Method of Selecting Patients for Treatment with a Combination of an AXL Inhibitor and an Immune Checkpoint Modulator
20230151100 · 2023-05-18
Assignee
Inventors
- James LORENS (Bergen, NO)
- David Micklem (Bergen, NO)
- Huiyu LI (Dallas, TX, US)
- Zhida LIU (Taiyuan City, CN)
- John MINNA (Dallas, TX, US)
- Rolf BREKKEN (Dallas, TX, US)
Cpc classification
C07K16/2863
CHEMISTRY; METALLURGY
C07K16/2866
CHEMISTRY; METALLURGY
A61K45/06
HUMAN NECESSITIES
A61K39/001102
HUMAN NECESSITIES
Y02A50/30
GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
G01N2800/52
PHYSICS
A61K31/502
HUMAN NECESSITIES
A61K2300/00
HUMAN NECESSITIES
A61K2300/00
HUMAN NECESSITIES
A61K31/502
HUMAN NECESSITIES
International classification
C07K16/28
CHEMISTRY; METALLURGY
A61K45/06
HUMAN NECESSITIES
Abstract
The invention relates to a method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: identifying subjects having an AXL-related disease characterised by the presence of cells having modified STK11 activity or expression; and, selecting thus identified subjects for treatment. The invention also relates to an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM) for use in the treatment of an AXL-related disease.
Claims
1. An AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM) for use in the treatment of an AXL-related disease, wherein the AXL-related disease is characterised by the presence of cells having modified STK11 activity or expression; and/or wherein the AXL-related disease is characterised by the presence of cells having increased KRAS activity or expression, and wherein the AXLi and ICM are for administration separately, sequentially or simultaneously.
2. An AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM) for use in the treatment of an AXL-related disease, said treatment comprising: identifying subjects that have previously been treated with an ICM and which did not respond to or benefit from treatment with the ICM; and, selecting thus identified subjects for treatment, wherein the AXLi and ICM are for administration separately, sequentially or simultaneously.
3. An AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM) for use in the treatment of an AXL-related disease, wherein the AXL-related disease is characterised by a reduced presence of CD8.sup.+ cells having TCF1 activity or expression, and wherein the AXLi and ICM are for administration separately, sequentially or simultaneously.
4. The AXLi and ICM for use according to any preceding claim, wherein the use further comprises the administration of a chemotherapeutic agent and/or radiotherapy.
5. The AXLi and ICM for use according to any preceding claim, wherein the AXL-related disease is further characterised by: i) the presence of cells having increased KRAS activity or expression; ii) the presence of cells having decreased p53 activity or expression; and/or iii) the presence of cells having increased AXL activity or expression.
6. The AXLi and ICM for use according to any preceding claim, wherein increased or decreased expression is assessed by: i) determining copy number of the gene encoding STK11, KRAS, or p53 relative to a control sample; and/or ii) determining the level of STK11, KRAS, or p53 protein or mRNA relative to a control sample.
7. The AXLi and ICM for use according to any preceding claim, wherein modified STK11 activity or expression is assessed by determining the presence or absence of a STK11 mutation and/or a STK11IP mutation.
8. The AXLi and ICM for use according to any preceding claim, wherein STK11 activity and/or expression is decreased relative to a control sample.
9. The AXLi and ICM for use according to claim 7 or claim 8, wherein the STK11 mutation or STK11IP mutation, is: i) a mutation in the nucleotide sequence encoding STK11 or STK11IP; ii) a mutation in a regulatory sequence controlling expression of the nucleotide sequence encoding STK11 or STK11IP; iii) a mutation in a nucleotide encoding a protein which interacts with the transcription product of the STK11 or STK11IP gene; iv) a mutation in the translation product of the STK11 or STK11IP gene; and/or v) a mutation in the transcription product of the STK11 or STK11IP gene.
10. The AXLi and ICM for use according to any one of claims 7-9, wherein STK11 mutation is an inactivating mutation, and/or the STK11IP mutation is an activating mutation.
11. The AXLi and ICM for use according to any preceding claim, wherein increased or decreased activity, expression, or population is determined in a sample derived from a subject.
12. The AXLi and ICM for use according to any preceding claim, wherein increased or decreased activity, expression, or population is determined relative to a control.
13. The AXLi and ICM for use according to any preceding claim, wherein the AXL-related disease is cancer, preferably a cancer selected from the group consisting of: lung cancer, non-small-cell lung cancer, breast cancer, melanoma, mesothelioma, acute myeloid leukemia (AML), myelodysplatic syndrome (MDS), pancreas cancer, kidney cancer, urothelial carcinoma, and glioblastoma.
14. The AXLi and ICM for use according to claim 13, wherein the cancer is lung cancer, preferably non-small-cell lung cancer.
15. The AXLi and ICM for use according to any preceding claim, further comprising administering to the subject a therapeutically effective amount of an AXL inhibitor (AXLi), an immune checkpoint modulator (ICM), and/or a chemotherapeutic agent and/or radiotherapy.
16. The AXLi and ICM for use according to any preceding claim, wherein the AXL inhibitor is selected from the group consisting of: 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7-(S)-pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7-(R)-pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(3-fluoro-4-(4-(pyrrolidin-1-yl)piperidin-1-yl)phenyl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N5-(7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-1-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N5-(7-(S)-pyrrolidin-1-yl-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(t-butoxycarbonylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(acetamido)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-((2R)-2-(methoxycarbonyl)pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4,4-difluoropiperidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-((methoxycarbonylmethyl)(methyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-((2R)-2-(carboxy)pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4-(ethoxycarbonyl)piperidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4-(carboxy)piperidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-((carboxymethyl)(methyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4-(ethoxycarbonylmethyl)piperazin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4-(carboxymethyl)piperazin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-1-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7s)-7-(di(cyclopropylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((2-methylpropyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((propyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(dipropylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(diethylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclohexylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclopentylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((1-cyclopentylethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(2-propylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((3,3-dimethylbut-2-yl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((cyclohexylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(cyclohexylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((5-chlorothien-2-yl)methyl)amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((2-carboxyphenyl)methyl)amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((3-bromophenyl)methyl)amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(dimethylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclobutylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(3-pentylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((2,2-dimethylpropyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(cyclopentylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((cyclopentylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(bicyclo[2.2.1]hept-2-en-5-ylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((bicyclo[2.2.1]hept-2-en-5-ylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(3-methylbutylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(3-methylbutyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(2-ethylbutylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(but-2-enylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(butyl(but-2-enyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N5-((7S)-7-(t-butoxycarbonylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-daimine; 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-pyrido[2′,3.:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(dimethylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(diethylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(dipropylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(cyclopropylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(3-methylbutyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclobutylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclohexylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((methylethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclopentylamino)-6,7,8,9-tetrahydro-5H-benzo[7]ann ulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; and 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(2-butylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; or pharmaceutically acceptable salts thereof.
17. The AXLi and ICM for use according to any preceding claim, wherein the AXL inhibitor is: i) Bemcentinib (BGB324/R428); or ii) selected from the group consisting of: dubermatinib (CAS No.1341200-45-0; UNII 14D65TV20J); gilteritinib (CAS No. 1254053-43-4; UNII 66D92MGC8M); cabozantinib (CAS No. 849217-68-1; UNII 1 C39JW444G); SG17079 (CAS No. 1239875-86-5); merestinib (CAS No. 1206799-15-6; UNII 50GS5K699E); amuvatinib (CAS No. 850879-09-3; UNII SO9S6QZB4R); bosutinib (CAS No. 380843-75-4; UNII 5018V4AEZ0); glesatinib (CAS No. 936694-12-1; UNII 7Q290XD98N); foretinib (CAS No. 849217-64-7; UNII 81FH7VK1C4); and, TP0903 (CAS No. 1341200-45-0); or iii) an anti-AXL antibody.
18. The AXLi and ICM for use according to any preceding claim, wherein the immune checkpoint modulator includes one or more immune checkpoint inhibitors (ICI), optionally wherein the immune checkpoint modulator is selected from the group consisting of: anti-CTLA-4 antibodies, anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-4-1BB antibodies, anti-OX-40 antibodies, anti-G ITR antibodies, anti-CD27 antibodies, anti-CD28 antibodies, anti-CD40 antibodies, anti-LAGS antibodies, anti-ICOS antibodies, anti-TWEAKR antibodies, anti-HVEM antibodies, anti-TIM-1 antibodies, anti-TIM-3 antibodies, anti-VISTA antibodies, and anti-TIGIT antibodies.
19. The AXLi and ICM for use according to any preceding claim, wherein the immune checkpoint modulator is selected from the group consisting of: anti-CTLA-4 antibodies, anti-PD-1 antibodies, and anti-PD-L1 antibodies.
20. The AXLi and ICM for use according to any preceding claim, wherein the immune checkpoint modulator includes, or is: pembrolizumab; ipilimumab; ipilimumab and nivolumab; ipilimumab and pembrolizumab; tremelilumab and durvalumab.
21. The AXLi and ICM for use according to any one of claims 4 to 20, wherein the chemotherapeutic agent is a chemotherapeutic agent which induces immunogenic cell death of cancer cells.
22. The AXLi and ICM for use according to any one of claims 4 to 21, wherein the chemotherapeutic agent is a chemotherapeutic agent which induces an immune response in the subject, optionally wherein the chemotherapeutic agent is a chemotherapeutic agent which induces a type I interferon response in the subject.
23. The AXLi and ICM for use according to any one of claims 4 to 22, wherein the chemotherapeutic agent is an anthracycline, optionally wherein the anthracycline is doxorubicin, daunorubicin, epirubicin, idarubicin, mitoxantrone, or valrubicin, preferably doxorubicin.
24. A method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: identifying subjects having an AXL-related disease characterised by the presence of cells having modified STK11 activity or expression; and, selecting thus identified subjects for treatment.
25. A method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: identifying subjects having an AXL-related disease characterised by the presence of cells having increased KRAS activity or expression; and, selecting thus identified subjects for treatment.
26. A method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: identifying subjects that have previously been treated with an immune checkpoint modulator (ICM) and which did not respond to or benefit from treatment with the ICM; and, selecting thus identified subjects for treatment.
27. A method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: identifying subjects having an AXL-related disease characterised by a reduced presence of CD8.sup.+ cells having TCF1 activity or expression; and, selecting thus identified subjects for treatment.
28. A method of increasing a population of desired T cells in a subject comprising treating the subject with an AXL inhibitor (AXLi).
29. The method of claim 28, wherein the desired T cell population is a CD8+ T cell population.
30. A method of treating an AXL-related disease in a subject with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: (i) administering a combination of the AXLi and the ICM to the subject; (ii) administering the AXLi to the subject, wherein the ICM has been, is, or will be, administered to the subject; or (iii) administering the ICM to the subject, wherein the AXLi has been, is, or will be, administered to the subject; wherein the AXL-related disease is characterised by: the presence of cells having decreased STK11 activity or expression; and/or the presence of cells having a STK11 mutation and/or a STK11IP mutation; optionally wherein the subject has been selected for treatment using a method as defined in any one of claims 1-23.
31. A method of treating an AXL-related disease in a subject with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: (i) administering a combination of the AXLi and the ICM to the subject; (ii) administering the AXLi to the subject, wherein the ICM has been, is, or will be, administered to the subject; or (iii) administering the ICM to the subject, wherein the AXLi has been, is, or will be, administered to the subject; wherein the AXL-related disease is characterised by: the presence of cells having increased KRAS activity or expression; and/or the presence of cells having a KRAS mutation; optionally wherein the subject has been selected for treatment using a method as defined in any one of claims 1-23.
