NUCLEIC ACID BASED DELIVERY SYSTEM.pptSx
- 1. NUCLEIC ACID BASED THERAPEUTIC DELIVERY SYSTEM:GENE THERAPY 1 MOLECULAR PHARMACEUTICS (NANOTECHNOLOGY AND TARGETED DRUG DELIVERY SYSTEMS) FACULTY-IN-CHARGE: Dr. PRASANTH MS ASSOCIATE PROFESSOR, DEPARTMENT OF PHARMACEUTICS PRESENTED BY: LANIYA NASRIN K 2nd SEMESTER M PHARM COLLEGE OF PHARMACEUTICAL SCIENCES, GOVERNMENT MEDICAL COLLEGE, THIRUVANANTHAPURAM
- 2. NUCLEIC ACID BASED THERAPEUTIC DELIVERY SYSTEM GENE THERAPY GERMLINE GENE THERAPY SOMATIC CELL GENE THERAPY 01 02 EX VIVO & IN VIVO GENE THERAPY 03 04 Table of contents 05 06 REFERENCES 2
- 3. 3 NUCLEIC ACID BASED THERAPEUTIC DELIVERY SYSTEM 01
- 4. 4 NUCLEIC ACID BASED THERAPEUTIC DELIVERY SYSTEM • A Nucleic acids is a complex, biochemical macromolecules which is composed of nucleotides chain that transfer genetic information. • They are the monomer components: a 5-carbon sugar, a phosphate group and a nitrogenous base • If the sugar is ribose, the polymer is RNA • If the sugar is deoxyribose, a variant of ribose, the polymer is DNA. • Deoxyribonucleic acid (DNA) • Ribonucleic acid (RNA) CLASSES OF NUCLEIC ACID
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- 6. 6 • Nucleic acid based therapeutic delivery system is the delivery of nucleic acid molecules into cells to alter physiological functions at genetic level, a powerful approach to treat a wide range of inherited and acquired disorders. • It is used for developing a most effective and safe delivery of gene in vivo • It become the most important objective for the medical prospects of gene therapy • It involves the transfer of the therapeutic agents to the targeted sites in the body for the treatment of several diseases
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- 8. 8 • The industries of pharmaceutical and biotechnology produces large variety of drugs based on nucleic acid • Many oligonucleotides and RNA interferences (RNAi) drugs are under clinical trial phase II • It includes: ALN-TTRO2 QPI-1002 Alicaforsen
- 10. 10 • Genes, which are carried on chromosomes, are the basic physical and functional units of heredity • Genes are specific sequences of bases that encode instructions on how to make proteins. • Those proteins that perform most life functions and even make up the majority of cellular structures • When genes are altered so that the encoded proteins are unable to carry out their normal functions GENETIC DISORDERS GENES
- 11. 11 • Insertion of new genetic material into the cells of an individual with the intention of producing benefits for the patients • A technique used to correct defective gene which are responsible for disease development. GENE THERAPY
- 12. 12 • Gene therapy quickly became an intensely investigated field with the promising potential to devise treatment not only for genetic disorders, but also for a wide range of disorders Including metabolic disorders, infectious disorders, chronic illness and cancer. • The treatment of disease by replacing, altering or supplementing a gene that is absent/whose absence or abnormality is responsible for disease.
- 13. 13 • Utilizes gene or short oligonucleotides sequence gene as therapeutics molecules. • Used to treat defective gene which contribute to disease development • Involve one/more foreign gene into an organism to treat hereditary or acquired genetic defect. In gene therapy therapeutic protein which is encoded by DNA is packaged in a vector which transport the DNA inside the cell.
- 14. 14 ADVANTAGES OF GENE THERAPY Gene therapy has the potential to eliminate and prevent hereditary diseases such as: cystic fibrosis, ADA-SCID. It is possible to cure for heart disease, AIDS and Cancer To eradicate disease from future generation Targeted approach: gene therapy allows for precise targeting of specific genes or cells.
