Exploiting polarity and chirality to probe the Hsp90 C-terminus

Bioorganic & Medicinal Chemistry xxx (2018) xxx–xxx

Contents lists available at ScienceDirect

Bioorganic & Medicinal Chemistry

journal homepage: w ww.elsevier.com/locate/bmc

Exploiting polarity and chirality to probe the Hsp90 C-terminus

⇑

Leah K. Forsberg, Rachel E. Davis, Virangika K. Wimalasena, Brian S. J. Blagg

Department of Chemistry and Biochemistry, University of Notre Dame, 305 McCourtney Hall, Notre Dame, IN 46556 USA

a r t i c l e i n f o

a b s t r a c t

Article history:

Inhibition of the Hsp90 C-terminus is an attractive therapeutic approach for the treatment of cancer.

Novobiocin, the ﬁrst Hsp90 C-terminal inhibitor identiﬁed, contains a synthetically complex noviose

sugar that has limited the generation of structure-activity relationships for this region of the molecule.

The work described herein utilizes various ring systems as noviose surrogates to explore the size and nat-

ure of the surrounding binding pocket.

Received 23 February 2018

Revised 3 April 2018

Accepted 12 April 2018

Available online xxxx

1. Introduction

inhibitor.^12,13In contrast to N-terminal inhibitors, treatment of

cancer cells with novobiocin did not induce the heat shock

Molecular chaperones maintain cellular protein homeostasis by

orchestrating the proper folding and degradation of cellular pro-

teins. The 90 kDa heat shock protein (Hsp90) is a molecular chap-

erone that is responsible for the maturation and folding of more

than 300 protein substrates (clients).^1,2Hsp90-dependent client

proteins are associated with a variety of disease states, including

neurodegenerative disorders, infectious diseases, and cancer.^3,4

Hsp90 exists as a homodimer and contains an N-terminal ATP-

binding site that catalyzes the hydrolysis of ATP to provide the req-

uisite energy for the protein folding cycle; a middle domain that

maintains interactions with co-chaperones and substrates; and a

C-terminal dimerization domain that contains an MEEVD motif

that is critical for interactions with various co-chaperones.⁵

Hsp90 client proteins are associated with all 10 hallmarks of can-

cer, making it an ideal drug target for the development of cancer

therapeutics.^6–10Inhibitors of the N-terminal ATP-binding site

have been pursued clinically for the treatment of cancer and 17

small molecules have been evaluated in clinical trials.^8,11Unfortu-

nately, classical N-terminal inhibitors induce the pro-survival heat

shock response at the same concentration required to induce the

degradation of client proteins. As a result, toxicities and dosing dif-

ﬁculties have been observed with most of these investigational

new drugs. Inhibitors of the Hsp90 C-terminal dimerization

domain represent an alternative approach to overcome these detri-

ments, as these compounds can segregate client protein degrada-

tion from induction of the heat shock response, which may

overcome the dosing and scheduling issues observed in the clinic.

The ﬁrst small molecule shown to interact with the Hsp90

C-terminus was novobiocin (1, Fig. 1), a clinically used DNA gyrase

response at concentrations that led to client protein degradation.¹²

Initial structure-activity relationship studies revealed that removal

of the 4-hydroxyl and 3⁰-carbamate moieties on novobiocin abol-

ished DNA gyrase inhibitory activity.¹⁴Removal of the prenyl moi-

ety and further optimization of the benzamide side chain led to

compound 2, an inhibitor with 100-fold greater potency than

novobiocin (Fig. 1).¹⁵Unfortunately, the synthetic complexity asso-

ciated with preparation of the noviose sugar limited the elucida-

tion of structure-activity relationships (SAR) for this part of the

molecule and prompted efforts to identify synthetically accessible

surrogates.^16–20Ultimately, it was found that replacement of

noviose with piperidine (3 and 4, Fig. 1) maintains biological activ-

ity and provides a handle to further probe structure-activity rela-

tionships.¹⁷The work described in this article focuses on the

preparation of noviose surrogates, the biological activity mani-

fested by these analogs, and the establishment of structure-activity

relationships for this scaffold.

2. Results and discussion

Prior investigations to modify the piperidine ring focused on the

attachment of aliphatic appendages.^17–20However, it has been

shown that small molecules that contain hydrophilic moieties also

bind to the C-terminal nucleotide-binding site, such as GTP, ATP,

and others (Fig. 2).²¹Since the novobiocin-binding site appears to

overlap with the C-terminal nucleotide-binding domain, modiﬁca-

tions to 3 and 4 were made that included variable length, bulk, and

polarity to probe the novobiocin-binding site and to elucidate

structure-activity relationships.

Synthesis of these analogs began via the reaction of

carboxybenzyl-protected glycine methyl ester (5, Scheme 1) with

Bredereck’s reagent to give enamine 6, which was condensed with

⇑

Corresponding author.

E-mail address: bblagg@nd.edu (Brian S. J. Blagg).

https://doi.org/10.1016/j.bmc.2018.04.028

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L.K. Forsberg et al. / Bioorganic & Medicinal Chemistry xxx (2018) xxx–xxx

2-methylresorcinol to yield coumarin 7. Acetylation of the phenol

and subsequent hydrogenolysis gave aniline 8, which was then

coupled with biphenyl acid chloride 9 before solvolysis of the ester

to unmask phenol 10. Mitsunobu etheriﬁcation with 1-boc-3- and

4-hydroxypiperidine and subsequent removal of the carbamate

gave piperidines 3 and 4, which after microwave-assisted S_N2 cou-

pling with alkyl bromides, provided the ﬁnal products, 11a–g and

12a–g.

Once synthesized, the anti-proliferative activity manifested by

these piperidine analogs was evaluated against the MCF7 and

SKBR3 breast cancer cell lines via an MTS assay (Table 1). In

general, the 3-piperidines exhibited a variation in potency that

correlated with substituent identity, while the 4-piperidines were

relatively equipotent, and produced a ﬂat SAR. For example,

the 3-substituted alcohol derivatives were more potent than the

methyl ethers (11a/c vs 11b/d), a trend which is absent in the 4-

substitued series, 12a–d. In addition, the bulkier carbamate, ester,

and sulfonamide substitutions were not tolerated at the 3-position

(11e–g), but when placed at the 4-position resulted in only a mod-

erate loss in potency (12e–g). Moreover, the 4-piperidine analogs

(12a–g) were, in general, more potent than the corresponding

3-piperidine analogs (11a–g), particularly in the MCF7 cell line.

However, none of these analogs were more potent than the parent

compounds, 3 and 4. Together, these data suggest that the

3-piperidine projects substituents into a spatially restricted area

of the binding pocket, while the 4-piperidine substituents do not.

The sulfonamides 11g and 12g, which serve as phosphate isosteres,

also manifested a reduction in potency, suggesting that the phos-

phate binding site may not overlap with this region of the sugar.

In light of these results, we envisioned that replacement of the

piperidine with a cyclohexanol ring, would reduce the size of alco-

hol 11a and produce compounds that reside within the constrained

binding pocket surrounding the 3-position. Additionally, we

hypothesized that removal of the hydrogen bond donor/acceptor

from the ring system, by exchanging the piperidine for a cyclo-

hexylamine, would facilitate electrostatic interactions and increase

ﬂexibility. Such analogs were easily accessible via Mitsunobu

etheriﬁcation of phenol 10 with the corresponding cyclohexanedi-

ols or aminocyclohexanols (Scheme 2).

Fig. 1. Hsp90 C-terminal inhibitors.

As illustrated in Table 2, replacement of the piperidine with a

cyclohexanol or cyclohexylamine did not produce compounds that

exhibited greater potency. Instead, incorporation of an exocyclic

amine or alcohol at the 3-position of the cyclohexane (13 and

14) led to a 3-fold decrease in anti-proliferative activity with

Fig. 2. Polar substitutions to mimic phosphate moiety.

Scheme 1. Synthesis of substituted piperidines. Reagents and conditions: a) Bredereck’s reagent, toluene, 100 °C; b) 2-methylresorcinol, AcOH, 130 °C; c) Ac₂O, pyridine,

DCM; d) Pd/C, H₂, RT; e) acyl chloride, Et₃N, THF, RT; f) K₂CO₃, MeOH, RT; g) 1-Boc-3- or -4-hydroxypiperidine, PPh₃, DIAD, DCM, RT; h) TFA, DCM, RT; i) alkyl bromide, DIPEA,

MeCN, MW 80 °C.

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Table 1

Anti-proliferative activity of substituted piperidine analogs.

Compd

R

MCF7 EC₅₀

(l

M)

SKBR3 EC₅₀

(lM)

3

4

H

A

B

C

D

E

1.63 0.35

1.47 0.02

2.66 0.084

13.3 0.62

1.16 0.20

1.19 0.070

1.7 0.15

11a

11b

11c

11d

11e

11f

11 g

12a

12b

12c

12d

12e

12f

12g

Scheme 3. Synthesis of substituted cyclohexylamines. Reagents and conditions: a)

L-selectride, THF, 0 °C; b) NaBH₄; c) 10, PBu₃, TMAD, benzene, 75 °C; d) TFA, DCM,

RT; e) Pt/C, H₂, MeCN, MeOH, RT; f) Ac₂O, Et₃N, DCM, RT; g) BnBr, K₂CO₃, MeCN; h)

benzoyl chloride, Et₃N, THF, RT; i) DIPEA, MeCN, MW 120 °C; j) AcOH, formalde-

hyde, NaBH₃CN, MeOH.

7

2.6

4

1.3

3.13 0.071

4.86 0.032

>50

2.1 0.58

6.27 0.014

>50

F

>50

1.9 0.34

2.1 0.78

1.80 0.064

2.5 0.31

G

A

B

C

D

E

formaldehyde, and sodium cyanoborohydride, dimethylated

amines 20af/bf were produced.

4

2.4

3.0 0.51

3

9

1.6

2.0

As depicted in Table 3, the addition of a single methyl group

onto the exocyclic amine signiﬁcantly decreased potency (18a/b

vs 16a/b). Homologation of the methyl to produce the ethyl deriva-

tive 19aa/ba, led to compounds that also manifested decreased

activity. Interestingly, the addition of a second methyl group onto

18a/b restored potency, but only for the anti analog (20af). This

trend continued with the bulkier amide, benzylamine, and cyclo-

propyl substituents (19ab/ac/ae vs 19bb/bc/be). We therefore

4

1.1

F

G

10.5 0.65

7.35 0.083

20.3

nt

Table 2

Anti-proliferative activity of cyclohexanol and cyclohexylamine analogs.

Scheme 2. Synthesis of cyclohexanol and cyclohexylamine analogs. Reagents and

conditions: a) cyclohexane-1,3- or -1,4-diol, PBu₃, TMAD, benzene, 75 °C; b) i. 3- or

4-N-Boc-aminocyclohexane, PBu₃, TMAD, benzene, 75 °C; ii. TFA, DCM, RT.

Compd

R

MCF7 EC₅₀

(lM)

SKBR3 EC₅₀

(lM)

3

1.6 0.35

1.2 0.20

respect to parent compound 3. A similar decrease was observed for

the compound containing an alcohol at the 4-position (15). In con-

trast, the inclusion of an exocyclic amine at the 4-position

(diastereomers 16a/b) retained activity as compared to 4. We

sought to utilize these diastereomers as a tool and explore both

the size and nature of the surrounding binding pocket.

4

1.47 0.021

4.2 0.59

5.8 0.12

4.7 0.53

1.6 0.22

0.9 0.19

1.19 0.073

6.9 0.43

3.8 0.12

5.4 0.61

0.8 0.36

0.8 0.30

13

The syn- and anti-alcohols 17a/b were synthesized by selective

reduction of tert-butyl methyl(4-oxocyclohexyl)carbamate using

NaBH₄or L-Selectride (Scheme 3) to access the monomethylated

analogs. Mitsunobu conditions were then used to couple the

resulting alcohols with phenol 10, before treatment with acid to

remove the t-butyl carbamate and yield methylamines 18a/b.

The remaining mono- and di-substituted analogs were accessed

by subjecting compounds 16a/b to the conditions depicted in

Scheme 3. For example, the reaction of 16a/b with acetonitrile in

the presence of platinum on carbon and hydrogen gas led to mono

alkylation and furnished ethylamines 19aa/ba. Amides 19ab/bb

were obtained upon the exposure of 16a/b to acetic anhydride.

Nucleophilic substitution of benzyl bromide with 16a/b yielded

benzylamines 19ac/bc. Benzamides 19ad/bd were obtained

after the addition of benzoyl chloride to 16a/b. The mono- and

di-substituted cyclopropyl analogs, 19ae/be and 19ae/be, were

accessed via nucleophilic substitution of the corresponding

mesylate. Finally, after treatment of 16a/b with acetic acid,

14

15

16a

16b

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L.K. Forsberg et al. / Bioorganic & Medicinal Chemistry xxx (2018) xxx–xxx

Table 3

Anti-proliferative activity of 4-cyclohexylamine derivatives.

Compd

R

MCF7 EC₅₀

(l

M)

SKBR3 EC₅₀

(

lM)

Compd

R

MCF7 EC₅₀

(lM)

SKBR3 EC₅₀(lM)

16a

1.6 0.22

0.8 0.36

16b

0.9 0.19

0.8 0.30

18a

6.8 0.77

4.2 0.33

18b

4

1.1

3.4 0.15

19aa

8.9 0.76

7.5 0.45

19ba

7.76 0.08

6.2 0.32

19ab

19ac

19ad

1.16 0.01

1.92 0.08

>50

1.74 0.71

1.6 0.64

>50

19bb

19bc

19bd

>50

19ae

20ae

0.97 0.05

14.5 0.98

1.0 0.14

19be

20be

4.98 0.06

12.3 0.65

3.6 0.21

14.1 0.64

12.8 0.31

20af

1.2 0.23

1.0 0.15

20bf

3.80 0.08

3.49 0.07

sought to combine the beneﬁcial effects of the aliphatic bulk in

19ae with a hydrogen bond acceptor, by the inclusion of cyclic

ethers. Cyclic ethers of various sizes were accessed via nucleophilic

substitution of the corresponding mesylates with amines 16a/b as

described in Scheme 4.

activity, however, enlargement of the ring via tetrahydropyran

analogs 19ai/bi, resulted in compounds that manifested decreased

activity. Mono- and di-substituted compounds manifested similar

anti-proliferative potencies, with exception of the anti tetrahydro-

furan analogs 19ag and 20ag.

Upon their preparation, the ethereal compounds were evalu-

ated against the MCF7 and SKBR3 breast cancer cell lines. As shown

in Table 4, the tetrahydrofuran analogs 19ag/bg maintained the

potency exhibited by the parent compounds, 16a/b. Interestingly,

extension of this moiety via the addition of a methylene group

(19ah/bh), produced compounds with similar anti-proliferative

Neither the piperidine analogs (Table 1) nor the cyclohexy-

lamine analogs (Tables 3 and 4) provided molecules that exhibited

improved anti-proliferative activity versus the respective parent

compounds. In light of this, we replaced the coumarin core with

a phenylcyclohexane moiety, which was recently shown to exhibit

potent C-terminal Hsp90 inhibition.²²The synthesis of this core

began via the diastereoselective reduction of commercially avail-

able ketone 21 with L-Selectride (Scheme 5). Conversion of the

resulting alcohol 22, to the sulfonate ester 23, allowed subsequent

nucleophilic substitution with an azide, which after palladium-cat-

alyzed reduction, yielded the primary amine 24. Upon coupling

with the acid chloride 9 and subsequent solvolysis of the sulfonate

ester, phenol 25 was produced and then subjected to Mitsunobu

etheriﬁcation with the Boc-protected hydroxypiperidines and

cyclohexanediols. Finally, removal of the carbamate and subse-

quent functionalization with alkyl bromides led to the ﬁnal com-

pounds, 26a–h.

Once synthesized, these analogs were evaluated for anti-prolif-

erative activity against the MCF-7 and SBR3 breast cancer cell lines.

As shown in Table 5, replacement of the coumarin core with the

Scheme 4. Synthesis of substituted cyclohexylamines. Reagents and conditions: a)

DIPEA, MeCN, MW 120 °C.

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Table 4

Anti-proliferative activity of ring-constrained ether analogs.

