Recombinant Protein Therapeutics Flashcards
What makes disulphide bonds in insulin’s structure significant and how do they influence its function?
Insulins two peptide chains (A+B) are linked by disulphide bonds, essential for maintaining its biologically active structure. The correct formation of these bonds is critical for its ability to bind to insulin receptors and regulate blood levels
What are the primary challenges associated with producing recombinant protein like insulin in microbial systems like E. coli?
producing insulin in such systems requires overcoming issues such as incorrect folding, inclusion body formation, and post-translational modifications, which E.coli cannot naturally perform.
What is the MoA of GLP-1 analogues and their role in type 2 diabetes compared to insulin analogues?
GLP-1 analogues enhance insulin secretion, inhibit glucagon release, slow gastric emptying, and reduce appetite, these don’t require injections. Insulin analogues regulate glucose uptake
GLP-1 analogue example?
semaglutide, exenatide
insulin analogue example?
insulin lispro, aspart, glulisine
how does hexamer-monomer conversion of insulin influence the PK of different insulin formations?
hexamer form of insulin is more stable but slower to absorb, while monomers diffuse more rapidly into the bloodstream. This conversion is crucial in the design of insulin formations, where fast-acting insulins require rapid dissociation into monomers
do hexamers dissociate fast or slow?
slow
do monomers dissociate fast or slow?
fast into the bloodstream
why is insulin glargine formulated at pH 4, and how does this formulation influence its absorption profile?
to ensure solubility. Upon injection, the change in pH causes it to precipitate, leading to slow, sustained release into the bloodstream
what is the role of albumin binding in the prolonged action of insulin detemir and what is its implication therapeutically?
it binds to serum albumin due to its fatty acid modification, allowing for slow dissociation and extended duration of action. This prolonged action is advantageous in reducing the frequency of insulin injections.
how does the concept of PK/PD profiling apply to the development of long-acting insulin analogues?
PK/PD profiling helps in designing long-acting insulin analogues by analysing how the drug is absorbed, distributed, and excreted, and how these processes affect the duration and intensity of its therapeutic effect
what are the key quality control measures employed when manufacturing biotherapeutics like insulin?
monitoring for correct protein folding, purity, potency, and sterility, as well as ensuring the post-translational modifications are accurate
what insulin analogue is engineered by reversing the AA at positions 28 and 29 of the B chain?
lispro
what is the advantage of insulin hexamers over monomers in drug formulation?
hexamers are more stable for storage, though monomers are more reactive and faster-acting once dissociated in the bloodstream
what is recombinant protein therapy, and why is it significant in modern medicine?
it involves the use of proteins produced through recombinant DNA technology to treat chronic diseases. It allows for mass production of proteins like insulin, essential for treating diseases like diabetes
how is insulin produced using recombinant DNA technology?
by inserting the gene for human insulin into bacterial or yeast cells. These cells then express the insulin protein, which is harvested and purified for therapeutic use
what is an example of a short-acting insulin analogue?
Lispro - act quickly after injection
what is an example of a long-acting insulin analogue?
Glargine - provide stable release of insulin over a longer period
what happens when insulin is injected?
the stable storage hexamer form dissociates into monomers, which are active and can regulate blood sugar levels
how has insulin been engineered to create short-acting insulin analogues like insulin Lispro?
by reversing the order of the amino acids proline and lysine at positions 28 and 29 of the B-chain. This modification allows the insulin to dissociate from hexamers more quickly after injection, leading to faster absorption.
how does Oramed’s Protein Oral Delivery (POD) technology advance the possibility of oral insulin delivery?
it protects proteins like insulin from enzymatic degradation in the GIT and enhances their absorption across the intestinal wall. This could make oral insulin delivery a reality, replacing injections.
