Lytix Biopharma AS | Gaustadalléen 21, NO-0349 Oslo, Norway | E-mail: [email protected]| Phone: +47 77 67 55 00 | Fax: +47 77 67 55 01 LTX-315: A first-in-class oncolytic peptide that reshapes the tumor microenvironment BALDUR SVEINBJØRNSSON 2,3 , KETIL ANDRÉ CAMILIO 1,2 , MENG-YU WANG 1 , JANNE NESTVOLD 1,2 , AND ØYSTEIN REKDAL 2,3 1. INSTITUTE OF CANCER RESEARCH, OUS, OSLO NORWAY 2. LYTIX BIOPHARMA AS, P.O. BOX 6447, NO-9294 TROMSØ, NORWAY 3. DEPARTMENT OF MEDICAL BIOLOGY, FACULTY OF HEALTH SCIENCES, UNIVERSITY OF TROMSØ, TROMSØ, NORWAY Background The oncolytic peptide LTX-315, which has been de novo designed based on structure-activity relationship studies of host-defense peptides, has the ability to kill human cancer cells and induce long-lasting anticancer immune response when injected locally into tumors established in immunocompetent murine models (1-3,11). The oncolytic effect of LTX-315 involves perturbation of the plasma membrane and the mitochondria with subsequent release of danger- associated molecular pattern molecules (DAMPs) such as ATP, Cytochrome C and HMGB1 (4-9). Furthermore, LTX-315 effectively disintegrates the cellular compartments with subsequent release of tumor antigens as demonstrated by a greater T-cell infiltration (TILs), TILs clonality and the number of clones with greater abundance in the tumor microenvironment. In experimental tumor models, LTX-315 exerts abscopal effects and reshapes the tumor microenvironment by decreasing the local abundance of immunosuppressive cells and by increasing the frequency of effector T-cells (9,10). LTX-315´s ability to convert immunogenically “cold” tumors to “hot” makes it ideal combination partner with other immunotherapies as confirmed in experimental tumors combining LTX-315 with immune checkpoint inhibitors and immunochemotherapy. LTX-315 Mode of action LTX-315 induces a unique type of immunogenic cell death WITH CHECKPOINT INHIBITORS WITH CHEMOTHERAPY Anti PD-1 Control tumor LTX-315 increases the number and diversity of T cell clones T cell clones in LTX-315-treated and control tumors (B-16 melanoma) were amplified and sequenced using the ImmunoSeq platform by Adaptive Biotech. Multiplex PCR was used to amplify the rearranged TCR3b sequences from sample DNA (VDJ region). Expanded Stable Contracted LTX-315 treated tumor Anti CTLA-4 Low dose cyclophosphamide (A20 B-lymphoma) Doxorubicin (4T1 breast cancer) Conclusion • LTX-315 shows an enhanced anticancer efficacy against A20 lymphomas and 4T1 breast carcinomas when combined with cyclophosphamide and doxorubicin, respectively. • LTX-315 acts in synergy with checkpoint inhibition. • The LTX-315 unique “release and reshape” properties make it a promising candidate for combination with several types of immunotherapies. • LTX-315 is currently in clinical phase 1/2a studies. References 1. Haug et al. J Med Chem. 2016 2. Camilio et al. Cancer Immunol Immunother. 2014 3. Camilio et al. Oncoimmunology. 2014 4. Zhou et al. Oncotarget. 2015 5. Eike et al. Oncotarget. 2015 6. Forveille et al. Cell Cycle. 2015 7. Zhou et al. Cell Death Dis. 2016 8. Sistigu et al. Cell Cycle. 2016 9. Yamazaki et al. Cell Death Differ. 2016 10. Nestvold et al. Oncoimmunology. 2017 11. Sveinbjørnsson et al. Future Medicinal Chemistry. 2017 LTX-315 treatment leads to increased tumor infiltration of CD8+ T cells LTX-315 demonstrates synergy with other standard of care cancer therapies Treatment induces T-cell infiltration into both treated primary lesions but also in distal non-treated tumors (abscopal effect) LTX-315 CD3+ CD8+ Untreated Untreated Treated Treated Tumor Control 0 10 20 30 0 100 200 300 400 days after treatment days days post tumor challenge Tumor size (mm 2 ) % Survival Tumor size (mm 2 ) Control LTX-315 aCTLA-4 aCTLA-4 + LTX-315 Controls (0/8) Cyclophosphamide (0/7) LTX-315 (1/8) LTX-315 + cyclophosphamide (6/9) days post tumor challenge % Survival Controls (0/9) Doxorubicin (0/9) LTX-315 (0/8) LTX-315 + doxorubicin (5/10) Iso Ctrl + PBS Iso Ctrl + LTX-315 aPD1 + PBS aPD1 + LTX-315 N H O NH 2 N H O N H O N N H O NH 2 N H O NH 2 N H O NH N H O N H 2 O N H O NH NH 2 NH 2 NH 2 OH O n
LTX-315: A first-in-class oncolytic peptide that reshapes the tumor microenvironment
BALDUR SVEINBJØRNSSON2,3, KETIL ANDRÉ CAMILIO1,2, MENG-YU WANG1, JANNE NESTVOLD1,2, AND ØYSTEIN REKDAL2,3
1. INSTITUTE OF CANCER RESEARCH, OUS, OSLO NORWAY2. LYTIX BIOPHARMA AS, P.O. BOX 6447, NO-9294 TROMSØ, NORWAY3. DEPARTMENT OF MEDICAL BIOLOGY, FACULTY OF HEALTH SCIENCES, UNIVERSITY OF TROMSØ, TROMSØ, NORWAY
BackgroundThe oncolytic peptide LTX-315, which has been de novo designed based on structure-activity relationship studies of host-defense peptides, has the ability to kill human cancer cells and induce long-lasting anticancer immune response when injected locally into tumors established in immunocompetent murine models (1-3,11).
