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Nano-Drugs Based on Nano Sterically Stabilized Liposomes for the Treatment of Inflammatory Neurodegenerative Diseases.

Turjeman K, Bavli Y, Kizelsztein P, Schilt Y, Allon N, Katzir TB, Sasson E, Raviv U, Ovadia H, Barenholz Y - PLoS ONE (2015)

Bottom Line: For the NSSL-MPS we also compared the effect of passive targeting alone and of active targeting based on short peptide fragments of ApoE or of β-amyloid.Our results clearly show that for NSSL-MPS, active targeting is not superior to passive targeting.The highly efficacious anti-inflammatory therapeutic feature of these two nano-drugs meets the criteria of disease-modifying drugs and supports further development and evaluation of these nano-drugs as potential therapeutic agents for diseases with an inflammatory component.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Membrane and Liposome Research, Department of Biochemistry and Molecular Biology, Institute for Medical Research Israel-Canada (IMRIC), The Hebrew University-Hadassah Medical School, Jerusalem, Israel.

ABSTRACT
The present study shows the advantages of liposome-based nano-drugs as a novel strategy of delivering active pharmaceutical ingredients for treatment of neurodegenerative diseases that involve neuroinflammation. We used the most common animal model for multiple sclerosis (MS), mice experimental autoimmune encephalomyelitis (EAE). The main challenges to overcome are the drugs' unfavorable pharmacokinetics and biodistribution, which result in inadequate therapeutic efficacy and in drug toxicity (due to high and repeated dosage). We designed two different liposomal nano-drugs, i.e., nano sterically stabilized liposomes (NSSL), remote loaded with: (a) a "water-soluble" amphipathic weak acid glucocorticosteroid prodrug, methylprednisolone hemisuccinate (MPS) or (b) the amphipathic weak base nitroxide, Tempamine (TMN). For the NSSL-MPS we also compared the effect of passive targeting alone and of active targeting based on short peptide fragments of ApoE or of β-amyloid. Our results clearly show that for NSSL-MPS, active targeting is not superior to passive targeting. For the NSSL-MPS and the NSSL-TMN it was demonstrated that these nano-drugs ameliorate the clinical signs and the pathology of EAE. We have further investigated the MPS nano-drug's therapeutic efficacy and its mechanism of action in both the acute and the adoptive transfer EAE models, as well as optimizing the perfomance of the TMN nano-drug. The highly efficacious anti-inflammatory therapeutic feature of these two nano-drugs meets the criteria of disease-modifying drugs and supports further development and evaluation of these nano-drugs as potential therapeutic agents for diseases with an inflammatory component.

No MeSH data available.


Related in: MedlinePlus

Comparison of passively targeted NSSL and actively targeted peptide-conjugated NSSL.(A) Representative fluorescent microscopy images comparing brain accumulation of NSSL and their payload as is (A, A1), β-amyloid NSSL(B,B1), and ApoE NSSL (C,C1) in healthy mice brain showing an increase in the amount of actively targeted NSSL and their payload accumulating, compared to passively targeted NSSL. (B) Comparison of the therapeutic efficacy of passively targeted NSSL-MPS and actively targeted peptide-conjugated NSSL-MPS in the acute EAE mice model. SJL mice were treated by IV injections on days 10, 12, 14 post-immunization with saline (control) (◆), NSSL-MPS (●), Apo-E NSSL-MPS (▲) or β-amyloid NSSL-MPS (■). * p-value < 0.0001.
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pone.0130442.g006: Comparison of passively targeted NSSL and actively targeted peptide-conjugated NSSL.(A) Representative fluorescent microscopy images comparing brain accumulation of NSSL and their payload as is (A, A1), β-amyloid NSSL(B,B1), and ApoE NSSL (C,C1) in healthy mice brain showing an increase in the amount of actively targeted NSSL and their payload accumulating, compared to passively targeted NSSL. (B) Comparison of the therapeutic efficacy of passively targeted NSSL-MPS and actively targeted peptide-conjugated NSSL-MPS in the acute EAE mice model. SJL mice were treated by IV injections on days 10, 12, 14 post-immunization with saline (control) (◆), NSSL-MPS (●), Apo-E NSSL-MPS (▲) or β-amyloid NSSL-MPS (■). * p-value < 0.0001.

