Next Article in Journal
Helping Eve Overcome ADAM: G-Quadruplexes in the ADAM-15 Promoter as New Molecular Targets for Breast Cancer Therapeutics
Previous Article in Journal
Effects of Aronia melanocarpa Constituents on Biofilm Formation of Escherichia coli and Bacillus cereus
Open AccessArticle

Synthesis and Biological Activity of 3-[Phenyl(1,3-thiazol-2-yl)-amino]propanoic Acids and Their Derivatives

1
Department of Organic Chemistry, Kaunas University of Technology, Radvilėnų pl. 19, LT-50254 Kaunas, Lithuania
2
Lithuanian University of Health Sciences, Veterinary Academy, Tilžės 18, LT-47181 Kaunas, Lithuania
3
Department of Food Technology, Kaunas University of Technology, Radvilėnų pl. 19, LT-50254 Kaunas, Lithuania
4
Lithuanian Research Centre for Agriculture and Forestry, Rumokai Experimental Station, Klausučiai, 70462 Vilkaviškis Distr., Lithuania
5
Institute of Agricultural and Food Sciences, Aleksandras Stulginskis University, Studentų g. 11, LT-53361 Akademija, Kaunor., Lithuania
*
Author to whom correspondence should be addressed.
Molecules 2013, 18(12), 15000-15018; https://doi.org/10.3390/molecules181215000
Received: 24 October 2013 / Revised: 3 December 2013 / Accepted: 3 December 2013 / Published: 5 December 2013
(This article belongs to the Section Organic Chemistry)

Abstract

New N,N-disubstituted β-amino acids and their derivatives with thiazole, aromatic, and heterocyclic substituents were synthesized from N-phenyl-N-thiocarbamoyl-β-alanine by the Hantzsch method; derivatives with hydrazone fragments were also obtained. Some of the synthesized compounds exhibited discrete antimicrobial activity, and 3-[(4-oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid was found to promote rapeseed growth and to increase seed yield and oil content.
Keywords: N-phenyl-N-thiocarbamoyl-β-alanine; Hantzsch synthesis; thiazole; hydrazone; biological activity N-phenyl-N-thiocarbamoyl-β-alanine; Hantzsch synthesis; thiazole; hydrazone; biological activity

1. Introduction

Recently, special attention has been paid to β-amino acids as potential starting materials for bio-organic, medicinal, and natural product chemistry [1]. Nitrogen-containing five- and six-membered heterocyclic compounds and their derivatives, which can be easily synthesized in laboratories are particularly important and often found in natural sources. Synthetic thiazole derivatives possess various biological activities such as anti-inflammatory [2,3], antipyretic [4,5], antiviral [6], anti-microbial [7,8,9,10,11], antifungal [12], and anticancer properties [13,14,15,16]. They have also been used as anticoagulants and antiarrhythmics [16], antidepressants [17], in the treatment of Alzheimer's disease, hypertension [18], schizophrenia [19], hepatitis C [20], allergies [21,22], HIV [23,24], diabetes [25,26], and in functional dye synthesis [27,28]. The 2-aminothiazole structure provides herbicidal [6,29] and antioxidant activities [30]. One of the most convenient methods of thiazole synthesis is the Hantzsch synthesis, which is based on condensation of α-haloketones or aldehydes with thioamides or thiourea. In our previous studies, N-aryl-N-thiocarbamoyl-β-alanines were used only for the synthesis of 1-substituted 2-thioxotetrahydro-4(1H)-pyrimidinones [31,32,33]. The aim of the present study was to synthesize new potentially biologically active N,N-disubstituted β-amino acids and their derivatives with thiazole, aromatic, heterocyclic fragments and to evaluate their chemical and biological properties.

2. Results and Discussion

2.1. Synthesis and Structural Peculiarities of New Compounds

3-[(4-Oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid (2) was obtained from N-phenyl-N-thiocarbamoyl-β-alanine (1) and monochloroacetic acid in different solvents (water, acetic acid, DMF, ethanol), and by using different bases such as sodium acetate, sodium carbonate, and triethylamine. Reactions in acetic acid and DMF were carried out at 90–100 °C and in ethanol and water, at the mixture boiling temperature for 5 h.
The studies showed that the best yield was obtained when reactions were carried out in water in the presence of sodium carbonate. Then the reaction mixture was acidified with acetic acid to pH 6. In order to obtain more thiazole derivatives and to assess the influence of substituents on the biological activity, a series of 3-{[5-substituted 4-oxo-4,5-dihydro-1,3-thiazol-2-yl](phenyl)amino}propanoic acids 3 with aromatic and heterocyclic fragments was synthesized (Scheme 1) . The syntheses were performed by condensation of dihydrothiazolone 2 and aromatic or heterocyclic aldehydes. Since the starting compound 2 and the resulting products 3 are soluble in an alkaline medium, the reactions were carried out in aqueous sodium carbonate solution in the presence of glycine as a bifunctional catalyst. The target products from the reaction mixtures were separated by acidification of the aqueous solutions with acetic acid to pH 6. The preliminary laboratory tests showed that the compounds 2 and 3 had growth regulating effects for rapeseed (Brassica napus L.) and St. John’s wort (Hypericum perforatum L.) [34]. In the same way, the compound 4 was synthesized from dihydrothiazolone 2 and terephthaldehyde.
Scheme 1. Synthesis of N-carboxyethylaminothiazoles and thiazolones 211.
Scheme 1. Synthesis of N-carboxyethylaminothiazoles and thiazolones 211.
Molecules 18 15000 g001
Reagents and Conditions: (i) ClCH2COOH, Na2CO3, H2O, reflux, 5 h, CH3COOH; (ii) RCHO, NH2CH2COOH, Na2CO3, H2O, reflux, 5 h, CH3COOH; (iii) terephthalaldehyde, NH2CH2COOH, Na2CO3, H2O, 5 h, CH3COOH; (iv) ClCH2CHO, H2O, reflux, 2 h, CH3COONa; (v) ClCH2COCH3, CO(CH3)2, reflux, 1 h, CH3COONa; (vi) 3-chloro-2,4-pentanedione, CO(CH3)2, CH3COONa, reflux, 2 h; (vii) 2-bromoacetophenone, CH3COONa, 2-propanol, reflux, 5 h; (viii) 2-bromo-2'-acetonaphthone, CH3COONa, 2-propanol, reflux, 4 h; (ix) 3-(bromoacetyl)coumarin, CH3COONa, 2-propanol, reflux, 7 h; (x) 2,3-dichloro-1,4-naphthoquinone, CH3COOH, CH3COONa, 80 °C, 24 h.
The structure of the synthesized compounds 3ai was confirmed by elemental analysis, 1H-NMR, 13C-NMR and IR spectral data. For example, in the 1H-NMR spectrum of compound 3a, the CH2CO and NCH2 group proton signals appear at 2.57 and 4.26 ppm, whereas aromatic and CH proton signals are observed in the range of 7.32 to 7.63 ppm. In the 13C-NMR spectrum of compound 3a, the signals of the CH2CO and NCH2 groups are observed at 34.57 and 51.44 ppm, respectively, whereas aromatic and CH= group carbon signals are observed in the range 111.96–140.42 ppm. The combination of CS, CN, CO, and COOH carbon signals are observed at 133.34, 171.94, 175.79, and 179.19 ppm respectively. In the IR spectrum, the C=N absorption band is observed at 1,540 cm1, two C=O bands at 1,686 and 1,725 cm1, and a band belonging to the OH group at 3,435 cm1 are observed. The formation of isomeric structures in these reactions was not observed. According to data published in the literature, glycine used as a bifunctional catalyst gives a single isomer (Z) [35].
3-[Phenyl(1,3-thiazol-2-yl)amino]propanoic acid (5) was synthesized from compound 1 and chloroacetaldehyde in refluxing water solution after 2 hours. A water-soluble amino acid hydrochloride is formed, which is present in the base transferred by the addition of sodium acetate to the reaction mixture. The structure of compound 5 is confirmed by its NMR spectral data: in the 1H-NMR spectrum two triplets at 2.62 and 4.11 ppm are assigned to the CH2CO and NCH2 group protons, while two doublets at 6.71 and 7.18 ppm belong to the thiazole ring and the SCH NCH group protons. Aromatic ring proton signals are observed at 7.31–7.50 ppm, and broad proton singlets of the carboxyl group are detectable at 12.30 ppm.
Reactions of 1 with various haloketones were also performed, and a number of new thiazole derivatives 611 with aliphatic, aromatic, or heterocyclic substituents were thus synthesized. The compounds 610 were synthesized in acetone or 2-propanol at boiling temperature. Cyclic diketones—quinones—are interesting both for their chemical properties and practical uses. Besides, many compounds containing the quinone group are found in Nature. Various assays showed that these compounds are characterized by a broad spectrum of biological activities [36]. A quinone fragment- containing compound 11 in this work was synthesized from N-phenyl-N-thiocarbamoyl-β-alanine (1) and 2,3-dichloro-1,4-naphthoquinone. The reaction was carried out in acetic acid at 80 °C for 24 h in the presence of sodium acetate. Then the mixture was cooled and diluted with water. The synthesized 3-(N-(4,9-dihydro-4,9-dioxonaphtho[2,3-d]thiazol-2-yl)-N-phenylamino)propanoic acid (11) was purified by dissolving it in aqueous sodium hydroxide solution, filtering, and then acidifying the filtrate with acetic acid to pH 6.
The compound 8 was modified by introducing a hydrazone fragment into its molecule with the porpose of increasing its antimicrobial properties (Scheme 2). Firstly, acid 8 was esterified with methanol to obtain the corresponding ester 12, which afterwards was boiled in 2-propanol with hydrazine monohydrate. The resulting acid hydrazide 13 was condensed with aromatic and heterocyclic aldehydes. The NMR spectra of the synthesized hydrazones 14 showed that in DMSO-d6 solution they exist as a mixture of E/Z isomers due the presence of a CO-NH fragment in the molecule and its hindered rotation around the CO-NH bond; however, the Z isomer was dominant.
Scheme 2. Synthesis of 3-[phenyl(4-phenyl-1,3-thiazol-2-yl)amino]propanoic acid derivatives 1214.
Scheme 2. Synthesis of 3-[phenyl(4-phenyl-1,3-thiazol-2-yl)amino]propanoic acid derivatives 1214.
Molecules 18 15000 g002
Reagents and Conditions: (i) CH3OH, H2SO4, reflux, 6 h; (ii) NH2NH2, 2-propanol, reflux, 7 h; (iii) RCHO, 2-propanol, reflux, 5 h.

