Treatment method of malignant peripheral nerve sheath tumor (MPNST) using LSD1 inhibitor

Abstract

The present invention relates to an LSD1 inhibitor for use in the treatment of malignant peripheral nerve sheath tumors (MPNSTs). The present invention also provides a method for treating MPNSTs in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of an LSD1 inhibitor.

Description

Technical Field

[0001] The present invention relates to the field of treating malignant peripheral nerve sheath tumors (MPNSTs). In particular, the present invention provides LSD1 inhibitors for use in treating MPNSTs. The present invention also provides a method for treating MPNSTs in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of an LSD1 inhibitor.

Background Art

[0002] Cancer is one of the leading causes of death worldwide. The clinical management of cancer has changed over the past few decades. Conventionally, cancer could only be treated with chemotherapeutic agents (less selective "dirty" drugs) that non-specifically (indiscriminately) limit cell proliferation by interfering with DNA and basic cell cycle mechanisms. Today, instead, a new approach of personalized precision medicine has been devised that blocks the growth of cancer cells with little damage to other cells in the body. The advantage of this approach is that targeted therapies are less harmful to normal cells.

[0003] Epigenetics is one of the emerging fields in cancer precision medicine, with first-generation drugs reaching the FDA approval and many more drugs in clinical trials. Lysine-specific demethylase 1 (LSD1 or KDM1A) is an epigenetic enzyme that regulates gene expression by demethylating two different residues of the histone H3 tail, namely lysine 4 (H3K4) and lysine 9 (H3K9), and has opposing effects. Demethylation of H3K4 is associated with transcriptional repression, while demethylation of H3K9 is associated with transcriptional activation. Furthermore, LSD1 is part of many multi-protein complexes that control enhancer-promoter contacts involved in gene repression, such as NurD and CoRest. LSD1 has been shown to play a major role in cancers such as leukemia and small cell lung cancer (SCLC), and significant efforts have been invested in the development of LSD1 inhibitors. Inhibitors targeting the catalytic active site, such as iadademstat, bomedemstat, and pulrodemstat, have been developed. These compounds bind deeply to the active site of LSD1 and inhibit both the catalytic activity and scaffolding interactions of LSD1 by blocking access to both the protein substrate (e.g., histone H3 tail) and non-substrate protein interactors (e.g., SNAG domain transcription factors). LSD1 inhibitors have been reported to have very potent anti-proliferative activity against specific tumor types and are currently in clinical trials as therapeutic agents for cancers such as acute myeloid leukemia (AML) and SCLC.

[0004] Malignant peripheral nerve sheath tumors (MPNSTs) are highly aggressive soft tissue sarcomas that can arise in at least three different contexts. Approximately 40-50% of MPNST cases occur in patients with neurofibromatosis type 1, an autosomal dominant genetic syndrome caused by mutations in the NF1 gene. Patients have a predisposition to learning disabilities and skeletal disorders and also tend to develop small benign tumors (neurofibromas). However, mutations in the NF1 gene alone are not sufficient to cause MPNSTs in patients with neurofibromatosis type 1, and a series of additional mutational events such as loss-of-function mutations in tumor suppressor genes other than NF1 (e.g., CDKN2A, p53, or PTEN) and / or mutations or copy number variations in oncogenes (e.g., EGFR or c-Met) need to accumulate. Furthermore, loss of polycomb repressive complex 2 (PRC2) activity is also often present. This multi-protein complex is involved in, among other processes, the methylation of histone H3 lysine 27 (H3K27), a known epigenetic post-translational modification particularly associated with transcriptional repression. Approximately 40% of MPNSTs are sporadic, and the genetic cause is unknown. The remaining approximately 10% occur as a result of previous radiotherapy. Generally, MPNSTs follow an aggressive clinical course, with a recurrence rate of 50% and an overall average 5-year survival rate of only 20-40%. Advanced disease often results in lung and bone metastases. Currently, there are no drugs specifically approved for the treatment of MPNSTs. Various drugs have been tested in clinical trials for MPNSTs, but these drugs have shown only disappointing response rates, and at best, only a small number of MPNST patients have achieved stable disease (see, for example, Martin E et al., Crit Rev Oncol Hematol, 2019, 138:223-32, doi:10.1016 / j.critrevonc.2019.04.007). Thus, there is a large unmet medical need for new and more effective treatments for MPNSTs. The present invention addresses this need and others.

Prior Art Documents

Non-Patent Documents

[0005]

Non-Patent Document 1

[0006] The present invention is based on the surprising discovery that LSD1 inhibitors such as iadademstat, bomedemstat, and pulrodemstat are beneficially effective in the treatment of MPNST, as also described in Example 1 below. This was completely unexpected, especially since MPNST is a very aggressive and extremely difficult-to-treat malignancy. Accordingly, the present invention provides a novel and improved therapeutic approach for the treatment of MPNST.

[0007] Accordingly, the present invention relates to an LSD1 inhibitor for use in the treatment of MPNST. The present invention also relates to a pharmaceutical composition comprising an LSD1 inhibitor and optionally one or more pharmaceutically acceptable excipients for use in the treatment of MPNST.

[0008] The present invention similarly provides a method of treating MPNST in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of an LSD1 inhibitor (or a therapeutically effective amount of a pharmaceutical composition comprising an LSD1 inhibitor and optionally one or more pharmaceutically acceptable excipients).

[0009] Furthermore, the present invention relates to the use of an LSD1 inhibitor for the treatment of MPNST. The present invention further relates to the use of an LSD1 inhibitor for the manufacture of a medicament (or pharmaceutical composition) for the treatment of MPNST. BRIEF DESCRIPTION OF THE DRAWINGS

[0010]

Figure 1

Figure 2

Mode for Carrying Out the Invention

[0011] As described above, the present invention is based on the surprising discovery that LSD1 inhibitors are beneficially effective in the treatment of MPNST, as will be further detailed in the following and the Examples section. In particular, the extensive panel of 12 different MPNST cell lines tested in Example 1 includes both neurofibromatosis type I-associated MPNST and sporadic MPNST cell lines, and is representative of the various subtypes of MPNST encountered in clinical practice, as it is characterized by significant genomic heterogeneity (variation) with respect to the heterozygosity of NF1, ploidy, and the mutation status of CDKN2A and PRC2. The exemplary LSD1 inhibitor, iadademstat, was found to be highly effective in a very diverse range of MPNST cell lines and is thus particularly well-suited for the treatment of MPNST. Additional LSD1 inhibitors with different chemical scaffolds, including both reversible and irreversible inhibitors of LSD1, were also confirmed to be effective in the treatment of MPNST, as described in Example 1 as well.

[0012] Accordingly, the present invention relates to an LSD1 inhibitor for use in the treatment of MPNST. The present invention also relates to a pharmaceutical composition for use in the treatment of MPNST, comprising an LSD1 inhibitor and optionally one or more pharmaceutically acceptable excipients. The present invention likewise provides a method for treating MPNST in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of an LSD1 inhibitor (or a therapeutically effective amount of a pharmaceutical composition comprising an LSD1 inhibitor and optionally one or more pharmaceutically acceptable excipients). Further, the present invention relates to the use of an LSD1 inhibitor for the treatment of MPNST. The present invention further relates to the use of an LSD1 inhibitor for the manufacture of a medicament (or pharmaceutical composition) for the treatment of MPNST.

[0013] According to the present invention, an "LSD1 inhibitor" refers to a compound that reduces, decreases, blocks or inhibits the gene expression, activity or function of LSD1. Examples thereof are provided below under the heading "LSD1 inhibitor". Suitable LSD1 inhibitors include each of iadademstat or a pharmaceutically acceptable salt thereof (e.g., iadademstat dihydrochloride), pruvostadstat or a pharmaceutically acceptable salt thereof (e.g., pruvostadstat besilate), and bomedemstat or a pharmaceutically acceptable salt thereof (e.g., bomedemstat bis tosylate). Particularly suitable LSD1 inhibitor is iadademstat or a pharmaceutically acceptable salt thereof (e.g., iadademstat dihydrochloride).

[0014] The LSD1 inhibitor (e.g., iadademstat or a pharmaceutically acceptable salt thereof) is preferably administered orally. Exemplary formulations that can be administered via oral administration, particularly oral ingestion, are described in more detail below.

[0015] The subject to be treated according to the present invention can be a human or an animal (e.g., a non-human mammal), preferably a human. The MPNST to be treated according to the present invention can be, for example, a neurofibromatosis type I-related MPNST, a sporadic MPNST, or a radiation-induced MPNST.

[0016] As described above, MPNST often occurs in subjects with neurofibromatosis type I, a hereditary disorder. Thus, in some embodiments, the MPNSTs treated according to the present invention are neurofibromatosis type I - associated MPNSTs (or MPNSTs associated with neurofibromatosis type I). Accordingly, in some embodiments, the present invention relates to the treatment of MPNSTs in subjects with neurofibromatosis type I. The present invention also relates to LSD1 inhibitors (or pharmaceutical compositions comprising LSD1 inhibitors) for use in the treatment of MPNSTs, wherein the LSD1 inhibitor (or its pharmaceutical composition) is administered to a subject with neurofibromatosis type I.

[0017] In some embodiments, the MPNSTs treated according to the present invention are MPNSTs not associated with neurofibromatosis type I (or MPNSTs not related to neurofibromatosis type I), particularly sporadic MPNSTs or radiation - induced MPNSTs. Accordingly, the present invention specifically relates to the treatment of MPNSTs in subjects not affected by neurofibromatosis type I. The present invention also relates to LSD1 inhibitors (or pharmaceutical compositions comprising LSD1 inhibitors) for use in the treatment of MPNSTs, wherein the LSD1 inhibitor (or its pharmaceutical composition) is administered to a subject not affected by neurofibromatosis type I. In some embodiments, the MPNST being treated is a sporadic MPNST. In some embodiments, the MPNST being treated is a radiation - induced MPNST (see, for example, Yamanaka R et al., World Neurosurg, 2017, 105:961 - 970.e8, doi:10.1016 / j.wneu.2017.06.010).

[0018] MPNSTs treated according to the present invention may further be MPNSTs having one or more mutations or genetic changes that affect the NF1 gene, particularly MPNSTs having one or more inactivating mutations or inactivating genetic changes in the NF1 gene (e.g., including any one of the above-identified types of MPNSTs). Thus, the MPNSTs to be treated can be type I neurofibromatosis-related MPNSTs having one or more mutations or genetic changes (e.g., one or more inactivating mutations or inactivating genetic changes) in the NF1 gene, sporadic MPNSTs having one or more mutations or genetic changes (e.g., one or more inactivating mutations or inactivating genetic changes) in the NF1 gene, or radiation-induced MPNSTs having one or more mutations or genetic changes (e.g., one or more inactivating mutations or inactivating genetic changes) in the NF1 gene. Such inactivating mutations or inactivating genetic changes in the NF1 gene particularly include loss-of-function mutations, resulting in a decrease or absence of the expression and / or stability and / or activity of its protein product neurofibromin. Further, such mutations or genetic changes can affect one or both alleles of the NF1 gene.