Compd

R

MCF7 EC₅₀

(l

M)

SKBR3 EC₅₀

(

lM)

Compd

R

MCF7 EC₅₀

(l

M)

SKBR3 EC₅₀(lM)

16a

1.6 0.22

0.8 0.36

16b

0.9 0.19

0.8 0.30

19ag

19ah

0.89 0.05

0.81 0.02

19bg

19bh

1.04 0.09

2.2 0.31

1.4 0.17

1.48 0.09

1.3 0.29

1.5 0.43

19ai

4.8 0.39

3.9 0.46

5.8 0.18

19bi

5.6 0.33

2.3 0.22

4.8 0.55

2.5 0.11

20ag

6.31 0.06

20bg

20ah

20ai

2.6 0.28

5.3 0.65

3.0 0.20

4.8 0.38

20bh

20bi

1.6 0.41

7.1 0.66

0.98 0.08

6.6 0.53

phenylcyclohexane ring system led to a 3-fold increase in potency

for all piperidine analogs (26a–f). Conversely, the aminocyclohex-

ane analogs (26g/h) manifested a reduced potency as compared to

the parent compounds. As observed with the coumarin analogs, the

4-substituted piperidine analogs (26e/f) were more potent than

the corresponding 3-substituted piperidine analogs (26b/c). The

data suggest that the phenylcyclohexane core enables the hydro-

gen bond acceptor at the 4 position to project into a more favorable

binding environment.

The syn- and anti-cyclohexylamine derivatives 16a/b and the

anti-dimethylamine 20af were further investigated for their ability

to induce the degradation of Hsp90-dependent substrates. As

shown in the western blot in Fig. 3, these compounds induced

the degradation of Hsp90-dependent client proteins CDK6, ERa,

and HER2, while the levels of Hsp90 and Hsp70 remained constant

conﬁrming that these molecules do not induce the heat shock

response. Degradation of client proteins without induction of the

heat shock response is a hallmark of cytotoxic Hsp90 C-terminal

inhibition. The syn-cyclohexylamine 16b was found to be more

efﬁcacious than anti-cyclohexylamines, 16a and 20af, suggesting

the syn orientation is beneﬁcial for Hsp90 inhibition.

3. Conclusions

A series of compounds was designed and synthesized to project

hydrogen bond donors and acceptors into unexplored regions of

the binding pocket. Although these compounds did not manifest

improved potency, structure-activity relationships suggest the

presence of a large, relatively hydrophobic pocket surrounding

the noviose surrogate. In addition, it was found that replacement

of the coumarin core with a phenylcyclohexane ring system led

to analogs that manifested improved anti-proliferative activity,

which is in agreement with previous reports, suggesting that this

ring system can be utilized to access more efﬁcacious compounds.

Scheme 5. Synthesis of phenylcyclohexane core analogs. Reagents and conditions:

a) L-selectride, THF, 0 °C; b) Et₃N, MsCl, THF, RT; c) NaN₃, DMF, 50 °C; d) Pd/C, H₂,

EtOAc, RT; e) Et₃N, THF, RT; f) NaOH, EtOH, 85 °C; g) PBu₃, TMAD, benzene, 75 °C; h)

TFA, DCM, RT; i) alkyl bromide, DIPEA, MeCN, MW 120 °C.

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Table 5

Anti-proliferative activity of phenylcyclohexane analogs.

Compd

X

R

MCF7 EC₅₀

0.5 0.11

0.625 0.0003

1.0

(

l

M)

SKBR3 EC₅₀(lM)

26a

26b

H

0.42 0.048

0.45 0.017

26c

3

0.9 0.1

26d

26e

H

0.4 0.20

0.38 0.064

0.33 0.01

0.285 0.0096

26f

0.52 0.046

0.363 0.007

26g

26h

C

D

–

2

5

1.0

1.21

1.6 0.32

3.4 0.14

values were adjusted accordingly. IC₅₀values were calculated from

separate experiments performed in triplicate using GraphPad

Prism.

4.2. Western blot

MCF-7 cells were cultured as described above and treated with

various concentrations of drug, GDA in DMSO (1% DMSO ﬁnal con-

centration), or vehicle (DMSO) for 24 h. Cells were harvested in

cold PBS and lysed in RIPA lysis buffer containing 1 mM PMSF,

2 mM sodium orthovanadate, and protease inhibitors on ice for

1 h. Lysates were clariﬁed at 14000 g for 10 min at 4 °C. Protein

concentrations were determined using the Pierce BCA protein

assay kit per the manufacturer’s instructions. Equal amounts of

protein (20 lg) were electrophoresed under reducing conditions,

transferred to a nitrocellulose membrane, and immunoblotted

with the corresponding speciﬁc antibodies. Membranes were incu-

bated with an appropriate horseradish peroxidase-labeled sec-

ondary antibody, developed with a chemiluminescent substrate,

and visualized.

4.3. Synthesis and characterization of ﬁnal compounds

Fig. 3. Cyclohexylamine derivatives induce degradation of Hsp90 client proteins in

MCF7 cells. L represents a concentration equal to 0.5-fold the anti-proliferative

EC₅₀. H represents a concentration equal to 5-fold the anti-proliferative EC_50.

4.3.1. N-(7-((1-(2-Hydroxyethyl)piperidin-3-yl)oxy)-8-methyl-2-oxo-

2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide

(11a)

2-Bromoethanol (1.2 mmol) was added to a solution of DIPEA

(1.5 mmol), 3 (0.05 g, 1.0 mmol), and anhydrous MeCN (2 mL).

The mixture was placed in a sealed microwave safe septum-capped

tube, and heated to 120 °C in a microwave synthesizer. After 1 h

the reaction was complete and the vial was cooled to rt and

quenched with water. The reaction was concentrated and puriﬁed

via column chromatography (SiO₂, 1:10, MeOH: DCM) to afford

11a as a white amorphous solid (30% yield): ¹H NMR (500 MHz,

CDCl₃) d 8.79 (s, 1H), 8.70 (s, 1H), 7.93–7.90 (m, 1H), 7.88 (s, 1H),

7.36 (t, J = 8.0 Hz, 1H), 7.32 (d, J = 8.9 Hz, 1H), 7.12 (d, J = 7.6 Hz,

1H), 7.08 (s, 1H), 7.06 (d, J = 8.7 Hz, 1H), 6.96–6.87 (m, 2H), 4.50–

4.41 (m, 1H), 3.89 (s, 3H), 3.85 (s, 3H), 3.61 (t, J = 5.0 Hz, 2H),

3.02 (d, J = 10.4 Hz, 1H), 2.73 (d, J = 10.3 Hz, 1H), 2.59 (t, J = 4.5

Hz, 2H), 2.41 (t, J = 9.5 Hz, 1H), 2.33–2.26 (m, 5H), 2.09–2.02 (m,

1H), 1.90–1.81 (m, 1H), 1.68–1.54 (m, 2H); ¹³C NMR (126 MHz,

CDCl₃) d 165.62, 159.86, 159.64, 159.43, 157.11, 149.54, 138.72,

131.11, 130.07, 129.29, 128.28, 126.21, 125.72, 124.45, 122.12,

4. Materials and methods

4.1. Anti-proliferation assays

Cells were maintained in a 1:1 mixture of Advanced DMEM/F12

(Gibco) supplemented with non-essential amino acids, l-glutamine

(2 mM), streptomycin (500

10% FBS. Cells were grown to conﬂuence in a humidiﬁed atmo-

sphere (37 °C, 5% CO₂), seeded (2000/well, 100 L) in 96-well

plates, and allowed to attach overnight. Compound or GDA at vary-

ing concentrations in DMSO (1% DMSO ﬁnal concentration) was

added, and cells were returned to the incubator for 72 h. At 72 h,

the number of viable cells was determined using an MTT/PMS cell

proliferation kit (Promega) per the manufacturer’s instructions.

Cells incubated in 1% DMSO were used at 100% proliferation, and

lg/mL), penicillin (100 units/mL), and

l

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L.K. Forsberg et al. / Bioorganic & Medicinal Chemistry xxx (2018) xxx–xxx

7

121.77, 115.36, 115.33, 113.61, 113.27, 111.11, 110.67, 73.72,

4.3.5. Methyl (2-(3-((3-(3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-

carboxamido)-8-methyl-2-oxo-2H-chromen-7-yl)oxy)piperidin-1-yl)

ethyl)carbamate (11e)

59.27, 57.91, 57.63, 56.01, 55.44, 53.16, 29.98, 23.11, 8.45; HRMS

(ESI+) m/z [M+Na] calculated for C₃₂H₃₄N₂O₇Na: 581.2264, found

581.2264.

Compound 11e was obtained following the procedure for the

synthesis of 11a as a white amorphous solid (40%): ¹H NMR

(500 MHz, CDCl₃) d 8.80 (s, 1H), 8.70 (s, 1H), 7.92 (dd, J = 8.6, 2.4

Hz, 1H), 7.89 (d, J = 2.4 Hz, 1H), 7.36 (t, J = 8.0 Hz, 1H), 7.33 (d, J

= 8.7 Hz, 1H), 7.12 (dt, J = 7.6, 1.3 Hz, 1H), 7.09 (dd, J = 2.6, 1.6 Hz,

1H), 7.07 (d, J = 8.6 Hz, 1H), 6.95–6.90 (m, 2H), 5.35 (bs, 1H), 4.51

(bs, 1H), 3.90 (s, 3H), 3.86 (s, 3H), 3.66 (s, 3H), 3.36–3.26 (m, 2H),

3.03 (bs, 1H), 2.76 (bs, 1H), 2.64–2.54 (m, 2H), 2.46–2.39 (m, 1H),

2.35–2.25 (m, 4H), 2.06 (s, 1H), 1.88 (s, 1H), 1.74–1.54 (m, 2H);

¹³C NMR (126 MHz, CDCl₃) d 165.66, 159.88, 159.65, 159.42,

157.25, 156.95, 149.52, 138.71, 131.11, 130.07, 129.30, 128.30,

126.16, 125.78, 124.50, 122.12, 121.78, 115.38, 115.33, 113.67,

113.27, 111.11, 110.72, 73.10, 57.22, 57.05, 56.01, 55.45, 53.23,

52.30, 37.52, 29.77, 22.57, 8.45; HRMS (ESI+) m/z [M+H] calculated

for C₃₃H₃₇N₃O₈S: 635.2301, found 635.2298.

4.3.2. 3⁰,6-Dimethoxy-N-(7-((1-(2-methoxyethyl)piperidin-3-yl)oxy)-

8-methyl-2-oxo-2H-chromen-3-yl)-[1,1⁰-biphenyl]-3-carboxamide

(11b)

Compound 11b was obtained following the procedure for the

synthesis of 11a as a white amorphous solid (33%): ¹H NMR

(500 MHz, CDCl₃) d 8.78 (s, 1H), 8.69 (s, 1H), 7.90 (d, J = 8.6 Hz,

1H), 7.88 (s, 1H), 7.35 (t, J = 7.9 Hz, 1H), 7.31 (d, J = 8.6 Hz, 1H),

7.11 (d, J = 8.4 Hz, 1H), 7.08 (s, 1H), 7.05 (d, J = 8.6 Hz, 1H), 6.95–

6.90 (m, 2H), 4.48 (tt, J = 9.0, 4.0 Hz, 1H), 3.88 (s, 3H), 3.85 (s,

3H), 3.51 (t, J = 5.4 Hz, 2H), 3.34 (s, 3H), 3.19 (d, J = 8.1 Hz, 1H),

2.88 (d, J = 10.9 Hz, 1H), 2.64 (t, J = 5.3 Hz, 2H), 2.30 (s, 3H), 2.22–

2.17 (m, 1H), 2.14–2.07 (m, 2H), 1.85–1.78 (m, 1H), 1.75–1.65

(m, 1H), 1.51–1.41 (m, 1H); 13C NMR (126 MHz, CDCl₃) d 165.55,

159.82, 159.62, 159.39, 157.26, 149.47, 138.69, 131.06, 130.03,

129.25, 128.24, 126.18, 125.66, 124.46, 122.09, 121.68, 115.32,

115.30, 113.51, 113.22, 111.07, 110.90, 74.00, 70.13, 59.03, 58.31,

57.97, 55.97, 55.40, 53.71, 30.34, 23.19, 8.43; HRMS (ESI+) m/z

[M+H] calculated for C₃₃H₃₇N₂O₇: 573.2601, found 573.2595.

4.3.6. Methyl 4-(3-((3-(3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-

carboxamido)-8-methyl-2-oxo-2H-chromen-7-yl)oxy)piperidin-1-yl)

butanoate (11f)

Compound 11f was obtained following the procedure for the

synthesis of 11a as a white amorphous solid (36%): ¹H NMR

(500 MHz, CDCl₃) d 8.80 (s, 1H), 8.70 (s, 1H), 7.92 (dd, J = 8.6, 2.4

Hz, 1H), 7.89 (d, J = 2.4 Hz, 1H), 7.36 (t, J = 7.9 Hz, 1H), 7.33 (d, J

= 8.6 Hz, 1H), 7.12 (ddd, J = 7.6, 1.6, 1.0 Hz, 1H), 7.09 (dd, J = 2.6,

1.5 Hz, 1H), 7.07 (d, J = 8.6 Hz, 1H), 6.96–6.91 (m, 2H), 4.42 (bs,

1H), 3.90 (s, 3H), 3.86 (s, 3H), 3.66 (s, 3H), 3.06 (bs, 1H), 2.75 (bs,

1H), 2.41 (bs, 2H), 2.34 (t, J = 7.1 Hz, 2H), 2.31 (s, 3H), 2.22–2.16

(m, 2H), 2.13–2.05 (m, 2H), 1.86–1.77 (m, 3H), 1.54–1.44 (m,

1H); ¹³C NMR (126 MHz, CDCl₃) d 174.16, 165.63, 159.87, 159.71,

159.44, 157.35, 149.54, 138.74, 131.13, 130.08, 129.31, 128.30,

126.24, 125.73, 124.56, 122.14, 121.72, 115.40, 115.34, 113.54,

113.29, 111.12, 110.90, 74.19, 57.87, 57.66, 56.02, 55.46, 53.35,

51.71, 31.99, 30.42, 23.32, 22.21, 8.47; HRMS (ESI+) m/z [M+Na]

calculated for C₃₅H₃₈N₂O₈Na: 615.2706, found 615.2687.

4.3.3. N-(7-((1-(3-Hydroxypropyl)piperidin-3-yl)oxy)-8-methyl-2-

oxo-2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-

carboxamide (11c)

Compound 11c was obtained following the procedure for the

synthesis of 11a as a white amorphous solid (35%): ¹H NMR

(500 MHz, CDCl₃) d 8.79 (s, 1H), 8.70 (s, 1H), 7.91 (dd, J = 8.5, 2.4

Hz, 1H), 7.89 (d, J = 2.4 Hz, 1H), 7.36 (t, J = 7.9 Hz, 1H), 7.33 (d, J

= 8.6 Hz, 1H), 7.12 (ddd, J = 7.9, 1.6, 1.0 Hz, 1H), 7.09 (dd, J = 2.6,

1.6 Hz, 1H), 7.06 (d, J = 8.6 Hz, 1H), 6.93 (ddd, J = 8.5, 2.7, 1.0 Hz,

1H), 6.90 (d, J = 8.8 Hz, 1H), 4.46 (dt, J = 8.3, 3.9 Hz, 1H), 3.89 (s,

3H), 3.86 (s, 3H), 3.84–3.78 (m, 2H), 3.07–2.97 (bs, 1H), 2.76–

2.69 (bs, 1H), 2.66 (t, J = 5.7 Hz, 2H), 2.51–2.39 (bs, 0H), 2.38–

2.26 (m, 5H), 2.04–1.94 (m, 1H), 1.91–1.83 (m, 1H), 1.77–1.69

(m, 2H), 1.66–1.55 (m, 2H); ¹³C NMR (126 MHz, CDCl₃) d 165.60,

159.85, 159.65, 159.43, 157.03, 149.53, 138.72, 131.10, 130.07,

129.29, 128.27, 126.22, 125.72, 124.47, 122.12, 121.75, 115.39,

115.32, 113.59, 113.27, 111.10, 110.48, 73.28, 64.82, 59.26, 57.98,

56.00, 55.44, 53.77, 29.62, 27.31, 22.77, 8.41; HRMS (ESI+) m/z

[M+H] calculated for C₃₃H₃₇N₂O₇: 573.2601, found 573.2600.

