European Journal of Medicinal Chemistry 89 (2015) 616e627

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European Journal of Medicinal Chemistry

journal homepage: http://www.elsevier.com/locate/ejmech

Original article

Synthesis, molecular docking and anti-mycobacterial evaluation of

new imidazo[1,2-a]pyridine-2-carboxamide derivatives
Gilish Jose a, T.H. Suresha Kumara a, e, *, Gopalpur Nagendrappa a, H.B.V. Sowmya a,
Dharmarajan Sriram b, Perumal Yogeeswari b, Jonnalagadda Padma Sridevi b,
Tayur N. Guru Row c, Amar A. Hosamani c, P.S. Sujan Ganapathy d, N. Chandrika a,
L.V. Narendra a
a
Postgraduate Department of Chemistry, Jain University, 52 Bellary Road, Hebbal, Bangalore, Karnataka 560024, India
b
Department of Pharmacy, Birla Institute of Technology and Science-Pilani, Hyderabad 500078, India
c
Solid State and Structural Chemistry Unit, Indian Institute of Science, Bangalore, Karnataka 560012, India
d
Centre for Advanced Studies in Biosciences, Jain University, 52 Bellary Road, Hebbal, Bangalore, Karnataka 560024, India
e
Department of Chemistry, University B.D.T. College of Engineering, Davangere, Karnataka 577004, India

a r t i c l e i n f o a b s t r a c t

Article history: New anti-tubercular agents, imidazo[1,2-a]pyridine-2-carboxamide derivatives (5aeq) have been
Received 23 June 2014 designed and synthesized. The structural considerations of the designed molecules were further sup-
Received in revised form ported by the docking study with a long-chain enoyl-acyl carrier protein reductase (InhA). The chemical
8 October 2014
structures of the new compounds were characterized by IR, 1H NMR, 13C NMR, HRMS and elemental
Accepted 29 October 2014
Available online 30 October 2014
analysis. In addition, single crystal X-ray diffraction has also been recorded for compound 5f. Compounds
were evaluated in vitro against Mycobacterium tuberculosis H37Rv, and cytotoxicity against HEK-293T cell
line. Amongst the tested compounds 5j, 5l and 5q were emerged as good anti-tubercular agents with low
Keywords:
Anti-mycobacterial activity
cytotoxicity. The structure-anti TB activity relationship of these derivatives was explained by molecular
Imidazopyridine amide docking.
InhA © 2014 Elsevier Masson SAS. All rights reserved.
Tuberculosis
X-ray crystallography

1. Introduction accounts for 26% of AIDS related death worldwide [3e7]. In 2012,
an estimated 8.6 million people affected by M. tuberculosis and 1.3
TB is a chronic bacterial infection is often contracted through an million died from the disease, including 320,000 deaths among
airborne bacterium known as Mycobacterium tuberculosis. HIV-positive people [8]. To address these issues, design, synthesize
Although most M. tuberculosis infections, known as pulmonary TB, and develop potent anti-mycobacterial agents are necessary.
are in the lungs, 5 to 10 percent of TB patients can develop the Imidazopyridine derivatives are very important, versatile motifs
disease in organs such as lymph nodes, bones and joints, eyes, in- with significant applications in medicinal chemistry [9e14].
testines, larynx, or the urinary and reproductive systems, skin, and Different compounds containing the moiety imidazo[1,2-a]pyridine
stomach, known as extrapulmonary TB [1,2]. In recent years, the have significant biological applications such as anti-mycobacterial
emergence of MDR and XDR tuberculosis strains has amplified the [15e21], anticancer [22,23], antiviral [24e27], antimicrobial
incidences of TB. In addition, TB is a co-infection of HIV-AIDS and [28e30], anti-HIV [31], antiulcer [32,33], antihelminthic [34] and
anticoccidial activity [35,36]. As a result of our interest in the
synthesis of imidazopyridine-containing compounds and their
Abbreviations: TB, tuberculosis; MDR, multiple drug-resistant; XDR, extensively antimicrobial activity [37], we initiated a program directed towards
drug-resistant; HIV-AIDS, human immunodeficiency virus-acquired immunodefi- the synthesis of the new imidazopyridine amide derivatives. Imi-
ciency syndrome; DIPEA, diisopropylethylamine; HATU, 1-[bis(dimethylamino)
dazopyridine amides (IPA) are a new class of therapeutic agents
methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxid hexafluorophosphate; MIC,
minimum inhibitory concentration. against MDR and XDR tuberculosis [18e21]. Recently, Qurient
* Corresponding author. Department of Chemistry, University B.D.T. College of Therapeutics discovered Q203 (Fig. 1), a highly innovative drug for
Engineering, Davangere, Karnataka 577004, India. MDR and XDR tuberculosis and the first drug in its class with an
E-mail addresses: [email protected], [email protected] original mechanism of action [21]. On the basis of above beneficial
(T.H. Suresha Kumara).

http://dx.doi.org/10.1016/j.ejmech.2014.10.079
0223-5234/© 2014 Elsevier Masson SAS. All rights reserved.
 G. Jose et al. / European Journal of Medicinal Chemistry 89 (2015) 616e627 617

information, we earmarked to design new imidazo[1,2-a]pyridine- were synthesized by using the same experimental protocol with
2-carboxamide derivatives anticipating enhanced biological activ- different aromatic or aliphatic amines (R1eNH2). The Suzuki re-
ity to combat this lethal disease. The structural considerations of action of 4a with 1-methylpyrazole-4-boronic acid pinacol ester,
the designed molecules were further supported by the docking catalysed by Pd(0) complex in the presence of Na2CO3 gave final
study with target enzyme InhA, an enzyme essential for mycolic product 5a. IR spectrum of 4a showed that the appearance of the
acid biosynthesis in M. tuberculosis. The new target compounds band at 1650 cm
1 characteristic for amide C]O stretch. 1H NMR
were designed based on the assumptions of Lipinski rule to fulfil and 13C NMR spectra of 5a were revealed that the presence of the
the drug likeness properties of the molecules. We report herein, carboxamide group at position 2, showed a triplet at d 8.93 ppm
synthesis, crystal structure, molecular docking and anti-mycobac- and d 162.3 ppm, respectively. The band at 1652 cm
1 in the IR
terial evaluation of new imidazo[1,2-a]pyridine-2-carboxamide spectrum of 5a characteristic for the amide C]O stretch. The
derivatives. HRMS spectrum of 5a showed 350.1405 [MþH]þ corresponding
with proposed isotopic mass (349.1338). The obtained elemental
analysis values are in consonance with theoretical data. The
2. Results and discussion
remaining final compounds (5beq) were synthesized using the
same experimental procedure with different aromatic or hetero
2.1. Chemistry
aromatic boronic acids (R2). The structures of the final products
were assigned on the basis of their IR, 1H NMR, 13C NMR, HRMS
The synthetic strategy of new polyfunctional imidazo[1,2-a]
and elemental analysis. The structure of the compound 5f (Fig. 2)
pyridine-2-carboxamides were described in Scheme 1. The imi-
was unequivocally established by X-ray crystallographic analysis.
dazo[1,2-a]pyridine ring system was accomplished by the
The details of the crystal data and structure refinement was
condensation of 2-amino-5-iodopyridine (1) with ethyl bromo-
shown in Table 1.
pyruvate (a-halocarbonyl compound) in refluxing ethanol to gave
ethyl-6-iodo-H-imidazo[1,2-a]pyridine-2-carboxylate (2) [38]. 1H
NMR spectrum of 2 showed a triplet at d 1.30 (3H) and a quartet 2.2. Pharmacology
at d 4.30 (2H) ppm corresponds to an ethyl ester group at position
2. 13C NMR spectrum of 2 showed peaks at d 167.3 All the synthesized compounds were also screened for their
(COOeCH2eCH3), 60.3 (OeCH2), 14.8 (CH3) ppm also revealed in vitro anti-tubercular activity against M. tuberculosis H37Rv
that the presence of ethyl ester group. IR spectrum of 2 revealed (ATCC 27294) using microplate alamar blue assay (MABA) with
that appearance of the band at 1748 cm
1 characteristic for ester drug concentrations from 50 mg/mL to 0.78 mg/mL in duplicates.
C]O stretch. The ester hydrolysis reaction of 2 with lithium The MIC was determined for each compound which was
hydroxide monohydrate gave 6-iodo-H-imidazo[1,2-a]pyridine- measured as the minimum concentration of compound required
2-carboxylic acid (3). 1H NMR and 13C NMR spectra of 3 exhibited to completely inhibit the bacterial growth. Isoniazid and rifam-
a broad singlet peak at d 11.50 ppm and a peak at d 167.1 ppm picin were used as reference compounds for comparison. The
were assigned for carboxylic acid, while its IR spectrum showed a in vitro test results for title compounds were tabulated in Table 2
strong band at 1755 cm
1 also revealed that the presence of as the MIC and the activity ranged from 6.25 to >50 mg/mL.
carboxylic acid. The peptide coupling reaction between 3 and 4- Various substitutions were attempted at R1 and R2 position of the
fluorobenzylamine was accomplished by famous peptide imidazo[1,2-a]pyridine core as steps towards the derivation of
coupling reagent, HATU and Hunig's base (DIPEA) in DMF gave structureeactivity relationship and to understand the ideal site
scaffold 4a. IR spectrum of 4a showed that the appearance of the for introducing chemical diversity. Out of the various compounds
band at 1650 cm
1 characteristic for amide C]O stretch. 1H NMR tested, compounds 5h, 5j, 5k, 5l, 5o and 5q inhibited mycobac-
spectrum of 4a showed a triplet at d 9.00 ppm revealed that the terial growth very effectively compared to others in the series
presence of the carboxamide group at position 2, while its 13C with MIC values ranging from 6.25 to 12.5 mg/mL. The highest
NMR spectrum showed d 162.4 ppm also revealed that the activity, 6.25 mg/mL was registered for compounds 5j
presence of carboxamide group. The remaining scaffolds (4beg) (R1 ¼ pyridin-4-yl methyl, R2 ¼ 4-chlorophenyl), 5l (R1 ¼ benzo

Fig. 1. Some of the imidazopyridine amide based anti-tubercular agents.

618 G. Jose et al. / European Journal of Medicinal Chemistry 89 (2015) 616e627

Scheme 1. Synthesis of imidazo[1,2-a]pyridine-2-carboxamide derivatives; Reagents and conditions: (i) ethyl alcohol, reflux, 4 h; (ii) LiOH, methanol/THF/H2O, 60  C, 1 h; (iii)
R1eNH2, DIPEA, HATU, DMF, rt, 12 h (iv) R2eB(OH)2, Na2CO3, Pd(PPh3)4, 1,4-dioxane/H2O, 90  C, 8 h.

