Stimuli-responsive polysaccharides
based NPs for controlled and
sustained release of antineoplastic
drugs
Antonio Di Marti...
What are Stimuli Responsive Polymers (SRP)?
 High performance materials
 Show a sharp change in properties upon a small ...
SRP : Importance in drug delivery
Blood 7.35-7.45 Stomach 1.0-3.0 Colon 7.0-7.5
6.0-6.5
5.0-6.0
4.5 -5.0
 pH in different...
Aims of the work
• Preparation and characterization of polysaccharides
based NPs for therapeutic and diagnostic applicatio...
Methods
Materials : Antineoplastic drugs
Doxorubicin (DOX) Temozolomide (TMZ) 5-Fluorouracil (5-FU)
 Anti-metabolites
 Head, nec...
Materials : Carriers
Chitosan (CS)
Polygalacturonic
acid (PgA)
Alginic acid (Alg) Carboxy modified
Iron (Fe-COOH)
CS-Alg
C...
Methods : CS-Alg and CS-PgA NPs
Polyanion solution
Step I: Add the solution containing drug to polyanion solution
Drug(s) ...
Methods : Chitosan – Fe NPs
 Stirring
 Ultrasound
 Temperature
O. A. Guselnikova, M. V.Gromov, A. I. Galanov, Adv. Mat....
Methods : Characterization
 Dynamic Light Scattering
 Scanning Electron Microscopy
 Transmission Electron Microscope
 ...
Method : Encapsulation and Release
 Encapsulation Efficiency
 preparation media (pH 5.5)
 physiological media (pH 7.4)
...
Results : NPs characterization
CS-Alg
 d : 110 nm
 z-pot. + 35 mV
CS-PgA
 d : 130 nm
 z-pot : + 33 mV
CS-Fe
 d : 135 ...
Results : CS-Fe NPs - MRI
 T1 relaxation
High fat content tissues : Bright
Water filled tissues : Dark
.
 T2 relaxation
...
Results : Encapsulation
CS-Alg
pH 5.5 61 % 57% 45%
pH 7.4 26% 24% 31%
CS-PgA CS-Fe
pH 5.5 52 % 64% 43%
pH 7.4 31% 37% 28%
...
Results : Co-Encapsulation
pH 5.5 50% 55% 46%
pH 7.4 20% 21% 18%
pH 5.5 58% 57% 59%
pH 7.4 27% 24% 35%
pH 5.5 61 % 54% 49%...
Results : Release Kinetic – CS-Alg
0
20
40
60
80
100
0 20 40 60 80 100 120
Cumulativerelease(%)
Time (h)
0
20
40
60
80
100...
Results : Release Kinetic – CS-PgA
0
20
40
60
80
100
0 20 40 60 80 100 120
Cumulativerelease(%)
Time (h)
0
20
40
60
80
100...
 Chitosan based nanoparticles for therapeutic and diagnostic application
has been prepared
 NPs present size less than 1...
Short term perspective
 Cell uptakes studies
 Efficacy In vitro in different tumoral cell lines
 In Vivo MRI
Results : Co-Encapsulation
TMZ + 5-FU
mg/mg
TMZ + DOX
mg/mg
5-FU + DOX
mg/mg
CS-Alg (w/w = 2) TMZ
280
5-FU
220
TMZ
310
DOX...
Method : Encapsulation Efficiency
 Effect of pH
 UV-Vis
 266 nm 5-FU
 325 nm TMZ
 480 nm DOX
100(%) 




 
...
Polysaccharide based nanoparticles for encapsualtion and release of antineoplastic drugs
Polysaccharide based nanoparticles for encapsualtion and release of antineoplastic drugs
Polysaccharide based nanoparticles for encapsualtion and release of antineoplastic drugs
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Polysaccharide based nanoparticles for encapsualtion and release of antineoplastic drugs

nanoparticles, chitosan, iron nanoparticles, doxorubicin, temozolomide , 5-fluorouracil, controlled release
Published on: Mar 4, 2016
Published in: Science      
Source: www.slideshare.net