32. The method of claim 30 or 31, wherein the subject is treated with a combination of an AXL inhibitor (AXLi), an immune checkpoint modulator (ICM), and a chemotherapeutic agent and/or radiotherapy.
33. A method of prognosing susceptibility of a subject to treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: determining: i) the presence or absence of a STK11 mutation and/or STK11I P mutation; and/or ii) the level of STK11 activity or expression in the subject or a sample derived from the subject; wherein the presence of a STK11 mutation and/or presence of a STK11I P mutation, and/or a modified level of STK11 activity or expression is indicative of susceptibility to treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), preferably wherein the modified level of STK11 activity or expression is a decreased level of STK11 activity or expression.
34. A method of prognosing susceptibility of a subject to treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: determining: i) the presence or absence of a KRAS mutation; and/or ii) the level of STK11 activity or expression in the subject or a sample derived from the subject; wherein the presence of a KRAS mutation, and/or an increased level of STK11 activity or expression is indicative of susceptibility to treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM).
35. A method of treating an AXL-related disease in a subject in need of such treatment, the method comprising administering to the subject a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), wherein the AXLi and ICM may be administered to the subject simultaneously, separately or sequentially.
36. An AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM) for use in the treatment of an AXL-related disease in a subject, wherein the AXLi and ICM may be administered to the subject simultaneously, separately or sequentially.
Description
FIGURES
[0982]
[0983]
[0984]
[0985]
[0986]
[0987]
[0988]
[0989]
[0990]
[0991] (B) Shows tumor growth of C57BL/6J mice (n=5) inoculated with 1×10.sup.6 KPL9-3-1 tumor cells with interferon alpha receptor blocking antibody treated with either BGB324 (50 mg/kg, twice daily), and PD-1 (10 mg/kg, day 7, 10, 14), or corresponding IgG and vehicle started at day 7 after tumor inoculation. Tumor growth was measured every 3 days.(C-D) shows the abundance of TCF1.sup.+PD-1.sup.+ cells among gated CD8.sup.+ TILs (per mm.sup.3 of tumor) at day 7 after treatment started. (E) Shows the mean fluorescent intensities (MFIs) of TCF1.sup.+ cells among gated CD8.sup.+ OT-1 cells. Bone marrow dendritic cells (BMDC) were co-cultured with isolated CD8.sup.+ T cells stimulated with ovalbumin.(F) Shows MFIs of TCF1.sup.+ cells among gated CD8.sup.+ OT-1 cells. (H) Shows the abundance of TCF1.sup.+PD-1.sup.+ cells among gated CD8.sup.+ TILs (per mm.sup.3 of tumor) at 48 h after with or without IFNα (200 ng) intratumoral injection.
TABLE-US-00001 SEQUENCES [10C9 Heavy CDR1] SEQ ID NO. 1 DYNFTRYYIH [10C9 Heavy CDR2] SEQ ID NO. 2 WIYPGTGDSKYNEKFKG [10C9 Heavy CDR3] SEQ ID NO. 3 NGNYWYFDV [10C9 Light CDR1] SEQ ID NO. 4 RSSKSLLHSNGNTYLY [10C9 Light CDR2] SEQ ID NO. 5 RMSNLAS [10C9 Light CDR3] SEQ ID NO. 6 MQHREYPFT [10G5 Heavy CDR1] SEQ ID NO. 7 GYSFTDFYIN [10G5 Heavy CDR2] SEQ ID NO. 8 RIFPGGDNTYYNEKFKG [10G5 Heavy CDR3] SEQ ID NO. 9 RGLYYAMDY [10G5 Light CDR1] SEQ ID NO. 10 RSSQSLVHSNGIPYLH [10G5 Light CDR2] SEQ ID NO. 11 RVSNRFS [10G5 Light CDR3] SEQ ID NO. 12 SQGTHVPPT [hu10G5 VH(GH1)] SEQ ID NO. 13 EVQLVQSGAGLVQPGGSVRLSCAASGYSFTDFYIN WVRQAPGKGLEWIARIFPGGDNTYYNEKFKGRFTL SADTSSSTAYLQLNSLRAEDTAVYYCARRGLYYAM DYWGQGTLVTVSS [hu10G5 VH(GH2)] SEQ ID NO. 14 EVQLVESGGGLVQPGGSLRLSCAASGYSFTDFYIN WVRQAPGKGLEWVARIFPGGDNTYYNEKFKGRFTL SADTSKSTAYLQMNSLRAEDTAVYYCARRGLYYAM DYWGQGTLVTVSS [hu10G5 VL(GL1)] SEQ ID NO. 15 DIQMTQSPSSLSASVGDRVTITCRSSQSLVHSNGI PYLHWYQQKPGKAPKLLIYRVSNRFSGVPSRFSGS GSGTDFTLTISSLQPEDFATYYCSQGTHVPPTFGQ GTKVEIK [hu10G5 VL(GL2)] SEQ ID NO. 16 DIQMTQSPSSLSASVGDRVTITCRSSQSLVHSNGI PYLHWYQQKPGKAPKLLIYRVSNRFSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCSQGTHVPPTFGQ GTKVEIK [10G5 GH1 Heavy chain] SEQ ID NO. 17 EVQLVQSGAGLVQPGGSVRLSCAASGYSFTDFYIN WVRQAPGKGLEWIARIFPGGDNTYYNEKFKGRFTL SADTSSSTAYLQLNSLRAEDTAVYYCARRGLYYAM DYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT KVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPGK [10G5 GH2 Heavy chain] SEQ ID NO. 18 EVQLVESGGGLVQPGGSLRLSCAASGYSFTDFYIN WVRQAPGKGLEWVARIFPGGDNTYYNEKFKGRFTL SADTSKSTAYLQMNSLRAEDTAVYYCARRGLYYAM DYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT KVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPGK [10G5 GL1 Light chain] SEQ ID NO. 19 DIQMTQSPSSLSASVGDRVTITCRSSQSLVHSNGI PYLHWYQQKPGKAPKLLIYRVSNRFSGVPSRFSGS GSGTDFTLTISSLQPEDFATYYCSQGTHVPPTFGQ GTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCL LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD STYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC [10G5 GL2 Light chain] SEQ ID NO. 20 DIQMTQSPSSLSASVGDRVTITCRSSQSLVHSNGI PYLHWYQQKPGKAPKLLIYRVSNRFSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCSQGTHVPPTFGQ GTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCL LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD STYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC
EXAMPLES
[0992] The following biological examples are provided by way of illustration, not limitation. In the following biological examples, 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazi n-3-yl)-N.sup.3-((7-(S)-pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine, which is a compound of formula (I), as defined above, is designated in the following examples and the Figures as “BGB324” or Bemcentinib. In the following Examples the immune checkpoint modulator is an anti-PD-1 antibody, as indicated by name in the relevant examples.
[0993] STK11/p53/KRAS Mutation Profile is Indicative of Response to AXLi+ICM Combination Therapy in Human NSCLC Patients
[0994] Whole Exome Sequence analysis of biopsies collected from responder subjects in a phase II non-small cell lung cancer trial of AXLi (Bemcentinib; BGB324) in combination with ICM (pembrolizumab) identified five subjects with mutations in STK11 and/or STK11IP (STK11 interacting protein). In each case, these responder subjects also had mutations in KRAS and/or p53. Response to the combination therapy was independent of PDL1 status. These findings were unexpected in view of literature reports that subjects with STK11 mutations respond poorly to PD-1 inhibition.
[0995] Formalin-fixed paraffin-embedded (FFPE) tumor tissue from 20 patients was subjected to whole exome sequencing, with variants identified using the Pisces 5.2.5.20 (Illumina) variant calling suite (Dunn et al, 2019). The following subjects had identified variants in either STK11 or STK11IP:
TABLE-US-00002 Patient number mutations response PD-L1 status 234005 STK11IP, KRAS, P53 Partial Negative Response (PR) 234008 STK11, STK11IP, PR Negative P53, PTEN 237001 STK11IP, KRAS PR High 211105 STK11, P53 Stable Weak positive Disease (SD) 211002 STK11IP, BRAF, P53 PR Weak Positive
[0996] A sixth subject (subject number 211003) with a variant STK11 sequence was also identified, however this subject was non-evaluable for response. Predicted severity of the identified mutations was assessed using mutationassessor.org (polyphen-2)—a computational system for predicting the functional impact of protein missense mutations based on phenomenological analysis of information extracted from protein family alignments of large numbers of homologous sequences grouped into aligned sets (families and subfamilies) and the 3D structures of sequence homologs (Reva et al, 2011). The identified mutations and predicted functional impact were as follows:
TABLE-US-00003 Predicted Patient severity/functional number mutations impact 211003 STK11 - L160P High Probably-damaging 234005 STK111P - LG334FW Low + Medium Probably-damaging 234008 STK11 - LD140PY Medium + Neutral STK11IP - W162C Probably-damaging Low Probably-damaging 237001 STK11IP - R1065Q Medium Probably-damaging 211105 STKIP - D115V Medium Probably-damaging 211002 STK11IP - E19V Low Probably-damaging
[0997] Patient Results
[0998] Patient 234008, a 79-year-old male, initially achieved a partial response to carboplatin and paclitaxel first-line therapy for 22 months before developing progression of lung and adrenal metastases as well as bone marrow metastasis and enrolling in the study. The patient's tumor biopsy was negative for PD-L1 expression but showed AXL expression in both tumor and immune cells. The patient went on to achieve a partial response to the bemcentinib/pembrolizumab combination lasting 11 months from the start of treatment with a maximum target lesion shrinkage of 50.6%. The patient was still alive 2 years after starting treatment.
[0999] Patient 211105, a 73-year-old male, was treated with first-line pemetrexed and cisplatin and progressed after 8 months to pembrolizumab monotherapy, where he experienced clinical benefit for 18 months before developing progression of lymph node and chest wall metastases. At the time of screening, the patient's tumor biopsy was PD-L1 weak positive and showed strong expression of AXL in tumor-infiltrating immune cells. The patient experienced clinical benefit from the study drug combination, achieving stable disease for 5.4 months.
[1000] Treatment in Patients who Have Previously Received an Anti-PD(L)1 Monoclonal Antibody as Monotherapy or Combination Therapy
[1001] Patients were recruited for a phase II trial for the combination treatment of an AXL inhibitor and an anti-PD1/L1 mAb therapy. Patients had previously been treated with a mono therapy PD-L1 or PD-1 inhibitor and had previously demonstrated disease control on this treatment.
