The discovery of quinoline-3-carboxamides as hematopoietic prostaglandin D synthase (H-PGDS) inhibitors

Article abstract of DOI:10.1016/j.bmc.2019.02.017

With the goal of discovering more selective anti-inflammatory drugs, than COX inhibitors, to attenuate prostaglandin signaling, a fragment-based screen of hematopoietic prostaglandin D synthase was performed. The 76 crystallographic hits were sorted into similar groups, with the 3-cyano-quinoline 1a (FP IC₅₀ = 220,000 nM, LE = 0.43) being a potent member of the 6,6-fused heterocyclic cluster. Employing SAR insights gained from structural comparisons of other H-PGDS fragment binding mode clusters, the initial hit 1a was converted into the 70-fold more potent quinoline 1d (IC₅₀ = 3,100 nM, LE = 0.49). A systematic substitution of the amine moiety of 1d, utilizing structural information and array chemistry, with modifications to improve inhibitor stability, resulted in the identification of the 300-fold more active H-PGDS inhibitor tool compound 1bv (IC₅₀ = 9.9 nM, LE = 0.42). This selective inhibitor exhibited good murine pharmacokinetics, dose-dependently attenuated PGD₂ production in a mast cell degranulation assay and should be suitable to further explore H-PGDS biology.

Full text of DOI:10.1016/j.bmc.2019.02.017

Accepted Manuscript
The discovery of quinoline-3-carboxamides as hematopoietic prostaglandin D
synthase (H-PGDS) inhibitors
David N. Deaton, Young Do, Jason Holt, Michael R. Jeune, H. Fritz Kramer,
Andrew L. Larkin, Lisa A. Orband-Miller, Gregory E. Peckham, Chuck Poole,
Daniel J. Price, Lee T. Schaller, Ying Shen, Lisa M. Shewchuk, Eugene L.
Stewart, J. Darren Stuart, Stephen A. Thomson, Paris Ward, Joseph W. Wilson,
Tianshun Xu, Jeffrey H. Guss, Caterina Musetti, Alan R. Rendina, Karen
Affleck, David Anders, Ashley P. Hancock, Heather Hobbs, Simon T. Hodgson,
Jonathan Hutchinson, Melanie V. Leveridge, Harry Nicholls, Ian E. D. Smith,
Don O. Somers, Helen F. Sneddon, Sorif Uddin, Anne Cleasby, Paul N.
Mortenson, Caroline Richardson, Gordon Saxty
PII:
S0968-0896(18)32008-X
https://doi.org/10.1016/j.bmc.2019.02.017
BMC 14753
DOI:
Reference:
Bioorganic & Medicinal Chemistry
To appear in:
Received Date:
Revised Date:
Accepted Date:
14 December 2018
30 January 2019
8 February 2019
Please cite this article as: Deaton, D.N., Do, Y., Holt, J., Jeune, M.R., Fritz Kramer, H., Larkin, A.L., Orband-Miller,
L.A., Peckham, G.E., Poole, C., Price, D.J., Schaller, L.T., Shen, Y., Shewchuk, L.M., Stewart, E.L., Darren Stuart,
J., Thomson, S.A., Ward, P., Wilson, J.W., Xu, T., Guss, J.H., Musetti, C., Rendina, A.R., Affleck, K., Anders, D.,
Hancock, A.P., Hobbs, H., Hodgson, S.T., Hutchinson, J., Leveridge, M.V., Nicholls, H., E. D. Smith, I., Somers,
D.O., Sneddon, H.F., Uddin, S., Cleasby, A., Mortenson, P.N., Richardson, C., Saxty, G., The discovery of
quinoline-3-carboxamides as hematopoietic prostaglandin D synthase (H-PGDS) inhibitors, Bioorganic &
Medicinal Chemistry (2019), doi: https://doi.org/10.1016/j.bmc.2019.02.017
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The discovery of quinoline-3-carboxamides as hematopoietic prostaglandin D synthase (H-PGDS) inhibitors
David N. Deaton,a,* Young Do,a Jason Holt,a Michael R. Jeune,a H. Fritz Kramer,a Andrew L. Larkin,a Lisa A.
Orband-Miller,a Gregory E. Peckham,a Chuck Poole,a Daniel J. Price,a Lee T. Schaller,a Ying Shen,a Lisa M.
Shewchuk,a Eugene L. Stewart,a J. Darren Stuart,a Stephen A. Thomson,a Paris Ward,a Joseph W. Wilson,a Tianshun
Xu,a Jeffrey H. Guss,b Caterina Musetti,b Alan R. Rendina,b Karen Affleck,c David Anders,c Ashley P. Hancock,c
Heather Hobbs,c Simon T. Hodgson,c Jonathan Hutchinson,c Melanie V. Leveridge,c Harry Nicholls,c Ian E. D.
Smith,c Don O. Somers,c Helen F. Sneddon,c Sorif Uddin,c Anne Cleasby,d Paul N. Mortenson,d Caroline
Richardson,d Gordon Saxty,d
aGlaxoSmithKline, 5 Moore Drive, P.O. Box 13398, Research Triangle Park, NC 27709, USA
bGlaxoSmithKline, 1250 South Collegeville Road, Collegeville, PA 19426, USA
cGlaxoSmithKline, Gunnels Wood Road, Stevenage, Hertfordshire SG1 2NY, UK
dAstex Pharmaceuticals, 436 Cambridge Science Park, Milton Road, Cambridge CB4 0QA, UK
*Corresponding author. E-mail address: david.n.deaton@gsk.com.
KEYWORDS: Prostaglandin D2, PGD2, Hematopoietic prostaglandin D synthase, H-PGDS, H-PGDS inhibitor,
Fragment-based drug discovery
Abstract
With the goal of discovering more selective anti-inflammatory drugs, than COX inhibitors, to attenuate
prostaglandin signaling, a fragment-based screen of hematopoietic prostaglandin D synthase was performed. The 76
crystallographic hits were sorted into similar groups, with the 3-cyano-quinoline 1a (FP IC50 = 220,000 nM, LE =
0.43) being a potent member of the 6,6-fused heterocyclic cluster. Employing SAR insights gained from structural
comparisons of other H-PGDS fragment binding mode clusters, the initial hit 1a was converted into the 70-fold
more potent quinoline 1d (IC50 = 3,100 nM, LE = 0.49). A systematic substitution of the amine moiety of 1d,
utilizing structural information and array chemistry, with modifications to improve inhibitor stability, resulted in the
identification of the 300-fold more active H-PGDS inhibitor tool compound 1bv (IC50 = 9.9 nM, LE = 0.42). This
selective inhibitor exhibited good murine pharmacokinetics, dose-dependently attenuated PGD2 production in a mast
cell degranulation assay and should be suitable to further explore H-PGDS biology.
1.1 Introduction
The prostaglandins (PGs) are physiologically active, cyclic, oxygenated lipid compounds that are synthesized in the
cell from the polyunsaturated fatty acid arachidonic acid, which is liberated from the sn2 position of
glycerophospholipids by phospholipase A2.1 Cyclooxygenases 1 or 2 (COX 1 & 2, prostaglandin endoperoxide
synthases 1 & 2 (PGHS1 & 2)) insert two O2 molecules into arachidonic acid to produce prostaglandin G2 (PGG2),
and subsequently reduce PGG2 to PGH2. Various synthases then convert PGH2 into PGD2, PGE2, PGF2α, PGI2, and
thromboxane A2 (TxA2). These prostanoids are then secreted by the multidrug resistance protein 4 (MRP4,
ABCC4),2 where they act as autocrine or paracrine factors on adjacent cells via signaling through various seven
transmembrane G-protein-coupled receptors.3 They can subsequently be taken back up by cells via the
prostaglandin transporter (PGT, solute carrier organic ion transporter 2A1 (SLCO2A1), organic anion transporting
polypeptide 2A1 (OATO2A1))4 and inactivated by the enzyme 15-hydroxyprostaglandin dehydrogenase.5
PGD2 is produced by hematopoietic and/or lipocalin prostaglandin D synthases (EC 5.3.99.2, H-PGDS and L-
PGDS) from PGH2 by isomerization. It can be reduced by 11-keto PGD2 reductase (aldo-keto reductase 1C3
(AKR1C3)) to 9α, 11β-PGF2.6 PGD2 can also dehydrate to form PGJ2, Δ12-PGJ2, and/or 15-deoxy-Δ12,14-PGJ2.
PGD2 signals through the seven transmembrane receptors DP1 (Gs-coupled, stimulates cAMP production and Ca+2
release) and DP2 (Chemoattractant Receptor-homologous molecule expressed on Th2 cells receptor (CRTH2), Gi-
coupled, mobilizes Ca+2). PGD2 is a major prostaglandin produced by mast cells and helps mediate the
inflammatory responses of type 2 helper T lymphocytes (Th2 cells), mast cells, eosinophils, and dendritic cells.
PGD2 induces chemotaxis and stimulates cytokine production and has been implicated in allergic asthma,7 atopic

dermatitis,8,9 niacin-induced vasodilation,10 inflammatory bowel disease,11 and hair growth inhibition.12,13 The
physiological activity of PGD2 could be selectively attenuated by either antagonizing its interaction with any or all
its receptors, inhibiting either or both of its synthases, or enhancing its degradation.
H-PGDS is a member of the σ-class of glutathione-S-transferases.14 Its fellow enzyme, L-PGDS, is a member of the
lipocalin gene family;15 these structurally different proteins are an example of functional convergence.
Consequently, inhibitors of one enzyme may not be expected to bind to and interfere with the other enzyme’s
activity (vide infra). H-PGDS is expressed in dendritic cells and Langerhans cells of skin, Kupffer cells in the liver,
dendritic cells in the thymus, microglia in the brain, the oviduct, and megakaryocytes in the bone marrow, as well as
in mast cells, eosinophils, and activated Th2 cells in many tissues. H-PGDS has been implicated in asthma,16
allergic rhinitis,17, Krabbe disease,18 Duchenne muscular dystrophy,19 and lupus.20
Consequently, there has been strong interest in the discovery of H-PGDS inhibitors as potential treatments of these
conditions, with several research groups, including Osaka (HQL-79),21 Evotec,22 AstraZeneca,23 Pfizer,24 and Sanofi
(SAR191801B)25 disclosing their efforts in the primary literature. There is also an excellent review of H-PGDS
inhibitors from 2012.26 The structures of several representative H-PGDS tool compounds are depicted in Figure 1.
Recently, Taiho began a phase I clinical trial of a H-PGDS inhibitor TAS-205 in Duchenne muscular dystrophy
patients.27 Although, the structure of TAS-205 has not been identified, the structure may be the one shown below
from a recent Taiho patent application.28 With the many difficulties in translating efficacious tool compounds into
safe and effective drugs in humans, multiple H-PGDS chemotypes entering clinical trials will maximize the chance
for successful delivery of a medicine to aid patients with aberrant PGD2 production, thus requiring addition efforts
to identify suitable H-PGDS inhibitors for clinical investigation.
Figure 1. H-PGDS tool compounds.
N
N
N
O
HN
F F
HN
F
N
N
N
O
F
HQL-79
O
Pfizer
O
O
N
S
O
S
N
N
O
NH2
N
H
OH
O
N
N
N
HN
Evotec
Sanofi
N
O
N
N
N
O
S
N
N
O
HN
N
N
H
AstraZeneca
Taiho
As part of a larger drug discovery collaboration between GlaxoSmithKline and Astex Pharmaceuticals, a fragment-
based screening approach was initiated to discover inhibitors of H-PGDS for the treatment of various inflammatory
diseases.29,30 In comparison to high throughput screens, fragment-based screening campaigns sample chemical
space more efficiently, are cheaper to screen, are simpler to triage, and often provide more drug-like starting points.
This screening campaign resulted in the generation of 76 crystallographic hits, which were clustered and classified
based on their protein interactions and available growth vectors for chemistry. Hits from two of these clusters were
recently described.31 Another cluster consisting of 6,6-fused heterocycles contained the commercially available 3-
cyano-quinoline 1a. It was a weak, competitive, reversible inhibitor of H-PGDS in a fluorescence polarization assay
(FP IC50 = 220,000 nM), but exhibited an excellent ligand efficiency (LE = 0.43).32
Another X-ray hit from this 6,6-fused heterocycle cluster was 8-methoxychroman-3-carbonitrile. Although
synthetically less tractable, this fragment suggested that addition of an 8-methoxy moiety to quinoline 1a might be
beneficial to inhibitor potency. Furthermore, it could be inferred from another X-ray hit, 6-methoxy-1H-
benzo[d]imidazole, from a different cluster, consisting of 5,6-fused heterocycles, which bound similarly to the
quinoline 1a, that addition of a 7-methoxy group to the quinoline hit might also enhance H-PGDS inhibition. These
SAR insights gained from the binding mode clustering approach of these X-ray structures were applied to the
quinoline template. The commercially available quinolines were purchased and tested. The 7- and 8-

