米托蒽醌隐形阳离子脂质体的制备及体内外抗肿瘤活性

目的:制备米托蒽醌隐形阳离子脂质体(YM),并研究其制剂学性质及体内外抗肿瘤作用。方法:膜材料中引入2-二油酰基羟丙基-3-N,N,N-三甲铵氯(DOTAP)和聚乙二醇2000-二硬脂酰磷脂乙醇胺(DSPE-PEG₂₀₀₀)分别作为阳性和隐形膜材,采用硫酸铵梯度法制备YM,并以泄漏率为指标,考察其稳定性;高内涵扫描仪考察2 h时其在B16F10细胞中的摄取情况;MTT 法测定其对B16F10细胞的增殖影响;B16F10皮下移植瘤模型研究其体内抗肿瘤作用。结果:YM的粒径和Zeta电位分别为(118.2±4.6)nm和(31.6±4.9)mV;载药量和包封率分别为(6.5±0.2)%和(96.2±1.8)%;血清中48 h累积泄漏率<30%,稳定性良好;YM的细胞摄取量是普通长循环脂质体(CM)的10倍,IC₅₀与CM相比显著降低(P<0.05),体内抑瘤率为58.8%,显著高于CM组的42.7%。结论:YM稳定性良好,具有较好的体内外抗肿瘤活性。     

DOPA-based paclitaxel-loaded liposomes with modifications of transferrin and alendronate for bone and myeloma targeting

Treatment for multiple myeloma (MM) with a combined strategy of bone and tumor targeting remains a crucial technical challenge due to the incorporation of various functional components into one single system. Here, we developed dioleoyl phosphatidic acid (DOPA)-based paclitaxel (PTX)-loaded liposomes with modifications of alendronate and transferrin (Ald-/Tf-modified PTX-L), which were capable of bone affinity mediated by phosphate groups in DOPA and alendronate, and tumor targeting offered by transferrin. Ald-/Tf-modified PTX-L had clear and well-defined spherical shape with an intermediated size of 118.8?±?4.8?nm, a highly negative surface charge of ?46.9?±?6.8?mV and a drug entrapment efficiency (DEE) of approximately 80%. When the pH was changed from pH 7.4 to pH 6.5, the accumulative release of PTX from Ald-/Tf-modified PTX-L significantly increased from 26.7?±?3.7% to 41.7?±?4.9%. Importantly, liposomes based on DOPA displayed an obviously stronger affinity with hydroxyapatite (HAp) than 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE)-based liposomes. Compared to PTX-L, Ald-/Tf-modified PTX-L exhibited obvious improvement of cytotoxicity (IC_50?=_?1.25?±?0.09?μg/mL), significant enhancement on PTX intracellular accumulation (16.58?±?0.62?μg/mg) and notable promotion to apoptosis induction (45.21?±?3.10%) toward myeloma (MM1s) cells. In this study of antitumor efficacy, Ald-/Tf-modified PTX-L with bone-specific targeting showed a significant effect on extending the median survival time (48 days) and terminal survival time (>?58 days) against the MM1S-injected nude mice among all formulations. The results suggested that Ald-/Tf-modified PTX-L had potential as an efficient anticancer drug delivery system for MM therapy.     

Hydroxycamptothecin liposomes based on thermal and magnetic dual-responsive system: preparation,in vitro and in vivo antitumor activity,microdialysis-based tumor pharmacokinetics

Due to the absence of lactone form of hydroxycamptothecin, the commercially available hydroxycamptothecin injection exhibits inefficient therapeutic effects. In this study, we constructed a novel delivery system (thermosensitive magnetic liposomes) that protects lactone form of hydroxycamptothecin from blood or water. After hydroxycamptothecin was loaded into the thermosensitive magnetic liposome (HCPT/TML), its in vitro and in vivo antitumor activity and microdialysis-based tumour pharmacokinetics were determined. The results demonstrated that HCPT/TMLs possessed favourable physicochemical features and significant cytotoxicity against the Huh-7 cells in vitro. In the in vivo antitumor study and tumour pharmacokinetics, HCPT/TMLs displayed effective targeting delivery and antitumor effects, which corresponded to the determined hydroxycamptothecin concentration in tumour tissue. In conclusion, this thermal and magnetic dual-responsive system can efficiently deliver hydroxycamptothecin to tumour tissue and has great potential application in cancer treatment.     