2.2. Antimicrobial Evaluation of Synthesized Compounds

The antibacterial activity of the compounds was determined against Gram-positive spore-forming rods of Bacillus cereus (ATCC 11778), Gram-positive cocci of Staphylococcus (ATCC 9144), Gram-negative rods of E. coli (ATCC 8739) and Pseudomonas aeruginosa (NCTC 6750) by the broth and spread-plate methods. A range of concentrations for each compound was prepared according to the experimental procedure mentioned in experimental section. The minimal inhibition concentration (MIC, µg/mL) values are listed in Table 1.
Table 1. Antibacterial activity in vitro of the synthesized compounds (MIC, µg/mL).
Table 1. Antibacterial activity in vitro of the synthesized compounds (MIC, µg/mL).
CompoundStaphylococcus aureusBacillus cereusEsherichia coliPseudomonas aeruginosa
2nininini
3a250250250250
3b250250250250
3c250250250250
3d250250250250
3e250250250250
3f250250250250
3g250250250250
3hnininini
3i250250250250
5nininini
6250250250250
7250250250250
8250250250250
910005001000ni
10500500500500
11250250250250
Oxytetracycline62.562.562.562.5
ni—no inhibition. The minimum inhibitory concentrations were determined by the micro broth dilution method.
The MIC values of the samples were obtained in spread-plate tests, considering compound concentrations demonstrating antibacterial activity. Antibacterial screening data revealed a prominent inhibitory activity of compounds 3ag, and 3i against the tested strains of S. aureus, B. cereus,E. coli and P. aeruginosa (MIC = 250 µg/mL). The MIC value of 350 µg/mL was obtained against all bacteria for compounds 3a, 3b, 3c, 3d, 3f, 3e, 3i, 6, 7, 8, and 11, while for compounds 10 and 9, depending on bacterial species they were 500 or 1000 µg/mL. Compounds 2, 5 didn’t show antibacterial activity against the test microorganisms. Based on these data, antibacterial activity of the investigated compounds may be arranged in the following order:
Staphylococcus aureus:
Molecules 18 15000 i001
Bacillus cereus:
Molecules 18 15000 i002
Esherichia coli:
Molecules 18 15000 i003
Pseudomonas aeruginosa:
Molecules 18 15000 i004
The minimum bactericidal values (MBC) are presented in Table 2. These data also revealed pronounced inhibitory activity of some synthesized compounds against some bacteria species, particulalry S. aureus and B. cereus at 250 µg/mL. For instance, the lowest MBC values (250–500 µg/mL) were found for the compounds 3i, 3g, 11. However, the compounds 2, 3h, 5 didn’t show any bactericidal activity against the tested microorganisms.
Table 2. Bactericidal activity in vitro of the synthesized compounds (MBC, µg/mL)
Table 2. Bactericidal activity in vitro of the synthesized compounds (MBC, µg/mL)
CompoundStaphylococcus aureusBacillus cereusEsherichia coliPseudomonas aeruginosa
2nininini
3a5001000500500
3b500ni5001000
3c5001000500500
3d5001000500500
3e3501000500500
3f500ni500500
3g350350350350
3hnininini
3i250500500500
4ni50010001000
5nininini
6500ni500500
7500ni500500
8250ni500500
9100010001000ni
101000100010001000
11250250250250
14 bni100010001000
14 cni100010001000
14 ini100010001000
14 jni100010001000
Oxytetracycline62.562.5250250
ni—no inhibition; Minimum bactericidal concentrations were determined by using the spread plate method.
Based on these experiments, bactericidal activity of the investigated compounds may be arranged in the following order:
Decreasing MBC Values against Staphylococcus aureus:
Molecules 18 15000 i005
Decreasing MBC Values against Bacillus cereus:
Molecules 18 15000 i006
Decreasing MBC Values against Esherichia coli:
Molecules 18 15000 i007
Decreasing MBC Values against Pseudomonas aeruginosa:
Molecules 18 15000 i008
Among the all synthesized compounds, the highest antibacterial activity was exhibited by thiazole compound 11 containing a naphthoquinone ring. Among compounds 3ai, the highest antibacterial activity was shown by the compounds with furan and bromothiophene substituents in their structure. Aromatic substituents with different groups (NO2, F, Cl) didn’t show any inhibiting effect. Among thiazole compounds 59, the highest antibacterial activity was exhibited by compound 8 with a phenyl substituent in the 4-position of the thiazole ring.
Attempting to increase antimicrobial activity the structure of compound 8 was modified by replacing the carboxy group by hydrazone and hydrazide derivatives with aromatic and heterocyclic substituents, however, the results obtained suggest that modification of the carboxy group decreases the antimicrobial activity.
It may thus be concluded that the antimicrobial activity of the tested compounds depends on their structural peculiarities. 3-(N-(4,9-dihydro-4,9-dioxonaphtho[2,3-d]thiazol-2-yl)-N-phenylamino)propanoic acid (11), 3-(N-(5-((furan-2-yl)methylene)-4,5-dihydro-4-oxothiazol-2-yl)-N-phenylamino)propanoic acid (3g), and 3-(N-(5-((5-bromothiophen-2-yl)methylene)-4,5-dihydro-4-oxothiazol-2-yl)-N-phenyl- amino)propanoic acid (3i) were found to exhibit a promising antibacterial activity.

