Triazolo compounds for use in the treatment of graft-versus-host disease

Abstract

The present invention relates to a compound of Formula (I), in particular, the compound of Formula (II) or an isotopic variant, a pharmaceutically acceptable salt or solvate thereof for use in a method of treatment of graft-versus-host disease in a subject in need thereof.

Description

[0001] TRIAZOLO COMPOUNDS FOR USE IN THE TREATMENT OF GRAFT-VERSUS-HOST DISEASE

[0002] SUMMARY OF THE INVENTION

[0003] The present invention relates to a compound of Formula (I), in particular, a compound of Formula (II) or an isotopic variant, a pharmaceutically acceptable salt or solvate thereof for use in a method of prevention or treatment of graft-versus-host disease (GvHD) in a subject in need thereof.

[0004] BACKGROUND

[0005] Graft-versus-host disease (GvHD) is a significant and potentially life-threatening complication that can occur after allogeneic hematopoietic stem cell transplantation (HSCT). In this condition, the donated bone marrow or peripheral blood stem cells view the recipient's body as foreign and mount an immune response against the host's tissues. GvHD is broadly categorized into two types: acute GvHD, which typically manifests within the first 100 days post-transplant, and chronic GvHD, which can develop anytime thereafter. The severity of GvHD can range from mild to severe, impacting various organs and systems, including the skin, liver, gastrointestinal tract, and lungs.

[0006] The pathophysiology of GvHD involves complex interactions between donor T cells and host antigen-presenting cells. Initially, donor T cells are activated by recognizing host antigens as foreign. This activation triggers a cascade of inflammatory responses, leading to tissue damage and the clinical manifestations of GvHD. Acute GvHD is often characterized by skin rashes, liver dysfunction, and gastrointestinal symptoms such as diarrhea and abdominal pain. Chronic GvHD, on the other hand, resembles autoimmune disorders and can cause fibrosis and sclerosis of various tissues, leading to long-term morbidity.

[0007] Current treatment strategies for GvHD primarily involve immunosuppressive therapies aimed at reducing the activity of donor immune cells. First-line treatment typically includes corticosteroids, which can effectively manage symptoms in many patients. However, a significant proportion of patients are refractory to steroids or develop severe side effects, necessitating the use of additional immunosuppressive agents such as JAK2 inhibitors, calcineurin inhibitors, mycophenolate mofetil, and monoclonal antibodies. Despite these treatments, GvHD remains a major cause of non-relapse mortality and morbidity following HSCT, highlighting the need for novel therapeutic approaches.

[0008] A conditioning regimen used to prepare a patient for stem cell transplantation may include chemotherapy, monoclonal antibody therapy and / or radiation to the entire body. This conditioning is crucial to eradicate underlying disease and to support engraftment of donor cells without rejection by recipient. However, conditioning-mediated damage to the gastrointestinal tract and reduced function of epithelial barrier allows systemic translocation of microbial products such as lipopolysaccharide and other pathogen-associated molecular patterns that greatly amplify host immune system activation. For that reason, the most common histopathological finding in acute GvHD is gastrointestinal epithelial cell apoptosis (Nat. Rev. Gastroenterol. Hepatol. 2017;14:711). Intestinal stem cells are targets of the disease and the most prominent histological manifestation is apoptotic bodies in the regenerative compartment of the crypt (Hum. Pathol. 2009;40:909). In addition, the number of paneth cells, which are primarily located next to stem cells in small intestinal crypts, inversely correlates with the risk of mortality in acute GvHD.

[0009] Addressing the tissue damage that occurs is of paramount importance. While immunosuppressive therapy plays a crucial role, a multifaceted approach that encompasses both the mitigation of tissue damage, strengthening the epithelial barrier in the gut and the modulation of the host's biological response is essential for achieving optimal therapeutic outcomes in patients undergoing allogenic HSCT.

[0010] DETAILED DESCRIPTION OF THE INVENTION

[0011] In one aspect, the present invention relates to a compound of Formula (I) Formula (I), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft -versus-host disease in a subject in need thereof, wherein:

[0012] R1is selected from the group consisting of hydrogen, carboxyl, cyano, fluorine, chlorine, methyl, isopropyl, t-butyl, trifluoromethyl, trifluoromethoxy, cyclopropylmethoxy, 1,1- difluoro-2-methylpropyl, l,l-difluoro-2,2-dimethylpropyl, 1-methyl-l-cyclobutyl, methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl, 1-hydroxy-l-methylethyl, azetidine- 1-carbonyl, 3-methyloxetan-3-yl, 4,5-dihydrooxazol-2-yl and cyclopropylcarbonyl;

