Liposomes decorated with tumour-targeting cell-penetrating peptides can enhance specific drug delivery at the tumour site. The TR peptide, c(RGDfK)-AGYLLGHINLHHLAHL(Aib)HHIL, is pH-sensitive and actively targets tumour cells that overexpress integrin receptor αvβ3, such as B16F10 melanoma cells. Liposomes can be modified with the TR peptide by two different methods: utilization of the cysteine residue on TR to link DSPE-PEG2000-Mal contained in the liposome formula (LIPTR) or decoration of TR with a C18 stearyl chain (C18-TR) for direct insertion into the liposomal phospholipid bilayer through electrostatic and hydrophobic interactions (LIPC18-TR). We found that both TR and C18-TR effectively reversed the surface charge of the liposomes when the systems encountered the low pH of the tumour microenvironment, but LIPC18-TR exhibited a greater increase in the charge, which led to higher cellular uptake efficiency. Correspondingly, the IC50 values of PTX-LIPTR and PTX-LIPC18-TR in B16F10 cells in vitro were 2.1-fold and 2.5-fold lower than that of the unmodified PTX-loaded liposomes (PTX-LIP), respectively, in an acidic microenvironment (pH 6.3). In B16F10 tumour-bearing mice, intravenous administration of PTX-LIPTR and PTX-LIPC18-TR (8 mg/kg PTX every other day for a total of 4 injections) caused tumour reduction ratios of 39.4% and 56.1%, respectively, compared to 20.8% after PTX-LIP administration. Thus, we demonstrated that TR peptide modification could improve the antitumour efficiency of liposomal delivery systems, with C18-TR presenting significantly better results. After investigating different modification methods, our data show that selecting an adequate method is vital even when the same molecule is used for decoration.
This is a preview of subscription content, access via your institution
Subscribe to Journal
Get full journal access for 1 year
only $9.92 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
Vaupel P, Kallinowski F, Okunieff P. Blood flow, oxygen and nutrient supply, and metabolic microenvironment of human tumors: a review. Cancer Res. 1989;49:6449–65.
Tannock IF, Rotin D. Acid pH in tumors and its potential for therapeutic exploitation. Cancer Res. 1989;49:4373–84.
Wojtkowiak JW, Verduzco D, Schramm KJ, Gillies RJ. Drug resistance and cellular adaptation to tumor acidic pH microenvironment. Mol Pharm. 2011;8:2032–8.
Tian L, Bae YH. Cancer nanomedicines targeting tumor extracellular pH. Colloids Surf B Biointerfaces. 2012;99:116–26.
Béduneau A, Saulnier P, Benoit JP. Active targeting of brain tumors using nanocarriers. Biomaterials. 2007;28:4947–67.
Sudimack J, Lee RJ. Targeted drug delivery via the folate receptor. Adv Drug Deliv Rev. 2000;41:147–62.
Vugrin D, Whitmore WF Jr, Sogani PC, Bains M, Herr HW, Golbey RB. Combined chemotherapy and surgery in treatment of advanced germ-cell tumors. Cancer. 1981;47:2228–31.
Tormey DC. Combined chemotherapy and surgery in breast cancer: a review. Cancer. 1975;36:881–92.
Morise Z, Sugioka A, Tokoro T, Tanahashi Y, Okabe Y, Kagawa T, et al. Surgery and chemotherapy for intrahepatic cholangiocarcinoma. World J Hepatol. 2010;2:58–64.
Markman M, Mekhail TM. Paclitaxel in cancer therapy. Expert Opin Pharmacother. 2002;3:755–66.
Rowinsky EK, Donehower RC. Paclitaxel (taxol). N. Engl J Med. 1995;332:1004–14.
Spencer CM, Faulds D. Paclitaxel. Drugs. 1994;48:794–847.
Wang LY, Zhou BJ, Huang SQ, Qu MK, Lin Q, Gong T, et al. Novel fibronectin-targeted nanodisk drug delivery system displayed superior efficacy against prostate cancer compared with nanospheres. Nano Res. 2019;12:2451–9.
Yang T, Cui FD, Choi MK, Cho JW, Chung SJ, Shim CK, et al. Enhanced solubility and stability of PEGylated liposomal paclitaxel: in vitro and in vivo evaluation. Int J Pharm. 2007;338:317–26.
Koudelka Š, Turánek J. Liposomal paclitaxel formulations. J Control Rel. 2012;163:322–34.
