Preparation of the Protein/Polyphenylboronic Acid Nanospheres for Drug Loading and Unloading

doi:10.6023/A22110464

Abstract

Protein nanoparticles (NPs), biocompatible and biodegradable, can be easily surface modified. In particular, amphiphilic proteins act as “surfactants” that help form microparticles while undergoing molecular chain rearrangement. These NPs have been successfully used as drug delivery systems, improving bioavailability and reducing the toxic effects of drug molecules. The use of regenerated silk protein (RSF) with m-acrylamidophenylboronic acid (APBA) composites as drug carriers for loading anti-inflammatory herbal extracts was reported. Firstly, a simple and rapid method was used to prepare silk protein/polyphenylboronic acid nanospheres, in brief, RSF solution and a certain amount of initiator were added to APBA solution, and the pH was adjusted by NaOH, and the polymerization was initiated by heating at 90 ℃ under nitrogen protection with stirring at 500 r/min. After 2 h of reaction, a milky solution was obtained, which formed silk protein/benzeneboronic acid nanospheres with hydrophobic interior and hydrophilic surface. The drug-loaded silk protein/polyphenylboronic acid nanospheres with an average size of 550 to 600 nm were prepared by mixing with the drug solution after dialysis and stirring at room temperature for 12 h to load the drug by adsorption. By the same method, drug-loaded albumin/polyphenylboronic acid microspheres and collagen/polyphenylboronic acid microspheres with sizes around 260 nm and 100 nm, respectively, could be prepared. The results observed by scanning and projection electron microscopy and dynamic light scattering showed that the drug-loaded silk protein/polyphenylboronic acid nanomicrospheres displayed regular spherical shape indicating smoothness and good dispersion with no obvious aggregation. The highest drug loading rate was about 13.4%, and the encapsulation rate was over 90%. Also, such drug-loaded nanospheres could achieve controlled release for about seven days and their slow release behavior was pH-responsive, with faster drug release in buffer solution at pH=5.5 than in buffer solution at pH=7.4. In addition, the synergistic interaction of the silk protein/polyphenylboronic acid nanomicrospheres with the subject drug improved its free radical scavenging rate and scavenging efficiency, which was superior to that of the direct drug delivery group. Thus, three protein/polyphenylboronic acid nanomicrospheres with different sizes, electrical properties and drug release rates may be adaptable to a wide range of intravenous and subcutaneous or intraperitoneal drug delivery needs and have great potential for clinical applications.

Key words: proteins, phenylboronic acid, anti-inflammatory drugs, nanocarriers, controlled release

Cite this article

Xueyang Yin, Kai Gu, Zhengzhong Shao. Preparation of the Protein/Polyphenylboronic Acid Nanospheres for Drug Loading and Unloading[J]. Acta Chimica Sinica, 2023, 81(2): 116-123.

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Fig. & Tab. 6

Figure 1. SEM and TEM images of RSF-PAPBA (a, b), Pin@RSF-PAPBA (c, d), Far@RSF-PAPBA (e, f), and Dio@RSF-PAPBA (g, h) nanospheres

Sample	Hydrodynamic diameter/nm	PDI	Zeta potential/mV
RSF-PAPBA	540	0.04	–23.0
Pin@RSF-PAPBA	590	0.06	–31.0
Far@RSF-PAPBA	560	0.09	–32.5
Dio@RSF-PAPBA	570	0.07	–30.2
RSF	540	0.12	–23.3
Far@RSF	290	0.18	–34.4

Table 1. Hydrodynamic diameter (nm), polydispersity index (PDI) and Zeta potential (mV) of RSF nanospheres before or after drug loading

Figure 2. SEM images of BSA-PAPBA (a), Far@BSA-PAPBA (b), GEL-PAPBA (c) and Far@GEL-PAPBA (d) nanospheres

Sample	Hydrodynamic diameter/nm	PDI	Zeta potential/mV
BSA-PAPBA	260	0.05	–24.2
Far@BSA-PAPBA	290	0.09	–46.2
GEL-PAPBA	170	0.04	+21.5
Far@GEL-PAPBA	180	0.06	+14.3

