NEW ERA OF TREATING CANCER

1. ARE JAK 2 INHIBITORS THE SOLUTION
FOR TREATMENT OF
MYELOPROLIFERATIVE DISEASES

2. DONE BY:
TAMMALI HARIKA (09Z31R0020)
AINALA SRAVANI REDDY (09Z31R0024)
KUTHADI PRIYANKA (09Z31R0042)
MAILARAPU NAVYA (09Z31R0045)
MUTHYAM ROJA RANI (09Z31R0046)
SAFURA AYESHA MUJEEB (09Z31R0054)
UNDER THE GUIDANCE OF
MRS . SIVA JYOTHI BUGGANA,
M. PHARMACY
MEDICINAL CHEMISTRY

3. CONTENTS.
• Abstract
• Introduction to cancer
• Targets of Cancer
• Plan of work
• Introduction to Janus Kinases (JAK)
• Introduction to STAT
• Coordination between JAK and STAT
• Scaffolds used to synthesize JAK2 inhibitors
• JAK2 inhibitors under clinical phases
• Conclusion
• Bibliography

4. • ABSTRACT:
• Janus kinases (JAKS) are critical components of cytokine signaling
pathways which regulate immunity, inflammation, haematopoiesis,
growth and development.
• The recent discovery of JAK2- activating mutations as a casual event in
majority of patients with Philadelphia chromosomes negative (ph-
)myeloproliferative disorders (MPDS) promoted many pharmaceutical
companies to develop JAK2 selective inhibitors for the treatment of
MPD’S

5. • JAK2 inhibitors effectively reduce JAK2 driver phosphorylation of signal –
transducer and activator of transcription 5, and cell survival in JAK 2
activated cells in vitro and invivo.
• Patients treated with these inhibitors experience a rapid reduction of
splenomegaly, significant improvement of constitutional symptoms.

6. INTRODUCTION:
• Cancer is a disease in which there is uncontrolled multiplication
and spread within the body of abnormal forms of the body’s own
cells.
• They are distinguished from benign tumours by the properties of
dedifferentiation, invasiveness and the ability to metastasise (spread
to other parts of the body) both benign and malignant tumours
manifest uncontrolled proliferation.

7. • PATHOGENISIS OF CANCER:-
• Cancer cell distinguish from normal cell by :-
• 1. Uncontrolled proliferation
• 2. Dedifferentiation
• 3. Invasiveness
• 4. Metastasis.

8. APOPTOSIS
• Apoptosis is programmed cell death . Development of resistance to
apoptosis is hallmark of cancer. The decrease of apoptosis can be
brought about inactivation of proapoptotic factors or by activation of
antiapoptotic factors

9. GENESIS OF A CANCER CELL:-
A normal cell turns into a cancer cell because
of one or more mutations in its DNA which can
be inherited or acquired.
There are two main categories of genetic change
that lead to cancer they are :
1)Activation of proto oncogenes to oncogenes .
2)Inactivation of tumour suppressor genes.
Eg: breast cancer.

10. CELL CYCLE:-
The normal cell proliferate throught a definite cell cycle that causes
programme DNA replications.
The mechanism of cell division is same in all dividing phase and is divided
into 5 phases.
They are:
1. Go(Gap o (or) Resting (or) non-Proliferative phase)
2. Phase G1
3. S Phase
4. G2 Phase
5. M-Phase

12. NEGATIVE REGULATORS OF CELL CYCLE:-
• Rb protein that holds the cycle in check while it is hypophosphorylated
another negative regulatory mechanism is the action of the inhibitors of
the cdk’s.
• These bind to and inhibits the action of the complexes, their main actions
being at check point 1.
THERE ARE 2 FAMILIES OF INHIBITORS:-
1) Kinase inhibitory protein family (cdk inhibitory proteins, also termed
KIP/kinanes inhibitory proteins:P21,P27 and P53.
2) Inhibitors of kinase family (INK FAMILY):- P16, P19, P15.

13. TARGET SITES
• A) CONVENTIONAL OR TRADITIONAL TARGETS:
• These include alkylating agents, anti metabolites, antibiotics, plant
alkaloids, radio isotopes.
• B) NEXT GENERATION TARGETS:
• 1. Cell signaling pathway
• 2. Protein therapy
• 3. Protein- protein interaction
• Cancer related genes as targets:
• These genes are of 2 types they are:
• a)oncogenes. b) Tumour suppressor gene.