[d][1,3]dioxol-5-yl methyl, R2 ¼ 1-methyl-1H-pyrazol-4-yl) and 2.3. Molecular docking studies

5q (R1 ¼ tetrahydro-2H-pyran-4-yl, R2 ¼ 1-(2-morpholinoethyl)-
1H-pyrazol-4-yl). It was observed that the presence of the hy- The molecular docking of imidazo[1,2-a]pyridine carboxamide
drophobic substituents on both R1 and R2 position of the imidazo motifs (5aeq) with a long-chain enoyl-acyl carrier protein reduc-
[1,2-a]pyridine core brought about an enhancement of the anti- tase (InhA) yielded best possible conformations with parameters
mycobacterial potency as the overall hydrophobic nature might including the docking energy, binding energy, inhibition constant
have enabled these molecules to penetrate the quite complex and intermolecular energy (Table 3). The docking results are drawn
mycobacterial cell wall, a major hurdle faced in anti-tubercular based on both binding energy and the number of hydrogen bonds
drug discovery. Lipophilicities of the compounds 5aeq and the formed. The intermolecular hydrogen bond interactions play an
standard drugs, which were expressed as logP values, were important role in adduct binding in InhA, especially the hydrogen
determined using the logP method through online http://www. bonds of the pyrophosphate part. The main residues involved in
molinspiration.com/cgi-bin/properties site, as shown in Table 2. these interactions are His 265, Tyr 259, Trp 249, Gly 3, Ile 194, and
The safety profile of the active leads was also accessed by testing Arg 173. Compounds (5aeq) showed very good docking energy
in vitro cytotoxicity against Human Embryonic Kidney Cell-line ranging from
12.07 kJ/mol to
9.42 kJ/mol because target com-
293T (HEK-293T) cells at 50 mg/mL concentration by (4,5- pounds have formed one or more hydrogen bonds with amino acid
dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) residues in the active pocket of InhA (Fig. 3). Compounds contain-
assay. Percentage inhibitions of cells are reported in Table 2. The ing the halogen atoms (5aed, 5j, 5n, 5o) also have the ability to
entire tested compound demonstrated a good safety profile with form intermolecular bonds with the active site of InhA in a fashion
very low toxicity towards the HEK cells and showed a good selec- that resembles hydrogen bonds. All compounds, except 5c, 5g and
tivity index (CC50/MIC) indicating the suitability of these com- 5n have formed two or more hydrogen bonds with amino acid
pounds for further drug development. residues of InhA. The maximum number of hydrogen bonds was
found in the analogue 5o (3 hydrogen bonds) with a binding energy
(
9.20 kJ mol
1), docking energy (
11.62 kJ mol
1) and inhibition
constant (1.79  10
7 M). In in vitro anti-mycobacterial studies,
compounds 5h, 5j, 5k, 5l, 5o and 5q have emerged as active against
microbe, so it can be predicted as the activity may be due to inhi-
bition of enzyme InhA.

3. Conclusions

In conclusion, new anti-tubercular agents imidazo [1,2-a] pyri-

dine-2-carboxamide derivatives 5aeq was efficiently synthesized.
The activity of these compounds against M. tuberculosis strain
H37Rv was assessed and compounds 5j, 5l and 5q were displayed
good anti-tubercular activity. Their activity depends on both the
substituent's R1 and R2. The in vitro cytotoxicity study against HEK-
293T cells at 50 mg/mL by MTT assay revealed that the identified hits
are found to be less cytotoxic. The molecular docking of InhA
domain with synthesized compounds revealed that compounds 5h,
5j, 5k, 5l, 5o and 5q having the good inhibitory capability and are
exhibiting the interactions with one or the other amino acids in the
Fig. 2. X-ray crystal structure of 5f. active pocket. With new antiTB agents desperately needed, we
 G. Jose et al. / European Journal of Medicinal Chemistry 89 (2015) 616e627 619

Table 1 reduced pressure. The residue was neutralized with 10% NaHCO3
Crystal data and structure refinement for 5f. solution (250 mL). The solid formed was collected by filtration,
Identification code 5f washed with water and dried. Off white solid (26.7 g, 94%); mp
Empirical formula C18H15N5O2 135.8e138.6  C; IR (KBr, nmax, cm
1): 3163 (CeHarom.), 2918
Formula weight 333.344 (CeHaliph.), 1748 (C]Oester), 1575 (CeCarom.); 1H NMR (400 MHz,
Temperature/K 296.15
Crystal system Monoclinic
DMSO-D6): d 8.94 (s, 1H, H-5), 8.43 (s, 1H, H-3), 7.53 (d, J ¼ 9.6 Hz,
Space group P21/c 1H, H-7), 7.47 (d, J ¼ 9.6 Hz, 1H, H-8), 4.30 (q, J ¼ 7.2 Hz, 2H,
a/Å 31.097(6) OeCH2eCH3), 1.30 (t, J ¼ 7.2 Hz, 3H, CH2eCH3); 13C NMR (100 MHz,
b/Å 9.3652(19) DMSO-D6): d 167.3 (COOeCH2eCH3), 156.4 (C-5), 149.3 (C-9), 144.5
c/Å 10.816(2)
(C-7), 138.8 (C-2), 118.9 (C-3), 118.1 (C-8), 96.2 (C-6), 60.3 (OeCH2),
a/ 90.00
b/ 90.084(11) 14.8 (CH3); LC/MS (API-ES) m/z calcd. for C10H9IN2O2: 315.971,
g/ 90.00 found: 317.0 [MþH]þ; Anal. calcd. for C10H9IN2O2: C, 38.00; H, 2.87;
Volume/Å3 3149.9(11) N, 8.86; found C, 37.98; H, 2.859; N, 8.81.
Z 4
rcalc mg/mm3 1.383
m/mm
1 0.090
4.3. Experimental procedure for the synthesis of 6-iodoH-imidazo
F(000) 1368.0 [1,2-a]pyridine-2-carboxylic acid (3)
Crystal size/mm3 0.21  0.18  0.26
Radiation MoKa (l ¼ 0.71073) A mixture of compound 2 (26.7 g, 84.46 mmol) and lithium
2Q range for data collection 2.62e50
hydroxide monohydrate (10.63 g, 253.38 mmol) and 270 mL of
Index ranges
36  h  36,
11  k  10,
12  l  12
Reflections collected 35,922 methanol/THF/water (7:2:1) were heated at 60  C for 1 h. The
Independent reflections 5523 [Rint ¼ 0.1491, Rsigma ¼ 0.1080] solvent was concentrated under reduced pressure. The residue was
Data/restraints/parameters 5523/0/454 dissolved in 80 mL of water. The PH of the reaction mixture was
Goodness-of-fit on F2 1.016 adjusted to 6 by using 10% citric acid solution. The solid formed was
Final R indexes [I  2s (I)] R1 ¼ 0.0908, wR2 ¼ 0.2394
Final R indexes [all data] R1 ¼ 0.1799, wR2 ¼ 0.2817
collected by filtration, washed with water and dried. White solid
Largest diff. peak/hole/e Å
3 0.44/
0.60 (22.6 g, 93%); mp 185.9e188.4  C; IR (KBr, nmax, cm
1): 3166
(CeHarom.), 2520 (OeHacid), 2920 (CeHaliph.), 1755 (C]Oacid), 1576
(CeCarom.), 1221 (CeOacid); 1H NMR (400 MHz, DMSO-D6): d 11.50
believe that the new imidazopyridines reported in this work pro- (br s, 1H, COOH), 8.94 (s, 1H, H-5), 8.30 (s, 1H, H-3), 7.48 (d,
vide an interesting potential for further optimization. Further J ¼ 9.6 Hz, 1H, H-7), 7.43 (d, J ¼ 9.6 Hz, 1H, H-8); 13C NMR (100 MHz,
structural modifications of the identified hits are in progress in DMSO-D6): d 167.1 (COOH), 156.4 (C-5), 149.4 (C-9), 144.5 (C-7),
order to enhance the efficacy of these compounds active against 138.9 (C-2), 118.8 (C-3), 118.2 (C-8), 96.1 (C-6); LC/MS (API-ES) m/z
M. tuberculosis. calcd. for C8H5IN2O: 287.940, found: 287.0 [M-H]þ; Anal. calcd. for
C8H5IN2O2: C, 33.36; H, 1.75; N, 9.73; found: C, 33.32; H, 1.758; N,
9.69.
4. Experimental section
4.4. General experimental procedure for the synthesis of 4aeg
4.1. General methods
Synthesis of N-(4-fluorobenzyl)-6-iodo1H-imidazo[1,2-a]pyridine-
All chemicals and solvents were purchased from commercial 2-carboxamide (4a) as an example: To a mixture of compound 3 (1 g,
suppliers. All reactions were performed under an inert atmosphere 3.47 mmol), 4-fluorobenzylamine (0.65 g, 5.21 mmol) and DIPEA
of dry nitrogen using dry solvents. The progress of the reactions (2.4 mL, 13.89 mmol) in DMF (10 mL), HATU (1.98 g, 5.21 mmol) was
was monitored by using Si gel 60 F254 thin layer chromatography added portion wise at 0  C. The resulting reaction mixture was
plates and spots were detected by UV/254 nm. Compounds were stirred at rt for 12 h. The solvent was removed under reduced
purified by column chromatography on silica gel 230e400 using pressure. The reaction mixture was dissolved in DCM (50 mL) and
petether/ethyl acetate or dichloromethane/methanol solvent mix- the organic layer was washed with water (25 mL) and brine
tures as eluent. All the compounds were characterized by IR, 1H (25 mL). The organic layer was separated and dried over anhydrous
NMR, 13C NMR, HRMS and elemental analysis. Melting points of the MgSO4. The organic layer was concentrated in vacuum. The crude
samples were determined in open capillary tubes using Buchi product was purified via flash column chromatography on silica gel
Melting point B-540 apparatus and are uncorrected. IR spectra were (230e400 mesh) to provide 4a as pale yellow solid (1.27 g, 93%).
obtained on a PerkinElmer Frontier™ FT-IR spectrometer, were
recorded in KBr. 1H and 13C NMR spectra were recorded on a Bruker 4.4.1. N-(4-Fluorobenzyl)-6-iodo1H-imidazo[1,2-a]pyridine-2-
NMR spectrometer (400 and 100 MHz, respectively) using deuter- carboxamide (4a)
ated DMSO as solvent with TMS as internal standard. The mass mp 195.1e197.0  C; IR (KBr, nmax, cm
1): 3354 (NeHamide), 3163,
recorded in Agilent LCMS 1100 series instrument using API-ES 3076 (CeHarom.), 2918 (CeHaliph.), 1650 (C]Oamide), 1574
positive-ion mode and negative-ion mode. HRMS was recorded in (CeCarom.); 1H NMR (300 MHz, DMSO-D6): d 9.00 (t, J ¼ 6.0 Hz, 1H,
positive-ion mode by Microflex LT MALDI-ToF mass spectrometer COeNHeCH2), 8.95 (s, 1H, H-5), 8.26 (s, 1H, H-3), 7.46 (d,
(Bruker Daltonics, Germany). The CHN analysis was recorded in an J ¼ 10.4 Hz, 2H, H-20 , H-60 ), 7.36e7.32 (m, 2H, AreH), 7.11 (d,
Elementar vario MICRO cube. J ¼ 12.0 Hz, 2H, H-30 , H-50 ), 4.41 (d, J ¼ 8.4 Hz, 2H,
COeNHeCH2eAr); 13C NMR (100 MHz, DMSO-D6): d 162.4
4.2. Experimental procedure for the synthesis of ethyl 6-iodoH- (COeNH), 162.1 (C-40 ), 159.7 (C-9), 156.3 (C-5), 144.5 (C-7), 136.0 (C-
imidazo[1,2-a]pyridine-2-carboxylate (2) 10 ), 135.9 (C-2), 129.4 (C-20 ), 129.3 (C-60 ), 118.8 (C-3), 118.2 (C-8),
114.9 (C-30 ), 114.7 (C-50 ), 96.2 (C-6), 41.2 (COeNHeCH2eAr); LC/MS
A mixture of 2-amino-5-iodo pyridine 1 (20 g, 90.90 mmol) and (API-ES) m/z calcd. for C15H11FIN3O: 394.993, found: 396.1 [MþH]þ;
ethyl bromopyruate (53.17 g, 272.70 mmol) were refluxed in Anal. calcd. for C15H11FIN3O: C, 45.59; H, 2.81; N, 10.63, found: C,
200 mL of ethanol for 4 h. The solvent was concentrated under 45.56; H, 2.808; N, 10.60.
620 G. Jose et al. / European Journal of Medicinal Chemistry 89 (2015) 616e627

Table 2
Anti-mycobacterial activity and cytotoxicity of compounds 5aeq.