methoxyquinoline-3-carbonitriles 1b (FP IC50 = 13,000 nM) and 1c (FP IC50 = 5,000 nM) were over a log more
potent than nitrile 1a. This increase in inhibitory activity led to these compounds showing activity in a RapidFire™
mass spectrometry assay measuring the inhibition of the conversion of PGH2, generated in situ by COX-2 from
arachidonic acid, to PGD2 by H-PGDS (1b (IC50 = 13,000 nM, LE = 0.49) and 1c (IC50 = 8,400 nM, LE = 0.51).
Both compounds maintained good ligand efficiencies.
As shown in Figure 2, the co-crystal structure of fragment 1a bound to rat H-PGDS (rH-PGDS) and glutathione
(GSH) revealed that the quinoline ring forms a face to face π-π stacking interaction with the indole side chain of
104Trp (face of indole 104Trp to face of quinoline 1a distance = 3.6 Å). The coplanar 3-nitrile group facilitates this
interaction, withdrawing electron density from the quinoline ring and improving the interaction with the π-cloud of
the indole ring. Also, the quinoline nitrogen of 1a forms a hydrogen bond with a tightly bound, structural water
molecule (structural water oxygen to quinoline nitrogen distance = 2.9 Å) that is held in place via hydrogen bonds to
the C-terminal carboxylic acid of 199Leu, the hydroxyl side chain of 159Thr, and another bound water molecule. This
binding mode is similar to the Pfizer and Sanofi series, mimicking their pyridine and pyrimidine nitrogen
interactions with the protein. The 7- and 8-methoxy groups likely strengthen this interaction by increasing the
basicity of the quinoline nitrogen.
Furthermore, hit to lead efforts on the chroman series showed that substituted 3-carboxamides replacements of the
nitrile enhanced potency and these learnings were utilized in the quinoline series. As shown in Table 1, the 7-
methoxy-N-methylquinoline-3-carboxamide 1d (IC50 = 3,100 nM, LE = 0.49) was a 4-fold more potent inhibitor of
H-PGDS than its corresponding nitrile 1b, while the 8-methoxy-N-methylquinoline-3-carboxamide 1e (IC50 = 7,700
nM, LE = 0.45) was equipotent with its nitrile 1c. Structural analysis showed that a vector emanating from this 3-
position points toward a channel that exits to bulk water and would be an excellent position from which to append
groups on the quinoline core to enhance binding or improve drug properties. Thus, a hit-to-lead campaign strategy
was undertaken to explore the 3-position of the 7-methoxyquinoline ring via array chemistry by coupling various
amines to quinoline-3-carboxylic acids. The chemistry goal was to improve the potency of these fragments, while
maintaining ligand efficiency.
Figure 2. Co-crystal structure of inhibitor 1a bound to rH-PGDS with GSH.
Figure 2 Legend. Ligand binding site of the X-ray co-crystal structure of 1a complexed with rH-PGDS and GSH. The rH-PGDS carbons are
colored orange with inhibitor 1a carbons colored green and GSH carbons colored cyan. Hydrogen bonds are depicted as yellow dashed lines.
The coordinates have been deposited in the Brookhaven Protein Data Bank (PDB code 6N69). This figure was generated using PyMOL version
1.7.64.6 (The PyMOL Molecular Graphics System, Version 1.7.64 Schrodinger, LLC).
1.2 Chemistry
The quinoline-3-carboxamides were prepared as depicted in Scheme 1. First, anilines 2a-2b were thermally
condensed with diethyl 2-(ethoxymethylene)malonate 3 via Michael addition and ethanol β-elimination to afford the
phenylaminomethylenemalonates 4a-4b.33 Subsequent phosphorus oxychloride catalyzed Friedel-Craft cyclic
acylation of the phenylaminomethylenemalonates 4a-4b provided the 4-chloroquinolines 5a-5b. Then, the 4-chloro
substituents were removed via palladium-catalyzed hydrogenation to give the quinoline esters 6a-6b.34 Hydrolysis

of esters 6a-6b with metal hydroxides afforded the 3-quinoline acids 7c-7d. Commercially available 3-quinoline
carboxylic acids 7a-7b, as well as 3-quinoline acids 7c-7d, were coupled with various commercially available
amines 8a-8ar, 8au-8bd, and 8bh-8bk or amines 8as-8at and 8be-8bg, known in the chemical literature, to provide
the desired 3-quinoline carboxamides 1d-1bv.
Scheme 1. Synthesis of 3-quinoline carboxamides 1d-1bv.
O
O
O
O
O
R1
N
O
+
R1
NH2
H
R2
R2
O
O
O
3
a
2a R1 = OCF3, R2 = H
2b R1 = OCHF2, R2 = H
4a R1 = OCF3, R2 = H
4b R1 = OCHF2, R2 = H
Cl
O
O
O
O
R1
N
R1
N
b
c
d
R2
R2
5a R1 = OCF3, R2 = H
5b R1 = OCHF2, R2 = H
6a R1 = OCF3, R2 = H
6b R1 = OCHF2, R2 = H
O
O
R3
OH
+
N
R3
H
H2N
R1
N
R1
N
e
R2
R2
7a R1 = OMe, R2 = H
7b R1 = H, R2 = OMe
7c R1 = OCF3, R2 = H
7d R1 = OCHF2, R2 = H
8a - 8bk
1d - 1bv
Reagents and conditions: a) PhMe, 120 °C or EtOH, 65 °C, 99%; b) POCl3, sealed tube, 125-150 °C, 31-33%; c)
Pd(PPh3)2Cl2, Et3SiH, MeCN, 80 °C or H2/Pd-C, EtOH, 93-96%; d) LiOH or NaOH, THF or MeOH, H2O, 50 °C,
56-96%; e) T3P® or HATU, iPr2NEt, CH2Cl2 or THF or DMF, 7-95%.
1.3 Results and discussion
Exploration of the 3-position began with simple aliphatic amines. The primary carboxamide 1f (IC50 = 8,900 nM)
was a slightly less active inhibitor of H-PGDS than its N-methyl congener 1d, while the tertiary
dimethylcarboxamide 1g (IC50 = >50,000 nM) had no measurable inhibitory activity. Subsequent co-crystal
structures revealed that the carboxamide NH forms a hydrogen bond with the cysteine sulfur of the glutathione
cofactor (vide infra). Loss of this intramolecular interaction and occupation of the glutathione cofactor binding
pocket likely accounts for this large loss of inhibitory activity. Both the ethyl and isopropyl carboxamides 1h (IC50
= 1,600 nM) and 1i (IC50 = 870 nM) are more potent that the methyl congener 1d, while the tert-butyl carboxamide
1j (IC50 = 2,400 nM) has similar potency. Thus, analysis of the initial structure activity relationships from the
amide coupling array chemistry revealed that disubstitution at the α-carbon of the amine enhanced potency and a
free NH moiety was crucial for inhibition.
Since the isopropyl carboxamide 1i had submicromolar inhibitory activity, a series of heteroaryl carboxamides were
prepared, following Scheme 1, with the goals of establishing additional π-stacking interactions and/or hydrogen
bonds with the protein or glutathione cofactor and potentially enhancing water solubility relative to the hydrophobic
alkyl derivatives. All of the synthesized heteroaryl analogs enhanced inhibitory activity, with the thiazole 1k (IC50
49 nM) and pyrazoles 1o (IC50 = 57 nM) and 1n (IC50 = 200 nM) being more active than the imidazole 1l (IC50
=
=
390 nM). The differences in activity of these analogs likely results from the energetic differences of their relative
desolvation. Furthermore, substitution adjacent to the amine functionality was detrimental to H-PGDS potency
(compare 1m (IC50 = 8,500 nM) to 1l or 1q (IC50 = 220 nM) to 1o), while more distal substitution was tolerated
(compare 1p (IC50 = 78 nM) and 1r (IC50 = 73 nM) to 1o), with larger groups leading to more potent analogs
(compare 1s (IC50 = 23 nM) and 1t (IC50 = 17 nM) to 1o), albeit with decreases in ligand efficiency.

Heteroaryl rings extended from the amino group by a single atom linker were also synthesized following Scheme 1
to further explore potential protein/cofactor binding interactions. Although the benzyl derivative 1u (IC50 = 71 nM)
exhibited similar activity to the shorter thiazole 1k, all compounds with increased hydrophilic heteroaryl analogs,
including the pyrazoles 1v (IC50 = 320 nM), 1w (IC50 = 160 nM), and 1x (IC50 = 360 nM), imidazole 1y (IC50 = 530
nM), triazole 1z (IC50 = 4,500 nM), tetrazole 1aa (IC50 = 420 nM), pyrazines 1ab (IC50 = 1,400 nM) and 1ae (IC50
2,300 nM), pyrimidines 1ac (IC50 = 1,400 nM), 1ad (IC50 = 1,400 nM), and 1af (IC50 = 2,000 nM), and 2-pyridone
1ag (IC50 = 250 nM) were less potent H-PGDS inhibitors than the thiazole 1k. Furthermore, ring substituents
(compare 1v to 1w and 1x) and methylene linker substituents (compare 1ab to 1ae and 1ac to 1af) in this benzylic
series provided no increased inhibitory activity.
=
As shown in Table 1, the saturated cyclopentyl and cyclohexyl analogs 1ah (IC50 = 200 nM) and 1ai (IC50 = 100
nM) were more potent inhibitors than the corresponding isopropyl derivative 1i. Saturated rings with their sp3
character can disrupt crystal packing density, potentially enhancing dissolution. To explore this concept, an array of
aliphatic analogs with potential hydrogen bond donor/acceptor moieties were prepared, hoping to maintain and/or
improve inhibitory activity while enhancing solubility. Many of the saturated ring analogs, such as the
tetrahydropyran 1aj (IC50 = 170 nM), the cyclohexanol 1ak (IC50 = 110 nM), the piperidine amide 1al (IC50 = 250
nM), the piperidine sulfonamide 1am (IC50 = 110 nM), and the δ-lactam 1an (IC50 = 220 nM), exhibited similar
potencies to their saturated counterparts 1ah and 1ai. Potency was maintained, despite higher desolvation energies
for these compounds and indicated that an additional protein interaction might be established that compensates for
the desolvation cost. In contrast, the less constrained analogs, such as the ethyleneurea 1ao (IC50 = 520 nM), the
glycinamide 1ap (IC50 = 6,900 nM), and the alaninamides 1aq (IC50 = 1,800 nM) and 1ar (IC50 = 7,800 nM), were
poor inhibitors of the enzyme.
As a follow up to these results, the tetrahydrofuran 1as (IC50 = 590 nM) was prepared and it was found to be slightly
less active that the tetrahydropyran 1aj. In contrast, the γ-lactam had similar potency to the δ-lactam 1an, with most
of the activity residing in the (S)-enantiomer 1at (IC50 = 98 nM), rather than the (R)-isomer 1au (IC50 = 15,000 nM).
Substituted lactams, such as the N-methyl analog 1av (IC50 = 1,400 nM), the 4-hydroxy analog 1ax (IC50 = 640 nM),
and the 3-methyl analog 1ay (IC50 = 4,100 nM), as well as the isoxazolidine 1aw (IC50 = 630 nM) and the indoline
1az (IC50 = 350 nM), had less inhibitory activity. In contrast, the more lipophilic indene 1ba (IC50 = 16 nM) was
20-fold more potent that the indoline 1az, likely because of a reduced energetic desolvation penalty upon binding to
the enzyme. The location of the lactam carbonyl was important for maximum inhibition as the isomeric γ-lactams
1bb (IC50 = 1,200 nM) and 1bc (IC50 = 620 nM) were less active that 1at.
The unsubstituted piperidine 1bd (IC50 = 2,200 nM) was 20-fold less potent than the piperidine sulfonamide 1am.
The decrease in inhibitory potency may be the result of either the overt positive charge and/or the significantly
higher desolvation cost on binding. Despite having a high polar surface area with accompanying high desolvation,
the piperidine tetrazole 1be (IC50 = 56 nM) is 4-fold more potent than the piperidine amide 1al. Furthermore, the
piperidine thiazole 1bf (IC50 = 9.7 nM) is an even better H-PGDS inhibitor than 1be. The lower polar surface (67
Å2 versus 98 Å2) likely explains this increase in inhibition.
Other cyclohexyl derivatives, prepared to follow up on the cyclohexanol analog 1ak, also exhibited good H-PGDS
inhibitory activity, with the tertiary alcohols 1bg (IC50 = 25 nM) and 1bk (IC50 = 26 nM), as well as the amide 1bh
(IC50 = 57 nM), being slightly more potent than the tertiary alcohol 1bi (IC50 = 120 nM). Furthermore, the
spiro[3.3]heptane cyclohexyl mimic 1bj (IC50 = 8.2 nM) is an even more potent inhibitor than 1bg. Interestingly,
the amine analog 1bl (IC50 = 14 nM), in which the basicity of the amine has been attenuated via the electron
withdrawing ability of the 2,2,2-trifluoroethyl group, also exhibits excellent inhibitory activity, as compared to the
more basic piperidine analog 1bd.
Since the spiro[3.3]heptane analog 1bj was an extremely potent inhibitor, several four-membered ring analogs were
prepared with the aim of reducing the molecular weight and lipophilicity, while improving water solubility and
maintaining potency. These analogs were less active inhibitors than the parent cyclohexanol 1ak, with the cis
alcohols 1bm (IC50 = 340 nM) and 1bo (IC50 = 280 nM) being more active than the trans congeners 1bn (IC50 = 500
nM) and 1bp (IC50 = 610 nM).
Table 1. H-PGDS enzyme inhibition data.

R1
R2
N
R3
#
R1
R2
R3
IC50 ±
S.D.a
nM
#
R1
R2
R3 IC50 ±
S.D.a
nM
O
1a
CN
CN
CN
H
OCH3
H
H
H
>50000 1al
OCH3
OCH3
OCH3
H
H
H
250
±110
O
HN
O
N
N
1b
1c
13000 1am
±7800
110
±43
O
S
O
HN
OCH3
8400
1an
220
O
NH
N
±3100
±110
N
H
O
O
O
O
O
O
O
1d
1e
1f
OCH3
H
H
OCH3
H
3100
±1700
1ao
1ap
1aq
OCH3
OCH3
OCH3
OCH3
H
H
H
H
520
±220
NH
NH
NH
NH2
N
N
H
O
NH2
7700
±2500
6900
±8700
O
O
O
NH
NH
NH
O
NH2
OCH3
OCH3
8900
±2300
1800
±960
O
NH2
1g
H
>50000 1ar
7800
±2400
O
O
O
O
1h
1i
OCH3
OCH3
H
H
1600
±240
1as
1at
OCH3
OCH3
H
H
590
±160
NH
N
H
870
±350
98
±42
O
NH
NH
NH
N
H
O
O
O
O
1j
OCH3
H
2400
±480
1au
OCH3
H
15000
±8800
O
O
NH
N
H
O
N
S
1k
1l
OCH3
OCH3
H
H
49
±37
1av
OCH3
OCH3
H
H
1400
±1100
N
N
N
H
N
H
O
390b
±25
1aw
630
±85
O
O
HN
O
NH
N
H
N
H
O
HO
1m
OCH3
H
8500c 1ax
±5500
OCH3
H
640
±310
N
O
O
NH
O
N
N
N
H
N
H
O
O
1n
1o
OCH3
OCH3
H
H
200
±39
1ay
1az
OCH3
OCH3
H
H
4100
±2300
NH
NH
N
H
N
H
O
O
57
±27
350
±110
NH
NH
N
O
O
N
N
N
H
N
H
O
1p
OCH3
H
78
±35
1ba
OCH3
H
16
±6.5
N
H
N
H

