, 1H), 7.64 (d, J = 2.4 Hz, 1H), 6.38 (s, 1H), 3.73 (dd, J = 11.5, 7.6 Hz, 1H), 3.67 (dd, J = 12.4, 7.5 Hz, 1H), 3.62 – 3.52 (m, 1H), 3.46 (dd, J = 11.6, 5.2 Hz, 1H), 3.43 – 3.34 (m, 3H), 3.04 – 2.85 (m, 3H), 2.22 (s, 6H); 13C NMR (126 MHz, DMSO-d6) δ 166.41, 164.82, 160.10, 136.74, 134.06, 132.74, 132.10, 129.91, 127.79, 124.09,


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108.20, 51.49, 50.28, 49.73, 49.51, 41.47, 40.16, 23.63; HRMS calcd for C21H23ClN7O [M + H]+ 424.1653; Found 424.1652. Anal. Calcd for C21H22ClN7O: C, 59.50; H, 5.23; N, 23.13; Found: C, 59.32; H, 5.17; N, 22.70. General Procedure E To a solution of freshly prepared acid chloride (1 equiv) in DCM (2 mL) was amine (150 mg) and N,N-diisopropylethylamine (4 equiv) and the reaction mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (10 mL) and washed with water (5 mL), 1 N sodium hydroxide (5 mL) and water (2 x 5 mL), dried over sodium sulfate, filtered and evaporated. The compounds that were prepared using General Procedure E are as follows. 2-[5-(Biphenyl-2-ylcarbonyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl]quinoxaline (20). Purified by silica gel column chromatography eluting with chloroform:methanol (100:0.75 to 100:2) to give a yellow foam (107 mg, 53%); 1H NMR (500 MHz, DMSO-d6) δ 8.28 (s, 1H), 7.84 (d, J = 8.1 Hz, 1H), 7.64 – 7.55 (m, 2H), 7.50 (td, J = 7.4, 1.5 Hz, 1H), 7.47 – 7.08 (m, 9H), 3.79 – 3.65 (m, 1H), 3.65 – 3.44 (m, 2H), 3.35 – 2.53 (m, 7H); 13C NMR (126 MHz, DMSO-d6) δ 168.53, 150.06, 141.67, 139.50, 137.52, 136.91, 136.59, 135.99, 129.83, 129.33, 128.48, 128.26, 128.07, 127.62, 127.57, 126.90, 125.72, 123.52, 50.85, 50.13, 49.51, 49.20, 41.22; HRMS calcd for C27H25N4O [M + H]+ 421.2028; Found 421.2028. 2-[5-{[2-(1H-Pyrazol-1-yl)phenyl]carbonyl}hexahydropyrrolo[3,4-c]pyrrol-2(1H)yl]quinoxaline (22). Purified by column chromatography (ISCO) eluting with chloroform:methanol (100:0 to100:0.5) to give a a pale yellow foam (85 mg, 41%); 1H NMR (500 MHz, DMSO-d6) δ 8.44 (s, 1H), 8.06 (s, 1H), 7.82 (d, J = 8.1 Hz, 1H), 7.70 – 7.48 (m, 5H),


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7.47 – 7.40 (m, 2H), 7.39 – 7.30 (m, 1H), 6.37 (br s, 1H), 3.92 – 3.80 (m, 1H), 3.76 – 3.60 (m, 2H), 3.59 – 3.41 (m, 3H), 3.41 – 3.23 (m, 1H), 3.12 – 2.75 (m, 3H); 13C NMR (126 MHz, DMSO-d6) δ 166.86, 150.27, 141.69, 140.86, 136.98, 136.04, 135.99, 131.29, 129.85, 129.59, 128.48, 127.88, 127.38, 125.69, 123.50, 123.00, 107.39, 51.21, 50.33, 49.79, 49.28, 41.41, 40.20; HRMS calcd for C24H23N6O [M + H]+ 411.1933; Found 411.1931. 2-(4,6-Dimethylpyrimidin-2-yl)-5-{[4-fluoro-2-(2H-1,2,3-triazol-2-yl)phenyl] carbonyl}octahydropyrrolo[3,4-c]pyrrole (32). Triturated with diethyl ether (1 mL) and recrystallized from acetonitrile (0.6 mL) to give a white crystalline solid (93 mg, 45%); mp 175.2-177.0 °C; 1H NMR (500 MHz, DMSO-d6) δ 8.02 (s, 2H), 7.78 – 7.68 (m, 1H), 7.61 – 7.52 (m, 1H), 7.43 – 7.34 (m, 1H), 6.38 (s, 1H), 3.73 (dd, J = 11.6, 7.6 Hz, 1H), 3.67 (dd, J = 12.4, 7.6 Hz, 1H), 3.61 – 3.51 (m, 1H), 3.50 – 3.33 (m, 4H), 3.04 – 2.82 (m, 3H), 2.22 (s, 6H); 13C NMR (126 MHz, DMSO-d6) δ 166.41, 165.69, 161.82 (d, J = 247.0 Hz), 160.09, 136.90, 136.54 (d, J = 10.6 Hz), 130.36 (d, J = 9.0 Hz), 126.93 (d, J = 3.0 Hz), 115.42 (d, J = 21.4 Hz), 109.47 (d, J = 26.5 Hz), 108.21, 51.53, 50.35, 49.78, 49.54, 41.45, 40.18, 23.64; HRMS calcd for C21H23N7OF [M + H]+ 408.1948; Found 408.1943. Anal. Calcd for C21H22N7OF: C, 61.90; H, 5.44; N, 24.06; Found: C, 61.85; H, 5.31; N, 24.19. 2-(4,6-Dimethylpyrimidin-2-yl)-5-{[5-fluoro-2-(2H-1,2,3-triazol-2-yl)phenyl] carbonyl}octahydropyrrolo[3,4-c]pyrrole (33). Triturated with diethyl ether (1.5 mL) and recrystallized from acetonitrile (0.5 mL) to give a white crystalline solid (50 mg, 24%); mp 141.8-142.4 °C; 1H NMR (500 MHz, DMSO-d6) δ 7.97 (s, 2H), 7.93 – 7.85 (m, 1H), 7.53 – 7.40 (m, 2H), 6.38 (s, 1H), 3.73 (dd, J = 11.5, 7.6 Hz, 1H), 3.65 (dd, J = 12.5, 7.5 Hz, 1H), 3.62 – 3.52 (m, 1H), 3.45 (dd, J = 11.6, 5.2 Hz, 1H), 3.43 – 3.34 (m, 3H), 3.07 – 2.85 (m, 3H), 2.23 (s, 6H); 13C NMR (126 MHz, DMSO-d6) δ 166.40, 164.84, 161.03 (d, J = 247.7 Hz), 160.102,