4.3.7. 3⁰,6-Dimethoxy-N-(8-methyl-7-((1-(2-(methylsulfonamido)

ethyl)piperidin-3-yl)oxy)-2-oxo-2H-chromen-3-yl)-[1,1⁰-biphenyl]-3-

carboxamide (11g)

Compound 11gwas obtained following the procedure for the

synthesis of 11a as a white amorphous solid (32%): ¹H NMR

(500 MHz, CDCl₃) d 8.78 (s, 1H), 8.69 (s, 1H), 7.91 (dd, J = 8.7, 2.5

Hz, 1H), 7.88 (d, J = 2.3 Hz, 1H), 7.38–7.31 (m, 2H), 7.12 (dt, J =

7.6, 1.2 Hz, 1H), 7.09 (dd, J = 2.6, 1.6 Hz, 1H), 7.06 (d, J = 8.6 Hz,

1H), 6.95–6.88 (m, 2H), 5.11 (bs, 1H), 4.44 (tt, J = 7.9, 3.8 Hz, 1H),

3.89 (s, 3H), 3.85 (s, 3H), 3.23–3.18 (m, 2H), 3.03–2.96 (m, 1H),

2.93 (s, 3H), 2.75–2.67 (m, 1H), 2.61 (t, J = 5.8 Hz, 2H), 2.41–2.33

(m, 1H), 2.31 (d, J = 1.6 Hz, 3H), 2.29–2.21 (m, 1H), 2.09–2.01 (m,

1H), 1.90–1.82 (m, 1H), 1.67–1.51 (m, 2H); ¹³C NMR (126 MHz,

CDCl₃) d 165.58, 159.83, 159.56, 159.39, 156.92, 149.49, 138.69,

131.05, 130.03, 129.27, 128.27, 126.13, 125.74, 124.38, 122.09,

121.78, 115.31, 115.28, 113.65, 113.21, 111.08, 110.65, 73.36,

57.32, 56.73, 55.98, 55.42, 53.11, 40.16, 39.69, 29.85, 22.83, 8.46;

4.3.4. 3⁰,6-Dimethoxy-N-(7-((1-(3-methoxypropyl)piperidin-3-yl)

oxy)-8-methyl-2-oxo-2H-chromen-3-yl)-[1,1⁰-biphenyl]-3-

carboxamide (11d)

Compound 11d was obtained following the procedure for the

synthesis of 11a as a white amorphous solid (25%): ¹H NMR

(500 MHz, CDCl₃) d 8.79 (s, 1H), 8.70 (s, 1H), 7.91 (dd, J = 8.5, 2.4

Hz, 1H), 7.89 (d, J = 2.4 Hz, 1H), 7.36 (t, J = 7.9 Hz, 1H), 7.31 (d, J

= 8.6 Hz, 1H), 7.12 (dt, J = 7.6, 1.2 Hz, 1H), 7.09 (dd, J = 2.7, 1.6 Hz,

1H), 7.06 (d, J = 8.5 Hz, 1H), 6.94–6.89 (m, 2H), 4.42 (tt, J = 8.7,

4.0 Hz, 1H), 3.89 (s, 3H), 3.85 (s, 3H), 3.40 (t, J = 6.4 Hz, 2H), 3.31

(s, 3H), 3.10–3.04 (m, 1H), 2.78–2.72 (m, 1H), 2.46 (t, J = 7.6 Hz,

2H), 2.31 (s, 3H), 2.16 (t, J = 9.8 Hz, 1H), 2.13–2.04 (m, 2H), 1.86–

1.79 (m, 1H), 1.79–1.72 (m, 2H), 1.68–1.58 (m, 1H), 1.53–1.43

(m, 1H); ¹³C NMR (126 MHz, CDCl₃) d 165.55, 159.82, 159.63,

159.40, 157.35, 149.49, 138.70, 131.07, 130.04, 129.26, 128.24,

126.20, 125.64, 124.45, 122.10, 121.68, 115.39, 115.31, 113.49,

113.23, 111.07, 110.90, 74.32, 71.15, 58.73, 58.04, 55.97, 55.57,

55.41, 53.44, 30.46, 27.20, 23.37, 8.45; HRMS (ESI+) m/z [M+H] cal-

culated for C₃₄H₃₈N₂O₇: 586.2679, found 586.2681.

HRMS (ESI+) m/z [M+Na] calculated for

C₃₃H₃₇N₃O₈SNa:

658.2199, found 658.2205.

4.3.8. N-(7-((1-(2-hydroxyethyl)piperidin-4-yl)oxy)-8-methyl-2-oxo-

2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide

(12a)

2-Bromoethanol (1.2 mmol) was added to a solution of DIPEA

(1.5 mmol), 4 (0.05 g, 1.0 mmol), and anhydrous MeCN (2 mL).

The mixture was placed in a sealed microwave safe septum-capped

tube, and heated to 120 °C in a microwave synthesizer. After 1 h

the reaction was complete and the vial was cooled to rt and

Please cite this article in press as: Forsberg L.K., et al. Bioorg. Med. Chem. (2018), https://doi.org/10.1016/j.bmc.2018.04.028

8

L.K. Forsberg et al. / Bioorganic & Medicinal Chemistry xxx (2018) xxx–xxx

quenched with water. The reaction was concentrated and puriﬁed

via column chromatography (SiO₂, 1:10, MeOH: DCM) to afford

12a as a white amorphous solid (34% yield): ¹H NMR (500 MHz,

CDCl3) d 8.79 (s, 1H), 8.70 (s, 1H), 7.91 (dd, J = 8.5, 2.4 Hz, 1H),

7.88 (d, J = 2.4 Hz, 1H), 7.36 (t, J = 7.9 Hz, 1H), 7.32 (d, J = 8.6 Hz,

1H), 7.12 (dt, J = 7.6, 1.2 Hz, 1H), 7.09 (dd, J = 2.6, 1.5 Hz, 1H),

7.06 (d, J = 8.7 Hz, 1H), 6.93 (ddd, J = 8.3, 2.6, 1.0 Hz, 1H), 6.86 (d,

J = 8.7 Hz, 1H), 4.51 (bs, 1H), 3.89 (s, 3H), 3.85 (s, 3H), 3.66 (t, J =

5.3 Hz, 2H), 3.26 (bs, 1H), 2.79 (t, J = 9.1 Hz, 2H), 2.62 (t, J = 5.4

Hz, 2H), 2.55 (bs, 2H), 2.34 (s, 3H), 2.11–2.00 (m, 2H), 1.97–1.88

(m, 2H); ¹³C NMR (126 MHz, CDCl₃) d 165.60, 159.86, 159.60,

159.42, 156.87, 149.53, 138.70, 131.10, 130.04, 129.28, 128.26,

126.16, 125.67, 124.41, 122.10, 121.74, 115.34, 115.29, 113.52,

113.23, 111.10, 110.54, 72.54, 59.50, 57.91, 56.00, 55.43, 50.10

(bs, 2H), 2.37 (bs, 2H), 2.34 (s, 3H), 2.12–1.96 (m, 4H); ¹³C NMR

(126 MHz, CDCl₃) 165.64, 159.91, 159.53, 159.43, 156.28,

149.52, 138.69, 131.13, 130.05, 129.31, 128.31, 126.11, 125.88,

124.24, 122.11, 121.98, 115.36, 115.13, 113.86, 113.25, 111.13,

110.32, 70.33 (2C), 58.88, 56.02, 55.88, 55.45, 49.42 (2C), 28.98

(2C), 25.87, 8.58; HRMS (ESI+) m/z [M+H] calculated for

d

C₃₄H₃₈N₂O₇: 586.2679, found 586.2675.

4.3.12. Methyl (2-(4-((3-(3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-

carboxamido)-8-methyl-2-oxo-2H-chromen-7-yl)oxy)piperidin-1-yl)

ethyl)carbamate (12e)

Compound 12e was obtained following the procedure for the

synthesis of 12a as a white amorphous solid (31%): ¹H NMR

(500 MHz, CDCl₃) d 8.79 (s, 1H), 8.70 (s, 1H), 7.91 (dd, J = 8.5, 2.4

Hz, 1H), 7.89 (d, J = 2.4 Hz, 1H), 7.36 (dd, J = 8.3, 7.6 Hz, 1H), 7.32

(d, J = 8.2 Hz, 1H), 7.12 (ddd, J = 7.6, 1.6, 1.0 Hz, 1H), 7.09 (dd, J =

2.6, 1.6 Hz, 1H), 7.07 (d, J = 8.6 Hz, 1H), 6.93 (ddd, J = 8.3, 2.6, 1.0

Hz, 1H), 6.87 (d, J = 8.7 Hz, 1H), 5.22 (bs, 1H), 4.47 (bs, 1H), 3.90

(s, 3H), 3.86 (s, 3H), 3.68 (s, 3H), 3.33–3.22 (m, 2H), 2.69 (bs, 2H),

2.50 (t, J = 6.0 Hz, 2H), 2.39 (bs, 3H), 2.34 (s, 3H), 2.06–1.94 (m,

2H), 1.92–1.80 (m, 2H); ¹³C NMR (126 MHz, CDCl₃) d 165.63,

159.87, 159.66, 159.43, 157.17, 157.02, 149.56, 138.72, 131.12,

130.07, 129.30, 128.28, 126.20, 125.65, 124.47, 122.12, 121.72,

115.37, 115.35, 113.47, 113.26, 111.10, 110.62, 73.07, 57.10,

56.02, 55.45, 52.20, 50.22 (2C), 37.92, 30.91 (2C), 8.54; HRMS

(ESI+) m/z [M+H] calculated for C₃₃H₃₇N₃O₈S: 635.2301, found

635.2296.

(2C), 30.67 (2C), 8.52; HRMS (ESI+) m/z [M+Na] calculated for C₃₂

₃₄N₂O₇Na: 581.2264, found 581.2272.

-

H

4.3.9. 3⁰,6-Dimethoxy-N-(7-((1-(2-methoxyethyl)piperidin-4-yl)oxy)-

8-methyl-2-oxo-2H-chromen-3-yl)-[1,1⁰-biphenyl]-3-carboxamide

(12b)

Compound 12b was obtained following the procedure for the

synthesis of 12a as a white amorphous solid (37%): ¹H NMR

(500 MHz, CDCl₃) d 8.79 (s, 1H), 8.69 (s, 1H), 7.91 (dd, J = 8.6, 2.4

Hz, 1H), 7.88 (d, J = 2.4 Hz, 1H), 7.39–7.33 (m, 2H), 7.12 (dt, J =

7.6, 1.3 Hz, 1H), 7.10–7.05 (m, 2H), 6.93 (ddd, J = 8.3, 2.6, 0.9 Hz,

1H), 6.85 (d, J = 8.7 Hz, 1H), 4.69 (bs, 1H), 3.90 (s, 3H), 3.85 (s,

3H), 3.84–3.77 (m, 2H), 3.37 (s, 3H), 3.19–3.08 (m, 4H), 3.04 (bs,

2H), 2.49 (bs, 2H), 2.35 (s, 3H), 2.17–2.08 (m, 2H); ¹³C NMR (126

MHz, CDCl₃) d 165.64, 159.92, 159.48, 159.43, 155.98, 149.50,

138.68, 131.13, 130.03, 129.31, 128.31, 126.08, 125.95, 124.16,

122.10, 122.06, 115.37, 115.07, 113.99, 113.23, 111.13, 110.19,

69.27, 68.15, 59.14, 57.53, 56.02, 55.45, 49.55 (2C), 28.27 (2C),

4.3.13. Methyl 4-(4-((3-(3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-

carboxamido)-8-methyl-2-oxo-2H-chromen-7-yl)oxy)piperidin-1-yl)

butanoate (12f)

Compound 12f was obtained following the procedure for the

synthesis of 12a as a white amorphous solid (38%): ¹H NMR

(500 MHz, CDCl₃) d 8.79 (s, 1H), 8.70 (s, 1H), 7.91 (dd, J = 8.5, 2.4

Hz, 1H), 7.89 (d, J = 2.4 Hz, 1H), 7.39–7.34 (m, 1H), 7.32 (d, J =

8.6 Hz, 1H), 7.12 (ddd, J = 7.6, 1.7, 1.0 Hz, 1H), 7.09 (dd, J = 2.6,

1.5 Hz, 1H), 7.07 (d, J = 8.6 Hz, 1H), 6.93 (ddd, J = 8.3, 2.7, 1.0 Hz,

1H), 6.87 (d, J = 8.6 Hz, 1H), 4.46 (bs, 1H), 3.90 (s, 3H), 3.86 (s,

3H), 3.68 (s, 3H), 2.70 (bs, 2H), 2.44–2.34 (m, 6H), 2.33 (s, 3H),

2.06–1.97 (m, 2H), 1.92–1.80 (m, 4H); ¹³C NMR (126 MHz, CDCl₃)

d 174.12, 165.62, 159.86, 159.67, 159.43, 157.06, 149.56, 138.72,

131.11, 130.07, 129.30, 128.28, 126.21, 125.65, 124.50, 122.12,

121.69, 115.36, 115.34, 113.44, 113.26, 111.10, 110.65, 73.14,

57.74, 56.01, 55.45, 51.72, 50.32 (2C), 32.11, 30.84 (2C), 22.41,

8.53; HRMS (ESI+) m/z [M+Na] calculated for C₃₅H₃₈N₂O₈Na:

615.2706, found 615.2699.

8.53; HRMS (ESI+) m/z [M+H] calculated for

573.2601, found 573.2609.

C₃₃H₃₇N₂O₇:

4.3.10. N-(7-((1-(3-Hydroxypropyl)piperidin-4-yl)oxy)-8-methyl-2-

oxo-2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-

carboxamide (12c)

Compound 12c was obtained following the procedure for the

synthesis of 12a as a white amorphous solid (39%): ¹H NMR

(500 MHz, CDCl₃) d 8.79 (s, 1H), 8.70 (s, 1H), 7.91 (dd, J = 8.6, 2.4

Hz, 1H), 7.89 (d, J = 2.4 Hz, 1H), 7.36 (t, J = 8.4, 7.6 Hz, 1H), 7.32

(d, J = 8.6 Hz, 1H), 7.12 (ddd, J = 7.6, 1.6, 1.0 Hz, 1H), 7.09 (dd, J =

2.6, 1.6 Hz, 1H), 7.07 (d, J = 8.6 Hz, 1H), 6.93 (ddd, J = 8.3, 2.6, 1.0

Hz, 1H), 6.86 (d, J = 8.7 Hz, 1H), 4.51 (s, 1H), 3.90 (s, 3H), 3.86 (s,

3H), 3.82 (t, J = 5.2 Hz, 2H), 2.75 (bs, 2H), 2.68 (t, J = 5.6 Hz, 2H),

2.55 (bs, 2H), 2.33 (s, 3H), 2.06–1.97 (m, 2H), 1.96–1.85 (m, 2H),

1.79–1.72 (m, 2H); ¹³C NMR (126 MHz, CDCl₃) d 165.63, 159.88,

159.63, 159.44, 156.86, 149.56, 138.72, 131.13, 130.07, 129.30,

128.28, 126.20, 125.69, 124.42, 122.13, 121.77, 115.35, 115.34,

113.55, 113.26, 111.12, 110.53, 72.22, 64.65, 59.07, 56.02, 55.45,

50.38 (2C), 30.69 (2C), 27.34, 8.54; HRMS (ESI+) m/z [M+H] calcu-

lated for C₃₃H₃₇N₂O₇: 573.2601, found 573.2607.

4.3.14. 3⁰,6-Dimethoxy-N-(8-methyl-7-((1-(2-(methylsulfonamido)

ethyl)piperidin-4-yl)oxy)-2-oxo-2H-chromen-3-yl)-[1,1⁰-biphenyl]-3-

carboxamide (12g)

Compound 12 g was obtained following the procedure for the

synthesis of 12a as a white amorphous solid (41%): ¹H NMR

(500 MHz, CDCl₃) d 8.80 (s, 1H), 8.70 (s, 1H), 7.92 (dd, J = 8.6, 2.4

Hz, 1H), 7.89 (d, J = 2.4 Hz, 1H), 7.40–7.31 (m, 2H), 7.12 (dt, J =

7.8, 1.2 Hz, 1H), 7.09 (dd, J = 2.6, 1.7 Hz, 1H), 7.07 (d, J = 8.6 Hz,

1H), 6.93 (ddd, J = 8.2, 2.7, 0.9 Hz, 1H), 6.87 (d, J = 8.6 Hz, 1H),

5.10 (bs, 1H), 4.51 (bs, 1H), 3.90 (s, 3H), 3.86 (s, 3H), 3.23 (t, J =

5.6 Hz, 2H), 2.99 (s, 3H), 2.71 (bs, 2H), 2.61 (t, J = 6.3, 5.2 Hz, 2H),

2.47 (bs, 2H), 2.34 (s, 3H), 2.06–1.97 (m, 2H), 1.95–1.83 (m, 2H);

¹³C NMR (126 MHz, CDCl₃) d 165.65, 159.90, 159.64, 159.44,

156.86, 149.57, 138.72, 131.14, 130.07, 129.32, 128.30, 126.19,

125.70, 124.43, 122.13, 121.79, 115.36 (2C), 113.57, 113.27,

111.12, 110.55, 72.47, 56.88, 56.03, 55.47, 49.99 (2C), 40.33,

4.3.11. 3⁰,6-Dimethoxy-N-(7-((1-(3-methoxypropyl)piperidin-4-yl)

oxy)-8-methyl-2-oxo-2H-chromen-3-yl)-[1,1⁰-biphenyl]-3-

carboxamide (12d)

Compound 12d was obtained following the procedure for the

synthesis of 12a as a white amorphous solid (45%): ¹H NMR

(500 MHz, CDCl₃) d 8.80 (s, 1H), 8.70 (s, 1H), 7.91 (dt, J = 8.7, 1.7

Hz, 1H), 7.88 (d, J = 2.2 Hz, 1H), 7.39–7.31 (m, 2H), 7.14–7.10 (m,

1H), 7.10–7.07 (m, 1H), 7.06 (s, 1H), 6.93 (ddd, J = 8.2, 2.6, 1.0 Hz,

1H), 6.85 (d, J = 8.6 Hz, 1H), 4.63 (bs, 1H), 3.90 (s, 3H), 3.86 (s,

3H), 3.46 (t, J = 5.9 Hz, 2H), 3.33 (s, 3H), 3.01–2.85 (m, 4H), 2.79

Please cite this article in press as: Forsberg L.K., et al. Bioorg. Med. Chem. (2018), https://doi.org/10.1016/j.bmc.2018.04.028

L.K. Forsberg et al. / Bioorganic & Medicinal Chemistry xxx (2018) xxx–xxx

9

39.81, 30.70 (2C), 8.56; HRMS (ESI+) m/z [M+Na] calculated for C₃₃

₃₇N₃O₈SNa: 658.2199, found 658.2176.