NH
NH
O
O
O
1q
1r
1s
1t
OCH3
OCH3
OCH3
OCH3
H
H
H
H
220
±58
1bb
1bc
1bd
1be
OCH3
OCH3
OCH3
OCH3
H
H
H
H
1200
±740
N
O
O
N
N
H
N
H
NH
NH
NH
O
O
O
N
N
N
73
±11
620
±290
N
H
N
H
23
±12
2200
±480
O
NH
N
H
N
H
O
17
±5
56
±64
N
N
N
N
N
H
HN
N
N
O
HN
O
1u
1v
1w
OCH3
OCH3
OCH3
H
H
H
71
±54
1bf
1bg
1bh
OCH3
OCH3
OCH3
H
H
H
9.7
±3.9
O
O
O
O
O
O
S
N
NH
NH
NH
320
±280
25
±3.4
N
N
N
H
HN
OH
O
O
160
±130
57
±23
N
N
HN
HN
O
HN
O
1x
1y
OCH3
OCH3
H
H
360
±320
1bi
1bj
OCH3
OCH3
H
H
120
±12
N
N
NH
NH
NH
OH
530
±140
8.2
±2.4
N
N
HN
OH
OH
O
1z
OCH3
OCH3
OCH3
H
H
H
4500
±3300
1bk
1bl
OCH3
OCH3
OCH3
H
H
H
26
±5.3
N
N
H
HN
O
O
F
F
N
1aa
1ab
420
±210
14
±2.2
N
N
O
F
N
H
NH
HN
NH
N
O
HN
O
1400
±920
1bm
340
±190
O
O
OH
NH
NH
N
N
1ac
1ad
1ae
1af
OCH3
OCH3
OCH3
OCH3
H
H
H
H
1400
±760
1bn
1bo
1bp
1bq
OCH3
OCH3
OCH3
OCF3
H
H
H
H
500
±230
N
HN
O
OH
N
1400
±710
280
±27
O
NH
N
N
HN
O
OH
OH
2300
±1100
610
±98
O
O
NH
NH
N
N
HN
O
2000
±900
230
±48
N
NH
N
H
O
O
1ag
1ah
OCH3
OCH3
H
H
250
±230
1br
1bs
OCF3
OCF3
H
H
120
±28
O
NH
O
N
N
N
N
NH
HN
O
N
O
200
±92
15
±5.2
N
H
HN
OH

O
1ai
1aj
1ak
OCH3
OCH3
OCH3
H
H
H
100
±61
1bt
1bu
1bv
OCHF2
OCHF2
OCHF2
H
H
H
52
±13
O
O
NH
N
N
H
H
O
O
170
±81
20
±6.8
O
N
N
N
N
HN
O
N
N
H
O
110
±44
9.9
±2.7
HN
OH
HN
OH
aS.D. = standard deviation; mean ± S.D. All inhibitors were tested with N ≥ 4 unless overwise noted.
bN = 2.
cN = 3.
Many of these compounds had H-PGDS inhibitory potencies less than 100 nM in the enzymatic assay. Some also
had reduced plasma protein binding with free fractions of greater than 5%, resulting in small potency decreases in a
rat basophilic leukemia (RBL) cell PGD2 production assay relative to the H-PGDS enzymatic assay. Despite their
good ex vivo H-PGDS inhibitory potency, these analogs all contained a 3-carboxamide moiety. Carboxamides are
susceptible to hydrolysis by amidases in vivo and some of these analogs were enzymatically degraded to the
corresponding carboxylic acid in a whole blood assay (e.g. human whole blood assay % remaining after 1 hour: 1j
= 1%, 1n = 63%). Substitution was the key to amidase recognition with certain substituents presumably being poor
substrates of the putative amidase and having increased plasma stability (e.g. human whole blood assay % remaining
after 1 hour: 1o = 93%, 1at = 100%).
These analogs also all contained a 7-methoxyquinoline moiety. These 7-methoxyquinoline-3-carboxamides are
quite electron rich, very UV active, and could exhibit phototoxicity. In fact, some of the analogs degrade over time
when exposed to UV light. Furthermore, aryl methoxy groups are a potential metabolic liability due to their
potential for oxidative O-dealkylation of the methyl moiety to produce quinolin-7-ols. The phenolic metabolites
may have reduced H-PGDS inhibitory activity and are susceptible to secondary phase metabolism with the resulting
conjugates having increased renal clearances. Therefore, the stability of a representative analog 1bg was studied in
a mouse liver microsome assay and it exhibited at short in vitro half-life (t1/2 = 38 minutes).
Hypothesizing that this metabolic instability arose from O-methyl dealkylation, analogs of 1at, 1be, and 1bg were
prepared that replaced the methyl moiety with more electron deficient difluoromethyl or trifluoromethyl in attempts
to increase the metabolic stability of these H-PGDS inhibitors. These electron withdrawing substitutions also
attenuate the UV absorbance of these isomers, enhancing their photostability. The trifluoromethyl derivatives 1bq
(IC50 = 230 nM), 1br (IC50 = 120 nM), and 1bs (IC50 = 15 nM) tended to be less potent inhibitors of H-PGDS than
their difluoromethyl analogs 1bt (IC50 = 52 nM), 1bu (IC50 = 20 nM), and 1bv (IC50 = 9.9 nM). The best two of
these competitive, reversible, potent analogs, 1bs (RBL IC50 = 100 nM) and 1bv (RBL IC50 = 100 nM), also
exhibited good cellular activity in the rat basophilic leukemia (RBL) cell PGD2 production assay and were
substantially more stable in mouse liver microsomes, both having in vitro half-lives of greater than 180 minutes,
supporting the O-demethylation hypothesis.
Furthermore, as shown in Table 2, both 1bs (PAPP = 470 nm/sec) and 1bv (PAPP = 240 nm/sec) were highly
permeable in an artificial membrane permeability assay that served as a surrogate for the more labor intensive
Madin-Darby canine kidney (MDCK) cell permeation assay.35 Thus, the oral bioavailability of these analogs should
not be limited by absorption across cell membranes. Additionally, the fasted state simulated intestinal fluid
solubility’s of 1bs (FaS-SIF = 0.016 mg/mL) and 1bv (FaS-SIF = 0.070 mg/mL) were modest, but due to their high
permeability, acceptable for i.v. and p.o. pharmacokinetic profiling.36
With the view of identifying a suitable tool compound to explore the pharmacodynamics of H-PGDS inhibitors in
murine models of inflammatory diseases, the pharmacokinetic parameters of these compounds were determined in
mice. The trifluoromethyl analog 1bs had a low i.v. clearance (Cl = 3.8 mL/min/kg), moderate steady state volume
of distribution (VSS = 2.5 L/kg), and long terminal half-life (t1/2 = 7.9 h). The compound also exhibited good oral
exposure (p.o. DNAUC = 1900 ng/hr/mL) with a moderate oral bioavailability (F = 46%). The difluoromethyl

analog 1bv also had a low i.v. clearance (Cl = 5.6 mL/min/kg), moderate steady state volume of distribution (VSS
=
1.7 L/kg), and long terminal half-life (t1/2 = 3.8 h), as well as good oral exposure (p.o. DNAUC = 1800 ng/hr/mL)
with a higher oral bioavailability (F = 61%).
Table 2. H-PGDS inhibitor pharmacokinetic data
#
PAPP
FaS-SIF
Microsomes
t1/2
Cl
VSS
F
DNAUC
nm/sec
470
240
mg/mL
0.016
0.070
t1/2 min
>180
>180
h
7.9
3.8
mL/min/kg
L/kg
2.5
1.7
%
46
61
ng/hr/mL
1900
1bs
1bv
3.8
5.6
1800
To further explore the suitability of compound 1bv for in vivo studies, its binding kinetics were determined. The
dissociation constant Kd for 1bv (Kd = 0.50 ± 0.08 nM) was measured in the presence of glutathione (1 mM) by
equilibrium titration of the quenching of the indole intrinsic fluorescence of 104Trp in the active site with 1bv excited
at 285 nm. The kon of 1bv (kon = 5.4 ± 0.19 X 106 M-1sec-1) was measured by stopped-flow spectrofluorimetry from
the fastest phase of intrinsic tryptophan quench time courses, while the koff (calculated koff = 0.0027 sec-1) was
calculated from the values of Kd and kon (Kd = koff/kon). Inhibitor 1bv rapidly associates with and dissociates from
H-PGDS. This Kd is a better reflection of the intrinsic potency of 1bv as compared to the IC50 of 1bv, which is
elevated due to prostaglandin substrate competition and active site titration of hH-PGDS in the activity assay.
Several other enzymes also employ PGH2 as a substrate to produce prostanoids. Their inhibition by an H-PGDS
inhibitor tool compound could confound the interpretation of pharmacodynamic studies in inflammatory models of
disease, therefore the selectivity of inhibitor 1bv versus two closely related synthase enzymes in the prostanoid
pathway was also determined. Analog 1bv was tested against lipocalin prostaglandin D synthase (L-PGDS, vide
supra)15 and microsomal prostaglandin E synthase (m-PGES)37 and did not inhibit either enzyme at concentrations
up to 30,000 nM, supporting its suitability for use in vivo to probe the effects of H-PGDS inhibition.
Furthermore, the general off-target liabilities of 1bv were also evaluated. Compound 1bv did not block the human
ether-a-go-go-related gene potassium ion channel (Kv11.1)38 or inhibit cytochrome p450 3A4 monooxygenase
enzyme at concentrations up to 30,000 nM. Also, at concentrations up to 10,000 nM, 1bv did not inhibit a broad
panel of proteins, including 7-TM receptors (5HT1B, 5HT2A, 5HT2C, α2C, β2, κ, μ, A2A, CB2, D1, D2, H1, M1, M2,
NK1, V1A), ion channels (5HT3, CaV1.2, GABAA, KV7.1, NaV1.5, NMDA), enzymes (AChE, COX-2, MAO-B,
PDE3A, PDE4B), transcription factors (AhR, AR, PXR), kinases (Aurora B, LCK, PI3Kγ), and transporters (NET,
OATP1B1, SERT). Coupled with its high H-PGDS inhibitory potency and good pharmacokinetic characteristics, this
profile makes 1bv a suitable tool compound for studying the pharmacodynamic effects of H-PGDS inhibition.
Inhibitor 1bv was profiled in a murine mast cell degranulation model of inflammation to evaluate the acute effects
of a H-PGDS inhibitor on mast cell release of PGD2.39 Male C57BL/6J mice were administered p.o. with vehicle or
H-PGDS inhibitor 1bv at various doses (0.03, 0.1, 0.3, 1, 3 mg/kg), then anesthetized and challenged one hour later
with vehicle (PBS) or the N-methyl-p-methoxyphenethylamine/formaldehyde synthetic polymer 48/80 (i.p.0.75
mg/mL) with abdomen massage. After seven minutes, blood was collected (1bv analysis), the mice were
euthanized, and lavage fluid was collected from the abdominal cavity (PGD2 analysis). As shown in Figure 3, the
mast cell secretagogue 48/80 induces production and release of PGD2 into peritoneal lavage fluid versus phosphate
buffered saline (PBS). H-PGDS inhibitor 1bv attenuates this PGD2 release to baseline levels in a dose-dependent
manner with an ED50 = 0.032 mg/kg (1bv blood EC50 = 21 nM) in this acute inflammation model. Levels of the
related prostaglandin PGE2 were not significantly different between 1bv treated and untreated animals after
challenge with synthetic polymer 48/80.
Figure 3. H-PGDS inhibition blocks 48/80 challenge-induced PGD2 production in vivo

400
300
200
100
0
*
*
*
*
0
0
0
0
0
0
8
S
8
8
8
8
/
/8
/
/
/
/
8
8
8
8
8
8
PB
4
4
4
4
4
4
+
+
+
+
+
+
+
h
h
Ve
bv
1
bv
bv
bv
bv
1
Ve
1
1
1
g
g
g
g
g
k
*P<0.05 vs. Veh+48/80
ED50=0.032 mg/kg
k
k
k
mg/k
mg/
mg/
mg/
mg/
0
3
3
0
.1
.
.
.
.0
0
0
1
3
0
Figure 3 Legend: Young adult male C57BL/6J mice were administered (p.o.) vehicle or the H-PGDS inhibitor 1bv at varying doses (from 0.03
mg/kg to 3 mg/kg). One hour later, mice were anesthetized, injected with (i.p.) 0.2 mL vehicle or 48/80 (0.75 mg/mL), and peritoneal lavage
fluid was collected after 7 minutes. (A) Data are presented as means (n = 7/group) ± S.E.M. and analyzed using ANOVA followed by Dunnett’s
comparing vehicle+48/80 to respective treatment group.
To gain further understanding of the binding mode of quinoline H-PGDS inhibitors, a protein co-crystal structure of
inhibitor 1bv bound with hH-PGDS and GSH was obtained. As shown in Figure 4, the bound structure of 1bv has
many similarities to that of fragment 1a. Both utilize the quinoline nitrogen to form a key hydrogen bond with a
tightly bound water molecule (structural water oxygen to quinoline nitrogen distance = 2.8 Å) coordinated by
199Leu, 159Thr, and an additional water molecule (vide supra). Both compounds also form a face to face π-π stacking
interaction between the quinoline ring and the indole side chain of 104Trp (face of indole 104Trp to face of quinoline
1bv distance = 3.6 Å). In addition, the quinoline 3-carboxamide NH forms a hydrogen bond with the sulfur of the
glutathione cofactor cysteine residue. This interaction likely accounts for the 4-fold gain in potency between the
nitrile 1b and the N-methyl carboxamide 1d. Also, the difluoromethoxy group extends into a small hydrophobic
pocket formed by 99Met, 155Ile, and 13Gly. Additionally, the electron releasing character of the difluoromethoxy
group leads to increased basicity of the quinoline nitrogen that results in an enhanced polar interaction between that
nitrogen and the coordinating water molecule. The combination of enhanced polar and hydrophobic contacts could
explain the 10-fold increase in inhibitory activity of 1b versus the starting fragment 1a. Finally, the chair
conformation of the cyclohexyl moiety enhances binding through hydrophobic interactions with the side chains of
105Ala and 9Phe. These interactions help explain the ~500-fold gain in activity of 1bg versus 1d. The tertiary
alcohol is mostly solvent exposed, adding little to ligand binding, but enhancing water solubility of this analog.
Thus, these human H-PGDS co-crystal structures bound with ligand and cofactor not only provide structural insight
into the binding requirements of the enzyme, but also help to rationalize much of the SAR observed in the quinoline
carboxamide inhibitor series.
Figure 4. Co-crystal structure of inhibitor 1bv bound to hH-PGDS with GSH.

