Chemotherapy with Hybrid Liposomes without Any Drug &lt;i&gt;in Vivo&lt;/i&gt;

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Duchenne/Becker muscular dystrophy (DMD/BMD) is caused by a defective expression of the dystrophin gene resulting in the absence of the dystrophin protein in muscle fibers.1,2) Approximately 60% of DMD/BMD patients have deletions in the dystrophin gene itself, [3][4][5] while the remaining 40% have small deletions or point mutations in the region that encodes the gene. Furthermore, nonsense mutations located within the gene account for approximately 5-13% of the muscular dystrophies. 6,7)Aminoglycoside antibiotics such as gentamicin (GM) had the ability to allow the ribosome to read through a premature-termination codon of the dystrophin gene, which prevented normal translation of dystrophin protein.8,9) BartonDavis et al. demonstrated the possibility of treating X chromosome-linked muscular distrophy (mdx) mouse, which was an animal model for DMD that possessed a nonsense mutation in the dystrophin gene, with GM in vivo. 10) They used GM to suppress the nonsense mutations and could restore dystrophin expression successfully in mdx mouse. However, the GM therapy has been hindered by several problems such as severe side effects of GM, especially nephrotoxicity and ototoxicity, the poor delivery profile to muscle tissue, and short half-life in blood. Recently, the phase 2b clinical trial of PTC 124 (3-[5-(2-fluorophenyl)-[1,2,4]oxadiazol-3-yl]-benzoic acid), 11) which is a new drug to induce reading through a premature-termination codon without clear side-effects, showed that the primary endpoint of the change in 6 min walk distance tests did not reach any statistical significance within the 48 weeks duration of the study according to Genzyme corporation announcement. 12)Therefore, to overcome these inadequacies of GM therapy for DMD, we encapsulated GM in hybrid liposomes (HL) for the delivery system. HL can be prepared by just the sonication of vesicular and micellar molecules in a buffer solution.13,14) HL are free from any contamination with organic solvents and remain stable for longer periods. The physical properties of these liposomes such as size, membrane fluidity, phase transition temperature, and hydrophobicity can be controlled by changing the constituents and compositional ratios.In the course of our study for HL, the following interesting results have been obtained. 23) and hydrophobic agents in the drug delivery system (DDS).In this study, we reported the therapeutic effects of HL including GM (GM-HL) on the mdx mice in vivo. The reduction of side effects of GM-HL is also discussed on the basis of the results from auditory brainstem response (ABR) tests and biodistribution analysis of HL. Kumamoto 860-0811, Japan. Received December 14, 2010; accepted January 28, 2011; published online February 7, 2011 It is known that gentamicin (GM) could be a possible treatment for Duchenne Muscular Dystrophy (DMD). However, GM therapy has been hindered by several problems such as severe side effects of GM. In order to resolve these problems, we developed the drug delivery system (DDS) of GM using hybrid lip...

Section: Discussionmentioning

confidence: 93%

Effective Drug Delivery System for Duchenne Muscular Dystrophy Using Hybrid Liposomes Including Gentamicin along with Reduced Toxicity

Yukihara

Ito

Tanoue

et al. 2011

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“…We have already reported that HL and cationic HL have no side-effects in vivo in the safety tests intravenously administered via a vein using mice 9,21) and rats. [17][18][19] These results indicate that cationic HL could not have severe side effects.…”

Section: Autopsy Analysis Of the Hepatic Metastasismentioning

confidence: 85%

“…16 21 , 65.7 mg/kg/d for 2C 14 ECl) were intravenously administered once each day for 14 d. Safety of 136 mg/kg/d for DMPC in chronic toxicity test using mice and rats has been reported. [17][18][19] After 14 d, the livers were isolated from the mouse models, fixed in 10% formalin solution, embedded in paraffin and sectioned at 5 µm of thickness. The liver sections were stained with hematoxylin and eosin (H&E) and observed by an optical microscope (Nikon TS-100, Tokyo, Japan).…”

Section: Preparation Of Hybrid Liposomesmentioning

confidence: 99%

Therapeutic Effects of Cationic Hybrid Liposomes on the Hepatic Metastasis of Colon Carcinoma Along with Apoptosis <i>in Vivo</i>

