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A Convenient Method for the Synthesis of (Prop-2-Ynyloxy)Benzene Derivatives via Reaction with Propargyl Bromide, Their Optimization, Scope and Biological Evaluation

A Convenient Method for the Synthesis of (Prop-2-Ynyloxy)Benzene Derivatives via Reaction with Propargyl Bromide, Their Optimization, Scope and Biological Evaluation

  • Tannaza Batool, 
  • Nasir Rasool, 
  • Yasmeen Gull, 
  • Mnaza Noreen, 
  • Faiz-ul-Hassan Nasim, 
  • Asma Yaqoob, 
  • Muhammad Zubair, 
  • Usman Ali Rana, 
  • Salah Ud-Din Khan, 
  • M. Zia-Ul-Haq
PLOS
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Abstract

A highly convenient method has been developed for the synthesis of (prop-2-ynyloxy) benzene and its derivatives. Differently substituted phenol and aniline derivatives were allowed to react with propargyl bromide in the presence of K2CO3 base and acetone as solvent. The compounds were synthesized in good yields (53–85%). Low cost, high yields and easy availability of compounds helped in the synthesis. Electron withdrawing groups favor the formation of stable phenoxide ion thus in turn favors the formation of product while electron donating groups do not favor the reaction. Phenol derivatives gave good yields as compared to that of aniline. As aprotic polar solvents favor SN2 type reactions so acetone provided best solvation for the reactions. K2CO3 was proved to be good for the synthesis. Antibacterial, Antiurease and NO scavenging activity of synthesized compounds were also examined. 4-bromo-2-chloro-1-(prop-2-ynyloxy)benzene 2a was found most active compound against urease enzyme with a percentage inhibition of 82.00±0.09 at 100 µg/mL with IC50 value of 60.2. 2-bromo-4-methyl-1-(prop-2-ynyloxy)benzene 2d was found potent antibacterial against Bacillus subtillus showing excellent inhibitory action with percentage inhibition of 55.67±0.26 at 100 µg/ml wih IC50 value of 79.9. Based on results, it can be concluded that some of the synthesized compounds may have potential antiurease and antibacterial effects against several harmful substances.

Introduction

(Prop-2-ynyloxy)benzene is an important compound having vast synthetic applications. Most important of all of its applications is its use as triazole intermediate [1]. Synthetic methods of organic chemistry reveal alkyne groups to be most reactive ones which increase the importance of the compound [2]. Low cost, high yield of the compound has attracted the attention of scientists to synthesize the compound as monomer for many polymeric compounds as it is thermally reactive end capping agent. [3] The reaction became much more important due to presence of terminal alkyne which helps in easy polymerization reactions under mild reaction condition, low temperature which is more favorable for linear and graft polymeric compounds. [4] (Prop-2-ynyloxy)benzene structure is mainly found in compounds of medicinal importance specially of antimicrobial importance. [5] (Prop-2-ynyloxy)benzene type compounds are also used in synthesis of large number of polymeric compounds mainly included lignin [6], azide functionalized dendrons [7], and β lactam fused enediynes [8], mono and bis-1,2,3-triazole acyclonucleoside [9], symmetrical and asymmetrical, cyclic and acylic enediynes [10] and sulfonated polytriazole proton exchange membrane for methanol feul cells via click chemistry. [4] Synthetic importance of the compound is revealed from its structure having easy approach for upcoming substances joining them to form complex molecules more accurately with great ease [11].

Haung and coworkers observed that (Prop-2-ynyloxy)benzene type compounds showed antimicrobial, antitumor, anti HIV, DNA scavenging inhibitors, anti proliferating, anti obesity and central nervous system activities. [4] The main intention of the present work is the synthesis of (Prop-2-ynyloxy)benzene derivatives by reported methodologies [12][20] and our focused to adopt optimization reaction condition such as effect of solvents, moler concentration of bases, stirring and refluxing time. In addition evaluating their antibacterial, antiurease and NO scavenging activities.

Results and Discussion

Chemistry

Krim and co-workers reported that (Prop-2-ynyloxy)benzene is an intermediate in the synthesis of triazole via Huigen dipolar cycloaddition method using click chemistry. Terminal alkynes provide an efficient method for the synthesis of triazole, [21] which can be prepared by reaction of 3-Bromopropyne and aniline derivatives in DMF [9], [22]. We reported the reaction of phenol and propargyl bromide for the synthesis of (Prop-2-ynyloxy)benzene derivatives under different reaction conditions. [23][28] There is very little work on the synthesis of reported compounds under these optimized reaction conditions and so far no information was published for antibacterial, antiurease and NO scavenging activity of the synthesized compounds.

We optimized the reaction conditions like temperature, solvents, bases and its concentration for the synthesis of compounds (2a–2f) (Fig. 1, Table 1). It was observed that successive addition of various bases and solvent types helped to obtain moderate to very good yields (74–85%) of synthesized compounds. For the synthesis of compound (2d) 2-bromo-4-methyl-1-(prop-2-ynyloxy) benzene, 2-bromo-4-methyl phenol (0.535 mmol) was reacted with propargyl bromide (1.2 eq.) in THF using LiH (2 eq.) as base at room temperature producing no product. It was noted that 2d was synthesized with almost 74% yield, when the reaction was carried out in acetone and K2CO3 (2 eq.) under reflux conditions for 5 hrs. Compound, 1-nitro-4-(prop-2-ynyloxy) benzene (2f) was prepared by using dry acetone and K2CO3 (3.5 eq.) under reflux conditions for 5 hour producing 76% yield. However when compound was first stirred in the presence of K2CO3 (3.5 eq.) and acetone at room temperature for 2 hour then adding propargyl bromide (1.3 eq.) to the reaction mixture and stirring for 16 hour resulted in 53% yield. It was surprisingly noted that acidity of phenoxide ion was also affected by nature of substitutions. Electron withdrawing groups might increase the acidity to led enhanced yield of products while electron donating groups allowed to decrease the yield of synthesized compounds (Fig. 1, Table 1).

