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Presentation byPresentation by
SUNILBOREDDYSUNILBOREDDY
(M. Pharmacy)(M. Pharmacy)
Resealed ErythrocyteResealed Erythrocyte
As Drug CarrierAs Drug Carrier
Content
Introduction
Erythrocytes as drug carrier
Basic concept of erythrocytes
Advantages & limitation
Source & Isolation of Erythrocytes
Method of drug Loading
Characterization
Drug release, Storage
Application
Conclusion
INTRODUCTION
Current Research: Aimed at development of drug delivery
system to get max. therapeutic effect.
Idea of drug carrier system:to get maximum
therapeutic effects, Elimination of undesirable side effects and Increase
patient compliance.
Magic Bullet Concept: Paul Ehrlich proposed. Decrease
toxic effect and increase therapeutic effect to the targeted cell.
Resealed Erythrocytes
Properties as carrier :-
Appropriate size, shape to permit passage through the
capillaries.
Biocompatible & minimum toxic side effects
Minimum leakage before target site is achieved
Should be able to carry broad spectrum of drugs
Appreciable stability during storage period
Should have sufficient space & should carry adequate amounts
of drugs
Erythrocytes
Erythro= red
Cytes = cell
Biconcave discs, anucleate.
Filled with hemoglobin
(Hb), a protein that
functions in gas transport
Contain the plasma
membrane protein spectrin
and other proteins that:
Give erythrocytes their
flexibility
Allow them to change shape as
necessary
Erythrocytes
CHEMICAL CONSTITUENTS:
Water(63%)
Lipid(0.5%)
Glucose(0.8%)
Minerals(0.7%)
Non-hemoglobin protiens(0.9%)
Methehemoglobin(0.5%) and
Hemoglobin (33.67%)
Erythrocytes
Healthy adult male= 4.5 millions/µml
Healthy adult female= 4.8 millions/µml
Matured RBCs have no nucleus Ribosome &
Mitochondria . Therefore all space is available for
drug carrier.
Reticulocytes- immature erythrocytes
Advantages of erythrocyte as carrier:-
Biodegradable
Isolation is easy
Non immunogenic
large volume of drug can be encapsulated in small volume of erythrocytes
Prolong systemic activity of drug
Protection from premature degration
Prodrug concept (Bioreactor)
ReduceAdverse Effect
Peptide & Enzyme Delivery
Disadvantages :-
Possibility of Leakage of cells & dose dumping
Molecule Alter Physiology Of cell
Source, fractionation & isolation of
erythrocytes
Source:- mice, cattle, pig, dog, sheep, goat, monkey, chicken, rat,
rabbit & human
Whole blood can be collected by venipuncture or from orbital sinus in
heparinized tube
Red blood cells can be harvested & centrifuged(2500rpm/5min at 40
c .
Different centrifugal force & different buffer composition(PBS,pH7.4)
for different species is used
Fresh blood is used for loading of drugs.
Methods Of Drug Loading InMethods Of Drug Loading In
ErythrocytesErythrocytes
1. Hypo-osmotic lyses method
a) Dilution method
b) Dialysis method
c) Pre-swell method
d) Isotonic osmotic lyses method
2. Electric breakdown method
3. Endocytosis method
4. Membrane perturbation method
5. Normal transport method
6. Lipid fusion method
Dilutional Haemolysis
RBC Membrane ruptured RBC Loaded RBC
Resealed Loaded RBC
0.4% NaCl
Hypotonic
Drug
Loading buffer
Resealing
buffer
Incubation at
250
c
Efficiency  1-8%
Enzymes
delivery
Hypotonic med
Isotonic med.
