spektroskopia epr w badaniu nanoleków
DESCRIPTION
Spektroskopia EPR w badaniu nanoleków. Ryszard Krzyminiewski 1,2 , Bernadeta Dobosz 1,2 , Joanna Kurczewska 3 , Grzegorz Schroeder 3 , Magdalena Hałupka-Bryl 2 , Magdalena Bednarowicz 2 , Tomasz Kubiak 1 1 Zakład Fizyki Medycznej, Wydział Fizyki UAM, Umultowska 85, 61-614 Poznań, - PowerPoint PPT PresentationTRANSCRIPT
Spektroskopia EPR w badaniu nanoleków
Ryszard Krzyminiewski1,2, Bernadeta Dobosz1,2, Joanna Kurczewska3, Grzegorz Schroeder3, Magdalena Hałupka-Bryl2,
Magdalena Bednarowicz2, Tomasz Kubiak1
1Zakład Fizyki Medycznej, Wydział Fizyki UAM, Umultowska 85, 61-614 Poznań, 2Centrum NanoBioMedyczne, Umultowska 85, 61-614 Poznań,
3Zakład Chemii Supramolekularnej, Wydział Chemii UAM, Umultowska 89B, 61-614 Poznań,
Electron Paramagnetic Resonance (EPR) Spectroscopy
Warunek rezonansowy
hν=gμBB
EPR – the nuclear hyperfine interaction
- Oddziaływania nadsubtelne: oddziaływanie spinowego momentu magnetycznego elektronu ze spinowym momentem magnetycznym jądra.
- Wpływ na parametry widma mają oddziaływania dipolowe i wymienne
Znacznik spinowy
Czas korelacji τ informuje o dynamice
molekuł w danym środowisku, ich
rotacji itd
ΔH(0) szerokość linii centralnej a I(+1), I(0) i
I(-1) to amplitudy linii niskopolowej,
centralnej i wysokopolowej
EPR – correlation time and the spectrum of the radical NO.
10-12 s
10-11 s
10-10 s
10-8 s
10-9 s
EPR X-band
L-band
Materiały
Rheumatoid arthritis (RA):-chronic, systemic inflammatory disorder, affecting the joints, causing pain and stiffness, leading to destruction of the articular cartilage.-characterized by development of a pannus tissue-free radicals/reactive oxygen species (ROS) play an important role in RA inflammation.
CHRONIC INFLAMMATION
ROS scavenging activity plays an importnant role
in suppresing inflammation caused by
oxidative stressArthritis
Reactive Oxygen Species
Reactive Oxygen Species (ROS): chemically reactive molecules containing oxygen causing oxidative demage.
oxid
ative
str
ess
higher risk of systemic diseases e.g. Alzheimer, cancer, atherosclerosis
and…
…they play important role in chronic inflammation
pathogenesis
Materiały
ROS Scavenger
Antioxidant for excess generation of ROS
TEMPO( 2,2,6,6-tetramethylpiperidine-1-oxyl)
Low molecular ROS scavengers are easily eliminated from the system resulting in poor therapeutic effect.