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136.44, 132.80 (d, J = 7.0 Hz), 131.97, 125.00 (d, J = 9.0 Hz), 116.89 (d, J = 23.3 Hz), 114.95 (d, J = 24.3 Hz), 108.20, 51.47, 50.29, 49.73, 49.49, 41.45, 40.15, 23.62; HRMS calcd for C21H23N7OF [M + H]+ 408.1948; Found 408.1953. Anal. Calcd for C21H22N7OF: C, 61.90; H, 5.44; N, 24.06; Found: C, 61.52; H, 5.30; N, 23.85. (5-(4,6-Dimethylpyrimidin-2-yl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)(3-methyl-2-(2H1,2,3-triazol-2-yl)phenyl)methanone (36). Purified further by column chromatography (ISCO) eluting with chloroform:methanol (100:0 to 100:0.4) to give an off-white foam (90 mg, 32%); 1H NMR (500 MHz, DMSO-d6) δ 7.92 (s, 2H), 7.54 – 7.46 (m, 2H), 7.35 (dd, J = 6.7, 2.3 Hz, 1H), 6.39 (s, 1H), 3.69 (dd, J = 11.5, 6.9 Hz, 1H), 3.59 (dd, J = 11.7, 6.6 Hz, 1H), 3.55 – 3.39 (m, 3H), 3.36 – 3.27 (m, 1H), 3.20 (dd, J = 12.4, 3.5 Hz, 1H), 3.13 (dd, J = 10.8, 4.5 Hz, 1H), 2.96 – 2.85 (m, 2H), 2.24 (s, 6H), 2.10 (s, 3H); 13C NMR (126 MHz, DMSO-d6) δ 166.43, 165.79, 160.12, 135.48, 135.40, 135.36, 134.58, 131.58, 129.60, 124.74, 108.21, 51.83, 50.21, 49.69, 49.18, 41.48, 40.11, 23.65, 17.39; HRMS calcd for C22H26N7O [M + H]+ 404.2199; Found 404.2197. (5-(4,6-Dimethylpyrimidin-2-yl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)(2-methyl-6-(2H1,2,3-triazol-2-yl)phenyl)methanone (38). Purified further by silica gel column chromatography eluting with chloroform:methanol (100:1 to 100:2) to give an off-white foam (134 mg, 64%); Mixture of atropisomers by NMR (ca. 3:2). Major isomer is tabulated: 1H NMR (500 MHz, DMSO-d6) δ 7.88 (s, 2H), 7.66 (d, J = 8.0 Hz, 1H), 7.47 (t, J = 7.8 Hz, 1H), 7.39 (d, J = 7.6 Hz, 1H), 6.38 (s, 1H), 3.80 – 3.60 (m, 2H), 3.52 (dd, J = 11.7, 6.2 Hz, 1H), 3.49 – 3.32 (m, 4H), 3.04 – 2.85 (m, 3H), 2.32 (s, 3H), 2.27 – 2.22 (m, 6H); All peaks reported: 13C NMR (126 MHz, DMSO-d6) δ 166.45, 166.40, 165.89, 165.69, 160.13, 160.11, 136.36, 136.09, 135.52, 135.39, 135.37, 135.29, 130.57, 130.44, 130.10, 130.06, 129.20, 120.04, 119.90, 108.33, 108.15,


