kinase inhibitors for cancers

8/8/2019 Kinase Inhibitors for Cancers

1/55

Kinase Inhibitors for cancertherapy

Maulik P. Suthar

Feb 2009Department of BiotechnologyShree S. K. Patel College of Pharmaceutical Educationand Research, Ganpat University, At: Kherva, PIN-

382711, Gujarat, India


2/55

Human Protein kinases (PKs)

TKL

STE

CKI

AGCCAMK

TK

CMGC


3/55

Why target Protein kinases?

Protein kinases (PKs) mediates substrates

phosphorylation PKs are indispensable for numerous processes

Under pathological conditions PKs can be

deregulated, leading to alterations in thephosphorylation and resulting in uncontrolledcell division, inhibition of apoptosis

Some protein kinase inhibitors currently undergoclinical trials or have already been successfullyintroduced into treatment


4/55

Important Kinases for drug discovery

Bcr-Abl Abelson tyrosine kinase

PKC Protein kinase C CDK Cyclin dependent kinase ERK Extracellular signal-regulated kinase

JNK c-Jun N-terminal kinases MAPK Mitogen-activated protein kinase MKK MAP kinase

PDK-1 Phosphoinositide-dependent kinase-1 PI3K Phosphatidylinositol 3-kinase PK Protein kinase

PKC Protein kinase C RPTK Receptor protein tyrosine kinase


5/55

Mechanism of action

Molecules have alow molecularweight and mostof them bind toprotein kinasescompeting withATP for the ATP-binding site


6/55

ATP binding pocket

Interaction of CDK2 with ATP. Dotted lines indicate the Van Der Waals contacts.

Thick broken lines indicate the Hydrogen bonds. Atoms outside the specific aminoacid box with solid line indicate the interaction participation with particular ATP atom.(Kim S. H. et al, (1998))


7/55

Protein kinase inhibitors

Protein kinase inhibitors (PKIs) are chemicallydiverse, low-molecular-weight, less than 600 Da,hydrophobic heterocycles. While most PKIscompete with the ATP substrate, there alsoexists a group of the

ATP non-competitive inhibitors, which have beendescribed as a group of peptide inhibitors ofprotein kinases

More than 30 ATP-competitive inhibitorscurrently undergo clinical trials.


8/55

Kinase assays

The potency of a PKI is typically expressed as

the IC50 value concentration of the drug atwhich 50% of kinase activity is inhibited. Mostkinase inhibitors are reversible, and their IC50

depends on the dissociation constant of theinhibitor and ATP concentration


9/55

Tyrosine Kinases (TKs)

Receptor TKs:

c-kit, insulin-like growth factor receptor, EGFR, Vascular EGFR (VEGFR),

Fibroblast growth factor receptor (FGFR),

Platelet-derived growth factor receptor(PDGFR).

Ligand-mediated activation of VEGFR by VEGFsecreted by tumour cells, which provides thetumour vascularization


10/55


11/55


12/55

Tyrosine Kinases (TKs)

Cancers : breast cancer, amyotrophic lateralsclerosis

Inhibitors : PKC412,

SU11248, PTK787,

Gleevec +SU5416,

sorafenib


13/55


14/55

Bcr-Abl kinase

Reciprocal recombination occurs between bcr

andabl

genes. Because of an increased tyrosine kinase activity,

Bcr-Abl causes cell growth and differentiation

and reduces apoptosis.


15/55


16/55

Bcr-Abl kinase


17/55

Bcr-Abl kinase inhibitors

Cancers : CML

Inhibitors: imatinib mesylate

(Gleevec, STI571),

BMS-354825

(dasatinib),

VX-680


18/55

CDK inhibitors

CDK inhibitors are a heterogeneous group of

compounds that are able to inhibit CDKsinvolved in the cell cycle (CDK1, CDK2, CDK3,CDK4, CDK6, and CDK7), transcription (CDK7,

CDK8 and CDK9), or neuronal functions (CDK5and CDK11).


19/55

CDK inhibitors


20/55

CDK inhibitors

Cancers : various types of sarcomas, colorectal andlung cancers

Inhibitors: flavopiridol roscovitine purvalanol B

olomoucine UCN-01 E7070

BMS-387032 purvalanol A (P 4484) kenpaullone (K 3888)

alsterpaullone (A 4847) indirubins staurosporine (S 4400)


21/55

EGFR inhibitors

EGFR inhibitors target the intracellular domain ofthe receptor TK competing with ATP for theintracellular catalytic site of EGFR and thusblock its downstream signalling.

Therefore, they inhibit tyrosineautophosphorylation, resulting in a blockade ofEGFR signal transduction pathways.


22/55

EGFR inhibitors

Overexpression, point mutations in the kinase

domain or both lead to different cellularprocesses involved in carcinogenesis such ascell proliferation, inhibition of apoptosis,

angiogenesis, cell motility, and metastasis.