Figure 4 Legend. Ligand binding site of the X-ray co-crystal structure of 1bv complexed with hH-PGDS and glutathione (GSH). The hH-
PGDS carbons are colored green with inhibitor 1bv carbons colored magenta and GSH carbons colored cyan. Hydrogen bonds are depicted as
yellow dashed lines. The coordinates have been deposited in the Brookhaven Protein Data Bank (PDB code 6N4E). This figure was generated
using PyMOL version 1.7.64.6 (The PyMOL Molecular Graphics System, Version 1.7.64 Schrodinger, LLC).
1.4 Conclusion
In this work, further evidence of the utility of fragment-based drug discovery to deliver optimizable chemical matter
for robust lead development is provided. A diverse set of confirmed hits, derived from ligand-bound protein crystal
structures of H-PGDS, were clustered by binding mode and resulted in the identification of a cluster containing the
6,6-fused heterocyclic quinoline 1a. Through SAR by catalog, array chemistry, traditional medicinal chemistry, and
structure-based design, this initial hit was developed into a H-PGDS chemical tool compound, GSK2894631A 1bv,
with excellent potency, selectivity, and similar ligand efficiency (LE = 0.42) to the starting fragment 1a.
Furthermore, 1bv exhibited good pharmacokinetic parameters in mice and dose-dependently attenuated PGD2
production and secretion in a mast cell degranulation assay pharmacodynamic model of inflammation. Further
publications from these laboratories will explore its use in other prostaglandin-driven inflammation models.
1.5 Experimental section
All commercial chemicals and solvents were reagent grade and were used without further purification unless otherwise
specified. The following abbreviations are utilized in the manuscript: tetrahydrofuran (THF), diethyl ether (Et2O),
dimethyl sulfoxide (DMSO), ethyl acetate (EtOAc), dichloromethane (CH2Cl2), trifluoroacetic acid (TFA), N,N-
dimethylformamide (DMF), methanol (MeOH), dimethoxyethane (DME), N-methylpyrrolidine (NMP), acetonitrile
(MeCN), chloroform (CHCl3), phosphorus oxychloride (POCl3), magnesium sulfate (MgSO4), triethylamine (Et3N),
2-propanol (iPrOH), diisopropylethylamine (iPr2NEt), sodium hydroxide (NaOH), t-butylmethyl ether (TBME),
acetic acid (AcOH or HOAc), ethanol (EtOH), di-tert-butyldicarbonate (BOC2O), sodium sulfate (Na2SO4), N,N-
dimethylacetamide (DMA), sodium bicarbonate (NaHCO3), potassium carbonate (K2CO3), 1-[bis(dimethylamino)-
methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (HATU), azobis(isobutyronitrile)
(AIBN), 4-(2-hydroxyethyl)-piperazine-1-ethanesulfonic acid (HEPES), and dithiothreitol (DTT). All reactions
except those in aqueous media were carried out with the use of standard techniques for the exclusion of moisture.
Reactions were monitored by thin-layer chromatography (TLC) on 0.25 mm silica gel plates (60F-254, E. Merck) and
visualized with UV light, iodine, iodoplatinate, potassium permanganate, cerium molybdate, or 5% phosphomolybdic
acid in 95% ethanol. Final compounds were typically purified either by flash chromatography on silica gel (E. Merck
40−63 mm), radial chromatography on a Chromatotron™ using prepared silica gel plates, or on a Biotage Horizon or
ISCO Combiflash® pump and fraction collection system utilizing prepacked silica gel. Analytical purity was assessed
either by reversed-phase high performance liquid chromatography (RP-HPLC) using an Agilent 1100 system equipped
with a diode array spectrometer (λ range 190−400 nm) or by the LC-MS method detailed below. The stationary phase
was a Keystone Scientific BDS Hypersil™ C-18 column (5 μm, 4.6 mm × 200 mm). The mobile phase employed
0.1% aqueous TFA with MeCN as the organic modifier and a flow rate of 1.0 mL/min. Analytical data are reported
as retention time (tR) in minutes and percent purity. All compounds were found to be ≥95% pure unless otherwise
1
indicated. H NMR spectra were recorded on either a Varian Unityplus™ 400 MHz or a Bruker Avance™ III 400
MHz NMR spectrometer. Chemical shifts are reported in parts per million (ppm, δ units). Coupling constants are
reported in units of hertz (Hz). Splitting patterns are designated as s, singlet; d, doublet; t, triplet; q, quartet; p, pentet;
h, hextet; m, multiplet; or br, broad. Low-resolution mass spectra (MS) were recorded on a Waters SQD. High-
resolution MS were recorded on a Waters (Micromass®) LCT time-of-flight mass spectrometer. Low-resolution mass
spectra were obtained under electrospray ionization (ESI), atmospheric pressure chemical ionization (APCI), or fast
atom bombardment (FAB) methods. All studies were conducted in accordance with the GSK Policy on the Care,
Welfare and Treatment of Laboratory Animals and were reviewed the Institutional Animal Care and Use Committee
either at GSK or by the ethical review process at the institution where the work was performed.
General amide bond coupling method A. N,N-Diisopropylethylamine (4.0 eq) was added to acid (1 eq) in solvent
(dichloromethane, tetrahydrofuran, or N,N-dimethylformamide, 0.05 to 0.2 M) at room temperature. Then, amine
(1.0-2.0 eq) was added and the reaction mixture was stirred for five minutes. Then, n-propylphosphonic acid
anhydride (2.0 eq) was added and the reaction mixture was stirred for sixteen hours. The reaction mixture was
concentrated. The resulting residue was purified by RP HPLC or silica gel chromatography to give the quinoline-3-
carboxamide (25%-86 % yield).

General amide bond coupling method B. N,N-Diisopropylethylamine (3.0–4.0 eq) was added to 8-
methoxyquinoline-3-carboxylic acid (1.0 eq) in solvent (dichloromethane, tetrahydrofuran, or N,N-
dimethylformamide, 0.05 to 0.2 M) at room temperature. Then, 1-((dimethylamino)(dimethyliminio)methyl)-1H-
[1,2,3]triazolo[4,5-b]pyridine 3-oxide hexafluorophosphate(V) (1.0-1.5 eq) was added and the reaction mixture was
stirred for five minutes. Then, amine (1.0 – 2.0 eq) was added and the reaction mixture was stirred for one to sixteen
hours. 10% Aqueous citric acid was added and the reaction mixture was extracted with dichloromethane, washed
with saturated sodium bicarbonate, dried over magnesium sulfate, filtered, and concentrated. The resulting residue
was purified by RP HPLC or silica gel chromatography to give the quinoline-3-carboxamide (1%-95 % yield).
Quinoline-3-carbonitrile 1a.
N
N
1a
Commercial (Aldrich); 1H NMR (400 MHz, CD3SOCD3) δ 9.17 (d, 1H, J = 2 Hz), 9.09 (d, 1H, J = 2 Hz), 8.13 (d, 1H,
J = 8 Hz), 8.11 (d, 1H, J = 8 Hz), 7.98 (ddd, 1H, J = 8, 7, 2 Hz), 7.78 (ddd, 1H, J = 8, 7, 1 Hz); LC-MS (LC-ES) M+H
= 155.
7-Methoxyquinoline-3-carbonitrile 1b.
N
O
N
1b
Commercial (Biofine); 1H NMR (400 MHz, CD3SOCD3) δ 9.08 (d, 1H, J = 2 Hz), 8.94 (d, 1H, J = 2 Hz), 8.00 (d, 1H,
J = 9 Hz), 7.48 (d, 1H, J = 2 Hz), 7.41 (dd, 1H, J = 9, 2 Hz), 3.97 (s, 3H); LC-MS (LC-ES) M+H = 185.
8-Methoxyquinoline-3-carbonitrile 1c.
N
N
O
1c
Commercial (Astatech); 1H NMR (400 MHz, CD3SOCD3) δ 9.09 (d, 1H, J = 2 Hz), 9.03.76 (d, 1H, J = 2 Hz), 7.69 (t,
1H, J = 8 Hz), 7.61 (dd, 1H, J = 8, 1 Hz), 7.41 (dd, 1H, J = 8, 1 Hz), 3.99 (s, 3H); LC-MS (LC-ES) M+H = 185.
7-Methoxy-N-methylquinoline-3-carboxamide 1d.
O
N
H
O
N
1d
Commercial (Aurora Building Blocks); Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); Methyl
1
amine 8a (Aldrich); N,N-Dimethylformamide; 46% yield; H NMR (400 MHz, CDCl3) δ 9.19 (d, 1H, J = 2 Hz), 8.60
(d, 1H, J = 2 Hz), 7.78 (d, 1H, J = 8 Hz), 7.45 (d, 1H, J = 2 Hz), 7.26 (dd, 1H, J = 8, 2 Hz), 6.76 (br s, 1H), 3.98 (s,
3H), 3.08 (d, 3H, J = 5 Hz); LC-MS (LC-ES) M+H = 217.
8-Methoxy-N-methylquinoline-3-carboxamide 1e.

O
N
H
N
O
1e
Commercial (Aurora Building Blocks); Method A; 8-Methoxyquinoline-3-carboxylic acid 7b (Astatech);
Methylamine 8a (Aldrich); Dichloromethane; 86% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.18 (d, 1H, J = 2 Hz),
8.80 (br q, 1H, J = 5 Hz), 8.72 (d, 1H, J = 2 Hz), 7.59 (d, 1H, J = 5 Hz), 7.58 (d, 1H, J = 5 Hz), 7.28 (t, 1H, J = 5 Hz),
3.97 (s, 3H), 2.84 (d, 3H, J = 5 Hz); LC-MS (LC-ES) M+H = 217.
7-Methoxyquinoline-3-carboxamide 1f.
O
NH2
O
N
1f
Commercial (ABC Lab); 19 M Sodium hydroxide (0.057 ml, 1.088 mmol) was added to 7-methoxyquinoline-3-
carbonitrile 1b (0.0668 g, 0.363 mmol) in dimethyl sulfoxide (1.813 ml) at room temperature, followed by 30%
aqueous hydrogen peroxide (0.381 ml, 4.35 mmol) and the solution was stirred for sixty-four hours at room
temperature. Water was added to the reaction mixture and it was extracted with dichloromethane (4X), dried over
magnesium sulfate, filtered, and concentrated. The residue was purified by silica gel chromatography, eluting with
methanol:ethyl acetate (0:1 to 2:3) to give 7-methoxyquinoline-3-carboxamide 1f (Commercial (ABCR), 0.0352 g,
1
0.165 mmol, 45.6 % yield). H NMR (400 MHz, CD3SOCD3) δ 9.22 (d, 1H, J = 2 Hz), 8.75 (d, 1H, J = 2 Hz), 8.22
(br s, 1H), 7.96 (d, 1H, J = 9 Hz), 7.59 (br s, 1H), 7.44 (d, 1H, J = 2 Hz), 7.31 (dd, 1H, J = 9, 2 Hz), 3.94 (s, 3H);
LC-MS (LC-ES) M+H = 203.
7-Methoxy-N,N-dimethylquinoline-3-carboxamide 1g.
O
N
O
N
1g
Commercial (Aurora Building Blocks); Method A; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech);
1
Dimethylamine 8b (Aldrich); Dichloromethane; 73% yield; H NMR (400 MHz, CD3SOCD3) δ 8.83 (d, 1H, J = 2
Hz), 8.38 (d, 1H, J = 2 Hz), 7.95 (d, 1H, J = 9 Hz), 7.42 (d, 1H, J = 2 Hz), 7.31 (dd, 1H, J = 9, 2 Hz), 3.93 (s, 3H),
3.02 (s, 6H); LC-MS (LC-ES) M+H = 231.
N-Ethyl-7-methoxyquinoline-3-carboxamide 1h.
O
N
H
O
N
1h
Commercial (Aurora Building Blocks); Method A; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); Ethylamine
8c (Aldrich); Dichloromethane; 74% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.19 (d, 1H, J = 2 Hz), 8.73 (br t, 1H,
J = 6 Hz), 8.71 (d, 1H, J = 2 Hz), 7.98 (d, 1H, J = 9 Hz), 7.44 (d, 1H, J = 2 Hz), 7.31 (dd, 1H, J = 9, 2 Hz), 3.94 (s,
3H), 3.34 (dq, 2H, J = 7, 6Hz), 1.16 (t, 3H, J = 7 Hz); LC-MS (LC-ES) M+H = 231.
N-(1-Methylethyl)-7-(methyloxy)-3-quinolinecarboxamide 1i.

O
N
H
O
N
1i
Commercial (Aurora Building Blocks); Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech);
Isopropylamine 8d (Aldrich); N,N-Dimethylformamide; 42% yield; 1H NMR (400 MHz, CDCl3) δ 9.18 (d, 1H, J = 2
Hz), 8.62 (d, 1H, J = 2 Hz), 8.24 (br s, 1H), 7.80 (d, 1H, J = 8 Hz), 7.51 (d, 1H, J = 2 Hz), 7.27 (dd, 1H, J = 8, 2 Hz),
4.36 (h, 1H, J = 6 Hz), 3.98 (s, 3H), 1.32 (d, 6H, J = 6 Hz); LC-MS (LC-ES) M+H = 217.
N-(tert-Butyl)-7-methoxyquinoline-3-carboxamide 1j.
O
N
H
O
N
1j
Commercial (Aurora Building Blocks); Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); tert-
Butylamine 8e (Aldrich); N,N-Dimethylformamide; 47% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.15 (d, 1H, J =
2 Hz), 8.68 (d, 1H, J = 2 Hz), 8.08 (d, 1H, J = 9 Hz), 8.03 (br s, 1H), 7.97 (d, 1H, J = 2 Hz), 7.31 (dd, 1H, J = 9, 2 Hz),
3.95 (s, 3H), 1.43 (s, 9H); LC-MS (LC-ES) M+H = 259.
7-(Methyloxy)-N-1,3-thiazol-2-yl-3-quinolinecarboxamide 1k.
O
S
N
N
H
O
N
1k
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 2-Aminothiazole 8f (Aldrich); N,N-
Dimethylformamide; 95% yield; 1H NMR (400 MHz, CDCl3) δ 9.50 (d, 1H, J = 2 Hz), 8.87 (d, 1H, J = 2 Hz), 8.21 (s,
1H), 7.88 (d, 1H, J = 9 Hz), 7.52 (d, 1H, J = 2 Hz), 7.45 (d, 1H, J = 3 Hz), 7.32 (dd, 1H, J = 9, 2 Hz), 7.06 (d, 1H, J =
3 Hz), 4.02 (s, 3H); LC-MS (LC-ES) M+H = 286.
N-(1H-Imidazol-2-yl)-7-methoxyquinoline-3-carboxamide 1l.
O
HN
N
N
H
O
N
1l
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 1H-Imidazol-2-amine 8g (Oakwood); N,N-
Dimethylformamide; 81% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.40 (s, 1H), 8.90 (s, 1H), 8.83 (d, 1H, J = 2
Hz), 8.39 (d, 1H, J = 2 Hz), 8.01 (d, 1H, J = 9 Hz), 7.45 (d, 1H, J = 2 Hz), 7.32 (dt, 1H, J = 9, 2 Hz), 6.86 (s, 2H), 3.95
(s, 3H); LC-MS (LC-ES) M+H = 269.
7-Methoxy-N-(1-methyl-1H-imidazol-2-yl)quinoline-3-carboxamide 1m.
O
N
N
N
H
O
N
1m

Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 1-Methyl-1H-imidazol-2-amine 8h (Oakwood);
N,N-Dimethylformamide; 12% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.54 (br s, 1H), 8.90 (s, 1H), 8.02 (d, 1H, J
= 9 Hz), 7.44 (d, 1H, J = 2 Hz), 7.28 (dd, 1H, J = 9, 2 Hz), 7.06 (s, 1H), 6.87 (s, 1H), 3.94 (s, 3H), 3.60 (s, 3H); LC-
MS (LC-ES) M+H = 283.
7-Methoxy-N-(1H-pyrazol-4-yl)quinoline-3-carboxamide 1n.
N
O
NH
N
H
O
N
1n
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 1H-Pyrazol-4-ylamine 8i (Oakwood); N,N-
Dimethylformamide; 79% yield; 1H NMR (600 MHz, CD3SOCD3) δ 12.67 (br s, 1H), 10.67 (s, 1H), 9.31 (d, 1H, J =
2 Hz), 8.83 (d, 1H, J = 2 Hz), 8.02 (d, 1H, J = 9 Hz), 7.89 (br s, 2H), 7.48 (d, 1H, J = 3 Hz), 7.35 (dd, 1H, J = 9, 3
Hz), 3.96 (s, 3H); LC-MS (LC-ES) M+H = 269.
7-Methoxy-N-(1H-pyrazol-3-yl)quinoline-3-carboxamide 1o.
O
NH
N
N
H
O
N
1o
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 3-Aminopyrazole 8j (Oakwood); N,N-
Dimethylformamide; 12% yield; 1H NMR (600 MHz, CD3SOCD3) δ 12.51 (br s,1H), 11.11 (br s, 1H), 9.31 (d, 1H, J
= 2 Hz), 8.92 (d, 1H, J = 2 Hz), 8.00 (d, 1H, J = 9 Hz), 7.78 (br s, 1H), 7.46 (d, 1H, J = 3 Hz), 7.34 (dd, 1H, J = 9, 3
Hz), 6.67 (br s, 1H), 3.95 (s, 3H); LC-MS (LC-ES) M+H = 269.
7-Methoxy-N-(1-methyl-1H-pyrazol-3-yl)quinoline-3-carboxamide 1p.
O
N
N
N
H
O
N
1p
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 1-Methyl-1H-pyrazol-3-ylamine 8k (Oakwood);
N,N-Dimethylformamide; 47% yield; 1H NMR (600 MHz, CD3SOCD3) δ 11.10 (s, 1H), 9.30 (d, 1H, J = 2 Hz), 8.90
(d, 1H, J = 2 Hz), 7.99 (d, 1H, J = 9 Hz), 7.63 (d, 1H, J = 2 Hz), 7.46 (d, 1H, J = 2 Hz), 7.34 (dd, 1H, J = 9, 2 Hz),
6.64 (d, 1H, J = 2 Hz), 3.95 (s, 3H), 3.79 (s, 3H); LC-MS (LC-ES) M+H = 283.
7-Methoxy-N-(1-methyl-1H-pyrazol-5-yl)quinoline-3-carboxamide 1q.
O
N
N
N
H
O
N
1q
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 1-Methyl-1H-pyrazol-4-ylamine 8l (Oakwood);
N,N-Dimethylformamide; 11% yield; 1H NMR (600 MHz, CD3SOCD3) δ 10.62 (br s, 1H), 9.31 (s, 1H), 8.89 (s,
1H), 8.05 (d, 1H, J = 8 Hz), 7.49 (s, 1H), 7.42 (t, 1H, J = 2 Hz), 7.37 (dt, 1H, J = 9, 2 Hz), 6.30 (d, 1H, J = 2 Hz),
3.96 (s, 3H), 3.75 (s, 3H); LC-MS (LC-ES) M+H = 283.
7-Methoxy-N-(5-methyl-1H-pyrazol-3-yl)quinoline-3-carboxamide 1r.

O
NH
N
N
H
O
N
1r
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 3-Amino-5-methylpyrazole 8m (Aldrich); N,N-
Dimethylformamide; 9% yield; 1H NMR (400 MHz, CD3SOCD3) δ 10.90 (br s, 1H), 9.30 (d, 1H, J = 2 Hz), 8.90 (d,
1H, J = 2 Hz), 8.15 (s, 1H), 7.99 (d, 1H, J = 8 Hz), 7.46 (d, 1H, J = 2 Hz), 7.34 (dd, 1H, J = 9, 2 Hz), 6.44 (br s, 1H),
3.96 (s, 3H), 2.24 (s, 3H); LC-MS (LC-ES) M+H = 283.
N-(5-Cyclopropyl-1H-pyrazol-3-yl)-7-methoxyquinoline-3-carboxamide 1s.
O
NH
N
N
H
O
N
1s
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 3-Cyclopropyl-1H-pyrazol-5-amine 8n (Oakwood);
1
N,N-Dimethylformamide; 12% yield; H NMR (400 MHz, CD3SOCD3) δ 10.90 (br s, 1H), 9.32 (d, 1H, J = 2 Hz),
8.94 (d, 1H, J = 2 Hz), 8.13 (s, 1H), 8.02 (d, 1H, J = 8 Hz), 7.47 (d, 1H, J = 2 Hz), 7.37 (dd, 1H, J = 9, 2 Hz), 6.34 (br
s, 1H), 3.97 (s, 3H), 1.96-1.86 (m, 1H), 0.98-0.90 (m, 2H), 0.74-0.68 (m, 2H); LC-MS (LC-ES) M+H = 309.
7-Methoxy-N-(5-phenyl-1H-pyrazol-3-yl)quinoline-3-carboxamide 1t.
O
NH
N
N
H
O
N
1t
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 3-Amino-5-phenylpyrazole 8o (Aldrich); N,N-
Dimethylformamide; 9% yield; 1H NMR (400 MHz, CD3SOCD3) δ 12.93 (br s, 1H), 11.12 (br s, 1H), 9.34 (d, 1H, J
= 2 Hz), 8.95 (d, 1H, J = 2 Hz), 8.02 (d, 1H, J = 8 Hz), 7.78 (d, 2H, J = 7 Hz), 7.48 (d, 1H, J = 2 Hz), 7.47 (t, 2H, J =
7 Hz), 7.36 (t, 1H, J = 7 Hz), 7.35 (dd, 1H, J = 9, 2 Hz), 7.08 (br s, 1H), 3.97 (s, 3H); LC-MS (LC-ES) M+H = 345.
N-Benzyl-7-methoxyquinoline-3-carboxamide 1u.
O
N
H
O
N
1u
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); Benzylamine 8p (Aldrich); N,N-
Dimethylformamide; 15% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.31 (t, 1H, J = 6 Hz), 9.26 (d, 1H, J = 2 Hz),
8.78 (d, 1H, J = 2 Hz), 7.99 (d, 1H, J = 9 Hz), 7.46 (d, 1H, J = 2 Hz), 7.37 (t, 2H, J = 7 Hz), 7.35 (d, 2H, J = 7 Hz),
7.32 (dd, 1H, J = 9, 2 Hz), 7.25 (t, 1H, J = 7 Hz), 4.55 (d, 2H, J = 6 Hz), 3.94 (s, 3H); LC-MS (LC-ES) M+H = 293.
N-((1H-Pyrazol-5-yl)methyl)-7-methoxyquinoline-3-carboxamide 1v.

O
H
N
N
N
H
O
N
1v
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 1H-Pyrazole-3-methaneamine 8q (Oakwood); N,N-
Dimethylformamide; 43% yield; 1H NMR (400 MHz, CD3SOCD3) δ 12.65 (br s, 1H), 9.23 (d, 1H, J = 2 Hz), 9.18 (t,
1H, J = 6 Hz), 8.76 (d, 1H, J = 2 Hz), 7.99 (d, 1H, J = 9 Hz), 7.59 (br s, 1H), 7.45 (d, 1H, J = 2 Hz), 7.32 (dd, 1H, J
= 9, 2 Hz), 6.22 (t, 1H, J = 7 Hz), 4.53 (d, 2H, J = 6 Hz), 3.94 (s, 3H); LC-MS (LC-ES) M+H = 283.
7-Methoxy-N-((1-methyl-1H-pyrazol-5-yl)methyl)quinoline-3-carboxamide 1w.
O
N
N
N
H
O
N
1w
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); (1-Methyl-1H-pyrazol-5-yl)methylamine 8r
(Oakwood); N,N-Dimethylformamide; 32% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.23 (s, 1H), 9.22 (s, 1H), 8.76
(s, 1H), 7.98 (d, 1H, J = 9 Hz), 7.45 (s, 1H), 7.32 (s, 2H), 6.23 (s, 1H), 4.58 (d, 2H, J = 6 Hz), 3.94 (s, 3H), 3.84 (s,
3H); LC-MS (LC-ES) M+H = 297.
7-Methoxy-N-((1-methyl-1H-pyrazol-3-yl)methyl)quinoline-3-carboxamide 1x.
O
N
N
N
H
O
N
1x
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); [(1-Methyl-1H-pyrazol-3-yl)methyl]amine 8s
(Oakwood); N,N-Dimethylformamide; 39% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.23 (d, 1H, J = 2 Hz), 9.16 (t,
1H, J = 6 Hz), 8.76 (d, 1H, J = 2 Hz), 7.97 (d, 1H, J = 9 Hz), 7.59 (d, 1H, J = 2 Hz), 7.44 (d, 1H, J = 2 Hz), 7.32 (dd,
1H, J = 9, 2 Hz), 6.18 (d, 1H, J = 2 Hz), 4.47 (d, 2H, J = 6 Hz), 3.94 (s, 3H), 3.79 (s, 3H); LC-MS (LC-ES) M+H =
297.
7-Methoxy-N-((1-methyl-1H-imidazol-4-yl)methyl)quinoline-3-carboxamide 1y.
O
N
N
H
N
O
N
1y
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); (1-Methyl-1H-imidazol-4-yl)methylamine 8t
(Astatech); N,N-Dimethylformamide; 7% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.22 (d, 1H, J = 2 Hz), 9.34 (t,
1H, J = 6 Hz), 8.75 (d, 1H, J = 2 Hz), 7.97 (d, 1H, J = 9 Hz), 7.50 (d, 1H, J = 2 Hz), 7.44 (d, 1H, J = 2 Hz), 7.32 (dd,
1H, J = 9, 2 Hz), 7.01 (d, 1H, J = 2 Hz), 4.38 (d, 2H, J = 6 Hz), 3.94 (s, 3H), 3.60 (s, 3H); LC-MS (LC-ES) M+H =
297.
N-((4H-1,2,4-Triazol-3-yl)methyl)-7-methoxyquinoline-3-carboxamide 1z.

O
H
N
N
H
N
N
O
N
1z
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); (1H-1,2,4-Triazol-5-ylmethyl)amine 8u
1
(ChemBridge); N,N-Dimethylformamide; 2% yield; H NMR (400 MHz, CD3SOCD3) δ 9.50 (br s, 1H), 9.32 (br s,
1H), 9.24 (d, 1H, J = 2 Hz), 8.78 (d, 1H, J = 2 Hz), 8.19 (s, 1H), 7.99 (d, 1H, J = 9 Hz), 7.46 (d, 1H, J = 2 Hz), 7.33
(dd, 1H, J = 9, 2 Hz), 4.62 (s, 2H), 3.94 (s, 3H); LC-MS (LC-ES) M+H = 284.
N-((1H-Tetrazol-5-yl)methyl)-7-methoxyquinoline-3-carboxamide 1aa.
O
H
N
N
N
H
N
N
O
N
1aa
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); (1H-Tetrazol-5-ylmethyl)amine 8v (ChemBridge);
N,N-Dimethylformamide; 21% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.48 (t, 1H, J = 5 Hz), 9.24 (d, 1H, J = 2
Hz), 8.78 (d, 1H, J = 2 Hz), 7.99 (d, 1H, J = 9 Hz), 7.46 (d, 1H, J = 2 Hz), 7.34 (dd, 1H, J = 9, 2 Hz), 4.82 (d, 2H, J =
6 Hz), 3.94 (s, 3H); LC-MS (LC-ES) M+H = 285.
7-Methoxy-N-(pyrazin-2-ylmethyl)quinoline-3-carboxamide 1ab.
O
N
N
H
N
O
N
1ab
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 2-Aminomethylpyrazine 8w (Oakwood); N,N-
Dimethylformamide; 44% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.46 (t, 1H, J = 6 Hz), 9.25 (d, 1H, J = 2 Hz),
8.79 (d, 1H, J = 2 Hz), 8.71 (d, 1H, J = 2 Hz), 8.61 (dd, 1H, J = 2, 2 Hz), 8.55 (d, 1H, J = 2 Hz), 7.99 (d, 1H, J = 9
Hz), 7.46 (d, 1H, J = 2 Hz), 7.33 (dd, 1H, J = 9, 2 Hz), 4.69 (d, 2H, J = 6 Hz), 3.94 (s, 3H); LC-MS (LC-ES) M+H =
295.
7-Methoxy-N-(pyrimidin-4-ylmethyl)quinoline-3-carboxamide 1ac.
O
N
H
N
N
O
N
1ac
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 4-(Aminomethyl)pyrimidine 8x (Oakwood); N,N-
Dimethylformamide; 21% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.48 (t, 1H, J = 6 Hz), 9.28 (d, 1H, J = 2 Hz),
9.12 (s, 1H), 8.82 (d, 1H, J = 2 Hz), 8.74 (d, 1H, J = 5 Hz), 8.01 (d, 1H, J = 9 Hz), 7.52 (d, 1H, J = 5 Hz), 7.47 (d, 1H,
J = 2 Hz), 7.34 (dd, 1H, J = 9, 2 Hz), 4.62 (d, 2H, J = 6 Hz), 3.94 (s, 3H); LC-MS (LC-ES) M+H = 295.
7-Methoxy-N-(pyrimidin-5-ylmethyl)quinoline-3-carboxamide 1ad.
O
N
N
H
O
N
N
1ad

Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 5-Pyrimidinemethanamine 8y (Astatech); N,N-
Dimethylformamide; 1% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.37 (t, 1H, J = 6 Hz), 9.22 (d, 1H, J = 2 Hz), 9.09
(s, 1H), 8.83 (s, 1H), 8.76 (d, 1H, J = 2 Hz), 8.20 (s, 1H), 7.99 (d, 1H, J = 9 Hz), 7.45 (d, 1H, J = 2 Hz), 7.33 (dd, 1H,
J = 9, 2 Hz), 4.57 (d, 2H, J = 6 Hz), 3.94 (s, 3H); LC-MS (LC-ES) M+H = 295.
Racemic 7-Methoxy-N-(1-(pyrazin-2-yl)ethyl)quinoline-3-carboxamide 1ae.
O
N
N
H
N
O
N
1ae
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 1-Pyrazin-2-yl-ethylamine 8z (Oakwood); N,N-
Dimethylformamide; 49% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.26-9.20 (m, 2H), 8.81 (d, 1H, J = 2 Hz), 8.76
(br s, 1H), 8.61 (dd, 1H, J = 2, 2 Hz), 8.54 (d, 1H, J = 2 Hz), 7.99 (d, 1H, J = 9 Hz), 7.46 (d, 1H, J = 2 Hz), 7.32 (dd,
1H, J = 9, 2 Hz), 4.14 (q, 1H, J = 7 Hz), 3.95 (s, 3H), 1.59 (d, 3H, J = 7 Hz); LC-MS (LC-ES) M+H = 309.
Racemic 7-Methoxy-N-(1-(pyrimidin-4-yl)ethyl)quinoline-3-carboxamide 1af.
O
N
H
N
N
O
N
1af
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); [1-(4-Pyrimidinyl)ethyl]amine 8aa (ChemBridge);
N,N-Dimethylformamide; 45% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.26 (d, 1H, J = 1 Hz), 9.22 (d, 1H, J = 7
Hz), 9.14 (s, 1H), 8.82 (d, 1H, J = 2 Hz), 8.75 (d, 1H, J = 5 Hz), 8.01 (d, 1H, J = 9 Hz), 7.58 (dd, 1H, J = 5, 1 Hz),
7.47 (d, 1H, J = 2 Hz), 7.34 (dd, 1H, J = 9, 2 Hz), 4.14 (q, 1H, J = 7 Hz), 3.95 (s, 3H), 1.56 (d, 3H, J = 7 Hz); LC-MS
(LC-ES) M+H = 309.
7-Methoxy-N-((2-oxo-1,2-dihydropyridin-4-yl)methyl)quinoline-3-carboxamide 1ag.
O
O
N
H
NH
O
N
1ag
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 4-(Aminomethyl)-2(1H)-pyridinone 8ab
(Astatech); N,N-Dimethylformamide; 27% yield; 1H NMR (400 MHz, CD3SOCD3) δ 11.46 (br s, 1H), 9.29 (t, 1H, J
= 6 Hz), 9.24 (d, 1H, J = 2 Hz), 8.79 (d, 1H, J = 2 Hz), 8.01 (d, 1H, J = 9 Hz), 7.46 (d, 1H, J = 2 Hz), 7.33 (dd, 1H, J
= 9, 2 Hz), 7.32 (d, 1H, J = 7 Hz), 6.23 (s, 1H), 6.17 (dd, 1H, J = 7, 1 Hz), 4.35 (d, 2H, J = 6 Hz), 3.95 (s, 3H); LC-
MS (LC-ES) M+H = 310.
N-Cyclopentyl-7-methoxyquinoline-3-carboxamide 1ah.
O
N
H
O
N
1ah
Commercial (Aurora Building Blocks); Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech);
Cyclopentamine 8ac (Aldrich); N,N-Dimethylformamide; 67% yield; 1H NMR (400 MHz, CD3OD) δ 9.18 (d, 1H, J
= 2 Hz), 8.81 (d, 1H, J = 2 Hz), 8.06 (br s, 1H), 7.99 (d, 1H, J = 9 Hz), 7.42 (d, 1H, J = 2 Hz), 7.38 (dd, 1H, J = 9, 2

Hz), 4.42-4.32 (m, 1H), 4.00 (s, 3H), 2.14-2.02 (m, 2H), 1.86-1.76 (m, 2H), 1.72-1.56 (m, 4H); LC-MS (LC-ES) M+H
= 271.
N-Cyclohexyl-7-methoxyquinoline-3-carboxamide 1ai.
O
N
H
O
N
1ai
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); Cyclohexylamine 8ad (Aldrich); N,N-
Dimethylformamide; 49% yield; 1H NMR (400 MHz, CD3OD) δ 9.19 (d, 1H, J = 2 Hz), 8.83 (d, 1H, J = 2 Hz), 8.06
(br s, 1H), 8.00 (d, 1H, J = 9 Hz), 7.43 (d, 1H, J = 2 Hz), 7.39 (dd, 1H, J = 9, 2 Hz), 4.01 (s, 3H), 3.98-3.86 (m, 1H),
2.06-1.96 (m, 2H), 1.88-1.76 (m, 2H), 1.74-1.64 (m, 1H), 1.58-1.34 (m, 4H), 1.32-1.18 (m, 1H); LC-MS (LC-ES)
M+H = 285.
7-Methoxy-N-(tetrahydro-2H-pyran-4-yl)quinoline-3-carboxamide 1aj.
O
O
N
H
O
N
1aj
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 4-Aminotetrahydropyran 8ae (Aldrich); N,N-
Dimethylformamide; 39% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.20 (d, 1H, J = 2 Hz), 8.72 (d, 1H, J = 2 Hz),
8.59 (br s, 1H), 7.99 (d, 1H, J = 9 Hz), 7.44 (d, 1H, J = 2 Hz), 7.32 (dd, 1H, J = 9, 2 Hz), 4.12-4.00 (m, 1H), 3.93 (s,
3H), 3.89 (br d, 2H, J = 11 Hz), 3.40 (br t, 2H, J = 13 Hz), 1.80 (br d, 2H, J = 13 Hz), 1.66-1.56 (m, 2H); LC-MS (LC-
ES) M+H = 287.
N-(trans-4-Hydroxycyclohexyl)-7-methoxyquinoline-3-carboxamide 1ak.
OH
O
N
H
O
N
1ak
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); trans-4-Aminocyclohexanol 8af (Aldrich);
Dichloromethane:N,N-Dimethylformamide (2:1); 79% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.18 (d, 1H, J = 2
Hz), 8.72 (d, 1H, J = 2 Hz), 8.45 (d, 1H, J = 8 Hz), 7.98 (d, 1H, J = 9 Hz), 7.45 (d, 1H, J = 2 Hz), 7.32 (dd, 1H, J = 9,
2 Hz), 4.58 (d, 1H, J = 6 Hz), 3.94 (s, 3H), 3.82-3.72 (m, 1H), 3.46-3.36 (m, 1H), 1.92-1.82 (m, 4H), 1.44-1.32 (m,
2H), 1.32-1.20 (m, 2H); LC-MS (LC-ES) M+H = 301.
N-(1-Acetylpiperidin-4-yl)-7-methoxyquinoline-3-carboxamide 1al.
O
O
N
N
H
O
N
1al
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 1-Acetyl-4-aminopiperidine 8ag (Astatech); N,N-
Dimethylformamide; 78% yield; 1H NMR (400 MHz, CD3OD) δ 9.14 (d, 1H, J = 2 Hz), 8.67 (d, 1H, J = 2 Hz), 8.09
(br s, 1H), 7.89 (d, 1H, J = 9 Hz), 7.39 (d, 1H, J = 2 Hz), 7.30 (dd, 1H, J = 9, 2 Hz), 4.58-4.48 (m, 1H), 4.24-4.12 (m,

1H), 4.02-3.92 (m, 1H), 3.95 (s, 3H), 3.32-3.22 (m, 1H), 2.88-2.76 (m, 1H), 2.14-1.96 (m, 2H), 2.11 (s, 3H), 1.66-
1.46 (m, 2H); LC-MS (LC-ES) M+H = 328.
7-Methoxy-N-(1-(methylsulfonyl)piperidin-4-yl)quinoline-3-carboxamide 1am.
O
O
S
O
N
N
H
O
N
1am
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 4-Amino-1-methanesulfonylpiperidine 8ah
1
(Oakwood); Dichloromethane:N,N-Dimethylformamide (2:1); 75% yield; H NMR (400 MHz, CD3OD) δ 9.21 (d,
1H, J = 2 Hz), 8.73 (d, 1H, J = 2 Hz), 8.63 (br d, 1H, J = 7 Hz), 8.00 (d, 1H, J = 9 Hz), 7.46 (d, 1H, J = 2 Hz), 7.33
(dd, 1H, J = 9, 2 Hz), 4.06-3.92 (m, 1H), 3.95 (s, 3H), 3.64-3.54 (m, 2H), 2.94-2.84 (m, 2H), 2.88 (s, 3H), 2.02-1.92
(m, 2H), 1.70-1.54 (m, 2H); LC-MS (LC-ES) M+H = 364.
7-Methoxy-N-(2-oxopiperidin-3-yl)quinoline-3-carboxamide 1an.
H
O
N
O
N
H
O
N
1an
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 3-Aminopiperidin-2-one 8ai (Aldrich); N,N-
Dimethylformamide; 56% yield; 1H NMR (400 MHz, CD3OD) δ 9.22 (d, 1H, J = 2 Hz), 8.91 (d, 1H, J = 8 Hz), 8.75
(br d, 1H, J = 2 Hz), 7.99 (d, 1H, J = 9 Hz), 7.69 (br s, 1H), 7.46 (d, 1H, J = 2 Hz), 7.33 (dd, 1H, J = 9, 2 Hz), 4.48-
4.42 (m, 1H), 3.95 (s, 3H), 3.22-3.14 (m, 2H), 2.10-2.02 (m, 1H), 1.90-1.76 (m, 3H); LC-MS (LC-ES) M+H = 300.
7-Methoxy-N-(2-(2-oxoimidazolidin-1-yl)ethyl)quinoline-3-carboxamide 1ao.
O
NH
N
N
H
O
O
N
1ao
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 1-(2-Aminoethyl)-2-imidazolidinone 8aj
(Oakwood); N,N-Dimethylformamide; 34% yield; 1H NMR (400 MHz, CD3OD) δ 9.18 (d, 1H, J = 2 Hz), 8.81 (t, 1H,
J = 6 Hz), 8.69 (d, 1H, J = 2 Hz), 7.98 (d, 1H, J = 9 Hz), 7.45 (d, 1H, J = 2 Hz), 7.32 (dd, 1H, J = 9, 2 Hz), 6.29 (br s,
1H), 3.95 (s, 3H), 3.46-3.88 (m, 4H), 3.26 (t, 2H, J = 7 Hz), 3.22 (t, 2H, J = 7 Hz); LC-MS (LC-ES) M+H = 315.
N-(2-Amino-2-oxoethyl)-7-methoxyquinoline-3-carboxamide 1ap.
O
NH2
N
H
O
O
N
1ap
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 2-Aminoacetamide 8ak (Aldrich); N,N-
Dimethylformamide; 52% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.24 (d, 1H, J = 2 Hz), 8.92 (t, 1H, J = 6 Hz),
8.76 (d, 1H, J = 2 Hz), 7.99 (d, 1H, J = 9 Hz), 7.46 (d, 1H, J = 2 Hz), 7.42 (br s, 1H), 7.33 (dd, 1H, J = 9, 2 Hz), 7.06
(br s, 1H), 3.95 (s, 3H), 3.88 (d, 2H, J = 6 Hz); LC-MS (LC-ES) M+H = 260.

(S)-N-(1-Amino-1-oxopropan-2-yl)-7-methoxyquinoline-3-carboxamide 1aq.
O
NH2
N
H
O
O
N
1aq
Method A; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); (S)-2-Aminopropanamide 8al (Aldrich);
dichloromethane; 56% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.23 (d, 1H, J = 2 Hz), 8.79 (d, 1H, J = 2 Hz), 8.73
(d, 1H, J = 8 Hz), 7.98 (d, 1H, J = 9 Hz), 7.45 (d, 1H, J = 2 Hz), 7.45 (br s, 1H), 7.32 (dd, 1H, J = 9, 2 Hz), 7.03 (br
s, 1H), 4.46 (p, 1H, J = 7 Hz), 3.94 (s, 3H), 1.36 (d, 3H, J = 7 Hz); LC-MS (LC-ES) M+H = 274.
(R)-N-(1-Amino-1-oxopropan-2-yl)-7-methoxyquinoline-3-carboxamide 1ar.
O
NH2
N
H
O
O
N
1ar
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); (R)-2-Aminopropanamide 8am (Aldrich); N,N-
Dimethylformamide; 47% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.23 (d, 1H, J = 2 Hz), 8.79 (d, 1H, J = 2 Hz),
8.67 (d, 1H, J = 8 Hz), 7.98 (d, 1H, J = 9 Hz), 7.45 (d, 1H, J = 2 Hz), 7.41 (br s, 1H), 7.32 (dd, 1H, J = 9, 2 Hz),
6.98 (br s, 1H), 4.46 (p, 1H, J = 7 Hz), 3.94 (s, 3H), 1.36 (d, 3H, J = 7 Hz); LC-MS (LC-ES) M+H = 274.
(S)-7-Methoxy-N-(tetrahydrofuran-3-yl)quinoline-3-carboxamide 1as.
O
O
N
H
O
N
1as
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); (S)-tetrahydrofuran-3-amine 8an (Aldrich); N,N-
Dimethylformamide; 72% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.21 (d, 1H, J = 2 Hz), 8.77 (br d, 1H, J = 7
Hz), 8.74 (d, 1H, J = 2 Hz), 7.98 (d, 1H, J = 9 Hz), 7.45 (d, 1H, J = 2 Hz), 7.33 (dd, 1H, J = 9, 2 Hz), 4.58-4.46 (m,
1H), 3.95 (s, 3H), 3.92-3.84 (m, 2H), 3.74 (dt, 1H, J = 9, 7 Hz), 3.64 (dd, 1H, J = 9, 4 Hz), 2.26-2.14 (m, 1H), 2.02-
1.90 (m, 1H); LC-MS (LC-ES) M+H = 273.
(S)-7-Methoxy-N-(2-oxopyrrolidin-3-yl)quinoline-3-carboxamide 1at.
O
NH
O
N
H
O
N
1at
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 3-Amino-2-pyrrolidinone 8ao (Astatech); N,N-
Dimethylformamide; 53% yield; Separation of enantiomers on a Chiralpak® IB column (1:3 ethanol:heptane, 1.0
mL/min, 25 minutes, tR = 11.7 minutes); 1H NMR (400 MHz, CD3OD) δ 9.22 (d, 1H, J = 2 Hz), 8.76 (d, 1H, J = 2
Hz), 7.96 (d, 1H, J = 9 Hz), 7.46 (d, 1H, J = 2 Hz), 7.36 (dd, 1H, J = 9, 2 Hz), 4.02 (s, 3H), 4.80-4.72 (m, 1H), 3.48-
3.32 (m, 2H), 2.64-2.52 (m, 1H), 2.28-2.12 (m, 1H); LC-MS (LC-ES) M+H = 286.
(R)-7-Methoxy-N-(2-oxopyrrolidin-3-yl)quinoline-3-carboxamide 1au.