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50.98, 50.66, 50.25, 50.16, 49.61, 49.40, 49.21, 41.52, 41.18, 40.18, 23.64, 23.62, 18.51, 18.46; HRMS calcd for C22H26N7O [M + H]+ 404.2199; Found 404.2199. The procedures used to prepare bulk 34 are as follows. (5-(4,6-dimethylpyrimidin-2-yl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)(2-fluoro-6-(2H1,2,3-triazol-2-yl)phenyl)methanone (6s). Step 1 2-Fluoro-6-[1,2,3]triazol-2-yl-benzoic acid (42). To a 2 L, 3-necked, round-bottomed flask equipped with an overhead mechanical stirrer, thermocouple probe, heating mantle, reflux condenser, and nitrogen inlet were added 2-fluoro-6-iodobenzoic acid (127.6 g, 480 mmol), copper iodide (4.57 g, 24 mmol), and Cs2CO3 (312.6 g, 959 mmol). To these solids were added dioxane (640 mL), then water (2.6 mL, 144 mmol), then 1H-1,2,3-triazole (55.6 mL, 959 mmol), and finally trans-1,2-dimethylcyclohexane-1,2-diamine (15.1 mL, 96 mmol). The mixture was then warmed to 60 °C for 30 min, then to 83 °C for 30 min, and then to 100 °C for 3 h. After the 3 h at 100 °C, the mixture was cooled and then 1 L of MTBE and 1 L of water were added. After vigorous mixing, the layers were separated and the bottom aqueous layer was acidified to pH 1.72 with ~148 mL of concentrated hydrochloric acid. The aqueous was then extracted twice with EtOAc. The combined organic layers were dried over Na2SO4, filtered, and concentrated to provide a dark oil. The oil was stirred overnight in EtOAc (450 mL) and the resulting precipitate was removed by filtration. The mother-liquors were concentrated to a brown solid (106.21 g, 75 wt% by quantitative HPLC, 79.7 g, 80%). 1H NMR (400 MHz, DMSO-d6): 8.22 – 8.13 (bs, 2H), 7.84-7.80 (m, 1H), 7.74 – 7.65 (m, 1H), 7.50 – 7.41 (m, 1H). Step 2


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2-Benzyl-5-(4,6-dimethyl-pyrimidin-2-yl)-octahydro-pyrrolo[3,4-c]pyrrole (45) To a 3 L, 3necked, round-bottomed flask equipped with mechanical stirrer, reflux condenser, temperature probe, and nitrogen inlet, was added 2-benzyl-octahydro-pyrrolo[3,4-c]pyrrole (109 g, 538.8 mmol) in DMF (1.6 L). To the resulting solution were added 2-chloro-4,6-methylpyrimidine (76.8 g, 538.8 mmol) and Cs2CO3 (351.1 g, 1.08 mol). The heterogeneous mixture was heated to 100 °C and stirred for 15 h. After cooling to room temperature, the mixture was diluted with EtOAc (1.5 L) and water (1.5 L). The layers were thoroughly mixed and separated. The aqueous layer was extracted with additional EtOAc (1.5 L). The combined organics were dried over sodium sulfate, filtered, and concentrated under reduced pressure to a brown solid (160 g, 96% yield). MS (ESI) mass calcd. for C19H24N4, 308.20; m/z found 309 [M+H]+. 1H-NMR (500 MHz, CDCl3): 7.32 – 7.26 (m, 4H), 7.25 – 7.20 (m, 1H), 6.27 (s, 1H), 3.81 – 3.73 (m, 2H), 3.58 (s, 2H), 3.54 (dd, J = 11.4, 3.5 Hz, 2H), 2.95 – 2.86 (m, 2H), 2.80 – 2.68 (m, 2H), 2.47 – 2.40 (m, 2H), 2.35 – 2.24 (s, 6H). Step 3 2-(4,6-Dimethyl-pyrimidin-2-yl)-octahydro-pyrrolo[3,4-c]pyrrole•HOAc (46). To a 4 L high pressure autoclave equipped with mechanical stirring, temperature probe, heating jacket, and gas inlet were added 5% Pd/C (66.9 g, Johnson Matthey 5R338, 56.8% H2O, 3.45 mol%) and a solution of 2-benzyl-5-(4,6-dimethyl-pyrimidin-2-yl)-octahydro-pyrrolo[3,4-c]pyrrole (160 g, 519 mmol) and HOAc (30 mL, 519 mmol) in ethanol (3.2 L). The mixture was stirred at 50 °C under 50 psi of H2(g) for 4 h. The catalyst was removed and the resulting solution was then concentrated under reduced pressure to provide the desired product as a white solid (144 g, quantitative yield) as the HOAc salt. MS (ESI): mass calcd. for C12H18N4, 218.15; m/z found


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219 [M+H]+. 1H-NMR (CDCl3, 400 MHz): 6.30 (s, 1H), 3.79 – 3.59 (m, 4H), 3.39 (m, 2H), 3.09 – 2.88 (m, 4H), 2.29 (s, 6H), 1.93 (s, 3H). Step 4 (5-(4,6-dimethylpyrimidin-2-yl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)(2-fluoro-6-(2H1,2,3-triazol-2-yl)phenyl)methanone (34). To a 3-necked, 3 L, round-bottomed flask equipped with a nitrogen line, temperature probe, heating mantle, reflux condenser, mechanical stirrer, and 1 N aq. NaOH scrubber were added 2-fluoro-6-[1,2,3]triazol-2-yl-benzoic acid (42), (120.98 g, 75 wt%, 90.74 g actual, 438 mmol) and toluene (1 L). The mixture was warmed to 50 °C for 1 h with stirring. The mixture was then cooled to 25 °C and thionyl chloride (47.9 mL, 657 mmol) was added. The mixture was warmed back to 50 °C and held for 1 h. During this time, in a separate 5 L jacketed reactor equipped with a mechanical stirrer and temperature probe were added toluene (600 mL), aqueous sodium carbonate (185.7 g, 1.75 mol in 1.6 L water), and 2(4,6-dimethyl-pyrimidin-2-yl)-octahydro-pyrrolo[3,4-c]pyrrole•HOAc (46), (122 g, 438 mmol). This biphasic mixture was cooled to 0 °C. After cooling to 0 °C, the original slurry was poured through a filter and over the stirring biphasic mixture of amine and aqueous sodium carbonate. The mixture was allowed to warm to room temperature. After 2 h, additional 2-(4,6-dimethylpyrimidin-2-yl)-octahydro-pyrrolo[3,4-c]pyrrole•HOAc (4 g, 14 mmol) was added and the mixture was stirred for 30 additional minutes. At the end of this period, the layers were separated and 100 mL of methanol were added to the organic layer. The organic layer was dried over MgSO4, filtered, and concentrated to a white solid. This solid was taken up in ethanol (1.4 L) and warmed to 77 °C. The mixture was then cooled to 55 °C and seeded with previously crystallized material. (Note: The seeds were generated from slurrying the initial product in 2propanol at room temperature [100 mg/mL]). The mixture was cooled to room temperature at a


