Interaction of Pyrimidine Derivatives with Human Serum Albumin†

doi:10.7503/cjcu20170063

Abstract

Abstract:

The interaction of eight pyrimidine derivatives(PDs, 5-FU marked as A is the positive control, other seven samples have been made in our laboratory and numbered from B to H, respectively) with human serum albumin(HSA) have been investigated by means of steady state fluorescence, laser flash photolysis(LFP), UV-Vis absorption spectroscopy and molecular docking techniques under simulative physiological coditions. The fluorescence quenching mechanism of HSA and different pyrimidine derivatives have been analyzed by the Stern-Volmer equation and laser flash photolysis techniques. From these results, we can know that pyrimidine derivatives numbered from A to E are static quenching, but pyrimidine derivatives numbered from G to H are dynamic quenching. Depending on double reciprocal curve, we can obtain the binding constants(K_a) and the number of binding sites(n) of the five pyrimidine derivatives of static quenching and HSA. These data show that the protein and pyrimidine derivatives combinate with weak binding force and the number of binding site are equal to one. Fitting the thermodynamic parameters ΔH, ΔS and ΔG can faciliate to understand the binding force types for pyrimidine derivatives and HSA, and pyrimidine derivative B, C and E are the electrostatic and hydrophobic interactions, while for pyrimidine derivatives A and D are hydrogen bond and van der Waals force, and these experimental results are consistent with molecular docking results. According to non-radiative energy transfer theory(FRET), the binding distance(r) between HSA and pyrimidine derivaties can be obtained, which are less than 4 nm, and these results are conform to the energy transfer theory. With synchronous fluorescence, three-dimensional fluorescence and circular dichroism spectroscopy, the space conformation changes of HSA in the drugs combining process can be investigated. The results show that pyrimidine derivatives B, D and E have no influence on the secondary and tertiary structure of HSA. pyrimidine derivative A and C have no effect on secondary structure of HSA, but the hydrophobicity around the aromatic amino acids for the tertiary structure are slightly enhanced. Therefore, HSA can be used as an excellent carrier to transport and store these five pyrimidine derivatives.

Key words: Human serum albumin(HSA), Pyrimidine derivative, Three dimensional fluorescence technique, Laser flash photolysis, Molecular docking technique

CLC Number:

O623
Q657.3

TrendMD:

TANG Qian, SU Jinhong, CAO Hongyu, WANG Lihao, SHI Fei, WANG Ailing, GONG Tingting, JIN Xiaojun, ZHENG Xuefang. Interaction of Pyrimidine Derivatives with Human Serum Albumin^†[J]. Chem. J. Chinese Universities, 2017, 38(11): 1982.

