Development of Novel Mitochondrial Pyruvate Carrier Inhibitors to Treat Hair Loss

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Development of Novel Mitochondrial Pyruvate Carrier Inhibitors to

Treat Hair Loss

Xiaoguang Liu, Aimee A. Flores, Lisa Situ, Wen Gu, Hui Ding, Heather R. Christofk, William E. Lowry,

and Michael E. Jung *

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ABSTRACT: Herein, we report the synthesis and evaluation of novel analogues of UK-5099 both in vitro and in vivo for the

development of mitochondrial pyruvate carrier (MPC) inhibitors to treat hair loss. A comprehensive understanding of the

structure−activity relationship was obtained by varying four positions of the hit compound, namely, the alkyl group on the N1

position, substituents on the indole core, various aromatic and heteroaromatic core structures, and various Michael acceptors. The

major discovery was that the inhibitors with a 3,5-bis(triﬂuoromethyl)benzyl group at the N1 position were shown to have much

better activity than JXL001 (UK-5099) to increase cellular lactate production. Additionally, analogue JXL069, possessing a 7-

azaindole heterocycle, was also shown to have signiﬁcant MPC inhibition activity, which further increases the chemical space for

drug design. Finally, more than 10 analogues were tested on shaved mice by topical treatment and promoted obvious hair growth on

mice.

activate, hair loss occurs. Our collaborators, Lowry, Christofk,

and co-workers, discovered that the activation of HFSCs can

be achieved by stimulating the activity of lactate dehydrogen-

ase (LDH), the enzyme catalyzing the reduction of pyruvate to

lactate.⁷Genetic deletion of mitochondrial pyruvate carrier

(MPC) or pharmacological inhibition of MPC via treatment

with UK-5099 resulted in increased LDH activity in HFSCs.⁷

When the MPC is knocked down or pharmacologically

inhibited, pyruvate in the cytoplasm cannot enter the

mitochondria and instead is converted to other metabolites,

such as lactate by lactate dehydrogenase (LDH), which results

in the increase of LDH activity and the activation of HFSCs.

Overall, our collaborators discovered a novel mechanism to

INTRODUCTION

■

Hair loss, also known as alopecia or baldness, is a very

common health problem. Fifty percent of males and 25% of

females by age 50 suﬀer pattern hair loss.¹Common types of

hair loss include male-pattern hair loss, female-pattern hair

loss, alopecia areata, and a thinning of hair known as telogen

eﬄuvium.¹Minoxidil (Rogaine) and ﬁnasteride (Propecia) are

the two Food and Drug Administration (FDA)-approved

medicines to treat hair loss but only 30−40% of patients

experience hair growth with the requirement of repeated

treatment, and serious side eﬀects have been reported.²

Surgical procedures to treat baldness are expensive and can

be painful. Therefore, a better therapy for hair loss is highly

desired.

Hair follicle growth occurs in cycles. Each cycle consists of a

long growing phase (anagen), a short transitional phase

(catagen), and a short resting phase (telogen).³The ability of

the hair follicles to maintain this cycle depends on the presence

of hair follicle stem cells (HFSCs).⁴The HFSCs are typically

inactive but can quickly “wake up” and actively divide when a

new hair growth cycle is initiated.^5,6When HFSCs fail to

Received: September 8, 2020

Published: February 3, 2021

https://dx.doi.org/10.1021/acs.jmedchem.0c01570

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promote hair growth and MPC has been proven to be a

promising therapeutic target for the treatment of hair loss.

MPC, located on the inner mitochondrial membrane, is a

transporter protein that transports pyruvate from the

cytoplasm to the mitochondria. Although great eﬀorts have

been taken to study MPC, its molecular structure is still not

clear.^8,9The MPC protein family consists of three members,

MPC1, MPC2, and MPC3. The MPC1−MPC2 complex, an

oligomeric structure of about 150 kD, is an essential

component of the mitochondrial pyruvate carrier in yeast,

ﬂies, and mammals.⁹UK-5099 is a known covalent inhibitor of

MPC, which reversibly reacts with a cysteine on the protein

with a K_ivalue of 0.05 μM.¹⁰UK-5099 has been reported to

promote lactate production rate as a result of MPC inhibition

in various settings.^11,12Our collaborators, Lowry, Christofk,

and co-workers, demonstrated that the topical treatment of

mice in the telogen phase with UK-5099 (20 μM) led to a

robust acceleration of the hair cycle.⁷Therefore, UK-5099 is a

good hit compound for developing novel MPC inhibitors as a

treatment of hair loss.

Herein, we describe an extensive structure−activity relation-

ship (SAR) study of a series of (E)-2-cyano-3-(1H-indol-3-

yl)acrylic acids and their structural analogues. We have

developed a general synthetic route to these compounds that

allows one to prepare many analogues rapidly. All of these new

compounds have been tested for their ability to promote

cellular lactate production in vitro. The compounds that

enhanced cellular lactate production rate were then tested on

mice topically for their ability to accelerate hair growth. We

have identiﬁed more than 10 compounds with potent MPC

inhibitory activity and the ability to promote hair growth in

mice.

Table 1. Analogues Synthesized by Following Scheme 1

analogue

R¹

R²

R³

JXL001

JXL002

JXL003

JXL004

JXL005

JXL006

JXL007

JXL012

JXL013

JXL014

JXL026

JXL027

JXL028

JXL029

JXL035

JXL093

H

6-Cl

H

6-Cl

H

4-Cl

6-F

7-Cl

5-F

4-Cl

OH

OEt

OH

OEt

Ph

H

Ph

2-OMeC₆H₄

4-OMeC₆H₄

Ph

2-OMeC₆H₄

Ph

Scheme 2. Synthesis of JXL020

RESULTS AND DISCUSSION

■

Chemistry. The synthesis of the analogues, which are

labeled JXL001−JXL096, was carried out as shown in the

various schemes. The synthesis of JXL001 is shown in Scheme

1. Copper-mediated N-arylation of indole-3-carboxaldehyde 1

Scheme 1. Synthesis of JXL001

with 3,5-bis(triﬂuoromethyl)benzyl bromide generated com-

pound 3 in good yield. Knoevenagel condensation with ethyl

2-cyanoacetate provided JXL011. Finally, aqueous LiOH-

mediated saponiﬁcation aﬀorded analogue JXL020. When

substituted indole-3-carboxaldehydes and various benzyl

bromides were used, various analogues were synthesized

rapidly by following the same synthetic route as for JXL020.

The analogues are listed in Table 2. Based on the result of our

SAR study, the analogues of JXL020 with the 3,5-bis-

(triﬂuoromethyl)benzyl group at the N1 position were

shown to have much better activity than JXL001 in promoting

cellular lactate production. A series of analogues with a 3,5-

bis(triﬂuoromethyl)benzyl group at the N1 position were

with iodobenzene followed by Knoevenagel condensation with

ethyl 2-cyanoacetate provided the ester version of UK-5099,

JXL004. Finally, aqueous LiOH-mediated saponiﬁcation

delivered the desired compound JXL001. Following the

same synthetic route with substituted indole-3-carboxalde-

hydes and substituted iodobenzenes, various analogues were

synthesized rapidly (Table 1).

Analogue JXL020 was synthesized by the synthetic route

shown in Scheme 2. Benzylation of indole-3-carboxaldehyde 1

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Grignard reagent to carbon dioxide followed by diisobutyla-

luminum hydride (DIBAL) reduction and mesylation.

Nucleophilic substitution of compound 7 with indole 8

generated compound 9. Upon treatment with triﬂuoroacetic

acid, the tert-butyl group was cleaved to form the desired

analogue JXL079. The amide analogue JXL080 was synthe-

sized by an N,N’-dicyclohexylcarbodiimide (DCC) coupling

reaction of JXL003 and compound 6 (Scheme 3b).

Table 2. Analogues Synthesized by Following Scheme 2

In addition to analogues with the indole core, analogues with

other aromatic and heteroaromatic cores were also synthesized.

4-Pyridinecarboxaldehyde and substituted benzaldehydes were

used to synthesize analogues following a synthetic route similar

to that used for JXL001 (Scheme 1). The analogues are listed

in Table 4. Some analogues with nitrogen-containing hetero-

cycles were also synthesized using a synthetic route similar to

that used for JXL069 in Scheme 4. Benzylation of 7-azaindole

4 with KOH as the base generated compound 5 with good

yield and regioselectivity. The Vilsmeier−Haack reaction

followed by Knoevenagel condensation with ethyl 2-

cyanoacetate provided JXL082. Finally, aqueous LiOH-

mediated saponiﬁcation furnished the analogue JXL069.

Analogues with a 4-azaindole, a 6-azaindole, and a substituted

7-azaindole core were successfully synthesized and are listed in

Table 5. The Vilsmeier−Haack reaction failed to generate the

3-carboxyaldehydes of 4-azaindole and 6-azaindole, so the

formylation was carried out using hexamethylenetetramine

(HMTA) in acetic acid (Duﬀ reaction).

analogue

R¹

R²

R⁴

JXL008

JXL009

JXL010

JXL011

JXL015

JXL016

JXL017

JXL018

JXL019

JXL020

JXL036

JXL037

JXL039

JXL040

H

5-F

6-F

OEt

OH

OEt

OH

4-F

3,4-di-F

3,5-di-F

3,5-bis(CF₃)

4-F

3,4-di-F

3,5-di-F

H

3,5-bis(CF₃)

3,4-di-F

3,5-di-F

3,4-di-F

3,5-di-F

The analogues with substituents at the 3-position of the

acrylates, JXL083, JXL084, and JXL085, were synthesized, as

shown in Scheme 5. Aldehyde 3 was oxidized by a Pinnick

oxidation to the carboxylic acid 10, which was further

converted to acyl chloride 11. Deprotonation of ethyl 2-

cyanoacetate by lithium diisopropylamide (LDA) and

nucleophilic addition to compound 11 generated analogue

JXL083. Analogue JXL084 was synthesized by acetylation of

JXL083 and JXL085 was synthesized by chlorination of

JXL083.

synthesized by following a similar synthetic route as shown for

JXL020 (Table 3).

The analogue JXL079 with a substituted homobenzyl group

at the N1 position was synthesized as shown in Scheme 3a.

The homobenzyl mesylate 7 was synthesized from the

commercially available compound 3,5-bis(triﬂuoromethyl)-

benzyl bromide by nucleophilic addition of its derived

Table 3. Analogues with 3,5-Bis(triﬂuoromethyl)benzyl

Group at the N1 Position

Analogues with various Michael acceptors were synthesized

by Knoevenagel condensations (Figure 1). The bis-nitrile

JXL021 was synthesized by the condensation with malononi-

trile and JXL025 was synthesized by the condensation with

diethyl malonate followed by LiOH-mediated monosaponiﬁ-

cation. JXL086 and JXL095 were generated by the

condensation of aldehydes 3 and 12, respectively, with diethyl

(cyanomethyl)phosphonate. JXL096 was synthesized from

JXL086 by trimethylsilyl bromide (TMSBr)-mediated cleavage

of the ethyl groups. Analogues with thiazolidinedione (TZD)

structures were synthesized similarly as JXL024. Condensation

of various aldehydes with either thiazolidine-2,4-dione or 2-

iminothiazolidin-4-one in reﬂux acetic acid aﬀorded the

analogues listed in Figure 2.

Biological Evaluation: SAR Analysis. When the MPC is

inhibited, pyruvate in the cytoplasm cannot enter the

mitochondria and instead is converted to other metabolites,

such as lactate by lactate dehydrogenase (LDH). Since our

collaborators previously found that LDH activity correlated

with hair follicle stem cell activation, and genetic deletion of

LDH activity via LDHa knockout in hair follicle stem cells

blocked their activation,⁷we were particularly interested in

using rerouting of pyruvate toward lactate production as a

readout for MPC inhibition in this study. The use of cell-based

lactate production as a readout of MPC inhibition was a

practical ﬁrst-level readout for screening new compounds, but

analogue

R¹

5-F

6-F

4-Cl

4-Br

4-F

7-F

5-Cl

4-CN

4-CO₂H

4-OBn

6-OBn

7-OBn

4-OMe

5-Br

R²

analogue

R¹

6-Br

6-Cl

7-Cl

4-Br

4-F

H

5-Cl

4-CN

4-CO₂Me

H

R²

JXL038

JXL041

JXL050

JXL051

JXL052

JXL053

JXL054

JXL055

JXL056

JXL057

JXL058

JXL059

JXL060

JXL061

OH

JXL062

JXL063

JXL064

JXL065

JXL066

JXL068

JXL078

JXL081

JXL089

JXL090

JXL091

JXL092

JXL094

OH

OEt

Ot-Bu

OMe

morpholine

OEt

OH

NH₂

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Scheme 3. Synthesis of JXL079 and JXL080

promote cellular lactate production. The data of the in vitro

test is shown in Figure 3.

Table 4. Analogues with Benzene Core and JXL022

Here, we used the (E)-2-cyano-3-(1-phenyl-1H-indol-3-

yl)acrylic acid (JXL001) as a reference compound and de-

signed new compounds to vary the four most easily altered

positions, namely, (1) the alkyl group at N1, (2) the

substituents on the indole core, (3) various aromatic and

heteroaromatic cores, and (4) various Michael acceptors. Our

goal was to identify candidates for the treatment of hair loss

with signiﬁcant improvement in potency.

Analogues at the N1 Position (R3). We envisioned that

modifying the N-phenyl group would be a good starting point

for conducting an SAR by medicinal chemistry. Several N-aryl-

substituted analogues are listed in Table 1. Analogue JXL002

without the phenyl group showed little inhibitory eﬀect on

MPC, implying that the phenyl group is important for UK-

5099’s biological activity. Similarly, compound JXL003 was

worse than JXL004 in terms of stimulating lactate production.

Analogues with N-2-methoxyphenyl and N-4-methoxyphenyl

groups (JXL006 and JXL007, esters; JXL014 and JXL012,

carboxylic acids) showed similar or slightly better activity than

JXL001 to promote the cellular lactate production rate. Due to

the harsh reaction conditions and low reaction yields for the N-

arylation step, we switched our attention to analogues with N-

benzyl groups.

analogue

R⁵

analogue

R⁵

JXL030

JXL032

JXL034

JXL042

JXL043

JXL044

2-F

4-F

JXL045

JXL046

JXL047

JXL048

JXL049

3-CF₃, 4-CF₃

2-Cl, 3-CF₃

3-CF₃, 4-F

H

4-CF₃

3-F, 4-Me

3-F, 4-F

2-F, 4-F

4-OH

that it is certainly not synonymous with MPC inhibition. Our

hit compound, UK-5099, is known to be a covalent inhibitor.

To conﬁrm its mechanism of action, the analogue of UK-5099

without the alkene double bond was synthesized and tested in

the cell-based assay. The reduced UK-5099 showed no eﬀect

on lactate production rate, meaning that it did not inhibit

MPC (data not shown). This result supports that UK-5099 is a

covalent inhibitor.

A dose−response experiment of the commercial UK-5099

and in-house-prepared JXL001 with MCF10A cells was

conducted to conﬁrm their inhibition of MPC by measuring

the cells’ lactate production rate (data not shown). This result

indicated that both samples promoted lactate production in a

dose-dependent pattern. Because both samples provided the

highest lactate production rate at 10 μM and the same

concentration of UK-5099 was used in the earlier in vivo study

to detect an eﬀect on hair growth,⁷a concentration of 10 μM

was chosen for the evaluation of the activity of the analogues to

We then investigated the N-benzyl analogues because the

homologated analogues possess high structural similarity with

JXL001. A series of analogues with diﬀerent benzyl groups

were synthesized and tested (Table 2). Although most of these

analogues showed better activity in increasing lactate

production rate, the analogue with the 3,5-bis-

(triﬂuoromethyl)benzyl group, JXL020, was shown to have

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Scheme 4. Synthesis of JXL069

production. For chloro-substituted analogues (JXL050,

JXL054, JXL063, and JXL064), only the 4-Cl analogue

(JXL050) showed better activity than JXL020 to increase the

lactate production rate. Similarly, the analogue with 4-Br

(JXL051) was better than JXL020, while analogues with 5-Br

and 6-Br were worse than JXL020. Among the three

benzyloxy-substituted indole analogues (JXL057, JXL058,

and JXL059), only the 7-OBn analogue JXL059 showed

better activity than JXL020 but the 4-OBn analogue failed to

increase its biological activity in promoting lactate production

rate, probably due to the steric eﬀect of the large substituent.

Based on the analysis, more analogues with small substituents

at the 4-position of indole were synthesized and evaluated. The

analogue with 4-CN (JXL055) was better than JXL020 and

the analogue with 4-OMe (JXL060) was similar to JXL020 in

terms of promoting lactate production. The analogue with 4-

COOH (JXL056) showed much worse activity than JXL020

in inhibition of MPC, perhaps due to its high polarity that may

make it hard to penetrate the cell membrane. Based on this

data, it seems that large substituents on the indole ring are

harmful to the inhibitory activity for MPC and small

substituents at the four-position are beneﬁcial for the desired

activity.

Table 5. Analogues with Azaindole Cores

analogue

X

Y

Z

R¹

R²

JXL069

JXL072

JXL073

JXL076

JXL077

JXL082

JXL087

JXL088

C

N

C

CH

N

CH

N

H

N/A

H

Cl

Br

H

Cl

Br

OH

OEt

CH

N

Analogues with Aromatic and Heteroaromatic Cores

Other than the Indole Core. Analogues with 4-pyridine and

many substituted benzene cores (Table 4) were shown to have

poorer activity than JXL001 in promoting cellular lactate

production, implying that the indole core structure is

important for the biological activity.

the highest ability to inhibit MPC. JXL079, the homologue of

JXL020, was shown to have lower potency than JXL020 in

inhibition of MPC. Additionally, the amide JXL080 did not

promote the cellular lactate production. Up to this point, the

3,5-bis(triﬂuoromethyl)benzyl group was the best moiety at

the N1 position of the indole core.

We further evaluated other aromatic cores to identify novel

MPC inhibitors (Table 5). Analogues containing azaindole

cores were synthesized according to Scheme 4. To our delight,

the 7-azaindole analogue (JXL069) was much more potent

than the 4-azaindole analogue (JXL072), the 6-azaindole

analogue (JXL073), and the hit compound JXL020 with an

indole core. The improved inhibitory activity may result from a

new ligand−protein interaction or the electronic eﬀect of the

azaindole structure. The 4-Cl-7-azaindole and 4-Br-7-azaindole

Analogues with Substituents on the Indole Ring.