[1002] The previous therapies include standard dosing regimes such as: [1003] Pembrolizumab (200 mg every 3 weeks, administered over a 30-minute infusion, until disease progression or unacceptable toxicity, or up to 24 months without disease progression; [1004] Pembrolizumab in combination with pemetrexed and platinum chemotherapy (200 mg IV q3Weeks OR 400 mg q6Weeks until disease progression, unacceptable toxicity, or up to 24 months without disease progression); [1005] Atezolizumab (840 mg IV q2Weeks or 1200 mg IV q3Weeks or 1680 mg IV q4Weeks until disease progression or unacceptable toxicity); [1006] Atezolizumab in combination with 1) bevacizumab, paclitaxel, and carboplatin or 2) paclitaxel protein-bound and carboplatin uses the same dosages; [1007] Nivolumab (240 mg IV q2Weeks or 480 mg IV q4Weeks continue until disease progression or unacceptable toxicity) [1008] Nivolumab in combination with Ipilimumab (Nivolumab 3 mg/kg IV q2Weeks plus Ipilimumab 1 mg/kg IV q6Weeks, continue until disease progression, unacceptable toxicity, or up to 24 months without disease progression; or [1009] Nivolumab in combination with Ipilimumab and platinum chemotherapy (Nivolumab 360 mg/kg IV q3Weeks PLUS Ipilimumab 1 mg/kg IV q6Weeks PLUS Histology-based platinum doublet chemotherapy q3Weeks for 2 cycles continue until disease progression, unacceptable toxicity, or up to 24 months without disease progression.
[1010] The patients all had progressive disease upon screening. 21 patients were screened and 16 patients were enrolled on the trial. Patient demographics are shown below:
TABLE-US-00004 Patient disposition Number of Patients Screened 21 Enrolled 16 Evaluable* 15 Ongoing 3 *with at least 1 post-baseline scan assessment.
TABLE-US-00005 Patient demographics N (%) Age Median 64.5 Range 40-76 Eastern Cooperative 0 6 (38) Oncology Group 1 10 (63) (ECOG) at screen Sex Female 3 (19) Male 13 (81) Smoking status Smoker 6 (38) Ex-smoker 8 (50) Never smoked 0 (0) Unknown 1 (6)
TABLE-US-00006 Disease Mutations N (%) None 13 (81) KRAS 2 (13) BRAF 1 (6)
[1011] 25% of the patients enrolled on the trial were PD-L1 negative (tumor proportion score (TPS)<1%), 42% PD-L1 positive (TPS 1-49%) and 33% PD-L1 strong positive (TPS>50%).
[1012] The results are shown in
[1013]
[1014] Results
[1015] This demonstrates that a combination of an AXL inhibitor and an Anti-PD1 antibody therapy has surprising results in patients that have previously been treated with a mono immunotherapy but have since suffered disease progression.
[1016] STK11 Mutation Abrogates Anti-PD-1 Efficacy
[1017] Cell Culture
[1018] Kras.sup.G12D, p53.sup.−/− mutant (KP9-1) or Kras.sup.G12D, p53.sup.−/− STK11.sup.−/− mutant (KPL9-3) mouse lung cancer-derived cell lines were prepared using standard techniques known in the art. KP9-1 or KPL9-3 cells were propagated at sub-confluence and split on a regular basis.
[1019] Subcutaneous Tumor Inoculation
[1020] Each animal was weighed before cell implantation. Injection of cells was performed after anesthetizing of the mice. The cell lines discussed above were injected subcutaneously into immune-competent mice (C57Bl/6 strain).
[1021] Mice were treated with either placebo or anti-PD1 therapeutic (200 μg/mouse, intraperitoneal, twice a week for two weeks).Tumor volume was assessed using hand held callipers and mouse weight were followed.
[1022] Tumors derived from the Kras.sup.G12D, p53.sup.−/− STK11.sup.−/− mutant cells were resistant to anti-PD1 therapy. Whereas, the tumors derived from the P53 −/− mic were sensitive to anti-PD-1 therapy. The results are shown in
[1023] Results
[1024] These results show that the STK11 mutation reduces the efficacy of anti-PD-1 treatment.
[1025] AXL Inhibition Sensitizes STK11 Mutant Cells to Immunotherapy
[1026] The Kras.sup.G12D, p53.sup.−/− STK11.sup.−/− mutant (KPL9-3) mouse lung cancer-derived cells described above were implanted subcutaneously in immune-competent mice (C57Bl/6 strain) using the same protocol as described above.
[1027] Mice were treated with either placebo, anti-PD1 therapeutic (10 mg/kg, intraperitoneal, twice a week for two weeks), BGB324 (50 mg/kg, oral gavage, twice daily) or a combination of both drugs. Tumor volumes were measured.
[1028] Tumors were resistant to anti-PD1 therapy and BGB324 as monotherapies, but responded to the combination treatment. Tumor volumes are shown in
[1029] Results
[1030] These results surprisingly show that an AXL inhibitor is able to sensitise the STK11 mutant cells to treatment with an immune checkpoint inhibitor.
[1031] Combination Treatment of an AXL Inhibitor and an Immune Checkpoint Inhibitor in Human Non-Small Cell Lurid Cancer.
[1032] Cell Culture
[1033] Human non-small cell lung cancer (NSCLC) cells Kras.sup.G12D, p53 WT STK11.sup.−/− mutant human A549 lung cancer cells were cultured using standard protocols.
[1034] Subcutaneous Implantation
[1035] Each animal was weighed before cell implantation. Injection of the cells was performed after anesthetizing of the mice. Kras.sup.G12D, p53 WT STK11.sup.−/− mutant human A549 lung cancer cells were implanted subcutaneously in humanized immune-deficient mice.
[1036] Mice were treated with either placebo, anti-PD1 therapy (10 mg/kg, intraperitoneal, 3 dosages), BGB324 (50 mg/kg, oral gavage, twice daily for 3 weeks) or a combination of both drugs.
[1037] Tumor volumes were measured and the results are shown in
[1038] Results
[1039] These results confirm that an AXL inhibitor is able to sensitise the STK11 mutant cells to treatment with an immune checkpoint inhibitor.
[1040] The results in the Examples above demonstrate that treating a subject with a combination of an AXL inhibitor and an immune checkpoint modulator, wherein the subject has modified STK11 activity or expression, will result in a surprisingly good response in this difficult-to-treat patient group.
[1041] STK11/LKB1 Mutated NSCLC Lacks Anti-PD-1 Treatment Responsive T Cells in the TME.
[1042] Method
[1043] For subcutaneous allografts, 1×10.sup.6 cells in 100 ul phosphate buffered saline (PBS) were injected into the right dorsal flanks of 6-8 wks old mice. For xenografts growing on humanized mice, 1.5×10.sup.6 cells in 100 ul phosphate buffered saline (PBS) were injected into the right dorsal flanks of reconstituted humanized mice.
[1044] Tumor bearing mice were randomly assigned into treatment groups when the tumor grew to around 100-150 mm.sup.3. For each treatment group, 5 mice were assigned. 10 mg/kg anti-PD-1 (BioXCell, Cat #BE0146) treatment was given intraperitoneally on day 0, day 4 and day 7 after innoculation. The control group was treated with 10 mg/kg rat IgG2a isotype control (BioX Cell, Cat #BE0089) at the same day of PD-1 treatment, intraperitoneally. For the tumor growth measurement experiment, mice were treated for 3 weeks. For tumor microenvironment (TME) analysis, mice were treated for 7 days, when the tumor sizes in each treatment group are within 2-fold differences. Tumor volumes were measured by length (a), width (b) and height (h) in every 3 days and calculated as tumor volume=abh.
[1045] KP9-3 and KPL9-3-1 allografts were harvested 14 days after subcutaneous implantation (average tumor size ˜200 mm.sup.3). 10 tumors harvested from each individual mouse within the same group were pooled together as one sample. The CD8.sup.+ T cell status and composition differences caused by the LKB1 mutation were analysed by aggregating CD8.sup.+ T cells from KP9-3 and KPL9-3-1 tumors and then clustering into 4 clusters annotated with appropriate markers.
[1046] Results
[1047] Subcutaneous KP9-3 tumors grew faster than KPL9-3-1 tumors in NSG mice, while KPL9-3-1 grew faster than KP9-3 when injected into C57BL/6J mice. The results in
[1048] Based on the percentage of CD8.sup.+ T cells from each cluster, loss of STK11/LKB1 resulted in a more suppressive CD8.sup.+ T cell composition. In
[1049] The results were confirmed by comparing TCF1.sup.+PD-1.sup.+ CD8.sup.+ T cells in KP9-3 and KPL9-3-1 tumors through flow cytometry and IHC. The results are shown in
[1050] Bemcentinib Mediated Axl Inhibition Sensitizes LKB1 Mutant Tumors to Anti-PD-1 Therapy.
[1051] Methods
[1052] KPL9-3-1 allograft were implanted into mice as described above. 5 mice were assigned into each treatment group. 10 mg/kg anti-PD-1 (BioXCell, Cat #BE0146) treatment was given intraperitoneally on day 0, day 4 and day 7 after treatment. Control group of PD-1 treatment was treated with 10 mg/kg rat IgG2a isotype control (BioX Cell, Cat #BE0089) at the same day of PD-1 treatment, intraperitoneally. BGB324 were given through oral gavage twice daily, with a dose of 50 mg/kg.
[1053] For tumor growth measurement experiment, mice were treated for 3 weeks. For tumor microenvironment analysis, mice were treated for 7 days, when the tumor sizes in each treatment group are within 2-fold differences.Seven days after tumor inoculation, mice were treated with either BGB324 or anti-PD-1 alone, or in combination.
[1054] The immune landscape of the tumors from each treatment group was assessed using scRNAseq of immune cells. Sequenced scRNA-seq samples were processed through Cellranger pipelines (v3.1.0). Cellranger count was used to align reads to mouse reference genome (mm10, 2020-A, from 10× Genomics) and generate single cell feature counts for single library.
[1055] For scTCR-seq data, TCR reads were aligned to reference genome and TCR annotation was performed using the 10×cellranger vdj pipeline with provided reference (cellranger-vdj-GRCm38-alts-ensemb1-4.0.0). Overall, 94% of T cells in scRNA-seq data were assigned a TCR and more than 70% had at least one full-length productive CDR3 for both TRA and TRB. The clonotype of each T cell was represented by the beta CDR3 sequences, and clone sizes ranged from 1 cell to 580 cells. Cellranger aggr were applied next to aggregate each sample library for grouped analysis with same effective sequencing depth. Seurat (3.2.1) package was used for downstream analysis.
[1056] To determine the potential lineage differentiation between those T cell populations with high TCR sharing. Monocle (version 2.0) (PMID: 24658644) was used to investigate transcriptional and developmental trajectories concerning different CD8+ or CD4+ T cell clusters. The data of raw counts together with cluster annotations were taken as input to monocle, and the default parameters were set to run data normalization and dimension reduction. Next, Monocle leaned the kinetics of gene expression by using the reversed graph embedding approach and places each cell along an inferred pseudotime trajectory. According to the assumption that the trajectory has a tree structure, functional “State” is identified based on the segment of the tree-like structure.
[1057] Results
[1058] Neither single treatment resulted in tumor growth control. However, the combination of BGB324 with anti-PD-1 treatment showed a synergistic effect with sustained control of tumor progression (
[1059] BGB324 treatment alone or in combination with anti-PD-1 increased the infiltration of TCF1+PD-1.sup.+ CD8.sup.+ T cells (
[1060] Immunohistochemistry (IHC) analysis demonstrated that while Axl inhibition alone enhanced TCF1.sup.+PD-1.sup.+ CD8.sup.+ T cell presence in the tumor, combination with anti-PD-1 was required to break the exclusion of TCF1.sup.+CD8.sup.+ T cells from tumor islands (
[1061] To further dissect the dynamics changes in the KPL9-3-1 TME after treatment, scRNAseq with paired TCR sequencing were performed for four pooled samples from each treatment group annotated with representative markers. Sub-clustering of CD8.sup.+ T cells revealed eight defined annotated sub-populations.