Figures/Tables 15

References 43

[1]	Kratochwil N. A., Huber W., Muller F., Kansy M., Gerber P. R., Biochem. Pharmacol., 2002, 64(9), 1355-1374
[2]	Ahmad B., Parveen S., Khan R. H., Biomacromolecules,2006, 7(4), 1350-1356
[3]	Kelarakis A., Castelletto V., Krysmann M. J., Havredaki V., Viras K., Hamley I. W., Biomacromolecules,2008, 9(5), 1366-1371
[4]	Ruiz J., Vicente C., de Haro C., Bautista D., Inorg. Chem., 2013, 52( 2), 974-982
[5]	Han Y. C., Fan Y. R., Huang X., Wang Y. L., Chem. J. Chinese Universities,2010, 31(6), 1014-1023
	(韩玉淳, 樊艳茹, 黄旭, 王毅琳. 中国科学: 化学, 2014, 44(6), 1014-1023)
[6]	Wen J. Y., Lü B. B., Zhang Y., Wang J. M., Ying X., Wang H., Ji L. N., Liu H. Y., Chem. J. Chinese Universities,2010, 31(6), 1033-1041
	(闻金燕, 吕标彪, 张阳, 王家敏, 应晓, 王慧, 计亮年, 刘海洋. 高等学校化学学报, 2015, 36(6), 1033-1041)
[7]	Wang C., Wu Q. H., Wang Z., Zhao J., Anal. Sci., 2006, 22(3), 435-438
[8]	Maciᶏzek-Jurczyk M., Sułkowska A., Bojko B., Równicka-Zubik J., Szkudlarek-Hasnik A., Zubik-Skupień I., Góra A., Dubas M., Korzonek-Szlacheta I., Wielkoszyński T., Zurawiński W., Sosada K., Spectrochim. Acta A Mol. Biomol. Spectrosc., 2012, 89(4), 270-275
[9]	Maciᶏzek-Jurczyk M., Sułkowska A., Bojko B., Równicka-Zubik J., Sułkowski W. W., Spectrochim. Acta A Mol. Biomol. Spectrosc., 2011, 82(1), 189-190
[10]	Gomha S. M., Zaki Y. H., Abdelhamid A. O., Molecules,2015, 20(12), 21826-21839
[11]	Ghoneim K. M., El-Telbany F., Youssef K. M., J. Indian Chem. Soc., 1986, 63, 914-917
[12]	Vishwas D., Suryawanshi Prashant V. A., Anil H. G., Shivajirao R. P., Govind B. K., J. Photochem. Photobiol. B: Biology,2013, 118, 1-8
[13]	Zhao P. L., You W. W., Duan A. N., Chem. J. Chinese Universities,2010, 31(5), 580-587
	(赵培亮, 游文玮, 段安娜. 药学学报, 2012, 47(5), 580-587)
[14]	Yasumoto M., Yamawaki I., Marunaka T., Hashimoto S., J. Med. Chem., 1978, 21(8), 738-741
[15]	Masao T. A. D. A., Bull. Chem. Soc. Japan, 1975, 48(11), 3427-3428
[16]	Mohammad H., RezaY., Asghar T. K., Farhad P., AliKhalafi N., Chem. J. Chinese Universities,2010, 31, 374-379
[17]	Arica B., Calis S., Kas H., Sargon M., Hincal A., Int. J. Pharm., 2002, 242(1/2), 267-269
[18]	Wang X. D., Chi G. C., Chen R. Y., Chin. J. Synth. Chem., 2001, 9(1), 1-2
[19]	Rodgers K. R., Curr. Opin. Chem. Biol., 1999, 3(2), 158-167
[20]	Seetharamappa J., Kamat B. P., Chem. Pharm. Bull., 2004, 52, 1053-1057
[21]	Yang Y., Hu Q., Fan Y., Shen H., Chem. J. Chinese Universities,2010, 31(2), 432-436
