Liposomal drug delivery has become an established technology platform to deliver dual drugs to produce synergistic effects and reduce the adverse effects of traditional chemotherapy. Gambogic acid (GA) and retinoic acid (RA) are both effective anticancer components, but their low water-solubility (gambogic acid < 0.0050 mg/mL, retinoic acid 0.0048 < mg/mL) makes it difficult to load both drugs into the liposomes actively using the conventional method. We have successfully used solvent-assisted active loading technology (SALT) to load the insoluble drugs into the internal water phase via water-miscible organic solvent. Gambogic acid and retinoic acid co-encapsulated liposomes (weight ratio of GA to RA = 1:2, GRL) exhibited the strongest synergistic effect; combination index (CI) was 0.614 in 4T1 cells. Our studies demonstrated that GRL had uniform droplet size of about 130 nm, high stability, and controlled release behavior. GRL outperformed gambogic acid and retinoic acid solution (GRS) in pharmacokinetic profiles for a longer half-life and increased AUC. Comparing to GRS, GL, and RL, GRL showed increased cytotoxicity and apoptosis in 4T1 cells and showed the strongest anti-tumor ability in the in vivo anti-tumor efficacy. Overall, the SALT was a promising method to active loading poorly soluble drugs into liposomes, and the results showed GRL possessed a great potential for use in synergistic anticancer therapy.
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This work was financially supported by the Career Development Program for Yong and Middle-aged Teachers in Shenyang Pharmaceutical University.
Compliance with ethical standards
All procedures performed in studies involving animals were in accordance with the national regulations and were approved by Institutional Animal Ethical Care Committee of Shenyang Pharmaceutical University (SYPU-IACUC-C2018-4-2-203, approval date: 2 April 2018).
Conflict of interest
The authors declare that they have no conflict of interest.
Doddapaneni R, Patel K, Owaid IH, Singh M. Tumor neovasculature-targeted cationic PEGylated liposomes of gambogic acid for the treatment of triple-negative breast cancer. Drug Deliv. 2016;23(4):1–10.CrossRefGoogle Scholar
Jiang LL, Kang L, Lin QH, Jian R, He ZH, Li H, et al. Gambogic acid causes fin developmental defect in zebrafish embryo partially via retinoic acid signaling. Reprod Toxicol. 2016;63:161–8.CrossRefPubMedGoogle Scholar
Cristiano MC, Cosco D, Celia C, Tudose A, Mare R, Paolino D, et al. Anticancer activity of all-trans retinoic acid-loaded liposomes on human thyroid carcinoma cells. Colloids Surf B: Biointerfaces. 2016;150:408.CrossRefPubMedGoogle Scholar
Kawakami S, Suzuki S, Yamashita F, Hashida M. Induction of apoptosis in A549 human lung cancer cells by all- trans retinoic acid incorporated in DOTAP/cholesterol liposomes. J Control Release. 2006;110(3):514–21.CrossRefPubMedGoogle Scholar
Yao J, Li Y, Sun X, Dahmani FZ, Liu H, Zhou J. Nanoparticle delivery and combination therapy of gambogic acid and all-trans retinoic acid. Int J Nanomedicine. 2014;2014(Issue 1):3313–24.CrossRefGoogle Scholar
Liu L, Qi XJ, Zhong ZK, Zhang EN. Nanomedicine-based combination of gambogic acid and retinoic acid chlorochalcone for enhanced anticancer efficacy in osteosarcoma. Biomed Pharmacother. 2016;83:79–84.CrossRefPubMedGoogle Scholar
Cheng Y, Zhao P, Wu S, Yang T, Chen Y, Zhang X, et al. Cisplatin and curcumin co-loaded nano-liposomes for the treatment of hepatocellular carcinoma. Int J Pharm. 2018;545(1–2):261–73.CrossRefPubMedGoogle Scholar
He RX, Ye X, Li R, Chen W, Ge T, Huang TQ, et al. PEGylated niosomes-mediated drug delivery systems for paeonol:preparation, pharmacokinetics studies and synergistic anti-tumor effects with 5-FU. J Liposome Res. 2016;27(2):161–70.CrossRefPubMedGoogle Scholar
Jiang K, Shen M, Xu W. Arginine, glycine, aspartic acid peptide-modified paclitaxel and curcumin co-loaded liposome for the treatment of lung cancer: in vitro/vivo evaluation. Int J Nanomedicine. 2018;13:2561–9.CrossRefPubMedPubMedCentralGoogle Scholar
Yu Z, Chen Q, Yang Y, Lin X, Ma W, Chen G et al. Platinum (IV) prodrugs with long lipid chains for drug delivery and overcoming cisplatin resistance. Chem Commun 2018;54(42):https://doi.org/10.1039/C8CC02791A.