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rate of 5 °C per hour. After stirring at room temperature for 14 h, the mixture was filtered and dried to provide the final product as a white crystalline solid (136.84 g, 74%). 1H NMR (400 MHz, CDCl3): 7.88 – 7.78 (m, 1.78H), 7.75 – 7.69 (s, 1.22H), 7.51 – 7.43 (m, 1H), 7.17 – 7.11 (m, 1H), 6.30 – 6.28 (m, 1H), 4.03 – 3.48 (m, 7H), 3.29 – 3.21 (m, 1H), 3.15 – 2.92 (m, 2H), 2.30 (s, 6H). 13C NMR All peaks reported (151 MHz, DMSO-d6) 171.73, 171.67, 166.62, 166.50, 165.40, 165.37, 164.25, 162.62, 162.50, 142.32, 142.24, 142.06, 141.97, 141.61, 141.57, 136.69, 124.21, 124.11, 124.05, 120.77, 120.61, 113.59, 113.46, 56.35, 56.15, 55.85, 55.53, 55.30, 54.97, 54.94, 54.84, 46.64, 46.43, 45.52, 45.46, 45.19, 45.05, 44.91, 44.77, 44.63, 44.49, 44.35, 28.91, 28.89. MS (ESI) mass calcd for C21H22FN7O, 407.19; m/z found, 408 [M+H]+. Anal. calcd. for C21H22FN7O C, 61.90, H, 5.44, N, 24.06; found C, 61.83, H, 5.42, N, 24.08. DMPK. General methods. A 10 mM compound stock solution in DMSO further diluted in acetonitrile:water to yield a secondary 1 mM solution was used in all in vitro assays. Pooled human liver microsomes of mixed gender, mouse, rat, beagle dog, and cynomolgus monkey of male gender were purchased from BD Gentest (Woburn, MA). Plasma from various species was purchased from Bioreclamation, Inc. (Westbury, NY). All other commercially available reagents and solvents were either analytical or HPLC grade. In vitro and PK samples were analyzed by LC-MS/MS in the Multiple Reaction Monitoring (MRM) scan mode with electrospray ionization (ESI). Caco-2 permeability. Bi-directional permeability (A to B and B to A, pH7.4/pH7.4) was assessed in a subclone of the Caco-2 cell line, TC7, by Cerep at a compound concentration of 10 µM using the standard Cerep protocol.


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Plasma Protein Binding. The stock solution of compound was spiked into blank plasma to yield 0.1, 1 or 10 µM final concentration. Equilibrium dialysis was performed using the RED Device (Thermo Scientific, Rockford, IL) consisting of Teflon base plate and RED Device inserts comprised of two (sample and buffer) side-by-side chambers separated by dialysis membrane (MWCO~8,000). A 300 µL aliquot of plasma spiked with a compound in triplicate was placed into the sample chamber and 500 µL of dialysis buffer was loaded into the buffer chamber. The dialysis unit was covered with sealing tape and incubated in a 37 °C orbital shaking incubator at 100 rpm for 6 h. After a 6-hour dialysis, 10 µL of plasma sample was diluted with 90 µL of PBS buffer, and 90 µL of each buffer sample was mixed with 10 µL of blank plasma in a 96 well plate and extracted with 150 µL of acetonitrile to precipitate proteins. The supernatant was analyzed by LC-MS/MS. The preliminary (Tier 1) assay was conducted at a compound concentration of 1 µM using a 2-point (0 and 1 µM) calibration curve. The definitive plasma protein binding for compound 34 was conducted in 5 species at 3 different concentrations (0.1, 1 or 10 µM) using the full calibration curve and 3 sets of quality control samples. Calibration curve samples were prepared by spiking appropriate stock solutions into a mixture of blank plasma and PBS buffer (1:9, v/v). Brain Tissue Binding. Brain tissue binding was assessed by an equilibrium dialysis technique similar to the procedure described for plasma protein binding. Rat brain tissue homogenate prepared in PBS buffer, pH7.4, (1:10, w/v) was spiked with compound DMSO stock solution to yield a final concentration of 5 µM. The dialysis was carried out in a shaking incubator at 37 °C for 5 h. After incubation, 25 µl homogenate or 50 µl buffer were extracted with 50 µl DMSO and 300 µl acetonitrile and analyzed by LC-MS/MS using the calibration curves across an appropriate concentration range and quality control samples. All determinations were conducted


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in triplicate. The apparent unbound fraction (fu,app) was determined as

f

u , app

=

[ A]buffer [ A] hom ogenate

where

[A]homogenate is the concentration measured in the homogenate and [A]buffer the concentration measured in the buffer. The unbound fraction in undiluted brain was calculated as

f

u ,brain

fu

= D+

fu

app

app

− D×

percentage

compound

% BTB = (1 −

f

u ,brain

fu

, where D is the dilution factor of 10. Subsequently, the app

bound

to

brain

tissue

(%BTB)

was

determined

as

) × 100% .