[0013] R2is hydrogen or methyl;

[0014] A1is =N- or =CH-;

[0015] X is benzene, pyridine or cyclohexane;

[0016] J is any ring selected from the group

[0017]

[0018] Y is selected from the group consisting of phenyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, piperidinyl, azetidinyl, tetrahydropyranyl, morpholinyl and tetrahydropyridinyl, optionally substituted with 1, 2 or 3 groups selected from Y1, or alternatively Y is amino, optionally substituted with 1 or 2 groups selected from Y1;

[0019] Y1is selected from the group consisting of hydroxy, cyano, fluorine, amino, methyl, ethyl, isopropyl, methoxy, azetidinyl, pyrrolidinyl, morpholinyl, , wherein amino is optionally substituted with 1 or 2 groups selected from Y2and methyl, ethyl, isopropyl, methoxy, azetidinyl, pyrrolidinyl, morpholinyl, are optionally substituted with 1,

[0020] 2 or 3 groups selected from Y2;

[0021] Y2is selected from the group consisting of hydroxyl, fluorine, amino, methyl, methoxy, azetidinyl, pyrrolidinyl, morpholinyl, wherein amino is optionally substituted with 1 or 2 groups selected from Y3and methyl, methoxy, azetidinyl, pyrrolidinyl, morpholinyl, is optionally substituted with 1, 2 or 3 groups selected from Y3,

[0022] Y3is hydroxyl, fluorine, cyano, methyl or methoxy, wherein methyl or methoxy are optionally substituted with fluorine.

[0023] In some embodiments, the compound of Formula (I) is selected from the following group:

[0024] or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof.

[0025] In some embodiments, the compound of Formula (II) is or an isotopic variant, a pharmaceutically acceptable salt or solvate thereof.

[0026] In some embodiments, the compound of Formula (II) is

[0027] The compound of Formula (I), in particular the compound of Formula (II), targets sirtuin 6 (SIRT6), a member of a family of proteins related to the founding member of the family - the silent information regulator 2 protein (Sir2p) of Saccharomyces cerevisiae, a nicotinamide adenine dinucleotide (NAD+)-dependent histone deacetylase (HDAC) regulating chromatin silencing. Mammals contain at least seven sirtuin homologues numbered SIRT1 to SIRT7, characterised by significant sequence homology, particularly within their conserved NAD+- binding domains (Biochem. Biophys. Res. Commun. 2000;273:793). SIRT6 serves as a transcriptional regulator of intestinal barrier function and regeneration of bowel epithelium. Based on preclinical data, the compound represents a unique treatment approach, as the mechanism of action targets the restoration of the intestinal barrier function and bowel wall architecture in patients suffering from gastrointestinal diseases such as celiac disease, atypical wheat allergy, gluten intolerance (e.g., non-celiac gluten sensitivity (NCGS), gluten ataxia, dermatitis herpetiformis (DH) and wheat allergy), inflammatory bowel disease (IBD) (e.g., Crohn's disease, ulcerative colitis or IBD-associated diarrhea), irritable bowel syndrome (IBS), (especially irritable bowel syndrome with diarrhea (IBS-D)), microscopic colitis (e.g., collagenous colitis and lymphocytic colitis) and eosinophilic esophagitis (EoE).

[0028] Targeting SIRT6 with a compound of Formula (I), in particular the compound of Formula (II), prevents the deacetylation of one of its ligands, acetylated lysine (K) 56 of histone H3 (H3K56), in an in-vitro cell-free and cellular system and significantly accelerates the recovery of transepithelial electrical resistance (TEER) across a monolayer of colorectal adenocarcinoma- 2 (Caco-2) cells following inflammation with tumor necrosis factor (TNF-a) or interleukin (I L)- 6. In one TEER model, tight junction (TJ)-related proteins were investigated, and a normalisation of relevant TJ proteins (claudin (Cldn)-l and claudin-2) was observed. Interaction with a compound of Formula (I) is highly selective for SIRT6 over other members of the human SIRT protein family and does not inhibit enzymatic activity of other histone deacetylases (HDACs).

