Malt-1 inhibitor for treating malt-1 mediated diseases

Abstract

The present invention relates to gliptins, the inhibitors of dipeptidyl peptidase 4, also known as DPP-4 inhibitors, for use in the treatment and / or prevention of diseases in a human subject. The present invention relates to a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating a MALT-1 involving disease in a subject, preferably selected from allergic inflammatory disease, an autoimmune disease, multiple myeloma and glioma, in a human subject in need thereof. The present invention also relates to the use of gliptins as an allosteric MALT1 inhibitor for use in a method of treating a MALT-1 involving disease in a subject, preferably selected from an allergic inflammation disease, multiple sclerosis, multiple myeloma and glioma. The present invention is also directed to the field of personalized cancer care and treatment.

Description

[0001] MALT-1 INHIBITOR FOR TREATING MALT-1 MEDIATED DISEASES

[0002] TECHNOLOGICAL FIELD

[0003] The present invention relates to gliptins, the inhibitors of dipeptidyl peptidase 4, also known as DPP-4 inhibitors, for use in the treatment and / or prevention of (MALT-1 mediated or MALT-1 involving) diseases in a human subject. The present invention is also directed to the field of personalized cancer care and treatment. In particular, a targeted approach is provided herein for treating neurological (e.g. brain) cancers (over)expressing mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1), with inhibitors of dipeptidyl peptidase-4 (DPP-4), also known as gliptins. The present invention relates to a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating a MALT-1 mediated or involving disease in a subject, preferably selected from allergic inflammatory disease, an autoimmune disease, preferably multiple sclerosis, multiple myeloma (disease) and glioma, in a subject, preferably human subject, in need thereof. The method further relates to an oral administration of said pharmaceutical composition, wherein the method results in inhibition of intracellular MALT1 activity, preferably inside aberrant cells such as for example autoimmune cells (e.g. in a patient suffering from MS), tumor cells, such as for example glioma cells or myeloma cells, or cells involved in allergic inflammatory disease. The present invention also relates to the use of gliptins as an allosteric MALT1 inhibitor for use in a method of treating a MALT-1 mediated or involving disease in a subject, preferably selected from an allergic inflammation disease, an autoimmune disease, preferably multiple sclerosis, multiple myeloma and glioma. The present invention further relates to a use of gliptins in the manufacture of a medicament for the therapeutic or prophylactic treatment of a MALT-1 mediated or involving disease in a subject, preferably selected from allergic inflammatory disease, an autoimmune disease, preferably multiple sclerosis, multiple myeloma (disease) and glioma, in a subject, preferably human subject, in need thereof. Further provided herein are in vitro diagnostic methods for determining an increased probability that a neurological cancer of a human subject will respond to the treatment with a gliptin, the methods comprising determination of MALT1 expression status and, optionally, of DPP-4 expression status in a sample obtained from the subject.

[0004] BACKGROUND

[0005] ALLERGIC DISORDERS

[0006] Allergic disorders, such as anaphylaxis, hay fever, eczema and asthma, now afflict roughly 25% of people in the developed world. In allergic subjects, persistent or repetitive exposure to allergens, which typically are intrinsically innocuous substances common in the environment, results in chronic allergicinflammation. This in turn produces long-term changes in the structure of the affected organs and substantial abnormalities in their function. (Galli 2008).

[0007] In the past decade, research in the molecular and cellular underpinnings of basic and clinical immunology has significantly advanced the understanding of allergic disorders, allowing scientists and clinicians to diagnose and treat disorders such as asthma, allergic and nonallergic rhinitis, and food allergy.

[0008] Certain key regulatory cytokines, genes and molecules have recently been shown to play key roles in allergic disorders. For example, interleukin-33 (IL-33) plays a (an important) role in refractory disorders such as asthma, allergic rhinitis and food allergy, mainly by inducing T helper (Th) 2 immune responses (Han 2020).

[0009] New studies on allergic diseases are underway, especially the development and application of new biologies. So far, many biologies targeting Th2 / 1 / 17 inflammatory biomarkers are available, many of which are clinically applied. Identifying new and reliable biomarkers and clarifying novel molecular mechanisms that persist in specific reactions have become important research directions.

[0010] Studies using both human and mouse model systems have provided key evidence for the role that Th2 cytokines play in driving many of the hallmarks of allergic inflammation. Furthermore, the signalling pathways by which Th2 cytokines exert their effects on airway target cells are rapidly being elucidated, and antagonists of the Th2 pathway are under active development (Georas 2005).

[0011] IL-4 and IL-13 represent key driving factors in type 2 immune diseases. A type 2 immune response typically happens when your body senses certain kinds of infections or allergens and sends out specific type 2 immune cells to fight a battle against the intruders by creating inflammation. Type 2 immune diseases include among others Asthma, Atopic Dermatitis, Nasal polyps, eosinophilic esophagitis, prurigo nodularis and certain food allergies (Wang 2023).

[0012] Current treatments of allergic inflammatory disease are still limited or not adequate. The pathogenesis of allergic diseases is complex, involving many factors such as genetics, epigenetics, environmental factors, microecology and the body’s immune function. Their recurrence rate is high, which brings great pain to and imposes a severe financial burden on patients.

[0013] To conclude, there is still a need for improved treatments for allergic inflammatory patients or for management of allergic inflammatory diseases responsive to different episodes and symptoms of the disease.

[0014] AUTOIMMUNE DISEASE

[0015] Multiple sclerosis (MS) is an autoimmune central nervous system (CNS) disorder that causes demyelination and neuronal injury. MS is one of the most common causes of non-traumatic disability among young adults (aged 18-40 years). MS is a heterogenous, multifactorial and immune-mediated disease which is caused by complex gene-environment interactions.

[0016] According to the latest Multiple Sclerosis Atlas, a joint project between the Multiple Sclerosis International Federation and WHO, 2.8 million have multiple sclerosis worldwide. Approximately 900.000 people are affected by MS in the US. The prevalence of MS worldwide ranges from 5 to 300per 100.000 people and increases in higher latitudes. Latitude effects can be correlated with higher exposure to sun which increases vitamin D levels, which are associated with a lower prevalence of MS.

[0017] Genome-wide association studies have identified more than 200 genetic risk variants associated with multiple sclerosis susceptibility. Most of these variants involve pathways of peripheral immune cells and resident microglia, with the strongest association with polymorphisms in HLA genes, in particular HLA-DRB1*15:01. The genomic map provides evidence that multiple sclerosis is primarily a neuroinflammatory disorder and not primarily a neurodegenerative disorder that is complicated by superimposed inflammatory events. Risk genes for multiple sclerosis do not overlap with those for progression of multiple sclerosis or those of other neurodegenerative disorders. International Multiple Sclerosis Genetics Consortium elucidates how heritable contributions affects individual phenotypes and prognosis. A recent, genome-wide association study showed that presence of rs10191329 in the DYSF-ZNF638locus confers faster disease progression and is associated with greater cortical pathology.

[0018] MS is characterized by inflammatory demyelination and axonal transection, defined as severed terminal axonal structures representing the pathological correlate of irreversible neurologic damage. The accumulation of demyelinating lesions that occur in the white matter and the grey matter of the brain and spinal cord.

[0019] In most patients, reversible episodes of neurological deficits, also called relapses, usually last for days or weeks and characterize the initial phases of the disease. Four clinical courses of MS have been identified: relapsing-remitting MS (RRMS), secondary progressive MS (SPMS), primary progressive MS (PPMS) and progressive relapsing MS (PRMS). Approximately 85% of the patients display RRMS which is characterized by the occurrence of relapses at irregular intervals with complete or incomplete neurological recovery. Most patients with RRMS develop also SPMS which is characterized by which is characterize by progressive, irreversible disability that occurs independently of the presence of relapses. Almost 10-15% of patients present with PPMS, another course of MS, characterized by disease progression from the onset, resulting in gradual, progressive and permanent neurological deficits for >1 year without relapses. A rare clinical course is PRMS characterized by progressive disease from the onset, with acute relapses (with or without full clinical recovery) and periods of continuing progression between relapses. A further phenotype includes clinically isolated syndrome (CIS), denoting the condition of patients whose first clinical presentation has characteristics of inflammatory demyelination that could be MS but who do not fulfil its diagnostic criteria. Within each subtype, it is possible to diagnose and classify the disease whether it is active or not active which are defined by the occurrence of relapses or lesions detected using MRL Another important modifier of the progressive stages is the inclusion of whether disability has progressed over a given time period.

[0020] The pathology seen in all MS phenotypes is the formation of focal plaques also known as lesions which are areas of demyelination that ae typically located around post-capillary venules and are characterized by breakdown of the blood-brain barrier (BBB). The mechanism of said breakdown is not fully understood but seems to involve direct effects of pro-inflammatory cytokines and chemokines such as TNF, IL-1 B and IL-6 produced by resident cells, endothelial cells. Due to the dysregulation of the BBB, the trans-endothelial migration of activated leukocytes including macrophages, T cells and B cells,into the CNS increases, which leads to further inflammation and demyelination, followed by oligodendrocyte loss, reactive gliosis and neuro-axonal degeneration.

[0021] Current treatments of MS are still limited and involve lifelong use of disease-modifying agents to shorten duration of acute exacerbations, decrease their frequency, and provide symptomatic relief. All currently available treatments display one of more significant adverse effect(s), including injection site reactions, infections, liver damage, increased blood pressure, reduced heart rate, macular oedema, skin cancer, leukopenia and autoimmune reactions. Symptomatic treatments are aimed at maintaining function and improving quality of life. Some of these symptoms include spasticity, pain, fatigue, cognitive impairment, bladder and bowel issues, gait dysfunction, mood dysregulation, and sleep disturbance. Treating these symptoms should include a combination of pharmacological and non-pharmacological treatments.

[0022] There still exists a need for new treatments for MS patients.

[0023] CANCER, IN PARTICULAR NEUROLOGICAL CANCERS SUCH AS GLIOMA

[0024] Cancers are a large family of diseases characterized by uncontrolled growth of genomically-altered cells in the body. The vast complexity of cancer phenotypes and their underlying and dynamically -changing genotypes can be daunting in the breadth and scope of their diversity, cell and tissue biology, pathology, and responses to therapy (Hanahan 2022). For this reason, it is becoming generally recognized and accepted that every cancer is genetically unique, and that effectively treating these diseases will eventually rely on a personalized (or “precision”) medicine.

[0025] Personalized approach to cancer treatment takes into account the inter- and intra-tumor variability in genes, and their interactions with the surrounding tumor immune environment and within different expression patterns. In the cancer field, precision medicine is particularly promising from the patient’s safety perspective, as it aims to target specific oncogenic drivers without interfering with the healthy processes within the body and without triggering the cancer’s evolution towards more aggressive forms through inadvertent stimulation of other oncogenic pathways present within cancer cells. The ability to select tailor-made therapeutic interventions to which a given patient’s cancer will have an increased probability of responding has also recently become more practically feasible thanks to the increasing availability of affordable diagnostics, allowing personalized gene expression analyses and genetic patient-profiling.

[0026] Particularly difficult to treat forms of cancer include neurological cancers, which due to their hidden localization (in the skull or spine) and the general histological complexity of the nervous tissue, have very few well-characterized compound-targetable gene expression targets. Among different neurological cancers, gliomas are the most common primary brain cancers, encompassing a diverse group of tumors that arise from glial (i.e. non-neuronal) cells. Gliomas are estimated to account for approximately 30% of all brain tumors and 80% of all malignant brain tumors with high morbidity and mortality. High-grade (i.e. grade III or IV) malignant gliomas are referred to as glioblastomas, among which the most aggressive, lethal, and prevalent subtype, glioblastoma multiforme (GBM), is characterized by rapid growth, high degree of invasiveness, and substantial genetic heterogeneity withina single tumor mass. This extreme variability (as reflected in the cancer’s name “multiforme”, meaning “highly variable”) makes the selection of therapeutic interventions for GBM very difficult.

[0027] The usual treatment of glioblastoma includes maximal safe surgical resection followed by radiotherapy and concomitant temozolomide chemotherapy. In view of the high genetic variability of this cancer, many therapeutic approaches aiming to target glioblastoma at the molecular level have been proposed and are being tested, including small-molecule inhibitors, monoclonal antibodies, gene therapies, oncolytic viruses etc. Among them, in CN111920816, use of sitagliptin was proposed for specifically inhibiting dipeptidyl peptidase 4 (DPP-4 or DPP-IV) as a molecular target for treating GBM.

[0028] However, in the context of GBM and several other neurological cancers, in the absence of patient’s genetic profiling data evidencing pathological indicators pointing to the contrary (e.g. abnormal DPP-4 overexpression or gene locus amplification etc.), targeted inhibition of DPP-4 is highly controversial. Namely, a substantial number of research reports suggests that the DPP-4 may actually act as a tumor-suppressor, esp. in gliomas including glioblastomas (Sedo et al., 2008), and in neuroblastoma (Arscott et al., 2008), which would make its targeted inhibition by gliptins undesired.

[0029] DPP-4 is a key enzyme in regulating body sugar metabolism by cleaving and inactivating the two incretin hormones: glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP). Circulating DPP-4 concentration is known to be higher in individuals with obesity than in those with normal body weight (Rohmann et al., 2021). Forthose reasons, gliptins are widely used to treat type 2 diabetes. DPP4 also participates in inflammation, e.g. by cleaving chemokines and cytokines (Klemann et al., 2016; Wronkowitz et al., 2014), but despite its positive correlation with obesity and inflammation, the role of DPP-4 in cancer is not clear and appears to be tumor type dependent.

[0030] On one hand, DPP-4 expression in colorectal and some hematological cancers is high and appears to correlate with tumor progression (Sato et al., 2005; Okamoto et al., 2014). On the one hand, DPP-4 was shown to act as a tumor suppressor in other cancer types, such as melanoma, non-small cell lung carcinoma, prostate and ovarian cancers and, notably, neurological cancers including glioblastoma (Wesley et al., 1999; Kajiyama et al., 2002; Sedo et al., 2008). These differences in DPP-4 effects were hypothesized as resulting from its interactions with the tumor microenvironment (Pro and Dang, 2004) and location-dependent chemokine cleavage (Arscott et al., 2009).

[0031] For example, known DPP-4 substrates include several poly- and oligopeptides strongly associated with the malignant behavior of GBM cells, such as the chemokine stromal cell-derived factor (CXCL12, SDF-1) and the neuropeptide substance P (SP) (Mentlein, 1999; Bajetto et al., 2006; Palma and Maggi, 2000; Beckenkamp et al., 2016), which could perhaps explain why some neurological malignancies select for decreased or lost DPP-4 expression in order to attain more aggressive metastatic phenotype. Furthermore, DPP-4 has also been considered as a surface marker of senescent cells (Kim et al., 2017), and recently it was reported that DPP-4 inhibition could result in decreased production of the potent cell cycle inhibitor proteins p53 and p21 (Chen et al., 2020b).

[0032] As a consequence, the association between DPP-4 and malignancy is highly debated and in neurological cancers it appears to be on the tumor-suppressive side. In particular, in cultured glioma cells, expression of DPP-4 correlates with reduced growth, migration, and adhesion in vitro, and further, overexpression of DPP-4 suppressed glioma growth in an orthotopic xenotransplantation mouse modelin vivo (Busek et al., 2012). In another report, endogenous DPP-4 upregulation in response to serum withdrawal was reported to induce differentiation and growth arrest of astrocytic glioblastoma cells (Balaziova et al., 2011). In the childhood cancer neuroblastoma, which frequently overexpresses mitogenic peptides, chemokines and their receptors, DPP-4 was shown to inactivate or degrade some of these factors, thereby suppressing neuroblastoma development. This tumor-suppressive role is reflected by the fact that DPP-4 expression is greatly decreased and often lost in neuroblastoma cells as compared to readily detectable in vivo expression of DPP-4 in normal neural crest-derived structures from which neuroblastoma is believed to originate. Furthermore, experimental restoration of DPP-4 expression in neuroblastoma cells led to their differentiation, decreased migration and lower angiogenic potential, as well as to apoptosis (Arscott et al., 2008).

[0033] The above facts strongly indicate that a careful consideration should be taken before attempting to re-purpose the currently approved for diabetic patients DPP-4-inhibitors for treating neurological cancers, including GBMs and neuroblastomas, as there exists an increased probability of triggering opposite than the intended outcomes that may be ultimately detrimental to the patient if their cancer genetics is not appropriate for such treatments. Hence, there exist a need for reliable responsive-patient identification method and for accelerated development and approval of tailor-made gene target specific therapies for neurological cancer patients, optimally with already approved drugs with verified safety profiles.

[0034] MULTIPLE MYELOMA DISEASE

[0035] Multiple myeloma (MM) is a form of cancer characterized by the uncontrolled proliferation of plasma cells in blood-forming tissue, namely the bone marrow. It is arising from proliferation of monoclonal plasma cells from B cell populations in germinal centres which migrate to the bone marrow, wherein the plasma cells are the white blood cells, which plays critical role in the immune system by producing antibodies to fight infections. In MM, these plasma cells become cancerous and accumulate in the bone marrow, disrupting the production of normal blood cells.

[0036] The pathogenesis of multiple myeloma involves genetic, epigenetic, and microenvironmental factors. Genetic alterations such as chromosomal translocations, deletions, and mutations play a key role in disease initiation and progression. Epigenetic modifications, including changes in DNA methylation and histone acetylation, further contribute to the dysregulation of gene expression. The bone marrow microenvironment also plays a critical role by providing a supportive niche for myeloma cell survival and proliferation. Interactions between myeloma cells and the bone marrow stroma activate various signaling pathways, such as the NF-KB pathway, PI3K / Akt / mTOR signaling, and JAK / STAT signaling, which induces tumor growth and drug resistance.

[0037] MM-affected patients can experience a variety of disease-related symptoms due to, bone marrow infiltration, bone destruction, renal failure, immunodeficiency, and the psychosocial burden of a cancer diagnosis. This abnormal proliferation leads to a variety of clinical manifestations, including anemia, bone lesions, hypercalcemia, renal impairment, and an increased risk of infections.MM is the second most common blood cancer and represents a significant cause of morbidity and mortality worldwide. MM places significant burdens on the healthcare system and the society in general. For examples, MM accounts for up to 10% of all hematological malignancies. The estimated prevalence of MM in the US was -126% increase in cases from 1996 to 2016, while in 2020 it was 32,270 new cases and 12,830 deaths due to MM.

[0038] The current standard of care includes a combination of proteasome inhibitors, immunomodulatory drugs, monoclonal antibodies, corticosteroids, and autologous stem cell transplantation. Recent advancements in molecular biology and immunology have led to the identification of novel therapeutic targets. For instance, the development of monoclonal antibodies targeting surface antigens such as CD38 and BCMA, and additional therapies aimed at modulating the immune system, such as chimeric antigen receptor (CAR) T-cell therapy and bispecific T-cell engagers (BiTEs). Despite these advancements, challenges such as immune escape mechanisms, off-target effects and limited efficacy in certain patient populations persist, which ultimately leads the disease to often relapses and becomes refractory to the treatments.

[0039] Hence, there remains an unmet need for an innovative therapy which addresses the challenges and can provide improved efficacy, reduced toxicity, and has the potential to overcome resistance mechanisms in the treatment of multiple myeloma.

[0040] SUMMARY

[0041] The present invention relates to gliptins, the inhibitors of dipeptidyl peptidase 4, also known as DPP-4 inhibitors, for use in the treatment and / or prevention of a MALT-1 mediated or involving disease in a subject, preferably selected from allergic inflammatory disease, an autoimmune disease, preferably multiple sclerosis, multiple myeloma and glioma, in a human subject. Preferably, the disease is any one or more of allergic inflammatory disease, multiple sclerosis, multiple myeloma and glioma. The present invention relates to a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating a MALT-1 mediated disease in a subject or a MALT-1 involving disease in a subject, preferably selected from allergic inflammatory disease, an autoimmune disease, preferably multiple sclerosis, multiple myeloma and glioma, in a subject, preferably a human subject in need thereof.

[0042] Reference will now be made in detail to general aspects and specific embodiments of the disclosed herein concepts, which unless specified otherwise, are to be assumed to be interrelated with each other and suitable to work in co-operation. While specific embodiments will be described, it will be understood that they are not intended to be read in isolation from other embodiments. On the contrary, the presented herein disclosure is intended to address and / or cover all alternatives, modifications, and equivalents, which may be included within the scope of the appended claims. One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice and should be construed as falling within the scope of the appended claims.The general concept underlying the present disclosure originates from an unexpected finding of the inventors that gliptins, which are known to be DPP-4 inhibitors, can bind to an allosteric site on MALT1 and inhibit the same, advantageously reversibly.

[0043] Furthermore, but without wishing to be bound to the theory, it surprisingly appears that although gliptins were developed for targeting a disease target located extracellularly (catalytic domain of DPP-4) they seem to be inhibiting and allosterically binding to MALT1 (at least partially) intracellularly.

[0044] These surprising and unexpected findings lead the inventors to the presented herein novel use of the gliptins for the treatment of MALT1 -positive, MALT-1 -mediated diseases such as allergic inflammatory disease, cancers such as neurological cancers, multiple myeloma, autoimmune diseases such as MS.

[0045] A possible advantage of the presented herein approaches is that the currently known canonical MALT1 inhibitors display certain immunity-related side effects, which are either not known for one or more of gliptins, in particular the already approved gliptins. More details about MALT1 and its inhibition is explained below.

[0046] MALT-1 MEDIATED DISEASE

[0047] An aspect of the disclosure relates to a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating a MALT-1 mediated disease selected from:

[0048] allergic inflammatory disease,

[0049] an autoimmune disease, preferably multiple sclerosis,

[0050] multiple myeloma, and

[0051] a neurological cancer, preferably glioma,

[0052] in a subject, preferably a human subject, in need thereof, or

[0053] for use in a therapeutic method of treating neurological cancer in a human subject in need of the treatment, wherein the neurological cancer is positive for expression of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1).

[0054] An embodiment is the pharmaceutical composition for use, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin or cetagliptin, preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, more preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin or alogliptin, most preferably wherein the gliptin is sitagliptin.An embodiment is the pharmaceutical composition for use, wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, or cetagliptin, preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin or alogliptin.

[0055] An embodiment is the pharmaceutical composition for use, wherein the gliptin is not sitagliptin. An embodiment is the pharmaceutical composition for use, wherein the gliptin is an inhibitor of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1), preferably an allosteric inhibitor of MALT1.

[0056] An embodiment is the pharmaceutical composition for use, wherein the therapeutic effect of the gliptin comprises and / or results from inhibition of MALT1 , preferably wherein the inhibition of MALT1 is an allosteric inhibition resulting from binding of the gliptin to an allosteric site within MALT1.

[0057] An embodiment is the pharmaceutical composition for use, wherein the neurological cancer is negative for expression of dipeptidyl peptidase-4 (DPP-4) or comprises DPP-4 expression considered as unchanged to or lower as compared to a healthy reference tissue, preferably being the tissue from which the cancer originated and / or non-cancerous tissue surrounding the cells of the cancer, advantageously being a healthy tissue of the subject.

[0058] An embodiment is the pharmaceutical composition for use, wherein MALT1 expression in the neurological cancer is assessed as MALT1 overexpression.

[0059] An embodiment is the pharmaceutical composition for use, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin; preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin; more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin, and alogliptin, most preferably wherein the gliptin is sitagliptin.

[0060] An embodiment is the pharmaceutical composition for use, wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, and cetagliptin; preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin; more preferably wherein the gliptin is of vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin; even more preferably of linagliptin, vildagliptin, saxagliptin and alogliptin, most preferably wherein the gliptin is linagliptin.

[0061] An embodiment is the pharmaceutical composition for use, wherein the method comprises oral administration of the pharmaceutical composition.

[0062] An embodiment is the pharmaceutical composition for use, wherein the method results in inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside aberrant cells, preferably aberrant Th2 cells or preferably aberrant Th1 cells and / or Th17 cells, or preferably inhibition of MALT1 activity inside myeloma cells, or inhibition of MALT1 -mediated activation of NF-KB signalingpathway, or inhibition of cytokines regulated by MALT1 -mediated NF-KB signaling, such as TNF-a and / or IL-6.

[0063] An embodiment is the pharmaceutical composition for use, wherein:

[0064] the allergic inflammatory disease is a chronic allergic condition and / or a late-phase allergic condition;

[0065] the MS is clinically isolated syndrome (CIS), relapsing-remitting MS (RRMS), secondary progressive MS (SPMS), primary progressive MS (PPMS) or progressive relapsing MS (PRMS).

[0066] An embodiment is the pharmaceutical composition for use, wherein:

[0067] the allergic inflammatory disease is selected from the group consisting of allergic asthma, anaphylaxis, sinusitis, allergic rhinitis, eczema, hives, food allergy, chronic urticaria, atopic dermatitis and ocular allergic diseases;

[0068] the MS is at pre-symptomatic stage, clinically isolated syndrome stage, relapsing-remitting stage, or secondary progressive stage.

[0069] An embodiment is the pharmaceutical composition for use, wherein the MALT-1 mediated disease selected from allergic inflammatory disease, an autoimmune disease, preferably multiple sclerosis, multiple myeloma and a neurological cancer such as glioma is mediated by or dependent on MALT1 activity.

[0070] An embodiment is the pharmaceutical composition for use, wherein the method comprises the step of diagnosing a subject as suffering from the MALT-1 mediated disease selected from allergic inflammatory disease, an autoimmune disease, preferably multiple sclerosis, multiple myeloma and a neurological cancer such as glioma, or being at risk of suffering from the MALT-1 mediated disease selected from allergic inflammatory disease, an autoimmune disease, preferably multiple sclerosis, multiple myeloma and a neurological cancer such as glioma, mediated by or dependent on MALT1 activity.

[0071] An embodiment is the pharmaceutical composition for use, wherein the neurological cancer is brain cancer and / or is glioma, neuroblastoma, or medulloblastoma; preferably being glioma or neuroblastoma, more preferably being glioblastoma or gliosarcoma, even more preferably being malignant glioblastoma, most preferably being glioblastoma multiforme.

[0072] An embodiment is the pharmaceutical composition for use, wherein the treatment comprises administering to the human subject of an effective amount of the compound, preferably directly into the neurological cancer or into the cavity remaining after surgical removal of the neurological cancer, possibly in combination with one or more chemotherapeutic agent and / or anticancer agent specific to the cancer of the subject.

[0073] An embodiment is the pharmaceutical composition for use, wherein the treatment is followed by radiotherapy and / or chemotherapy.

[0074] An embodiment is the pharmaceutical composition for use, wherein MALT1 expression status, and optionally DPP-4 expression status, comprises RNA expression analysis and / or protein expressionanalysis, preferably by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and / or immunoassay.

[0075] An embodiment is the pharmaceutical composition for use, wherein the method comprises the oral administration of the gliptin, pharmaceutically acceptable salt thereof or hydrate thereof, at a dose of 0.01 mg to 1000 mg per day.

[0076] An embodiment is the pharmaceutical composition for use, wherein the pharmaceutical composition is provided in an oral unit dosage form, preferably in the form of a tablet, comprising the gliptin, the pharmaceutically acceptable salt thereof or hydrate thereof, in an amount of 0.01 mg to 1000 mg.

[0077] An aspect of the disclosure relates to a therapeutic or prophylactic method of treating a MALT-1 mediated disease selected from allergic inflammatory disease, multiple sclerosis, multiple myeloma and glioma, in a subject, preferably human subject, in need thereof, said method comprising the step of administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0078] An aspect of the disclosure relates to a use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of a MALT-1 mediated disease selected from allergic inflammatory disease, multiple sclerosis, multiple myeloma and glioma, in a subject, preferably human subject, in need thereof.

[0079] An aspect of the disclosure relates to an in vitro method for determining an increased probability that a neurological cancer in a human subject will respond to the treatment with

[0080] a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, wherein the gliptin is selected from wherein the gliptin is selected from any one or more of sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, , cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin, and cetagliptin; the method comprising the steps of:

[0081] - providing a test first sample obtained from the subject;

[0082] - detecting the presence of MALT1 expression and preferably assessing the expression level of MALT 1 in the first test sample;

[0083] - preferably, comparing the assessed expression level of MALT1 to a reference MALT1 expression level;

[0084] wherein finding that MALT1 expression is present in the first test sample and preferably that the expression level of MALT1 assessed in the first test sample is higher as compared to the reference MALT 1 expression level is indicative of an increased probability that the subject will respond to the treatment.

[0085] An embodiment is the in vitro method, further comprising the steps of:

[0086] - providing a second test sample obtained from the subject;

[0087] - detecting the presence of DPP-4 expression or optionally assessing the expression level of DPP-4 or DPP-4 enzymatic activity level in the second test sample;- preferably, comparing the assessed expression level or enzymatic activity level of DPP-4 to a reference DPP-4 expression level or a reference DPP-4 enzymatic activity level, respectively; wherein finding that DPP-4 expression is absent in the second test sample or optionally that the expression level of DPP-4 or DPP-4 enzymatic activity level assessed in the second test sample is lower as compared to the reference DPP-4 expression level or a reference DPP-4 enzymatic activity level, respectively; preferably finding that DPP-4 expression is absent in the second test sample is indicative of an increased probability that the subject will respond to the treatment, wherein the response is further associated with an increased probability of not causing adverse DPP-4-inhibition-associated side effects.

[0088] An embodiment is the in vitro method, wherein the first test sample and / or the second test sample is or comprises biopsy sample, cerebrospinal fluid (CSF) sample, or a blood sample, preferably being biopsy sample or the cerebrospinal fluid (CSF) sample, even more preferably being biopsy sample.

[0089] An embodiment is the in vitro method, wherein detecting the presence of expression and / or assessing the expression level comprises RNA analysis, preferably comprising RNA-sequencing or reverse transcription-quantitative polymerase chain reaction (RT-qPCR), more preferably comprising RT-qPCR.

[0090] An embodiment is the in vitro method, wherein detecting the presence of expression and / or assessing the expression level comprises protein analysis, preferably comprising an immunoassay, more preferably comprising ELISA.

[0091] An embodiment is the in vitro method, wherein the first test sample and the second test sample are the same test sample, preferably wherein detecting the presence of MALT1 and DPP-4 expression, optionally assessing the expression level of MALT1 and DPP-4 is performed as part of the same assay, more preferably being a multiplex RT-qPCR assay or an immunoassay.