[0019] In humans, the NF1 gene is located on chromosome 17q11.2 and encodes a protein product called neurofibromin. The canonical (standard) amino acid sequence of isoform 2 of human neurofibromin is 2,839 residues long (see, for example, Uniprot identifier P21359-1; https: / / www.uniprot.org / uniprot / P21359.fasta), and that of isoform 1 is 2,818 residues long (see, for example, Uniprot identifier P21359-2; https: / / www.uniprot.org / uniprot / P21359-2.fasta). These two isoforms are thought to be the most biologically relevant. Isoform 2 is found in most human tissues but not in neurons of the central nervous system. The NF1 gene can generate other alternative splicing isoforms through various combinations of its approximately 60 exons. More than 3,000 germline mutations of the NF1 gene have been reported in the Human Gene Mutation Database (HGMD; http: / / www.hgmd.cf.ac.uk / ac / index.php), and more than 1,000 somatic mutations have been reported in The Cancer Genome Atlas (TCGA; https: / / www.cancer.gov / about-nci / organization / ccg / research / structural-genomics / tcga). See also Scheer M et al., Int J Mol Sci, 2021, 23(1):352, doi:10.3390 / ijms23010352. Many of the characterized mutations are loss-of-function, meaning that they ultimately have an adverse effect on the function of the protein product. At the gene level, all types of mutations have been reported, including nonsense, missense, frameshift, indels (insertions or deletions), microdeletions, inversions, splice site variants, whole translocations, and complex rearrangements. However, there is no clear pattern of local mutational clustering within the NF1 gene.To identify mutations or genetic changes in the NF1 gene, it is recommended to apply a mutation detection pipeline to reliably characterize NF1 mutants. There, a series of different algorithms specifically fine-tuned to detect single nucleotide variants (SNVs), indels or translocations in next-generation sequencing (NGS) reads are used. The presence of mutations or genetic changes in the NF1 gene can be evaluated in a sample obtained from a subject, such as a biopsy sample. The domain structure of neurofibromin is complex and, from the N-terminus to the C-terminus, includes: (1) an N-heat domain containing, among other things, a cysteine- and serine-rich domain / GTPase-activating domain (CSRD) and a tubulin-binding domain (TBD); (2) a GTPase-activating domain (GAP)-related domain (GRD) that promotes the hydrolysis of active Ras-GTP to inactive Ras-GDP; (3) a Sec14 homology segment; (4) a Pleckstrin homology (PH)-like domain; and (5) a C-heat domain containing a heat-like repeat domain (HLR) and a C-terminal domain (CTD) (where a syndecan-binding domain (SBD) is found). Furthermore, the Sec14, PH-like, and HLR domains form part of a so-called leucine-rich domain (LRD). Dimerization sites are found scattered within the N-heat domain (distal N-terminus and TBD), particularly within the C-heat domain. In principle, an MPNST treated according to the present invention can have one or more mutations or genetic changes (particularly one or more inactivating mutations or inactivating genetic changes) in the NF1 gene sequence for any one (or several) of the above domains or segments of the NF1 gene product, neurofibromin. Thus, for example, an MPNST can be an MPNST having one or more inactivating mutations located in the GTPase-activating domain (GAD)-related domain (GRD) of NF1. In some embodiments, the MPNST has one or more inactivating mutations located in the cysteine- and serine-rich domain / GTPase-activating domain (CSRD) of NF1. In some embodiments, the MPNST has one or more inactivating mutations located in the leucine-rich domain (LRD) of NF1.In some embodiments, the MPNST has one or more inactivating mutations located at at least one dimerization interface of NF1.

[0020] The MPNST to be treated according to the present invention may be an MPNST without mutations or genetic changes affecting the NF1 gene, particularly an MPNST having a wild-type NF1 gene. Thus, the MPNST to be treated can be, for example, a sporadic MPNST having a wild-type NF1 gene, or a radiation-induced MPNST having a wild-type NF1 gene.

[0021] Furthermore, the MPNST to be treated according to the present invention has one or more mutations or genetic changes (particularly, one or more inactivating mutations or inactivating genetic changes in the CDKN2A, p53, RB1, PTEN, and / or PRC2 genes) that affect (particularly decrease or suppress) the expression and / or activity of CDKN2A, p53, RB1, PTEN, and / or PRC2 and / or one or more mutations or genetic changes (including, for example, copy number variations) that affect (particularly activate or enhance) the expression and / or activity of EGFR, PDGFRA, and / or c-Met (particularly, one or more activating mutations or activating genetic changes in the EGFR, PDGFRA, and / or c-Met genes), and can be an MPNST (including any one of the specific types of MPNSTs such as type I neurofibromatosis-related MPNST, sporadic MPNST, or radiation-induced MPNST).

[0022] Accordingly, in particular, the MPNST to be treated can be one or more mutations or genetic changes that affect (in particular, reduce or suppress) the expression and / or activity of CDKN2A, in particular one or more inactivating mutations or inactivating genetic changes of CDKN2A, such as an MPNST having one or more inactivating mutations or inactivating genetic changes of CDKN2A (including, for example, any one of the specific types of MPNST such as neurofibromatosis type I-associated MPNST, sporadic MPNST, or radiation-induced MPNST). Accordingly, the MPNST to be treated can be an MPNST associated with inactivation of CDKN2A, such as a neurofibromatosis type I-associated MPNST associated with inactivation of CDKN2A, a sporadic MPNST associated with inactivation of CDKN2A, or a radiation-induced MPNST associated with inactivation of CDKN2A. Such inactivation of CDKN2A (in particular, inactivation of both alleles of CDKN2A) can be due to, for example, one or more point mutations, inversions, deletions, or translocations of CDKN2A (see, for example, Magallon-Lorenz M et al., Hum Genet 2021 140(8):1241-52, doi:10.1007 / s00439-021-02296-x).

[0023] The MPNST to be treated may also be an MPNST having one or more mutations or genetic changes that affect (in particular, reduce or suppress) the expression and / or activity of p53, in particular one or more inactivating mutations or inactivating genetic changes of p53, such as an MPNST having one or more inactivating mutations or inactivating genetic changes of p53 (including, for example, any one of the specific types of MPNST). Accordingly, the MPNST to be treated can be an MPNST associated with inactivation of p53, such as a neurofibromatosis type I-associated MPNST associated with inactivation of p53, a sporadic MPNST associated with inactivation of p53, or a radiation-induced MPNST associated with inactivation of p53.

[0024] The MPNST to be treated may be an MPNST having one or more mutations or genetic changes that affect (particularly decrease or suppress) the expression and / or activity of RB1, particularly one or more inactivating mutations or inactivating genetic changes of RB1 (e.g., including any one of the above-specified types of MPNST). Accordingly, the MPNST to be treated may be an MPNST associated with inactivation of RB1, such as a neurofibromatosis type I-associated MPNST associated with inactivation of RB1, a sporadic MPNST associated with inactivation of RB1, or a radiation-induced MPNST associated with inactivation of RB1.

[0025] The MPNST to be treated may be an MPNST having one or more mutations or genetic changes that affect (particularly decrease or suppress) the expression and / or activity of PTEN, particularly one or more inactivating mutations or inactivating genetic changes of PTEN (e.g., including any one of the above-specified types of MPNST). Accordingly, the MPNST to be treated may be an MPNST associated with inactivation of PTEN, such as a neurofibromatosis type I-associated MPNST associated with inactivation of PTEN, a sporadic MPNST associated with inactivation of PTEN, or a radiation-induced MPNST associated with inactivation of PTEN.

[0026] The MPNST to be treated may be an MPNST having one or more mutations or genetic changes that affect (particularly decrease or suppress) the expression and / or activity of PRC2, particularly one or more inactivating mutations or inactivating genetic changes of PRC2 (e.g., including any one of the above-specified types of MPNST). Accordingly, the MPNST to be treated may be an MPNST associated with inactivation of PRC2, such as a neurofibromatosis type I-associated MPNST associated with inactivation of PRC2, a sporadic MPNST associated with inactivation of PRC2, or a radiation-induced MPNST associated with inactivation of PRC2. Such inactivation of PRC2 may be due to, for example, one or more mutations in one or more PRC2 core component genes, such as EZH2, EED, and / or SUZ12.

[0027] The MPNST to be treated may be an MPNST having one or more mutations or genetic changes that affect (particularly activate or enhance) the expression and / or activity of EGFR, particularly one or more activating mutations or activating genetic changes of EGFR (for example, including any one of the above-specified types of MPNST). Accordingly, the MPNST to be treated may be an MPNST associated with the activation or upregulation of EGFR, such as an MPNST type I neurofibromatosis-associated MPNST associated with the activation / upregulation of EGFR, a sporadic MPNST associated with the activation / upregulation of EGFR, or a radiation-induced MPNST associated with the activation / upregulation of EGFR.

[0028] The MPNST to be treated may be an MPNST having one or more mutations or genetic changes that affect (particularly activate or enhance) the expression and / or activity of PDGFRA, particularly one or more activating mutations or activating genetic changes of PDGFRA (for example, including any one of the above-specified types of MPNST). Accordingly, the MPNST to be treated may be an MPNST associated with the activation or upregulation of PDGFRA, such as an MPNST type I neurofibromatosis-associated MPNST associated with the activation / upregulation of PDGFRA, a sporadic MPNST associated with the activation / upregulation of PDGFRA, or a radiation-induced MPNST associated with the activation / upregulation of PDGFRA.

[0029] The MPNST to be treated may be an MPNST having one or more mutations or genetic changes that affect (particularly activate or enhance) the expression and / or activity of c-Met, particularly one or more activating mutations or activating genetic changes of c-Met (for example, including any one of the above-specified types of MPNST). Accordingly, the MPNST to be treated may be an MPNST associated with the activation or upregulation of c-Met, such as an MPNST type I neurofibromatosis-associated MPNST associated with the activation / upregulation of c-Met, a sporadic MPNST associated with the activation / upregulation of c-Met, or a radiation-induced MPNST associated with the activation / upregulation of c-Met.

[0030] The MPNST to be treated may further be an MPNST associated with chromosome 8 gain (for example, including any one of the above-specified types of MPNST, such as an MPNST type I neurofibromatosis-associated MPNST, a sporadic MPNST, or a radiation-induced MPNST) (see, for example, Dehner C et al., JCI Insight, 2021, 6(6):e146351, doi:10.1172 / jci.insight.146351).

[0031] The MPNST to be treated may also be an MPNST that overexpresses one or more SNAG domain transcription factors, such as Snail1 (Snail), Snail2 (Slug), Snail3 (Smuc), Scratch 1, Scratch 2, Gfi-1, Gfi-1B, Insm1, Insm2, Ovol-1 (Ovo-like 1), Ovol-2, and / or Ovol-3 (for example, including any one of the above-specified types of MPNST).

[0032] Furthermore, the MPNST to be treated may be an MPNST associated with aneuploidy (where one or more chromosomes are present in a copy number of 3 or more and / or an odd copy number) (for example, including any one of the above-specified types of MPNST).

[0033] The MPNST to be treated (including any one of the above-specified types of MPNST) may further be a metastatic MPNST. Thus, the MPNST to be treated may be a primary MPNST that has formed metastases, i.e., has spread to one or more other parts of the subject's body (such as the lungs and / or bones, etc.). In particular, the MPNST may be, for example, a metastatic neurofibromatosis type I-related MPNST, a metastatic sporadic MPNST, or a metastatic radiation-induced MPNST.

[0034] The MPNST to be treated (including any one of the above-specified types of MPNST) may also be a recurrent or refractory MPNST. In particular, the MPNST may be, for example, a recurrent or refractory neurofibromatosis type I-related MPNST, a recurrent or refractory sporadic MPNST, or a recurrent or refractory radiation-induced MPNST.

[0035] The therapeutic effect of the LSD1 inhibitor in the treatment of MPNST can be further confirmed by additional in vitro or in vivo experiments as well as clinical trials in humans, which can be easily set up by those skilled in the art of drug development.

[0036] LSD1 inhibitor As used herein, the term "LSD1 inhibitor" means a compound / substance that reduces, decreases, blocks or inhibits the gene expression, activity or function of LSD1. Compounds that act as inhibitors of LSD1 are known in the art. Any molecule that acts as an LSD1 inhibitor can in principle be used in the context of the present invention. Preferably, the LSD1 inhibitor is a small molecule. Furthermore, the LSD1 inhibitor can be an irreversible LSD1 inhibitor or a reversible LSD1 inhibitor. As also shown in Example 1 below, both irreversible and reversible LSD1 inhibitors can be used for the treatment of MPNST according to the present invention. Typical irreversible LSD1 inhibitors are cyclopropylamine-based compounds such as iadademstat and bomedemstat, and among the LSD1 inhibitors, these are used in Example 1. A representative example of a reversible LSD1 inhibitor is the compound prurodemstat, which is also used in Example 1. Preferably, the LSD1 inhibitor is a selective LSD1 inhibitor, and as used herein, the term "selective LSD1 inhibitor" means an LSD1 inhibitor that exhibits at least 10-fold (preferably at least 100-fold) selectivity for LSD1 over other FAD-dependent monoamine oxidases, particularly MAO-A and MAO-B (this can be evaluated, for example, by determining the IC 50 values).

