Upload
dinhtu
View
236
Download
3
Embed Size (px)
Citation preview
The Novel IL-2 Cytokine Immune Agonist NKTR-214 Harnesses the Adaptive and Innate Immune System for the Treatment of Solid CancersSalah Eddine Bentebibel1, Chantale Bernatchez1, Cara Haymaker1, Michael Hurwitz3, Patrick Hwu1, Mario Sznol3, Nizar Tannir1, Anthony Conley1, Harriet Kluger3, Sandra Aung2, Michael Imperiale2, Mary Tagliaferri2, Christie Fanton2, Ernesto Iacucci2, Jonathan Zalevsky2, Ute Hoch2, Adi Diab1
1The University of Texas MD Anderson Cancer Center, Houston, TX, United States of America, 2Nektar Therapeutics, San Francisco, CA, United States of America, 3Yale School of Medicine, New Haven, CT, United States of America
BACKGROUND
RESULTS
RESULTS
NKTR-214• NKTR-214 is a CD122-biased cytokine agonist
conjugated with multiple releasable chains of polyethylene glycol (PEG) designed to provide sustained signaling through the heterodimeric IL-2 receptor pathway (IL-2R) to preferentially activate and expand effector CD8+ T and NK cells over Tregs1 (Figure 1).
NKTR-214 MONOTHERAPY STUDY• A phase 1, multicenter, open-label, dose-
escalation study (EXCEL) was conductedto assess the safety, preliminary efficacy, pharmacokinetics, and pharmacodynamicsof NKTR-214 in 28 patients with locally advanced or metastatic solid tumors.2-4
• Outpatient regimen with a convenient IV dosing regimen every 2 or 3 weeks.
• NKTR-214 has a favorable safety and tolerability profile.4
– No evidence of immune-mediated AEs or organ related inflammation (eg, colitis, pneumonitis, dermatitis, hepatitis, endocrinopathies). – Grade 3 hypotension occurred in 14% of
patients and was rapidly reversible with IV fluids. – No patients experienced capillary leak syndrome. – No Grade 4 TRAEs or treatment-related deaths
• Maximum-administered dose (MAD) was 0.012 mg/kg q3w.
• Sustained exposure and robust PD changes after a single dose of NKTR-214 (Figure 3).3,5
• NKTR-214 substantially increased CD8+ T cells that were newly proliferative (Ki-67+) (Figure 4).5
CONCLUSIONS
Figure 1. NKTR-214 Mechanism of Action
CLONAL EXPANSION
Stimulates Immune Response to Kill Tumor Cells
LEGEND:NKTR-214 – Inactive2-PEG – Active Cytokine1-PEG – Active Cytokine
NKTR-214 (6-PEG)
IrreversibleRelease
2-PEGActive Cytokine
1-PEG Active Cytokine
IrreversibleRelease
IL-2Rαβγ
α
β γβ γ
IL-2Rβγ
Immunosuppressivecells limit anti-tumor
responseNKNK
CD8+
CD8+
CD8+
CD4+
Helper
CD4+
Helper
CD4+
Treg
NK NK
NK, CD4+, and CD8+ T cells
CD4+
HelperCD8+
CD4+
Helper
CD4+
HelperCD8+
NK CD4+
Helper
NK
CD8+
CD4+
HelperCD4+
HelperCD8+
CD8+
NKNK
NK
CD8+
CD4+
Helper
CD4+
Helper
NKCD8+
REFERENCES1) Charych DH, Hoch U, Langowski JL, et al. Clin Cancer Res. 2016;22(3):680–90.2) Bernatchez C, Haymaker C, Tannir NM, et al. Presented at SITC 2016, National Harbor, MD. Poster #387.3) Hurwitz ME, Diab A, Bernatchez C, et al. Presented at ASCO GU 2017, Orlando, FL. Poster #D17.4) Bernatchez C, Bentebibel SE, Hurwitz ME, et al. Presented at ASCO 2017, Chicago, IL.5) Nektar Therapeutics Analyst & Investor Event at ASCO 2017 Annual Meeting. [online]
Available at: http://ir.nektar.com/events.cfm
Figure 2. Time on Study and Best Overall ResponseSD
SDSD
SDSD(uPR)
SDSD
PDSD
PDPD
PDPDPD
PDPD
SDSD
SDSD
SDNE
PDSD
PDSDPD
NE
0.003 mg/kg q3w0.006 mg/kg q3w0.012 mg/kg q3w0.009 mg/kg q3w0.006 mg/kg q2w
Discontinued treatment due to RECIST PDDiscontinued treatment due to AEDiscontinued treatment due to other reasons