[0029] Compounds of Formula (I), in particular the compound of Formula (II), has shown to increase expression of SIRT6 protein in patient samples / biopsies, cell cultures and organoids. At the same time, an increase in H3K56 acylation in cell culture and organoids can be observed. Both effects have an influence on target gene regulation and therefore cannot be isolated. A consistent upregulation of protective gastrointestinal genes in intestinal tissue after treatment with a compounds of Formula (I), in particular the compound of Formula (II), compared to vehicle control can be observed, while in non-stimulated intestinal cells (Caco- 2) no change in gene regulation was observed after treatment. This implies, that compounds of Formula (I) have a positive effect on the regulation of important target genes, specifically in the disease situation, especially enhancing the natural regenerative process (regeneration of crypts, where intestinal stem cells are located which undergo asymmetric division, thereby producing fully differentiated epithelial gut cells and renewing intestinal stem cells. These new epithelial cells renew the lining of crypts and villi and maintain / restore a healthy gut and proper intestinal barrier) in the gut, which may already been damaged in the conditioning phase during GvHD treatment.

[0030] Several treatments exist to ameliorate the conditioning phase in GvHD. These treatments are designed to reduce the intensity of the conditioning regimen and minimize its adverse effects while still preparing the patient for HSCT. The main approaches include: (a) reduced-intensity conditioning (RIC) regimens: RIC regimens use lower doses of chemotherapy and radiation compared to traditional myeloablative conditioning. This approach aims to reduce toxicity while still allowing for effective engraftment of donor cells. Common agents used in RIC include: fludarabine, nelphalan and busulfan (at reduced doses); (b) non-myeloablative conditioning: these regimens use very low doses of chemotherapy and radiation, focusing more on immunosuppression rather than myeloablation. This approach is particularly beneficial forolder patients orthose with comorbidities. Common agents include: fludarabine or low-dose total body irradiation; (c) immunosuppressive therapies: certain therapies can be used to reduce the toxicity of the conditioning regimen, e.g. monoclonal antibodies such as alemtuzumab or rituximab, which can be used to deplete specific immune cells, reducing the risk of GvHD; (d) immunosuppressive medications, used to prevent GvHD after transplantation can also be part of the conditioning strategy, e.g. such as calcineurin inhibitors, methotrexate or mycophenolate mofetil; (e) experimental approaches like selective ROCK2 inhibitors (e.g. belumosudil) that modulate immune responses or gene therapy which modifies donor cells to reduce the risk of GvHD; and (f) protective agents and supportive care: these are used to protect organs and mitigate side effects during conditioning like amifostine (a cytoprotective agent used to reduce the harmful effects of chemotherapy and radiation), mesna (used to protect the bladder from the harmful effects of certain chemotherapy drugs) and growth factors (such as granulocyte colony-stimulating factor) to promote bone marrow recovery.

[0031] The goal is to achieve effective engraftment of donor cells while minimizing the risk and severity of GvHD. And for this purpose, selective SIRT6 modulator such as compounds of Formula (I), in particular the compound of Formula (II), can also be used which result in a positive clinical effect on maintenance of gut epithelium, recovery of the gut from damage by chemotherapy and radiotherapy, diversity and composition of gut microbiome, translocation of microbes and microbial products, and can also have a beneficial therapeutic effect against immune activation. In this seting, a compound of Formula (I), in particular the compound of Formula (II), has a protective effect against GvHD after allogeneic HSCT that can be used for therapeutic benefit in humans, leading to decreased rates of GvHD after transplant, decreased use of immunosuppressive agents after transplant, decreased rate of infections by gut bacteria after transplant and improved survival of transplant patients.

[0032] In summary, compounds of Formula (I), in particular the compound of Formula (II), is a highly selective and very potent biochemical inhibitor of the deacetylase and ribosyltransferase activity of SIRT6 and at the same time induces stability and enhanced expression of SIRT6 protein due to the unique irreversible binding mode called "ribosylation trapping" in the substrate binding site of the triazolo moiety in Formula (I).

[0033] In some embodiments, said compound of Formula (I), in particular the compound of Formula (II), is capable of restoring intestinal barrier function, inducing physiologic regeneration of the crypt cells.

[0034] In some embodiments, said subject is human. In some embodiments, said subject suffers from a disease selected from chronic GvHD (cGvHD), acute GvHD (aGvHD), gastrointestinal GvHD, steroid-refractory GvHD, steroid- resistant GvHD or transfusion-associated graft-versus-host disease.

[0035] In some embodiments, said subject suffers from suffers from gastrointestinal GvHD.

[0036] In some embodiments, said subject suffers from suffers from steroid-refractory GvHD or steroid-resistant GvHD.

[0037] In some embodiments, said subject is at risk of developing aGvHD.

[0038] In some embodiments, said subject suffers from a disease selected from malignant disorders (such as acute myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, myelodysplastic syndromes, myeloproliferative disorders, non-Hodgkin's lymphoma, Hodgkin's disease, chronic lymphocytic leukemia, multiple myeloma, juvenile chronic myeloid leukemia), non-malignant disorders (such as aplastic anemia, paroxysmal nocturnal hemoglobinuria, Fanconi's anemia, Blackfan-Diamond anemia, sickle cell anemia, thalassemia major, severe combined immunodeficiency, Wiskott-Aldrich syndrome, inborn errors of metabolism) or a planned conditioning regimen in the context of an allogeneic HSCT.