[0037] An exemplary list of small molecule LSD1 inhibitors is provided in the table below.

[0038]

Table A

[0039] The LSD1 inhibitor used according to the present invention can thus be, for example, any one of the specific compounds listed in the table above, or a pharmaceutically acceptable salt of any one of these compounds.

[0040] In some embodiments, the LSD1 inhibitor is, for example, WO2010 / 043721, WO2010 / 084160, WO2010 / 143582, WO2011 / 035941, WO2011 / 042217, WO2011 / 131576, WO2011 / 131697, WO2012 / 013727, WO2012 / 013728, WO2012 / 045883, WO2012 / 135113, WO2013 / 022047, EP2743256A1, WO2013 / 025805, WO2013 / 057320, WO2013 / 057322, WO2014 / 058071, EP2907802A1, WO2014 / 084298, EP2927212A1, WO2014 / 086790, WO2014 / 164867, WO2014 / 194280, WO2014 / 205213, WO2015 / 021128, WO2015 / 031564, WO2015 / 089192, WO2015 / 120281, WO2015 / 123408, WO2015 / 123424, WO2015 / 123437, WO2015 / 123465, WO2015 / 134973, WO2015 / 168466, WO2015 / 181380, WO2015 / 200843, WO2016 / 003917, WO2016 / 004105, WO2016 / 007722, WO2016 / 007727, WO2016 / 007731, WO2016 / 007736, WO2016 / 034946, WO2016 / 037005, WO2016 / 123387, WO2016 / 130952, WO2016 / 161282, WO2016 / 172496, WO2016 / 177656, WO2017 / 004519, WO2017 / 027678, WO2017 / 079476, WO2017 / 079670, WO2017 / 090756, EP3381896A1, WO2017 / 109061, WO2017 / 116558, WO2017 / 149463, WO2017 / 157322, EP3431471A1, WO2017 / 184934, WO2017 / 195216, WO2017 / 198780, WO2017 / 215464, EP3486244A1, WO2018 / 081342, WO2018 / 081343, WO2018 / 137644, EP3575285A1, WO2018 / 213211,Any one of the compounds disclosed in WO2018 / 216800, EP3632897A1, WO2018 / 226053, WO2018 / 234978, WO2019 / 009412, WO2019 / 034774, WO2019 / 054766, WO2019 / 217972, WO2019 / 222069, WO2020 / 015745, EP3825309A1, WO2020 / 047198, WO2020 / 052647, WO2020 / 052649, EP3851440A1, WO2020 / 138398, WO2020 / 159285, EP3907225A1, WO2021 / 058024, WO2021 / 095835, WO2021 / 175079, WO2022 / 072811, WO2022 / 171044, WO2022 / 188709, WO2022 / 240886, WO2022 / 267495, WO2023 / 069884, WO2023 / 284651, US2017-0283397, US2022-0064126, CN103054869, CN103319466, CN104119280, CN105541806, CN105924362, CN105985265, CN106045862, CN106045881, CN106432248, CN106478639, CN106831489, CN106928235, CN107033148 CN107174584, CN107176927, CN107459476, CN107474011, CN107501169, CN107936022, CN108530302, CN109265462, CN109293664, CN109535019, CN110204551, CN110478352, CN111072610, CN111454252, CN112110936, CN112409310, CN112920130, CN113087712, CN113105479, CN113264903, CN113582906, CN113599380, CN114502561, CN114805205, CN114805261, KR20190040763, or KR20190040783 is an LSD1 inhibitor known in the art. Each of the above documents is incorporated herein by reference in its entirety (in particular,(including the compounds described in the Examples section of each of these documents). Therefore, the LSD1 inhibitor may be, for example, a compound disclosed in any one of the above-mentioned documents (including, for example, the Examples section of any one of these documents), and the compound can be used in the form of a non-salt form or a pharmaceutically acceptable salt.,

[0041] In some embodiments, the LSD1 inhibitor is selected from the group consisting of iadademstat, prolademstat, bomedemstat, seclidemstat, 1-((4-(methoxymethyl)-4-(((1R,2S)-2-phenylcyclopropylamino)methyl)piperidin-1-yl)methyl)cyclobutanecarboxylic acid, 3-(cyanomethyl)-3-(4-{[(1R,2S)-2-phenylcyclopropyl]amino}piperidin-1-yl)azetidine-1-sulfonamide, vafidemstat, 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile, and pharmaceutically acceptable salts thereof (i.e., pharmaceutically acceptable salts of any one of the aforementioned compounds).

[0042] Iadademstat is a selective and irreversible LSD1 inhibitor. Iadademstat has the formula:

[0043]

Chemical formula

[0044] The INN (International Nonproprietary Name) of the compound with [CAS Registry Number 1431304-21-0], also known as ORY-1001 or (trans)-N1-((1R,2S)-2-phenylcyclopropyl)cyclohexane-1,4-diamine. Iadademstat is described, for example, in Example 5 of WO2013 / 057322. Pharmaceutically acceptable salts of iadademstat, including the hydrochloride salt (especially iadademstat dihydrochloride), are also described in WO2013 / 057322.

[0045] Purademstat has the formula:

[0046]

Chemical formula

[0047] A reversible LSD1 inhibitor with [CAS Registry Number 1821307-10-1], also known as CC-90011, having the chemical name 4-[2-(4-aminopiperidin-1-yl)-5-(3-fluoro-4-methoxyphenyl)-1-methyl-6-oxo-1,6-dihydropyrimidin-4-yl]-2-fluorobenzonitrile. Purademstat is described, for example, in WO2015 / 168466 and WO2017 / 79670. Its pharmaceutically acceptable salts, including the besylate salt, are also described therein.

[0048] Bomedemstat has the formula:

[0049]

Chemical formula

[0050] An irreversible LSD1 inhibitor with [CAS Registry Number 1990504-34-1], also known as IMG-7289, having the chemical name N-[(2S)-5-{[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino}-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide. Bomedemstat is described, for example, in WO2016 / 130952 and WO2018 / 35259. Its pharmaceutically acceptable salts, including the bis tosylate salt, are also described therein.

[0051] Secdemstat has the formula:

[0052]

Chem.

[0053] An LSD1 inhibitor with [CAS Registry Number 1423715-37-0], also known as SP-2577, having the chemical name (E)-N’-(1-(5-chloro-2-hydroxyphenyl)ethylidene)-3-((4-methylpiperazin-1-yl)sulfonyl)benzohydrazide. Secdemstat is described, for example, in WO2013 / 025805 and WO2014 / 205213.

[0054] 1-((4-(methoxymethyl)-4-(((1R,2S)-2-phenylcyclopropylamino)methyl)piperidin-1-yl)methyl)cyclobutanecarboxylic acid is an irreversible LSD1 inhibitor and is described, for example, in WO2015 / 123465 and WO2017 / 27678. Its pharmaceutically acceptable salts, including the paratoluenesulfonate salt, are also described therein. The structure of this compound can be depicted as follows.

[0055]

Chem.

[0056] 3-(Cyanomethyl)-3-(4-{[(1R,2S)-2-phenylcyclopropyl]amino}piperidin-1-yl)azetidine-1-sulfonamide is an irreversible LSD1 inhibitor and is described, for example, in WO2020 / 047198. Its pharmaceutically acceptable salts are also described therein. The structure of this compound can be depicted as follows.

[0057] [Chemical formula]

[0058] Bafidemstat is an irreversible LSD1 inhibitor of the formula:

[0059] [Chemical formula]

[0060] and is also known as ORY-2001, 5-((((1R,2S)-2-(4-(benzyloxy)phenyl)cyclopropyl)amino)methyl)-1,3,4-oxadiazol-2-amine, or (-) 5-((((trans)-2-(4-(benzyloxy)phenyl)cyclopropyl)amino)methyl)-1,3,4-oxadiazol-2-amine. Bafidemstat is described, for example, in Example 35 of WO2012 / 13728.

[0061] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile is an LSD1 inhibitor described, for example, in WO2017 / 090756 (or EP3381896A1; see Example 37), WO2021 / 095835, WO2022 / 240886, and WO2023 / 054547. Among them are pharmaceutically acceptable salts of this compound, including benzoate (or benzoyl salt), sorbate, succinate, L-tartrate, hydrochloride, hemifumarate salt, monofumarate salt, hemioxalate salt, monooxalate salt, mesylate salt, esylate salt, or maleate salt. A specific solid form of this compound is described in WO2022 / 240886. This compound is also referred to herein as "TAS1440". The structure of this compound can be depicted as follows.

[0062] [Chemical formula]

[0063] Further examples of LSD1 inhibitors include SYHA1807 or its pharmaceutically acceptable salts, or JBI-802 or its pharmaceutically acceptable salts. In some embodiments, the LSD1 inhibitor is selected from the group consisting of iadademstat, prulodemstat, bomedemstat, seclidemstat, 1-((4-(methoxymethyl)-4-(((1R,2S)-2-phenylcyclopropylamino)methyl)piperidin-1-yl)methyl)cyclobutanecarboxylic acid, 3-(cyanomethyl)-3-(4-{[(1R,2S)-2-phenylcyclopropyl]amino}piperidin-1-yl)azetidine-1-sulfonamide, 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile, and their pharmaceutically acceptable salts.

[0064] In particular, the LSD1 inhibitor can be selected from the group consisting of iadademstat, prulodemstat, bomedemstat, seclidemstat, 1-((4-(methoxymethyl)-4-(((1R,2S)-2-phenylcyclopropylamino)methyl)piperidin-1-yl)methyl)cyclobutanecarboxylic acid, 3-(cyanomethyl)-3-(4-{[(1R,2S)-2-phenylcyclopropyl]amino}piperidin-1-yl)azetidine-1-sulfonamide, and pharmaceutically acceptable salts thereof.

[0065] In a preferred embodiment, the LSD1 inhibitor is selected from the group consisting of iadademstat, prulodemstat, bomedemstat, and pharmaceutically acceptable salts thereof. In some embodiments, the LSD1 inhibitor is prulodemstat or a pharmaceutically acceptable salt thereof (e.g., prulodemstat besilate). In some embodiments, the LSD1 inhibitor is bomedemstat, or a pharmaceutically acceptable salt thereof (e.g., bomedemstat bis tosylate).

[0066] A particularly preferred LSD1 inhibitor is iadademstat or a pharmaceutically acceptable salt thereof. In some embodiments, iadademstat is used as the dihydrochloride salt (i.e., iadademstat dihydrochloride).

[0067] Unless otherwise specified, throughout this specification and the claims, any reference to an LSD1 inhibitor (e.g., iadademstat) includes not only the LSD1 inhibitor in non-salt form but also any pharmaceutically acceptable salt thereof. When the LSD1 inhibitor is iadademstat, it is preferably used in the form of a pharmaceutically acceptable salt, preferably the hydrochloride salt, more preferably the dihydrochloride salt.

[0068] Pharmaceutical preparation The LSD1 inhibitors used in accordance with the present invention, as well as any pharmaceutical composition comprising an LSD1 inhibitor used in accordance with the present invention, can be administered by any route appropriate to the condition being treated. Exemplary routes include oral, parenteral (including subcutaneous, intramuscular, intravenous, intraarterial, inhalation, intradermal, intrathecal, epidural, and infusion techniques), transdermal, rectal, nasal, topical (including buccal and sublingual), vaginal, intraperitoneal, intrapulmonary, and intranasal. Preferably, the LSD1 inhibitor (or its corresponding pharmaceutical composition) is administered orally.