[0092] An embodiment is the in vitro method, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin;

[0093] preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin;

[0094] more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin, and alogliptin, most preferably wherein the gliptin is sitagliptin;

[0095] and / or

[0096] wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, and cetagliptin;

[0097] preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin;

[0098] more preferably wherein the gliptin is of vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin;even more preferably of linagliptin, vildagliptin, saxagliptin and alogliptin,

[0099] most preferably wherein the gliptin is linagliptin.

[0100] An embodiment is the in vitro method, wherein the reference MALT1 expression value, and optionally the reference DPP-4 expression value, is defined and has (have) been predetermined based on assessing MALT1 expression levels, and optionally DPP-4 expression levels, respectively, in a plurality of control samples obtained from healthy control subjects.

[0101] An embodiment is the in vitro method, wherein the reference MALT1 expression value, and optionally the reference DPP-4 expression value, is determined based on assessing normal MALT1 expression levels, and optionally normal DPP-4 expression levels, respectively, in a healthy tissue sample obtained from the subject, preferably being a healthy tissue corresponding to the tissue from which the cancer originated, optionally being healthy non-cancerous tissue surrounding the cancer cells.

[0102] An embodiment is the in vitro method, wherein the expression level of MALT1 assessed for the first test sample is at least 2 fold higher than the reference MALT1 expression value, preferably being at least 5 fold higher; more preferably being at least 10 fold higher.

[0103] ALLERGIC INFLAMMATORY DISEASE

[0104] The invention relates to a pharmaceutical composition for use in the treatment of allergic inflammatory disease.

[0105] Our immune system plays an important role in sensing of environmental conditions to protect us against pathogens including microorganisms, foreign substances or defect cells of our own body. It is a tightly regulated network of different cell types and disorders of the immune system can result in lymphoma development or serious autoimmune diseases like multiple sclerosis. The human immune system is divided into the innate and adaptive response. The NF-KB pathway has an important regulatory function in both systems and is involved in their communication. The CBM-complex is one central regulatory assembly in the NF-KB signalling cascade and consists of CARMA1 , BCL-10 and MALT1. Mutations in one of these genes are connected with the development of lymphoma and autoimmune diseases (Hailfinger, 2009; Me Guire, 2013). MALT1 harbors a paracaspase domain, whose proteolytic activity is involved in NF-KB signal transduction (Schlauderer; 2015).

[0106] NF-KB has been known for more than 30 years. NF-KB (nuclear factor) is a protein factor with gene transcriptional regulation, which is present in almost all nucleated cells. When cells are stimulated by inflammatory mediators, NF-KB protein is activated in the cytoplasm and enters the nucleus to regulate the expression of various inflammatory factors, playing an important role in allergic diseases. For example, in allergic asthma, the NF-KB / MAPK pathway controls inflammatory and immune responses by regulating TNF-a and IL-6. A 2019 study demonstrated that after nuclear translocation of phosphorylated P65, inhibited NF-KB / MAPK signalling may modulate IgE and IL-4 production. Enhanced NF-KB nuclear binding or production was also found in inflammatory cells collected from induced sputum in asthmatic patients. Besides, experiments have shown enhanced NF-KB activation in airway tissues and inflammatory cells challenged by allergens such as ovalbumin (OVA) and house dustmite (HDM) extract. In addition, the studies related to increased Tfh cells in allergic diseases, NF-KB deficiency led to a decrease in CXCR5 (Tfh cells expressing chemokine receptors) in mice and a consequent decrease in the number of Tfh cells. All these studies suggest that NF-KB signaling is essential in the cellular immune response of allergic diseases (Wang 2023).

[0107] The inventors now surprisingly established that gliptins are inhibitors of intracellular MALT1 , opening up a whole new avenue for developing new treatments for allergic inflammatory disorders. Interfering in the NF-KB pathway by inhibiting MALT1 has been established by the inventors, resulting in silencing or killing of the immune cells (Th2) involved in the e.g. chronic allergic condition and / or a late-phase allergic condition of allergic inflammatory disorders.

[0108] A first aspect if the invention relates to a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating allergic inflammatory disease in a subject, preferably human subject in need thereof.

[0109] A second aspect of the invention relates to a therapeutic or prophylactic method of treating allergic inflammatory disease in a subject, preferably a human subject in need thereof, said method comprising the administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0110] A third aspect of the invention relates to the use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of allergic inflammatory disease in a subject, preferably a human subject in need thereof.

[0111] An embodiment is the pharmaceutical composition for use according to the invention, wherein the gliptin is Sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin, Alogliptin, Teneligliptin, Trelagliptin, Omarigliptin, Evogliptin, Gosogliptin, Retagliptin, Denagliptin, Cofrogliptin, Fotagliptin, Prusogliptin, Dutogliptin, Neogliptin or Cetagliptin, preferably wherein the gliptin is Sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin, Alogliptin, Teneligliptin, Trelagliptin, Omarigliptin, Evogliptin, Gosogliptin, Retagliptin, Denagliptin, Cofrogliptin, Fotagliptin, Prusogliptin, more preferably wherein the gliptin is sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin or Alogliptin, more preferably wherein the gliptin is Sitagliptin, Linagliptin, Vildagliptin, Saxagliptin or Alogliptin, most preferably wherein the gliptin is sitagliptin.

[0112] An embodiment is the pharmaceutical composition for use according to the invention, wherein the gliptin is Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin, Alogliptin, Teneligliptin, Trelagliptin, Omarigliptin, Evogliptin, Gosogliptin, Retagliptin, Dutogliptin, Neogliptin, Denagliptin, Cofrogliptin, Fotagliptin, Prusogliptin, or Cetagliptin, preferably wherein the gliptin is Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin or Alogliptin, more preferably wherein the gliptin is Vildagliptin, Saxagliptin, Linagliptin or Alogliptin.

[0113] An embodiment is the pharmaceutical composition for use according to the invention, wherein the gliptin is not Sitagliptin.

[0114] An embodiment is the pharmaceutical composition for use according to the invention, wherein the gliptin is an inhibitor of MALT1 , preferably an allosteric inhibitor of MALT1.An embodiment is the pharmaceutical composition for use according to the invention, wherein the method comprises oral administration of the pharmaceutical composition.

[0115] An embodiment is the pharmaceutical composition for use according to the invention, wherein the method results in inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside aberrant cells, preferably aberrant Th2 cells.

[0116] An embodiment is the pharmaceutical composition for use according to the invention, wherein the allergic inflammatory disease is a chronic allergic condition and / or a late-phase allergic condition.

[0117] An embodiment is the pharmaceutical composition for use according to the invention, wherein the allergic inflammatory disease is selected from the group consisting of allergic asthma, anaphylaxis, sinusitis, allergic rhinitis, eczema, hives, food allergy, chronic urticaria, atopic dermatitis and ocular allergic diseases.

[0118] An embodiment is the pharmaceutical composition for use according to the invention, wherein the allergic inflammatory disease is mediated by or dependent on MALT1 activity.

[0119] An embodiment is the pharmaceutical composition for use according to the invention, wherein the method comprises the step of diagnosing a subject as suffering from or being at risk of suffering from allergic inflammatory disease mediated by or dependent on MALT1 activity.

[0120] AUTOIMMUNE DISEASE, PREFERABLY MS

[0121] The present invention resides in the unexpected finding of the inventors that gliptins bind MALT1 and in particular, bind MALT1 allosterically and inhibit the same.

[0122] In an aspect, the invention provides a therapeutic or prophylactic method of treating MS in a human subject in need thereof, said method comprising the administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0123] In another aspect, the invention provides a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating multiple sclerosis (MS) in a human subject in need thereof.

[0124] In yet a further aspect, the invention provides the use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of MS in a human subject in need thereof. In a further aspect, the present disclosure relates to a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof that is suitable and / or specifically adapted for uses in accordance with the invention.

[0125] In specific embodiments of the invention, the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin or cetagliptin, preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, more preferably the gliptin is sitagliptin, vildagliptin, saxagliptin,linagliptin, gemigliptin, anagliptin or alogliptin, more preferably the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin or alogliptin, most preferably the gliptin is sitagliptin.

[0126] In further embodiments of the invention, the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, or cetagliptin, preferably the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably the gliptin is vildagliptin, saxagliptin, linagliptin or alogliptin.

[0127] In an embodiment, the gliptin is not sitagliptin.

[0128] In an embodiment, the gliptin is an inhibitor of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1), preferably an allosteric inhibitor of MALT1.

[0129] In a further embodiment, the method comprises oral administration of the pharmaceutical composition.

[0130] In an embodiment, the method results in inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside aberrant cells, preferably aberrant Th1 cells, Th 17 cells.

[0131] In another embodiment, the MS is clinically isolated syndrome (CIS), relapsing-remitting MS (RRMS), secondary progressive MS (SPMS), primary progressive MS (PPMS) or progressive relapsing MS (PRMS).

[0132] In a further embodiment, the MS is at pre-symptomatic, clinically isolated syndrome, relapsingremitting, or secondary progressive stage.

[0133] In an embodiment, the MS is mediated by or dependent on MALT1 activity.

[0134] In yet another embodiment, the method comprises the step of diagnosing a subject as suffering from or being at risk of suffering from MS mediated by or dependent on MALT1 activity.

[0135] In an embodiment, the method comprises the oral administration of the gliptin, pharmaceutically acceptable salt thereof or hydrate thereof, at a dose of 0.01 mg to 1000 mg per day.

[0136] In an embodiment, the pharmaceutical composition is provided in an oral unit dosage form, preferably in the form of a tablet, comprising the gliptin, the pharmaceutically acceptable salt thereof or hydrate thereof, in an amount of 0.01 mg to 1000 mg.

[0137] NEUROLOGICAL CANCER, PREFERABLY GLIOMA

[0138] The above-identified need is herewith addressed for a specific subpopulation of neurological cancer patients and is based on an unexpected finding of the inventors that sitagliptin, together with several other gliptins, are capable of allosteric binding and efficient inhibition of mucosa-associated lymphoid tissue lymphoma translocation protein 1 , known as MALT1.

[0139] MALT1 is an intracellular matrix-associated paracaspase previously linked to antigen receptor-mediated NF-KB activation and B-cell lymphoma. In addition to its scaffold-related role in NF-KB activation, MALT1 regulates cell adhesion, mRNA stability, and mTOR signaling through its proteolytic activity (Rebeaud et al, 2008; Staal et al, 2011 ; Uehata et al, 2013; Hamilton et al, 2014; Jeltsch et al, 2014; Nakaya et al, 2014). MALT1 has been shown to be constitutively active in activated B-cell-likediffuse large B-cell lymphoma (ABC DLBCL), with its inhibition being lethal (Ngo et al, 2006; Hailfinger et al, 2009; Nagel et al, 2012).

[0140] Like DPP-4, MALT1 is a druggable molecular target with several known specific inhibitors. These include the early / first-generation irreversible catalytic inhibitors such as z-VRPR-fmk and z-LVSR-fmk, and the newer-generation allosteric (noncompetitive) reversible MALT1 inhibitors including highly-MALT1 specific phenothiazine derivatives, such as mepazine, thioridazine, and promazine (Jaworski & Thome, 2015). As under physiolgical conditions MALT1 activity is required not only for the immune response, but also for the development of natural T reg cells that keep the immune response in check, unfortunately, most of these inhibitors are associated with more or less severe immune deregulation side effects, such as immunodeficiency or, at the opposite end of the spectrum, autoinflammation (Gewies et al. 2014).

[0141] Unlike DPP-4, however, whose role in neurological cancer progression is at most questionable, leaning towards a more tumor-suppressive role, MALT 1 is an oncogene with an established involvement in glioblastoma. Pharmacological inhibition of in vitro and in vivo MALT1 activity in glioblastoma cells was demonstrated to increase the amounts of endo-lysosomes, impair autophagic flux, and culminate in lysosomal-mediated cell death, concomitantly with mTOR inactivation and dispersion from endo-lysosomes (Jacobs et al., 2019). Next, MALT1 inhibitor treatment was shown to result in increased immuno-reactivity of GBM-associated macrophages and reduced GBM tumor growth (Azambuja et al., 2024).

[0142] In view of the above, the inventors’ surprising discovery that gliptins are allosteric inhibitors of MALT 1 has now opened up a whole new avenue for developing new treatments for neurological cancers characterized by MALT1 (over)expression, and advantageously also by base-line low or lost DPP-4 expression, which treatments, given the current knowledge of the approved gliptin-safety profiles, will advantageously be devoid of the most severe side effects associated with the previously known MALT1 inhibitors.

[0143] In line with the above, in a first general aspect, provided herein is a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic method of treating neurological cancer (preferably brain cancer) in a human subject in need of the treatment, wherein the cancer is positive for expression of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1).

[0144] In another aspect, the invention provides a therapeutic or prophylactic method of treating neurological cancer (preferably brain cancer) in a human subject in need of the treatment, said method comprising the administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, wherein the cancer is positive for expression of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1).

[0145] In yet a further aspect, the invention provides the use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of neurological cancer (preferably braincancer) in a human subject in need of the treatment, wherein the cancer is positive for expression of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1).

[0146] In some exemplary advantageous embodiments, the neurological cancer overexpresses MALT1 and / or is negative for expression of dipeptidyl peptidase-4 (DPP-4) or comprises DPP-4 expression considered as unchanged to or lower as compared to a healthy reference tissue.

[0147] In a second general aspect related to the above one, an in vitro method for determining an increased probability that a neurological cancer (preferably brain cancer) in a human subject will respond to the treatment with the pharmaceutical composition of the present disclosure the method comprising the steps of:

[0148] - providing a test first sample obtained from the subject;

[0149] - detecting the presence of MALT1 expression and preferably assessing the expression level of MALT 1 in the first test sample;

[0150] - preferably, comparing the assessed expression level of MALT1 to a reference MALT1 expression level;

[0151] wherein finding that MALT1 expression is present in the first test sample and preferably that the expression level of MALT1 assessed in the first test sample is higher as compared to the reference MALT1 expression level is indicative of an increased probability that the subject will respond to the treatment with the pharmaceutical composition of the present disclosure.

[0152] These aspects and advantageous embodiments thereof are described in more detail herein below.

[0153] MULTIPLE MYELOMA

[0154] The inventors surprisingly established that gliptins are inhibitors of intracellular MALT1 , opening up a whole new avenue for developing new treatments for multiple myeloma patients. Interfering in the NF-KB pathway by inhibiting MALT1 has been established by the inventors, resulting in silencing or killing of the tumor cells involved in multiple myeloma.

[0155] In one aspect, the present disclosure relates to a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating multiple myeloma disease in a human subject in need thereof.

[0156] In another aspect, the present disclosure relates to a therapeutic or prophylactic method of treating multiple myeloma disease in a human subject in need thereof, said method comprising the administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0157] In yet another aspect, the present disclosure relates to use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of multiple myeloma disease in a human subject in need thereof.In one embodiment, the multiple myeloma disease is mediated by or dependent on MALT1 activity.

[0158] In one embodiment, the gliptin is an inhibitor of MALT1 , preferably an allosteric inhibitor of MALT1.

[0159] In another embodiment, the gliptins are selected from one or more of: sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin or cetagliptin, preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, more preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin or alogliptin, most preferably wherein the gliptin is sitagliptin.

[0160] DEFINITIONS

[0161] A “MALT1 inhibitor1’ has its regular scientific meaning and here refers to a compound which inhibits and / or downregulates the activity of MALT1 , Mucosa-associated lymphoid tissue lymphoma translocation protein 1.

[0162] An “allosteric MALT1 inhibitor1’ has its regular scientific meaning and here refers to a compound which binds to the allosteric site of MALT1 . Said allosteric site is situated in the interface between the paracaspase and Ig3 domain connecting helix a1lg3 of MALT1 , that is far from the active site of the enzyme. The allosteric inhibitor which can bind to said allosteric site of the protein are noncompetitive and reversible inhibitors. The inhibition can be effective on the scaffold function or protease function or both. A suitable method for determining the ability of a compound to bind to the allosteric site in suit is described in example 1 . A negative control for determining allosteric binding of gliptins to MALT 1 , can suitably be provided by a compound of formula VI:

[0163]

[0164] Formula VI;

[0165] the same is represented as molecule 11 in Tran, T.D, et al. (2019).

[0166] For completeness, established (competitive) MALT 1 inhibitors, such as mepazine / S-mepazine, MI-2 or MLT-748 are shown below.Mepazine and S-mepazine as referred to herein may be represented by compounds of formula VII and formula VII’, respectively:

[0167]

[0168] Formula VII Formula VII’

[0169] MI-2 as referred herein may be represented by compound of formula VIII:

[0170]

[0171] Formula VIII

[0172] MLT-748 as referred herein may be represented by compound of formula IX:

[0173]

[0174] A “MALT1 mediated disease" or “MALT-1 involving disease" or “multiple allergic inflammatory disease mediated by MALT1” has its regular scientific meaning and here refers to a disease mediated by or dependent on MALT 1.

[0175] A “Th17-mediated disease” has its regular scientific meaning and here refers to a disease mediated by or dependent onT-helper17 (Th17) cell activity.

[0176] The term “gliptin” or “gliptins” as used herein refers to a class of pharmaceutical compounds which are the inhibitors of dipeptidyl peptidase 4, also known as DPP-4 inhibitors, and includes any given gliptin in any form, including, in particular, all pharmaceutically acceptable salt forms, as well as hydrates / solvates thereof. Primarily the gliptins function by inhibiting the activity of the DPP-4 enzyme, which plays a role in glucose metabolism, hence the gliptins are widely used as hypoglycemic agents, primarily in the management of type 2 diabetes mellitus. Several gliptins have been developed and even approved by various drug regulatory bodies across the globe. An overview for different types of gliptins, their formulas and synthesis pathways can be found in Kushwaha, Ram & Haq, Wahajul & Katti, Seturam. (2014), which is herewith incorporated by reference. For the purpose of present disclosure, a non-exhaustive list of gliptins includes: sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, gosogliptin, alogliptin, trelagliptin, carmegliptin, omarigliptin, evogliptin, dutogliptin, neogliptin, retagliptin, melogliptin, denagliptin, cofrogliptin, fotagliptin or prusogliptin, cetagliptin.

[0177] Sitagliptin as referred herein may also be represented by compound of formula I:

[0178]

[0179] Formula I

[0180] Alogliptin as referred herein may also be represented by compound of formula II:

[0181]

[0182] Formula II

[0183] Linagliptin as referred herein may also be represented by compound of formula III:

[0184]

[0185] Vildagliptin as referred herein may also be represented by compound of formula IV:

[0186]

[0187] Formula IV

[0188] Saxagliptin as referred herein may also be represented by compound of formula V:

[0189]

[0190] As used herein, the term “cancer” has its regular meaning from the field of medicine and can be construed as referring to any one of diverse diseases belonging to a large family of pathologiescharacterized by the uncontrolled growth of cells in a body. Representative forms of cancer include but are not limited to carcinomas, sarcomas, myelomas, leukemias, lymphomas, etc.

[0191] Within particularly preferred embodiments of the present disclosure, the cancer is a neurological cancer, advantageously is a brain cancer (which is a term that can be seen as synonymous to the terms “brain tumor” and “intracranial neoplasm”). As used herein, the term “neurological cancer” has its regular meaning as it would be understood by a medical practitioner practicing in the field of oncology and can be construed as referring to a cancerthat originated from a cell of the brain or spine or from any nerve cell, even at a very early stage. Most neurological cancers at the moment of their detection will still be localized within the central nervous system (or as sometimes used herein, the “CNS”), usually in the brain (including the cerebellum and the brain stem in addition to the brain proper), with rare exceptions such as neuroblastoma, being a malignancy of neural crest cells that give rise e.g. to the sympathetic nervous system, which fact determines that neuroblastomas may occur at any site within the sympathetic nervous system, most commonly in the abdomen. As it will be understood by the medical practitioner practicing in the field of oncology, depending on the stage at which a neurological cancer was detected, it may have metastasized to another part of the body. Hence, as used herein, the term neurological cancer is to be construed as also encompassing metastases of neurological cancers localized outside their place of origin (e.g. the CNS or the sympathetic nervous system). Especially that, according to certain embodiments of the present disclosure, such metastases may constitute attractive neurological cancers for being treated with the presented herein new therapeutic approaches for the reason that in their selection for genetic changes that lead to their increased invasiveness, they are more likely to have acquired overexpression of MALT1 and / or silencing (or otherwise losing) the expression of DPP-4 and therefore are possibly associated with an increased probability of responding to a treatment with the presented herein pharmaceutical compositions.

[0192] As used herein, the term “inhibitor1’ has its regular scientific meaning and can be construed as referring to a molecule like a compound which blocks (inhibits) and / or interferes with (e.g. slows down) activity of another, usually larger molecule, such as an enzyme or a receptor. For example, as used herein “MALT1 inhibitor” is to be construed as an inhibitor of mucosa-associated lymphoid tissue lymphoma translocation protein 1.

[0193] As used herein, the term “approximately” or “about,” as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In certain embodiments, the term “approximately” or “about” refers to a range of values that fall within 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

[0194] The term “comprising”, used in the claims, should not be interpreted as being restricted to for example the elements or the method steps or the constituents of a compositions listed thereafter; it does not exclude other elements or method steps or constituents in a certain composition. It needs to be interpreted as specifying the presence of the stated features, integers, (method) steps or components as referred to, but does not preclude the presence or addition of one or more other features, integers, steps or components, or groups thereof. Thus, the scope of the expression “a method comprising stepsA and B” should not be limited to a method consisting only of steps A and B, rather with respect to the present disclosure, the only enumerated steps of the method are A and B, and further the claim should be interpreted as including equivalents of those method steps. Thus, the scope of the expression “a composition comprising components A and B” should not be limited to a composition consisting only of components A and B, rather with respect to the present disclosure, the only enumerated components of the composition are A and B, and further the claim should be interpreted as including equivalents of those components.

[0195] In addition, reference to an element or a component by the indefinite article "a" or "an" does not exclude the possibility that more than one of the element or component are present, unless the context clearly requires that there is one and only one of the elements or components. The indefinite article "a" or "an" thus usually means "at least one".

[0196] The use of terms in brackets in the text, with the exception of chemical and / or mathematical formulae, usually means that the term within brackets specifies a possible option or a possible meaning and should thus not be considered limiting.

[0197] The embodiments as described herein can operate in combination and cooperation, unless specified otherwise. Furthermore, the various embodiments, although referred to as “preferred” or “e.g.” or “for example” or “in particular” and the like are to be construed as exemplary manners in which the disclosed herein concepts may be implemented rather than as limiting.

[0198] For all Figures, “Figure” and “Fig.” refer to the same.

[0199] The term “subject” or “individual” as used herein refers to a human suffering from or at risk of a certain health-related disorder, such as a disease or other pathological condition. The term “subject” and “individual” herein are used interchangeably. The terms “subject”, “human subject”, “individual”, and “patient” as used herein are synonymous and can be used interchangeably and are to be construed as referring to a person suffering from or at risk of a certain health-related disorder, such as a disease or other pathological condition.

[0200] The term “treatment” as used herein has its conventional meaning and refers to a medical intervention or management of a subject with the intention to cure, ameliorate, stabilize, or prevent a health-related disorder. This term “treatment” includes e.g. active treatment that is a type of an action directed specifically toward the improvement of a health-related disorder, and also includes causal treatment that is a treatment directed towards a removal of the cause of the associated therewith health-related disorder.

[0201] The term ‘prophylaxis’ as used herein means a medical intervention or management of a subject with the intention to maintain health or the normal bodily functions. As used herein, prophylaxis is to be construed as falling within the scope of a treatment, unless indicated otherwise.

[0202] As used herein the term “administration” is to be construed as referring to the way of providing a substance, such as compound, or a pharmaceutical composition to a subject. Conversely, as used herein the term “delivery” is to be construed as referring to the way a compound reaches its destination site, e.g. specific zone, cell or tissue type like retina of the eye. For example, as used herein, administration can relate to intrathecal, intravenous, topical, intranasal, intraocular, etc. way of providing a compound into the subject’s body, with an intended delivery to e.g. cerebellum or the retina as thedestination site. Usually, the terms “administering” or “administration” will be construed as remating to the provision of a substance that is physiologically and / or (e.g., and) pharmacologically useful (e.g., to treat a condition in the subject).

[0203] The term “excipient” as used herein has its conventional meaning and refers to a pharmaceutically acceptable ingredient, which is commonly used in the pharmaceutical technology for preparing a granulate, solid or liquid oral dosage formulation.

[0204] The phrase “pharmaceutically acceptable” indicates that the substance or composition is compatible chemically and / or toxicologically, with the other ingredients comprising a formulation, and / or the subject being treated therewith.

[0205] The term “pharmaceutically acceptable salt” as used herein includes an acid addition salt or a base addition salt, formed with either organic or inorganic acids and / or base. Suitable pharmaceutically acceptable salts of the compounds of the invention include acid addition salts which may be salts of inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid and the like or of organic acids such as, for example, acetic acid, benzenesulfonic acid, methanesulfonic acid, benzoic acid, citric acid, lactic acid, fumaric acid, succinic acid, adipic acid, pimelic acid, suberic acid, azelaic acid, malic acid, tartaric acid, amino acids such as glutamic acid or aspartic acid, and the like. Suitable pharmaceutically acceptable salts of the compounds of the invention include base addition salts which may be salts of inorganic bases such as sodium, potassium, calcium, magnesium and the like, or of organic bases such as triethylamine, diethanolamine, ethanolamine, piperazine and the like. The pharmaceutically acceptable salts of the present invention include salts formed with the addition of one or more equivalents of acids and / or base, for example, monohydrochloride, dihydrochloride, mono sodium, disodium salts, and the like. As the skilled person would recognize, the list of possible inorganic acids, organic acids, inorganic bases, organic basis and amino acids is not exhaustive, but representative. Any further acids and / or basis and / or amino acids would fall under the purview of the present disclosure. Salts can be prepared by any process under the purview of an ordinary person skilled in the art (see Berge et al., J. Pharm. Sci. 1977, 66, 1 -19; and Handbook of Pharmaceutical Salts, Properties, and Use; Stahl and Wermuth, Ed.; Wiley-VCH and VHCA: Zurich, Switzerland, 2002).

[0206] The terms “multiple myeloma disease” and “multiple myeloma” have the same common meaning known in the art.

[0207] LEGEND TO THE FIGURES - BRIEF DESCRIPTION OF THE DRAWINGS

[0208] Figure 1A and B show the structure of the MALT1 allosteric site with in (A) MLT-748 bound thereto and with in (B) sitagliptin bound thereto.

[0209] Figure 2 shows the effects of S-mepazine and sitagliptin on Jurkat cell survival (allergic inflammation model).

[0210] Figure 3 shows the percentage of cell metabolism relative to untreated controls after treatment with (S)-mepazine (positive control) and sitagliptin at various concentrations (1 pM, 10 pM, and 50 pM).Figure 4 shows the effects of S-mepazine (5.0 pM) and sitagliptin (5.0 pM) on cytokine production, when Jurkat cells treated with MALT1 inhibitors were compared to untreated cells.

[0211] Figure 5 shows the antiproliferative effects of (S)-mepazine and sitagliptin in Jurkat cells mimicking Th1 / Th17 cell activation.

[0212] Figure 6 shows the percentage of cell metabolism relative to untreated controls after treatment with (S)-mepazine (positive control) and sitagliptin at various concentrations (1 pM, 10 pM, and 50 pM). Cell metabolism was assessed using the Alamar Blue assay. Error bars represent mean ± standard deviation from triplicate experiments.

[0213] Figure 7 shows the percentage of cytokine production including IL-17 and IFN-y when T cells are treated with (S)-mepazine (positive control) at the same concentration.

[0214] Figure 8: the effects of S-mepazine and sitagliptin on JJN3 cell survival.

[0215] Figure 9: dose dependent cell viability, proliferation and metabolism assay.

[0216] Figure 10: the effects of S-mepazine (5 pM) and sitagliptin (5 pM) on IL-6 and TNF-a cytokines production.

[0217] Figure 11 : antiproliferative effects of MI-2, mepazine and sitagliptin on U87-MG cell growth; Figure 12A: inhibition of MALT1 by sitagliptin at 10 pM in patient-derived glioblastoma cells (U3024);

[0218] Figure 12B: inhibition of MALT 1 by MI-2, mepazine and sitagliptin in patient-derived glioblastoma cells (U3024);

[0219] Figure 13A-C show the percentage of cell metabolism relative to untreated controls after treatment with (S)-mepazine (positive control) and sitagliptin at various concentrations (1 pM, 10 pM, and 50 pM). Cell metabolism was assessed using the Alamar Blue assay. Error bars represent mean ± standard deviation from triplicate experiments. Figure 13A: treatment of U3024 cells; Figure 13B: treatment of U3005 cells; and Figure 13C: treatment of U3028 cells.

[0220] DETAILED DESCRIPTION

[0221] The present invention is in the field of drug repurposing, wherein in an effort to address the unmet need for an improved treatment for a series of diseases such as MS, cancers such as multiple myeloma and glioma, and allergic inflammation, the present inventors have identified gliptins for use in the treatment of such diseases. In an unexpected finding, the inventors have now revealed that the gliptins, which are traditionally known to be DPP-4 inhibitors, can also bind to an allosteric site (pocket) on MALT1 and can act as an allosteric MALT1 inhibitor. This surprising and unexpected finding leads to a novel use of the gliptins for the treatment of diseases such as the aforementioned cancers, autoimmune disease and allergic inflammation.

[0222] The innovative concepts as presented herein will be described with respect to particular aspects and embodiments of the disclosure, which should be regarded as descriptive and not as limiting beyond of what is described in the claims. The aspects and / or the embodiments as described herein can operate in combination and cooperation, unless specified otherwise. While the disclosed herein innovativeconcepts are described with reference to these aspects and embodiments, it is contemplated that alternatives, modifications, permutations and equivalents thereof will become apparent to one having ordinary skill in the art upon reading the specification and upon contemplation of the drawings and / or graphs. The disclosed matter is not limited in any way to the illustrated embodiment and changes thereto can be made without departing from the scope which is defined by the appended claims.

[0223] The present invention relates to the unexpected finding of the inventors relating to gliptins, which are known to be DPP-4 inhibitors, and are now identified as having the capability to bind to an allosteric site on MALT1 and inhibit the enzymatic activity reversibly.