Taking JXL020 as the new hit compound, the inﬂuence of

substituents on the indole core was explored (the analogues

are listed in Table 3). Comparing the four analogues

containing 4-F, 5-F, 6-F, and 7-F (JXL052, JXL038, JXL041,

and JXL053) with JXL020, JXL052 (the 4-F analogue) and

JXL053 (the 7-F analogue) showed better activity than

JXL020 to increase the lactate production rate, while the

other two analogues could hardly promote the lactate

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Scheme 5. Synthesis of JXL083, JXL084, and JXL085

Figure 1. Analogues with various Michael acceptors.

Figure 2. Analogues containing the TZD group.

analogues (JXL076 and JXL077) showed similar activity as

JXL020 in promoting lactate production rate. This new

azaindole structure broadened the chemical space for further

SAR study.

Analogues with Various Michael Acceptors. We

envisioned that manipulating the Michael acceptor moiety

might be important for the potency and selectivity of the

analogues. Modiﬁcation of this group can change the

molecule’s lipophilicity and electrophilicity. To test this idea,

analogues with diﬀerent alkyl groups were synthesized (Table

3). The methyl ester (JXL078) and the tert-butyl ester

analogues (JXL068) showed a dramatic loss of potency

compared with the corresponding carboxylic acid analogue

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Figure 3. In vitro evaluation of analogues of JXL001−JXL096. Human breast epithelial (MCF10A) cells were treated with 10 μM analogue for 24−

30 h, and the cellular lactate production rate was measured, relative to vehicle (dimethyl sulfoxide, DMSO) treatment and normalized to cell

number. Data were compiled from sequential batches of experiments, each with their own DMSO controls and JXL001 and JXL020 treatments for

comparison. Error bars indicate +1 s.d. Representative experiments are shown, with most analogues tested at least twice. Shading indicates adjusted

p-values calculated by performing ordinary one-way analysis of variance (ANOVA) followed by Dunnett’s post-test, comparing each analogue to

DMSO.

(JXL020), while the ethyl ester (JXL011) showed similar

potency as JXL020. Based on this discovery, many other ethyl

ester analogues were tested. To our delight, most of them were

shown to have similar potency to their corresponding

carboxylic acid analogues, such as JXL065 and JXL051,

JXL066 and JXL052, and JXL082 and JXL069.

fueled OCR assay we used on permeabilized cells for

measuring IC₅₀of our MPC inhibitors is the most direct

functional assay available for measuring MPC inhibition. In

this assay, the only substrate for mitochondrial respiration

given to the permeabilized cells is pyruvate. The MPC

inhibitors block the pyruvate-fueled OCRs in a dose-depend-

ent manner. We were able to convince ourselves that these

values were direct since our MPC inhibitors had no eﬀect on

succinate-fueled OCR, suggesting a direct eﬀect on the MPC

and not some other component of the electron transport chain.

Analogues JXL001, JXL020, and JXL069 were tested for their

ability to inhibit the pyruvate-fueled respiration of permeabi-

lized cells, and IC₅₀values were determined (Figure 4).

JXL001 showed an IC₅₀value of around 140 nM. JXL020 and

JXL069 showed much lower IC₅₀values (16.6 and 42.8 nM)

than JXL001, which implies that they are more potent than

JXL001 in inhibiting MPC.

In Vivo Evaluation of MPC Inhibitors. To determine the

eﬃcacy of the compounds on hair growth, C57BL/6J mice

were shaved at postnatal day 50 during the telogen phase of

the hair cycle. Compounds were suspended in lotion and

applied topically to the shaved back skin of the mice every

other day for 2 weeks. As seen in Figure 5, all of the analogues

that showed the ability to promote lactate production in the in

vitro assay were also able to stimulate hair growth over the

course of 2 weeks. JXL020, the analogue with 3,5-bis-

(triﬂuoromethyl)benzyl group at the N1 position, strongly

promoted hair growth. The four-substituted analogues,

JXL035, JXL050, JXL051, and JXL052, were also shown to

have a strong ability to accelerate hair growth on mice. This

data showed that most of the MPC inhibitors active in vitro

could promote hair growth in mice. We tried to compare the in

vivo eﬃcacy of the MPC inhibitors with diﬀerent in vitro

The analogues with malononitrile (JXL021) and dicarbox-

ylate (JXL025) were shown to have poorer activity than

JXL001 in increasing the lactate production rate (Figure 1).

Analogues containing the TZD groups in Figure 2 failed to

show promising activity in promoting lactate production.

Amide analogues (JXL081 and JXL094) showed very strong

ability to promote lactate production; however, substantial cell

death was observed (data was not shown). The indole core-

based diethyl phosphonate analogue (JXL086; Figure 1)

promoted lactate production more than JXL001. However, the

corresponding phosphonic acid analogue (JXL096) failed to

promote lactate production rate, perhaps due to its poor cell

permeability. The 7-azaindole core-based diethyl phosphonate

analogue (JXL095) showed even stronger potency than

JXL086 in promoting lactate production. Overall, adjusting

the Michael acceptors showed a dramatic eﬀect on the

biological activity.

IC₅₀Determination for Several Analogues. The test of

mitochondrial pyruvate uptake would directly conﬁrm MPC

inhibition. However, given the structural similarity between

UK-5099 (which does not inhibit pyruvate carboxylase (PC)

or pyruvate dehydrogenase (PDH) activity) and the new

analogues, it is reasonable that decreased pyruvate-driven

respiration is a result of MPC inhibition. Since the MPC

enables pyruvate-fueled respiration, measuring how well the

UK-5099 analogues block the oxygen consumption rate

(OCR) can be an assay of MPC inhibition.¹³The pyruvate-

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four sites around the core structure and tested the activity of

those analogues in promoting cellular lactate production to

establish a structure−activity relationship for this system. The

substituent at the N1 position is crucial for the inhibition of

MPC. After screening various aryl and substituted benzyl

groups, the analogue containing a 3,5-bis(triﬂuoromethyl)-

benzyl group at the N1 position, JXL020, was shown to have

much better ability than JXL001 to promote cellular lactate

production in vitro (IC₅₀= 16.6 nM) and accelerate hair

growth on mice. Modiﬁcation of substituents on the indole

core structure revealed that small substituents, such as F, Cl,

Br, and CN, at the C4 position were beneﬁcial for the

biological activity. In addition, analogues with diﬀerent

aromatic and heteroaromatic core structures were synthesized

and tested. Analogue JXL069, having a 7-azaindole hetero-

cycle, was also shown to have signiﬁcantly better activity than

the analogues containing a 4-azaindole or a 6-azaindole core

structure. This discovery further increases the chemical space

for the development of MPC inhibitors. Furthermore, diﬀerent

types of Michael acceptors were synthesized and tested for

their ability to increase lactate production rate. Analogues with

the ethyl ester functional group were shown to have similar

activity to inhibit MPC as their corresponding carboxylic acid

analogues, such as ester JXL011 and acid JXL020. This ﬁnding

might provide more opportunities in drug formulation since

the esters have diﬀerent solubilities than the carboxylic acids.

Analogues containing amides (JXL081 and JXL094) and

diethyl phosphonates (JXL086 and JXL095) showed potent

MPC inhibition. Finally, more than 10 analogues were tested

on mice topically to promote hair growth. The analogues

containing the 3,5-bis(triﬂuoromethyl)benzyl group at the N1

position (JXL020, JXL050, JXL051, and JXL052) stimulated

hair growth dramatically. With the novel and potent MPC

inhibitors in hand, the pharmacokinetic and safety tests of

these compounds are ongoing with the ﬁnal goal of developing

drugs to treat hair growth.

Figure 4. IC₅₀measurement of JXL001, JXL020, and JXL069.

MCF10A cells were treated with analogues to measure oxygen

consumption rate (OCR) under diﬀerent conditions. State 3 is the

measurement of respiration in permeabilized cells in the presence of

pyruvate, malate, and ADP. IC₅₀values are indicated in bold. Error

bars denote 1 standard error of mean (SEM; n = 2).

potencies on the MPC inhibition. However, the hair growth

assay is not very quantitative with respect to timing. All of the

MPC inhibitors perform better than the controls, but it is

diﬃcult to run enough animals together to see statistically

signiﬁcant data to distinguish between the activities of the

various MPC inhibitors.

CONCLUSIONS

■

This study reports the synthesis and evaluation of novel

analogues of UK-5099 both in vitro and in vivo for the

development of MPC inhibitors to treat hair loss. We modiﬁed

Figure 5. Evaluation of analogues in vivo. On each row, the mice were treated at the same time with the corresponding MPC inhibitors. Animals

treated topically with the analogue (20 μM) show pigmentation and hair growth, indicative of the entry into anagen, after 8 days of treatment. Full

anagen, indicated by a full coat of hair, is achieved after 14 days of treatment. Mice treated topically with vehicle control do not show pigmentation

nor hair growth even after 12 days of treatment.

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Experimental Details for the Synthesis of JXL020. To the

solution of indole-3-carboxaldehyde (3 mmol, 435 mg) in dry DMF

(6 mL) were added 3,5-bis(triﬂuoromethyl)benzyl bromide (1.2

equiv, 3.6 mmol, 660 μL) and KOH (1.2 equiv, 3.6 mmol, 200 mg) at

0 °C. The reaction mixture was stirred at 21 °C for 2 h. After the

reaction completion shown by TLC, water (6 mL) was added into the

reaction vial. The reaction mixture was extracted by dichloromethane

(15 mL × 3). The combined organic layer was dried by sodium

sulfate and concentrated. The residue was puriﬁed by ﬂash column

chromatography (hexanes/ethyl acetate = 8:1) to provide the desired

product. Yield: 90%, 1001.7 mg.

To the solution of 1-(3,5-bis(triﬂuoromethyl)benzyl)-1H-indole-3-

carbaldehyde (1 mmol, 371 mg) in ethanol (1 mL) were added ethyl

2-cyanoacetate (1.3 equiv, 1.3 mmol, 140 μL) and ^L-proline (40 mol

%, 0.4 mmol, 58 mg). The reaction was stirred at 21 °C for 12 h and a

yellow solid precipitated gradually. After completion of the reaction,

ice-cold water (2 mL) was added into the reaction vial. The solid was

separated by Buchner funnel ﬁltration and washed with water (2 mL

× 3) and dried to aﬀord the desired product. Yield: 93%, 433 mg.

To the solution of (E)-ethyl 3-(1-(3,5-bis(triﬂuoromethyl)benzyl)-

1H-indol-3-yl)-2-cyanoacrylate (0.21 mmol, 100 mg) in THF (2 mL)

was added 0.5 N LiOH solution (3 equiv, 0.4 mmol, 0.8 mL). The

reaction mixture was stirred at 21 °C for 1 h. After the reaction

completion shown by TLC, THF was evaporated. Concentrated HCl

was added dropwise to acidify the reaction mixture until pH was

lower than 1; meanwhile, a yellow solid precipitated. Ice-cold water (5

mL) was added to the reaction mixture, and the solid was separated

by Buchner funnel ﬁltration and washed with water (5 mL × 3). After

drying by vacuum, the solid was washed with 2 mL of solvent mixture

(hexanes/ethyl acetate = 5:1) 5−10 times and monitored by TLC

until nonpolar impurities disappear (the nonpolar compound was the

retro-aldol condensation product, which can be recovered from the

ﬁltrate). Finally, the purity of the product was checked by NMR.

Yield: 55%, 52 mg.

The following compounds were synthesized by a route similar to

that described for JXL020: JXL008, JXL009, JXL010, JXL011,

JXL015, JXL016, JXL017, JXL018, JXL019, JXL036, JXL037,

JXL038, JXL039, JXL040, JXL041, JXL050, JXL051, JXL052,

JXL053, JXL054, JXL055, JXL56, JXL057, JXL058, JXL059,

JXL060, JXL061, JXL062, JXL063, JXL064, JXL065, JXL066,

JXL068, JXL078, JXL081, JXL089, JXL090, and JXL091.

Experimental Details for the Synthesis of JXL024. To the

solution of 1-phenyl-1H-indole-3-carbaldehyde (0.4 mmol, 90 mg) in

AcOH (3 mL) were added thiazolidine-2,4-dione (1 equiv, 0.4 mmol,

46.8 mg) and NaOAc (3 equiv, 98 mg). The reaction mixture was

stirred at reﬂux for 24 h. After it was cooled to 21 °C, the reaction

mixture was ﬁltered by vacuum ﬁltration and washed with AcOH (3

mL × 3) and water (5 mL × 3). After drying by vacuum, the desired

product was produced. Yield: 34%, 44 mg.

EXPERIMENTAL SECTION

■

General. Toluene was distilled from sodium under an argon

atmosphere. Dichloromethane was distilled from calcium hydride

under an argon atmosphere. All other solvents or reagents were

puriﬁed according to literature procedures. H NMR spectra were

recorded on Bruker spectrometers at 500 MHz and are reported

1

relative to deuterated solvent signals. Data for H NMR spectra are

reported as follows: chemical shift (δ ppm), multiplicity, coupling

constant (Hz), and integration. Splitting patterns are designated as

follows: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; and

br, broad. ¹³C NMR spectra were recorded on Bruker spectrometers

at 100 MHz. Data for ¹³C NMR spectra are reported as follows:

chemical shift (δ ppm), multiplicity, and coupling constant (Hz).

1

Splitting patterns are designated the same way as in H NMR. High-

resolution mass spectrometry (HRMS) was taken on a Waters LCT

Premier with ACQUITY UPLC with an autosampler. Mass spectra

were recorded from 70 to 2000 Da. The purity of compounds was

analyzed with an Agilent 1200 HPLC system equipped with a

G1312A binary pump, a G1314A autosampler, a G1314A VWD, and

an Agilent SB-Aq C18 column (1.8 μm, 4.6 × 50 mm²). Mass spectra

were recorded using an Agilent 6130 LC/MS system equipped with

an ESI source. Method: solution A: H₂O w/0.1% formic acid and

solution B: ACN w/0.1% formic acid; increase of B from 10 to 99%,

0.5−2 min; 99%, 2−17 min; ﬂow rate 1.0 mL/min; UV 254 nm. The

purity of all ﬁnal compounds was determined to be >95% by analytical

high-performance liquid chromatography (HPLC) analysis except the

compounds JXL024, JXL025, and JXL056 with around 90% purity.

All solvents were liquid chromatography-tandem mass spectrometry

(LC−MS/MS) grade and purchased from Fisher Scientiﬁc.

Experimental Details for the Synthesis of JXL001 and Related

Analogues. To the solution of indole-3-carboxaldehyde (2.8 mmol,

411 mg) in dry dimethylformamide (DMF; 6 mL) were added Cu₂O

(0.3 equiv, 0.84 mmol, 120 mg), K₂CO₃(2.0 equiv, 5.6 mmol, 774

mg), and iodobenzene (2.0 equiv, 5.6 mmol, 624 μL) sequentially.

The reaction was stirred and reﬂuxed for 24 h, at which point thin-

layer chromatography (TLC) indicated that the reaction was

completed. After it was cooled to 21 °C, the reaction mixture was

ﬁltrated through a celite pad eluting with ethyl acetate. The ﬁltrate

was washed with saturated NaCl solution, and the organic phase was

dried by sodium sulfate and concentrated. The residue was puriﬁed by

ﬂash column chromatography (hexanes/ethyl acetate = 8:1) to

provide the desired product. Yield: 89%, 550.7 mg.

To the solution of 1-phenyl-indole-3-carbaldehyde (1 mmol, 221

mg) in ethanol (1 mL) were added ethyl 2-cyanoacetate (1.3 equiv,

1.3 mmol, 140 μL) and ^L-proline (40 mol %, 0.4 mmol, 58 mg). The

reaction was stirred at 21 °C for 12 h and a yellow solid precipitated

gradually. After completion of the reaction, ice-cold water (2 mL) was

added into the reaction vial. The solid was separated by Buchner

funnel ﬁltration and washed with water (2 mL × 3) and dried to

aﬀord the desired product. Yield: 95%, 300 mg.

To the solution of (E)-ethyl 2-cyano-3-(1-phenyl-1H-indol-3-

yl)acrylate (0.32 mmol, 100 mg) in tetrahydrofuran (THF; 2 mL)

was added 0.5 N LiOH solution (3 equiv, 0.6 mmol, 1.2 mL). The

reaction mixture was stirred at 21 °C for 1 h. After reaction

completion shown by TLC, THF was evaporated. Concentrated HCl

was added dropwise to acidify the reaction mixture until the pH was

lower than 1; meanwhile, a yellow solid precipitated. Ice-cold water (5

mL) was added to the reaction mixture, and the solid was separated

by Buchner funnel ﬁltration and washed with water (5 mL × 3). After

drying by vacuum, the solid was washed with 2 mL of solvent mixture

(hexanes/ethyl acetate = 5:1) 5−10 times and monitored by TLC

until nonpolar impurities disappear (the nonpolar compound was the

retro-Aldol condensation product, which can be recovered from the

ﬁltrate). Finally, the purity of the product was checked by NMR.

Yield: 65%, 60 mg.

The following compounds were synthesized by a route similar to

that described for JXL024: JXL023, JXL067, JXL070, JXL072,

JXL074, and JXL075.

Experimental Details for the Synthesis of JXL022. To the

solution of 4-pyridinecarboxaldehyde (1 mmol, 107 mg) in ethanol (1

mL) were added ethyl 2-cyanoacetate (1.3 equiv, 1.3 mmol, 140 μL)

and L-proline (40 mol %, 0.4 mmol, 58 mg). The reaction was stirred

at 21 °C for 12 h, and a yellow solid precipitated gradually. After

completion of the reaction, ice-cold water (2 mL) was added into the

reaction vial. The solid was separated by Buchner funnel ﬁltration and

washed with water (2 mL × 3) and dried to aﬀord the desired

product, ethyl (E)-2-cyano-3-(pyridin-4-yl)acrylate, which was used

for the next step without further puriﬁcation.

To the solution of (E)-2-cyano-3-(pyridin-4-yl)acrylate (0.21

mmol, 42.4 mg) in THF (2 mL) was added 0.5 N LiOH solution

(3 equiv, 0.4 mmol, 0.8 mL). The reaction mixture was stirred at 21

°C for 1 h. After the reaction completion shown by TLC, THF was

evaporated. Concentrated HCl was added dropwise to acidify the

reaction mixture until pH was lower than 1; meanwhile, a yellow solid

precipitated. Ice-cold water (5 mL) was added to the reaction

mixture, and the solid was separated by Buchner funnel ﬁltration and

The following compounds were synthesized by a route similar to

that described for JXL001: JXL002, JXL003, JXL004, JXL005,

JXL006, JXL007, JXL012, JXL013, JXL014, JXL021, JXL025,

JXL026, JXL027, JXL028, JXL029, JXL035, and JXL093.