2.3. Evaluation of Synthesized Compounds on Rapeseed Yield and Biochemical Content

The effect of 3-[(4-oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid (2) on the rapeseed yield and biochemical content is presented in Table 3. When rapeseed seedlings were sprayed with solutions of 2 of various concentrations, the yield of seeds ranged from 2.07 to 2.44 t/ha, i.e., 19%–40% higher in comparison with the control sample. The highest seed yield was obtained when rapeseed seedlings were sprayed with 150 mg/L of the test compound solution. Other quality characteristics were also affected by spraying rapeseed seedlings with the test compound; for instance, the highest mass of 1,000 seeds (4.17 g) was determined when 25 mg/L were applied, whereas at the same concentration the oil content increased by 13.3%–39.3% in comparison with the control sample.
Table 3. The effect of 3-[(4-oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid (2) on rapeseed yield and biochemical content.
Table 3. The effect of 3-[(4-oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid (2) on rapeseed yield and biochemical content.
The concentration of 3-[(4-oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid (2), mg/L
0255075100125150
Yield, t/ha1.74 ± 0.112.07 ± 0.232.21 ± 0.082.11 ± 0.282.20 ± 0.082.41 ± 0.112.44 ± 0.20
Seed mass (per 1,000 seeds), g3.94 ± 0.164.17 ± 0.104.02 ± 0.123.81 ± 0.163.96 ± 0.093.85 ± 0.143.94 ± 0.21
Oil content, kg/t239.9 ± 1.7283.4 ± 8.3271.8 ± 7.5334.3 ± 5.7301.2 ± 4.9281.8 ± 7.9296.7 ± 6.5
Protein content, mg/100g15.9 ± 0.127.1 ± 0.330.7 ± 0.331.2 ± 0.330.3 ± 0.322.8 ± 0.422.5 ± 0.2
Ash, %4.53 ± 0.024.53 ± 0.054.48 ± 0.044.23 ± 0.104.16 ± 0.104.07 ± 0.144.10 ± 0.13
Application of 3-[(4-oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid (2) increased both the content of oil and protein. The highest content of oil (334 kg/t) and protein (31.2 mg/100g) was obtained upon using the 75 mg/L concentration of the study compound solution. The content of protein varied from 15.9 to 31.2 mg/100 g. When rapeseed seedlings were sprayed with 75 mg/L of the test compound solution, protein content was two times higher in comparison with the control sample.
The effect of 3-[(4-oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid (2) on the composition of fatty acids in rapeseed is presented in Table 4. The results showed that the variation of oleic acid content was insignificant (−0.09 to +0.77%) in comparison with the control sample. When rapeseed had been sprayed with 50 mg/L of the aforementioned solution, the highest content of oleic acid (61.51%) was obtained in the seeds. The content of saturated fatty acids (palmitic and stearic) varied insignificantly in comparison with the control sample. When rapeseed had been sprayed with the test compound solutions at various concentrations, some increase in the content of linolenic acid was observed; however, it was insignificant. Upon spraying rapeseed seedlings with 75 mg/L of the study compound solution, the highest content of linoleic acid (20.96%) was obtained in the seeds.
Table 4. The effect of 3-[(4-oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid (2) on the content of fatty acids in rapeseed.
Table 4. The effect of 3-[(4-oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid (2) on the content of fatty acids in rapeseed.
Fatty acid, %The concentration of 3-[(4-oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid (2), mg/L
0255075100125150
Palmitic5.11 ± 0.054.57 ± 0.064.79 ± 0.224.59 ± 0.075.23 ± 0.314.90 ± 0.094.76 ± 0.03
Stearic2.11 ± 0.082.02 ± 0.022.07 ± 0.012.02 ± 0.012.05 ± 0.012.04 ± 0.012.03 ± 0.01
Oleic61.04 ± 0.2661.07 ± 0.0961.51 ± 0.0361.28 ± 0.0260.98 ± 0.0861.21 ± 0.1861.41 ± 0.00
Linoleic20.55 ± 0.1320.83 ± 0.0920.61 ± 0.0120.96 ± 0.0620.93 ± 0.0220.84 ± 0.0520.85 ± 0.5
Eicosenoic1.02 ± 0.081.09 ± 0.031.07 ± 0.041.05 ± 0.010.93 ± 0.071.02 ± 0.020.99 ± 0.01
Linolenic8.49 ± 0.038.75 ± 0.028.54 ± 0.028.66 ± 0.028.67 ± 0.018.64 ± 0.018.60 ± 0.02
Summarizing the data of the field experiments spraying rapeseed seedlings with different concentrations of 3-[(4-oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid (2) solutions resulted in the following:
(a)
100 mg/L: the highest number of pods and siliques was obtained;
(b)
150 mg/L: the highest seed yield was obtained;
(c)
25 mg/L: the highest seed mass was obtained.
During osmotic stress plants accumulate in the cytoplasm of their cells organic osmolytes such as proline, valine, isoleucine, ectoine, aspartic acid, betaine, glucose, fructose, sucrose, fructans, mannitol, pinitol, and myo-inositol (inositol) [37]. The regulating effect of compound 2 on rapeseed could be explained by the properties of typical structure osmolytes: they could exist in the zwitterionic form (Scheme 3).
Scheme 3. The possible forms of compound 2.
Scheme 3. The possible forms of compound 2.
Molecules 18 15000 g003

3. Experimental

3.1. General

The starting materials and solvents were obtained from Sigma-Aldrich Chemie GmbH (Munich, Germany) and Fluka (Buchs, Switzerland) and were used without further purification. The methods used to follow the reactions were TLC and NMR. The NMR spectra were recorded on a Varian Unity Inova (300 MHz) spectrometer (Varian, Inc., Palo Alto, CA, USA) and Bruker Avance III/400 (400 MHz). Chemical shifts are expressed as δ, ppm relative to TMS. The J constants are given in Hz. The IR spectra (ν, cm−1) were recorded on a Perkin–Elmer BX FT–IR spectrometer (Perkin–Elmer Inc., Waltham, MA, USA) using KBr tablets. Mass spectra were obtained on a Waters ZQ 2000 spectrometer (Waters, Eschborn, Germany) using the electrospray ionization (ESI) mode and operating at 25 V. Elemental analyses were performed with a CE-440 elemental analyzer (Exeter Analytical Inc., Chelmsford, MA, USA). Melting points were determined with a B-540 Melting Point Analyzer (Büchi Corporation, New Castle, DE, USA) and are uncorrected. TLC was performed using Merck, Silica gel 60 F254 (Kieselgel 60 F254) plates.
3-[(4-Oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic Acid (2). A mixture of thioureido acid 1 (2.24 g, 0.01 mol), chloroacetic acid (1.04 g, 0.011 mol), sodium carbonate (1.16 g, 0.011 mol) and water (30 mL) was refluxed for 5 h. Then the mixture was acidified with acetic acid (6 mL) to pH 6. The precipitate was filtered off, washed with water, and crystallized. Yield 1.78 g, 68%; light yellow crystals m.p. 195–196 °C (from 2-propanol) (198–199 °C) [38]; IR (KBr): ν (cm−1) 3426 (OH), 1723, 1659 (CO); 1H-NMR (DMSO-d6): δ 2.58 (t, J = 7.4 Hz, 2H, CH2CO), 3.92 (s, 2H, SCH2), 4.17 (t, J = 7.3 Hz, 2H, NCH2), 7.41–7.57 (m, 5H, HAr), 12.42 (s, 1H, OH); 13C-NMR (DMSO-d6): δ 31.9 (CH2CO), 40.5 (SCH2), 49.9 (NCH2), 128.0, 129.6, 129.8, 140.1 (CAr), 171.9, 183.1 (CO), 186.9 (C=N); Anal. Calcd for C12H12N2O3S: C, 54.53%; H, 4.58%; N, 10.60%; Found: C, 54.69%; H, 4.56%; N, 10.68%.

3.2. General Procedure for Preparation of 3-{[(5Z)-5-Substituted 4-oxo-4,5-dihydro-1,3-thiazol-2-yl](phenyl)amino}propanoic Acids 3ai