Compound R1 R2 MIC (mg/mL)a Cytotoxicity at 50 mg/mLb LogP

5a 25 36.12 2.420

5b 25 40.8 2.578

5c 25 28.12 4.302

5d >50 NT 3.985

5e >50 NT 1.513

5f 25 26.12 1.672

5g 25 24.8 3.396

5h 12.5 34.06 1.389

5i 25 30.12 1.994

5j 6.25 26.12 3.268

5k 12.5 31.62 2.598

 G. Jose et al. / European Journal of Medicinal Chemistry 89 (2015) 616e627 621

Table 2 (continued )

Compound R1 R2 MIC (mg/mL)a Cytotoxicity at 50 mg/mLb LogP

5l 6.25 25.56 2.146

5m 25 29.12 1.099

5n 25 37.80 5.034

5o 12.5 22.16 3.027

5p 25 30.7 1.232

5q 6.25 30.06 3.114

Isoniazid e e 0.36 e
0.70

Rifampicin e e 0.02 e 2.770
a
Anti-mycobacterial activity against M. tuberculosis strain H37Rv.
b
Percentage of cell inhibition at 50 mg/mL against HEK-293T cells; NT denotes not tested.

4.4.2. N-(Furan-2-ylmethyl)-6-iodo1H-imidazo[1,2-a]pyridine-2- 60 ), 144.5 (C-7), 139.9 (C-40 ), 135.1 (C-2), 122.2 (C-30 , C-50 ), 118.6 (C-
carboxamide (4b) 3), 118.2 (C-8), 96.3 (C-6), 41.1 (COeNHeCH2eAr); LC/MS (API-ES)
Prepared from 3 (1 g, 3.47 mmol), furfurylamine (0.51 g, m/z calcd. for C14H11IN4O: 377.998, found: 379.1 [MþH]þ; Anal.
5.21 mmol), DIPEA (2.4 mL,13.89 mmol), HATU (1.98 g, 5.21 mmol) in calcd. for C14H11IN4O: C, 44.46; H, 2.93; N, 14.82; found: C, 44.44; H,
DMF (10 mL). 4b was formed as a pale brown crystal (1.11 g, 87%); mp 2.925; N, 14.83.
201.2e203.0  C; 1H NMR (400 MHz, DMSO-D6): d 8.96 (s, 1H, H-5),
8.78 (t, J ¼ 6.0 Hz, 1H, COeNHeCH2), 8.28 (s, 1H, H-3), 7.55e7.50 (m, 4.4.4. N-(Benzo[d][1,3]dioxol-5-ylmethyl)-6-iodoH-imidazo[1,2-a]
2H, AreH), 7.43 (d, J ¼ 9.6 Hz, 1H, H-50 ), 6.36 (t, J ¼ 3.2 Hz, 1H, H-40 ), pyridine-2-carboxamide (4d)
6.22 (d, J ¼ 3.1 Hz,1H, H-30 ), 4.43 (d, J ¼ 6.1 Hz, 2H, COeNHeCH2eAr); Prepared from 3 (1 g, 3.47 mmol), piperonylamine (0.79 g,
13
C NMR (100 MHz, DMSO-D6): d 161.8 (COeNH), 156.3 (C-5), 152.5 5.21 mmol), DIPEA (2.4 mL, 13.89 mmol), HATU (1.98 g, 5.21 mmol)
(C-9), 144.5 (C-7), 141.8 (C-20 ), 139.4 (C-50 ), 135.9 (C-2), 119.0 (C-3), in DMF (10 mL). 4d was formed as an off white solid (1.34 g, 92%);
118.2 (C-8), 110.4 (C-40 ), 106.6 (C-30 ), 96.2 (C-6), 35.3 mp 211.0e213.2  C; 1H NMR (300 MHz, DMSO-D6): d 8.95 (s, 1H, H-
(COeNHeCH2eAr); LC/MS (API-ES) m/z calcd. for [C13H10IN3O2]: 5), 8.88 (t, J ¼ 6.4 Hz, 1H, COeNHeCH2), 8.26 (s, 1H, H-3), 7.49 (d,
366.982, found: 368.0 [MþH]þ; Anal. calcd. for C13H10IN3O2: C, J ¼ 12.0 Hz, 1H, H-7), 7.41 (d, J ¼ 12.0 Hz, 1H, H-8), 6.88 (s, 1H, H-40 ),
42.53; H, 2.75; N, 11.45; found: C, 42.56; H, 2.759; N, 11.43. 6.83e6.76 (m, 2H, AreH), 5.94 (s, 2H, OeCH2eO), 4.33 (d, J ¼ 5.8 Hz,
2H, COeNHeCH2eAr); 13C NMR (100 MHz, DMSO-D6): d 161.9
4.4.3. 6-Iodo-N-(pyridin-4-ylmethyl)H-imidazo[1,2-a]pyridine-2- (COeNH), 156.5 (C-5), 147.1 (C-9), 145.9 (H-3a0 ), 144.5 (C-7), 135.9
carboxamide (4c) (C-2), 133.7 (H-7a0 ), 120.6 (C-50 ), 118.8 (C-3), 117.8 (C-60 ), 118.2 (C-8),
Prepared from 3 (1 g, 3.47 mmol), 4-(aminomethyl)pyridine 108.1 (C-70 ), 107.9 (C-40 ), 100.7 (OeCH2eO), 96.3 (C-6), 41.7
(0.56 g, 5.21 mmol), DIPEA (2.4 mL, 13.89 mmol), HATU (1.98 g, (COeNHeCH2eAr); LC/MS (API-ES) m/z calcd. for C16H12IN3O3:
5.21 mmol) in DMF (10 mL). 4c was formed as an off white solid 420.992, found: 422.0 [MþH]þ; Anal. calcd. for C16H12IN3O3: C,
(1.17 g, 89%); mp 165.8e168.3  C; 1H NMR (400 MHz, DMSO-D6): 45.63; H, 2.87; N, 9.98; found: C, 45.60; H, 2.871; N, 10.00.
d 9.13 (t, J ¼ 6.4 Hz, 1H, COeNHeCH2), 8.97 (s, 1H, H-5), 8.48 (d,
J ¼ 6.0 Hz, 2H, H-20 , H-60 ), 8.29 (s, 1H, H-3), 7.53 (d, J ¼ 11.1 Hz, 1H, 4.4.5. N-Cyclopropyl-6-iodoH-imidazo[1,2-a]pyridine-2-
H-7), 7.45 (d, J ¼ 9.5 Hz, 1H, H-8), 7.29 (d, J ¼ 5.8 Hz, 2H, H-30 , H-50 ), carboxamide (4e)
4.47 (d, J ¼ 6.3 Hz, 1H, COeNHeCH2eAr); 13C NMR (100 MHz, Prepared from 3 (1 g, 3.47 mmol), cyclopropylamine (0.3 g,
DMSO-D6): d 162.3 (COeNH), 156.4 (C-5), 149.4 (C-9), 148.7 (C-20 , C- 5.21 mmol), DIPEA (2.4 mL, 13.89 mmol), HATU (1.98 g, 5.21 mmol)
622 G. Jose et al. / European Journal of Medicinal Chemistry 89 (2015) 616e627

Table 3
Molecular docking of imidazo[1,2-a]pyridine-2-carboxamide motifs (5aeq) with InhA.

Compound Binding energy (kJ/mol) Docking energy (kJ/mol) Inhibition constant (M) Intermolecular energy (kJ/mol) H-bonds Bonding

5a
8.31
9.42 8.12  10
7
9.55 2 5a::DRG1:HAZ:INHA:B:HIS265:O

5a::DRG1:NAM:INHA:A:TYR259:HH
5b
9.64
11.16 8.60  10
8
10.88 2 5b::DRG1:HBA:INHA:A:GLN32:OE1
5b::DRG1:OAM:INHA:A:TRP249:HE1

8
5c
10.56
12.07 1.81  10
11.81 1 5c::DRG1:OAM:INHA:B:ILE194:HN
5d
9.85
11.86 6.02  10
8
12.03 2 5d::DRG1:OBE:INHA:A:ILE21:HN
5d::DRG1:OBD:INHA:A:ALA22:HN
5e
8.75
10.16 3.83  10
7
10.00 2 5e::DRG1:HAY:INHA:B:GLY3:O
5e::DRG1:OAR:INHA:B:TRP249:HE1
5f
9.46
10.95 1.16  10
7
10.71 2 5f::DRG1:HAZ:INHA:B:ILE194:O
5f::DRG1:NAQ:INHA:B:ILE194:HN

8
5g
9.66
11.16 8.34  10
10.90 1 5g::DRG1:OAM:INHA:B:ILE194:HN
5h
8.72
10.33 4.06  10
7
10.90 2 5h::DRG1:OAU:INHA:A:LYS8:HZ2
5h::DRG1:OBC:INHA:B:ASN231:HN
5i
8.66
10.59 4.51  10
7
10.53 2 5i::DRG1:NAI:INHA:B:ARG153:HH11
5i::DRG1:OAL:INHA:B:HIS265:HD1
5j
9.00
10.08 2.53  10
7
10.25 2 5j::DRG1:NAR:INHA:B:MET98:HN
5j::DRG1:OAM:INHA:B:TYR158:HH