-

4.3.18. N-(7-(((1r,4r)-4-Aminocyclohexyl)oxy)-8-methyl-2-oxo-2H-

chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide(16a)

H

Compound 16a was obtained following the procedure for the

synthesis of 14 as a white amorphous solid (70% yield): ¹H NMR

(500 MHz, Chloroform-d) d 8.77 (d, J = 4.0 Hz, 1H), 8.71 (d, J = 6.8

Hz, 1H), 7.90–7.82 (m, 2H), 7.38–7.30 (m, 1H), 7.10–7.02 (m, 3H),

6.90 (dt, J = 8.1, 4.6 Hz, 1H), 6.84 (d, J = 8.5 Hz, 1H), 4.53 (d, J =

18.5 Hz, 1H), 3.88 (d, 6H), 3.60–3.54 (m, 1H), 2.24 (s, 3H), 2.09

(m, 2H), 1.88–1.81 (m, 2H), 1.76–1.54 (m, 4H). ¹³C NMR (126

MHz, CDCl₃) d 165.99, 160.44, 159.91, 159.58, 157.02, 155.06,

149.00, 148.83, 138.16, 131.28, 130.53, 129.86, 128.52, 126.22,

126.01, 124.40, 122.88, 121.83, 115.63, 113.52, 111.57, 110.50,

79.00, 77.36, 71.05, 48.29, 30.39, 28.12, 8.17.HRMS (ESI+), m/z [M

+H⁺] calculated for C₃₁H₃₂N₂O₆529.2339; found 529.2333.

4.3.15. N-(7-((3-Hydroxycyclohexyl)oxy)-8-methyl-2-oxo-2H-

chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide (13)

Phenol 9 (0.12 g, 0.298 mmol), and cyclohexane-1,3-diol (0.128

g, 0.597 mmol) were dissolved in anhydrous benzene (2mL). Tri-

butylphosphine (0.147 mL, 0.597 mmol) was added dropwise and

the reaction was stirred at 0 °C. After 5 min the mixture was cov-

ered and diamide (0.103 g, 0.597 mmol) was added. The mixture

was stirred for 15 min and then heated to reﬂux. The reaction

was stirred at reﬂux for 12 h, cooled to rt, concentrated, and puri-

ﬁed via column chromatography (SiO₂, 1:4, EtOAc: hexane) to

afford 12 as a white amorphous solid (35% yield): ¹H NMR (500

MHz, Chloroform-d) d 8.80 (s, 1H), 8.70 (d, J = 2.8 Hz, 1H), 7.93–

7.88 (m, 3H), 7.35 (dt, J = 15.6, 8.3 Hz, 3H), 7.12 (dt, J = 7.6, 1.2

Hz, 2H), 7.10–7.04 (m, 4H), 6.95–6.87 (m, 3H), 4.81 (dt, J = 6.0,

3.2 Hz, 1H), 4.39 (tt, J = 8.9, 3.8 Hz, 1H), 4.14 (ddd, J = 14.5, 9.9,

5.8 Hz, 1H), 3.90 (s, 4H), 3.86 (s, 4H), 3.83–3.77 (m, 1H), 2.33 (s,

3H), 2.13 (dt, J = 12.6, 4.6 Hz, 1H), 2.04–1.98 (m, 1H), 1.94–1.86

(m, 2H), 1.70 (ddt, J = 21.7, 12.5, 7.0 Hz, 3H), 1.60–1.33 (m, 2H).

¹³C NMR (126 MHz, CDCl₃) d 165.67, 159.92, 159.64, 159.47,

157.23, 149.56, 138.75, 131.19, 130.10, 129.31, 128.29, 125.67,

124.43, 122.15, 121.85, 115.39, 113.30, 111.15, 110.90, 75.58,

73.70, 68.47, 67.04, 40.49, 39.11, 34.41, 30.89, 19.27, 8.58. HRMS

(ESI+), m/z [M+H⁺] calculated for C₃₁H₃₁NO_7:530.2179; found

530.2162.

4.3.19. N-(7-(((1s,4s)-4-Aminocyclohexyl)oxy)-8-methyl-2-oxo-2H-

chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide (16b)

Compound 16b was obtained following the procedure for the

synthesis of 14 as a white amorphous solid 70%): ¹H NMR (500

MHz, Methanol d₄) d 8.47 (d, J = 1.3 Hz, 1H), 7.61 (dt, J = 8.6, 1.9

Hz, 1H), 7.57 (d, J = 2.4 Hz, 1H), 7.08–7.03 (m, 2H), 6.82 (dt, J =

7.6, 1.2 Hz, 1H), 6.80–6.76 (m, 2H), 6.63 (ddd, J = 8.2, 2.7, 1.0 Hz,

1H), 6.57 (d, J = 8.7 Hz, 1H), 4.39 (t, J = 3.1 Hz, 1H), 3.60 (d, J = 1.3

Hz, 3H), 3.55 (d, J = 1.3 Hz, 3H), 2.84 (ddt, J = 11.2, 7.9, 3.9 Hz,

1H), 2.06 (s, 3H), 1.92–1.83 (m, 2H), 1.65–1.58 (m, 2H), 1.55–

1.45 (m, 2H), 1.39 (tt, J = 13.6, 3.3 Hz, 2H). ¹³C NMR (126 MHz,

MeOD) d 165.81, 159.77, 159.54, 159.17, 156.47, 149.36, 138.51,

130.90, 129.84, 129.11, 128.16, 125.70, 124.81, 121.93, 121.33,

115.14, 113.01, 111.00, 109.98, 70.03, 55.78, 55.22, 27.85, 25.39,

7.93. HRMS (ESI+), m/z [M+H⁺] calculated for C₃₁H₃₂N₂O₆

529.2339; found 529.2336.

4.3.16. N-(7-((3-Aminocyclohexyl)oxy)-8-methyl-2-oxo-2H-chromen-

3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide (14)

Compound 14 was obtained following the procedure for the

synthesis of 13 as a white amorphous solid (35%). The boc- pro-

tected amine was dissolved in anhydrous dichloromethane

(1mL). Triﬂuoroacetic acid (0.3 mL, 30% vol.) was added to the mix-

ture at 0 °C. The mixture was warmed to rt and after 12 h the reac-

tion was concentrated and puriﬁed via column chromatography

(SiO₂, 1:10, MeOH: DCM) to afford 14 in (70%) yield as a white

solid: ¹H NMR (500 MHz, Methanol d₄) d 8.73 (s, 1H), 7.87 (dd, J

= 8.6, 2.4 Hz, 1H), 7.84 (d, J = 2.4 Hz, 1H), 7.31 (d, J = 8.2 Hz, 2H),

7.10–7.01 (m, 4H), 6.89 (ddd, J = 8.3, 2.6, 0.9 Hz, 1H), 6.83 (d, J =

8.7 Hz, 1H), 4.82 (t, J = 3.1 Hz, 1H), 3.85 (s, 3H), 3.81 (s, 3H),

3.50–3.43 (m, 1H), 2.37–2.32 (m, 2H), 2.28 (s, 3H), 2.07 (d, J =

12.6 Hz, 2H), 2.03–1.96 (m, 2H), 1.76–1.64 (m, 2H). ¹³C NMR

4.3.20. 3⁰,6-Dimethoxy-N-(8-methyl-7-(((1r,4r)-4-

(methylamino)cyclohexyl)oxy)-2-oxo-2H-chromen-3-yl)-[1,1⁰-

biphenyl]-3-carboxamide (18a)

Compound 18a was obtained following the procedure for the

synthesis of 16a as a white amorphous solid 70%): ¹H NMR (500

MHz, Methanol d₄) d 8.02 (s, 1H), 7.18 (dd, J = 8.6, 2.5 Hz, 1H),

7.14 (d, J = 2.4 Hz, 1H), 6.67–6.58 (m, 3H), 6.41–6.34 (m, 4H),

6.22–6.13 (m, 2H), 3.99 (t, J = 2.9 Hz, 1H), 3.17 (s, 3H), 3.12 (s,

3H), 2.62 (p, J = 1.6 Hz, 1H), 2.34 (tt, J = 11.7, 4.1 Hz, 1H), 1.94 (d,

J = 2.6 Hz, 3H), 1.62 (s, 3H), 1.50 (dt, J = 16.0, 2.9 Hz, 2H), 1.26

(dd, J = 13.0, 3.8 Hz, 2H), 1.18–1.07 (m, 2H), 1.04–0.92 (m, 2H).

¹³C NMR (126 MHz, MeOD) d 165.63, 159.55, 159.25, 158.92,

156.15, 149.11, 138.27, 130.65, 129.56, 128.81, 127.88, 125.47,

124.67, 121.65, 121.07, 114.89, 112.71, 110.79, 109.74, 69.73,

56.58, 55.46, 54.90, 29.22, 27.49, 22.88, 7.57. HRMS (ESI+), m/z

[M+H⁺] calculated for C₃₂H₃₄N₂O₆543.2495; found 543.2462.

(126 MHz, MeOD)

d 166.59, 160.67, 160.10, 157.13, 150.21,

139.40, 131.83, 130.74, 129.98, 129.01, 126.59, 125.44, 122.82,

122.37, 116.05, 114.37, 113.93, 111.89, 110.92, 72.74, 56.66,

56.11, 49.98, 47.22, 30.49, 28.76, 19.65, 8.95. HRMS (ESI+), m/z

[M+H⁺] calculated for C₃₁H₃₂N₂O₆529.2339; found 529.2326.

4.3.17. N-(7-((4-Hydroxycyclohexyl)oxy)-8-methyl-2-oxo-2H-

chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide (15)

Compound 15 was obtained following the procedure for the

synthesis of 13 as a white amorphous solid (35%): ¹H NMR (500

MHz, Chloroform-d) d 8.80 (d, J = 1.0 Hz, 2H), 8.70 (d, J = 2.6 Hz,

2H), 7.95–7.87 (m, 5H), 7.39–7.31 (m, 4H), 7.12 (dt, J = 7.6, 1.2

Hz, 2H), 7.10–7.05 (m, 5H), 6.95–6.88 (m, 4H), 4.53 (dt, J = 5.4,

2.7 Hz, 0H), 4.40 (dt, J = 8.6, 4.6 Hz, 1H), 3.90 (s, 6H), 3.86 (s, 7H),

2.36 (d, J = 2.7 Hz, 3H), 2.32 (s, 3H), 2.14 (ddd, J = 12.5, 5.0, 2.5

Hz, 2H), 2.05 (td, J = 8.5, 7.6, 4.3 Hz, 2H), 1.78 (t, J = 4.0 Hz, 1H),

1.64 (ddd, J = 12.8, 8.6, 3.2 Hz, 2H), 1.49 (dq, J = 8.6, 2.8, 2.3 Hz,

2H). ¹³C NMR (126 MHz, CDCl₃) d 165.51, 159.76, 159.56, 159.33,

157.30, 138.61, 131.04, 129.95, 129.17, 128.15, 126.13, 125.53,

124.41, 122.01, 115.24, 113.16, 111.01, 110.66, 75.24, 68.59,

31.54, 28.17, 28.15, 8.32, 1.03. HRMS (ESI+), m/z [M+Na⁺] calcu-

lated for C₃₁H₃₁NO₇552.1998; found 552.2015.

4.3.21. 3⁰,6-Dimethoxy-N-(8-methyl-7-(((1s,4s)-4-

(methylamino)cyclohexyl)oxy)-2-oxo-2H-chromen-3-yl)-[1,1⁰-

biphenyl]-3-carboxamide (18b)

Compound 18b was obtained following the procedure for the

synthesis of 16a as a white amorphous solid 70%): ¹H NMR (500

MHz, Chloroform-d) d 9.39 (d, J = 9.9 Hz, 2H), 9.22 (s, 1H), 8.77

(d, J = 10.0 Hz, 1H), 8.67 (s, 1H), 7.92–7.85 (m, 2H), 7.33 (dt, J =

21.9, 8.1 Hz, 2H), 7.14–7.03 (m, 3H), 6.92 (dd, J = 7.9, 2.5 Hz, 1H),

6.83 (dd, J = 12.0, 8.7 Hz, 1H), 4.68 (d, J = 3.7 Hz, 1H), 4.30 (d, J =

9.6 Hz, 1H), 3.89 (d, J = 6.3 Hz, 3H), 3.85 (s, 3H), 3.06–2.93 (m,

2H), 2.67 (d, J = 5.1 Hz, 5H), 2.32 (s, 2H), 2.29 (s, 1H), 2.22 (d, J =

15.8 Hz, 3H), 2.04–1.92 (m, 5H), 1.70–1.52 (m, 3H), 1.29 (d, J =

33.7 Hz, 3H). ¹³C NMR (126 MHz, CDCl₃)

d 165.34, 159.65,

159.21, 149.32, 138.49, 130.91, 129.86, 129.05, 128.00, 126.94,

125.91, 125.42, 124.07, 122.21, 121.89, 115.13, 113.03, 110.89,

109.74, 55.75, 55.21, 30.28, 29.60, 29.07, 27.85, 26.00, 23.10,

Please cite this article in press as: Forsberg L.K., et al. Bioorg. Med. Chem. (2018), https://doi.org/10.1016/j.bmc.2018.04.028

10

L.K. Forsberg et al. / Bioorganic & Medicinal Chemistry xxx (2018) xxx–xxx

8.22, 7.93. HRMS (ESI+), m/z [M+H⁺] calculated for C₃₂H₃₄N₂O₆

543.2495; found 543.2490.

157.10, 149.53, 138.73, 131.12, 130.07, 129.31, 129.11, 128.31,

126.22, 125.69, 124.43, 122.14, 121.81, 115.34, 113.64, 113.28,

111.11, 110.85, 75.48, 54.42, 49.13, 29.76, 27.59, 8.48. HRMS (ESI

+), m/z [M+H⁺] calculated for C₃₈H₃₈N₂O₆619.2808; found

619.2791.

4.3.22. N-(7-(((1r,4r)-4-(Ethylamino)cyclohexyl)oxy)-8-methyl-2-

oxo-2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-

carboxamide (19aa)

Compound 16a (0.04 g, 0.06 mmol) was dissolved in methanol

(0.6 mL). 10% Pt/C (0.002, 0.008 mmol) and acetonitrile (0.01 mL)

were added to the mixture and the suspension was stirred under

a hydrogen atmosphere. After 25 h, the reaction mixture was ﬁl-

tered through a pad of Celite; the residue was washed with ethyl

acetate. The ﬁltrate was concentrated and puriﬁed by column chro-

matography (SiO₂, 1:10, MeOH: DCM) to yield 19aa as a white

solid (60% yield): ¹H NMR (500 MHz, Methanol d₄) d 8.02 (s, 1H),

7.18 (dd, J = 8.6, 2.5 Hz, 1H), 7.14 (d, J = 2.4 Hz, 1H), 6.67–6.58

(m, 3H), 6.41–6.34 (m, 4H), 6.22–6.13 (m, 2H), 3.99 (t, J = 2.9 Hz,

1H), 3.17 (s, 3H), 3.12 (s, 3H), 2.62 (p, J = 1.6 Hz, 2H), 2.34 (tt, J =

11.7, 4.1 Hz, 1H), 2.20–2.15 (m, 4H), 1.94 (d, J = 2.6 Hz, 3H), 1.88

(t, 4H), 1.62 (s, 3H). ¹³C NMR (126 MHz, MeOD) d 165.63, 159.55,

159.25, 158.92, 156.15, 149.11, 138.27, 130.65, 129.56, 128.81,

127.88, 125.47, 124.67, 121.65, 121.07, 114.89, 112.71, 110.79,

109.74, 69.73, 56.58, 55.46, 54.90, 29.22, 27.49, 22.88, 7.57. HRMS

(ESI+), m/z [M+H⁺] calculated for C₃₃H₃₆N₂O₆571.2682; found

571.2667.