提高人肺腺癌裸鼠皮下移植瘤成瘤率的方法学研究

目的：提高人肺腺癌裸鼠皮下移植瘤的成瘤率。方法：人肺腺癌SPC-A-1、GLC-82培养细胞或相应的瘤组织匀浆接种于裸鼠皮下之前,先用富含生长因子的小鼠肉瘤S180腹水上清液按比例稀释,然后在裸鼠背部或腋窝皮下接种。观察人肿瘤细胞或肿瘤组织在加入小鼠肉瘤S180腹水上清液后,对移植瘤生长速度及成瘤率的影响。结果：加入小鼠肉瘤S180腹水上清液后,人肺腺癌SPC-A-1和GLC-82成瘤率明显提高,而且移植后的人肿瘤生长速度较未加小鼠肉瘤S180腹水上清液的人肺腺癌细胞明显加快。结论：小鼠肉瘤S180腹水上清液能明显提高人肺腺癌SPC-A-1和GLC-82裸鼠皮下移植瘤的成瘤率,促进肿瘤生长。     

Antitumor activity of RT2 peptide derived from crocodile leukocyte peptide on human colon cancer xenografts in nude mice

RT2, derived from the leukocyte peptide of Crocodylus siamensis, can kill human cervical cancer cells via apoptosis induction, but no evidence has shown in vivo. In this study, we investigated the antitumor effect of RT2 on human colon cancer xenografts in nude mice. Twenty‐four mice were injected subcutaneously with human colon cancer HCT 116 cells. Eleven days after cancer cell implantation, the mice were treated with intratumoral injections of phosphate buffered saline (PBS) or RT2 (0.01, 0.1, and 1 mg/mouse) once every 2 days for a total of 5 times. The effect of a 10‐day intratumoral injection of RT2 on body weight, biochemical, and hematological parameters in BALB/c mice showed no significant difference between the groups. Tumor volume showed a significant decrease only in the treatment group with RT2 (1 mg/mouse) at day 6 (P < .05), day 8 (P < .01), and day 10 (P < .01) after the first treatment. The protein expression levels of cleaved poly (ADP‐ribose) polymerase (PARP), apoptosis‐inducing factor (AIF), and the p53 tumor suppressor protein (p53) in xenograft tumors increased after treatment with RT2 (1 mg/mouse) compared to those in the PBS‐injected group. Moreover, RT2 increased the expression of Endo G and Bcl‐2 family proteins. Therefore, the peptide RT2 can inhibit tumor growth via the induction of apoptosis in an in vivo xenograft model.     

Preparation and the in-vivo evaluation of paclitaxel liposomes for lung targeting delivery in dogs

Objectives The aim of this study was to develop paclitaxel liposomes for a lung targeting delivery system. Methods The liposomes composed of Tween‐80/HSPC/cholesterol (0.03 : 3.84 : 3.84, mol/mol), containing paclitaxel and lipids (1 : 40, mol/mol), were prepared by a combination of solid dispersion and effervescent techniques, and then subjected to ultrasonication. The pharmacokinetics and biodistribution of liposomal and injectable formulation of paclitaxel in dogs were studied after intravenous administration. Key findings The mean diameter, polydispersity index, zeta‐potential and entrapment efficiency of the liposomes were 501.60 ± 15.43 nm, 0.28 ± 0.02, ?20.93 ± 0.06 mV and 95.17 ± 0.32%, respectively. The liposomal formulation kept stable for at least 3 months at 6 ± 2°C and didn't cause haemolysis. The liposome carrier decreased the area under the curve and terminal half‐life of paclitaxel compared with paclitaxel injection ranging from 0.352 ± 0.031 mg/l*h and 0.0671 ± 0.144 h to 0.748 ± 0.062 mg/l*h and 1.978 ± 0.518 h, respectively. The paclitaxel liposomes produced a drug concentration in the lung that was markedly higher than that in other organs or tissues and was about 15‐fold of that of paclitaxel injection at 2 h. Conclusions To sum up, these results demonstrated that the paclitaxel liposomes are an effective lung targeted carrier in the treatment of lung cancer.     