Stability in Liver Microsomes. Stability in liver microsomes from human, mouse, rat, dog, or monkey was assessed over a time course of 1 h. The incubation mixture consisted of 0.5 mg/mL microsomal protein, 1 µM compound, 1.8 mM MgCl2, and 2 mM NADPH in 0.1 M potassium phosphate buffer, pH 7.4. After incubation for 5, 10, 20, 40 and 60 min at 37 °C, the reaction was terminated by the addition of 5 volumes of acetonitrile containing internal standard (5,5diphenylhydantoin). The samples were analyzed by LC-MS/MS and the percent remaining at various time points was calculated by dividing the observed analyte/internal standard peak ratio by the ratio at the 0 time point. The half-life (t1/2) was calculated as t1/2 = ln(2)/-k, where k is the slope of the linear regression from natural log percent remaining versus time. In vitro hepatic clearance (CL) was calculated as CL = Q*CLint/(Q+ CLint), where Q - hepatic blood flow in mL/min/kg: mouse - 90, rat - 55, dog - 31, monkey - 44, human - 21; CLint - intrinsic microsomal clearance determined as CLint = -k * microsomal protein conc –1 * A * N, where A - microsomal protein yield (45 mg) per 1 g of liver, N - 25.7, 30, 32, 40 and 87.5 g of liver per kg of body weight for human, monkey, dog, rat and mouse, respectively. Extraction ratio (ER) was calculated as ER = CL/Q.


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CYP Inhibition. The potential to inhibit various isoforms of human Cytochrome P450 (CYP1A2, 2C8, 2C9, 2C19, 2D6, and 3A4) was investigated by incubating human liver microsomes (0.2 mg/mL) with a ‘cocktail’ of CYP probe substrates (phenacetin for CYP1A2, paclitaxel for CYP2C8, diclofenac for CYP2C9, S-mephenytoin for CYP2C19, dextromethorphan for 2D6, and midazolam for CYP3A4) in the presence of compound at concentrations ranging from 0 (vehicle) to 10 µM. Metabolite formation by probe substrates was quantified by LC-MS/MS. The percent inhibition was measured as the ratio of metabolite peak in the presence and absence of compound*100. The IC50 values were calculated from the percent inhibition data relative to vehicle. Pharmacokinetic Studies. Single dose pharmacokinetic studies in preclinical species (male Balb/c mice, Sprague Dawley rats, beagle dogs or cynomolgus monkeys) were conducted following i.v. (1 mg/kg) and p.o. (5 mg/kg) administration as a solution in 20% hydroxypropylbeta-cyclodextrin (HP-β-CD). Blood was sampled at pre-dose and at 0.033 (i.v.), 0.083 (i.v.), 0.25, 0.5, 1, 2, 4, 6, 8, and 24 h post dose. In dogs, instead of 6 and 8 h time points blood was drawn at 7 h. Plasma concentrations were quantitated by LC-MS/MS. Pharmacokinetic parameters were derived from noncompartmental analysis of the plasma concentration vs. time data using WinNonlin software (Pharsight, Palo Alto, CA). Developability Assay Protocols HPLC. An Agilent model 1100 equipped with a diode array detector was used for chromatographic quantification. A linear gradient method from 5% to 100% solvent B (organic phase) over 15 minutes at a rate of 1.0mL/min was used to separate the compound and its impurities/degradants on a 150 x 4.6 mm Phenomenex Luna C18 (2) 5µ 100Å column,


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maintained at 40 °C. The aqueous mobile phase consisted of a 0.05% TFA solution in water while the organic phase contained 0.05% TFA in acetonitrile. Detection was done by UV at 240 nm. Oxidation stability studies. Oxidative stability in solution was assessed in the presence of 0.1% H2O2 and 5 mM AIBN, a free-radical forming agent. Solutions (0.1 mg/mL) in 1:1 Methanol/water containing either H2O2 or AIBN were incubated for 24 hours at room temperature or at 40 ºC, respectively and then assayed by HPLC against external standards and controls. Photostability studies. The light stability in 50% aqueous Acetonitrile (0.1 mg/mL) was investigated under ICH guideline, option 2 (cool white fluorescent light: 1.2 million lux-hours and near UV: 200 W-h/m2) using a Caron Photo-stability Chamber Model # 6540-1. The samples were analyzed by HPLC, against external standards. Powder X-ray Diffractometry (PXRD). Philips X’PERT PRO with X'Celerator Cu detector equipped with a real time multiple strips X-ray detection technology was used to obtain x-ray powder patterns of the various batches of JNJ-42847922. The samples were scanned from 4o to 40o 2θ, at a step size 0.0167o 2θ and a time per step of 29.8450 seconds. The tube voltage and current were 45 kV and 40 mA, respectively. The samples were placed onto zero background holders and analyzed on a spinning stage. Thermogravimetric Analysis (TGA). The analyses were performed on a TA Instruments model TGA Q 50 and Q 500 under a nitrogen purge. The samples were placed in aluminum pans and scanned at a rate of 10 ºC /min up to 400 ºC.