A mixture of compound 2 (0.52 g, 2 mmol), the corresponding aldehyde (2.2 mmol), glycine (0.15 g, 2 mmol), and sodium carbonate (0.21 g, 2 mmol), water (6 mL) was refluxed for 5 h. Then the reaction mixture was diluted with water (50 mL) and acidified with acetic acid to pH 6. The precipitate was filtered and washed with water. Purification was performed by dissolving the crystals in 10% aqueous NaOH (100 mL), filtering, and acidifying the filtrate with acetic acid to pH 6 (the procedure was repeated 2 times).
3-{[(5Z)-5-Benzylidene-4-oxo-4,5-dihydro-1,3-thiazol-2-yl](phenyl)amino}propanoic Acid (3a). Yellow crystals, (0.52 g, 76%), m.p. 175–176 °C; IR (KBr): ν (cm−1) 3435 (OH), 1725, 1686 (CO), 1540 (CN); 1H-NMR (DMSO-d6): δ 2.57 (t, J = 7.4 Hz, 2H, CH2CO), 4.26 (t, J = 7.5 Hz, 2H, NCH2), 7.32–7.63 (m, 11H, HAr+CH); 13C-NMR (DMSO-d6): δ 32.1 (CH2CO), 50.1 (NCH2), 127.9, 128.9, 129.1, 129.5, 129.7, 130.1, 133.3, 139.6 (CAr, Cthiazole, CH), 172.0, 175.8 (CO), 179.2 (C=N); Anal. Calcd for C19H16N2O3S: C, 64.76%; H, 4.58%; N, 7.95%; Found: C, 64.44%; H, 4.63%; N, 8.04%.
3-{[(5Z)-5-(4-Nitrobenzylidene)-4-oxo-4,5-dihydro-1,3-thiazol-2-yl](phenyl)amino}propanoic Acid (3b). Yellow crystals, (0.57 g, 73%), m.p. 208–209 °C; IR (KBr): ν (cm−1) 3058 (OH), 1729, 1699 (CO), 1539 (CN); 1H-NMR (DMSO-d6): δ 2.34 (t, J = 7.8 Hz, 2H, CH2CO), 4.20 (t, J = 7.9 Hz, 2H, NCH2), 7.32–7.64 (m, 10 H, HAr+CH); 13C-NMR (DMSO-d6): δ 34.5 (CH2CO), 52.0 (NCH2), 115.7, 124.2, 127.8, 128.1, 130.3, 133.3, 139.6, 140.4, 147.0 (CAr, Cthiazole, CH), 172.2, 175.9 (CO), 179.0 (C=N); Anal. Calcd for C19H15N3O5S: C, 57.42%; H, 3.80%; N, 10.57%; Found: C, 56.94%; H, 3.75%; N, 10.86%.
3-{[(5Z)-5-(3-Nitrobenzylidene)-4-oxo-4,5-dihydro-1,3-thiazol-2-yl](phenyl)amino}propanoic Acid (3c). Yellow crystals, (0.50 g, 64%), m.p. 150–151 °C; IR (KBr): ν (cm−1) 3074 (OH), 1731, 1709 (CO), 1531 (CN); 1H-NMR (DMSO-d6): δ 2.59 (t, J = 7.5 Hz, 2H, CH2CO), 4.28 (t, J = 7.5 Hz, 2H, NCH2), 7.55–8.43 (m, 10H, HAr+CH); 13C-NMR (DMSO-d6): δ 32.7 (CH2CO), 50.9 (NCH2), 124.4, 128.5, 128.6, 130.6, 131.3, 132.4, 135.4, 135.9, 140.2, 148.6 (CAr, Cthiazole, CH), 172.6, 176.2 (CO), 179.6 (C=N); Anal. Calcd for C19H15N3O5S: C, 57.42%; H, 3.80%; N, 10.57%; Found: C, 57.39%; H, 3.77%; N, 10.51%.
3-{[(5Z)-5-(2-Chloro-4-nitrobenzylidene)-4-oxo-4,5-dihydro-1,3-thiazol-2-yl](phenyl)amino}propanoic Acid (3d). Yellow crystals, (0.53 g, 62%), m.p. 155–156 °C; IR (KBr): ν (cm−1) 3071 (OH), 1735, 1698 (CO), 1529 (CN); 1H-NMR (DMSO-d6): δ 2.63 (t, J = 7.4 Hz, 2H, CH2CO), 4.30 (t, J = 7.4 Hz, 2H, NCH2), 7.53–8.22 (m, 9H, HAr+CH); 13C-NMR (DMSO-d6): δ 31.9 (CH2CO), 50.4 (NCH2), 122.9, 123.6, 125.4, 128.2, 130.2, 130.3, 131.6, 133.4, 135.3, 139.6, 140.3, 146.6 (CAr, Cthiazole, CH), 172.0, 175.9 (CO), 178.6 (C=N); Anal. Calcd for C19H14ClN3O5S: C, 52.84%; H, 3.27%; N, 9.73%; Found: C, 52.78%; H, 3.33%; N, 9.78%.
3-{[(5Z)-5-(4-Fluorobenzylidene)-4-oxo-4,5-dihydro-1,3-thiazol-2-yl](phenyl)amino}propanoic Acid (3e). Yellow crystals, (0.62 g, 85%), m.p. 167–168 °C; IR (KBr): ν (cm−1) 3050 (OH), 1736, 1701 (CO), 1508 (CN); 1H-NMR (DMSO-d6): δ 2.61 (t, J = 7.2 Hz, 2H, CH2CO), 4.10 (t, J = 7.5 Hz, 2H, NCH2), 6.71–7.52 (m, 10H, HAr+CH), 12.29 (OH); 13C-NMR (DMSO-d6): δ 32.7 (CH2CO), 50.5 (NCH2), 116.1, 116.4, 128.1, 128.8, 129.1, 129.9, 130.0, 131.7, 139.9, 160.7 (CAr, Cthiazole, CH) 172.47, 175.8 (CO), 179.4 (C=N); Anal. Calcd for C19H15FN2O3S: C, 61.61%; H, 4.08%; N, 7.56%; Found: C, 61.77%; H, 3.91%; N, 7.48%.
3-{[(5Z)-4-oxo-5-(1H-Pyrrol-2-ylmethylene)-4,5-dihydro-1,3-thiazol-2-yl](phenyl)amino}propanoic Acid (3f). Yellow crystals, (0.46 g, 69%), m.p. 248–249 °C; IR (KBr): ν (cm−1) 3064 (OH), 1715, 1673 (CO), 1530 (CN); 1H-NMR (DMSO-d6): δ 2.28 (t, J = 8.3 Hz, 2H, CH2CO), 4.15 (t, J = 8.2 Hz, 2H, NCH2), 6.03–7.66 (m, 9H, HAr, CH, CHpyrrole), 11.79 (NH); 13C-NMR (DMSO-d6): δ 35.4 (CH2CO), 52.0 (NCH2), 111.2, 111.9, 120.5, 122.1, 123.3, 127.6, 128.3, 129.6, 130.0, 140.6 (CAr, Cthiazole, CH, Cpyrrole) 174.3, 174.7 (CO), 180.1 (C=N); Anal. Calcd for C17H15N3O3S: C, 59.81%; H, 4.43%; N, 12.31%; Found: C, 59.75%; H, 4.39%; N, 12.22%.
3-{[(5Z)-5-(2-Furylmethylene)-4-oxo-4,5-dihydro-1,3-thiazol-2-yl](phenyl)amino}propanoic Acid (3g). Brown crystals, (0.41 g, 61%), m.p. 139–140 °C; IR (KBr): ν (cm−1) 3050 (OH), 1727, 1682 (CO), 1532 (CN); 1H-NMR (DMSO-d6): δ 2.61 (t, J = 7.4 Hz, 2H, CH2CO), 4.25 (t, J = 7.4 Hz, 2H, NCH2), 6.61–7.91 (m, 9H, HAr, CH, Hfuran); 13C-NMR (DMSO-d6): δ 32.0 (CH2CO), 49.8 (NCH2), 113.3, 117.0, 117.2, 126.4, 128.3, 130.0, 130.1, 140.0, 146.6, 149.5 (CAr, Cthiazole, CH, Cfuran), 172.1, 176.8 (CO), 179.4 (C=N); Anal. Calcd for C17H14N2O4S: C, 59.64%; H, 4.12%; N, 8.18%; Found: C, 59.58% H, 4.21%; N, 8.22%.
3-{[(5Z)-4-Oxo-5-(2-thienylmethylene)-4,5-dihydro-1,3-thiazol-2-yl](phenyl)amino}propanoic Acid (3h). Yellow crystals (0.48 g, 68%), m.p. 137–138 °C; IR (KBr): ν (cm−1) 3057 (OH), 1726, 1678 (CO), 1542 (CN); 1H-NMR (DMSO-d6): δ 2.62 (t, J = 7.4 Hz, 2H, CH2CO), 4.26 (t, J = 7.4 Hz, 2H, NCH2), 7.16–7.89 (m, 9H, HAr+CH, Hthiophene); 13C-NMR (DMSO-d6): δ 32.0 (CH2CO), 50.0 (NCH2), 123.5, 127.2, 128.3, 128.9, 130.0, 130.1, 131.5, 133.5, 138.4, 140.0 (CAr, Cthiazole, CH, Cthiophene), 172.1, 175.3 (CO), 179.3 (C=N); Anal. Calcd for C17H14N2O3S2: C, 56.96%; H, 3.94%; N, 7.82%; Found: C, 57.02%; H, 3.88%; N, 7.89%.
3-[{(5Z)-5-[(5-Bromo-2-thienyl)methylene]-4-oxo-4,5-dihydro-1,3-thiazol-2-yl}(phenyl)amino]propanoic Acid (3i). Yellow crystals (0.47 g, 55%), m.p. 143–144 °C; IR (KBr): ν (cm−1) 3007 (OH), 1723, 1682 (CO), 1538 (CN); 1H-NMR (DMSO-d6): δ 2.57 (t, J = 7.5 Hz, 2H, CH2CO), 4.25 (t, J = 7.5 Hz, 2H, NCH2), 7.28–7.84 (m, 8H, HAr, CH, Hthiophene); 13C-NMR (DMSO-d6): δ 32.4 (CH2CO), 50.4 (NCH2), 116.9, 122.41, 124.4, 128.0, 128.2, 130.1, 132.3, 133.8, 139.9, 140.2 (CAr, Cthiazole, CH, Cthiophene), 172.4, 174.8 (CO), 179.0 (C=N); Anal. Calcd for C17H13BrN2O3S2: C, 46.69%; H, 3.00%; N, 6.41%; Found: C, 46.77%; H, 2.97%; N, 6.50%.
3-({5-(4-{[2-[(3-Hydroxy-3-oxopropyl)aniline]-4-oxo-1,3-thiazol-5-yliden]methyl}phenyl)methyliden]-4-oxo-4,5-dihydro-1,3-thiazol-2-yl}aniline)propanoic Acid (4). A mixture of compound 2 (0.61 g, 2.3 mmol), terephthalaldehyde (0.15 g, 1.14 mmol), glycine (0.17 g, 2.3 mmol), sodium carbonate (0.49 g, 4.6 mmol) and water (7 mL) was refluxed for 5 h. Then the reaction mixture was diluted with water (50 mL) and acidified with acetic acid to pH 6. The precipitate was filtered and washed with water. Purification was performed by dissolving the crystals in 2% aqueous KOH (100 mL), filtering, and acidifying the filtrate with acetic acid to pH 6. Yellow crystals, (0.34 g, 27%), m.p. 269–270 °C; IR (KBr): ν (cm−1) 3023 (OH), 1729, 1710, 1694, 1688 (CO), 1532, 1538 (CN); 1H-NMR (DMSO-d6): δ 2.61 (t, J = 7.2 Hz, 4H, CH2CO), 4.27 (t, J = 7.1 Hz, 4 H, NCH2), 7.46–7.65 (m, 16H, CAr, CH); Anal. Calcd for C32H26N4O6S2: C, 61.33%; H, 4.18%; N, 8.94%; Found: C, 61.30%; H, 4.07%; N, 8.88%.
3-[Phenyl(1,3-thiazol-2-yl)amino]propanoic Acid (5). A mixture of the thioureido acid 2 (1.12 g, 5 mmol), chloroacetaldehyde 50% aqueous solution (0.79 g, 0.01 mol), water (20 mL) was refluxed for 2 h. Then sodium acetate (0.82 g, 0.01 mol) was added and the mixture was stirred for 5 min. The precipitate was filtered and washed with water. Purification was performed by dissolving the crystals in 10% aqueous sodium carbonate (75 mL), filtering, and acidifying the filtrate with acetic acid to pH 6 (the procedure was repeated 2 times). Yellow crystals, (0.50 g, 42%), m.p. 144–145 °C; IR (KBr): ν (cm−1) 3076 (OH), 1717 (CO), 1514 (CN); 1H-NMR (DMSO-d6): δ 2.62 (t, J = 7.2 Hz, 2H, CH2CO), 4.11 (t, J = 7.5 Hz, 2H, NCH2), 6.71 (d, J = 3.6 Hz, 1H, NCH), 7.18 (d, J = 3.6 Hz, 1H, SCH), 7.31–7.50 (m, 5H, HAr), 12.30 (br s, 1H, OH); 13C-NMR (DMSO-d6): δ 32.2 (CH2CO), 48.5 (NCH2), 108.1, 126.6, 127.2, 130.0, 139.1, 144.8 (CAr, Cthiazole) 169.5 (CO), 172.5 (C=N); Anal. Calcd for C12H12N2O2S: C, 58.05%; H, 4.87%; N, 11.28%; Found: C, 58.11%; H, 4.93%; N, 11.31%.