4.3.25. N-(7-(((1r,4r)-4-Benzamidocyclohexyl)oxy)-8-methyl-2-oxo-

2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide

(19ad)

Compound 16a (0.03 g, 0.08 mmol) was dissolved in anhydrous

tetrahydrofuran (0.8 mL). Triethylamine (0.013 mL, 0.08 mmol)

was added and the mixture was cooled to 0 °C. After 10 min, benzyl

chloride (0.013 mL, 0.08 mmol) was added dropwise. The reaction

was quenched after 12 h, via the addition of water. The mixture

was worked-up with ethyl acetate (3 ꢀ 5 mL), dried, concentrated,

and puriﬁed via column chromatography (SiO₂, 1:10, MeOH: DCM)

to afford 19ad as a white amorphous solid (70% yield): ¹H NMR

(500 MHz, Chloroform-d) d 8.78 (s, 1H), 8.70 (s, 1H), 7.90 (d, J =

8.7 Hz, 2H), 7.78–7.73 (m, 2H), 7.49 (d, J = 7.1 Hz, 1H), 7.42 (t, J =

7.5 Hz, 2H), 7.36 (t, J = 7.9 Hz, 1H), 7.30 (d, J = 8.6 Hz, 1H), 7.12

(d, J = 7.7 Hz, 1H), 7.09 (t, J = 2.0 Hz, 1H), 7.06 (d, J = 8.4 Hz, 1H),

6.93 (dd, J = 8.2, 2.5 Hz, 1H), 6.87 (d, J = 8.7 Hz, 1H), 6.10 (d, J =

7.8 Hz, 1H), 4.30 (tt, J = 9.8, 3.5 Hz, 1H), 4.09 (dq, J = 10.5, 3.6 Hz,

1H), 3.87 (d, J = 18.7 Hz, 6H), 2.32 (s, 3H), 2.21 (td, J = 11.5, 10.1,

5.1 Hz, 5H), 1.71 (tdd, J = 13.1, 9.9, 4.0 Hz, 2H), 1.49–1.38 (m,

2H). ¹³C NMR (126 MHz, CDCl₃) d 167.34, 165.86, 160.13, 159.86,

159.69, 157.62, 149.80, 138.97, 135.06, 131.84, 130.32, 129.54,

128.94, 128.49, 127.22, 125.82, 124.66, 122.36, 115.62, 113.50,

111.38, 111.11, 76.44, 56.25, 55.69, 48.25, 30.77, 30.70, 8.74. HRMS

(ESI+), m/z [M+Na⁺] calculated for C₃₈H₃₆N₂O₇Na 655.2420; found

655.2437.

4.3.23. N-(7-(((1r,4r)-4-Acetamidocyclohexyl)oxy)-8-methyl-2-oxo-

2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide

(19ab)

Compound 16a (0.03 g, 0.08 mmol) was dissolved in anhydrous

dichloromethane (0.8 mL). Triethylamine (0.013 mL, 0.08 mmol)

was added and the mixture was cooled to 0 °C. After 10 min, acetic

anhydride (0.008 mL, 0.08 mmol) was added dropwise. The reac-

tion was quenched after 12 h, via the addition of water. The mix-

ture was worked-up with dichloromethane (3 x 5 mL), dried,

concentrated, and puriﬁed via column chromatography (SiO₂,

1:10, MeOH: DCM) to afford 19ab as a white amorphous solid

(70% yield): ¹H NMR (500 MHz, Chloroform-d) d 8.79 (d, J = 1.3

Hz, 1H), 8.70 (s, 1H), 7.94–7.86 (m, 2H), 7.40–7.34 (m, 1H), 7.31

(d, J = 8.6 Hz, 1H), 7.12 (dd, J = 7.6, 1.4 Hz, 1H), 7.10–7.05 (m, 2H),

6.96–6.90 (m, 1H), 6.87 (d, J = 8.6 Hz, 1H), 5.34 (d, J = 7.8 Hz, 1H),

4.31–4.22 (m, 1H), 3.88 (dd, J = 20.0, 1.4 Hz, 6H), 2.31 (d, J = 1.4

Hz, 3H), 2.19–2.05 (m, 4H), 1.98 (d, J = 1.4 Hz, 3H), 1.71–1.57 (m,

4H). ¹³C NMR (126 MHz, CDCl₃) d 169.42, 165.52, 159.77, 159.51,

159.33, 157.25, 138.61, 131.04, 129.96, 129.17, 128.13, 126.10,

125.44, 124.30, 122.00, 121.64, 115.25, 113.15, 111.01, 110.77,

55.89, 55.33, 47.44, 30.42, 30.30, 23.57, 8.37. HRMS (ESI+), m/z

[M+Na⁺] calculated for C₃₃H₃₄N₂O₇Na 593.2264; found 593.2264.

4.3.26. N-(7-(((1r,4r)-4-((Cyclopropylmethyl)amino)cyclohexyl)oxy)-

8-methyl-2-oxo-2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-

carboxamide (19ae)

Cyclopropylmethyl methanesulfonate (0.18 g, 1.2 mmol) was

added to a solution of DIPEA (0.32 mL, 1.8 mmol), 16a (0.528 g,

1.0 mmol), and anhydrous MeCN (10 mL). The mixture was placed

in a sealed microwave safe septum-capped tube, and heated to

120 °C in a microwave synthesizer. After 1 h the reaction was com-

plete and the vial was cooled to rt and quenched with water. The

reaction was worked-up with EtOAc (3 x 20 mL). The organic layers

were combined, dried, concentrated, and puriﬁed via column chro-

matography (SiO₂, 1:10, MeOH: DCM) to afford 19ae as a white

amorphous solid (30% yield): ¹H NMR (500 MHz, Methanol d₄) d

8.68 (d, J = 3.3 Hz, 1H), 7.85 (dd, J = 8.7, 2.6 Hz, 1H), 7.80 (d, J =

2.6 Hz, 1H), 7.33 (d, J = 2.4 Hz, 1H), 7.32–7.29 (m, 2H), 7.06–7.04

(m, 3H), 6.87–6.82 (m, 2H), 4.52 (s, 1H), 4.35–4.30 (m, 1H), 3.81

(d, J = 24.9 Hz, 6H), 2.83 (m, 1H), 2.29 (s, 3H), 2.24 (t, J = 5.5 Hz,

2H), 2.10 (d, J = 13.0 Hz, 3H), 1.94–1.89 (m, 2H), 1.81 (td, J = 11.7,

3.4 Hz, 1H), 1.68 (d, J = 12.2 Hz, 2H) ꢁ0.08 to ꢁ0.16 (m, 1H),

ꢁ0.66 (dt, J = 7.7, 2.7 Hz, 2H), ꢁ0.82 to ꢁ1.01 (m, 2H). ¹³C NMR

4.3.24. N-(7-(((1r,4r)-4-(Benzylamino)cyclohexyl)oxy)-8-methyl-2-

oxo-2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-

carboxamide (19ac)

Benzyl bromide (0.12 mL,1.0 mmol), was added to a solution of

16a (1.32 g, 2.5 mmol), K₂CO₃(0.15 g, 1.1 mmol), in MeCN (30 mL)

and heated at reﬂux. After 24 h. the reaction was cooled to rt,

quenched with water, and worked-up with EtOAc (3 x 50 mL).

The organic layers were combined, dried, concentrated and puri-

ﬁed via column chromatography (SiO₂, 1:10, MeOH: DCM) to

afford 19ac as a white amorphous solid (50% yield): ¹H NMR

(500 MHz, Chloroform-d) d 8.79 (s, 1H), 8.69 (s, 1H), 7.92–7.87

(m, 2H), 7.53 (d, J = 7.4 Hz, 2H), 7.41–7.30 (m, 6H), 7.12 (ddd, J =

7.6, 1.6, 1.0 Hz, 1H), 7.09 (dd, J = 2.6, 1.6 Hz, 1H), 7.06 (d, J = 8.7

Hz, 1H), 6.93 (ddd, J = 8.2, 2.6, 1.0 Hz, 1H), 6.87 (d, J = 8.7 Hz, 1H),

4.32 (ddt, J = 9.5, 6.7, 3.4 Hz, 1H), 3.96 (s, 2H), 3.89 (s, 3H), 3.85

(s, 3H), 2.82 (dd, J = 12.3, 8.8 Hz, 1H), 2.27 (s, 3H), 2.21 (d, J =

11.3 Hz, 5H), 1.71–1.62 (m, 2H), 1.49 (td, J = 11.2, 9.9, 2.6 Hz,

2H).¹³C NMR (126 MHz, CDCl₃) d 165.62, 159.87, 159.64, 159.44,

(126 MHz, MeOD)

d 166.72, 160.53, 160.11, 157.39, 150.36,

139.43, 131.66, 130.08, 129.42, 129.30, 126.45, 125.77, 122.32,

122.06, 116.31, 115.83, 114.33, 113.11, 113.00, 111.67, 110.22,

70.84, 56.59, 55.97, 54.33, 30.53, 28.05, 25.67, 13.36, 8.32, 3.19.

HRMS (ESI+), m/z [M+H⁺] calculated for C₃₅H₃₈N₂O₆583.2808;

found 583.2806.

4.3.27. 3⁰,6-Dimethoxy-N-(8-methyl-2-oxo-7-(((1r,4r)-4-

((tetrahydrofuran-3-yl)amino)cyclohexyl)oxy)-2H-chromen-3-yl)-

[1,1⁰-biphenyl]-3-carboxamide (19ag)

Compound 19ag was obtained following the procedure for the

synthesis of 19ae as a white amorphous solid (30%): ¹H NMR

(500 MHz, Methanol d₄) d 8.72 (d, J = 3.5 Hz, 1H), 7.89 (dd, J = 8.8,

2.5 Hz, 1H), 7.80 (d, J = 2.6 Hz, 1H), 7.33 (d, J = 2.6 Hz, 1H), 7.30–

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L.K. Forsberg et al. / Bioorganic & Medicinal Chemistry xxx (2018) xxx–xxx

11

7.27 (m, 2H), 7.06–7.01 (m, 3H), 6.86–6.80 (m, 2H), 4.55 (s, 1H),

4.38 (t, J = 5.9 Hz, 1H), 4.32–4.29 (m, 1H), 3.88 (q, J = 7.5 Hz, 1H),

3.80 (d, J = 24.7 Hz, 6H), 3.74 (dt, J = 8.3, 4.2 Hz, 1H), 3.67 (dq, J =

12.9, 4.5, 3.8 Hz, 1H), 2.82 (m, 1H), 2.23 (s, 3H), 2.11 (d, J = 13.2

Hz, 3H), 1.99 (ddd, J = 14.2, 9.4, 6.0 Hz, 1H), 1.94–1.88 (m, 2H),

1.83–1.80 (m, 2H), 1.67 (m, 2H). ¹³C NMR (126 MHz, MeOD) d

162.01, 159.37, 159.32, 157.72, 144.53, 131.41, 129.97, 129.47,

129.39, 128.48, 127.32, 125.61, 123.82, 123.02, 122.00, 120.41,

117.85, 115.50, 115.15, 114.22, 75.22, 71.27, 66.62, 58.19, 57.71,

53.29, 52.39, 51.11, 35.02, 31.72, 27.97, 27.69 7.31. HRMS (ESI+),

m/z [M+H⁺] calculated for C₃₅H₃₈N₂O₇599.2709; found 599.2720.

29.07, 27.85, 26.00, 23.10, 8.22, 7.93. HRMS (ESI+), m/z [M+H⁺] cal-

culated for C₃₃H₃₆N₂O₆571.2682; found 571.2688.

4.3.31. N-(7-(((1s,4s)-4-Acetamidocyclohexyl)oxy)-8-methyl-2-oxo-

2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide

(19bb)

Compound 19bb was obtained following the procedure for the

synthesis of 19ab as a white amorphous solid (70%): ¹H NMR

(500 MHz, Chloroform-d) d 8.81 (s, 1H), 8.70 (s, 1H), 7.92 (dd, J =

8.6, 2.4 Hz, 1H), 7.89 (d, J = 2.4 Hz, 1H), 7.38 (d, J = 7.9 Hz, 1H),

7.36–7.32 (m, 1H), 7.13 (ddd, J = 7.6, 1.6, 1.0 Hz, 1H), 7.09 (dd, J

= 2.6, 1.6 Hz, 1H), 7.07 (d, J = 8.7 Hz, 1H), 6.93 (ddd, J = 8.3, 2.6,

1.0 Hz, 1H), 6.87 (d, J = 8.8 Hz, 1H), 5.40 (d, J = 8.1 Hz, 1H), 4.63

(t, J = 3.3 Hz, 1H), 3.90 (s, 3H), 3.86 (s, 3H), 2.36 (s, 3H), 2.11–

2.03 (m, 2H), 2.00 (s, 3H), 1.86–1.81 (m, 2H), 1.78–1.69 (m, 2H),

1.65–1.57 (m, 7H). ¹³C NMR (126 MHz, CDCl₃) d 169.41, 165.64,

159.90, 159.70, 159.45, 156.91, 149.58, 138.74, 131.15, 130.07,

129.33, 128.32, 126.21, 125.80, 124.54, 122.14, 121.70, 115.37,

114.98, 113.39, 113.28, 111.13, 110.28, 71.51, 47.44, 28.74, 27.68,

23.78, 8.56. HRMS (ESI+), m/z [M+Na⁺] calculated for C₃₃H₃₄N₂O₇-

Na 593.2264; found 593.2270.

4.3.28. 3⁰,6-Dimethoxy-N-(8-methyl-2-oxo-7-(((1r,4r)-4-

(((tetrahydrofuran-3-yl)methyl)amino)cyclohexyl)oxy)-2H-chromen-

3-yl)-[1,1⁰-biphenyl]-3-carboxamide (19ah)

Compound 19ah was obtained following the procedure for the

synthesis of 19ae as a white amorphous solid (30%): ¹H NMR

(500 MHz, Methanol d₄) d 7.95 (d, J = 2.9 Hz, 1H), 7.19–7.15 (m,

1H), 7.05 (d, J = 2.3 Hz, 1H), 6.57 (d, J = 2.7 Hz, 1H), 6.55–6.49 (m,

2H), 6.34 (ddt, J = 7.7, 6.5, 2.4 Hz, 3H), 6.29–6.22 (m, 2H), 6.03

(ddd, J = 6.5, 5.5, 3.6 Hz, 1H), 3.62 (s, 1H), 3.54–3.38 (m, 1H), 3.15

(d, J = 1.8 Hz, 3H), 3.11 (d, J = 3.2 Hz, 3H), 2.98–2.86 (m, 1H), 2.77

(td, J = 5.8, 2.9 Hz, 1H), 2.76–2.69 (m, 1H), 2.64–2.55 (m, 1H),

2.52 (s, 3H), 2.49 (dt, J = 3.5, 1.3 Hz, 1H), 2.40 (dd, J = 8.9, 4.5 Hz,

1H), 1.70–1.56 (m, 1H), 1.40 (dd, J = 5.6, 3.4 Hz, 4H), 1.30–1.24

(m, 2H), 1.18 (dddd, J = 17.5, 14.8, 10.3, 4.5 Hz, 2H), 0.99–0.84

(m, 1H). ¹³C NMR (126 MHz, MeOD) d 165.47, 159.47, 158.37,

149.04, 144.19, 138.32, 130.51, 129.49, 128.41, 128.09, 125.73,

123.58, 121.19, 114.11, 112.61, 110.78, 70.48, 67.09, 63.34, 55.35,

54.82, 53.07, 49.15, 41.00, 33.72, 28.08, 7.66. HRMS (ESI+), m/z

[M+H⁺] calculated for C₃₆H₄₀N₂O₇613.2807; found 613.2801.