7
5k
8.55
10.18 5.38  10
10.11 2 5k::DRG1:NAR:INHA:A:TRP249:HE1
5k::DRG1:NAI:INHA:B:TRP249:HE1

7
5l
8.85
9.94 3.25  10
10.1 2 5l::DRG1:HBC:INHA:B:HIS265:O
5l::DRG1:NAM:INHA:B:ARG225:HH11
5m
9.04
10.28 2.37  10
7
10.91 2 5m::DRG1:HBE:INHA:A:ILE257:O
5m::DRG1:OAR:INHA:B:ARG173:HH12
5n
9.53
10.95 1.03  10
7
11.09 1 5n::DRG1:OAM:INHA:A:TRP249:HE1
5o
9.20
11.62 1.79  10
7
11.69 3 5o::DRG1:HAQ:INHA:A:ASP256:OD1
5o::DRG1:NAM:INHA:B:ARG173:HH21
5o::DRG1:OAR:INHA:A:TYR259:HH
5p
9.43
10.09 1.23  10
7
10.36 2 5p::DRG1:NAM:INHA:A:THR2:HN1
5p::DRG1:OAR:INHA:B:TRP249:HE1
5q
10.03
10.61 4.44  10
8
10.96 2 5q::DRG1:HAL:INHA:B:GLY3:O
5q::DRG1:OAM:INHA:B:TRP249:HE1

in DMF (10 mL). 4e was formed as an off white solid (0.96 g, 85%); (1.11 g, 86%); mp 111.3e113.6  C; 1H NMR (300 MHz, DMSO-D6):
mp 125.5e128.3  C; 1H NMR (300 MHz, DMSO-D6): d 8.95 (s, 1H, H- d 8.95 (s, 1H, H-5), 8.27 (d, J ¼ 8.4 Hz, 1H, COeNHeCH), 8.25 (s, 1H,
5), 8.38 (d, J ¼ 4.7 Hz, 1H, COeNHeCH), 8.24 (s, 1H, H-3), 7.49 (d, H-3), 7.49 (d, J ¼ 9.3 Hz, 1H, H-7), 7.41 (d, J ¼ 9.3 Hz, 1H, H-8),
J ¼ 12.0 Hz, 1H, H-7), 7.39 (d, J ¼ 9.4 Hz, 1H, H-8), 2.86e2.82 (m, 1H, 4.02e3.95 (m, 1H, H-40 ), 3.84 (d, J ¼ 11.2 Hz, 2H, H-2a0 , H-6a0 ),
C-10 ), 0.65e0.50 (m, 4H, H-20 , H-30 ); 13C NMR (100 MHz, DMSO-D6): 3.41e3.35 (dt, J ¼ 11.2, 4.0 Hz, 2H, H-2b0 , H-6b0 ), 1.74e1.66 (m, 4H,
d 163.2 (COeNH), 156.5 (C-5), 144.6 (C-9), 144.4 (C-7), 138.3 (C-2), H-30 , H-50 ); 13C NMR (100 MHz, DMSO-D6): d 161.2 (COeNH), 156.4
125.8 (C-3), 118.2 (C-8), 96.3 (C-6), 24.9 (C-10 ), 5.7 (C-20 ), 5.5 (C-30 ); (C-5), 147.9 (C-9), 144.5 (C-7), 135.9 (C-2), 118.8 (C-3), 118.2 (C-8),
LC/MS (API-ES) m/z calcd. for C11H10IN3O: 326.987, found: 328.0 96.5 (C-6), 66.1 (C-20 , C-60 ), 44.9 (C-40 ), 32.3 (C-30 , C-50 ); LC/MS (API-
[MþH]þ; Anal. calcd. for C11H10IN3O: C, 40.39; H, 3.08; N, 12.85; ES) m/z calcd. for C13H14IN3O2: 371.013, found: 372.2 [MþH]þ; Anal.
found: C, 40.41; H, 3.074; N, 12.85. calcd. for C13H14IN3O2: C, 42.07; H, 3.80; N, 11.32; found C, 42.09; H,
3.809; N, 11.30.
4.4.6. N-(4-(Trifluoromethyl)benzyl)-6-iodoH-imidazo[1,2-a]
pyridine-2-carboxamide (4f) 4.5. General experimental procedure for the synthesis of 5aeq
Prepared from 3 (1 g, 3.47 mmol), 4-(trifluoromethyl)benzyl-
amine (0.91 g, 5.21 mmol), DIPEA (2.4 mL, 13.89 mmol), HATU 4.5.1. Synthesis of N-(4-fluorobenzyl)-6-(1-methyl-1H-pyrazol-4-
(1.98 g, 5.21 mmol) in DMF (10 mL). 4f was formed as a pale yellow yl)H-imidazo[1,2-a]pyridine-2-carboxamide (5a) as an example
solid (1.44 g, 93%); mp 200.2e202.4  C; 1H NMR (300 MHz, DMSO- An oven-dried Schlenk tube was charged with compound 4a
D6): d 9.13 (t, J ¼ 6.0 Hz, 1H, COeNHeCH2), 8.96 (s, 1H, H-5), 8.28 (s, (300 mg, 0.76 mmol), 1-methylpyrazole-4-boronic acid pinacol
1H, H-3), 7.66 (d, J ¼ 8.1 Hz, 2H, H-30 , H-50 ), 7.51 (d, J ¼ 8.4 Hz, 2H, H- ester (236.9 mg, 1.14 mmol), Na2CO3 (241.4 mg, 2.28 mmol) and
20 , H-60 ), 7.49e7.39 (m, 2H, AreH), 4.52 (d, J ¼ 6.2 Hz, 2H, 9.5:0.5 mixture of 1,4-dioxane/water (10 mL). The Schlenk tube was
COeNHeCH2eAr); 13C NMR (100 MHz, DMSO-D6): d 162.3 capped with a rubber septum, evacuated and backfilled with argon
(COeNH), 156.5 (C-5), 144.7 (C-9), 144.4 (C-7), 135.4 (C-2), 133.5 (C- (this sequence was carried out four times). Tetrakis(-
10 ), 127.8 (C-40 ), 127.5 (C-20 , C-60 ), 125.1 (C-30 , C-50 ), 125.0 (AreCF3), triphenylphosphine)palladium(0) (43.9 mg, 0.04 mmol) was added
118.8 (C-3), 118.2 (C-8), 96.4 (C-6), 41.7 (COeNHeCH2eAr); LC/MS and the Schlenk tube was sealed with a Teflon screw cap. The re-
(API-ES) m/z calcd. for C16H11F3IN3O: 444.99, found 446.0 [MþH]þ; action mixture was heated to 90  C for 8 h. The reaction mixture
Anal. calcd. for C16H11F3IN3O: C, 43.17; H, 2.49; N, 9.44; found: C, was filtered through a thin pad of celite (eluted with CH2Cl2) and
43.20; H, 2.504; N, 9.46. the eluent was concentrated under reduced pressure. The crude
product was purified via flash column chromatography on silica gel
4.4.7. 6-Iodo-N-(tetrahydro-2H-pyran-4-yl)H-imidazo[1,2-a] (230e400 mesh) to provide 5a as a white solid (225.4 mg, 85%).
pyridine-2-carboxamide (4g)
Prepared from 3 (1 g, 3.47 mmol), 4-aminotetrahydropyran 4.5.1.1. N-(4-Fluorobenzyl)-6-(1-methyl-1H-pyrazol-4-yl)H-imidazo
(0.53 g, 5.21 mmol), DIPEA (2.4 mL, 13.89 mmol), HATU (1.98 g, [1,2-a]pyridine-2-carboxamide (5a). mp 220.3e222.6  C; IR (KBr,
5.21 mmol) in DMF (10 mL). 4g was formed as an off white solid nmax, cm
1): 3355 (NeHamide), 3163, 3076 (CeHarom.), 2919
 G. Jose et al. / European Journal of Medicinal Chemistry 89 (2015) 616e627 623

Fig. 3. Interaction of compounds 5h, 5j, 5k, 5l, 5o and 5q in the active pocket of InhA.

