Nanoparticle delivery of chemotherapy combination regimen improves the therapeutic efficacy in mouse models of lung cancer Jing Tian, PhD, Yuangzeng.

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Nanoparticle delivery of chemotherapy combination regimen improves the therapeutic efficacy in mouse models of lung cancer Jing Tian, PhD, Yuangzeng Min, PhD, Zachary Rodgers, PhD, Xiaomeng Wan, B.S, Hui Qiu, M.S, Yu Mi, PhD, Xi Tian, PhD, Kyle T. Wagner, B.S, Joseph M. Caster, MD, PhD, Yanfei Qi, PhD, Kyle Roche, PhD, Tian Zhang, MD, Jianjun Cheng, PhD, Andrew Z. Wang, MD Nanomedicine: Nanotechnology, Biology and Medicine Volume 13, Issue 3, Pages 1301-1307 (April 2017) DOI: 10.1016/j.nano.2016.11.007 Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 1 CPP and DTX co-delivery from PLGA–PEG NPs. NP co-encapsulation allows for accurate exposure of the murine tumor site to both drugs whereas freely dosed drugs may lead to variations in tumor-drug exposure and reductions in potency. Nanomedicine: Nanotechnology, Biology and Medicine 2017 13, 1301-1307DOI: (10.1016/j.nano.2016.11.007) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 2 (A) Drug loading (wt%) and (B) encapsulation efficiency (%EE) of the singly drug-loaded C4CP, C8CP, and C10CP NPs. PLGA–PEG NPs were formed by nanoprecipitation in the presence of CPPs at different %FR. After NP formation and washing, final drug loadings were determined using digestion followed by HPLC analysis. n.s. indicates no significant difference; * indicates P<0.05; ** indicates P<0.01; *** indicates P<0.001. Nanomedicine: Nanotechnology, Biology and Medicine 2017 13, 1301-1307DOI: (10.1016/j.nano.2016.11.007) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 3 Drug loading (wt%) and encapsulation efficiency (% EE) of dually loaded NPs. PLGA–PEG NPs were formed with a constant 10%FR DTX and various %FR of (A, B) C4CP, (C, D) C8CP, or (E, F) C10CP. The ratios shown above each bar in panels A, C, and E correspond to the DTX:CPP molar ratio encapsulated in the NPs. Nanomedicine: Nanotechnology, Biology and Medicine 2017 13, 1301-1307DOI: (10.1016/j.nano.2016.11.007) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 4 Release kinetics of CPPs from (A) singly or (B) dually loaded PLGA–PEG NPs, and (C) DTX release from DTX containing NPs under physiological sink conditions. Loaded NPs were dialyzed against a large excess of PBS and the NPs' retained drug was determined using HPLC after digestion with acetonitrile. ** indicates P<0.01; *** indicates P<0.001. Nanomedicine: Nanotechnology, Biology and Medicine 2017 13, 1301-1307DOI: (10.1016/j.nano.2016.11.007) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 5 In vitro cytotoxicity IC50 values of free and encapsulated small molecule chemotherapeutics in a non-small cell (H460) and small cell lung cancer (344SQ) line. Cells were treated with of either free drugs, free drugs in combination, singly loaded NPs, singly loaded NPs in combination (DTX NPs+CPP NPs), or dually loaded NPs (DTX:CPP NPs). * indicates P<0.05; ** indicates P<0.01; *** indicates P<0.001. Nanomedicine: Nanotechnology, Biology and Medicine 2017 13, 1301-1307DOI: (10.1016/j.nano.2016.11.007) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 6 Free drug and NP formulation in vivo efficacy represented by tumor volume change in (A) 344SQ or (B) H460 murine xenograft models. Mice were treated with combinations of either the free drugs, singly loaded NPs (DTX NP+CPP NP), or dually loaded NPs (DTX:C8CP). n.s. indicates no significant difference; * indicates P<0.05, a indicates P=0.051. Nanomedicine: Nanotechnology, Biology and Medicine 2017 13, 1301-1307DOI: (10.1016/j.nano.2016.11.007) Copyright © 2016 Elsevier Inc. Terms and Conditions

Nanomedicine: Nanotechnology, Biology and Medicine 2017 13, 1301-1307DOI: (10.1016/j.nano.2016.11.007) Copyright © 2016 Elsevier Inc. Terms and Conditions