[22]	Kandagal P. B., Seetharamappa J., Ashoka S., Shaikh S. M., Manjunatha D. H., Int. J. Biol. Macromol., 2006, 39(4/5), 234-239
[23]	Ray D., Paul B. K., Guchhait N., Chem. J. Chinese Universities,2010, 31, 18-27
[24]	Sharma A. S., Ilanchelian M., Chem. J. Chinese Universities,2010, 31(30), 9461-9476
[25]	Wei P., Fei D., Jiang Y. T., Peng Y. K., J. Agric. Food Chem., 2014, 62, 2271-2283
[26]	Ganesh B. M., Shateesh B., Sudhir K., Santosh K. H., Chem. J. Chinese Universities,2010, 31, 20944-20950
[27]	Shahabadi N., Khorshidi A., Moghadam N. H., Chem. J. Chinese Universities,2010, 31, 627-632
[28]	Wang R., Wang X., Li Z., Xie Y., Yang L., Shi J., Chang J., Spectrochim Acta A: Mol. Biomol. Spectrosc., 2014, 132, 786-794
[29]	Ma J., Zheng X.F., Tang Q., Yang Y. J., Sun X., Gao D. B.,Chem. J. Chinese Universities, 2008, 29(2), 258-263
	(马静, 郑学仿, 唐乾, 杨彦杰, 孙霞, 高大彬. 高等学校化学学报, 2008, 29(2), 258-263)
[30]	Wang Y. Q., Zhang H. M., Zhang G. C., Liu S. X., Zhou Q. H., Fei Z. H., Liu Z. T., Int. J. Biol. Macromol., 2007, 41(3), 243-250
[31]	Ross P. D., Subramanian S., Biochemistry,1981, 20(11), 3096-3102
[32]	Bourassa P., Dubeau S., Maharvi G. M., Fauq A. H., Thomas T. J., Tajmir-Riahi H. A., Eur. J. Med. Chem., 2011, 46(9), 4344-4353
[33]	Molina-Bolívar J. A., Galisteo-González C. R. F., Medina-O’ D. M., Parra A., J. Mol. Liq., 2015, 208, 304-313
[34]	Xie X., Lü W., Chen X., J. Hazard. Mater., 2013, 248-249, 347-354
[35]	Han X. L., Tian F. F., Ge Y. S, Jiang F. L., Lai L., Li D. W., Yu Q. L., Wang J., Lin C., Liu Y., Int. J. Biol. Macromol., 2012, 109, 1-11
[36]	Li X., Li C., Jiang J. H., Gu H. W., Wei D. L., YE L. J., Hu J. L., Xiao S. X., Zhang H., Li X., Li Q. G., Chem. J. Chinese Universities,2010, 31(2), 166-171
[37]	Maryam S., Hassan M. T., Ali Akbar S., J. Lumin., 2015, 167, 391-398
[38]	Shen H., Gu Z., Jian K., Qi J., J. Pharm. Biomed. Anal., 2013, 75, 86-93
[39]	Zhang J. L., Gu H. M., Zhang X. H., Carbohyd. Res., 2014, 384, 102-111
[40]	Tu B., Wang Y., Mi R., Ouyang Y., Hu Y. J., Spectrochim Acta A: Mol. Biomol. Spectrosc., 2015, 149, 536-543
[41]	Ge Y. S., Tai S. X., Xu Z. Q., Lai L., Tian F. F., Li D. W., Jiang F. L., Liu Y., Gao Z. N., Langmuir,2012, 28, 5913-5920
[42]	Cheng H., Liu H., Zhang Y., J. Lumin., 2009, 129, 1196-1203
[43]	Nicolott O., Catto M., Giangreco I., Eur. J. Med. Chem., 2012, 58, 368-376