(CeHaliph.), 1652 (C]Oamide), 1574 (CeCarom.); 1H NMR (400 MHz, 2.28 mmol), Pd(PPh3)4 (43.9 mg, 0.04 mmol) in 1,4-dioxane/water
DMSO-D6): d 8.93 (t, J ¼ 6.0 Hz, 1H, COeNHeCH2), 8.81 (s, 1H, H-5), (10 mL). 5c was formed as a white solid (0.259 g, 86%); mp
8.27 (s, 1H, H-3), 8.15 (s, 1H, H-300 ), 7.86 (s, 1H, H-500 ), 7.57 (d, 199.6e201.9  C; IR (KBr, nmax, cm
1): 3406 (NeHamide), 3144, 3124,
J ¼ 10.4 Hz, 2H, H-20 , H-60 ), 7.38e7.35 (m, 2H, AreH), 7.12 (t, 3091, 3061 (CeHarom.), 2861 (CeHaliph.), 1662 (C]Oamide), 1574
J ¼ 8.8 Hz, 2H, H-30 , H-50 ), 4.44 (d, J ¼ 6.4 Hz, 2H, COeNHeCH2eAr), (CeCarom.); 1H NMR (400 MHz, DMSO-D6): d 9.03 (t, J ¼ 6.4 Hz, 1H,
3.87 (s, 3H, NeCH3); 13C NMR (100 MHz, DMSO-D6): d 162.3 COeNHeCH2), 8.97e8.95 (dd, J ¼ 4.0, 1.6 Hz, 1H, H-200 ), 8.78 (s, 1H,
(COeNH), 162.1 (C-40 ), 159.8 (C-9), 142.8 (C-5), 139.7 (C-300 ), 136.0 H-5), 8.42 (s, 1H, H-3), 8.30e8.27 (dd, J ¼ 8.8, 0.8 Hz, 1H, H-800 ), 8.12
(C-10 ), 135.9 (C-2), 129.4 (C-20 ), 129.3 (C-60 ), 128.1 (C-7), 125.8 (C-6), (d, J ¼ 8.4 Hz, 1H, H-400 ), 7.86 (t, J ¼ 8.4 Hz, 1H, H-700 ), 7.70 (d,
121.9 (C-500 ), 118.8 (C-3), 117.3 (C-8), 114.9 (C-30 ), 114.7 (C-50 ), 114.6 J ¼ 8.8 Hz, 1H, H-600 ), 7.55 (d, J ¼ 10.4 Hz, 2H, H-20 , H-60 ), 7.50e7.47
(C-400 ), 41.3 (COeNHeCH2eAr), 38.7 (NeCH3); HRMS (MALDI-TOF) (dd, J ¼ 9.2, 1.6 Hz, 1H, H-300 ), 7.40e7.36 (m, 2H, AreH), 7.13 (t,
m/z calcd. for C19H16FN5O: 349.1338, found: 350.1405 [MþH]þ J ¼ 8.8 Hz, 2H, H-30 , H-50 ), 4.46 (d, J ¼ 6.4 Hz, 2H, COeNHeCH2eAr);
(100%), 372.1235 [MþNa]þ, 388.0978 [MþK]þ; Anal. calcd. for 13
C NMR (100 MHz, DMSO-D6): d 162.3 (COeNH), 162.0 (C-40 ), 159.9
C19H16FN5O: C, 65.32; H, 4.62; N, 20.05; found: C, 65.30; H, 4.611; N, (C-9), 150.6 (C-200 ), 147.9 (C-8a00 ), 143.1 (C-5), 140.1 (C-10 ), 136.0 (C-
20.09. 2), 135.4 (C-500 ), 133.5 (C-400 ), 129.4 (C-20 ), 129.3 (C-60 ), 129.1 (C-700 ),
128.9 (C-7), 127.8 (C-800 ), 127.3 (C-600 ), 126.2 (C-6), 124.3 (C-4a00 ),
4.5.1.2. N-(4-Fluorobenzyl)-6-(2-aminopyridin-3-yl)H-imidazo[1,2- 121.9 (C-300 ), 116.8 (C-3), 115.2 (C-8), 115.0 (C-30 ), 114.7 (C-50 ), 41.3
a]pyridine-2-carboxamide (5b). Prepared from 4a (300 mg, (COeNHeCH2eAr); HRMS (MALDI-TOF) m/z calcd. for C24H17FN4O:
0.76 mmol), 2-aminopyridine-3-boronic acid (157.1 mg, 1.14 mmol), 396.1386, found: 397.1459 [MþH]þ (100%), 419.1282 [MþNa]þ,
Na2CO3 (241.4 mg, 2.28 mmol), Pd(PPh3)4 (43.9 mg, 0.04 mmol) in 435.1019 [MþK]þ; Anal. calcd. for C24H17FN4O: C, 72.72; H, 4.32; N,
1,4-dioxane/water (10 mL). 5b was formed as an off white solid 14.13; found: C, 72.74; H, 4.316; N, 14.15.
(0.23 g, 84%); mp 242.0e244.6  C; IR (KBr, nmax, cm
1): 3448
(NeHamide), 3284 (NeHamine), 3175, 3138, 3065, 3038 (CeHarom.), 4.5.1.4. 3-(4-(2-((4-Fluorobenzyl)carbamoyl)H-imidazo[1,2-a]pyr-
2931 (CeHaliph.), 1651 (C]Oamide), 1606 (NeHamine), 1568 idin-6-yl)phenyl)propanoic acid (5d). Prepared from 4a (300 mg,
(CeCarom.), 1261 (CeNamine); 1H NMR (400 MHz, DMSO-D6): d 8.96 0.76 mmol), 4-(2-carboxyethyl)benzeneboronic acid (220.9 mg,
(t, J ¼ 6.4 Hz, 1H, COeNHeCH2), 8.64 (s, 1H, H-5), 8.35 (s, 1H, H-3), 1.14 mmol), Na2CO3 (241.4 mg, 2.28 mmol), Pd(PPh3)4 (43.9 mg,
7.99e7.97 (dd, J ¼ 4.8, 1.6 Hz, 1H, H-600 ), 7.63 (d, J ¼ 9.2 Hz, 1H, H-400 ), 0.04 mmol) in 1,4-dioxane/water (10 mL). 5d was formed as a white
7.42e7.33 (m, 4H, AreH), 7.12 (t, J ¼ 8.8 Hz, 2H, H-30 , H-50 ), 6.65 (t, solid (0.256 g, 81%); mp 230.1e232.7  C; IR (KBr, nmax, cm
1): 3298
J ¼ 4.8 Hz, 1H, H-500 ), 5.83 (s, 2H, AreNH2), 4.44 (d, J ¼ 6.0 Hz, 2H, (NeHamide), 3187, 3088, 3059, 3034 (CeHarom.), 2949, 2921, 2857
COeNHeCH2eAr); 13C NMR (100 MHz, DMSO-D6): d 162.3 (CeHaliph.), 2516 (OeHacid), 1703 (C]Oacid), 1646 (C]Oamide), 1580
(COeNH), 162.1 (C-40 ), 159.8 (C-9), 157.1 (AreC-200 eNH2), 147.7 (C- (CeCarom.), 1222 (CeOacid); 1H NMR (400 MHz, DMSO-D6): d 12.14
600 ), 143.1 (C-5), 139.7 (C-400 ), 137.8 (C-10 ), 136.1 (C-2), 129.3 (C-20 ), (br s, 1H, COOH), 8.96 (t, J ¼ 6.4 Hz, 1H, COeNHeCH2), 8.91 (s, 1H, H-
129.2 (C-60 ), 128.1 (C-7), 126.6 (C-6), 123.5 (C-300 ), 117.2 (C-3), 116.2 5), 8.35 (s, 1H, H-3), 7.67e7.61 (m, 4H, AreH), 7.38e7.35 (m, 4H,
(C-8), 115.0 (C-30 ), 114.7 (C-50 ), 112.7 (C-500 ), 41.3 (COeNHeCH2eAr); AreH), 7.13 (t, J ¼ 8.8 Hz, 2H, H-30 , H-50 ), 4.44 (d, J ¼ 6.4 Hz, 2H,
HRMS (MALDI-TOF) m/z calcd. for C20H16FN5O: 361.1338, found: COeNHeCH2eAr), 2.87 (t, J ¼ 7.2 Hz, 2H, CH2eCH2eAr), 2.56 (t,
362.1403 [MþH]þ (100%), 384.1236 [MþNa]þ; Anal. calcd. for J ¼ 7.6 Hz, 2H, HOOCeCH2eCH2); 13C NMR (100 MHz, DMSO-D6):
C20H16FN5O: C, 66.47; H, 4.46; N, 19.38; found: C, 66.50; H, 4.452; N, d 173.7 (COOH), 162.3 (COeNH), 162.0 (C-40 ), 159.9 (C-9), 143.1 (C-
19.33. 5), 140.8 (C-100 ), 140.0 (C-10 ), 136.0 (C-2), 133.9 (C-400 ), 129.4 (C-20 ),
129.3 (C-60 ), 129.0 (C-7), 126.4 (C-200 , C-600 ), 126.3 (C-300 , C-500 ), 125.8
4.5.1.3. N-(4-Fluorobenzyl)-6-(quinolin-5-yl)H-imidazo[1,2-a]pyri- (C-6), 124.4 (C-30 ), 117.1 (C-3), 115.0 (C-8), 114.7 (C-50 ), 41.3
dine-2-carboxamide (5c). Prepared from 4a (300 mg, 0.76 mmol), (COeNHeCH2eAr), 35.1 (CH2eCH2eCOOH), 29.9 (CH2eCH2eAr);
quinoline-5-boronic acid (197 mg, 1.14 mmol), Na2CO3 (241.4 mg, HRMS (MALDI-TOF) m/z calcd. for C24H20FN3O3: 417.1488, found:
624 G. Jose et al. / European Journal of Medicinal Chemistry 89 (2015) 616e627