[0039] In some embodiments, said subject suffers from a malignant disorders selected from acute myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, myelodysplastic syndromes, myeloproliferative disorders, non-Hodgkin's lymphoma, Hodgkin's disease, chronic lymphocytic leukemia, multiple myeloma, juvenile chronic myeloid leukemia.

[0040] In some embodiments, said subject suffers from a non-malignant disorders selected from aplastic anemia, paroxysmal nocturnal hemoglobinuria, Fanconi's anemia, Blackfan-Diamond anemia, sickle cell anemia, thalassemia major, severe combined immunodeficiency, Wiskott- Aldrich syndrome, inborn errors of metabolism.

[0041] In some embodiments, said subject suffers from a disease selected from non-Hodgkin's lymphoma, chronic lymphocytic leukemia, multiple myeloma, acute myeloid leukemia, myelodysplastic syndromes, sickle cell anemia and thalassemia major.

[0042] In some embodiments, said subject suffers from non-Hodgkin's lymphoma.

[0043] In some embodiments, said subject suffers from chronic lymphocytic leukemia.

[0044] In some embodiments, said subject suffers from multiple myeloma.

[0045] In some embodiments, said subject suffers from acute myeloid leukemia.

[0046] In some embodiments, said subject suffers from myelodysplastic syndromes.

[0047] In some embodiments, said subject suffers from sickle cell anemia.

[0048] In some embodiments, said subject suffers from thalassemia major.

[0049] In some embodiments, said subject has a planned conditioning regimen in the context of an allogeneic HSCT.

[0050] In some embodiments, said subject is a high-risk patient, e.g. one with mismatched and / or unrelated donor, higher degrees of HLA disparity, older age or previous episodes of severe GvHD.

[0051] In some embodiments, said subject is a low-risk patients, e.g. one with a matched sibling donor, lower HLA disparity and no prior history of severe GvHD. In some embodiments, said subject is primarily affected or at risk to be primarily affected in the gastrointestinal tract with symptoms like severe diarrhea or abdominal pain.

[0052] In some embodiments, said subject is responsive to steroid treatment.

[0053] In some embodiments, said subject is not responsive to steroid treatment.

[0054] In some embodiments, said subject is responsive to treatment with ruxolitinib.

[0055] In some embodiments, said subject is not responsive to treatment with ruxolitinib.

[0056] In some embodiments, said subject is responsive to treatment with upadacitinib.

[0057] In some embodiments, said subject is not responsive to treatment with upadacitinib.

[0058] In some embodiments, said subject is receiving standard prophylaxis, e.g. standard immunosuppressive agents like calcineurin inhibitors (e.g., cyclosporine, tacrolimus).

[0059] In some embodiments, said subject has a matched related donor (MRD), i.e. a transplant from a HLA-matched family member.

[0060] In some embodiments, said subject has a mismatched related donor (MMRD), i.e. a transplant from a HLA-mismatched family member.

[0061] In some embodiments, said subject has a matched unrelated donor (MUD), i.e. a transplant from a HLA-matched unrelated donor.

[0062] In some embodiments, said subject has a mismatched unrelated donor (MMUD), i.e. a transplant from HLA-mismatched unrelated donor.

[0063] In some embodiments, said subject has a haploidentical donor, o.e. a transplant from a partially matched family member.

[0064] In some embodiments, said subject receives a cord blood transplantation, i.e. using stem cells derived from umbilical cord blood.

[0065] In some embodiments, said subject is a pediatric patient.

[0066] In some embodiments, said subject is an adult patient.

[0067] In some embodiments, said subject is a geriatric patient.

[0068] In some embodiments, said subject is characterized by having high levels of inflammatory cytokines and immune cell activation markers.

[0069] In some embodiments, said subject is characterized by having low levels of inflammatory cytokines and immune cell activation markers.

[0070] In some embodiments, said subject is selected based on higher levels of plasma regenerating islet-derived 3 alpha (REG3a).

[0071] In some embodiments, the higher levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is increased by about 1.2-, about 1.3-, about 1.4-, about 1.5-, about 2-, about 3- , about 4-, about 5-, about 6-, about 7-, about 8-, about 9-, about 10-, about 15-, about 20-, about 25-, about 30-, about 35-, about 40-, about 45-, about 50-, about 60-, about 70-, about 80-, about 90-, about 100-fold, or more, compared to normal population and / or baseline level.