Washed
Isotonic Osmotic Lysis
RBC Isotonically ruptured
RBC
Chemical – urea, polyethylene, polypropylene, and NH4Cl
Physical rupturing
Chemical
rupturing
Drug Isotonic
Buffer
Loaded RBC
Resealed RBC
Incubation at
250
C
Preswell Dilutional Haemolysis
RBC
0.6%w/v NaCl Swelled
RBC Drug + Loading
buffer
5 min incubation
at 0 0
c
Loaded
RBC
Incubation at 25
0
c
Resealing Buffer
Resealed
RBC
Efficiency  72%
Fig:- Preswell Method
Dialysis
RBC
Phosphate
buffer
+
80 %
Haematocri
t value
Placed in dialysis
bag with air bubble
Dialysis bag placed in 200ml of lysis
buffer with mechanical rotator 2hrs. 4c.
DrugLoading
buffer
Loaded RBCDialysis bag placed in Resealing buffer
with mechanical rotator 30 min 37c.
Resealed RBC
Efficiency  30-45%
Entrapment by Endocytosis
RBC
Drug
Suspension
+
Buffer containing ATP,
MgCl2, and CaCl2
At 250
C
Loaded RBC Resealing Buffer
Resealed RBC
Fig;- Entrapment By Endocytos Method
Membrane perturbation method
RBC
Amphotericin B
e.g. Chemical agents
Increased
permeability of RBC
Resealing Buffer
Drug
Resealed RBC
In vitro characterizationIn vitro characterization
In vitro characterization
Mechanism of release of resealed erythrocytes
Shelf & storage stability of resealed erythrocytes
In vivo survival & immunological consequences
Some Important Aspects Of Resealed
Erythrocytes
Applications of resealed erythrocytes
Erythrocytes as carrier for enzymes
Erythrocytes as carrier for drugs
Erythrocytes for drug targeting
Drug targeting to reticuloendothelial system
Drug targeting to liver
-Treatment of liver tumors
-Treatment of parasitic diseases
-Removal of RES iron overload
-Removal of toxic agents
Applications of resealed erythrocytes
Delivery of antiviral agents
Oxygen deficiency therapy
Microinjection of macromolecules
Novel systems
Nanoerythrosomes
Erythrosomes
References
Jain.S., Jain.N.K., resealed erythrocytes as drug carriers,
Edited Jain N.K., Controlled And Novel Drug Delivery,
New Delhi, CBS publishers, New Delhi, 2004, 256-281.
Vyas S.P., Khar R.K., Targeted And Controlled Drug
Delivery: Novel Carrier Systems, New Delhi, CBS
publisher, 2004, 387-413.
Resealed erythrocytes

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Resealed erythrocytes

  • 1. Presentation byPresentation by SUNILBOREDDYSUNILBOREDDY (M. Pharmacy)(M. Pharmacy) Resealed ErythrocyteResealed Erythrocyte As Drug CarrierAs Drug Carrier
  • 2. Content Introduction Erythrocytes as drug carrier Basic concept of erythrocytes Advantages & limitation Source & Isolation of Erythrocytes Method of drug Loading Characterization Drug release, Storage Application Conclusion
  • 3. INTRODUCTION Current Research: Aimed at development of drug delivery system to get max. therapeutic effect. Idea of drug carrier system:to get maximum therapeutic effects, Elimination of undesirable side effects and Increase patient compliance. Magic Bullet Concept: Paul Ehrlich proposed. Decrease toxic effect and increase therapeutic effect to the targeted cell.
  • 4. Resealed Erythrocytes Properties as carrier :- Appropriate size, shape to permit passage through the capillaries. Biocompatible & minimum toxic side effects Minimum leakage before target site is achieved Should be able to carry broad spectrum of drugs Appreciable stability during storage period Should have sufficient space & should carry adequate amounts of drugs
  • 5. Erythrocytes Erythro= red Cytes = cell Biconcave discs, anucleate. Filled with hemoglobin (Hb), a protein that functions in gas transport Contain the plasma membrane protein spectrin and other proteins that: Give erythrocytes their flexibility Allow them to change shape as necessary
  • 7. Erythrocytes Healthy adult male= 4.5 millions/µml Healthy adult female= 4.8 millions/µml Matured RBCs have no nucleus Ribosome & Mitochondria . Therefore all space is available for drug carrier. Reticulocytes- immature erythrocytes
  • 8. Advantages of erythrocyte as carrier:- Biodegradable Isolation is easy Non immunogenic large volume of drug can be encapsulated in small volume of erythrocytes Prolong systemic activity of drug Protection from premature degration Prodrug concept (Bioreactor) ReduceAdverse Effect Peptide & Enzyme Delivery Disadvantages :- Possibility of Leakage of cells & dose dumping Molecule Alter Physiology Of cell
  • 9. Source, fractionation & isolation of erythrocytes Source:- mice, cattle, pig, dog, sheep, goat, monkey, chicken, rat, rabbit & human Whole blood can be collected by venipuncture or from orbital sinus in heparinized tube Red blood cells can be harvested & centrifuged(2500rpm/5min at 40 c . Different centrifugal force & different buffer composition(PBS,pH7.4) for different species is used Fresh blood is used for loading of drugs.