- 15. 15 DISADVANTAGES OF GENE THERAPY Immune response: Immune response to the transferred gene stimulates a potential risk to gene therapy Short Lived: Long lasting therapy is not achieved by gene therapy; due to rapid dividing of cells benefits of gene therapy is short lived. Viral vectors: Viruses used as vectors for gene therapy may cause toxicity, Immune response inflammatory reactions in the host Multigene disorder: Heart disease, high BP, Alzheimer's, arthritic and diabetes are hard to treat because it need to introduce more than one gene. Expensive: Making inaccessible for some people
- 16. 16 TYPES OF GENE THERAPY GERMLINE GENE THERAPY (GGT) SOMATIC CELL GENE THERAPY(SCGT) EX VIVO IN VIVO TYPES OF GENE THERAPY
- 18. 18 • In GGT, germ cells are modified by introduction of therapeutic gene • Modifying a germ cell causes all the organism cell to contain the modified gene • The change is therefore heritable and passed onto later generations GERMLINE GENE THERAPY(GGT)
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- 21. 21 SOMATIC CELL GENE THERAPY (SCGT) • In SCGT, the therapeutic genes are transferred into cell other than a germ cell • Introduction of genes into bone marrow cells, blood cells, skin cells • It will not be inherited to later generations • Most focus on severe genetic disorders, including Immune deficiencies, hemophilia, thalassemia and cystic fibrosis, such single gene disorders are good candidates for somatic cell therapy
- 22. 22 SOMATIC CELL GENE THERAPY GERMLINE GENE THERAPY Gene are introduced into somatic cell Gene are introduced to germ line cells and will get distributed in both germ cells and somatic cells Changes are confirmed to the recipient Changes will be passed to the future generations Technical expertise for somatic cells manipulation : introduction of gene of interest and replanting somatic cells in body to make them functional is developed There are still many technical difficulties in introduction of gene into germ cells No moral issue Moral problems to be answered
- 23. 23 EX VIVO & IN VIVO GENE THERAPY 05
- 24. 24 TYPES OF SOMATIC CELL GENE THERAPY EX VIVO IN VIVO
- 25. 25 A.EX VIVO GENE THERAPY • Cells are modified outside the body and then transplanted back in again • Called Exvivo because the cells are treated outside the body Isolate cells with defective gene from a patient Grow the cells in Culture Introduce the therapeutic genes Select genetically corrected cells and grow Transplant the modified cells to the patient
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- 27. 27 ADVANTAGES DISADVANTAGES • Screening of gene expression. • Safety check possible • Close control is possible • Lack of enough cells from target tissues • Defective uptake of gene • Inadequate expression of gene • High production cost ADVANTAGES AND DISADVANTAGES OF EX VIVO GENE THERAPY
- 28. 28 B.INVIVO GENE THERAPY • Direct delivery of therapeutic gene into target cell of patients body. • Called Invivo because the gene is transferred to cell inside the patient’s body. • carried out by viral / non-viral vector systems. • It can be the only possible option in patients where individual cells cannot be cultivated invitro insufficient numbers. • Invivo gene transfer is necessary when cultured cells cannot be re-implanted in patients effectively
- 29. 29 VECTORS FOR IN VIVO GENE THERAPY VIRAL VECTORS SYSTEM NON-VIRAL VECTOR SYSTEM GENE DELIVERY BY VIRUSES GENE THERAPY BY NON-VIRAL VECTORS Many viral vector systems have been developed for gene delivery. These include: • Retro viruses • Adenoviral vector system • Herpes simplex Virus(HSV) Vector system • Pure DNA constructs • Liposomes • DNA molecular conjugates • Human artificial chromosome(HAC)
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- 31. 31 METHODS OF NON-VIRAL VECTOR GENE THERAPY PHYSICAL METHODS GENE GUN MICRO INJECTION ELECTROPORATIO N HYDROPORATION CHEMICAL METHODS USING DETERGENT MIXTURES LIPOFECTION
- 32. 32 • Simplicity: single step vector injection • Minimal invasiveness • To target internal organ ADVANTAGES • Toxicity and immune response issue • Safety check not possible • Binding: vector need to bind efficiently to the cell DISADVANTAGES ADVANTAGES AND DISADVANTAGES OF IN VIVO GENE THERAPY