O
NH
O
N
H
O
N
1au
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 3-Amino-2-pyrrolidinone 8ap (Astatech); N,N-
Dimethylformamide; 53% yield; Separation of enantiomers on a Chiralpak® IB column (1:3 ethanol:heptane, 1.0
mL/min, 25 minutes, tR = 16.6 minutes); 1H NMR (400 MHz, CD3OD) δ 9.22 (d, 1H, J = 2 Hz), 8.76 (d, 1H, J = 2
Hz), 7.96 (d, 1H, J = 9 Hz), 7.46 (d, 1H, J = 2 Hz), 7.36 (dd, 1H, J = 9, 2 Hz), 4.02 (s, 3H), 4.80-4.72 (m, 1H), 3.48-
3.32 (m, 2H), 2.64-2.52 (m, 1H), 2.28-2.12 (m, 1H); LC-MS (LC-ES) M+H = 286.
(S)-7-Methoxy-N-(1-methyl-2-oxopyrrolidin-3-yl)quinoline-3-carboxamide 1av.
O
N
O
N
H
O
N
1av
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); (S)-3-amino-1-methylpyrrolidin-2-one 8aq
(PharmaBlock); N,N-Dimethylformamide; 17% yield; 1H NMR (400 MHz, CD3OD) δ 9.17 (d, 1H, J = 2 Hz), 8.70
(d, 1H, J = 2 Hz), 7.91 (d, 1H, J = 9 Hz), 7.41 (d, 1H, J = 2 Hz), 7.31 (dd, 1H, J = 9, 2 Hz), 4.74 (t, 1H, J = 7 Hz),
3.98 (s, 3H), 3.52-3.44 (m, 2H), 2.91 (s, 3H), 2.60-2.48 (m, 1H), 2.18-2.04 (m, 1H); LC-MS (LC-ES) M+H = 300.
(S)-7-Methoxy-N-(3-oxoisoxazolidin-4-yl)quinoline-3-carboxamide 1aw.
O
NH
O
O
N
H
O
N
1aw
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); L-Cycloserine 8ar (Aldrich); N,N-
Dimethylformamide; 28% yield; 1H NMR (400 MHz, CD3SOCD3) δ 11.62 (br s, 1H), 9.22 (d, 1H, J = 2 Hz), 8.77
(d, 1H, J = 2 Hz), 8.13 (br s, 1H), 8.00 (d, 1H, J = 9 Hz), 7.46 (d, 1H, J = 2 Hz), 7.34 (dd, 1H, J = 9, 2 Hz), 5.40 (q,
1H, J = 7 Hz), 4.62 (t, 1H, J = 7 Hz), 4.16 (t, 1H, J = 7 Hz), 3.95 (s, 3H); LC-MS (LC-ES) M+H = 288.
N-((3S,4R)-4-Hydroxy-2-oxopyrrolidin-3-yl)-7-methoxyquinoline-3-carboxamide 1ax.
O
NH
O
N
H
OH
O
N
1ax
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); (3S,4R)-3-Amino-4-hydroxypyrrolidin-2-one
8as;40 N,N-Dimethylformamide; 51% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.24 (d, 1H, J = 2 Hz), 8.95 (br d,
1H, J = 8 Hz), 8.76 (d, 1H, J = 2 Hz), 8.01 (d, 1H, J = 9 Hz), 7.81 (br s, 1H), 7.47 (d, 1H, J = 2 Hz), 7.34 (dd, 1H, J
= 9, 2 Hz), 5.56 (d, 1H, J = 5 Hz), 4.46-4.34 (m, 2H), 3.96 (s, 3H), 3.50-3.44 (m, 1H), 3.04-2.96 (m, 1H); LC-MS
(LC-ES) M+H = 302.
Racemic 7-Methoxy-N-(3-methyl-2-oxopyrrolidin-3-yl)quinoline-3-carboxamide 1ay.

O
NH
O
N
H
O
N
1ay
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 3-Amino-3-methylpyrrolidin-2-one 8at;41
Dichloromethane:N,N-Dimethylformamide (1:1); 18% yield; 1H NMR (400 MHz, CD3OD) δ 9.16 (d, 1H, J = 2 Hz),
8.73 (d, 1H, J = 2 Hz), 8.06 (br s, 1H), 7.92 (d, 1H, J = 9 Hz), 7.41 (d, 1H, J = 2 Hz), 7.32 (dd, 1H, J = 9, 2 Hz),
3.98 (s, 3H), 3.50 (dt, 1H, J = 9, 2 Hz), 3.40 (dt, 1H, J = 9, 7 Hz), 2.79 (dt, 1H, J = 9, 7 Hz), 2.17 (ddd, 1H, J = 9, 7,
2 Hz), 1.52 (s, 3H); LC-MS (LC-ES) M+H = 300.
7-Methoxy-N-(2-oxoindolin-3-yl)quinoline-3-carboxamide 1az.
O
NH
O
N
H
O
N
1az
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 3-Aminoindolin-2-one 8au (Astatech); N,N-
Dimethylformamide; 63% yield; 1H NMR (400 MHz, CD3OD) δ 9.21 (d, 1H, J = 2 Hz), 8.75 (br s, 1H), 7.94 (d, 1H,
J = 9 Hz), 7.81 (br s, 1H), 7.43 (d, 1H, J = 2 Hz), 7.34 (dd, 1H, J = 9, 2 Hz), 7.32-7.22 (m, 2H), 7.03 (t, 1H, J = 7
Hz), 6.93 (d, 1H, J = 7 Hz), 5.46 (s, 1H), 3.98 (s, 3H); LC-MS (LC-ES) M+H = 334.
N-(2,3-Dihydro-1H-inden-1-yl)-7-(methyloxy)-3-quinolinecarboxamide 1ba.
O
N
H
O
N
1ba
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 1-Aminoindane 8av (Aldrich); N,N-
Dimethylformamide; 82% yield; 1H NMR (400 MHz, CDCl3) δ 9.20 (d, 1H, J = 2 Hz), 8.67 (d, 1H, J = 2 Hz), 8.16
(br s, 1H), 7.81 (d, 1H, J = 9 Hz), 7.49 (d, 1H, J = 2 Hz), 7.39 (br d, 1H, J = 7 Hz), 7.29 (dd, 1H, J = 9, 2 Hz), 7.28-
7.20 (m, 2H), 6.82 (br d, 1H, J = 7 Hz), 5.75 (q, 1H, J = 7 Hz), 3.97 (s, 3H), 3.14-3.02 (m, 1H), 3.00-2.90 (m, 1H),
2.78-2.66 (m, 1H), 2.08-1.96 (m, 1H); LC-MS (LC-ES) M+H = 319.
(S)-7-Methoxy-N-(5-oxopyrrolidin-3-yl)quinoline-3-carboxamide 1bb.
NH
O
O
N
H
O
N
1bb
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 4-Aminopyrrolidin-2-one 8aw (Fluorochem);
N,N-Dimethylformamide; Separation of enantiomers on a Chiralcel® OJ column (1:4 ethanol:heptane, 1.0 mL/min,
30 minutes, tR = 10.6 minutes); 56% yield; 1H NMR (400 MHz, CD3OD) δ 9.16 (d, 1H, J = 2 Hz), 8.73 (d, 1H, J = 2
Hz), 8.06 (br s, 1H), 7.92 (d, 1H, J = 9 Hz), 7.39 (d, 1H, J = 2 Hz), 7.32 (dd, 1H, J = 9, 2 Hz), 4.86-4.74 (m, 1H),
3.97 (s, 3H), 3.83 (dd, 1H, J = 9, 7 Hz), 3.41 (dd, 1H, J = 9, 4 Hz), 2.79 (dd, 1H, J = 13, 7 Hz), 2.47 (dd, 1H, J = 13,
4 Hz); LC-MS (LC-ES) M+H = 286.
(R)-7-Methoxy-N-(5-oxopyrrolidin-3-yl)quinoline-3-carboxamide 1bc.

NH
O
O
N
H
O
N
1bc
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 4-Aminopyrrolidin-2-one 8ax (Fluorochem); N,N-
Dimethylformamide; Separation of enantiomers on a Chiralcel® OJ column (1:4 ethanol:heptane, 1.0 mL/min, 30
minutes, tR = 15.9 minutes); 56% yield; 1H NMR (400 MHz, CD3OD) δ 9.16 (d, 1H, J = 2 Hz), 8.73 (d, 1H, J = 2
Hz), 8.06 (br s, 1H), 7.92 (d, 1H, J = 9 Hz), 7.39 (d, 1H, J = 2 Hz), 7.32 (dd, 1H, J = 9, 2 Hz), 4.86-4.74 (m, 1H),
3.97 (s, 3H), 3.83 (dd, 1H, J = 9, 7 Hz), 3.41 (dd, 1H, J = 9, 4 Hz), 2.79 (dd, 1H, J = 13, 7 Hz), 2.47 (dd, 1H, J = 13,
4 Hz); LC-MS (LC-ES) M+H = 286.
7-Methoxy-N-(piperidin-4-yl)quinoline-3-carboxamide 1bd.
O
NH
N
H
O
N
1bd
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 4-Amino-1-N-Boc-piperidine 8ay (Astatech);
N,N-Dimethylformamide; Followed by hydrochloric acid in 1,4-dioxane cleavage of the tert-butyloxycarbonyl
protecting group; 82% yield; 1H NMR (400 MHz, CD3OD) δ 9.57 (br s, 1H), 9.50 (br s, 1H), 8.34 (d, 1H, J = 2 Hz),
8,01 (br s, 2H), 7.66 (d, 1H, J = 2 Hz), 7.58 (br s, 1H), 4.34-4.22 (m, 1H), 4.12 (s, 3H), 3.58-3.48 (d, 2H, J = 12 Hz),
3.18 (t, 2H, J = 12 Hz), 2.26 (d, 2H, J = 13 Hz), 2.12-1.96 (m, 2H); LC-MS (LC-ES) M+H = 286.
7-Methoxy-N-(1-(1-methyl-1H-tetrazol-5-yl)piperidin-4-yl)quinoline-3-carboxamide 1be.
N
N
N
N
O
N
N
H
O
N
1be
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 1-(1-Methyl-1H-tetrazol-5-yl)piperidin-4-amine
8az (Enamine); N,N-Dimethylformamide; 82% yield; 1H NMR (400 MHz, CDCl3) δ 9.22 (d, 1H, J = 2 Hz), 8.57 (d,
1H, J = 2 Hz), 7.81 (d, 1H, J = 9 Hz), 7.42 (d, 1H, J = 2 Hz), 7.25 (dd, 1H, J = 9, 2 Hz), 6.74 (d, 1H, J = 8 Hz),
4.42-4.19 (m, 1H), 3.97 (s, 3H), 3.90 (s, 3H), 3.68 (d, 2H, J = 13 Hz), 3.37-3.17 (m, 2H), 2.26-2.10 (m, 2H), 1.95-
1.76 (m, 2H); LC-MS (LC-ES) M+H = 368.
7-Methoxy-N-(1-(thiazol-2-yl)piperidin-4-yl)quinoline-3-carboxamide 1bf.
N
S
O
N
N
H
O
N
1bf
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 1-(1,3-thiazol-2-yl)piperidin-4-amine 8ba
(PharmaBlock); Dichloromethane:N,N-Dimethylformamide (3:1); 34% yield; 1H NMR (400 MHz, CDCl3) δ 9.21 (d,
1H, J = 2 Hz), 8.73 (d, 1H, J = 2 Hz), 8.56 (d, 1H, J = 7 Hz), 7.98 (d, 1H, J = 9 Hz), 7.45 (d, 1H, J = 2 Hz), 7.32 (dd,
1H, J = 9, 2 Hz), 7.16 (d, 1H, J = 4 Hz), 6.83 (d, 1H, J = 4 Hz), 4.20-4.06 (m, 1H), 3.95 (s, 3H), 3.94 (d, 2H, J = 13
Hz), 3.18 (br t, 2H, J = 12 Hz), 1.95 (br d, 2H, J = 12 Hz), 1.68 (br q, 2H, J = 12 Hz); LC-MS (LC-ES) M+H = 369.

N-(trans-4-(2-Hydroxypropan-2-yl)cyclohexyl)-7-methoxyquinoline-3-carboxamide 1bg.
O
OH
N
H
O
N
1bg
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 2-(trans-4-Aminocyclohexyl)propan-2-ol 8bb
1
(PharmaBlock); Dichloromethane:N,N-Dimethylformamide (1.6:1); 71% yield; H NMR (400 MHz, CD3SOCD3) δ
9.27 (d, 1H, J = 2 Hz), 8.91 (d, 1H, J = 2 Hz), 8.56 (d, 1H, J = 8 Hz), 8.08 (d, 1H, J = 9 Hz), 7.48 (d, 1H, J = 2 Hz),
7.40 (dd, 1H, J = 9, 2 Hz), 3.97 (s, 3H), 3.77 (dtt, 1H, J = 12, 8, 4 Hz), 1.96 (br d, 2H, J = 13 Hz), 1.85 (br d, 2H, J =
11 Hz), 1.33 (q, 2H, J =12 Hz), 1.26-1.06 (m, 3H), 1.05 (s, 6H); LC-MS (LC-ES) M+H = 343.
7-Methoxy-N-(trans-4-(methylcarbamoyl)cyclohexyl)quinoline-3-carboxamide 1bh.
O
O
N
H
N
H
O
N
1bh
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); trans-4-Amino-N-methylcyclohexanecarboxamide
8bc (BOC Sciences); Dichloromethane; 31% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.11 (d, 1H, J = 2 Hz), 8.70
(d, 1H, J = 2 Hz), 8.44 (d, 1H, J = 8 Hz), 7.98 (d, 1H, J = 9 Hz), 7.63 (br q, 1H, J = 5 Hz), 7.45 (d, 1H, J = 2 Hz), 7.31
(dd, 1H, J = 9, 2 Hz), 3.95 (s, 3H), 3.86-3.72 (m, 1H), 2.57 (d, 3H, J = 5 Hz), 2.08 (tt, 1H, J = 15, 3 Hz), 1.94 (br d,
2H, J = 13 Hz), 1.79 (br d, 2H, J = 12 Hz), 1.47 (br q, 2H, J = 13 Hz), 1.37 (br q, 2H, J = 13 Hz); LC-MS (LC-ES)
M+H = 342.
N-(trans-4-Hydroxy-4-methylcyclohexyl)-7-methoxyquinoline-3-carboxamide 1bi.
OH
O
N
H
O
N
1bi
Method A; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); trans-4-Amino-1-methylcyclohexanol 8bd
(Enamine); 1,4-Dioxane; 49% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.17 (d, 1H, J = 2 Hz), 8.69 (d, 1H, J = 2
Hz), 8.41 (d, 1H, J = 8 Hz), 7.98 (d, 1H, J = 9 Hz), 7.44 (d, 1H, J = 2 Hz), 7.31 (dd, 1H, J = 9, 2 Hz), 4.29 (s, 1H),
3.94 (s, 3H), 3.92-3.80 (m, 1H), 1.86-1.76 (m, 2H), 1.66-1.56 (m, 2H), 1.56-1.40 (m, 4H), 1.16 (s, 3H); LC-MS (LC-
ES) M+H = 315.
N-(6-(2-Hydroxypropan-2-yl)spiro[3.3]heptan-2-yl)-7-methoxyquinoline-3-carboxamide 1bj.
OH
O
N
H
O
N
1bj
Method A; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 2-(6-Aminospiro[3.3]heptan-2-yl)propan-2-ol
1
8be;41 1,4-Dioxane; 74% yield; H NMR (400 MHz, CD3SOCD3) δ 9.18 (d, 1H, J = 2 Hz), 8.77 (d, 1H, J = 7 Hz),