PDs	IC₅₀/(μmol·L^-1)		PDs	IC₅₀/(μmol·L^-1)
PDs	HCC(7721)		MC(A375)	HCC(7721)	MC(A375)
A	23.65	9.42	E	166.90	28.94
B	88.05	81.37	F	22.43	12.85
C	9.53	17.86	G	23.57	27.28
D	16.66	34.76	H	12.22	48.19

PDs	IC₅₀/(μmol·L^-1)		PDs	IC₅₀/(μmol·L^-1)
PDs	HCC(7721)		MC(A375)	HCC(7721)	MC(A375)
A	23.65	9.42	E	166.90	28.94
B	88.05	81.37	F	22.43	12.85
C	9.53	17.86	G	23.57	27.28
D	16.66	34.76	H	12.22	48.19

PDs	T/K	K_SV/ (L·mol^-1)	K_q/ (L·mol^-1·s^-1)	Quenching mechanism	PDs	T/K	K_SV/ (L·mol^-1)	K_q/ (L·mol^-1·s^-1)	Quenching mechanism
A	300	3.206×10⁴	3.20×10¹²	Static quenching	E	300	3.32×10³	3.32×10¹¹	Static quenching
	310	3.17×10⁴	3.17×10¹²			310	6.80×10³	6.80×10¹¹
	320	3.05×10³	3.05×10¹²			320	6.12×10³	6.12×10¹¹
B	300	7.05×10³	7.05×10¹¹	Static quenching	F	300	5.01×10³	5.01×10¹¹	Dynamic quenching
	310	6.82×10³	6.82×10¹¹			310	3.32×10³	3.32×10¹¹
	320	6.36×10³	6.36×10¹¹			320	3.69×10³	3.69×10¹¹
C	300	4.73×10⁴	4.73×10¹²	Static quenching	G	300	4.91×10³	4.91×10¹¹	Dynamic quenching
	310	4.34×10⁴	4.34×10¹²			310	5.65×10³	5.65×10¹¹
	320	3.71×10⁴	3.71×10¹²			320	6.60×10³	6.60×10¹¹
D	300	4.76×10³	4.76×10¹¹	Static quenching	H	300	4.27×10³	4.27×10¹¹	Dynamic quenching
	310	4.40×10³	4.40×10¹¹			310	4.42×10³	4.42×10¹¹
	320	4.20×10³	4.20×10¹¹			320	4.72×10³	4.72×10¹¹

PDs	T/K	K_SV/ (L·mol^-1)	K_q/ (L·mol^-1·s^-1)	Quenching mechanism	PDs	T/K	K_SV/ (L·mol^-1)	K_q/ (L·mol^-1·s^-1)	Quenching mechanism
A	300	3.206×10⁴	3.20×10¹²	Static quenching	E	300	3.32×10³	3.32×10¹¹	Static quenching
	310	3.17×10⁴	3.17×10¹²			310	6.80×10³	6.80×10¹¹
	320	3.05×10³	3.05×10¹²			320	6.12×10³	6.12×10¹¹
B	300	7.05×10³	7.05×10¹¹	Static quenching	F	300	5.01×10³	5.01×10¹¹	Dynamic quenching
	310	6.82×10³	6.82×10¹¹			310	3.32×10³	3.32×10¹¹
	320	6.36×10³	6.36×10¹¹			320	3.69×10³	3.69×10¹¹
C	300	4.73×10⁴	4.73×10¹²	Static quenching	G	300	4.91×10³	4.91×10¹¹	Dynamic quenching
	310	4.34×10⁴	4.34×10¹²			310	5.65×10³	5.65×10¹¹
	320	3.71×10⁴	3.71×10¹²			320	6.60×10³	6.60×10¹¹
D	300	4.76×10³	4.76×10¹¹	Static quenching	H	300	4.27×10³	4.27×10¹¹	Dynamic quenching
	310	4.40×10³	4.40×10¹¹			310	4.42×10³	4.42×10¹¹
	320	4.20×10³	4.20×10¹¹			320	4.72×10³	4.72×10¹¹

Sample	τ₁/ns	τ₂/ns	B₁	B₂	τ/ns	χ²	k₂	C₂/(mol·L^-1)
HSA	2.24	6.62	-0.30	0.81	7.25	1.13	1.51×10⁵	1.59×10^-6
HSA+A	2.18	6.61	-0.37	0.95	7.26	1.25	1.51×10⁵	1.64×10^-6
HSA+B	2.22	6.52	-0.38	0.97	7.18	0.97	1.53×10⁵	1.64×10^-6
HSA+C	2.06	5.58	-0.24	0.59	6.20	1.02	1.79×10⁵	1.69×10^-6
HSA+D	2.26	6.62	-0.39	0.96	7.32	1.26	1.51×10⁵	1.68×10^-6
HSA+E	1.92	5.92	-0.26	0.72	6.45	0.69	1.69×10⁵	1.57×10^-6
HSA+F	1.89	6.00	-0.36	1.05	6.50	1.69	1.67×10⁵	1.52×10^-6
HSA+G	2.00	5.83	-0.43	1.07	6.44	1.25	1.72×10⁵	1.67×10^-6
HSA+H	2.05	6.09	-0.52	1.29	6.72	1.80	1.64×10⁵	1.68×10^-6