Koren E, Torchilin VP. Cell-penetrating peptides: breaking through to the other side. Trends Mol Med. 2012;18:385–93.
Juliano RL, Alam R, Dixit V, Kang HM. Cell-targeting and cell-penetrating peptides for delivery of therapeutic and imaging agents. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2009;1:324–35.
Copolovici DM, Langel K, Eriste E, Langel Ü. Cell-penetrating peptides: design, synthesis, and applications. ACS Nano. 2014;8:1972–94.
Torchilin VP. Cell-penetrating peptide-modified pharmaceutical nanocarriers for intracellular drug and gene delivery. Biopolymers. 2008;90:604–10.
Snyder EL, Dowdy SF. Cell-penetrating peptides in drug delivery. Pharm Res. 2004;21:389–93.
Torchilin VP, Lukyanov AN. Peptide and protein drug delivery to and into tumors: challenges and solutions. Drug Discov Today. 2003;8:259–66.
Fonseca SB, Pereira MP, Kelley SO. Recent advances in the use of cell-penetrating peptides for medical and biological applications. Adv Drug Deliv Rev. 2009;61:953–64.
Mondal G, Barui S, Chaudhuri A. The relationship between the cyclic-RGDfK ligand and αvβ3 integrin receptor. Biomaterials. 2013;34:6249–60.
Sakurai Y, Hatakeyama H, Sato Y, Hyodo M, Akita H, Ohga N, et al. RNAi-mediated gene knockdown and anti-angiogenic therapy of RCCs using a cyclic RGD-modified liposomal-siRNA system. J Control Rel. 2014;173:110–8.
Shi KR, Li JP, Cao ZL, Yang P, Qiu Y, Yang B, et al. A pH-responsive cell-penetrating peptide-modified liposomes with active recognizing of integrin αvβ3 for the treatment of melanoma. J Control Rel. 2015;217:138–50.
Song WT, Tang ZH, Zhang DW, Zhang Y, Yu HY, Li MQ, et al. Anti-tumor efficacy of c (RGDfK)-decorated polypeptide-based micelles co-loaded with docetaxel and cisplatin. Biomaterials. 2014;35:3005–14.
Wang LY, Qu MK, Huang SQ, Fu Y, Yang LQ, He SS, et al. A novel α-enolase-targeted drug delivery system for high efficacy prostate cancer therapy. Nanoscale. 2018;10:13673–83.
Jiang TY, Mo R, Bellotti A, Zhou JP, Gu Z. Gel–liposome-mediated co-delivery of anticancer membrane-associated proteins and small-molecule drugs for enhanced therapeutic efficacy. Adv Funct Mater. 2014;24:2295–304.
Jiang TY, Wang T, Li T, Ma YD, Shen SY, He BF, et al. Enhanced transdermal drug delivery by transfersome-embedded oligopeptide hydrogel for topical chemotherapy of melanoma. ACS Nano. 2018;12:9693–701.
Song X, Wan ZY, Chen TJ, Fu Y, Jiang KJ, Yi XL, et al. Development of a multi-target peptide for potentiating chemotherapy by modulating tumor microenvironment. Biomaterials. 2016;108:44–56.
Zhang XM, Zhang Q, Peng Q, Zhou J, Liao LF, Sun X, et al. Hepatitis B virus preS1-derived lipopeptide functionalized liposomes for targeting of hepatic cells. Biomaterials. 2014;35:6130–41.
Zhang QY, Tang J, Fu L, Ran R, Liu YY, Yuan MQ, et al. A pH-responsive α-helical cell-penetrating peptide-mediated liposomal delivery system. Biomaterials. 2013;34:7980–93.
Yang T, Choi MK, Cui FD, Kim JS, Chung SJ, Shim CK, et al. Preparation and evaluation of paclitaxel-loaded PEGylated immunoliposome. J Control Rel. 2007;120:169–77.
Qu MK, Lin Q, He SS, Wang LY, Fu Y, Zhang ZR, et al. A brain targeting functionalized liposomes of the dopamine derivative N-3, 4-bis (pivaloyloxy)-dopamine for treatment of Parkinson’s disease. J Control Rel. 2018;277:173–82.
Huang SQ, Deng L, Zhang HM, Wang LY, Zhang YC, Lin Q, et al. Co-delivery of TRAIL and paclitaxel by fibronectin-targeting liposomal nanodisk for effective lung melanoma metastasis treatment. Nano Res. 2022;15:728–37.
