Interaction of 2-{[4-Amino-5-(3,4,5-trimethoxy-benzyl)-pyrimidin-2-ylimino]-methyl}-6-methoxy-phenol with S. pombe Cells and BSA†

doi:10.7503/cjcu20130797

Abstract

Abstract:

A new compound 2-{[4-amino-5-(3,4,5-trimethoxy-benzyl)-pyrimidin-2-ylimino]-methyl}-6-methoxy-phenol(APYP) was synthesized using equivalent moles of o-vanillin(C₈H₈O₃) and trimethoprim(C₁₄H₁₈N₄O₃). Elemental analysis, molar conductivity, UV-Visual, IR and NMR spectroscopy were employed to characterize the composition and structure of APYP. The analytical results showed that the molecular formula of the compound is C₂₂H₂₄N₄O₅. The thermokinetic properties of the action of APYP on growth meta-bolism of S. pombe were studied by a TAM air isothermal calorimeter at 301.15 K. The microbial growth rate constant(k), inhibition ratio(I) and half inhibition concentration(IC₅₀) were obtained. Experimental results show that the k values of S. pombe decreased and I values of S. pombe increased with the increase in the concentrations of APYP and the IC₅₀ value of APYP was found to be 123.20 mg/L. The APYP has good inhibition activity on the growth of S. pombe. Besides, fluorescence spectroscopy and UV-Vis absorption spectroscopy were employed to investigate the interactions of APYP with bovine serum albumin(BSA) under the approximatively physiological conditions. The binding constants, binding sites, binding distance, and the thermodynamic parameters were calculated. The energy transfer between the donor(BSA) and acceptor(APYP) was investigated. Furthermore, synchronous fluorescence spectroscopy was used to investigate the structural change of BSA molecules with additions of APYP.

Key words: o-Vanillin, Trimethoprim, Thermokinetic property, Bovine serum albumin, S. pombe cell

CLC Number:

O641
O633.12

TrendMD:

JIANG Jianhong, LI Xu, XIAO Shengxiong, GU Huiwen, LI Chuanhua, YANG Ping, WEI Deliang, HE Dugui, LI Aitao, LI Xia, YAO Feihong, LI Qiangguo. Interaction of 2-{[4-Amino-5-(3,4,5-trimethoxy-benzyl)-pyrimidin-2-ylimino]-methyl}-6-methoxy-phenol with S. pombe Cells and BSA^†[J]. Chem. J. Chinese Universities, 2014, 35(4): 831.

Figures/Tables 12

Fig.1 Power-time curves for the growth of S. pombe affected by APYPConcentrations of APYP/(mg·L-1): a. 0; b. 40; c. 60; d. 80; e. 100; f. 120; g. 140.

Table 1 Thermokinetic parameters of the growth of S.pombe affected by different concentrations of APYP at 301.15 K

c(APYP)/(mg·L^-1)	k^*/s^-1	R	I	IC₅₀/(mg·L^-1)
0	7.20×10^-5±5×10^-8	0.9994	0
40	6.26×10^-5±4×10^-8	0.9992	13.1
60	5.37×10^-5±4×10^-8	0.9993	25.4
80	4.76×10^-5±2×10^-8	0.9998	33.9	123.2
100	4.10×10^-5±7×10^-9	1.0000	43.1
120	3.63×10^-5±1×10^-8	0.9995	49.6
140	3.40×10^-5±2×10^-8	0.9991	52.8

Fig.2 Rate constants of growth(k) of S. pombe with concentration of APYP(c)

Fig.3 Inhibitive ratio(I) of S. pombe with concentration of APYP(c)

Fig.4 Quenching effects of APYP on BSA fluore-scence intensity at 298.15 Kλex=280 nm, λem=340 nm, c(BSA)=2.0×10-6 mol/L, c(APYP)=2.0×10-6 mol/L. c(APYP)/c(BSA), a—k: 0, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0.

Fig.5 Stem-Volmer plots for the fluorescence quenching of BSA by APYP at different temperaturesa. 298.15 K, slope=1.0622, R=0.9996; b. 302.15 K, slope=1.0194, R=0.9985; c. 306.15 K, slope=0.9414, R=0.9987; d. 310.15 K, slope=0.9133, R=0.9977.

Fig.6 Lineweaver-Burk plots for the fluorescence quenching of BSA by APYP at different temperaturesa. 298.15 K, slope=0.00277, R=0.9976; b. 302.15 K, slope=0.00301, R=0.9875; c. 306.15 K, slope=0.00379, R=0.9962; d. 310.15 K, slope=0.00425, R=0.09960.

Table 2 Relative thermodynamic parameters of APYP-BSA system

Temperature/K	K_LB	$Δ H 0 —$ /(kJ·mol^-1)	$Δ G 0 —$ /(kJ·mol^-1)	$Δ S 0 —$ /(J·mol^-1·K^-1)
298.15	1.317×10⁶	-22.997	-34.929	40.020
302.15	1.241×10⁶	-22.997	-35.248	40.546
306.15	1.013×10⁶	-22.997	-35.198	39.853
310.15	9.198×10⁵	-22.997	-35.409	40.019

Table 2 Relative thermodynamic parameters of APYP-BSA system

Temperature/K	K_LB	$Δ H 0 —$ /(kJ·mol^-1)	$Δ G 0 —$ /(kJ·mol^-1)	$Δ S 0 —$ /(J·mol^-1·K^-1)
298.15	1.317×10⁶	-22.997	-34.929	40.020
302.15	1.241×10⁶	-22.997	-35.248	40.546
306.15	1.013×10⁶	-22.997	-35.198	39.853
310.15	9.198×10⁵	-22.997	-35.409	40.019

Fig.7 Double-log plots of APYP quenching effect on BSA fluorescence at different temperatures

Table 3 Binding constants and number of binding sites of APYP-BSA system

Temperature/K	K_A/(L·mol^-1)	n	R	Temperature/K	K_A/(L·mol^-1)	n	R
298.15	3.3´10⁵	0.9026(≈1)	0.9968	306.15	7.5´10⁵	0.9820(≈1)	0.9980
302.15	5.6´10⁵	0.9521(≈1)	0.9951	310.15	1.0´10⁶	1.0139(≈1)	0.9971

Fig.8 Spectral overlap between the absorption spectrum of APYP(a) and fluorescence emission spectrum of BSA(b) at 298.15 Kc(BSA)=2.0×10-6 mol/L; c(APYP)=2.0×10-6, 5 mmol/L. Tris-HCl buffer, pH=7.4. Overlapped region is highlighted.

Fig.9 Synchronous fluorescence spectra of BSA at 298.15 K(A) Δλ=15 nm; (B) Δλ=60 nm. c(BSA)=c(APYP)=2.0×10-6 mol/L. c(APYP)/c(BSA), a—e: 0, 1.0, 2.0, 4.0, 5.0.

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