RESEALED ERYTHROCYTE AS DRUG CARRIER

Presenting By: MAHESWARI.G Guided By: Prof. Mrs. A .PRAMEELA RANI GARU
M. Pharm , Ph.D

Content
Introduction

Erythrocytes as drug carrier

Basic concept of erythrocytes Advantages & limitation

Source & Isolation of Erythrocytes

Method of drug Loading Drug release, Storage

Application
Conclusion

Introduction: DEF: They are drug -loaded carrier erythrocytes

Idea of drug carrier system

Magic Bullet Concept by PAUL ERLICH

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
Healthy adult male= 4.5 millions/ml Healthy adult female= 4.8 millions/ml Matured RBCs have no nucleus Ribosome & Mitochondria . Therefore all space is available for drug carrier. Reticulocytes- immature erythrocytes

Resealed Erythrocytes
Properties as carrier :Appropriate size, shape.
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

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 Different centrifugal force & different buffer composition for different species is used Fresh blood is used for loading of drugs.

METHODS OF DRUG LOADING IN ERYTHROCYTES

Membrane perturbation

Electroencapsulation

Hypoosmotic lysis

Lipid fusion endocytosis

Dilution method

Preswell method

Osmotic lyses method

Dialysis method

Dilutional Haemolysis

0.4% NaCl

Drug Membrane ruptured RBC Loaded RBC

Loading buffer Incubation at 250c
Hypotonic med

RBC
Hypotonic

Resealing buffer

Resealed Loaded RBC

Enzymes delivery

Isotonic med

Washed

Isotonic Osmotic Lysis

Physical rupturing

RBC

Isotonically ruptured

RBC
Drug
Chemical rupturing

Isotonic Buffer

Loaded RBC Incubation at 250 C Resealed RBC

Chemical urea, polyethylene, polypropylene, and NH4Cl

Preswell Dilutional Haemolysis

RBC

0.6%w/v NaCl

Swelled

5 min incubation at 0 0c Drug + Loading buffer

Loaded

Incubation at 25 0c Resealing Buffer

Resealed

RBC

RBC

RBC

Fig:- Preswell Method

Dialysis

RBC
+
Phosphate buffer Loading buffer Placed in dialysis bag with air bubble Dialysis bag placed in 200ml of lysis buffer with mechanical rotator 2hrs. 4c.

Drug

Resealed

RBC

Dialysis bag placed in Resealing buffer with mechanical rotator 30 min 37c.

Loaded RBC

Electro-insertion or Electroencapsulation
RBC +
Pulsation medium At 250 C 2.2 Kv Current for 20 micro sec

Drug

3.7 Kv Current for 20 micro sec

Loaded RBC Isotonic NaCl Resealing Buffer Resealed

+
Loading suspension

RBC

Fig:- Electro-encapsulation Method

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

Amphotericin B

RBC
e.g. Chemical agents

Increased permeability of RBC

Drug Resealed Resealing Buffer

RBC

Advantages :Biodegradable & biocompatible 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 degradation Prodrug concept (Bioreactor) Reduce Adverse Effect Peptide & Enzyme Delivery

Disadvantages :Possibility of Leakage of cells & dose dumping Molecule Alter Physiology Of cell

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

CONCLUSION:The use of resealed erythrocytes looks promising for a safe and sure delivery of various drugs for passive and active targeting. However, the concept needs further optimization to become a routine drug delivery and much remains to be explored regarding the potential of resealed erythrocytes

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.
PHARMACEUTICAL TECHNOLOGY : A Review