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Pharmaceutical Research

, Volume 34, Issue 1, pp 58–72 | Cite as

Matrix Metalloproteinase Responsive Delivery of Myostatin Inhibitors

  • Alexandra C. Braun
  • Marcus Gutmann
  • Regina Ebert
  • Franz Jakob
  • Henning Gieseler
  • Tessa Lühmann
  • Lorenz Meinel
Research Paper

Abstract

Purpose

The inhibition of myostatin - a member of the transforming growth factor (TGF–β) family - drives regeneration of functional skeletal muscle tissue. We developed a bioresponsive drug delivery system (DDS) linking release of a myostatin inhibitor (MI) to inflammatory flares of myositis to provide self-regulated MI concentration gradients within tissues of need.

Methods

A protease cleavable linker (PCL) – responding to MMP upregulation – is attached to the MI and site-specifically immobilized on microparticle surfaces.

Results

The PCL disintegrated in a matrix metalloproteinase (MMP) 1, 8, and particularly MMP-9 concentration dependent manner, with MMP-9 being an effective surrogate biomarker correlating with the activity of myositis. The bioactivity of particle-surface bound as well as released MI was confirmed by luciferase suppression in stably transfected HEK293 cells responding to myostatin induced SMAD phosphorylation.

Conclusions

We developed a MMP-responsive DDS for MI delivery responding to inflammatory flare of a diseased muscle matching the kinetics of MMP-9 upregulation, with MMP-9 kinetics matching (patho-) physiological myostatin levels.

Graphical Abstract

Schematic illustration of the matrix metalloproteinase responsive delivery system responding to inflammatory flares of muscle disease. The protease cleavable linker readily disintegrates upon entry into the diseased tissue, therby releasing the mystatin inhibitor.

KEY WORDS

bioorthogonal chemistry bioresponsive drug delivery controlled release myostatin protease cleavable linker 

ABBREVIATIONS

ActRIIB

Activin receptor IIB

au

Arbitrary units

BCA

Bicinchoninic acid

BSA

Bovine serum albumin

CuAAC

Copper(I)-catalyzed azide-alkyne cycloaddition

DBCO

Dibenzocyclooctyne

DDS

Drug delivery system

DM

Differentiation medium

DMD

Duchenne muscular dystrophy

ECM

Extracellular matrix

EDC

1-Ethyl-3-(3dimethylaminopropyl)carbodiimide

Fmoc

N-α-(9-Fluorenylmethyloxycarbonyl)

GDF-8

Growth differentiation factor 8

HPLC

High-performance liquid chromatography

IGF-I

Insulin-like growth factor I

IL-1β

Interleukin 1β

MALDI-MS

Matrix-assisted laser desorption ionization mass spectrometry

MI

Myostatin inhibitor

MMP

Matrix metalloproteinase

MSTN

Myostatin

MyHC

Myosin heavy chain

NF-kB

Nuclear factor-kappa B

NHS

N-hydroxysuccinimide

PCL

Protease cleavable linker

PEG

Polyethylene glycol

PMMA

Poly(methyl methacrylate)

RLU

Relative light unit

RT-PCR

Real time polymerase chain reaction

SBE

SMAD binding element

SC

Satellite cell

SPAAC

Strain-promoted azide-alkyne cycloaddition

SPPS

Solid phase peptide synthesis

TG

Transglutaminase (human, fXIIIa)

TGF–β

Transforming growth factor beta

THPTA

Tris(3-hydroxypropyltriazolylmethyl)amine

TNF-α

Tumor necrosis factor α

Notes

ACKNOWLEDGMENTS AND DISCLOSURES

We thank Petra Knaus (Freie Universität Berlin, Germany) for providing us with the pGl3ti-SBE constructs and Melanie Krug for excellent technical assistance. We thank Dr. Joachim Nickel for providing C2C12 myoblast cells. The financial support of the Bavarian research foundation (grant # AZ-1044-12 ‘FORMOsA’) and the Deutsche Forschungsgemeinschaft (DFG; ME 3820/3-1) are gratefully acknowledged. H.G. is full time associate of Gilyos GmbH.

Supplementary material

11095_2016_2038_MOESM1_ESM.pdf (779 kb)
ESM 1 (PDF 779 kb)

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Institute for Pharmacy and Food ChemistryUniversity of WürzburgWürzburgGermany
  2. 2.Orthopedic Center for Musculoskeletal ResearchUniversity of WürzburgWürzburgGermany
  3. 3.GILYOS GmbHWürzburgGermany

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