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Kinase 2014: Past, Present and Beyond
New Kinase Inhibitors Targeting theNew Kinase Inhibitors Targeting theNew Kinase Inhibitors Targeting the New Kinase Inhibitors Targeting the Clinical Acquired Resistance Related Clinical Acquired Resistance Related
MutantsMutantsK DiKe Ding
Guangzhou Institutes of Biomedicine and Health, CASg ,
May.19, 2014
(Cambridge UK)
中国科学院广州生物医药与健康研究院 GIBH.CAS
(Cambridge, UK)1
Protein Protein KinasesKinases Are Important Targets for Drug DiscoveryAre Important Targets for Drug Discovery
PProtein kinase
• Cell growth/ proliferation• Differentiation
Dephospho Ph h
P
ATP
kinase• Viability/survival• Homeostasis• Effector function (e gDephospho
proteinsPhosphoproteins
(Modified functions)
Effector function (e.g.cytotoxicity, cytokineproduction)• Cell death
◆About 518 protein kinases were found; an estimated 30,000 genes in humans; about 1.7% of thehuman genome encodes protein kinases
(Modified functions)
human genome encodes protein kinases◆ Protein kinases are the fourth largest gene family in humans:
C2H2 zinc finger proteins (3%)G-protein coupled receptors (2.8%)Major histocompatibility (MHC) complex protein family (2.8%)
◆ Protein kinases are one class of the most popular targets for drug discovery, accounting for 25%
中国科学院广州生物医药与健康研究院 GIBH.CAS2
to 30% of all targets screened in the pharmaceutical industres today.
Kinase Inhibitors Have Achieved Significant Benefits Kinase Inhibitors Have Achieved Significant Benefits for Treatment of Multiple Cancers, However ……for Treatment of Multiple Cancers, However ……p ,p ,
Launched time Generic name Company Targets Indication
2001 Imatinib Novartis Bcr-Abl CMLB Abl
2007 Nilotinib Novartis Bcr-Abl CML
2006 Dasatinib BMS Bcr-Abl CML
Bcr-AblT315I
2002 Gefitinib Astrazenec EGFR NSCLC
2005 Erlotinib OSI Pharm. EGFR NSCLC
2007 L i ib GSK EGFR b i
EGFRT790M
2007 Lapatinib GSK EGFR breast carcinoma
2005 Sorafenib Bayor VEGFR renal carcinoma
2005 Sunitinib Pfizer VEGFR GIST ALK2005 Sunitinib Pfizer VEGFR GIST
2011 Crozotinib Pfizer ALK NSCLC
2011 Icotinib BetaPharm EGFR NSCLC
ALKL1196M
2012 Ponatinib Ariad Pharm Bcr-Abl CML
2013 AfatinibBoehringer
EGFR NSCLC
中国科学院广州生物医药与健康研究院 GIBH.CAS
2013 AfatinibIngelheim
EGFR NSCLC
2013 Ibrutinib Pharmacyclics BtK CLL 3
One of Our Major Research Interests Is: One of Our Major Research Interests Is:
Design and synthesize new kinase inhibitors targeting the clinically acquired
resistance related mutants which may serve as novel therapeutic agentsresistance related mutants, which may serve as novel therapeutic agents.
O O
OHNH2
OHNH2S
Ile 315 Met 318GZD824
Structure information Critical amino acids
Riociguat, Bayer ,
registered,
NNNH2
CO2MeNH2N
NHN
Nregistered,hypertension
N NN
Fand other 60s
N
P i il i t t D lik
中国科学院广州生物医药与健康研究院 GIBH.CAS4
Privileging structures Drug-like core
Recent Research SummaryRecent Research Summary
Di Molecular T f T t D l t St
Recent Research Summary Recent Research Summary
Diseases Targets Types of Target Development Stage
CML Bcr-AblT315I Kinase IND enabling (li d )CML Bcr Abl Kinase (licensed out)
NSCLCs EGFRT790M Kinase Candidate selection
Cancer/ or fibrosis DDR1 Kinase Candidate selection
Melanoma/colon B-RafV600E/EGFR i d i i iMelanoma/colon cancer
B Raf EGFR dual inhibition Kinase Lead optimization
TNBC/metabolic ERR Orphan NHR IND enablingdiseases ERR Orphan NHR (licensed out)
Cancer P53-MDM2 Protein-protein interaction Phase I (Sanofi)
中国科学院广州生物医药与健康研究院 GIBH.CAS5
interaction
Design and synthesis of new Design and synthesis of new BcrBcr--Abl Abl
inhibitors targeting the T315I mutantinhibitors targeting the T315I mutant
中国科学院广州生物医药与健康研究院 GIBH.CAS6
BackgroundBackground::the Unmet Clinical Needsthe Unmet Clinical Needs
• Imatinib achieves significant clinical benefit for CML management . However,
clinically acquired resistance becomes a major challenge (accelerated phase
andblast phase, 50-80%)p
• Bcr-Abl mutation is the primary mechanism for the resistance. More than
100 resistance related Bcr Abl mutants have been identified100 resistance-related Bcr-Abl mutants have been identified.