Compound 4a and 4b were prepared by reaction of 4,4′-(propane-2,2-diyl)diphenol (0.87 mmol) and 4,4′-sulfonyldiphenol (0.8 mmol) with propargyl bromide (2.4 eq.) in acetone using K2CO3 (4 eq.) as base under reflux conditions (80°C) respectively. (Fig. 2, Table 2). It was noteworthy that 4b gave very good yield (73%) due to more acidic protons of 4,4′-sulfonylbis((prop-2-ynyloxy)benzene) while 4a gave moderate yield (70%) of 4,4′-(propane-2,2-diyl)bis((prop-2-ynyloxy)benzene) due to less acidic protons because of two methyl moieties present in the molecule.

It was observed that naphthalene-1-ol (0.694 mmol) were allowed to react with propargyl bromide (1.2 eq.) in the presence of K2CO3 base (3.5 eq.) and 3 ml dry acetone as solvent to synthesize 1-(prop-2-ynyloxy)naphthalene 6a, (71%). Naphthalene-2-ol (0.694 mmol) was first stirred in the presence of K2CO3 (3.5 eq.) and acetone at room temperature for 2 hour then adding propargyl bromide (1.3 eq.) to the reaction mixture and stirring for 16 hour resulted in 73% yield of 2-(prop-2-ynyloxy)naphthalene 6b. The reaction completion was monitored by TLC. No product was formed while refluxing the reaction mixture with 1 eq. of K2CO3 base. It was surprisingly noted that increasing the eq. of K2CO3 base led to reaction completion (Fig. 3, Table 3).

Moreover substituted aniline 7 (2.68 mmol) and 7a (0.699 mmol) were reacted with propargyl bromide (1.3 eq.) to synthesize variously substiuted (prop-2-ynyl)anilines, 8a–8d (Fig. 4, Table 4). It was noteworthy that reaction time was prolonged as compared to phenolic compounds to provide more harsh conditions in order to obtain desired products. In the presence of 1 eq. K2CO3 base 7a was allowed to react with propargyl bromide (1.3 eq.), it was surprisingly noted that reaction was failed however using 2–3 eq. base proved helpful for the completion of reaction with very good yield of 8d (69%).

We explained a similar application of present method to obtain 10, N-(prop-2-ynyl)-4-(prop-2-ynyloxy)aniline (Fig. 5, Table 5). It was observed that reaction was carried out at room temperature by 16 hour stirring of 4-aminophenol (0.917 mmol) and K2CO3 (2 eq.) as base with addition of propargyl bromide (2.4 eq.) in DMF as solvent during time interval of 20 min, after this reaction mixture was refluxed for 48 h under nitrogen atmosphere to obtain desired product 10, no product was formed. It was noteworthy that when the reaction was done with K2CO3 (4 eq.) as base in dry acetone as solvent, compound 10 was obtained with 76% yield.

Reaction of 11 4-amino-5-hydroxynaphthalene-2,7-disulfonic acid (0.313 mmol) and 11a 4-amino-3-hydroxynaphthalene-1-sulfonic acid (0.418 mmol) with propargyl bromide (2.7 eq.) in basic conditions did not confirm the formation of any product. It was noted that the reaction failed even after addition of excess quantities of base and refluxing the reaction for prolonged time (Fig. 6). It might be possible that failure of reactions were due to presence of sulfonic acid functionality present in the molecule.

Pharmacology

Antiurease activity.

Electron donating groups enhance the activity of compounds by donating the electrons to phenyl group which enhances the binding of enzyme active site and the molecule easily [29]. Bromine containing compounds more easily inhibit the urease activity rather than compounds containing fluorine and chlorine due to lesser electronegativity values of bromine. Methyl substituent and sulfur moiety also increases the activity of compounds [30]. The compounds were tested to inhibit the activity of urease enzyme at 50 µg/ml and 100 µg/ml concentrations and their results are given in table 3. Thiourea was used as positive control with percentage inhibition activity of 65±0.01 with IC50 value of 23.3 at 40 µg/ml.

The results revealed that electronic effects of aryl substitutions are playing a significant role in activity manipulation. 2a, 2b and 2e contain bromine substitution which enhances their activity due to lesser withdrawing effect. Chloro and flouro groups decrease the activity of compounds due to more withdrawing effect. 4a also exhibited remarkable results due to presence of methyl moieties. 2f surprisingly gave contrasting results. As nitro group has withdrawing electronic effect, which decreases the inhibitory effect of synthesized compound. But results showed comparatively good results for the respective compound.

It was observed that all synthesized compounds 2a-10 showed moderate to very good urease inhibition activity. 4-bromo-2-chloro-1-(prop-2-ynyloxy)benzene 2a showed comparatively good results with a percentage inhibition of 42.5±0.7 at 50 and 82.00±0.09 at 100 µg/mL with IC50 value of 60.2. The 2b–6a showed good inhibition activities with percentage inhibition of 81.07±0.1, 86.45±0.07, 85.89±0.07, 79.59±0.11, 65.8±0.18, 89.42±0.05, 74.9±0.13, 65.3±0.18 at 100 µg/ml with IC50 values ranging between 64.92, 70.1, 67.14, 62.15, 63.7, 62.3, 70.9 and 68 µg/ml respectively (Table 6). 6b did not show any satisfactory results.