418.1559 [MþH]þ (100%), 440.1385 [MþNa]þ; Anal. calcd. for 391.1172 [MþNa]þ; Anal. calcd. for C22H16N4O2: C, 71.73; H, 4.38; N,
C24H20FN3O3: C, 69.05; H, 4.83; N, 10.07; found: C, 69.09; H, 4.839; 15.21; found C, 71.76; H, 4.373; N, 15.19.
N, 10.04.
4.5.1.8. N-(Furan-2-ylmethyl)-6-(1-(2-morpholinoethyl)-1H-pyr-
4.5.1.5. N-(Furan-2-ylmethyl)-6-(1-methyl-1H-pyrazol-4-yl)H-imi- azol-4-yl)H-imidazo[1,2-a]pyridine-2-carboxamide (5h).
dazo[1,2-a]pyridine-2-carboxamide (5e). Prepared from 4b Prepared from 4b (300 mg, 0.82 mmol), 1-(2-morpholinoethyl)-1H-
(300 mg, 0.82 mmol), 1-methylpyrazole-4-boronic acid pinacol pyrazole-4-boronic acid pinacol ester (376.5 mg, 1.23 mmol),
ester (255 mg, 1.23 mmol), Na2CO3 (259.8 mg, 2.45 mmol), Na2CO3 (259.8 mg, 2.45 mmol), Pd(PPh3)4 (47.2 mg, 0.04 mmol) in
Pd(PPh3)4 (47.2 mg, 0.04 mmol) in 1,4-dioxane/water (10 mL). 5e 1,4-dioxane/water (10 mL). 5h was formed as a pale brown solid
was formed as a white solid (0.228 g, 87%); mp 188.2e190.6  C; IR (0.271 g, 79%); mp 222.4e224.7  C; IR (KBr, nmax, cm
1): 3341
(KBr, nmax, cm
1): 3286 (NeHamide), 3155, 3087 (CeHarom.), 2929 (NeHamide), 3093, 3044, (CeHarom.), 2954, 2856 (CeHaliph.), 1658
(CeHaliph.), 1655 (C]Oamide), 1577, 1565 (CeCarom.); 1H NMR (C]Oamide), 1576 (CeCarom.), 1241 (CeO); 1H NMR (400 MHz,
(400 MHz, DMSO-D6): d 8.82 (s, 1H, H-5), 8.70 (t, J ¼ 6.4 Hz, 1H, DMSO-D6): d 8.82 (s, 1H, H-5), 8.69 (t, J ¼ 6.0 Hz, 1H, COeNHeCH2),
COeNHeCH2), 8.28 (s, 1H, H-3), 8.16 (s, 1H, H-300 ), 7.86 (s, 1H, H-500 ), 8.28 (s, 1H, H-3), 8.22 (s, 1H, H-300 ), 7.88 (s, 1H, H-500 ), 7.61e7.58 (m,
7.61e7.58 (m, 2H, AreH), 7.55 (d, J ¼ 2.8 Hz, 1H, H-50 ), 6.37 (t, 2H, AreH), 7.56e7.54 (dd, J ¼ 6.4, 1.2 Hz, 1H, H-50 ), 6.37 (t, J ¼ 2.0 Hz,
J ¼ 2.0 Hz, 1H, H-40 ), 6.24 (d, J ¼ 2.8 Hz, 1H, H-30 ), 4.45 (d, J ¼ 6.4 Hz, 1H, H-40 ), 6.24 (d, J ¼ 2.8 Hz, 1H, H-30 ), 4.46 (d, J ¼ 6.4 Hz, 2H,
2H, COeNHeCH2eAr), 3.87 (s, 3H, NeCH3); 13C NMR (100 MHz, COeNHeCH2eAr), 4.25 (t, J ¼ 6.4Hz, 2H, pyrazole-NeCH2eCH2),
DMSO-D6): d 161.8 (COeNH), 152.5 (C-9), 142.7 (C-5), 141.8 (C-20 ), 3.55e3.53 (m, 4H, CH2eOeCH2), 2.73 (t, J¼ 6.4 Hz, 2H, morpholine-
139.4 (C-50 ), 135.9 (C-2), 128.1 (C-7), 126.0 (C-6), 122.0 (C-300 ), 118.9 NeCH2eCH2), 2.42e2.39 (m, 4H, CH2eNeCH2); 13C NMR (100 MHz,
(C-3), 117.8 (C-500 ), 117.3 (C-8), 114.6 (C-400 ), 110.4 (C-40 ), 106.6 (C-30 ), DMSO-D6): d 161.9 (COeNH), 152.5 (C-9), 142.8 (C-5), 141.8 (C-20 ),
38.7 (NeCH3), 35.3 (COeNHeCH2eAr); HRMS (MALDI-TOF) m/z 139.9 (C-50 ), 139.5 (C-300 ), 135.8 (C-2), 127.6 (C-7), 125.9 (C-6), 121.9
calcd. for C17H15N5O2: 321.1225, found: 322.1301 [MþH]þ (100%), (C-500 ), 118.9 (C-3), 117.4 (C-8), 114.6 (C-400 ), 110.4 (C-40 ), 106.6 (C-30 ),
344.1124 [MþNa]þ, 360.0861 [MþK]þ; Anal. calcd. for C17H15N5O2: 66.1 (CH2eOeCH2), 57.6 (CH2eNeCH2), 53.1 (pyrazole-NeCH2),
C, 63.54; H, 4.71; N, 21.79; found: C, 63.57; H, 4.703; N, 21.77. 48.8 (morpholine-NeCH2), 35.3 (COeNHeCH2eAr); HRMS
(MALDI-TOF) m/z calcd. for C22H24N6O3: 420.1909, found: 421.1985
4.5.1.6. 6-(2-Aminopyridin-3-yl)-N-(furan-2-ylmethyl)H-imidazo [MþH]þ (100%), 443.1807 [MþNa]þ; Anal. calcd. for C22H24N6O3: C,
[1,2-a]pyridine-2-carboxamide (5f). Prepared from 4b (300 mg, 62.84; H, 5.75; N, 19.99; found: C, 62.81; H, 5.742; N, 20.02.
0.82 mmol), 2-aminopyridine-3-boronic acid (269.8 mg,
1.23 mmol), Na2CO3 (259.8 mg, 2.45 mmol), Pd(PPh3)4 (47.2 mg, 4.5.1.9. N-(Furan-2-ylmethyl)-6-(4-(1-methylpiperazine-4-carbonyl)
0.04 mmol) in 1,4-dioxane/water (10 mL). 5f was formed as a white phenyl)H-imidazo[1,2-a]pyridine-2-carboxamide (5i). Prepared
crystal (0.228 g, 84%); mp 250.3e252.9  C; 1H NMR (400 MHz, from 4b (300 mg, 0.82 mmol), 4-(4-methylpiperazine-1-carbonyl)
DMSO-D6): d 8.74 (t, J ¼ 6.0 Hz, 1H, COeNHeCH2), 8.64 (s, 1H, H-5), phenyl boronic acid pinacol ester (404.8 mg, 1.23 mmol), Na2CO3
8.37 (s, 1H, H-3), 7.99e7.97 (dd, J ¼ 4.8, 1.6 Hz, 1H, H-600 ), 7.63 (d, (259.8 mg, 2.45 mmol), Pd(PPh3)4 (47.2 mg, 0.04 mmol) in 1,4-
J ¼ 9.2 Hz, 1H, H-400 ), 7.55 (d, J ¼ 2.8 Hz, 1H, H-50 ), 7.45e7.43 (dd, dioxane/water (10 mL). 5i was formed as a Pale yellow solid
J ¼ 7.2, 1.6 Hz, 1H, H-8), 7.36e7.33 (dd, J ¼ 9.6, 2.0 Hz, 1H, H-7), 6.67 (0.283 g, 78%); mp 233.0e235.3  C; 1H NMR (400 MHz, DMSO-D6):
(t, J ¼ 4.8 Hz, 1H, H-500 ), 6.38 (t, J ¼ 1.6 Hz, 1H, H-40 ), 6.24 (d, d 9.02 (s, 1H, H-5), 8.76 (t, J ¼ 5.6 Hz, 1H, COeNHeCH2), 8.39 (s, 1H,
J ¼ 3.2 Hz, 1H, H-30 ), 5.95 (s, 2H, AreNH2), 4.46 (d, J ¼ 6.0 Hz, 2H, H-3), 7.81e7.67 (m, 4H, AreH), 7.57e7.55 (m, 3H, AreH), 6.37 (t,
COeNHeCH2eAr); 13C NMR (100 MHz, DMSO-D6): d 161.9 J ¼ 2.0 Hz, 1H, H-40 ), 6.25 (d, J ¼ 2.8 Hz, 1H, H-30 ), 4.47 (d, J ¼ 5.6 Hz,
(COeNH), 156.8 (AreC-200 eNH2), 152.5 (C-9), 147.1 (C-600 ), 143.1 (C- 2H, COeNHeCH2eAr), 3.63e3.60 (m, 4H, CH2eN(CO)eCH2),
5), 141.8 (C-20 ), 139.6 (C-50 ), 138.2 (C-2), 128.1 (C-7), 126.7 (C-6), 3.01e2.70 (m, 4H, CH2eN(CH3)eCH2), 2.56 (s, 3H, NeCH3); 13C
123.3 (C-400 ), 117.2 (C-3), 116.5 (C-8), 115.1 (C-300 ), 112.7 (C-500 ), 110.4 NMR (100 MHz, DMSO-D6): d 168.6 (piperazine-NeCOeAr), 161.8
(C-40 ), 106.6 (C-30 ), 35.4 (COeNHeCH2eAr); HRMS (MALDI-TOF) m/ (COeNH), 152.5(C-9), 143.2 (C-5), 141.8 (C-20 ), 139.9 (C-50 ), 137.6 (C-
z calcd. for C18H15N5O2: 333.1225, found: 334.1301 [MþH]þ (100%), 100 ), 134.6 (C-2), 127.9 (C-7), 126.6 (C-400 ), 126.2 (C-6), 125.2 (C-300 ,C-
356.1121 [MþNa]þ, 372.0860 [MþK]þ; Anal. calcd. for C18H15N5O2: 500 ), 125.1 (C-200 ,C-600 ), 117.4 (C-3), 115.3 (C-8), 110.4 (C-40 ), 106.7 (C-
C, 64.86; H, 4.54; N, 21.01; found C, 64.88; H, 4.549; N, 21.03. 30 ), 53.1 (CH2eN(CH3)eCH2), 45.5 (CH2eN(CO)eCH2), 43.7
(NeCH3), 35.3 (COeNHeCH2eAr); HRMS (MALDI-TOF) m/z calcd.
4.5.1.7. N-(Furan-2-ylmethyl)-6-(quinolin-5-yl)H-imidazo[1,2-a]pyr- for C25H25N5O3: 443.1957, found: 444.2031 [MþH]þ (100%),
idine-2-carboxamide (5g). Prepared from 4b (300 mg, 0.82 mmol), 466.1853 [MþNa]þ; Anal. calcd. for C25H25N5O3: C, 67.70; H, 5.68; N,
quinoline-5-boronic acid (212 mg, 1.23 mmol), Na2CO3 (259.8 mg, 15.79; found: C, 67.68; H, 5.689; N, 15.80.
2.45 mmol), Pd(PPh3)4 (47.2 mg, 0.04 mmol) in 1,4-dioxane/water
(10 mL). 5g was formed as an off white solid (0.258 g, 86%); mp 4.5.1.10. 6-(4-Chlorophenyl)-N-(pyridin-4-ylmethyl)H-imidazo[1,2-
201.9e204.1  C; IR (KBr, nmax, cm
1): 3401 (NeHamide), 3124 a]pyridine-2-carboxamide (5j). Prepared from 4c (300 mg,
(CeHarom.), 2862 (CeHaliph.), 1665 (C]Oamide), 1572 (CeCarom.); 1H 0.79 mmol), 4-chlorophenylboronic acid (186.1 mg, 1.19 mmol),
NMR (400 MHz, DMSO-D6): d 8.97e8.95 (dd, J ¼ 4.0, 1.6 Hz, 1H, H- Na2CO3 (252.2 mg, 2.38 mmol), Pd(PPh3)4 (45.8 mg, 0.04 mmol) in
200 ), 8.85 (t, J ¼ 6.0 Hz, 1H, COeNHeCH2), 8.80 (s, 1H, H-5), 8.44 (s, 1,4-dioxane/water (10 mL). 5j was formed as a white solid (0.25 g,
1H, H-3), 8.29 (d, J ¼ 8.4 Hz, 1H, H-800 ), 8.12 (d, J ¼ 8.8 Hz, 1H, H-400 ), 87%); mp 166.8e169.0  C; IR (KBr, nmax, cm
1): 3409 (NeHamide),
7.86 (t, J ¼ 7.2 Hz, 1H, H-700 ), 7.74e7.67 (m, 2H, AreH), 7.57e7.47 (m, 3199, 3136, 3071, 3033 (CeHarom.), 2919 (CeHaliph.), 1645 (C]Oamide),
3H, AreH), 6.39 (t, J ¼ 2.0 Hz, 1H, H-40 ), 6.26 (d, J ¼ 2.4 Hz, 1H, H-30 ), 1602, 1566 (CeCarom.); 1H NMR (400 MHz, DMSO-D6): d 9.11 (t,
4.49 (d, J ¼ 6.0 Hz, 2H, COeNHeCH2eAr); 13C NMR (100 MHz, J ¼ 6.4 Hz, 1H, COeNHeCH2), 8.97 (s, 1H, H-5), 8.49 (d, J ¼ 5.2 Hz, 2H,
DMSO-D6): d 161.8 (COeNH), 152.5 (C-9), 150.6 (C-200 ), 149.7 (C- H-20 , H-60 ), 8.37 (s, 1H, H-3), 7.74 (d, J ¼ 8.4 Hz, 2H, H-200 , H-600 ), 7.68
8a00 ), 147.9 (C-500 ), 143.1 (C-5), 141.8 (C-20 ), 139.9 (C-50 ), 136.7 (C-400 ), (m, 2H, AreH), 7.56 (d, J ¼ 8.4 Hz, 2H, H-300 , H-500 ), 7.30 (d, J ¼ 5.6 Hz,
135.4 (C-2), 133.5 (C-700 ), 130.4 (C-800 ), 129.4 (C-600 ), 129.1 (C-4a00 ), 2H, H-30 , H-50 ), 4.49 (d, J ¼ 6.0 Hz, 2H, COeNHeCH2eAr); 13C NMR
127.8 (C-7), 126.2 (C-6), 124.3(C-300 ), 121.9 (C-3), 116.8 (C-8), 110.4 (100 MHz, DMSO-D6): d 162.3 (COeNH), 149.4 (C-9), 148.7 (C-20 , C-
(C-30 ), 106.6 (C-40 ), 35.4 (COeNHeCH2eAr); HRMS (MALDI-TOF) m/ 60 ), 143.2 (C-5), 139.9 (C-40 ), 135.1 (C-2), 132.8 (C-100 ), 129.1 (C-400 ),
z calcd. for C22H16N4O2: 368.1273, found: 369.1349 [MþH]þ (100%), 128.3 (C-300 , C-500 ), 126.2 (C-200 , C-600 ), 125.0 (C-7), 124.7 (C-6), 122.2
 G. Jose et al. / European Journal of Medicinal Chemistry 89 (2015) 616e627 625