[0224] Moreover, although gliptins are developed for targeting a disease target located extracellularly (catalytic domain of DDP-4), the inventors now surprisingly found that these gliptins allosterically bind and inhibit MALT1 intracellularly.

[0225] ALLERGIC INFLAMMATION DISEASES

[0226] These surprising and unexpected findings lead to a novel use of the gliptins for the treatment of allergic inflammation diseases, particularly for the treatment of allergic inflammation diseases which has still no cure to this date. The inventors now surprisingly established that gliptins are inhibitors of intracellular MALT1 , opening up a whole new avenue for developing new treatments for allergic inflammatory disorders. Interfering in the NF-KB pathway by inhibiting MALT1 has been established by the inventors, resulting in silencing or killing of the immune cells (Th2) involved in the e.g. chronic allergic condition and / or a late-phase allergic condition of allergic inflammatory disorders.

[0227] A first aspect if the invention relates to a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating allergic inflammatory disease in a subject, preferably a human subject in need thereof.

[0228] An aspect of the invention relates to a therapeutic or prophylactic method of treating allergic inflammatory disease in a subject, preferably a human subject in need thereof, said method comprising the administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0229] An aspect of the invention relates to the use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of allergic inflammatory disease in a subject, preferably a human subject in need thereof.

[0230] Allergic inflammation

[0231] A type 2 immune response typically happens when your body senses certain kinds of infections or allergens and sends out specific type 2 immune cells to fight a battle against the intruders by creating inflammation. Type 2 immune diseases include among others Asthma, Atopic Dermatitis, Nasal polyps, eosinophilic esophagitis, prurigo nodularis and certain food allergies (Wang 2023).In certain embodiments, the allergic inflammatory disease is any one of chronic allergic condition and / or late-phase allergic condition. An embodiment is the pharmaceutical composition for use according to the invention, wherein the allergic inflammatory disease is a chronic allergic condition and / or a late-phase allergic condition.

[0232] Given its mechanism of action the gliptins, such as sitagliptin, is likely to be effective in chronic or late-phase allergic conditions, particularly those involving persistent Th2-driven inflammation. An embodiment is the pharmaceutical composition for use according to the invention, wherein the method results in inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside aberrant cells, preferably aberrant Th2 cells.

[0233] An embodiment is the pharmaceutical composition for use according to the invention, wherein the method results in inhibition of aberrant Th2 cells.

[0234] An embodiment is the pharmaceutical composition for use according to the invention, wherein the allergic inflammatory disease is mediated by or dependent on MALT1 activity.

[0235] An embodiment is the pharmaceutical composition for use according to the invention, wherein the method comprises the step of diagnosing a subject as suffering from or being at risk of suffering from allergic inflammatory disease mediated by or dependent on MALT1 activity.

[0236] In certain embodiments, the allergic inflammatory disease is allergic asthma, anaphylaxis, sinusitis, allergic rhinitis, eczema, hives, food allergy, chronic urticaria, atopic dermatitis and ocular allergic diseases. An embodiment is the pharmaceutical composition for use according to the invention, wherein the allergic inflammatory disease is selected from the group consisting of allergic asthma, anaphylaxis, sinusitis, allergic rhinitis, eczema, hives, food allergy, chronic urticaria, atopic dermatitis and ocular allergic diseases.

[0237] Conditions such as allergic asthma and perennial allergic rhinitis (e.g., due to dust mites or animal allergens) would be key areas to emphasize.

[0238] 1. Allergic Asthma

[0239] Allergic asthma involves Th2 cytokines (IL-4, IL-5, IL-13) and immune cells (eosinophils, T cells), which gliptins now can modulate via MALT1 inhibition. Chronic airway inflammation and remodelling in asthma will benefit from the potential tissue-modulating effects of the gliptins.

[0240] 2. Hay Fever (Allergic Rhinitis)

[0241] Hay fever is characterized by allergen-specific Th2-mediated responses (e.g., to specific pollens such as ragweed, birch, or grass). The ability of gliptins to reduce IL-4 and IL-13 could attenuate nasal inflammation and eosinophil recruitment. For pollen-allergy related allergic inflammation, target populations for gliptin treatment could include individuals allergic to common aeroallergens (e.g., grass pollen in temperate climates or birch pollen in Northern Europe). Based on the currently disclosed exemplifying data, the current invention now show that gliptin may be considered as part of a broader approach to managing seasonal allergic rhinitis.

[0242] 3. Food AllergyFood allergy involves mast cell activation, Th2 cytokines, and eosinophil recruitment in the gastrointestinal tract. Specific Foods: Based on the current invention, gliptins can be used for the treatment or prevention of allergic inflammation and related symptoms when food allergies such as allergies relating to common triggers like peanuts, tree nuts, shellfish, or milk are considered.

[0243] 4. Dust Mite Allergy

[0244] House dust mite allergens are a leading cause of perennial allergic rhinitis and asthma. Chronic exposure leads to airway inflammation and remodelling, which gliptin treatment can mitigate due to silencing of the MALT1 enzyme and therewith the NFKB pathway. For example, gliptins such as sitagliptin can be used for the treatment and for the managing of chronic allergic rhinitis or asthma in dust mite-sensitive individuals.

[0245] 5. Exposure to Animal Allergens

[0246] Allergens from cats, dogs, or rodents can trigger allergic rhinitis or asthma. The anti-inflammatory effects of gliptins, as now established by the inventors, may be used in treatments for allergic patients suffering from allergic inflammation (late-stage or chronic allergic inflammation) due to chronic exposure to animal allergens. For example, patients with persistent allergic inflammation due to household or occupational exposure to animal allergens can be selected for treatment with a gliptin.

[0247] MALT1

[0248] The Mucosa-associated lymphoid tissue lymphoma translocation 1 (MALT1) is a paracaspase and a central signalling protein which forms part of the intracellular CBM (CARMA / CARD-BCL10-MALT1) signalling complex downstream of receptors containing immunoreceptor tyrosine-based activation motif (ITAMs) as well as a subset of G-protein-coupled receptors (GPCRs) and receptor tyrosine kinases found in both hematopoietic and non-hematopoietic cell types. Following receptor activation, the CBM complex activates downstream signalling events via the scaffolding and protease functions of MALT1 that regulates key cellular processes, including lymphocyte development and activation of cells of the innate and adaptive immune system. Following TCR or BCR triggering, MALT 1 is required for activation of the canonical NF-kB pathway MALT1 partners with BCL10 and CARMA1 to form the CARMA1-BCL10-MALT1 signalosome complex (CBM), where MALT1 has been thought to act as a major protein scaffold that recruits downstream effector proteins to activate NF-kB signaling. MALT1 also plays a role in the activation of the non-canonical NF-kB pathway upon B cell activating factor receptor (BAFF-R) stimulation, a pathway that is needed for survival of marginal zone (MZ) B cells.

[0249] Thus, MALT1 has dual role: i) scaffold activity for the assembly of CBM complex and ii) proteolytic activity which is thought to promote NF-KB activation through the cleavage of the deubiquitinating enzyme A20 and of the NF-KB subunit RelB, CYLD and HOIL-1. Moreover, MALT1 possesses the capacity of autocleavage, which is an additional necessary step in NF-KB activation. MALT1 -dependent RelB cleavage results in its proteasomal degradation, thereby lowering total levels of RelB in activated B- and T cells. Through cleavage of RelB and A20, MALT1 thus expands the amplitude and duration of the NF-KB response in an IKK-independent manner. Interestingly, MALT1also promotes NF-KB activation by its autoprocessing after Arg 149. The exact consequences of MALT1 autoprocessing remain unclear, but include an effect on NF-KB target gene expression downstream of nuclear NF-KB accumulation.

[0250] The role of the paracaspase activity of MALT1 has been studied in vivo using genetically modified knock-in (MALT1-PD) mice expressing an inactive form of the MALT1 protease.

[0251] MALT1 inhibitors

[0252] MALT 1 ’s enzymatic activity has stimulated efforts to develop specific MALT 1 inhibitors for research and therapeutic purposes. The first MALT1 inhibitor identified was z-VRPR-fmk, a modified tetrapeptide based on the optimal substrate Val-Arg-Pro-Arg of the Arabidopsis thaliana metacaspase AtmC9, conjugated to fluoromethyl ketone (fmk). This modified peptide irreversibly blocked MALT1 protease activity in an in vitro cleavage assay using recombinant MALT1 in a dose-dependent manner. It furthermore efficiently inhibited T cell activation and IL-2 secretion in JurkatT cells and in human antigen specific CTLs. Nagel and colleagues have identified three phenothiazine derivatives (mepazine, thioridazine, and promazine) as highly specific, noncompetitive and reversible MALT1 inhibitors. A concurrent study by Fontan and colleagues has identified the compound MI-2 as a selective MALT1 inhibitor. In contrast to phenothiazine derivatives, MI-2 is a covalent and irreversible inhibitor that acts on the catalytic centre of MALT1.

[0253] It has been shown that reversible and irreversible inhibitors of MALT1 can act differently on autoimmune cells. The therapeutic potential of targeting MALT1 , although promising, has been hindered by concerns regarding the impact of MALT inhibition on Treg homeostasis.

[0254] MALT1 as target for allergic inflammation

[0255] MALT1 is a signalling protein that plays a key role in immunity, inflammation, and lymphoid malignancies. For a long time MALT1 was believed to function as a scaffold protein, providing an assembly platform for other signalling proteins. This view changed dramatically when MALT1 was also found to have proteolytic activity and a capacity to fine-tune immune responses. Preclinical studies have supported the belief that MALT1 is a suitable therapeutic target in allergic inflammation diseases.

[0256] Phenothiazines inhibit MALT1 Activity and IL-2 Induction in T Cells

[0257] It is known that phenothiazines inhibit MALT1 Activity and IL-2 Induction in T Cells. Derivatives of medicinal active phenothiazines, including mepazine, have been identified as small molecule inhibitors of the MALT1 protease (Nagel 2012). These phenothiazines selectively inhibit cleavage activity of recombinant and cellular MALT1 by a noncompetitive mechanism. As MALT1 protease activity is required for T cell responses, the effects on MALT1 activity and IL-2 production were tested in T cells. A MALT1 cleavage assay was used after immunoprecipitation (IP) of the protein from Jurkat T cells. Cells were left untreated or incubated for 3 hr with 10 mM of mepazine or thioridazine and subsequently left unstimulated or stimulated with anti-CD3 / CD28. MALT1 protease activity was almost undetectable in the absence of stimulation and peaked at 30-60 min after CD3 / CD28 treatment. Addition of either mepazine or thioridazine resulted in a strong reduction of MALT1 protease activityin stimulated Jurkat T cells at all time points.

[0258] Mepazine has been evaluated as an antipsychotic and tranquilizing drug under the brand name Pacatai in the late 1950s and early 1960s. Whereas some clinical investigations provided evidence for antipsychotic effects, other studies failed to do so (Sarwer-Foner and Koranyi, 1957; Whittier et al., 1960). Some side effects were reported after mepazine treatment, including a reduction of asthma attacks, indicating potential immunosuppressive activity (Sarwer-Foner and Koranyi, 1957).

[0259] While the phenothiazine-derivatives, mepazine and (S)-mepazine, are the first known allosteric MALT1 inhibitors with medium potency, the compounds are not highly selective for MALT1 and antagonize other proteins such as G protein coupled receptors (GPCR). Thus, mepazine may affect cellular functions independent of MALT1 and, for instance, mepazine impairs CYP450 isoenzymes. Mepazine is doing bad on CYP450 activity (almost full inhibition) by interfering due to CYP2D6 inhibition and mepazine is almost completely silencing CYP450 metabolic activity due to interfering with CYP2D6. Moreover, despite the MALT1 activity, this compound shows some side effects like tardive dyskinesia which limits its clinical study (Liang et al. 2021a).

[0260] Cytochrome (CYP) 450 isoenzymes control

[0261] Cytochrome (CYP) 450 isoenzymes are the basic enzymes involved in Phase I biotransformation. The most important role in biotransformation belongs to CYP3A4, CYP2D6, CYP2C9, CYP2C19 and CYP1A2. Inhibition and induction of CYP isoenzymes caused by drugs are important and clinically relevant pharmacokinetic mechanisms of drug interaction and for assessing risks of drugs.

[0262] Gli ptins

[0263] Surprisingly, the inventors have identified gliptins as MALT1 inhibitors. Gliptins have been found to bind to the allosteric site of MALT1 which is located intracellularly.

[0264] Gliptins are a class of pharmaceutical compounds which are the inhibitors of dipeptidyl peptidase 4, also known as DPP-4 inhibitors. Now, the inventors identify a new target for the gliptins which is MALT 1 , an intracellular signalling protein that is widely expressed and known to be implicated in innate [natural killer cells (NK), dendritic cells (DC), and mast cells] and adaptive immune cells (T cells and B cells), where it signals proinflammatory gene expression downstream of several cell-surface receptors.

[0265] Gliptins are known to bind the extracellular domain of DPP4, also known as CD26, which is a surface T cell activation antigen and has been shown to have DPP4 enzymatic activity, cleaving-off amino-terminal dipeptides with either L-proline or L-alanine at the penultimate position. Therefore, it plays a major role in glucose metabolism by N-terminal truncation and inactivation of the incretins glucagon-like peptide-1 (GLP) and gastric inhibitory protein (GIP). However, a large number of studies demonstrate clearly that CD26 / DPP4 also plays an integral role in the immune system, particularly in T cell activation.

[0266] The gliptins having DPP4 inhibition activity known include Sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin, Alogliptin Teneligliptin, Trelagliptin, Omarigliptin, Evogliptin,Gosogliptin, Retagliptin Dutogliptin, Neogliptin, Denagliptin, Cofrogliptin, Fotagliptin, Prusogliptin, Cetagliptin.

[0267] In certain embodiments of the invention, the gliptin is any one of Sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin, Alogliptin Teneligliptin, Trelagliptin, Omarigliptin, Evogliptin, Gosogliptin, Retagliptin Dutogliptin, Neogliptin, Denagliptin, Cofrogliptin, Fotagliptin, Prusogliptin, Cetagliptin.

[0268] In certain (preferred) embodiments of the invention, the gliptin is any one of Sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin, Alogliptin, Teneligliptin, Trelagliptin, Omarigliptin, Evogliptin, Gosogliptin, Retagliptin

[0269] In certain (preferred) embodiments of the invention, the gliptin is any one of Sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin, Alogliptin.

[0270] In certain (preferred) embodiments of the invention, the gliptin is any one of Sitagliptin, Linagliptin, Alogliptin.

[0271] An embodiment is the pharmaceutical composition for use according to the invention, wherein the gliptin is Sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin, Alogliptin, Teneligliptin, Trelagliptin, Omarigliptin, Evogliptin, Gosogliptin, Retagliptin, Denagliptin, Cofrogliptin, Fotagliptin, Prusogliptin, Dutogliptin, Neogliptin or Cetagliptin, preferably wherein the gliptin is Sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin, Alogliptin, Teneligliptin, Trelagliptin, Omarigliptin, Evogliptin, Gosogliptin, Retagliptin, Denagliptin, Cofrogliptin, Fotagliptin, Prusogliptin, more preferably wherein the gliptin is sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin or Alogliptin, more preferably wherein the gliptin is Sitagliptin, Linagliptin, Vildagliptin, Saxagliptin or Alogliptin, most preferably wherein the gliptin is sitagliptin.

[0272] An embodiment is the pharmaceutical composition for use according to the invention, wherein the gliptin is Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin, Alogliptin, Teneligliptin, Trelagliptin, Omarigliptin, Evogliptin, Gosogliptin, Retagliptin, Dutogliptin, Neogliptin, Denagliptin, Cofrogliptin, Fotagliptin, Prusogliptin, or Cetagliptin, preferably wherein the gliptin is Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin or Alogliptin, more preferably wherein the gliptin is Vildagliptin, Saxagliptin, Linagliptin or Alogliptin.

[0273] An embodiment is the pharmaceutical composition for use according to the invention, wherein the gliptin is not Sitagliptin.

[0274] In certain (preferred) embodiments of the invention, the gliptin is Sitagliptin.

[0275] An embodiment is the pharmaceutical composition for use according to the invention, wherein the gliptin is an inhibitor of MALT1 , preferably an allosteric inhibitor of MALT1.

[0276] Structures of gliptins are shown below:

[0277]

[0278]

[0279]

[0280]

[0281]

[0282] In one embodiment, the gliptin is sitagliptin.

[0283] In another embodiment, the gliptin is vildagliptin.

[0284] In yet another embodiment, the gliptin is saxagliptin. In a further embodiment, the gliptin is linagliptin.

[0285] In another embodiment, the gliptin is gemigliptin.

[0286] In yet another embodiment, the gliptin is anagliptin. In a further embodiment, the gliptin is alogliptin.

[0287] In one embodiment, the gliptin is retagliptin.

[0288] In another embodiment, the gliptin is evogliptin.

[0289] In yet another embodiment, the gliptin is trelagliptin. In a further embodiment, the gliptin is teneligliptin. In another embodiment, the gliptin is gosogliptin.

[0290] In yet another embodiment, the gliptin is omarigliptin.In silico modelling MALT1 -docking

[0291] Furthermore, in silico modelling indicate that these gliptins (all) show an interaction with allosteric MALT1 -target site, see Example 3.

[0292] Sitagliptin

[0293] Sitagliptin (C16H15F6N5O; (3R)-3-amino-1-[3-(trifluoromethyl)-6,8-dihydro-5H-[1 ,2,4]triazolo[4,3-a]pyrazin-7-yl]-4-(2,4,5-trifluorophenyl)butan-1-one) is the first antidiabetic agent from the class of dipeptidyl peptidase-4 (DPP-4) enzyme inhibitors. It increases the amount of circulating incretins, which stimulate insulin secretion and inhibit glucose production (Choy et al, 2007).

[0294] DPP-4 is an enzyme that breaks down incretin hormones such as glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide-1 (GIP) (Plosker, 2014). By inhibiting DPP-4, sitagliptin results in improved glucose control. Additionally, sitagliptin decreases the rate at which glucose is produced by the liver. This decreased hepatic glucose production contributes further to improved glycaemic control in patients with type- 2 diabetes (Katsuno et al, 2013; Sakura et al, 2016).

[0295] Several preclinical studies have also shown that DPP-4 inhibitors can improve endothelial function, reduce oxidative stress, and attenuate inflammation, all of which are key factors in the pathogenesis of cardiovascular diseases (Hanssen and Jandeleit-Dahm, 2019; Liu et al, 2019; Chikata et al, 2022). Clinical trials have also demonstrated that sitagliptin can improve left ventricular function and reduce circulating biomarkers of inflammation and oxidative stress in patients with heart failure and preserved ejection fraction (Zannad et al, 2015). The underlying mechanism for these effects is related to the inhibition of DPP-4, which leads to increased levels of GLP-1 and other vasoactive peptides that enhance endothelial function and reduce inflammation (Kim et al, 2014).

[0296] Administration

[0297] An embodiment is the pharmaceutical composition for use according to the invention, wherein the method comprises oral administration of the pharmaceutical composition.

[0298] Dose regimen

[0299] In embodiments, is the dose of gliptin, preferably the once daily dose of the gliptin is preferably less than 1000 mg, preferable less than 500 mg, less than 250 mg, less than 200 mg, less than 150 mg, or egual to or less than 100 mg, even more preferably, egual to or less than 50 mg, 25 mg, 20 mg, 12,5 mg, 12 mg, 10 mg, 6,25 mg, 5 mg or 2,5 mg. For some gliptins, preferred (once daily) doses are listed in Table B.

[0300] TABLE B. Examples of doses for a series of gliptins for use according to the invention

[0301]

[0302]

[0303] In Summary:

[0304] An aspect of the disclosure relates to a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating a MALT-1 mediated disease selected from: allergic inflammatory disease.

[0305] An embodiment is the pharmaceutical composition for use, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin or cetagliptin, preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, more preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin or alogliptin, most preferably wherein the gliptin is sitagliptin.

[0306] An embodiment is the pharmaceutical composition for use, wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, or cetagliptin, preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin or alogliptin.

[0307] An embodiment is the pharmaceutical composition for use, wherein the gliptin is not sitagliptin. An embodiment is the pharmaceutical composition for use, wherein the gliptin is an inhibitor of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1), preferably an allosteric inhibitor of MALT1.

[0308] An embodiment is the pharmaceutical composition for use, wherein the therapeutic effect of the gliptin comprises and / or results from inhibition of MALT1 , preferably wherein the inhibition of MALT1 is an allosteric inhibition resulting from binding of the gliptin to an allosteric site within MALT1.An embodiment is the pharmaceutical composition for use, wherein the method comprises oral administration of the pharmaceutical composition.

[0309] An embodiment is the pharmaceutical composition for use, wherein the method results in inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside aberrant cells, preferably aberrant Th2 cells.

[0310] An embodiment is the pharmaceutical composition for use, wherein the allergic inflammatory disease is a chronic allergic condition and / or a late-phase allergic condition.

[0311] An embodiment is the pharmaceutical composition for use, wherein the allergic inflammatory disease is selected from the group consisting of allergic asthma, anaphylaxis, sinusitis, allergic rhinitis, eczema, hives, food allergy, chronic urticaria, atopic dermatitis and ocular allergic diseases.

[0312] An embodiment is the pharmaceutical composition for use, wherein the allergic inflammatory disease is mediated by or dependent on MALT1 activity.

[0313] An embodiment is the pharmaceutical composition for use, wherein the method comprises the step of diagnosing a subject as suffering from the allergic inflammatory disease, or being at risk of suffering from the allergic inflammatory disease mediated by or dependent on MALT1 activity.

[0314] An embodiment is the pharmaceutical composition for use, wherein the treatment is followed by radiotherapy and / or chemotherapy.

[0315] An embodiment is the pharmaceutical composition for use, wherein MALT1 expression status, and optionally DPP-4 expression status, comprises RNA expression analysis and / or protein expression analysis, preferably by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and / or immunoassay.

[0316] An embodiment is the pharmaceutical composition for use, wherein the method comprises the oral administration of the gliptin, pharmaceutically acceptable salt thereof or hydrate thereof, at a dose of 0.01 mg to 1000 mg per day.

[0317] An embodiment is the pharmaceutical composition for use, wherein the pharmaceutical composition is provided in an oral unit dosage form, preferably in the form of a tablet, comprising the gliptin, the pharmaceutically acceptable salt thereof or hydrate thereof, in an amount of 0.01 mg to 1000 mg.

[0318] An aspect of the disclosure relates to a therapeutic or prophylactic method of treating a MALT-1 mediated disease selected from allergic inflammatory disease, in a subject, preferably human subject, in need thereof, said method comprising the step of administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0319] An aspect of the disclosure relates to a use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of a MALT-1 mediated disease selected from allergic inflammatory disease, in a subject, preferably human subject, in need thereof.AUTOIMMUNE DISEASES, PREFERABLY MS

[0320] As said, the present invention relates to the unexpected finding of the inventors that gliptins, which are known to be DPP-4 inhibitors, can bind to an allosteric site on MALT1 and inhibit the same reversibly. This surprising and unexpected finding leads to a further novel use of the gliptins for the treatment of autoimmune diseases, particularly for the treatment and / or prevention of multiple sclerosis which has still no cure to this date.

[0321] As indicated herein before, the following aspects of the invention are provided:

[0322] A therapeutic or prophylactic method of treating MS in a human subject in need thereof, said method comprising the administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0323] A pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating multiple sclerosis (MS) in a human subject in need thereof.

[0324] Use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of MS in a human subject in need thereof.

[0325] In embodiments of the invention, the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin or cetagliptin, preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, more preferably the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin or alogliptin, most preferably the gliptin is sitagliptin.

[0326] In a further embodiment of the invention the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, or cetagliptin, preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin or alogliptin.

[0327] In certain preferred embodiment of the invention the gliptin is not sitagliptin.

[0328] An overview for different types of gliptins, their formulas and synthesis pathways can be found in Kushwaha (2014), which is herewith incorporated by reference.

[0329] Structures of gliptins are shown below:

[0330]

[0331]

[0332]

[0333]

[0334]

[0335] In a specific embodiment of the invention, the gliptin is vildagliptin. In a specific embodiment of the invention the gliptin is saxagliptin. In a specific embodiment of the invention the gliptin is linagliptin. In a specific embodiment of the invention the gliptin is gemigliptin. In a specific embodiment of the invention the gliptin is anagliptin. In a specific embodiment of the invention the gliptin is alogliptin. In a specific embodiment of the invention the gliptin is teneligliptin. In a specific embodiment of the invention the gliptin is trelagliptin. In a specific embodiment of the invention the gliptin is omarigliptin. In a specific embodiment of the invention the gliptin is evogliptin. In a specific embodiment of the invention the gliptin is gosogliptin. In a specific embodiment of the invention the gliptin is retagliptin. In a specific embodiment of the invention the gliptin is denagliptin.In a specific embodiment of the invention the gliptin is cofrogliptin.

[0336] In a specific embodiment of the invention the gliptin is fotagliptin.

[0337] In a specific embodiment of the invention the gliptin is prusogliptin.

[0338] In a specific embodiment of the invention the gliptin is dutogliptin.

[0339] In a specific embodiment of the invention the gliptin is neogliptin.

[0340] In a specific embodiment of the invention the gliptin is cetagliptin.

[0341] In a further embodiment of the invention, the gliptin is an inhibitor of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1), preferably an allosteric inhibitor of MALT1.

[0342] In particularly preferred embodiments ofthe invention, the method of treating MS comprises oral administration ofthe pharmaceutical composition.

[0343] In an embodiment ofthe invention, the method results in inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside aberrant cells, preferably aberrant T helper 1 (Th1), T helper 2 (Th2), T helper 9 (Th9), T helper 17 (Th17), T helper 22 (Th22), and follicular helper T cells (Tfh), more preferably Th1 cells, Th17 cells.

[0344] In an embodiment of the invention, the MS is clinically isolated syndrome (CIS), relapsingremitting MS (RRMS), secondary progressive MS (SPMS), primary progressive MS (PPMS) or progressive relapsing MS (PRMS).

[0345] In an embodiment of the invention, the MS is at pre-symptomatic stage, clinically isolated syndrome stage, relapsing-remitting stage, or at secondary progressive stage.

[0346] In an embodiment ofthe invention, the MS is mediated by or dependent on MALT1 activity (i.e ., the MS is a MALT-1 mediated disease or a MALT-1 involving disease).

[0347] In an embodiment ofthe invention, the MS is mediated by or dependent on MALT1 activity which induces a (TCR)-mediated NF-KB activation.

[0348] In an embodiment of the invention, the inhibition of MALT1 comprises or results in an inhibition of cell survival in a (TCR)-mediated NF-KB activated cell.

[0349] In an embodiment of the invention, the inhibition of MALT1 comprises or results in an inhibition of cell metabolism in a (TCR)-mediated NF-KB activated cell.

[0350] In an embodiment of the invention, the method comprises the step of diagnosing a subject as suffering from or being at risk of suffering from MS mediated by or dependent on MALT1 activity.

[0351] In an embodiment of the invention, the method comprises the oral administration of the gliptin, pharmaceutically acceptable salt thereof or hydrate thereof, at a dose of 0.01 mg to 1000 mg per day.

[0352] In an embodiment of the invention, the method comprises the oral administration of the gliptin, pharmaceutically acceptable salt thereof or hydrate thereof, at a dose less than 1000 mg, preferably less than 500 mg, less than 250 mg, less than 200 mg, less than 150 mg, or less than 100 mg per day.

[0353] In an embodiment of the invention, the pharmaceutical composition is provided in an oral unit dosage form, preferably in the form of a tablet, comprising the gliptin, the pharmaceutically acceptable salt thereof or hydrate thereof, in an amount of 0.01 mg to 1000 mg.

[0354] In further embodiments, the pharmaceutical composition is provided in an oral unit dosage form, preferably in the form of a tablet, comprising the gliptin, the pharmaceutically acceptable salt thereof or hydrate thereof, less than 1000 mg, preferably less than 500 mg, less than 250 mg, less than 200 mg,less than 150 mg, or equal to or less than 100 mg, even more preferably, equal to or less than 50 mg, 25 mg, 20 mg, 12,5 mg, 12 mg, 10 mg, 6,25 mg, 5 mg or 2,5 mg. For some gliptins, preferred (once daily) doses are listed in Table A.

[0355] TABLE A. Examples of doses for a series of gliptins for use according to the invention

[0356]

[0357] The present invention further relates to a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof.

[0358] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is selected from sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, cetagliptin.

[0359] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is selected from sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin.

[0360] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is selected from sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin.

[0361] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is selected from sitagliptin, linagliptin, alogliptin.

[0362] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is sitagliptin.An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is selected from vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, cetagliptin.

[0363] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is selected from vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin.

[0364] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is selected from vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin.

[0365] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is selected from linagliptin, alogliptin.

[0366] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is not sitagliptin.

[0367] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is vildagliptin.

[0368] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is saxagliptin.

[0369] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is linagliptin.

[0370] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is gemigliptin.

[0371] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is anagliptin.

[0372] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is alogliptin.

[0373] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is teneligliptin.

[0374] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is trelagliptin.

[0375] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is omarigliptin.An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is evogliptin.

[0376] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is gosogliptin.

[0377] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is retagliptin.

[0378] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is dutogliptin.

[0379] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is neogliptin.

[0380] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is denagliptin.

[0381] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is cofrog li ptin.

[0382] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is fotagliptin.

[0383] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is prusogliptin.

[0384] An embodiment is a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the gliptin is cetagliptin.

[0385] The present invention further relates to a gliptin or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of multiple sclerosis (MS) in a subject in need thereof, wherein the method comprises administering to a subject, an effective amount of the gliptin or the pharmaceutically acceptable salt thereof.

[0386] As will be explained in detail below, several inflammatory cytokines are involved in the initiation, progress, treatment and / or prevention of MS. The inventors have established that gliptins allosterically inhibit MALT1 and, via that mechanism will have utility in a therapeutic or prophylactic method of treating MS in a human subject in need thereof.

[0387] In line with these findings, the present methods comprise or result in inhibition of mucosa-associated lymphoid tissue lymphoma translocation 1 (MALT1).