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washed with water (5 mL × 3). After drying by vacuum, the solid was

washed with 2 mL of solvent mixture (hexanes/ethyl acetate = 5:1)

5−10 times and monitored by TLC until nonpolar impurities

disappear. Finally, the purity of the product was checked by NMR.

Yield: 64%, 23.4 mg.

The following compounds were synthesized by a route similar to

that described for JXL022: JXL030, JXL031, JXL032, JXL033,

JXL034, JXL042, JXL43, JXL044, JXL045, JXL046, JXL047,

JXL048, and JXL049.

Experimental Details for the Synthesis of JXL069. To the

solution of 7-azaindole 4 (1 mmol, 118 mg) in dry DMF (2 mL) were

added 3,5-bis(triﬂuoromethyl)benzyl bromide (1.2 equiv, 1.2 mmol,

220 μL) and KOH (1.2 equiv, 1.2 mmol, 67 mg) at 0 °C. The

reaction mixture was stirred at 21 °C for 2 h. After the reaction

completion shown by TLC, water (6 mL) was added to the reaction

vial. The reaction mixture was extracted by dichloromethane (15 mL

× 3). The combined organic layer was dried by sodium sulfate and

concentrated. The residue was puriﬁed by ﬂash column chromatog-

raphy (hexanes/ethyl acetate = 12:1) to provide the desired

benzylation product 5.

POCl₃(1 mmol, 90 μL) was added dropwise to DMF (2 mL) at 0

°C under argon. After stirring for 10 min, a solution of compound 2

(1 mmol, 334 mg) in DMF (2 mL) was added slowly with stirring.

The mixture was kept at 21 °C overnight. The reaction was quenched

by adding water (5 mL) at 0 °C and then extracted with

dichloromethane (10 mL × 3). The combined organic layer was

dried by sodium sulfate and concentrated. The residue was puriﬁed by

ﬂash column chromatography (hexanes/ethyl acetate = 4:1) to

provide the desired aldehyde 12.

reaction was complete as shown by TLC and it was then quenched by

adding sat. ammonium chloride (10 mL). The resulting mixture was

extracted with dichloromethane (20 mL × 3), and the organic phases

were combined and evaporated on a rotary evaporator. The residue

was puriﬁed by ﬂash column chromatography (hexanes/ethyl acetate

= 10:1) to aﬀord the desired product 2-(3,5-bis(triﬂuoromethyl)-

phenyl)ethan-1-ol (yield: 90%, 464 mg).

To a solution of 2-(3,5-bis(triﬂuoromethyl)phenyl)ethan-1-ol (0.2

mmol, 51.6 mg) in dichloromethane (2 mL) were added triethyl-

amine (0.22 mmol, 31 μL) and mesyl chloride (MsCl, 0.2 mmol, 17

μL) at 0 °C. After stirring for 1 h, the reaction was complete as shown

by TLC. The solvent was removed by ﬂowing air over the open ﬂask.

The residue was puriﬁed by ﬂash column chromatography (hexanes/

ethyl acetate = 10:1) to aﬀord the desired product 3,5-bis-

(triﬂuoromethyl)phenethyl methanesulfonate (yield: 82%, 55.1 mg).

A ﬂask containing NaH (60%, 0.22 mmol, 8.8 mg) and a stir bar

was sealed, vacuumed, and reﬁlled with argon three times. Anhydrous

THF (3 mL) and a solution of tert-butyl (E)-2-cyano-3-(1H-indol-3-

yl)acrylate (0.2 mmol, 53.6 mg) in THF (2 mL) were added into the

reaction ﬂask. The reaction mixture was stirred for 30 min, and then

3,5-bis(triﬂuoromethyl)phenethyl methanesulfonate (0.164 mmol,

55.1 mg) in 2 mL of THF was added. The reaction was stirred for

24 h and quenched by sat. NH₄Cl solution. The resulting mixture was

extracted with dichloromethane (4 mL × 3), and the organic phases

were combined and evaporated on a rotary evaporator. The residue

was puriﬁed by ﬂash column chromatography (hexanes/ethyl acetate

= 10:1) to aﬀord the desired product tert-butyl (E)-3-(1-(3,5-

bis(triﬂuoromethyl)-phenethyl)-1H-indol-3-yl)-2-cyano-acrylate

(yield: 68%, 69 mg).

To the solution of aldehyde 3 (1 mmol, 372 mg) in ethanol (2 mL)

were added ethyl 2-cyanoacetate (1.3 equiv, 1.3 mmol, 140 μL) and ^L-

proline (40 mol %, 0.4 mmol, 58 mg). The reaction was stirred at 21

°C for 12 h, and a yellow solid precipitated gradually. After

completion of the reaction, ice-cold water (2 mL) was added into

the reaction vial. The solid was separated by Buchner funnel ﬁltration

and washed with water (2 mL × 3) and dried to aﬀord the desired

product JXL082.

To the solution of JXL082 (0.21 mmol, 100 mg) in THF (2 mL)

was added 0.5 N LiOH solution (3 equiv, 0.4 mmol, 0.8 mL). The

reaction mixture was stirred at 21 °C for 1 h. After the reaction

completion shown by TLC, THF was evaporated. Concentrated HCl

was added dropwise to acidify the reaction mixture until pH was

lower than 1; meanwhile, a yellow solid precipitated. Ice-cold water (5

mL) was added to the reaction mixture, and the solid was ﬁltrated by

a PYREX Hirsch-Type Funnel with a fritted disc and washed with

water (5 mL × 3). After drying by vacuum, the solid was washed with

2 mL of solvent mixture (hexanes/ethyl acetate = 5:1) 5−10 times

and monitored by TLC until nonpolar impurities disappeared (the

nonpolar compound was the retro-Aldol condensation product, which

can be recovered from the ﬁltrate). Finally, the purity of the product

JXL069 was checked by NMR.

To a solution of methyl tert-butyl (E)-3-(1-(3,5-bis-

(triﬂuoromethyl)phenethyl)-1H-indol-3-yl)-2-cyanoacrylate (0.1

mmol, 50.8 mg) in dichloromethane (2 mL) was added triﬂuoroacetic

acid (3 equiv, 0.3 mmol, 34 μL). The reaction mixture was stirred at

21 °C for 30 min, and a yellow solid precipitated. After the reaction

was complete as shown by TLC, the reaction solvent was evaporated

by ﬂowing air over the open ﬂask. The solid was washed with 2 mL of

solvent mixture (hexanes/ethyl acetate = 5:1) 5−10 times and

monitored by TLC until all of the nonpolar impurities disappeared.

Finally, the purity of the product was checked by NMR. Yield: 87%,

39 mg.

Experimental Details for the Synthesis of JXL080. To a solution

of ethyl (E)-2-cyano-3-(1H-indol-3-yl)acrylate (0.5 mmol, 112 mg) in

dichloromethane (5 mL) were added 2-(3,5-bis(triﬂuoromethyl)-

phenyl)acetic acid (0.55 mmol, 150 mg), 4-(N,N-dimethylamino)-

pyridine (DMAP; catalytic amount, 6 mg), and DCC (0.5 mmol, 103

mg) at 0 °C. The mixture was allowed to reach 21 °C and stirred

overnight. The white precipitate was ﬁltered, and the resulting

solution was concentrated in vacuum. The solid was puriﬁed by ﬂash

column chromatography (hexanes/ethyl acetate = 10:1) to aﬀord the

desired product (yield: 78%, 192.6 mg).

Experimental Details for the Synthesis of JXL083. To a solution

of 1-(3,5-bis(triﬂuoromethyl)benzyl)-1H-indole-3-carbaldehyde (10

mmol, 3.71 g) in acetone (60 mL) were added 2-methyl-2-butene (9

mL), NaH₂PO₄(3 equiv, 4.4 g), and NaClO₂(6.6 mmol, 6 g) in 6

mL of water. The reaction mixture was stirred at 21 °C for 24 h. After

the reaction was complete as shown by TLC, the reaction solvent was

evaporated on a rotary evaporator. The crude material was dissolved

in ethyl acetate (30 mL) and water (30 mL) and extracted with ethyl

acetate (30 mL × 3). The organic phase was combined, dried with

sodium sulfate, and evaporated on a rotary evaporator. The solid was

puriﬁed by ﬂash column chromatography (hexanes/ethyl acetate =

2:1) to aﬀord the desired product 1-(3,5-bis(triﬂuoromethyl)benzyl)-

1H-indole-3-carboxylic acid (yield: 89%, 3.44 g).

A 100 mL round-bottom ﬂask with a stir bar containing the

carboxylic acid (5 mmol, 1935 mg) from the previous step was sealed,

vacuumed, and reﬁlled with argon three times. To the ﬂask was added

50 mL of dichloromethane and oxalyl chloride (25 mmol, 2.1 mL)

dropwise. The reaction mixture was stirred at 21 °C for 1.5 h. The

reaction solvent was evaporated by vacuum, and the resulting

compound was used for the next step.

The following compounds were synthesized by a route similar to

that described for JXL069: JXL072, JXL073, JXL076, JXL077,

JXL082, JXL087, and JXL088.

Experimental Details for the Synthesis of JXL079. A ﬂask

containing Mg powder (10 mmol, 240 mg) and a stir bar was sealed

and vacuumed and reﬁlled with argon three times. Anhydrous diethyl

ether (32 mL) and bis(triﬂuoromethyl)benzyl bromide (8 mmol, 1.46

mL) were added to the reaction ﬂask. The reaction mixture was

stirred and reﬂuxed for 30 min, and then ground dry ice powder (5 g)

was added into the reaction ﬂask. After 1 h, the reaction was complete

as shown by TLC. The extra Mg powder was ﬁltered oﬀ, and the

solvent was evaporated under vacuum. HCl (1 N, 20 mL) was added

to the residue, and the precipitate was ﬁltered and dried to provide

the desired carboxylic acid.

A ﬂask containing a stir bar was sealed, vacuumed, and reﬁlled with

argon three times. Anhydrous dichloromethane (20 mL) and DIBAL

(1 M in hexanes, 6 mmol, 6 mL) were added to the ﬂask. The crude

carboxylic acid (2 mmol, 544 mg) dissolved in dry dichloromethane

(10 mL) was added to the reaction ﬂask at −78 °C. After 2 h, the

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A 100 mL round-bottom ﬂask with a stir bar was sealed, vacuumed,

and reﬁlled with argon three times. Diisopropylamine (5.5 mmol, 765

μL) and THF (10 mL) were added into the ﬂask and it was cooled to

−78 °C. n-BuLi (2.5 M in hexanes, 5 mmol, 2 mL) was added slowly

to the ﬂask. After the mixture had stirred for 30 min, a solution of

ethyl 2-cyanoacetate (5 mmol, 590 μL) in THF (10 mL) was added

slowly to the ﬂask. After the mixture had stirred for 1 h, a solution of

acyl chloride (5 mmol from the previous step) in THF (5 mL) was

added slowly to the reaction mixture. After 1 h, the reaction was

quenched by adding an aqueous 1 M HCl solution (10 mL) and

extracted with ethyl acetate (10 mL × 3). The organic phase was

combined, dried with sodium sulfate, and evaporated by a rotary

evaporator. The solid was puriﬁed by ﬂash column chromatography

(hexanes/ethyl acetate = 10:1) to aﬀord the desired product ethyl

(Z)-3-(1-(3,5-bis(triﬂuoromethyl)benzyl)-1H-indol-3-yl)-2-cyano-3-

hydroxyacrylate (JXL083) (yield: 80%, 1.93 g).

Experimental Details for the Synthesis of JXL084. To a solution

of JXL083 (0.5 mmol, 240 mg) in dichloromethane (10 mL) were

added pyridine (0.5 mmol, 40 μL) and acetyl chloride (1.0 mmol, 84

μL). The reaction mixture was stirred for 1 h, and TLC indicated that

the reaction was complete. The reaction solvent was evaporated by

ﬂowing air over the open ﬂask. The residue was puriﬁed by ﬂash

column chromatography (hexanes/ethyl acetate = 10:1) to aﬀord the

desired product ethyl (Z)-3-acetoxy-3-(1-(3,5-bis(triﬂuoromethyl)-

benzyl)-1H-indol-3-yl)-2-cyanoacrylate (JXL084) (yield: 86%, 225

mg).

Experimental Details for the Synthesis of JXL085. To a solution

of JXL083 (0.5 mmol, 240 mg) in dichloromethane (10 mL) were

added triethylamine (1.0 mmol, 139.5 μL) and phosphoryl chloride

(0.55 mmol, 520 μL). The reaction mixture was stirred for 1 h at

reﬂux, and TLC indicated that the reaction was complete. The

reaction solvent was evaporated by ﬂowing air over the open ﬂask.

The residue was puriﬁed by ﬂash column chromatography (hexanes/

ethyl acetate = 10:1) to aﬀord the desired product ethyl (Z)-3-(1-

(3,5-bis(triﬂuoromethyl)benzyl)-1H-indol-3-yl)-3-chloro-2-cyanoa-

crylate (JXL085) (yield: 84%, 210 mg).

Experimental Details for the Synthesis of JXL086. To the

solution of 1-(3,5-bis(triﬂuoromethyl)benzyl)-1H-indole-3-carboxal-

dehyde (1 mmol, 371 mg) in ethanol (3 mL) were added diethyl

cyanomethylphosphate (1.3 equiv, 1.3 mmol, 204 μL) and ^L-proline

(40 mol%, 0.4 mmol, 58 mg). The reaction was stirred at 50 °C for 24

h. After completion of the reaction as indicated by TLC, the reaction

solvent was evaporated by ﬂowing air over the open ﬂask. The solid

was puriﬁed by ﬂash column chromatography (hexanes/ethyl acetate

= 2:1) to aﬀord the desired product JXL086 (yield: 90%, 477 mg).

Experimental Details for the Synthesis of JXL096. A solution of

JXL086 (30 mg, 0.057 mmol) in dichloromethane (2 mL) was cooled

to 0 °C and bromotrimethylsilane (40 μL, 0.3 mmol) was added

dropwise under argon. The mixture was warmed to 21 °C and stirred

for 12 h. The solvent was evaporated under vacuum, and the resulting

residue was then dissolved in methanol (2 mL). The mixture was

stirred at 21 °C for 2 h. Evaporation of all volatiles under vacuum gave

the phosphoric acid JXL096 (yield: 92%, 25 mg).

= 7.0 Hz, 3H). ¹³C NMR (126 MHz, CDCl₃) δ 163.6, 147.0, 136.6,

133.0, 127.3, 124.0, 122.5, 118.9, 118.4, 113.3, 110.3, 92.6, 61.8, 14.5.

HRMS (ESI, m/z) calcd for C₁₄H₁₂N₂O₂([M − H]⁻): 239.0820.

Found: 239.0818.

Ethyl (E)-2-Cyano-3-(1-phenyl-1H-indol-3-yl)acrylate (JXL004).

¹H NMR (500 MHz, CDCl₃) δ 8.71 (s, 1H), 8.66 (s, 1H), 7.90 (d, J

= 7.2 Hz, 1H), 7.54 (m, 6H), 7.36 (m, 2H), 4.39 (q, J = 7.1 Hz, 2H),

1.42 (t, J = 7.1 Hz, 3H). ¹³C NMR (126 MHz, CDCl₃) δ 163.6, 145.6,

137.8, 136.4, 133.2, 129.9, 128.5, 124.8, 124.4, 123.0, 118.5, 117.9,

111.6, 111.5, 95.4, 62.0, 14.3. HRMS (ESI, m/z) calcd for

C₂₀H₁₆N₂O₂([M + H]⁺): 317.1290. Found: 317.1269.

Ethyl (E)-3-(6-Chloro-1-phenyl-1H-indol-3-yl)-2-cyanoacrylate

(JXL005). ¹H NMR (500 MHz, CDCl₃) δ 8.67 (s, 1H), 8.58 (s,

1H), 7.81 (d, J = 8.5 Hz, 1H), 7.60 (m, 2H), 7.52 (m, 4H), 7.34 (d, J

= 8.4 Hz, 1H), 4.39 (q, J = 7.1 Hz, 2H), 1.41 (t, J = 7.1 Hz, 3H). ¹³C

NMR (126 MHz, CDCl₃) δ 163.4, 145.1, 137.3, 136.8, 133.5, 130.5,

130.0, 128.8, 126.8, 124.8, 123.6, 119.5, 117.5, 111.6, 111.4, 96.4,

62.1, 14.2. HRMS (ESI, m/z) calcd for C₂₀H₁₅ClN₂O₂([M + H]⁺):

351.0900. Found: 351.0907.

Ethyl (E)-2-Cyano-3-(1-(2-methoxyphenyl)-1H-indol-3-yl)-

1

acrylate (JXL006). H NMR (500 MHz, CDCl₃) δ 8.67 (s, 1H),

8.66 (s, 1H), 7.89 (d, J = 7.8 Hz, 1H), 7.49 (app. t, J = 8.6 Hz, 1H),

7.41 (d, J = 7.6 Hz, 1H), 7.35 (app. t, J = 7.3 Hz, 1H), 7.30 (t, J = 7.5

Hz, 1H), 7.23 (d, J = 8.1 Hz, 1H), 7.13 (m, 2H), 4.39 (q, J = 7.1 Hz,

2H), 3.81 (s, 3H), 1.41 (t, J = 7.1 Hz, 3H). ¹³C NMR (126 MHz,

CDCl₃) δ 163.8, 154.2, 146.0, 137.2, 135.2, 130.3, 127.8, 126.0, 124.0,

122.6, 120.8, 118.2, 118.0, 112.3, 111.8, 111.0, 94.7, 61.8, 55.7, 14.3.

HRMS (ESI, m/z) calcd for C₂₁H₁₈N₂O₃([M + H]⁺): 347.1396.

Found: 347.1403.

Ethyl (E)-2-Cyano-3-(1-(4-methoxyphenyl)-1H-indol-3-yl)-

1

acrylate (JXL007). H NMR (500 MHz, CDCl₃) δ 8.65 (s, 1H),

8.64 (s, 1H), 7.89 (d, J = 7.2 Hz, 1H), 7.44 (m, 3H), 7.35 (m, 2H),

7.07 (d, J = 8.8 Hz, 2H), 4.38 (q, J = 7.1 Hz, 2H), 3.90 (s, 3H), 1.41

(t, J = 7.1 Hz, 3H). ¹³C NMR (126 MHz, CDCl₃) δ 163.8, 159.5,

145.6, 136.8, 133.5, 130.6, 128.3, 126.2, 124.2, 122.9, 118.4, 118.0,

115.0, 111.5, 111.1, 94.9, 61.9, 55.6, 14.2. HRMS (ESI, m/z) calcd for

C₂₁H₁₈N₂O₃([M + H]⁺): 347.1396. Found: 347.1401.