15. 1.Oncogenes:
oncogenes contribute to the transformation process by driving cell
proliferation or reducing sensitivity to cell death.
Classes of oncogenes-
a)Growth factors: GF’S are produced by one cell and they act on other cells
this is known as paracrine stimulation. Cancer cell secrete their own GF’S
and they proliferate to form tumours this is known as autocrine
stimulation.
b)Growth factor receptors
c)Membrane associated guanine nucleotide binding protein.
d)Serine threonine protein kinases.
e)Cytoplasmic proteins.
f)Cytoplasmic tyrosine kinases.

16. 2.Tumour suppressor genes:
These genes can directly or indirectly inhibit cell growth.
Direct inhibitors promote cell death and are called `gate keepers’ and
their activating is rate limiting for tumor cell proliferation. Indirect
inhibitors promote genetic stability and are called `care takers’ which
function in DNA repair pathways and whose elimination causes
increased initiation rate.
a)Retinoblastoma : Loss of Rb protein function leads to malignancy. It may
be either phosphorylated or unphosphorylated.
b)p53: it is activated in respose to cellular stresses created by activated
oncogenes.

18. Biochemical targets:
a)Reactive oxygen species- Ionizing radiation is the complete carcinogen and
causes DNA damage.
b)Matrix metalloproteinases -They play an integral role in malignant
progression.
c)Telomerase expression-Telomers are specialised structures that cap the ends
of chromosomes (like small metal tubes on the end of shoe lace)
protection them from the degradation, rearrangement and fusion with
other chromosomes. A portion of telomere is eroded with each round of
cell division so that eventually it becomes non functional

19. PLAN OF WORK
• Till now anticancer drugs available in the market are almost having
same mechanism of action, that is damaging the DNA synthesis in
affected cells and also damaging the normal cells along with the
neoplastic cells.
• Now we are trying to collect the information regarding the
treatment of cancer by using future generation targets, one of that is
Janus kinase inhibitors. Here we are ready to present the information
regarding Janus kinases.

20. INTRODUCTION:-
 JAK2V617F was the first mutation to be described in patients with ph-
negative MPN’S and the fact that it can be detected in patients with all 3
classic subtypes of MPN’s.
 After discovery of JAK2V617F;other investigators discovered mutation
in related genes of hemopoietic growth factor signaling
pathways,including exon 12 mutations in JAK2,Mutation of MPL.

21. THE JAK FAMILY OF TYROSINE KINASE:
• The JAK family of tyrosine kinase (named after the two faced roman god
janus) was first described in 1989.
• There are four members of the JAK family ; JAK1,JAK2,JAK3 and TYK.
• JAKs are cytoplasmic kinases that associate with the intracellular portion
of cytokine receptors that do not posses intrinsic kinase activity ,such as
receptors for hematopoietic growth factors (erythropoietin receptor [EPOR]
,G-CSF receptor [G-CSFR] ,and thrombopoietin receptor [C-MPL]
• Binding of the putative ligand leads to receptor dimerization and
subsequent approximation of two JAK kinases ,which transphosphorylates
and activate each other, initiating intracellular signalling pathways

23. • One of the most important intracellular signalling pathways activated by
JAKs is the JAK-STAT(signal transducer and activator of transcription)
pathway.
• STAT are latent,cytoplasmic transcription factors. JAKS phosphorylate
STATs tyrosine residues; leading to STAT dimerization , translocation to
the nucleus and activation of transcription.
• Aberrant activation of STAT3 and STAT5 has been linked to neoplastic
transformation.

24. • Other signalling pathways which can be activated by ;JAKs include the
Ras/Raf/MAPK pathway and the PBK/Akt pathway. Activation of these
pathways leads to increased cellular proliferation and resistance to
apoptosis , and deregulation could cause the development hematological
malignancies.
• Structurally,JAKs consists of seven different domains

25. THE JAK 2 V617F MUTATION:-
• It was identified primarily in patients with PV (90-97%), ET (60%) and
PMF (50%).
• It was identified primarily in patients with PV (90-97%), ET (60%) and
PMF (50%).
• In PV > 75% allelic burden is associated with higher hematocrit, white
blood cell (WBC) count, and higher incidence splenomegaly and
thrombotic events. Patients with primary MF who are JAk 2 V617F –
positive may have higher haemoglobin & WBC count and a higher risk of
leukemic transformation & of developing massive splenomegaly . V617F
appears to promote a polycythaemic phenotype in patients with both ET &
IMF

26. • The patients with V617F – negative MPNS led to the discovery of other
mutation JAk 2 exon 12 mutation in 3% of pv patients.
• These mutations also lead to increased cellular proliferation and hyper
sensitivity or independency to hematopoietic growth factors indeed, JAK 2
V617F negative erythroid colonies from patients with PV can grow in the
absence of erythropoietin.
RATIONALE FOR TARGETING JAK 2 IN MPN’S :
• Imatinib, These drugs target kinases that are abnormally activated in
cancer cells, with the objective of blocking cellular proliferation and
inducing apoptosis .

