Upload
free-escort-service
View
222
Download
0
Embed Size (px)
Citation preview
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
1/55
PROTEIN AND PEPTIDE DRUG
DELIVERY SYSTEMS Guided by :
Mrs. M.R.P. RAOPharmaceutics department)
Presented by :Mr.Dhanesh H. Sali.
AISSMS COLLEGE OF PHARMACY, PUNE
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
2/55
CONTENTS
IntroductionProtein and peptide drugsParenteral drug delivery systems
Non Parenteral drug delivery systemsDevelopment of drug delivery systemsInsulinConclusionReferences
2
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
3/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
4/55
4
Scientific advances in molecular and cell biology
have resulted in the development of two newbiotechnologies. The first utilizes recombinant DNAto produce protein products. The second technology is hybridoma technology.
Various proteins and peptides drugs are epidermalgrowth factor, tissue plasminogen activator.
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
5/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
6/55
MARKETED PROTEINS IN FREEZE DRIEDFORMULATIONS
Product Formulation Route Indication
Metrodin FSH 75 IU i.m. Induction ofovulation
Pergonal FSH and LH i.m. infertility
Profasi HCG i.m. Infertility
Elspar Asparginase i.m. i.v. Leukemia
Glucagon Glucagon i.m. i.v. s.c. Hypoglycemia
Acthar Corticotropin i.m. i.v. s.c. HormoneDeficiency 6
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
7/55
MARKETED PEPTIDES IN READY TO USEFORMULATIONSProduct Formulation Route Indication
Pitressin 8-ArginineVasopressin
i.m. s.c. Post operativeabdominaldistension
Lupron Leuprolide s.c. Prostatic cancer
Syntocinon Oxytocin i.m. i.v. Labour induction
Sandostatin Octreotide s.c. Intestinal tumour
Calcimar Salmon calcitonin s.c. hypercalcemia 7
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
8/55
SUSTAINED RELEASE DOSAGE FORMS
Product Formulation Route Indication
Lupron Leuprolide i.m. Prostaticcancer
H.P.Acthargel
ACTH i.m. s.c. Antidiuretic
Pitrressintannate inoil
Vasopressin tannate
i.m. Endocrinecancer
8
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
9/55
P ROTEIN AND PEPTIDE DRUGS
They are therapeutically effective only by parenteralroute.Repeated injections are required.
Therapeutic applications of these drugs rely onsuccessful development of viable delivery systemsto improve their stability and bioavailability.
9
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
10/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
11/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
12/55
PLGA biodegrades into lactic and glycolicacids. These acids enter into TCA cycle andthen eliminated as carbon dioxide andwater. Injectable controlled release
formulations of certain drugs are formulatedusing lactide/glycolide copolymers. Suchdrugs are LHRH, calcitonin, insulin.Nanoparticles made of PLGA, albuminpolystyrene have potential for targeted drugdelivery.
12
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
13/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
14/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
15/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
16/55
BASIC PRINCIPLES OF SELFREGULATED DEVICES
COMPETITIVE DESORPTION :
Insulin molecules with covalently attached sugarmolecules are used that is also known asglycosylated insulin. These are complementary to major binding site ofconA. It is carbohydrate binding protein.
Glycosylated insulin can be bound to conA andreversibly displaced from conA by glucose in directproportion of glucose concentration.
16
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
17/55
MEMBRANE CONTROLLEDDEVICES
Glucose oxidase is immobilized in cross linkedpolymers. In absence of external glucose aminegroups are unprotonated and membrane porosity issuch that insulin molecules are unable to diffuse
out.Glucose when diffused into membranes getsoxidized by glucose oxidase to gluconic acid whichprotonates amino groups due to which membraneporosity increases and now insulin can diffuse out.
17
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
18/55
EROSION CONTROLLEDDEVICES
Reaction between glucose and glucose oxidasegenerates gluconic acid. Poly(ortho esters)polymers erodes as pH decreases. Release ofinsulin can be modulated using this approach.
18
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
19/55
19
Non parenteral systemic delivery
These routes are useful for long term therapy. Without permeation enhancers lower bioavailability
is achieved when these routes are used. Lower bioavailability is due to poor mucosal
permeability. Sodium tauroglycocholate is commonly used
penetration enhancer.
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
20/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
21/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
22/55
Transdermal route :Skin has very low proteolytic activity.
Two types of iontophoresis are used :DIRECT CURRENT MODEPULSE CURRENT MODE
Vaginal route :Especially useful to deliver hormones.Not much accepted in developing countries.