[0069] The LSD1 inhibitors used in accordance with the present invention can be administered in any convenient pharmaceutical composition or formulation, such as, for example, tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions / formulations can contain conventional ingredients in pharmaceutical manufacture, such as diluents, carriers, pH adjusters, preservatives, solubilizers, stabilizers, wetting agents, emulsifying agents, sweeteners, colorants, flavoring agents, salts for altering osmotic pressure, buffers, masking agents, antioxidants, and / or additional active drugs. They can further contain other therapeutically effective or therapeutically valuable substances.

[0070] Typical formulations are manufactured by mixing an LSD1 inhibitor with one or more pharmaceutically acceptable excipients. Suitable excipients are well known to those skilled in the art and are described in detail, for example, in “Ansel’s Pharmaceutical Dosage Forms and Drug Delivery Systems” (2004) Lippincott, Williams & Wilkins, Philadelphia; “Remington: The Science and Practice of Pharmacy” (2000) Lippincott, Williams & Wilkins, Philadelphia; or “Handbook of Pharmaceutical Excipients” (2005) Pharmaceutical Press, Chicago. The formulations may also include one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, flow promoters, processing aids, colorants, sweeteners, fragrances, flavoring agents, diluents, and / or other known additives for improving the appearance of the active drug or for assisting in the manufacture of a pharmaceutical (i.e., a medicine).

[0071] For oral delivery, the LSD1 inhibitor can be formulated in a preparation containing a pharmaceutically acceptable carrier, such as a binder (e.g., gelatin, cellulose, or tragacanth gum), an excipient (e.g., starch or lactose), a lubricant (e.g., magnesium stearate or silicon dioxide), a disintegrant (e.g., alginate, Primogel, or corn starch), and a sweetener or flavoring agent (e.g., glucose, sucrose, saccharin, methyl salicylate, or peppermint). The preparation can be administered orally, for example, in the form of a sealed gelatin capsule or a compressed tablet. Capsules and tablets can be manufactured by any conventional technique. Capsules and tablets can also be coated with various coatings known in the art to modify the flavor, taste, color, and shape of the capsules and tablets. Further, a liquid carrier such as a fatty oil can be included in the capsule.

[0072] Suitable oral formulations may be in the form of suspensions, syrups, chewing gums, wafers, elixirs, etc. If desired, conventional agents for modifying the flavor, taste, color, and shape of the special form may also be included. Further, when it is convenient to administer to a subject who cannot swallow by means of an enteral nutrition tube, the active compound can also be dissolved in an acceptable lipophilic vegetable oil vehicle, such as olive oil, corn oil, or safflower oil.

[0073] The LSD1 inhibitor can also be administered parenterally in the form of a solution or suspension, or in a lyophilized form that can be converted to a solution or suspension before use. In such formulations, diluents or pharmaceutically acceptable carriers, such as sterile water and physiological saline buffers, can be used. All other conventional solvents, pH buffers, stabilizers, antibacterial agents, surfactants, and antioxidants can also be included. For example, as active ingredients, sodium chloride, acetic acids, citrate or phosphate buffers, glycerin, dextrose, fixed oils, methylparaben, polyethylene glycol, propylene glycol, sodium bisulfate, benzyl alcohol, ascorbic acid, etc. can be mentioned. The parenteral formulation can be stored in any conventional container, such as vials and ampoules.

[0074] Subcutaneous implantation for sustained release of LSD1 inhibitors may also be a suitable route of administration. This would require a surgical procedure for implanting an LSD1 inhibitor in any suitable formulation into a subcutaneous cavity, such as beneath the lower anterior abdominal wall. See, for example, Wilson et al., (1984) J. Clin. Psych. 45:242-247. Hydrogels can be used as carriers for the sustained release of LSD1 inhibitors. Hydrogels are generally known in the art. These are typically produced by crosslinking high molecular weight biocompatible polymers into a network, which swells with water to form a gel-like substance. Preferably, the hydrogel is biodegradable or bioabsorbable. For the purposes of the present invention, hydrogels made of polyethylene glycol, collagen, or poly(glycol-co-L-lactic acid) may be useful. See, for example, Phillips et al., (1984) J. Pharmaceut. Sci., 73:1718-1720.

[0075] Pharmaceutical compositions, such as oral and parenteral compositions, can be formulated into unit dosage forms for ease of administration and uniform dosage. As used herein, "unit dosage form" refers to a physically discrete unit suitable as a unit dose for administration to a subject, each unit containing a predetermined amount of an active ingredient calculated to produce the desired therapeutic effect, together with one or more suitable pharmaceutical carriers.

[0076] Suitable oral dosage forms of iadademstat are disclosed, for example, in WO2019 / 211491. In particular, iadademstat can be provided in the form of solid oral dosage forms, such as tablets or capsules. Alternatively, iadademstat may be provided in the form of an oral liquid composition, particularly an oral solution, such as an aqueous oral solution (the corresponding oral solution containing an aqueous oral solution can be prepared, for example, from a powder for reconstitution). As described above, iadademstat is preferably used in the form of iadademstat dihydrochloride.

[0077] In the case of treating MPNST, the LSD1 inhibitor (or the corresponding pharmaceutical composition) can be administered in any suitable manner as determined by an expert in the medical field. The appropriate dosage, administration period, and dosing frequency can vary within a wide range and are determined, inter alia, by factors such as the condition of the subject, the specific type and severity of the disease, the particular form of the active ingredient, and the method of administration. Generally, with an appropriate dosage and administration schedule, the LSD1 inhibitor is provided in an amount sufficient to provide a therapeutic benefit, such as an improved clinical outcome, such as more frequent complete or partial remission, or a longer disease-free period and / or overall survival, or a reduction in the severity of symptoms, or any other objectively confirmable improvement as noticed by the clinician. The therapeutically effective amount can generally be evaluated or estimated using experimental models such as dose-response curves derived from in vitro or animal model test systems, or from human clinical trials.

[0078] As is well known to those skilled in the art, the appropriate dosage and administration schedule of the LSD1 inhibitor depend on the specific LSD1 inhibitor used, its LSD1 inhibitory ability, its pharmacokinetic profile, and other factors.

[0079] Iadademstat is a highly potent active pharmaceutical ingredient (HPAPI). Thus, the expected daily dose is very low, for example less than 1 mg per day. Accordingly, the drug load in pharmaceutical formulations (including, for example, solid oral forms) is also typically very low (for example, less than 1 mg of API per 100 mg of solid oral form). Generally, for oral administration to adult human subjects (i.e., human subjects 18 years of age or older), the daily dose is about 50 μg to about 300 μg, preferably about 75 μg to about 300 μg (for example, about 75 μg, about 100 μg, about 125 μg, about 150 μg, about 175 μg, about 200 μg, about 225 μg, about 250 μg, about 275 μg, or about 300 μg, or any range between any two of the foregoing daily doses), of iadademstat as described herein, although these limits can be adjusted if necessary. For example, the foregoing doses can be reduced in the case of oral administration to pediatric subjects, particularly human subjects less than 18 years of age (for example, 0 to 2 years, 2 to 12 years, or less than 12 to 18 years). As used herein, the term “μg” (or “ug”) refers to microgram.

[0080] In some embodiments, the LSD1 inhibitor is iadademstat (or a pharmaceutically acceptable salt thereof, such as iadademstat dihydrochloride), and is administered on a five days on / two days off (5 / 2) schedule.

[0081] In some embodiments, the LSD1 inhibitor is iadademstat (or a pharmaceutically acceptable salt thereof, such as iadademstat dihydrochloride), and is orally administered to adult human subjects at a daily dose of about 50 μg to about 300 μg, preferably about 75 μg to about 300 μg (e.g., about 100 μg to about 300 μg), on a 5-day dosing / 2-day rest (5 / 2) schedule. The doses reflected herein for iadademstat relate to the corresponding amounts of the iadademstat free base. In some embodiments, iadademstat is orally administered at a daily dose of about 75 μg on a 5-day dosing / 2-day rest (5 / 2) schedule. In some embodiments, iadademstat is orally administered at a daily dose of about 100 μg on a 5-day dosing / 2-day rest (5 / 2) schedule. In some embodiments, iadademstat is orally administered at a daily dose of about 150 μg on a 5-day dosing / 2-day rest (5 / 2) schedule. In some embodiments, iadademstat is orally administered at a daily dose of about 200 μg on a 5-day dosing / 2-day rest (5 / 2) schedule. In some embodiments, iadademstat is orally administered at a daily dose of about 250 μg on a 5-day dosing / 2-day rest (5 / 2) schedule. In some embodiments, iadademstat is orally administered at a daily dose of about 300 μg on a 5-day dosing / 2-day rest (5 / 2) schedule. As described above, these doses can be reduced for pediatric use.

[0082] Combination therapy The LSD1 inhibitor used in accordance with the present invention can be administered in monotherapy (single-agent therapy) (e.g., without co-administering any additional therapeutic agent or without co-administering any additional anti-cancer agent). Accordingly, the present invention relates to an LSD1 inhibitor (or a pharmaceutical composition comprising an LSD1 inhibitor and optionally one or more pharmaceutically acceptable excipients) for use in monotherapy of MPNST. The present invention similarly relates to the corresponding method and use for monotherapy of MPNST.

[0083] However, the LSD1 inhibitor can also be administered in combination with one or more additional therapeutic agents, particularly one or more additional anti-cancer agents. When the LSD1 inhibitor is used in combination with an additional anti-cancer agent, the dosage of each compound may be different from that when the corresponding compound is used alone. In particular, one or both of the compounds can be used at a low dosage.

[0084] The combination of an LSD1 inhibitor and one or more additional therapeutic agents (such as one or more additional anti-cancer agents) can include co-administering / co-dosing the LSD1 inhibitor and the additional therapeutic agent either in a single pharmaceutical formulation or in separate pharmaceutical formulations, or administering / dosing the LSD1 inhibitor and the additional therapeutic agent sequentially / separately. If the administration is sequential, either the LSD1 inhibitor or one or more of the additional therapeutic agents can be administered first. If the administration is simultaneous, the one or more additional therapeutic agents can be included in the same pharmaceutical formulation as the LSD1 inhibitor or can be administered in two or more distinct / separate pharmaceutical formulations. It will also be understood that administering the LSD1 inhibitor and the additional therapeutic agent in separate pharmaceutical formulations can be advantageous, for example, when each agent is administered using a different route and / or a different dosing schedule / regimen.

[0085] Furthermore, the LSD1 inhibitor can also be administered in combination with physical therapy, particularly radiation therapy. The present invention also relates to the combined use of an LSD1 inhibitor and one or more additional therapeutic agents (particularly one or more additional anti-cancer agents) and physical therapy (particularly radiation therapy). Physical therapy (or radiation therapy) can be initiated before, after, or simultaneously with the administration of the LSD1 inhibitor (e.g., about 1 to 72 hours before or after the administration of the LSD1 inhibitor).

[0086] Accordingly, the present invention relates to an LSD1 inhibitor for use in the treatment of MPNST in combination with one or more additional therapeutic agents (particularly one or more additional anti-cancer agents) and / or in combination with radiotherapy. The present invention also relates to a pharmaceutical composition comprising an LSD1 inhibitor and optionally one or more pharmaceutically acceptable excipients for use in the treatment of MPNST in combination with one or more additional therapeutic agents (particularly one or more additional anti-cancer agents) and / or in combination with radiotherapy. The present invention further relates to an LSD1 inhibitor (or a pharmaceutical composition comprising an LSD1 inhibitor and optionally one or more pharmaceutically acceptable excipients) for use in the treatment of MPNST, wherein the LSD1 inhibitor (or the pharmaceutical composition comprising the LSD1 inhibitor) is administered in combination with one or more additional therapeutic agents (particularly one or more additional anti-cancer agents) and / or in combination with radiotherapy. The present invention further relates to an LSD1 inhibitor (or a pharmaceutical composition comprising an LSD1 inhibitor and optionally one or more pharmaceutically acceptable excipients) for use in the treatment of MPNST, wherein the LSD1 inhibitor (or the pharmaceutical composition comprising the LSD1 inhibitor) is used in combination with one or more additional therapeutic agents (particularly one or more additional anti-cancer agents) and / or in combination with radiotherapy. The present invention further relates to (i) an anti-cancer agent for use in the treatment of MPNST in combination with an LSD1 inhibitor; (ii) an anti-cancer agent for use in the treatment of MPNST (the anti-cancer agent being co-administered with an LSD1 inhibitor); or (iii) an anti-cancer agent for use in the treatment of MPNST (the anti-cancer agent being used in combination with an LSD1 inhibitor).