4.3.32. N-(7-(((1s,4s)-4-(Benzylamino)cyclohexyl)oxy)-8-methyl-2-

oxo-2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-

carboxamide (19bc)

Compound 19bc was obtained following the procedure for the

synthesis of 19ac as a white amorphous solid (70%): ¹H NMR

(500 MHz, Chloroform-d) d 8.79 (d, J = 2.0 Hz, 1H), 8.70 (d, J = 2.9

Hz, 1H), 7.94–7.88 (m, 3H), 7.41–7.36 (m, 3H), 7.35–7.27 (m, 5H),

7.23–7.19 (m, 1H), 7.13 (dt, J = 7.6, 1.2 Hz, 2H), 7.10–7.04 (m,

3H), 6.93 (dd, J = 8.3, 2.5 Hz, 1H), 6.86 (dd, J = 11.6, 8.7 Hz, 1H),

4.66 (s, 0H), 4.60 (dd, J = 7.4, 4.3 Hz, 1H), 3.90 (s, 4H), 3.86 (d, J =

5.3 Hz, 5H), 3.69 (s, 2H), 2.64 (dt, J = 11.5, 3.7 Hz, 1H), 2.36 (d, J =

1.2 Hz, 3H), 2.18–2.02 (m, 2H), 1.79 (ddd, J = 26.3, 13.3, 6.6 Hz,

2H), 1.69–1.55 (m, 6H). ¹³C NMR (126 MHz, CDCl₃) d 165.90,

160.15, 159.73, 149.83, 141.33, 139.02, 131.44, 130.36, 129.57,

128.82, 128.57, 128.49, 127.05, 126.56, 125.89, 124.88, 122.43,

115.64, 113.56, 111.41, 110.62, 72.12, 57.15, 56.29, 55.73, 54.24,

51.40, 29.82, 28.83, 28.43, 22.85, 8.83. HRMS (ESI+), m/z [M+H⁺]

calculated for C₃₈H₃₈N₂O₆619.2808; found 619.2811.

4.3.29. 3⁰,6-Dimethoxy-N-(8-methyl-2-oxo-7-(((1r,4r)-4-

((tetrahydro-2H-pyran-4-yl)amino)cyclohexyl)oxy)-2H-chromen-3-

yl)-[1,1⁰-biphenyl]-3-carboxamide (19ai)

Compound 19ai was obtained following the procedure for the

synthesis of 19ae as a white amorphous solid (30%): ¹H NMR

(500 MHz, Methanol d₄) d 8.59 (s, 1H), 7.75 (dd, J = 8.6, 2.5 Hz,

1H), 7.70 (d, J = 2.4 Hz, 1H), 7.26 (d, J = 2.3 Hz, 1H), 7.22–7.15 (m,

2H), 6.97–6.90 (m, 3H), 6.78–6.70 (m, 2H), 4.62, (s, 1H), 4.55 (t, J

= 3.1 Hz, 1H), 3.77 (dt, J = 11.8, 4.2 Hz, 2H), 3.73 (s, 3H), 3.68 (s,

3H), 3.61 (tt, J = 9.1, 4.2 Hz, 1H), 3.27 (ddd, J = 12.1, 10.3, 2.6 Hz,

2H), 3.05–2.92 (m, 1H), 2.19 (s, 3H), 2.09–1.98 (m, 2H), 1.69 (m,

4H), 1.60–1.47 (m, 2H), 1.37 (dtd, J = 13.6, 9.7, 4.2 Hz, 2H), 1.31

(dd, J = 7.1, 2.3 Hz, 4H). ¹³C NMR (126 MHz, MeOD) d 165.64,

159.50, 158.86, 156.21, 149.07, 138.22, 130.59, 129.50, 128.73,

127.84, 125.43, 124.76, 121.59, 114.83, 112.63, 110.75, 109.73,

79.98, 69.68, 59.67, 55.38, 54.81, 53.05, 34.62, 34.47, 27.46,

24.69, 7.49. HRMS (ESI+), m/z [M+H⁺] calculated for C₃₆H₄₀N₂O₇

613.2867; found 613.2854.

4.3.33. N-(7-(((1s,4s)-4-Benzamidocyclohexyl)oxy)-8-methyl-2-oxo-

2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide

(19bd)

Compound 19bd was obtained following the procedure for the

synthesis of 19ad as a white amorphous solid (70%): ¹H NMR

(500 MHz, Chloroform-d) d 8.80 (s, 1H), 8.70 (s, 1H), 8.10 (dd, J =

8.3, 1.3 Hz, 0H), 7.93–7.87 (m, 2H), 7.81–7.76 (m, 2H), 7.52–7.40

(m, 3H), 7.40–7.30 (m, 2H), 7.14–7.03 (m, 3H), 6.93 (ddd, J = 8.3,

2.6, 0.9 Hz, 1H), 6.88 (d, J = 8.8 Hz, 1H), 6.16 (d, J = 8.0 Hz, 1H),

4.66 (dd, J = 4.7, 2.5 Hz, 1H), 4.17–4.08 (m, 1H), 3.99–3.90 (m,

1H), 3.89 (s, 3H), 3.85 (s, 3H), 2.37 (s, 2H), 2.18–2.07 (m, 2H),

1.99–1.92 (m, 2H), 1.83–1.67 (m, 4H). ¹³C NMR (126 MHz, CDCl₃)

d 170.23, 167.04, 165.63, 159.86, 159.66, 159.41, 156.93, 149.53,

138.70, 134.82, 133.56, 131.58, 131.09, 130.21, 130.03, 129.29,

128.69, 128.53, 128.28, 127.00, 126.16, 125.77, 124.55, 122.11,

121.63, 115.34, 114.96, 113.23, 111.10, 110.27, 71.57, 70.90,

48.54, 47.94, 28.81, 28.57, 27.71, 26.92, 8.56. HRMS (ESI+), m/z

[M+Na⁺] calculated for C₃₈H₃₆N₂O₇Na 655.2420; found 655.2391.

4.3.30. N-(7-(((1s,4s)-4-(Ethylamino)cyclohexyl)oxy)-8-methyl-2-

oxo-2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-

carboxamide (19ba)

Compound 19ba was obtained following the procedure for the

synthesis of 19aa as a white amorphous solid (60%): ¹H NMR

(500 MHz, Chloroform-d) d 9.39 (d, J = 9.9 Hz, 2H), 9.22 (s, 1H),

8.77 (d, J = 10.0 Hz, 1H), 8.67 (s, 1H), 7.92–7.85 (m, 2H), 7.33 (dt,

J = 21.9, 8.1 Hz, 2H), 7.14–7.03 (m, 3H), 6.92 (dd, J = 7.9, 2.5 Hz,

1H), 6.83 (dd, J = 12.0, 8.7 Hz, 1H), 4.68 (d, J = 3.7 Hz, 1H), 4.30 (d,

J = 9.6 Hz, 1H), 3.89 (d, J = 6.3 Hz, 3H), 3.85 (s, 3H), 3.06–2.93 (m,

2H), 2.67 (d, J = 5.1 Hz, 2H), 2.22 (d, J = 15.8 Hz, 1H), 2.11 (s, 3H),

2.04–1.92 (m, 4H), 1.70–1.64 (m, 2H), 1.61 (s, 3H). ¹³C NMR (126

MHz, CDCl₃) d 165.34, 159.65, 159.21, 149.32, 138.49, 130.91,

129.86, 129.05, 128.00, 126.94, 125.91, 125.42, 124.07, 122.21,

121.89, 115.13, 113.03, 110.89, 109.74, 55.75, 55.21, 30.28, 29.60,

4.3.34. N-(7-(((1s,4s)-4-((Cyclopropylmethyl)amino)cyclohexyl)oxy)-

8-methyl-2-oxo-2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-

carboxamide (19be)

Compound 19be was obtained following the procedure for the

synthesis of 19ae as a white amorphous solid (30%): ¹H NMR

(500 MHz, Methanol d₄) d 8.70 (d, J = 3.4 Hz, 1H), 7.86 (dd, J = 8.5,

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L.K. Forsberg et al. / Bioorganic & Medicinal Chemistry xxx (2018) xxx–xxx

2.5 Hz, 1H), 7.81 (d, J = 2.7 Hz, 1H), 7.36 (d, J = 2.6 Hz, 1H), 7.34–

7.27 (m, 2H), 7.08–7.01 (m, 3H), 6.89–6.83 (m, 2H), 4.69–4.62

(m, 1H), 4.49 (s, 1H), 3.82 (d, J = 24.7 Hz, 6H), 3.15–3.06 (m, 1H),

2.30 (s, 3H), 2.25 (t, J = 5.7 Hz, 2H), 2.14 (d, J = 13.3 Hz, 3H), 1.92–

1.83 (m, 2H), 1.79 (td, J = 11.9, 3.2 Hz, 1H), 1.66 (d, J = 12.3 Hz,

2H). ꢁ0.04 to ꢁ0.15 (m, 1H), ꢁ0.64 (dt, J = 7.9, 2.9 Hz, 2H), ꢁ0.85

to ꢁ1.03 (m, 2H). ¹³C NMR (126 MHz, MeOD) d 166.82, 160.71,

160.06, 157.41, 150.25, 139.42, 131.78, 130.68, 129.92, 129.90,

126.60, 125.95, 122.77, 122.14, 116.00, 115.66, 114.11, 113.83,

113.80, 111.92, 110.91, 70.89, 56.56, 55.99, 54.21, 30.40, 28.65,

25.90, 13.81, 8.67, 3.29. HRMS (ESI+), m/z [M+H⁺] calculated for

NMR (126 MHz, MeOD) d 165.64, 159.50, 158.86, 156.21, 149.07,

138.22, 130.59, 129.50, 128.73, 127.84, 125.43, 124.76, 121.59,

114.83, 112.63, 110.75, 109.73, 80.02, 69.68, 59.67, 55.38, 54.81,

53.05, 34.62, 34.47, 27.46, 24.69, 7.49. HRMS (ESI+), m/z [M+H⁺]

calculated for C₃₆H₄₀N₂O₇613.2867; found 613.2854.

4.3.38. N-(7-(((1r,4r)-4-(Bis(cyclopropylmethyl)amino)cyclohexyl)

oxy)-8-methyl-2-oxo-2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-

biphenyl]-3-carboxamide (20ae)

Compound 20ae was obtained following the procedure for the

synthesis of 19ae as a white amorphous solid (30%): ¹H NMR

(500 MHz, Methanol d₄) d 8.69 (d, J = 3.4 Hz, 1H), 7.86 (dd, J = 8.8,

2.3 Hz, 1H), 7.82 (d, J = 2.4 Hz, 1H), 7.35 (d, J = 2.2 Hz, 1H), 7.33–

7.28 (m, 2H), 7.10–7.05 (m, 3H), 6.85–6.82 (m, 2H), 4.32–4.30

(m, 1H), 3.85 (d, J = 25.3 Hz, 6H), 2.80 (m, 1H), 2.25 (s, 3H), 2.20

(m, 4H), 2.07 (d, J = 12.9 Hz, 3H), 1.90–1.85 (m, 2H), 1.80 (td, J =

11.5, 3.1 Hz, 1H), 1.69 (d, J = 12.3 Hz, 2H). ꢁ0.10 to ꢁ0.15 (m,

2H), ꢁ0.69 (m, 4H), ꢁ0.79 to ꢁ0.95 (m, 4H). ¹³C NMR (126 MHz,

MeOD) d 166.39, 160.42, 160.02, 157.76, 150.42, 139.12, 131.81,

130.23, 129.76, 129.21, 126.86, 125.52, 122.27, 122.11, 116.31,

115.96, 114.34, 113.75, 113.04, 111.62, 110.72, 70.44, 56.51,

55.93, 54.34, 30.41, 28.22, 25.54, 13.16, 8.02, 3.91. HRMS (ESI+),

m/z [M+H⁺] calculated for C₃₉H₄₄N₂O₆637.3278; found 637.3266.

C₃₅H₃₈N₂O₆583.2808; found 583.2786.

4.3.35. 3⁰,6-Dimethoxy-N-(8-methyl-2-oxo-7-(((1s,4s)-4-

((tetrahydrofuran-3-yl)amino)cyclohexyl)oxy)-2H-chromen-3-yl)-

[1,1⁰-biphenyl]-3-carboxamide (19bg)

Compound 19bg was obtained following the procedure for the

synthesis of 19ag as a white amorphous solid (30%): ¹H NMR

(500 MHz, Methanol d₄) d 8.80 (d, J = 3.4 Hz, 1H), 7.85 (dd, J = 8.8,

2.6 Hz, 1H), 7.79 (d, J = 2.9 Hz, 1H), 7.49 (d, J = 2.4 Hz, 1H), 7.34–

7.28 (m, 2H), 7.03–7.00 (m, 3H), 6.85–6.82 (m, 2H), 4.70–4.65

(m, 1H), 4.50 (s, 1H), 4.40 (t, J = 5.9 Hz, 1H), 3.88 (q, J = 7.6 Hz,

1H), 3.81 (d, J = 24.5 Hz, 6H), 3.76 (dt, J = 8.4, 4.3 Hz, 1H), 3.62

(dq, J = 12.7, 4.5, 3.7 Hz, 1H), 3.12–3.07 (m, 1H), 2.32 (s, 3H), 2.14

(d, 4H), 1.99 (ddd, J = 14.2, 9.4, 6.0 Hz, 1H), 1.92–1.80 (m, 3H),

1.75 (td, J = 11.7, 3.1 Hz, 1H), 1.68 (d, J = 12.4 Hz, 1H). ¹³C NMR

4.3.39. N-(7-(((1r,4r)-4-(Dimethylamino)cyclohexyl)oxy)-8-methyl-2-

oxo-2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-

carboxamide (20af)

(126 MHz, MeOD)

d 162.11, 159.33, 159.02, 157.76, 144.52,

131.44, 129.90, 129.67, 129.23, 128.67, 127.02, 125.61, 123.22,

123.02, 122.00, 120.11, 117.05, 115.59, 115.45, 114.67, 75.09,

71.32, 66.45, 58.65, 57.71, 53.48, 52.23, 51.16, 35.12, 31.45,

27.22, 27.13 7.33. HRMS (ESI+), m/z [M+H⁺] calculated for

Acetic acid (0.5 mg, 8.5 mmol) was added to a solution of 16a

(0.20 g, 1.7 mmol) in methanol (20 mL). Formaldehyde (0.10 g,

3.4 mmol) and NaBH₃CN (0.23 g, 3.7 mmol) were added and the

mixture was stirred at rt. After 12 h the reaction was diluted with

40 mL of H₂O and then extracted with DCM (3 x 30 mL). The

organic layers were combined, dried, and concentrated. The crude

mixture was puriﬁed by column chromatography (SiO₂, 1:10,

MeOH: DCM) to yield 20af as a white solid (40% yield):¹H NMR

(500 MHz, Chloroform-d) d 8.80 (s, 1H), 8.70 (s, 1H), 7.94–7.87

(m, 2H), 7.39–7.32 (m, 2H), 7.12 (dt, J = 7.6, 1.2 Hz, 1H), 7.10–

7.08 (m, 2H), 7.06 (s, 1H), 6.93 (ddd, J = 8.3, 2.6, 0.9 Hz, 1H), 6.85

(d, J = 8.8 Hz, 1H), 4.70 (t, J = 2.9 Hz, 1H), 3.90 (s, 3H), 3.86 (s,

3H), 3.18–3.03 (m, 1H), 2.69 (s, 6H), 2.35 (s, 3H), 2.31–2.21 (m,

2H), 1.99–1.79 (m, 4H), 1.76–1.62 (m, 2H). ¹³C NMR (126 MHz,

CDCl₃) d 165.66, 159.92, 159.56, 159.45, 156.33, 149.56, 138.71,

131.15, 130.07, 129.32, 128.31, 126.15, 125.87, 124.31, 122.13,

115.37, 113.26, 111.13, 110.18, 70.33, 63.46, 39.57, 28.61, 21.12,

8.54. HRMS (ESI+), m/z [M+H⁺] calculated for C₃₃H₃₆N₂O₆

557.2652; found 557.2641.

C₃₅H₃₈N₂O₇599.2709; found 599.2720.

4.3.36. 3⁰,6-Dimethoxy-N-(8-methyl-2-oxo-7-(((1s,4s)-4-

(((tetrahydrofuran-3-yl)methyl)amino)cyclohexyl)oxy)-2H-chromen-

3-yl)-[1,1⁰-biphenyl]-3-carboxamide (19bh)

Compound 19bh was obtained following the procedure for the

synthesis of 19ah as a white amorphous solid (30%): ¹H NMR (500

MHz, Methanol d₄) d 7.90 (d, J = 2.7 Hz, 1H), 7.09–7.05 (m, 1H),

7.04 (d, J = 2.5 Hz, 1H), 6.56 (d, J = 2.9 Hz, 1H), 6.55–6.46 (m, 2H),

6.34 (ddt, J = 7.9, 6.7, 2.4 Hz, 3H), 6.30–6.22 (m, 2H), 6.09 (ddd, J

= 6.8, 5.3, 3.8 Hz, 1H), 3.52–3.36 (m, 1H), 3.05 (d, J = 1.8 Hz, 3H),

3.01 (d, J = 3.2 Hz, 3H), 2.93–2.86 (m, 1H), 2.76 (td, J = 5.6, 2.7 Hz,

1H), 2.73–2.68 (m, 1H), 2.67–2.58 (m, 1H), 2.54 (d, J = 3.3 Hz,

3H), 2.50 (dt, J = 3.4, 1.6 Hz, 1H), 2.41 (dd, J = 8.8, 4.7 Hz, 1H),

1.66–1.53 (m, 1H), 1.44 (dd, J = 5.8, 3.3 Hz, 4H), 1.32–1.24 (m,

2H), 1.15 (dddd, J = 17.3, 15.0, 10.3, 4.3 Hz, 2H), 0.86–0.67 (m,

1H). ¹³C NMR (126 MHz, MeOD) d 165.57, 159.47, 158.87, 149.07,

144.49, 138.25, 130.54, 129.49, 128.71, 128.09, 125.23, 123.56,

121.59, 114.81, 112.61, 110.72, 70.18, 67.39, 63.50, 55.36, 54.80,

53.27, 49.17, 40.94, 33.70, 28.18, 7.64. HRMS (ESI+), m/z [M+H⁺]

calculated for C₃₆H₄₀N₂O₇613.2807; found 613.2792.