Aniline derivatives showed comparatively lesser activities due to lack of binding site at tertiary amine site. Ni acts as cofactor for urease enzyme, which helps in chelation of ions by compounds containing amino groups, thus activating the enzyme activity but in tertiary amines, chelation do not occur easily. Compound 8a-10 (Table 6) showed low inhibition activities with inhibition values of 55.65±0.239, 50.83±0.265, 56.5±0.234, 51.5±0.26 at 100 µg/ml with IC50 values of 90.7, 98.2, 89.6 and 91.7 µg/ml respectively. Compound 8c did not show any satisfactory results.

Nitric oxide (NO) scavenging activity

Nitric oxide is a free radical causing oxidative damage to biomolecules, finally leading to many disorders like cancer [31], aging and diabetes [32]. The synthesized compounds (2a-10) exhibited good nitric oxide scavenging activity. Vitamin C (Ascorbic acid) was used as a control with percentage inhibition values 38.5±0.16 at 50 and 84.1±0.12 at 100 µg/mL with IC50 value 50.43.

Electron withdrawing groups decreases the activity while electron donating groups enhances the nitric oxide scavenging activity. Bromine containing compounds exhibited more activity as 2c has maximum inhibitory activity due to the presence of two bromine groups attached to aromatic ring. 4a also exhibited remarkable results due to the presence of methyl moieties having donating effects thus enhancing the activity. 2a, 2b and 2e have relatively lower IC50 values due to electron withdrawing effects of fluorine and chlorine groups. 4b having SO2 groups showed lesser inhibitory activity. Aniline derivatives also showed lower inhibitory activity as compared to that of phenolic ones.

Compounds 2,4-dibromo-1-(prop-2-ynyloxy)benzene 2c and 4,4′-(propane-2,2-diyl)bis((prop-2-ynyloxy)benzene) 4a were found good percentage inhibitors against NO free radicals with value 39.67±0.09 at 50 µg/mL and 77.03±0.11 at 100 µg/mL with IC50 value 63.8 and percentage inhibition 40±0.09 at 50 µg/mL and 74.53±0.13 at 100 µg/mL with IC50 value 64.4 respectively. 2a, 2b and 2e exhibited fairly good NO percentage inhibition (76.09±0.12, 68.44±0.16 and 77.5±0.11 at 100 µg/mL) with IC50 values 67.72, 67.9 and 68.8 respectively (Table 7). Compounds 2d, 4b-10 were comparatively less active to scavenge NO free radicals (69.95±0.15, 59±0.15, 63.17±0.19, 65.19±0.18, 68.65±0.16, 57.98±0.21, 62.8±0.19 and 65.5±0.18 at 100 µg/mL) with IC50 values 89.6, 83, 76.16, 79.4, 87.5, 78 and 78.1 for each compound respectively. 1-nitro-4-(prop-2-ynyloxy)benzene 2f and N-(prop-2-ynyl)naphthalen-1-amine 8d did not show any significant results and aniline derivatives showed lesser activity than that of phenol.

Antibacterial activity

Disc diffusion method was adopted to test the antibacterial activity of synthesized compounds 2a-10 against four different bacterial strains. Ampiciline was used as positive control to check antibacterial activity of synthesized compounds. The bacterial strains employed were: Escherichia coli, Bacillus subtillus, Shigella flexneri, Staphylococcus aureus, Pseudomonas aeroginosa and Salmonella typhi. Antibacterial activity of synthesized compounds 2a–2e, 4a, 4b and 10 showed moderate activity against Staphylococcus aureus while 2f, 6a and 8a–8d were found to be inactive (Table 8).

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Table 8. Antibacterial activity of (prop-2-ynyloxy)benzene derivatives.

https://doi.org/10.1371/journal.pone.0115457.t008

Hence compounds 2c, 2f, 4b–8a and 10 have batter activity against Bacillus subtillus. It was noted that 2d was found potent antibacterial against Bacillus subtillus showing good inhibitory action. 2a, 2b, 2e, 4a, and 8b–8d did not show any satisfactory result aginst Bacillus subtillus and were found to be inactive (Table 8). All of the synthesized compounds 2a-10 were proved inactive against Escherichia coli, Shigella flexneri, Pseudomonas aeroginosa and Salmonella typhi providing no significant results.

Materials and Methods

Ethics statement

No specific permits were required for these field studies. No specific permissions were required for these locations/activities because they were not carried out on privately owned or protected areas. The field studies did not involve endangered or protected species.

General

Melting points were determined using a Buchi melting point B-540 apparatus. All reagents used were purchased from Alfa Aesar, Sigma Aldrich and E. Merck Company. 13C-NMR and 1 H-NMR spectra were recorded on Bruker ARX 400 MHz FT-NMR spectrophotometer and Bruker ARX 600 MHz FT-NMR spectrophotometer while relishing CDCl3 as internal reference for the recording of spectra. The chemical shift was given in δ in ppm and coupling constant in Hz. EI-MS spectras were recorded on a JMS-HX-110 spectrometer, with a data system. For column chromatography technique, silica gel (70–230 mesh) and silica gel (230–400 mesh) were used. The reactions were monitored on TLC, using Merk Silica gel 60PF254 cards. The compounds were visualized by UV lamp (254, 365).

General procedure for the synthesis of (Prop-2-ynyloxy)benzene derivatives

The synthesis of (Prop-2-ynyloxy)benzene derivatives was carried out by using four methods.

Method A

To 1 eq (0.1 g) phenol (Figs. 16) added (1.2–2.7 eq.) propargyl bromide and (2–4 eq.) K2CO3 in acetone (3 ml). The reaction mixture was refluxed at 80°C till reaction completion based on TLC monitoring. On completion the reaction mixture was cooled up to room temperature and solvent was removed under reduced pressure. The residue was dissolved in distilled water and then extraction was done by twice addition of methylene chloride. Organic layer was separated and dried with the help of anhydrous magnesium sulphate. Solvent was removed under reduced pressure to obtain the desired product. After evaporation and filtration process the product was purified by flash chromatography. 60/40 mixture of hexane and ethyl acetate was used as eluent [5], [33][34].