[1062] Treatment enriched cells in each cluster were calculated and compared based on the number of cells observed divided by expected number of cells in each cluster (
[1063] BGB324 treated tumors enriched CD8.sup.+ T cells expressing unique TCRs (clonal expanded) significantly, with the stem like T cells and exhaustive effector T cells also enriched. Combination therapy showed a trend of enrichment of clonal expanded and exhaustive effector CD8.sup.+ T cells.
[1064] This analysis demonstrates that stem-like T cells correlate the most with clonal expanded T cells, which develop into proliferating and exhaustive effector T cells capable of performing direct tumor cell kill (
[1065] Bemcentinib-Induced Type I Interferon Secretion Expands TCF1.sup.+PD-1.sup.+CD8.sup.+ T Cells
[1066] Methods
[1067] Bone marrow derived BMDCs (2×10.sup.5) were co-cultured with 40 g irradiated KPL9-3-1 tumor cells (2×10.sup.6), with or without BGB324 treatment (40 nM). After 24 h, the cell supernatant was collected for analysis. The concentration of IFN8 was measured by VeriKine-HS Mouse IFN Beta Serum ELISA kit (PBL Assay Science, Cat #42410), or VerKine Human IFN Beta ELISA kit (PBL Assay Science, Cat #414101) in accordance with the manufacturer's instructions. The plates were visualized by adding 100 μL of TMB solution and read at 450 nm using the SPECTROstarNano (BMG LABTECH).
[1068] The cells were treated with BGB324 or DMSO.
[1069] To determine the contribution of type I interferon secretion to BGB324 sensitization of KPL9-3-1 tumors to anti-PD-1 treatment the IFNα receptor was inhibitred pharmacologically.
[1070] Results
[1071] BMDCs treated with BGB324 showed increased secretion of Interferon type 1β (IFNβ) (
[1072] Blocking IFNα receptor intratumorally abrogated the efficacy of combined Axl and PD-1 inhibition (
[1073] IFNα receptor blockade diminished the infiltration of TCF1.sup.+PD-1.sup.+CD8.sup.+ T cells in the TME (
[1074] Bemcentinib-Induced Type I Interferon Secretion Expands TCF1.sup.+PD-1.sup.+CD8.sup.+ T Cells in Axl-Deficient BMDCs
[1075] Methods
[1076] To demonstrate the importance of Type I IFN response, the level of TCF1 expression in OT-1 CD8 T cells stimulated with OVA and cultured with BMDCs +/− IFNα was determined by flow cytometry. Tumor tissues were excised and digested with 2 mg/mL Collagenase A (Sigma, Cat #SCR136) and 1 mg/ml DNase I (Roche, Cat #11284932001) under 37° C., 150 rpm shaking speed for 45 min. Digested materials then were transferred to a 70 μm cell strainer to remove clumped cells. Digested cells were washed twice with FACs buffer and ready for flow cytometry analysis.
[1077] Results
[1078] The results are shown in
[1079] Conclusion
[1080] Taken together, these results demonstrate that increased IFN I secretion as a result of Axl inhibition is critical for inducing TCF1.sup.+ PD-1.sup.+CD8.sup.+ T cell expansion in KPL9-3-1 tumors to overcome anti-PD-1/PD-L1 resistance.
[1081] Mutation of KRAS and STK11 Results in High AXL in Tumor Immune Cells
[1082] Methods
[1083] Tissue microarray analysis of 62 NSCLC patients was performed. 7 patients had mutation of both KRAS and STK11. 55 were KL wild type.
[1084] Tissue microarrays (TMAs) were generated with surgical resected non-small cell lung carcinoma tumor samples (TMA3). IHC staining was performed on 4-μm thick TMA sections in a Leica Bond RX automated strainer (Leica Biosystems). The antigen retrieval was performed with Bond ER Solution #1 (Leica Biosystems) equivalent to citrate buffer, pH 9.0 for 20 min at 100° C. The sections were then incubated with anti-AXL antibody (rabbit monoclonal, Cell Signaling clone C89E7); 1:300 dilution (1 μg/ml). The antibody was detected using a Bond Polymer Refine Detection kit (Leica Biosystems) with diaminobenzidine (DAB) as the chromogen. All the slides were counterstained with hematoxylin, dehydrated, and cover slipped. Tonsil and normal colon sections were used as external positive controls. Each case was analyzed using standard microscopy by two pathologists in immune cells and reported as percentage of the tumor area occupied with immune cell with cytoplasmic and/or membrane expression.
[1085] Results
[1086] All the KL mutant patients had high AXL in tumor immune cells; whereas, in KL wild type patients, only 20 of the 55 patients displayed high AXL.
[1087] Conclusion
[1088] These results demonstrate that KRAS and STK11 mutations are good predictors of AXL activity. Patients with KRAS and/or STK11 mutations will benefit from treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM).
[1089] Statements of Disclosure
[1090] The following numbered statements, outlining aspects of the present disclosure, are part of the description.
[1091] Methods of Selecting a Subject for Treatment
[1092] 101. A method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: identifying subjects having an AXL-related disease characterised by the presence of cells having modified STK11 activity or expression; and, selecting thus identified subjects for treatment.
[1093] 102. A method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: identifying subjects having an AXL-related disease characterised by the presence of cells having increased KRAS activity or expression; and, selecting thus identified subjects for treatment.
[1094] 103. A method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: identifying subjects that have previously been treated with an immune checkpoint modulator (ICM) and which did not respond to treatment with the ICM; and, selecting thus identified subjects for treatment.
[1095] 104. The method of statement 103, wherein the method further comprises: identifying subjects having an AXL-related disease characterised by the presence of cells having modified STK11 activity or expression; and, selecting thus identified subjects for treatment.
[1096] 105a. A method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi), an immune checkpoint modulator (ICM), and a chemotherapeutic agent and/or radiotherapy, the method comprising: identifying subjects having an AXL-related disease characterised by the presence of cells having modified STK11 activity or expression; and, selecting thus identified subjects for treatment.
[1097] 105b. A method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi), an immune checkpoint modulator (ICM), and a chemotherapeutic agent and/or radiotherapy, the method comprising: identifying subjects having an AXL-related disease characterised by the presence of cells having increased KRAS activity or expression; and, selecting thus identified subjects for treatment.
[1098] 106. The method of statement 105a or statement 105b, wherein the method further comprises: identifying subjects that have previously been treated with a combination of an immune checkpoint modulator (ICM) and chemotherapeutic agent and/or radiotherapy, wherein treatment with the combination of immune checkpoint modulator (ICM) and chemotherapeutic agent and/or radiotherapy did not provide any additional benefit as compared to treatment with the chemotherapeutic agent and/or radiotherapy alone; and, selecting thus identified subjects for treatment.
[1099] 107. A method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi), an immune checkpoint modulator (ICM), and a chemotherapeutic agent and/or radiotherapy, the method comprising: identifying subjects that have previously been treated with a combination of an immune checkpoint modulator (ICM) and chemotherapeutic agent and/or radiotherapy, wherein treatment with the combination of immune checkpoint modulator (ICM) and chemotherapeutic agent and/or radiotherapy did not provide any additional benefit as compared to treatment with the chemotherapeutic agent and/or radiotherapy alone; and, selecting thus identified subjects for treatment.
[1100] 108. The method of statement 107, wherein the method further comprises: identifying subjects having an AXL-related disease characterised by the presence of cells having decreased STK11 activity or expression; and, selecting thus identified subjects for treatment.
[1101] Patient Sub-Group Definition
[1102] 109. The method of any preceding statement, wherein the method comprises: identifying subjects wherein the AXL-related disease is further characterised by the presence of cells having increased KRAS activity or expression; and, selecting thus identified subjects for treatment.
[1103] 110. The method of any preceding statement, wherein the method comprises: identifying subjects wherein the AXL-related disease is further characterised by the presence of cells having decreased p53 activity or expression; and, selecting thus identified subjects for treatment.
[1104] 111. The method of any preceding statement, wherein the method comprises: identifying subjects wherein the AXL-related disease is further characterised by increased activity or expression of AXL as compared to a control.
[1105] Assessing Increased/Decreased Expression
[1106] 112. The method of any preceding statement, wherein increased or decreased expression is assessed by determining copy number of the gene encoding STK11, KRAS, or p53 relative to a control sample, wherein an increase in the copy number indicates an increased level of expression and a decrease in the copy number indicates a decreased level of expression.
[1107] 113. The method of any preceding statement, wherein increased or decreased expression is assessed by determining the level of STK11, KRAS, or p53 protein or mRNA relative to a control sample.
[1108] 114. The method of any preceding statement, wherein modified STK11 activity or expression is assessed by determining the presence or absence of a STK11 mutation and/or a STK11IP mutation.
[1109] 115. The method of any one of statements 110-114, wherein increased KRAS activity or expression is assessed by determining the presence of absence of a KRAS mutation.
[1110] 116. The method of any one of statements 111-115, wherein decreased p53 activity or expression is assessed by determining the presence of absence of a p53 mutation.
[1111] 117. The method of any one of statements 114-116, wherein the STK11 mutation, STK11IP mutation, KRAS mutation, and/or p53 mutation is a mutation selected from: [1112] (i) a mutation in the nucleotide sequence encoding STK11, STK11IP, KRAS, or p53; [1113] (ii) a mutation in a regulatory sequence controlling expression of the nucleotide sequence encoding STK11, STK11IP, KRAS, or p53; [1114] (iii) a mutation in a nucleotide encoding a protein which interacts with the transcription product of the STK11, STK11IP, KRAS, or p53 gene.
[1115] 118. The method of any one of statements 114-117, wherein the STK11 mutation, STK11IP mutation, KRAS mutation, and/or p53 mutation is a mutation in the translation product of the STK11, STK11IP, KRAS, or p53 gene.
[1116] 119. The method of any one of statements 114-118, wherein the STK11 mutation, STK11IP mutation, KRAS mutation, and/or p53 mutation is a mutation in the transcription product of the STK11, STK11IP, KRAS, or p53 gene.
[1117] 120. The method of any one of statements 114-119, wherein the STK11 mutation is an inactivating mutation.
[1118] 121. The method of any one of statements 114-120, wherein the STK11IP mutation is an activating mutation.
[1119] 122. The method of any one of statements 115-121, wherein the KRAS mutation is an activating mutation.
[1120] 123. The method of statement 122, wherein the KRAS mutation is a mutation at position G12, optionally wherein the KRAS mutation is a G12D mutation.
[1121] 124. The method of any one of statements 116-123, wherein the p53 mutation is an inactivating mutation.
[1122] 125. The method of any preceding statement, wherein increased or decreased activity or expression is determined in a sample derived from a subject.
[1123] 126. The method of any preceding statement, wherein increased or decreased activity or expression is determined relative to a control.
[1124] 127. The method of statement 126, wherein the control is healthy tissue, preferably of the same tissue type as the AXL-related disease.
[1125] AXL-Related Disease
[1126] 128. The method of any preceding statement, wherein the AXL-related disease is a proliferative disease.
[1127] 129. The method of any preceding statement, wherein the AXL-related disease is a neoplastic disease.
[1128] 130. The method of any preceding statement, wherein the AXL-related disease is a solid tumour.
[1129] 131. The method of any preceding statement, wherein the AXL-related disease is cancer.