(C-30 , C-50 ), 117.3 (C-3), 115.3 (C-8), 41.1 (COeNHeCH2eAr); HRMS (CH2eNeCH2), 53.0 (pyrazole-NeCH2), 48.8 (morpholine-NeCH2),
(MALDI-TOF) m/z calcd. for C20H15ClN4O: 362.0934, found: 363.1010 24.9 (C-10 ), 5.7 (C-20 ), 5.5 (C-30 ); HRMS (MALDI-TOF) m/z calcd. for
[MþH]þ (100%), 385.0833 [MþNa]þ; Anal. calcd. for C20H15ClN4O: C, C20H24N6O2: 380.1961, found: 381.2035 [MþH]þ (100%), 403.1859
66.21; H, 4.17; N, 15.44; found: C, 66.19; H, 4.179; N, 15.47. [MþNa]þ; Anal. calcd. for C20H24N6O2: C, 63.14; H, 6.36; N, 22.09;
found: C, 63.19; H, 6.370; N, 22.10.
4.5.1.11. 6-(2-Methoxyphenyl)-N-(pyridin-4-ylmethyl)H-imidazo
[1,2-a]pyridine-2-carboxamide (5k). Prepared from 4c (300 mg, 4.5.1.14. N-(4-(Trifluoromethyl)benzyl)-6-(quinolin-5-yl)H-imidazo
0.79 mmol), 2-methoxyphenylboronic acid (180.8 mg, 1.19 mmol), [1,2-a]pyridine-2-carboxamide (5n). Prepared from 4f (300 mg,
Na2CO3 (252.2 mg, 2.38 mmol), Pd(PPh3)4 (45.8 mg, 0.04 mmol) in 0.67 mmol), quinoline-5-boronic acid (175 mg, 1.01 mmol), Na2CO3
1,4-dioxane/water (10 mL). 5k was formed as a white solid (0.244 g, (214.3 mg, 2.02 mmol), Pd(PPh3)4 (39 mg, 0.03 mmol) in 1,4-
86%); mp 144.5e146.9  C; IR (KBr, nmax, cm
1): 3359 (NeHamide), dioxane/water (10 mL). 5n was formed as a white solid (0.26 g,
3139, 3091, 3029, 3001 (CeHarom.), 2936, 2837 (CeHaliph.), 1664 (C] 86%); mp 177.1e179.6  C; IR (KBr, nmax, cm
1): 3404 (NeHamide),
Oamide), 1600, 1570 (CeCarom.); 1H NMR (400 MHz, DMSO-D6): d 9.10 3144 (CeHarom.), 1666 (C]Oamide), 1570 (CeCarom.), 1328 (CeF); 1H
(t, J ¼ 6.4 Hz, 1H, COeNHeCH2), 8.71 (s, 1H, H-5), 8.49 (d, J ¼ 6.0 Hz, NMR (400 MHz, DMSO-D6): d 9.16 (t, J ¼ 6.4 Hz, 1H, COeNHeCH2),
2H, H-20 , H-60 ), 8.41 (s, 1H, H-3), 7.61 (d, J ¼ 9.2 Hz, 1H, H-600 ), 8.97e8.95 (dd, J ¼ 4.0, 1.6 Hz, 1H, H-200 ), 8.79 (s, 1H, H-5), 8.43 (s, 1H,
7.51e7.48 (dd, J ¼ 9.6, 1.6 Hz, 1H, H-400 ), 7.42e7.38 (m, 2H, AreH), H-3), 8.29 (d, J ¼ 8.4 Hz, 1H, H-800 ), 8.12 (d, J ¼ 8.4 Hz, 1H, H-400 ), 7.86
7.30 (d, J ¼ 5.6 Hz, 2H, H-30 , H-50 ), 7.15 (t, J ¼ 4.4 Hz, 1H, H-500 ), 7.06 (t, J ¼ 8.4 Hz, 1H, H-700 ), 7.67 (d, J ¼ 8.0 Hz, 2H, H-30 , H-50 ), 7.53 (d,
(d, J ¼ 6.8 Hz, 1H, H-300 ), 4.49 (d, J ¼ 6.4 Hz, 2H, COeNHeCH2eAr), J ¼ 8.0 Hz, 2H, H-20 , H-60 ), 7.40e7.36 (m, 2H, AreH), 7.16e7.11 (m,
3.80 (s, 3H, AreOeCH3); 13C NMR (100 MHz, DMSO-D6): d 162.5 2H, AreH), 4.56 (d, J ¼ 6.0 Hz, 2H, COeNHeCH2eAr); 13C NMR
(COeNH), 156.4 (C-200 eOeCH3), 149.4 (C-9), 148.7 (C-20 , C-60 ), 142.9 (100 MHz, DMSO-D6): d 162.3 (COeNH), 150.7 (C-200 ), 147.9 (C-8a00 ),
(C-5), 139.5 (C-40 ), 135.2 (C-2), 129.6 (C-400 ), 129.0 (C-600 ), 126.4 (C- 144.7 (C-9), 143.2 (C-5), 139.9 (C-100 ), 135.4 (C-2), 133.5 (C-10 ), 129.4
100 ), 125.3 (C-7), 123.8 (C-6), 122.2 (C-30 , C-50 ), 120.8 (C-500 ), 116.0 (C- (C-400 ), 129.1 (C-700 ), 129.0 (C-800 ), 127.9 (C-7), 127.8 (C-40 ), 127.5 (C-
3), 115.1 (C-8), 111.8 (C-300 ), 55.6 (AreOeCH3), 41.2 20 , C-60 ), 127.3 (C-600 ), 126.2 (C-6), 125.1 (C-30 , C-50 ), 125.0 (AreCF3),
(COeNHeCH2eAr); HRMS (MALDI-TOF) m/z calcd. for C21H18N4O2: 124.3 (C-4a00 ), 121.9 (C-300 ), 116.8 (C-3), 115.2 (C-8), 41.7
358.1429, found: 359.1505 [MþH]þ (100%), 381.1326 [MþNa]þ; (COeNHeCH2eAr); HRMS (MALDI-TOF) m/z calcd. for
Anal. calcd. for C21H18N4O2: C, 70.38; H, 5.06; N, 15.63; found: C, C25H17F3N4O: 446.1354, found: 447.1433 [MþH]þ (100%), 469.1251
70.40; H, 5.055; N, 15.61. [MþNa]þ, 485.0989 [MþK]þ; Anal. calcd. for C25H17F3N4O: C, 67.26;
H, 3.84; N, 12.55; found: C, 67.28; H, 3.833; N, 12.52.
4.5.1.12. N-(Benzo[d][1,3]dioxol-5-ylmethyl)-6-(1-methyl-1H-pyr-
azol-4-yl)H-imidazo[1,2-a]pyridine-2-carboxamide (5l). 4.5.1.15. N-(4-(Trifluoromethyl)benzyl)-6-(1-(2-morpholinoethyl)-
Prepared from 4d (300 mg, 0.71 mmol), 1-methylpyrazole-4- 1H-pyrazol-4-yl)H-imidazo[1,2-a]pyridine-2-carboxamide (5o).
boronic acid pinacol ester (222.3 mg, 1.07 mmol), Na2CO3 Prepared from 4f (300 mg, 0.67 mmol), 1-(2-morpholinoethyl)-1H-
(226.5 mg, 2.14 mmol), Pd(PPh3)4 (41.2 mg, 0.04 mmol) in 1,4- pyrazole-4-boronic acid pinacol ester (310.5 mg, 1.01 mmol),
dioxane/water (10 mL). 5l was formed as a white solid (0.225 g, Na2CO3 (214.3 mg, 2.02 mmol), Pd(PPh3)4 (39 mg, 0.03 mmol) in
84%); mp 251.0e253.4  C; IR (KBr, nmax, cm
1): 3343 (NeHamide), 1,4-dioxane/water (10 mL). 5o was formed as a pale yellow solid
3148, 3087 (CeHarom.), 2940, 2921 (CeHaliph.), 1654 (C]Oamide), (0.265 g, 79%); mp 199.9e202.6  C; IR (KBr, nmax, cm
1): 3344
1579 (CeCarom.), 1243 (CeOether); 1H NMR (400 MHz, DMSO-D6): (NeHamide), 3140 (CeHarom.), 2994, 2856 (CeHaliph.), 1645 (C]
d 8.83 ((t, J ¼ 6.4 Hz, 1H, COeNHeCH2), 8.81 (s, 1H, H-5), 8.26 (s, 1H, Oamide), 1578 (CeCarom.), 1330 (CeF); 1H NMR (400 MHz, DMSO-
H-3), 8.15 (s, 1H, H-300 ), 7.86 (s, 1H, H-500 ), 7.59e7.54 (m, 2H, AreH), D6): d 9.07 (t, J ¼ 6.4 Hz, 1H, COeNHeCH2), 8.82 (s, 1H, H-5), 8.29 (s,
6.91 (s, 1H, H-40 ), 6.84e6.78 (m, 2H, AreH), 5.95 (s, 2H, OeCH2eO), 1H, H-3), 8.22 (s, 1H, H-300 ), 7.88 (s, 1H, H-500 ), 7.67 (d, J ¼ 8.0 Hz, 2H,
4.36 (d, J ¼ 6.4 Hz, 2H, COeNHeCH2eAr), 3.87 (s, 3H, NeCH3); 13C H-30 , H-50 ), 7.60e7.59 (m, 2H, AreH), 7.53 (d, J ¼ 8.0 Hz, 2H, H-20 , H-
NMR (100 MHz, DMSO-D6): d 161.9 (COeNH), 147.1 (C-9), 145.9 (H- 60 ), 4.55 (d, J ¼ 6.4 Hz, 2H, COeNHeCH2eAr), 4.25 (t, J ¼ 6.4 Hz, 2H,
3a0 ), 142.8 (C-5), 139.8 (C-300 ), 135.9 (C-2), 133.7 (H-7a0 ), 128.1 (C-7), pyrazole-NeCH2eCH2), 3.54 (t, J ¼ 4.8 Hz, 4H, CH2eOeCH2), 2.73 (t,
125.8 (C-6), 121.9 (C-500 ), 120.6 (C-50 ), 118.8 (C-3), 117.8 (C-60 ), 117.3 J ¼ 6.4 Hz, 2H, morpholine-NeCH2eCH2), 2.41 (t, J ¼ 4.4 Hz, 4H,
(C-8), 114.5 (C-400 ), 108.1 (C-70 ), 107.9 (C-40 ), 100.7 (OeCH2eO), 41.7 CH2eNeCH2); 13C NMR (100 MHz, DMSO-D6): d 162.3 (COeNH),
(COeNHeCH2eAr), 38.7 (NeCH3); HRMS (MALDI-TOF) m/z calcd. 144.7 (C-9), 142.8 (C-5), 139.6 (C-300 ), 135.8 (C-2), 133.5 (C-10 ), 127.9
for C20H17N5O3: 375.1331, found: 376.1402 [MþH]þ (100%), (C-7), 127.6 (C-40 ), 127.5 (C-20 , C-60 ), 125.9 (C-6), 125.1 (C-30 , C-50 ),
398.1228 [MþNa]þ; Anal. calcd. for C20H17N5O3: C, 63.99; H, 4.56; 125.0 (AreCF3), 121.9 (C-500 ), 118.9 (C-3), 117.3 (C-8), 114.7 (C-400 ),
N, 18.66; found: C, 64.01; H, 4.553; N, 18.67. 66.1 (CH2eOeCH2), 57.6 (CH2eNeCH2), 53.1 (pyrazole-NeCH2),
48.8 (morpholine-NeCH2), 41.6 (COeNHeCH2eAr); HRMS (MALDI-
4.5.1.13. N-Cyclopropyl-6-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl) TOF) m/z calcd. for C25H25F3N6O2: 498.1991, found: 499.2065
H-imidazo[1,2-a]pyridine-2-carboxamide (5m). Prepared from 4e [MþH]þ (100%), 521.1889 [MþNa]þ, 537.1622 [MþK]þ; Anal. calcd.
(300 mg, 0.92 mmol), 1-(2-morpholinoethyl)-1H-pyrazole-4- for C25H25F3N6O2: C, 60.23; H, 5.05; N, 16.86; found: C, 60.20; H,
boronic acid pinacol ester (422.6 mg, 1.38 mmol), Na2CO3 (291.6 mg, 5.044; N, 16.89.
2.75 mmol), Pd(PPh3)4 (53 mg, 0.05 mmol) in 1,4-dioxane/water
(10 mL). 5m was formed as a pale brown solid (0.269 g, 77%); mp 4.5.1.16. 6-(1-Methyl-1H-pyrazol-4-yl)-N-(tetrahydro-2H-pyran-4-
135.1e137.6  C; 1H NMR (400 MHz, DMSO-D6): d 8.81 (s, 1H, H-5), yl)H-imidazo[1,2-a]pyridine-2-carboxamide (5p). Prepared from 4g
8.30 (d, J ¼ 4.8 Hz, 1H, COeNHeCH), 8.25 (s, 1H, H-3), 8.21 (s, 1H, H- (300 mg, 0.81 mmol), 1-methyl-1H-pyrazole-4-boronic acid pina-
300 ), 7.87 (s, 1H, H-500 ), 7.59e7.54 (m, 2H, AreH), 4.25 (t, J ¼ 6.4 Hz, 2H, col ester (252.3 mg, 1.21 mmol), Na2CO3 (257 mg, 2.42 mmol),
pyrazole-NeCH2eCH2), 3.55e3.51 (m, 4H, CH2eOeCH2), 2.89e2.85 Pd(PPh3)4 (46.7 mg, 0.04 mmol) in 1,4-dioxane/water (10 mL). 5p
(m, 1H, H-10 ), 2.73 (t, J ¼ 6.4 Hz, 2H, morpholine-NeCH2eCH2), was formed as an off white solid (0.216 g, 82%); mp 125.0e127.4  C;
2.42e2.37 (m, 4H, CH2eNeCH2), 0.67e0.64 (m, 4H, H-20 , H-30 ); 13C IR (KBr, nmax, cm
1): 3335 (NeHamide), 3153, 3124, 3043 (CeHarom.),
NMR (100 MHz, DMSO-D6): d 163.2 (COeNH), 144.3 (C-9), 142.7 (C- 2952, 2929, 2839 (CeHaliph.), 1640 (C]Oamide), 1558 (CeCarom.); 1H
5), 139.8 (C-300 ), 138.3 (C-2), 135.8 (C-7), 127.6 (C-6), 125.8 (C-3), 121.9 NMR (400 MHz, DMSO-D6): d 8.80 (s, 1H, H-5), 8.25 (s, 1H, H-3), 8.17
(C-500 ), 117.2 (C-8), 114.3 (C-400 ), 66.1 (CH2eOeCH2), 57.6 (d, J ¼ 8.4 Hz, 1H, COeNHeCH), 8.15 (s, 1H, H-300 ), 7.86 (s, 1H, H-500 ),
626 G. Jose et al. / European Journal of Medicinal Chemistry 89 (2015) 616e627