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Differential Scanning Calorimetry (DSC). DSC scans were recorded on a TA Instruments model DCS Q 100. The samples, placed in hermetic DSC pans punctured with a single hole in the lid, were scanned at a rate of 10 ºC /min under a nitrogen purge. Moisture Sorption Analysis. Moisture isotherms were recorded on a Hiden Isochema model IGAsorp Moisture Sorption Analyzer. After weight stabilization at 40% relative humidity (RH), the sample was introduced to increasing amounts of humidity at 10% RH intervals up to 90% RH. During the desorption cycle, the sample was brought to 0%RH in 10%RH increments. Experiments were conducted isothermally at 25 ºC. Solubility in Aqueous Systems. Solubility in 30 mM Phosphate buffers (pH 2 and pH 7), simulated gastric (SGF: 0.2% NaCl in 0.1-N HCl; pH 1.2) and intestinal fluids (FasSIF: 0.029M Phosphate buffer, 5-mM Sodium Taurocholate, and 1.5-mM Lecithin; pH 6.8) was investigated. Compound was dissolved in DMSO solutions at a concentration of 10 mM weas used for the solubility experiment. DMSO solutions ( 20 µL) are dispensed in 96-well plates and the solvent is removed by evaporation using a Caliper TurboVap 96 set at 30°C and a flow rate of 40 Fh. Buffers (400 µL) of interest are added to the residual solids and the resulting mixtures are stirred at room temperature for 3 days using magnetic stir bars. The samples are then filtered using an AcroPrep 1 mL 96 Filter Plate and the supernatant is analyzed for compound concentration, against external standards. Solid state stability: The solid active pharmaceutical ingredient (API) was stored under ambient laboratory conditions and at 80 °C/ambient relative humidity (RH) in closed vial configuration and at 40 °C/75%RH in open vial configuration. At various time intervals, the solid material was sampled and analyzed by HPLC and PXRD to assess its chemical and physical stability.


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Oxidative Stability in Solution. Oxidative stability of the molecule in solution was assessed in the presence of 0.1% H2O2 and 5 mM AIBN, a free-radical forming agent. Solutions of JNJ54717793 in 1:1 Methanol/water containing either H2O2 or AIBN were incubated for 24 hours at room temperature or at 40 ºC, respectively and then assayed by HPLC against external standards and controls. Photolytic Stability in Solution. Solution photostability (0.1 mg/mL) in 1:1 acetonitrile:water mixture was assessed using the Caron Chamber Model # 6540-1 (ICH Guideline, Option 1). The controls were stored in aluminum-wrapped vials. The samples were then assayed by HPLC against external standards. Chemical Stability as a Function of pH. The chemical stability as a function of pH at room temperature and 60 ºC was assessed from pH 2 (30 mM Phosphate), pH 4 (30 mM Acetate), pH 7.4 (30 mM Phosphate), and pH 10 (30-mM Carbonate) solutions after one and four weeks of storage. The samples were assayed by HPLC against external standards

ASSOCIATED CONTENT Supporting Information. This material is available free of charge via the Internet at http://pubs.acs.org. AUTHOR INFORMATION Corresponding Author * (M.A.L.) e-mail: [email protected]. Tel: 1-858-450-2302. Author Contributions


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The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript. ACKNOWLEDGMENT The authors gratefully acknowledge the chemistry team at BioBlocks, Inc for the preparation and characterization of compounds 11-15, 16-33 and 35-40. ABBREVIATIONS DMPK, drug metabolism and pharmacokinetics, SAR, structure-activity relationship, MW, molecular weight, p.o, per os, OX2R, orexin-2 receptor, OX1R, orexin-1 receptor, CL, clearance, Vss, volume of distribution at steady state, t1/2, half-life, %F, percent bioavailability, RLM, rat liver microsomes, HLM, human liver microsomes, CYP, cytochrome P450, R.O., receptor occupancy, PPB plasma protein binding, NREM, non-rapid eye movement, REM, rapid eye movement, EEG, electroencephalogram, GLP, good laboratory practices, PK, pharmacokinetics, SGF simulated gastric fluid, SIF, simulated intestinal fluid. REFERENCES (1)

de Lecea, L.; Kilduff, T. S.; Peyron, C.; Gao, X.; Foye, P. E.; Danielson, P. E.;

Fukuhara, C.; Battenberg, E. L.; Gautvik, V. T.; Bartlett, F. S. 2nd, Frankel, W. N.; van den Pol, A. N.; Bloom, F. E.; Gautvik, K. M.; Sutcliffe, J. G. The hypocretins: hypothalamusspecific peptides with neuroexcitatory activity. Proc. Natl. Acad. Sci. 1998, 95:322-327. (2)

Chemelli, R. M.; Willie, J. T.; Sinton, C. M.; Elmquist, J. K.; Scammell, T.; Lee,

C.; Richardson, J. A.; Williams, S. C.; Xiong, Y.; Kisanuki, Y.; Fitch, T. E.; Nakazato, M.; Hammer, R. E.; Saper, C. B.; Yanagisawa, M. Narcolepsy in orexin knockout mice: molecular genetics of sleep regulation. Cell 1999, 98:437-451. (3)

Peyron, C.; Faraco, J.; Rogers, W.; Ripley, B.; Overeem, S.; Charnay, Y.;

Nevsimalova, S.; Aldrich, M.; Reynolds, D.; Albin, R.; Li, R.; Hungs, M.; Pedrazzoli, M.;


56

Page 57 of 61

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


Padigaru, M.; Ku-cherlapati, M.; Fan, J.; Maki, R.; Lammers, G. J.; Bouras, C.; Kucherlapati, R.; Nishino, S.; Mignot, E. A mutation in a case of early onset narcolepsy and a generalized absence of hypocretin peptides in human narcoleptic brains. Nat. Med. 2000, 6:991-997. (4)

For reviews on orexin antagonists see: (a) Gotter, A. L.; Roecker, A. J.