[0087] The present invention also provides a method of treating MPNST in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of an LSD1 inhibitor (or a therapeutically effective amount of a pharmaceutical composition comprising an LSD1 inhibitor and optionally one or more pharmaceutically acceptable excipients) in combination with a therapeutically effective amount of one or more additional therapeutic agents (particularly one or more additional anti-cancer agents) and / or in combination with radiotherapy.

[0088] Furthermore, the present invention also relates to the use of an LSD1 inhibitor for the treatment of MPNST in combination with one or more further therapeutic agents (in particular one or more further anti-cancer agents) and / or in combination with radiotherapy. The present invention also relates to the use of an anti-cancer agent for the treatment of MPNST in combination with an LSD1 inhibitor.

[0089] The present invention further also relates to the use of an LSD1 inhibitor for manufacturing a medicament (or pharmaceutical composition) for treating MPNST in combination with one or more further therapeutic agents (in particular one or more further anti-cancer agents) and / or in combination with radiotherapy. The present invention also relates to the use of an anti-cancer agent for manufacturing a medicament for treating MPNST in combination with an LSD1 inhibitor. The present invention similarly also relates to the use of an LSD1 inhibitor and one or more further anti-cancer agents for manufacturing a medicament for treating MPNST, wherein the medicament in this case comprises the LSD1 inhibitor and the further anti-cancer agent(s) in the same pharmaceutical formulation or in separate pharmaceutical formulations. The present invention also relates to the use of an LSD1 inhibitor for manufacturing a medicament for treating MPNST, wherein the medicament in this case is manufactured for use in combination (or in conjunction) with one or more further therapeutic agents (in particular one or more further anti-cancer agents) and / or radiotherapy. The present invention also relates to the use of an anti-cancer agent for manufacturing a medicament for treating MPNST, wherein the medicament in this case is manufactured for use in combination (or in conjunction) with an LSD1 inhibitor.

[0090] The present invention further also provides a combined product comprising an LSD1 inhibitor and one or more further therapeutic agents (in particular one or more further anti-cancer agents) in the same pharmaceutical formulation or in separate pharmaceutical formulations for use in the treatment of MPNST. Thus, the LSD1 inhibitor and the further therapeutic agent (in particular the further anti-cancer agent(s)) may be present in a single pharmaceutical formulation (i.e., in the same pharmaceutical formulation) or may be provided in separately distinct (separate) pharmaceutical formulations.

[0091] The present invention also provides a pharmaceutical composition comprising an LSD1 inhibitor in combination with one or more additional therapeutic agents (particularly one or more additional anti-cancer agents) for use in the treatment of MPNST, and one or more pharmaceutically acceptable excipients.

[0092] The present invention further provides an article of manufacture (or kit) comprising an LSD1 inhibitor and one or more additional therapeutic agents (particularly one or more additional anti-cancer agents) in the same pharmaceutical formulation or in separate pharmaceutical formulations for use in the treatment of MPNST.

[0093] The present invention further provides a method of treating MPNST in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the above combination product, pharmaceutical composition or article of manufacture. In particular, the present invention provides a method of treating MPNST in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a combination product comprising an LSD1 inhibitor and one or more additional therapeutic agents (particularly one or more additional anti-cancer agents) in the same pharmaceutical formulation or in separate pharmaceutical formulations. The present invention further provides a method of treating MPNST in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of an LSD1 inhibitor and a therapeutically effective amount of one or more additional therapeutic agents (particularly one or more additional anti-cancer agents).

[0094] The present invention further provides the use of a combination comprising an LSD1 inhibitor and one or more additional therapeutic agents (particularly one or more additional anti-cancer agents) for the manufacture of a medicament (or pharmaceutical composition) for treating MPNST. The present invention also provides the use of a combination comprising an LSD1 inhibitor and one or more additional therapeutic agents (particularly one or more additional anti-cancer agents) for treating MPNST.

[0095] The anticancer agent(s) (in particular, the "anticancer agent" or "one or more additional anticancer agents" mentioned in any of the above paragraphs) are, for example, MEK inhibitors (in particular, inhibitors of MEK1 and / or MEK2; for example, selumetinib), Pi3K inhibitors (for example, copanlisib), mTOR inhibitors (for example, temsirolimus), ERK inhibitors (in particular, inhibitors of ERK1 and / or ERK2; for example, ulixertinib), kRAS inhibitors (for example, sotorasib), EGFR inhibitors (for example, lapatinib), cKIT inhibitors (for example, imatinib), proteasome inhibitors (for example, bortezomib), DNA intercalators (for example, doxorubicin), RAF inhibitors (in particular, BRAF inhibitors; for example, sorafenib), VEGFR inhibitors (for example, cabozantinib), ALK inhibitors (for example, crizotinib), glutaminase inhibitors (for example, telaglenastat), JAK inhibitors (or Janus kinase inhibitors; for example, tofacitinib), PLK1 inhibitors (for example, volasertib), Bcl2 inhibitors (for example, venetoclax), HDAC inhibitors (for example, vorinostat), HSP90 inhibitors (for example, onalespib), Wnt / β-catenin pathway inhibitors (for example, OMP-18R5), aurora kinase inhibitors (for example, alisertib), MDM2 inhibitors (for example, alrizomadlin), CDK4 / 6 inhibitors (for example, abemaciclib), YAP / TAZ pathway inhibitors (for example, pazopanib), SOS inhibitors (for example, BI 1701963), Grb2 inhibitors (for example, BP1001), BET inhibitors (in particular, including BRD4 inhibitors;For example, it can be selected from GSK1210151A), an AKT inhibitor (for example, ipatasertib), a MNK inhibitor (for example, ETC-206), an NTRK inhibitor (for example, entrectinib), a SPH2 inhibitor (for example, JAB-3068), and a PP2A inhibitor (for example, LB100).;

[0096] The MEK inhibitor can be, for example, selumetinib, trametinib, cobimetinib, binimetinib, mirdametinib, pimasertib, refametinib, zapnometinib, avutometinib, HL-085, FCN-159, TAK-733, or a pharmaceutically acceptable salt of any one of these agents. The Pi3K inhibitor can be, for example, copanlisib, alpelisib, idelalisib, duvelisib, umbralisib, buparlisib, zandelisib, linperlisib, parsaclisib, leniolisib, paxalisib, inavolisib, serabelisib, pictilisib, taselisib, tenalisib, eganelisib, GSK2636771, MEN1611, AMG-319, or a pharmaceutically acceptable salt of any one of these agents. The mTOR inhibitor can be, for example, temsirolimus, everolimus, sirolimus, or a pharmaceutically acceptable salt of any one of these agents. The ERK inhibitor can be, for example, ulixertinib or a pharmaceutically acceptable salt thereof. The kRAS inhibitor can be, for example, sotorasib, adagrasib, or a pharmaceutically acceptable salt of any one of these agents. The EGFR inhibitor can be, for example, lapatinib, gefitinib, erlotinib, osimertinib, afatinib, or a pharmaceutically acceptable salt of any one of these agents.The cKIT inhibitor can be, for example, imatinib, sorafenib, lapatinib, sunitinib, or a pharmaceutically acceptable salt of any one of these agents. The proteasome inhibitor can be, for example, bortezomib, carfilzomib, ixazomib, or a pharmaceutically acceptable salt of any one of these agents. The DNA intercalator can be, for example, doxorubicin, daunorubicin, epirubicin, idarubicin, or a pharmaceutically acceptable salt of any one of these agents. The RAF inhibitor can be, for example, sorafenib, encorafenib, dabrafenib, vemurafenib, or a pharmaceutically acceptable salt of any one of these agents. The VEGFR inhibitor can be, for example, cabozantinib, axatinib, lenvatinib, nintedanib, pazopanib, regorafenib, sorafenib, sunitinib, vandetanib, or a pharmaceutically acceptable salt of any one of these agents. The ALK inhibitor can be, for example, crizotinib, alectinib, ceritinib, or a pharmaceutically acceptable salt of any one of these agents. The glutaminase inhibitor can be, for example, telaglenastat or its pharmaceutically acceptable salt. The JAK inhibitor can be, for example, tofacitinib, ruxolitinib, upadacitinib, abrocitinib, or a pharmaceutically acceptable salt of any one of these agents. The PLK1 inhibitor can be, for example, volasertib, onvansertib, rigosertib, BI 2536, or a pharmaceutically acceptable salt of any one of these agents.The Bcl2 inhibitor can be, for example, venetoclax, navitoclax, obatoclax, or a pharmaceutically acceptable salt of any one of these agents. The HDAC inhibitor can be, for example, vorinostat, belinostat, panobinostat, romidepsin, practinostat, rocilinostat, quisinostat, abexinostat, resminostat, givinostat, entinostat, mocetinostat, or a pharmaceutically acceptable salt of any one of these agents. The HSP90 inhibitor can be, for example, onalespib, luminespib, ganetespib, geldanamycin, IPI-504, tanespimycin, alvespimycin, or a pharmaceutically acceptable salt of any one of these agents. The Wnt / β-catenin pathway inhibitor can be, for example, OMP-18R5, OMP-54F28, OTSA 101, SAH-BCL9, XAV939, IWR1, JW74, J01-017a, PKF115-584, PKF118-310, NCB-0846, LGK974, CWP232291, PRI-724, sulindac, vismodegib, glasdegib, or a pharmaceutically acceptable salt of any one of these agents. The aurora kinase inhibitor can be, for example, alisertib, tozasertib, barasertib, danusertib, or a pharmaceutically acceptable salt of any one of these agents.The MDM2 inhibitor can be, for example, alrizomadoline, idasanutlin, RO5045337, RO5503781, AMG232, CGM097, SAR405838, MK-8242, ALRN-6924, or a pharmaceutically acceptable salt of any one of these agents. The CDK4 / 6 inhibitor can be, for example, abemaciclib, ribociclib, palbociclib, or a pharmaceutically acceptable salt of any one of these agents. The YAP / TAZ pathway inhibitor can be, for example, K-975, TED-347, pazopanib, or a pharmaceutically acceptable salt of any one of these agents. The SOS inhibitor can be, for example, BI 1701963, BI 3406, BAY-293, or a pharmaceutically acceptable salt of any one of these agents. The Grb2 inhibitor can be, for example, BP1001, CGP78850, CGP85793, or a pharmaceutically acceptable salt of any one of these agents. The BET inhibitor can be, for example, ABBV-075, ABBV-744, AZD5153, BAY1238097, CPI-203, CPI-0610, GSK1210151A (or I-BET 151), GSK1324726A (I-BET 726), GSK525762 (or I-BET 762), JQ1, LY294002, MS 436, MS 645, MT-1, olinone, OTX-015, RVX-208, TEN-010, or a pharmaceutically acceptable salt of any one of these agents. The AKT inhibitor can be, for example, ipatasertib, uprosertib, afuresertib, MK-2206, triciribine, lactoquinomycin, AZD5363, miransertib, capibasertib, or a pharmaceutically acceptable salt of any one of these agents. The MNK inhibitor can be, for example, ETC-206, SEL-201, BAY1143269, tomivosertib, CGP57380, or a pharmaceutically acceptable salt of any one of these agents.The NTRK inhibitor can be, for example, entrectinib, larotrectinib, or a pharmaceutically acceptable salt of any one of these agents. The SHP2 inhibitor can be, for example, JAB-3068, TNO155, SHP099, RMC-4550, IACS-13909, or a pharmaceutically acceptable salt of any one of these agents. The PP2A inhibitor can be, for example, LB100, cantharidin, cantharidic acid, cytostatin, fostriecin, or a pharmaceutically acceptable salt of any one of these agents. In principle, any of the above anticancer agents can be used in the non-salt form or in the form of a pharmaceutically acceptable salt. The present invention relates specifically and individually to each LSD1 inhibitor described herein in combination with each of the above anticancer agents.