7.60e7.55 (m, 2H, AreH), 4.04e3.97 (m, 1H, H-40 ), 3.87 (s, 3H, control was also prepared on each plate. Sterile water was added to
NeCH3), 3.85 (d, J ¼ 8.8 Hz, 2H, H-2a0 , H-6a0 ), 3.41e3.34 (dt, J ¼ 11.2, all perimeter wells to avoid evaporation during the incubation. The
3.2 Hz, 2H, H-2b0 , H-6b0 ), 1.71e1.65 (m, 4H, H-30 , H-50 ); 13C NMR plate was covered, sealed in plastic bags and incubated at 37  C in
(100 MHz, DMSO-D6): d 161.2 (COeNH), 147.9 (C-9), 142.7 (C-5), normal atmosphere. After 7 days incubation, 30 mL of alamar blue
139.8 (C-300 ), 135.9 (C-2), 128.1 (C-7), 125.9 (C-6), 121.9 (C-500 ), 118.8 solution was added to each well, and the plate was re-incubated
(C-3), 117.2 (C-8), 114.5 (C-400 ), 66.1 (C-20 , C-60 ), 44.9 (C-40 ), 38.7 overnight. A change in colour from blue (oxidised state) to pink
(NeCH3), 32.3 (C-30 , C-50 ); HRMS (MALDI-TOF) m/z calcd. for (reduced) indicated the growth of bacteria, and the MIC was defined
C17H19N5O2: 325.1538, found: 326.1611 [MþH]þ (100%), 348.4138 as the lowest concentration of drug that prevented this change in
[MþNa]þ; Anal. calcd. for C17H19N5O2: C, 62.75; H, 5.89; N, 21.52; colour. Isoniazid and Rifampicin were used as standards.
found: C, 62.74; H, 5.882; N, 21.50.
4.8. Cytotoxicity
4.5.1.17. 6-(Quinolin-5-yl)-N-(tetrahydro-2H-pyran-4-yl)H-imidazo
[1,2-a]pyridine-2-carboxamide (5q). Prepared from 4g (300 mg, All the new compounds synthesized were further examined for
0.81 mmol), quinoline-5-boronic acid (209.7 mg, 1.21 mmol), cytotoxicity against Human Embryonic Kidney Cell-line 293T (HEK-
Na2CO3 (257 mg, 2.42 mmol), Pd(PPh3)4 (46.7 mg, 0.04 mmol) in 293T) cells at the concentration of 50 mg/mL. After 72 h of expo-
1,4-dioxane/water (10 mL). 5q was formed as an off white solid sure, viability was assessed on the basis of cellular conversion of
(0.253 g, 84%); mp 143.8e146.0  C; IR (KBr, nmax, cm
1): 3337 (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
(NeHamide), 3099, 3066, 3034 (CeHarom.), 2936, 2848 (CeHaliph.), (MTT) into a formazan product using the Promega Cell Titer 96 non-
1644 (C]Oamide), 1563 (CeCarom.); 1H NMR (400 MHz, DMSO-D6): radioactive cell proliferation assay.
d 8.96e8.95 (dd, J ¼ 4.4, 1.6 Hz, 1H, H-200 ), 8.77 (s, 1H, H-5), 8.41 (s,
1H, H-3), 8.29 (d, J ¼ 8.8 Hz, 1H, H-800 ), 8.17 (d, J ¼ 8.4 Hz, 1H,
4.9. Molecular docking study procedure
COeNHeCH), 8.12 (d, J ¼ 8.4 Hz, 1H, H-400 ), 7.86 (t, J ¼ 8.4 Hz, 1H, H-
700 ), 7.73e7.66 (m, 2H, AreH), 7.55 (d, J ¼ 8.8 Hz, 1H, H-7), 7.49e7.47
The three dimensional structure of target protein InhA, the
(dd, J ¼ 9.2, 1.6 Hz, 1H, H-300 ), 4.08e3.98 (m, 1H, H-40 ), 3.87 (d,
enoyl acyl carrier protein reductase (ENR) from M. tuberculosis, is
J ¼ 11.2 Hz, 2H, H-2a0 , H-6a0 ), 3.41e3.35 (dt, J ¼ 11.2, 4.0 Hz, 2H, H-
one of the key enzymes involved in mycobacterial fatty acid elon-
2b0 , H-6b0 ), 1.74e1.66 (m, 4H, H-30 , H-50 ); 13C NMR (100 MHz,
gation cycle, which has been validated as an effective antimicrobial
DMSO-D6): d 161.2 (COeNH), 150.6 (C-200 ), 147.9 (C-9), 143.0 (C-5),
target, having keyword 2X22 was downloaded from PDB (www.
140.3 (C-8a00 ), 135.4 (C-2), 133.4 (C-500 ), 129.4 (C-400 ), 129.0 (C-700 ),
rcsb.org/pdb) structural database. This file was then opened in
128.9 (C-800 ), 127.8 (C-7), 127.2 (C-600 ), 126.1 (C-6), 124.2 (C-4a00 ),
SPDB viewer edited by removing the heteroatoms, adding C ter-
121.9 (C-300 ), 116.7 (C-3), 115.1 (C-8), 66.2 (C-20 , C-60 ), 45.1 (C-40 ),
minal oxygen. The active pockets on target protein molecule were
32.4 (C-30 , C-50 ); HRMS (MALDI-TOF) m/z calcd. for C22H20N4O2:
found out using CASTp server [43]. The ligands were drawn using
372.1586, found: 373.1661 [MþH]þ (100%), 395.1482 [MþNa]þ
ChemDraw Ultra 6.0 and assigned with proper 2D orientation
395.1484; Anal. calcd. for C22H20N4O2: C, 70.95; H, 5.41; N, 15.04;
(ChemOffice package). 3D coordinates were prepared using
found: C, 70.93; H, 5.403; N, 15.03.
PRODRG server [44]. Autodock V3.0 was used to perform Auto-
mated Molecular Docking in AMD Athlon (TM)2x2 215 at 2.70 GHz,
4.6. X-ray crystallographic analysis
with 1.75 GB of RAM. AutoDock 3.0 was compiled and run under
Microsoft Windows XP service pack 3. For docking, grid map re-
The X-ray crystallographic analysis of 5f was carried out on a
quires in AutoDock, the size of the grid box was set at 114, 116 and
white crystal, with dimensions 0.21 mm  0.18 mm  0.26 mm,
100 Å (R, G, and B), and grid center
12.755,
22.815, 32.099 for x,
grown from the slow evaporation of a dichloromethane and
y, and z-coordinates. All torsions were allowed to rotate during
methanol solvent mixture at room temperature. A suitable crystal
docking. The Lamarckian genetic algorithm and the pseudo-Solis
was selected and mounted on a Bruker APEX-II CCD diffractometer.
and Wets methods were applied for minimization, using default
The crystal was kept at 296.15 K during data collection. Using Olex2
parameters [45]. The newly synthesized compounds were taken as
[39], the structure was solved the Superflip [40] structure solution
ligands and docked against target molecule InhA.
program using Charge Flipping and refined with the ShelXL [41]
refinement package using Least Squares minimization. CCDC
Acknowledgements
1007948 contains the supplementary crystallographic data for 5f.
These data can be obtained free of charge via http://www.ccdc.cam.
The authors are thankful to the authorities of Jain University for
ac.uk/conts/retrieving.html, or from the Cambridge Crystallo-
encouragement and financial support for this work. GJ acknowl-
graphic Data Centre, 12, Union Road, Cambridge CB2 1EZ, UK; fax:
edges the NMR Research Centre, Indian Institute of Science, Ban-
(þ44) 1223-336-033; or email: [email protected].
galore, for providing spectral data.
4.7. In vitro Mtb MABA assay procedure
Appendix A. Supplementary data
The anti-mycobacterial activities of compounds 5aeq have been
assessed against M. tuberculosis ATCC 27294 using the microplate Supplementary data associated with this article can be found in
alamar blue assay (MABA) [42]. Briefly, the inoculum was prepared the online version, at http://dx.doi.org/10.1016/j.ejmech.2014.10.
from fresh LJ medium re-suspended in 7H9-S medium (7H9 broth, 079. These data include MOL files and InChiKeys of the most
0.1% casitone, 0.5% glycerol, supplemented oleic acid, albumin, important compounds described in this article.
dextrose, and catalase [OADC]), adjusted to a McFarland tube No. 1,
and diluted 1:20; 100 mL was used as inoculum. Each drug stock References
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