SYNTHESIS, CHARACTERIZATION AND ANTIMICROBIAL EVALUATION OF2,3-DIHYDRO-1H-PERIMIDINE DERIVATIVES

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50

Journal Marocain de Chimie Hétérocyclique Moroccan Journal of Heterocyclic Chemistry

ISSN : 2605-5996

J. Mar. Chim. Heterocycl., 2021,Volume 20, Issue 3, Page 50-56

SYNTHESIS, CHARACTERIZATION AND ANTIMICROBIAL EVALUATION OF2,3-DIHYDRO-1H-PERIMIDINE DERIVATIVES.

TUO Nanou Tiéba

¹

, KANGAH Niameke Jean Baptiste

¹

, BALLO Daouda

^2,3

, SANHOUN Aimé R.

⁴

, Kablan Ahmont Landry Claude

⁵

, KODJO Charles Guillaume

¹

, YAPO Ossey Bernard

^6,7

and ZIAO Nahossé

¹

1Laboratoire de Thermodynamique et de Physico-Chimie du Milieu, UFR SFA, Université Nangui Abrogoua 02 BP 801 Abidjan 02, Côte-d’Ivoire,

2Laboratoire de Chimie Organique Heterocyclique URAC 21, Pôle de Compétence Pharmacochimie, Faculté des Sciences, Universite´ Mohammed V, Rabat, Moroc,

3Laboratoire de Constitution et Réaction de la Matière (LCRM), UFR Sciences des Structures de la Matière et Technologie, Université Félix Houphouët-Boigny, 22 BP 582 Abidjan, Côte d’Ivoire,

4Centre Suisse de Recherche Scientifiques en Côte d’Ivoire (CSRS),

5UFR Des Sciences Biologiques, Université Péléforo Gon Coulibaly de Korhogo, BP 1328 Korhogo, Côte d’Ivoire,

6Laboratoire Centrale de l’Environnement (LCE), Centre Ivoirien Antipollution (CIAPOL), 20 BP 650 Abidjan 20, Côte d’Ivoire,

7Laboratoire des Sciences de l’Environnement (LSE), UFR SGE, Université Nangui Abrogoua 02 BP 801 Abidjan 02, Côte-d’Ivoire.

Received ; 05/03/2021 ; Accepted 24/06/2021

ABSTRACT

We report here the synthesis, characterization and antimicrobial activity of three molecules from the family of 2,3-dihydro -1H-perimidines derived from 1,8-diaminonaphthalene. The three perimidines were characterized by conventional spectrometry methods (NMR and MS), then tested on bacterial strains. The three compounds 1-3 were shown to be active against the strain of bacteria Escherichia coli ATCC 25922, with MIC values of 7.5 mg / mL, 3.75 mg / mL and 60 mg / mL respectively. Salmonella typhimirium SO66 is active on compounds 1 and 2 at MIC values of 30 mg / mL and 60 mg / mL, respectively. Compound 1 is active on Staphylococcus aureus CIP 4,83 and Staphylococcus epidermidice CIP 54124 at MIC values of 7.5 mg / mL and 1.87 mg / mL and for compound 2 we have 3.75 mg / mL and 0.225 mg / mL respectively. The fungal strain of Candida Albicans ATCC 10234 showed resistance to compound 3 synthesized, but on the other hand Candida tropicalis ATCC 13803 was sensitive to all compounds with a MIC of 0.45 mg / ml for compound 1, 0.225 mg / ml for compound 2 and 3.75 mg / ml for compound 3.

Therefore, the para position appears to be the least active position on the antimicrobial sources of this pharmacophore.

KEYWORDS: 1H-Perimidine, spectrometry, antimicrobial activity,

inhibition

and

1,8- Diaminonaphthalene

.

Tuo, J. Mar. Chim. Heterocycl., 2021, Volume 20, Issue 3, Page 50-56

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51 Introduction

Bacterial infections remain the most harmful to human health [1 - 2]. There is therefore an urgent need to develop new chemical entities that are more effective than those currently available on the market, thus making it possible to propose possible solutions to the problems of drug resistance [3; 4]. 2,3-Dihydro-1H-perimidines are obtained through the condensation of primary amines with carbonyl compounds. These are six-membered heterocyclic compounds bearing two nitrogen atoms that have been extensively studied [5 - 9]. They are characterized by either a binding deficit or an excess of Π binding [10] and are of great interest due to their biological activities [11], in particular their potential to act as antimicrobial and antifungal agents [12 -14]. As part of our research on 2,3-dihydro -1H- perimidines, we are interested in their biological activities

Thus, this article discusses the synthesis, characterization, and biological studies of all compounds synthesized from the reagents shown in Table 1.

Table 1: Structures of

2,3-Dihydro-1H-perimidines

synthesized.

compound Ar

1

2

3 RESULTS AND DISCUSSION

The condensation of diamino-naphtalene with benzaldehyde derivatives are carried out in refluxing ethanol to afford compounds 1 and 3 or at room temperature in ether leading to compound 2 The general synthesis of the the three compounds is shown in scheme 1.