23/55


24/55

EGFR inhibitors


25/55

EGFR inhibitors

Cancers: colorectal cancer, non-small-cell lung

cancer, glioblastoma multiforme, different typesof solid tumours

Inhibitors:

erlotinib,

gefitinib (Iressa),

PKI166, PD153035,canertinib


26/55

JAK inhibitors

Janus kinase (JAK) family members the mainachievements have been made in inhibition of

JAK3 and JAK2. JAK3 inhibition blocks several cytokine signals in

NK cells and in T and B lymphocytes. It can

provoke immunosuppression by altering theexpansion and function of these cells. Therefore, targeting JAK3 may theoretically be

used for immune suppression where it isneeded, e. g. on cells actively participating intransplant rejection without affecting any cellsoutside of these cell populations


27/55

JAK inhibitors


28/55

JAK inhibitors

1. Deficiency of JAK3

2. A clonal somatic mutation in the pseudo-kinasedomain JAK2

3. Aberrant activity of the JAK-Src kinase duet

JAK i hibi


29/55

JAK inhibitors

Cancers : haemopoietic abnormalities including

leukaemia and SCID , polycythemia vera Inhibitors:

CP-690 550

AG-490 WHI-P131 WHI-P154

A77 1726

MAPK ki


30/55

MAPKs kinase

Superfluous endothelial cell activation, T-effectorcell differentiation and proliferation of vascular

smooth muscle cells Cancers: diabetes, atherosclerosis, stroke,

Parkinsons disease, Alzheimers disease,

arthritis, asthma Inhibitors:

roscovitine,

olomoucine, purvalanols,

PD98059

MAPK ki


31/55

MAPKs kinase

PKC ki


32/55

PKC kinase

1. Anti-apoptotic signalling

2. Promotion of the expression of cell surfacereceptors including the EGF receptor

Cancers: GISTs, breast cancer, different

malignancies Inhibitors:

LY317615 PKC412

PKC ki


33/55

PKC kinase

A rora A and A rora B kinase


34/55

Aurora A and Aurora B kinase

Overexpression of both Aurora A and B

Gene amplification of Aurora A Cancers: breast, bladder, gastric and colorectal

cancers

Inhibitors:

ZM447439,

hesperadin, VX-680



35/55


The signal transduction cascade leading from activation of the progesterone receptor (PR) byprogesterone, through the activation of Aurora A kinase, and the influence of Aurora A kinase andXGef on early CPEB activation. CPEB then participates in the polyadenylation induced translationof c-mos mRNA, which triggers the activation of mitogen activated protein kinase (MAPK).Activated MAPK, in conjunction with polyadenylation-induced translation of cyclin and Cdc25activation (not shown) stimulates the timely activation of MPF (cyclin B: cdc2), which subsequentlytriggers resumption of meiosis. Arrows indicate positive feedback pathways that further stimulatec-mos mRNA translation.

Src kinase


36/55

Src kinase

Deregulation of multiple oncogenic pathwaysincluding PDGFR, VEGFR, and others

Cancers: myeloproliferative disorders, gliomas,carcinomas, melanomas and other malignancies

Inhibitors: dasatinib

Src kinase


37/55

Src kinase

Src kinase


38/55

Src kinase

Src kinase


39/55

Src kinase

Src kinase


40/55

Src kinase

Progression through the cell cycle is accompanied by activation of the proto-oncogene c-Src, a protein tyrosine kinase. Overexpression of Src leads to tyrosinephosphorylation of multiple protein substrates and cellular transformation. Duringinterphase the Src protein folds back upon itself to stay in the inactive state, with a

phophotyrosine residue in one domain at Tyrosine 529 bound by an SH2 domain inthe same protein. Activation of c-Src involves protein-tyrosine phosphatase alpha(PTP-alpha, or RPTP-alpha), a transmembrane protein with a cytoplasmicphosphatase domain. A variety of evidence has indicated that PTP-alphadephosphorylates c-Src at Tyr529, allowing Src to open up and become activated,and that this activation occurs in association with mitosis. To activate Src, PTP-alpha

must first open up the folded Src through binding itself to the phosphorylated Srcdomain, a process blocked by binding of Grb-2 to PTP-alpha at phosphorylatedTyr789. PTP-alpha phosphorylated at Tyr789 also binds to the Src SH2 domain,causing the Src structure to open at Src Tyr529 to become available fordephosphorylation. During mitosis the mitotic kinase Cdc-2 phosphorylates Src, alongwith other cellular substrates, and in so doing makes Src more prone PTP-alpha

dephosphorylation and activation. The activity of PTP-alpha toward Src is alsoregulated by phosphorylation of PTP-alpha by protein kinase C at serines 180 and204, releasing the inhibition of PTP-alpha by Grb-2. In the normal cell cycle, Srcactivity is down-regulated after cell division through dephosphorylation by proteinphosphatases and phosphorylation by Csk (C-terminal src kinase) and PTP-alphadephoshorylation returns the cycle to its interphase condition. The regulation of Src

activity during mitosis demonstrates how protein phosphorylation can shifts thedelicate equilibrium of molecular interactions and cellular responses

