brief about human therapies-TPA Interferons and Antisense Molecules.

1. Human Therapies- TPA
Interferon and Antisense
molecules.
Submitted By-
Shaista Khan.
MSc.MB.3rd Semester.
A07199319007.
Submitted To-
Dr. Gurjeet Kaur.
Assistant Professor.
AIB .
.

2. Tissue plasminogen
activator -
(tPA)

3. Introduction-
 Serine protease enzyme.
 one of the essential components of the
dissolution of blood clots.
 primary function includes catalyzing the
conversion of plasminogen to plasmin.
 Recombinant biotechnology has allowed tPA
to be manufactured in labs, and these
synthetic products are called recombinant
tissue plasminogen activator (rtPA).
 These drugs have undergone various
modifications to amplify their pharmacokinetic
and pharmacodynamic properties.

4. Some facts about tPA-
 Name:
 Tissue-type plasminogen activator
 Synonyms:
 t-PA, vascular activator, extrinsic plasminogen activator
 History:
 Discovered by Astrup et al 1947
 Synthesis:
 Endothelial cells, mesothelial cells, megacaryocyte
 Molecular weight
 68.000 Dalton
 Carbohydrate:
 7-13%

5. Contd..
 Gene:
 36.6 kb, 14 exons, located at chromosome 8.
 Amino acids:
 527 or 530
 Concentration:
 5 mg/l (antigen)
1 mg/l (activity)
 Half-life:
 4-5 min
 Importance:
 Decreased t-PA activity and elevated t-PA and
PAI-1 antigen may be a risk marker
for cardiovascular disease.

6. Use Of tPA -
 Ischemic stroke (most common).
 Myocardial Infarction.
 Pulmonary embolism.
 Thrombolysis (e.g., deep vein
thrombosis).

7. Sequences of the action of
tPA -
 tPA attaches to the fibrin on the clot
surface.
 It activates the fibrin-bound
plasminogen.
 Plasmin is subsequently cleaved from
the plasminogen affiliated with the
fibrin.
 The molecules of fibrin are broken up
by the plasmin, and the clot dissolves.

8. INTERFERONS
-

9. What are Interferons-
 In 1957, Isaac and Lindenmann
discovered a substance that interfered
with viral replication.
 Naturally occuring proteins and
glycoproteins.
 Structurally, they are the part of
cytokine family.
 Secreted by Eukaryotic cells in
response to viral infections, tumors
and other biological inducers.

10. Important Features-
 Part of non-specific immune system.
 Play an important role in the first line
of defense against viral infection.
 Itself is not an anti-viral agent.
 By inhibiting viral replication, it induces
antiviral state.

11. Types Of Interferon-
 Type 1 (Interferon alpha and beta)-
 Produced when the body recognizes a
virus that has invaded it.
 Produced by fibroblast and monocytes.
 These IFNs binds to specfic receptors on
target cells which lead prevent the virus
from replicating its genetic material.
 IFN-alpha used to treat hepatitis B and
C.
 IFN-beta can be used to treat multiple
sclerosis.

12. Contd..
 Type II (Interferon gamma)-
 Also known as immune Interferon and
activated by Interleukin-12.
 Released by cytotoxic Tcells and type-
1 T helper cells.
 Results in inhibition of Type-2 helper
cell’s immune response and induction
of Type-1 helper cell’s immune
response.
 These Interferons binds to Interferon
gamma receptor (IFNGR).

13. Contd..
 Type III-
 Signal through a receptor complex
consist of IL10R2 and IFNLR1.
 Although discovered more recently
than type I and type II IFNs.
 Information about these types of
Interferons are demonstrated in some
virus and fungal infections.

14. Functions Of Interferons-
1-Antiviral agents and modulate functions
of immune system:-
 In response to IFNS, cells produce large
amount of enzyme (protein kinase R).
 Phosphorylates elF-2 which further forms
an inactive complex with elF2B protein to
reduce protein synthesis within the cell.
 Also Interferons induce production of
hundreds of other proteins (Interferon-
stimulated genes).

15. Contd..
2- Up-regulate major histocompatibilty
complex molecules and increses
immunoproteosome activity:-
 Higher MHC I expression increases
presentation of viral and abnormal
peptides to cytotoxic T cells, where as
MHC II present these peptides to T
helper cells.
 Immunoproteosome processes peptides
to load on MHC I molecule.
 Thus, increases the recognition and
killing of infected cells.

16. Contd..
3- can suppress angiogenesis by down
regulation of angiogenic stimuli.
4- supresses the proliferation of
endothelial cells.
 Cause decrease in tumor
angiogenesis.
 Decreases its vascularization.
 IFN type III, activates other immune
cells, like macrophages and NK cells.
 Inhibits cell growth.

17. Viruses resistance to Interferons-
 Blocks the downstream signaling and
prevent further IFN production and also
inhibit the functions of proteins induced
by IFN.
 H5N1 influenza virus, has resistance to
interferon and other-viral cytokine.
 Encodes proteins that binds to dsRNA to
prevent the activity of RNA-dependent
protein kinase.
 Eg. JEV, dengue type 2 virus, SARS-
COV-2, etc.

18. Interferon Therapy-
 Therapy is used as a treatment for some
cancers.
 Treatment can be used in hematological
malignancy, such as leukemia and
lymphomas.
 Those treated with Interferon have a
sustained virological response and can
eliminate hepatitis virus.
 Biopsies of patients given the treatment
show reduction in liver damage and
cirrhosis.
 In systemic therapy, IFNs are
administered by an intramuscular
injection.

19. Antisense
Molecules-

20. Definition-
 The term antisense molecules comprises
several classes of oligonucleotide
molecules that contain sequence
complementary to target RNA molecules,
such as mRNA, viral RNA, or other RNA
species, and that inhibit the function of
their target RNA after sequence-specific
binding.
 4 major classes- antisense
oligodeoxyribonucleotide (ssDNA
molecules), ribozyme, small interfering
RNA molecules, and DNAzymes.

21. General features of antisense
molecules families-
ssDNA molecule Small interfering
RNA molecule
Ribozyme DNAzyme
size 12-25 bases 21-25 base pairs Lesser than 30 bases
(hammerhead
RNAzymes)
Lesser than 50 bases
(hairpin RNAzymes)
30-35 bases.
oligo type and
structure
Linear ssDNA Linear dsRNA Complex RNA with
single and double
strand sections.
ssDNA with 2
binding domains
surrounding a central
catalytic domain.
Inherent Enzymatic No No Yes Yes
Advantages Easy to produce and
modify.
Degradation of whole
mRNA
Can be expressed
from coding DNA
sequences in vivo.
Good in vivo cell
penetration.
Disadvantages Off-target effects. Difficult to produce. large size and
complex structure.
Activity depends on
intracellular Mg2+
level

22. Applications-
 Used application of therapeutic
antisense molecules is to reduce
expression levels of proteins associated
with central mechanisns of disease
development.
 Can also be developed to interfere with
other functional aspects of RNA
molecules.
 Mode of action of antisense molecules
revealed many of its structure involved
enzymatic activities to fully exert their
function.

23. Use in inflammatory
disorders-
 First antisense molecules applied in
clinical studies was alicaforsen (ISIS
2302).
 Combination of 2 antisense DNA
molecule called TPI ASM8 was
developed for the treatment of allergic
asthma.
 Most recently, a DNAzyme-based drug
was successfully applied in clinical study
for the first time.
 SB010 represents a DNAzyme directed
against GATA-3, the master transcription
factor of TH2-driven immune responses.

24. Thank you