22
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
23/55
DEVELOPMENT OF DELIVERY SYSTEMS FOR PEPTIDE AND PROTEIN BASED PHARMACEUTICALS
Considerations are to be given for following aspects:
Preformulation and Formulation considerationsPharmacokinetic considerations Analytical considerationsRegulatory considerations
23
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
24/55
P REFORMULATION AND F ORMULATION CONSIDERATIONS
Preformulation data is to be generated for followingaspects :
Isoelectric pointpH solubility profilepH stability profileExcipient compatibilities
24
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
25/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
26/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
27/55
P HARMACOKINETIC CONSIDERATIONS
Basal insulin secretion in healthy subjects showscircadian rhythm with peak time at 15:00 hrs. It has been suggested that larger amount of insulinis needed in afternoon and night.Hence delivery systems could be designed byconsidering such aspects.
27
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
28/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
29/55
U.V. SPECTROSCOPY Proteins containing aromatic amino acid residuessuch as phenyl alanine, tyrosine, tryptophan can bedetected by u.v. spectroscopy. Ultraviolet spectroscopy can be used for in processquality control. Protein aggregates scatter u.v. light andabsorbance increases. Hence u.v. spectroscopycan be used to monitor protein aggregation.
29
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
30/55
30
BRADFORD ASSAY :This assay employs the principle that in the presence of
proteins absorption maximum of coomassie brilliant bluedye changes from 465nm to 595nm.
BIURET TEST :
Structure of biuret and proteins are similar. Biuret inpresence of proteins or peptides reduces copper tocuprous ions in alkaline solutions and colour complex isdeveloped.
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
31/55
THERMAL ANALYSISDifferential scanning calorimetry (DSC) is gainingwidespread use as a tool for investigatingtransitions of confirmation as a function oftemperature and, more importantly, the effect ofpotential stabilizing excipients in a protein solution.The apex of the endothermic peak is the transitiontemperature between native and partially unfoldedconfirmations.
31
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
32/55
ELECTROPHORESISMost often used technique for protein products issodium dodecyl sulphate polyacrylamide gelelectrophoresis (SDS-PAGE).Proteins are denatured by boiling in the SDS
solution. All charges of protein are masked bynegative charge of dodecyl sulphate.Thus protein moves on polyacrylamide gel strictlyon basis of size of protein molecule. This technique is useful for determining molecularweight of proteins.For visualization of proteins on the gel reagentsused are silver nitrate, coomassie brilliant blue dye.
32
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
33/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
34/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
35/55
PROTEIN INSTABILITIES
The degradation of proteins and peptides can bedivided into two main categories :
1. Those that involve a covalent bond.2. Those involving a conformational change. This
process is often referred to as denaturation.
35
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
36/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
37/55
37
OXIDATIONSulphur containing amino acids are prone to oxidation.
MAILLARD REACTIONIn the maillard reaction the carbonyl group (RCH=O)
from glucose can react with the free amino group in apepide to form a Schiff base. This reaction is acidcatalysed.
DIMERISATION AND POLYMERIZATIONInsulin forms a small amount (about 1%) of covalentdimer and polymer during two years cold storage.Production of these species increases as temperatureincreases.
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
38/55
DENATURATION
o Specific confirmation is required for proteins to exertpharmacological and physiological activities.Denaturation is a process of altering protein confirmation.Heat, organic solvents, high salt concentration,
lyophilization can denature proteins.Protein confirmation refers to the specific tertiarystructure, which is determined by the primary andsecondary structures and the disulfide bonds and is held
together by three forces : hydrogen bonding, salt bridges,and hydrophobic interactions.
38
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
39/55
COMMON STABILIZERS
SERUM ALBUMIN :It can withstand heating to 60 o C for 10 hours. At pH 2 albumin molecule expands and elongates butcan return to native confirmation reversibly. Also, it shows
good solubility.Mechanism for such behaviour may be one of thefollowing :
Inhibition of surface adsorption or cryoprotection.
39
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
40/55
40
AMINO ACIDS
Glycine is most commonly used stabilizer.Mechanism of action of amino acids as stabilizers maybe one of the following :Reduce surface adsorption.
Inhibit aggregate formation.Stabilize proteins against heat denaturation.