• The “gatekeeper” T315I is most common mutation. The second-generation
inhibitors (i.e. nilotinib and dasatinib) are not capable of inhibiting Bcr-Abl T315I mutant.
• Bcr-Abl T315I-induced drug resistance remains an unmet clinical challenge
for CML treatment.
中国科学院广州生物医药与健康研究院 GIBH.CAS7
Background: Background: the structure base for molecule designthe structure base for molecule design
• The mechanism for Bcr-Abl T315I mutation induced resistance :h drogen bond loss steric hindrancehydrogen bond loss, steric hindrance.
• Ile315 becomes the most critical residue for new Bcr-AblT315I inhibitord idesign.
ImatinibCh i l F l C H N O
中国科学院广州生物医药与健康研究院 GIBH.CAS8
Thr315 Chemcial Formula: C29H31N7O Ile(315)
Background: Background: the structure base for molecule designthe structure base for molecule design
• Met318 is the 2nd key residue for inhibitor design.
imatinib bosutinib
nilotinib ponatinib
中国科学院广州生物医药与健康研究院 GIBH.CAS9
“Critical Amino Acid” based design of GZD824“Critical Amino Acid” based design of GZD824
GZD824
N NN
O
O H NN
N NN
S
H N
O
N
E tazo la te , B M SP h ase II
P D & A D
N H
C O 2M eNH 2N
G S K -3 56278 , G SKP hase I,
H u ngtington 's d iseasea
R io cig uat B ayer ÁÙ́ ² ½áÊø ´ý Åúh yp ertension
N NN
NNN H 2
中国科学院广州生物医药与健康研究院 GIBH.CAS10
F
Chemical synthesis of GZD824Chemical synthesis of GZD824
中国科学院广州生物医药与健康研究院 GIBH.CAS11
Ren, X.; Ding, K.* J. Med. Chem. 2013, 56 (3), pp 879–894.
GZD824 potently inhibits the T315I mutantGZD824 potently inhibits the T315I mutant
Clinical drugsBiological activities(nM)
Clinical drugsGZD824
imatinib nilotinib dasatinib
Bi di ffi it (K )Abl T315I 1 000 1 000 1 000 0 71Binding affinity(Kd)Abl T315I >1,000 >1,000 >1,000 0.71
Kinase inhibition(IC )
Abl T315I >5,000 >700 >1,400 0.68(IC50)
Abl >5,000 >700 >1,400 0.68
Ba/F3 cell s(IC50) Abl T315I >10,000 >15,000 >3,000 7.1
Normal cells(IC50) HL-7702 >1000 >1000 >1000 >1000
Ren, X.; Ding, K.* J. Med. Chem. 2013, 56 (3), pp 879–894.
中国科学院广州生物医药与健康研究院 GIBH.CAS12
GZD824 also inhibits other resistant mutantsGZD824 also inhibits other resistant mutants
The binding affinities of GZD824 with different Bcr-Abl proteins
c-Abl
Binding affinity (Kd nM)Non-phosphorylated form Phosphorylated form
Wild type 0.32 0.34
T315I 0.71 3.20
Q252H 0.46 NA
E255K NA 0 28E255K NA 0.28
F317I 1.8 NA
H396P 0 18 NA
Ren X ; Ding K * J Med Chem 2013 56 (3) pp 879 894
H396P 0.18 NA
M351T NA 0.23
中国科学院广州生物医药与健康研究院 GIBH.CAS13
Ren, X.; Ding, K.* J. Med. Chem. 2013, 56 (3), pp 879–894.
