Peptides and Proteins Formulation

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Peptides and Proteins Formulation

• Mahmoud R. Jaafari, PhD
• Prof. of Pharmaceutics

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Introduction
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Overview

• Structure of peptide and protein
• Solubility of peptides and proteins
• The stability of peptides and proteins
• Formulation of therapeutic peptides and proteins
  as parenteral solutions
• Formulation of therapeutic peptides and proteins
  as lyophilized powder
• Formulation of therapeutic peptides and proteins
  as parenteral suspensions

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Protein structure
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Protein Folding

• Highly simplified schematic representation of the
  folding of a polypeptide chain in water, In the
  unfolded chain, side-chain and main-chain groups
  interact primarily with water, even if they are
  hydrophobic and the interaction is unfavorable.
  Burying the hydrophobic groups in the interior of
  a compact structure enables them to interact with
  each other (blue line), which is favorable, and
  leaves polar side chains on the surface where
  they can interact with water (red lines). The
  polar backbone groups that are buried along with
  the hydrophobic side chains must make hydrogen
  bonds to each other, as bulk water is no longer
  available.

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Bonds that stabilize folded proteins
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Solubility of peptides and proteins

• 1) Nature of peptides and proteins
  (Hydrophobicity of peptides and proteins)

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Solubility of peptides and proteins

• 2) Effects of salt concentration

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Solubility of peptides and proteins

• 3) Effects of pH
• Solubility of beta-lactoglobulin as a function of
  pH at several NaCl concentrations

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Solubility of peptides and proteins

• 4) Effects of organic solvents
• 5) Effects of temperature

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The stability of Peptides and Proteins
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Physical Instability

• 1) Denaturation
• Heat Reversible and irreversible
• The optical rotation at 366 nm as a function of
  temperature of bovine pancreatic ribonuclease A

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Physical Instability
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Physical Instability Denaturation

• Changes of pH
• Detergents
• Water soluble organic compound
• Salts Hofmeister series

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Physical Instability aggregation

• Aggregation vs self-association

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Physical Instability

• Precipitation

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Physical Instability

• Surface adsorption

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Chemical Instability Deamidation

• pHlt4

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Chemical Instability Deamidation

• pHgt6

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Chemical Instability

• Prevention of deamidation

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Chemical Instability Protein Oxidation
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Chemical Instability

• Prevention of Protein Oxidation

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Chemical Instability Racemisation

• Alkaline Media

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Chemical Instability Proteolysis
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Chemical Instability Beta elimination
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Chemical Instability Disulphid formation
(Exchange)
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Formulation of peptides and proteins as
parenteral solutions

• Introduction
• Preformulation studies
• Formulation development

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Formulation of peptides and proteins as
parenteral solutions

• Buffer system

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Formulation of peptides and proteins as
parenteral solutions

• pH of vehicle
• Solubility of peptides and proteins

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Selection of solvent system (Cosolvents)
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Antimicrobial preservation
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Choice of container
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Pharmaceutical excipients in the formulation of
peptides and proteins

• Albumin
• Amino acids
• Carbohydrates

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Pharmaceutical excipients in the formulation of
peptides and proteins

• Chelating agents and antioxidants

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Pharmaceutical excipients in the formulation of
peptides and proteins

• Cyclodextrins

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Pharmaceutical excipients in the formulation of
peptides and proteins

• Surfactants

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Pharmaceutical excipients in the formulation of
peptides and proteins

• Polyhydric alcohols
• Polyethylene glycol

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Selection of excipients DSC
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Selection of excipients DSC
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Accelerated stability studies

• Gel Filtration Column Chromatography

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Accelerated stability studies GPC
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Accelerated stability studies GPC
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Formulation of peptides and proteins as
lyophilized dosage form

• Introduction
• Process Details

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Formulation of peptides and proteins as
lyophilized dosage form
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Formulation of peptides and proteins as
lyophilized dosage form

• Protein denaturation during lyophilization
• Freeze- Thaw stability
• Stability during drying

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Formulation components for lyophilization

• Buffers
• Bulking agents
• Tonicity modifiers
• Stabilizer

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Formulation components for lyophilization

• Cryoprotectives

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Formulation components for lyophilization

• Lyoprotectives

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Formulation of peptides and proteins as
parenteral suspensions

• Introduction

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Types of suspensions and particle formation

• In situ particle formation
• Crystalline suspensions
• Amorphous suspensions
• Crystalline and amorphous mixture
• Crystalline and solution mixture
• Combination of particle and vehicle

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In situ particle formation Crystalline
suspensions
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In situ particle formation Crystalline
suspensions
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In situ particle formation Crystalline
suspensions