Method B

1 eq (0.1 g) of phenol (Fig. 1) and (1.2 eq.) of LiH was stirred in 6 ml of THF at room temperature for 30 min, added (2 eq.) propargyl bromide to the reaction mixture and stirred overnight at room temperature. Reaction completion was monitored by TLC conditions. Evaporated the solvent under reduced pressure to obtain the final product. Product was purified by flash chromatography. 60/40 mixture of hexane and ethyl acetate was used as eluent [9], [35].

Method C

Phenol (Fig. 1 and 3) 1eq (0.1 g) and K2CO3 (3.5 eq.) was stirred at room temperature for two hours in 0.6 ml acetone. Propargyl bromide (1.3 eq.) in 1.66 ml acetone was added in 20 min and refluxed the reaction mixture for 16 hours under nitrogen atmosphere. Reaction completion was monitored by TLC. After the reaction completion solvent was removed under reduced pressure. The residue was dissolved in distilled water and organic layer was extracted using dichloromethane thrice. Additional water was removed by drying with anhydrous MgSO4 [11], [36].

Method D

1 eq (0.1 g) of 4-aminophenol (Fig. 5) and (3.5 eq.) anhydrous potassium carbonate in 0.6 ml dry DMF was stirred at room temperature for about 16 hours. 2.4 eq propargyl bromide in 0.48 ml dry DMF was added slowly in about 15 min. the reaction mixture was heated at 80°C for 48 hours under nitrogen atmosphere. Reflux conditions were used for heating the reaction mixture. Reaction completion was monitored by TLC and reaction time varied according to the condition of product. After reaction completion the solvent was removed reduced pressure. Residue was dissolved in water and then organic compound was extracted by addition of dichloromethane thrice the volume of water. Excess water was removed by adding MgSO4. The solvent was removed under reduced pressure and the compound separated was recrystallised using methanol thrice [11], [36].

Synthesis of 4-bromo-2-chloro-1-(prop-2-ynyloxy)benzene (2a).

Brown solid, M.p. 29°C, 79% Yield, 1H-NMR (400 MHz, CDCl3): δ = 7.50 (d, J = 2.4 Hz, 1H-Ar), 7.33–7.25 (m, 1H-Ar), 6.96 (d, J = 8.8 Hz, 1H-Ar), 4.74 (d, J = 2.4 Hz, 2H-CH2), 2.53 (t, 1H-acetylene). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 156.5, 134, 129.6, 122.2, 75.2, 51.2; EIMS (m/z -ion mode): 245 [M-H]; [M-CH2CCH fragment] = 206; [M-Br fragment] = 165.

Synthesis of 2-bromo-4-chloro-1-(prop-2-ynyloxy)benzene (2b).

Brown liquid. 78% Yield, 1H-NMR (400 MHz, CDCl3): δ = 7.54 (d, J = 2.4 Hz, 1H-Ar), 7.23–7.15 (m, 1H-Ar), 6.98 (d, J = 8.8 Hz, 1H-Ar), 4.77 (d, J = 2.0 Hz, 2H-CH2), 2.53 (t, 1H-acetylene). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 161, 137.5, 131, 125.2, 115.5, 76.8, 72.1, 51.2; EIMS (m/z -ion mode): 245 [M-H]; [M-CH2CCH fragment] = 206; [M-OCH2CCH fragment] = 191; [M-Br fragment] = 165.

Synthesis of 2,4-dibromo-1-(prop-2-ynyloxy)benzene (2c).

Brown solid. M.p. 157.95°C, 85% Yield, 1H-NMR (600 MHz, CDCl3): δ = 7.68 (d, J = 2.4 Hz, 1H-Ar), 7.39–7.30 (m, 1H-Ar), 6.95 (d, J = 9.0 Hz, 1H-Ar), 4.76 (d, J = 2.4 Hz, 2H-CH2), 2.55 (t, 1H-acetylene). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 167.9, 140.7, 127.8, 116, 76.5, 74.1, 51.2; EIMS (m/z -ion mode): 289 [M-H]; [M-CH2CCH and CO fragment] = 221; [M-Br fragment] = 207; [M-OCH2CCH and Br fragment] = 149.

Synthesis of 2-bromo-4-methyl-1-(prop-2-ynyloxy)benzene (2d).

Yellow oily liquid. 74% Yield, 1H-NMR (400 MHz, CDCl3): δ = 7.27 (d, J = 2.4 Hz, 1H-Ar), 6.97–6.88 (m, 1H-Ar), 6.67 (d, J = 9 Hz 1H-Ar), 4.69 (d, J = 2.1 Hz, 2H-CH2), 2.99 (t, 1H-acetylene), 1.55 (s, 3H-CH3). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 157, 138.9, 129, 79.3, 75.7, 51.2, 19.8; EIMS (m/z -ion mode): 224 [M-H]; [M-CH2CCH fragment] = 186; [M-Br fragment] = 145; [M-CH2CCH and Br fragment] = 107.

2-bromo-4-fluoro-1-(prop-2-ynyloxy)benzene (2e).

Brown liquid. 79% Yield, 1H-NMR (400 MHz, CDCl3): δ = 7.02–6.97 (m, 3H-Ar), 4.72 (d, J = 2. 4 Hz, 2H-CH2), 2.51 (t, 1H-acetylene). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 154.8, 124.3, 120.1, 79.4, 55.6; EIMS (m/z -ion mode): 231 [M-H]; [M-CH2CCH fragment] = 193; [M-CO and CH2CCH fragment] = 165.