27. There is great therapeutic benefit for employing these drugs to achieve
symptom control and improve the quality of life of patient with MPNS

28. SCAFFOLDS USED AS JAK-2 INHIBITORS:
QUINAZOLINES 2,4-diamino triazole.
QUINOXALINES 3,4-ring fused 7-
azaindoles

29. Pyrazole-3-ylamino pyrazines Thienopyridines
Pyrazolo[1,5-a] pyrimidines 2-thio-benzoxazoles

30. JAK2 INHIBITORS UNDER CLINICAL TRIALS
1. CEP-701: It is also known as a lestaurtinib is a TK1 which belongs to the
chemical class of indocarbazol of alkaloid. It is a potent FLT3 and JAK2
inhibitor.
• DOSE: 80-100mg twice daily as a liquid formulation.
• USES:
 Reduction in spleen size with improvement in cytopenias.
 Use to treat patients with JAK2V617F-positive primary MF (or) post
PV/ET MF and clinical .
• ADRS:
* Myelosuppression.
*GI toxicity

31. 2. AZD1480: it is a pyrazolyl pyrimidine compound that selectively and
potently inhibits JAK2.
• MOA: This compound block STAT 5 activation, inhibit the cell
proliferation and induced apoptosis in the human JAK2V617F positive.
• In a mouse model, AZD-1480 reduced the proliferation of stem cells
transferred with the JAK2 mutant protein.
• USES:
 Therapy with AZD 1480 inhibited tumour growth, and tumour cells
lysates had reduced level phospho STAT3.
 A phase 1 clinical trail is currently underway to evaluate the activity of
AZD1480 in patients with MPNS.

32. 3.R723: This compound is strongly antiproliferative against mouse and
human cell lines that harvor the JAK2V617F mutation , constitutive
STAT5 phosphorylation in JAK2V617F positive cells was inhibited 10-20
fold more potently than IL-2 induced STAT5 phosphorylation
4. XL019: It is a potent , reversible and selective inhibitor of both wild
type and mutated JAK2 and shows good selectivity for JAK2 invitro
biochemical assays.

33. DOSE: Given orally once daily or three times weekly.
Tolerated dose is 25 and 50mg/kg.
ADRS:
* Non-haematological adverse effects.
* Leukocynosis, decrease of circulating blasts, improvement of anaemias,
pruritis and poor appetite.

34. 5. INCB018424: It is a potent pan JAK inhibitor targeting JAK1,
JAK2,and TYK2.
• MOA:INCB018424 Inhibits constitutive and IL-6 stimulated
phosphorylation of STAT3 and reduces production of proinflammatory
cytokines in all MF patients regardless of their JAK2 mutation status.
• DOSE: The starting dose of 25mg pobid was demonstrated to be
maximum tolerated dose.
• USES:
 Therapy with INCB018424 also improved hyper catabolism-associated
hypocholestrolemia.
 INCB018424 treatment also resulted in a rapid reduction of spleenomegaly.

35. INCB-18424
• 6. SB1518: It is a JAK2 inhibitor that has activity against both wild
type and mutated JAK2,being selective against JAK1 and JAK3.
• MOA: SB1518 INHIBITS PROLIFERATION OF Baf/3 cells
transferred with the erythropoietin receptor (EPOR) and the JAK2
V617F mutation and decreased phosphorylation of JAK2 and STAT5.

36. • DOSE: Ranged from 100- 600mg orally once daily in 28-days cycles.
• ADRS:
 Abdominal pain and diarrhea.
 Nausea and thrombocytopenia.
7. TG101348:It is an orally available, potent JAK2 inhibitor with greater
selectivity against JAK1 and JAK3.

37. . USES:
• TG101348 is able to induce response in splenomegaly in the majority of
patient with MF and this may be due to a direct effect on inhibition of
JAK2V612F.
• MOA:TG101348 inhibited proliferation and induce apoptosis of JAK2
V617F positive HEL cells and Ba/f3 cells transduced with JAK V617F.
Dose: TG101348 was well tolerated in patience with MF when the drug was
taken orally once a day.
ADRS:
Non hematological toxicities.
Thrombocytopenia, neutropenia, Tronuition depended anaemia.

38. • TG101348 also led to improvement in system is symptoms, including
pruritis, night sweats, fatigue
• 8. CYT387:It is a amino pyrimidine compound is an ATP competitive
small molecules that potently inhibits JAK1 and JAK2 . CYT387 also
inhibits JAK3 but to a lesser extent.
MOA:CYT387 inhibits proliferation of
cell lines that depend on signaling by
JAK kinases for proliferation, including
Ba/f3 cells engineered to express both
EPOR and JAK2 V617F .