Table 2. Hydrodynamic diameter (nm), polydispersity index (PDI) and Zeta potential (mV) of BSA-PAPBA or GEL-PAPBA nanospheres before and after drug loading

Figure 3. Release profiles of drug loaded protein/polyphenylboronic acid nanospheres in phosphate buffer solution. (a) Release profiles of different microspheres loaded with farrerol at pH=5.5; (b) release profiles of RSF-PAPBA microspheres loaded with azathioprine at different pH; (c, d) release profiles of drug loaded RSF-PAPBA for three different drugs, namely, pinocembrin, farrerol and diosmin, at pH=7.4 and pH=5.5

Figure 4. The photographs (a, c, e) and quantitative curves (b, d, f) of the scavenging effects regarding DPPH radical of the corresponded drug and its RSF-PAPBA loaded nanospheres within 30 min

References 47

[1]	Pahwa, R.; Goyal, A.; Jialal, I. Chronic Inflammation. Vol. 1, Ed.: Bansal, P., StatPearls, Petersburg, 2020, p. 7.
[2]	Luo, W. X.; Wen, G.; Yang, L.; Tang, J.; Wang, J. G.; Wang, J. H.; Zhang, S. Y.; Zhang, L.; Ma, F.; Xiao, L. L.; Wang, Y.; Li, Y. Theranostics. 2017, 7, 452. doi: 10.7150/thno.16677
[3]	Kamimura, M.; Furukawa, T.; Akiyama, S.; Nagasaki, Y. Biomater. Sci. 2013, 1, 361. doi: 10.1039/c2bm00156j
[4]	Davis, M. E.; Brewster, M. E. Nat. Rev. Drug Discov. 2004, 3, 1023. doi: 10.1038/nrd1576 pmid: 15573101
[5]	Lee, C. C.; MacKay, J. A.; Frechet, J. M. J.; Szoka, F. C. Nat. Biotechnol. 2005, 23, 1517. doi: 10.1038/nbt1171
[6]	Serajuddin, A. T. M. Adv. Drug Deliver. Rev. 2007, 59, 603. doi: 10.1016/j.addr.2007.05.010 pmid: 17619064
[7]	Akers, M. J. J. Pharm. Sci. 2002, 91, 2283. doi: 10.1002/jps.10154
[8]	Gregoriadis, G.; Wills, E. J.; Swain, C. P.; Tavill, A. S. Lancet. 1974, 303, 1313. doi: 10.1016/S0140-6736(74)90682-5
[9]	Juliano, R. L. Prog. Clin. Biol. Res. 1976, 9, 21. pmid: 1030801
[10]	Kobayashi, H.; Turkbey, B.; Watanabe, R.; Choyke, P. L. Bioconjugate Chem. 2014, 25, 2093. doi: 10.1021/bc500481x pmid: 25385142
[11]	Mitchell, M. J.; Jain, R. K.; Langer, R. Nat. Rev. Cancer. 2017, 17, 659. doi: 10.1038/nrc.2017.83 pmid: 29026204
[12]	Zheng, P.; Yang, L.; Chen, J. J.; Xu, W. G.; Li, G.; Ding, J. X. Chinese Chem. Lett. 2020, 31, 1178. doi: 10.1016/j.cclet.2019.12.001
[13]	Kosheli, T. M.; George, F. B.; Li, X. T.; Chen, Z. J.; He, W. Chinese Chem. Lett. 2022, 33, 587. doi: 10.1016/j.cclet.2021.08.020
[14]	Saha, A.; Pradhan, N.; Chatterjee, S.; Singh, R. K.; Trivedi, V.; Bhattacharyya, A.; Manna, D. ACS Appl. Nano Mater. 2019, 2, 3671. doi: 10.1021/acsanm.9b00607