3-[(4-Methyl-1,3-thiazol-2-yl)(phenyl)amino]propanoic Acid (6). A mixture of thioureido acid 1 (0.56 g, 2.5 mmol), chloroacetone (0.29 g, 3 mmol) in acetone (10 mL) was refluxed for 1 h and diluted with water (50 mL). Then sodium acetate (0.98 g, 12 mmol) was added, and the mixture was stirred for 5 min. The precipitate was filtered, and washed with water. Yellow crystals (0.52 g, 79%), m.p. 149–150 °C (from acetone-water mixture); IR (KBr): ν (cm−1) 3128 (OH), 1712 (CO), 1515 (CN); 1H-NMR (DMSO-d6): δ 2.16 (s, 3H, CH3), 2.60 (t, J = 7.3 Hz, 2H, CH2CO), 4.09 (t, J = 7.4 Hz, 2H, NCH2), 6.26 (s, 1H, SCH), 7.30–7.49 (m, 5H, HAr), 12.29 (br s, 1H, OH); 13C-NMR (DMSO-d6): δ 17.4 (CH3), 32.3 (CH2CO), 48.3 (NCH2), 102.1, 126.8, 127.2, 129.9, 144.5, 148.3 (CAr, Cthiazole); 168.67 (CO), 172.6 (C=N); Anal. Calcd for C13H14N2O2S: C, 59.52%; H, 5.38%; N, 10.68%; Found: C, 59.47%; H, 5.45%; N, 10.55%.
3-[(5-Acetyl-4-methyl-1,3-thiazol-2-yl)(phenyl)amino]propanoic Acid (7). A mixture of thioureido acid 1 (1.0 g, 4.5 mmol), 3-chloro-2,4-pentanedione (0.76 g, 5.6 mmol), acetone (10 mL) was refluxed for 2 h and diluted with water (50 mL). Then sodium acetate ((1.8 g, 22 mmol) was added, and the mixture was stirred for 5 min. The precipitate was filtered and washed with water. Purification was performed by dissolving the crystals in 10% aqueous sodium hydroxide (50 mL), filtering, and acidifying the filtrate with acetic acid to pH 6. Light yellow crystals, (0.84 g, 92%), m.p. 175–176 °C; IR (KBr): ν (cm−1) 1721, 1700 (CO), 1576 (C=N); 1H-NMR (DMSO-d6): δ 2.31 (s, 3H, CH3), 2.48 (s, 3H, CH3), 2.51 (t, J = 7.2 Hz, 2H, CH2CO), 4.15 (t, J = 7.3 Hz, 2H, NCH2), 7.44–7.53 (m, 6H, HAr, Hthiazole), 12.36 (s, 1H, OH); Anal. Calcd for C15H16N2O3S: C, 59.19%; H, 5.30%; N, 9.20%; Found: C, 59.26%; H, 5.15%; N, 9.11%.
3-[Phenyl(4-phenyl-1,3-thiazol-2-yl)amino]propanoic Acid (8). A mixture of thioureido acid 2 (1.12 g, 5 mmol), 2-bromoacetophenone (0.99 g, 5 mmol), sodium acetate (0.82 g, 10 mmol), 2-propanol (10 mL) was refluxed for 5 h. The mixture was diluted with water (30 mL). The precipitate was filtered and washed with water. Purification was performed by dissolving the crystals in 10% aqueous KOH (30 mL), filtering, and acidifying the filtrate with acetic acid to pH 6. Yellow crystals, (1.56 g, 96%), m.p. 101–102 °C; IR (KBr): ν (cm−1) 3416 (OH), 1712 (CO), 1508 (CN); 1H-NMR (DMSO-d6): δ 2.66 (t, J = 7.3 Hz, 2H, CH2CO), 4.19 (t, J = 7.5 Hz, 2H, NCH2), 7.13 (s, 1H, SCH), 7.26–7.88 (m, 10H, HAr); 13C-NMR (DMSO-d6): δ 32.8 (CH2CO), 48.9 (NCH2), 102.9, 125.6, 126.7, 127.3, 127.5, 128.5, 129.9, 134.6, 150.3, 144.6 (CAr, Cthiazole), 168.7 (CO), 173.0 (C=N); Anal. Calcd for C18H16N2O2S: C, 66.64%; H, 4.97%; N, 8.64%; Found: C, 66.60%; H, 4.83%; N, 8.57%.
3-{[4-(2-Naphthyl)-1,3-thiazol-2-yl](phenyl)amino}propanoic Acid (9). A mixture of thioureido acid 2 (1.12 g, 5 mmol), 2-bromoacetonaphthone (1.25 g, 5 mmol), 2-propanol (10 mL), sodium acetate (0.82 g, 10 mmol) was refluxed for 4 h. The mixture was diluted with water. The precipitate was filtered and washed with water. Purification was performed by dissolving the crystals in 10% aqueous K2CO3 (50 mL), filtering, and acidifying the filtrate with acetic acid to pH 6. Yellow crystals, (1.52 g, 82%), m.p. 143–144 °C; IR (KBr): ν (cm−1) 3112 (OH), 1741 (CO), 1511 (CN); 1H-NMR (DMSO-d6): δ 2.67 (t, J = 7.4 Hz, 2H, CH2CO), 4.25 (t, J = 7.4 Hz, 2H, NCH2), 7.30 (s, 1H, SCH), 7.48–7.88 (m, 12H, HAr); Anal. Calcd for C22H18N2O2S: C, 70.57%; H, 4.85%; N, 7.48%; Found: C, 70.41%; H, 4.92%; N, 7.39%.
3-{[4-(2-Oxo-2H-chromen-3-yl)-1,3-thiazol-2-yl](phenyl)amino}propanoic Acid (10). A mixture of thioureido acid 2 (1.12 g, 5 mmol), 3-(bromoacetyl)coumarin (1.34 g, 5 mmol), sodium acetate (0.82 g, 10 mmol), 2-propanol (10 mL) was refluxed for 7 h. After cooling, the precipitate was filtered and washed with water. Purification was performed by dissolving the crystals in 10% aqueous KOH (50 mL), filtering, and acidifying the filtrate with acetic acid to pH 6. Light brown crystals (1.68 g, 86%), m.p. 172–173 °C; IR (KBr): ν (cm−1) 3137 (OH), 1717, 1709 (CO), 1536 (CN); 1H-NMR (DMSO-d6): δ 2.65 (t, J = 7.3 Hz, 2H, CH2CO), 4.23 (t, J = 7.3 Hz, 2H, NCH2), 7.34–7.89 (m, 10H, HAr), 8.64 (s, 1H, SCH); Anal. Calcd for C21H18N2O4S: C, 63.95%; H, 4.60%; N, 7.10%; Found: C, 64.11%; H, 4.66%; N, 7.08%.
3-(N-(4,9-Dihydro-4,9-dioxonaphtho[2,3-d]thiazol-2-yl)-N-phenylamino)propanoic Acid (11). A mixture of thioureido acid 2 (2.24 g, 0.01 mol), 2,3-dichloro-1,4-naphthoquinone (2.72 g, 12 mmol), sodium acetate (2.95 g, 36 mmol), acetic acid (50 mL) was stirred at 80 °C for 24 h. After cooling mixture was diluted with water (100 mL). The precipitate formed was filtered and washed with water. Purification was performed by dissolving the crystals in 10% aqueous KOH (350 mL), filtering, and acidifying the filtrate with acetic acid to pH 6. Claret crystals (2.38 g, 62%), m.p. 185 °C (decomp.); IR (KBr): ν (cm−1) 3063 (OH), 1712, 1634, 1622 (CO), 1527 (CN); 1H-NMR (DMSO-d6): δ 2.71 (t, J = 7.1 Hz, 2H, CH2CO), 4.53 (t, J = 7.1 Hz, 2H, NCH2), 7.43–7.91 (m, 9H, HAr), 12.19 (br s, 1H, OH); 13C-NMR (DMSO-d6): δ 32.1 (CH2CO), 49.1 (NCH2), 120.6, 125.8, 127.0, 128.7, 129.3, 130.5, 131.7, 135.0, 142.6, 160.1 169.8 (CAr, Cthiazole), 172.1, 175.3, 179.8 (CO); Anal. Calcd for C20H14N2O4S: C, 63.48%; H, 3.73%; N, 7.40%; Found: C, 63.36%; H, 3.62%; N, 7.44%.
Methyl 3-[phenyl(4-phenyl-1,3-thiazol-2-yl)amino]propanoate (12). A mixture of 3-(N-phenyl-N-(4-phenylthiazol-2-yl)amino)propanoic acid (8) (8.08 g, 0.025 mol), methanol (150 mL), and H2SO4 (2 mL) was refluxed for 6 h. Then the solvent was evaporated. The precipitated product was neutralized with 5% sodium bicarbonate solution and extracted with diethyl ether (3 times with 100 mL); then the ether was evaporated. Oil, (7.5 g, 89%), Rf = 0.7; IR (KBr): ν (cm−1) 1737 (CO), 1532 (CN); 1H-NMR (DMSO-d6): δ 2.89 (t, J = 7.2 Hz, 2H, CH2CO), 3.61 (s, 3H, CH3), 4.35 (t, J = 7.2 Hz, 2H, NCH2), 6.67 (s, 1H, SCH), 7.24–7.90 (m, 10H, HAr); 13C-NMR (DMSO-d6): δ 32.9 (CH2CO), 49.1 (NCH2), 51.8 (CH3), 101.9, 126.1, 127.1, 127.6, 127.7, 128.6, 130.2, 135.1, 145.1, 151.5 (CAr, Cthiazole), 169.4 (CO), 172.4 (C=N); Anal. Calcd for C19H18N2O2S: C, 67.43%; H, 5.36%; N, 8.28%; Found: C, 67.35%; H, 5.42%; N, 8.19%.
3-[Phenyl(4-phenyl-1,3-thiazol-2-yl)amino]propanehydrazide (13). A mixture of ester 12 (7.2 g, 0.02 mol), hydrazine monohydrate (2.07 g, 0.04 mol) was refluxed in 2-propanol (20 mL) for 7 h. The reaction mixture was cooled, the solvent was evaporated, and the oilseed substance was purified by chromatography on a silica gel 60 column (methanol–chloroform, 1:20), Rf 0.6. Oil, (5.1 g, 71%), IR (KBr): ν (cm−1) 3283, 3060 (NH, NH2), 1661 (CO), 1532 (CN); 1H-NMR (DMSO-d6): δ 2.71 (t, J = 6.7 Hz, 2H, CH2CO), 3.53–4.06 (br s, 2H, NH2), 4.34 (t, J = 6.7 Hz, 2H, NCH2), 6.67 (s, 1H, SCH), 7.26–7.88 (m, 10H, HAr), 8.06 (s, 1H, NH); 13C-NMR (DMSO-d6): δ 33.6 (CH2CO), 49.4 (NCH2), 102.3, 126.1, 126.9, 127.8, 127.9, 128.8, 130.3, 134.8, 144.7, 151.3 (CAr, Cthiazole), 170.1 (CO), 171.7 (C=N); Anal. Calcd for C18H18N4OS: C, 63.88%; H, 5.36%; N, 16.56%; Found: C, 63.72%; H, 5.45%; N, 16.59%.

