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Inhibition

Leishmaniadonovani
Anti-promastigote activity. L. donovani promastigote inhibition potency of these reported β-carboline derivatives was determined by Alamar blue assay method. Among these analogues, compound 7d exhibited potent inhibition (EC50 0.91 µM) of L. donovani promastigotes than standard drugs pentamidine and miltefosine (EC50 6.4 and 3.12 µM). Anti-promastigote activity of analogues 7h, 7n and 7g (EC50 4.0, 4.57, and 5.02 µM respectively) is equipotent as that of standard drugs. Compounds 7f, 7p, 7c, 7a, 7k, 7j, 7b, 7i, and 7odisplayedsignificant inhibition (EC50 8.42, 8.5, 9.45, 11.9, 12.4, 13.1, 14.2, 15.2 and 19.5 µM respectively) of promastigote and inhibition potency is comparable with standard drugs. Compound 7e showed moderate (EC50 28.5 µM) anti-promastigote activity and 7l and 7m displayed weak inhibition (EC50 85.9 and 117.5 µM) of L. donovani promastigotes.

 

Axenic amastigote screening.

Compounds exhibited significant anti-promastigote activity (EC50 < 20 µM), were further screened against axenic amastigote (human life cycle parasite form) forms. Among these derivatives, Analogues 7d, 7h, 7n and 7g showed potent inhibition (EC50 0.9, 2.2 3.5 and 3.8 µM respectively) of axenic amastigotes. Especially, compound 7d exhibited two to three times more potent anti-amastigote activity than pentamidine and miltefosine (EC50 1.6 and 2.8 µM). Compounds 7c, 7b, 7f, 7j, 7k and 7a exhibited significant inhibition (EC50 5.3, 5.8, 6.4, 7.5, 9.3 and 10.0 µM respectively) of L. donovani axenic amastigotes. Compounds 7p, 7o and 7i displayed moderate anti-amastigote activity (EC50 12.3, 13.9 and 19.4 µM respectively). Axenic amastigote assay results revealed that these β-carboline derivatives exhibited potent inhibition of axenic amastigotes of L. donovani.

 

Intracellular amastigote screening.

Although, axenic amastigotes assay most commonly used to screen new leishmanial agents are different from intracellular amastigotes in terms of protein expression and drug susceptibility. Hence in our present study, compounds displayed potent inhibition of promastigote and axenic amastigotes were further evaluated against intracellular amastigotes to ensure their activity against clinical relevant forms of parasite.

 

Intracellular amastigote assay results suggested that, most of these analogues displayed potent inhibition of L. donovanimacrophagic amastigotesand the potency was comparable with miltefosine and many folds higher than pentamidine (Table 2). Especially, compound 7d ((4-(4-methoxyphenyl)piperazin-1-yl)(9-methyl-1-phenyl-9H-pyrido [3,4-b]indol-3-yl)methan-one) exhibited five times potent inhibition (EC50 1.3 µM) of intracellular amastigotes compared to miltefosine (EC50 6.4 µM). Compounds 7n and 7g were displayed potent antiamastigote activity (EC50 5.6 and 6.3 µM) than standard drugs
miltefosine and pentamidine with better selectivity index. Antiamastigote potency of 7b, 7c, 7h, 7o and 7p (EC50 7.2, 7.8, 7.9, 7.9 and 9.4 µM) is comparable with miltefosine and better than pentamidine. Compounds 7k, 7f, 7a, 7j and 7i exhibited good antiamastigote activity (EC50 10.1, 11.8, 13.5, 15.4 and 22.0 µM) with better selectivity.

 

Structure Activity Relationship (SAR).

Structure activity relationship study suggests that, substitution or replacement of phenyl ring has significant effect on anti-leishmanial activity as well as cytotoxicity of these analogues. Among these analogues, un-substituted phenyl derivative displayed significant inhibition of promastigotes, axenic amastigote and intracellular amastigotes of L. donovani. Ortho-para directing methyl group substitution on ortho and para position enhanced the potency against axenic amastigotes and intracellular amastigotes of L. donovani significantly, while on para substitution anti-promastigote activity altered marginally. Methoxy, chloro group substitution had strong impact on anti-leishmanial activity of these analogues, position of substitution is vital in their inhibition activity. Ortho substitution of
methoxy group showed excellent increase in inhibition potency against promastigotes and both amastigote forms of L. donovani. While methoxy substitution on ortho position slightly increased the anti-leishmanial activity, meta substitution resulted in drastic decline in activity. Chloro substitution on para and meta position has significantly increased the inhibition potency against tested forms, while on ortho substitution has marginally affected the activity. Meta directing nitro and fluoro substitution on para position haven’t showed any considerable effect on inhibition potency of these analogues.