[15]	Xie, M.; Fan, D.; Chen, Y.; Zhao, Z.; He, X.; Li, G.; Lan, P. Biomaterials. 2016, 103, 33. doi: 10.1016/j.biomaterials.2016.06.049
[16]	Wang, S.; Xu, T.; Yang, Y.; Shao, Z. ACS Appl. Mater. Interfaces. 2015, 7, 21254. doi: 10.1021/acsami.5b05335
[17]	Shao, Z. Z.; Vollrath, F. Nature. 2002, 418, 741. doi: 10.1038/418741a
[18]	Wu, M.; Yang, W.; Chen, S.; Yao, J.; Shao, Z.; Chen, X. J. Mater. Chem. B. 2018, 6, 1179. doi: 10.1039/C7TB03113K
[19]	Sun, N.; Lei, R.; Xu, J. H.; Kundu, S. C.; Cai, Y. R.; Yao, J. M.; Ni, Q. Q. J. Mater. Sci. 2019, 54, 3319. doi: 10.1007/s10853-018-3022-9
[20]	Yang, W. H.; Yu, S. Y.; Chen, S.; Liu, Y. Z.; Shao, Z. Z.; Chen, X. Acta Chim. Sinica. 2014, 72, 1164. (in Chinese) doi: 10.6023/A14080596
	(杨文华, 俞淑英, 陈胜, 刘也卓, 邵正中, 陈新, 化学学报, 2014, 72, 1164.) doi: 10.6023/A14080596
[21]	Wang, S. H.; Sun, L. N.; Cao, H.; Zhong, Y. M.; Shao, Z. Z. Acta Chim. Sinica. 2021, 79, 1023. (in Chinese) doi: 10.6023/A21050203
	(王苏杭, 孙灵娜, 曹涵, 钟一鸣, 邵正中, 化学学报, 2021, 79, 1023.) doi: 10.6023/A21050203
[22]	Huang, H.; Yang, D. P.; Liu, M. H.; Wang, X. S.; Zhang, Z. Y.; Zhou, G. D.; Liu, W.; Cao, Y. L.; Zhang, W. J.; Wang, X. S. Int. J. Nanomed. 2017, 12, 2829. doi: 10.2147/IJN.S128270 pmid: 28435261
[23]	Lu, H. X.; Noorani, L.; Jiang, Y. Y.; Du, A. W.; Stenzel, M. H. J. Mater. Chem. B. 2017, 5, 9591. doi: 10.1039/C7TB02902K
[24]	Yu, Z.; Yu, M.; Zhang, Z. B. Nanoscale Res. Lett. 2014, 9, 343. doi: 10.1186/1556-276X-9-343 pmid: 25114637
[25]	Azimi, B.; Nourpanah, P.; Rabiee, M.; Arbab, S. Iran Biomed. J. 2014, 18, 34.
[26]	Sahoo, N.; Sahoo, R. K.; Biswas, N.; Guha, A.; Kuotsu, K. Int. J. Biol. Macromol. 2015, 81, 317. doi: 10.1016/j.ijbiomac.2015.08.006
[27]	Kulkarni, S. A.; Feng, S. S. Pharm. Res. 2013, 30, 2512. doi: 10.1007/s11095-012-0958-3
[28]	Lee, B. R.; Ko, H. K.; Ryu, J. H.; Ahn, K. Y.; Lee, Y. H.; Oh, S. J.; Na, J. H.; Kim, T. W.; Byun, Y.; Kwon, I. C.; Kim, K.; Lee, J. Sci. Rep. 2016, 6, 35182. doi: 10.1038/srep35182
[29]	Ci, X.; Chu, X.; Wei, M.; Yang, X.; Cai, Q.; Deng, X. PLoS One. 2012, 7, e34634. doi: 10.1371/journal.pone.0034634
[30]	Fattori, V.; Rasquel-Oliveira, F. S.; Artero, N. A.; Ferraz, C. R.; Borghi, S. M.; Casagrande, R.; Verri, W. A. J. Nat. Prod. 2020, 83, 1018. doi: 10.1021/acs.jnatprod.9b00887 pmid: 32083866
[31]	Stephan, D. W. Acc. Chem. Res. 2015, 48, 306. doi: 10.1021/ar500375j
[32]	Chen, H.; Shao, Z. Z. Acta Polym. Sinica. 2018, 62, 987. (in Chinese)