Transcripts - Polysaccharide based nanoparticles for encapsualtion and release of antineoplastic drugs

  • 1. Stimuli-responsive polysaccharides based NPs for controlled and sustained release of antineoplastic drugs Antonio Di Martino
  • 2. What are Stimuli Responsive Polymers (SRP)?  High performance materials  Show a sharp change in properties upon a small or modest modification in environmental condition  Diverse range of applications  Optical systems  Coating  Diagnostic  Tissue engineering  Drug delivery
  • 3. SRP : Importance in drug delivery Blood 7.35-7.45 Stomach 1.0-3.0 Colon 7.0-7.5 6.0-6.5 5.0-6.0 4.5 -5.0  pH in different tissue and cellular compartment Tumor cells extracellular matrix pH is lower than in healthy cells
  • 4. Aims of the work • Preparation and characterization of polysaccharides based NPs for therapeutic and diagnostic application • Encapsulation and co-encapsulation of three anticancer drugs • Effect of pH on the release kinetic
  • 5. Methods
  • 6. Materials : Antineoplastic drugs Doxorubicin (DOX) Temozolomide (TMZ) 5-Fluorouracil (5-FU)  Anti-metabolites  Head, neck, colon, skin  Easy to handle  Good stability  Side effects  Alkylating agent  Short plasma t ½  Grade IV glioma  Metastic melanoma  Alkylating agent  High therapeutic efficacy  Widely used  Side effects  Encapsulation in nanoparticles structure to prolong and maintain the drug concentration in the therapeutic window  Reduction of side effects
  • 7. Materials : Carriers Chitosan (CS) Polygalacturonic acid (PgA) Alginic acid (Alg) Carboxy modified Iron (Fe-COOH) CS-Alg CS-Fe CS-PgA Therapeutic Therapeutic + Diagnostic
  • 8. Methods : CS-Alg and CS-PgA NPs Polyanion solution Step I: Add the solution containing drug to polyanion solution Drug(s) solution Drug(s) + Polyanion in solution Step II: Add drop-wise A to B under magnetic stirring Drug(s) + Polyanion in solution (A) CS solution (B) Nanoparticles in suspension
  • 9. Methods : Chitosan – Fe NPs  Stirring  Ultrasound  Temperature O. A. Guselnikova, M. V.Gromov, A. I. Galanov, Adv. Mat. Res. 2014, 1040, 309-313. Step I: -COOH modified Fe NPs preparation Step II: coating and loading CS Drug
  • 10. Methods : Characterization  Dynamic Light Scattering  Scanning Electron Microscopy  Transmission Electron Microscope  Magnetic properties  Morphology  Size  z-potentiaL  Stability  Magnetic Resonance Imaging
  • 11. Method : Encapsulation and Release  Encapsulation Efficiency  preparation media (pH 5.5)  physiological media (pH 7.4)  Human Serum (HS)  Simulated Gastric Fluid (SGF) : pH 2  Preparation media (PM) : pH 5.5  Physiological solution (PS) : pH 7.4  Release kinetic ( T = 37 ºC)  UV-Vis  Absorbance at different wavelength
  • 12. Results : NPs characterization CS-Alg  d : 110 nm  z-pot. + 35 mV CS-PgA  d : 130 nm  z-pot : + 33 mV CS-Fe  d : 135 nm  z-pot : + 30 mV  After 1 month size increase of only 20 %  Great shelf-life  No particular storage conditions required  uncoated  coated  d : 10 nm  z-pot : - 25 mV Coating influence Magnetic response