Liposomes Modified with Cyclic RGD Peptide for Tumor Targeting

Cyclic RGD peptide anchored sterically stabilized liposomes (RGD-SL) were investigated for selective and preferential presentation of carrier contents at angiogenic endothelial cells overexpressing α_vβ₃ integrins on and around tumor tissue and thus for assessing their targetabilty. Liposomes were prepared using distearoylphosphatidylcholine (DSPC), cholesterol and distearoylphosphatidylethanolamine–polyethyleneglycol–RGD peptide conjugate (DSPE–PEG–RGD) in a molar ratio 56:39:5. The control RAD peptide anchored sterically stabilized liposomes (RAD-SL) and liposome with 5 mol% PEG (SL) without peptide conjugate which had similar lipid composition were used for comparison. The average size of all liposome preparations prepared was approximately 105 nm and maximum drug entrapment was 10.2±1.1%. In vitro endothelial cell binding of liposomes exhibited 7-fold higher binding of RGD-SL to HUVEC in comparison to the SL and RAD-SL. Spontaneous lung metastasis and angiogenesis assays show that RGD peptide anchored liposomes are significantly (p<0.01) effective in the prevention of lung metastasis and angiogenesis compared to free 5-FU, SL and RAD-SL. In therapeutic experiments, 5-FU, SL, RGD-SL and RAD-SL were administered intravenously on day 4 at the dose of 10 mg 5-FU/kg body weight to B16F10 tumor bearing BALB/c mice resulting in effective regression of tumors compared with free 5-FU, SL and RAD-SL. Results indicate that cyclic RGD peptide anchored sterically stabilized liposomes bearing 5-FU are significantly (p<0.01) active against primary tumor and metastasis than the non-targeted sterically stabilized liposomes and free drug. Thus cyclic RGD peptide anchored sterically stabilized liposomes hold potential of targeted cancer chemotherapeutics.     

Polysaccharide-modified liposomes and their application in cancer research

Nanodrug delivery systems have been widely used in cancer treatment. Among these, liposomal drug carriers have gained considerable attention due to their biocompatibility, biodegradability, and low toxicity. However, conventional liposomes have several shortcomings, such as poor stability, rapid clearance, aggregation, fusion, degradation, hydrolysis, and oxidation of phospholipids. Polysaccharides are natural polymers of biological origin that exhibit structural stability, excellent biocompatibility and biodegradability, flexibility, non-immunogenicity, low toxicity, and targetability. Therefore, they represent a promising class of polymers for the modification of the surface properties of liposomes to overcome their shortcomings. In addition, polysaccharides can be readily combined with other materials to develop new composite materials. Hence, they represent the optimal choice for liposomal modification to improve pharmacokinetics and clinical utility. Polysaccharide-coated liposomes exhibit better stability, drug release kinetics, and cellular uptake than conventional liposomes. The oncologic application of polysaccharide-coated liposomes has become a research hotspot. We summarize the preparation, physicochemical properties, and antineoplastic effects of polysaccharide-coated liposomes to facilitate antitumor drug development.     

注射用脂质体紫杉醇抗F344大鼠卵巢癌移植瘤的作用

目的:探讨注射用脂质体紫杉醇(力朴素)对F344大鼠卵巢癌移植瘤抗肿瘤活性的影响。方法:采用NuTu-19卵巢癌细胞对F344大鼠建立卵巢癌移植瘤大鼠模型,分别用紫杉醇和力朴素(5 mg·kg^-1)进行腹腔注射治疗。采用量筒对腹水进行测量;对肿瘤进行称重;ELISA法检测CK,AST和LDH的含量;HE染色法对肿瘤组织进行病理观察。结果:与紫衫醇相比,力朴素对荷瘤F344大鼠的腹水体积及肿瘤质量无明显差别,CK,AST,LDH以及CA-125显著降低,病理显示肿瘤细胞生长被抑制。结论:同等剂量下,力朴素具有与紫杉醇相似的抗肿瘤活性,副作用低。     

Increasing the antitumor efficacy of doxorubicin-loaded liposomes with peptides anchored via a chelator lipid

The therapeutic efficacy of anticancer drugs like doxorubicin can be significantly increased by their incorporation into liposomes, but an ability to actively target the drug-containing liposomes to tumors could well provide an even greater curative effect. In this work, a commercial preparation of doxorubicin-loaded liposomes (Caelyx) was modified by incorporation of the metal chelator lipid 3(nitrilotriacetic acid)-ditetradecylamine (NTA₃-DTDA) to enable engraftment of histidine-tagged targeting molecules. Our results show that when engrafted with p15-RGR, a His-tagged peptide containing a sequence purported to bind platelet-derived growth factor receptor β (PDGFRβ), NTA₃-DTDA-containing Caelyx (3NTA-Caelyx) can be targeted to NIH-3T3 cells in vitro, leading to increased cytotoxicity compared with non-targeted 3NTA-Caelyx. PDGFRβ is known to be expressed on pericytes in the tumor vasculature; however, when radiolabeled p15-RGR liposomes were administered to mice bearing subcutaneous B16-F1 tumors, minimal accumulation into tumors was observed. In contrast, an alternative targeting peptide, p46-RGD, was found to actively direct liposomes to tumors (4.7 %ID/g). Importantly, when injected into tumor-bearing mice, p46-RGD-engrafted 3NTA-Caelyx significantly decreased the tumor growth rate compared with controls. These results indicate that the incorporation of NTA₃-DTDA into liposomal drugs could represent a simple modification to the drug to allow engraftment of targeting molecules and to increase its efficacy.     