Mentions: To answer the question whether treatment with actively targeted NSSL-MPS (as opposed to the passively targeted NSSL-MPS) will further improve the therapeutic efficacy of these drugs we fabricated actively targeted NSSL-MPS, presenting peptides that enable them to cross the blood-brain barrier and release their payload in the brain. As the targeting ligands we used short peptide fragments of ApoE or of β-amyloid that were pre-conjugated to dioleoyl succinate (DO-succinate). Preliminary results using NSSL liposomes labeled with Texas Red and loaded with calcein showed increases in the amount of actively targeted liposomes and their payload accumulated in healthy mice brains after injecting these mice with actively targeted liposomes, compared to passively targeted liposomes. Fig 6A shows representative fluorescent microscopy images comparing brain accumulation of NSSL and their payload: NSSL as is (A, A1); β-amyloid NSSL (B,B1); and ApoE NSSL (C,C1) in healthy mice brain showing an increase in the amount of actively targeted NSSL and their payload accumulated compared to passively targeted NSSL. We further compared the therapeutic efficacy of these targeted NSSL loaded with MPS in the acute EAE mice model (Fig 6B, Table 6). SJL mice were injected on days 10, 12, 14 post immunization with saline (control) or 10mg/kg NSSL-MPS (with or without the targeting peptides attached to their surface). Gaillard et al. [57] tried to enhance brain penetration of NSSL-MPS by using a targeted NSSL-MPS formulation (based on our previously published remote loaded NSSL-MPS formulation conjugated to the brain-targeting ligand glutathione), demonstrating a slight advantage (at best) in the MBP-EAE rats over the passively targeted formulation, apparent only 24 h after treatment. In the present study, although at the end of the experiment, a week after the third and last injection, MPS levels were significantly higher in the targeted liposomes-treated groups compared to the NSSL-MPS-treated group (Fig 6B). Both targeted formulations (ApoE- and β-amyloid-conjugated NSSL) did not show any therapeutic advantages over the passively targeted NSSL formulation, although all three formulations significantly ameliorated disease severity compared to control group. Furthermore, β-amyloid NSSL-MPS treatment was significantly less efficacious than passively targeted NSSL-MPS treatment. Treatment with ApoE NSSL-MPS showed similar therapeutic efficacy to treatment with NSSL-MPS (mean burden of disease: 0.9±0.1 and 1±0.1, respectively) compared to control (mean burden of disease 2.4±0.17). Treatment with β-amyloid NSSL-MPS lowered slightly the mean maximal clinical scores (3.2±0.31) and the mean burden of disease (1.7±0.14) compared to the control group (3.9±0.35, 2.4±0.17). Therefore, the improved penetration into the brain did not correlate with improved efficacy. The actively targeted NSSL probably distribute less uniformly throughout the diseased brains while remaining attached to cells close to the blood vessels, similarly to what was demonstrated before for tumor targeted NSSL [58]. In addition, BBB disruption in EAE mice, which enables penetration of passively targeted NSSL, might explain the lack of therapeutic advantage of actively targeted NSSL which may be able to extravasate the intact BBB over the passively targeted NSSL.


Nano-Drugs Based on Nano Sterically Stabilized Liposomes for the Treatment of Inflammatory Neurodegenerative Diseases.

Turjeman K, Bavli Y, Kizelsztein P, Schilt Y, Allon N, Katzir TB, Sasson E, Raviv U, Ovadia H, Barenholz Y - PLoS ONE (2015)

Comparison of passively targeted NSSL and actively targeted peptide-conjugated NSSL.(A) Representative fluorescent microscopy images comparing brain accumulation of NSSL and their payload as is (A, A1), β-amyloid NSSL(B,B1), and ApoE NSSL (C,C1) in healthy mice brain showing an increase in the amount of actively targeted NSSL and their payload accumulating, compared to passively targeted NSSL. (B) Comparison of the therapeutic efficacy of passively targeted NSSL-MPS and actively targeted peptide-conjugated NSSL-MPS in the acute EAE mice model. SJL mice were treated by IV injections on days 10, 12, 14 post-immunization with saline (control) (◆), NSSL-MPS (●), Apo-E NSSL-MPS (▲) or β-amyloid NSSL-MPS (■). * p-value < 0.0001.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4492950&req=5