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Scheme 1: The general synthesis route of compounds 1–3.

The structures of compounds 1-3 have been elucidated on the basis of spectral data ( NMR and Mass Spectrometry)

1H NMR and ¹³C NMR Spectroscopy

1H NMR and ¹³C NMR spectral data in deturated CDCl3 solution of the synthesized compounds are given in Table 2. The resonance of protons had been assigned on the basis of their integration and multiplicity pattern [24]. The ¹H NMR spectra exhibited signals at 3.51;

3.50; 3.36 ppm respectively for compounds 1, 2 and 3, attributed to protons bonded to azote atoms CH-N-. The singulet at 5.80; 5.75; 5.29 ppm for compounds 1, 2 and 3, attributed to perimidine cyclic protons CH-, respectively. The multi-signals within the 8.19-6.47 ppm range are assigned to the aromatic protons of the three heterocyclic compounds.

The signals at 60.97; 61.46; 76.6 ppm are attributed to carbon bonded both to two azote atoms and to phenyl group.

Table 2:

¹

H NMR data

^a-c

and

¹³

C NMR data of compounds compounds Molecular

formula

N-H m

C-H s

C

6

-H m

N-C-H 1 C

₁₇

H

₁₄

N

₂

3.51(2H) 5.80 (1H) 8.08 - 6.47 (11H) 60.97 2 C

17

H

14

N

2

O 3.50 (2H) 5.75 (1H) 8.24 - 6.48 (10H) 61.46 3 C

17

H

14

N

2

O 3.36 (2H) 5.29 (1H) 8.19 - 6.64 (10H) 76.6

a Multiplicity is given as s = singlet, m = multi-signals b Chemical shits in ppm

c Integration: number of protons in brackets

The 1H-NMR spectral data of the perimidines synthesized were in accord with the proposed structures

.

MS study

The mass spectra (HR-ESI-MS) of the title compounds showed peaks corresponding to the protonated molecular ions at m/z 263 [M + H]⁺, that allowed to propose C17H14N2O empirical formula for compounds 2 and 3. Concerning compounds 1 the peak at m/z 247 [M + H]⁺, was conform to propose C₁₇H₁₄N₂ empirical formula.

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Biological Activity

Antibacterial activity

The results of the antibacterial screening of compounds 1, 2 and 3 at a concentration of 60 mg / ml against Escherichia coli ATCC 25922, Salmonella typhimirium SO66, Staphylococcus aureus CIP 4,83, and Staphylococcus epidermidice CIP 54124. The results of the table show that compounds 1 an d 2 are sensitive against all the strains tested. Compound 3 is shown to be the most active against Escherichia coli ATCC 25922 with an MIC of 60 mg / mL, but insensitive against Salmonella typhimirium SO66 and Staphylococcus aureus CIP 4,83. The zones of inhibition were measured in mm and the results are shown in Table 3. The diameters of the zones of inhibition were between 10 and 20 mm.

Table 3: Mean diameters (mm) of the inhibition zones and Value of Minimum Inhibitory Concentration (MIC) values for antibacterial activity

Mean diameters of the inhibition zones (mm) Value of MIC (mg/mL) Souches

testées

E. coli Sal.

typhi

Sta.

aureus

Sta. epid.

E. coli

Sal.

typhi

Sta.

aureus Sta.

epid.

Concentrations (mg / mL) C1= 60, C2= 15

Composés C1 C2 C1 C2 C1 C2 C1 C2

1 16 14 10 0 15 12 14 10 7,5 30 7,5 1,87

2 20 19 14 0 21 20 14 12 3,75 60 3,75 0,225

3 10 0 0 0 0 0 0 0 60 - - 60

Witnesses

Gen. 0 0 0 0 25 0 30 29 0.007 0.007 0.007 0,014

Les valeurs sont des moyennes de trois répétitions ; Gen : Gentamicin, E. Coli : Escherichia coli, Sal. Typhi : Salmonella typhi, Sta. Aureus : Staphylococcus aureus, Sta. Epid. : Staphylococcus epidermidice.

Antifungal activity

Amphotericin B and compound 3 show no antifungal activity against Candida albicans ATCC 10234, but show activity on Candida tropicalis ATCC 13803 with diameters of inhibitions between 12 mm and 12 mm for compound 3 and between 29.5 mm and 30 mm for amphotericin B. compounds 1 and 2 are shown to be sensitive against the two strains tested with inhibition diameters ranging from 9 mm to 17 mm as indicated in Table 4.

However, on Candida tropicalis ATCC 13803 compound 3 is the most active with a MIC value of 3.75 mg / mL.

