Slide 1: WEL-COME Proteins and Peptides drug delivery system A Seminar on PROTEIN AND PEPTIDE DRUG DELIVERY SYSTEM : A Seminar on PROTEIN AND PEPTIDE DRUG DELIVERY SYSTEM By
Bhimrao K. Rupnoor
(M. Pharm., Sem. II)
Under the guidance of
Mr. K.K. Mali
Satara College of Pharmacy, Satara. CONTENTS : CONTENTS 3 Protein & Peptides
Structure of protein
Classification of protein
Parenteral delivery of protein and peptide
Ocular delivery of protein & peptide
References Protein & Peptide : Protein & Peptide 4 . Proteins: These are large organic compounds made of amino acids arranged in a linear chain and joined together by peptide bonds.
Protein > 50 amino acids
Peptides: These are short polymers formed from the linking, in a defined order, of α-amino acids.
peptide < 50 amino acids STRUCTURE OF PROTEIN : STRUCTURE OF PROTEIN 5 There are four types.
Primary structure- The amino acid sequence.
Secondary structure- Regularly repeating local structures stabilized by hydrogen bond.
Tertiary structure-Three dimensional structure of polypeptide
Quaternary structure-The structure formed by several protein molecules (polypeptide chains). Slide 6: 6 Protein Structure : Protein Structure 7 Lactate Dehydrogenase: Mixed α /β Immunoglobulin Fold: β Hemoglobin B Chain: α Classification of Proteins : Classification of Proteins According to their biological roles
Enzymes – Catalyses virtually all chemical reactions i.e. 6GDH
Transport proteins i.e. Haemoglobin of erythrocytes
Contractile or Motile proteins i.e. Actin and Myosin
Structural proteins i.e.Collagen in bones
Defense proteins i.e. Immunoglobulins and Antibodies
Regulatory proteins i.e. insulin
Nutrient and storage proteins i.e. Ovalbumin 8 FUNCTIONS : FUNCTIONS 9 Transport and storage of small molecules.
Coordinated motion via muscle contraction.
Mechanical support from fibrous protein.
Generation and transmission of nerve impulses.
Immune protection through antibodies.
Control of growth and differentiation via hormones. Slide 10: 10 Advantages of protein and
peptide drug delivery system Erythropoietin used for production of RBC.
Tissue plasminogen activator is used for Heart attack, Stroke.
Oxytocin maintain labor pain.
Bradykinin increases the peripheral circulation.
Somatostatin decrease bleeding in gastric ulcer.
Gonadotropin induce ovulation.
Insulin maintain blood sugar level. Disadvantages : Disadvantages Very large and unstable molecules.
Structure is held together by weak noncovalent forces.
Easily destroyed by relatively mild storage conditions and gastric juices.
Hard to obtain in large quantities. Problem with Proteins (in vivo – in the body) : Problem with Proteins (in vivo – in the body) Elimination by B and T cells
Proteolysis by endo/exo peptidases
Small proteins filtered out by the kidneys very quickly
Unwanted allergic reactions may develop (even toxicity)
Loss due to insolubility/adsorption 12 Slide 13: 13 Problem with Proteins (in vitro – in the bottle) : Problem with Proteins (in vitro – in the bottle) Noncovalent Covalent
Denaturation - Deamidation
Aggregation - Oxidation
Precipitation - Proteolysis
Adsorption 14 Covalent processes : Covalent processes 15 Deamidation - conversion of Asn-Gly sequences to a-Asp-Gly or b-Asp-Gly
Oxidation - conversion RSR’ to RSOR’, RSO2R’ or RSO3R’ (Met & Cys)
Proteolysis - Asp-Pro, Trypsin (at Lys) or Chymotrypsin (at Phe/Tyr) Slide 16: 16 How to Deal with These Problems Storage Formulation Delivery Storage : Storage Refrigeration
Freeze-Drying 17 Commonly used excipients For solving physical and chemical stability problems : Commonly used excipients For solving physical and chemical stability problems 24 Jan. 2010 18 Modern College of Pharmacy, Pune Slide 19: 19 Slide 20: 20 Protein Formulations Site Directed Mutagenesis : Site Directed Mutagenesis 21 Allows amino acid substitutions at specific sites in a protein
i.e. substituting a Met to a Leu will reduce likelihood of oxidation E343H PEGylation : PEGylation 22 PEG is a non-toxic, hydrophilic, FDA approved, uncharged polymer
Increases in vivo half life
Increases protease resistance
Increases stability PEGylation : PEGylation 23 + CH-CH-CH-CH-CH-CH-CH-CH-CH-CH
| | | | | | | | | |
OH OH OH OH OH OH OH OH OH OH Proteinylation : Proteinylation 24 Attachment of additional or secondary (nonimmunogenic) proteins for in vivo protection
Cross-linking with Serum Albumin
increases in vivo half life
Cross-linking or connecting by protein engineering with antibody fragments Proteinylation : Proteinylation 25 + Protein drug scfc (antibody) Formulation with permeabilizers : Formulation with permeabilizers 26 Salicylates (aspirin)