Synthesis of 1-nitro-4-(prop-2-ynyloxy)benzene (2f).

Yellow crystals. M.p. 129°C, 76% Yield, ­1H-NMR (400 MHz, CDCl3): δ = 8.216 (d, J = 9 Hz, 1H-Ar), 7.05 (d, J = 9 Hz, 1H-Ar), 4.77 (d, J = 2.4 Hz, 2H-CH2), 2.56 (t, 1H-acetylene). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 166.5, 142.2, 129, 77, 57.8; EIMS (m/z -ion mode): 177 (M-H]; [M-NO2 fragment] = 131; [M-NO2 and OCH2CCH fragment] = 77.

Synthesis of 4,4′-(propane-2,2-diyl)bis((prop-2-ynyloxy)benzene) (4a).

Light brown greasy liquid. 73% Yield, 1HNMR (600 MHz, CDCl3): δ = 7.164–7.147 (m, 4H-Ar), 7.103–7.083 (m, 4H-Ar), 4.66 (d, J = 2.4 Hz, 4H-CH2), 2.90 (s, 6H-CH3), 2.52 (t, 2H-acetylene). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 156, 151, 130.9, 117, 77.5, 31.8; EIMS (m/z -ion mode): 304 [M-H]; [M-CH3 fragment] = 289; [M-CH2CCH fragment] = 266; [M-benzene, CH3 and OCH2CCH fragment] = 169.

Synthesis of 4,4′-sulfonylbis((prop-2-ynyloxy)benzene) (4b).

White solid. M.p. 136°C, 69% Yield, 1H-NMR (400 MHz, CDCl3): δ = 7.86–7.83 (m, 4H-Ar), 7.24–7.00 (m, 4H-Ar), 4.71 (d, J = 2. 4 Hz, 4H-CH2), 2.52 (t, 2H-acetylene). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 170, 140.7, 128, 119.3, 80.4, 77.5; EIMS (m/z -ion mode): 325 [M-H]; [M-CH2CCH fragment] = 288; [M-2CH2CCH fragment] = 250; [M- CH2CCH and CO fragment] = 223; [M- CH2CCH and 2CO fragment] = 195.

Synthesis of 1-(prop-2-ynyloxy)naphthalene (6a).

Light brown liquid. 71% Yield, 1H-NMR (400 MHz, CDCl3): δ = 7.77–7.73 (m, 3H-Ar), 7.45–7.32 (m, 3H-Ar), 7.18–7.09 (m, 1H-Ar), 4.80 (d, J = 2.0 Hz, 2H-CH2), 2.53 (t, 1H-acetylene). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 158, 135, 130.6, 126, 77.8, 63; EIMS (m/z -ion mode): 182 [M-H]; [M-CH2CCH fragment] = 143; [M-CH2CCH and CO fragment] = 115; [M-benzene and OCH2CCH fragment] = 77.

Synthesis of 2-(prop-2-ynyloxy)naphthalene (6b).

Dark brown greasy liquid. 73% yield, 1H-NMR (400 MHz, CDCl3): δ = 7.82–6.75 (m, 7H-Ar), 4.81 (d, J = 2.07 Hz, 2H-CH2), 3.01 (t, 1H-acetylene). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 160, 128.3, 126, 122.7, 116.5, 77.5, 58; EIMS (m/z -ion mode): 182 [M-H]; [M-CH2CCH fragment] = 143; [M-CH2CCH and CO fragment] = 115; [M-benzene and OCH2CCH fragment] = 77.

Synthesis of 4-bromo-2-methyl-N,N-di(prop-2-ynyl)aniline (8a).

Dark brown liquid. 68% Yield, 1H-NMR (400 MHz, CDCl3): δ = 7.20 (d, J = 1.96 Hz, 1H-Ar), 7.14–7.05 (m, 1H-Ar), 6.58 (d, J = 9 Hz, 1H-Ar), 3.94 (d, J = 2.4 Hz, 2H-CH2), 2.71 (t, 1H-acetylene), 2.18 (s, 3H-CH3), 1.18 (s, 1H-NH). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 154.2, 140, 138.9, 120.4, 79, 75.3, 45, 18.5; EIMS (m/z -ion mode): 224 [M-H]; [M-CH2CCH fragment] = 184; [M-Br fragment] = 144; [M-CH2CCH and Br fragment] = 106.

Synthesis of 2-bromo-4-methyl-N-(prop-2-ynyl)aniline (8b).

Dark brown liquid. 71% Yield, 1H-NMR (400 MHz, CDCl3): δ = 7.26 (s, 1H-Ar), 6.98–6.91 (m, 1H-Ar), 6.64 (d, J = 8 Hz, 1H-Ar), 3.96–3.90 (m, 4H-CH2), 2.22 (t, 2H-acetylene), 1.55 (s, 3H-CH3). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 140.5, 137.8, 129.9, 117, 80, 76.5, 46, 20.9; EIMS (m/z -ion mode): 224 [M-H]; [M-CH2CCH fragment] = 184; [M-Br fragment] = 144; [M-CH2CCH and Br fragment] = 106.

Synthesis of 4-bromo-3-methyl-N-(prop-2-ynyl)aniline (8c).

Dark brown solid. M.p. 40.85°C, 70% Yield, 1H-NMR (400 MHz, CDCl3): δ = 7.17 (d, J = 6 Hz, 1H-Ar), 6.63–6.55 (m, 1H-Ar), 6.36 (d, J = 7 Hz, 1H-Ar), 4.06 (d, J = 2.4 Hz, 4H-CH2), 2.86 (t, 2H-acetylene), 2.18 (s, 3H-CH3). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 150.6, 135.9, 130, 117.2, 80, 75, 45, 23.8; EIMS (m/z -ion mode): 224 [M-H]; [M-CH2CCH fragment] = 184; [M-Br fragment] = 144; [M-CH2CCH and Br fragment] = 106.