[0388] Multiple Sclerosis (MS)

[0389] Four clinical courses of MS have been identified: relapsing-remitting MS (RRMS), secondary progressive MS (SPMS), primary progressive MS (PPMS) and progressive relapsing MS (PRMS). Approximately 85% of the patients display RRMS which is characterized by the occurrence of relapses at irregular intervals with complete or incomplete neurological recovery. Almost 10-15% of patientspresent with PPMS is another course of MS which is characterized by disease progression from the onset, resulting in gradual, progressive and permanent neurological deficits for>1 year without relapses. A rare clinical course is PRMS characterized by progressive disease from the onset, with acute relapses (with or without full clinical recovery) and periods of continuing progression between relapses. In addition to these phenotypes a further stage and / or episode is included. Clinically isolated syndrome (CIS) denotes those patients whose first clinical presentation has characteristics of inflammatory demyelination that could be MS but who do not fulfil its diagnostic criteria. Within each subtype, it is possible to diagnose and classify the disease whether it is active or not active which are defined by the occurrence of relapses or lesions detected using MRL Another important modifier of the progressive stages is the inclusion of whether disability has progressed over a given time period.

[0390] In line with the above, in an embodiment, the present methods result in inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside aberrant cells, preferably aberrant T helper 1 (Th1) cells, Th17 cells.

[0391] In an embodiment, the MS treated in accordance with the invention is clinically isolated syndrome (CIS), relapsing-remitting MS (RRMS), secondary progressive MS (SPMS), primary progressive MS (PPMS) or progressive relapsing MS (PRMS).

[0392] In an embodiment, the MS is at pre-symptomatic, clinically isolated syndrome, relapsingremitting, or secondary progressive stage.

[0393] In an embodiment, the therapeutic or prophylactic method is a method of treating Th17 cell mediated disease and / or MALT1 mediated disease.

[0394] A further embodiment of the invention, the therapeutic or prophylactic method, is a method of treating Th17 cell mediated disease and / or MALT1 mediated disease, wherein the disease is rheumatoid arthritis, systemic lupus erythematosus, systemic sclerosis, autoimmune arthritis, inflammatory colitis.

[0395] Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system (CNS) where immunopathology is thought to be mediated by myelin-reactive CD4+T helper (Th) cells. The Th cells most commonly implicated in the pathogenesis of the disease are of Th1 and Th17 lineage, which are defined by the production of interferon-y and interleukin-17, respectively. Importantly, all Th17 cells carry the C-C chemokine receptor 6 (CCR6, CD196), express the transcription factor retinoic acid-related orphan receptor C (RORC) as well as different amounts of IL-17. 1 ntriguingly , while regulatory T cells (Tregs, CD4+CD25hi9hFoxP3+) protect from and Th17 cells cause autoimmune disorders, Tregs can acquire Th17 characteristics.

[0396] The pathogenicity of Th17 cells can be distinguished by the expression of different cell surface markers including CXCR3 and CCR4. For example, the classical Th17 cells (CD4+CCR6+CXCR3_CCR4+) produce high levels of IL-17, an interleukin that has the ability to impair the blood-brain-barrier (BBB) and thus makes them highly encephalitogenic. The BBB breakdown is partly induced by the formation of reactive oxygen induced by IL-17. Moreover, human endothelial cells of pwMS express high levels of IL-17 receptors, possibly facilitating CNS transmigration independent of the BBB disruption. The transcription factor RORC induces IL-17 (as well as CCR6) production and high expression of RORC therefore defines this cell type. After dysregulation of the cerebral tight junctions, CNS infiltration is facilitated by the dense expression of the CCR6 on the cell surface. CCR6 interactswith CCL20, which is highly expressed on sites of inflammation, e.g. on the epithelium of the choroid plexus. IL-17 further amplifies this process by inducing CCL20 expression. Neuronal damage is promoted as a result of apoptotic granzyme B released by myelin specific Thi? cells and indirectly, as IL-17 stimulates CNS-resident cells to produce inflammatory mediators. Some of these cells, over their lifespan, eventually down-regulate IL-17 and produce high amounts of IFN-y, the signature cytokine of Th1 cells, and GM-CSF instead. This phenotype conversion gives rise to a new subset of Th cells called Th17.1 (CD4+CCR6+CXCR3+CCR4) promoted by IL-1 and IL-23. Interferon-y (IFN-y) and granulocyte macrophage colony-stimulating factor (GM-CSF) stimulate macrophages, neutrophils and dendritic cells augmenting inflammation through components of the innate immune system. IFN-y activates also brain endothelial cells, which hence upregulate the expression of adhesion molecules such as ICAM-1. Th17.1 subgroup share common inflammatory and pathogenic features of both Th1 and Th17 cells and further disintegrates the BBB and eases lymphocyte transmigration. CNS infiltration is additionally promoted by the high expression of very late antigen 4 (VLA-4) on the surface of TH17.1 cells. In fact, they more efficiently transmigrate the BBB than single (IL-17 or IFN-y) producers. Moreover, TH17.1 cells show higher proliferation rates upon activation as compared to the classical Th17 cells and induce the expression of multidrug-resistance 1 (MDR-1) protein, potentially protecting them from therapeutic agents. In addition, they appear to be resistant to suppression by Tregs. Taken together, Th17.1 cells possess a series of features, which make them particularly proinflammatory. They have been linked to several autoimmune disorders including diabetes mellitus type I, autoimmune arthritis, inflammatory colitis and importantly multiple sclerosis. A further subgroup of T cells are defined by the presence of surface marker CXCR3 and CCR4, hence called double positive in addition to CD4+CCR6+. They produce low amounts of INF-y and GM-CSF but intermediate IL-17 quantities. And last, CD20+CD4+T cells also produce IL-17 and express CCR6. They represent particularly proinflammatory cell clones with high transmigrational potential and have been linked to MS severity. Notably, abundant CD20+ T cells were found in active MS lesions, which expressed IL-17 and INF-y.

[0397] Other cell types that have been involved in the pathology of MS include further subset of T cells comprising Th2, Th17, Th22, and follicular helper T cells (Tfh).

[0398] In view of the above, it is clear that the severity and progress of the disease are dependent on the differentiated T cells and the excretion of cytokines including IL-17 and INF-y.

[0399] NF-KB plays a significant role not only as a mediator of the inflammatory process in peripheral immune cells but also in microglia and astrocytes, making it important in the progression of MS. The differentiation of T cells into regulatory T cells (Tregs) requires the NF-KB subunit c-Rel, indicating that NF-KB could be a potential therapeutic target for MS. Furthermore, antigen-presenting cells (APCs) depend on c-Rel to produce IL-12 and IL-23, two cytokines that promote the differentiation of ThO into Th1 or Th17 cells, respectively. Th17 cells secrete granulocyte-macrophage colony-stimulating factor (GM-CSF), which plays a crucial role in promoting the progression of MS. The activation of the NF-KB pathway is required for astrocytes to create a local inflammatory environment by inducing the secretion of inflammatory cytokines and chemokines. This inflammatory response can result in local tissue damage and attract additional inflammatory cells. Conversely, inhibiting the NF-KB pathway in astrocytes leads to the recruitment of CD8 + CD122+ regulatory T cells, thereby alleviatingMS. Similarly, the activation of NF-KB in microglia or infiltrating macrophages may also influence the production of inflammatory cytokines and chemokine. Persistent activation of the NF-KB pathway is frequently implicated besides MS in other inflammatory conditions like rheumatoid arthritis, inflammatory bowel disease, and asthma.

[0400] To conclude, the balance of the cytokines excreted in the microenvironment, the modulation and differentiation of naive T cells into T regulatory cells (Tregs) and Th1 / Th17 cells are crucial parameters implicated in the progress and treatment of MS.

[0401] To study the pathology of T cells in MS, several animal and / or cell models can be used. One of the animal models used in MS is the cuprizone mouse models, as defined in Praet et al., which are characterized by the intact blood-brain barrier. However, this model is not suitable to observe the T cell regulation because one of the hallmark of MS is the broken down or leaky blood-brain-barrier. The cuprizone mouse model lacks obviously this hallmark of MS, therefore does not allow to identify and / or determine the pathology of T cells.

[0402] Regarding cell models, Jurkat cells are an appropriate model for studying aspects of multiple sclerosis (MS), especially the immune-mediated inflammatory mechanisms. Jurkat cells are immortalized line of human T lymphocyte cells that are used to study acute T cell leukemia, T cell signaling, and the expression of various chemokine receptors susceptible to viral entry, particularly HIV. They are widely used in immunology research to study T-cell receptor (TCR) signalling, cytokine production, and other pathways responsible for initiating MS. Inducing Jurkat cells with the relevant cytokines and antibodies which mimics the activation of autoreactive CD4+T-helper cells including Th1 and Th 17 provides a suitable model for T cell pathology in MS. Therefore, Jurkat cells are ideal for studying TCR-mediated pathways, including activation of transcription factors like NF-KB, NFAT, and AP-1 , which drive the production of pro-inflammatory cytokines such as IL-2, IFN-y, and TNF-a. These cells are a robust system for analysing cytokine production in response to stimulation e.g., with anti-CD3 / CD28 antibodies which is very relevant to analyze dysregulated cytokine profiles (e.g., elevated IL-17, IFN-y, and IL-6) playing a central role in MS. Therefore, Jurkat cells, being easy to culture and highly responsive to TCR stimulation, provide an accessible, reproducible, and cost-effective platform for early-stage mechanistic studies.

[0403] MALT1

[0404] The Mucosa-associated lymphoid tissue lymphoma translocation 1 (MALT1) is a paracaspase and a central signalling protein which forms part of the intracellular CBM (CARMA / CARD-BCL10-MALT1) signalling complex downstream of receptors containing immunoreceptor tyrosine-based activation motif (ITAMs) as well as a subset of G-protein-coupled receptors (GPCRs) and receptor tyrosine kinases found in both hematopoietic and non-hematopoietic cell types. Following receptor activation, the CBM complex activates downstream signalling events via the scaffolding and protease functions of MALT1 that regulates key cellular processes, including lymphocyte development and activation of cells of the innate and adaptive immune system. Following TCR or BCR triggering, MALT1 is required for activation of the canonical NF-kB pathway MALT1 partners with BCL10 and CARMA1 to form the CARMA1-BCL10-MALT1 signalosome complex (CBM), where MALT1 has been thought to act as a major protein scaffold that recruits downstream effector proteins to activate NF-kB signalling.MALT1 also plays a role in the activation of the non-canonical NF-kB pathway upon B cell activating factor receptor (BAFF-R) stimulation, a pathway that is needed for survival of marginal zone (MZ) B cells.

[0405] Thus, MALT1 has dual role: i) scaffold activity for the assembly of CBM complex and ii) proteolytic activity which is thought to promote NF-KB activation through the cleavage of the deubiquitinating enzyme A20 and of the NF-KB subunit RelB, CYLD and HOIL-1. Moreover, MALT1 possesses the capacity of autocleavage, which is an additional necessary step in NF-KB activation. MALT1 -dependent RelB cleavage results in its proteasomal degradation, thereby lowering total levels of RelB in activated B- and T cells. Through cleavage of RelB and A20, MALT1 thus expands the amplitude and duration of the NF-KB response in an IKK-independent manner. Interestingly, MALT1 also promotes NF-KB activation by its autoprocessing after Arg 149. The exact consequences of MALT1 autoprocessing remain unclear, but include an effect on NF-KB target gene expression downstream of nuclear NF-KB accumulation.

[0406] The role of the paracaspase activity of MALT1 has been studied in vivo using genetically modified knock-in (MALT1-PD) mice expressing an inactive form of the MALT1 protease. Somewhat surprisingly, several independent studies have reported development of spontaneous lethal autoimmune inflammation, such as multi-organ inflammation, gastritis and ataxia, in MALT1-PDmice.

[0407] MALT1 inhibitors

[0408] MALT 1 ’s enzymatic activity has stimulated efforts to develop specific MALT 1 inhibitors for research and therapeutic purposes. The first MALT1 inhibitor identified was z-VRPR-fmk, a modified tetrapeptide based on the optimal substrate Val-Arg-Pro-Arg of the Arabidopsis thaliana metacaspase AtmC9, conjugated to fluoromethyl ketone (fmk). This modified peptide irreversibly blocked MALT1 protease activity in an in vitro cleavage assay using recombinant MALT1 in a dose-dependent manner. It furthermore efficiently inhibited T cell activation and IL-2 secretion in JurkatT cells and in human antigen specific CTLs.

[0409] Nagel and colleagues have identified three phenothiazine derivatives (mepazine, thioridazine, and promazine) as highly specific, noncompetitive and reversible MALT1 inhibitors. A concurrent study by Fontan and colleagues has identified the compound MI-2 as a selective MALT1 inhibitor. In contrast to phenothiazine derivatives, MI-2 is a covalent and irreversible inhibitor that acts on the catalytic centre of MALT1 .

[0410] It has been shown that reversible and irreversible inhibitors of MALT1 can act differently on autoimmune cells. The therapeutic potential of targeting MALT1 , although promising, has been hindered by concerns regarding the impact of MALT inhibition on Treg homeostasis.

[0411] For example, while the phenothiazine-derivatives, mepazine and (S)-mepazine, are the first known allosteric MALT1 inhibitors with medium potency, the compounds are not highly selective for MALT1 and antagonize other proteins such as G protein coupled receptors (GPCR). Thus, mepazine may affect cellular functions independent of MALT1 and, for instance, mepazine impairs RANK-induced osteoclastogenesis independent of its MALT1 inhibitory function. Furthermore, despite this good inhibitory activity, this compound shows some side effects like tardive dyskinesia which limits its clinical study. Additionally, mepazine also interferes in the CYP450 and CYP2D4 pathway.On the other hand, in MS and its main animal model, experimental autoimmune encephalomyelitis (EAE), which are characterized by myelin-specific autoreactive T cells, a MALT1 inhibitor, namely mepazine has been shown to have significant protective effects, either early, before the onset of acute disease or late, at the peak of disease in a study made by Me Guire et al. In the same study, it has also been determined that mepazine treatment does not affect peripheral regulatory T cells whereas from the previous studies it is known that the complete absence of MALT1 activity in knockout mice leads to a severe defect in the development of Foxp3+ Tregs which is consistent with the essential role of agonist-induced TCR signaling in the expression of Foxp3 and the development of Tregs. These findings further underline the role of Treg cells and the maintenance of immune homeostasis by maintaining the number of Tregs which further demonstrates that a balanced inhibition of TCR signaling which do not affect the development of Tregs is a promising for the treatment of autoimmune disease.

[0412] Further identified MALT1 inhibitors include compounds MLT-943 and MLT-985 having good bioavailability in rodents, however, MLT-943 treatment, in both rats and dogs, caused a rapid and dosedependent decrease in the number of peripheral Treg cells, resulting in disrupted immune homeostasis and an IPEX (immunodeficiency, poly-endocrinopathy, and enteropathy-X-linked)-like immune pathology after several weeks of treatment. Although these effects of the compound was reversible with the withdrawal of the same, it posed concerns about the long term use of MALT1 inhibitors. Furthermore, the pharmacodynamics of MLT-943 clearly indicate that a decrease of Treg cells is an on-target effect of the compound, which is also in agreement with the developmental and functional loss of Treg cells in MALT1 paracaspase mutant mice. However, such a strong reduction of Treg cells leading to an autoimmune phenotype was not observed following the efficient genetic inactivation of the MALT1 protease in adult mice or treatment with the less potent MALT1 inhibitor mepazine as mentioned above.

[0413] Therefore, when treating MS or symptoms of MS, the potency of the compound and its target play a role in the progress and / or the treatment of the disease and further side effects observed. Thus, MALT1 inhibitor having the highest potency does not always promise the best treatment outcome.

[0414] Surprisingly, the inventors have identified gliptins as MALT1 inhibitors. Gliptins are shown to bind to the allosteric site of MALT1 and therefore act intracellularly.

[0415] In line with the above, in accordance with the invention, the gliptin is administered to inhibit MALT1 , in particular the gliptin inhibits MALT1 allosterically, and / or the disease can be treated by the inhibition of MALT1 .

[0416] In a preferred embodiment, the to be treated subject has an altered, impaired and / or overexpressed level of MALT1.

[0417] Gliptins

[0418] Gliptins are a class of pharmaceutical compounds which are the inhibitors of dipeptidyl peptidase 4, also known as DPP-4 inhibitors. Now, the inventors identify a new target for the gliptins which is MALT 1 , an intracellular signalling protein that is widely expressed and known to be implicated in innate [natural killer cells (NK), dendritic cells (DC), and mast cells] and adaptive immune cells (T cells and B cells), where it signals proinflammatory gene expression downstream of several cell-surface receptors.Gliptins are known to bind the extracellular domain of DPP4, also known as CD26, which is a surface T cell activation antigen and has been shown to have DPP4 enzymatic activity, cleaving-off amino-terminal dipeptides with either L-proline or L-alanine at the penultimate position. Therefore, it plays a major role in glucose metabolism by N-terminal truncation and inactivation of the incretins glucagon-like peptide-1 (GLP) and gastric inhibitory protein (GIP). However, a large number of studies demonstrate clearly that CD26 / DPP4 also plays an integral role in the immune system, particularly in T cell activation.

[0419] Interestingly, a study conducted in a cohort of MS patients with different clinical forms revealed that sDPP4 concentration and DPP activity are significantly lower in MS patients than controls (Tejera-Alhambra et al.). This indicates that although DPP-4 are known to be implied in T cell activation, apparently the pathology of MS displays a different DPP-4 expression profile which is considered low based on the episode and / or form of the disease. Now, unexpectedly, the inventors have identified gliptins as a MALT1 inhibitor, particularly an allosteric MALT1 inhibitor which can act intracellularly and inhibit the activity of MALT1 as desired.

[0420] In line with the above, surprisingly, the inventors now have found that gliptins can bind to an allosteric site of the MALT1 and inhibit the same allosterically. As shown in the below examples, survival of Jurkat cells mimicking cytokine responses of Th1 and Th17 cells as in MS being induced by anti-CD3 and anti-CD28 antibodies in combination with recombinant human IL-17 and IFN-y are inhibited by a gliptin according to the invention.

[0421] Furthermore, example 2 below shows that gliptins does not inhibit CYP2D6 and CYP3A4 enzymatic activity contrary to mepazine.

[0422] In summary:

[0423] An aspect of the disclosure relates to a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating a MALT-1 mediated disease selected from an autoimmune disease, preferably multiple sclerosis, in a subject, preferably a human subject, in need thereof.

[0424] An embodiment is the pharmaceutical composition for use, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin or cetagliptin, preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, more preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin or alogliptin, most preferably wherein the gliptin is sitagliptin.

[0425] An embodiment is the pharmaceutical composition for use, wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin,gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, or cetagliptin, preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin or alogliptin.

[0426] An embodiment is the pharmaceutical composition for use, wherein the gliptin is not sitagliptin. An embodiment is the pharmaceutical composition for use, wherein the gliptin is an inhibitor of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1), preferably an allosteric inhibitor of MALT1.

[0427] An embodiment is the pharmaceutical composition for use, wherein the therapeutic effect of the gliptin comprises and / or results from inhibition of MALT1 , preferably wherein the inhibition of MALT1 is an allosteric inhibition resulting from binding of the gliptin to an allosteric site within MALT1.

[0428] An embodiment is the pharmaceutical composition for use, wherein the method comprises oral administration of the pharmaceutical composition.

[0429] An embodiment is the pharmaceutical composition for use, wherein the method results in inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside aberrant cells, preferably aberrant Th1 cells and / or Th17 cells.

[0430] An embodiment is the pharmaceutical composition for use, wherein the MS is clinically isolated syndrome (CIS), relapsing-remitting MS (RRMS), secondary progressive MS (SPMS), primary progressive MS (PPMS) or progressive relapsing MS (PRMS).

[0431] An embodiment is the pharmaceutical composition for use, wherein the MS is at pre-symptomatic stage, clinically isolated syndrome stage, relapsing-remitting stage, or secondary progressive stage.

[0432] An embodiment is the pharmaceutical composition for use, wherein the MALT-1 mediated disease selected from an autoimmune disease, preferably multiple sclerosis, is mediated by or dependent on MALT1 activity.

[0433] An embodiment is the pharmaceutical composition for use, wherein the method comprises the step of diagnosing a subject as suffering from the MALT-1 mediated disease selected from an autoimmune disease, preferably multiple sclerosis, or being at risk of suffering from the MALT-1 mediated disease selected from an autoimmune disease, preferably multiple sclerosis, mediated by or dependent on MALT1 activity.

[0434] An embodiment is the pharmaceutical composition for use, wherein the treatment is followed by radiotherapy and / or chemotherapy.

[0435] An embodiment is the pharmaceutical composition for use, wherein MALT1 expression status, and optionally DPP-4 expression status, comprises RNA expression analysis and / or protein expression analysis, preferably by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and / or immunoassay.

[0436] An embodiment is the pharmaceutical composition for use, wherein the method comprises the oral administration of the gliptin, pharmaceutically acceptable salt thereof or hydrate thereof, at a dose of 0.01 mg to 1000 mg per day.

[0437] An embodiment is the pharmaceutical composition for use, wherein the pharmaceutical composition is provided in an oral unit dosage form, preferably in the form of a tablet, comprising thegliptin, the pharmaceutically acceptable salt thereof or hydrate thereof, in an amount of 0.01 mg to 1000 mg.

[0438] An aspect of the disclosure relates to a therapeutic or prophylactic method of treating a MALT-1 mediated disease selected from multiple sclerosis, in a subject, preferably human subject, in need thereof, said method comprising the step of administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0439] An aspect of the disclosure relates to a use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of a MALT-1 mediated disease selected from multiple sclerosis, in a subject, preferably human subject, in need thereof.

[0440] NEUROLOGICAL DISORDERS, IN PARTICULAR GLIOMA

[0441] Reference will now be made in detail to general aspects and specific embodiments of the disclosed herein concepts, which unless specified otherwise, are to be assumed to be interrelated with each other and suitable to work in co-operation. While specific embodiments will be described, it will be understood that they are not intended to be read in isolation from other embodiments. On the contrary, the presented herein disclosure is intended to address and / or cover all alternatives, modifications, and equivalents, which may be included within the scope of the appended claims. One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice and should be construed as falling within the scope of the appended claims.

[0442] The general concept underlying the present disclosure originates from an unexpected finding of the inventors that gliptins, which are known to be DPP-4 inhibitors, can bind to an allosteric site on MALT1 and inhibit the same, advantageously reversibly.

[0443] Furthermore, but without wishing to be bound to the theory, it surprisingly appears that although gliptins were developed for targeting a disease target located extracellularly (catalytic domain of DPP-4) they seem to be inhibiting and allosterically binding to MALT1 (at least partially) intracellularly.

[0444] These surprising and unexpected findings lead the inventors to the presented herein novel use of the gliptins for the treatment of MALT1 -positive neurological cancers, in which subpopulation of neurological cancers, the benefits of such treatment are expected to greatly outweigh possible DPP-4 inhibition-related side-effects (if DPP-4 is still not silenced in such cancers), in particular in neurological cancer subpopulations wherein MALT1 is determined as overexpressed and / or DPP-4 expression is determined as corresponding to the baseline DPP-4 level or even lower, or advantageously, is even determined as lost (below detection level, or confirmed to be lost by e.g. genomic analysis).

[0445] A possible advantage of the presented herein approaches is that the currently known canonical MALT1 inhibitors display certain immunity-related side effects, which are either not known for one or more of gliptins, in particular the already approved gliptins. More details about MALT1 and its inhibition is explained below.MALT1

[0446] The Mucosa-associated lymphoid tissue lymphoma translocation 1 (MALT1) is a paracaspase and a central signalling protein which forms part of the intracellular CBM (CARMA / CARD-BCL10-MALT1) signalling complex downstream of receptors containing immunoreceptor tyrosine-based activation motif (ITAMs) as well as a subset of G-protein-coupled receptors (GPCRs) and receptor tyrosine kinases found in both hematopoietic and non-hematopoietic cell types. Following receptor activation, the CBM complex activates downstream signaling events via the scaffolding and protease functions of MALT1 that regulates key cellular processes, including lymphocyte development and activation of cells of the innate and adaptive immune system. Following TCR or BCR triggering, MALT 1 is required for activation of the canonical NF-kB pathway MALT1 partners with BCL10 and CARMA1 to form the CARMA1-BCL10-MALT1 signalosome complex (CBM), where MALT1 has been thought to act as a major protein scaffold that recruits downstream effector proteins to activate NF-kB signaling. MALT1 also plays a role in the activation of the non-canonical NF-kB pathway upon B cell activating factor receptor (BAFF-R) stimulation, a pathway that is needed for survival of marginal zone (MZ) B cells.

[0447] Thus, MALT1 has dual role: i) scaffold activity for the assembly of CBM complex and ii) proteolytic activity which is thought to promote NF-KB activation through the cleavage of the deubiquitinating enzyme A20 and of the NF-KB subunit RelB, CYLD and HOIL-1. Moreover, MALT1 possesses the capacity of autocleavage, which is an additional necessary step in NF-KB activation. MALT1 -dependent RelB cleavage results in its proteasomal degradation, thereby lowering total levels of RelB in activated B- and T cells. Through cleavage of RelB and A20, MALT1 thus expands the amplitude and duration of the NF-KB response in an IKK-independent manner. Interestingly, MALT1 also promotes NF-KB activation by its autoprocessing after Arg 149. The exact consequences of MALT1 autoprocessing remain unclear, but include an effect on NF-KB target gene expression downstream of nuclear NF-KB accumulation.

[0448] The role of the paracaspase activity of MALT1 has been studied in vivo using genetically modified knock-in (MALT1-PD) mice expressing an inactive form of the MALT1 protease.

[0449] MALT1 inhibitors

[0450] MALT 1 ’s enzymatic activity has stimulated efforts to develop specific MALT 1 inhibitors for research and therapeutic purposes. The first MALT1 inhibitor identified was z-VRPR-fmk, a modified tetrapeptide based on the optimal substrate Val-Arg-Pro-Arg of the Arabidopsis thaliana metacaspase AtmC9, conjugated to fluoromethyl ketone (fmk). This modified peptide irreversibly blocked MALT1 protease activity in an in vitro cleavage assay using recombinant MALT1 in a dose-dependent manner. It furthermore efficiently inhibited T cell activation and IL-2 secretion in JurkatT cells and in human antigen specific CTLs. Nagel and colleagues have identified three phenothiazine derivatives (mepazine, thioridazine, and promazine) as highly specific, noncompetitive and reversible MALT1 inhibitors. A concurrent study by Fontan and colleagues has identified the compound MI-2 as a selective MALT1 inhibitor. In contrast to phenothiazine derivatives, MI-2 is a covalent and irreversible inhibitor that acts on the catalytic centre of MALT1.It has been shown that reversible and irreversible inhibitors of MALT1 can act differently on autoimmune cells. The therapeutic potential of targeting MALT1 , although promising, has been hindered by concerns regarding the impact of MALT inhibition on Treg homeostasis.

[0451] MALT1 as a druqqable target for treating cancers

[0452] MALT1 protease function is silent in resting cells, including resting lymphocytes. In healthy lymphocytes its protease activity only becomes induced after antigen receptor ligation in T and B cells. However, MALT1 protease is constitutively active and drives growth and survival of BCR-addicted ABC DLBCL or apoptosis resistance in the context of the oncogenic API2-MALT1 fusion, identifying MALT1 as a target for lymphoma therapy (Rebeaud et al., 2008). MALT1 paracaspase is an attractive therapeutic target in BCR-addicted aggressive lymphomas, because within the CBM complex it acts downstream of most druggable upstream protein kinases including BTK, as well as all main oncogenic driver mutations in the BCR signaling pathway (Ngo et al., 2006). Importantly, and in contrast to the double-edge sword situation for DPP-4, increased MALT1 expression and / or activation is believed to contribute to tumorigenesis in a variety of different non-hematologic solid cancers including glioblastoma, breast cancer, osteosarcoma, pancreatic cancer, melanoma, lung carcinoma and prostate cancer (Jacobs et al., 2020; Ekambaram et al., 2018; Israel et al., 2021 ; Konczalla et al., 2019; Liu et al., 2020; McAuley et al., 99; Tan et al., 2021 ; Wang et al., 2017). In many cases, involvement of the alternative CARD10-containing CBM complex has been suggested.

[0453] MALT1 as a druqqable target for treating glioma

[0454] High MALT1 expression correlates with fast progression and short survival in glioblastoma multiforme (GBM), a yet incurable brain tumor (Jacobs et al., 2020; Fang et al, 2017). MALT1 was suggested to act downstream of EGFR and to induce mesenchymal transition in human glioblastoma-derived cell lines, thereby promoting tumor cell proliferation, survival, invasion and migration (Liu et al., 2020). Intriguingly, MALT1 protease activity is also required for expansion and viability of glioblastoma stemlike cells (GSC) in vitro and in vivo, which build up the reservoir of cancer cells and thus are thought to be a major cause for therapy resistance. Phenothiazine-derivatives like mepazine cause a marked decline in the expression of sternness markers in patient-derived GSC. Indeed, the reduced capacity of self-renewal relied on binding of mepazine to the allosteric site in MALT1 , presenting strong evidence that the effects are MALT1 protease-dependent (Jacobs et al., 2020). Lysosomal homeostasis has been shown to be critical for GBM cell survival (Le Joncour et al., 2019; Shingu et al., 2017). Mechanistically, MALT1 protease inhibition affects lysosomal homeostasis in GSC and thereby induces autophagy- and mTOR-driven cell death (Jacobs et al., 2020). Of note, even though other MALT1 inhibitors are more potent, phenothiazines like mepazine are able to cross the blood brain barrier, which may facilitate target occupancy and therapeutic use of this class of compounds in brain tumors.

[0455] Taken together, MALT1 protease controls survival, expansion and metastasis in a variety of non-hematologic solid cancers, revealing options for precision targeting of MALT1 beyond lymphomas.