Khalil IA, Kogure K, Futaki S, Hama S, Akita H, Ueno M, et al. Octaarginine-modified multifunctional envelope-type nanoparticles for gene delivery. Gene Ther. 2007;14:682.
Jang B, Park JY, Tung CH, Kim IH, Choi Y. Gold nanorod-photosensitizer complex for near-infrared fluorescence imaging and photodynamic/photothermal therapy in vivo. ACS Nano. 2011;5:1086–94.
Lammers T, Kiessling F, Hennink WE, Storm G. Drug targeting to tumors: principles, pitfalls and (pre-) clinical progress. J Control Rel. 2012;161:175–87.
Bigini P, Previdi S, Casarin E, Silvestri D, Violatto MB, Facchin S, et al. In vivo fate of avidin-nucleic acid nanoassemblies as multifunctional diagnostic tools. ACS Nano. 2014;8:175–87.
Gao YJ, Zhou YX, Zhao L, Zhang C, Li YS, Li JW, et al. Enhanced antitumor efficacy by cyclic RGDyK-conjugated and paclitaxel-loaded pH-responsive polymeric micelles. Acta Biomater. 2015;23:127–35.
Al-Jamal KT, Al-Jamal WT, Wang JT, Rubio N, Buddle J, Gathercole D, et al. Cationic poly-L-lysine dendrimer complexes doxorubicin and delays tumor growth in vitro and in vivo. ACS Nano. 2013;7:1905–17.
Xu ZH, Wang YH, Zhang L, Huang L. Nanoparticle-delivered transforming growth factor-beta siRNA enhances vaccination against advanced melanoma by modifying tumor microenvironment. ACS Nano. 2014;8:3636–45.
Huang SQ, Zhang YC, Wang LY, Liu W, Xiao LY, Lin Q, et al. Improved melanoma suppression with target-delivered TRAIL and Paclitaxel by a multifunctional nanocarrier. J Control Rel. 2020;325:10–24.
Salvati A, Pitek AS, Monopoli MP, Prapainop K, Bombelli FB, Hristov DR, et al. Transferrin-functionalized nanoparticles lose their targeting capabilities when a biomolecule corona adsorbs on the surface. Nat Nanotechnol. 2013;8:137–43.
Spinks CB, Zidan AS, Khan MA, Habib MJ, Faustino PJ. Pharmaceutical characterization of novel tenofovir liposomal formulations for enhanced oral drug delivery: in vitro pharmaceutics and Caco-2 permeability investigations. Clin Pharmacol. 2017;9:29.
Lainé AL, Gravier J, Henry M, Sancey L, Béjaud J, Pancani E, et al. Conventional versus stealth lipid nanoparticles: Formulation and in vivo fate prediction through FRET monitoring. J Control Rel. 2014;188:1–8.
Xiao D, Jia HZ, Ma N, Zhuo RX, Zhang XZ. A redox-responsive mesoporous silica nanoparticle capped with amphiphilic peptides by self-assembly for cancer-targeting drug delivery. Nanoscale. 2015;7:10071–7.
Yu YC, Tirrell M, Fields GB. Minimal lipidation stabilizes protein-like molecular architecture. J Am Chem Soc. 1998;120:9979–87.
Forns P, Lauer‐Fields JL, Gao S, Fields GB. Induction of protein-like molecular architecture by monoalkyl hydrocarbon chains. Biopolymers. 2000;54:531–46.
Han J, Huang X, Sun LD, Li Z, Qian H, Huang WL. Novel fatty chain-modified glucagon-like peptide-1 conjugates with enhanced stability and prolonged in vivo activity. Biochem Pharmacol. 2013;86:297–308.
Zhang HY, Schneider SE, Bray BL, Friedrich PE, Tvermoes NA, Mader CJ, et al. Process development of TRI-999, a fatty-acid-modified HIV fusion inhibitory peptide. Org Process Res Dev. 2008;12:101–10.
This work was supported by the National Natural Science Foundation of China (Nos. 81690261, 82022070 and 81872824) and the Medico-Engineering Cooperation Programme from the Med-X Center for Materials, Sichuan University (MCM202103).
The authors declare no competing interests.
About this article
Cite this article
Huang, Sq., Zhang, Hm., Zhang, Yc. et al. Comparison of two methods for tumour-targeting peptide modification of liposomes. Acta Pharmacol Sin (2022). https://doi.org/10.1038/s41401-022-01011-4
- cell-penetrating peptides
- B16F10 melanoma cells