Synthesis of N-(prop-2-ynyl)naphthalen-1-amine (8d).

Brown greasy liquid. 69% Yield, 1H-NMR (400 MHz, CDCl3): δ = 7.78–6.7 (m, 7H-Ar), 4.11 (d, J = 2.40 Hz, 2H-CH2), 2.88 (t, 1H-acetylene), 2.26 (s, 1H-NH). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 149, 135, 126, 107.6, 80.8, 75.5, 31; EIMS (m/z -ion mode): 180 [M-H]; M-CH2CCH fragment] = 143; [M-HCN fragment] = 115; [M-benzene and HNCH2CCH] = 77.

Synthesis of N-(prop-2-ynyl)-4-(prop-2-ynyloxy)aniline (10).

Dark brown greasy liquid. 53% Yield, 1H-NMR (400 MHz, CDCl3): δ = ): δ = 7.24–7.03 (m, 4H-Ar), 3.98 (d, J = 2.40 Hz, 6H-CH2), 2.88 (t, 3H-acetylene). 13C-NMR (100 MHz, CDCl3+ CD3OD): δ = 149, 145.5, 118.7, 118.6, 76, 71, 61, 45; EIMS (m/z -ion mode): 185 = [M-H]; [M-CH2CCH fragment] = 146; [M-2CH2CCH fragment] = 108; [M-2CH2CCH and CO fragment] = 80.

Antiurease Activity

Antiurease activity of the synthesized compounds was determined by using the modified phenol-hypochlorite method [37]. Varying concentrations were used to prepare the urease inhibition curve of Thiourea (a known inhibitor of Urease) in the reaction mixture. Phosphate buffer solution of pH 7 containing 1 unit of enzyme was added about 200 µl in the test tube which was followed by the addition of 230 µl of phosphate buffer and 20 µl of thiourea from a scrupulous stock solution for the particular concentration of thiourea in the reaction mixture respectively. Test sample solutions were added instead of thiourea solution in case of test samples. The reaction mixture were mixed and incubated at 25°C for 5 min. 400 µl of urea stock (20 mM) was added in each test tube. In order to allow the action of urease on urea, all the test tubes were incubated at 40°C.

Phenol hypochlorite reagent (1150 µl) was added after 10 min incubation at 40°C. Test tubes were again incubated at 56°C for 25 min. A blue coloured complex was formed in the reaction mixture. The samples were allowed to cool for 5 min and then their absorbance values were measured at 625 nm and the %age inhibition was calculated. Experiment was repeated three times to get the SD values.

NO Scavenging Activity

Solutions of known concentration were prepared by dissolving the synthesized compounds in appropriate solvents. [38] A mixture of sample solution (20 µl), 10 mM sodium nitropruside solution (200 µl) and phosphate buffer of 7.4 pH (1780 µl) was taken in a dry test tube and the mixture was incubated at 37°C for 2 hours. 200 µl Griess reagent (0.3% Sulphanilic acid in glacial acetic acid, 0.1% Naphthyl ethylene diamine HCL (NED): Mixed equal volume of both solutions just before use) was added after the incubation period is over. After 20 min keeping the solution at room temperature, its absorbance at 528 nm was measured. Each sample was analyzed in triplicate. Positive control of any known antioxidant and control of all samples were run in parallel. Use of negative control was to calibrate the instrument. Sample blanks were also prepared in case if the samples were coloured. Correction in the absorbance values was made by subtracting the sample blank reading from the sample reading.

Antibacterial Activity

Disc diffusion method was used for the determination of antibacterial activity. [39] synthetic organic compounds were examined to evaluate their antibacterial potential against four different strains of bacteria i.e. Staphlococcus aureus (Gram + ive), Pseudomonas- aeroginosa (Gram−ive) and Escherichia coli (Gram−ive), Bacillus subtilis (Gram + ive), Salmonella typhi (Gram−ive) and Shigella flexneri (−ive). The synthetic organic compounds were dissolved in an appropriate solvent to get the solution of known concentration (5 µg/µl or 10 µg/µl). In order to prepare discs of different potencies known volume of extract/synthetic organic compound solution was applied on the discs, air dried and used. After preparation of the sterilized nutrient agar and nutrient broth medium, the sterilized nutrient agar medium was poured under aseptic conditions into sterile glass petri dishes/plates. A known volume of bacterial growth culture can also be poured on the surface of agar medium solidified in petri dishes. After inoculation petri plates were kept at room temperature for 15–30 minutes so that the poured medium is adsorbed. All the discs, including test sample, +ive and –ve controls were placed onto the inoculated nutrient agar medium solidified in petri dishes with the help of sterile forceps and incubated the petri plates at 37°C±2°C for 16–24 hours. Zone of inhibition was measured in mm using ordinary scale. Experiment was repeated three times to get the SD values.

Conclusions

We have reported the synthesis of (prop-2-ynyloxy)benzene derivatives with efficient yields under mild conditions. Some of the reactions were proceeded using comparatively harsh conditions. Using more than one method helped in comparison of reaction conditions and finding out the best one. Potassium carbonate is a mild base, which provides best reaction conditions. Other bases used were comparatively harsh which may also affect the aromatic ring rendring it unfavorable for reaction products. Harsh bases like LiH, NaOH deactivate the aromatic ring. Acetone favours SN2 type reaction providing aprotic polar conditions so acetone and potassium carbonate were proven to be best for the synthesis of desired products. The synthesized compounds were proved to be good antiurease, antioxidant and antibacterial substances. Phenolic Compounds containing electron donating groups were proved to be better urease inhibitors and good NO scavenging activity than those compounds containing electron withdrawing groups. In contrast synthesized phenolic compounds containing both electron donating and electron withdrawing functionality gave better results against bacterial strains. It also noted Aniline derivatives showed lesser activity than phenolic compounds. The results of this study revealed that these compounds may have potential inhibitory activity and can be used as antibacterial, antiurease and antioxidant molecules.