Metal chelators (EDTA) Slide 27: 27 Drug Delivery Parenteral Delivery of Proteins : Parenteral Delivery of Proteins 28 Intravenous
Intradermal Advantages and Disadvantages : Advantages and Disadvantages 29 Slide 30: 30 Polymeric Drug Delivery Polymers should be:
Natural- Chitosan, Dextrin
Synthetic- Polylactides/ glycolide Polyanhydrides
Polyphosphoesters Slide 31: 31 Release Mechanism Diffusion of drug out of the polymer Drug Release by Polymer Degradation Hydrolysis
Enzymatic (Phosphotases; Proteases etc.) Microsphere Drug delivery : Microsphere Drug delivery 32 Two types of microspheres Nonbiodegradable
e.g, ceramic particles
polyethylene co-vinyl acetate
polylactic-co-glycolic acid (PLGA) Magnetic Targeted Carriers (MTCs) : Magnetic Targeted Carriers (MTCs) 33 Founder of FeRx and pioneer of magnetic targeted drug delivery is Dr. Kenneth Widder Microparticles, composed of elemental iron and activated carbon
Drug is adsorbed into the MTCs and transported
The particles serve as delivery vehicles to the area of the tumor for site-specific targeting Liposomal Drug Delivery : Liposomal Drug Delivery 34 Spherical vesicles with a phospholipid bilayer E.g, Bleomycin encapsulated in thermo sensitive liposome enhanced antitumor activity and reduced normal tissue toxicity
Liposome have recently been used successfully as vehicles for vaccines Hydrogel Based Drug Delivery : Hydrogel Based Drug Delivery 35 Hydrogels are three dimensional networks of hydrophilic
polymers that are insoluble Hydrogel Based Drug Delivery : Hydrogel Based Drug Delivery 36 e.g Insulin Hydrogels can swell as a result of changes in pH, Temperature. Proteins in Pumps : Proteins in Pumps 37 Multiple potential interactions between the protein and the pump Infusaid Model 400 Implantable Pump Mechanical Insulin Pumps Emulsion and Cellular carriers : Emulsion and Cellular carriers Emulsion :
Emulsion can be used for parenteral delivery of protein and peptides. Multiple emulsion further prolong the release of drug.
e.g. subcutaneous administration of muramyl dipeptide in a w/o emulsion
Protein and peptides can be incorporated in erythrocytes to achieve the prolong release or targeting.
Resealed erythrocytes as delivery system for c-reactive protein, and mainly used to target liver and spleen. 38 Ocular Delivery of Peptide & Protein : Ocular Delivery of Peptide & Protein 39 Relevant anatomy and Physiology of the Human eye
Diameter of 23 mm
Outermost coat : the clear, transparent cornea and the white, opaque sclera
Middle layer : the iris anteriorly, the choroid posteriorly, and the ciliary body at the intermediate part
Inner layer : retina Peptides Useful in Ocular Pathology : Peptides Useful in Ocular Pathology 40 To treat infections and enzymes used to promote wound healing.
The drug is applied topically to the eye for a variety of conditions, including eyelid burns and corneal superficial punctate keratits .It is also used to treat optic neuritis.
Commercial bacitracin is a mixture of at least nine bacitracins, which is used for its antibacterial activity Slide 41: b. Chymotrypsin
Chymotrypsin is used clinically in the eye for enzymatic intracarpsular lens extraction.
Cyclosporine are a group of biologically active metabolites produced by Fungi. When cyclosporine A is administered by nonocular route in rats, it produces significant blood levels, but it can not be detected in tissues. 41 Advantages : Advantages 42 The mode of delivery is convenient, i.e., eye drops.
Systemic absorption is extremely rapid.
Avoid first-pass metabolism.
The formulation can be designed to prolong drug action and/or reduce drug concentrations to achieve consistent drug action with least side effects.
Drug delivery can be controlled precisely. Marketed formulations : Marketed formulations Slide 44: 44 : 45 Conclusion REFERENCES : REFERENCES 46 Frokjaer, Hovgard,Pharmaceutical formulation development of peptides and proteins,2000 .29-32,41-60.
Avis E. Kennith, Lieberman A, Herbert., Lachmam, Leon., Pharmaceutical dosage form: Parenteral Medication vol-1, Second edition, Mercel Dekker, USA, 284, 302, 312.
Robinson, R. Joseph. Controlled Drug Delivery : Fundamental &application, Second edition, Mercel Dekkar, USA, 7-14. Slide 47: 47 Mitra, K.Ashim, Opthalmic Drug delivery System, Second edition, Mercel Dekkar,464-73.
Jain N.K., Progress in Controlled & novel Drug delivery system. CBS publication; Delhi, 195.
Gupta H, Sharma A. Recent trends in protein and peptide drug delivery systems. Asian J Pharm 2009;3:69-75. Slide 48: 48 Thank you…