Startseite Synthesis, antibacterial, and antifungal activities of new pyrimidinone derivatives
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Synthesis, antibacterial, and antifungal activities of new pyrimidinone derivatives

  • Oussama Cherif , Fatma Masmoudi , Fatma Allouche , Fakher Chabchoub EMAIL logo und Mohamed Trigui
Veröffentlicht/Copyright: 23. Juli 2015
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Heterocyclic Communications
Aus der Zeitschrift Heterocyclic Communications Band 21 Heft 4

Abstract

An efficient synthesis of new pyrrolopyrimidinones 3a-d and isoxazolopyrimidinones 4a-c from the respective aminocyanopyrroles 1a-d and aminocyanoisoxazoles 2a-c is presented. The synthesized compounds were screened for antimicrobial activity against a panel of bacteria and fungi. Compound 4c exhibits remarkable activity against a broad spectrum of Gram-positive and Gram-negative bacteria and pathogenic fungi.

Keywords: antimicrobial activity; isoxazolopyrimidinones; pyrrolopyrimidinones

Pyrroles and isoxazoles [1–4] have received intensive research interests because of their biological activities and have found a wide range of applications in pharmaceutical and agrochemical fields [5–8]. In continuation of our previous study on the synthesis and biological activity of heterocyclic compounds [9], we now report the preparation of a series of pyrrolopyrimidinones 3a-dand isoxazolopyrimidinones 4a-cstarting with readily available pyrroles 1a-d and isoxazoles 2a-c [10, 11]. Compounds 3a-d and 4a-c were synthesized by heating the respective substrates 1a-d and 2a-c under reflux in formic acid in the presence of a catalytic amount of sulfuric acid. The yields ranged from 57% to 81% (Scheme 1). Mechanistically, the reaction may involve the partial hydrolysis of the cyano group to give an amide and the formylation of the amino group to give a formamide followed by intramolecular condensation of these two functionalities, as shown.

Scheme 1
Scheme 1

All compounds were screened for their antibacterial and antifungal activities against a number of reference test organisms, including five Gram-positive, three Gram-negative, and eight plant pathogenic fungi. The study of antimicrobial activities showed that compounds 1–3 and 4a,b exhibit very low activities. A notable exception is compound 4c, which is active against a broad spectrum of Gram-positive and Gram-negative bacteria (Table 1). The same compound 4c shows also high antifungal activity (Table 2).

Table 1

Antibacterial activities of compound 4cin vitro.

Bacterial strains
Gram-positive bacteriaGram-negative bacteria
BcSaEfMlLmEcSeKp
Compound 4cIZa1810130912111215
MICb0.010.0430.0850.0850.6870.0850.1720.085
GentamicincIZ2020201820252522
MIC0.0040.0040.0040.0040.0010.0020.0020.002
DMSO 20%IZd––––––––

Bacterial strains: Bc, Bacillus cereus ATCC14579; Sa, Staphylococcus aureus ATCC25923; Ef, Enterococcus faecalis ATCC 29212; Ml, Micrococcus luteus ATCC 1880; Lm, Listeria monocytogenes (FI 2132); Ec, Escherichia Coli ATCC 25922; Se, Salmonella enteritidis (food isolate); Kp, Klebsiella pneumonia CIP 32147.

aDiameter of the inhibition zone (IZ) in mm of a compound, including the diameter of the disc (6 mm).

bMinimum inhibitory concentration in mg/mL.

cGentamicin was used as a standard antibiotic at 15 μg/mL.

dActivity not detected.

Table 2

Antifungal activities of compound 4c.