紫杉醇壳聚糖聚合物胶束的制备及表面电荷对其在小鼠体内组织分布的影响

目的紫杉醇壳聚糖聚合物胶束的制备及表面电荷对其在小鼠体内组织分布的影响。方法采用透析法分别制备紫杉醇阳离子(PTX-CCM)和阴离子(PTX-ACM)壳聚糖聚合物胶束；昆明种小鼠分别尾静脉注射20 mg·kg^-1的PTX-CCM和PTX-ACM，HPLC法测定各组织中不同时间的药物含量，以各组织药代动力学参数(AUC，MRT，T_max和C_max)评价其体内分布。结果PTX-CCM和PTX-ACM粒径分别为164和180 nm，zeta电位分别为+23.7和-28.0 mV，载药量分别为26.4%和34.6%(w/w)，包封率分别为76.2%和89.9%。PTX-CCM和PTX-ACM肝中最大药物分布量分别达给药量的64.72%和91.84%，MRT分别为5.50和51.39 h；PTX-CCM和PTX-ACM脾中最大药物分布量达给药量的7.08%和5.16%，MRT分别为9.04和26.82 h；PTX-CCM肺部AUC和C_max分别为PTX-ACM的2.71和5.87倍；PTX-CCM和PTX-ACM心脏最高药物分布量仅为给药量的0.36%和0.24%，肾脏仅为给药量的0.75%和0.33%。结论PTX-CCM和PTX-ACM表面分别带正负电荷，具有优良的载药性能，两者皆显示出对肝脾的高度亲和性和滞留特性，尤以PTX-ACM更为显著；PTX-CCM较PTX-ACM具有更好的肺靶向性，但肺部滞留性相对较弱；两者在心、肾的分布均极少，可有效降低PTX对这些器官的毒副作用。     

Transferrin and octaarginine modified dual-functional liposomes with improved cancer cell targeting and enhanced intracellular delivery for the treatment of ovarian cancer

Off-target effects of drugs severely limit cancer therapy. Targeted nanocarriers are promising to enhance the delivery of therapeutics to tumors. Among many approaches for active tumor-targeting, arginine-rich cell penetrating peptides (AR-CPP) and ligands specific to target over-expressed receptors on cancer-cell surfaces, are popular. Earlier, we showed that the attachment of an AR-CPP octaarginine (R8) to the surface of DOXIL^® (Doxorubicin encapsulated PEGylated liposomes) improved cytoplasmic and nuclear DOX delivery that enhanced the cytotoxic effect in vitro and improved therapeutic efficacy in vivo. Here, we report on DOX-loaded liposomes, surface-modified with, R8 and transferrin (Tf) (Dual DOX-L), to improve targeting of A2780 ovarian carcinoma cells via the over-expressed transferrin receptors (TfRs) with R8-mediated intracellular DOX delivery. Flow cytometry analysis with fluorescently labeled DualL (without DOX) showed two-fold higher cancer-cell association than other treatments after 4?h treatment. Blocking entry pathways of R8 (macropinocytosis) and Tf (receptor-mediated endocytosis, RME) resulted in a decreased cancer-cell association of DualL. Confocal microscopy confirmed involvement of both entry pathways and cytoplasmic liposome accumulation with nuclear DOX delivery for Dual DOX-L. Dual DOX-L exhibited enhanced cytotoxicity in vitro and was most effective in controlling tumor growth in vivo in an A2780 ovarian xenograft model compared to other treatments. A pilot biodistribution study showed improved DOX accumulation in tumors after Dual DOX-L treatment. All results collectively presented a clear advantage of the R8 and Tf combination to elevate the therapeutic potential of DOX-L by exploiting TfR over-expression imparting specificity followed by endosomal escape and intracellular delivery via R8.     