[1130] 132. The method of statement 131, wherein the cancer is selected from the group consisting of: lung cancer, non-small-cell lung cancer, breast cancer, melanoma, mesothelioma, acute myeloid leukemia (AML), myelodysplatic syndrome (MDS), pancreas cancer, kidney cancer, urothelial carcinoma, and glioblastoma.
[1131] 133. The method of statement 131, wherein the cancer is lung cancer.
[1132] 134. The method of statement 131, wherein the cancer is non-small-cell lung cancer (NSCLC).
[1133] 135. The method of any preceding statement, wherein the AXL-related disease does not respond to or benefit from treatment with an immune checkpoint modulator (ICM) when administered alone or as part of a treatment regime that does not include an AXLi.
[1134] 136. The method of any preceding statement, wherein the AXL-related disease is characterised by cells having a STK11 mutation and/or a STK11IP mutation.
[1135] 137. The method of any preceding statement, wherein the AXL-related disease is characterised by cells having a KRAS mutation.
[1136] 138. The method of any preceding statement, wherein the AXL-related disease is characterised by the presence of cells having a p53 mutation.
[1137] 139. The method of any preceding statement, wherein the AXL-related disease is characterised by the presence of cells having a STK11 mutation, a KRAS mutation, and a p53 mutation.
[1138] 140. The method of any preceding statement, wherein the AXL-related disease is characterised by the presence of cells having a STK11IP mutation, a KRAS mutation, and a p53 mutation.
[1139] 141. The method of any preceding statement, wherein the AXL-related disease is not an AXL-related disease characterised by increased KRAS activity or expression, and wild-type STK11 and/or p53 activity or expression.
[1140] 142. The method of any preceding statement, wherein the AXL-related disease is not an AXL-related disease characterised by: the presence of cells having a KRAS mutation; and, the absence of cells having a STK11, STK11IP, and/or p53 mutation.
[1141] 143. The method of any preceding statement, wherein the AXL-related disease is not an AXL-related disease characterised by the presence of cells having a KRAS G12C mutation.
[1142] 144. The method of any preceding statement, wherein the AXL-related disease is not an AXL-related disease characterised by cells which do not have a STK11, STK11IP, and/or p53 mutation.
[1143] 145. The method of any preceding statement, wherein the AXL-related disease is not a Lewis Lung Carcinoma (LLC) or Lewis Lung model tumour.
[1144] Treatment Step
[1145] 146. The method of any preceding statement, further comprising treating the subject in a method of treatment according to any one of statements 201-285.
[1146] 147. The method according to any one of statements 101-145, further comprising administering to the subject a therapeutically effective amount of an AXL inhibitor (AXLi), an immune checkpoint modulator (ICM), and/or a chemotherapeutic agent and/or radiotherapy.
[1147] 148. The method of statement 147, wherein the AXL inhibitor is administered concurrently with the immune checkpoint modulator (ICM).
[1148] 149. The method of statement 147, wherein the AXL inhibitor is administered concurrently with the immune checkpoint modulator (ICM) and/or the chemotherapeutic agent and/or radiotherapy.
[1149] 150. The method of statement 147, wherein the AXL inhibitor is administered separately and/or sequentially to the immune checkpoint modulator (ICM).
[1150] 151. The method of statement 147, wherein the AXL inhibitor is administered separately and/or sequentially to the immune checkpoint modulator (ICM) and/or the chemotherapeutic agent and/or radiotherapy.
[1151] 152. The method of statement 147, wherein the AXL inhibitor is administered subsequent to administration of the immune checkpoint modulator (ICM).
[1152] 153. The method of statement 147, wherein the AXL inhibitor is administered subsequent to administration of the immune checkpoint modulator (ICM) and/or subsequent to administration of the chemotherapeutic agent and/or radiotherapy.
[1153] 154. The method of statement 147, wherein the immune checkpoint modulator (ICM) is administered subsequent to administration of the Axl inhibitor.
[1154] 155. The method of statement 147, wherein the immune checkpoint modulator (ICM) is administered subsequent to administration of the Axl inhibitor and/or subsequent to administration of the chemotherapeutic agent and/or radiotherapy.
[1155] 156. The method of statement 147, wherein the chemotherapeutic agent and/or radiotherapy is administered subsequent to administration of the AXL inhibitor and/or subsequent to administration of the immune checkpoint modulator (ICM).
[1156] 157. The method of statement 147, wherein: [1157] i) the chemotherapeutic agent and/or radiotherapy is administered subsequent to administration of the AXL inhibitor; and [1158] ii) the immune checkpoint modulator (ICM) is administered subsequent to administration of the chemotherapeutic agent and/or radiotherapy.
[1159] 158. The method of statement 147, wherein the method comprises: [1160] i) administering the AXL inhibitor to the subject, wherein the immune checkpoint modulator (ICM) has been, is, or will be, administered to the subject; and/or [1161] ii) administering the AXL inhibitor to the subject, wherein the chemotherapeutic agent and/or radiotherapy has been, is, or will be, administered to the subject.
[1162] 159. The method of statement 147, wherein the method comprises: [1163] i) administering the immune checkpoint modulator (ICM) to the subject, wherein the AXL inhibitor has been, is, or will be, administered to the subject; and/or [1164] ii) administering the immune checkpoint modulator (ICM) to the subject, wherein the chemotherapeutic agent and/or radiotherapy has been, is, or will be, administered to the subject.
[1165] 160. The method of statement 147, wherein the method comprises: [1166] i) administering the chemotherapeutic agent and/or radiotherapy to the subject, wherein the AXL inhibitor has been, is, or will be, administered to the subject; and/or [1167] ii) administering the chemotherapeutic agent and/or radiotherapy to the subject, wherein the immune checkpoint modulator (ICM) has been, is, or will be, administered to the subject.
[1168] AXLi
[1169] 161. The method of any preceding statement, wherein the AXL inhibitor is a compound of formula (I) as set out in the description.
[1170] 162. The method of statement 161, wherein the AXL inhibitor is selected from the group consisting of: [1171] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1172] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7-(S)-pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1173] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7-(R)-pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1174] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(3-fluoro-4-(4-(pyrrolidin-1-yl)piperidin-1-yl)phenyl)-1H-1,2,4-triazole-3,5-diamine; [1175] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N5-(7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-1-yl)-1H-1,2,4-triazole-3,5-diamine; [1176] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N5-(7-(S)-pyrrolidin-1-yl-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1177] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(t-butoxycarbonylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1178] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(acetamido)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1179] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-((2R)-2-(methoxycarbonyl)pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1180] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4,4-difluoropiperidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1181] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-((methoxycarbonylmethyl)(methyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1182] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-((2R)-2-(carboxy)pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1183] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4-(ethoxycarbonyl)piperidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1184] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4-(carboxy)piperidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1185] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-((carboxymethyl)(methyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1186] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4-(ethoxycarbonylmethyl)piperazin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1187] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4-(carboxymethyl)piperazin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1188] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-1-yl)-1H-1,2,4-triazole-3,5-diamine; [1189] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1190] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7s)-7-(di(cyclopropylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1191] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((2-methylpropyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1192] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((propyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1193] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(dipropylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1194] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(diethylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1195] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclohexylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1196] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclopentylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1197] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((1-cyclopentylethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1198] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(2-propylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1199] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((3,3-dimethylbut-2-yl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1200] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((cyclohexylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1201] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(cyclohexylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1202] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((5-chlorothien-2-yl)methyl)amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1203] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((2-carboxyphenyl)methyl)amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1204] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((3-bromophenyl)methyl)amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1205] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(dimethylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1206] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclobutylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1207] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(3-pentylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1208] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((2,2-dimethylpropyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1209] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(cyclopentylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1210] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((cyclopentylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1211] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(bicyclo[2.2.1]hept-2-en-5-ylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1212] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((bicyclo[2.2.1]hept-2-en-5-ylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1213] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(3-methylbutylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1214] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(3-methylbutyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1215] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(2-ethylbutylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1216] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(but-2-enylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1217] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(butyl(but-2-enyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1218] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N5-((7S)-7-(t-butoxycarbonylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1219] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1220] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(dimethylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1221] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(diethylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1222] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(dipropylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1223] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(cyclopropylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1224] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(3-methylbutyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1225] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclobutylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1226] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclohexylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1227] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((methylethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1228] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclopentylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; and [1229] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(2-butylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1230] or pharmaceutically acceptable salts thereof.
[1231] 163. The method of statement 161, wherein the AXL inhibitor is 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7-(S)-pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine, or a pharmaceutically acceptable salt thereof.
[1232] 164. The method of statement 161, wherein the AXL inhibitor is Bemcentinib (BGB324/R428).
[1233] 165. The method of any one of statements 101-160, wherein the AXL inhibitor is selected from the group consisting of: dubermatinib (CAS No.1341200-45-0; UNII 14D65TV20J); gilteritinib (CAS No. 1254053-43-4; UNII 66D92MGC8M); cabozantinib (CAS No. 849217-68-1; UNII 1C39JW444G); SG17079 (CAS No. 1239875-86-5); merestinib (CAS No. 1206799-15-6; UNII 50GS5K699E); amuvatinib (CAS No. 850879-09-3; UNII SO9S6QZB4R); bosutinib (CAS No. 380843-75-4; UNII 5018V4AEZ0); glesatinib (CAS No. 936694-12-1; UNII 7Q290XD98N); foretinib (CAS No. 849217-64-7; UNII 81 FH7VK1 C4); and, TP0903 (CAS No. 1341200-45-0).
[1234] 166. The method of any one of statements 101-160, wherein the AXL inhibitor is an AXL inhibitor disclosed in WO2008/083367, WO2010/083465, or WO2012/028332.
[1235] 167. The method of any one of statements 101-160, wherein the AXL inhibitor is an anti-AXL antibody.
[1236] 168. The method of statement 167, wherein the antibody is an anti-AXL antibody disclosed in WO2015/193428, WO2015/193430, WO2016/097370, or WO2016/166296.
[1237] 169. The method of statement 167, wherein the antibody is an anti-AXL antibody selected from the group consisting of: the 1613F12 antibody disclosed in WO2013/064685; the 110D7 antibody disclosed in WO2014/068139; the 1003A2 antibody disclosed in WO2014/068139; the 1024G11 antibody disclosed in WO2014/068139; the hu10G5 antibody disclosed in WO2017/220695; and, the YW327.6S2 antibody disclosed in WO2011/159980.
[1238] 170. The method of statement 167, wherein the antibody comprises the 6 CDRs having the sequences of SEQ ID Nos. 1 to 6.
[1239] 171. The method of statement 167, wherein the antibody comprises the 6 CDRs having the sequences of SEQ ID Nos. 7 to 12.
[1240] 172. The method of statement 167, wherein the antibody comprises: [1241] a VH domain having the sequence of SEQ ID No. 13 and a VL domain having the sequence of SEQ ID NO. 15; [1242] a VH domain having the sequence of SEQ ID No. 13 and a VL domain having the sequence of SEQ ID NO. 16; [1243] a VH domain having the sequence of SEQ ID No. 14 and a VL domain having the sequence of SEQ ID NO. 15; or [1244] a VH domain having the sequence of SEQ ID No. 14 and a VL domain having the sequence of SEQ ID NO. 16.
[1245] ICM
[1246] 173. The method of any preceding statement, wherein the immune checkpoint modulator includes one or more immune checkpoint inhibitors (101).
[1247] 174. The method of any preceding statement, wherein the immune checkpoint modulator includes one or more immune checkpoint modulating antibody.