[0097] As described above, one or more additional therapeutic agents used in combination with an LSD1 inhibitor according to the present invention can be (or can include) one or more additional anticancer agents. Alternatively, or additionally, one or more additional therapeutic agents may include an antiemetic. Accordingly, the present invention also relates to an LSD1 inhibitor for use in the treatment of MPNST in combination with one or more additional anticancer agents and also in combination with an antiemetic (and optionally also in combination with radiotherapy); the present invention similarly relates to corresponding methods and uses (including all of the above methods and uses) comprising the combined administration of an LSD1 inhibitor, one or more additional anticancer agents, and an antiemetic. The antiemetic can be, for example, 5-HT 3An antagonist (or “setron”), for example, palonosetron (optionally in combination with netupitant), ramosetron, alosetron, ondansetron, tropisetron, granisetron, dolasetron, azasetron, bemesetron, cilansetron, lerisetron, ricasetron, or zatosetron; olanzapine; a corticosteroid, for example, methylprednisolone or dexamethasone; or prochlorperazine may be used.

[0098] Manufactured article The pharmaceutical composition (or formulation) of the present invention can be placed in a container, pack, or dispenser, together with instructions for administration.

[0099] Accordingly, in a further aspect, the present invention provides an article of manufacture containing an LSD1 inhibitor or a pharmaceutical composition containing an LSD1 inhibitor for treating MPNST as described herein.

[0100] In some aspects, the article of manufacture includes a container and a pharmaceutical composition for use in accordance with the present invention as described herein. In some aspects, the present invention provides an article of manufacture (or kit) including a container and a combination product (as described above) for use in the treatment of MPNST. The present invention also provides an article of manufacture (or kit) including (i) a first container containing an LSD1 inhibitor, (ii) a second container containing a further anti-cancer agent (as described above), and (iii) optionally, one or more further containers containing one or more further anti-cancer agents for use in the treatment of MPNST.

[0101] The manufactured article may further include a label or package insert. The term "package insert" refers to the instructions customarily included in the package of a commercially available therapeutic product and containing information about indications, usage, dosage, administration, contraindications, and / or warnings regarding the use of such therapeutic product. Suitable containers are, for example, blister packs, bottles, vials, syringes, etc. The container can be formed from various materials such as glass or plastic. The container can hold a composition or formulation effective in treating a disease and can also be provided with a sterile access port (for example, the container can be a bag or vial for intravenous solutions having a stopper penetrable by a hypodermic needle). The label or package insert indicates that this composition is used for the treatment of a selected disease, particularly MPNST. Alternatively or additionally, the manufactured article can further include a second container containing a pharmaceutically acceptable buffer such as bacteriostatic water for injection (BWFI), phosphate buffered saline, Ringer's solution, and dextrose solution. Further, other materials desirable from a commercial and user perspective, such as other buffers, diluents, filters, needles, and syringes, may be included.

[0102] The manufactured article or kit may further include instructions regarding the co-administration of one or more additional anti-cancer agents (as described above). For example, if the kit includes a first pharmaceutical composition / formulation containing an LSD1 inhibitor and a second pharmaceutical composition / formulation containing an additional anti-cancer agent, the kit may further include instructions regarding the simultaneous, sequential, or separate administration of the first and second pharmaceutical compositions / formulations to a subject in need thereof.

[0103] In another aspect, the manufactured article is suitable for the delivery of a solid oral dosage form of an LSD1 inhibitor, such as a tablet or capsule. Such manufactured articles preferably contain several dosage units. Such manufactured articles may include a card having dosage forms arranged in the order of their intended use. An example of such a manufactured article is a "blister pack". Blister packs are well known in the packaging industry and are widely used for packaging pharmaceutical dosage units. If desired, the day of the treatment schedule on which the dosage form can be administered can also be specified in the form of numbers, letters, or other marks or using a calendar fold-out to assist in memory.

[0104] According to one aspect, the manufactured article or kit may include: (i) a first container containing an LSD1 inhibitor; (ii) a second container containing a further anti-cancer agent; and optionally (iii) a third container containing a further anti-cancer agent. In this case, the anti-cancer agent in the third container is different from the anti-cancer agent in the second container. Alternatively, or additionally, the kit may include another container containing a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate buffered saline, Ringer's solution, and dextrose solution. Further, other materials that are desirable from a commercial and user perspective, such as other buffers, diluents, filters, needles, and / or syringes, may be included.

[0105] When the manufactured article or kit includes a composition of an LSD1 inhibitor and a composition of a further anti-cancer agent, the kit may include containers for containing the separate compositions, such as divided bottles or divided foil packets. However, the separate compositions may be contained within a single undivided container. Typically, the kit includes instructions for administering the separate components. The kit form is particularly beneficial when the separate components are preferably administered in different dosage forms (e.g., oral and parenteral), when they are administered at different dosing intervals, or when titration of the individual components of the combination is desired by the prescribing physician or veterinarian.

[0106] Definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0107] The following definitions apply throughout this specification and the claims, unless otherwise specified. For the purposes of the present invention, "subject" (or "patient") includes both humans and other animals, particularly mammals. Accordingly, the methods and uses of the present invention are applicable to both human therapy and veterinary applications. In a preferred embodiment, the subject (or patient) is a mammal (e.g., a human or a non-human mammal), most preferably the subject is a human (e.g., a male or female human). The human subject may be of any age, including, for example, 0 - 2 years old, 2 - 12 years old, 12 - 18 years old, or 18 years old and above.

[0108] The terms "treatment", "treating", etc. are generally used herein to mean obtaining the desired pharmacological and / or physiological effect. This includes partial or complete cure or amelioration of a disease (i.e., MPNST) and / or the symptoms or adverse effects resulting from the disease, or partial or complete arrest of the progression of the disease and / or the symptoms or adverse effects resulting from the disease. The term "treatment" as used herein encompasses any treatment of a disease (i.e., MPNST) in a subject. For example, it includes, but is not limited to, inhibition of MPNST, i.e., arrest, delay or slowdown of its onset / progression; or alleviation of MPNST, i.e., bringing about its (complete or partial) regression, remission, correction or reduction. The present invention relates specifically and expressly to each of these forms of treatment.

[0109] As used herein, the term "therapeutically effective amount" or "effective amount" of a compound according to the invention (in particular an LSD1 inhibitor) refers to an amount sufficient to produce a desired biological effect (e.g., a therapeutic effect or benefit) in a subject. Thus, a therapeutically effective amount of a compound may be an amount sufficient to treat a disease (i.e., MPNST), and / or delay the onset or progression of the disease, and / or alleviate one or more symptoms of the disease, when administered to a subject afflicted with or susceptible to the disease. The therapeutically effective amount will vary depending on the compound, the state of the disease being treated, the severity of the disease being treated, the age and relative health of the subject, the route and form of administration, the judgment of the attending physician or veterinarian, and other factors.

[0110] The term "pharmaceutically acceptable" means the properties of a material that is useful in the manufacture of a pharmaceutical composition, is generally safe, non-toxic, and neither biologically nor otherwise harmful, and is acceptable for veterinary and / or human pharmaceutical use.

[0111] As used herein, "pharmaceutically acceptable salts" are meant to include salts of the free acids and / or bases of particular compounds that retain the biological effectiveness of the specific compounds and are not biologically or otherwise harmful. Since compounds may have one or more sufficiently acidic or sufficiently basic functional groups, or both, they can react with several inorganic or organic bases, and either inorganic or organic acids, to form pharmaceutically acceptable salts. Exemplary pharmaceutically acceptable salts include salts produced by reacting the compounds described herein (especially LSD1 inhibitors, such as iadademstat) with a mineral or organic acid, such as hydrochloride, hydrobromide, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, nitrate, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caproate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, gamma-hydroxybutyrate, glycolate, tartrate, methanesulfonate (or mesylate), ethanesulfonate, propanesulfonate, benzenesulfonate (or besylate), toluenesulfonate, trifluoromethanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, pyruvate, stearate, ascorbate, or salicylate. When the compound (especially an LSD1 inhibitor) has an acidic moiety, suitable pharmaceutically acceptable salts can be salts formed with an alkali metal salt, such as sodium or potassium salt; an alkaline earth metal salt, such as calcium or magnesium salt; and suitable organic ligands such as ammonia, alkylamine, hydroxyalkylamine, lysine, arginine, N-methylglucamine, procaine, etc.Pharmaceutically acceptable salts are well known in the art (see, e.g., Stahl PH & Wermuth CG (eds.), “Handbook of Pharmaceutical Salts: Properties, Selection, and Use”, Wiley-VCH, 2002, and references cited therein, all of which are incorporated herein by reference).

[0112] The terms “pharmaceutical composition” and “pharmaceutical formulation” (or “formulation”) are used interchangeably and mean a mixture or solution containing a therapeutically effective amount of an active pharmaceutical ingredient (pharmaceutically active ingredient) (especially an LSD1 inhibitor) together with one or several pharmaceutically acceptable excipients, which is administered to a subject (e.g., a human) in need thereof.

[0113] The terms “pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier” can be used interchangeably and refer to any pharmaceutically acceptable component in a pharmaceutical composition that is non-toxic to the subject being administered and has no therapeutic activity. For example, disintegrants, binders, fillers, solvents, buffers, isotonic agents, stabilizers, antioxidants, surfactants, carriers, diluents, lubricants, etc. used in the formulation of pharmaceuticals. They are generally safe for human administration according to established government standards, including those promulgated by the US Food and Drug Administration and / or the European Medicines Agency. Pharmaceutically acceptable carriers or excipients are well known to those skilled in the art.

[0114] As used herein, the term “inhibitor” refers to a compound that competes with, reduces, blocks, inhibits, abrogates, or interferes with, in some way, the binding of a specific ligand to a specific receptor or enzyme, and / or that reduces, blocks, inhibits, abrogates, or interferes with, in some way, the activity or function of a specific protein, such as a receptor or enzyme.

[0115] As used herein, “small molecule” refers to an organic compound having a molecular weight of 900 Da (dalton) or less, preferably less than 500 Da. Molecular weight is the mass of a molecule and is calculated by multiplying the atomic weight of each constituent element in the molecular formula by the number of atoms of that element.

[0116] In this specification, the term "comprising" (or "comprise", "comprises", "contain", "contains", or "containing") has the meaning of "including, among other things", that is, "including... among any additional elements", unless otherwise specified or the context dictates otherwise. In addition to this, this term also includes the narrower meanings of "consisting essentially of" and "consisting of". For example, the term "A comprising B and C" has the meaning of "A including, among other things, B and C", and A can also contain any additional elements (for example, "A containing B, C, and D" is also included), but this term also includes the meanings of "A consisting essentially of B and C" and "A consisting of B and C" (that is, other components other than B and C are not included in A).

[0117] In this specification, the indefinite articles "a" and "an" and the definite article "the" include not only singular referents but also plural referents, unless the context clearly dictates otherwise.

[0118] The term "about" or "approximately" means an acceptable error with respect to a particular value determined by one of ordinary skill in the art. The error depends in part on how the value is measured or determined. In certain embodiments, the term "about" or "approximately" means within 1, 2, 3, or 4 standard deviations. In certain embodiments, the term "about" or "approximately" means within 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, or 0.05% of a given value or range. Any reference to a numerical value or range provided with the term "about" also includes a reference to the corresponding specific value or range.