PD - Best Overall Response is Progressive DiseaseSD - Best Overall Response is Stable Disease
Data cut-off: October 4, 2017
W3/C1 W15/C5 W30/C10 W45/C15 W60/C20 W75/C25 W90/C30
Breast CaRCCRCC
ChondrosarcomaLeiomyosarcoma
RCC
MelanomaRCCRCC
MelanomaRCCRCCRCC
Triple Neg BCMelanomaMelanoma
RCCColorectal
RCCRCCRCCRCCRCCRCC
MelanomaBladder CaMelanomaMelanoma
Time on Study (Weeks/Cycles)0 W8/C4 W16/C8 W24/C12 W32/C16 W40/C20 W48/C24 W56/C28 W64/C32 W72/C36 W80/C40 W88/C44 W96/C48 W104/C52
Table 1. NKTR-214 induces gene expression changes in tumors associated with innate and adaptive immune pathways
• NanoString analysis includes 10 patients with matched baseline and wk3 samples and 5 additional patients with either baseline or wk3 sample
− RCC (8), melanoma (4), chondrosarcoma (1), leiomyosarcoma (1), and triple negative breast cancer (1)• 89 genes differentially expressed between pre- and post-treatment when all patients used in the analysis
Pathway name (database source) pSize Genes in pathway differentially expressed post-treatment
T cell receptor signaling pathway (KEGG) 37 ICOS; ITK; LCK; MAPK11; ZAP70; CD3E; CD247; CD8A; CD8B; CD40LG; CTLA-4; PDCD1
TCR signaling in naïve CD8+ T cells (NCI) 20 CD8A; CD3D; LCK; CD3E; CD3G; CD8B; CD247; ZAP70; PRF1
IL-12-mediated signaling events (NCI) 43 SOCS1; IFNG; IL-2RA; CD8A; CD3D; CD3E; CD3G; CD8B; IL-2RB; CD247; IL-2RG; STAT4; IL-12RB2; EOMES; LCK; GZMB; TBX21
IL-2-mediated signaling events (NCI) 21 SOCS1; IL-2RA; LCK; IL-2RB; IL-2RG; MAPK11; IFNG
Nature killer cell mediated cytotoxicity (KEGG) 48 GZMB; IFNG; KLRC1; KLRC2; KLRD1; LCK; SH2D1A; CD244;ZAP70; CD247; KLRK1; NCR1; BID; SH2D1B; TNFRSF10C
pSize: Total # of genes in pathway covered by NanoString assay
Figure 11. Case study: Patient with RCC stage IV, age 59, male, progressed on prior TKI
TreatmentDuration ofTreatment
Time Interval toNext Treatment
Prior Therapy Sunitinib ~ 67 mos (PD) ~ 2 mos
TherapiesAdministered
NKTR-214 ~ 5 mos (SD) ~ 1 mo
OngoingNivolumab > 9 mos
H&E CD3 CD8
Baseline
Week 3
Immunological Changes onNKTR-214 Therapy
1027 meancells/mm2
675 meancells/mm2
2017 meancells/mm2
1620 meancells/mm2
EOT NKTR-214 POST Nivolumab
~1 mo
Con�rmed PRPR on 1st scan
Nivolumab
Pre NKTR-214Left Lung Nodule
NKTR-214 (8 cycles)PD-L1 status† = neg
(Max tumor response -10% per RECIST 1.1)
†PD-L1 status was obtained using the Cell Signaling antibody (Cell Signaling #13684, PD-L1 (E1L3N)) at a 1:100 dilution on the Leica BOND RXm
Robust immune-stimulatory responseIncrease in T cell receptor
clonality at wk3
• Increased T cell clonality and induction of immune-related genes in the tumor, along with an increase in the proliferative index of immune cells in the blood, indicate the ability of NKTR-214 to induce robust immune changes in patients
1.00
0.75
0.50
0.25
0.000.00 0.25 0.50
Cumulative percentage ofunique sequences
Cum
ulat
ive
per
cent
age
of
read
s
0.75 1.00
BaselineWeek 3
Increase in proliferationand PD-1+ expressionfollowing NKTR-214
0
10
20
30
40
0
10
20
30
40
50
Day
CD
4+ K
i-67
+ (%
CD
4)
CD
8+ K
i-67+ (%
CD
8)
C1D1 C1D8
CD4+ Ki67+
CD8+ Ki67+
CD
4+ K
i-67
+ P
D-1
+ (%
CD
4)
CD
8+ K
i-67+ P
D-1
+ (%C
D8)0
2
4
6
0
5
10
15
Day
CD4+ Ki67+ PD-1+
CD8+ Ki67+ PD-1+
C1D1 C1D8
>2-fold induction ofimmune checkpoint genes and
CD8-related genes at wk3
0 2 4 6 8 10Fold change (wk3/baseline)
LAG3ICOSIDO1
CTLA-4TIGITPD-1