Langton MJ, Scriven LM, Williams NH, Hunter CA. Triggered release from lipid bilayer vesicles by an artificial transmembrane signal transduction system. J Am Chem Soc. 2017;139(44):15768–73.CrossRefPubMedGoogle Scholar
Hu CMJ, Zhang L. Nanoparticle-based combination therapy toward overcoming drug resistance in cancer. Biochem Pharmacol. 2012;83(8):1104–11.CrossRefPubMedGoogle Scholar
Liu Y, Fang J, Kim YJ, Wong MK, Wang P. Codelivery of doxorubicin and paclitaxel by cross-linked multilamellar liposome enables synergistic antitumor activity. Mol Pharm. 2014;11(5):1651–61.CrossRefPubMedPubMedCentralGoogle Scholar
Han X, Chen J, Jiang M, Zhang N, Na K, Luo C, et al. Paclitaxel-paclitaxel prodrug nanoassembly as a versatile nanoplatform for combinational cancer therapy. ACS Appl Mater Interfaces. 2016;8(49):33506–13.CrossRefPubMedGoogle Scholar
Yang Y, Lu X, Liu Q, Dai Y, Zhu X, Wen Y, et al. Palmitoyl ascorbate and doxorubicin co-encapsulated liposome for synergistic anticancer therapy. Eur J Pharm Sci. 2017;105:219–29.CrossRefPubMedGoogle Scholar
Liu Y, Tamam H, Yeo Y. Mixed liposome approach for ratiometric and sequential delivery of paclitaxel and gemcitabine. AAPS PharmSciTech. 2017;141(2):1–7.Google Scholar
Lu L, Du Y, Ismail M, Ling L, Yao C, Fu Z et al. Liposomes assembled from dimeric retinoic acid phospholipid with improved pharmacokinetic properties. Eur J Pharm Sci 2017.Google Scholar
Tefas LR, Sylvester B, Tomuta I, Sesarman A, Licarete E, Banciu M, et al. Development of antiproliferative long-circulating liposomes co-encapsulating doxorubicin and curcumin, through the use of a quality-by-design approach. Drug Des Devel Ther. 2017;11:1605–21.CrossRefPubMedPubMedCentralGoogle Scholar
Gubernator J. Active methods of drug loading into liposomes: recent strategies for stable drug entrapment and increased in vivo activity. Expert Opin Drug Deliv. 2011;8(5):565–80.CrossRefPubMedGoogle Scholar
Eloy JO, Petrilli R, Chesca DL, Saggioro FP, Lee RJ, Marchetti JM. Anti-HER2 immunoliposomes for co-delivery of paclitaxel and rapamycin for breast cancer therapy. Eur J Pharm Biopharm. 2017;115:159–67.CrossRefPubMedGoogle Scholar
Modi S, Xiang TX, Anderson BD. Enhanced active liposomal loading of a poorly soluble ionizable drug using supersaturated drug solutions. J Control Release. 2012;162(2):330–9.CrossRefPubMedGoogle Scholar
Hayes ME, Noble CO, Szoka FC, inventors; Remote loading of sparingly water-soluble drugs into liposomes 2015.Google Scholar
Tang WL, Tang WH, Szeitz A, Kulkarni J, Cullis P, Li SD. Systemic study of solvent-assisted active loading of gambogic acid into liposomes and its formulation optimization for improved delivery. Biomaterials. 2018;166:13–26.CrossRefPubMedGoogle Scholar
Tang WL, Chen WC, Roy A, Undzys E, Li SD. A simple and improved active loading method to efficiently encapsulate Staurosporine into lipid-based nanoparticles for enhanced therapy of multidrug resistant cancer. Pharm Res. 2016;33(5):1104–14.CrossRefPubMedGoogle Scholar
Avnir Y, Ulmansky R, Wasserman V, Evenchen S, Broyer M, Barenholz Y, et al. Amphipathic weak acid glucocorticoid prodrugs remote-loaded into sterically stabilized nanoliposomes evaluated in arthritic rats and in a Beagle dog: a novel approach to treating autoimmune arthritis. Arthritis Rheum. 2008;58(1):119–29.CrossRefPubMedGoogle Scholar
Clerc S, Barenholz Y. Loading of amphipathic weak acids into liposomes in response to transmembrane calcium acetate gradients. Biochim Biophys Acta. 1995;1240(2):257–65.CrossRefPubMedGoogle Scholar
Adamson PC, Balis FM, Smith MA, Murphy RF, Godwin KA, Poplack DG. Dose-dependent pharmacokinetics of all-trans-retinoic acid. J Natl Cancer Inst. 1992;84(17):1332–5.CrossRefPubMedGoogle Scholar
Hao K, Zhao XP, Liu XQ, Wang GJ. Determination of gambogic acid in dog plasma by high-performance liquid chromatography for a pharmacokinetic study. Biomed Chromatogr. 2007;21(3):279–83.CrossRefPubMedGoogle Scholar
Hou L, Yao J, Zhou J. Simultaneous LC–MS analysis of paclitaxel and retinoic acid in plasma and tissues from tumor-bearing mice. Chromatographia. 2011;73(5–6):471–80.CrossRefGoogle Scholar