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
41/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
42/55
POLYHYDRIC ALCOHOLS ANDCARBOHYDRATES :
They contain CHOH-CHOH- groups which areresponsible for stabilizing proteins. They stabilizeproteins against denaturation caused by elevatedtemperature or by freeze drying or by freeze thaw
cycles.Many important therapeutic proteins and peptidesare derived from blood such as immune globulin,coagulation factors. For viral destruction
pasteurization at 60o
C for 10 hours is needed.Hence thermal stability is needed. Long chainpolyhydric alcohols are more effective asstabilizers. e.g. sorbitol, xylitol. 42
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
43/55
Mechanism of action as stabilizers for polyhydric alcoholsis that they have effect on structure of surrounding watermolecules which strengthens hydrophobic interactions inprotein molecules. Mechanism of action as stabilizers for carbohydrates is
that they provide dry network that provides significantsupport for protection.Polyhydric alcohols used are sorbitol, mannitol, glycerol,PEG.
Carbohydrates used are glucose, mannose, sucrose,ribose.
43
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
44/55
44
ANTI-OXIDANTSThiol compounds such as thioacetic acid, triethanolamine,
reduced glutathione and metal chelants such as EDTA areused as antioxidants.
MISCELLANEOUS
Certain enzymes can be stabilized by using compoundshaving similar structures of enzymes. e.g. Glucose stabilizesglucoamylase while aspargine stabilizes asparginase.Compounds forming stable complex through ionic interactionwith proteins can stabilize proteins.Calcium is essential for thermal stability of certain amylases orproteases.
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
45/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
46/55
46
Protamine zinc insulin (PZI) contain an excess quantity ofzinc to retard absorption. According to BRITISH
PHARMACOPOEIA of 1958, a phosphate buffer is addedto each individual vial containing acidified solution ofinsulin, protamine and zinc so that pH is between 6.9 to7.3. The preparation is compounded in final container by
mixing PZI and buffer in filling operations.Main problem in using insulin is to avoid plasmaconcentration fluctuations. To solve this problem variousformulations are available varying in onset and durationof action.
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
47/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
48/55
USP INSULIN TYPE STRENGTHS
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
49/55
USP INSULIN TYPE STRENGTHS
Insulin injection (regular insulin) U-40 mixed,U-100 mixed, U-500
Isophane insulin suspension (NPHInsulin)
U-40 mixed,U-400 mixed
Isophane insulin suspension(70%) and insulin injection (30%)
U-100
Insulin zinc suspension (Lenteinsulin)
U-40 mixed,U-100 mixed
Extended insulin zinc suspension(Ultra lente insulin)
U-40 mixed, U-100 mixed
Prompt insulin zinc suspension(semilente insulin)
U-40 mixed, U-100 mixed
Protamine zinc insulin suspension(PZI Insulin)
U-40 mixed, U-100 mixed 49
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
50/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
51/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
52/55
REFERENCES
1) Agrawal S, Udupa N, Protein and peptide drug delivery :recent advances. In : Jain NK, editor. Progress in controlledand novel drug delivery systems. 1 st ed. Delhi : CBSPublishers; 2004.p.184-204.
2) Chien YW : Novel drug delivery systems. 2 nd ed. New York :Marcel Dekker Inc; 2005.p.631-745.
3) Yu Chang John Wang : Parenteral products of proteins andpeptides. In : Lieberman HA, Avis KE, editors. Pharmaceuticaldosage forms : Parenteral medications, volume 1. 2 nd ed. NewYork Marcel Dekker Inc; 2005.p.283-320.
4) Block JH, Beale JM. Wilson and Gisvold s textbook of organicmedicinal and pharmaceutical chemistry. 11 th ed. Philadelphia: Lippincott Williams and wilkins; 2005.p.851-852.
52
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
53/55
53
5) Patel NK, Pharmaceutical suspensios. In :Lachman l, Lieberman HA, Kanig JL, editor. Thetheory and practice of pharmacy. 3 rd ed. Mumbai :Varghese Publishing House; 1987.P.488-489.
6) Aulton ME : Pharmaceutics : The science ofdosage form design. 2 nd ed. Toronto : Churchilllivingstone; 2006.p.544-553.
7) Poon CY : Clinical Analysis. In : Troy DB, editor.Remington : The Science of Dosage form Design.21 st ed. Volume 1. Philadelphia : Lippincott
Williams and wilkins; 22005.p.577-578.8) www.ida.lib (accessed on 15/4/2010.)9) Rang HP, Dale MM : Pharmacology. 5 th ed. Toronto
: Churchill livingstone; 2003.p.386-388.
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
54/55
8/11/2019 32129428 Protein and Peptide Drug Delivery Systems
55/55
55