3.3. General Synthesis of Hydrazones 14 b, c, i, j

A mixture of hydrazide 13 (1.01 g, 3 mmol), the corresponding aldehyde (4 mmol) and 2-propanol (6 mL) was refluxed for 5 h. After cooling, the precipitated compound was filtered, washed with 2-propanol, and crystallized.
3-[(4-Phenyl-1,3-thiazol-2-yl)(phenyl)amino]-N'-(4-nitrophenyl)methylene]propanehydrazide (14b). Yellow crystals, (1.19 g, 84%), m.p. 218–219 °C (from 1,4-dioxane); IR (KBr): ν (cm−1) 3082 (NH), 1676 (CO), 1541, 1514 (CN); 1H-NMR (DMSO-d6): δ (E/Z, 40/60): 2.76, 3.18 (2 t, J = 7.1 Hz, 2H, CH2CO), 4.27–4.38 (m, 2H, NCH2), 7.13–8.30 (m, 15H, HAr, Htiazole), 8.25, 8.30 (2 s, 1H, N=CH), 11.70, 11.78 (1 H, 2 s, NHN); Anal. Calcd for C25H21N5O3S: C, 63.68%; H, 4.49%; N, 14.85%; Found: C, 63.55%; H, 14.77%; N, 8.50%.
3-[(4-Phenyl-1,3-thiazol-2-yl)(phenyl)amino]-N'-(3-nitrophenyl)methylene]propanehydrazide (14c). Red-yellow crystals, (1.15 g, 81%), m.p. 147–148 °C (from 1,4-dioxane); IR (KBr): ν (cm−1) 3109 (NH), 1681 (CO), 1532, 1515 (CN); 1H-NMR (DMSO-d6): δ (E/Z, 40/60): 2.75, 3.15 (2 t, J = 7.1 Hz, 2H, CH2CO), 4.27–4.39 (m, 2H, NCH2), 7.10–8.36 (m, 15H, HAr, Hthiazole), 8.47, 8.50 (2 s, 1H, N=CH), 11.62, 11.73 (2 s, 1H, NHN); Anal. Calcd for C25H21N5O3S: C, 63.68%; H, 4.49%; N, 14.85%; Found: C, 66.79%; H, 4.61%; N, 14.80%.
N'-[(5-Bromo-2-thienyl)methylene]-3-[(4-phenyl-1,3-thiazol-2-yl)(phenyl)amino]propanehydrazide (14i). Grey crystals, (1.1 g, 72%), m.p. 144–145 °C (from 1,4-dioxane); IR (KBr): ν (cm−1) 3057 (NH), 1674 (CO), 1533, 1514 (CN); 1H-NMR (DMSO-d6): δ (E/Z, 40/60): 2.69, 3.04 (2 t, J = 7.1 Hz, 2H, CH2CO), 4.23–4.34 (m, 2H, NCH2), 7.09–7.93 (m, 13H, HAr, Htiazole, Hthiophene), 8.04, 8.28 (2 s, 1H, N=CH), 11.43, 11.49 (2 s, 1H, NNH); Anal. Calcd for C23H19BrN4OS2: C, 54.01%; H, 3.74%; N, 10.95%; Found: C, 54.15%; H, 3.82%; N, 11.03%.
N'-[(5-Nitro-2-thienyl)methylene]-3-[(4-phenyl-1,3-thiazol-2-yl)(phenyl)amino]propanehydrazide (14j). Orange crystals, (0.98 g, 70%), m.p. 208–209 °C (from 1,4-dioxane); IR (KBr): ν (cm−1) 3065 (NH), 1672 (CO), 1541, 1518 (CN); 1H-NMR (DMSO-d6): δ (E/Z, 40/60): 2.74, 3.08 (2 t, J = 7.0 Hz, 2H, CH2CO), 4.25–4.37 (m, 2H, NCH2), 7.11–8.11 (m, 13H, HAr, Hthiazole, Hthiophene), 8.12, 8.38 (2 s, 1H, N=CH), 11.79, 11.83 (2 s, 1H, NNH); Anal. Calcd for C23H19N5O3S2: C, 57.85%; H, 4.01%; N, 14.66%; Found: C, 57.77%; H, 4.14%; N, 14.71%.

