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Pharm (Industrial Pharmacy) Ist Resealed ErythrocyteAs Drug Carrier Content : 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:- : Introduction:- Current Research Idea of drug carrier system Magic Bullet Concept Resealed 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 Erythrocytes : 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 : 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 Slide 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) Reduce Adverse Effect Peptide & Enzyme Delivery Disadvantages :- Possibility of Leakage of cells & dose dumping Molecule Alter Physiology Of cell Source, fractionation & isolation of erythrocytes : 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 : 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 : Dilutional Haemolysis RBC Membrane ruptured RBC Loaded RBC Resealed Loaded RBC 0.4% NaCl Hypotonic Drug Loading buffer Resealing buffer Incubation at 250c Efficiency 1-8% Enzymes delivery Hypotonic med Isotonic med. Washed Isotonic Osmotic Lysis : 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 : Preswell Dilutional Haemolysis RBC 0.6%w/v NaCl Swelled RBC Drug + Loading buffer 5 min incubation at 0 0c Loaded RBC Incubation at 25 0c Resealing Buffer Resealed RBC Efficiency 72% Fig:- Preswell Method Dialysis : Dialysis RBC Phosphate buffer + 80 % Haematocrit value Placed in dialysis bag with air bubble Dialysis bag placed in 200ml of lysis buffer with mechanical rotator 2hrs. 4c. Drug Loading buffer Loaded RBC Dialysis bag placed in Resealing buffer with mechanical rotator 30 min 37c. Resealed RBC Efficiency 30-45% Electro-insertion or Electro-encapsulation : Electro-insertion or Electro-encapsulation RBC 2.2 Kv Current for 20 micro sec At 250 C Pulsation medium + + Drug Loading suspension 3.7 Kv Current for 20 micro sec Isotonic NaCl Loaded RBC Resealing Buffer Resealed RBC Fig:- Electro-encapsulation Method Entrapment By Endocytosis:- : 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 Slide 17: Membrane perturbation method RBC Amphotericin B e.g. Chemical agents Increased permeability of RBC Resealing Buffer Drug Resealed RBC In vitro characterization : In vitro characterization Slide 19: In vitro characterization Slide 20: 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 : 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 : Applications of resealed erythrocytes Delivery of antiviral agents Oxygen deficiency therapy Microinjection of macromolecules Novel systems Nanoerythrosomes Erythrosomes Conclusion : Conclusion References : 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. Indian Journal of Pharmaceutical Education & Research Vol. 43(4), Oct-Dec, 2009 , 375-386 Journal of Controlled Release 95 (2004) 27– 49 Any Questions or Suggestions? : Any Questions or Suggestions? Any Questions or Suggestions? Slide 26: THANK YOU ! You do not have the permission to view this presentation. In order to view it, please contact the author of the presentation.