Ethyl (E)-2-Cyano-3-(1-(4-ﬂuorobenzyl)-1H-indol-3-yl)acrylate

1

(JXL008). H NMR (500 MHz, CDCl₃) δ 8.60 (app. s, 2H), 7.85

(d, J = 6.8 Hz, 1H), 7.32 (m, 3H), 7.15 (m, 2H), 7.03 (app. t, 2H),

5.39 (s, 2H), 4.37 (q, J = 7.1 Hz, 2H), 1.40 (t, J = 7.1 Hz, 3H). ¹³C

NMR (126 MHz, CDCl₃) δ 163.7, 162.5 (d, J_c−f= 247.7 Hz), 145.7,

136.1, 133.8, 130.9, 128.6, 128.5, 124.0, 122.7, 118.6, 118.0, 116.0 (d,

J

_c−f= 21.9 Hz), 110.9, 110.4, 94.6, 61.9, 50.7, 14.2. HRMS (ESI, m/z)

calcd for C₂₁H₁₇FN₂O₂([M + H]⁺): 347.1352. Found: 347.1351.

Ethyl (E)-2-Cyano-3-(1-(3,4-diﬂuorobenzyl)-1H-indol-3-yl)-

1

acrylate (JXL009). H NMR (500 MHz, CDCl₃) δ 8.60 (s, 1H),

8.59 (s, 1H), 7.86 (d, J = 7.8 Hz, 1H), 7.33 (m, 2H), 7.28 (s, 1H),

7.13 (m, 1H), 6.95 (m, 1H), 6.89 (m, 1H), 5.39 (s, 2H), 4.38 (q, J =

7.1 Hz, 2H), 1.40 (t, J = 7.1 Hz, 3H). ¹³C NMR (126 MHz, CDCl₃) δ

163.7, 150.7 (dd, J = 251.2, 13.2 Hz), 150.2 (dd, J = 250.4, 12.6 Hz),

145.7, 136.1, 133.7, 132.3, 128.6, 124.3, 122.9, 122.7, 120.0, 118.8,

118.0 (d, J = 17.5 Hz), 115.9 (d, J = 18.0 Hz), 110.8, 110.6, 95.2,

62.1, 50.4, 14.4. HRMS (ESI, m/z) calcd for C₂₁H₁₆F₂N₂O₂([M +

H]⁺): 367.1258. Found: 367.1272.

Characterization Data for JXL001−JX096. (E)-2-Cyano-3-(1-

phenyl-1H-indol-3-yl)acrylic Acid (JXL001/UK-5099). ¹H NMR (500

MHz, DMSO-d₆) δ 8.59 (s, 1H), 8.56 (s, 1H), 8.06 (m, 1H), 7.65 (m,

4H), 7.53 (m, 2H), 7.34 (m, 2H). ¹³C NMR (126 MHz, DMSO-d₆) δ

164.5, 145.6, 137.7, 136.3, 133.6, 130.5, 128.9, 128.0, 125.0, 124.9,

123.3, 119.9, 118.4, 111.9, 96.7. HRMS (ESI, m/z) calcd for

C₁₈H₁₂N₂O₂([M − H]⁻): 287.0821. Found: 287.0826.

Ethyl (E)-2-Cyano-3-(1-(3,5-diﬂuorobenzyl)-1H-indol-3-yl)-

1

acrylate (JXL010). H NMR (500 MHz, CDCl₃) δ 8.61 (s, 1H),

8.59 (s, 1H), 7.87 (d, J = 7.1 Hz, 1H), 7.33 (m, 2H), 7.26 (d, J = 7.2

Hz, 1H), 6.75 (app. t, J = 8.7 Hz, 1H), 6.64 (app. d, J = 5.7 Hz, 2H),

5.41 (s, 2H), 4.38 (q, J = 7.1 Hz, 2H), 1.40 (t, J = 7.1 Hz, 3H). ¹³C

NMR (126 MHz, CDCl₃) δ 163.6, 163.3 (dd, J_c−f= 251.0, 12.5 Hz),

145.5, 139.2, 136.0, 133.6, 128.4, 124.3, 122.9, 118.7, 118.0, 110.7,

110.6, 109.5 (dd, J_c−f= 19.9, 6.4 Hz), 103.8 (t, J_c−f= 25.2 Hz), 95.3,

62.0, 50.4, 14.2. HRMS (ESI, m/z) calcd for C₂₁H₁₆F₂N₂O₂([M +

H]⁺): 367.1258. Found: 367.1264.

(E)-2-Cyano-3-(1H-indol-3-yl)acrylic Acid (JXL002). ¹H NMR

(500 MHz, DMSO-d₆) δ 12.48 (s, 1H), 8.51 (s, 1H), 8.49 (s, 1H),

7.91 (d, J = 6.5 Hz, 1H), 7.53 (d, J = 7.0 Hz, 1H), 7.23 (m, 2H). ¹³C

NMR (126 MHz, DMSO-d₆) δ 165.0, 146.5, 136.5, 132.4, 127.3,

123.9, 122.4, 118.9, 118.8, 113.2, 110.2, 94.0. HRMS (ESI, m/z) calcd

for C₁₂H₈N₂O₂([M − H]⁻): 211.0508. Found: 211.0511.

Ethyl (E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-indol-3-yl)-2-

1

cyanoacrylate (JXL011). H NMR (500 MHz, CDCl₃) δ 8.62 (s,

Ethyl (E)-2-Cyano-3-(1H-indol-3-yl)acrylate (JXL003). H NMR

(500 MHz, CDCl₃) δ 12.55 (s, 1H), 8.53 (s, 1H), 8.52 (s, 1H), 7.92

(d, J = 7.6 Hz, 1H), 7.53 (d, J = 7.8 Hz, 1H), 7.26 (app. t, J = 7.4 Hz,

1H), 7.22 (app. t, J = 7.4 Hz, 1H), 4.24 (q, J = 7.0 Hz, 2H), 1.26 (t, J

1H), 8.61 (s, 1H), 7.90 (d, J = 7.7 Hz, 1H), 7.85 (s, 1H), 7.57 (s,

2H), 7.35 (m, 2H), 7.23 (d, J = 7.8 Hz, 1H), 5.56 (s, 2H), 4.38 (q, J =

7.1 Hz, 2H), 1.40 (t, J = 7.1 Hz, 3H). ¹³C NMR (126 MHz, CDCl₃) δ

2056

https://dx.doi.org/10.1021/acs.jmedchem.0c01570

J. Med. Chem. 2021, 64, 2046−2063

Journal of Medicinal Chemistry

pubs.acs.org/jmc

Article

163.6, 145.6, 138.1, 136.0, 133.4, 132.7 (q, J = 33.8 Hz), 128.6, 126.8,

124.7, 123.2, 122.9 (q, J = 273.4 Hz), 122.6, 119.0, 118.0, 111.2,

110.4, 95.9, 62.1, 50.5, 14.3. HRMS (ESI, m/z) calcd for

C₂₃H₁₆F₆N₂O₂([M + H]⁺): 467.1194. Found: 467.1206.

7.28 (m, 2H), 5.83 (s, 2H). ¹³C NMR (126 MHz, DMSO-d₆) δ 164.7,

145.7, 140.3, 136.3, 134.8, 131.1, 130.8 (q, J = 31.1 Hz), 128.9, 128.7,

127.9, 124.8, 124.3, 122.9 (q, J = 273.4 Hz), 122.2, 119.3, 118.3, 95.6,

49.2. HRMS (ESI, m/z) calcd for C₂₁H₁₂F₆N₂O₂([M + H]⁺):

439.0881. Found: 439.0880.

(E)-2-Cyano-3-(1-(4-methoxyphenyl)-1H-indol-3-yl)acrylic Acid

1

(JXL012). H NMR (500 MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.52 (s,

2-((1-Phenyl-1H-indol-3-yl)methylene)malononitrile (JXL021).

¹H NMR (500 MHz, CDCl₃) δ 8.62 (s, 1H), 8.13 (s, 1H), 7.80

(d, J = 7.4 Hz, 1H), 7.61 (m, 2H), 7.53 (m, 4H), 7.40 (m, 2H). ¹³C

NMR (126 MHz, CDCl₃) δ 149.8, 137.3, 136.5, 133.6, 130.1, 128.9,

127.7, 124.9, 124.8, 123.7, 118.2, 115.1, 115.0, 111.9, 111.8, 73.7.

HRMS (ESI, m/z) calcd for C₁₈H₁₁N₃([M + H]⁺): 321.0698. Found:

321.0695.

1H), 8.05 (d, J = 7.7 Hz, 1H), 7.58 (app. d, J = 8.7 Hz, 2H), 7.44 (m,

1H), 7.33 (m, 2H), 7.16 (app. d, J = 8.7 Hz, 2H), 3.82 (s, 3H). ¹³C

NMR (126 MHz, DMSO-d₆) δ 164.7, 160.5, 136.7, 133.9, 130.5,

127.8, 126.6, 124.8, 123.2, 120.0, 119.5, 118.6, 115.6, 115.4, 111.9,

110.9, 55.9. HRMS (ESI, m/z) calcd for C₁₉H₁₄N₂O₃([M + H]⁺):

319.1083. Found: 319.1077.

Ethyl (E)-2-Cyano-3-(pyridin-4-yl)acrylate (JXL022). ¹H NMR

(500 MHz, CDCl₃) δ 8.81 (d, J = 5.2 Hz, 2H), 8.18 (s, 1H), 7.74 (d, J

= 5.2 Hz, 2H), 4.41 (q, J = 7.1 Hz, 2H), 1.41 (t, J = 7.1 Hz, 3H). ¹³C

NMR (126 MHz, CDCl₃) δ 161.2, 152.0, 151.0, 137.9, 123.2, 114.2,

108.2, 63.2, 14.0. HRMS (ESI, m/z) calcd for C₁₁H₁₀N₂O₂([M +

H]⁺): 203.0820. Found: 203.0815.

(E)-3-(6-Chloro-1-phenyl-1H-indol-3-yl)-2-cyanoacrylic Acid

(JXL013). ¹H NMR (500 MHz, DMSO-d₆) δ 13.58 (br. s, 1H),

8.59 (s, 1H), 8.54 (s, 1H), 8.11 (d, J = 7.5 Hz, 1H), 7.67 (m, 4H),

7.53 (m, 2H), 7.35 (d, J = 7.5 Hz, 1H). ¹³C NMR (126 MHz, DMSO-

d₆) δ 164.3, 145.4, 137.3, 136.7, 134.4, 130.6, 129.6, 129.1, 126.7,

125.1, 123.5, 121.5, 118.2, 111.6, 111.3, 97.9. HRMS (ESI, m/z) calcd

for C₁₈H₁₁ClN₂O₂([M + H]⁺): 323.0587. Found: 323.0587.

(E)-2-Cyano-3-(1-(2-methoxyphenyl)-1H-indol-3-yl)acrylic Acid

(Z)-2-Imino-5-((1-phenyl-1H-indol-3-yl)methylene)thiazolidin-4-

1

one (JXL023). H NMR (500 MHz, DMSO-d₆) δ 11.94 (br. s, 1H),

1

9.33 (br. s, 1H), 8.97 (s, 1H), 7.64 (m, 10H). ¹³C NMR (126 MHz,

DMSO-d₆) δ 180.9, 174.8, 172.5, 138.6, 136.2, 130.5, 129.3, 128.1,

126.3, 124.8, 124.5, 122.2, 120.3, 119.7, 113.5, 111.5. HRMS (ESI, m/

z) calcd for C₁₈H₁₃N₃OS ([M + H]⁺): 320.0858. Found: 320.0862.

(Z)-5-((1-Phenyl-1H-indol-3-yl)methylene)thiazolidine-2,4-dione

(JXL014). H NMR (500 MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.47 (s,

1H), 8.02 (d, J = 7.4 Hz, 1H), 7.54 (m, 2H), 7.31 (m, 3H), 7.15 (m,

2H), 3.74 (s, 3H). ¹³C NMR (126 MHz, DMSO-d₆) δ 164.6, 154.2,

145.8, 137.3, 135.1, 131.1, 128.2, 127.3, 125.7, 124.6, 123.0, 121.5,

119.3, 118.5, 113.6, 112.2, 110.8, 96.1, 56.2. HRMS (ESI, m/z) calcd

for C₁₉H₁₄N₂O₃([M + H]⁺): 319.1083. Found: 319.1084.

1

(JXL024). H NMR (500 MHz DMSO-d₆) δ 7.98 (m, 2H), 7.79 (s,

(E)-2-Cyano-3-(1-(4-ﬂuorobenzyl)-1H-indol-3-yl)acrylic Acid

1H), 7.66 (app. d, J = 7.7 Hz, 2H), 7.62 (app. t, J = 7.7 Hz, 2H), 7.54

(d, J = 8.0 Hz, 1H), 7.49 (t, J = 7.1 Hz, 1H), 7.30 (m, 2H). ¹³C NMR

(126 MHz, DMSO-d₆) δ 172.5, 169.5, 138.4, 136.2, 130.5, 129.9,

128.3, 128.2, 125.0, 124.6, 122.4, 121.5, 121.4, 119.6, 113.0, 111.5.

HRMS (ESI, m/z) calcd for C₁₈H₁₂N₂O₂S ([M + H]⁺): 321.0698.

Found: 321.0695.

1

(JXL015). H NMR (500 MHz, DMSO-d₆) δ 8.64 (s, 1H), 8.46 (s,

1H), 7.93 (d, J = 7.1 Hz, 1H), 7.61 (d, J = 7.3 Hz, 1H), 7.33 (m, 2H),

7.26 (m, 2H), 7.16 (m, 2H), 5.60 (s, 2H). ¹³C NMR (126 MHz,

DMSO-d₆) δ 164.8, 162.0 (d, J_c−f= 244.3 Hz), 145.6, 136.4, 134.6,

133.1, 130.0, 128.0, 124.1, 122.8, 119.2, 118.5, 116.0 (d, J_c−f= 21.7

Hz), 112.0, 109.8, 95.0, 49.6. HRMS (ESI, m/z) calcd for

C₁₉H₁₃FN₂O₂([M + H]⁺): 321.1039. Found: 321.1046.

(E)-2-Cyano-3-(1-(3,4-diﬂuorobenzyl)-1H-indol-3-yl)acrylic Acid

(JXL016). ¹H NMR (500 MHz, DMSO-d₆) δ 13.34 (br. s, 1H),

8.62 (s, 1H), 8.47 (s, 1H), 7.94 (d, J = 7.3 Hz, 1H), 7.61 (d, J = 7.6

Hz, 1H), 7.40 (m, 2H), 7.27 (m, 2H), 7.10 (br. s, 1H), 5.61 (s, 2H).

¹³C NMR (126 MHz, DMSO-d₆) δ 164.8, 149.7 (dd, J_c−f= 253.3,

13.6 Hz), 149.4 (dd, J_c−f= 246.3, 11.6 Hz), 145.7, 136.4, 134.7, 134.5,

128.0, 124.7 (dd, J_c−f= 5.9, 3.0 Hz), 124.2, 122.8, 119.2, 118.4, 118.3

(d, J_c−f= 17.0 Hz), 117.1 (d, J_c−f= 17.6 Hz), 112.0, 109.9, 95.0, 49.3.

HRMS (ESI, m/z) calcd for C₁₉H₁₂F₂N₂O₂([M + H]⁺): 339.0945.

Found: 339.0946.

2-(Ethoxycarbonyl)-3-(1-phenyl-1H-indol-3-yl)acrylic Acid (a

1

Mixture of E/Z Isomers, 1:1 Ratio) (JXL025). H NMR (500 MHz,

DMSO-d₆) δ 13.08 (br. s, 1H), 7.87 (m, 3H), 7.61 (m, 4H), 7.52 (m,

2H), 7.30 (m, 2H), 4.26 (m, 2H), 1.23 (m, 3H). ¹³C NMR (126

MHz, DMSO-d₆) δ 168.9, 167.8, 166.1, 164.9, 138.3, 136.1, 132.9,

132.1, 131.1, 130.9, 130.8, 128.5, 124.9, 124.4, 122.6, 122.2, 119.5,

111.6, 111.0, 61.7, 61.3, 14.7, 14.4. HRMS (ESI, m/z) calcd for

C₂₀H₁₇NO₄([M + H]⁺): 336.1236. Found: 336.1239.

(E)-2-Cyano-3-(4-ﬂuoro-1-phenyl-1H-indol-3-yl)acrylic Acid

1

(JXL026). H NMR (500 MHz, DMSO-d₆) δ 8.65 (s, 1H), 8.61 (s,

1H), 7.67 (m, 4H), 7.57 (m, 1H), 7.36 (m, 2H), 7.19 (m, 1H). ¹³C

NMR (126 MHz, DMSO-d₆) δ 164.4, 156.8 (d, J_c−f= 245.6 Hz),

146.4, 138.7, 137.6, 133.4, 130.7, 129.5, 125.9, 125.4, 118.2, 116.3,

116.2, 109.5 (d, J_c−f= 34.5 Hz), 109.2 (d, J_c−f= 23.2 Hz), 98.0.

HRMS (ESI, m/z) calcd for C₁₈H₁₁FN₂O₂([M + H]⁺): 307.0880.

Found: 307.0885.

(E)-2-Cyano-3-(1-(3,5-diﬂuorobenzyl)-1H-indol-3-yl)acrylic Acid

1

(JXL017). H NMR (500 MHz, DMSO-d₆) δ 8.67 (s, 1H), 8.48 (s,

1H), 7.95 (d, J = 4.8 Hz, 1H), 7.59 (d, J = 4.4 Hz, 1H), 7.27 (m, 2H),

7.15 (s, 1H), 6.98 (br. s, 2H), 5.65 (s, 2H). ¹³C NMR (126 MHz,

DMSO-d₆) δ 164.8, 162.9 (dd, J_c−f= 247.0, 12.8 Hz), 145.6, 141.4,

(E)-2-Cyano-3-(6-ﬂuoro-1-phenyl-1H-indol-3-yl)acrylic Acid

(JXL027). ¹H NMR (500 MHz, DMSO-d₆) δ 13.59 (br. s, 1H),

8.62 (s, 1H), 8.59 (s, 1H), 8.16 (m, 1H), 7.66 (m, 4H), 7.56 (m, 1H),

7.36 (d, J = 9.2 Hz, 1H), 7.25 (m, 1H). ¹³C NMR (126 MHz, DMSO-

d₆) δ 164.6, 160.8 (d, J_c−f= 240.0 Hz), 145.7, 137.6, 136.6, 134.4,

130.8, 129.1, 125.0, 124.7, 121.6, 118.4, 111.8 (d, J_c−f= 24.2 Hz),

111.5, 98.7 (d, J_c−f= 26.2 Hz), 97.7. HRMS (ESI, m/z) calcd for

C₁₈H₁₁FN₂O₂([M + H]⁺): 307.0883. Found: 307.0886.

