recombinant dna technology in medicine...recombinant dna technology in medicine 50 years of...
TRANSCRIPT
![Page 1: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/1.jpg)
RECOMBINANT DNA TECHNOLOGY IN MEDICINE50 years of recombinant drugs
Asist. Dr. Matjaž Ravnikar, M Pharm
Chair of Pharmaceutical Biology
Faculty of Pharmacy, Ljubljana
![Page 2: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/2.jpg)
RECOMBINANT DNA TECHNOLOGY
BIOTECHNOLOGY
RECOMBINANT DNA
TECHNOLOGY
![Page 3: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/3.jpg)
RECOMBINANT DNA TECHNOLOGY
Recombinant DNA (rDNA) technology refers to the process of joining DNA molecules from two different sources and inserting them into a host organism, to generate products for human use.
![Page 4: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/4.jpg)
APPLICATIONS OF RECOMBINANT DNA TECHNOLOGY
Therapeutic products
Vaccines
Recombinant proteins
Vectors
Growth hormones
Anticancer drugs
Genetically modified products
GM vegetables
GM crops
GM microbes
GM animals
Diagnosis
Gene therapy
CRISPR
Monitoring devices
Therapeutic strategies
Energy applications
Biohydrogen
Bioethanol
Biomethanol
Biobutanol
Recombinant DNA Technology
![Page 5: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/5.jpg)
SMALL MOLECULES AND PROTEINS
Discovered by chance Very similar to proteins in the body
![Page 6: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/6.jpg)
DIABETES (Type 1)
![Page 7: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/7.jpg)
History of Diabetes treatment
-Until 1920s deadly disease with no cure
-Porcine and bovine insulin (allergic reactions)
-2 tones of pig pancreatic tissue for 250 ml of Insulin
-Recombinant (human) insulin (1980s)
![Page 8: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/8.jpg)
Advencements in Insulin therapy
![Page 9: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/9.jpg)
From Gene to Protein
![Page 10: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/10.jpg)
Polymerase chain reaction
![Page 11: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/11.jpg)
Polymerase chain reaction
-Kary Mullis 1983
-thermo-stabile polymerase
https://www.youtube.com/watch?v=iQsu3Kz9NYo
![Page 12: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/12.jpg)
Principles of drug discovery
Receptors & ligands!
![Page 13: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/13.jpg)
Cell communication & phisiological responce
![Page 14: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/14.jpg)
Agonists and antagonists
![Page 15: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/15.jpg)
PROCESS OF RECOMBINANT DRUG PRODUCTION
![Page 16: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/16.jpg)
Searching for new drug candidates
![Page 17: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/17.jpg)
Peptide candidates screening
- Phage Display
- Bacteriophages
- Libraries of peptides
- Up to 1012 combinations
- Sequencing of positive clones
![Page 18: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/18.jpg)
Development of recombinant cell line
![Page 19: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/19.jpg)
Expression and purification of recombinantprotein
![Page 20: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/20.jpg)
RECOMBINANT MOLECULES IN CLINICAL USE
-Enzymes and regulatory proteins
(Human growth hormone, Insulin,…)
-Antibodies
(Cancer therapy)
-Vaccines
(HPV virus)
-Proteins for diagnostics
(affinity molecules)
-Gene therapy
(nucleic acids, recombinant vectors)
![Page 21: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/21.jpg)
Pros and Cons of recombinant proteins
Pros:
-high efficiency
-homology to endogenous proteins in our body
-few side effects
-new therapy options
Cons:
-expensive to produce
-protein structure usually requires parenteral application
-extensive purification, sterility required
-systemic effect
![Page 22: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/22.jpg)
ANTIBODIES
![Page 23: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/23.jpg)
Types of monoclonal antibodies
![Page 24: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/24.jpg)
Antibodies targeting cancer cells
![Page 25: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/25.jpg)
Varieties of antibody fragments
![Page 26: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/26.jpg)
Recombinant vaccines
![Page 27: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/27.jpg)
DNA vaccines
![Page 28: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/28.jpg)
FUSION PROTEINS
-Chemical fusion
-Recombinant fusion
![Page 29: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/29.jpg)
Fusion proteins advantages
- Easier cleaning (affinity tags)
- Directed expression (signaling peptide, extracellular)
- Visualization (fluorescent peptides)
- Enhancing cathalytic activity
- Enhancing stability
- Enhancing solubility
- Improving expression levels
- …
![Page 30: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/30.jpg)
Example: design and preparation ofrecombinant probiotic
![Page 31: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/31.jpg)
GENE THERAPY(fixing genetic problem at its source)
Cancer therapies
![Page 32: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/32.jpg)
In-vivo and Ex-vivo gene transfer
![Page 33: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/33.jpg)
First trials
-Adenosine deaminase deficiency leads to immunodeficiency
("Bubble kids")
Danger of developing leukemia (insertional mutagenesys)!
![Page 34: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/34.jpg)
Drawbacks of early gene therapies
-can damage vital genes
-can trigger lethal immune response to viral carrier
However…
-In therapy: Zolgensma® (1.8MEur) - Spinal muscular atrophy (AAV)
![Page 35: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/35.jpg)
"Zinc finger" nucleases
Artificial nucleases, capable of recognizing specific DNA sequences
![Page 36: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/36.jpg)
TALENs (Transcription Activator-Like Effector Nucleases)
-tandem repeats of 34 amino acids, containing 2 variable AA
![Page 38: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/38.jpg)
Future of recombinant DNA technology in medicine and ethical concerns
- Reproductive cloning: mice, cattle, pigs, goats, rabbits, and cats
- From repairing to enhancing human performance?
![Page 39: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/39.jpg)
Thank you for your attention!
![Page 40: RECOMBINANT DNA TECHNOLOGY IN MEDICINE...RECOMBINANT DNA TECHNOLOGY IN MEDICINE 50 years of recombinant drugs Asist. Dr. Matjaž Ravnikar, M Pharm Chair of Pharmaceutical Biology Faculty](https://reader034.vdocuments.us/reader034/viewer/2022042502/5f764568d3d59b2e48621d12/html5/thumbnails/40.jpg)