4.3.40. N-(7-(((1r,4r)-4-(Bis(tetrahydrofuran-3-yl)amino)cyclohexyl)

oxy)-8-methyl-2-oxo-2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-

biphenyl]-3-carboxamide (20ag)

Compound 20ag was obtained following the procedure for the

synthesis of 19ae as a white amorphous solid (30%): ¹H NMR

(500 MHz, Methanol d₄) d 8.72 (d, J = 3.4 Hz, 1H), 7.81 (dd, J = 8.9,

2.5 Hz, 1H), 7.79 (d, J = 2.6 Hz, 1H), 7.36 (d, J = 2.5 Hz, 1H), 7.31–

7.29 (m, 2H), 7.09–7.02 (m, 3H), 6.90–6.86 (m, 2H), 4.39–4.37

(m, 2H), 4.32–4.29 (m, 1H), 3.88–3.85 (m, 2H), 3.82 (d, J = 24.8

Hz, 6H), 3.70 (m, 2H), 3.66–3.63 (m, 2H), 2.80 (m, 1H), 2.22 (s,

3H), 2.11–2.09 (m, 4H), 1.97 (m, 1H), 1.92–1.86 (m, 2H), 1.83–

1.80 (m, 4H), 1.69 (m, 2H). ¹³C NMR (126 MHz, MeOD) d 162.31,

159.07, 159.02, 157.79, 144.57, 131.71, 129.99, 129.77, 129.19,

128.28, 127.62, 125.65, 123.02, 123.12, 122.02, 120.31, 117.82,

115.57, 115.35, 114.28, 75.32, 71.07, 66.69, 58.49, 57.71, 53.69,

52.39, 51.31, 35.24, 31.02, 27.67, 27.39 7.55. HRMS (ESI+), m/z

[M+H⁺] calculated for C₃₉H₄₄N₂O₈669.3187; found 669.3200.

4.3.37. 3⁰,6-Dimethoxy-N-(8-methyl-2-oxo-7-(((1s,4s)-4-

((tetrahydro-2H-pyran-4-yl)amino)cyclohexyl)oxy)-2H-chromen-3-

yl)-[1,1⁰-biphenyl]-3-carboxamide (19bi)

Compound 19bi was obtained following the procedure for the

synthesis of 19ai as a white amorphous solid (30%): ¹H NMR

(500 MHz, Methanol d₄) d 8.68 (d, J = 2.8 Hz, 1H), 7.83 (dt, J = 8.5,

3.2 Hz, 1H), 7.79 (t, J = 3.1 Hz, 1H), 7.30 (m, 2H), 7.04–6.97 (m,

4H), 6.81 (ddd, J = 16.9, 8.8, 4.7 Hz, 2H), 4.65 (s, 1H), 4.62–4.56

(m, 1H), 3.85–3.70 (m, 2H), 3.55 (ddd, J = 10.5, 6.6, 3.3 Hz, 2H),

3.25 (dt, J = 3.1, 1.6 Hz, 6H), 3.02 (qd, J = 7.5, 2.9 Hz, 1H), 2.66 (d,

J = 2.6 Hz, 1H), 2.27 (d, J = 3.0 Hz, 3H), 2.14–2.04 (m, 2H), 1.83 (d,

J = 13.0 Hz, 2H), 1.72 (d, J = 12.3 Hz, 2H), 1.62 (dd, J = 14.0, 3.1 Hz,

2H), 1.33 (dt, J = 6.7, 2.2 Hz, 2H), 1.29 (dd, J = 6.7, 2.9 Hz, 2H). ¹³C

Please cite this article in press as: Forsberg L.K., et al. Bioorg. Med. Chem. (2018), https://doi.org/10.1016/j.bmc.2018.04.028

L.K. Forsberg et al. / Bioorganic & Medicinal Chemistry xxx (2018) xxx–xxx

13

4.3.41. N-(7-(((1r,4r)-4-(Bis((tetrahydrofuran-3-yl)methyl)

3.90 (s, 3H), 3.86 (s, 3H), 2.51 (t, J = 11.4 Hz, 1H), 2.43 (s, 7H),

2.36 (s, 3H), 2.23–2.14 (m, 2H), 1.81–1.72 (m, 5H), 1.63 (ddt, J =

14.0, 11.1, 2.7 Hz, 2H). ¹³C NMR (126 MHz, CDCl₃) d 165.64,

159.88, 159.68, 159.45, 156.86, 149.60, 138.73, 131.13, 130.08,

129.32, 128.29, 126.23, 125.71, 124.52, 122.14, 115.35, 113.28,

111.11, 110.29, 71.46, 62.92, 40.99, 28.94, 22.62, 8.50. HRMS (ESI

+), m/z [M+H⁺] calculated for C₃₃H₃₆N₂O₆557.2652; found

557.2652.

amino)cyclohexyl)oxy)-8-methyl-2-oxo-2H-chromen-3-yl)-3⁰,6-

dimethoxy-[1,1⁰-biphenyl]-3-carboxamide (20ah)

Compound 20ah was obtained following the procedure for the

synthesis of 19ae as a white amorphous solid (30%): ¹H NMR

(500 MHz, Methanol d₄) d 7.91 (d, J = 2.4 Hz, 1H), 7.11–7.06 (m,

1H), 7.02 (d, J = 2.4 Hz, 1H), 6.60 (d, J = 3.0 Hz, 1H), 6.55–6.46 (m,

2H), 6.35 (ddt, J = 8.1, 6.7, 2.2 Hz, 3H), 6.29–6.21 (m, 2H), 6.00

(ddd, J = 7.0, 5.1, 3.2 Hz, 1H), 3.56–3.42 (m, 1H), 3.17 (d, J = 1.6

Hz, 3H), 3.11 (d, J = 3.0 Hz, 3H), 2.99–2.88 (m, 2H), 2.79 (m, 2H),

2.75–2.68 (m, 2H), 2.63–2.58 (m, 2H), 2.50 (s, 3H), 2.48 (dt, J =

3.3, 1.5 Hz, 1H), 2.30 (m, 2H), 1.88–1.76 (m, 1H), 1.62–1.57 (m,

4H), 1.42–1.30 (m, 2H), 1.25 (m, 2H), 1.09–1.02 (m, 1H). ¹³C NMR

4.3.45. N-(7-(((1s,4s)-4-(Bis(tetrahydrofuran-3-yl)amino)cyclohexyl)

oxy)-8-methyl-2-oxo-2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-

biphenyl]-3-carboxamide (20bg)

Compound 20bg was obtained following the procedure for the

synthesis of 19ae as a white amorphous solid (30%): ¹H NMR

(500 MHz, Methanol d₄) d 8.82 (d, J = 3.5 Hz, 1H), 7.80 (dd, J = 8.6,

2.3 Hz, 1H), 7.75 (d, J = 2.6 Hz, 1H), 7.50 (d, J = 2.3 Hz, 1H), 7.34–

7.28 (m, 2H), 7.05–7.01 (m, 3H), 6.87–6.82 (m, 2H), 4.68–4.63

(m, 1H), 4.42–4.38 (m, 2H), 3.90–3.87 (m, 2H), 3.79 (d, J = 24.8

Hz, 6H), 3.75 (m, 2H), 3.65–3.60 (m, 2H), 3.11–3.06 (m, 1H), 2.28

(s, 3H), 2.10 (d, 4H), 1.99–1.93 (m, 2H), 1.89–1.80 (m, 5H), 1.72

(m, 1H), 1.66 (m, 1H). ¹³C NMR (126 MHz, MeOD) d 162.45,

159.35, 159.02, 157.17, 144.02, 131.86, 129.95, 129.45, 129.23,

128.37, 127.14, 125.01, 123.28, 123.02, 122.09, 120.55, 117.85,

115.51, 115.21, 114.67, 75.34, 71.31, 66.49, 58.62, 57.51, 53.66,

52.12, 51.14, 35.54, 31.13, 27.56, 27.23 7.55. HRMS (ESI+), m/z

[M+H⁺] calculated for C₃₉H₄₄N₂O₈669.3187; found 669.3165.

(126 MHz, MeOD)

d 165.06, 159.63, 158.41, 149.33, 144.21,

138.74, 130.39, 129.36, 128.23, 128.07, 125.56, 123.06, 121.59,

114.61, 112.31, 110.98, 70.08, 67.90, 63.52, 55.06, 54.00, 53.17,

49.37, 40.99, 33.76, 28.48, 7.04. HRMS (ESI+), m/z [M+H⁺] calcu-

lated for C₄₁H₄₈N₂O₈697.3464; found 697.3487.

4.3.42. N-(7-(((1r,4r)-4-(Bis(tetrahydro-2H-pyran-4-yl)

amino)cyclohexyl)oxy)-8-methyl-2-oxo-2H-chromen-3-yl)-3⁰,6-

dimethoxy-[1,1⁰-biphenyl]-3-carboxamide (20ai)

Compound 20ai was obtained following the procedure for the

synthesis of 19ae as a white amorphous solid (30%): ¹H NMR

(500 MHz, Methanol d₄) d 8.50 (s, 1H), 7.78 (dd, J = 8.4, 2.2 Hz,

1H), 7.71 (d, J = 2.4 Hz, 1H), 7.30 (d, J = 2.4 Hz, 1H), 7.23–7.17 (m,

2H), 6.90–6.84 (m, 3H), 6.76–6.70 (m, 2H), 4.50 (t, J = 3.2 Hz, 1H),

3.78 (dt, J = 11.9, 4.0 Hz, 4H), 3.70 (s, 3H), 3.67 (s, 3H), 3.60 (tt, J

= 9.1, 4.2 Hz, 2H), 3.30 (ddd, J = 12.0, 10.1, 2.5 Hz, 4H), 3.05–2.92

(m, 1H), 2.10 (s, 3H), 2.02–1.96 (m, 2H), 1.65 (m, 4H), 1.58–1.49

(m, 2H), 1.39 (dtd, J = 13.5, 9.9, 4.2 Hz, 4H) 1.31 (dd, J = 7.1, 2.3

Hz, 4H). ¹³C NMR (126 MHz, MeOD) d 165.94, 159.55, 158.06,

155.71, 149.00, 138.42, 130.51, 129.00, 128.33, 127.14, 125.44,

124.56, 121.50, 114.89, 112.03, 110.55, 109.43, 79.90, 69.68,

59.63, 55.18, 54.21, 53.15, 34.64, 34.40, 27.56, 24.60, 7.89. HRMS

(ESI+), m/z [M+H⁺] calculated for C₄₁H₄₈N₂O₈697.3403; found

697.3421.

4.3.46. N-(7-(((1s,4s)-4-(Bis((tetrahydrofuran-3-yl)methyl)

amino)cyclohexyl)oxy)-8-methyl-2-oxo-2H-chromen-3-yl)-3⁰,6-

dimethoxy-[1,1⁰-biphenyl]-3-carboxamide (20bh)

Compound 20bh was obtained following the procedure for the

synthesis of 19ae as a white amorphous solid (30%): ¹H NMR (500

MHz, Methanol d₄) d 7.93 (d, J = 2.7 Hz, 1H), 7.20–7.15 (m, 1H),

7.10 (d, J = 2.5 Hz, 1H), 6.66 (d, J = 3.0 Hz, 1H), 6.55–6.40 (m, 2H),

6.30 (ddt, J = 7.7, 6.3, 2.4 Hz, 3H), 6.21–6.17 (m, 2H), 6.09 (ddd, J

= 6.4, 5.1, 3.8 Hz, 1H), 3.55–3.40 (m, 1H), 3.10 (s, 3H), 3.06 (s,

3H), 2.95–2.89 (m, 2H), 2.77 (m, 2H), 2.73–2.68 (m, 2H), 2.60–

2.53 (m, 2H), 2.45(s, 3H), 2.30 (dt, J = 3.3, 1.8 Hz, 1H), 2.22 (m,

2H), 1.88–1.65 (m, 1H), 1.58 (m, 4H), 1.39–1.28 (m, 2H), 1.22 (m,

2H), 1.13–1.06 (m, 1H). ¹³C NMR (126 MHz, MeOD) d 165.53,

159.41, 158.89, 149.05, 144.40, 138.22, 130.44, 129.78, 128.69,

128.19, 125.56, 123.93, 121.88, 114.23, 112.15, 110.63, 70.78,

67.22, 63.42, 55.06, 54.10, 53.57, 49.27, 40.99, 33.82, 28.58, 7.66.

HRMS (ESI+), m/z [M+H⁺] calculated for C₄₁H₄₈N₂O₈697.3464;

found 697.3436.

4.3.43. N-(7-(((1s,4s)-4-(Bis(cyclopropylmethyl)amino)cyclohexyl)

oxy)-8-methyl-2-oxo-2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-

biphenyl]-3-carboxamide (10be)

Compound 10be was obtained following the procedure for the

synthesis of 19ae as a white amorphous solid (30%): ¹H NMR

(500 MHz, Methanol d₄) d 8.75 (d, J = 3.6 Hz, 1H), 7.82 (dd, J = 8.7,

2.4 Hz, 1H), 7.79 (d, J = 2.6 Hz, 1H), 7.35 (d, J = 2.7 Hz, 1H), 7.33–

7.29 (m, 2H), 7.07–7.02 (m, 3H), 6.88–6.82 (m, 2H), 4.66–4.63

(m, 1H), 3.84 (d, J = 24.6 Hz, 6H), 3.13–3.09 (m, 1H), 2.32 (s, 3H),

2.27 (m, 4H), 2.18 (d, J = 13.4 Hz, 3H), 1.93–1.88 (m, 2H), 1.81

(td, J = 12.1, 3.3 Hz, 1H), 1.68 (d, J = 12.4 Hz, 2H). ꢁ0.04 to ꢁ0.14

(m, 2H), ꢁ0.65 (m, 4H), ꢁ0.83 to ꢁ1.00 (m, 4H). ¹³C NMR (126

MHz, MeOD) d 166.20, 160.15, 160.46, 157.42, 150.45, 139.48,

131.38, 130.69, 129.95, 129.00, 126.64, 125.75, 122.74, 122.34,

116.02, 115.36, 114.51, 113.13, 113.00, 111.52, 110.11, 70.49,

56.66, 55.75, 54.81, 30.32, 28.75, 25.91, 13.11, 8.07, 3.55. HRMS

(ESI+), m/z [M+H⁺] calculated for C₃₉H₄₄N₂O₆637.3278; found

637.3292.

4.3.47. N-(7-(((1s,4s)-4-(Bis(tetrahydro-2H-pyran-4-yl)

amino)cyclohexyl)oxy)-8-methyl-2-oxo-2H-chromen-3-yl)-3⁰,6-

dimethoxy-[1,1⁰-biphenyl]-3-carboxamide (20bi)

Compound 20bi was obtained following the procedure for the

synthesis of 19ae as a white amorphous solid (30%): ¹H NMR

(500 MHz, Methanol d₄) d 8.66 (d, J = 2.8 Hz, 1H), 7.82 (dt, J = 8.6,

3.0 Hz, 1H), 7.77 (t, J = 2.8 Hz, 1H), 7.28 (m, 2H), 7.01–6.92 (m,

4H), 6.80 (ddd, J = 16.7, 8.9, 4.5 Hz, 2H), 4.60–4.51 (m, 1H), 3.83–

3.72 (m, 4H), 3.50 (ddd, J = 10.7, 6.6, 3.1 Hz, 4H), 3.27 (d, 6H),

3.02 (qd, J = 7.5, 2.9 Hz, 2H), 2.62 (d, J = 2.8 Hz, 1H), 2.20 (s, 3H),

2.13–2.08 (m, 2H), 1.80 (d, J = 12.8 Hz, 2H), 1.75 (d, J = 12.5 Hz,

2H), 1.60 (dd, J = 14.2, 3.0 Hz, 2H), 1.35 (dt, J = 6.5, 2.4 Hz, 4H),

4.3.44. N-(7-(((1s,4s)-4-(Dimethylamino)cyclohexyl)oxy)-8-methyl-

2-oxo-2H-chromen-3-yl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-

carboxamide (20bf)

Compound 20bf was obtained following the procedure for the

synthesis of 5.7a as a white amorphous solid (60%):¹H NMR (500

MHz, Chloroform-d) d 8.80 (s, 1H), 8.70 (s, 1H), 7.94–7.87 (m,

2H), 7.40–7.30 (m, 2H), 7.13 (ddd, J = 7.6, 1.6, 1.0 Hz, 1H), 7.09

(dd, J = 2.6, 1.6 Hz, 1H), 7.07 (d, J = 8.6 Hz, 1H), 6.93 (ddd, J = 8.3,

2.6, 0.9 Hz, 1H), 6.86 (d, J = 8.7 Hz, 1H), 4.65 (t, J = 3.0 Hz, 1H),

1.30 (dd, J = 6.9, 2.6 Hz, 4H). ¹³C NMR (126 MHz, MeOD)

d

165.04, 159.51, 158.82, 156.22, 149.27, 138.52, 130.50, 129.51,

128.70, 127.14, 125.42, 124.26, 121.50, 114.84, 112.63, 110.25,

109.53, 80.12, 69.08, 59.47, 55.08, 54.82, 53.05, 34.22, 34.44,

27.66, 24.09, 7.59. HRMS (ESI+), m/z [M+H⁺] calculated for

C₄₁H₄₈N₂O₈697.3403; found 697.3390.