pone.0130442.g006: Comparison of passively targeted NSSL and actively targeted peptide-conjugated NSSL.(A) Representative fluorescent microscopy images comparing brain accumulation of NSSL and their payload as is (A, A1), β-amyloid NSSL(B,B1), and ApoE NSSL (C,C1) in healthy mice brain showing an increase in the amount of actively targeted NSSL and their payload accumulating, compared to passively targeted NSSL. (B) Comparison of the therapeutic efficacy of passively targeted NSSL-MPS and actively targeted peptide-conjugated NSSL-MPS in the acute EAE mice model. SJL mice were treated by IV injections on days 10, 12, 14 post-immunization with saline (control) (◆), NSSL-MPS (●), Apo-E NSSL-MPS (▲) or β-amyloid NSSL-MPS (■). * p-value < 0.0001.
Mentions: To answer the question whether treatment with actively targeted NSSL-MPS (as opposed to the passively targeted NSSL-MPS) will further improve the therapeutic efficacy of these drugs we fabricated actively targeted NSSL-MPS, presenting peptides that enable them to cross the blood-brain barrier and release their payload in the brain. As the targeting ligands we used short peptide fragments of ApoE or of β-amyloid that were pre-conjugated to dioleoyl succinate (DO-succinate). Preliminary results using NSSL liposomes labeled with Texas Red and loaded with calcein showed increases in the amount of actively targeted liposomes and their payload accumulated in healthy mice brains after injecting these mice with actively targeted liposomes, compared to passively targeted liposomes. Fig 6A shows representative fluorescent microscopy images comparing brain accumulation of NSSL and their payload: NSSL as is (A, A1); β-amyloid NSSL (B,B1); and ApoE NSSL (C,C1) in healthy mice brain showing an increase in the amount of actively targeted NSSL and their payload accumulated compared to passively targeted NSSL. We further compared the therapeutic efficacy of these targeted NSSL loaded with MPS in the acute EAE mice model (Fig 6B, Table 6). SJL mice were injected on days 10, 12, 14 post immunization with saline (control) or 10mg/kg NSSL-MPS (with or without the targeting peptides attached to their surface). Gaillard et al. [57] tried to enhance brain penetration of NSSL-MPS by using a targeted NSSL-MPS formulation (based on our previously published remote loaded NSSL-MPS formulation conjugated to the brain-targeting ligand glutathione), demonstrating a slight advantage (at best) in the MBP-EAE rats over the passively targeted formulation, apparent only 24 h after treatment. In the present study, although at the end of the experiment, a week after the third and last injection, MPS levels were significantly higher in the targeted liposomes-treated groups compared to the NSSL-MPS-treated group (Fig 6B). Both targeted formulations (ApoE- and β-amyloid-conjugated NSSL) did not show any therapeutic advantages over the passively targeted NSSL formulation, although all three formulations significantly ameliorated disease severity compared to control group. Furthermore, β-amyloid NSSL-MPS treatment was significantly less efficacious than passively targeted NSSL-MPS treatment. Treatment with ApoE NSSL-MPS showed similar therapeutic efficacy to treatment with NSSL-MPS (mean burden of disease: 0.9±0.1 and 1±0.1, respectively) compared to control (mean burden of disease 2.4±0.17). Treatment with β-amyloid NSSL-MPS lowered slightly the mean maximal clinical scores (3.2±0.31) and the mean burden of disease (1.7±0.14) compared to the control group (3.9±0.35, 2.4±0.17). Therefore, the improved penetration into the brain did not correlate with improved efficacy. The actively targeted NSSL probably distribute less uniformly throughout the diseased brains while remaining attached to cells close to the blood vessels, similarly to what was demonstrated before for tumor targeted NSSL [58]. In addition, BBB disruption in EAE mice, which enables penetration of passively targeted NSSL, might explain the lack of therapeutic advantage of actively targeted NSSL which may be able to extravasate the intact BBB over the passively targeted NSSL.

Bottom Line: For the NSSL-MPS we also compared the effect of passive targeting alone and of active targeting based on short peptide fragments of ApoE or of β-amyloid.Our results clearly show that for NSSL-MPS, active targeting is not superior to passive targeting.The highly efficacious anti-inflammatory therapeutic feature of these two nano-drugs meets the criteria of disease-modifying drugs and supports further development and evaluation of these nano-drugs as potential therapeutic agents for diseases with an inflammatory component.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Membrane and Liposome Research, Department of Biochemistry and Molecular Biology, Institute for Medical Research Israel-Canada (IMRIC), The Hebrew University-Hadassah Medical School, Jerusalem, Israel.

ABSTRACT
The present study shows the advantages of liposome-based nano-drugs as a novel strategy of delivering active pharmaceutical ingredients for treatment of neurodegenerative diseases that involve neuroinflammation. We used the most common animal model for multiple sclerosis (MS), mice experimental autoimmune encephalomyelitis (EAE). The main challenges to overcome are the drugs' unfavorable pharmacokinetics and biodistribution, which result in inadequate therapeutic efficacy and in drug toxicity (due to high and repeated dosage). We designed two different liposomal nano-drugs, i.e., nano sterically stabilized liposomes (NSSL), remote loaded with: (a) a "water-soluble" amphipathic weak acid glucocorticosteroid prodrug, methylprednisolone hemisuccinate (MPS) or (b) the amphipathic weak base nitroxide, Tempamine (TMN). For the NSSL-MPS we also compared the effect of passive targeting alone and of active targeting based on short peptide fragments of ApoE or of β-amyloid. Our results clearly show that for NSSL-MPS, active targeting is not superior to passive targeting. For the NSSL-MPS and the NSSL-TMN it was demonstrated that these nano-drugs ameliorate the clinical signs and the pathology of EAE. We have further investigated the MPS nano-drug's therapeutic efficacy and its mechanism of action in both the acute and the adoptive transfer EAE models, as well as optimizing the perfomance of the TMN nano-drug. The highly efficacious anti-inflammatory therapeutic feature of these two nano-drugs meets the criteria of disease-modifying drugs and supports further development and evaluation of these nano-drugs as potential therapeutic agents for diseases with an inflammatory component.

No MeSH data available.


Related in: MedlinePlus