	(陈红, 邵正中, 高分子学报, 2018, 62, 987.)
[33]	Chen, W.; Zhen, X.; Wu, W. Sci. China Chem. 2020, 63, 648. doi: 10.1007/s11426-019-9699-3
[34]	Wang, L. J.; Sheng, X. L.; Wang, J.; Zhang, Y. H. Chinese J. Org. Chem. 2021, 41, 567. (in Chinese) doi: 10.6023/cjoc202006060
	(王李娟, 盛显良, 王杰, 张玉辉, 有机化学, 2021, 41, 567.) doi: 10.6023/cjoc202006060
[35]	Cabral, H.; Matsumoto, Y.; Mizuno, K.; Chen, Q.; Murakami, M.; Kimura, M.; Kataoka, K. Nat. Nanotechnol. 2011, 6, 815. doi: 10.1038/nnano.2011.166 pmid: 22020122
[36]	Jiang, W.; Kim, B. Y.; Rutka, J. T.; Chan, W. C. Nat. Nanotechnol. 2008, 3, 145. doi: 10.1038/nnano.2008.30 pmid: 18654486
[37]	Reddy, S. T.; Van Der Vlies, A. J.; Simeoni, E.; Angeli, V.; Randolph, G. J.; O'Neil, C. P.; Hubbell, J. A. Nat. Biotechnol. 2007, 25, 1159. doi: 10.1038/nbt1332 pmid: 17873867
[38]	Wang, H.; Wang, S.; Su, H.; Chen, K. J.; Armijo, A. L.; Lin, W. Y.; Tseng, H. R. Angew. Chem. Int. Ed. 2009, 48, 4344. doi: 10.1002/anie.200900063 pmid: 19425037
[39]	Goodman, T. T.; Olive, P. L.; Pun, S. H. Int. J. Nanomed. 2007, 2, 265.
[40]	Tang, L.; Yang, X.; Dobrucki, L. W.; Chaudhury, I.; Yin, Q.; Yao, C.; Cheng, J. Angew. Chem. Int. Ed. 2012, 51, 12721. doi: 10.1002/anie.201205271 pmid: 23136130
[41]	Tang, L.; Yang, X.; Yin, Q.; Cai, K.; Wang, H.; Chaudhury, I.; Yao, C.; Zhou, Q.; Kwon, M.; Hartman, J. A.; Dobrucki, I. T.; Dobrucki, L. W.; Borst, L. B.; Lezmi, S.; Helferich, W. G.; Ferguson, A. L.; Fan, T. M.; Cheng, J. Proc. Natl. Acad. Sci. 2014, 111, 15344. doi: 10.1073/pnas.1411499111
[42]	Tan, H. Y.; Wang, N.; Li, S.; Hong, M.; Wang, X.; Feng, Y. Oxid. Med. Cell. Longev. 2016, 279, 5090.
[43]	Chen, L.; Deng, H.; Cui, H.; Fang, J.; Zuo, Z.; Deng, J.; Li, Y.; Wang, X.; Zhao, L. Oncotarget. 2018, 9, 7204. doi: 10.18632/oncotarget.23208
[44]	Matsumoto, A.; Sato, N.; Kataoka, K.; Miyahara, Y. J. Am. Chem. Soc. 2009, 131, 12022. doi: 10.1021/ja902964m pmid: 19663494
[45]	Wang, J.; Wu, W.; Zhang, Y.; Wang, X.; Qian, H.; Liu, B.; Jiang, X. Biomaterials. 2014, 35, 866. doi: 10.1016/j.biomaterials.2013.10.028
[46]	Wang, H.; Wang, K.; Sun, J.; Fang, G. Q.; Yao, Q. Q.; Wu, Z. Y. Chinese J. Org. Chem. 2018, 38, 1035. (in Chinese) doi: 10.6023/cjoc201709037
	(王浩, 王凯, 孙捷, 方桂迁, 姚庆强, 吴忠玉, 有机化学, 2018, 38, 1035.) doi: 10.6023/cjoc201709037
[47]	Cao, Z. B.; Chen, X.; Yao, J. R.; Huang, L.; Shao, Z. Z. Soft Matter. 2007, 3, 910. doi: 10.1039/b703139d

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