[1248] 175. The method of statement 174, wherein one or more immune checkpoint modulating antibody is selected from the group consisting of: anti-CTLA-4 antibodies, anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-4-1 BB antibodies, anti-OX-40 antibodies, anti-GITR antibodies, anti-CD27 antibodies, anti-CD28 antibodies, anti-CD40 antibodies, anti-LAGS antibodies, anti-ICOS antibodies, anti-TWEAKR antibodies, anti-HVEM antibodies, anti-TIM-1 antibodies, anti-TIM-3 antibodies, anti-VISTA antibodies, and anti-TIGIT antibodies.
[1249] 176. The method of statement 174, wherein one or more immune checkpoint modulating antibody is selected from the group consisting of: anti-CTLA-4 antibodies, anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-4-1 BB antibodies, anti-OX-40 antibodies, anti-GITR antibodies, anti-CD27 antibodies, anti-CD40 antibodies, and anti-LAGS antibodies.
[1250] 177. The method of statement 174, wherein one or more immune checkpoint modulating antibody is selected from the group consisting of: anti-CTLA-4 antibodies, anti-PD-1 antibodies, and anti-PD-L1 antibodies.
[1251] 178. The method of any preceding statement, wherein the immune checkpoint modulator includes: one or more T-cell co-stimulatory agonist; and/or one or more dendritic cell co-stimulatory receptor agonist.
[1252] 179. The method of any preceding statement, wherein the immune checkpoint modulator includes at least two immune checkpoint modulators.
[1253] 180. The method of any preceding statement, wherein the immune checkpoint modulator includes: (i) an immune checkpoint inhibitor, and (ii) a T cell co-stimulatory receptor agonist or a dendritic cell co-stimulatory receptor agonist.
[1254] 181. The method of any preceding statement, wherein the immune checkpoint modulator includes: (i) an anti-CTLA-4 antibody; and, (ii) an anti-PD-1 antibody and/or an anti-PD-L1 antibody.
[1255] 182. The method of statement 181, wherein the anti-CTLA-4 antibody is ipilimumab or tremelimumab.
[1256] 183. The method of statement 181, wherein the anti-PD-1 antibody is pembrolizumab or nivolumab.
[1257] 184. The method of statement 181, wherein the anti-PD-L1 antibody is atezolizumab (CAS number 1380723-44-3), avelumab (CAS number 1537032-82-8), or durvalumab (CAS number 1428935-60-7).
[1258] 185. The method of any preceding statement, wherein the immune checkpoint modulator includes, or is: pembrolizumab; ipilimumab; ipilimumab and nivolumab; ipilimumab and pembrolizumab; tremelilumab and durvalumab.
[1259] 186. The method of any one of statements 179 to 185, wherein the at least two immune checkpoint modulators are administered concurrently.
[1260] 187. The method of any one of statements 179 to 185, wherein the at least two immune checkpoint modulators are administered separately and/or sequentially.
[1261] Chemotherapeutic Agent/Radiotherapy
[1262] 188. The method of any one of statements 101-104 and 109-187, wherein the treatment comprises treatment with a combination of an AXL inhibitor (AXLi), an immune checkpoint modulator (ICM), and a chemotherapeutic agent and/or radiotherapy.
[1263] 189. The method of any one of statements 105-188, wherein the chemotherapeutic agent is a chemotherapeutic agent which induces immunogenic cell death of cancer cells.
[1264] 190. The method of any one of statements 105-188, wherein the chemotherapeutic agent is a chemotherapeutic agent which induces an immune response in the subject.
[1265] 191. The method of any one of statements 105-188, wherein the chemotherapeutic agent is a chemotherapeutic agent which induces a type I interferon response in the subject.
[1266] 192. The method of any one of statements 105-188, wherein the chemotherapeutic agent is an anthracycline.
[1267] 193. The method of statement 192, wherein the anthracycline is doxorubicin, daunorubicin, epirubicin, idarubicin, mitoxantrone, or valrubicin.
[1268] 194. The method of statement 192, wherein the anthracycline is doxorubicin.
[1269] Methods of Treatment
[1270] 201a. A method of treating an AXL-related disease in a subject with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: [1271] (i) administering a combination of the AXLi and the ICM to the subject; [1272] (ii) administering the AXLi to the subject, wherein the ICM has been, is, or will be, administered to the subject; or [1273] (iii) administering the ICM to the subject, wherein the AXLi has been, is, or will be, administered to the subject; [1274] wherein the AXL-related disease is characterised by the presence of cells having modified STK11 activity or expression.
[1275] 201b. A method of treating an AXL-related disease in a subject with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: [1276] (i) administering a combination of the AXLi and the ICM to the subject; [1277] (ii) administering the AXLi to the subject, wherein the ICM has been, is, or will be, administered to the subject; or [1278] (iii) administering the ICM to the subject, wherein the AXLi has been, is, or will be, administered to the subject; [1279] wherein the AXL-related disease is characterised by the presence of cells having increased KRAS activity or expression.
[1280] 202a. A method of treating an AXL-related disease in a subject with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: [1281] (i) administering a combination of the AXLi and the ICM to the subject; [1282] (ii) administering the AXLi to the subject, wherein the ICM has been, is, or will be, administered to the subject; or [1283] (iii) administering the ICM to the subject, wherein the AXLi has been, is, or will be, administered to the subject; [1284] wherein the AXL-related disease is characterised by the presence of cells having a STK11 mutation or a STK11IP mutation.
[1285] 202b. A method of treating an AXL-related disease in a subject with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: [1286] (i) administering a combination of the AXLi and the ICM to the subject; [1287] (ii) administering the AXLi to the subject, wherein the ICM has been, is, or will be, administered to the subject; or [1288] (iii) administering the ICM to the subject, wherein the AXLi has been, is, or will be, administered to the subject; [1289] wherein the AXL-related disease is characterised by the presence of cells having a KRAS mutation.
[1290] 203a. A method of treating an AXL-related disease in a subject with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: [1291] (i) administering a combination of the AXLi and the ICM to the subject; [1292] (ii) administering the AXLi to the subject, wherein the ICM has been, is, or will be, administered to the subject; or [1293] (iii) administering the ICM to the subject, wherein the AXLi has been, is, or will be, administered to the subject; [1294] wherein the subject has been selected for treatment on the basis that the AXL-related disease is characterised by the presence of cells having decreased STK11 activity or expression.
[1295] 203b. A method of treating an AXL-related disease in a subject with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: [1296] (i) administering a combination of the AXLi and the ICM to the subject; [1297] (ii) administering the AXLi to the subject, wherein the ICM has been, is, or will be, administered to the subject; or [1298] (iii) administering the ICM to the subject, wherein the AXLi has been, is, or will be, administered to the subject; [1299] wherein the subject has been selected for treatment on the basis that the AXL-related disease is characterised by the presence of cells having increased KRAS activity or expression.
[1300] 204. A method of treating an AXL-related disease in a subject with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: [1301] (i) administering a combination of the AXLi and the ICM to the subject; [1302] (ii) administering the AXLi to the subject, wherein the ICM has been, is, or will be, administered to the subject; or [1303] (iii) administering the ICM to the subject, wherein the AXLi has been, is, or will be, administered to the subject; [1304] wherein the subject has been selected for treatment using a method according to any one of statements 101-194.
Patient Sub-Group Definition
[1305] 202. The method of any preceding statement, wherein the AXL-related disease is further characterised by the presence of cells having increased KRAS activity or expression.
[1306] 203. The method of any preceding statement, wherein the AXL-related disease is further characterised by the presence of cells having decreased p53 activity or expression.
[1307] 204. The method of any preceding statement, wherein the AXL-related disease is further characterised by increased activity or expression of AXL.
[1308] 205. The method of any preceding statement, wherein the AXL-related disease is further characterised by the presence of cells having a KRAS mutation.
[1309] 206. The method of any preceding statement, wherein the AXL-related disease is further characterised by the presence of cells having a p53 mutation.
[1310] 207. The method of any preceding statement, wherein the AXL-related disease is characterised by the presence of cells having a STK11 mutation, a KRAS mutation, and a p53 mutation.
[1311] 208. The method of any preceding statement, wherein the AXL-related disease is characterised by the presence of cells having a STK11IP mutation, a KRAS mutation, and a p53 mutation.
[1312] 209. The method of any preceding statement, wherein the STK11 mutation, STK11IP mutation, KRAS mutation, and/or p53 mutation is a mutation selected from: [1313] (i) a mutation in the nucleotide sequence encoding STK11, STK11IP, KRAS, or p53; [1314] (ii) a mutation in a regulatory sequence controlling expression of the nucleotide sequence encoding STK11, STK11IP, KRAS, or p53; or [1315] (iii) a mutation in a nucleotide encoding a protein which interacts with the transcription product of the STK11, STK11IP, KRAS, or p53 gene.
[1316] 210. The method of any preceding statement, wherein the STK11 mutation, STK11IP mutation, KRAS mutation, and/or p53 mutation is a mutation in the translation product of the STK11, STK11IP, KRAS, or p53 gene.
[1317] 211. The method of any preceding statement, wherein the STK11 mutation, STK11IP mutation, KRAS mutation, and/or p53 mutation is a mutation in the transcription product of the STK11, STK11IP, KRAS, or p53 gene.
[1318] 212. The method of any preceding statement, wherein the STK11 mutation is an inactivating mutation.
[1319] 213. The method of any preceding statement, wherein the STK11IP mutation is an activating mutation.
[1320] 214. The method of any preceding statement, wherein the KRAS mutation is an activating mutation.
[1321] 215. The method of any preceding statement, wherein the p53 mutation is an inactivating mutation.
[1322] 216. The method of any preceding statement, wherein the STK11 mutation results in a reduced level of activity or expression of STK11 protein.
[1323] 217. The method of any preceding statement, wherein the STK11IP mutation results in an increased level of activity or expression of STK11IP protein.
[1324] 218. The method of any preceding statement, wherein the STK11IP mutation results in an altered pattern of activity or expression of STK11 protein, and/or altered subcellular localisation of STK11 protein.
[1325] 219. The method of any preceding statement, wherein the KRAS mutation results in an increased level of activity or expression of KRAS protein.
[1326] 220. The method of statement 219, wherein the KRAS mutation is a mutation at position G12, optionally wherein the KRAS mutation is a G12D mutation.
[1327] 221. The method of any preceding statement, wherein the p53 mutation results in a reduced level of activity or expression of p53 protein.
[1328] 222. The method of any preceding statement, wherein increased or decreased activity or expression is determined in a sample derived from a subject.
[1329] 223. The method of any preceding statement, wherein increased or decreased activity or expression is determined relative to a control.
[1330] 224. The method of statement 224, wherein the control is healthy tissue, preferably of the same tissue type as the AXL-related disease.
[1331] AXL-Related Disease
[1332] 225. The method of any preceding statement, wherein the AXL-related disease is a proliferative disease.
[1333] 226. The method of any preceding statement, wherein the AXL-related disease is a neoplastic disease.
[1334] 227. The method of any preceding statement, wherein the AXL-related disease is a solid tumour.
[1335] 228. The method of any preceding statement, wherein the AXL-related disease is cancer.
[1336] 229. The method of statement 228, wherein the cancer is selected from the group consisting of: lung cancer, non-small-cell lung cancer, breast cancer, melanoma, mesothelioma, acute myeloid leukemia (AML), myelodysplatic syndrome (MDS), pancreas cancer, kidney cancer, urothelial carcinoma, and glioblastoma.