  • 13. Results : CS-Fe NPs - MRI  T1 relaxation High fat content tissues : Bright Water filled tissues : Dark .  T2 relaxation Water filled tissues : Bright High fat content tissue : Dark MRI Signal suppression
  • 14. Results : Encapsulation CS-Alg pH 5.5 61 % 57% 45% pH 7.4 26% 24% 31% CS-PgA CS-Fe pH 5.5 52 % 64% 43% pH 7.4 31% 37% 28% pH 5.5 49 % 55% 31% pH 7.4 23% 15% 12% DOX TMZ 5-FU More than 50 % of encapsulation = around 400 mg/mg
  • 15. Results : Co-Encapsulation pH 5.5 50% 55% 46% pH 7.4 20% 21% 18% pH 5.5 58% 57% 59% pH 7.4 27% 24% 35% pH 5.5 61 % 54% 49% pH 7.4 26% 21% 31% TMZ 5-FU TMZ DOX DOX 5-FU + + + CS-Alg CS-PgA CS-Fe Drugs are well balanced in the system !!!
  • 16. Results : Release Kinetic – CS-Alg 0 20 40 60 80 100 0 20 40 60 80 100 120 Cumulativerelease(%) Time (h) 0 20 40 60 80 100 0 20 40 60 80 100 120 Cumulativerelease(%) Time (h) 0 20 40 60 80 100 0 20 40 60 80 100 120 Cumulativerelease(%) Time (h) HS pH7.4 pH5.5 pH2 HS pH7.4 pH5.5 pH2 HS pH7.4 pH5.5 pH2  Temperature : 37 º C  Increase pH release speed up  Reduction of Initial burst effect
  • 17. Results : Release Kinetic – CS-PgA 0 20 40 60 80 100 0 20 40 60 80 100 120 Cumulativerelease(%) Time (h) 0 20 40 60 80 100 0 20 40 60 80 100 120 Cumulativerelease(%) Time (h) 0 20 40 60 80 100 0 20 40 60 80 100 120 Cumulativerelease(%) Time (h) HS pH7.4 pH5.5 pH2 HS pH7.4 pH5.5 pH2 HS pH7.4 pH5.5 pH2  Temperature : 37 º C  Increase pH release speed up  Reduction of Initial burst effect
  • 18.  Chitosan based nanoparticles for therapeutic and diagnostic application has been prepared  NPs present size less than 150 nm and high stability up to one month  CS-Fe NPs show high T1 and T2 suppression  High encapsulation efficiency ( > 50 %) for single or multiple loading  Release rate can be increase or decrease according with pH  Reduction of initial burst effect compared with other systems
  • 19. Short term perspective  Cell uptakes studies  Efficacy In vitro in different tumoral cell lines  In Vivo MRI
  • 20. Results : Co-Encapsulation TMZ + 5-FU mg/mg TMZ + DOX mg/mg 5-FU + DOX mg/mg CS-Alg (w/w = 2) TMZ 280 5-FU 220 TMZ 310 DOX 270 5-FU 270 DOX 420 CS-PgA (w/w = 2) TMZ 210 5-FU 340 TMZ 260 DOX 190 5-FU 280 DOX 350 CS-Fe (w/w = 5) TMZ 180 5-FU 270 TMZ 120 DOX 190 5-FU 130 DOX 210 pH : 5.5 pH : 7.4 TMZ + 5-FU mg/mg TMZ + DOX mg/mg 5-FU + DOX mg/mg CS-Alg (w/w = 2) TMZ 280 5-FU 220 TMZ 290 DOX 310 5-FU 270 DOX 420 CS-PgA (w/w = 2) TMZ 210 5-FU 340 TMZ 260 DOX 190 5-FU 280 DOX 350 CS-Fe (w/w = 5) TMZ 90 5-FU 110 TMZ 190 DOX 200 5-FU 120 DOX 310  Average value SD up to 10%
  • 21. Method : Encapsulation Efficiency  Effect of pH  UV-Vis  266 nm 5-FU  325 nm TMZ  480 nm DOX 100(%)         t ft D DD EE  preparation media (pH 5.5)  physiological media (pH 7.4)  EE = Encapsulation Efficiency  Dt = total amount of drug (mg/mL)  Df = amount of free drug after encapsulation (mg/mL)

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