PD-L1PD-L2
Tim3OX40TP53CD19
0 1 2 3 4 5 6 7Fold change (wk3/baseline)
IFNGPRF1
GZMBTBX21CD8ACD8B
IL-6SMAD3
• Sustained exposure with half-life of ~20 hours• Gradual increase in active cytokine species,
reaching Cmax 1-2 days post dose• Exposure increases in proportion to dose• Free IL-2 is not detectable
• Transient decrease followed by increase in lymphocytes from Day 3 to Day 9• Transient increase in soluble IL-2 receptor alpha (sCD25) from Day 1 to Day 8,
shed from activated T cells• sCD25 levels return to baseline by Day 15• Similar PD effects observed across dose levels
Pharmacodynamics(0.006 mg/kg, n=15)
Figure 3. Sustained exposure and robust PD changes after a single dose of NKTR-214
RC: Related Cytokine, total protein detection assay measures 6-PEG NKTR-214 and all derived species; AC: Active Cytokine, detection assay for 2-PEG and lower active species
Lymphocytes
0
1
2
3
109 /
L (M
ean+
SE
)
Time (Days)15 221 8
Time (Days)
sCD25
15 221 8
0
2
4
6
ng/m
L (M
ean+
SE
)
Pharmacokinetics(0.006 mg/kg, n=15)
ng/m
L (M
ean+
SE
)
NKTR-214-AC
NKTR-214-RC
15 221 8Time (days)
0.1
1
10
100
1000
Tumor Analysis Fresh TIL analysis by �ow cytometry IHC T cell receptor gene sequencing Gene expression analysis
Blood Analysis Flow cytometry Cytokines PK PD (sCD25, lymphocytes)
Blood and Tumor Biopsy Collection and Analysis
EOT, end of treatment.
C1D1Cycle (C)/Day (D): C1D8 C2D1
Q2W or Q3W
C3D1
EOT
NKTR-214
C2D8
BaselineTumorBiopsies: Week 3
= Blood Sample
Week 8 EOT
NKTR-214NKTR-214
ACKNOWLEDGEMENTS• The authors would like to acknowledge the contribution of patients and their families in
participation of this clinical trial.
Figure 4. NKTR-214 promotes proliferation of CD4+ T cells, CD8+ T cells, and NK cells in peripheral blood
CD4+ T cells
DayC1D1
n=16
C1D80
20
40
60
10
30
50
70
CD
4+ K
i-67
+ (%
CD
4)
Melanoma
ChondrosarcomaBreast CancerBladder Cancer
Renal Cell Carcinoma
Ki-67+ set on baseline sample at ~5%
• An increase in overall T cell clonality observed in 8/12 patients after NKTR-214 treatment
• Increase in clonality associated with increased T cell infiltrate in 7/12 patients
Figure 10. Increase in T cell clonality in tumors after treatment with NKTR-214
Melanoma
ChondrosarcomaRenal Cell Carcinoma
-6 -5 -4
-4-3-2-1012345
0 21 3 4Clonality (Log2)
In�
ltra
te (L
og
2)
Figure 5. NKTR-214 induces activating markers and co-inhibitory receptors in proliferating CD4+ T cells in peripheral blood
Ki-
67+
Ki-
67−
0
20
40
60
80
Day
CD
4+ K
i67+
ICO
S+ (%
CD
4)
C1D1 C1D8
n=15
PD-1 CTLA-4 OX40 ICOS
DayDay
0
5
10
15
CD
4+ K
i-67
− P
D-1
+ (%
CD
4)
C1D1
n=10
C1D8Day
0
5
1020
30
40
C1D1 C1D8
n=16
CD
4+ K
i-67
− C
TLA
-4+ (%
CD
4)
n=8
0
5
10
15
20
C1D1 C1D8
CD
4+ K
i-67
− O
X40
+ (%
CD
4)
n=9
0
5
10
15
20
DayC1D1 C1D8
CD
4+ K
i-67
+ O
X40
+ (%
CD
4)
0
5
1020
30
40
DayC1D1 C1D8
n=15
CD
4+ K
i-67
+ C
TLA
-4+ (%
CD
4)
0
5
10
15
DayC1D1
n=11
C1D8
CD
4+ K
i-67
+ P
D-1
+ (%
CD
4)
Activating and co-inhibitory marker gates set on C1D1 sample Legend is same as for Figure 6
0
20
40
60
80
C1D1 C1D8
n=15
Day
CD
4+ K
i-67
− IC
OS
+ (%
CD
4)
ICOS
Figure 6. NKTR-214 induces activating markers and co-inhibitory receptors in proliferating CD8+ T cells in peripheral blood
0
2
48
10
C1D1 C1D80
2
4
6
8
C1D1 C1D80
5
10
15
20
25C
D8+
Ki-
67− P
D-1
+ (%
CD
8)C
D8+
Ki-
67+ P
D-1
+ (%
CD
8)
CD
8+ K
i-67
+ C
TLA
-4+ (%
CD
8)C
D8+
Ki-
67− C
TLA
-4+ (%
CD
8)
CD
8+ K
i-67
− IC
OS
+ (%
CD