[1337] 230. The method of statement 228, wherein the cancer is lung cancer.
[1338] 231. The method of statement 228, wherein the cancer is non-small-cell lung cancer (NSCLC). 232. The method of any preceding statement, wherein the AXL-related disease is not an AXL-related disease characterised by increased KRAS activity or expression, and wild-type STK11 and/or p53 activity or expression.
[1339] 233. The method of any preceding statement, wherein the AXL-related disease is not an AXL-related disease characterised by: the presence of cells having a KRAS mutation; and, the absence of cells having a STK11, STK11IP, and/or p53 mutation.
[1340] 234. The method of any preceding statement, wherein the AXL-related disease is not an AXL-related disease characterised by the presence of cells having a KRAS G12C mutation.
[1341] 235. The method of any preceding statement, wherein the AXL-related disease is not an AXL-related disease characterised by cells which do not have a STK11, STK11IP, and/or p53 mutation.
[1342] 236. The method of any preceding statement, wherein the AXL-related disease is not a Lewis Lung Carcinoma (LLC) or Lewis Lung model tumour.
[1343] 237. The method of any preceding statement, wherein the AXL-related disease does not respond to or benefit from treatment with an immune checkpoint modulator (ICM) when administered alone or as part of a treatment regime that does not include an AXLi.
[1344] AXLi
[1345] 238. The method of any preceding statement, wherein the AXL inhibitor is a compound of formula (I) as set out in the description.
[1346] 239. The method of statement 238, wherein the AXL inhibitor is selected from the group consisting of: [1347] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1348] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7-(S)-pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1349] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7-(R)-pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1350] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(3-fluoro-4-(4-(pyrrolidin-1-yl)piperidin-1-yl)phenyl)-1H-1,2,4-triazole-3,5-diamine;
[1351] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N5-(7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-1-yl)-1H-1,2,4-triazole-3,5-diamine; [1352] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N5-(7-(S)-pyrrolidin-1-yl-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1353] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(t-butoxycarbonylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1354] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(acetamido)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1355] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-((2R)-2-(methoxycarbonyl)pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1356] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4,4-difluoropiperidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1357] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-((methoxycarbonylmethyl)(methyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1358] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-((2R)-2-(carboxy)pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1359] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4-(ethoxycarbonyl)piperidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1360] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4-(carboxy)pipendin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1361] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-((carboxymethyl)(methyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1362] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4-(ethoxycarbonylmethyl)piperazin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1363] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(4-(carboxymethyl)piperazin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1364] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-(7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-1-yl)-1H-1,2,4-triazole-3,5-diamine; [1365] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1366] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7s)-7-(di(cyclopropylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1367] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((2-methylpropyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1368] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((propyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1369] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(dipropylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1370] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(diethylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1371] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclohexylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1372] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclopentylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1373] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((1-cyclopentylethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1374] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(2-propylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1375] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((3,3-dimethylbut-2-Aamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1376] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((cyclohexylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1377] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(cyclohexylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1378] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((5-chlorothien-2-yl)methyl)amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1379] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((2-carboxyphenyl)methyl)amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1380] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((3-bromophenyl)methyl)amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1381] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(dimethylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1382] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclobutylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1383] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(3-pentylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1384] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((2,2-dimethylpropyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1385] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(cyclopentylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1386] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((cyclopentylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1387] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(bicyclo[2.2.1]hept-2-en-5-ylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1388] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)((bicyclo[2.2.1]hept-2-en-5-ylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1389] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(3-methylbutylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1390] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(3-methylbutyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1391] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(2-ethylbutylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1392] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(but-2-enylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine;
[1393] 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(butyl(but-2-enyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1394] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N5-((7S)-7-(t-butoxycarbonylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1395] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-amino-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1396] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(dimethylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1397] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(diethylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1398] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(dipropylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1399] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(cyclopropylmethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1400] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(di(3-methylbutyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1401] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclobutylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1402] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclohexylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1403] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-((methylethyl)amino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1404] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(cyclopentylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; and [1405] 1-(6,7-dihydro-5H-pyrido[2′,3′:6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7S)-7-(2-butylamino)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine; [1406] or pharmaceutically acceptable salts thereof.
[1407] 240. The method of statement 238, wherein the AXL inhibitor is 1-(6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazin-3-yl)-N3-((7-(S)-pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine, or a pharmaceutically acceptable salt thereof.
[1408] 241. The method of statement 238, wherein the AXL inhibitor is Bemcentinib (BGB324/R428).
[1409] 242. The method of any one of statements 201-237, wherein the AXL inhibitor is selected from the group consisting of: dubermatinib (CAS No. 1341200-45-0; UNII 14D65TV20J); gilteritinib (CAS No. 1254053-43-4; UNII 66D92MGC8M); cabozantinib (CAS No. 849217-68-1; UNII 1C39JW444G); SG17079 (CAS No. 1239875-86-5); merestinib (CAS No. 1206799-15-6; UNII 50GS5K699E); amuvatinib (CAS No. 850879-09-3; UNII SO9S6QZB4R); bosutinib (CAS No. 380843-75-4; UNII 5018V4AEZ0); glesatinib (CAS No. 936694-12-1; UNII 7Q290XD98N); foretinib (CAS No. 849217-64-7; UNII 81 FH7VK1 C4); and, TP0903 (CAS No. 1341200-45-0).
[1410] 243. The method of any one of statements 201-237, wherein the AXL inhibitor is an AXL inhibitor disclosed in WO2008/083367, WO2010/083465, or WO2012/028332.
[1411] 244. The method of any one of statements 201-237, wherein the AXL inhibitor is an anti-AXL antibody.
[1412] 245. The method of statement 244, wherein the antibody is an anti-AXL antibody disclosed in WO2015/193428, WO2015/193430, WO2016/097370, or WO2016/166296.
[1413] 246. The method of statement 244, wherein the antibody is an anti-AXL antibody selected from the group consisting of: the 1613F12 antibody disclosed in W02013/064685; the 110D7 antibody disclosed in WO2014/068139; the 1003A2 antibody disclosed in WO2014/068139; the 1024G11 antibody disclosed in WO2014/068139; the hu10G5 antibody disclosed in WO2017/220695; and, the YW327.6S2 antibody disclosed in WO2011/159980.
[1414] 247. The method of statement 244, wherein the antibody comprises the 6 CDRs having the sequences of SEQ ID Nos. 1 to 6.
[1415] 248. The method of statement 244, wherein the antibody comprises the 6 CDRs having the sequences of SEQ ID Nos. 7 to 12.
[1416] 249. The method of statement 244, wherein the antibody comprises: [1417] a VH domain having the sequence of SEQ ID No. 13 and a VL domain having the sequence of SEQ ID NO.15; [1418] a VH domain having the sequence of SEQ ID No. 13 and a VL domain having the sequence of SEQ ID NO.16; [1419] a VH domain having the sequence of SEQ ID No. 14 and a VL domain having the sequence of SEQ ID NO.15; or [1420] a VH domain having the sequence of SEQ ID No. 14 and a VL domain having the sequence of SEQ ID NO.16.
[1421] ICM
[1422] 250. The method of any preceding statement, wherein the immune checkpoint modulator includes one or more immune checkpoint inhibitors (101).
[1423] 251. The method of any preceding statement, wherein the immune checkpoint modulator includes one or more immune checkpoint modulating antibody.
[1424] 252. The method of statement 251, wherein one or more immune checkpoint modulating antibody is selected from the group consisting of: anti-CTLA-4 antibodies, anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-4-1 BB antibodies, anti-OX-40 antibodies, anti-GITR antibodies, anti-CD27 antibodies, anti-CD28 antibodies, anti-CD40 antibodies, anti-LAGS antibodies, anti-ICOS antibodies, anti-TWEAKR antibodies, anti-HVEM antibodies, anti-TIM-1 antibodies, anti-TIM-3 antibodies, anti-VISTA antibodies, and anti-TIGIT antibodies.
[1425] 253. The method of statement 251, wherein one or more immune checkpoint modulating antibody is selected from the group consisting of: anti-CTLA-4 antibodies, anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-4-1 BB antibodies, anti-OX-40 antibodies, anti-GITR antibodies, anti-CD27 antibodies, anti-CD40 antibodies, and anti-LAGS antibodies.
[1426] 254. The method of statement 251, wherein one or more immune checkpoint modulating antibody is selected from the group consisting of: anti-CTLA-4 antibodies, anti-PD-1 antibodies, and anti-PD-L1 antibodies.
[1427] 255. The method of any preceding statement, wherein the immune checkpoint modulator includes: one or more T-cell co-stimulatory agonist; and/or one or more dendritic cell co-stimulatory receptor agonist.
[1428] 256. The method of any preceding statement, wherein the immune checkpoint modulator includes at least two immune checkpoint modulators.
[1429] 257. The method of any preceding statement, wherein the immune checkpoint modulator includes: (i) an immune checkpoint inhibitor, and (ii) a T cell co-stimulatory receptor agonist or a dendritic cell co-stimulatory receptor agonist.
[1430] 258. The method of any preceding statement, wherein the immune checkpoint modulator includes: (i) an anti-CTLA-4 antibody; and, (ii) an anti-PD-1 antibody and/or an anti-PD-L1 antibody.
[1431] 259. The method of statement 258, wherein the anti-CTLA-4 antibody is ipilimumab or tremelimumab.
[1432] 260. The method of statement 258, wherein the anti-PD-1 antibody is pembrolizumab or nivolumab.
[1433] 261. The method of statement 258, wherein the anti-PD-L1 antibody is atezolizumab (CAS number 1380723-44-3), avelumab (CAS number 1537032-82-8), or durvalumab (CAS number 1428935-60-7).
[1434] 262. The method of any preceding statement, wherein the immune checkpoint modulator includes, or is: pembrolizumab; ipilimumab; ipilimumab and nivolumab; ipilimumab and pembrolizumab; tremelilumab and durvalumab.
[1435] 263. The method of any one of statements 256 to 262, wherein the at least two immune checkpoint modulators are administered concurrently.
[1436] 264. The method of any one of statements 256 to 262, wherein the at least two immune checkpoint modulators are administered separately and/or sequentially.
[1437] Triple Combination with Chemotherapeutic Agent/Radiotherapy
[1438] 265. The method of any preceding statement, wherein the subject is treated with a combination of an AXL inhibitor (AXLi), an immune checkpoint modulator (ICM), and a chemotherapeutic agent and/or radiotherapy.
[1439] 266. The method of statement 265, wherein the chemotherapeutic agent is a chemotherapeutic agent which induces immunogenic cell death of cancer cells.
[1440] 267. The method of any one of statements 265-266, wherein the chemotherapeutic agent is a chemotherapeutic agent which induces an immune response in the subject.
[1441] 268. The method of any one of statements 265-267, wherein the chemotherapeutic agent is a chemotherapeutic agent which induces a type I interferon response in the subject.
[1442] 269. The method of any one of statements 265-268, wherein the chemotherapeutic agent is an anthracycline.
[1443] 270. The method of statement 269, wherein the anthracycline is doxorubicin, daunorubicin, epirubicin, idarubicin, mitoxantrone, or valrubicin.
[1444] 271. The method of statement 269, wherein the anthracycline is doxorubicin.