(E)-2-Cyano-3-(7-ﬂuoro-1-phenyl-1H-indol-3-yl)acrylic Acid

(JXL028). ¹H NMR (500 MHz, DMSO-d₆) δ 13.62 (br. s, 1H),

8.56 (s, 1H), 8.47 (s, 1H), 7.89 (br. s, 1H), 7.61 (m, 5H), 7.30 (br. s,

1H), 7.17 (br. s, 1H). ¹³C NMR (126 MHz, DMSO-d₆) δ 164.4,

149.7 (d, J_c−f= 247.5 Hz), 145.5, 139.0, 135.5, 131.9, 129.9, 129.3,

126.2, 124.3, 124.0, 118.3, 115.9, 111.8, 110.9 (d, J_c−f= 17.4 Hz),

98.1. HRMS (ESI, m/z) calcd for C₁₈H₁₁FN₂O₂([M + H]⁺):

307.0883. Found: 307.0870.

136.4, 134.8, 128.0, 124.2, 122.9, 119.3, 118.4, 111.9, 111.0 (d, J_c−f

=

26.1 Hz), 110.8, 103.8 (t, J_c−f= 26.5 Hz), 95.5, 49.5. HRMS (ESI, m/

z) calcd for C₁₉H₁₂F₂N₂O₂([M + H]⁺): 339.0945. Found: 339.0944.

Ethyl (E)-3-(1-Benzyl-1H-indol-3-yl)-2-cyanoacrylate (JXL018).

¹H NMR (500 MHz, CDCl₃) δ 8.61 (s, 1H), 8.60 (s, 1H), 7.85

(d, J = 7.9 Hz, 1H), 7.33 (m, 6H), 7.17 (m, 2H), 5.42 (s, 2H), 4.37

(q, J = 7.1 Hz, 2H), 1.40 (t, J = 7.1 Hz, 3H). ¹³C NMR (126 MHz,

CDCl₃) δ 163.8, 145.7, 136.2, 135.1, 134.0, 129.0, 128.5, 128.2, 126.8,

124.0, 122.6, 118.5, 118.1, 111.0, 110.3, 94.3, 61.8, 51.4, 14.2. HRMS

(ESI, m/z) calcd for C₂₁H₁₈N₂O₂([M + H]⁺): 331.1447. Found:

331.1442.

1

(E)-3-(1-Benzyl-1H-indol-3-yl)-2-cyanoacrylic Acid (JXL019). H

NMR (500 MHz, DMSO-d₆) δ 13.34 (br. s, 1H), 8.65 (s, 1H), 8.48

(s, 1H), 7.93 (d, J = 6.9 Hz, 1H), 7.60 (d, J = 6.8 Hz, 1H), 7.25 (m,

7H), 5.62 (s, 2H). ¹³C NMR (126 MHz, DMSO-d₆) δ 164.8, 145.7,

136.8, 136.5, 134.7, 129.1, 128.2, 127.6, 124.0, 122.7, 120.0, 119.2,

118.5, 112.1, 109.7, 94.7, 50.4. HRMS (ESI, m/z) calcd for

C₁₉H₁₄N₂O₂([M + H]⁺): 303.1133. Found: 303.1118.

(E)-2-Cyano-3-(5-ﬂuoro-1-phenyl-1H-indol-3-yl)acrylic Acid

(JXL029). ¹H NMR (500 MHz, DMSO-d₆) δ 13.57 (br. s, 1H),

8.67 (s, 1H), 8.60 (s, 1H), 8.01 (d, J = 9.0 Hz, 1H), 7.70 (m, 4H),

7.58 (m, 2H), 7.24 (m, 1H). ¹³C NMR (126 MHz, DMSO-d₆) δ

164.6, 159.4 (d, J_c−f= 237.8 Hz), 145.8, 137.7, 135.1, 133.1, 130.7,

129.2, 129.1, 125.2, 118.5, 113.7, 113.2 (d, J_c−f= 26.5 Hz), 111.4,

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-indol-3-yl)-2-cya-

1

noacrylic Acid (JXL020). H NMR (500 MHz, DMSO-d₆) δ 13.37

(br. s, 1H), 8.75 (s, 1H), 8.48 (s, 1H), 7.99 (m, 4H), 7.65 (s, 1H),

2057

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Article

105.7 (d, J_c−f= 24.2 Hz), 97.2. HRMS (ESI, m/z) calcd for

C₁₈H₁₁FN₂O₂([M + H]⁺): 307.0883. Found: 307.0872.

136.8, 135.2, 124.6, 121.1, 118.4, 111.3, 111.1, 110.3, 104.0 (t, J_c−f=

25.2 Hz), 98.7 (d, J_c−f= 26.5 Hz), 96.9, 49.6. HRMS (ESI, m/z) calcd

for C₁₉H₁₁F₃N₂O₂([M + H]⁺): 357.0851. Found: 357.0847.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-6-ﬂuoro-1H-indol-3-yl)-

(E)-2-Cyano-3-(2-ﬂuorophenyl)acrylic Acid (JXL030). ¹H NMR

(500 MHz, DMSO-d₆) δ 8.49 (s, 1H), 8.31 (app. t, J = 7.2 Hz, 1H),

7.63 (m, 1H), 7.36 (app. t, J = 7.4 Hz, 1H), 7.30 (m, 1H). ¹³C NMR

(126 MHz, DMSO-d₆) δ 162.9, 161.5 (d, J_c−f= 256.2 Hz), 145.4 (d,

1

2-cyanoacrylic Acid (JXL041). H NMR (500 MHz, DMSO-d₆) δ

8.74 (s, 1H), 8.48 (s, 1H), 8.06 (app. s, 3H), 8.01 (dd, J = 8.7, 5.1 Hz,

1H), 7.66 (dd, J = 9.8, 2.0 Hz, 1H), 7.13 (dt, J = 9.3, 2.1 Hz, 1H),

5.78 (s, 2H). ¹³C NMR (126 MHz, DMSO-d₆) δ 164.7, 159.9 (d, J_c−f

= 264.6 Hz), 145.8, 140.2, 136.8, 135.3, 131.1 (q, J_c−f= 33.3 Hz),

129.1, 124.5, 123.6 (q, J_c−f= 273.7 Hz), 122.5, 121.2, 118.4, 111.4 (d,

J_c−f= 25.2 Hz), 110.4, 98.6 (d, J_c−f= 27.2 Hz), 96.7, 49.4. HRMS

(ESI, m/z) calcd for C₂₁H₁₁F₇N₂O₂([M + H]⁺): 457.0787. Found:

457.0783.

J_c−f= 7.8 Hz), 135.0 (d, J_c−f= 9.2 Hz), 128.7, 124.7, 119.8 (d, J_c−f

=

10.9 Hz), 115.8 (d, J_c−f= 21.9 Hz), 114.9, 105.9.HRMS (ESI, m/z)

calcd for C₁₀H₆FNO₂([M − H]⁻): 190.0304. Found: 190.0305.

(E)-2-Cyano-3-(4-ﬂuorophenyl)acrylic Acid (JXL032). ¹H NMR

(500 MHz, DMSO-d₆) δ 8.30 (s, 1H), 8.10 (m, 2H), 7.29 (m, 2H).

¹³C NMR (126 MHz, DMSO-d₆) δ 165.2 (d, J_c−f= 255.2 Hz), 163.5,

153.1, 133.3 (d, J_c−f= 9.3 Hz), 128.3, 116.0 (d, J_c−f= 22.4 Hz), 115.3,

103.2. HRMS (ESI, m/z) calcd for C₁₀H₆FNO₂([M − H]⁻):

190.0304. Found: 190.0303.

(E)-2-Cyano-3-(3-ﬂuoro-4-methylphenyl)acrylic Acid (JXL042).

¹H NMR (500 MHz, DMSO-d₆) δ 8.28 (s, 1H), 7.78 (m, 2H),

7.49 (t, J = 8.0 Hz, 1H), 2.30 (s, 3H). ¹³C NMR (126 MHz, DMSO-

(E)-2-Cyano-3-(4-(triﬂuoromethyl)phenyl)acrylic Acid (JXL034).

¹H NMR (500 MHz, DMSO-d₆) δ 8.39 (s, 1H), 8.17 (d, J = 7.7 Hz,

2H), 7.84 (d, J = 7.7 Hz, 2H). ¹³C NMR (126 MHz, DMSO-d₆) δ

162.8, 152.6, 135.4, 133.1 (q, J_c−f= 32.9 Hz), 131.0, 125.7, 123.7 (q,

J_c−f= 272.2 Hz), 114.8, 106.7. HRMS (ESI, m/z) calcd for

C₁₁H₆F₃NO₂([M − H]⁻): 240.0272. Found: 240.0271.

d₆) δ 163.6, 160.9 (d, J_c−f= 244.6 Hz), 153.5, 133.0, 131.6 (d, J_c−f

=

7.5 Hz), 130.8 (d, J_c−f= 17.6 Hz), 127.2, 117.9 (d, J_c−f= 23.9 Hz),

116.4, 104.5, 15.0. HRMS (ESI, m/z) calcd for C₁₁H₈FNO₂([M −

H]⁻): 204.0461. Found: 204.0460.

(E)-2-Cyano-3-(3,4-diﬂuorophenyl)acrylic Acid (JXL043). ¹H

NMR (500 MHz, DMSO-d₆) δ 8.32 (s, 1H), 8.09 (d, J = 8.0 Hz,

1H), 7.94 (br. s, 1H), 7.67 (d, J = 8.8 Hz, 1H). ¹³C NMR (126 MHz,

DMSO-d₆) δ 163.3, 152.3, 152.2 (dd, J_c−f= 255.4, 12.6 Hz), 149.9

(dd, J_c−f= 248.2, 12.6 Hz), 129.8, 128.8, 120.1 (d, J_c−f= 17.6 Hz),

119.1 (d, J_c−f= 17.6 Hz), 116.3, 105.8. HRMS (ESI, m/z) calcd for

C₁₀H₅F₂NO₂([M − H]⁻): 208.0210. Found: 208.0212.

(E)-3-(4-Chloro-1-phenyl-1H-indol-3-yl)-2-cyanoacrylic Acid

1

(JXL035). H NMR (500 MHz, DMSO-d₆) δ 9.22 (s, 1H), 8.68 (s,

1H), 7.64 (m, 3H), 7.59 (m, 2H), 7.48 (d, J = 7.6 Hz, 1H), 7.38 (m,

1H), 7.32 (m, 1H). ¹³C NMR (126 MHz, DMSO-d₆) δ 164.5, 146.4,

138.0, 137.4, 134.5, 130.7, 130.5, 129.5, 125.7, 125.3, 124.9, 123.7,

118.4, 111.7, 110.9, 97.5. HRMS (ESI, m/z) calcd for C₁₈H₁₁ClN₂O₂

([M + H]⁺): 323.0587. Found: 323.0587.

(E)-2-Cyano-3-(2,4-diﬂuorophenyl)acrylic Acid (JXL044). ¹H

NMR (500 MHz, DMSO-d₆) δ 14.2 (br. s, 1H), 8.27 (s, 1H), 8.24

(m, 1H), 7.52 (m, 1H), 7.35 (dt, J = 8.6, 2.2 Hz, 1H). ¹³C NMR (126

MHz, DMSO-d₆) δ 165.5 (dd, J_c−f= 255.8, 12.6 Hz), 163.1, 161.9

(E)-2-Cyano-3-(1-(3,4-diﬂuorobenzyl)-5-ﬂuoro-1H-indol-3-yl)-

1

acrylic Acid (JXL036). H NMR (500 MHz, DMSO-d₆) δ 8.69 (s,

1H), 8.47 (s, 1H), 7.84 (d, J = 9.6 Hz, 1H), 7.63 (dd, J = 8.9, 4.3 Hz,

1H), 7.41 (m, 2H), 7.14 (m, 2H), 5.61 (s, 2H). ¹³C NMR (126 MHz,

(dd, J_c−f= 270.5, 12.6 Hz), 145.1, 131.1, 117.0, 115.9, 113.5 (d, J_c−f

=

DMSO-d₆) δ 164.9, 159.4 (d, J_c−f= 237.3 Hz), 149.8 (dd, J_c−f

=

22.7 Hz), 106.8, 105.6 (t, J_c−f= 26.5 Hz). HRMS (ESI, m/z) calcd for

C₁₀H₅F₂NO₂([M − H]⁻): 208.0210. Found: 208.0208.

247.1, 12.7 Hz), 149.5 (dd, J_c−f= 246.6, 12.3 Hz), 146.0, 136.1, 134.5,

133.1, 129.1, 125.0, 118.5, 118.4, 117.4, 113.6, 112.5 (d, J_c−f= 26.2

Hz), 110.1, 105.2 (d, J_c−f= 25.1 Hz), 95.5, 49.7. HRMS (ESI, m/z)

calcd for C₁₉H₁₁F₃N₂O₂([M + H]⁺): 357.0851. Found: 357.0857.

(E)-2-Cyano-3-(1-(3,5-diﬂuorobenzyl)-5-ﬂuoro-1H-indol-3-yl)-

(E)-3-(3,5-Bis(triﬂuoromethyl)phenyl)-2-cyanoacrylic Acid

1

(JXL045). H NMR (500 MHz, CD3OD) δ 8.59 (s, 2H), 8.48 (s,

1H), 8.19 (s, 1H). HRMS (ESI, m/z) calcd for C₁₂H₅F₆NO₂([M +

H]⁺): 308.0146. Found: 308.0143.

1

acrylic Acid (JXL037). H NMR (500 MHz, DMSO-d₆) δ 8.71 (s,

(E)-2-Cyano-3-(4-ﬂuoro-3-(triﬂuoromethyl)phenyl)acrylic Acid

1

(JXL047). H NMR (500 MHz, DMSO-d₆) δ 8.46 (dd, J = 7.1, 1.8

1H), 8.48 (s, 1H), 7.85 (dd, J = 9.6, 2.0 Hz, 1H), 7.62 (dd, J = 8.9, 4.3

Hz, 1H), 7.16 (m, 2H), 7.00 (app. d, J = 6.2 Hz, 2H), 5.65 (s, 2H).

¹³C NMR (126 MHz, DMSO-d₆) δ 164.9, 163.0 (d, J_c−f= 247.7 Hz),

159.3 (d, J_c−f= 237.6 Hz), 145.9, 141.4, 136.3, 133.2, 129.1, 118.5,

113.6, 112.6 (d, J_c−f= 26.3 Hz), 111.3, 110.2, 105.3 (d, J_c−f= 24.9

Hz), 104.0 (t, J_c−f= 25.2 Hz), 95.9, 49.8. HRMS (ESI, m/z) calcd for

C₁₉H₁₁F₃N₂O₂([M + H]⁺): 357.0851. Found: 357.0851.

Hz, 1H), 8.44 (s, 1H), 8.40 (m, 1H), 7.75 (m, 1H). ¹³C NMR (126

MHz, DMSO-d₆) δ 163.3, 161.1 (d, J_c−f= 262.1 Hz), 152.2, 137.6 (d,

J

_c−f= 10.1 Hz), 130.5, 129.3, 122.6 (q, J_c−f= 272.8 Hz), 118.9 (d, J_c−f

= 21.2 Hz), 118.2 (qd, J_c−f= 32.9, 12.6 Hz), 116.2, 106.3. HRMS

(ESI, m/z) calcd for C₁₁H₅F₄NO₂([M + H]⁺): 258.0178. Found:

258.0176.

(E)-2-Cyano-3-phenylacrylic Acid (JXL048). ¹H NMR (500 MHz,

DMSO-d₆) δ 8.33 (s, 1H), 8.02 (m, 2H), 7.59 (m, 3H). ¹³C NMR

(126 MHz, DMSO-d₆) δ 163.7, 154.9, 133.6, 132.0, 131.1, 129.8,

116.5, 104.3. HRMS (ESI, m/z) calcd for C₁₀H₇NO₂([M − H]⁻):

172.0399. Found: 172.0396.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-5-ﬂuoro-1H-indol-3-yl)-

1

2-cyanoacrylic Acid (JXL038). H NMR (500 MHz, DMSO-d₆) δ

8.80 (s, 1H), 8.49 (s, 1H), 8.06 (s, 1H), 8.04 (s, 2H), 7.86 (dd, J =

9.6, 2.1 Hz, 1H), 7.69 (dd, J = 8.9, 4.3 Hz, 1H), 7.17 (dt, J = 9.0, 2.2

Hz, 1H), 5.83 (s, 2H). ¹³C NMR (126 MHz, DMSO-d₆) δ 164.8,

(E)-2-Cyano-3-(4-hydroxyphenyl)acrylic Acid (JXL049). ¹H NMR

(500 MHz, DMSO-d₆) δ 8.16 (s, 1H), 7.95 (d, J = 8.8 Hz, 2H), 6.92

(d, J = 8.8 Hz, 2H). ¹³C NMR (126 MHz, DMSO-d₆) δ 164.2, 163.0,

153.7, 133.6, 122.8, 117.2, 116.3, 99.2. HRMS (ESI, m/z) calcd for

C₁₀H₇NO₃([M − H]⁻): 188.0348. Found: 188.0346.

159.3 (d, J_c−f= 237.5 Hz), 145.9, 140.3, 136.2, 133.1, 131.1 (q, J_c−f

=

33.1 Hz), 129.1, 123.6 (q, J_c−f= 272.2 Hz), 118.4, 113.5, 112.7, 112.5,

110.4, 105.5, 105.3, 96.1, 49.6. HRMS (ESI, m/z) calcd for

C₂₁H₁₁F₇N₂O₂([M + H]⁺): 457.0787. Found: 457.0787.

(E)-2-Cyano-3-(1-(3,4-diﬂuorobenzyl)-6-ﬂuoro-1H-indol-3-yl)-

1

acrylic Acid (JXL039). H NMR (500 MHz, DMSO-d₆) δ 8.64 (s,

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-4-chloro-1H-indol-3-yl)-

1

1H), 8.46 (s, 1H), 7.99 (dd, J = 8.7, 5.1 Hz, 1H), 7.58 (dd, J = 9.8, 1.8

2-cyanoacrylic Acid (JXL050). H NMR (500 MHz, DMSO-d₆) δ

Hz, 1H), 7.43 (m, 2H), 7.12 (m, 2H), 5.57 (s, 2H). ¹³C NMR (126

9.20 (s, 1H), 8.90 (s, 1H), 8.06 (m, 3H), 7.72 (d, J = 7.5 Hz, 1H),

7.32 (m, 2H), 5.86 (s, 2H). ¹³C NMR (126 MHz, DMSO-d₆) δ 164.7,

146.7, 140.0, 138.0, 135.6, 131.1 (q, J_c−f= 33.1 Hz), 129.1, 125.6,

125.1, 124.7, 124.5, 123.7, 123.6 (q, J_c−f= 273.7 Hz), 122.5, 111.6,

109.9, 96.7, 49.6. HRMS (ESI, m/z) calcd for C₂₁H₁₁ClF₆N₂O₂([M

+ H]⁺): 473.0492. Found: 473.0494.