结肠癌靶向米托蒽醌脂质体的制备及其抗肿瘤活性研究

目的:比较透明质酸(HA)修饰的脂质体(HA-LP)与普通脂质体(LP)的抗转移及抗肿瘤活性。方法:采用薄膜水化法制备空白脂质体,通过透明质酸上的羧基与膜材中DSPE的氨基反应将HA连接在脂质体上,运用硫酸铵梯度法包载模型药物米托蒽醌;通过透射电镜表征脂质体形态;免疫荧光法检测结肠癌细胞(CT26 cells)中透明质酸受体CD44表达情况;高内涵扫描仪检测结肠癌细胞对脂质体的摄取情况;划痕实验考察2组脂质体抑制细胞转移的能力;体内抗肿瘤实验考察2组脂质体的抑瘤效果。结果:所制备的脂质体形态均一,包封率均大于95%。免疫荧光结果显示,结肠癌细胞中CD44受体高度表达。相同的药物浓度下,HA-LP相较于未修饰的脂质体具有明显增加的细胞摄取。划痕实验结果表明,HA可能与受体CD44存在某种相互作用,抑制了结肠癌细胞的转移,表现出HA-LP具有更强的抗转移活性。体内抗肿瘤结果显示,HA-LP对小鼠荷结肠癌实体瘤的抑瘤率为62.1%,显著高于普通脂质体组(46.6%,P<0.05)。结论:HA-LP与细胞膜上的受体CD44作用,显著增加了其细胞摄取,并抑制了结肠癌细胞的转移,体内抑瘤结果同样显示,具有肿瘤靶向功能的HA-LP具有更强的抑制肿瘤生长的能力。     

Carrier-based strategies for targeting protein and peptide drugs to the lungs

With greater interest in delivery of protein and peptide-based drugs to the lungs for topical and systemic activity, a range of new devices and formulations are being investigated. While a great deal of recent research has focused on the development of novel devices, attention must now be paid to the formulation of these macromolecular drugs. The emphasis in this review will be on targeting of protein/peptide drugs by inhalation using carriers and ligands.     

紫杉醇纳米脂质体的制备与大鼠体内药动学

目的:制备紫杉醇新型纳米脂质体并研究其在大鼠体内的药动学。方法:采用薄膜分散超声结合冷冻干燥制备紫杉醇纳米脂质体。大鼠尾静脉注射紫杉醇脂质体及市售紫杉醇注射液Anzatax,血浆样品经乙醚提取后用反相高效液相色谱法(RP-HPLC)检测血浆紫杉醇浓度,并用3P87软件包估算药动学参数。色谱条件如下:色谱柱为Gemini ODS(150 mm×4.6 mm,5μm),流动相为0．035mol·L-1乙酸铵缓冲液(pH 5．0)-乙腈(45:50),流速1．0 mL·min,检测波长230 nm,地西泮为内标。结果:制备的紫杉醇纳米脂质体的体积权重粒径为(54.1±26．0)nm,包封率大于80％,符合药典规定,且24 h内与葡萄糖注射液配伍稳定。紫杉醇脂质体及市售紫杉醇注射液Anzatax经大鼠尾静脉注射后均符合二室模型,脂质体组的消除半衰期(t1/2β)显著长于Anzatax组[(3．38±0．39)vs．(2．49±0．63)h,P<0．05];其他药动学参数经方差分析均无显著性差异。紫杉醇脂质体与Anzatax的AUC0-8h比值为88．13％。结论:制备的紫杉醇纳米脂质体在大鼠体内的药动物参数与市售紫杉醇注射液Anzatax比较,t1/2β稍有延长,AUC0-8h值相近。     

抗菌肽的抗肿瘤作用研究新进展

抗菌肽是一类广泛存在于生物体内的天然小分子多肽,它是机体免疫系统的天然组分,具有抗菌、抗炎及抗病毒等作用.许多抗菌肽具有抗肿瘤活性,而且在细胞选择性和耐药性方面也拥有独特的优势.本文综述抗菌肽的抗肿瘤机制及应用.     