N
O
Polyion Complex (PIC) Redox Flower Micelle
TEMPO(ROS scavanger)
polyion core(nanoreservoir
for drugs)
PMNT-PEG-PMNT triblock copolymer
hydrophobic hydrophilic hydrophobic
HS
O OS
H
HN
NO
HN
NO
m n m
N N N NN N N N
MATERIAL
PEG: poly(ethylene glycol)
PMNT: poly[p-4-(2,2,6,6-tetramethyl-1-piperidine-1-oxyl)aminomethylstyrene
injectable fluid which exhibis in situ gel formation
N N N NN N N N N N N N
N N N N
Injctable PIC hydrogel
protonated
pH
unprotonated
pH
+ + + + + + + +
the novel redox flower micelle developed by utilizing the pH-responsive character of PMNT-PEG-PMNT triblock c
opolymer
COO- COO-COO-COO-
electrostatic interactions
protonatedpH
unprotonatedpH
+
+
+
+
+
+
+
+ + +
+
+
++
+
_
_
_
__
_ _self- assembley disassembley
and gelation in 37 ℃ and 150mM ionic strenght
flower micelle solution PIC hydrogel
eeasy asy to to operaoperatete
bbiocompatibiliocompatibil
pprolongrolonginging duration of ROS scaven duration of ROS scavengersgers
llocal controllocal controlling ofing of drug release drug release
Advantages:0
2
4
6
8
10
12
14
0 50 100 150 200
micelle 1 day
micelle over 30 days
hydrophilic PEG shell(prevents aggregation, biocompatible)
chloroform
DMF
H2O
dry polymer TEMPO crystal
micelles
150K 280K
Linear transformation methodThe shape of the signal can be described by:
- a function describing the shape and width of particular spectral lines, e.g. Gauss
- a function describing positions and intensities of particular components of the signal
The Fourier Transform of F(x)
Determination of the function (x)
F x K x x x dx( ) ( ) ( )
K x x( )
( )x
dxiyxxFyF exp)(2)(~ 2/1
dyixyyKyF
x )exp()(~)(~
)2()( 2/1
Fig. PMNT-PEG-PMNT in H2O, DMF and chloroform (5mg/mL) peak-to-peak line width (ΔHpp) versus temperature [K]
g – Factor Hp [mT] DMF 2,0054 333,9water 2,0055 332,7chloroform 2,0058 333,0
Materiały
Materiały
Scheme 1. Schematic representation of the magnetic nanocarrier studied
Materiały
Schemat syntezy nanocząstek magnetytu pokrytych chitozanem i znakowanych TEMPO z dołaczonym antybiotykiem
Materiały
Mikrofotografie TEM A,20nm, B50nm, C10nm magnetycznych nanocząstek
Widma EPR znakowanych TEMPO nanocząstek magnetytu z dopaminą
Fig. 4. Changes of resonance field (Hr) versus temperature for investigated sample with (FC) and without field cooling ( ZFC).
Fig. 6. Changes of g-factor value versus temperature for investigated sample with (FC) and without field cooling (ZFC).
Zmiany parametrów widm EPR w funkcji temperatury i tzw. FC i ZFC
Widma EPR magnetytu pokrytego chitozanem i znakowanego TEMPO , temperatura 120K rejestracja ZFC i FC , badania orientacji w polu magnetycznym
Centralny fragment widma EPR 254K i 132K.
Zależności temperatorowe nanocząstek magnetytu parametrów EPR:szerokości linii, intensywności i wsp. g *linia szeroka od rdzenia
Fig. 5. The temperature dependences of fast rotation correlation times of nitroxidespin probe for the orientations 0 and 90.
Czasy korelacji
Tomografia EPR
Wolne rodnikiWolne rodniki
Wolne rodnikiWolne rodniki
Wolne rodnikiWolne rodniki
Wolne rodnikiWolne rodniki
Dyfuzja w ciągu 3 godzinDyfuzja w ciągu 3 godzin
Współczynniki dyfuzji nanocząstek w hydrożelu
Dziękuję za uwagę
Nie mam żadnych wykresów Hałupki
Materiały
AVAILABLE TREATMENTS
first-line drugs to reduce inflammation
and pain:NSAIDs
GC
second-line drugs to prevent
joints destruction:
DMARDs
Dexamethasone - DEX, a member of the GC class of steroid inflammatory and immunosuppressing drug
Prednisolone - PRD, a synthetic glucocorticoid, a derivative of cortisol, which is used to treat a variety of inflammatory diseases- active metabolite of the drug prednisone
LONG TERM TREATMENT CAUSING MANY SIDE EFFECTS
Current strategies are to design nanosystem for delivery of
therapeutic agents specifically to the site of inflammation, therefore
avoiding potential systemic and off-target unwanted effects.
HSO O
SHn
HS-PEG-SH
SO O
Sn
CH=CH2
Cl
・ S-PEG-S ・
SO O
Sn
HH
ClCl
m m
PCMS-PEG-PCMS
NH2N O
(4-amino-TEMPO)
PMNT-PEG-PMNT Triblock Copolymer
. .
PCMS-PEG-PCMS
-radical telomerization of CMS using SH-PEG-SH as telogen – 60 ℃, 24h in oil bath-purification by reprecipitation by 30-fold amount of diethyl ether-freeze-drying with benzene-amination of PCMS-PEG-PCMS with 4-amino TEMPO in DMSO -dialysis against methanol (48h)-evaporation-freeze-drying
AIBNtoluene
PMNT-PEG-PMNT synthesis
Fig. 7. Changes of correlation time (τ) versus temperature for investigated sample with (FC) and without field cooling (ZFC).