MHz, DMSO-d₆) δ 164.8, 160.3 (d, J_c−f= 239.3 Hz), 149.8 (dd, J_c−f

=

239.3, 25.2 Hz), 149.6 (dd, J_c−f= 246.3, 25.2 Hz), 145.8, 136.8, 135.3,

134.5, 125.1, 125.0, 124.6, 121.1, 118.5, 117.5, 111.3 (d, J_c−f= 23.9

Hz), 110.2, 98.7 (d, J_c−f= 26.5 Hz), 96.2, 49.4. HRMS (ESI, m/z)

calcd for C₁₉H₁₁F₃N₂O₂([M + H]⁺): 357.0851. Found: 457.0835.

(E)-2-Cyano-3-(1-(3,5-diﬂuorobenzyl)-6-ﬂuoro-1H-indol-3-yl)-

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-4-bromo-1H-indol-3-yl)-

1

acrylic Acid (JXL040). H NMR (500 MHz, DMSO-d₆) δ 8.65 (s,

2-cyanoacrylic Acid (JXL051). H NMR (500 MHz, DMSO-d₆) δ

1H), 8.46 (s, 1H), 8.00 (dd, J = 8.6, 5.2 Hz, 1H), 7.57 (d, J = 9.7 Hz,

1H), 7.15 (m, 2H), 7.03 (s, 1H), 7.02 (s, 1H), 5.62 (s, 2H). ¹³C

NMR (126 MHz, DMSO-d₆) δ 164.8, 163.0 (d, J_c−f= 239.4 Hz),

162.9 (d, J_c−f= 248.6 Hz), 160.4 (d, J_c−f= 239.4 Hz), 145.5, 141.3,

9.37 (s, 1H), 8.91 (s, 1H), 8.07 (app. s, 3H), 7.77 (d, J = 8.2 Hz, 1H),

7.51 (d, J = 7.6 Hz, 1H), 7.23 (t, J = 8.0 Hz, 1H), 5.85 (s, 2H). ¹³C

NMR (126 MHz, DMSO-d₆) δ 164.7, 146.3, 140.0, 138.0, 135.8,

131.0 (q, J_c−f= 33.1 Hz), 129.1, 127.9, 125.4, 124.9, 123.7 (q, J_c−f

=

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Article

273.2 Hz), 122.5, 118.2, 113.6, 122.1, 110.2, 96.6, 49.5. HRMS (ESI,

m/z) calcd for C₂₁H₁₁BrF₆N₂O₂([M + H]⁺): 518.9968. Found:

518.9948.

146.6, 145.7, 142.3, 136.6, 135.4, 130.7 (q, J_c−f= 32.8 Hz), 130.5,

128.8, 128.4, 128.0, 127.5, 125.7, 124.1, 123.6 (q, J_c−f= 273.7 Hz),

121.8, 118.3, 111.8, 110.3, 107.1, 96.2, 70.3, 52.4. HRMS (ESI, m/z)

calcd for C₂₈H₁₈F₆N₂O₃([M + H]⁺): 545.1300. Found: 545.1298.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-4-methoxy-1H-indol-3-

yl)-2-cyanoacrylic Acid (JXL060). ¹H NMR (500 MHz, DMSO-d₆) δ

8.99 (s, 1H), 8.71 (s, 1H), 8.05 (s, 1H), 8.00 (s, 2H), 7.24 (app. s,

2H), 6.82 (d, J = 6.0 Hz, 1H), 5.81 (s, 2H), 3.92 (s, 3H). ¹³C NMR

(126 MHz, DMSO-d₆) δ 165.0, 154.8, 148.6, 140.5, 137.8, 133.3,

131.0 (q, J_c−f= 32.8 Hz), 128.9, 125.4, 123.6 (q, J_c−f= 273.7 Hz),

122.5, 118.5, 116.8, 110.5, 105.0, 104.3, 95.3, 56.2, 49.5. HRMS (ESI,

m/z) calcd for C₂₂H₁₄F₆N₂O₃([M + H]⁺): 469.0987. Found:

469.0987.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-4-ﬂuoro-1H-indol-3-yl)-

1

2-cyanoacrylic Acid (JXL052). H NMR (500 MHz, DMSO-d₆) δ

8.80 (s, 1H), 8.58 (s, 1H), 8.05 (app. s, 3H), 7.54 (d, J = 8.2 Hz, 1H),

7.29 (d, J = 12.9 Hz, 1H), 7.09 (dd, J = 11.1, 8.2 Hz, 1H), 5.85 (s,

2H). ¹³C NMR (126 MHz, DMSO-d₆) δ 164.5, 155.7 (d, J_c−f= 239.4

Hz), 146.5, 140.1, 138.8, 134.5, 131.1 (q, J_c−f= 33.0 Hz), 129.0, 125.1

(d, J_c−f= 7.6 Hz), 124.7, 123.6 (q, J_c−f= 273.5 Hz), 122.5, 118.1,

116.1 (d, J_c−f= 18.5 Hz), 108.8, 108.5, 97.2, 49.7. HRMS (ESI, m/z)

calcd for C₂₁H₁₁F₇N₂O₂([M + H]⁺): 457.0787. Found: 457.0786.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-7-ﬂuoro-1H-indol-3-yl)-

1

2-cyanoacrylic Acid (JXL053). H NMR (500 MHz, DMSO-d₆) δ

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-5-bromo-1H-indol-3-yl)-

1

8.73 (br. s, 1H), 8.48 (br. s, 1H), 8.06 (br. s, 1H), 7.89 (br. s, 2H),

7.78 (br. s, J = 7.4 Hz, 1H), 7.22 (br. s, 1H), 7.10 (br. s, 1H), 5.89 (s,

2H). ¹³C NMR (126 MHz, DMSO-d₆) δ 164.6, 159.7 (d, J_c−f= 245.7

Hz), 145.6, 141.0, 136.2, 132.1, 130.0 (q, J_c−f= 33.0 Hz), 128.4,

123.9, 123.8, 123.6 (q, J_c−f= 273.3 Hz), 122.4 (d, J_c−f= 18.9 Hz),

118.1, 115.8, 110.6, 110.2 (d, J_c−f= 18.9 Hz), 97.3, 52.0. HRMS (ESI,

m/z) calcd for C₂₁H₁₁F₇N₂O₂([M + H]⁺): 457.0787. Found:

457.0792.

2-cyanoacrylic Acid (JXL061). H NMR (500 MHz, DMSO-d₆) δ

8.75 (s, 1H), 8.49 (s, 1H), 8.25 (s, 1H), 8.06 (s, 1H), 8.02 (m, 3H),

7.64 (app. s, 1H), 7.44 (app. s, 1H), 5.82 (s, 2H). ¹³C NMR (126

MHz, DMSO-d₆) δ 164.7, 145.3, 140.3, 135.5, 135.3, 131.1 (q, J_c−f

=

32.8 Hz), 129.8, 128.9, 127.0, 124.7, 123.6 (q, J_c−f= 274.0 Hz), 122.5,

122.3, 118.5, 113.9, 109.9, 97.7, 49.5. HRMS (ESI, m/z) calcd for

C₂₁H₁₁BrF₆N₂O₂([M + H]⁺): 516.9987. Found: 516.9983.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-6-bromo-1H-indol-3-yl)-

1

2-cyanoacrylic Acid (JXL062). H NMR (500 MHz, DMSO-d₆) δ

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-5-chloro-1H-indol-3-yl)-

1

2-cyanoacrylic Acid (JXL054). H NMR (500 MHz, DMSO-d₆) δ

8.72 (s, 1H), 8.47 (s, 1H), 8.06 (s, 2H), 8.03 (s, 1H), 7.94 (d, J = 8.2

Hz, 1H), 7.39 (d, J = 7.9 Hz, 1H), 5.81 (s, 2H). ¹³C NMR (126 MHz,

DMSO-d₆) δ 164.7, 145.6, 140.2, 137.4, 135.3, 131.1 (q, J_c−f= 32.8

Hz), 129.1, 127.0, 125.9, 123.6 (q, J_c−f= 274.0 Hz), 122.5, 121.5,

118.2, 117.2, 114.8, 110.3, 96.9, 49.3. HRMS (ESI, m/z) calcd for

C₂₁H₁₁BrF₆N₂O₂([M + H]⁺): 516.9987. Found: 516.9965.

8.78 (s, 1H), 8.51 (s, 1H), 8.12 (s, 1H), 8.06 (s, 1H), 8.03 (s, 2H),

7.70 (d, J = 7.4 Hz, 1H), 7.33 (d, J = 7.0 Hz, 1H), 5.83 (s, 2H). ¹³C

NMR (126 MHz, DMSO-d₆) δ 164.7, 145.7, 140.2, 135.9, 135.1,

131.1 (q, J_c−f= 33.0 Hz), 129.3, 129.0, 127.9, 124.4, 123.6 (q, J_c−f

=

274.0 Hz), 122.5, 119.5, 118.4, 113.6, 109.9, 96.9, 49.5. HRMS (ESI,

m/z) calcd for C₂₁H₁₁ClF₆N₂O₂([M + H]⁺): 473.0492. Found:

473.0498.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-6-chloro-1H-indol-3-yl)-

1

2-cyanoacrylic Acid (JXL063). H NMR (500 MHz, DMSO-d₆) δ

8.75 (s, 1H), 8.47 (s, 1H), 8.05 (m, 3H), 8.00 (d, J = 7.4 Hz, 1H),

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-4-cyano-1H-indol-3-yl)-

1

2-cyanoacrylic Acid (JXL055). H NMR (500 MHz, DMSO-d₆) δ

7.89 (s, 1H), 7.28 (d, J = 6.5 Hz, 1H), 5.81 (s, 2H). ¹³C NMR (126

9.03 (s, 1H), 9.00 (s, 1H), 8.09 (m, 4H), 7.80 (d, J = 7.3 Hz, 1H),

7.47 (t, J = 7.8 Hz, 1H), 5.90 (s, 2H). ¹³C NMR (126 MHz, DMSO-

d₆) δ 164.4, 144.4, 139.9, 136.7, 131.1 (q, J_c−f= 33.0 Hz), 129.7,

129.2, 126.6, 124.3, 123.6 (q, J_c−f= 273.7 Hz), 122.7, 118.6, 117.9,

117.8, 109.3, 101.7, 98.1, 49.5. HRMS (ESI, m/z) calcd for

C₂₂H₁₁F₆N₃O₂([M + H]⁺): 464.0834. Found: 464.0832.

(E)-1-(3,5-Bis(triﬂuoromethyl)benzyl)-3-(2-carboxy-2-cyanovin-

yl)-1H-indole-4-carboxylic Acid (JXL056). ¹H NMR (500 MHz,

DMSO-d₆) δ 9.26 (br. s, 1H), 8.88 (br. s, 1H), 8.05 (app. s, 3H), 7.94

(br. s, 1H), 7.76 (br. s, 1H), 7.37 (br. s, 1H), 5.85 (s, 2H). ¹³C NMR

(126 MHz, DMSO-d₆) δ 169.3, 165.0, 150.6, 140.2, 137.7, 136.0,

MHz, DMSO-d₆) δ 164.7, 145.6, 140.2, 137.0, 135.4, 131.1 (q, J_c−f=

32.8 Hz),129.2, 129.0, 126.7, 123.6 (q, J_c−f= 274.0 Hz), 123.3, 122.5,

121.2, 118.2, 111.9, 110.3, 97.0, 49.3. HRMS (ESI, m/z) calcd for

C₂₁H₁₁ClF₆N₂O₂([M + H]⁺): 473.0492. Found: 473.0493.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-7-chloro-1H-indol-3-yl)-

1

2-cyanoacrylic Acid (JXL064). H NMR (500 MHz, DMSO-d₆) δ

8.69 (s, 1H), 8.50 (s, 1H), 8.03 (s, 1H), 7.97 (d, J = 7.5 Hz, 1H), 7.73

(s, 2H), 7.32 (d, J = 7.1 Hz, 1H), 7.25 (m, 1H), 6.14 (s, 2H). ¹³C

NMR (126 MHz, DMSO-d₆) δ 164.5, 145.1, 142.1, 136.9, 131.5,

131.2 (q, J_c−f= 32.8 Hz), 127.6, 126.1, 124.1, 123.6 (q, J_c−f= 274.0

Hz), 122.5, 122.0, 118.8, 118.0, 116.9, 110.3, 97.9, 51.8. HRMS (ESI,

m/z) calcd for C₂₁H₁₁ClF₆N₂O₂([M + H]⁺): 473.0492. Found:

473.0496.

131.1 (q, J_c−f= 33.0 Hz), 129.0, 125.9, 125.2, 124.7, 123.6 (q, J_c−f

=

273.7 Hz), 123.5, 122.5, 118.3, 116.1, 110.1, 96.4, 49.3. HRMS (ESI,

m/z) calcd for C₂₂H₁₂F₆N₂O₄([M + H]⁺): 483.0779. Found:

483.0778.

Ethyl (E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-4-bromo-1H-

indol-3-yl)-2-cyanoacrylate (JXL065). ¹H NMR (500 MHz,

DMSO-d₆) δ 9.67 (s, 1H), 8.71 (s, 1H), 7.86 (s, 1H), 7.56 (s,

2H), 7.52 (d, J = 7.4 Hz, 1H), 7.17 (m, 2H), 5.55 (s, 2H), 4.37 (q, J =

7.1 Hz, 2H), 1.40 (t, J = 7.1 Hz, 3H). ¹³C NMR (126 MHz, DMSO-

d₆) δ 163.4, 146.9, 137.6, 137.4, 134.3, 132.7 (q, J_c−f= 33.9 Hz),

128.2, 126.7, 125.6, 125.1, 122.8 (q, J_c−f= 273.4 Hz), 122.7, 118.0,

114.9, 111.8, 109.9, 96.3, 62.2, 50.6, 14.3. HRMS (ESI, m/z) calcd for

C₂₃H₁₅BrF₆N₂O₂([M + H]⁺): 545.0299. Found: 545.0300.

Ethyl (E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-4-ﬂuoro-1H-indol-

(E)-3-(4-(Benzyloxy)-1-(3,5-bis(triﬂuoromethyl)benzyl)-1H-indol-

3-yl)-2-cyanoacrylic Acid (JXL057). ¹H NMR (500 MHz, DMSO-d₆)

δ 9.15 (s, 1H), 8.72 (s, 1H), 8.04 (s, 1H), 7.99 (s, 2H), 7.54 (br. s,

3H), 7.37 (br. s, 2H), 7.28 (m, 2H), 6.95 (s, 1H), 5.81 (s, 2H), 5.28

(s, 2H). ¹³C NMR (126 MHz, DMSO-d₆) δ 165.0, 153.7, 148.6,

140.5, 137.9, 133.4, 131.2 (q, J_c−f= 32.8 Hz), 128.9, 128.8, 128.1,

127.5, 125.4, 124.7, 123.6 (q, J_c−f= 273.7 Hz), 122.3, 118.5, 117.0,

110.5, 105.6, 105.2, 95.5, 70.0, 49.5. HRMS (ESI, m/z) calcd for

C₂₈H₁₈F₆N₂O₃([M + H]⁺): 545.1300. Found: 545.1299.

1

3-yl)-2-cyanoacrylate (JXL066). H NMR (500 MHz, DMSO-d₆) δ

8.86 (s, 1H), 8.61 (s, 1H), 7.86 (s, 1H), 7.56 (s, 2H), 7.23 (m, 1H),

7.01 (m, 2H), 5.54 (s, 2H), 4.37 (q, J = 7.1 Hz, 2H), 1.39 (t, J = 7.1

Hz, 3H). ¹³C NMR (126 MHz, DMSO-d₆) δ 163.1, 157.4 (d, J =

(E)-3-(6-(Benzyloxy)-1-(3,5-bis(triﬂuoromethyl)benzyl)-1H-indol-

3-yl)-2-cyanoacrylic Acid (JXL058). ¹H NMR (500 MHz, DMSO-d₆)

δ 8.63 (s, 1H), 8.43 (s, 1H), 8.05 (s, 1H), 8.03 (s, 2H), 7.86 (d, J =

8.5 Hz, 1H), 7.32 (m, 6H), 6.97 (d, J = 8.3 Hz, 1H), 5.77 (s, 2H),

5.09 (s, 2H). ¹³C NMR (126 MHz, DMSO-d₆) δ 165.0, 156.7, 146.1,

140.5, 137.5, 137.3, 134.3, 131.0 (q, J_c−f= 32.8 Hz), 128.9, 128.8,

128.3, 128.2, 126.9, 123.6 (q, J_c−f= 273.4 Hz), 122.4, 121.9, 118.5,

113.3, 110.5, 96.8, 95.6, 70.2, 49.2. HRMS (ESI, m/z) calcd for

C₂₈H₁₈F₆N₂O₃([M + H]⁺): 545.1300. Found: 545.1292.

(E)-3-(7-(Benzyloxy)-1-(3,5-bis(triﬂuoromethyl)benzyl)-1H-indol-

3-yl)-2-cyanoacrylic Acid (JXL059). ¹H NMR (500 MHz, DMSO-d₆)

δ 8.61 (br. s, 1H), 8.45 (br. s, 1H), 7.97 (br. s, 1H), 7.60 (br. s, 2H),

7.51 (br. s, 1H), 7.25 (br. s, 2H), 7.16 (br. s, 2H), 6.91 (br. s, 1H),

5.94 (s, 2H), 5.13 (s, 2H). ¹³C NMR (126 MHz, DMSO-d₆) δ 164.8,

243.9 Hz), 147.4, 138.2 (d, J = 10.1 Hz), 137.8, 133.2, 132.7 (q, J_c−f

=

33.9 Hz), 126.7, 125.3, 122.8 (q, J_c−f= 273.4 Hz), 122.7, 117.9, 116.9

(d, J_c−f= 17.6 Hz), 110.0, 109.0 (d, J_c−f= 19.5 Hz), 106.6, 97.1, 62.2,

50.7, 14.3. HRMS (ESI, m/z) calcd for C₂₃H₁₅F₇N₂O₂([M + H]⁺):

485.1100. Found: 485.1100.

(Z)-5-((1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-indol-3-yl)-

methylene)-2-iminothiazolidin-4-one (JXL067). ¹H NMR (500

MHz, DMSO-d₆) δ 9.21 (s, 1H), 9.01 (s, 1H), 8.03 (s, 1H), 7.94

(br. s, 3H), 7.86 (d, J = 7.7 Hz, 1H), 7.81 (s, 1H), 7.58 (d, J = 7.9 Hz,

1H), 7.24 (m, 1H), 7.19 (m, 1H), 5.76 (s, 2H). ¹³C NMR (126 MHz,

DMSO-d₆) δ 180.0, 174.9, 141.4, 136.5, 131.0 (q, J_c−f= 32.8 Hz),

2059

https://dx.doi.org/10.1021/acs.jmedchem.0c01570

J. Med. Chem. 2021, 64, 2046−2063

Journal of Medicinal Chemistry

pubs.acs.org/jmc

Article

130.4, 128.5, 127.9, 125.1, 123.8, 123.6 (q, J_c−f= 274.0 Hz), 122.1,

121.7, 120.8, 119.3, 111.8, 111.3, 48.9. HRMS (ESI, m/z) calcd for

C₂₁H₁₃F₆N₃OS ([M + H]⁺): 470.0762. Found: 470.0761.