[0456] MALT1 phenothiazines inhibitors and their undesired side-effectsIt is known that phenothiazines inhibit MALT1 Activity and IL-2 Induction in T Cells. Derivatives of medicinal active phenothiazines, including mepazine, have been identified as small molecule inhibitors of the MALT1 protease (Nagel 2012). These phenothiazines selectively inhibit cleavage activity of recombinant and cellular MALT1 by a noncompetitive mechanism. Mepazine has been evaluated as an antipsychotic and tranquilizing drug under the brand name Pacatai in the late 1950s and early 1960s. Whereas some clinical investigations provided evidence for antipsychotic effects, other studies failed to do so (Sarwer-Foner and Koranyi, 1957; Whittier et al., 1960). Some additional effects were reported after mepazine treatment, including a reduction of asthma attacks, indicating potential immunosuppressive activity (Sarwer-Foner and Koranyi, 1957). While the phenothiazine-derivatives, mepazine and (S)-mepazine, are the first known allosteric MALT1 inhibitors with medium potency, the compounds are not highly selective for MALT1 and antagonize other proteins such as G protein coupled receptors (GPCR). Thus, mepazine may affect cellular functions independent of MALT 1. For instance, mepazine impairs CYP450 isoenzymes which are the basic enzymes involved in Phase I biotransformation. The most important role in biotransformation belongs to CYP3A4, CYP2D6, CYP2C9, CYP2C19 and CYP1A2. Inhibition and induction of CYP isoenzymes caused by drugs are important and clinically relevant pharmacokinetic mechanisms of drug interaction and for assessing risks of drugs. Mepazine was shown to strongly interfere with CYP450 activity causing its almost full inhibition, likely through interference due to CYP2D6 inhibition. Furthermore, mepazine is almost completely silencing CYP450 metabolic activity also likely due to interfering with CYP2D6. Moreover, despite its effects on the MALT1 activity, this compound shows some side effects like tardive dyskinesia which limits its clinical study (Liang et al. 2021).

[0457] General aspects of the present disclosure

[0458] In view of the above facts, the realization that gliptins inhibit MALT1 opens promising options for aiding the treatment of neurological cancers expressing MALT1 with the potential advantage of avoiding the side effects of the currently known MALT1 inhibitors.

[0459] In view of the above, in a first general aspect, the disclosure relates to a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic method of treating neurological cancer in a human subject in need of the treatment, wherein the neurological cancer is positive for expression of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1).

[0460] In another aspect, the invention provides a therapeutic method of treating neurological cancer (preferably brain cancer) in a human subject in need of the treatment, wherein the cancer is positive for expression of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1), said method comprising the step of administering to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof; and optionally further comprising a diagnostic step preceding the step of administering, wherein the diagnostic step comprises determining MALT1 status, and optionally also DPP-4 status, in a sample obtained from the human subject.In yet a further aspect, the invention provides the use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of neurological cancer (preferably brain cancer) in a human subject in need of the treatment, wherein the cancer is positive for expression of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1).

[0461] In a further aspect, pharmaceutical composition forthe disclosed herein use is provided, wherein the therapeutic effect of the gliptin comprises and / or results from inhibition of MALT 1 , preferably wherein the inhibition of MALT1 is an allosteric inhibition resulting from binding of the gliptin to an allosteric site within MALT1.

[0462] In an embodiment, pharmaceutical composition forthe disclosed herein use is provided, wherein the gliptin is an inhibitor of MALT 1 , preferably an allosteric inhibitor of MALT 1.

[0463] In an embodiment, pharmaceutical composition forthe disclosed herein use is provided, wherein the method results in inhibition of intracellular MALT1 activity.

[0464] In another aspect, pharmaceutical composition forthe disclosed herein use is provided, wherein the cancer is negative for expression of dipeptidyl peptidase-4 (DPP-4) or comprises DPP-4 expression considered as unchanged (corresponding to baseline expression) or lower as compared to a healthy reference tissue, preferably being the tissue from which the cancer originated and / or non-cancerous tissue surrounding the cells of the cancer, advantageously being a healthy tissue of the subject.

[0465] In an embodiment, pharmaceutical composition forthe disclosed herein use is provided, wherein MALT1 expression in the cancer is assessed as MALT1 overexpression; e.g. as compared to a reference baseline expression expected for a healthy reference tissue or healthy cell type such as glial cell type.

[0466] In an embodiment, pharmaceutical composition forthe disclosed herein use is provided, wherein the cancer is mediated by / driven by or dependent on MALT1 activity.

[0467] Gliptins

[0468] Gliptins are a class of pharmaceutical compounds which are the inhibitors of dipeptidyl peptidase 4, and therefore are also known as DPP-4 inhibitors. Gliptins are known to bind the extracellular domain of DPP4, also known as CD26, which is a surface T cell activation antigen and has been shown to have DPP4 enzymatic activity, cleaving-off amino-terminal dipeptides with either L-proline or L-alanine at the penultimate position. Therefore, it plays a major role in glucose metabolism by N-terminal truncation and inactivation of the incretins glucagon-like peptide-1 (GLP) and gastric inhibitory protein (GIP). However, a large number of studies demonstrate clearly that CD26 / DPP4 also plays an integral role in the immune system, particularly in T cell activation. Examples of known gliptins with verified DPP-4 inhibition activity include Sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin, Alogliptin Teneligliptin, Trelagliptin, Omarigliptin, Evogliptin, Gosogliptin, Retagliptin Dutogliptin, Neogliptin, Denagliptin, Cofrogliptin, Fotagliptin, Prusogliptin, Cetagliptin. As already explained above, the inventor has surprisingly realized and that gliptins can alco inhibit MALT1. Furthermore, in silico modelling of gliptin docking to MALT1 indicates that gliptins show an interaction with allosteric MALT1 -target site, see Example 3.In an embodiment, the disclosure relates to a pharmaceutical composition, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin or cetagliptin, preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, more preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin or alogliptin, most preferably wherein the gliptin is sitagliptin

[0469] In an embodiment, pharmaceutical composition forthe disclosed herein use is provided, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin;

[0470] In an embodiment, the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin

[0471] In a next embodiment, the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin, and alogliptin. In yet another embodiment, the gliptin is sitagliptin.

[0472] In an alternative embodiment, pharmaceutical composition for the disclosed herein use is provided, wherein the gliptin is not sitagliptin.

[0473] In a possible embodiment, pharmaceutical composition forthe disclosed herein use is provided, wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, and cetagliptin.

[0474] In an embodiment, the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin.

[0475] In yet another embodiment, the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin.

[0476] In an embodiment, the gliptin is linagliptin, vildagliptin, saxagliptin and alogliptin.

[0477] In an embodiment, the gliptin is linagliptin.

[0478] Exemplary structures of gliptins are shown below:

[0479]

[0480]

[0481]

[0482]

[0483] In one embodiment, the gliptin is sitagliptin.

[0484] In another embodiment, the gliptin is vildagliptin.

[0485] In yet another embodiment, the gliptin is saxagliptin.

[0486] In a further embodiment, the gliptin is linagliptin.

[0487] In another embodiment, the gliptin is gemigliptin.

[0488] In yet another embodiment, the gliptin is anagliptin.

[0489] In a further embodiment, the gliptin is alogliptin.

[0490] In one embodiment, the gliptin is retagliptin.

[0491] In another embodiment, the gliptin is evogliptin.

[0492] In yet another embodiment, the gliptin is trelagliptin.

[0493] In a further embodiment, the gliptin is teneligliptin.

[0494] In another embodiment, the gliptin is gosogliptin.

[0495] In yet another embodiment, the gliptin is omarigliptin.

[0496] Sitagliptin

[0497] Sitagliptin (C16H15F6N5O; (3R)-3-amino-1-[3-(trifluoromethyl)-6,8-dihydro-5H-[1 ,2,4]triazolo[4,3-a]pyrazin-7-yl]-4-(2,4,5-trifluorophenyl)butan-1-one) is the first antidiabetic agent from the class of dipeptidyl peptidase-4 (DPP-4) enzyme inhibitors. It increases the amount of circulating incretins, which stimulate insulin secretion and inhibit glucose production (Choy et al, 2007).

[0498] DPP-4 is an enzyme that breaks down incretin hormones such as glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide-1 (GIP) (Plosker, 2014). By inhibiting DPP-4, sitagliptin results in improved glucose control. Additionally, sitagliptin decreases the rate at which glucose is produced by the liver. This decreased hepatic glucose production contributes further to improved glycaemic control in patients with type- 2 diabetes (Katsuno et al, 2013; Sakura et al, 2016).

[0499] Several preclinical studies have also shown that DPP-4 inhibitors can improve endothelial function, reduce oxidative stress, and attenuate inflammation, all of which are key factors in the pathogenesis of cardiovascular diseases (Hanssen and Jandeleit-Dahm, 2019; Liu et al, 2019; Chikata et al, 2022). Clinical trials have also demonstrated that sitagliptin can improve left ventricular function and reduce circulating biomarkers of inflammation and oxidative stress in patients with heart failure and preserved ejection fraction (Zannad et al, 2015). The underlying mechanism for these effects is related to the inhibition of DPP-4, which leads to increased levels of GLP-1 and other vasoactive peptides that enhance endothelial function and reduce inflammation (Kim et al, 2014).

[0500] Administration

[0501] An embodiment is the pharmaceutical composition for use according to the invention, wherein the method comprises oral administration of the pharmaceutical composition.

[0502] Dose regimenIn embodiments, is the dose of gliptin, preferably the once daily dose of the gliptin is preferably less than 1000 mg, preferable less than 500 mg, less than 250 mg, less than 200 mg, less than 150 mg, or equal to or less than 100 mg, even more preferably, equal to or less than 50 mg, 25 mg, 20 mg, 12,5 mg, 12 mg, 10 mg, 6,25 mg, 5 mg or 2,5 mg. For some gliptins, preferred (once daily) doses are listed in Table B here below.

[0503] TABLE B. Examples of doses for a series of gliptins for use according to the invention

[0504]

[0505] Neurological cancers including brain cancers

[0506] All types of brain cancers may produce symptoms that vary depending on the part of the brain involved. These may include headaches, seizures, problem with vision, vomiting, and mental changes. The headache is classically worse in the morning and goes away with vomiting. More specific problems may include difficulty in walking, speaking and with sensation. As the disease progresses unconsciousness may occur.

[0507] The underlying cause of most brain tumors is unknown. Risk factors that may occasionally be involved include a number of inherited conditions, such as neurofibromatosis, as well as exposure to the external factors such as radiation, chemicals, or viruses; such as ionizing radiation, vinyl chloride, or the Epstein-Barr virus. The most common types of primary tumors in adults are: meningiomas (usually benign), and astrocytomas such as glioblastomas. In children, the most common type is a malignant medulloblastoma, while a rarer type that is believed to start during embryonic development is neuroblastoma. Diagnosis is usually by medical examination along with computed tomography or magnetic resonance imaging. This is then often confirmed by a biopsy. Based on the findings, the tumors are divided into different grades of severity.

[0508] Treatment may include different combinations of surgery, radiation therapy and chemotherapy. Anticonvulsant medication may be needed if seizures occur. Dexamethasone and furosemide may beused to decrease swelling around the tumor. Some tumors grow gradually, requiring only monitoring and possibly needing no further intervention. Treatments that use a person's immune system are being studied. Outcome varies considerably depending on the type of tumor and how far it has spread at diagnosis. Glioblastomas usually have poor outcomes while meningiomas usually have good outcomes. The average five-year survival rate for brain cancer in the United States is 33%.

[0509] As most of the majority of known neurological cancers are brain cancers, in the context of the present disclosure, it can be assumed that particular embodiments described for brain cancer treatment can equally be applied to analogous neurological cancers present in the spine or localized elsewhere in the body as a likely result of metastasis of the primary cancer in the brain or, less often, in the spinal cord. In the context of such metastases, as used herein, it should be construed that any tumor mass that is present outside of the subject’s central nervous system (CNS), usually being skull area, but appears to be a result of metastasis from a primary CNS / brain cancer or patho-physiologically appears to have such origin, is to be considered as falling within the scope of the terms “neurological caner” and / or “brain cancer” as used herein in the specific contexts, for the purpose of the disclosed herein treatments, unless specified otherwise.

[0510] In advantageous possible embodiments, pharmaceutical composition for the disclosed herein uses is provided, wherein the neurological cancer is brain cancer and / or is glioma, neuroblastoma, or medulloblastoma; preferably being glioma or neuroblastoma, more preferably being glioblastoma or gliosarcoma, even more preferably being malignant glioblastoma, most preferably being glioblastoma multiforme.

[0511] In an embodiment, the neurological cancer is brain cancer, preferably classified as glioma, neuroblastoma, or medulloblastoma.

[0512] In an embodiment, the cancer is glioma or neuroblastoma.

[0513] In a specific embodiment, the cancer is glioma, preferably being glioblastoma or gliosarcoma, In a particular embodiment, the cancer is malignant glioblastoma, most preferably being glioblastoma multiforme.

[0514] In an advantageous embodiment, the treatment comprises administering to the human subject of an effective amount of the compound, preferably directly into the cancer or into the cavity remaining after surgical removal of the cancer, possibly in combination with one or more chemotherapeutic agent and / or anticancer agent specific to the cancer of the subject.

[0515] In an embodiment, the treatment is followed by radiotherapy and / or chemotherapy.

[0516] In an embodiment, MALT1 expression status, and optionally DPP-4 expression status, comprises RNA expression analysis and / or protein expression analysis, preferably by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and / or immunoassay.

[0517] In another general aspect, the invention also relates to an in vitro method for determining an increased probability that a neurological cancer in a human subject will respond to the treatment with a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, wherein advantageously the gliptin is selected from wherein the gliptin is selected from any one or more of sitagliptin, vildagliptin, saxagliptin,linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin, and cetagliptin; the method comprising the steps of:

[0518] - providing a test first sample obtained from the subject;

[0519] - detecting the presence of MALT1 expression and preferably assessing the expression level of MALT 1 in the first test sample;

[0520] - preferably, comparing the assessed expression level of MALT1 to a reference MALT1 expression level;

[0521] wherein finding that MALT1 expression is present in the first test sample and, preferably that the expression level of MALT1 assessed in the first test sample is higher as compared to the reference MALT1 expression level is indicative of an increased probability that the subject will respond to the treatment.

[0522] In an advantageous embodiment, an in vitro method of the disclosure is provided, comprising the steps of:

[0523] - providing a second test sample obtained from the subject;

[0524] - detecting the presence of DPP-4 expression or optionally assessing the expression level of DPP-4 or DPP-4 enzymatic activity level in the second test sample;

[0525] - preferably, comparing the assessed expression level or enzymatic activity level of DPP-4 to a reference DPP-4 expression level or a reference DPP-4 enzymatic activity level, respectively;

[0526] wherein finding that DPP-4 expression is absent in the second test sample or optionally that the expression level of DPP-4 or DPP-4 enzymatic activity level assessed in the second test sample is lower as compared to the reference DPP-4 expression level or a reference DPP-4 enzymatic activity level, respectively; preferably finding that DPP-4 expression is absent in the second test sample is indicative of an increased probability that the subject will respond to the treatment, wherein the response is further associated with an increased probability of not causing adverse DPP-4-inhibition-associated side effects.

[0527] In an embodiment of the disclosed herein in vitro method, the first test sample and / or the second test sample is or comprises biopsy sample, cerebrospinal fluid (CSF) sample, or a blood sample, preferably being biopsy sample or the cerebrospinal fluid (CSF) sample, even more preferably being biopsy sample.

[0528] In an embodiment of the disclosed herein in vitro method, detecting the presence of expression and / or assessing the expression level comprises RNA analysis, preferably comprising RNA-sequencing or reverse transcription-quantitative polymerase chain reaction (RT-qPCR), more preferably comprising RT-qPCR.

[0529] In an embodiment of the disclosed herein in vitro method, detecting the presence of expression and / or assessing the expression level comprises protein analysis, preferably comprising an immunoassay, more preferably comprising ELISA.

[0530] In an advantageous embodiment of the disclosed herein in vitro method, the first test sample and the second test sample are the same test sample, preferably wherein detecting the presence of MALT1 and DPP-4 expression, optionally assessing the expression level of MALT1 and DPP-4 isperformed as part of the same assay, more preferably being a multiplex RT-qPCR assay or an immunoassay.

[0531] In an embodiment of the disclosed herein in vitro method, the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin; preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin; more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin, and alogliptin, most preferably wherein the gliptin is sitagliptin;

[0532] In an embodiment of the disclosed herein in vitro method, the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, and cetagliptin; preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin; more preferably wherein the gliptin is of vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin; even more preferably of linagliptin, vildagliptin, saxagliptin and alogliptin, most preferably wherein the gliptin is linagliptin.

[0533] In a further embodiment of the disclosed herein in vitro method, the reference MALT1 expression value, and optionally the reference DPP-4 expression value, is defined and has (have) been predetermined based on assessing MALT1 expression levels, and optionally DPP-4 expression levels, respectively, in a plurality of control samples obtained from healthy control subjects.

[0534] In an embodiment of the disclosed herein in vitro method, the reference MALT1 expression value, and optionally the reference DPP-4 expression value, is determined based on assessing normal MALT1 expression levels, and optionally normal DPP-4 expression levels, respectively, in a healthy tissue sample obtained from the subject, preferably being a healthy tissue corresponding to the tissue from which the cancer originated, optionally being healthy non-cancerous tissue surrounding the cancer cells.

[0535] In an embodiment of the disclosed herein in vitro method, the expression level of MALT1 assessed for the first test sample is at least 2 fold higher than the reference MALT1 expression value, preferably being at least 5 fold higher; more preferably being at least 10 fold higher.

[0536] In summary:

[0537] An aspect of the disclosure relates to a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic method of treating neurological cancer in a human subject in need of the treatment, wherein the neurological cancer is positive for expression of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1)

[0538] An embodiment is the pharmaceutical composition for use, wherein the gliptin is an inhibitor of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1), preferably an allosteric inhibitor of MALT1.An embodiment is the pharmaceutical composition for use, wherein the therapeutic effect of the gliptin comprises and / or results from inhibition of MALT1 , preferably wherein the inhibition of MALT1 is an allosteric inhibition resulting from binding of the gliptin to an allosteric site within MALT1.

[0539] An embodiment is the pharmaceutical composition for use, wherein the neurological cancer is negative for expression of dipeptidyl peptidase-4 (DPP-4) or comprises DPP-4 expression considered as unchanged to or lower as compared to a healthy reference tissue, preferably being the tissue from which the cancer originated and / or non-cancerous tissue surrounding the cells of the cancer, advantageously being a healthy tissue of the subject.

[0540] An embodiment is the pharmaceutical composition for use, wherein MALT1 expression in the neurological cancer is assessed as MALT1 overexpression.

[0541] An embodiment is the pharmaceutical composition for use, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin; preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin; more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin, and alogliptin, most preferably wherein the gliptin is sitagliptin.

[0542] An embodiment is the pharmaceutical composition for use, wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, and cetagliptin; preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin; more preferably wherein the gliptin is of vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin; even more preferably of linagliptin, vildagliptin, saxagliptin and alogliptin, most preferably wherein the gliptin is linagliptin.

[0543] An embodiment is the pharmaceutical composition for use, wherein the method comprises oral administration of the pharmaceutical composition.

[0544] An embodiment is the pharmaceutical composition for use, wherein the MALT-1 mediated disease selected from a neurological cancer such as glioma is mediated by or dependent on MALT1 activity.

[0545] An embodiment is the pharmaceutical composition for use, wherein the method comprises the step of diagnosing a subject as suffering from the MALT-1 mediated disease selected from a neurological cancer such as glioma, or being at risk of suffering from the MALT-1 mediated disease selected from a neurological cancer such as glioma, mediated by or dependent on MALT1 activity.

[0546] An embodiment is the pharmaceutical composition for use, wherein the neurological cancer is brain cancer and / or is glioma, neuroblastoma, or medulloblastoma; preferably being glioma or neuroblastoma, more preferably being glioblastoma or gliosarcoma, even more preferably being malignant glioblastoma, most preferably being glioblastoma multiforme.

[0547] An embodiment is the pharmaceutical composition for use, wherein the treatment comprises administering to the human subject of an effective amount of the compound, preferably directly into the neurological cancer or into the cavity remaining after surgical removal of the neurological cancer,possibly in combination with one or more chemotherapeutic agent and / or anticancer agent specific to the cancer of the subject.

[0548] An embodiment is the pharmaceutical composition for use, wherein the treatment is followed by radiotherapy and / or chemotherapy.

[0549] An embodiment is the pharmaceutical composition for use, wherein MALT1 expression status, and optionally DPP-4 expression status, comprises RNA expression analysis and / or protein expression analysis, preferably by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and / or immunoassay.

[0550] An embodiment is the pharmaceutical composition for use, wherein the method comprises the oral administration of the gliptin, pharmaceutically acceptable salt thereof or hydrate thereof, at a dose of 0.01 mg to 1000 mg per day.

[0551] An embodiment is the pharmaceutical composition for use, wherein the pharmaceutical composition is provided in an oral unit dosage form, preferably in the form of a tablet, comprising the gliptin, the pharmaceutically acceptable salt thereof or hydrate thereof, in an amount of 0.01 mg to 1000 mg.

[0552] An aspect of the disclosure relates to a therapeutic or prophylactic method of treating a MALT-1 mediated disease selected from glioma, in a subject, preferably human subject, in need thereof, said method comprising the step of administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0553] An aspect of the disclosure relates to a use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of a MALT-1 mediated disease selected from glioma, in a subject, preferably human subject, in need thereof.

[0554] An aspect of the disclosure relates to an in vitro method for determining an increased probability that a neurological cancer in a human subject will respond to the treatment with

[0555] a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, wherein the gliptin is selected from wherein the gliptin is selected from any one or more of sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, , cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin, and cetagliptin; the method comprising the steps of:

[0556] - providing a test first sample obtained from the subject;

[0557] - detecting the presence of MALT1 expression and preferably assessing the expression level of MALT 1 in the first test sample;

[0558] - preferably, comparing the assessed expression level of MALT1 to a reference MALT1 expression level;

[0559] wherein finding that MALT1 expression is present in the first test sample and preferably that the expression level of MALT1 assessed in the first test sample is higher as compared to the reference MALT 1 expression level is indicative of an increased probability that the subject will respond to the treatment.An embodiment is the in vitro method, further comprising the steps of:

[0560] - providing a second test sample obtained from the subject;

[0561] - detecting the presence of DPP-4 expression or optionally assessing the expression level of DPP-4 or DPP-4 enzymatic activity level in the second test sample;

[0562] - preferably, comparing the assessed expression level or enzymatic activity level of DPP-4 to a reference DPP-4 expression level or a reference DPP-4 enzymatic activity level, respectively; wherein finding that DPP-4 expression is absent in the second test sample or optionally that the expression level of DPP-4 or DPP-4 enzymatic activity level assessed in the second test sample is lower as compared to the reference DPP-4 expression level or a reference DPP-4 enzymatic activity level, respectively; preferably finding that DPP-4 expression is absent in the second test sample is indicative of an increased probability that the subject will respond to the treatment, wherein the response is further associated with an increased probability of not causing adverse DPP-4-inhibition-associated side effects.

[0563] An embodiment is the in vitro method, wherein the first test sample and / or the second test sample is or comprises biopsy sample, cerebrospinal fluid (CSF) sample, or a blood sample, preferably being biopsy sample or the cerebrospinal fluid (CSF) sample, even more preferably being biopsy sample.

[0564] An embodiment is the in vitro method, wherein detecting the presence of expression and / or assessing the expression level comprises RNA analysis, preferably comprising RNA-sequencing or reverse transcription-quantitative polymerase chain reaction (RT-qPCR), more preferably comprising RT-qPCR.

[0565] An embodiment is the in vitro method, wherein detecting the presence of expression and / or assessing the expression level comprises protein analysis, preferably comprising an immunoassay, more preferably comprising ELISA.

[0566] An embodiment is the in vitro method, wherein the first test sample and the second test sample are the same test sample, preferably wherein detecting the presence of MALT1 and DPP-4 expression, optionally assessing the expression level of MALT1 and DPP-4 is performed as part of the same assay, more preferably being a multiplex RT-qPCR assay or an immunoassay.

[0567] An embodiment is the in vitro method, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin;

[0568] preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin;

[0569] more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin, and alogliptin, most preferably wherein the gliptin is sitagliptin;

[0570] and / or

[0571] wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, and cetagliptin;preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin;

[0572] more preferably wherein the gliptin is of vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin;

[0573] even more preferably of linagliptin, vildagliptin, saxagliptin and alogliptin,

[0574] most preferably wherein the gliptin is linagliptin.

[0575] An embodiment is the in vitro method, wherein the reference MALT1 expression value, and optionally the reference DPP-4 expression value, is defined and has (have) been predetermined based on assessing MALT1 expression levels, and optionally DPP-4 expression levels, respectively, in a plurality of control samples obtained from healthy control subjects.

[0576] An embodiment is the in vitro method, wherein the reference MALT1 expression value, and optionally the reference DPP-4 expression value, is determined based on assessing normal MALT1 expression levels, and optionally normal DPP-4 expression levels, respectively, in a healthy tissue sample obtained from the subject, preferably being a healthy tissue corresponding to the tissue from which the cancer originated, optionally being healthy non-cancerous tissue surrounding the cancer cells.

[0577] An embodiment is the in vitro method, wherein the expression level of MALT1 assessed for the first test sample is at least 2 fold higher than the reference MALT1 expression value, preferably being at least 5 fold higher; more preferably being at least 10 fold higher.

[0578] MULTIPLE MYELOMA

[0579] The present invention is in the field of drug repurposing, wherein in an effort to address the unmet need for an improved treatment for multiple myeloma, the present inventors have identified gliptins for use in the treatment of multiple myeloma. In an unexpected finding, the inventors have now revealed that the gliptins, which are traditionally known to be DPP-4 inhibitors, can also bind to an allosteric site (pocket) on MALT1 and can act as an allosteric MALT1 inhibitor. This surprising and unexpected finding leads to a novel use of the gliptins for the treatment of multiple myeloma.

[0580] Multiple Myeloma (MM) & NF-KB signaling

[0581] Multiple myeloma is a hematological malignancy characterized by the uncontrolled proliferation of plasma cells in the bone marrow. The dysregulated progression of MM cells involve genetic, epigenetic molecular aberrations in vital genes, microenvironmental factors and signaling pathways. Certain microenvironmental factors, such as BM microenvironment, including insulin-like growth factor, proliferation-inducing ligand (APRIL), and B-cell activating factor (BAFF), wherein the B-cell maturation antigen (BCMA) is a critical transmembrane glycoprotein expressed on MM plasma cells and acts as a receptor for ligands such as BAFF and APRIL in the BM microenvironment.While the signaling pathways involve the NF-KB pathway, PI3K / Akt / mTOR signaling, and JAK / STAT signaling. Particularly, the NF-KB (nuclear factor kappa-light-chain-enhancer of activated B cells) signaling pathway plays a pivotal role in the pathogenesis and progression of multiple myeloma. NF-KB is a family of transcription factors that regulate the expression of genes involved in cell survival, proliferation, and resistance to apoptosis and immune responses.

[0582] In multiple myeloma, constitutive activation of NF-KB signaling occurs due to genetic mutations, aberrant protein expression, and interactions with the bone marrow microenvironment. One such pathway involve factors within the BM microenvironment, including insulin-like growth factor, proliferation-inducing ligand (APRIL), and B-cell activating factor (BAFF), which can directly or indirectly stimulate NF-KB activation in plasma cells and MM cells.

[0583] Consequently, targeting the NF-KB signaling pathway has been the current therapeutic strategy for the treatment of MM. For example: thalidomide analogues, lenalidomide and pomalidomide act by downregulating IRF4 and subsequent inhibition of B-cell receptor-dependent NF-KB activity; proteosome inhibitors, bortezomib, carfilzomib, ixazomib and marizomib act by inducing activation of canonical NF-KB signaling in MM; similarly Anti-PD-1 / PD-L1 monoclonal antibodies, pembrolizumab, atezolizumab, avelumab, durvalumab, nivolumab etc., mainly target NF-KB signaling through the inhibition of ligand-receptor interaction.

[0584] MALT1 mediated NF-KB signaling & Multiple Myeloma (MM)

[0585] In more recent studies, an alternative pathway of activation of NF-KB signaling involve mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1). MALT1 is known from innate (e.g., natural killer cells NK, dendritic cells DC, and mast cells) and adaptive immune cells (e.g., T cells and B cells), is the only human paracaspase which is ubiquitously expressed in several cell types. MALT1 is highly expressed in MM cells and is critical for cell growth and viability.

[0586] MALT1 functions as a pivotal player in the oncogenic process, in which its aberrant activation contributes to the dysregulation of cellular signaling pathways, including NF-KB signaling pathway. Specifically, MALT1 acts as an adaptor protein, forming a signaling complex known as CARD11-BCL10-MALT1 (CBM) in conjunction with caspase recruitment domain family member 11 (CARMA1 / CARD11) and BCL10. This complex, in turn, regulates coupled upstream signaling events of the canonical NF-KB via recruitment of E3 ubiquitin ligase, tumor necrosis factor receptor-associated factor 6. The dysregulated NF-KB signaling induced by MALT1 in MM promotes oncogenic processes by enhancing the production of anti-apoptotic proteins (e.g., Bcl-2, Bcl-xL), pro-inflammatory cytokines (e.g., IL-6, TNF-a), and angiogenic factors (e.g., VEGF), ultimately contributing to the development and progression of the disease (Yao et al. 2024).

[0587] This creates a microenvironment that supports tumor growth, enhances drug resistance, and promotes immune evasion. Furthermore, NF-KB activation in stromal cells within the bone marrow microenvironment fosters interactions that protect myeloma cells from therapeutic agents. Hence, sustained NF-KB activation in MM cells contributes to uncontrolled proliferation, enhanced survival mechanisms, and resistance to antimyeloma therapies.Consequently, targeting the NF-KB signaling pathway by blocking upstream activators, such as MALT1 , emerges as a promising therapeutic strategy to disrupt these critical cellular processes and combat MM effectively. This approach, particularly when combined with existing therapies, hold the potential to overcome drug resistance, suppress tumor growth, and improve patient outcomes in multiple myeloma.

[0588] Surprisingly, the inventors have identified gliptins as (intracellular acting) allosteric MALT1 inhibitors. Gliptins are shown to bind to the allosteric site (pocket) of MALT1 and therefore act intracellularly. This opens a whole new avenue in the treatment of multiple myeloma using gliptins. Gliptins as MALT1 inhibitors

[0589] Gliptins are a class of pharmaceutical compounds which are traditionally the inhibitors of dipeptidyl peptidase 4, also known as DPP-4 inhibitors. Gliptins are known to bind the extracellular domain of DPP4, also known as CD26, which is a surface T cell activation antigen and has been shown to have DPP4 enzymatic activity, cleaving-off amino-terminal dipeptides with either L-proline or L-alanine at the penultimate position.