D824 inhibits growth of cells with resistant mutantsD824 inhibits growth of cells with resistant mutants
Kinase inhibition (IC50,nM)
Bcr-Abl Imatinib Dasatinib GZD824
Wild type 98 2 0 26 0.34Wild type 98.2 0.26 0.34
T315I 5155 1450 0.68
E255K 485.8 0.21 0.27
G250E 359.9 0.25 0.71
Q252H 115.0 0.24 0.15
H396P 173.9 0.30 0.35
M351T 114.3 0.22 0.29
Y253F 749 6 0 17 0 35
中国科学院广州生物医药与健康研究院 GIBH.CAS14Ren, X.; Ding, K.* J. Med. Chem. 2013, 56 (3), pp 879–894.
Y253F 749.6 0.17 0.35
D824 inhibits growth of D824 inhibits growth of BcrBcr--AblAbl positive cellspositive cells
The cell growth inhibitory activities of GZD824 against different cells.organ cell lines GZD824 (X±SD, nM) TAXOL (X±SD, nM)
K-562 0.2±0.1 5.1±0.5K-562R(Q252H)
4.5±0.7 7.0±0.5
BloodKU812 0.1±0.0 5.3±3.2
SUP-B15 2.5±1.0 8.0±3.4
U-937 390.2±153.4 2.3±0.2
MOLT4 26.3±10.2 2.7±0.9
HL-60 348.9±158.2 5.8±1.5
Normal cells HL-7702 1871± 901 4.0±0.6
中国科学院广州生物医药与健康研究院 GIBH.CAS15
Ren, X.; Ding, K.* J. Med. Chem. 2013, 56 (3), pp 879–894.
D824 demonstrates promising D824 demonstrates promising in vivo in vivo efficacyefficacy
中国科学院广州生物医药与健康研究院 GIBH.CAS16
Ren, X.; Ding, K.* J. Med. Chem. 2013, 56 (3), pp 879–894.
D824 demonstrates promising D824 demonstrates promising in vivo in vivo efficacyefficacy
GZD824 efficiently prolongs animal survival in an allograft tumor model using Ba/F3 cells co expressing Bcr Abl T315I and luciferase
Day 0Ba/F3 cells co-expressing Bcr-Abl T315I and luciferase.
Day 7
100 mg/kgImatinib
Vehicle 1 mg/kg 10 mg/kg 20mg/kg5 mg/kg2 mg/kg
中国科学院广州生物医药与健康研究院 GIBH.CAS
Imatinib GZD82417
D824 displays good drugD824 displays good drug--like propertieslike properties
The drug-like properties of GZD824:g p p
1)PK profile: F = 48.7%;T1/2 = 8-10 hrs;no inhibition against
CYPs at high concentrations (over 1000 folds);
2)S f H G IC 50 M f i d 10 f ld2)Safety: HerG,IC50 > 50 M;safety window > 10 folds;
3)Currently is under IND enabling investigation (Guangzhou3)Currently is under IND enabling investigation (Guangzhou
Dunjian Pharm.)1. Ren, X.; Ding, K.* etc. J. Med. Chem. 2013, 56 (3), pp 879–894
2. Li, Y.; Ding, K.* etc. J. Med. Chem. 2012, 55 (22), 10033-10046.
3. Ke Ding, Deping Wang, Duanqing Pei, Zhang Zhang, etc. Chinese patent granted, # ZL 201010216603.7 ;
中国科学院广州生物医药与健康研究院 GIBH.CAS18
PCT application:WO 2012/000304
Discovery of Novel Selective EGFRT790M
Mutant Inhibitors Overcoming the
Clinically Acquired Resistance against
Gefinitib
中国科学院广州生物医药与健康研究院 GIBH.CAS19
BackgroundBackground::EGFR Inhibitor & NSCLCEGFR Inhibitor & NSCLC• EGFR is a well validated target for anticancer drug discovery。
• 2003,FDA approved Gefitinib (Iressa) for the treatment of NSCLC patients with2003,FDA approved Gefitinib (Iressa) for the treatment of NSCLC patients with
EGFR activating mutation (L858R, del E746_A750). The first exapmle for
personalized medicine.p
• 2004,Erlotinib was approved by FDA .