CompoundsFungal strains
RnFoAaPpFcBcFgAf
4cIZa1110131213151019
MICb0.6870.1720.6870.3430.1720.6870.3435.5
AmphotericincIZ1012121814121410
MIC0.3430.0420.1720.3120.0850.1720.3430.172
DMSO 20%IZd––––––––

Fungal strains: Rn, Rhizopus nigricans (LPAP26); Fo, Fusarium oxysporum CTM10402; Aa, Alternaria alternata CTM 10230; Pp, Pythium aphanidermatum (LPAP32); Fc, Fusarium culmorum ISPAVE 21W; Bc, Botrytis cinerea (LPAP34); Fg, Fusarium graminearum ISPAVE 271; Af, Aspergillus flavus (food isolate).

aDiameter of the inhibition zone of a compound, including the diameter of the disc (6 mm).

bMinimal inhibitory concentration in mg/mL.

cAmphotericin B was used as antifungal standard at 20 μg/mL.

dActivity not detected.

Experimental

Melting points were measured on an electrothermal apparatus. Progress of the reactions was monitored by thin-layer chromatography (TLC) using aluminum sheets with silica gel 60 F254 from Merck. Infrared spectra (IR) were recorded on a Perkin-Elmer Paragon FT-IR spectrometer using KBr pellets. Unless stated otherwise, 1H NMR and 13C NMR spectra were recorded in CDCl3 on a Bruker spectrometer (1H at 400 MHz, 13C at 100 MHz). Elemental analyses were performed on a Carlo Erba 1106 apparatus. Substrates 1a-d and 2a-c were synthesized as previously reported [10, 11].

General procedure for the synthesis of compounds 3a-d and 4a-c

3-Amino-4-cyano-substituted pyrrole 1or 5-amino-4-cyano-substituted isoxazole 2(10 mmol) was added portionwise for 2 h to a mildly refluxing mixture of formic acid (20 mL) and concentrated sulfuric acid (1.2 mL). After an additional 30 min, the solution was cooled to 0°C and poured onto crushed ice. The resulting precipitate was collected by filtration, washed with water, dried, and crystallized from ethanol to give a pyrrolopyrimidinone 3 or an isoxazolopyrimidinone 4.

Methyl 5,6-dimethyl-4-oxo-4,6-dihydro-3H-pyrrolo[3,4-d]pyrimidine-7-carboxylate (3a)

Yield 81%; yellow solid; mp 231–233°C; IR (cm-1): ν 1596, 1692, 3187; 1H NMR: δ 8.29 (s, 1H, NH), 7.89 (s, 1H, H-2), 4.01 (s, 3H, CH3-7), 2.76 (s, 3H, CH3-6), 2.19 (s, 3H, CH3-5); 13C NMR: δ 159.3 (C-4), 157.7 (CO2Me), 145.2 (C-2), 143.6 (C-5), 133.9 (C-7a), 114.1 (C, C-7), 106.6 (C-4a), 31.8 (CH3-7), 13.3 (CH3-6), 10.6 (CH3-5). Anal. Calcd for C10H11N3O3: C, 54.29; H, 5.01; N, 19. Found: C, 53.80; H, 5.20; N, 18.33.

Ethyl 5,6-dimethyl-4-oxo-4,6-dihydro-3H-pyrrolo[3,4-d]pyrimidine-7-carboxylate (3b)

Yield 71%; yellow solid; mp 243–245°C; IR (cm-1): ν 1597, 1691, 3173; 1H NMR: δ 8.24 (s, 1H, NH), 8.12 (s, 1H, H-2), 4.43 (q, 2H, 3J = 7.5 Hz, CH2), 3.91 (s, 3H, CH3-6), 2.61 (s, 3H, CH3-5), 1.33 (t, 3H, 3J = 7.5 Hz, CH3-7); 13C NMR: δ 160.1 (C-4), 159.1 (CO2Et), 146.2 (C-2), 144.9 (C-5), 135.3 (C-7a), 115.4 (C-7), 107.0 (C-4a), 59.9 (CH2), 32.9 (CH3-6), 14.4 (CH3-7), 11.0 (CH3-5). Anal. Calcd for C11H13N3O3: C, 56.16; H, 5.57; N, 17.86. Found: C, 55.66; H, 6.02; N, 17.38.