[0590] However, the surprising finding of the present invention reveals that the gliptins show strong binding with MALT1 , preferably at an allosteric site (pocket) of MALT1. Hence, the present invention provides a method of treatment or prevention of multiple myeloma using gliptins (or a pharmaceutically acceptable salt thereof and hydrates thereof), preferably wherein multiple myeloma is mediated by either of: MALT1 overexpression or MALT1 mediated activation of NF-KB signaling pathway or induced levels of cytokines regulated by MALT1 -mediated NF-KB signaling pathway.

[0591] MALT1 inhibitors

[0592] MALT 1 ’s enzymatic activity has stimulated efforts to develop specific MALT 1 inhibitors for research and therapeutic purposes. The first MALT1 inhibitor identified was z-VRPR-fmk, a modified tetrapeptide based on the optimal substrate Val-Arg-Pro-Arg of the Arabidopsis thaliana metacaspase AtmC9, conjugated to fluoromethyl ketone (fmk). This modified peptide irreversibly blocked MALT1 protease activity in an in vitro cleavage assay using recombinant MALT1 in a dose-dependent manner. Further, three phenothiazine derivatives (mepazine, thioridazine, and promazine) as highly specific, noncompetitive and reversible MALT1 inhibitors were identified. A concurrent study identified MLT-748 and MI-2 like selective MALT1 inhibitor. In contrast to phenothiazine derivatives, MI-2 is a covalent and irreversible inhibitor that acts on the catalytic centre of MALT1.

[0593] It has been shown that reversible and irreversible inhibitors of MALT1 can act differently at cellular level. The therapeutic potential of targeting MALT1 , although promising, has been hindered by concerns regarding efficacy, potential side-effects and / or interference with CYP450 pathway.

[0594] For example, mepazine or (S)-mepazine, despite their good MALT1 inhibition, it is not highly selective for MALT1 and antagonize other proteins such as G protein coupled receptors (GPCR). Additionally, mepazine shows side effects like tardive dyskinesia which limits its clinical study (Liang et al. 2021a). Additionally, mepazine also interferes in the CYP450 and CYP2D4 pathway. Further identified MALT1 inhibitors include compounds MLT-943 and MLT-985 when tested in both rats and dogs, caused a disrupted immune homeostasis and an IPEX (immunodeficiency, poly-endocrinopathy, and enteropathy-X-linked)-like immune pathology.Mepazine has been evaluated as an antipsychotic and tranquilizing drug under the brand name Pacatai in the late 1950s and early 1960s. Whereas some clinical investigations provided evidence for antipsychotic effects, other studies failed to do so (Sarwer-Foner and Koranyi, 1957; Whittier et al., 1960). Some side effects were reported after mepazine treatment, including a reduction of asthma attacks, indicating potential immunosuppressive activity (Sarwer-Foner and Koranyi, 1957).

[0595] While the phenothiazine-derivatives, mepazine and (S)-mepazine, are the first known allosteric MALT1 inhibitors with medium potency, the compounds are not highly selective for MALT1 and antagonize other proteins such as G protein coupled receptors (GPCR). Thus, mepazine may affect cellular functions independent of MALT1 and, for instance, mepazine impairs CYP450 isoenzymes. Mepazine is doing bad on CYP450 activity (almost full inhibition) by interfering due to CYP2D6 inhibition and mepazine is almost completely silencing CYP450 metabolic activity due to interfering with CYP2D6. Moreover, despite the MALT1 activity, this compound shows some side effects like tardive dyskinesia which limits its clinical study (Liang et al. 2021a).

[0596] Overall, MALT1 has emerged as an exploratory therapeutic target in the recent times. Various MALT1 inhibitors have been under development and very few of them being investigated in the early clinical stages. For example, the few of the most advanced MALT1 inhibitor currently being investigated are MPT-0118, ABBV-525, CTX-177, safimaltib and SGR-1505, however none for the treatment of multiple myeloma. Out of all, Phase II is the highest development phase wherein MPT-0118 is being studied. Compared to the gliptins, it is too early for these MALT1 inhibitors to conclude on their safety profile. On the other hand, while the gliptins had been a well-established class of compounds, thoroughly studied for its safety and eventually accepted by the drug regulatory agencies of various countries including in USA, Europe, Japan, South Korea, Russia and China.

[0597] The same is eminent from the examples, wherein gliptins emerges over some of these MALT1 inhibitors in terms of either better or equal MALT1 inhibition, better or equal cytotoxicity and effectiveness against multiple myeloma and better safety profile.

[0598] In one aspect, the present disclosure relates to a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating multiple myeloma disease in a human subject in need thereof.

[0599] In another aspect, the present disclosure relates to a therapeutic or prophylactic method of treating multiple myeloma disease in a human subject in need thereof, said method comprising the administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0600] In yet another aspect, the present disclosure relates to use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of multiple myeloma disease in a human subject in need thereof.

[0601] In one embodiment, the gliptins are selected from one or more of: sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin,gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin or cetagliptin.

[0602] In a preferred embodiment, the gliptins are selected from one or more of: sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin.

[0603] In a more preferred embodiment, the gliptin are selected from one or more of: sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin.

[0604] In another more preferred embodiment, the gliptin are selected from one or more of: sitagliptin, linagliptin, vildagliptin, saxagliptin or alogliptin.

[0605] In yet another most preferred embodiment, the gliptin is sitagliptin.

[0606] In one embodiment, the gliptin is sitagliptin.

[0607] In another embodiment, the gliptin is vildagliptin.

[0608] In yet another embodiment, the gliptin is saxagliptin.

[0609] In a further embodiment, the gliptin is linagliptin.

[0610] In another embodiment, the gliptin is gemigliptin.

[0611] In yet another embodiment, the gliptin is anagliptin.

[0612] In a further embodiment, the gliptin is alogliptin.

[0613] In one embodiment, the gliptin is retagliptin.

[0614] In another embodiment, the gliptin is evogliptin.

[0615] In yet another embodiment, the gliptin is trelagliptin.

[0616] In a further embodiment, the gliptin is teneligliptin.

[0617] In another embodiment, the gliptin is gosogliptin.

[0618] In yet another embodiment, the gliptin is omarigliptin.

[0619] In one embodiment, the gliptin is an inhibitor of MALT1.

[0620] In one embodiment, the gliptins are selected from one or more of: vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin or cetagliptin.

[0621] In another embodiment, the gliptins are selected from one or more of: vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin.

[0622] In another embodiment, the gliptin are selected from one or more of: vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin.

[0623] In another embodiment, the gliptin are selected from one or more of: linagliptin, vildagliptin, saxagliptin or alogliptin.

[0624] In yet another embodiment, the gliptin is not sitagliptin.

[0625] Structures of gliptins are shown below:

[0626]

[0627]

[0628]

[0629]

[0630]

[0631] In another embodiment, the gliptin is preferably an inhibitor of MALT1. In a preferred embodiment, the gliptin is an allosteric inhibitor of MALT1.

[0632] In one embodiment, the multiple myeloma disease is mediated by or dependent on MALT1 activity.

[0633] In one embodiment, the method comprises an oral administration of the pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0634] In one embodiment, the method comprises an oral administration of an effective dose of the pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0635] In another embodiment, the effective dose of gliptin, preferably the once daily dose of the gliptin is preferably less than 1000 mg, preferable less than 500 mg, less than 250 mg, less than 200 mg, less than 150 mg, or equal to or less than 100 mg, even more preferably, equal to or less than 50 mg, 25mg, 20 mg, 12,5 mg, 12 mg, 10 mg, 6,25 mg, 5 mg or 2,5 mg. For some gliptins, preferred (once daily) doses are listed in Table A here below.

[0636] TABLE A: Examples of doses for a series of gliptins for use according to the invention

[0637]

[0638] In one embodiment, the method results in one or more of: inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside myeloma cells; or inhibition of MALT1 -mediated activation of NF-KB signaling pathway; or inhibition of cytokines regulated by MALT1 -mediated NF-KB signaling, such as TNF-a and / or IL-6.

[0639] In another embodiment, the method results in inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside myeloma cells.

[0640] In yet another embodiment, the method results in inhibition of MALT1 -mediated activation of NF-KB signaling pathway.

[0641] In yet another embodiment, the method results in inhibition of cytokines regulated by MALT1-mediated NF-KB signaling, wherein the cytokines such as TNF-a and / or IL-6. In a preferred embodiment, the method results in inhibition of TNF-a and / or IL-6 cytokines, which are regulated by MALT1 -mediated NF-KB signaling.

[0642] In one embodiment, the method comprises the step of diagnosing a subject as suffering from or being at risk of suffering from multiple myeloma disease, mediated by or dependent on MALT1 activity.

[0643] The aspects and / or embodiments according to the present disclosure is to be understood as illustrative of the present invention and are not intended to limit the scope of the protection. It is also to be understood that the aspects and / or embodiments defined herein may be used independently or in conjunction with any of definition, claims, drawings, other features, aspects and / or embodiments described herein. The invention contemplates all possible combinations and permutations of the various independently described aspects and / or embodiments.In Summary:

[0644] An aspect of the disclosure relates to a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating a MALT-1 mediated disease selected from multiple myeloma, in a subject, preferably a human subject, in need thereof.

[0645] An embodiment is the pharmaceutical composition for use, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin or cetagliptin, preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, more preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin or alogliptin, most preferably wherein the gliptin is sitagliptin.

[0646] An embodiment is the pharmaceutical composition for use, wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, or cetagliptin, preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin or alogliptin.

[0647] An embodiment is the pharmaceutical composition for use, wherein the gliptin is not sitagliptin. An embodiment is the pharmaceutical composition for use, wherein the gliptin is an inhibitor of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1), preferably an allosteric inhibitor of MALT1.

[0648] An embodiment is the pharmaceutical composition for use, wherein the therapeutic effect of the gliptin comprises and / or results from inhibition of MALT1 , preferably wherein the inhibition of MALT1 is an allosteric inhibition resulting from binding of the gliptin to an allosteric site within MALT1.

[0649] An embodiment is the pharmaceutical composition for use, wherein the method comprises oral administration of the pharmaceutical composition.

[0650] An embodiment is the pharmaceutical composition for use, wherein the method results in inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside myeloma cells, or inhibition of MALT1 -mediated activation of NF-KB signaling pathway, or inhibition of cytokines regulated by MALT1 -mediated NF-KB signaling, such as TNF-a and / or IL-6.

[0651] An embodiment is the pharmaceutical composition for use, wherein the MALT-1 mediated disease selected from multiple myeloma is mediated by or dependent on MALT1 activity.

[0652] An embodiment is the pharmaceutical composition for use, wherein the method comprises the step of diagnosing a subject as suffering from the MALT-1 mediated disease selected from multiple myeloma, or being at risk of suffering from the MALT-1 mediated disease selected from multiple myeloma, mediated by or dependent on MALT1 activity.An embodiment is the pharmaceutical composition for use, wherein the treatment is followed by radiotherapy and / or chemotherapy.

[0653] An embodiment is the pharmaceutical composition for use, wherein MALT1 expression status, and optionally DPP-4 expression status, comprises RNA expression analysis and / or protein expression analysis, preferably by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and / or immunoassay.

[0654] An embodiment is the pharmaceutical composition for use, wherein the method comprises the oral administration of the gliptin, pharmaceutically acceptable salt thereof or hydrate thereof, at a dose of 0.01 mg to 1000 mg per day.

[0655] An embodiment is the pharmaceutical composition for use, wherein the pharmaceutical composition is provided in an oral unit dosage form, preferably in the form of a tablet, comprising the gliptin, the pharmaceutically acceptable salt thereof or hydrate thereof, in an amount of 0.01 mg to 1000 mg.

[0656] An aspect of the disclosure relates to a therapeutic or prophylactic method of treating a MALT-1 mediated disease selected from multiple myeloma, in a subject, preferably human subject, in need thereof, said method comprising the step of administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0657] An aspect of the disclosure relates to a use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of a MALT-1 mediated disease selected from multiple myeloma, in a subject, preferably human subject, in need thereof.

[0658] EXEMPLIFYING NUMBERED EMBODIMENTS 1-15 relating to a therapeutic or prophylactic method of treating allergic inflammatory disease

[0659] Embodiment 1 : A pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating allergic inflammatory disease in a subject, preferably a human subject, in need thereof.

[0660] Embodiment 2: A pharmaceutical composition for use according to embodiment 1 , wherein the gliptin is Sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin, Alogliptin, Teneligliptin, Trelagliptin, Omarigliptin, Evogliptin, Gosogliptin, Retagliptin, Denagliptin, Cofrogliptin, Fotagliptin, Prusogliptin, Dutogliptin, Neogliptin or Cetagliptin, preferably wherein the gliptin is Sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin, Alogliptin, Teneligliptin, Trelagliptin, Omarigliptin, Evogliptin, Gosogliptin, Retagliptin, Denagliptin, Cofrogliptin, Fotagliptin, Prusogliptin, more preferably wherein the gliptin is sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin,Anagliptin or Alogliptin, more preferably wherein the gliptin is Sitagliptin, Linagliptin, Vildagliptin, Saxagliptin or Alogliptin, most preferably wherein the gliptin is sitagliptin.

[0661] Embodiment 3: A pharmaceutical composition for use according to embodiment 1 , wherein the gliptin is Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin, Alogliptin, Teneligliptin, Trelagliptin, Omarigliptin, Evogliptin, Gosogliptin, Retagliptin, Dutogliptin, Neogliptin, Denagliptin, Cofrogliptin, Fotagliptin, Prusogliptin, or Cetagliptin, preferably wherein the gliptin is Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin or Alogliptin, more preferably wherein the gliptin is Vildagliptin, Saxagliptin, Linagliptin or Alogliptin.

[0662] Embodiment 4: A pharmaceutical composition for use according to embodiment 1 , wherein the gliptin is not Sitagliptin.

[0663] Embodiment 5: A pharmaceutical composition for use according to embodiment 1 , wherein the gliptin is an inhibitor of MALT1 , preferably an allosteric inhibitor of MALT1 .

[0664] Embodiment 6: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 1-5, wherein the method comprises oral administration of the pharmaceutical composition.

[0665] Embodiment 7: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 1-6, wherein the method results in inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside aberrant cells, preferably aberrant Th2 cells.

[0666] Embodiment 8: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 1-7, wherein the allergic inflammatory disease is a chronic allergic condition and / or a late-phase allergic condition.

[0667] Embodiment 9: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 1-8, wherein the allergic inflammatory disease is selected from the group consisting of allergic asthma, anaphylaxis, sinusitis, allergic rhinitis, eczema, hives, food allergy, chronic urticaria, atopic dermatitis and ocular allergic diseases.

[0668] Embodiment 10: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 1-9, wherein the allergic inflammatory disease is mediated by or dependent on MALT1 activity.

[0669] Embodiment 11 : A pharmaceutical composition for use according to any one of the preceding numbered embodiments 1-10, wherein the method comprises the step of diagnosing a subject as suffering from or being at risk of suffering from allergic inflammatory disease mediated by or dependent on MALT1 activity.Embodiment 12: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 1-11 , wherein the method comprises the oral administration of the gliptin, pharmaceutically acceptable salt thereof or hydrate thereof, at a dose of 0.01 mg to 1000 mg per day.

[0670] Embodiment 13: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 1-12, wherein the pharmaceutical composition is provided in an oral unit dosage form, preferably in the form of a tablet, comprising the gliptin, the pharmaceutically acceptable salt thereof or hydrate thereof, in an amount of 0.01 mg to 1000 mg.

[0671] Embodiment 14: A therapeutic or prophylactic method of treating allergic inflammatory disease in a subject, preferably human subject, in need thereof, said method comprising the administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0672] Embodiment 15: Use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of allergic inflammatory disease in a subject, preferably human subject, in need thereof.

[0673] EXEMPLIFYING NUMBERED EMBODIMENTS 16-30 relating to a therapeutic or prophylactic method of treating an autoimmune disease, preferably MS

[0674] EMBODIMENT 16: A pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating multiple sclerosis (MS) in a human subject in need thereof.

[0675] Embodiment 17: A pharmaceutical composition for use according to embodiment 16, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin or cetagliptin, preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, more preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin or alogliptin, most preferably wherein the gliptin is sitagliptin.Embodiment 18: A pharmaceutical composition for use according to embodiment 16, wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, or cetagliptin, preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin or alogliptin.

[0676] Embodiment 19: A pharmaceutical composition for use according to embodiment 16, wherein the gliptin is not sitagliptin.

[0677] Embodiment 20: A pharmaceutical composition for use according to embodiment 16, wherein the gliptin is an inhibitor of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1), preferably an allosteric inhibitor of MALT1 .

[0678] Embodiment 21 : A pharmaceutical composition for use according to any one of the preceding numbered embodiments 16-20 relating to the (prophylactic) treatment of an autoimmune disease, preferably MS, wherein the method comprises oral administration of the pharmaceutical composition.

[0679] Embodiment 22: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 16-21 , wherein the method results in inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside aberrant cells, preferably aberrant ? helper 1 cells and / or T helper 17 (Th17) cells.

[0680] Embodiment 23: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 16-22, wherein the MS is clinically isolated syndrome (CIS), relapsing-remitting MS (RRMS), secondary progressive MS (SPMS), primary progressive MS (PPMS) or progressive relapsing MS (PRMS).

[0681] Embodiment 24: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 16-23, wherein the MS is at pre-symptomatic stage, clinically isolated syndrome stage, relapsing-remitting stage, or secondary progressive stage.

[0682] Embodiment 25: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 16-24, wherein the MS is mediated by or dependent on MALT1 activity.

[0683] Embodiment 26: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 16-25, wherein the method comprises the step of diagnosing a subject as suffering from or being at risk of suffering from MS mediated by or dependent on MALT1 activity.Embodiment 27: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 16-26, wherein the method comprises the oral administration of the gliptin, pharmaceutically acceptable salt thereof or hydrate thereof, at a dose of 0.01 mg to 1000 mg per day.

[0684] Embodiment 28: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 16-27, wherein the pharmaceutical composition is provided in an oral unit dosage form, preferably in the form of a tablet, comprising the gliptin, the pharmaceutically acceptable salt thereof or hydrate thereof, in an amount of 0.01 mg to 1000 mg.

[0685] Embodiment 29: A therapeutic or prophylactic method of treating MS in a human subject in need thereof, said method comprising the administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0686] Embodiment 30: Use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of MS in a human subject in need thereof.

[0687] EXEMPLIFYING NUMBERED EMBODIMENTS 31-41 relating to a therapeutic or prophylactic method of treating a cancer, preferably multiple myeloma

[0688] Embodiment 31 : A pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic or prophylactic method of treating multiple myeloma disease in a human subject in need thereof.

[0689] Embodiment 32: The pharmaceutical composition for use according to embodiment 31 , wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin or cetagliptin, preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, more preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin or alogliptin, most preferably wherein the gliptin is sitagliptin.

[0690] Embodiment 33: The pharmaceutical composition for use according to embodiment 31 , wherein the gliptin is an inhibitor of MALT1 , preferably an allosteric inhibitor of MALT1.Embodiment 34: The pharmaceutical composition for use according to any one of the preceding numbered embodiments 31-33, wherein the method comprises oral administration of the pharmaceutical composition.

[0691] Embodiment 35: The pharmaceutical composition for use according to any one of the preceding numbered embodiments 31-34, wherein the method results in one or more of: inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside myeloma cells; or inhibition of MALT1-mediated activation of NF-KB signaling pathway; or inhibition of cytokines regulated by MALT1 -mediated NF-KB signaling, such as TNF-a and / or IL-6.

[0692] Embodiment 36: The pharmaceutical composition for use according to any one of the preceding numbered embodiments 31-35, wherein the multiple myeloma disease is mediated by or dependent on MALT1 activity.

[0693] Embodiment 37: The pharmaceutical composition for use according to any one of the preceding numbered embodiments 31-36, wherein the method comprises the step of diagnosing a subject as suffering from or being at risk of suffering from multiple myeloma disease, mediated by or dependent on MALT1 activity.

[0694] Embodiment 38: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 31-37, wherein the method comprises the oral administration of the gliptin, pharmaceutically acceptable salt thereof or hydrate thereof, at a dose of 0.01 mg to 1000 mg per day.

[0695] Embodiment 39: A pharmaceutical composition for use according to any one of the preceding numbered embodiments 31-38, wherein the pharmaceutical composition is provided in an oral unit dosage form, preferably in the form of a tablet, comprising the gliptin, the pharmaceutically acceptable salt thereof or hydrate thereof, in an amount of 0.01 mg to 1000 mg.

[0696] Embodiment 40: A therapeutic or prophylactic method of treating multiple myeloma disease in a human subject in need thereof, said method comprising the (step of) administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

[0697] Embodiment 41 : Use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of multiple myeloma disease in a human subject in need thereof.EXEMPLIFYING NUMBERED EMBODIMENTS 42-61 relating to a therapeutic or prophylactic method of treating a neurological cancer, preferably glioma

[0698] EMBODIMENT 42: A pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, for use in a therapeutic method of treating neurological cancer in a human subject in need of the treatment, wherein the neurological cancer is positive for expression of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1).

[0699] Embodiment 43: Pharmaceutical composition for use according to embodiment 42, wherein the therapeutic effect of the gliptin comprises and / or results from inhibition of MALT1 , preferably wherein the inhibition of MALT1 is an allosteric inhibition resulting from binding of the gliptin to an allosteric site within MALT1.

[0700] Embodiment 44: Pharmaceutical composition for use according to any one of the preceding numbered embodiments 42-43, wherein the cancer is negative for expression of dipeptidyl peptidase-4 (DPP-4) or comprises DPP-4 expression considered as unchanged to or lower as compared to a healthy reference tissue, preferably being the tissue from which the cancer originated and / or non-cancerous tissue surrounding the cells of the cancer, advantageously being a healthy tissue of the subject.

[0701] Embodiment 45: Pharmaceutical composition for use according to any one of the preceding numbered embodiments 42-44, wherein MALT1 expression in the cancer is assessed as MALT1 overexpression.

[0702] Embodiment 46: Pharmaceutical composition for use according to any one of the preceding numbered embodiments 42-45, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin;

[0703] preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin;

[0704] more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin, and alogliptin, most preferably wherein the gliptin is sitagliptin.

[0705] Embodiment 47: Pharmaceutical composition for use according to any one of the preceding numbered embodiments 42-46, wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, and cetagliptin;

[0706] preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin;more preferably wherein the gliptin is of vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin;

[0707] even more preferably of linagliptin, vildagliptin, saxagliptin and alogliptin,

[0708] most preferably wherein the gliptin is linagliptin.

[0709] Embodiment 48: Pharmaceutical composition for use according to any one of the preceding numbered embodiments 42-47, wherein the neurological cancer is brain cancer and / or is glioma, neuroblastoma, or medulloblastoma; preferably being glioma or neuroblastoma, more preferably being glioblastoma or gliosarcoma, even more preferably being malignant glioblastoma, most preferably being glioblastoma multiforme.

[0710] Embodiment 49: Pharmaceutical composition for use according to any one of the preceding numbered embodiments 42-48, wherein the treatment comprises administering to the human subject of an effective amount of the compound, preferably directly into the cancer or into the cavity remaining after surgical removal of the cancer, possibly in combination with one or more chemotherapeutic agent and / or anticancer agent specific to the cancer of the subject.

[0711] Embodiment 50: Pharmaceutical composition for use according to any one of the preceding numbered embodiments 42-49, wherein the treatment is followed by radiotherapy and / or chemotherapy.

[0712] Embodiment 51 : Pharmaceutical composition for use according to any one of the preceding numbered embodiments 42-50, wherein MALT1 expression status, and optionally DPP-4 expression status, comprises RNA expression analysis and / or protein expression analysis, preferably by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and / or immunoassay.

[0713] Embodiment 52: An in vitro method for determining an increased probability that a neurological cancer in a human subject will respond to the treatment with

[0714] a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, wherein the gliptin is selected from wherein the gliptin is selected from any one or more of sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, , cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin, and cetagliptin; the method comprising the steps of:

[0715] - providing a test first sample obtained from the subject;

[0716] - detecting the presence of MALT1 expression and preferably assessing the expression level of MALT 1 in the first test sample;

[0717] - preferably, comparing the assessed expression level of MALT1 to a reference MALT1 expression level;

[0718] wherein finding that MALT1 expression is present in the first test sample and preferably that the expression level of MALT1 assessed in the first test sample is higher as compared to the referenceMALT1 expression level is indicative of an increased probability that the subject will respond to the treatment.

[0719] Embodiment 53: The in vitro method according to embodiment 52, further comprising the steps of: - providing a second test sample obtained from the subject;

[0720] - detecting the presence of DPP-4 expression or optionally assessing the expression level of DPP-4 or DPP-4 enzymatic activity level in the second test sample;

[0721] - preferably, comparing the assessed expression level or enzymatic activity level of DPP-4 to a reference DPP-4 expression level or a reference DPP-4 enzymatic activity level, respectively; wherein finding that DPP-4 expression is absent in the second test sample or optionally that the expression level of DPP-4 or DPP-4 enzymatic activity level assessed in the second test sample is lower as compared to the reference DPP-4 expression level or a reference DPP-4 enzymatic activity level, respectively; preferably finding that DPP-4 expression is absent in the second test sample is indicative of an increased probability that the subject will respond to the treatment, wherein the response is further associated with an increased probability of not causing adverse DPP-4-inhibition-associated side effects.

[0722] Embodiment 54: The in vitro method according to any one of the numbered embodiments 52-53, wherein the first test sample and / or the second test sample is or comprises biopsy sample, cerebrospinal fluid (CSF) sample, or a blood sample, preferably being biopsy sample or the cerebrospinal fluid (CSF) sample, even more preferably being biopsy sample.

[0723] Embodiment 55: The in vitro method according to any one of the numbered embodiments 52-54, wherein detecting the presence of expression and / or assessing the expression level comprises RNA analysis, preferably comprising RNA-sequencing or reverse transcription-quantitative polymerase chain reaction (RT-qPCR), more preferably comprising RT-qPCR.

[0724] Embodiment 56: The in vitro method according to any one of the numbered embodiments 52-55, wherein detecting the presence of expression and / or assessing the expression level comprises protein analysis, preferably comprising an immunoassay, more preferably comprising ELISA.

[0725] Embodiment 57: The in vitro method according to any one of the numbered embodiments 52-56, wherein the first test sample and the second test sample are the same test sample, preferably wherein detecting the presence of MALT1 and DPP-4 expression, optionally assessing the expression level of MALT1 and DPP-4 is performed as part of the same assay, more preferably being a multiplex RT-qPCR assay or an immunoassay.

[0726] Embodiment 58: The in vitro method according to any one of the numbered embodiments 52-57, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin,teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin;

[0727] preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin;

[0728] more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin, and alogliptin, most preferably wherein the gliptin is sitagliptin;

[0729] and / or

[0730] wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, and cetagliptin;

[0731] preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin;

[0732] more preferably wherein the gliptin is of vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin;

[0733] even more preferably of linagliptin, vildagliptin, saxagliptin and alogliptin,

[0734] most preferably wherein the gliptin is linagliptin.

[0735] Embodiment 59: The in vitro method according to any one of the numbered embodiments 52-58, wherein the reference MALT1 expression value, and optionally the reference DPP-4 expression value, is defined and has (have) been predetermined based on assessing MALT1 expression levels, and optionally DPP-4 expression levels, respectively, in a plurality of control samples obtained from healthy control subjects.

[0736] Embodiment 60: The in vitro method according to any one of the numbered embodiments 52-58, wherein the reference MALT1 expression value, and optionally the reference DPP-4 expression value, is determined based on assessing normal MALT1 expression levels, and optionally normal DPP-4 expression levels, respectively, in a healthy tissue sample obtained from the subject, preferably being a healthy tissue corresponding to the tissue from which the cancer originated, optionally being healthy non-cancerous tissue surrounding the cancer cells.

[0737] Embodiment 61 : The in vitro method according to any one of the one of the numbered embodiments 52-59, wherein the expression level of MALT1 assessed for the first test sample is at least 2 fold higher than the reference MALT1 expression value, preferably being at least 5 fold higher; more preferably being at least 10 fold higher.

[0738] EXAMPLESThe following examples serve to illustrate the (intracellular) allosteric MALT1 inhibition provided by gliptins, i.e. DPP-4 inhibitors, according to the present invention, preferably sitagliptin.

[0739] These examples 1-4 show that DPP-4 inhibitors bind allosterically to MALT1 and inhibit the enzymatic activity of MALT1 which in turn reduces Th2 cell activation by acting on T-cell receptor (TCR)-mediated NF-KB pathway.

[0740] Example 1: Sitagliptin inhibits the proteolytic activity of MALT1

[0741] Inhibition of MALT1 proteolytic activity by sitagliptin is assessed. As positive controls, two MALT1 inhibitors were included, namely MI-2 and (S)-mepazine. Each of the compound is used at a concentration of 10 pM.

[0742] MI-2 (C19H17CI3N4O3; 2-chloro-N-[4-[5-(3,4-dichlorophenyl)-3-(2-methoxyethoxy)-1 ,2,4-triazol-1-yl] phenyl] acetamide) is an orthosteric and irreversible MALT1 inhibitor (Fontan et al, 2012).

[0743] (S)-mepazine (C19H22N2S; 10-{[(3S)-1 -Methyl-3-piperidinyl] methyl}-10H-phenothiazine) is an allosteric MALT1 inhibitor with higher binding affinity and inhibitory activity than (R)-mepazine or (±)-mepazine (Schlauderer et al, 2013).

[0744] A so-called buffer only test-condition (i.e. MALT1 + Ac-LRSR-AMC substrate + buffer) served as negative control.

[0745] The MALT1 activity is measured according to the activity assay developed by Fontan et al., 2012 where the MALT1 substrate peptide LRSR is linked to the fluorogen AMC (7-amino-4-methylcoumarin). Cleavage of the Ac-LRSR-AMC substrate by MALT1 results in release of AMC and a fluorescent signal. The recombinant full-length human MALT1 and Ac-LRSR-AMC - with excitation / emission wavelengths of 360 / 460 nm in black, low binding microtiter plates with a microplate reader are used for monitoring fluorogenic reactions. To eliminate false positives due to compound autofluorescence, two time points were measured for each reaction and hence, the fluorescence difference between such time points (T2-T1) was defined as MALT1 activity.