Hargreaves, R.; Coleman, P. J.; Winrow, C. J.; Renger, J. J. Orexin receptors as therapeutic drug targets Prog. Brain Res. 2012, 198, 163-188. (b) Gatfield, J.; Brisbare-Roch, C.; Jenck, F.; Boss, C. Orexin receptor Antagonists: A New Concept in CNS Disorders Chem. Med. Chem. 2010, 5, 1197-1214, (c) Mieda, M.; Sakurai, T. Orexin (Hypocretin) Receptor Agonists and Antagonists for Treatment of Sleep Disorders: Rationale for Development and Current Status. CNS Drugs, 2013, 27:83-90, (d) Winrow, C. J.; Renger, J.J. Discovery and development of orexin receptor antagonists as therapeutics for insomnia. 2014. Br. J. Pharmacol. 171:283-293,(e) Kuduk, S. D.; Winrow, C. J.; Coleman, P. J. Orexin Receptor Antagonists in Development for Insomnia and CNS Disorders. Ann. Rep. Med. Chem., 2013,48, 73-87. (5)

Hoever, P.; de Haas, S.; Winkler, J.; Schoemaker, R. C.; Chiossi, E.; van Gerven,

J.; Dingemanse, J. Orexin Receptor Antagonism, a New Sleep-Promoting Paradigm: An Ascending Single-Dose Study With Almorexant Clin. Pharmacol. Ther. 2010, 87, 593-600. (6)

Cox, C. D.; Breslin, M. J.; Whitman, D. B.; Schreier, J. D.; McGaughey, G. B.;

Bogusky, M. J.; Roecker, A. J.; Mercer, S. P.; Bednar, R. A.; Lemaire, W.; Bruno, J. G.; Reiss, D. R.; Harrell, C. M.; Murphy, K. L.; Garson, S. L.; Doran, S. M.; Prueksaritanont, T.; Anderson, W. B.; Tang, C.; Roller, S.; Cabalu, T. D.; Cui, D.; Hartman, G. D.; Young, S. D.; Koblan, K. S.; Winrow, C. J.; Renger, J. J.; Coleman, P. J. Discovery of the Dual Orexin Receptor Antagonist [(7R)-4-(5-Chloro-1,3-benzoxazol-2-yl)-7-methyl-1,4diazepan-1-yl][5-methyl-2-(2H-1,2,3-triazol-2-yl)phenyl]methanone (MK-4305) for the Treatment of Insomnia J. Med. Chem. 2010, 53, 5320-5332. (7)

Yang, L. P. H. Suvorexant: First Global Approval. Drugs, 2014, 74:1817-1822.


57


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(8)

Page 58 of 61

For a review describing selective orexin receptor antagonist see: Lebold, T. P.;

Bonaventure, P.; Shireman, B. T. Selective Orexin receptor Antagonists. Bioorg. Med. Chem. Lett., 2013, 23 (17). 4761-4769. (9)

Dugovic, C.: Shelton, J. E.; Aluisio, L. E.; Fraser, I. C.; Jiang, X.; Sutton S .W.;

Bonaventure, P.; Yun, S.; Li, X.; Lord, B.; Dvorak, C.A.; Carruthers, N. I.; and Lovenberg, T. W. Blockade of orexin-1 receptors attenuates orexin-2 receptor antagonism-induced sleep promotion in the rat. J. Pharmacol. Exp. Ther. 2009, 330, 142-151. (10)

Dugovic, C.: Shelton, J. E.; Sujin, Y.; Bonaventure, P.; Shireman, B. T.;

Lovenberg, T. W. Orexin-1 receptor blockade dysregulates REM sleep in the presence of orexin-2 receptor antagonism Front. Neurosci., 2014, 8, article 28. (11)

Reference 10 and references therein.

(12)

Morairty, S. R.; Revel, F. G.; Malherbe, P.; Moreau, J. L.; Valladao, D.;

Wettstein, J. G.; Kilduff, T. S.; Edilio, B. Dual hypocretin receptor antagonism is more effective for sleep promotion than antagonism of either receptor alone. PLoS ONE, 2012, 7(7), e39131. (13)

McAtee, L. C.; Sutton, S. W.; Rudolph, D. A.; Li, X.; Aluisio, L. E.; Phuong, V.

K.; Dvorak, C. A.; Lovenberg, T. W.; Carruthers, N. I.; Jones, T. K. Novel substituted 4phenyl-[1,3]dioxanes: potent and selective orexin receptor 2 (OX2R) antagonists Bioorg. Med. Chem. Lett. 2004, 14 (16), 4225-4229. (14)

Whitman, D. B.; Cox, C. D.; Breslin, M. J.; Brashear, K. M.; Schreier, J. D.;

Bogusky, M. J.; Bednar, R. A.; Lemaire, W.; Bruno, J. G.; Hartman, G. D.; Reiss, D. R.; Harrell, C. M.; Kraus, R. L.; Li, Y.; Garson, S. L.; Doran, S. M.; Prueksaritanont, T.; Li, C.; Winrow, C. J.; Koblan, K. S.; Renger, J. J.; Coleman, P. J. Discovery of a Potent, CNSPenetrant Orexin Receptor Antagonist Based on an N,N-Disubstituted-1,4-diazepane Scaffold that Promotes Sleep in Rats Chem. Med. Chem. 2009, 4 (7), 1069-1074. (15)

Roecker, A. J.; Mercer, S. P.; Schreier, J. D.; Cox, C. D.; Fraley, M. E.; Steen, J.