Acknowledgments

The present data is the part of M.Phil thesis research work of Tannaza Batool. The authors gratefully acknowledge the financial support by HEC (HEC Project No. 20-1465/R&D/09/5458).S. U. Khan would like to extend his sincere appreciation to the Deanship of Scientific Research at King Saud University for its funding through the Research Group Project no RGP-VPP-255.

Author Contributions

Conceived and designed the experiments: NR FN MZ. Performed the experiments: TB YG MN AY. Analyzed the data: UAR SUK MZH. Contributed to the writing of the manuscript: MZH HZEJ.

References

  1. 1. Meldal M, Tornoe CW (2008) Cu-Catalyzed Azide-Alkyne Cycloaddition. Chemical Reviews 108:2952–3015.
  2. 2. Antoni P, Nystro D, Hawker CJ, Malkoch M (2007) A chemoselective approach for the accelerated synthesis of well-defined dendritic architectures. Chemical Communications 22:2249–2251.
  3. 3. Wei L, Mulder JA, Xiong H, Zi CA, Douglas CJ, et al. (2001) Efficient preparations of novel ynamides and allenamides. Tetrahedron 57:459–466.
  4. 4. Huang YJ, Ye YS, Yen YC, Tsai LD, Hwang BJ, et al. (2011) Synthesis and characterization of new sulfonated polytriazole proton exchange membrane by click reaction for direct methanol fuel cells (DMFCs). International Journal of Hydrogen Energy 36:15333–15343.
  5. 5. He J, Feng L, Li J, Tao R, Wang F, et al. (2012) Design, synthesis and biological evaluation of novel 2-methylpyrimidine-4-ylamine derivatives as inhibitors of escherichia coli pyruvate dehydrogenase complex E1. Bioorganic & Medicinal Chemistry 20:1665–1670.
  6. 6. Zhang Y, Wang G, Huang J (2011) Preparation of amphiphilic poly(ethylene oxide)-block-polystyrene macrocycles via glaser coupling reaction under cubr/pyridine system. Journal of Polymer Science Part A: Polymer Chemistry 49:4766–4770.
  7. 7. Lee JW, Kim JH, Kim B-K, Kim JH, Shin WS, et al. (2006) Convergent synthesis of PAMAM-like dendrimers from azide-functionalized PAMAM dendrons. Bulletin of the Korean Chemical Society 27:1795–1800.
  8. 8. Basak A, Pal R, Das S (2009) Synthesis of -lactam fused enediynes by intramolecular kinugasa reaction: comparison of reactivity with monocyclic analogues. International Journal of Chemistry 1:63–74.
  9. 9. Krim J, Sillahi B, Taourirte M, Rakib EM, Engels JW (2009) Microwave-assisted click chemistry: synthesis of mono and bis-1,2,3-triazole acyclonucleoside analogues of acyclovir via copper (i)-catalyzed cycloaddition. Arkivoc 8:142–152.
  10. 10. Joshi MC, Bisht GS, Rawat DS (2007) Syntheses and antibacterial activity of phendioxy substituted cyclic enediynes. Bioorganic & Medicinal Chemistry Letters 17:3226–3230.
  11. 11. Kundu NG, Chaudhuri G, Upadhyay A (2001) Palladium-catalyzed heteroannulation leading to heterocyclic structures with two heteroatoms: a highly regio- and stereoselective synthesis of (Z)-4-alkyl-2-alkyl(aryl)idene-3,4-dihydro-2 H-1,4-benzoxazines and (Z)-3-alkyl(aryl)idene-4-tosyl-3,4-dihydro-2 H. The Journal of Organic Chemistry 66:20–29.
  12. 12. Booth SE, Jenkins PR, Swain CJ, Sweeney JB (1994) Intramolecular addition of vinyl and aryl radicals to oxime ethers in the synthesis of five-, six- and seven-membered ring systems. Journal of the Chemical Society, Perkin Transactions 1 34:99–102.
  13. 13. Samai S, Nadi GN, Singh MS (2010) An efficient and facile one pot synthesis of propargylamines by three-component coupling of aldehydes, amines and alkynes via C-H activation catalyzed by NiCl2.Tetrahedron Letters. 51:5555–5558.
  14. 14. Rostovstev VV, Green LG, Fokin VV, Sharpless KB (2002) A stepwise Huisgen cycloaddition process: Copper (I) catalyzed regioselective ligation of azides and terminal alkynes. Angewandte Chemie International Edition 41:2596–2599.
  15. 15. Tornoe C, Christensen C, Medal M (2002) Peptidotriazoles on solid phase: [1,2,3]-triazoles by regiospecific copper(I)-catalyzed 1,3-dipolar cycloadditions of terminal alkynes to azides. Journal of Organic Chemistry 67:3057–3064.
  16. 16. Agalave SG, Maujan SR, Pore VS (2011) Click chemistry: 1,2,3-Triazoles as pharmacophores. Chemistry-An Asian Journal 6:2696–2718.
  17. 17. Speers AE, Adam GC, Cravatt BF (2003) Activity-based protein profiling in vivo using a copper (I)-catalyzed azide-alkyne [3+2] cycloaddition. Journal of American Chemical Society 125:4686–4687.
  18. 18. Dabiri M, Salehi P, Bahramnejad M, Sherafat F (2010) Synthesis of diheterocyclic compounds based on triazolyl methoxy Phenylquinazolines via a one-pot four-component-click reaction. Journal of Combinatorial Chemistry 12:638–642.
