sugar-modified ppi dendrimers as carriers of anti-leukemia drugs barbara klajnert-maculewicz...
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Sugar-modified PPI dendrimers as carriers of anti-leukemia drugs
Barbara Klajnert-Maculewicz
Department of General Biophysics
University of Lodz
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Linear polymer1930s
Branched polymer1960s
Dendrimer1980s
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dendrimer - gr. dendronarborol - lat. arborcascade molecule
D. Tomalia
F. Vögtle
G. Newcome
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dendrimer - gr. dendronarborol - lat. arborcascade molecule
dendrimer
cascade molecule
arborol
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• globular shape
• monodispersity
• big dimensions
• high reactivity
• good solubility
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DRUG DELIVERY
GENE CARRIERS ANTI-PRION ACTIVITY
ANTI-MICROBIAL ACTIVITY
ANTI-HIV ACTIVITY
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Dendrimers as drug carriers
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• prolonging the circulation time in blood
• protecting unstable drugs from the environment
• enhancing solubility of poorly soluble drugs
• achieving a controlled release and tissue targeting
• one molecule of dendrimer is capable to carry drugs at high density
• slower release than for a free drug
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Physiological nucleoside analogues (NAs)
Cytarabine is used to treat different forms of leukemia, including acute and chronic myelogenous (AML and CML) and acute lymphocytic leukemia (ALL).
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arabinose - cytosine
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Drug resistance mechanisms
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Conventional therapy with ara-C
Drug resistance
Dendrimers as carriers of ara-CTP
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Maltose-modified PPI dendrimer
PPI-OS-Mal G5 (approx. 30% maltose)
Dr. Dietmar AppelhansSynthesis: Fischer et al. Biomacromolec. 10 (2009) 1314-1325
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Advantages of sugar modification
non-hemolytical 1, 2
do not cause platelets aggregation 2
low toxicity in vitro 3
low toxicity in vivo 4
retained activity 1, 5, 6
1. B. Klajnert et al. Chem. Eur. J. 14 (2008) 7030-70412. B. Ziemba et al. J. Biomed. Mater. Res. A. 100 (2012) 28703. A. Janaszewska et al. New J. Chem. 36 (2012) 428-4374. B. Ziemba et al. J. Biomed. Mater. Res. A 99 (2011) 261-2685. M. Fischer et al. Biomacromolecules 11 (2010) 1314-13256. M. Ciołkowski et al. Colloid Surface B 95 (2012) 103-108
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+
• dendrimer and ara-CTP form complexes (ara-CTP/dendrimer=10)• complexes are stable• ara-CTP is protected against enzymatic degradation
ara-CTP
complexes:
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dephosphorylation
alkaline phosphatase
ATP adenosine
A. Szulc et al. New J. Chem. 36 (2012) 1610-1615
Protection agaist enzymatic degradation – FPLC method
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free ATPbound ATP
A. Szulc et al. New J. Chem. 36 (2012) 1610-1615
Protection agaist enzymatic degradation – FPLC method
A254 nm
retention time = 13 min
retention time = 19 min
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free ATPbound ATP
adenosine
retention time = 29 min
A. Szulc et al. New J. Chem. 36 (2012) 1610-1615
Protection agaist enzymatic degradation – FPLC method
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adenosine
retention time = 29 min
system PPI-m G5 + adenosine no complex!
A. Szulc et al. New J. Chem. 36 (2012) 1610-1615
Protection agaist enzymatic degradation – FPLC method
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no free ATPbound ATP
adenosineretention time = 29 minred line:
system PPI-m G5 + ATP +alkaline phosphatase
A. Szulc et al. New J. Chem. 36 (2012) 1610-1615
Protection agaist enzymatic degradation – FPLC method
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bound ATP
red line:system PPI-m G5 + ATP +alkaline phosphatase
26% of complexed ATPwas degraded.
A. Szulc et al. New J. Chem. 36 (2012) 1610-1615
Protection agaist enzymatic degradation – FPLC method
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1301
human acute lymphoblastic T cells
leukemia cell line: control cells:
PBMC
peripheral blood mononuclear cells
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Internalization of dendrimers into cells
fluorescein (FITC)
Flow cytometry
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Internalization of dendrimers into 1301 cells
fluorescein (FITC)
Confocal microscopy
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Cytotoxicity Alamar Blue assay
Cytotoxicity test with resazurin - in live cells it is reduced to resorufin:
exc=530 nmem=590 nm
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Dendrimer cytotoxicity
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Cytotoxicity - 1301 48h
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Cytotoxicity - 1301 48h
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Cytotoxicity - 1301 48h
*
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Cytotoxicity - 1301 72h
** * *
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Cytotoxicity - PBMC 48h
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Cytotoxicity - PBMC 72h
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Inhibitor of nucleoside transporters
NBMPR - nitrobenzylmercaptopurine
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1301
48 h 72 h
Inhibitors of NT in
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1301
Anti-leukemia activity:
> >
>> !
1.
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CONCLUSIONS:
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Acknowledgements
Dr. Dietmar AppelhansLeibniz-Institutefür PolymerforschungDresden e.V. Germany
Aleksandra Szulc, M.Sc.PhD studentUniversity of LodzPoland
TEAM project „Biological properties and biomedical applications of dendrimers”
BIODENDRIMER 2014
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Thank you.
BIODENDRIMER 2014
Faculty of Biology and Environmental Protection,Lodz, Poland