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Table 4: Mean diameters (mm) of the inhibition zones and Value of Minimum Inhibitory Concentration (MIC) values for antifungal activity

Les diamètres moyens des zones d’Inhibition (mm) Valeurs des CMI (mg / mL) Souches

testées

Candida albicans Candida tropicalis

Candida albicans

Candida tropicalis Concentrations (mg / mL)

C1= 60, C2=15

Composés C1 C2 C1 C2

1 12 10 10 09 3,75 0,45

2 17 15 15 14 3,75 0,225

3 - - 12 11 - 3,75

Witnesses

Amp. B 0 0 30 29,5 - 0,014

Les valeurs sont des moyennes de trois répétitions ; Amp. B : Amphotericine B

Experimental

Benzaldehyde, salicylaldehyde, 4-hydroxybenzaldehyde, and benzene-1,8-diaminonaphtalène were procured from Aldrich and used without further purification. All organic solvents were purchased from Merck and dried before use. Melting points were determined in capillary tube using an MPD Mitamura Riken Kogyo (Japan) electrothermal melting point apparatus and are uncorrected. The 1H NMR spectra were recorded on a Bruker-Avance-300 spectrometer, operating at 300 MHz. The mass spectra were recorded on a TOF LCT Premier (WATERS) Spectrometer coupled to an HPLC Alliance 2695 chain.

Synthesis of 2-phenyl-2,3-dihydro-1H-perimidine 1

Benzaldehyde (19.60 mmol) and 1,8-diaminonaphtalène (9.79 mmol) were dissolved in ethanol (50 ml). The mixture was heated at reflux for 7 hours. After cooling to the green precipitate obtained was filtered and rinsed with ethanol (Rf: 0.87 in hexane/acetate d’ethyle (2;1), yield: 82%, mp: 206^oC).

Synthesis of 2-(2,3-dihydro-1H-perimidin-2-yl)phenol 2

salicylaldehyde (18.80 mmol) and 1,8-diaminonaphtalène (9.35 mmol) were dissolved in ether (30 ml). The mixture was stirred at room temperature for three days to give a brown precipitate. which was filtered and rinsed with ether (Rf: 0.70 in hexane /acetate d’ethyle (2;1) yield: 27%, mp: 198.2^oC).

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Synthesis of 4-(2,3-dihydro-1H-perimidin-2-yl)phenol 3

4-Hydroxybenzaldehyde (12.61 mmol) and 1,8-diaminonaphtalène (6.32 mmol) were dissolved in ethanol (50 ml). The mixture was heated at reflux for 5 hours to give a maroon precipitate which was filtered and rinsed with ethanol (Rf: 0.61 in hexane /acetate d’ethyle (2;1), yield: 48%, mp >268 °C).

Biological activity Antibacterial testing

The bacterial cultures: Escherichia coli ATCC 25922, Salmonella typhimirium SO66, Staphylococcus aureus CIP 4,83, and Staphylococcus epidermidice CIP 54124 were obtained from Microbiology Laboratory of Swiss Scientific Research Center of Abidjan (Cote D’Ivoire). The bacterial cultures were incubated at 37°C for 18 hours by inoculation into nutrient agar. Schiff bases were stored dry at room temperature and dissolved 60 mg/mL then 3,75 mg/ml in dimethylsulfoxide (DMSO). Antibacterial activities of each compound were evaluated by the agar disc-diffusion method. Mueller Hinton Agar Media (15 cm3) kept at 45^oC was poured in the Petridishes and allowed to solidify. Poured Petri plates (9 cm) were incubated with 50µL of normal saline solution of above culture media (105-106 bacteria per ml). Discs injected with prepared perimidine (50µl) were applied on the solid agar medium by pressing tightly. The Petri plates were placed at 37^oC for 18 hours. At the end of period the inhibition zones formed on media were measured with a zone reader in millimeters.

Antifungal testing

Pathogenic strains of Candida albicans ATCC 10234 and Candida tropicalis ATCC 13803 were obtained from the Microbiology Laboratory of Swiss Scientific Research Center in Cote D’Ivoire. Perimidines were stored dry at room temperature and dissolved 60mg/mL in dimethylsulfoxide (DMSO). Antifungal activities of each compound were evaluated by the agar disc-diffusion method. Sabarod’s agar media (15 cm3) kept at 45^oC was poured in the Petri-dishes and allowed to solidify. Sterile, filter paper discs of 10mm diameter were impregnated with prepared perimidine (50µl) and were placed on to the media, seeded with fungus. The plates were then incubated at 37^oC for 1-3 days. At the end of period the inhibition zones formed on media were measured with a zone reader in millimeters.

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56 Conclusion

We can therefore retain that our compounds have bacterial activity and are all sensitive against Escherichia coli ATCC 25922 and Staphylococcus epidermidice CIP 54124.

Compound 2 is shown to be the most active on Escherichia coli ATCC 25922 and Staphylococcus epidermidice CIP 54124, with MIC values of 3.75 mg / mL and 0.225 mg / mL, respectively. The antifungal assays have shown that our compounds are all active against Candida albicans ATCC 10234 with a MIC value of 3.75 mg / mL, except compound 3 which shows resistance. Regarding Candida tropicalis ATCC 13803, it is sensitive to all three compounds and the compound is the most active with a MIC value of 0.225 mg / mL.

However, 2,3-dihydro -1H-perimidine are biologically active.

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