PDGFR kinase


41/55

PDGFR kinase

Activating mutations resulting in uncontrolled cellproliferation and maintenance of tumour blood

vessels Cancers : myeloproliferative disorders, gliomas,

carcinomas, melanomas, sarcomas, GIST,

breast and lung cancers, ovarian tumours Inhibitors:

imatinib

PKC412

SU11248

MLN518

PTK787

sorafenib

PDGFR kinase


42/55

PDGFR kinase

Plk/Plk-1 kinase


43/55

Plk/Plk 1 kinase

Deregulation of cell cycle progression

Cancers : head and neck cancer, ovariancancer, endometrial cancer, prostate cancer,NSCLC, glioma, breast cancer, melanoma,

colorectal cancer Inhibitors:

scytonemin

Plk/Plk-1 kinase


44/55

Plk/Plk 1 kinase

ROCK kinase (Rho kinase )


45/55

ROCK kinase (Rho kinase )

Contribution to inhibition of apoptosis in tumourcells

Involvement of the ROCK pathways in motilityand invasion of tumour cells

Cancers : glioma, NSCLCs and cardiovasculardisorders

Inhibitors:

Y27632

Y-30141

ROCK kinase


46/55

ROCK kinase

Flt-3 kinase


47/55

Flt 3 kinase

Mutations leading to constitutive activation of Flt-3, which enhance cell proliferation,

differentiation, and survival Cancer: various haematologic malignancies

incl. acute myeloid Leukaemia

Inhibitors : AG1295 AG1296 MLN518

SU5416 PKC412 CEP-701 SU11248 Ki23819

Flt-3 kinase


48/55

Flt 3 kinase

Flt-3 kinase


49/55

Flt 3 kinase

Flt-3 kinase


50/55

t 3 ase

Transcription of the FMS-like tyrosine kinase 3 (FLT3)gene produces FLT3mRNA, which is translated to FLT3protein. FLT3 contains five extracellular immunoglobulin-like domains (E), a transmembrane domain (TM), ajuxtamembrane domain (JM) and two tyrosine-kinasedomains (K) that are linked through the tyrosine-kinaseinsert (KI). Cytoplasmic FLT3 undergoes glycosylation(G), which promotes localization of the receptor to themembrane. Wild-type FLT3 remains as a monomeric,inactivated protein on the cell surface until FLT3 ligand(L), probably in a dimeric form, binds the receptor and

induces receptor dimerization. FLT3 dimerizationpromotes phosphorylation (P) of the tyrosine-kinasedomains, thereby activating the receptor anddownstream effectors. The dimerized receptors are

quickly internalized and degraded.

c-kit kinase


51/55

Gain-of-function mutations leading to thepermanent activation of c-kit signalling in the

absence of binding of SCF, which leads touncontrolled cell proliferation and resistance toapoptosis. ligand-mediated activation of kit

Cancer: GISTs, lung cancers, Merkel cellcarcinoma, Kaposis sarcoma, germ celltumours, mast cell tumours, melanoma,testicular and gynaecological cancers,

neuroblastoma Inhibitors :

imatinib

SU5416 PKC412 MLN518

c-kit kinase


52/55

Bub1,BubR1,Mps1


53/55

, , p

Mutation followed by spindle assemblycheckpoint and cytokinesis deregulation

Cancer: colorectal cancer

Inhibitors : not available

ATP non-competitive proteine kinaseinhibitors


54/55

inhibitors

ATP competitive PKIs have a drawback theymust compete with high intracellular ATP

concentrations. To be specific these inhibitors must discriminate

between the ATP-binding sites resembling in

multiple human proteins that also utilize ATP,including other PKs.

Therefore, it may be beneficial to target sites onprotein kinases other than the ATP-binding sitedistinct in different PKs

Resources


55/55

Kinase-Disease Associations:

http://www.cellsignal.com/reference/kinase_disease.html

Measuring ATP by bioluminescence method:

http://www.promega.com/multimedia/bioLum01.htm

Bio-pathwayshttp://www.biocarta.com/
http://www.cellsignal.com/reference/kinase_disease.htmlhttp://www.cellsignal.com/reference/kinase_disease.htmlhttp://www.promega.com/multimedia/bioLum01.htmhttp://www.promega.com/multimedia/bioLum01.htmhttp://www.biocarta.com/http://www.biocarta.com/http://www.promega.com/multimedia/bioLum01.htmhttp://www.promega.com/multimedia/bioLum01.htmhttp://www.cellsignal.com/reference/kinase_disease.htmlhttp://www.cellsignal.com/reference/kinase_disease.html

kinase inhibitors for cancers

Documents