Methyl 5-ethyl-6-methyl-4-oxo-4,6-dihydro-3H-pyrrolo[3,4-d] pyrimidine-7-carboxylate (3c)

Yield 73%; yellow solid; mp 247–250°C; IR (cm-1): ν 1597, 1680, 3187; 1H NMR: δ 8.32 (s, 1H, NH), 8.11 (s, 1H, H-2), 4.09 (s, 3H, CH3-7), 4.01 (s, 3H, CH3-6), 3.23 (q, 2H, 3J = 7.5 Hz, CH2), 1.29 (t, 3H, 3J = 7.5 Hz, CH3-5); 13C NMR: δ 160.7 (C-4), 158.2 (CO2Me), 145.8 (C-2), 141.2 (C-5), 139.5 (C-7a), 111.3 (C-7), 106.0 (C-4a), 51.0 (CH3-7), 32.9 (CH3-6), 17.5 (CH3-5), 13.1 (CH2). Anal. Calcd for C11H13N3O3: C, 56.16 H, 5.57; N, 17.86 . Found: C, 55.04; H, 5.83; N, 17.29.

Methyl 6-methyl-4-oxo-5-phenyl-4,6-dihydro-3H-pyrrolo[3,4-d]pyrimidine-7carboxylate (3d)

Yield 65%; yellow solid; mp 232–235°C; IR (cm-1): ν 1600, 1693, 3296; 1H NMR: δ 8.34 (s, 1H, NH), 8.13 (s, 1H, H-2), 7.46–7.56 (m, 5H, H-arom), 4.14 (s, 3H, CH3-7), 4.01 (s , 3H, CH3-6); 13C NMR: δ 161.1 (C-4), 157.7 (CO2Me), 145.4 (C-2), 141.4 (C-5), 127.9–136.9 (C-arom), 115.3 (C-7a), 114.4 (C-7), 107.5 (C-4a), 51.1 (CH3-7), 35.0 (CH3-6). Anal. Calcd for C15H13N3O3: C, 63.60; H, 4.63; N, 14.83. Found: C, 62.76; H, 4.64; N, 14.29.

3-Methylisoxazolo[5,4-d]pyrimidin-4(5H)-one (4a)

Yield 66%; white solid; mp 232–235°C; IR (cm-1): ν 1567, 1707, 3182; 1H NMR: δ 12.74 (s, 1H, NH), 8.28 (s, 1H, H-6), 2.46 (s, 3H, CH3); 13C NMR: δ 175.10 (C-7a), 157.7 (C-4), 157.2 (C-3), 152.2 (C-6), 101.5 (C-3a), 10.9 (CH3). Anal. Calcd for C6H5N3O2: C, 47.69; H, 3.33; N, 27.81. Found: C, 46.83; H, 4.16; N, 27.05.

3-ethylisoxazolo[5,4-d]pyrimidin-4(5H)-one (4b)

Yield 61%; white solid; mp 185–188°C; IR (cm-1): ν 1567, 1702, 3188; 1H NMR: δ 12.21 (s, 1H, NH), 8.24 (s, 1H, H-6), 3.03 (q, 2H, 3J = 7.7 Hz, CH2), 1.45 (t, 3H, 3J = 7.7 Hz, CH3); 13C NMR: δ 175.9 (C-7a), 158.9 (C-4); 157.8 (C-3), 154.2 (C-6), 102.7 (C-3a), 15.9 (CH2), 10.6 (CH3). Anal. Calcd for C7H7N3O2: C, 50.91; H, 4.27; N, 25.44. Found: C, 50.95; H, 4.37; N, 25.37.

3-Phenylisoxazolo[5,4-d]pyrimidin-4(5H)-one (4c)

Yield 57%; white solid; mp 216–218°C; IR (cm-1): ν 1559, 1692, 3190; 1H NMR: δ 10.58 (s, 1H, NH), 8.12 (s, 1H , H-6), 7.19–8.24 (m, 5H, H-arom); 13C NMR: δ 170.5 (C-7a), 161.2 (C-4) , 160.2 (C-3), 144.7 (C-6), 123.4–130.7 (C-Carom), 115.4 (C-3a). Anal. Calcd for C11H7N3O2: C, 61.97; H, 3.31; N, 19.71. Found: C, 61.32; H, 3.68; N, 18.69.