• 2011 Icotinib was approved by SFDA• 2011,Icotinib was approved by SFDA.
• 2013,the irresible 2nd inhibitor Gilotrif (BIBW-2992) was approved
EGFR mutation)NSCLC
patients
(L858R,del E746_A750)
EGFR wild type Other
中国科学院广州生物医药与健康研究院 GIBH.CAS20
(or other mutations) therapies
EGFREGFRT790M T790M Mutation & Acquired ResistanceMutation & Acquired Resistance
• The first generation inhibitors, gefinitib and erlotinib have achievedsignificant clinical benefits However emerging acquired resistance to themsignificant clinical benefits. However, emerging acquired resistance to themhas become a major clinical challenge.
• EGFRT790M mutation is the primary mechanism for the resistance against• EGFR mutation is the primary mechanism for the resistance againstIressa in NSCLC patients(50% of resistant NSCLC patients)
• EGFRT790M mutation only moderately affects the binding of gefinitib andEGFR mutation only moderately affects the binding of gefinitib anderotinib, but significantly enhances the binding affinity for ATP with EGFR.
EGFRWT
Km = 5.2 MEGFRL858R
Km = 148 MEGFRL858R/T790M
Km = 8.4 M
Gefitinib Insensitive Gefitinib Sensitive Gefitinib Resistant
中国科学院广州生物医药与健康研究院 GIBH.CAS21
EGFREGFRT790M T790M Mutation & Acquired ResistanceMutation & Acquired Resistance
• The 2nd generation irreversible inhibitors: effective in animal models, butlow MTD in clinical investigation; dose-limit toxicity. The drugconcentrations are not high enough to inhibit EGFRT790M at MTD poorconcentrations are not high enough to inhibit EGFRT790M at MTD, poorefficacy. Most of the clinical trials are halted.
• Key problem: no selectivity over EGFR WT.ey p ob e : o se ect v ty ove G .
SHN
HNHN Cl
F
O
CNHN
HN Cl
N
ON
--SH NONO
Canertinib
NOO
N
Neratinib
FHN Cl
F
N
N
O
HN
N
HN Cl
ON
NHN
O
HN Cl
ON
O
• EGFRT790M induced drug resistance remains an unmet!!
DacomitinibO
Afatinib
中国科学院广州生物医药与健康研究院 GIBH.CAS
clinical challenge for NSCLCs treatment!!22
Selective EGFRSelective EGFRT790MT790M Inhibitors Overcoming ResistanceInhibitors Overcoming Resistance
• Inhibitors selectively targeting EGFRT790M mutants become a novel attractive strategy
for clinical management of NSCLC patients with acquired resistance. However:g p q
• EGFRWT and the EGFRT790M mutants share highly similar 3-dimensional structures and
have almost identical binding affinities with ATP.have almost identical binding affinities with ATP.
• The development of WZ4002 is halted due to IP issue; CO-1686 and AZD9291 are
currently in phase I/II trial But the selectivity is relatively low (25-40 folds)currently in phase I/II trial. But the selectivity is relatively low (25-40 folds).
中国科学院广州生物医药与健康研究院 GIBH.CAS
W. Zhou et al. Nature 462(2009) 1-70-107423
“Critical Amino Acid” based design of EGFR “Critical Amino Acid” based design of EGFR T790MT790M
inhibitorsinhibitorsinhibitorsinhibitors
Met793Met790Met790
D101
Xu T ; Ding, K.* Angew. Chem. Int. Edt. 2013 DOI: 10 1002/anie 201302313.
中国科学院广州生物医药与健康研究院 GIBH.CAS
Xu, T.; Ding, K. Angew. Chem. Int. Edt. 2013, DOI: 10.1002/anie.201302313. 24
The compound displays great selectivity on EGFR The compound displays great selectivity on EGFR T790MT790M
mutant over the wildmutant over the wild--typetype kinasekinaseBinding affinities(Kd):
EGFR WT 310 M
mutant over the wildmutant over the wild type type kinasekinase
EGFR WT = 310 nM
EGFR T790M = 1.3 nM
EGFR L858R/T790M = 2.3 nM
Kinase inhibition IC50):
EGFR T790M = 4.55 nM
EGFR L858R/T790M = 2.18 nM
Selectivity(120-240 folds):
Kd (EGFRWT)/ Kd (EGFR L858R/T790M ) = 119.2d ( ) d ( )
Kd (EGFRWT)/ Kd (EGFRT790M ) = 238.5
No obvious inhibition against 455 different kinases
中国科学院广州生物医药与健康研究院 GIBH.CAS
No obvious inhibition against 455 different kinasesevaluated as 100nM.
