The use of biotinylated-EGF-modified gelatin nanoparticle carrier to enhance cisplatin accumulation in cancerous lungs via inhalation Ching-Li Tseng a , Wen-Yun Su a, b , Ko-Chung Yen b , Kai-Chiang Yang b , Feng-Huei Lin a, b, * a Division of Medical Engineering Research, National Health Research Institutes, No.35, Keyan Rd., Zhunan Town, Miaoli County 350, Taiwan, ROC b Institute of Biomedical Engineering, National Taiwan University, No.1, Sec.1, Ren-ai Rd., Taipei City 100, Taiwan, ROC article info Article history: Received 4 February 2009 Accepted 7 March 2009 Available online 5 April 2009 Keywords: Gelatin nanoparticle (GPs) Cisplatin (CDDP) Lung cancer Anticancer activity Epidermal growth factor (EGF) Inhalation abstract To develop a polymer-anticancer drug conjugate, we employed gelatin nanoparticles (GPs) as carriers of cisplatin (CDDP) with anticipated improved therapeutic effect and reduced side effects. The anticancer activities of CDDP-incorporated in GPs (GP–Pt) with biotinylated-EGF (bEGF) modification (GP–Pt–bEGF) were studied. GP–Pt–bEGF with EGFR affinity produced much higher Pt concentrations in A549 cells (high EGFR expression) than in HFL1 cells (low EGFR expression). An in vitro anticancer study showed that GP–Pt–bEGF was more potent than free CDDP or GP–Pt because of its rapid effect on the cell cycle as well as a lower IC 50 (1.2 mg/ml) that inhibits A549 cell growth. PI staining showed that cells treated with GP–Pt-bEGF for only 4 h had the highest sub-G1 population. The CDDP formulations – free CDDP, GP–Pt, and GP–Pt–bEGF – were given by intratumorous injections to SCID mice in a subcutaneous model. This treatment showed that GP–Pt–bEGF had stronger anti-tumor activity and was less toxic than free CDDP in vivo. Mice treated with GP–Pt–bEGF showed slight body weight loss, whereas free CDDP treatment at the same dose caused a body weight loss of 20–30%. Furthermore, these formulations were given to mice with lung cancer via aerosol delivery. This treatment showed that inhaled GP–Pt–bEGF could target EGFR-overexpressing cells to achieve high cisplatin dosage in cancerous lungs. To summarize, gelatin nanoparticles loaded with CDDP and decorated with EGF tumor-specific ligand were successfully developed. Their in vitro and in vivo targeting ability and anticancer effect were confirmed. The aerosol delivery of the nanodrug carrier was demonstrated. Simple aerosol delivery of targeted drug carriers may prove useful for the clinical treatment of lung cancer patients. Ó 2009 Elsevier Ltd. All rights reserved. 1. Introduction Lung cancer is one of the most harmful forms of cancer. The long-term survival rate of lung cancer patients treated by conven- tional modalities, such as surgical resection, radiation, and chemotherapy remains far from satisfactory. Systemic drug delivery is rarely successful because only a limited dosage of the chemotherapeutic drugs target lung tumor sites, even when administered at a high dose [1]. Most chemotherapeutic drugs act on normal cells, inhibiting their growth; this makes the patient extremely weak and can even result in death. Cisplatin (cis-dichlorodiammineplatinum (II); CDDP) is a key drug used in the chemotherapy of cancers, including gastrointestinal, genitourinary and lung cancer; cisplatinum-based chemotherapy is accepted as a standard first-line treatment for advanced non-small cell lung carcinoma (NSCLC) [2]. Chemo- therapy with cisplatin is associated with various secondary effects, such as anemia, nausea, vomiting, neurotoxicity and nephrotoxicity [3], which impair the patient’s quality of life and can even be life- threatening in preexisting conditions. Due to these side effects, alternative methods of administering toxic cisplatin are needed. For the reduction of side effects, specific drug delivery systems (DDSs) have been investigated. Several polymers, of natural as well as synthetic origin, have been used as reservoirs for delivery of cisplatin, such as polymeric micelles [3], poly(g,L-glutamic acid)- (g-PGA) [4], polylactic acid (PLA) [5], etc. In this study, gelatin was selected as the raw material to prepare the drug carrier because of its biocompatibility and biodegradability [6,7]. Its degradation rate can be regulated by the degree of crosslinking to control drug release rate. Konishi et al. showed that a gelatin hydrogel con- taining CDDP could release the drug over a long period, resulting in a superior anticancer effect [8]. However, there have been few * Corresponding author at. Division of Medical Engineering Research, National Health Research Institutes, No.35, Keyan Rd, Zhunan Town, Miaoli county 350, Taiwan, ROC. Tel.: þ886 37 246166x37100; fax: þ886 37 586440. E-mail address: double@nhri.org.tw (F.-H. Lin). Contents lists available at ScienceDirect Biomaterials journal homepage: www.elsevier.com/locate/biomaterials 0142-9612/$ – see front matter Ó 2009 Elsevier Ltd. All rights reserved. doi:10.1016/j.biomaterials.2009.03.010 Biomaterials 30 (2009) 3476–3485