[0119] Furthermore, whenever a numerical range is provided / described herein, it should be understood that all values and sub-ranges subsumed within each numerical range are specifically provided by the present invention. Accordingly, the present invention specifically and individually relates to each value falling within the numerical ranges described herein as well as to each and every sub-range subsumed by the numerical ranges described herein.

[0120] In this specification, various compounds are described by their chemical formulas and their corresponding chemical names. If there is any discrepancy between any of the chemical formulas shown herein and the corresponding chemical names, the present invention specifically and individually relates to the compounds defined by the chemical formulas and the compounds defined by the chemical names.

[0121] All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety.

Examples

[0122] The following examples are provided to illustrate the present invention. These should not be regarded as limiting the scope of the present invention, but should merely be regarded as representative of the present invention. Example 1: Effect of LSD1 inhibitor in MPNST viability assay 1.1 Experimental Design 50 μl / well of complete medium (1X GlutaMAX) in a 96-well plate TM, Mycoplasma-free MPNST cell lines (see Table 1) were seeded at an optimal cell density to ensure logarithmic growth throughout the experimental period in DMEM high glucose supplemented with 1 mM sodium pyruvate and 10% fetal bovine serum (FBS, Thermo Fisher). On the day after seeding, 50 μl of medium containing nine serial dilutions (1:3) of 2-fold concentrated iadademstat (used as the dihydrochloride salt, an LSD1 inhibitor) was added to the cells to obtain 100 μl of cells treated with the 1-fold concentrated compound at each dilution concentration. Etoposide (a topoisomerase II / DNA synthesis inhibitor) was also included as a positive control for the assay. Furthermore, two cell lines (sNF96.2 and sNF02.2) were selected from the MPNST cell line panel to evaluate the effects of two other LSD1 inhibitors, namely bomedemstat (bis tosylate salt) and pulrodemstat (besylate salt). Each experimental condition was tested in triplicate technical replicates, including medium-only wells and vehicle-treated controls for background correction and normalization, respectively. After treatment, the cells were incubated at 37 °C for 72 h in a humidified and controlled 5% CO 2 atmosphere. At this point, compound and medium refreshment was performed by adding 50 μl of medium supplemented with the 1-fold concentrated compound at each corresponding dilution concentration. After the cells were incubated for an additional 72 h (for a total of 6 days), cell viability was evaluated using either the MTT assay (Sigma-Aldrich) or AlamarBlue TM Cell Viability Reagent (Life Technologies) according to the manufacturer's instructions. The background was calculated as the mean of the medium-only control values and subtracted from each data point. The mean of the background-corrected technical replicates was calculated and normalized by the mean of the vehicle-treated control (corresponding to 100% viability). The data were analyzed using GraphPad PRISM® version 9.0.1 (GraphPad Software, Inc., La Jolla, CA, USA) to generate the best-fit curve and EC 50 values (corresponding to the concentration of the compound at which half (50%) of the maximum effect is obtained; thus, the lower the EC 50 value, the stronger the potency).

[0123]

Table 1

[0124] 1.2 Results Using a panel of 12 MPNST cell lines, the effect of the LSD1 inhibitor iadademstat on the viability of MPNST cells was evaluated after 6 days of treatment. Etoposide was used as a positive control for the assay. To reliably reflect in vitro the clinical situation where MPNST patients are characterized by high genomic heterogeneity, a panel of different types of MPNST cell lines was used. The panel included both MPNST immortalized cell lines and patient-derived MPNST cell lines. Furthermore, some MPNST cell lines were type I neurofibromatosis-related and others were sporadic, and in both cases there were variations in their genomic backgrounds with respect to the mutational status of NF1, CDKN2A, and PRC2. An overview of their characteristics is provided in Table 2 below.

[0125]

Table 2

[0126] * The patient-derived MPNST-NF1-18b cell line was initially classified as a sporadic MPNST cell line and named MPNST-SP-09b. In subsequent analysis, this cell line was confirmed as a type I neurofibromatosis-related MPNST cell line, so its name was changed to MPNST-NF1-18b.

[0127] LSD1 inhibitors such as iadademstat have been reported to exert a therapeutic effect not by killing cancer cells, but by inducing cancer cell differentiation and inhibiting cancer cell proliferation (Sacilotto N et al., ACS Pharmacol Transl Sci, 2021, 4(6):1818 - 34, doi:10.1021 / acsptsci.1c00223). Accordingly, a decrease in cancer cell survival rate exceeding 30% reflects the strong therapeutic effect of the corresponding LSD1 inhibitor. Therefore, in this experiment, the responses obtained after treatment with LSD1 inhibitors were classified into the following three groups. (1) Strong response (survival rate decrease > 30%); (2) Moderate response (survival rate decrease > 15% and < 30% (more than 15% and less than 30%)); (3) Low response (survival rate decrease < 15%). Table 3 shows an overview of the classification by the decrease in survival rate after treatment with LSD1 inhibitors, together with the EC 50 values obtained with etoposide (positive control of the assay) and iadademstat.

[0128]

Table 3

[0129] Despite the large genomic and mutational variations within the tested MPNST cell line panel, 8 out of 12 cell lines showed a particularly beneficial and good response to LSD1 inhibitor treatment. Therefore, after 6 days of treatment with iadademstat, 3 out of 12 cell lines (25%) showed a strong decrease in survival rate, and 5 out of 12 cell lines (41.67%) showed a moderate decrease in survival rate. Notably, the particularly responsive cell lines included both neurofibromatosis type I - related MPNST and sporadic (not neurofibromatosis type I - related) MPNST cell lines, indicating that treatment with LSD1 inhibitors (such as iadademstat) exerts a therapeutic effect on a wide range of MPNST patient subsets, including neurofibromatosis type I - related MPNST as well as non - neurofibromatosis type I - related MPNST. Furthermore, in both moderately responsive and strongly responsive cell lines, which accounted for a total of 66.67%, iadademstat had an EC 50It shows the values (see Table 3), indicating that there is a clinically significant therapeutic effect even when administered at very low doses.

[0130] The effects of LSD1 inhibitors on the cell viability of MPNST were further tested using two additional LSD1 inhibitors, namely bomedemstat and pulrodemstat. Bomedemstat is an irreversible LSD1 inhibitor like iadademstat, while pulrodemstat is a reversible LSD1 inhibitor. Cell viability was evaluated after 6 days of treatment in both the sNF96.2 and sNF02.2 cell lines as per the method described above (see Table 4). The results thus obtained, as presented in Table 4 (and Table 3 above for iadademstat), clearly show that all three LSD1 inhibitors, whether irreversible or reversible, are effective against MPNST.

[0131]

Table 4

[0132] Among the LSD1 inhibitors tested, iadademstat showed the highest potency (lowest EC 50 ) in both the sNF96.2 and sNF02.2 cell lines. These results indicate that LSD1 inhibitors, including both irreversible (e.g., iadademstat and bomedemstat) and reversible (e.g., pulrodemstat) LSD1 inhibitors, are beneficially effective in the treatment of MPNST and can be used for a wide range of different subgroups of MPNST patients.

[0133] Example 2: Matrix assay to determine synergistic effect between LSD1 inhibitor and MEK inhibitor in MPNST cell lines 2.1 Experimental design 2.1.1 Matrix viability assay (6 days) Each matrix assay was assigned to either 2 plates in a 9×9 format according to the scheme shown in Figure 1 or 1 plate in a 5×5 format according to the scheme shown in Figure 2. The iadademstat, an LSD1 inhibitor, was added while increasing the concentration from top to bottom, and the selumetinib, a MEK inhibitor, was added while increasing the concentration from left to right.

[0134] For the assay, cells were seeded at the optimal density specified in Example 1 in 50 μL of medium in a 96-well plate. The wells at the edges of the plate were filled with 100 μL of medium only for background correction. Each of the two compounds to be combined was added to 25 μL at a 4-fold concentration, with a final volume of 100 μL and a final concentration of 1-fold for each dilution (DMSO% < 0.5%). In the 9×9 matrix, the compounds were added at a 1:2 serial dilution, while in the 5×5 matrix, the compounds were added at a 1:5 serial dilution, both ensuring that the concentration range covered a complete dose-response curve. As shown in Figures 1 and 2, the matrix was designed such that the predicted EC 50 values of both compounds were placed at the center in the horizontal and vertical directions of the matrix. In this way, the wells on the diagonal of the plate corresponded to a fixed EC 50 ratio between the two compounds. In the 9×9 matrix, to confirm the reproducibility between the two plates, the first and last rows of Plate #1 were repeated in Plate #2 (indicated by arrows in Figure 1). The EC 50 values of the compounds tested in the matrix assay were previously obtained through single-agent assays conducted as detailed in Example 1 and are shown in Table 3 for iadademstat. After treatment, the cells were incubated at 37 °C for 72 hours in a humidified and controlled 5% CO 2 atmosphere. At this point, the compounds and the medium were refreshed by adding 50 μl of medium supplemented with 1-fold concentrated compound at each corresponding dilution concentration. After the cells were incubated for an additional 72 hours (for a total of 6 days), the cell viability was measured in two biological replicates (biological replicates) using the MTT assay (Sigma-Aldrich) or AlamarBlue according to the manufacturer's instructions. TMEvaluated using any of the cell viability reagents (Life Technologies). The background was calculated as the average of the control values of the medium only and subtracted from each data point. The background-corrected values were normalized by the corrected vehicle-treated control (corresponding to 100% viability). The data were analyzed using GraphPad PRISM® version 9.0.1 (GraphPad Software, Inc., La Jolla, CA, USA), and the best-fit curve and EC 50 value (corresponding to the concentration of the compound at which half (50%) of the maximum effect is obtained; thus, the lower the EC 50 value, the stronger the potency) were calculated.

[0135] 2.1.2 Matrix viability assay (data analysis) For each matrix assay, the data were normalized against the vehicle-treated control (<0.5% DMSO, upper left corner), and the percentage value of relative residual viability was obtained according to the following formula.

[0136] % Relative residual viability = Background-corrected signal of treated cells / Background-corrected signal of vehicle control × 100 The percentage values of residual viability were then analyzed using GraphPad PRISM® version 9.0.1 (GraphPad Software, Inc., La Jolla, CA, USA), and the best-fit curve and EC 50 value of the single agent were calculated.

[0137] At this point, the fraction affected (Fa) (also known as fractional effect) was calculated for the following conditions using the following formula. Fa = 1 - (% relative residual viability / 100) · Cells treated with serial dilutions of selumetinib as a single agent.

[0138] · Cells treated with serial dilutions of iadademstat as a single agent. · EC 50Cells treated with iadademstat and selumetinib at a fixed ratio corresponding to the ratio of values (% relative survival values on the diagonal of the matrix assay; highlighted in Figures 1 and 2).

[0139] CalcuSyn software (http: / / www.biosoft.com / w / calcusyn.htm, Biosoft, Cambridge, UK) is designed to determine the nature (synergistic, additive, or antagonistic) of the interaction between two compounds by calculating the combination index (CI). This analysis is based on the Median Effect Principle and the Combination Index Theorem as described by the Chou-Talalay method (Chou TC, Pharmacol Rev, 2006, 58(3):621-681, doi:10.1124 / pr.58.3.10). According to this, a resulting CI < 1 indicates a synergistic effect, CI = 1 indicates an additive effect, while CI > 1 reflects an antagonistic effect. In the case of a synergistic effect (CI < 1), the smaller the CI value, the greater the synergistic effect. Furthermore, the strength of the drug interaction can be further classified based on the CI range as shown in Table 5.

[0140]

Table 5

[0141] To generate useful and consistent results, it is necessary to fit the data (both single-agent and drug combinations) processed by CalcuSyn to the theoretical models of the Median Effect Principle and the Combination Index Theorem. For this reason, it is important to remove outliers and data points that are characterized as not fitting well to the Median Effect Principle (Chou TC, Pharmacol Rev, 2006, 58(3):621-681, doi:10.1124 / pr.58.3.10). To achieve this, the following strategies were adopted for data filtering.