8)C
D8+
Ki-
67+ IC
OS
+ (%
CD
8)
C1D1 C1D8
0
2
48
10
C1D1 C1D80
2
4
6
8
C1D1 C1D80
5
10
15
20
25
Day Day Day
Day Day Day
C1D1
n=8n=12n=8
n=10n=11n=10
C1D8K
i-67
+K
i-67
−
PD-1 CTLA-4
Activating and co-inhibitory marker gates set on C1D1 sample
Melanoma
ChondrosarcomaBreast CancerBladder Cancer
Renal Cell Carcinoma
CD8+ T cells
DayC1D1 C1D8
CD
8+ K
i-67
+ (%
CD
8)
0
20
40
60
10
30
50
70n=16
CD3-CD56+ NK cells
DayC1D1 C1D8
n=13
CD3- C
D56+ K
i-67+ (
%CD
56)
0
20
40
60
80
100
Case Study
• Immunological activity in peripheral blood and tumor tissue consistent with NKTR-214’s biological mechanism of biased IL-2 pathway activation
• NKTR-214 has a robust PK-PD profile
• Robust immune-stimulatory response in the tumor and blood– NKTR-214 induced change in proliferative index of immune cells and
costimulatory/checkpoint markers only on newly proliferating immune cells– RNA expression changes consistent with increased immune cell infiltrate and
activation of effector mechanisms– Increased frequency of specific T cell clones and associated increase in T cell
infiltrate suggests efficient remodeling of the T cell repertoire
• The ability of NKTR-214 to increase TILs and increase PD-1 expression on immune cells provides a sound biological basis for combination with anti-PD1 checkpoint inhibitors
• Clinical trials currently enrolling patients with NKTR-214 in combination with anti-PD1 checkpoint inhibitors:
– PIVOT-02 clinical trial combining NKTR-214 + nivolumab in patients with metastatic melanoma, renal cell carcinoma, non-small cell lung cancer, urothelial cancer and triple-negative breast cancers (NCT02983045)
– PROPEL clinical trial combining NKTR-214 with atezolizumab or pembrolizumab in patients with metastatic melanoma, urothelial and non-small cell lung cancers (NCT03138889)
– A study of NKTR-214 + nivolumab in patients with metastatic and/or locally advanced sarcoma (NCT03282344)
Each line represents a different patient
Each line represents a different patient
Each line represents a different patient
Poster #P77: Society for Immunotherapy of Cancer 2017 Annual Meeting
Figure 9. NKTR-214 does not alter expression of Th17- or Treg-associated genes in the tumor
Gene nameGenesymbolRORC ROR gamma; transcription factor
TGFB1 TGF beta 1
TGFB2 TGF beta 2
IL-10 Interleukin-10
0.0
5.0
10.0
15.0
2.5
7.5
12.5
TGFB2
Log
−co
un
ts
RORC TGFB1 IL-10
Figure 8. NKTR-214 induces immune checkpoint and co-inhibitory genes in the tumor
Genesymbol Gene name
PDCD1 PD-1
CTLA-4 Cytotoxic T lymphocyte associated protein 4
LAG3 Lymphocyte activating 3
ICOS Inducible T cell costimulator
IDO1 Indoleamine 2,3-dioxygenase 1
TIGIT T cell immunoreceptorwith Ig and ITIM domains
MeanMedian
Figure 7. NKTR-214 induces gene expression changes in the tumor associated with T cell activation
Genesymbol Gene name
CD8A CD8 alpha chain isoform
CD8B CD8 beta chain isoform
TBX21 Th1 cell-specific transcription factor
IFNG Interferon gamma
GZMB Granzyme B
PRF1 Perforin-1
p-value ≤0.05 for all genes, comparing baseline vs wk3 means
p-value ≤0.05 for all genes, comparing baseline vs wk3 means
p-value ≥ 0.1 for all genes, comparing baseline vs wk3 means
Baseline n=14Week 3 n=11
MeanMedian
Baseline n=14Week 3 n=11
MeanMedian
Baseline n=14Week 3 n=11
0.0
2.5
5.0
7.5
10.0
12.5
CD8A CD8B TBX21 IFNG GZMB PRF1
Lo
g−
cou
nts
0.0
4.0
8.0
2.0
6.0
10.0
12.0
PDCD1 CTLA-4 LAG3 ICOS IDO1 TIGIT
Log
−co
un
ts