T.; Lemaire, W.; Bruno, J. G.; Harrell, C. M.; Garson, S. L.; Gotter, A. L.; Fox, S. V.;


58

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Stevens, J.; Tannenbaum, P. L.; Prueksaritanont, T.; Cabalu, T. D.; Cui, D.; Stellabott, J.; Hartman, G. D.; Young, S. D.; Winrow, C. J.; Renger, J. J.; Coleman, P. J. Discovery of 5’’Chloro-N-[(5,6-dimethoxypyridin-2-yl)methyl]-2,2’:5’,3’’-terpyridine-3’-carboxamide (MK-1064): A Selective Orexin 2 Receptor Antagonist (2-SORA) for the Treatment of Insomnia Chem. Med. Chem. 2014, 9 (2), 311-332. (16)

Roecker, A. J.; Reger, T. S.; Mattern, M. C.; Mercer, S. P.; Bergman, J. M.;

Schreier, J. D.; Cube, R. V.; Cox, C. D.; Li, D.; Lemaire, W.; Bruno, J. G.; Harrell, C. M.; Garson, S. L. ; Gotter, A. L.; Fox, S. V.; Stevens, J.; Tannenbaum, P. L.; Prueksaritanont, T.; Cabalu, T. D.; Cui, D.; Stellabott, J.; Hartman, G. D.; Young, S. D.; Winrow, C. J.; Renger, J. J.; Coleman, P. J. Discovery of MK-3697: A selective orexin 2 receptor antagonist (2-SORA) for the treatment of insomnia Bioorg. Med. Chem. Lett. 2014, 24 (20), 4884-4890. (17)

Betschart, C.;Hintermann, S.; Behnke, D.; Cotesta, S.; Fendt, M.; Gee, C. E.;

Jacobson, L. H.; Laue, G.; Ofner, S.; Chaudhari, V.; Badiger, S.; Pandit, C.; Wagner, J.; Hoyer, D. Identification of a Novel Series of Orexin Receptor Antagonists with a Distinct Effect on Sleep Architecture for the Treatment of Insomnia J. Med. Chem. 2013, 56 (19), 7590-7607. (18)

Raheem, I. T.; Breslin, M. J.; Bruno, J.; Cabula, T. D.; Cooke, A.; Cox, C. D.;

Cui, D.; Garson, S.; Gotter, A. L.; Fox, S. V.; Harrell, C. M.; Kuduk, S. D.; Lemaire, W.; Preuksaritanont, T.; Renger, J. J.; Stump, C.; Tannenbaum, P. L.; Williams, P. D.; Winrow, C. J.; Coleman, P. J. Discovery of piperidine ethers as selective orexin receptor antagonist (SORAs) inspired by filorexant. Bioorg. Med. Chem. Lett. 2015, 25, 444-450. (19)

Roecker, A. J.; Mercer, S. P. Bergman, J. M.; Gilbert, K. F.; Kuduk, S. D.;

Harrell, C. M.; Garson, S.; Fox, S. V.; Gotter, A. L.; Tannenbaum, P. L.; Preuksaritanont, T.; Cabalu, T. D.;Cui, D.; Lemaire, W.; Winrow, C. J.; Renger, J. J.; Coleman, P. J. Discovery of diazepane amide DORAs and 2-SORAs enabled by exploration of isosteric quinazoline replacements. Bioorg. Med. Chem. Lett. 2015, in press, http://dx.doi.org/10.1016/j.bmcl.2014.12.081.


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(20)

Page 60 of 61

Branstetter, B. J.; Letavic, M. A.; Ly, K. S.; Rudolph, D. A.; Savall, B. M.; Shah,

C. R.; Shireman, B. T. PCT Int. Appl. (2011), WO 2011050202 A1 Apr 28, 2011. (21)

Chai, W.; Letavic, M. A.; Ly, K. S.; Pippel, D. J.; Rudolph, D. A.; Sappey, K. C.;

Savall, B. M.; Shah, C. R.; Shireman, B. T.; Soyode-Johnson, A., Stocking, E. M.; Swanson, D. M. PCT Int. Appl. (2011), WO 2011050198 A1 Apr 28, 2011. (22)

The detailed procedures used to measure OX2R and OX1R Ki’s can be found in;

Bonaventure, P.; Yun, S.; Johnson, P. L.; Shekar, A.; Fitz, S. D.; Shireman, B. T.; Lebold, T. P.; Nepomuceno, D.; Lord, B.; Wennerholm, M.; Shelton, J.; Carruthers, N.; Lovenberg, T.; Dugovic, C. A Selective Orexin-1 Receptor Antagonist Attenuates Stress-Induced Hyprarousal without Hypnotic Effects J. Pharmacol. Exp. Ther., 2015, 352(3), 590-601. (23)

The detailed procedures used for conducting these experiments can be found in

reference 9. (24)

Bonaventure, P.; Shelton, J.; Yun, S.; Nepomuceno, D.; Sutton, S.; Aluisio, L.;

Frazer, I.; Lord, B.; Shoblock, J.; Welty, N.; Chaplan, S. R.; Aguilar, Z.; Halter, R.; Ndifor, A.; Koudriakova, T.; Rizzolio, M.; Letavic, M.; Carruthers, N.; Lovenberg, T.; Dugovic, C. Characterization of JNJ-42847922, a Selective Orexin-2 Receptor Antagonist, as a Clinical Candidate for the Treatment of Insomnia submitted for publication in J. Pharmacol. Exp. Ther., in revision, May 2015. (25)

Lave, T. H.; Dupin, S.; Schmitt, C.; Chou, R. C.; Jaeck, D.; Coassolo, P.

Integration of in Vitro Data into Allometric Scaling To Predict Hepatic Metabolic Clearance in Man: Application to 10 Extensively Metabolized Drugs J. Pharm. Sci. 1997 86(5), 584590.

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