[0142] As a first step, data dispersion was reduced by removing the points characterized as follows. 1) Fa < 0.1 2) An increase in Fa < 0.03 compared to the previous point (when Fa > 0.9).

[0143] These conditions define a plateau of the dose - response curve where the survival rate drops to near 0% or 100% (corresponding to Fa values close to 0 or 1, respectively) as a result of cells being treated with very low or very high concentrations of a compound (or combination). It should be noted that in these regions of the dose - response curve, the change in alamarBlue TM or MTT signal is very small and is likely due to random noise with little biological significance.

[0144] Next, for each data point, Log 10 (concentration) and Log 10 (Fa / (1 - Fa)) were calculated, and a dot - plot graph was generated reporting the former value on the x - axis and the latter value on the y - axis. Then, a regression line was obtained using Excel (corresponding to the median - effect equation).

[0145] At this point, the distance from the regression line for each data point was calculated using the following formula. Distance(ax + by + c = 0; X,Y)=(aX + bY + c) / √(a 2 + b 2 ) Outliers were identified using the Grubbs' test based on their distances from the median - effect equation. For each data point, the Grubbs' test was performed on the absolute value of the distance according to the following formula (note that the variable for the Grubbs' test can be called G or Z interchangeably).

[0146] G=(X n - X average ) / s In the formula, X n represents the absolute value of the distance of each point from the regression line; X average represents the mean of all X n values, and s represents the standard deviation. G critValues of G above (calculated with α = 0.2 as shown below) are identified as outliers that do not conform to the median effect equation. Such data points are removed so that the combination index by CalcuSyn can be successfully calculated.

[0147]

Number

[0148] If possible, 1. no further outliers are identified, or 2. the test is repeated more than twice until 2 R > 0.95

[0149] 2.1.3 CalcuSyn output results The results of CalcuSyn are for fixed EC 50The experimental fractional effect (referred to as Fa) representing the percentage of cells affected by combination treatment in ratio (in the case of cytotoxic treatment, the fractional effect corresponds to the reduction in survival rate compared to the vehicle control, and Fa = 1 is equal to a 100% reduction in survival rate) is provided along with the related combination index (CI). As shown in Table 5 above, the CI value is an indicator of the nature and strength of the interaction of the compounds. A value less than 1 indicates a synergistic interaction (the closer the value is to 0, the stronger the synergistic effect), a value equal to 1 indicates an additive interaction, and a value greater than 1 indicates an antagonistic interaction. The software also provides simulations of CI and Fa based on experimental data, and provides the estimated CI at ED75 and ED90 (the effective doses corresponding to 75% and 90% reduction in survival rate respectively). In the case of anti-cancer agents or anti-viral drugs, as described in Chou TC, Cancer Res (2010) 70(2):440-446., doi:10.1158 / 0008-5472.CAN-09-1947, a synergistic effect at a high effect level (e.g., Fa > 0.75) is more appropriate for treatment than at a low effect level (e.g., Fa < 0.2).

[0150] Data are provided as both the experimental CI for the relevant Fa for each experiment and the mean estimated CI for ED75 and ED90 obtained from two independent biological replicates. 2.2 Results Matrix treatment with the MEK inhibitor selumetinib and the covalent irreversible LSD1 inhibitor iadademstat was carried out as described in section 2.1.1. Data analysis and calculation of the combination index were carried out as described in section 2.1.2. The combination index (CI) related to the specific fractional effect (Fa), the estimated CI at ED75 and ED90, and the results of their respective classifications (described in Table 5) obtained from the combination of iadademstat and selumetinib are shown in Tables 6 and 7.

[0151] In summary, the combination of iadademstat and selumetinib showed a strong synergistic effect in three tested cell lines, including the patient-derived MPNST-NF1-18b cell line, at a wide range of fractional effects (Fa). Importantly, these synergistic effects were observed at biologically meaningful effective doses (ED75 and ED90).

[0152]

Table 6

[0153]

Table 7

[0154] These results indicate that the LSD1 inhibitor (e.g., iadademstat) and the MEK inhibitor (e.g., selumetinib) interact synergistically in a panel of various MPNST cell lines, and the combination of these agents is particularly beneficial for the treatment of MPNST.

[0155] Example 3: Matrix assay to determine synergistic effect between LSD1 inhibitor and Pi3K inhibitor in MPNST cell lines 3.1 Experimental Design Matrix viability assays and corresponding analyses were performed as described in Example 2, except that the LSD1 inhibitor iadademstat and the PI3K inhibitor copanlisib were used.

[0156] 3.2 Results Matrix treatment with the PI3K inhibitor copanlisib and the covalent irreversible LSD1 inhibitor iadademstat was performed as described in section 2.1.1. Data analysis and calculation of the combination coefficient were performed as described in section 2.1.2. The combination coefficients (CI) related to the specific fractional effect (Fa), the estimated CI at ED75 and ED90, and the results of each classification (described in Table 5) obtained from the combination of iadademstat and copanlisib are shown in Tables 8 and 9.

[0157] In summary, the combination of iadademstat + copanlisib showed a synergistic effect across a wide range of response rates (Fa). Importantly, these synergistic effects were observed at biologically meaningful effective doses (ED75 and ED90).

[0158]

Table 8

[0159]

Table 9

[0160] These results indicate that the LSD1 inhibitor (such as iadademstat) and the Pi3K inhibitor (such as copanlisib) interact synergistically in various MPNST cell lines, and that the combination of such agents is particularly beneficial for the treatment of MPNST.

[0161] Although the invention has been described in connection with its specific embodiments, it will be appreciated that further modifications are possible, that this patent or this patent application generally covers any variations, uses or adaptations of the invention that follow the principles of the invention, and that departures from the present disclosure that fall within the scope of known or customary practice in the technical field to which the invention pertains and that are applicable to the essential features described herein and that follow the appended claims are also included.

Claims

1. A pharmaceutical composition for use in the treatment of malignant peripheral nerve schwannoma (MPNST), comprising an LSD1 inhibitor and one or more pharmaceutically acceptable excipients.

2. Use of LSD1 inhibitors for the manufacture of pharmaceutical compositions for the treatment of malignant peripheral nerve schwannoma (MPNST).

3. The pharmaceutical composition according to claim 1, wherein the LSD1 inhibitor is a small molecule.

4. The pharmaceutical composition according to claim 1, wherein the LSD1 inhibitor is selected from the group consisting of iademstat, plurodemstat, bomedemstat, secridemstat, 1-((4-(methoxymethyl)-4-(((1R,2S)-2-phenylcyclopropylamino)methyl)piperidine-1-yl)methyl)cyclobutancarboxylic acid, 3-(cyanomethyl)-3-(4-{[(1R,2S)-2-phenylcyclopropyl]amino}piperidine-1-yl)azetidine-1-sulfonamide, 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]phenyl]-2-fluorobenzonitrile, and pharmaceutically acceptable salts thereof.

5. The pharmaceutical composition according to claim 1, wherein the LSD1 inhibitor is selected from the group consisting of idademstat, prurodemstat, bomedemstat, and pharmaceutically acceptable salts thereof.

6. The pharmaceutical composition according to claim 1, wherein the LSD1 inhibitor is idademstat or a pharmaceutically acceptable salt thereof.

7. The pharmaceutical composition according to claim 6, wherein the LSD1 inhibitor is idademstat dihydrochloride.

8. The pharmaceutical composition according to any one of claims 1 or 3 to 7, wherein MPNST is neurofibromatosis type 1-associated MPNST, sporadic MPNST, or radiation-induced MPNST.

9. The pharmaceutical composition according to any one of claims 1 or 3 to 7, wherein the pharmaceutical composition is administered to a subject having neurofibromatosis type 1.

10. The pharmaceutical composition according to any one of claims 1 or 3 to 7, wherein the pharmaceutical composition is administered to a subject who does not have neurofibromatosis type 1.

11. The pharmaceutical composition according to any one of claims 1 or 3 to 7, wherein MPNST has one or more mutations or genetic changes that affect the NF1 gene.

12. The pharmaceutical composition according to any one of claims 1 or 3 to 7, wherein MPNST has one or more mutations or genetic alterations that affect the expression and / or activity of CDKN2A, p53, RB1, PTEN, PRC2, EGFR, PDGFRA, and / or c-Met.

13. The pharmaceutical composition according to any one of claims 1 or 3 to 7, wherein MPNST is a transferable MPNST.

14. The pharmaceutical composition according to any one of claims 1 or 3 to 7, wherein the pharmaceutical composition is administered to a human subject.

15. The pharmaceutical composition according to any one of claims 1 or 3 to 7, wherein the pharmaceutical composition is administered orally.

16. The pharmaceutical composition according to any one of claims 1 or 3 to 7, wherein the pharmaceutical composition is administered in combination with one or more further anticancer agents and / or in combination with radiotherapy.

17. The use according to claim 2, wherein the LSD1 inhibitor is selected from the group consisting of iademstat, prurodemstat, bomedemstat, and pharmaceutically acceptable salts thereof.

18. The use according to claim 2, wherein the LSD1 inhibitor is idademstat or a pharmaceutically acceptable salt thereof.

19. The use according to claim 2, wherein the LSD1 inhibitor is idademstat dihydrochloride.

20. The use according to any one of claims 2 or 17 to 19, wherein MPNST is neurofibromatosis type 1-associated MPNST, sporadic MPNST, or radiation-induced MPNST.

21. The use according to any one of claims 2 or 17 to 19, wherein MPNST has one or more mutations or genetic alterations that affect the NF1 gene.

22. A manufactured article for use in the treatment of malignant peripheral nerve schwannoma (MPNST), comprising an LSD1 inhibitor and one or more additional anticancer agents in the same or another pharmaceutical formulation.

23. - The LSD1 inhibitor is as defined in any one of claims 3 to 7; and / or - MPNST is as defined in any one of claims 8 or 11-13; and / or - The subject to whom the manufactured article is administered is as defined in any one of claims 9, 10, or 14; and / or - The manufactured product is administered orally. The manufactured article according to claim 22.

24. The manufactured article according to claim 22, wherein one or more further anticancer agents include a MEK inhibitor.

25. The pharmaceutical composition according to any one of claims 1 or 3 to 7, wherein the pharmaceutical composition is administered in combination with a MEK inhibitor.

26. The pharmaceutical composition according to claim 25, wherein the MEK inhibitor is selected from selumetinib, trametinib, cobimetinib, binimetinib, mildametinib, pimacertib, refametinib, zapnometinib, abutometinib, HL-085, FCN-159, TAK-733, and pharmaceutically acceptable salts thereof.

27. The pharmaceutical composition according to claim 25, wherein the MEK inhibitor is selumetinib or a pharmaceutically acceptable salt thereof, and the LSD1 inhibitor is idademstat or a pharmaceutically acceptable salt thereof.

28. The manufactured article according to claim 22, wherein one or more further anticancer agents include a Pi3K inhibitor.

29. The pharmaceutical composition according to any one of claims 1 or 3 to 7, wherein the pharmaceutical composition is administered in combination with a Pi3K inhibitor.

30. The pharmaceutical composition according to claim 29, wherein the Pi3K inhibitor is selected from copanlisib, alpelisib, idelalisib, duvelisib, umbralicib, buparlicib, zandelisib, limperlisib, palsacricib, reniolisib, paxalisib, inavolisib, cerabelisib, pictilisib, taselicib, tenalisib, eganerisib, GSK2636771, MEN1611, AMG-319, and pharmaceutically acceptable salts thereof.

31. The pharmaceutical composition according to claim 29, wherein the Pi3K inhibitor is copanlisib or a pharmaceutically acceptable salt thereof, and the LSD1 inhibitor is idademstat or a pharmaceutically acceptable salt thereof.

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