The oncolytic effect of LTX-315 involves perturbation of the plasma membrane and the mitochondria with subsequent release of danger-associated molecular pattern molecules (DAMPs) such as ATP, Cytochrome C and HMGB1 (4-9). Furthermore, LTX-315 effectively disintegrates the cellular compartments with subsequent release of tumor antigens as demonstrated by a greater T-cell infiltration (TILs), TILs clonality and the number of clones with greater abundance in the tumor microenvironment.
In experimental tumor models, LTX-315 exerts abscopal effects and reshapes the tumor microenvironment by decreasing the local abundance of immunosuppressive cells and by increasing the frequency of effector T-cells (9,10). LTX-315´s ability to convert immunogenically “cold” tumors to “hot” makes it ideal combination partner with other immunotherapies as confirmed in experimental tumors combining LTX-315 with immune checkpoint inhibitors and immunochemotherapy.
LTX-315
Mode of action
LTX-315 induces a unique type of immunogenic cell death
WITH CHECKPOINT INHIBITORS
WITH CHEMOTHERAPYMode of action
Anti PD-1
Control tumor
LTX-315 increases the number and diversity of T cell clones
T cell clones in LTX-315-treated and control tumors (B-16 melanoma) were amplified and sequenced using the ImmunoSeq platform by Adaptive Biotech. Multiplex PCR was used to amplify the rearranged TCR3b sequences from sample DNA (VDJ region).
LTX-315 increases T-cell clonality
LTX-315treatedtumor Controltumor
Expanded
Stable
Contracted
T cell clones in LTX-315-treated and control tumors were amplified and sequenced using the ImmunoSeq platform by Adaptive Biotech. Multiplex PCR was used to amplify the rearranged TCR3b sequences from sample DNA (VDJ region).
LTX-315 treated tumor
Anti CTLA-4
Low dose cyclophosphamide (A20 B-lymphoma)
Doxorubicin(4T1 breast cancer)
Conclusion• LTX-315 shows an enhanced anticancer efficacy against A20 lymphomas
and 4T1 breast carcinomas when combined with cyclophosphamide and doxorubicin, respectively.
• LTX-315 acts in synergy with checkpoint inhibition.
• The LTX-315 unique “release and reshape” properties make it a promising candidate for combination with several types of immunotherapies.
• LTX-315 is currently in clinical phase 1/2a studies.
References1. Haug et al. J Med Chem. 20162. Camilio et al. Cancer Immunol Immunother. 20143. Camilio et al. Oncoimmunology. 2014 4. Zhou et al. Oncotarget. 2015 5. Eike et al. Oncotarget. 2015 6. Forveille et al. Cell Cycle. 20157. Zhou et al. Cell Death Dis. 2016 8. Sistigu et al. Cell Cycle. 20169. Yamazaki et al. Cell Death Differ. 201610. Nestvold et al. Oncoimmunology. 201711. Sveinbjørnsson et al. Future Medicinal Chemistry. 2017
LTX-315 treatment leads to increased tumor infiltration of CD8+ T cells
LTX-315 demonstrates synergy with other standard of care cancer therapies
Treatment induces T-cell infiltration into both treated primary lesions but also in distal non-treated tumors (abscopal effect)
LTX-315
CD3+ CD8+
Untreated UntreatedTreated TreatedTumor
Control
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
days after treatmentdays
days post tumor challenge
Tum
or s
ize
(mm
2)
% S
urvi
val
Tum
or s
ize
(mm
2)
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
Control
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
LTX-315
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
aCTLA-4
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
aCTLA-4 + LTX-315
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
Controls (0/8)Cyclophosphamide (0/7)
LTX-315 (1/8)LTX-315 + cyclophosphamide (6/9)
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
days post tumor challenge
% S
urvi
val
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
Controls (0/9)Doxorubicin (0/9)
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
LTX-315 (0/8)LTX-315 + doxorubicin (5/10)
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
Iso Ctrl + PBS Iso Ctrl + LTX-315
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
aPD1 + PBS
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
8
Yamazaki et al., 2016, Cell Death and Differentiation
LTX-315 DEMONSTRATES SYNERGY with other standard of care cancer therapies
2
WITH CHEMOTHERAPY
AntiCTLA-4 Anti PD-1
D a y s a fte r tre a tm e n t
MC
A20
5 W
T
Tu
mo
r si
ze (
mm
2 )
0 1 0 2 0 3 00
1 0 0
2 0 0
3 0 0
4 0 0
Is o C tr l + P B S
Is o C tr l + L T X -3 1 5
α P D 1 + P B S
α P D 1 + L T X -3 1 5
LOW DOSE CYCLOPHOSHAMIDE (A2O B-LYMPHOMA) DOXORUBICIN (4T1 BREAST CANCER)