Please cite this article in press as: Forsberg L.K., et al. Bioorg. Med. Chem. (2018), https://doi.org/10.1016/j.bmc.2018.04.028

14

L.K. Forsberg et al. / Bioorganic & Medicinal Chemistry xxx (2018) xxx–xxx

4.3.48. 3⁰,6-Dimethoxy-N-((1r,4r)-4-(4-(piperidin-3-yloxy)

12.3, 8.4, 3.4 Hz, 2H), 2.47 (tt, J = 12.1, 3.4 Hz, 1H), 2.30 (t, J =

10.0 Hz, 1H), 2.25–2.18 (m, 2H), 2.10–2.02 (m, 2H), 1.98–1.88 (m,

2H), 1.75 (dtd, J = 12.0, 8.0, 3.7 Hz, 2H), 1.62 (qd, J = 13.2, 3.3 Hz,

2H), 1.36 (qd, J = 12.6, 3.5 Hz, 2H); ¹³C NMR (126 MHz, CDCl₃) d

166.32, 159.42, 159.01, 155.57, 139.26, 139.20, 130.52, 129.39,

129.24, 128.31, 127.84 (2C), 127.36, 122.14, 116.13 (2C), 115.49,

112.91, 110.98, 72.45, 55.94, 55.48, 48.83, 43.35 (2C), 42.86,

33.79 (2C), 33.34 (2C), 31.57 (2C); HRMS (ESI+) m/z [M+H] calcu-

lated for C₃₂H₃₉N₂O₄: 515.2910, found 515.2893.

phenyl)cyclohexyl)-[1,1⁰-biphenyl]-3-carboxamide (26a)

¹H NMR (500 MHz, CDCl₃) d 7.81 (dd, J = 8.6, 2.3 Hz, 1H), 7.67 (d,

J = 2.4 Hz, 1H), 7.35 (t, J = 7.9 Hz, 1H), 7.13–7.09 (m, 3H), 7.07 (dd, J

= 2.6, 1.6 Hz, 1H), 7.00 (d, J = 8.7 Hz, 1H), 6.91 (ddd, J = 8.3, 2.6, 1.0

Hz, 1H), 6.88 (d, J = 8.7 Hz, 2H), 5.93 (d, J = 8.2 Hz, 1H), 4.35 (tt, J =

7.0, 3.4 Hz, 1H), 4.04 (tdt, J = 11.5, 7.7, 3.8 Hz, 1H), 3.86 (s, 3H), 3.85

(s, 3H), 3.23 (dd, J = 12.6, 2.2 Hz, 1H), 3.04 (d, J = 12.9 Hz, 1H), 3.00–

2.83 (m, 2H), 2.46 (tt, J = 12.1, 3.4 Hz, 1H), 2.24–2.18 (m, 2H), 2.13–

2.04 (m, 1H), 2.04–1.72 (m, 4H), 1.72–1.51 (m, 4H), 1.43–1.31 (m,

2H); ¹³C NMR (126 MHz, CDCl₃) d 166.33, 159.42, 159.00, 155.49,

139.42, 139.19, 130.51, 129.40, 129.23, 128.30, 127.87 (2C),

127.35, 122.13, 116.06 (2C), 115.48, 112.91, 110.97, 71.42, 55.93,

55.47, 49.76, 48.83, 45.74, 42.86, 33.77 (2C), 33.32 (2C), 29.31,

4.3.52. N-((1r,4r)-4-(4-((1-(2-Hydroxyethyl)piperidin-4-yl)oxy)

phenyl)cyclohexyl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide

(26e)

¹H NMR (600 MHz, CDCl₃) d 7.81 (dd, J = 8.6, 2.4 Hz, 1H), 7.68 (d,

J = 2.4 Hz, 1H), 7.35 (t, J = 7.9 Hz, 1H), 7.14–7.09 (m, 3H), 7.07 (t, J =

2.1 Hz, 1H), 7.01 (d, J = 8.6 Hz, 1H), 6.91 (ddd, J = 8.3, 2.6, 0.9 Hz,

1H), 6.84 (d, J = 8.6 Hz, 2H), 5.91 (d, J = 8.1 Hz, 1H), 4.32 (bs, 1H),

4.04 (tdt, J = 11.8, 8.0, 4.0 Hz, 1H), 3.86 (s, 3H), 3.85 (s, 3H), 3.63

(t, J = 5.4 Hz, 2H), 2.81 (bs, 2H), 2.59 (t, J = 5.4 Hz, 2H), 2.51–2.37

(m, 3H), 2.25–2.16 (m, 3H), 2.05–1.98 (m, 2H), 1.95 (d, J = 12.7

Hz, 2H), 1.89–1.78 (m, 2H), 1.62 (qd, J = 13.2, 3.3 Hz, 2H), 1.36

(qd, J = 12.6, 3.4 Hz, 2H); ¹³C NMR (151 MHz, CDCl₃) d 166.34,

159.45, 159.04, 155.76, 139.21, 139.19, 130.56, 129.40, 129.23,

128.30, 127.81 (2C), 127.39, 122.15, 116.15 (2C), 115.52, 112.93,

111.01, 72.22, 59.39, 57.95, 55.94, 55.48, 50.43 (2C), 48.86, 42.87,

33.80 (2C), 33.35 (2C), 30.90 (2C); HRMS (ESI+) m/z [M+H] calcu-

lated for C₃₄H₄₃N₂O₅: 559.3172, found 559.3090.

22.85; HRMS (ESI+) m/z [M+H] calculated for

C₃₂H₃₉N₂O₄:

515.2910, found 515.2887.

4.3.49. N-((1r,4r)-4-(4-((1-(2-Hydroxyethyl)piperidin-3-yl)oxy)

phenyl)cyclohexyl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide

(26b)

¹H NMR (600 MHz, CDCl₃) d 7.81 (dd, J = 8.6, 2.3 Hz, 1H), 7.68 (d,

J = 2.3 Hz, 1H), 7.35 (t, J = 7.9 Hz, 1H), 7.13–7.09 (m, 3H), 7.07 (t, J =

2.1 Hz, 1H), 7.01 (d, J = 8.6 Hz, 1H), 6.91 (dt, J = 8.3, 1.6 Hz, 1H),

6.85 (d, J = 8.6 Hz, 2H), 5.91 (d, J = 8.1 Hz, 1H), 4.31 (tt, J = 8.6,

4.0 Hz, 1H), 4.04 (tdt, J = 11.8, 8.0, 4.0 Hz, 1H), 3.86 (s, 3H), 3.85

(s, 3H), 3.61 (t, J = 5.4 Hz, 2H), 3.06 (d, J = 11.0 Hz, 1H), 2.76 (d, J

= 11.1 Hz, 1H), 2.58 (t, J = 5.4 Hz, 2H), 2.47 (tt, J = 12.2, 3.5 Hz,

1H), 2.26 (t, J = 9.8 Hz, 1H), 2.24–2.16 (m, 4H), 2.10–2.04 (m, 1H),

1.98–1.91 (m, 2H), 1.83 (dp, J = 12.6, 4.2 Hz, 1H), 1.67–1.56 (m,

3H), 1.54–1.45 (m, 1H), 1.36 (qd, J = 12.6, 3.4 Hz, 2H); ¹³C NMR

4.3.53. 3⁰,6-Dimethoxy-N-((1r,4r)-4-(4-((1-(2-methoxyethyl)

piperidin-4-yl)oxy)phenyl)cyclohexyl)-[1,1⁰-biphenyl]-3-carboxamide

(26f)

(151 MHz, CDCl₃)

d 166.33, 159.45, 159.03, 155.90, 139.29,

139.22, 130.56, 129.41, 129.23, 128.30, 127.85 (2C), 127.41,

122.15, 116.02 (2C), 115.51, 112.94, 111.01, 73.07, 59.40, 57.99,

57.83, 55.94, 55.48, 53.28, 48.86, 42.88, 33.81 (2C), 33.33 (2C),

30.21, 23.32; HRMS (ESI+) m/z [M+H] calculated for C₃₄H₄₃N₂O₅:

559.3172, found 559.3150.

¹H NMR (500 MHz, CDCl₃) d 7.81 (dd, J = 8.6, 2.4 Hz, 1H), 7.67 (d,

J = 2.4 Hz, 1H), 7.35 (t, J = 7.9 Hz, 1H), 7.14 (d, J = 8.7 Hz, 2H), 7.10

(dt, J = 7.6, 1.2 Hz, 1H), 7.07 (dd, J = 2.7, 1.6 Hz, 1H), 7.01 (d, J =

8.6 Hz, 1H), 6.91 (ddd, J = 8.3, 2.6, 1.0 Hz, 1H), 6.83 (d, J = 8.7 Hz,

2H), 5.91 (d, J = 8.0 Hz, 1H), 4.55 (s, 1H), 4.04 (tdt, J = 11.2, 7.4,

3.7 Hz, 1H), 3.86 (s, 3H), 3.85 (s, 3H), 3.37 (s, 3H), 3.28–2.93 (m,

8H), 2.57–2.41 (m, 3H), 2.25–2.17 (m, 2H), 2.10 (d, J = 14.3 Hz,

2H), 1.97–1.88 (m, 2H), 1.62 (qd, J = 13.2, 3.3 Hz, 2H), 1.42–1.32

(m, 2H); ¹³C NMR (126 MHz, CDCl₃) d 166.36, 159.46, 159.06,

154.88, 140.00, 139.21, 130.57, 129.40, 129.24, 128.32, 128.08

(2C), 127.35, 122.15, 116.03 (2C), 115.54, 112.92, 111.03, 77.36,

67.96, 59.11, 57.50, 55.94, 55.48, 49.53 (2C), 48.82, 42.87, 33.74

(2C), 33.32, (2C) 27.79 (2C); HRMS (ESI+) m/z [M+H] calculated

for C₃₅H₄₅N₂O₅: 573.3328, found 573.3301.

4.3.50. 3⁰,6-Dimethoxy-N-((1r,4r)-4-(4-((1-(2-methoxyethyl)

piperidin-3-yl)oxy)phenyl)cyclohexyl)-[1,1⁰-biphenyl]-3-carboxamide

(26c)

¹H NMR (500 MHz, CDCl₃) d 7.81 (dd, J = 8.6, 2.3 Hz, 1H), 7.67 (d,

J = 2.4 Hz, 1H), 7.39–7.29 (m, 1H), 7.13–7.09 (m, 3H), 7.07 (dd, J =

2.6, 1.5 Hz, 1H), 7.01 (d, J = 8.7 Hz, 1H), 6.91 (ddd, J = 8.2, 2.6, 0.9

Hz, 1H), 6.86 (d, J = 8.6 Hz, 2H), 5.88 (d, J = 8.1 Hz, 1H), 4.36 (bs,

1H), 4.04 (dtd, J = 11.7, 7.8, 4.0 Hz, 1H), 3.86 (s, 3H), 3.85 (s, 3H),

3.52 (bs, 2H), 3.34 (s, 3H), 3.14 (bs, 1H), 2.82 (bs, 1H), 2.63 (bs,

2H), 2.46 (tt, J = 12.1, 3.5 Hz, 1H), 2.26–2.17 (m, 3H), 2.19–2.01

(m, 2H), 1.95 (d, J = 13.3 Hz, 2H), 1.85–1.75 (m, 1H), 1.70–1.52

(m, 4H), 1.36 (qd, J = 12.7, 3.5 Hz, 2H); ¹³C NMR (126 MHz, CDCl₃)

d 166.34, 159.46, 159.04, 155.34, 139.23, 139.10, 130.58, 129.41,

129.24, 128.30, 127.80 (2C), 127.43, 122.16, 115.92 (2C), 115.52,

112.96, 111.02, 73.04, 59.06, 58.50, 58.09, 55.95, 55.49, 54.04,

53.91, 48.87, 42.89, 33.83 (2C), 33.35 (2C), 30.25, 23.32; HRMS

(ESI+) m/z [M+H] calculated for C₃₅H₄₅N₂O₅: 573.3328, found

573.3325.

4.3.54. N-((1R,4r)-4-(4-(((1s,4S)-4-Aminocyclohexyl)oxy)

phenyl)cyclohexyl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide

(26g)

Compound 5.18b was obtained following the procedure for the

synthesis of 16a as a white amorphous solid (70%): ¹H NMR (500

MHz, Methylene Chloride-d2) d 7.78 (ddd, J = 8.7, 6.2, 2.5 Hz, 1H),

7.69 (t, J = 2.9 Hz, 1H), 7.32 (t, J = 7.9 Hz, 1H), 7.15–7.10 (m, 2H),

7.05 (td, J = 9.0, 6.2 Hz, 2H), 6.89 (dd, J = 8.3, 2.6 Hz, 1H), 6.84–

6.78 (m, 2H), 4.26 (t, J = 3.8 Hz, 1H), 4.17 (dd, J = 9.7, 4.6 Hz, 1H),

3.94 (ddt, J = 11.4, 7.5, 4.3 Hz, 1H), 3.85 (d, J = 2.6 Hz, 3H), 3.82

(d, J = 2.3 Hz, 3H), 3.13–3.07 (m, 1H), 2.52–2.42 (m, 1H), 2.23–

2.05 (m, 5H), 2.01–1.96 (m, 1H), 1.94–1.88 (m, 1H), 1.83–1.73

(m, 1H), 1.71–1.35 (m, 8H). ¹³C NMR (126 MHz, CD₂Cl₂) d 159.53,

159.50, 155.78, 139.81, 139.36, 130.51, 129.56, 129.16, 127.85,

122.11, 116.21, 115.51, 112.88, 111.05, 74.40, 55.85, 55.40, 42.87,

33.41, 29.65, 29.60, 29.12, 28.49. HRMS (ESI+), m/z [M+H⁺] calcu-

lated for C₃₃H₄₀N₂O₄529.3076; found 529.3056.

4.3.51. 3⁰,6-Dimethoxy-N-((1r,4r)-4-(4-(piperidin-4-yloxy)

phenyl)cyclohexyl)-[1,1⁰-biphenyl]-3-carboxamide (26d)

¹H NMR (500 MHz, CDCl₃) d 7.82 (dd, J = 8.6, 2.4 Hz, 1H), 7.68 (d,

J = 2.4 Hz, 1H), 7.38–7.31 (m, 1H), 7.14–7.09 (m, 3H), 7.07 (dd, J =

2.6, 1.6 Hz, 1H), 7.01 (d, J = 8.7 Hz, 1H), 6.92 (ddd, J = 8.2, 2.6, 0.9

Hz, 1H), 6.85 (d, J = 8.7 Hz, 2H), 5.90 (d, J = 8.1 Hz, 1H), 4.38 (tt, J

= 7.6, 3.6 Hz, 1H), 4.04 (tdt, J = 10.7, 6.9, 3.5 Hz, 1H), 3.86 (s, 3H),

3.85 (s, 3H), 3.19 (ddd, J = 12.7, 6.9, 3.7 Hz, 2H), 2.83 (ddd, J =

Please cite this article in press as: Forsberg L.K., et al. Bioorg. Med. Chem. (2018), https://doi.org/10.1016/j.bmc.2018.04.028

L.K. Forsberg et al. / Bioorganic & Medicinal Chemistry xxx (2018) xxx–xxx

15

4.3.55. N-((1R,4r)-4-(4-(((1r,4R)-4-Aminocyclohexyl)oxy)

phenyl)cyclohexyl)-3⁰,6-dimethoxy-[1,1⁰-biphenyl]-3-carboxamide.

(26h)

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