Antimicrobial activity

Eight bacterial and eight fungal strains were used. The tested pathogenic bacteria were Bacillus cereus ATCC 14579 (Bc), Staphylococcus aureus ATCC 25923 (Sa), Enterococcus faecalis ATCC 29212 (Ef), Micrococcus luteus ATCC 1880 (Ml), Escherichia coli ATCC 25922 (Ec), Klebsiella pneumoniae ATCC 10031 (Kp), Salmonella enteritidis (food isolate 824) (Se), and Listeria monocytogenes (food isolate 2132) (Lm). The fungi tested were Rhizopus nigricans (LPAP26) (Rn), Fusarium oxysporum CTM10402 (Fo), Alternaria alternata CTM 10230 (Aa), Pythium aphanidermatum (LPAP32) (Pp), Fusarium culmorum ISPAVE 21W (Fc), Botrytis cinerea (LPAP34) (Bc), Fusarium graminearum ISPAVE 271 (Fg), and Aspergillus flavus (food isolate) (Af). The bacterial strains were cultivated in Mueller-Hinton (MH) agar (Oxoid Ltd., UK) at 37°C except for Bacillus species, which were incubated at 30°C. The fungi were cultured on potato dextrose agar (PDA) medium and incubated at 28°C. Working cultures were prepared by inoculating a loopful of each test bacteria in 3 mL of MH broth (Oxoid Ltd., UK) and were incubated at 37°C for 12 h. For the test, final inoculum concentrations of 107 CFU/mL of bacteria were used. Fungal spore suspensions were collected from the surface of such fungal colonies by gently scraping with a loop and suspended in 10 mL potato dextrose broth. This suspension was mixed vigorously by vortexing for 15–20 min. The spore suspension stock was diluted to obtain a concentration of 106 spores/mL (measured by Malassez blade). Antibacterial and antifungal tests were performed by disc diffusion method [12] and broth microdilution assay using sterile MH media (Bio-Rad, France) for bacterial strains PDA (Bio-Rad, France) for antifungal tests. Freshly prepared cell suspension (100 μL) adjusted to 107 CFU/mL for bacteria and 106 spores/mL for fungus were inoculated onto the surface of agar plates. Thereafter, discs with 6 mm in diameter were punched in the inoculated agar medium with sterile Pasteur pipettes, and compounds were added to each disc. Gentamicin (10 μg/disc) was used as a positive control for bacteria, whereas amphotericin B (20 μg/disc) was used as a positive control for fungal strains. The plate was allowed to stand for 2 h at 4°C to permit the diffusion of the compounds followed by incubation at 37°C for 24 h for bacterial strains and 72 h for fungi at 28°C. The antibacterial activity was evaluated by measuring the zones of inhibition (clear zone around the disc) against the tested microorganisms. All tests were repeated three times. Minimum inhibitory concentrations (MICs) of compounds were determined in sterile 96-well microplates with a final volume in each microplate well of 200 μL [13]. A twofold serial dilution of the compound was prepared in the microplate wells over the range 0.01–5.5 mg/mL. To each test well was added 10 μL cell suspension to final inoculum concentrations of 106 CFU/mL for bacteria and 105 spores/mL for fungi. The plates were then covered with the sterile plate covers and incubated at 37°C for 24 h for bacterial strains and 72 h for fungi at 28°C. The MIC was defined as the lowest concentration of the compound at which the microorganism does not demonstrate visible growth after incubation. As an indicator of microorganism growth, 25 μL of 0.5 mg/mL p-iodonitrotetrazolium chloride, dissolved in sterile water, was added to the wells and incubated at 37°C for 30 min. The lowest concentration of compound showing no growth was taken as its MIC.

Corresponding author: Fakher Chabchoub, Laboratory of Applied Chemistry Heterocycles, Fats and Polymers, Faculty of Sciences of Sfax, University of Sfax, Sfax 3018, Tunisia, e-mail:

Acknowledgments

The authors acknowledge the Ministry of Higher Education, Scientific Research, and Technology in Tunisia for their financial support.