25
The compound displays great selectivity on EGFR The compound displays great selectivity on EGFR T790MT790M
mutant over the wildmutant over the wild typetype kinasekinase
Cells lines EGFR status Anti-proliferation (IC50, M)
mutant over the wildmutant over the wild--type type kinasekinase
p 50
H1975 L858R/T790M 0.086±0.018H332 WT >30A549 WT >30
H1299 WT >30H1703 WT >30H661 WT >3095D WT >3095D WT >30
H358 WT >30
HCC827 del E746 A750 0.049±0.027_
HL-7702 WT >30HLF-1 WT 10.50±1.46
中国科学院广州生物医药与健康研究院 GIBH.CAS
The most selective EGFRT790M inhibitor reported to date. 26
The compound potently inhibit the activation of The compound potently inhibit the activation of EGFR signal pathwayEGFR signal pathway
0 0 016 0 08 0 4 2 0
3d
0 0 01 0 1
Gfb
0 0 016 0 08 0 4 2 0
3g
0 0.016 0.08 0.4 2.0 0 0.01 0.1
EGFR
0 0.016 0.08 0.4 2.0
pEGFR
Akt
pAkt
Erk
pErkpErk
GAPDH
中国科学院广州生物医药与健康研究院 GIBH.CAS27
The compound doseThe compound dose--dependently dependently inhibit the inhibit the migration and invasion of NCImigration and invasion of NCI--1975 NSCLC cells1975 NSCLC cellsgg
中国科学院广州生物医药与健康研究院 GIBH.CAS28
The compound doseThe compound dose--dependently dependently inhibit the colony inhibit the colony formation of NCIformation of NCI--1975 cells1975 cells
中国科学院广州生物医药与健康研究院 GIBH.CAS29
New compounds are obtained with improved PK New compounds are obtained with improved PK profilesprofiles
A D Animal Dose AUC (0-∞) C
profilesprofiles
Cpd. A.D.route
Animal No. level
mg/kg
AUC (0-∞) mg/L*h T1/2 (h) Tmax (h) Cmax
g/L F(%)
PO ♂4 25 1640 1.695 0.88 10405-1 30.0
IV ♂4 5 2491 0.617 0.03 3975
1. Xu, T.; and Ding, K.* Angew. Chem. Int. Edt. 2013, 52, 8387–8390.
2. Chang, S.; Ding, K. * J. Med. Chem. 2012, 55 (6), 2711-2723.
3. Han, C.; Ding, K.*; Ji, H.*; Tian, J. and Zhang, Y.* J. Med. Chem. 2013, 56, 4738–4748.
4. Xu, S.; Ding, K.* MedChemComm. 2012, 3, 1155-1159.
5. Xu. S. ; and Ding, K.* J. Med. Chem. 2013, revision.
中国科学院广州生物医药与健康研究院 GIBH.CAS30
The new compounds shows good The new compounds shows good in vivo in vivo efficacyefficacy
中国科学院广州生物医药与健康研究院 GIBH.CAS31
SummarySummary
Starting From the Unmet Clinical Needs:
Obtained the Bcr-Abl T315I inhibitor GZD824 as new
candidate to manage the clinically acquired resistance
against Imatinib;g ;
Designed and synthesized highly selective EGFRT790M
inhibitors which may potentially used to manage the
clinically acquired resistance against Gefitinibclinically acquired resistance against Gefitinib .
中国科学院广州生物医药与健康研究院 GIBH.CAS32
Acknowledgements Acknowledgements • Collaborators
1)Biologists/Pharmacologists:SIMM、SYSU、GIBH etcSIMM、SYSU、GIBH etc.
2) Medicinal Chemistry:Graduate
students and Ras.
• Financial SupportFinancial SupportNSFC, Ministry of S & T, CAS,
Guangdong province and Guangzhou
city.
中国科学院广州生物医药与健康研究院 GIBH.CAS33
Thank You!
中国科学院广州生物医药与健康研究院 GIBH.CAS34