消癌平注射液联合紫杉醇对人卵巢癌SK-OV-3细胞增殖及裸鼠异位移植瘤生长的作用

目的 研究消癌平（XAP）注射液和紫杉醇（PTX）联合应用对人卵巢癌SK-OV-3细胞增殖及裸鼠异位移植瘤的抑制作用。方法 将生长至对数期的人卵巢癌SK-OV-3细胞分为6组：对照组、紫杉醇组（10 nmol/L）、消癌平注射液低、高剂量组（20、80 μl/ml）、消癌平注射液低、高剂量联合组（PTX 10 nmol/L+XAP 20 μl/ml、PTX 10 nmol/L+XAP 80 μl/ml）,各组分别加入药物干预24 h或48 h后,MTT法测定并计算细胞存活率。将36只雌性荷人卵巢癌细胞SK-OV-3的异位移植瘤裸鼠随机分为6组,包括对照组（G1：生理盐水）、紫杉醇组（G2：PTX 10 mg/kg）、消癌平注射液低、高剂量组（G3：XAP 20 ml/kg、G4：XAP 50 ml/kg）、低剂量联合组（G5：PTX 10 mg/kg+XAP 20 ml/kg）以及高剂量联合组（G6：PTX 10 mg/kg+XAP 50 ml/kg）,每组6只。各治疗组予相应药物治疗18 d,观察记录小鼠一般情况、体重、肿瘤体积,并计算抑瘤率。结果 SK-OV-3细胞体外实验结果显示,与对照组相比,消癌平注射液、低剂量联合组和高剂量联合组的SK-OV-3细胞存活率均显著降低（P<0.05或P<0.01）,且联合用药组与紫杉醇组相比,也有显著差异（P<0.05）,并呈现剂量和时间依赖性。裸鼠异位移植瘤实验结果显示,高剂量联合组的抑瘤率明显高于对照组和紫杉醇组,差异有统计学意义（P<0.05）。结论 消癌平注射液和紫杉醇联用对人卵巢癌细胞SK-OV-3裸鼠异位移植瘤的抑制具有协同作用（P<0.05）。     

肺靶向阿奇霉素脂质体的制备及其在小鼠体内的分布

目的研究肺靶向阿奇霉素阳离子脂质体的制备方法并考察其在小鼠体内的分布。方法利用旋转薄膜-冻融法制备肺靶向阿奇霉素脂质体。用高效液相色谱法测定给药后小鼠体内各组织中的药物浓度。结果制得的脂质体平均粒径为6.582 μm，表面电荷为+19.5 mV，包封率大于75%，稳定性好。药物体外释药符合Higuchi方程。小鼠尾静脉给药后，阳离子脂质体主要被肺摄取，在肺部的滞留时间明显延长，AUC值约为阿奇霉素溶液的8.4倍。结论采用薄膜-冻融法，添加十八胺可制得具有较高包封率及稳定性的阿奇霉素阳离子脂质体，在小鼠肺部的分布优于注射液，能达到肺靶向目的。     

Estrogen-functionalized liposomes grafted with glutathione-responsive sheddable chotooligosaccharides for the therapy of osteosarcoma

An estrogen (ES)-functionalized cationic liposomal system was developed and exploited for targeted delivery to osteosarcoma. Natural biocompatible chotooligosaccharides (COS, MW2-5?KDa) were covalently tethered to the liposomal surface through a disulfate bond (-SS-) to confer reduction-responsive COS detachment, whereas estrogen was grafted via polyethylene glycol (PEG 2?K) chain to achieve estrogen receptor-targeting. The liposomal carriers were prepared by the ethanol injection method and fluorescent anticancer drug doxorubicin (DOX) was loaded with ammonium sulfate gradient. The physicochemical properties, reduction-sensitivity, and the roles of estrogen on cellular uptake and tumor-targeting were studied. The Chol-SS-COS/ES/DOX liposomes were spherical with an average size about 110?nm, and high encapsulation efficiency. The liposomes were stable in physiological condition but rapidly release the payload in response to tumoral intracellular glutathione (20?mM). MTT cytotoxicity assay confirmed that Chol-SS-COS/ES/DOX liposomes exhibited higher cytotoxicity to MG63 osteosarcoma cells than to liver cells (LO2). Flow cytometry (FCM) and confocal laser scanning microscopy revealed that cellular uptake of Chol-SS-COS/ES/DOX liposomes by MG63, than the free DOX or Chol-SS-COS/DOX. Ex vivo fluorescence distribution study showed that the multifunctional liposomes selectively accumulated in the MG63 xenografts versus the organs. Chol-SS-COS/ES/DOX liposomes strongly inhibited the tumor growth and enhanced the animal survival rate. Overall, the COS grafted estrogen-functionalized cationic liposomes, fortified with glutathione-responsiveness, showed great potential for specific intracellular drug delivery to estrogen receptor-expressing tumors such as osteosarcoma.