 

However, fluoro substitution on ortho position and chloro di-substitution (ortho and meta) significantly decreased the anti-leishmanial potency. Replacement of phenyl ring with benzyl ring has significantly enhanced the activity. Even though pyridyl replacement resulted in increased anti-leishmanial activity, it also enhanced the cytotoxicity of these analogues. SAR studies suggested that, ortho-para directing methoxy group on para position, chloro group on para and meta position and benzyl replacement was recommended for anti-leishmanial activity of these analogues against L.
donovani.

 

Through the present study, we have identified new lead molecules with potent anti-leishmaninal activity against L. infantumand L. donovani. Structure activity relationship studies would be utilised to optimize the lead molecules to generate clinically effective against both drug sensitive and resistance strains of L. infantumand L. donovani.

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MF:C16H14N4 MW:262.3092

CAS No. 101987-76-2

3-Quinolinecarboxylicacid,8-chloro-1-cyclopropyl-6-fluoro-1,4-dihydro-7-(3-methyl-1-piperazinyl)-4-oxo-

Catalog No.:AG00065X MDL No.:

MF:C18H19ClFN3O3 MW:379.8132

CAS No. 101987-86-4

Benzenepropanoic acid, 3-chloro-2,4,5-trifluoro-β-oxo-, ethyl ester

Catalog No.:AG00065W MDL No.:MFCD09037898

MF:C11H8ClF3O3 MW:280.6276

CAS No. 101987-89-7

3-Quinolinecarboxylic acid, 8-chloro-1-cyclopropyl-6,7-difluoro-1,4-dihydro-4-oxo-

Catalog No.:AG00065V MDL No.:MFCD08706388

MF:C13H8ClF2NO3 MW:299.6573

CAS No. 101988-70-9

1,3,5-Triazine-2,4-diamine, N2-methyl-6-(2,2,2-trifluoroethoxy)-

Catalog No.:AG00065U MDL No.:MFCD00452913

MF:C6H8F3N5O MW:223.1558

CAS No. 10199-00-5

2,2'-BIPYRAZINE

Catalog No.:AG000666 MDL No.:MFCD00010288

MF:C8H6N4 MW:158.1600

CAS No. 10199-50-5

1H-Pyrazol-5-amine, 1-methyl-3-phenyl-

Catalog No.:AG000665 MDL No.:MFCD00067874

MF:C10H11N3 MW:173.2144

CAS No. 10199-51-6

1H-Pyrazole-3-carboxylic acid, 1-methyl-5-phenyl-, ethyl ester

Catalog No.:AG000664 MDL No.:MFCD08060533

MF:C13H14N2O2 MW:230.2625

CAS No. 10199-53-8

1H-Pyrazole-3-carboxylic acid, 1-methyl-5-phenyl-

Catalog No.:AG000671 MDL No.:MFCD08271933

MF:C11H10N2O2 MW:202.2093

CAS No. 10199-57-2

1H-Pyrazole-3-carboxylic acid, 5-methyl-1-phenyl-

Catalog No.:AG000670 MDL No.:MFCD00124910

MF:C11H10N2O2 MW:202.2093

CAS No. 10199-59-4

1H-Pyrazole-1-carboxylic acid, ethyl ester

Catalog No.:AG00066Z MDL No.:

MF:C6H8N2O2 MW:140.1399

CAS No. 10199-61-8

1H-Pyrazole-1-acetic acid, ethyl ester

Catalog No.:AG00066Y MDL No.:MFCD00159620

MF:C7H10N2O2 MW:154.1665

CAS No. 10199-63-0

Ethanone, 1-(3,5-dimethyl-1H-pyrazol-1-yl)-

Catalog No.:AG00066X MDL No.:

MF:C7H10N2O MW:138.1671

CAS No. 10199-64-1

Ethanone, 1-(1H-pyrazol-1-yl)-

Catalog No.:AG00066W MDL No.:

MF:C5H6N2O MW:110.1139

CAS No. 10199-66-3

Methanone, phenyl-1H-pyrazol-1-yl-

Catalog No.:AG00066V MDL No.:

MF:C10H8N2O MW:172.1833

CAS No. 10199-67-4

1H-Pyrazole, 1-(phenylmethyl)-

Catalog No.:AG00066U MDL No.:MFCD00462231

MF:C10H10N2 MW:158.1998

CAS No. 10199-68-5

1H-Pyrazole, 4-phenyl-

Catalog No.:AG00066T MDL No.:MFCD00462204

MF:C9H8N2 MW:144.1732

CAS No. 10199-69-6

1H-Pyrazole, 1-methyl-4-phenyl-

Catalog No.:AG00066S MDL No.:MFCD12964229

MF:C10H10N2 MW:158.1998

CAS No. 10199-85-6

Benzene, 1,3-dichloro-2,4-dinitro-

Catalog No.:AG00066R MDL No.:MFCD00823272

MF:C6H2Cl2N2O4 MW:236.9971

CAS No. 10199-87-8

1,3-Benzenediamine, 2,4-dinitro-

Catalog No.:AG00066Q MDL No.:

MF:C6H6N4O4 MW:198.1362

CAS No. 10199-89-0

2,1,3-Benzoxadiazole, 4-chloro-7-nitro-

Catalog No.:AG00066P MDL No.:MFCD00005808

MF:C6H2ClN3O3 MW:199.5514

CAS No. 10199-90-3

2,1,3-Benzoxadiazole, 4-azido-7-nitro-

Catalog No.:AG00066O MDL No.:

MF:C6H2N6O3 MW:206.1185

CAS No. 101990-17-4

Pyridine, 2-fluoro-5-[[2-(nitromethylene)-1-imidazolidinyl]methyl]-

Catalog No.:AG00066N MDL No.:

MF:C10H11FN4O2 MW:238.2183

CAS No. 101990-45-8

Pyridine, 2-bromo-5-(bromomethyl)-

Catalog No.:AG00066M MDL No.:MFCD11656267

MF:C6H5Br2N MW:250.9186

CAS No. 101990-64-1

3-Pyridinecarbonyl chloride, 6-(methylthio)-

Catalog No.:AG00066L MDL No.:

MF:C7H6ClNOS MW:187.6466

CAS No. 101990-68-5

3-Pyridinemethanol, 6-phenoxy-

Catalog No.:AG00066K MDL No.:MFCD02682069

MF:C12H11NO2 MW:201.2212

CAS No. 101990-69-6

4-Pyridinemethanol, 2,6-dichloro-

Catalog No.:AG00066J MDL No.:MFCD00052638

MF:C6H5Cl2NO MW:178.0160

CAS No. 101990-70-9

Pyridine, 5-(chloromethyl)-2-methoxy-

Catalog No.:AG00066I MDL No.:MFCD10697570

MF:C7H8ClNO MW:157.5975

CAS No. 101990-72-1

Pyridine, 2,6-dichloro-4-(chloromethyl)-

Catalog No.:AG00066H MDL No.:MFCD03783562

MF:C6H4Cl3N MW:196.4617

CAS No. 101990-73-2

Pyridine, 2-chloro-4-(chloromethyl)-

Catalog No.:AG00066G MDL No.:MFCD09991655

MF:C6H5Cl2N MW:162.0166

CAS No. 101991-76-8

1-Propanone, 2-(2,4-dichlorophenoxy)-1-(1-piperidinyl)-

Catalog No.:AG00066F MDL No.:MFCD01339369

MF:C14H17Cl2NO2 MW:302.1963

CAS No. 1019918-39-8

Pyridine, 4-bromo-2-hydrazinyl-

Catalog No.:AG00066E MDL No.:MFCD00234091

MF:C5H6BrN3 MW:188.0252

CAS No. 1019918-41-2

1,2,4-Triazolo[4,3-a]pyridin-3-amine, 7-bromo-

Catalog No.:AG00066D MDL No.:

MF:C6H5BrN4 MW:213.0347

CAS No. 1019918-43-4

1,2,4-Triazolo[4,3-a]pyridine, 7-bromo-3-(2-fluorophenyl)-

Catalog No.:AG000679 MDL No.:

MF:C12H7BrFN3 MW:292.1065

CAS No. 1019918-45-6

1,2,4-Triazolo[4,3-a]pyridine, 7-chloro-3-(2-fluorophenyl)-

Catalog No.:AG000678 MDL No.:

MF:C12H7ClFN3 MW:247.6555

CAS No. 1019918-47-8

1,2,4-Triazolo[4,3-a]pyridine, 7-bromo-3-(2-chlorophenyl)-

Catalog No.:AG000677 MDL No.:

MF:C12H7BrClN3 MW:308.5611

CAS No. 1019918-60-5

1,2,4-Triazolo[4,3-a]pyridine, 7-chloro-3-(2-chlorophenyl)-5-methyl-

Catalog No.:AG000676 MDL No.:

MF:C13H9Cl2N3 MW:278.1367

CAS No. 1019918-88-7

1,2,4-Triazolo[4,3-a]pyridine, 7-chloro-3-(2-chlorophenyl)-

Catalog No.:AG000675 MDL No.:

MF:C12H7Cl2N3 MW:264.1101

CAS No. 101992-41-0

Ethanedione, 1,1'-[1,6-hexanediylbis(oxy-4,1-phenylene)]bis[2-phenyl- (9CI)

Catalog No.:AG00067P MDL No.:

MF:C34H30O6 MW:534.5984

CAS No. 101992-47-6

Ethanedione, 1,1'-[1,3-propanediylbis(oxy-4,1-phenylene)]bis[2-phenyl- (9CI)

Catalog No.:AG00067O MDL No.:

MF:C31H24O6 MW:492.5187

CAS No. 101992-89-6

1-Butanol, 4-(methoxymethoxy)-

Catalog No.:AG00067N MDL No.:

MF:C6H14O3 MW:134.1736

CAS No. 1019929-89-5

Pyridine, 4,6-dichloro-3-methoxy-2-methyl-

Catalog No.:AG000674 MDL No.:MFCD11847534

MF:C7H7Cl2NO MW:192.0426

CAS No. 101993-02-6

2-Oxazolamine, 4,5-dihydro-5-(methoxymethyl)-

Catalog No.:AG00067M MDL No.:

MF:C5H10N2O2 MW:130.1451

CAS No. 1019930-14-3

2-Pyridinecarboxylic acid, 4-chloro-3-methoxy-6-phenyl-

Catalog No.:AG000673 MDL No.:

MF:C13H10ClNO3 MW:263.6764

CAS No. 101994-23-4

2H-1-Benzopyran-6-carbonitrile, 7-hydroxy-4,8-dimethyl-2-oxo-

Catalog No.:AG00067L MDL No.:

MF:C12H9NO3 MW:215.2048

CAS No. 101996-52-5

2H-Pyran, 3-chlorotetrahydro-2-(1-methylethyl)-, cis- (9CI)

Catalog No.:AG00067K MDL No.:

MF:C8H15ClO MW:162.6571

CAS No. 101997-49-3

Ethanone, 2-[[(2,3-dihydro-1,4-benzodioxin-2-yl)methyl]amino]-1-(1-piperidinyl)-

Catalog No.:AG00067J MDL No.:

MF:C16H22N2O3 MW:290.3575

CAS No. 101998-53-2

2-Propanone, 1-(1-methyl-2-imidazolidinylidene)-, (1E)-

Catalog No.:AG00067I MDL No.:

MF:C7H12N2O MW:140.1830

CAS No. 101999-40-0

2(3H)-Furanone, dihydro-3-(phenylmethoxy)-

Catalog No.:AG00067H MDL No.:

MF:C11H12O3 MW:192.2112

CAS No. 101999-44-4

2H-1-Benzopyran-2-one, 7-ethyl-4-methyl-

Catalog No.:AG00067G MDL No.:

MF:C12H12O2 MW:188.2225

CAS No. 101999-45-5

2H-1-Benzopyran-4-acetamide, 7-hydroxy-2-oxo-

Catalog No.:AG00067F MDL No.:MFCD06001840

MF:C11H9NO4 MW:219.1935

CAS No. 101999-46-6

2H-Naphtho[1,2-b]pyran-4-acetic acid, 2-oxo-

Catalog No.:AG00067E MDL No.:MFCD00434035

MF:C15H10O4 MW:254.2375

CAS No. 101999-59-1

1-Decen-3-one, 10-iodo-

Catalog No.:AG00067D MDL No.:

MF:C10H17IO MW:280.1459

CAS No. 101999-65-9

1,6-Tetradecadien-3-one, 14-iodo-, (6E)-

Catalog No.:AG00067C MDL No.:

MF:C14H23IO MW:334.2363

CAS No. 101999-70-6

4-Cyclotetradecen-1-one, (4E)-

Catalog No.:AG00067B MDL No.:

MF:C14H24O MW:208.3398

CAS No. 101999-73-9

Bicyclo[7.2.2]tridec-9(13)-ene

Catalog No.:AG00067A MDL No.:

MF:C13H22 MW:178.3138

CAS No. 1019995-16-4

3-Pyridinecarboxylic acid, 4-[(4-ethoxy-1,4-dioxobutyl)amino]-, ethyl ester

Catalog No.:AG000672 MDL No.:

MF:C14H18N2O5 MW:294.3031

CAS No. 1019995-18-6

1H-Pyrido[4,3-b]azepine-4-carboxylic acid, 2,3-dihydro-5-hydroxy-2-oxo-, ethyl ester

Catalog No.:AG00067R MDL No.:

MF:C12H12N2O4 MW:248.2347

CAS No. 1019996-86-1

1,1'-Biphenyl, 4-nitro-4'-(trifluoromethoxy)-

Catalog No.:AG00067Q MDL No.:MFCD21609608

MF:C13H8F3NO3 MW:283.2027

CAS No. 102-09-0

Carbonic acid, diphenyl ester

Catalog No.:AG000685 MDL No.:MFCD00003037

MF:C13H10O3 MW:214.2167

CAS No. 102-10-3

Phosphorous acid, diphenyl ester

Catalog No.:AG000684 MDL No.:

MF:C12H10O3P- MW:233.1798

CAS No. 102-11-4

1,2-Ethanediamine, N1,N2-dimethyl-N1-(phenylmethyl)-

Catalog No.:AG000683 MDL No.:MFCD00014856

MF:C11H18N2 MW:178.2740

CAS No. 102-13-6

Benzeneacetic acid, 2-methylpropyl ester

Catalog No.:AG000682 MDL No.:MFCD00026482

MF:C12H16O2 MW:192.2542

CAS No. 102-14-7

Butanoic acid, 4-oxo-4-(phenylamino)-

Catalog No.:AG000681 MDL No.:MFCD00029825

MF:C10H11NO3 MW:193.1992

CAS No. 102-16-9

Benzeneacetic acid, phenylmethyl ester

Catalog No.:AG000680 MDL No.:MFCD00022045

MF:C15H14O2 MW:226.2705

CAS No. 102-17-0

Benzeneacetic acid, (4-methoxyphenyl)methyl ester

Catalog No.:AG00067Z MDL No.:

MF:C16H16O3 MW:256.2964

CAS No. 102-19-2

Benzeneacetic acid, 3-methylbutyl ester

Catalog No.:AG00067Y MDL No.:MFCD00026517

MF:C13H18O2 MW:206.2808

CAS No. 102-20-5

Benzeneacetic acid, 2-phenylethyl ester

Catalog No.:AG00067X MDL No.:MFCD00022049

MF:C16H16O2 MW:240.2970

CAS No. 102-22-7

Benzeneacetic acid, (2E)-3,7-dimethyl-2,6-octadien-1-yl ester

Catalog No.:AG00067W MDL No.:MFCD00072225

MF:C18H24O2 MW:272.3820

CAS No. 102-23-8

Benzenesulfonic acid, 3-[2-(4-aminophenyl)diazenyl]-

Catalog No.:AG00067V MDL No.:

MF:C12H11N3O3S MW:277.2990

CAS No. 102-24-9

Boroxin, 2,4,6-trimethoxy-

Catalog No.:AG00067U MDL No.:MFCD00005946

MF:C3H9B3O6 MW:173.5330

CAS No. 102-25-0

Benzene, 1,3,5-triethyl-

Catalog No.:AG00067T MDL No.:MFCD00009261

MF:C12H18 MW:162.2713

CAS No. 102-26-1

1,3,5-Triazine, 2,4,6-triethylhexahydro-

Catalog No.:AG00067S MDL No.:

MF:C9H21N3 MW:171.2831

CAS No. 16530-72-6

Octanoic acid, compd. with 2,2'-iminobis[ethanol] (1:1)

Catalog No.:AG000648 MDL No.:

MF:C12H27NO4 MW:249.3471

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