DMSO-d₆) δ 164.4, 148.7, 145.8, 145.7, 140.2, 136.1, 135.3, 131.0 (q,

_c−f= 32.8 Hz), 129.5, 123.6 (q, J_c−f= 274.0 Hz), 122.4, 120.4, 117.7,

J

116.8, 108.2, 98.1, 48.2. HRMS (ESI, m/z) calcd for C₂₀H₁₀ClF₆N₃O₂

tert-Butyl (E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-indol-3-

([M + H]⁺): 474.0444. Found: 474.0448.

1

yl)-2-cyanoacrylate (JXL068). H NMR (500 MHz, CDCl₃) δ 8.57

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-4-bromo-1H-pyrrolo-

[2,3-b]pyridin-3-yl)-2-cyanoacrylic Acid (JXL077). ¹H NMR (500

MHz, DMSO-d₆) δ 9.17 (s, 1H), 9.02 (s, 1H), 8.27 (d, J = 5.0 Hz,

1H), 8.12 (s, 2H), 8.05 (s, 1H), 7.63 (d, J = 5.0 Hz, 1H), 5.85 (s,

2H). ¹³C NMR (126 MHz, DMSO-d₆-d₆) δ 164.4, 148.2, 145.5,

145.3, 140.2, 135.4, 131.0 (q, J_c−f= 32.8 Hz), 129.6, 124.9, 123.7,

123.6 (q, J_c−f= 274.0 Hz), 122.4, 118.4, 117.7, 108.6, 97.9, 48.2.

HRMS (ESI, m/z) calcd for C₂₀H₁₀BrF₆N₃O₂([M + H]⁺): 517.9939.

Found: 517.9935.

(s, 1H), 8.54 (s, 1H), 7.86 (d, J = 7.6 Hz, 1H), 7.84 (s, 1H), 7.56 (s,

2H), 7.33 (m, 2H), 7.22 (d, J = 7.4 Hz, 1H), 5.55 (s, 2H), 1.59 (s,

9H). ¹³C NMR (126 MHz, CDCl₃) δ 162.4, 144.7, 138.2, 135.9,

133.0, 132.7 (q, J_c−f= 32.8 Hz), 128.6, 126.7, 124.5, 123.0, 122.9 (q,

J_c−f= 277.2 Hz), 122.6, 119.0, 118.2, 111.1, 110.4, 97.7, 82.9, 50.4,

28.1. HRMS (ESI, m/z) calcd for C₂₅H₂₀F₆N₂O₂([M + H]⁺):

495.1507. Found: 495.1503.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-pyrrolo[2,3-b]-

pyridin-3-yl)-2-cyanoacrylic Acid (JXL069). ¹H NMR (500 MHz,

DMSO-d₆) δ 8.85 (s, 1H), 8.47 (m, 3H), 8.09 (s, 2H), 8.04 (s, 1H),

7.35 (dd, J = 7.1, 4.6 Hz, 1H), 5.84 (s, 2H). ¹³C NMR (126 MHz,

Methyl (E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-indol-3-yl)-2-

1

cyanoacrylate (JXL078). H NMR (500 MHz, CDCl₃) δ 8.62 (s,

1H), 8.61 (s, 1H), 7.89 (d, J = 7.7 Hz, 1H), 7.84 (s, 1H), 7.57 (s,

2H), 7.35 (m, 2H), 7.24 (m, 1H), 5.55 (s, 2H), 3.92 (s, 3H). ¹³C

NMR (126 MHz, CDCl₃) δ 164.5, 145.8, 138.1, 136.0, 133.5, 132.7

(q, J_c−f= 32.8 Hz), 128.6, 126.8, 124.7, 123.9, 123.6 (q, J_c−f= 274.0

Hz), 123.2, 119.1, 118.0, 111.2, 110.5, 95.4, 53.0, 50.5. HRMS (ESI,

m/z) calcd for C₂₂H₁₄F₆N₂O₂([M + H]⁺): 453.1038. Found:

453.1045.

DMSO-d₆) δ 164.6, 147.8, 146.1, 145.5, 141.0, 135.2, 131.0 (q, J_c−f

=

32.8 Hz), 129.4, 129.2, 123.6 (q, J_c−f= 274.0 Hz), 122.3, 120.1, 119.1,

118.2, 108.7, 97.1, 47.8. HRMS (ESI, m/z) calcd for C₂₀H₁₁F₆N₃O₂

([M + H]⁺): 440.0834. Found: 440.0834.

(Z)-5-((4-Fluoro-1-phenyl-1H-indol-3-yl)methylene)thiazolidine-

1

2,4-dione (JXL070). H NMR (500 MHz, DMSO-d₆) δ 12.45 (br. s,

1H), 8.11 (s, 1H), 7.83 (s, 1H), 7.65 (m, 4H), 7.52 (s, 1H), 7.32 (m,

2H), 7.12 (m, 1H). ¹³C NMR (126 MHz, DMSO-d₆) δ 168.0, 167.7,

156.9 (d, J_c−f= 245.7 Hz), 138.7 (d, J_c−f= 10.1 Hz), 137.9, 130.8,

130.5, 128.8, 125.5, 125.4, 124.3, 120.0, 116.2 (d, J_c−f= 18.9 Hz),

110.7, 108.5, 108.1 (d, J_c−f= 18.9 Hz). HRMS (ESI, m/z) calcd for

C₁₈H₁₁FN₂O₂S ([M + H]⁺): 339.0640. Found: 339.0610.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)phenethyl)-1H-indol-3-yl)-2-cya-

1

noacrylic Acid (JXL079). H NMR (500 MHz, CD₃OD) δ 8.51 (s,

1H), 8.14 (s, 1H), 7.83 (d, J = 7.3 Hz, 1H), 7.74 (s, 1H), 7.52 (m,

3H), 7.31 (m, 2H), 4.64 (t, J = 6.4 Hz, 2H), 3.35 (t, J = 6.3 Hz, 2H).

¹³C NMR (126 MHz, CD₃OD) δ 165.3, 145.3, 141.1, 136.1, 133.6,

131.3 (q, J_c−f= 32.8 Hz), 129.3, 128.3, 123.7, 123.3 (q, J_c−f= 272.5

Hz), 122.3, 120.3, 118.1, 117.6, 110.6, 109.6, 94.1, 47.8, 35.0. HRMS

(ESI, m/z) calcd for C₂₂H₁₄F₆N₂O₂([M + H]⁺): 453.1038. Found:

453.1031.

(Z)-5-((6-Fluoro-1-phenyl-1H-indol-3-yl)methylene)thiazolidine-

1

2,4-dione (JXL071). H NMR (500 MHz, DMSO-d₆) δ 12.42 (s,

1H), 8.06 (app. s, 2H), 7.85 (s, 1H), 7.67 (m, 2H), 7.62 (m, 2H),

7.50 (m, 1H), 7.31 (d, J = 9.6 Hz, 1H), 7.16 (t, J = 8.5 Hz, 1H). ¹³C

NMR (126 MHz, DMSO-d₆) δ 168.0, 167.7, 160.6 (d, J_c−f= 245.7

Hz), 138.0, 136.3 (d, J_c−f= 12.6 Hz), 131.0, 130.6, 128.5, 125.0,

Ethyl (E)-3-(1-(2-(3,5-Bis(triﬂuoromethyl)phenyl)acetyl)-1H-

indol-3-yl)-2-cyanoacrylate (JXL080). ¹H NMR (500 MHz,

CDCl₃) δ 8.90 (s, 1H), 8.52 (s, 1H), 8.49 (d, J = 8.1 Hz, 1H),

7.88 (s, 1H), 7.86 (s, 2H), 7.78 (d, J = 7.5 Hz, 1H), 7.47 (m, 2H),

4.50 (s, 2H), 4.41 (q, J = 7.1 Hz, 2H), 1.43 (t, J = 7.1 Hz, 3H). ¹³C

NMR (126 MHz, CDCl₃) δ 168.0, 162.2, 144.2, 135.6, 134.4, 132.3

(q, J_c−f= 33.6 Hz), 130.1, 129.8, 128.8, 127.8, 127.3, 125.5, 123.1 (q,

J_c−f= 273.3 Hz), 118.3, 117.1, 117.0, 115.9, 101.8, 62.8, 41.9, 14.3.

HRMS (ESI, m/z) calcd for C₂₄H₁₆F₆N₂O₃([M + H]⁺): 495.1143.

Found: 495.1136.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-indol-3-yl)-2-(mor-

pholine-4-carbonyl)acrylonitrile (JXL081). ¹H NMR (500 MHz,

CDCl₃) δ 8.48 (s, 1H), 8.34 (s, 1H), 7.84 (s, 2H), 7.56 (app. s, 2H),

7.32 (m, 2H), 7.22 (m, 1H), 5.54 (s, 2H), 3.77 (br. s, 8H). ¹³C NMR

(126 MHz, CDCl₃) δ 164.0, 145.2, 138.4, 135.8, 132.7 (q, J_c−f= 34.0

Hz), 131.8, 128.4, 126.8, 125.0, 124.5, 123.1 (q, J_c−f= 273.3 Hz),

122.8, 121.8, 119.0, 118.6, 111.4, 110.3, 98.1, 66.7, 50.3, 50.0. HRMS

(ESI, m/z) calcd for C₂₅H₁₉F₆N₃O₂([M + H]⁺): 508.1460. Found:

508.1459.

Ethyl (E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-pyrrolo[2,3-b]-

pyridin-3-yl)-2-cyanoacrylate (JXL082). ¹H NMR (500 MHz,

CDCl₃) δ 8.64 (s, 1H), 8.51 (s, 1H), 8.48 (dd, J = 4.6, 1.2 Hz,

1H), 8.21 (dd, J = 7.9, 1.2 Hz, 1H), 7.83 (s, 1H), 7.77 (s, 2H), 7.34

(dd, J = 7.9, 4.6 Hz, 1H), 5.69 (s, 2H), 4.38 (q, J = 7.1 Hz, 2H), 1.40

(t, J = 7.1 Hz, 3H). ¹³C NMR (126 MHz, CDCl₃) δ 163.3, 147.6,

145.7, 145.0, 138.5, 132.7, 132.3 (q, J_c−f= 33.6 Hz), 127.9, 127.7,

123.1 (q, J_c−f= 273.3 Hz), 122.5, 120.3, 119.0, 117.6, 109.4, 97.0,

62.3, 48.3, 14.3. HRMS (ESI, m/z) calcd for C₂₂H₁₅F₆N₃O₂([M +

H]⁺): 468.1147. Found: 468.1146.

124.8, 123.5, 121.3 (d, J_c−f= 10.0 Hz), 120.0, 112.6, 111.1 (d, J_c−f

=

18.9 Hz), 98.2 (d, J_c−f= 18.9 Hz). HRMS (ESI, m/z) calcd for

C₁₈H₁₁FN₂O₂S ([M + H]⁺): 339.0604. Found: 339.0601.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-pyrrolo[3,2-b]-

pyridin-3-yl)-2-cyanoacrylic Acid (JXL072). ¹H NMR (500 MHz,

DMSO-d₆) δ 8.94 (s, 1H), 8.68 (s, 1H), 8.54 (s, 1H), 8.11 (m, 4H),

7.35 (s, 1H), 5.87 (s, 2H). ¹³C NMR (126 MHz, DMSO-d₆) δ 164.5,

145.9, 144.2, 140.1, 135.9, 131.1 (q, J_c−f= 32.8 Hz), 129.6, 129.2,

124.7, 123.6 (q, J_c−f= 274.0 Hz), 122.5, 120.0, 119.5, 117.9, 110.1,

97.2, 49.8. HRMS (ESI, m/z) calcd for C₂₀H₁₁F₆N₃O₂([M + H]⁺):

440.0834. Found: 440.0834.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-pyrrolo[2,3-c]-

pyridin-3-yl)-2-cyanoacrylic Acid (JXL073). ¹H NMR (500 MHz,

DMSO-d₆) δ 9.14 (s, 1H), 8.96 (s, 1H), 8.55 (s, 1H), 8.40 (d, J = 5.1

Hz, 1H), 8.19 (s, 2H), 8.11 (app. s, 2H), 5.95 (s, 2H). ¹³C NMR (126

MHz, DMSO-d₆) δ 164.5, 145.5, 140.7, 139.9, 138.1, 134.5, 133.6,

131.1 (q, J_c−f= 32.8 Hz), 129.4, 127.2, 123.6 (q, J_c−f= 274.0 Hz),

122.7, 118.0, 114.6, 109.7, 97.9, 49.8. HRMS (ESI, m/z) calcd for

C₃₀H₁₁F₆N₃O₂([M + H]⁺): 440.0834. Found: 440.0837.

(Z)-5-Benzylidenethiazolidine-2,4-dione (JXL074). ¹H NMR (500

MHz, DMSO-d₆) δ 12.60 (br. s, 1H), 7.77 (s, 1H), 7.58 (app. d, J =

7.3 Hz, 2H), 7.51 (app. t, J = 7.4 Hz, 2H), 7.46 (m, 1H). ¹³C NMR

(126 MHz, DMSO-d₆) δ 168.4, 167.8, 133.5, 132.3, 130.9, 130.5,

129.8, 124.0. HRMS (ESI, m/z) calcd for C₁₀H₇NO₂S ([M − H]⁻):

204.0119. Found: 204.0122.

(Z)-5-((1H-Indol-3-yl)methylene)thiazolidine-2,4-dione (JXL075).

¹H NMR (500 MHz, DMSO-d₆) δ 12.28 (s, 1H), 12.11 (s, 1H), 8.03

Ethyl (Z)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-indol-3-yl)-2-

1

cyano-3-hydroxyacrylate (JXL083). H NMR (500 MHz, CDCl₃)

(s, 1H), 7.87 (d, J = 7.3 Hz, 1H), 7.72 (s, 1H), 7.48 (d, J = 7.6 Hz,

1H), 7.23 (m, 1H), 7.18 (m, 1H). ¹³C NMR (126 MHz, DMSO-d₆) δ

168.2, 167.8, 136.7, 129.1, 127.3, 125.0, 123.5, 121.5, 118.8, 116.7,

112.9, 110.9. HRMS (ESI, m/z) calcd for C₁₂H₈N₂O₂S ([M + H]⁺):

243.0228. Found: 243.0230.

δ 14.61 (s, 1H), 8.67 (s, 1H), 8.31 (dd, J = 7.0 1.3 Hz, 1H), 7.84 (s,

1H), 7.57 (s, 2H), 7.33 (m, 2H), 7.21 (d, J = 7.4 Hz, 1H), 5.53 (s,

2H), 4.40 (q, J = 7.1 Hz, 2H), 1.41 (t, J = 7.1 Hz, 3H). ¹³C NMR

(126 MHz, CDCl₃) δ 179.1, 172.2, 138.1, 136.0, 135.7, 132.7 (q, J_c−f

= 33.6 Hz), 126.9, 124.5, 123.7, 123.6, 122.6, 122.9 (q, J_c−f= 273.4

Hz), 121.8, 118.3, 110.1, 109.6, 73.1, 62.3, 50.4, 14.3. HRMS (ESI,

m/z) calcd for C₂₃H₁₆F₆N₂O₃([M + H]⁺): 483.1143. Found:

483.1145.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-4-chloro-1H-pyrrolo[2,3-

1

b]pyridin-3-yl)-2-cyanoacrylic Acid (JXL076). H NMR (500 MHz,

DMSO-d₆) δ 9.00 (s, 1H), 8.98 (s, 1H), 8.36 (s, 1H), 8.12 (s, 2H),

8.04 (br. s, 1H), 7.47 (br. s, 1H), 5.85 (s, 2H). ¹³C NMR (126 MHz,

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Ethyl (Z)-3-Acetoxy-3-(1-(3,5-bis(triﬂuoromethyl)benzyl)-1H-

indol-3-yl)-2-cyanoacrylate (JXL084). ¹H NMR (500 MHz,

CDCl₃) δ 8.56 (s, 1H), 7.90 (m, 1H), 7.85 (s, 1H), 7.61 (s, 2H),

7.32 (m, 2H), 7.24 (m, 1H), 5.51 (s, 2H), 4.29 (q, J = 7.1 Hz, 2H),

2.48 (s, 3H), 1.36 (t, J = 7.1 Hz, 3H). ¹³C NMR (126 MHz, CDCl₃)

δ 166.9, 161.4, 137.7, 136.2, 135.5, 132.8 (q, J_c−f= 33.9 Hz), 127.0,

126.7, 124.6, 123.6, 122.8 (q, J_c−f= 273.4 Hz), 122.7, 121.8, 117.7,

110.7, 109.8, 89.3, 61.8, 50.5, 29.7, 21.3, 14.2. HRMS (ESI, m/z)

calcd for C₂₅H₁₈F₆N₂O₄([M + H]⁺): 525.1249. Found: 525.1233.

Ethyl (Z)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-indol-3-yl)-3-

J = 7.1 Hz, 2H), 4.04 (s, 3H), 1.40 (t, J = 7.1 Hz, 3H). ¹³C NMR

(126 MHz, CDCl₃) δ 167.8, 163.5, 150.1, 137.8, 137.2, 134.9, 132.8

(q, J_c−f= 33.7 Hz), 126.7, 125.8, 125.1, 123.6, 122.8 (q, J_c−f= 273.4

Hz), 122.7, 121.7, 118.0, 114.6, 111.3, 96.4, 62.1, 52.6, 50.5, 14.3.

HRMS (ESI, m/z) calcd for C₂₅H₁₈F₆N₂O₄([M + H]⁺): 525.1249.