  • 10. Methods Of Drug Loading InMethods Of Drug Loading In ErythrocytesErythrocytes 1. Hypo-osmotic lyses method a) Dilution method b) Dialysis method c) Pre-swell method d) Isotonic osmotic lyses method 2. Electric breakdown method 3. Endocytosis method 4. Membrane perturbation method 5. Normal transport method 6. Lipid fusion method
  • 11. Dilutional Haemolysis RBC Membrane ruptured RBC Loaded RBC Resealed Loaded RBC 0.4% NaCl Hypotonic Drug Loading buffer Resealing buffer Incubation at 250 c Efficiency  1-8% Enzymes delivery Hypotonic med Isotonic med. Washed
  • 12. Isotonic Osmotic Lysis RBC Isotonically ruptured RBC Chemical – urea, polyethylene, polypropylene, and NH4Cl Physical rupturing Chemical rupturing Drug Isotonic Buffer Loaded RBC Resealed RBC Incubation at 250 C
  • 13. Preswell Dilutional Haemolysis RBC 0.6%w/v NaCl Swelled RBC Drug + Loading buffer 5 min incubation at 0 0 c Loaded RBC Incubation at 25 0 c Resealing Buffer Resealed RBC Efficiency  72% Fig:- Preswell Method
  • 14. Dialysis RBC Phosphate buffer + 80 % Haematocri t value Placed in dialysis bag with air bubble Dialysis bag placed in 200ml of lysis buffer with mechanical rotator 2hrs. 4c. DrugLoading buffer Loaded RBCDialysis bag placed in Resealing buffer with mechanical rotator 30 min 37c. Resealed RBC Efficiency  30-45%
  • 15. Entrapment by Endocytosis RBC Drug Suspension + Buffer containing ATP, MgCl2, and CaCl2 At 250 C Loaded RBC Resealing Buffer Resealed RBC Fig;- Entrapment By Endocytos Method
  • 16. Membrane perturbation method RBC Amphotericin B e.g. Chemical agents Increased permeability of RBC Resealing Buffer Drug Resealed RBC
  • 17. In vitro characterizationIn vitro characterization
  • 19. Mechanism of release of resealed erythrocytes Shelf & storage stability of resealed erythrocytes In vivo survival & immunological consequences Some Important Aspects Of Resealed Erythrocytes
  • 20. Applications of resealed erythrocytes Erythrocytes as carrier for enzymes Erythrocytes as carrier for drugs Erythrocytes for drug targeting Drug targeting to reticuloendothelial system Drug targeting to liver -Treatment of liver tumors -Treatment of parasitic diseases -Removal of RES iron overload -Removal of toxic agents
  • 21. Applications of resealed erythrocytes Delivery of antiviral agents Oxygen deficiency therapy Microinjection of macromolecules Novel systems Nanoerythrosomes Erythrosomes
  • 22. References Jain.S., Jain.N.K., resealed erythrocytes as drug carriers, Edited Jain N.K., Controlled And Novel Drug Delivery, New Delhi, CBS publishers, New Delhi, 2004, 256-281. Vyas S.P., Khar R.K., Targeted And Controlled Drug Delivery: Novel Carrier Systems, New Delhi, CBS publisher, 2004, 387-413.