Ichihara

Hino

Ueoka

et al. 2014

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Therapeutic effects of cationic hybrid liposomes (HL) composed of 87 mol% dimyristoyl-phosphatidylcholine (DMPC), 5 mol% polyoxyethylene (21) dodecyl ether (C 12 (EO) 21 ) and 8 mol% O,O′-ditetradecanoyl-N-(α-trimethyl-ammonioacetyl) diethanolamine chloride (2C 14 ECl) on the metastasis of human colon carcinoma (HCT116) cells were examined in vivo. Cationic HL having a hydrodynamic diameter less than 150 nm were preserved for one month. Therapeutic effects were obtained in the hepatic metastasis mouse models of HCT116 cells after the intravenous injection of cationic HL. The histological analysis indicated the induction of apoptosis in the liver section of the hepatic metastasis mouse models treated with cationic HL in vivo. Therapeutic effects of cationic HL without any drugs on the hepatic metastasis were revealed for the first time on the basis of histological analyses in vivo.Key words hybrid liposome; hepetic metastasis; colon carcinoma; apoptosis; chemotherapy Colorectal cancer (CRC) is the third most common cancer in the world with more than 600000 deaths every year. 1)About 40% of patients with CRC develop metastases. Because the venous drainage of the colon is through the portal vein, which goes directly to the liver, more than 70% of the CRC metastases are located in the hepatic tissue. Metastatic diffusion results in a very poor prognosis with a median survival of about two years in treated patients. However, long term survival is possible in the 15% of patients that can benefit of hepatic metastasis surgery, usually after induction chemotherapy.2) Induction chemotherapy is a first-line treatment for cancer in which a patient is given doses of chemotherapy first. These doses may be high, with the goal of attempting to quickly attack the cancer, and after induction chemotherapy, additional treatment options such as surgical resection can be selected.3) While the chemotherapy drugs kill tumor cells, they also damage normal cells, causing severe side-effects such as gastrointestinal dysfunction and bone marrow toxicity. Therefore, chemotherapy that is effective for the metastasis of cancers without any side-effects is required.On the other hand, we have produced hybrid liposomes (HL) which can be prepared by just the sonication of vesicular and micellar molecules in a buffer solution. 4,5) HL are free from any contamination with organic solvents and remain stable for longer periods. The physical properties of these liposomes such as size, membrane fluidity, phase transition temperature, and hydrophobicity can be controlled by changing the constituents and compositional ratios of the HL. In the course of our study for HL, the following interesting results have been obtained. (a) Stereochemical control of the enantioselective hydrolysis of amino acid esters could be established by temperature regulation and changing the composition of the HL. 14) However, therapeutic effects of cationic HL in vivo have not yet been examined.In this study, we investigated the therapeutic effects of three-component cationic ...

“…9) (d) No toxicity of the hybrid liposomes was observed in normal cells in vitro nor in normal rats in vivo. 8,10) In this study, we report on hybrid liposomes composed of L-a -dimyristoylphosphatidylcholine (DMPC) and polyoxyethylenedodecyl ether (C 12 (EO) n : CH 3 (CH 2 ) 11 O(CH 2 CH 2 O) n H) having inhibitory effects on the growth of human breast tumor cells (MDA-MB-453) in vitro along with apoptosis.Hybrid liposomes were prepared by sonication (VELVO VS-N300, 300W) of a mixture containing 95 mol% DMPC and 5 mol% C 12 (EO) n in 5% glucose solution at 45°C with 300W, followed by filtration with a 0.20 mm filter.First, we examined the fifty percent inhibitory concentration (IC 50 ) of hybrid liposomes (HL-n) on the growth of MDA-MB-453 cells in vitro on the basis of WST-1 assay. …”

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confidence: 99%

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confidence: 99%

Induction of Apoptosis by Hybrid Liposomes for Human Breast Tumor Cells along with Activation of Caspases

Nagami

Matsumoto

Ueoka

2006

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It is well known that liposomes are used as drug carriers: examples are antitumor agents, hormones, and immunomodulation. 1,2) On the other hand, we have recently produced specific hybrid liposomes composed of vesicular and micellar molecules; they are stable for a longer period. The physical properties of these liposomes such as size, membrane fluidity, phase transition temperature, and hydrophobicity can be controlled by changing the constituents and compositional ratios of hybrid liposomes. 3,4) In the course of our study on hybrid liposomes, the following interesting results were obtained. (a) Stereochemical control of the enantioselective hydrolysis of amino acid esters could be established by temperature regulation and by changing the composition of hybrid liposomes. 9) (d) No toxicity of the hybrid liposomes was observed in normal cells in vitro nor in normal rats in vivo. 8,10) In this study, we report on hybrid liposomes composed of L-a -dimyristoylphosphatidylcholine (DMPC) and polyoxyethylenedodecyl ether (C 12 (EO) n : CH 3 (CH 2 ) 11 O(CH 2 CH 2 O) n H) having inhibitory effects on the growth of human breast tumor cells (MDA-MB-453) in vitro along with apoptosis.Hybrid liposomes were prepared by sonication (VELVO VS-N300, 300W) of a mixture containing 95 mol% DMPC and 5 mol% C 12 (EO) n in 5% glucose solution at 45°C with 300W, followed by filtration with a 0.20 mm filter.First, we examined the fifty percent inhibitory concentration (IC 50 ) of hybrid liposomes (HL-n) on the growth of MDA-MB-453 cells in vitro on the basis of WST-1 assay. 11)The tumor cells (5.0ϫ10 4 viable cells/ml) were cultured for 48 h in an incubator at 37°C after adding the sample solutions. WST-1 solutions were added and the absorbance at wavelength of 450 nm was measured by spectrophotometer. The inhibitory concentration was evaluated by A mean / A control , where A mean and A control denote the absorbance of water-soluble formazan, which was useful as an indicator of cell viability, in the presence and absence of sample solutions, respectively. The results are shown in Fig. 1. The IC 50 values of HL-n were from one seventh to a half of those of the DMPC liposomes, indicating that the inhibitory effects of hybrid liposomes are large when compared with those of the DMPC liposomes.Morphology of hybrid liposomes (HL-n) was examined on the basis of dynamic light scattering measurements. Hydrodynamic diameter (d hy ) of HL-n was 80-120 (HL-21 and 23) and 90-110 nm (HL-25), which remained stable for more than three weeks. On the other hand, it was found that DMPC liposomes, HL-4 and HL-10 were unstable. It is worthy to note that HL-n (nϭ21, 23, 25) having a diameter of 100 nm could avoid the clearance by reticular endothelial system in vivo. 12)Second, we examined the mechanism for inhibiting the growth of MDA-MB-453 cells in vitro. Fluorescence micrographs of MDA-MB-453 cells using TUNEL method after the treatment with hybrid liposomes are shown in Fig. 2. The green color (FITC) was observed in the cells after adding hybrid lipos...