Found: 525.1238.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-4-(methoxycarbonyl)-

1

1H-indol-3-yl)-2-cyanoacrylic Acid (JXL092). H NMR (500 MHz,

CDCl₃) δ 9.09 (s, 1H), 8.58 (s, 1H), 7.75 (d, J = 7.5 Hz, 1H), 7.68 (s,

1H), 7.47 (s, 2H), 7.34 (d, J = 8.2 Hz, 1H), 7.20 (app. t, J = 7.9 Hz,

1H), 5.51 (s, 2H), 3.87 (s, 3H). ¹³C NMR (126 MHz, CDCl₃) δ

167.8, 165.1, 149.5, 138.2, 137.2, 134.8, 132.3 (q, J_c−f= 33.7 Hz),

126.8, 126.2, 125.5, 124.8, 123.3, 122.8 (q, J_c−f= 273.4 Hz), 122.3

118.2, 114.7, 110.9, 97.2, 52.4, 50.2. HRMS (ESI, m/z) calcd for

C₂₃H₁₄F₆N₂O₄([M + H]⁺): 497.0936. Found: 497.0939.

1

chloro-2-cyanoacrylate (JXL085). H NMR (500 MHz, CDCl₃) δ

7.91 (s, 1H), 7.85 (s, 1H), 7.77 (m, 1H), 7.65 (s, 2H), 7.63 (s, 1H),

7.30 (m, 2H), 5.48 (s, 2H), 4.18 (q, J = 7.1 Hz, 2H), 1.18 (t, J = 7.1

Hz, 3H). ¹³C NMR (126 MHz, CDCl₃) δ 161.0, 155.6, 138.1, 136.2,

135.4, 133.4, 132.7 (q, J_c−f= 33.7 Hz), 127.1, 126.6, 124.5, 124.3,

122.8 (q, J_c−f= 273.4 Hz), 121.5, 115.8, 122.1, 110.4, 102.5, 62.4,

50.0, 13.9. HRMS (ESI, m/z) calcd for C₂₃H₁₅ClF₆N₂O₂([M + H]⁺):

501.0804. Found: 501.0803.

Diethyl (E)-(2-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-indol-3-yl)-

1-cyanovinyl)phosphonate (JXL086). ¹H NMR (500 MHz,

CDCl₃) δ 8.55 (s, 1H), 8.33 (d, J = 19.7 Hz, 1H), 7.86 (d, J = 7.9

Hz, 1H), 7.83 (s, 1H), 7.58 (s, 2H), 7.31 (m, 2H), 7.22 (m, 1H), 5.54

(s, 2H), 4.21 (m, 4H), 1.39 (t, J = 7.0 Hz, 6H). ¹³C NMR (126 MHz,

CDCl₃) δ 149.8, 138.3, 135.8, 132.7 (q, J_c−f= 33.7 Hz), 132.6, 128.2,

126.8, 124.5, 123.0, 122.9 (q, J_c−f= 273.4 Hz), 122.5, 119.0, 117.8 (d,

Ethyl (E)-3-(4-Chloro-1-phenyl-1H-indol-3-yl)-2-cyanoacrylate

(JXL093). ¹H NMR (500 MHz, CDCl₃) δ 9.51 (s, 1H), 8.78 (s,

1H), 7.59 (m, 2H), 7.51 (m, 3H), 7.40 (dd, J = 8.3, 0.8 Hz, 1H), 7.32

(dd, J = 7.7, 0.8 Hz, 1H), 7.22 (app. t, J = 8.0 Hz, 1H), 4.38 (q, J = 7.1

Hz, 2H), 1.40 (t, J = 7.1 Hz, 3H). ¹³C NMR (126 MHz, CDCl₃) δ

163.6, 147.6, 138.1, 137.4, 134.2, 130.1, 129.0, 126.9, 125.3, 124.6,

124.5, 124.3, 118.0, 111.8, 110.6, 96.0, 62.1, 14.3. HRMS (ESI, m/z)

calcd for C₂₀H₁₅ClN₂O₂([M + H]⁺): 351.0900. Found: 351.0900.

(E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-indol-3-yl)-2-cya-

1

noacrylamide (JXL094). H NMR (500 MHz, CDCl₃) δ 8.70 (s,

J

_c−p= 11.3 Hz), 112.2 (d, J_c−p= 18.9 Hz), 110.3, 91.6 (d, J_c−p= 207.9

1H), 8.47 (s, 1H), 7.92 (d, J = 7.1 Hz, 1H), 7.85 (s, 1H), 7.58 (s,

2H), 7.34 (m, 2H), 7.23 (m, 1H), 5.55 (s, 2H). ¹³C NMR (126 MHz,

CDCl₃) δ 163.0, 144.8, 138.1, 136.0, 132.7 (q, J_c−f= 33.7 Hz), 132.5,

128.6, 126.8, 126.1, 124.6, 123.1, 122.9 (q, J_c−f= 273.4 Hz), 122.6,

119.3, 111.3, 110.3, 96.1, 50.4. HRMS (ESI, m/z) calcd for

C₂₁H₁₃F₆N₃O ([M + H]⁺): 438.1041. Found: 438.1046.

Hz), 63.2, 50.4, 16.3. HRMS (ESI, m/z) calcd for C₂₄H₂₁F₆N₂O₃P

([M + H]⁺): 531.1272. Found: 531.1274.

Ethyl (E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-4-chloro-1H-

pyrrolo[2,3-b]pyridin-3-yl)-2-cyanoacrylate (JXL087). ¹H NMR

(500 MHz, CDCl₃) δ 9.22 (s, 1H), 8.77 (s, 1H), 8.32 (d, J = 5.2

Hz, 1H), 7.83 (s, 1H), 7.77 (s, 2H), 7.31 (d, J = 5.2 Hz, 1H), 5.67 (s,

2H), 4.37 (q, J = 7.1 Hz, 2H), 1.39 (t, J = 7.1 Hz, 3H). ¹³C NMR

(126 MHz, CDCl₃) δ 163.0, 148.5, 146.2, 145.4, 138.1, 137.4, 133.1,

132.4 (q, J_c−f= 33.7 Hz), 128.0, 122.9 (q, J_c−f= 273.4 Hz), 122.6,

121.8, 117.6, 117.2, 109.7, 97.8, 62.4, 48.6, 14.3. HRMS (ESI, m/z)

calcd for C₂₂H₁₄ClF₆N₃O₂([M + H]⁺): 502.0757. Found: 502.0756.

Ethyl (E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-4-bromo-1H-

pyrrolo[2,3-b]pyridin-3-yl)-2-cyanoacrylate (JXL088). ¹H NMR

(500 MHz, CDCl₃) δ 9.40 (s, 1H), 8.78 (s, 1H), 8.22 (d, J = 5.1

Hz, 1H), 7.83 (s, 1H), 7.77 (s, 2H), 7.49 (d, J = 5.1 Hz, 1H), 5.67 (s,

2H), 4.37 (q, J = 7.1 Hz, 2H), 1.39 (t, J = 7.1 Hz, 3H). ¹³C NMR

(126 MHz, CDCl₃) δ 163.1, 148.1, 145.7, 145.1, 138.1, 137.4, 133.4,

132.4 (q, J_c−f= 33.7 Hz), 123.7, 122.9 (q, J_c−f= 273.4 Hz), 122.6,

119.7, 117.6, 110.0, 101.4, 97.4, 62.4, 48.6, 14.3. HRMS (ESI, m/z)

calcd for C₂₂H₁₄BrF₆N₃O₂([M + H]⁺): 546.0252. Found: 546.0238.

Ethyl (E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-5-chloro-1H-indol-

3-yl)-2-cyanoacrylate (JXL089). ¹H NMR (500 MHz, CDCl₃) δ 8.60

(s, 1H), 8.50 (s, 1H), 7.85 (s, 1H), 7.83 (s, 1H), 7.55 (s, 2H), 7.28

(dd, J = 8.7, 1.7 Hz, 1H), 7.14 (d, J = 8.7 Hz, 1H), 5.54 (s, 2H), 4.38

(q, J = 7.1 Hz, 2H), 1.40 (t, J = 7.1 Hz, 3H). ¹³C NMR (126 MHz,

CDCl₃) δ 163.2, 144.8, 137.7, 134.3, 134.1, 132.8 (q, J_c−f= 33.7 Hz),

129.7, 129.3, 126.7, 125.1, 122.8 (q, J_c−f= 273.4 Hz), 122.7, 118.9,

117.7, 111.6, 110.6, 96.9, 62.3, 50.6, 14.3. HRMS (ESI, m/z) calcd for

C₂₃H₁₅ClF₆N₂O₂([M + H]⁺): 501.0804. Found: 501.0801.

Ethyl (E)-3-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-4-cyano-1H-indol-

3-yl)-2-cyanoacrylate (JXL090). ¹H NMR (500 MHz, CDCl₃) δ 9.29

(s, 1H), 8.78 (s, 1H), 7.87 (s, 1H), 7.68 (dd, J = 7.4, 0.8 Hz, 1H),

7.56 (s, 2H), 7.49 (dd, J = 8.4, 0.8 Hz, 1H), 7.38 (dd, J = 8.4, 7.4 Hz,

1H), 5.62 (s, 2H), 4.38 (q, J = 7.1 Hz, 2H), 1.40 (t, J = 7.1 Hz, 3H).

¹³C NMR (126 MHz, CDCl₃) δ 162.6, 144.3, 137.3, 136.2, 135.1,

132.9 (q, J_c−f= 33.7 Hz), 129.4, 127.3, 126.7, 124.1, 123.0, 122.8 (q,

J_c−f= 273.4 Hz), 117.9, 117.5, 115.4, 110.7, 103.5, 98.6, 62.4, 50.6,

14.3. HRMS (ESI, m/z) calcd for C₂₄H₁₅F₆N₃O₂([M + H]⁺):

492.1147. Found: 492.1149.

Methyl (E)-1-(3,5-Bis(triﬂuoromethyl)benzyl)-3-(2-cyano-3-

ethoxy-3-oxoprop-1-en-1-yl)-1H-indole-4-carboxylate (JXL091).

¹H NMR (500 MHz, CDCl₃) δ 9.36 (s, 1H), 8.71 (s, 1H), 7.93

(dd, 1H, J = 7.4, 1.1 Hz, 1H), 7.85 (s, 1H), 7.55 (s, 2H), 7.40 (dd, J =

8.3, 1.1 Hz, 1H), 7.34 (dd, J = 8.3, 7.4 Hz, 1H), 5.58 (s, 2H), 4.37 (q,

Diethyl (E)-(2-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-pyrrolo[2,3-

b]pyridin-3-yl)-1-cyanovinyl)phosphonate (JXL095). ¹H NMR (500

MHz, CDCl₃) δ 8.58 (s, 1H), 8.46 (dd, J = 4.7, 1.4 Hz, 1H), 8.24 (d, J

= 19.5 Hz 1H), 8.20 (dd, J = 8.0, 1.4 Hz, 1H), 7.82 (s, 1H), 7.77 (s,

2H), 7.32 (dd, J = 8.0, 4.7 Hz, 1H), 5.67 (s, 2H), 4.22 (m, 4H), 1.40

(t, J = 7.1 Hz, 6H). ¹³C NMR (126 MHz, CDCl₃) δ 149.2 (d, J_c−p

=

8.2 Hz), 147.4, 145.6, 138.6, 132.4 (q, J_c−f= 33.7 Hz), 132.0, 127.9,

127.8, 123.0 (q, J_c−f= 273.4 Hz), 122.4, 119.9, 118.9, 117.4 (d, J_c−p

=

11.3 Hz), 110.5 (d, J_c−p= 19.5 Hz), 92.8 (d, J_c−p= 205.1 Hz), 63.4,

48.3, 16.3. HRMS (ESI, m/z) calcd for C₂₃H₂₀F₆N₃O₃P ([M + H]⁺):

532.1225. Found: 532.1238.

(E)-(2-(1-(3,5-Bis(triﬂuoromethyl)benzyl)-1H-indol-3-yl)-1-

cyanovinyl)phosphonic Acid (JXL096). ¹H NMR (500 MHz,

CD₃OD) δ 8.58 (s, 1H), 8.25 (d, J = 19.6 Hz, 1H), 7.90 (s, 1H),

7.87 (m, 1H), 7.76 (s, 2H), 7.45 (m, 1H), 7.31 (m, 2H), 5.75 (s, 2H).

¹³C NMR (126 MHz, CD₃OD) δ 146.7 (J_c−p= 7.2 Hz), 140.2, 136.1,

132.1, 131.9 (q, J_c−f= 33.7 Hz), 128.0, 127.2, 123.8, 122.3, 123.2 (q,

J

_c−f= 273.4 Hz), 121.4, 118.2, 117.4 (d, J_c−p= 11.3 Hz), 111.5 (J_c−p=

18.4 Hz), 110.5, 94.6 (d, J_c−p= 201.2 Hz), 49.1. HRMS (ESI, m/z)

calcd for C₂₀H₁₃F₆N₂O₃P ([M + H]⁺): 475.0646. Found: 475.0642.

In Vitro Evaluation of the Analogues’ Activity in Promoting

Lactate Production Rate. Human breast epithelial (MCF10A) cells

were obtained from the American Type Culture Collection and

cultured in Dulbecco’s modiﬁed Eagle medium (DMEM):F12 media

supplemented with 5% of horse serum, 50 U/mL of penicillin−

streptomycin, 10 μg/mL of insulin, 0.5 μg/mL of hydrocortisone, 20

ng/mL of EGF, and 10 μg/mL of cholera toxin. To determine

whether the analogues could promote cellular lactate production,

MCF10A cells were treated with analogues and lactate concentrations

were measured using a BioProﬁle Basic Analyzer (Nova Biomedical).

Brieﬂy, MCF10A cells were seeded in 6-well plates in triplicate,

treated the following day with 10 μM analogue, or an equivalent

volume of DMSO, for 24−30 h. Culture media lactate concentrations

were measured and normalized to cell number and duration of the

experiment to calculate a cellular lactate production rate (nmol lactate

per million cells per hour). Ordinary one-way ANOVA followed by

Dunnett’s post-test was performed using GraphPad Prism 8.3.0,

comparing each analogue to DMSO.

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A Seahorse XF Analyzer (Agilent) was used to measure the oxygen

consumption rate (OCR) to calculate IC₅₀values for several of the

analogues. MCF10A cells were seeded in 96-well plates, treated with

half-log doses of analogue, ranging from 3 nM to 10 μM, in the

presence of 5 mM of pyruvate and 0.5 mM of malate. OCR (fmol O₂

per cell per min) was measured in duplicate and normalized to cell

number. IC₅₀values were calculated using Prism (GraphPad).

In Vivo Evaluation of the Selected Analogues’ Activity in

Accelerating Hair Growth on Mice. To determine the eﬃcacy of the

compounds on the hair cycle, wild-type C57BL/6J mice were shaved

at postnatal day 50 and topically treated with the compounds

disclosed herein suspended in lotion (20 μM) every other day for up

to 3 weeks. The lotion used was a Transderma Plo Ultramax gel. The

mice were treated topically every other day with ∼300 μL at 20 μM.

Photographs were taken every week to monitor pigmentation and hair

growth.

https://pubs.acs.org/10.1021/acs.jmedchem.0c01570

Author Contributions

The manuscript was written and edited by Drs. X.L. and M.E.J.

All authors have given approval to the ﬁnal version of the

manuscript.

Notes

The authors declare no competing ﬁnancial interest.

ACKNOWLEDGMENTS

■

Dr. X.L. is a postdoctoral fellow supported by the UCLA

Tumor Cell Biology Training Program (USHHS Ruth L.

Kirschstein Institutional National Research Service Award no.

T32 CA009056). We thank Linsey Stiles at the UCLA

Mitochondria Core for her help with the Seahorse assays.

All mice were kept under deﬁned pathogen-free conditions at the

AAALAC-approved animal facility of the Division of Laboratory

Animals (DLAM) at UCLA. All animal experiments were performed

with the approval of the UCLA Oﬃce of Animal Resource Oversight

(OARO); the approval number is ARC no. 2016-055-11A.

ABBREVIATIONS

■

MPC, mitochondrial pyruvate carrier; LDH, lactate dehydro-

genase; TZD, thiazolidinedinone; OCR, oxygen consumption

rate; DMSO, dimethyl sulfoxide

ASSOCIATED CONTENT

* Supporting Information

The Supporting Information is available free of charge at

https://pubs.acs.org/doi/10.1021/acs.jmedchem.0c01570.

■

sı

REFERENCES

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Molecular formula strings (CSV)

AUTHOR INFORMATION

Corresponding Author

■

Michael E. Jung − Department of Chemistry and Biochemistry

and Department of Molecular and Medical Pharmacology,

UCLA, Los Angeles, California 90095, United States;

orcid.org/0000-0003-1290-8588; Phone: (310) 825-

7954; Email: jung@chem.ucla.edu

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Authors

Xiaoguang Liu − Department of Chemistry and Biochemistry,

UCLA, Los Angeles, California 90095, United States

Aimee A. Flores − Department of Molecular Cell and

Developmental Biology, Eli and Edythe Broad Center for

Regenerative Medicine, and Molecular Biology Institute,

UCLA, Los Angeles, California 90095, United States

Lisa Situ − Department of Molecular Cell and Developmental

Biology and Department of Biological Chemistry, UCLA, Los

Angeles, California 90095, United States

Wen Gu − Department of Molecular and Medical

Pharmacology, UCLA, Los Angeles, California 90095, United

States

Hui Ding − Department of Chemistry and Biochemistry,

UCLA, Los Angeles, California 90095, United States

Heather R. Christofk − Department of Molecular and

Medical Pharmacology, Jonsson Comprehensive Cancer

Center, Eli and Edythe Broad Center for Regenerative

Medicine, and Department of Biological Chemistry, UCLA,

Los Angeles, California 90095, United States

William E. Lowry − Department of Molecular Cell and

Developmental Biology, Jonsson Comprehensive Cancer

Center, Eli and Edythe Broad Center for Regenerative

Medicine, and Molecular Biology Institute, UCLA, Los

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J. Med. Chem. 2021, 64, 2046−2063

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M. I.; Ciaraldi, T. P.; Murphy, A. N. Thiazolidinediones Are Acute,

Specific Inhibitors of The Mitochondrial Pyruvate Carrier. Proc. Natl.

Acad. Sci. U. S. A. 2013, 110, 5422−5427.

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https://dx.doi.org/10.1021/acs.jmedchem.0c01570

J. Med. Chem. 2021, 64, 2046−2063

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Development of Novel Mitochondrial Pyruvate Carrier Inhibitors to Treat Hair Loss

DOI: 10.1021/acs.jmedchem.0c01570

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