[0746] The final percent inhibition was calculated with the formula:

[0747] [fluorescence_test compound (T2-T1) - fluorescence_negative control (T2-T1)]

[0748] _ x 100 [fluorescence_positive control (T2-T1) - fluorescence_negative control (T2-T1)]

[0749] Sitagliptin was identified as a MALT1 inhibitor using 40% inhibition as a threshold and validated against MI-2 and / or (S)-mepazine.

[0750] EC50 (effective concentration 50%) refers to the concentration of a test-compound at which 50% of its maximum effect is achieved. The term is quite general, regardless of whether a test-compound enhances or diminishes the biological parameter in question. Its equation is a modified version of the IC50 equation, taking the ratio of the response (R) to the maximum response (Rmax): {R / Rmax} = {1 / [1 +(E50 / l)n]}. In cases where a test-compound completely inhibits a biological activity at high dose, thevalues of EC50 and IC50 are identical (where Rmax = 100%). However, some test-compounds achieve only partial attenuation of a biological activity, even at high concentrations. In this case, IC50 values may be misleading about potency. Furthermore, efficacious test-compounds have more similar IC50 and EC50 values than their less efficacious counterparts. Of note and in contrast to IC50 values, EC50 values report on the binding affinity of the test-compound (effective concentration 50% is achieved when half of the target protein is bound). Using EC50 values allows the dose-response of test-compounds to be best quantified and reported when complex biological systems are employed, such as cell or animal models. Cell lines were in log phase of growth, harvested and seeded in a 96-well plate at optimum plating density. Next, cells were incubated with and without test-compounds (0.1 pM - 1 pM - 10 pM) in the presence of positive, negative and untreated controls. EC50 values were determined by Trypan blue dye-exclusion at t=48h and the IC50 calculator (Quest Graph™ IC50 Calculator." AAT Bioquest, Inc., 25 Apr. 2023, https: / / www.aatbio.com / tools / ic50-calculator).

[0751] Sitagliptin inhibits the proteolytic activity of MALT1 with an EC50 value of 1.3 pM (Table 1). Results are mean ± standard deviation of three independent experiments done in triplicates.

[0752] Table 1. Sitagliptin, MI-2 and (S)-mepazine EC50 values

[0753]

[0754] Example 2: Enzyme (catalytic) activity of the CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 isoenzymes

[0755] CYP450s are the major mediators of xenobiotics’ toxicity and interactions. Ideally, someone would like to have xenobiotics that do not induce or inhibit those enzymes. In a nutshell, here presented data show that these enzymes function properly and unaltered (metabolic and catalytic activities) under influence of exposure to gliptins and when inhibition is considered, sitagliptin has the most favourable profile among gliptins but also when compared to mepazine. CYP2D6 and CYP3A4 inhibition is the most detrimental effect of mepazine as those enzymes are involved in most xenobiotics’ metabolism to date. The enzymes selected for testing of influence of gliptins and mepazine are the seven most relevant enzymes of the kind in humans.

[0756] To determine the isozyme-specific CYP450-metabolism, baculosomes® expressing CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 human CYP450 isoenzymes were purchased from Thermo Fisher Scientific (Waltham, MA, USA). All reagents were handled and prepared according to the manufacturer’s protocol. Sitagliptin was tested at 1 pM. Enzymatic activity in the presence of sitagliptin was assessed based on the kinetic model for CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 isoforms. Reactions took place in triplicates at 20°C.Glucose-6-phosphate at 333 mM and glucose-6-phosphate-dehydrogenase at 30 U / mL in 100 mM potassium phosphate, pH 8.0 (regeneration system) converted NADP+ (10 mM in 100 mM potassium phosphate, pH 8.0) into NADPH to initiate the CYP450 reaction. Upon the addition of the fluorescent substrate, immediately (in the presence of test-compound; A is the rate observed in the presence of negative (solvent, DMSO) control.

[0757] The administration of test-compounds at 1 pM (60 minutes) and Relative Fluorescence Units (RFU) measured for the reaction under different conditions.

[0758] CYP450 (%) inhibition of CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 isoenzymes were measured upon the administration of test compounds at 1 pM (60 minutes).

[0759] Data show that sitagliptin, saxagliptin, alogliptin and linagliptin do not inhibit nor induces CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 human CYP450 isoenzymes.

[0760] It was apparent that mepazine showed substantial influence on CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 isoenzymes, specifically on CYP2D6.

[0761] Example 3: qliptins are allosteric inhibitors of MALT1

[0762] MALT1 has been co-crystallized with MLT-748, a 5 nM inhibitor, which binds to an allosteric pocket through various H-bond and hydrophobic interactions (Quancard et al., 2019). The crystal structure of human MALT1 (PDB: 6H4A) (Quancard et al., 2019) was obtained from protein data bank and used for docking calculations of the co-crystallized ligand MLT-748 as a reference. In addition, the docking calculations were performed with the same MALT1 structure and mepazine, a known allosteric inhibitor of MALT1 , and sitagliptin, retagliptin, evogliptin, gemigliptin, fotagliptin, linagliptin, trelagliptin, denagliptin, tenegliptin, gosogliptin, alogliptin, anagliptin, prusogliptin, cofrogliptin, omarigliptin, saxagliptin, vildagliptin. The software used for the dockings was gnina (McNutt et al.).

[0763] As a negative control, the docking of the non-MALT1 inhibiting compound VI (Tran et al., 2019; molecule 11) was included in the calculations. Indeed, referring to Table A1 , an expected low docking score was apparent.

[0764] Results show that for MLT-748, as in the crystal structure the triazole nitrogen of MLT-748 interacts through a hydrogen bond with N393 and the two scaffold nitrogens of MLT-748 form hydrogen bonds with the E397 sidechain oxygens as in the crystal structure (Figure 1A). In addition, MLT-748 forms several hydrophobic interactions with aliphatic residues L383, L346, V381 and L401 of MALT1. Surprisingly, the docking mode of sitagliptin to MALT-1 demonstrates a close to identical profile of interactions as seen for MLT-748 binding to MALT1 (Figure 1 B). The protonated amide group of sitagliptin forms a salt bridge with E397 and potentially interacts with the backbone oxygen of A394. The trifluoro aromatic ring is positioned similarly to the MLT-748 chloropyridine in a hydrophobic groove consisting ofV381 , L383, L386, A394 and 1712. The meta positioned fluorine is pointing into the pocket in a similar manner as the chloropyridine chlorine. The triazolopyrazine behaves similarly to MLT-748 chloropyrazolopyrimidine, as it forms hydrophobic interactions with V381 , L401 and W580. The trifluoro group with its unique hydrophobic properties interacts with W580 and the aliphatic chain of K397.Even more surprisingly, the gliptins Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Anagliptin and Alogliptin demonstrate similar docking scores, indicative for their allosteric binding to MALT1 similarly as has been demonstrated for mepazine and MLT-748.

[0765] Docking scores (higher is better) of MLT-748, mepazine, known allosteric inhibitors of MALT1 , and sitagliptin, retagliptin, evogliptin, gemigliptin, fotagliptin, linagliptin, trelagliptin, denagliptin, tenegliptin, gosogliptin, alogliptin, anagliptin, prusogliptin, cofrogliptin, omarigliptin, saxagliptin, vildagliptin, are highlighted in table A1 here below, showing that based on these results all indicated molecules including the series of gliptins, are allosteric inhibitors of MALT1.

[0766] Table A1. Docking scores of MLT-748, mepazine, known allosteric inhibitors of MALT1 , and now newly identified allosteric inhibitors of MALT1 sitagliptin, retagliptin, evogliptin, gemigliptin, fotagliptin, linagliptin, trelagliptin, denagliptin, tenegliptin, gosogliptin, alogliptin, anagliptin, prusogliptin, cofrogliptin, omarigliptin, saxagliptin, vildagliptin in the allosteric site of MALT1

[0767]

[0768] Example 4: Sitagliptin decreases cytokine production of Th2-cellsCell line model for allergic inflammation

[0769] Fortesting allosteric inhibitors of MALT1 in allergic inflammation, choosing cell lines with active MALT1 expression and involvement in allergic and inflammatory pathways is key.

[0770] Jurkat T Cells: This T-cell line is widely used to study MALT1 due to its active expression in T-cell receptor signaling. Testing MALT1 allosteric inhibitors in Jurkat cells can help analyze effects on T-cell-mediated aspects of allergic inflammation.

[0771] Jurkat T cells can be used as a model to study aspects of allergic inflammation, particularly the role of T-cell signaling and cytokine production. While allergic inflammation primarily involves mast cells, basophils, and eosinophils, T cells play a significant regulatory role in the immune response, especially through Th2-mediated cytokine production (e.g., IL-4, IL-5, and IL-13).

[0772] Modeling T-Cell Receptor (TCR) Signaling:

[0773] ■ Jurkat cells are a well-established model for studying TCR-mediated activation, which is crucial in the immune responses driving allergic inflammation.

[0774] ■ Allergic conditions are often characterized by excessive T-cell activation and dysregulated cytokine production, processes that can be modeled in Jurkat cells.

[0775] Cytokine Analysis:

[0776] ■ Jurkat cells produce cytokines like IL-2 and IL-4 upon stimulation, making them a useful model for studying inflammatory signaling pathways.

[0777] ■ The Jurkat cells do not fully mimic Th2 cells (key in allergies), nonetheless they provide insights into general T-cell activation and cytokine production.

[0778] Testing Therapeutic Interventions:

[0779] Jurkat cells are commonly used for screening the effects of small molecules, including inhibitors of MALT 1 , NF-KB, or other pathways involved in allergic inflammation. Accessible and Reproducible: Jurkat cells are easy to culture and highly responsive to TCR stimulation, making them a convenient model for in vitro studies of immune responses.

[0780] Expression characteristics

[0781] The Jurkat T cells express MALT1 , making them one of the primary cell lines used to study MALT1 function and inhibition. MALT1 is a crucial component in T-cell receptor signalling, where it forms part of the CBM complex (composed of CARMAI , BCL10, and MALT1), which is essential for activating NF-KB and other downstream immune response pathways. This makes Jurkat cells a valuable model for examining MALT1 -mediated signaling and the effects of MALT1 inhibitors in T-cell-driven conditions. The Jurkat T cells do not show DPP4 expression or only have a low expression / undetectable level of DPP4, thereby excluding effects which may be related to a DPP4 targeting of the tested gliptins.

[0782] Cell CultureJurkat T cells (ATCC TIB-152) were used to study the effects of MALT1 allosteric inhibitors in allergic inflammation. The cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, and 1% penicillin-streptomycin. The cells were maintained at 37°C in a humidified atmosphere with 5% CO2and were routinely kept at a density between 0.2 x 106and 1 x 106cells / mL. Cells were passaged every 2-3 days to maintain exponential growth. Cell cultures were performed in 3D.

[0783] Treatment with MALT1 Allosteric Inhibitor

[0784] Sitagliptin was prepared in DMSO and stored at -20°C. Working dilutions were freshly prepared in culture medium with final concentrations ranging from 0.1 pM to 100 pM. Control groups included a vehicle control (0.1% DMSO) and a positive control using a known MALT1 inhibitor, namely (S)-mepazine. (S)-mepazine (C19H22N2S; 10-{[(3S)-1 -Methyl-3-piperidinyl] methyl}-10H-phenothiazine) is an allosteric MALT1 inhibitor with higher binding affinity and inhibitory activity than (R)-mepazine or (±)-mepazine (Schlauderer et al, 2013). Jurkat T cells were in log phase of growth, harvested and seeded in a 96-well plate at optimum plating density. Next, sitagliptin and positive control, (S)-mepazine were tested at 0.1 pM, 1 pM, 10 pM and 100 pM at t=48h. DMSO was <0.05%.

[0785] Stimulation of Inflammatory Pathways

[0786] Jurkat T cells were stimulated with 20 ng / mL phorbol 12-myristate 13-acetate (PMA) and 1 pM ionomycin to mimic T-cell activation. Additionally, recombinant human IL-4 (20 ng / mL) was added to induce Th2 cytokine pathways.

[0787] Assessment of MALT1 Inhibition

[0788] To evaluate the effects of sitagliptin (MALT1 allosteric inhibitor), first, NF-KB activity was assessed along with cytokine secretion. Cytokines (IL-4, IL-13) were quantified in cell culture supernatants collected 24 hours after stimulation using commercially available ELISA kits according to the manufacturer’s instructions and normalized to cell viability.

[0789] Functional Assays

[0790] Cell proliferation and metabolism were assessed using the Alamar blue assay, a non-destructive and sensitive method to evaluate metabolic activity in viable cells. Jurkat T cells were seeded in 96-well plates at a density of 5 x 104cells per well in 100 pL of culture medium and treated with sitagliptin at various concentrations for 24-48 hours. Following the incubation, 10 pL of Alamar Blue reagent (10% of total volume) was added to each well, and the plates were incubated at 37°C for 4-6 hours. Fluorescence was measured using a microplate reader with an excitation / emission wavelength of 570 / 600 nm. Untreated cells were used as a positive control, and wells containing only culture medium served as a background control. The reduction in fluorescence intensity in treated wells, compared to untreated controls, was indicative of reduced cell proliferation. (S)-mepazine served as a positive control. Results were expressed as a percentage of cell viability relative to the untreated control group.Statistical Analysis

[0791] All experiments were performed in triplicate, and data were presented as the mean ± standard deviation. Statistical significance was determined using one-way ANOVA followed by Tukey’s post-hoc test, with a p-value < 0.05 considered statistically significant.

[0792] Sitagliptin-mediated inhibition of MALT1 will result in suppression of NF-KB-driven secretion of Th2 cytokines such as IL-4, and IL-13, which are critical mediators of allergic inflammation.

[0793] Sitaqliptin decreases Jurkat cell survival (allergic inflammation model)

[0794] Figure 2 shows the effects of S-mepazine and sitagliptin on Jurkat cell survival (allergic inflammation model). Results are mean ± standard deviation of three independent experiments done in triplicates. Sitagliptin and positive control and (S)-mepazine, were tested at 0.001-100 pM. DMSO was <0.05%.

[0795] Sitaqliptin decreases Jurkat cell metabolism

[0796] (S)-mepazine and sitagliptin both reduced cell metabolism in a dose-dependent manner. Figure 3 shows the percentage of cell metabolism relative to untreated controls after treatment with (S)-mepazine (positive control) and sitagliptin at various concentrations (1 pM, 10 pM, and 50 pM). Cell metabolism was assessed using the Alamar Blue assay. Error bars represent mean ± standard deviation from triplicate experiments.

[0797] Sitaqliptin decreases cytokine production

[0798] Inhibition of MALT1 should reduce the secretion of pro-inflammatory cytokines like IL-4 and IL-13, which are major mediators of allergic inflammation. Figure 4 shows the effects of S-mepazine (5.0 pM) and sitagliptin (5.0 pM) on cytokine production, when Jurkat cells treated with MALT1 inhibitors were compared to untreated cells (no decrease in IL-4 and IL-13 production). Results are mean ± standard deviation of three independent experiments done in triplicates.

[0799] In addition, the following examples 5-7 in combination with example 3 here above show that DPP-4 inhibitors bind allosterically to MALT1 and inhibit the same which in turn reduce Th1 and Th17 cell activation by acting on T-cell receptor (TCR)-mediated NF-KB pathway whose activation causes autoimmune diseases including multiple sclerosis.

[0800] Example 5: Sitagliptin decreases Jurkat cell survival

[0801] MALT1 plays a key role in T-cell receptor (TCR)-mediated NF-KB activation. Its inhibition by sitagliptin is expected to reduce Th1 and Th17 cell activation, two subsets critical for multiple sclerosis (MS) pathogenesis.

[0802] Figure 5 shows the effects of S-mepazine and sitagliptin on Jurkat cell survival (multiple sclerosis model). Results are mean ± standard deviation of three independent experiments done in triplicates. Sitagliptin and positive control and (S)-mepazine, were tested at 0.001-100 pM. DMSO was <0.05%.

[0803] Example 6: Sitagliptin decreases cell metabolism(S)-mepazine and sitagliptin both reduced cell metabolism in a dose-dependent manner. Figure 6 shows the percentage of cell metabolism relative to untreated controls after treatment with (S)-mepazine (positive control) and sitagliptin at various concentrations (1 pM, 10 pM, and 50 pM). Cell metabolism was assessed using the Alamar Blue assay. Error bars represent mean ± standard deviation from triplicate experiments.

[0804] Example 7: Sitagliptin decreases cytokine production

[0805] Inhibition of MALT1 should reduce the secretion of pro-inflammatory cytokines like IL-17 and IFN-y (suppress NF-KB activity), which are major drivers of autoimmune inflammation in MS.

[0806] Figure 7 shows the effects of S-mepazine (8 pM) and sitagliptin (8 pM) on cytokine production, when Jurkat cells treated with MALT1 inhibitors were compared to untreated cells (no decrease in IL-17 and IFN-y production). Results are mean ± standard deviation of three independent experiments done in triplicates.

[0807] Materials and methods

[0808] Cell Culture

[0809] Jurkat T cells (ATCC TIB-152) were used to study the effects of MALT1 allosteric inhibitors in multiple sclerosis. The cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, and 1% penicillin-streptomycin. The cells were maintained at 37°C in a humidified atmosphere with 5% CO2and were routinely kept at a density between 0.2 x 106and 1 x 106cells / mL. Cells were passaged every 2-3 days to maintain exponential growth. Cell cultures were performed in 3D.

[0810] Treatment with MALT1 Allosteric Inhibitor

[0811] Sitagliptin was prepared in DMSO and stored at -20°C. Working dilutions were freshly prepared in culture medium with final concentrations ranging from 0.1 pM to 100 pM. Control groups included a vehicle control (0.1% DMSO) and a positive control using a known MALT1 inhibitor, namely (S)-mepazine. (S)-mepazine (C19H17CI3N4O3; 10-{[(3S)-1 -Methyl-3-piperidinyl] methyl}-10H-phenothiazine) is an allosteric MALT1 inhibitor with higher binding affinity and inhibitory activity than (R)-mepazine or (±)-mepazine (Schlauderer et al, 2013). Jurkat T cells were in log phase of growth, harvested and seeded in a 96-well plate at optimum plating density. Next, sitagliptin and positive control, (S)-mepazine were tested at 0.1 pM, 1 pM, 10 pM and 100 pM at t=48h. DMSO was <0.05%.

[0812] Stimulation of Inflammatory Pathways

[0813] For multiple sclerosis, T-cell activation was mimicked by treating cells with 1 pg / mL anti-CD3 and 1 pg / mL anti-CD28 antibodies, alongside recombinant human IL-17 (10 ng / mL) and IFN-y (20 ng / mL) to induce Th1 / Th17 cytokine responses (relevant to MS).

[0814] Assessment of MALT1 Inhibition

[0815] To evaluate the effects of sitagliptin (MALT1 allosteric inhibitor), first, NF-KB activity was assessed along with cytokine secretion. Cytokines (IL-17, and IFN-y) were quantified in cell culture supernatantscollected 24 hours after stimulation using commercially available ELISA kits according to the manufacturer’s instructions and normalized to cell viability.

[0816] Functional Assays

[0817] Cell proliferation and metabolism were assessed using the Alamar blue assay, a non-destructive and sensitive method to evaluate metabolic activity in viable cells. Jurkat T cells were seeded in 96-well plates at a density of 5 x 104cells per well in 100 pL of culture medium and treated with sitagliptin at various concentrations for 24-48 hours. Following the incubation, 10 pL of Alamar Blue reagent (10% of total volume) was added to each well, and the plates were incubated at 37°C for 4-6 hours. Fluorescence was measured using a microplate reader with an excitation / emission wavelength of 570 / 600 nm. Untreated cells were used as a positive control, and wells containing only culture medium served as a background control. The reduction in fluorescence intensity in treated wells, compared to untreated controls, was indicative of reduced cell proliferation. (S)-mepazine served as a positive control. Results were expressed as a percentage of cell viability relative to the untreated control group.

[0818] Statistical Analysis

[0819] All experiments were performed in triplicate, and data were presented as the mean ± standard deviation. Statistical significance was determined using one-way ANOVA followed by Tukey’s post-hoc test, with a p-value < 0.05 considered statistically significant.

[0820] Examples 8-12

[0821] The following examples 8-12 in combination with example 3 serve to illustrate the gliptins for use in the treatment of multiple myeloma. It further illustrates the gliptins as allosteric MALT1 inhibitors and the inhibition provided by the gliptins according to the method of treatment or prevention of present invention. In some of the examples, sitagliptin has been illustrated for testing as preferred gliptin, wherein other gliptins would also fit the same way as sitagliptin.

[0822] Materials and methods:

[0823] Cell Culture

[0824] JJN3 cells, a human multiple myeloma cell line, were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin-streptomycin, and 2 mM L-glutamine. Cells were maintained at 37°C in a humidified atmosphere with 5% CO2and passaged every 2-3 days to maintain exponential growth. Cell density was maintained between 0.2 x 106and 1 x 106cells / mL throughout the study.

[0825] Stimulation of Inflammatory Pathways

[0826] To mimic the pro-inflammatory tumor microenvironment characteristic of multiple myeloma, JJN3 cells were stimulated with recombinant human interleukin-6 (IL-6; 20 ng / mL) for 24 hours prior to treatment with MALT1 inhibitors. IL-6 stimulation is known to enhance survival signaling pathways in multiple myeloma cells, including those mediated by NF-KB.Treatment with DPP-4 inhibitor compounds through MALT1 Allosteric Inhibition

[0827] Sitagliptin was prepared in DMSO and stored at -20°C. Working dilutions were freshly prepared in culture medium with final concentrations ranging from 0.1 pM to 100 pM. Control groups included a vehicle control (0.1% DMSO) and a positive control using a known MALT1 inhibitor, namely (S)-mepazine.

[0828] S-mepazine is an allosteric MALT1 inhibitor with higher binding affinity and inhibitory activity than (R)-mepazine or (±)-mepazine (Schlauderer et al, 2013).

[0829] All experiments were performed in triplicate, and data were presented as the mean ± standard deviation. Statistical significance was determined using one-way ANOVA followed by Tukey’s post-hoc test, with a p-value < 0.05 considered statistically significant.

[0830] Example 8 - Cell survival

[0831] JJN3 cells were in log phase of growth, harvested and seeded in a 96-well plate at optimum plating density. Next, sitagliptin and positive control, S-mepazine were tested at 0.001 pM, 0.01 pM, 0.1 pM, 1 pM, 5 pM, 10 pM, 50 pM and 100 pM at t=48h. DMSO was <0.05%.

[0832] As MALT1 is involved in NF-KB activation, which supports survival and proliferation of multiple myeloma (MM) cells. Sitagliptin-mediated inhibition of MALT1 downregulated NF-KB-driven survival signals, leading to increased apoptosis and reduced proliferation of MM cells (such as JJN3 cells). Figure 8 shows the effects of S-mepazine and sitagliptin on JJN3 cell survival. Results are mean ± standard deviation of three independent experiments done in triplicates.

[0833] Example 9 - Cell viability, proliferation and apoptosis assay

[0834] Cell proliferation and metabolism were assessed using the Alamar blue assay, a non-destructive and sensitive method to evaluate metabolic activity in viable cells. JJN3 cells were seeded in 96-well plates at a density of 5 x 104cells per well in 100 pL of culture medium and treated with sitagliptin at various concentrations for 24-48 hours. Following the incubation, 10 pL of Alamar Blue reagent (10% of total volume) was added to each well, and the plates were incubated at 37°C for 4-6 hours. Fluorescence was measured using a microplate reader with an excitation / emission wavelength of 570 / 600 nm. Untreated cells were used as a positive control, and wells containing only culture medium served as a background control. The reduction in fluorescence intensity in treated wells, compared to untreated controls, was indicative of reduced cell proliferation. S-mepazine served as a positive control. Results were expressed as a percentage of cell viability relative to the untreated control group.

[0835] S-mepazine and sitagliptin both reduced cell metabolism in a dose-dependent manner. Figure 9 shows the percentage of cell metabolism relative to untreated controls after treatment with S-mepazine (positive control) and sitagliptin at various concentrations (1 pM, 10 pM, and 50 pM). Cell metabolism was assessed using the Alamar Blue assay. Error bars represent mean ± standard deviation from triplicate experiments.

[0836] Example 10 - Cytokines analysisTo evaluate the effects of sitagliptin (MALT1 allosteric inhibitor), supernatants were collected after 24 hours of treatment and analyzed for cytokines regulated by MALT1 -mediated NF-KB signaling, such as TNF-a and IL-6, using ELISA kits. Cytokine levels were measured according to the manufacturer’s instructions and normalized to cell viability.

[0837] Figure 10 shows the effects of S-mepazine (5 pM) and sitagliptin (5 pM) on cytokine production, when JJN3 cells treated with MALT1 inhibitors were compared to untreated cells (no decrease in IL-6 and TNF-a production). Sitagliptin decreases cytokine production: Inhibition of MALT1 reduced the secretion of pro-inflammatory cytokines like IL-6 and TNF-a, which are crucial for the autocrine and paracrine survival signaling in MM cells. Results are mean ± standard deviation of three independent experiments done in triplicates.

[0838] Example 11 - Proteolytic inhibition of MALT1

[0839] Sitagliptin inhibition of MALT1 proteolytic activity was performed at 10 pM. Fluorogenic reactions were monitored in the presence of recombinant full-length human MALT1 and Ac LRSR-AMC with excitation / emission wavelengths of 360 / 460 nm in black, low binding microtiter plates using a microplate reader. Cleavage of the Ac-LRSR-AMC substrate by MALT1 results in the release of AMC and a fluorescent signal. To eliminate false positives due to compound autofluorescence, two time points were measured for each reaction and hence, the fluorescence difference between such time points (T2-T1) was defined as MALT1 activity. The final percent inhibition was calculated with the formula: {[fluorescence_test compound(T2-T1) -fluorescence_negative control (T2-T1)] / [fluorescence_positive control (T2-T1) - fluorescence_negative control (T2-T1)]} x 100. (S)-mepazine or MI-2 (C19H17CI3N4O3; 2 chloro-N-[4-[5-(3,4-dichlorophenyl)-3-(2-methoxyethoxy)-1 ,2,4-triazol-1-yl] phenyl] acetamide) were used as positive controls at 10 pM. A so-called buffer only test-condition (i.e. MALT1+Ac-LRSR-AMC substrate+buffer) served as negative control. Sitagliptin was identified as a MALT1 inhibitor using 40% inhibition as a threshold and validated against MI-2 and / or S-mepazine.

[0840] EC50 (effective concentration 50%) refers to the concentration of a test-compound at which 50% of its maximum effect is achieved. The term is quite general, regardless of whether a test-compound enhances or diminishes the biological parameter in question. Its equation is a modified version of the IC50 equation, taking the ratio of the response (R) to the maximum response (Rmax): {R / Rmax} = {1 / [1 +(E50 / l)n]}. In cases where a test-compound completely inhibits a biological activity at high dose, the values of EC50 and IC50 are identical (where Rmax = 100%). However, some test-compounds achieve only partial attenuation of a biological activity, even at high concentrations. In this case, IC50 values may be misleading about potency. Furthermore, efficacious test-compounds have more similar IC50 and EC50 values than their less efficacious counterparts. Of note and in contrast to IC50 values, EC50 values report on the binding affinity of the test-compound (effective concentration 50% is achieved when half of the target protein is bound). Using EC50 values allows the dose response of test-compounds to be best quantified and reported when complex biological systems are employed, such as cell or animal models.

[0841] Cell lines were in log phase of growth, harvested and seeded in a 96-well plate at optimum plating density. Next, cells were incubated with and without test-compounds (0.1 pM - 1 pM - 10 pM) in the presence of positive, negative and untreated controls. EC50 values were determined by Trypan bluedye-exclusion at t=48h and the IC50 calculator (Quest Graph™ IC50 Calculator." AAT Bioquest, Inc., 25 Apr. 2023, https: / / www.aatbio.com / tools / ic50 calculator).

[0842] Sitagliptin inhibits the proteolytic activity of MALT1 with an EC50 value of 1 .3 pM (Table B here below). Results are mean ± standard deviation of three independent experiments done in triplicates.

[0843] TABLE B: SITAGLIPTIN, MI-2 AND (S)-MEPAZINE EC50 VALUES

[0844] Test-compounds EC50 values (pM)

[0845] Sitagliptin 1.3

[0846] MI-2 1.0

[0847] (S)-mepazine 1.0

[0848] Example 12 - Enzyme (catalytic) activity of the CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 isoenzymes

[0849] To determine the isozyme-specific CYP450-metabolism, baculosomes® expressing CYP1 A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 human CYP450 isoenzymes were purchased from Thermo Fisher Scientific (Waltham, MA, USA). All reagents were handled and prepared according to the manufacturer’s protocol. Sitagliptin was tested at 1 pM. Enzymatic activity in the presence of sitagliptin was assessed based on the kinetic model for CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 isoforms. Reactions took place in triplicates at 20°C. Glucose-6-phosphate at 333 mM and glucose-6-phosphate-dehydrogenase at 30 U / mL in 100 mM potassium phosphate, pH 8.0 (regeneration system) converted NADP+ (10 mM in 100 mM potassium phosphate, pH 8.0) into NADPH to initiate the CYP450 reaction. Upon the addition of the fluorescent substrate, immediately (in the presence of test-compound; A is the rate observed in the presence of negative (solvent, DMSO) control.

[0850] The administration of test-compounds at 1 pM (60 minutes) and Relative Fluorescence Units (RFU) measured for the reaction under different conditions.

[0851] CYP450 (%) inhibition of CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 isoenzymes were measured upon the administration of test compounds at 1 pM (60 minutes).

[0852] Data show that sitagliptin, saxagliptin, alogliptin and linagliptin do not inhibit nor induces CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 human CYP450 isoenzymes.

[0853] It was apparent that mepazine showed substantial influence on CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 isoenzymes, specifically on CYP2D6.

[0854] Overall, based on the insights gained with the examples 3 and 8-12, it is concluded that gliptins are MALT 1 inhibitors capable of killing tumor cells, as determined with computational exploration of binding capabilities of the gliptins to the allosteric binding site and as determined with confirmative in vitro cellbased assays using human cancer-patient derived multiple myeloma cells. Thus, the experimental dataalign with the computational data. Gliptins identified as allosteric MALT1 inhibitors based on computational data and based on in vitro cell-based assay data match.