  19. 19. Quan ZJ, Quion X, Zhang Z, Da YX, Wang XC (2013) Copper-catalyzed click synthesis of functionalized 1,2,3-triazoles with 3,4-dihydropyrimidinone or amide group via a one-pot four-component reaction. Tetrahedron 69:881–887.
  20. 20. Martın-Matute B, Nevado C, Cárdenas DJ, Echavarren AM (2003) Intramolecular Reactions of Alkynes with Furans and Electron Rich Arenes Catalyzed by PtCl2: The Role of Platinum Carbenes as Intermediates. Journal of American Chemical Society 125:5757–5766.
  21. 21. Agag T, Takeichi T (2001) Novel Benzoxazine Monomers Containing p-Phenyl Propargyl Ether: Polymerization of Monomers and Properties of Polybenzoxazines. Macromolecules 34:7257–7263.
  22. 22. Pal M, Parasuraman K, Yeleswarapu KR (2003) Palladium-catalyzed cleavage of O/N-Propargyl protecting groups in aqueous media under a copper-free condition. Organic Letters 5:349–352.
  23. 23. Wang Q, Chan TR, Hilgraf R, Fokin VV, Sharpless KB, et al. (2003) Biconjugation by Copper(I)-catalyzed azide-alkyne [3+2] cycloaddition. Journal of American Chemical Society 125:3192–3193.
  24. 24. Bahulayan D, Arun S (2012) An easy two step synthesis of macrocyclic peptidotriazoles via a four-component reaction and catalyzed intramolecular alkyne-azide [3+2] click cycloaddition. Tetrahedron Letters 53:2850–2855.
  25. 25. Quan ZJ, Quion X, Zhang Z, Da YX, Wang XC (2013) Copper-catalyzed click synthesis of functionalized 1,2,3-triazoles with 3,4-dihydropyrimidinone or amide group via a one-pot four-component reaction. Tetrahedron 69:881–887.
  26. 26. Alonso F, Moglie Y, Radivoy G, Yus M (2012) Multicomponent click synthesis of potentially biologically active triazoles catalyzed by copper nanoparticles on activated carbon in water. Heterocycles 84:1033–1044.
  27. 27. Shargi H, Beyzavi MS, Safavi A, Doroodman M, Mohammad M, et al. (2009) Immobilization of porphyniratocopper nanoparticles onto activated multi-walled carbon nanotubes and a study of its catalytic activity as an efficient heterogeneous catalyst for a click approach to the three-component synthesis of triazoles in water. Advnced Synthesis Catalysis 351:2391–2410.
  28. 28. Lee LV, Mitchell ML, Huang SJ, Fokin VV, Sharpless KB (2003) A potent and highly selective inhibitor of human α-1,3-Fucosyltransferase via click chemistry. Journal of American Chemical Society 125:9588–9589.
  29. 29. Gull Y, Rasool N, Noreen M, Nasim F-U-H, Yaqoob A, et al. (2013) Efficient synthesis of 2-amino-6-arylbenzothiazoles via Pd(0) Suzuki cross coupling reactions: potent urease enzyme inhibition and nitric oxide scavenging activities of the products. Molecules 18:8845–8857.
  30. 30. Serwar M, Akhtar T, Hameed S, Khan KM (2009) Synthesis, urease inhibition and antimicrobial activities of some chiral-5-aryl-4-(1-phenylpropyl)-2H-1,2,4-triazole-3(4H)-thiones. Arkivoc: 210–221.
  31. 31. Singh D, Mishra M, Gupta M, Singh P, Gupta A, et al. (2012) Original article nitric oxide radical scavenging assay of bioactive compounds present in methanol extract of Centella asiatica. International Journal of Pharmacy and Pharmaceutical Science Research 2:42–44.
  32. 32. Chippada SC, Vangalapati M (2011) Antioxidant, anti-inflammatory and anti arthritic activity of Centella asiatica extracts. Journal of Chemical, Biological and Physical Sciences 1:260–269.
  33. 33. Binder WH, Kluger C (2006) Azide/Alkyne-click reactions: Applications in material science and organic synthesis. Current Organic Chemistry 10:1791–1815.
  34. 34. Yadav JS, Reddy SBV, Reddy GM, Anjum RS (2009) Cu(OTf)2/Cu-catalyzed four-component reaction: A facile synthesis of α-alkoxytriazoles via click chemistry. Tetrahedron Letters 50:6029–6031.
  35. 35. Wu P, Feldman AK, Nugent AK, Hawker CJ, Scheel A, et al. (2004) Efficiency and fidelity in a click chemistry route to triazole dendrimers by the copper(I)-catalyzed ligation of azides and alkynes. Angewandte Chemie International Edition 43:3928–3932.
  36. 36. Mendoza-Espinosa D, Negron-Silva GE, Lomas-Romero L, Gutierrez-Carrillo A, Antillán R (2014) Pseudo-Four Component Synthesis of Mono- and Di-Benzylated-1,2,3-Triazoles Derived from Aniline. Molecules 19:55–66.
  37. 37. Sokmen BB, Onar HC, Yusufoglu A (2012) Anti-elastase, anti-urease and antioxidant activities of (3-13)-monohydroxyeicosanoic acid isomers. Journal of the Serbian Chemical Society 77:1353–1361.
  38. 38. Garrat DC (1964) The Quantitative analysis of Drugs. 3rd ed. Chapman & tall Japan.
  39. 39. Atta-ur-Rahman, Choudhary MI, Thomson WJ (2005) Bioassay techniques for drugs. Harward Academic Press. Amsterdam.