[1445] 272. The method of any one of statements 265-271, wherein treatment of the AXL-related disease with a combination of immune checkpoint modulator (ICM) and chemotherapeutic agent does not provide any additional benefit as compared to treatment with the chemotherapeutic agent alone.
[1446] Administration Schedule Features
[1447] 273. The method of any one of statements 201-272, wherein the AXL inhibitor is administered concurrently with the immune checkpoint modulator (ICM).
[1448] 274. The method of any one of statements 265-272, wherein the AXL inhibitor is administered concurrently with the immune checkpoint modulator (ICM) and/or the chemotherapeutic agent and/or radiotherapy.
[1449] 275. The method of any one of statements 201-272, wherein the AXL inhibitor is administered separately and/or sequentially to the immune checkpoint modulator (ICM).
[1450] 276. The method of any one of statements 265-272, wherein the AXL inhibitor is administered separately and/or sequentially to the immune checkpoint modulator (ICM) and/or the chemotherapeutic agent and/or radiotherapy.
[1451] 277. The method of any one of statements 201-272, wherein the AXL inhibitor is administered subsequent to administration of the immune checkpoint modulator (ICM).
[1452] 278. The method of any one of statements 265-272, wherein the AXL inhibitor is administered subsequent to administration of the immune checkpoint modulator (ICM) and/or subsequent to administration of the chemotherapeutic agent and/or radiotherapy.
[1453] 279. The method of any one of statements 201-272, wherein the immune checkpoint modulator (ICM) is administered subsequent to administration of the Axl inhibitor.
[1454] 280. The method of any one of statements 265-272, wherein the immune checkpoint modulator (ICM) is administered subsequent to administration of the Axl inhibitor and/or subsequent to administration of the chemotherapeutic agent and/or radiotherapy.
[1455] 281. The method of any one of statements 265-272, wherein the chemotherapeutic agent and/or radiotherapy is administered subsequent to administration of the AXL inhibitor and/or subsequent to administration of the immune checkpoint modulator (ICM).
[1456] 282. The method of any one of statements 265-272, wherein: [1457] the chemotherapeutic agent and/or radiotherapy is administered subsequent to administration of the AXL inhibitor; and [1458] the immune checkpoint modulator (ICM) is administered subsequent to administration of the chemotherapeutic agent and/or radiotherapy.
[1459] 283. The method of any one of statements 265-272, wherein the method comprises: [1460] i) administering the AXL inhibitor to the subject, wherein the immune checkpoint modulator (ICM) has been, is, or will be, administered to the subject; and/or [1461] ii) administering the AXL inhibitor to the subject, wherein the chemotherapeutic agent and/or radiotherapy has been, is, or will be, administered to the subject.
[1462] 284. The method of any one of statements 265-272, wherein the method comprises: [1463] i) administering the immune checkpoint modulator (ICM) to the subject, wherein the AXL inhibitor has been, is, or will be, administered to the subject; and/or [1464] ii) administering the immune checkpoint modulator (ICM) to the subject, wherein the chemotherapeutic agent and/or radiotherapy has been, is, or will be, administered to the subject.
[1465] 285. The method of any one of statements 265-272, wherein the method comprises: [1466] i) administering the chemotherapeutic agent and/or radiotherapy to the subject, wherein the AXL inhibitor has been, is, or will be, administered to the subject; and/or [1467] ii) administering the chemotherapeutic agent and/or radiotherapy to the subject, wherein the immune checkpoint modulator (ICM) has been, is, or will be, administered to the subject.
[1468] Second Medical Uses
[1469] 301. An AXL inhibitor for use in a method of treating an AXL-related disease according to any one of statements 201-285.
[1470] 302. An immune checkpoint modulator (ICM) for use in a method of treating an AXL-related disease according to any one of statements 201-285.
[1471] 303. An AXL inhibitor and an immune checkpoint modulator (ICM) for use in a method of treating an AXL-related disease according to any one of statements 201-285.
[1472] 304. A chemotherapeutic agent for use in a method of treating an AXL-related disease according to any one of statements 265-285.
[1473] 305. An AXL inhibitor, an immune checkpoint modulator (1CM), and a chemotherapeutic agent for use in a method of treating an AXL-related disease according to any one of statements 265-285.
[1474] 306. A reagent for detecting activity, expression, or amount of STK11, STK11IP, KRAS, or p53, for use in a method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (1CM).
[1475] 307. A kit comprising 1, 2, 3, 4, or more reagents for detecting activity, expression, or amount of one or more of STK11, STK11IP, KRAS, or p53, for use in a method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM).
[1476] 308. The reagent or kit for use according to statement 306 or 307, wherein the method of selecting is a method according to any one of statements 101-194.
[1477] 309. The reagent or kit for use according to any one of statements 306-308, wherein each reagent for detecting is a specific binding member which is selective for STK11, STK11IP, KRAS, or p53.
[1478] 310. The reagent or kit for use according to any one of statements 306-308, wherein the reagent for detecting is an antibody, a nucleic acid probe, or a QPCR primer.
[1479] Swiss Format
[1480] 401. Use of an AXL inhibitor in the manufacture of a medicament for treating a disorder in a subject, wherein the treatment comprises a method of treating an AXL-related disease according to any one of statements 201-285.
[1481] 402. Use of an immune checkpoint modulator (ICM) in the manufacture of a medicament for treating a disorder in a subject, wherein the treatment comprises a method of treating an AXL-related disease according to any one of statements 201-285.
[1482] 403. Use of an AXL inhibitor and an immune checkpoint modulator (ICM) in the manufacture of a medicament for treating a disorder in a subject, wherein the treatment comprises a method of treating an AXL-related disease according to any one of statements 201-285.
[1483] 404. Use of a chemotherapeutic agent in the manufacture of a medicament for treating a disorder in a subject, wherein the treatment comprises a method of treating an AXL-related disease according to any one of statements 265-285.
[1484] 405. Use of an AXL inhibitor, an immune checkpoint modulator (ICM), and a chemotherapeutic agent in the manufacture of a medicament for treating a disorder in a subject, wherein the treatment comprises a method of treating an AXL-related disease according to any one of statements 265-285.
[1485] 406. Use of a reagent for detecting activity, expression, or amount of STK11, STK11IP, KRAS, or p53, in the manufacture of a kit or test for use in a method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM).
[1486] 407. Use of 1, 2, 3, 4, or more reagents for detecting activity, expression, or amount of one or more of STK11, STK11IP, KRAS, or p53, in the manufacture of a kit for use in a method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM).
[1487] 408. The use according to statement 406 or 407, wherein the method of selecting is a method according to any one of statements 101-194.
[1488] 409. The use according to any one of statements 406-408, wherein each reagent for detecting is a specific binding member which is selective for STK11, STK11IP, KRAS, or p53.
[1489] 410. The use according to any one of statements 406-408, wherein the reagent for detecting is an antibody, a nucleic acid probe, or a QPCR primer.
[1490] Kits
[1491] 501. A kit comprising an AXL inhibitor and an immune checkpoint modulator (ICM), for use in a method of treating an Axl-related disease according to any one of statements 201-285.
[1492] 502. A kit comprising an AXL inhibitor, an immune checkpoint modulator (ICM), and a chemotherapeutic agent, for use in a method of treating an Axl-related disease according to any one of statements 265-285.
[1493] 503. A kit comprising a chemotherapeutic agent and an AXL inhibitor and/or an immune checkpoint modulator (ICM), for use in a method of treating an Axl-related disease according to any one of statements 265-285.
[1494] Prognostic Methods
[1495] 601a. A method of prognosing susceptibility of a subject to treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: [1496] determining: i) the presence or absence of a STK11 mutation; and/or ii) the level of STK11 activity or expression in the subject or a sample derived from the subject; [1497] wherein the presence of a STK11 mutation and/or a modified level of STK11 activity or expression is indicative of susceptibility to treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM).
[1498] 601b. A method of prognosing susceptibility of a subject to treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: [1499] determining: i) the presence or absence of a KRAS mutation; and/or ii) the level of KRAS activity or expression in the subject or a sample derived from the subject; [1500] wherein the presence of a KRAS mutation and/or an increased level of KRAS activity or expression is indicative of susceptibility to treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM).
[1501] 602a. A method of prognosing susceptibility of a subject to treatment in a method according to any one of statements 201-285, the method comprising: [1502] determining: i) the presence or absence of a STK11 mutation; and/or ii) the level of STK11 activity or expression in the subject or a sample derived from the subject; [1503] wherein the presence of a STK11 mutation and/or a modified level of STK11 activity or expression is indicative of susceptibility to treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM).
[1504] 602b. A method of prognosing susceptibility of a subject to treatment in a method according to any one of statements 201-285, the method comprising: [1505] determining: i) the presence or absence of a KRAS mutation; and/or ii) the level of KRAS activity or expression in the subject or a sample derived from the subject; [1506] wherein the presence of a KRAS mutation and/or an increased level of KRAS activity or expression is indicative of susceptibility to treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM).
[1507] 603. The method according to any preceding statement, wherein the method further comprises: [1508] determining: i) the presence or absence of a KRAS mutation; and/or ii) the level of KRAS activity or expression in the subject or a sample derived from the subject; [1509] wherein the presence of a KRAS mutation and/or an increased level of KRAS activity or expression is indicative of susceptibility to treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM).
[1510] 604. The method of any preceding statement, wherein the method further comprises: [1511] determining: i) the presence or absence of a p53 mutation; and/or ii) the level of p53 activity or expression in the subject or a sample derived from the subject; [1512] wherein the presence of a p53 mutation and/or an decreased level of p53 activity or expression is indicative of susceptibility to treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM).
[1513] 605. The method of any preceding statement, wherein the method comprises: [1514] determining the level of AXL activity or expression in the subject or a sample derived from the subject; [1515] wherein an increased level of AXL activity or expression is indicative of susceptibility to treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM).
[1516] 606. A method of prognosing susceptibility of a subject to treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising testing subjects in a method according to any one of statements 101-194.
[1517] 607. A method of prognosing susceptibility of a subject to treatment in a method according to any one of statements 201-285, the method comprising testing subjects in a method according to any one of statements 101-194.
[1518] 608. The method of any one of statements 601-607, further comprising treating a subject determined to be susceptible to treatment in a method according to any one of statements 201-285.
[1519] 700. A method of selecting a subject for treatment with a combination of an AXL inhibitor (AXLi) and an immune checkpoint modulator (ICM), the method comprising: identifying subjects having an AXL-related disease characterised by a reduced presence of CD8.sup.+ cells having TCF1 activity or expression; and, selecting thus identified subjects for treatment.
[1520] 701. The method of statement 700, wherein the method further comprises: identifying subjects that have previously been treated with an immune checkpoint modulator (ICM) and which did not respond to treatment with the ICM; and, selecting thus identified subjects for treatment.
[1521] 702. The method of statement 700 or 701, further comprising the method of any one of statements 109-194.
[1522] 800. A method of increasing a population of desired T cells in a subject comprising treating the subject with an AXL inhibitor (AXLi).
[1523] 801. The method of statement 800, wherein the desired T cell population is a CD8+ T cell population.
[1524] 802. The method of 800 or 801, wherein the desired T cell population is a TCF1.sup.+PD-1.sup.+CD8.sup.+ T cell population.
[1525] 803. The method of any one of statements 800 to 802, wherein the subject is further treated with an immune checkpoint inhibitor as described in any one of statements 250-264.
REFERENCES
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