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The Evaluation of Therapeutic Efficacy and Safety Profile of Simvastatin Prodrug Micelles in a Closed Fracture Mouse Model

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ABSTRACT

Purpose

To evaluate the therapeutic efficiency of a micellar prodrug formulation of simvastatin (SIM/SIM-mPEG) and explore its safety in a closed femoral fracture mouse model.

Methods

The amphiphilic macromolecular prodrug of simvastatin (SIM-mPEG) was synthesized and formulated together with free simvastatin into micelles. It was also labeled with a near infrared dye for in vivo imaging purpose. A closed femoral fracture mouse model was established using a three-points bending device. The mice with established closed femoral fractures were treated with SIM/SIM-mPEG micelles, using free simvastatin and saline as controls. The therapeutic efficacy of the micelles was evaluated using a high-resolution micro-CT. Serum biochemistry and histology analyses were performed to explore the potential toxicity of the micelle formulation.

Results

Near Infrared Fluorescence (NIRF) imaging confirmed the passive targeting of SIM/SIM-mPEG micelles to the bone lesion of the mice with closed femoral fractures. The micelle was found to promote fracture healing with an excellent safety profile. In addition, the accelerated healing of the femoral fracture also helped to prevent disuse-associated ipsilateral tibia bone loss.

Conclusion

SIM/SIM-mPEG micelles were found to be an effective and safe treatment for closed femoral fracture repair in mice. The evidence obtained in this study suggests that it may have the potential to be translated into a novel therapy for clinical management of skeletal fractures and non-union.

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Abbreviations

ALP:

Alkaline phosphatase

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

BMD:

Bone mineral density

BS/TV:

Bone surface density

BUN:

Blood urea nitrogen

BV/TV:

Bone volume fraction

CON:

Vehicle-treated control group

Conn.D:

Connectivity density

CPK:

Creatine phosphokinase

DA:

Degree of anisotropy

Ecc:

Mean eccentricity

ELVIS:

Extravasation through leaky vasculature and inflammatory cell-mediated sequestration

FD:

Fractal dimension

FX:

Fracture

H&E:

Hematoxylin and eosin

IACUC:

Institutional animal care and use committee

micro-CT:

Micro-computed tomography

MMI:

Mean polar moment of inertia

NIRF:

Near infrared fluorescence

PEG:

Polyethylene glycol

ROI:

Region of interest

SIM:

Simvastatin

SIM/SIM-mPEG:

A micellar macromolecular prodrug formulation of simvastatin based on polyethylene glycol

SIMA:

Simvastatin acid

SMI:

Structure model index

Tb.N:

Trabecular number

Tb.Pf:

Trabecular bone pattern factor

Tb.Sp:

Trabecular separation

Tb.Th:

Trabecular thickness

TLC:

Thin layer chromatography

TP:

Total protein

VOI:

Volume of interest

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Acknowledgments & Disclosures

This study was supported in part by NIH (R01 AR062680) and Nebraska Arthritis Outcome Research Center (NAORC). We thank Mr. Arun Tatiparthi of Micro Photonics for his assistance with the micro-CT analysis. Y.Z. was supported by the Bukey Fellowship from University of Nebraska Medical Center.

Y.Z., Z.J., A.D., S.R.G. and D.W. are listed as co-inventors in a PCT patent application filed for the SIM/SIM-mPEG prodrug micelle technology.

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Corresponding author

Correspondence to Dong Wang.

Additional information

Yijia Zhang and Zhenshan Jia contributed equally to this work.

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Zhang, Y., Jia, Z., Yuan, H. et al. The Evaluation of Therapeutic Efficacy and Safety Profile of Simvastatin Prodrug Micelles in a Closed Fracture Mouse Model. Pharm Res 33, 1959–1971 (2016). https://doi.org/10.1007/s11095-016-1932-2

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  • DOI: https://doi.org/10.1007/s11095-016-1932-2

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