3.4. Antimicrobial Activity

The following bacteria strains were used: gram-negative rods of Escherichia coli (ATCC 8739), Pseudomonas aeruginosa (NCTC 6750), gram-positive cocci of Staphylococcus aureus (ATCC 9144) and gram-positive spore forming rods of Bacillus cereus (ATCC 11778). Tryptic soy agar (TSA) and tryptic soy broth (TSB) were used for bacteria cultivation and antibacterial activity tests.
Antibacterial activity of the compounds was determined by testing their different concentrations against B. cereus, S. aureus, P. aeruginosa and E. coli bacteria by the broth-dilution and spread plate methods [39,40]. A range of concentrations, 1000, 500, 350, 250, and 125 µg/mL were prepared for each sample. They were streaked out on TSA plates and incubated at 37 °C for 24 h. A representative colony was placed in 5 mL of TBS and incubated at 37 °C for 24 h.
B. cereus, S. aureus, E. coli, and P. aeruginosa cultures containing 108 CFU/mL (colony-forming units corresponding to McFarland’s 0.5) were diluted with TSB and used for the antibacterial test. The test organisms (100 µL) were added to each tube and incubated at 37 °C for 24 h. At the end of this period, a small amount of the diluted mixture (different materials) from each tube was pulled out and spread on TSA. The plates were incubated at 37 °C for 48 h. The growth of bacterial cells was observed on agar plates. The lowest concentration of the bacterial material at which no growth was observed was considered as the minimum bactericidal concentration (MBC) value [41]. The antibacterial compound oxytetracycline inoculated with each test bacterium in the tubes and plates was used as a control. The growth of the test bacteria was observed in all plates as positive controls.

3.5. Field Trials and Rapeseed Biochemical Ontent

The influence of N-substituted β-alanine derivatives on the growth of rapeseed (Brassica napus L.) and St. John wort (Hypericum perforatum L.) in vitro was explored. According to the results, 3-[(4-oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid (2) was selected, because it had the most remarkable positive effect on the growth of rapeseed and St. John wort in vitro [34]. Following the data obtained during the in vitro experiments, compound 2 was selected for the field trials.
Experiments with 3-[(4-oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid on spring rapeseed (Brassica napus L.) of the SW Landmark variety were carried out in field trials in 2012 at the Lithuanian Research Centre for Agriculture and Forestry branch Rumokai experimental station. Spring rapeseed was sprayed with the study compound solutions (concentrations 25–150 mg/L) before flowering. For each field, 1 L of the solution was used. The soil of the experimental field was carbonaceous shallow luvisols (Calc or epihypogleyic luvisol IDg8-k (LVg-w-cc) with the following charateristics: pHKCl 6.7, humus 1.82%, Ntotal 0.12%, 308 mg kg−1 of mobile P2O5, 296 mg kg−1 of mobile K2O. Soil samples were collected on 25.04.2012. Barley was the preceding crop. The size of the initial and accounted fields was 2.7 × 11 m and 2.2 × 10 m, respectively. Experiments with rapeseed were performed in April–August. The average air temperature ranged from 7.9 to 19.3 °C, the rainfall ranged from 44.4 to 118.4 mm per month, and during the experiment it was 361.1 mm. The rainfall was by 11.1 mm higher than the average multi-annual precipitation. Biometric measurements were carried out after harvesting. The moisture of rape seeds was 8.5%. The oil content in rape seeds was measured by extraction with hexane for 3 hours in a Soxhlet apparatus (Behr Labor-Technik, Dusseldorf, Germany). Fatty acid composition was analysed with a HRGC 5300 Mega Series gas chromatograph Carlo Erba Strumentazione (Milano, Italy). The protein content in rape seeds was measured by the Bradford method.