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Received: 2015-3-6
Accepted: 2015-4-12
Published Online: 2015-7-23
Published in Print: 2015-8-1

©2015 by De Gruyter

This article is distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Artikel in diesem Heft

  1. Frontmatter
  2. Preliminary Communications
  3. Montmorillonite K10 catalyzed multi component reactions (MCR): synthesis of novel thiazolidinones as anticancer agents
  4. Synthesis, antibacterial, and antifungal activities of new pyrimidinone derivatives
  5. Research Articles
  6. Formation of 1-methyl[1,2,4]triazolo[4,3-a] quinazolin-5(4H)-ones by reaction of 2-hydrazinoquinazolin-4(3H)-ones with acetylacetone
  7. Synthesis of new 4′-(N-alkylpyrrol-2-yl)-2,2′: 6′,2″-terpyridines via N-alkylation of a pyrrole moiety
  8. Efficient synthesis of 3-(bromomethyl)-5-methylpyridine hydrobromide
  9. One-pot synthesis of 5-[1-substituted 4-acetyl-5-methyl-1H-pyrrol-2-yl)]-8-hydroxyquinolines using DABCO as green catalyst
  10. A new on-fluorescent sensor for Ag+ based on benzimidazole bearing bis(ethoxycarbonylmethyl)amino groups
  11. Synthesis of new derivatives of 10H-benzo[b]pyridazino[3,4-e][1,4]thiazines
  12. Efficient and convenient synthesis of pyrido [2,1-b]benzothiazole, pyrimidopyrido[2,1-b]benzothiazole and benzothiazolo[3,2-a][1,8]naphthyridine derivatives
  13. Synthesis of 3-benzylidene-dihydrofurochromen-2-ones: promising intermediates for biflavonoid synthesis
  14. Synthesis and antitumor activities of piperazine- and cyclen-conjugated dehydroabietylamine derivatives
  15. Synthesis, characterization, and antimicrobial evaluation of novel spiropiperidones
https://doi.org/10.1515/hc-2015-0066
Schlagwörter für diesen Artikel
antimicrobial activity; isoxazolopyrimidinones; pyrrolopyrimidinones
Creative Commons
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Artikel in diesem Heft

  1. Frontmatter
  2. Preliminary Communications
  3. Montmorillonite K10 catalyzed multi component reactions (MCR): synthesis of novel thiazolidinones as anticancer agents
  4. Synthesis, antibacterial, and antifungal activities of new pyrimidinone derivatives
  5. Research Articles
  6. Formation of 1-methyl[1,2,4]triazolo[4,3-a] quinazolin-5(4H)-ones by reaction of 2-hydrazinoquinazolin-4(3H)-ones with acetylacetone
  7. Synthesis of new 4′-(N-alkylpyrrol-2-yl)-2,2′: 6′,2″-terpyridines via N-alkylation of a pyrrole moiety
  8. Efficient synthesis of 3-(bromomethyl)-5-methylpyridine hydrobromide
  9. One-pot synthesis of 5-[1-substituted 4-acetyl-5-methyl-1H-pyrrol-2-yl)]-8-hydroxyquinolines using DABCO as green catalyst
  10. A new on-fluorescent sensor for Ag+ based on benzimidazole bearing bis(ethoxycarbonylmethyl)amino groups
  11. Synthesis of new derivatives of 10H-benzo[b]pyridazino[3,4-e][1,4]thiazines
  12. Efficient and convenient synthesis of pyrido [2,1-b]benzothiazole, pyrimidopyrido[2,1-b]benzothiazole and benzothiazolo[3,2-a][1,8]naphthyridine derivatives
  13. Synthesis of 3-benzylidene-dihydrofurochromen-2-ones: promising intermediates for biflavonoid synthesis
  14. Synthesis and antitumor activities of piperazine- and cyclen-conjugated dehydroabietylamine derivatives
  15. Synthesis, characterization, and antimicrobial evaluation of novel spiropiperidones
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