- 33. 33 IN VIVO EX VIVO Less invasive More invasive Technically simple Technically complex Vectors introduced directly No vectors introduced directly Safety check not possible Safety check possible Decreased control over target cells Close control possible Direct delivery of therapeutic gene into target cell into patients body Isolate cells with defects from patients
- 34. 34 REFERENCES 06 1. An overview of nucleic acid based therapeutic system-Adithya Chaudhary,Nitish Kumar.international journal of pharmaceutical sciences.2023.volume 2, issue 5, 195-205 2. Nucleic acid based gene therapeutics:delivery challenges and molecular design of non-viral gene carriers and expression cassettes to overcome intracellular barrier for sustained targeted expression.Charlie Yu Ming Hsu and Hasan Iludag.journal of drug targeting.2012.volume 4.301-319
- 35. 35 PREVIOUS YEARS QUESTIONS 5 MARKS 1. EX VIVO AND INVIVO GENE THERAPY 2. IN VIVO GENE THERAPY
- 36. 36 THANK YOU
Editor's Notes
- #4: Nucleic acids are polynucleotides. They carry information in cells and make up genetic material Nucleoside sugar+nito Nucleotide sugr+nitro+po4 adenosine guanosine cytidine uridine
- #5: Nitrogen bases DOUBLE(ABZ) AND SINGLE STRAND(RMT) Ag purine cut pyrimidine No u in dna since thymine is more stable resistant to photochemical mutation than u + dna repair enzyme recogonise t than u Dna cell nucleus mitochondria rna cytoplasm nucleus ribosome
- #6: ID: cystic fibrosis(lung and digestive s/s-thick mucus pdction)SICKLE CELL ANEMIA,HEMOPHILIA(prevent clotting),TOURETTE SYNDROME AD:cancer,malaria
- #8: Alicaforsen-antisense(non-coding dna strand) oligont target m rna therapeutics IBD enema Ulceratve colitis crohns disease ICAM 1 pdction Oligo are short dna or rna molecules,oligomer & rnai suppress gene expression ALN(alnylam company)TTR(transthyretin-protein in plasma and csf transport thyroid hormone thyroxine and retinol to liver)PATISIRAN –polyneuropathy with hereditary transthyretin mediated amyloidosis(lipid nanoparticle) Qpi1002(TEPRASIRAN)-short interfencing rna for delayed graft function renal transplant ACUTE KIDNEY INJURY
- #9: most important objective for the medical prospects of gene therapy
- #10: Gene is segment of dna passed from parent to offspring
- #14: ADA SCID adenosine deaminase deficient severe combined immunodeficiency ……damage to immune s/s by mutation of ada gene…impairs development of lymphocytes
- #15: Immune response body immune s/s may see genes as harmful and attack themm
- #17: Germ cell-a cell that develop into a reproductive cell ,egg/ova(oogonia) in female sperm(spermatogonia) male
- #19: Zygote-fertilized egg cell from union of female and male gamete
- #20: cell other than a germ cell Introduction of genes into bone marrow cells, blood cells, skin cells
- #22: Somatic no moral issues bcz it targets only individual patients cell not affect future generation Ggt more controversial potential affect future generation
- #24: Gene expression a process by which information encoded in a gene turned to function Transcription translation
- #26: TRANSFECTION artificially introducing nucleic acid into cells HARVEST MODIFY RETURN
- #27: IMMUNO avoid immune response Screening screen new genetic matrl to ensure its healthy before re-transplantation
- #29: Vectors is a tool to carry a gene into cell CHARACTER:target right cell,activate gene,integrate gene in the cell,no universal vector exist GENE EXPRESSION S/S (VIRAL/NON-VIRAL GENE TRANSFER) RAMYASRI
- #30: Transgen epackaging:process of encapsulating desired transgene into viral vector Transgene-experimently constructed piece of dna
- #31: VIRAL BY IM IV NONVRL Gene gun high pressuredelivery s/sto shoot tissue with gold/tungsten particles that coated with dna Hydro-hydrodynamic pressure to penetrate cell mem (hepatic cell) Detergent=capo4 with fuctional dna Electr-electric field CHEMIICAL LIPOFECTION =lipid:Lnanoemulsion,peptide liposomes
- #33: HARVEST MODIFY RETURN Best approach depends on condition exvivo good in treat blood d,vision d and external disorders Invi INTERNAL ORGAN individual cells cannot be cultivated invitro insufficient numbers. when cultured cells cannot be re-implanted in patients effectively