4. Conclusions

The N-phenyl-N-thiocarbamoyl-β-alanine was used for the first time for the synthesis of biologically active 2-aminothiazole derivatives. By a new method of synthesizing the target structures we have established a convenient way to produce a number of compounds, which were used for biological testing. Some of the synthesized compounds exhibited promising antimicrobial activity, and 3-[(4-oxo-4,5-dihydro-1,3-thiazol-2-yl)(phenyl)amino]propanoic acid (2) was found to promote the growth, to increase seed yield and oil content in rapeseed.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Juaristi, E.; Soloshonok, V.A. Enantioselective Synthesis of β-amino Acids (E), 2nd ed.; John Wiley & Sons, Inc.: Haboken, NJ, USA, 2005; pp. 477–614. [Google Scholar]
  2. Kouatly, O.; Geronikaki, A.; Kamoutsis, C.; Hadjipavlou-Litina, D.; Eleftheriou, P. Adamantane derivatives of thiazolyl-N-substituted amide, as possible non-steroidal anti-inflammatory agents. Eur. J. Med. Chem. 2008, 20, 1–7. [Google Scholar]
  3. Carradori, S.; Secci, D.; Bolasco, A.; De Monte, C.; Yáñez, M. Synthesis and selective inhibitory activity against human COX-1 of novel 1-(4-substituted-thiazol-2-yl)-3,5-di(hetero)aryl-pyrazoline derivatives. Arch. Pharm. 2012, 345, 973–979. [Google Scholar] [CrossRef]
  4. Holla, B.S.; Malini, K.V.; Rao, B.S.; Sarojini, B.K.; Kumari, N.S. Synthesis of some new 2,4-disubstituted thiazoles as possible antibacterial and anti-inflammatory agents. Eur. J. Med. Chem. 2003, 38, 313–318. [Google Scholar] [CrossRef]
  5. Toyoshi, K.; Misako, A.; Masako, K.; Katsuya, O.; Shigekatsu, K.; Tamatsu, M.; Takeshi, I. Syntheses and antiinflammatory activity of malonamic acid, malonamate and malonamide derivatives of some heterocyclic compounds. Chem. Pharm. Bull. 1985, 33, 4878–4888. [Google Scholar] [CrossRef]
  6. Dovlatyan, V.V.; Eliazyan, K.A.; Pivazyan, V.A.; Kazaryan, E.A.; Engoyan, A.P. Thiazolecarboxylic acid derivatives. 1. n-substituted 2-amino-4-methylthiazole-5-carboxylic acid derivatives. Chem. Heterocycl. Compd. 2004, 40, 84–89. [Google Scholar] [CrossRef]
  7. Zitouni, G.; Demirayak, S.; Ozdemir, A.; Kaplancikli, Z.; Yildiz, M. Synthesis of some 2-[(benzazole-2-yl)thioacetylamino]thiazole derivatives and their antimicrobial activity and toxicity. Eur. J. Med. Chem. 2004, 39, 267–272. [Google Scholar] [CrossRef]
  8. Argyropoulou, I.; Gerinikaki, A.; Vicini, P.; Zani, F. Synthesis and biological evaluation of sulfonamide thiazole and benzothiazole derivatives as antimicrobial agents. Arkivoc 2009, 18, 89–102. [Google Scholar] [CrossRef]
  9. Prakasha, K.C.; Raghavendra, G.M.; Harisha, R.; Gowda, D.C. Design, Synthesis and antimicrobial screening of amino acids conjugated 2-amino-4-arylthiazole derivatives. Int. J. Pharm. Sci. 2011, 3, 120–125. [Google Scholar]
  10. Price, K.E.; Chisholm, D.R.; Godfrey, J.C.; Misiek, M.; Gourevitch, A. Semisynthetic coumermycins: Structure-activity relationships. Appl. Microbiol. 1970, 19, 14–26. [Google Scholar]
  11. Nair, S.; Garg, S.P.; Sah, P. Synthesis and antibicrobial activity of some new oxazolone derivatives of 4,5-disubstituted-2-aminothiazoles. J. Indian Chem. Soc. 2006, 83, 205–207. [Google Scholar]
  12. Qin, X.; Yu, H.; Liu, J.; Dai, H.; Bing, G.; Qin, Z.; Zhang, X.; Wang, T.; Fang, J. Synthesis and biological activity of novel N-(5-((1H-1,2,4-triazol-1-yl)methyl)-4-tert-butylthiazol-2-yl)-4-carboxamide derivatives. Arkivoc 2009, 2, 201–210. [Google Scholar]
  13. Chen, B.C.; Zhao, R.; Wang, B.; Droghini, R.; Lajeunesse, J.; Sirard, P.; Endo, M.; Balasubramanian, B.; Barrish, J.C. A new and efficient preparation of 2-aminothiazole-5-carbamides: applications to the synthesis of the anti-cancer drug dasatinib. Arkivoc 2010, 6, 32–38. [Google Scholar]
  14. Al-Saadi, M.S.; Faidallah, H.M.; Rostom, S.A. Synthesis and biological evalution of some 2,4,5-trisubstituted thiazole derivatives as potencial antimicrobial and anticancer agents. Arch. Pharm. Chem. Life Sci. 2008, 341, 624–634. [Google Scholar] [CrossRef]
  15. Wityak, J.; Das, J.; Moquin, R.V.; Shen, Z.; Lin, J.; Chen, P.; Doweyko, A.M.; Pitt, S.; Pang, S.; Shen, D.R.; et al. Discovery and initial SAR of 2-amino-5-carboxamidothiazoles as inhibitors of the Src-family kinase p56Lck. Bioorg. Med. Chem. Lett. 2003, 3, 4007–4010. [Google Scholar]
  16. Amr, A.E.; Sabrry, N.M.; Abdalla, M.M.; Abdel-Wahab, B.F. Synthesis, Antiarrhytmic and anticoagulant activities of novel thiazolo derivatives from methyl 2-(thiazol-2-ylcarbamoyl)acetate. Eur. J. Med. Chem. 2009, 44, 725–735. [Google Scholar] [CrossRef]
  17. Harnett, J.J.; Roubert, V.; Dolo, C.; Charnet, C.; Spinnewyn, B.; Cornet, S.; Rolland, A.; Marin, J.G.; Bigg, D.; Chabrier, P.E. Phenolic thiazoles as novel orally-active neuroprotective agents. Bioorg. Med. Chem. Lett. 2004, 14, 157–160. [Google Scholar] [CrossRef]
  18. Patt, W.C.; Hamilton, H.W.; Taylor, M.D. Structure-activity relationships of a series of 2-amino-4-thiazole-containing rennin inhibitors. Med. Chem. 1992, 35, 2562–2572. [Google Scholar] [CrossRef]
  19. Kearney, P.C.; Fernandez, M.; Flygare, J.A. Solid-phase synthesis of 2-aminothiazoles. J. Org. Chem. 1998, 63, 196–200. [Google Scholar] [CrossRef]
  20. Haddad, N.; Tan, J.; Farina, V. Convergent synthesis of the quinolone substructure of BILN 2061 via carbonylative sonogashira coupling/cyclization. J. Org. Chem. 2006, 71, 5031–5034. [Google Scholar] [CrossRef]
  21. Venkatachalam, T.K.; Qazi, S.; Samuel, P.; Uckun, F.M. Substituted heterocyclic thiourea compounds as a new class of anti-allergic agents inhibiting IgE/FCeRI receptor mediated mast cell leukotriene release. Bioorg. Med. Chem. 2003, 11, 1095–1105. [Google Scholar] [CrossRef]
  22. Hargrave, K.D.; Hess, F.K.; Oliver, J.T. N-(4-Substituted-thiazolyl)oxamic acid derivatives, a new series of potent, orally active antiallergy agents. J. Med. Chem. 1983, 26, 1158–1163. [Google Scholar] [CrossRef]
  23. Venkatachalam, T.K.; Mao, C.; Uckun, F.M. Effect of stereochemistry on the anti-HIV activity of chiral thiourea compounds. Bioorg. Med. Chem. 2004, 12, 4275–4284. [Google Scholar] [CrossRef]
  24. Bell, F.W.; Cantrell, A.S.; Hogberg, M.; Jaskunas, S.R.; Johanson, N.G.; Zhou, X. Phenethylthiazoletthiourea (PETT) compounds, a new class of HIV-1 reverse transcriptase inhibitors. 1. Synthesis and basic structure-activity relationship studies of PETT analogs. J. Med. Chem. 1995, 38, 4929–4936. [Google Scholar] [CrossRef]
  25. Barf, T.; Vallgarda, J.; Emond, R.; Ohman, B.; Alberts, P.; Abrahmsen, L. Arylsulfonamidothiazoles as a new class of potencial antidiabetic drugs, discovery of potent and selective inhibitors of the 11β-hydroxysteroid dehydrogenase type 1. J. Med. Chem. Lett. 2002, 45, 3813–3815. [Google Scholar]
  26. Guannessi, F.; Chiodi, P.; Marzi, M.; Minetti, P.; Pessotto, P.; Tinti, M.; Carminati, P.; Arduini, A. Reversible carnitine palmitoyltransferase inhibitors with broad chemical diversity as potencial antidiabetic agents. J. Med. Chem. 2001, 44, 2383–2386. [Google Scholar] [CrossRef]
  27. Bienayme, H.; Bouzid, K. A new heterocyclic multicomponent reaction for the combinatorial synthesis of fused 3-aminoimidazoles. Angew. Chem. Int. Ed. 1998, 37, 2234–2237. [Google Scholar] [CrossRef]
  28. Maradiya, H.; Patel, V.S. Synthesis and dyeing performance of some novel heterocyclic azo disperse dyes. J. Braz. Chem. Soc. 2001, 12, 710–714. [Google Scholar]
  29. Hu, D.; Liu, S.; Huang, T.; Yang, H.; Zhang, A. Synthesis and herbicidal activities of novel 4-(4-(5-methyl-3-arylisoxazol-4-yl)thiazol-2-yl)piperidyl carboxamides and thiocarboxyamides. Molecules 2009, 14, 1288–1303. [Google Scholar] [CrossRef]
  30. Geronikaki, A.; Hadjipavlou-Litina, D.; Chatziopoulos, C.; Soloupis, G. Synthesis and biological evaluation of 4,5-disubstituted-thiazolyl amides, derivatives of 4-hydroxy-piperidine or of 4-N-methylpiperazine. Molecules 2003, 8, 472–479. [Google Scholar] [CrossRef]
  31. Vaickelioniene, R.; Mickevicius, V. Cyclization of N-fluorophenyl-β-alanines and properties of the obtained products. Chem. Heterocycl. Compd. 2006, 6, 862–869. [Google Scholar]
  32. Mickevicius, V.; Patupaite, A. Cyclization of N-(2,4,6-trimethylphenyl)-β-alanines. Chem. Heterocycl. Compd. 2000, 7, 951–954. [Google Scholar]
  33. Vaickelionienė, R.; Mickevičius, V.; Mikulskiene, G. Synthesis and cyclizations of N-(2,3-,3,4- and 3,5-dimethylphenyl)-β-alanines. Molecules 2005, 10, 407–416. [Google Scholar] [CrossRef]
  34. Blėkaitytė, V.; Jonuškienė, I.; Mickevičius, V. The influence of n-substituted β-alanines containing naphthoquinone and thiazole moieties on the growth of st. John’s wort (Hypericum perforatum L.) and its ability to accumulate metabolites. Chem. Tech. 2012, 4, 29–35. [Google Scholar]
  35. Chowdhry, M.M.; Mingos, D.M.P.; White, A.J.P.; Williams, D.J. Syntheses and characterization of 5-substituted hydantoins and thiazolines—Implications for crystal engineering of hydrogen bonded assemblies. Crystal structures of 5-(2-pyridylmethylene)-hydantoin, 5-(2-pyridylmethylene)-2-thiohydantoin, 5-(2-pyridylmethylene)thiazolidine-2,4-dione, 5-(2-pyridylmethylene)rhodanine and 5-(2-pyridylmethylene)pseudothiohydantoin. Chem. Soc. Perkin Trans. 2000, 1, 3495–3504. [Google Scholar]
  36. El-Najjar, N.; Gali-Muhtasib, H.; Ketota, R.A.; Vuorela, P.; Urtti, A.; Vuorela, H. The chemical and biological activities of quinones: Overview and implications in analytical detection. Phytochem. Rev. 2011, 10353–103370. [Google Scholar]
  37. Burg, M.B.; Ferraris, J.D. Intracellular organic osmolytes: Function and regulation. J. Biol. Chem. 2008, 283, 7309–7313. [Google Scholar] [CrossRef]
  38. Stasevych, M.; Lubenets, V.; Musyanovych, R.; Novikov, V.; Mickevičius, V.; Beresnevičius, Z.J.; Rutkauskas, K. Novel thiazolone derivatives of N-aryl-β-alanines. Chem. Heterocycl. Compd. 2011, 47, 1050–1052. [Google Scholar] [CrossRef]
  39. Franklin, A.; Acar, J.; Anthony, F.; Gupta, R.; Nicholls, T.; Tamura, Y.; Thompson, S.; Thelfall, E.J.; Vose, D.; van Vuuren, M.; et al. Antimicrobial resistance: Harmonisation of national antimicrobial resistance monitoring and surveillance programmes in animals and in animal-derived food. Rev. Sci. Tech. Off. Int. Epiz. 2001, 20, 859–870. [Google Scholar]
  40. Goodson, B.; Ehrhardt, A.; Ng, S.; Nuss, J.; Johnson, K.; Giedlin, M.; Yamamoto, R.; Krebber, A.; Ladner, M.; Giacona, M.B.; et al. Characterization of novel antimicrobial peptoids. Antimicrob. Agents Chemother. 1999, 43, 1429–1434. [Google Scholar]
  41. Petersen, A.; Aarestrup, F.M.; Hofshagen, M.; Sipilä, H.; Franklin, A.; Gunnarsson, E. Harmonization of Antimicrobial Susceptibility Testing among Veterinary Diagnostic Laboratories in the Five Nordic Countries. Microb. Drug Resist. 2003, 9, 381–388. [Google Scholar] [CrossRef]
  • Sample Availability: Samples of the compounds 114 are available from the authors.
Back to TopTop