8.70 (t, 1H, J = 2 Hz), 7.97 (d, 1H, J = 9 Hz), 7.44 (d, 1H, J = 2 Hz), 7.31 (dd, 1H, J = 9, 2 Hz), 4.33 (h, 1H, J = 8
Hz), 3.99 (s, 1H), 3.94 (s, 3H), 2.46-2.36 (m, 1H), 2.22-2.12 (m, 1H), 2.11 (q, 2H, J = 9 Hz), 2.04-1.86 (m, 4H), 1.76-
1.66 (m, 1H), 0.96 (s, 3H), 0.94 (s, 3H); LC-MS (LC-ES) M+H = 355.
N-(trans-4-(2-Hydroxy-2-methylpropoxy)cyclohexyl)-7-methoxyquinoline-3-carboxamide 1bk.
O
O
OH
N
H
O
N
1bk
Method A; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); 1-((trans-4-Aminocyclohexyl)oxy)-2-
methylpropan-2-ol 8bf;41 1,4-Dioxane; 50% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.18 (d, 1H, J = 2 Hz), 8.69
(d, 1H, J = 2 Hz), 8.45 (d, 1H, J = 8 Hz), 7.98 (d, 1H, J = 9 Hz), 7.44 (d, 1H, J = 2 Hz), 7.31 (dd, 1H, J = 9, 2 Hz),
4.21 (s, 1H), 3.94 (s, 3H), 3.81 (qt, 1H, J = 8, 4 Hz), 3.28-3.18 (m, 1H), 3.17 (s, 2H), 2.02 (br d, 2H, J = 10 Hz), 1.90
(br d, 2H, J = 10 Hz), 1.39 (q, 2H, J = 13 Hz), 1.26 (q, 2H, J = 13 Hz), 1.06 (s, 6H); LC-MS (LC-ES) M+H = 373.
7-Methoxy-N-(trans-4-((2,2,2-trifluoroethyl)amino)cyclohexyl)quinoline-3-carboxamide 1bl.
F
H
F
F
N
O
N
H
O
N
1bl
Method A; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); trans-N1-(2,2,2-Trifluoroethyl)cyclohexane-1,4-
diamine 8bg;41 1,4-Dioxane; 43% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.18 (d, 1H, J = 2 Hz), 8.69 (d, 1H, J =
2 Hz), 8.45 (d, 1H, J = 8 Hz), 7.97 (d, 1H, J = 9 Hz), 7.44 (d, 1H, J = 2 Hz), 7.31 (dd, 1H, J = 9, 2 Hz), 3.94 (s, 3H),
3.78 (qt, 1H, J = 8, 4 Hz), 3.30-3.18 (m, 2H), 2.48-2.36 (m, 1H), 2.20 (q, 1H, J = 8 Hz), 1.91 (br t, 4H, J = 14 Hz),
1.36 (q, 2H, J = 12 Hz), 1.11 (q, 2H, J = 14 Hz); LC-MS (LC-ES) M+H = 382.
N-((1s,3s)-3-Hydroxycyclobutyl)-7-methoxyquinoline-3-carboxamide 1bm.
OH
O
N
H
O
N
1bm
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); cis-3-Aminocyclobutanol 8bh (Astatech); N,N-
Dimethylformamide; 21% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.18 (d, 1H, J = 2 Hz), 8.79 (d, 1H, J = 7 Hz),
8.71 (d, 1H, J = 2 Hz), 7.97 (d, 1H, J = 9 Hz), 7.44 (d, 1H, J = 2 Hz), 7.32 (dd, 1H, J = 9, 2 Hz), 5.14 (br s, 1H), 4.02-
3.90 (m, 1H), 3.94 (s, 3H), 3.88 (p, 1H, J = 8 Hz), 2.64-2.54 (m, 2H), 2.00-1.88 (m, 2H); LC-MS (LC-ES) M+H =
273.
N-((1r,3r)-3-Hydroxycyclobutyl)-7-methoxyquinoline-3-carboxamide 1bn.
OH
O
N
H
O
N
1bn
Method B; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); trans-3-Aminocyclobutanol 8bi (Astatech); N,N-
Dimethylformamide; 36% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.19 (d, 1H, J = 2 Hz), 8.80 (d, 1H, J = 7 Hz),

8.71 (d, 1H, J = 2 Hz), 7.97 (d, 1H, J = 9 Hz), 7.44 (d, 1H, J = 2 Hz), 7.32 (dd, 1H, J = 9, 2 Hz), 5.14 (br s, 1H), 4.02-
3.90 (m, 1H), 3.94 (s, 3H), 3.88 (p, 1H, J = 8 Hz), 2.64-2.54 (m, 2H), 2.00-1.90 (m, 2H); LC-MS (LC-ES) M+H =
273.
N-((1s,3s)-3-Hydroxy-3-methylcyclobutyl)-7-methoxyquinoline-3-carboxamide 1bo.
O
OH
N
H
O
N
1bo
Method A; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); cis-3-Amino-1-methylcyclobutanol 8bj
(Astatech); 1,4-Dioxane; 26% yield; 1H NMR (400 MHz, CD3SOCD3) δ 9.19 (d, 1H, J = 2 Hz), 8.82 (d, 1H, J = 7
Hz), 8.72 (d, 1H, J = 2 Hz), 7.97 (d, 1H, J = 9 Hz), 7.44 (d, 1H, J = 2 Hz), 7.31 (dd, 1H, J = 9, 2 Hz), 4.98 (s, 1H),
4.02 (h, 1H, J = 8 Hz), 3.94 (s, 3H), 2.36-2.28 (m, 2H), 2.16-2.10 (m, 2H), 1.28 (s, 3H); LC-MS (LC-ES) M+H =
287.
N-((1r,3r)-3-Hydroxy-3-methylcyclobutyl)-7-methoxyquinoline-3-carboxamide 1bp.
OH
O
N
H
O
N
1bp
Method A; 7-Methoxyquinoline-3-carboxylic acid 7a (Astatech); trans-3-Amino-1-methylcyclobutanol 8bk
1
(Astatech); 1,4-Dioxane; 25% yield; H NMR (400 MHz, CD3SOCD3) δ 9.18 (d, 1H, J = 2 Hz), 8.79 (d, 1H, J = 7
Hz), 8.70 (d, 1H, J = 2 Hz), 7.98 (d, 1H, J = 9 Hz), 7.44 (d, 1H, J = 2 Hz), 7.31 (dd, 1H, J = 9, 2 Hz), 4.86 (s, 1H),
4.54 (h, 1H, J = 8 Hz), 3.94 (s, 3H), 2.34-2.26 (m, 2H), 2.12-2.06 (m, 2H), 1.29 (s, 3H); LC-MS (LC-ES) M+H = 287.
Diethyl 2-(((3-(trifluoromethoxy)phenyl)amino)methylene)malonate 4a
F
O
F
F
O
N
H
O
O
O
4a
Diethyl 2-(ethoxymethylene)malonate 3 (5.70 mL, 28.2 mmol) was added to 3-(trifluoromethoxy)aniline 2a (3.77 mL,
28.2 mmol) in toluene (30 mL) and the reaction mixture was heated in a sealed tube at 120 °C for eighteen hours, then
concentrated to give diethyl 2-(((3-(trifluoromethoxy)phenyl)amino)methylene)malonate 4a (10.0 g, 28.8 mmol, 99%
1
yield) as a dark red syrup. H NMR (400 MHz, CDCl3) δ 11.05 (d, 1H, J = 14 Hz), 8.47 (d, 1H, J = 13 Hz), 7.44-7.37
(m, 1H), 7.07 (ddd, 1H, J = 8, 2, 1 Hz), 7.04-6.96 (m, 2H), 4.32 (q, 2H, J = 7 Hz), 4.27 (q, 2H, J = 7 Hz), 1.39 (t, 3H,
J = 7 Hz), 1.34 (t, 3H, J = 7 Hz); LC-MS (LC-ES) M+H = 348.
Ethyl 4-chloro-7-(trifluoromethoxy)quinoline-3-carboxylate 5a
Cl
O
F
O
F
F
O
N
5a
Phosphorus
oxychloride
(30.2
ml,
324
mmol)
was
added
to
diethyl
2-(((3-
(trifluoromethoxy)phenyl)amino)methylene)malonate 4a (9.00 g, 25.9 mmol) and the reaction mixture was heated in
a sealed tube at 115 °C. After three hours, the temperature was increased to 125 °C and the reaction mixture was

heated for fifteen hours. Upon cooling, the volatiles were removed in vacuo. The residue was dissolved in
dichloromethane, washed with saturated sodium bicarbonate and brine, dried over sodium sulfate and concentrated.
The residue was purified by flash chromatography, eluting with ethyl acetate:hexanes (1:49 to 1:1) to afford ethyl 4-
1
chloro-7-(trifluoromethoxy)quinoline-3-carboxylate 5a (2.72 g, 8.51 mmol, 32.8 % yield) as a waxy yellow solid. H
NMR (400 MHz, CDCl3) δ 9.25 (s, 1H), 8.48 (d, 1H, J = 9 Hz), 7.98 (dq, 1H, J = 2, 1 Hz), 7.56 (dq, 1H, J = 9, 2 Hz),
4.52 (q, 2H, J = 7 Hz), 1.48 (t, 3H, J = 7 Hz); LC-MS (LC-ES) M+H = 320.
Ethyl 7-(trifluoromethoxy)quinoline-3-carboxylate 6a
O
F
O
F
F
O
N
6a
Bis(triphenylphosphine)palladium(II) dichloride (0.296 g, 0.422 mmol) was added to ethyl 4-chloro-7-
(trifluoromethoxy)quinoline-3-carboxylate 5a (2.70 g, 8.45 mmol) in acetonitrile (24 mL) and the reaction mixture
was degassed for fifteen minutes by sparging with nitrogen. Triethylsilane (1.637 mL, 10.14 mmol) was added via
syringe and the reaction mixture was heated in a sealed tube at 80 °C for nineteen hours. Upon cooling, the volatiles
were removed in vacuo. The residue was taken up in ethyl acetate, filtered through Celite®, washed with saturated
sodium bicarbonate, water, and saturated sodium chloride, dried over sodium sulfate, and concentrated. The residue
was purified by flash chromatography, eluting with ethyl acetate:hexanes (1:49 to 1:4) to afford ethyl 7-
(trifluoromethoxy)quinoline-3-carboxylate 6a (2.075 g, 7.28 mmol, 86 % yield) as a faintly yellow solid. 1H NMR
(400 MHz, CDCl3) δ 9.49 (d, 1H, J = 2 Hz), 8.86 (d, 1H, J = 2 Hz), 8.01 (s, 1H), 8.00 (d, 1H, J = 9 Hz), 7.49 (dd, 1H,
J = 9, 2 Hz), 4.50 (q, 2H, J = 7 Hz), 1.48 (t, 3H, J = 7 Hz); LC-MS (LC-ES) M+H = 286.
7-(Trifluoromethoxy)quinoline-3-carboxylic acid 7c
O
F
OH
F
F
O
N
7c
Lithium hydroxide monohydrate (1.515 g, 36.1 mmol) was added to a solution of ethyl 7-(trifluoromethoxy)quinoline-
3-carboxylate 6a (2.06 g, 7.22 mmol) in tetrahydrofuran (10 mL) and water (2 mL) and the reaction mixture was
stirred at 60 °C for one hour. Upon cooling, the reaction mixture was poured into water (100 mL) and extracted with
diethyl ether (1X). The aqueous layer was acidified by addition of 1 M hydrochloric acid (36 mL) and the precipitated
solids were collected by filtration, washed with water, and air dried on the Buchner funnel. Then, further drying in a
vacuum oven (50°C / 25" Hg) afforded 7-(trifluoromethoxy)quinoline-3-carboxylic acid 7c (1.79 g, 6.96 mmol, 96 %
1
yield) as a colorless solid. H NMR (400 MHz, CD3SOCD3) δ 13.67 (br s, 1H), 9.37 (d, 1H, J = 2 Hz), 9.07 (dd, 1H,
J = 2, 1 Hz), 8.39 (d, 1H, J = 9 Hz), 8.01 (br s, 1H), 7.74 (ddd, 1H, J = 9, 2, 1 Hz); LC-MS (LC-ES) M+H = 258.
(S)-N-(2-Oxopyrrolidin-3-yl)-7-(trifluoromethoxy)quinoline-3-carboxamide 1bq.
O
NH
O
F
N
H
F
F
O
N
1bq
Method B; 7-(Trifluoromethoxy)quinoline-3-carboxylic acid 7c; (S)-3-Amino-2-pyrrolidinone 8ao (Biofine); N,N-
Dimethylformamide; 8% yield; 1H NMR (400 MHz, CD3OD) δ 9.33 (d, 1H, J = 2 Hz), 8.86 (d, 1H, J = 2 Hz), 8.42
(br s, 1H), 8.18 (d, 1H, J = 9 Hz), 7.94 (br s, 1H), 7.62 (dd, 1H, J = 9, 2 Hz), 4.84-4.74 (m, 1H), 3.50-3.38 (m, 2H),
2.66-2.56 (m, 1H), 2.28-2.16 (m, 1H); LC-MS (LC-ES) M+H = 340.
N-(1-(1-methyl-1H-tetrazol-5-yl)piperidin-4-yl)-7-(trifluoromethoxy)quinoline-3-carboxamide 1br.