[0855] EXAMPLES 13-18

[0856] The following examples 13-18, together with example 3, serve as an illustration of the described herein allosteric MALT1 inhibition via gliptins and its growth inhibitory effect on brain cancer cells.

[0857] Example 13: Sitagliptin inhibits the proteolvtic activity of MALT1

[0858] Inhibition of MALT1 proteolytic activity by sitagliptin is assessed. As positive controls, two MALT1 inhibitors were included, namely MI-2 and (S)-mepazine. Each of the compound is used at a concentration of 10 pM.

[0859] MI-2 (C19H17CI3N4O3; 2-chloro-N-[4-[5-(3,4-dichlorophenyl)-3-(2-methoxyethoxy)-1 ,2,4-triazol-1-yl] phenyl] acetamide) is an orthosteric and irreversible MALT1 inhibitor (Fontan et al, 2012).

[0860] (S)-mepazine (C19H22N2S; 10-{[(3S)-1 -Methyl-3-piperidinyl] methyl}-10H-phenothiazine) is an allosteric MALT1 inhibitor with higher binding affinity and inhibitory activity than (R)-mepazine or (±)-mepazine (Schlauderer et al, 2013).

[0861] A so-called buffer only test-condition (i.e. MALT1 + Ac-LRSR-AMC substrate + buffer) served as negative control.

[0862] The MALT1 activity is measured according to the activity assay developed by Fontan et al., 2012 where the MALT1 substrate peptide LRSR is linked to the fluorogen AMC (7-amino-4-methylcoumarin). Cleavage of the Ac-LRSR-AMC substrate by MALT1 results in release of AMC and a fluorescent signal. The recombinant full-length human MALT1 and Ac-LRSR-AMC - with excitation / emission wavelengths of 360 / 460 nm in black, low binding microtiter plates with a microplate reader are used for monitoring fluorogenic reactions. To eliminate false positives due to compound autofluorescence, two time points were measured for each reaction and hence, the fluorescence difference between such time points (T2-T1) was defined as MALT1 activity.

[0863] The final percent inhibition was calculated with the formula:

[0864] [fluorescence_test compound (T2-T1) - fluorescence_negative control (T2-T1)]

[0865] x 100 [fluorescence_positive control (T2-T1) - fluorescence_negative

[0866]

[0867] :-T1)]

[0868] Sitagliptin was identified as a MALT1 inhibitor using 40% inhibition as a threshold and validated against MI-2 and / or (S)-mepazine.

[0869] EC50 (effective concentration 50%) refers to the concentration of a test-compound at which 50% of its maximum effect is achieved. The term is quite general, regardless of whether a test-compound enhances or diminishes the biological parameter in question. Its equation is a modified version of theICso equation, taking the ratio of the response (R) to the maximum response (Rmax): {R / Rmax} = {1 / [1 +(E50 / l)n]}. In cases where a test-compound completely inhibits a biological activity at high dose, the values of EC50 and IC50 are identical (where Rmax = 100%). However, some test-compounds achieve only partial attenuation of a biological activity, even at high concentrations. In this case, IC50 values may be misleading about potency. Furthermore, efficacious test-compounds have more similar IC50 and EC50 values than their less efficacious counterparts. Of note and in contrast to IC50 values, EC50 values report on the binding affinity of the test-compound (effective concentration 50% is achieved when half of the target protein is bound). Using EC50 values allows the dose-response of test-compounds to be best quantified and reported when complex biological systems are employed, such as cell or animal models. Cell lines were in log phase of growth, harvested and seeded in a 96-well plate at optimum plating density. Next, cells were incubated with and without test-compounds (0.1 pM - 1 pM - 10 pM) in the presence of positive, negative and untreated controls. EC50 values were determined by Trypan blue dye-exclusion at t=48h and the IC50 calculator (Quest Graph™ IC50 Calculator." AAT Bioquest, Inc., 25 Apr. 2023, https: / / www.aatbio.com / tools / ic50-calculator).

[0870] Sitagliptin inhibits the proteolytic activity of MALT1 with an EC50 value of 1 .3 pM (Table 1 here below). Results are mean ± standard deviation of three independent experiments done in triplicates.

[0871] Table 1. Sitagliptin, MI-2 and (S)-mepazine EC50 values

[0872]

[0873] Materials and methods:

[0874] Cell Culture

[0875] To measure the MALT1 -dependent antiproliferative effect of sitagliptin on glioblastoma multiforme (GBM) cell growth and at the same time account for biases, a number of established and characterized GBM cell lines derived from GBM patient surgical samples was chosen (www.hgcc.se).

[0876] U3024 cell line is of mesenchymal subtype (human female, 73 years old) and is characterized by MALT 1 overexpression and very low DPP4 expression. U3005 is a cell line of proneural subtype (human male, 65 years of age) and is characterized by very low MALT1 expression and absent DPP4 expression. U3028 is of classical subtype (human female, 72 years of age) and is characterized by the absence of MALT1 expression and by DPP4 overexpression. In numbers:

[0877] U3024 cell line: MALT-1 expression 1.38 =; DPP4 expression = 0.15 (F, age of 73, subtype MESENCHYMAL); U3005 cell line: MALT-1 expression = 0.02; DPP4 expression = -0.63 (M, age of 65 years, subtype PRONEURAL); U3028 cell line: MALT-1 expression = -1.46; DPP4 expression = 1.66 (F, age of 72 years, subtype CLASSICAL). Schematically:

[0878] U3024 cell line: MALT-1 expression ++ / DPP4 expression -U3005 cell line: MALT-1 expression - I DPP4 expression - U3028 cell line: MALT-1 expression - / DPP4 expression ++

[0879] Cell lines were maintained in serum-free neural stem cell media to retain tumour-initiating capacity as well as tumour specific phenotypes (Pollard et al, 2009). The U87MG glioma cell line served as a bona fide human glioblastoma cell line. Cell cultures were performed in 3D.

[0880] Example 14 - Cell viability and cytotoxicity assay

[0881] To assess cell viability and cytotoxicity, the Calcein AM-EthD III Vi ability / Cytotoxicity Assay (30002, Biotium, San Francisco, United States) was performed on U3005-MG spheroids grown from U3005 cells, U3024-MG spheroids grown from U3024 cells, and U3028-MG spheroids grown from U3028 cells, treated with test compounds. Seven concentrations of the test compounds (0.001 pM, 0.01 pM, 0.1 pM, 5.0 pM, 10 pM, 50.0 pM, and 100 pM) were evaluated in a dose-response manner. For staining, dye stock solutions of Calcein AM (2 pM final concentration) and Ethidium homodimer III (EthD III, 4 pM final concentration) were first warmed to room temperature (RT) and mixed by vortexing for 10 seconds to ensure homogeneity. The staining solution was prepared in PBS (1 x, without calcium and magnesium) and vortexed for 15 seconds for thorough mixing. Spheroids were collected by centrifugation at 1000 x rpm for 5 minutes at RT, and the supernatant was carefully removed. Each spheroid pellet was resuspended in 100 pL of the staining solution, ensuring minimal disruption to spheroid integrity. The stained spheroids were incubated at RT for 30 to 45 minutes under sterile conditions. Imaging of the stained spheroids was conducted using the Lionheart FX Automated Microscope (BioTek, Winooski, United States). Calcein fluorescence, indicative of live cells, was detected using a GFP filter, while EthD III fluorescence, marking dead cells, was captured using a Texas Red filter. Maximum projection images were generated from Z-stacks of seven images, each taken 30 pm apart, to provide detailed three-dimensional visualization. Quantitative analysis of spheroid properties, including size, area, and circularity, was performed using Gen5 software, version 3.14 (BioTek, Winooski, United States). Spheroid size (pm) was determined by fitting an ellipse to the spheroid contour, while the area (pm2) was calculated by converting pixel counts to actual area measurements based on the pixel dimensions. Circularity was also calculated to provide insights into spheroid morphology. Cytotoxicity was quantified using the following formula: Cytotoxicity (%)= (lnt[EthDIII]) / (lnt[EthDIII]+lnt[Calcein]) x 100, where lnt[EthD III] and lnt[Calcein] represent the fluorescence intensities of EthD III and Calcein, respectively. This comprehensive methodology ensures reliable assessment of cell viability and cytotoxicity in spheroids under various treatment conditions. The Positive control for cell killing: Dead cell controls were prepared by treating cells with 0.1% saponin or 0.5% digitonin for 10 minutes. Cells were also killed by heating to 56°C for 45 minutes, and then cooled to room temperature.

[0882] Results are depicted in Table A2 here below.

[0883] Table A2. The effects of S-mepazine and gliptins on spheroids of patient-derived glioma cells sharing various levels of MALT-1 expression. Three independent experiments done in triplicates. Sitagliptin and positive control and (S)-mepazine, were tested at 0.001-100 pM. DMSO was <0.05%.

[0884] > >

[0885] >

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[0888]

[0889] It is concluded that gliptins, and particularly sitagliptin decreases patient-derived glioma cell metabolism.

[0890] Example 15: Cell proliferation assayAs positive controls, two MALT1 inhibitors were included, namely MI-2 and (S)-mepazine. MI-2 (C19H17CI3N4O3; 2-chloro-N-[4-[5-(3,4-dichlorophenyl)-3-(2- methoxyethoxy)-1 ,2,4-triazol-1 -y I] phenyl] acetamide) is an orthosteric and irreversible MALT1 inhibitor (Fontan et al, 2012). (S)-mepazine (C19H22N2S; 10-{[(3S)-1-Methyl-3- piperidinyl] methyl}-10H-phenothiazine) is an allosteric MALT1 inhibitor with higher binding affinity and inhibitory activity than (R)-mepazine or (±)-mepazine (Schlauderer et al., 2013).

[0891] Cell lines were in log phase of growth, harvested and seeded in a 96-well plate at optimum plating density. Next, sitagliptin and positive controls, MI-2 and (S)-mepazine were tested at 0.1 pM, 1 pM, and 10 pM at t=48h. DMSO was <0.05%.

[0892] Figure 11C shows the antiproliferative effects of MI-2, mepazine and sitagliptin on U87-MG cell growth. The results are mean ± standard deviation of three independent experiments done in triplicates.

[0893] Figure 12A shows the inhibition of MALT1 by sitagliptin at 10 pM in patient-derived glioblastoma cells (U3024) that over-express MALT1 resulting in spheroid disintegration (upper pane) when compared to untreated spheroids (lower pane).

[0894] Figure 12B shows the inhibition of MALT 1 by MI-2, mepazine and sitagliptin in patient-derived glioblastoma cells (U3024). Three independent experiments were performed in triplicates.

[0895] The above results clearly show that sitagliptin exhibits strong antiproliferative effect on GBM cell growth.

[0896] Example 16: Proteolytic inhibition of MALT1

[0897] Sitagliptin inhibition of MALT1 proteolytic activity was performed at 10 pM. Fluorogenic reactions were monitored in the presence of recombinant full-length human MALT1 and Ac LRSR-AMC with excitation / emission wavelengths of 360 / 460 nm in black, low binding microtiter plates using a microplate reader. Cleavage of the Ac-LRSR-AMC substrate by MALT1 results in the release of AMC and a fluorescent signal. To eliminate false positives due to compound autofluorescence, two time points were measured for each reaction and hence, the fluorescence difference between such time points (T2-T1) was defined as MALT1 activity. The final percent inhibition was calculated with the formula: {[fluorescence_test compound(T2-T1) -fluorescence_negative control (T2-T1)] / [fluorescence_positive control (T2-T1) - fluorescence_negative control (T2-T1)]} x 100. (S)-mepazine or MI-2 (C19H17CI3N4O3; 2 chloro-N-[4-[5-(3,4-dichlorophenyl)-3-(2-methoxyethoxy)-1 ,2,4-triazol-1-yl] phenyl] acetamide) were used as positive controls at 10 pM. A so-called buffer only test-condition (i.e. MALT1+Ac-LRSR-AMC substrate + buffer) served as negative control. Sitagliptin was identified as a MALT1 inhibitor using 40% inhibition as a threshold and validated against MI-2 and / or (S)-mepazine.

[0898] EC50 (effective concentration 50%) refers to the concentration of a test-compound at which 50% of its maximum effect is achieved. The term is quite general, regardless of whether a test-compound enhances or diminishes the biological parameter in question. Its equation is a modified version of the IC50 equation, taking the ratio of the response (R) to the maximum response (Rmax): {R / Rmax} = {1 / [1 +(E50 / l)n]}. In cases where a test-compound completely inhibits a biological activity at high dose, the values of EC50 and IC50 are identical (where Rmax = 100%). However, some test-compounds achieve only partial attenuation of a biological activity, even at high concentrations. In this case, IC50 values maybe misleading about potency. Furthermore, efficacious test-compounds have more similar IC50 and EC50 values than their less efficacious counterparts. Of note and in contrast to IC50 values, EC50 values report on the binding affinity of the test-compound (effective concentration 50% is achieved when half of the target protein is bound). Using EC50 values allows the dose response of test-compounds to be best quantified and reported when complex biological systems are employed, such as cell or animal models.

[0899] Cell lines were in log phase of growth, harvested and seeded in a 96-well plate at optimum plating density. Next, cells were incubated with and without test-compounds (0.1 pM - 1 pM - 10 pM) in the presence of positive, negative and untreated controls. EC50 values were determined by Trypan blue dye-exclusion at t=48h and the IC50 calculator (Quest Graph™ IC50 Calculator." AAT Bioquest, Inc., 25 Apr. 2023, www.aatbio.com / tools / ic50 calculator).

[0900] Sitagliptin inhibits the proteolytic activity of MALT1 with an EC50 value of 1.3 pM (Table A3 here below). Results are mean ± standard deviation of three independent experiments done in triplicates.

[0901] Table A3: Sitagliptin, MI-2 and (S)-mepazine EC50 values

[0902] Test-compounds EC50 values (pM)

[0903] Sitagliptin 1.3

[0904] MI-2 1.0

[0905] (S)-mepazine 1.0

[0906] Example 17: Enzvme (catalytic) activity of the CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 isoenzymes

[0907] CYP450s are the major mediators of xenobiotics’ toxicity and interactions. Ideally, someone would like to have xenobiotics that do not induce or inhibit those enzymes. In a nutshell, here presented data show that these enzymes function properly and unaltered (metabolic and catalytic activities) under influence of exposure to gliptins and when inhibition is considered, sitagliptin has the most favourable profile among gliptins but also when compared to mepazine. CYP2D6 and CYP3A4 inhibition is the most detrimental effect of mepazine as those enzymes are involved in most xenobiotics’ metabolism to date. The enzymes selected for testing of influence of gliptins and mepazine are the seven most relevant enzymes of the kind in humans.

[0908] To determine the isozyme-specific CYP450-metabolism, baculosomes® expressing CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 human CYP450 isoenzymes were purchased from Thermo Fisher Scientific (Waltham, MA, USA). All reagents were handled and prepared according to the manufacturer’s protocol. Sitagliptin was tested at 1 pM. Enzymatic activity in the presence of sitagliptin was assessed based on the kinetic model for CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 isoforms. Reactions took place in triplicates at 20°C. Glucose-6-phosphate at 333 mM and glucose-6-phosphate-dehydrogenase at 30 U / mL in 100 mM potassium phosphate, pH 8.0 (regeneration system) converted NADP+ (10 mM in 100 mM potassium phosphate, pH 8.0) into NADPH to initiate the CYP450 reaction. Upon the addition of the fluorescentsubstrate, immediately (in the presence of test-compound; A is the rate observed in the presence of negative (solvent, DMSO) control.

[0909] The administration of test-compounds at 1 pM (60 minutes) and Relative Fluorescence Units (RFU) measured for the reaction under different conditions.

[0910] CYP450 (%) inhibition of CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 isoenzymes were measured upon the administration of test compounds at 1 pM (60 minutes).

[0911] Data show that sitagliptin, saxagliptin, alogliptin and linagliptin do not inhibit nor induces CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 human CYP450 isoenzymes.

[0912] It was apparent that mepazine showed substantial influence on CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 isoenzymes, specifically on CYP2D6.

[0913] Example 18 - cell metabolism under influence of qliptins

[0914] (S)-mepazine and sitagliptin both reduced cell metabolism in a dose-dependent manner.

[0915] Figure 13A-C show the percentage of cell metabolism relative to untreated controls after treatment with (S)-mepazine (positive control) and sitagliptin at various concentrations (1 pM, 10 pM, and 50 pM). Cell metabolism was assessed using the Alamar Blue assay. Error bars represent mean ± standard deviation from triplicate experiments.

[0916] Overall, based on the insights gained with the examples 1-6, it is concluded that gliptins are MALT1 inhibitors capable of killing tumor cells, as determined with computational exploration of binding capabilities of the gliptins to the allosteric binding site and as determined with confirmative in vitro cellbased assays using human cancer-patient derived glioma cells. Thus, the experimental data align with the computational data. Gliptins identified as allosteric MALT1 inhibitors based on computational data and based on in vitro cell-based assay data match.

[0917] REFERENCES relating to (prophylactic) treatment of allergic inflammatory disease

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[0926] • Kim N-H, Yu T, Lee DH. Biomed Res Int 2014, 2014: 368703.• Kushwaha, Ram & Haq, Wahajul & Katti, Seturam. (2014). Discovery of 17 Gliptins in 17-Years of Research for the Treatment of Type 2 Diabetes: A Synthetic Overview. Chemistry & Biology Interface. 4. 137-162.

[0927] • Liang X, Cao YC et al 2021 , Med Res Rev 41 (4):2388-2422.

[0928] • Liu D, Jin B, Chen W, Yun P. BMC Pharmacol Toxicol 2019, 20(1): 15.

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[0930] "Paracaspase MALT1 deficiency protects mice from autoimmune mediated demyelination." J Immunol 190(6): 2896-2903.

[0931] • McNutt, Andrew T., et al. "GNINA 1.0: molecular docking with deep learning." Journal of cheminformatics 13.1 (2021): 43.

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[0934] • Quancard, Jean, et al. "An allosteric MALT1 inhibitor is a molecular corrector rescuing function in an immunodeficient patient." Nature chemical biology 15.3 (2019): 304-313.

[0935] • Sakura H, Hashimoto N, Sasamoto K, Ohashi H, Hasumi S et al. BMC Endocr Disord 2016, 16(1):

[0936] • Sarwer-Foner, G.J., and Koranyi, E.K. (1957). The clinical investigation of pacatai in open psychiatric settings. Can. Med. Assoc. J. 77, 450-459.

[0937] • Schlauderer F, Structural and functional analysis of MALT1 inhibition by phenothiazine derivatives, Dissertation zur Erlangung des Doktorgrades der Fakultat fur Chemie und Pharmazie der Ludwig-Maximilians-Universitat Munchen, 2015

[0938] • Schlauderer F, Lammens K, Nagel D, Vincendeau M, Eitelhuber AC et al. Angew Chem Int Ed Engl 2013, 52(39): 10384.

[0939] • Tran, T.D, et al., “Secondary Metabolites from the Fungus Dictyosporium sp. and Their MALT1 Inhibitory Activities” DOI: 10.1021 / acs.jnatprod.8b00871 , J. Nat. Prod. 2019, 82, 154-162. • Wang, Pathogenesis of allergic diseases and implications for therapeutic interventions, Signal Transduction and Targeted Therapy (2023) 8:138

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[0942] REFERENCES relating to (prophylactic) treatment of autoimmune disease, preferably MS

[0943] 1. Filipp...

Claims

1. CLAIMS1. A pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof,for use in a therapeutic or prophylactic method of treating a MALT-1 mediated disease selected from:allergic inflammatory disease,an autoimmune disease, preferably multiple sclerosis,multiple myeloma, anda neurological cancer, preferably glioma,in a subject, preferably a human subject, in need thereof, orfor use in a therapeutic method of treating neurological cancer in a human subject in need of the treatment, wherein the neurological cancer is positive for expression of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1).

2. The pharmaceutical composition for use according to claim 1 , wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin or cetagliptin, preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, more preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin or alogliptin, most preferably wherein the gliptin is sitagliptin.

3. The pharmaceutical composition for use according to claim 1 , wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, or cetagliptin, preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin or alogliptin, more preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin or alogliptin.

4. The pharmaceutical composition for use according to claim 1 , wherein the gliptin is not sitagliptin.

5. The pharmaceutical composition for use according to claim 1 , wherein the gliptin is an inhibitor of mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1), preferably an allosteric inhibitor of MALT1.

6. Pharmaceutical composition for use according to any one of the claims 1-5, wherein the therapeutic effect of the gliptin comprises and / or results from inhibition of MALT1 , preferably wherein the inhibition of MALT1 is an allosteric inhibition resulting from binding of the gliptin to an allosteric site within MALT1.

7. Pharmaceutical composition for use according to any one of the preceding claims 1 and 5-6, wherein the neurological cancer is negative for expression of dipeptidyl peptidase-4 (DPP-4) or comprises DPP-4 expression considered as unchanged to or lower as compared to a healthy reference tissue, preferably being the tissue from which the cancer originated and / or non-cancerous tissue surrounding the cells of the cancer, advantageously being a healthy tissue of the subject.

8. Pharmaceutical composition for use according to any one of the preceding claims 1 and 5-7, wherein MALT1 expression in the neurological cancer is assessed as MALT1 overexpression.

9. Pharmaceutical composition for use according to any one of the claims 1 and 5-8, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin;preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin;more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin, and alogliptin, most preferably wherein the gliptin is sitagliptin.

10. Pharmaceutical composition for use according to any one of the claims 1 and 5-8, wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, and cetagliptin;preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin;more preferably wherein the gliptin is of vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin;even more preferably of linagliptin, vildagliptin, saxagliptin and alogliptin,most preferably wherein the gliptin is linagliptin.

11. The pharmaceutical composition for use according to any one of the preceding claims, wherein the method comprises oral administration of the pharmaceutical composition.

12. The pharmaceutical composition for use according to any one of the preceding claims 1-6 and 11 , wherein the method results in inhibition of intracellular MALT1 activity, preferably inhibition of MALT1 activity inside aberrant cells, preferably aberrant Th2 cells or preferably aberrant Th1 cells and / or Th17 cells, or preferably inhibition of MALT1 activity inside myeloma cells, or inhibition of MALT1 -mediated activation of NF-KB signaling pathway, or inhibition of cytokines regulated by MALT1 -mediated NF-KB signaling, such as TNF-a and / or IL-6.

13. The pharmaceutical composition for use according to any one of the preceding claims 1-6 and 11- 12, wherein:the allergic inflammatory disease is a chronic allergic condition and / or a late-phase allergic condition;the MS is clinically isolated syndrome (CIS), relapsing-remitting MS (RRMS), secondary progressive MS (SPMS), primary progressive MS (PPMS) or progressive relapsing MS (PRMS).

14. The pharmaceutical composition for use according to any one of the preceding claims 1-6 and 11- 13, wherein:the allergic inflammatory disease is selected from the group consisting of allergic asthma, anaphylaxis, sinusitis, allergic rhinitis, eczema, hives, food allergy, chronic urticaria, atopic dermatitis and ocular allergic diseases;the MS is at pre-symptomatic stage, clinically isolated syndrome stage, relapsing-remitting stage, or secondary progressive stage.

15. The pharmaceutical composition for use according to any one of the preceding claims, wherein the MALT-1 mediated disease selected from allergic inflammatory disease, an autoimmune disease, preferably multiple sclerosis, multiple myeloma and a neurological cancer such as glioma is mediated by or dependent on MALT1 activity.

16. The pharmaceutical composition for use according to any one of the preceding claims, wherein the method comprises the step of diagnosing a subject as suffering from the MALT-1 mediated disease selected from allergic inflammatory disease, an autoimmune disease, preferably multiple sclerosis, multiple myeloma and a neurological cancer such as glioma, or being at risk of suffering from the MALT-1 mediated disease selected from allergic inflammatory disease, an autoimmune disease, preferably multiple sclerosis, multiple myeloma and a neurological cancer such as glioma, mediated by or dependent on MALT1 activity.

17. Pharmaceutical composition for use according to any one of the claims 1 , 5-11 and 15-16, wherein the neurological cancer is brain cancer and / or is glioma, neuroblastoma, or medulloblastoma; preferably being glioma or neuroblastoma, more preferably being glioblastoma or gliosarcoma, even more preferably being malignant glioblastoma, most preferably being glioblastoma multiforme.

18. Pharmaceutical composition for use according to any one of the claims 1 , 5-11 and 15-17, wherein the treatment comprises administering to the human subject of an effective amount of the compound, preferably directly into the neurological cancer or into the cavity remaining after surgical removal of the neurological cancer, possibly in combination with one or more chemotherapeutic agent and / or anticancer agent specific to the cancer of the subject.11919. Pharmaceutical composition for use according to any one of the preceding claims, wherein the treatment is followed by radiotherapy and / or chemotherapy.

20. Pharmaceutical composition for use according to any one of the preceding claims, wherein MALT1 expression status, and optionally DPP-4 expression status, comprises RNA expression analysis and / or protein expression analysis, preferably by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and / or immunoassay.

21. The pharmaceutical composition for use according to any one of the preceding claims, wherein the method comprises the oral administration of the gliptin, pharmaceutically acceptable salt thereof or hydrate thereof, at a dose of 0.01 mg to 1000 mg per day.

22. The pharmaceutical composition for use according to any one of the preceding claims, wherein the pharmaceutical composition is provided in an oral unit dosage form, preferably in the form of a tablet, comprising the gliptin, the pharmaceutically acceptable salt thereof or hydrate thereof, in an amount of 0.01 mg to 1000 mg.

23. A therapeutic or prophylactic method of treating a MALT-1 mediated disease selected from allergic inflammatory disease, multiple sclerosis, multiple myeloma and glioma, in a subject, preferably human subject, in need thereof, said method comprising the step of administration to said subject of a pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof.

24. Use of a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, in the manufacture of a medicament for the therapeutic or prophylactic treatment of a MALT-1 mediated disease selected from allergic inflammatory disease, multiple sclerosis, multiple myeloma and glioma, in a subject, preferably human subject, in need thereof.

25. An in vitro method for determining an increased probability that a neurological cancer in a human subject will respond to the treatment witha pharmaceutical composition comprising a compound selected from the group consisting of gliptins, pharmaceutically acceptable salts thereof and hydrates thereof, wherein the gliptin is selected from wherein the gliptin is selected from any one or more of sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, , cofrogliptin, fotagliptin, prusogliptin, dutogliptin, neogliptin, and cetagliptin; the method comprising the steps of:- providing a test first sample obtained from the subject;- detecting the presence of MALT1 expression and preferably assessing the expression level of MALT 1 in the first test sample;120- preferably, comparing the assessed expression level of MALT1 to a reference MALT1 expression level;wherein finding that MALT1 expression is present in the first test sample and preferably that the expression level of MALT1 assessed in the first test sample is higher as compared to the reference MALT 1 expression level is indicative of an increased probability that the subject will respond to the treatment.

26. The in vitro method according to claim 25, further comprising the steps of:- providing a second test sample obtained from the subject;- detecting the presence of DPP-4 expression or optionally assessing the expression level of DPP-4 or DPP-4 enzymatic activity level in the second test sample;- preferably, comparing the assessed expression level or enzymatic activity level of DPP-4 to a reference DPP-4 expression level or a reference DPP-4 enzymatic activity level, respectively; wherein finding that DPP-4 expression is absent in the second test sample or optionally that the expression level of DPP-4 or DPP-4 enzymatic activity level assessed in the second test sample is lower as compared to the reference DPP-4 expression level or a reference DPP-4 enzymatic activity level, respectively; preferably finding that DPP-4 expression is absent in the second test sample is indicative of an increased probability that the subject will respond to the treatment, wherein the response is further associated with an increased probability of not causing adverse DPP-4-inhibition-associated side effects.

27. The in vitro method according to any one of the claims 25-26, wherein the first test sample and / or the second test sample is or comprises biopsy sample, cerebrospinal fluid (CSF) sample, or a blood sample, preferably being biopsy sample or the cerebrospinal fluid (CSF) sample, even more preferably being biopsy sample.

28. The in vitro method according to any one of the claims 25-27, wherein detecting the presence of expression and / or assessing the expression level comprises RNA analysis, preferably comprising RNA-sequencing or reverse transcription-quantitative polymerase chain reaction (RT-qPCR), more preferably comprising RT-qPCR.

29. The in vitro method according to any one of the claims 25-28, wherein detecting the presence of expression and / or assessing the expression level comprises protein analysis, preferably comprising an immunoassay, more preferably comprising ELISA.

30. The in vitro method according to any one of the claims 26-29, wherein the first test sample and the second test sample are the same test sample, preferably wherein detecting the presence of MALT1 and DPP-4 expression, optionally assessing the expression level of MALT1 and DPP-4 is performed as part of the same assay, more preferably being a multiplex RT-qPCR assay or an immunoassay.12131. The in vitro method according to any one of the claims 25-30, wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin; preferably wherein the gliptin is sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin;more preferably wherein the gliptin is sitagliptin, linagliptin, vildagliptin, saxagliptin, and alogliptin, most preferably wherein the gliptin is sitagliptin;and / orwherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, dutogliptin, neogliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, and cetagliptin;preferably wherein the gliptin is vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, alogliptin, teneligliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, and prusogliptin;more preferably wherein the gliptin is of vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, and alogliptin;even more preferably of linagliptin, vildagliptin, saxagliptin and alogliptin,most preferably wherein the gliptin is linagliptin.

32. The in vitro method according to any one of the claims 25-31 , wherein the reference MALT1 expression value, and optionally the reference DPP-4 expression value, is defined and has (have) been predetermined based on assessing MALT1 expression levels, and optionally DPP-4 expression levels, respectively, in a plurality of control samples obtained from healthy control subjects.

33. The in vitro method according to any one of the claims 25-31 , wherein the reference MALT1 expression value, and optionally the reference DPP-4 expression value, is determined based on assessing normal MALT1 expression levels, and optionally normal DPP-4 expression levels, respectively, in a healthy tissue sample obtained from the subject, preferably being a healthy tissue corresponding to the tissue from which the cancer originated, optionally being healthy non-cancerous tissue surrounding the cancer cells.

34. The in vitro method according to any one of the one of the claims 25-32, wherein the expression level of MALT1 assessed for the first test sample is at least 2 fold higher than the reference MALT1 expression value, preferably being at least 5 fold higher; more preferably being at least 10 fold higher.122

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