39. • USES:
 Therapy with CYT387 in mice developed with the splenomegaly ,
erythrocytosis , leukocytosis , splenomegaly and bone marrow fibrosis
improved hemoglobin levels , normalized WBC counts and reduced spleen
size.
 A reduction but not elimination of JAK2 disease burden , with partial
normalization of progenitor cell distribution and differentiation by this
compound was also observed in the model.

40. • DRUGS UNDER CLINICAL TRIALS
•
DRUG CURRENT PHASE
CEP701 ‫װ/׀‬
AZD1480 ‫׀‬
R723 Pre-clinical
XL019 ‫׀‬
INCB18424 ‫׀׀׀‬
SB1518 ‫׀‬
TG101348 ‫װ‬
CYT387 ‫׀‬

41. Conclusion
• Jak 2 inhibitors are a new class of drugs being actively developed for
patients with ph negative MPNs they are the solution to all clinical
issues in patients with MPNs, great clinical benefit can be obtained by
the use of these drugs.
• In patients with MF, therapy JAK2 inhibitors can lead to improvement
in spleen size, WBC count weight loss, fatigue, night sweats and is
associated with improved quality of life.

42. BIBILOGRAPHY:-
1. Baxter EJ, Scott LM, Campbell PJ, et al. Acquired mutation of the
tyrosine Kinase JAK 2 in human myeloproliferative disorders, Lancet 2005;
365: 1054-61.
2. James C; Ugo V, Le Couedic JP et al. A unique clonal JAK 2 mutation
leading to constitutive Signaling causes polycythaemia Vera, Nature
2005; 434: 1144-8.
3. Kralovics R, passamonti F, Buser AS, et al; A gain of function mutation
of JAK 2 in myeloproliferative disorders N Engl Med 2005; 352: 1779-90.
4. Levine RL, Wadleigh M, Cools J, et al. Activating mutation in the
tyrosine kinase JAK 2 in polycythemia vera, essential thrombocythemia
and myeloid metaplasia with myelofibrosis, Cancer cell, 2005; 7: 387-97.

43. • 5. Akada H, Yan D, Zou H, et al. conditional expression of heterozygous
(or) homozygous JAK 2 V 617 F from its endogenous promoter
induces a polycythemia Vera like disease Blood 2010; 115: 3589-97.6.
Bumm TG; Elsea C,
• 6. Bumm TG; Elsea C, Corbin AS, et al. characterization of murine JAK 2
V 617F-positive myeloproliferative disease Cancer Res 2006; 66: 11156-65.
• 7. Lacout C, pisani DF, Tulliez M, et al. JAK 2 V 617 F expression in
murine hematopoietic cells leads to MPD mimicking human pv with
secondary myelofibrosis, blood 2006; 108: 1652-60.
• 8. LiJ, Spensberger D, Ahn JS, et al, JAK 2 V 617 F impairs hematopoietic
stem cell function in a conditional Knock-in mouse model of JAK 2 V 617
F-positive essential thrombocythemia, Blood 2010; 116: 1528-38.

44. • 9. Marty C, Lacout C, Martin A, et al. myeloproliferative neoplasm
induced by constitutive expression of JAK 2 V 617 F in Knock-in mice,
Blood 2010; 116; 783-7.
• 10. Mullally A, Lane SW, Ball B, et al. physiological JAK 2 V 617 F
expression causes a lethal myeloproliferative neoplasm with differential
effects on hematopoietic stem and progenitor cells. Cancer cell 2010;
17:584-96.
• 11. Shide K; Shimoda HK; Kumano T, et al. Development of ET,
primary myelofibrosis and pv in mice expressing JAK 2 V 617 F;
Leukaemia 2008; 22: 87-95.

45. • 12. Tiedt R, Hao-Shen H, Sobas MA, et al. Ratio of mutant JAK 2 V
617 F to wild- type Jak 2 determines the MPD phenotypes in transgenic
mice. Blood 2008; 111:3931-40.
• 13. Wernig G, Mercher T, Okabe R, et al. Expression of JAK 2 V 617 F
causes a polycythemia vera-like disease with associated myelofibrosis in
a murine bone marrow transplant model. Blood 2006; 107: 4274-81.
• 14. Xing S, Wanting TH, Zhao W, et al. Transgenic expression of JAK
2 V 617 F causes myeloproliferative disorders in mice. Blood 2008;
111: 5109-17.
• 15. Zaleskas VM, Krause DS, Lazarides K, et al. Molecular
pathogenesis and therapy of polycythemia induced in mice by JAK 2 V
617 F. PLOS one 2006; 1: e18.

46. THANK YOU