[0072] In some embodiments, the higher levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is increased by about 1.2-fold compared to normal population and / or baseline level. In some embodiments, the higher levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is increased by about 2-fold compared to normal population and / or baseline level.

[0073] In some embodiments, the higher levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is increased by about 3-fold compared to normal population and / or baseline level.

[0074] In some embodiments, the higher levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is increased by about 5-fold compared to normal population and / or baseline level.

[0075] In some embodiments, the higher levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is increased by about 10-fold compared to normal population and / or baseline level.

[0076] In some embodiments, the higher levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is increased by at least 1.2-, at least 1.3-, at least 1.4-, at least 1.5-, at least 2-, at least 3-, at least 4-, at least 5-, at least 6-, at least 7-, at least 8-, at least 9-, at least 10-, at least at least 15-, at least 20-, at least 25-, at least 30-, at least 35-, at least 40-, at least 45-, at least 50-, at least 60-, at least 70-, at least 80-, at least 90-, at least 100-fold, or more, compared to normal population and / or baseline level.

[0077] In some embodiments, the higher levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is increased by at least 1.2-fold compared to normal population and / or baseline level.

[0078] In some embodiments, the higher levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is increased by at least 2-fold compared to normal population and / or baseline level.

[0079] In some embodiments, the higher levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is increased by at least 3-fold compared to normal population and / or baseline level.

[0080] In some embodiments, the higher levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is increased by at least 5-fold compared to normal population and / or baseline level.

[0081] In some embodiments, the higher levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is increased by at least 10-fold compared to normal population and / or baseline level.

[0082] In some embodiments, levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is > 45 ng / mL, > 72 ng / mL, > 88 ng / mL, > 100 ng / mL, > 120 ng / mL, > 150 ng / mL, or > 180 ng / mL.

[0083] In some embodiments, levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is > 45 ng / mL.

[0084] In some embodiments, levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is > 72 ng / mL. In some embodiments, levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is > 120 ng / mL.

[0085] In some embodiments, levels of plasma regenerating islet-derived 3 alpha (REG3a) in the subject is > 150 ng / mL.

[0086] In some embodiments, said subject is selected based on a biomarker selected from soluble IL-2 receptor alpha (slL-2Ra), TNF receptor 1 (TNFR1), interleukin-6 (IL-6), elafin, soluble suppression of tumorigenicity 2 (sST2), hepatocyte growth factor (HGF), chemokine ligand 9 (CXCL9), chemokine ligand 10 (CXCL10), annexin Al, lactate dehydrogenase (LDH), ferritin, albumin, C-reactive protein (CRP), beta-2 microglobulin, B-cell activating factor (BAFF), soluble CD13 and CD163.

[0087] In some embodiments, said subject is a female patient of reproductive potential.

[0088] In some embodiments, said subject is a female patient of reproductive potential, comprising the step of verifying the pregnancy status of the patient prior to initiating treatment.

[0089] In some embodiments, said subject is a female patient who is pregnant.

[0090] In some embodiments, said subject is a male patient of reproductive potential.

[0091] In one embodiment, the present invention refers to a compound of Formula (I), in particular the compound of Formula (II) or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof (Compound), for use in a patient who experiences one or more treatment-related, adverse reactions while receiving a clinically-recommended dose of the Compound comprising: ceasing administration of the Compound to the patient if the patient experiences at least one adverse reaction at a Grade 3 level (according to Common Terminology Criteria for Adverse Events (CTCAE)) or higher; and resuming administration of the Compound at a clinically recommended dose for the patient if the at least one adverse reaction did not rise to a Grade 4 level and the patient recovered to a Grade 1 level or less after treatment was ceased.

[0092] In one embodiment, the present invention refers to the Compound for use in a patient, comprising the steps of: administering the Compound to the patient at a clinically recommended dose for the patient; monitoring the patient for adverse reactions; ceasing administration of the Compound to the patient if the patient experiences at least one adverse reaction at a Grade 3 level; and resuming administration of the Compound to the patient when the patient's at least one adverse reaction has recovered to Grade 1 or less.

[0093] In some embodiments, the patient's at least one adverse reaction has recovered to Grade 0 after cessation of Compound administration, and administration is resumed.

[0094] In some embodiments, the patient's at least one adverse reaction is Grade 4, wherein administration of the Compound to the patient is permanently discontinued.

[0095] In some embodiments, the patient is monitored for adverse reactions over a period of time; in some embodiments, the monitoring occurs daily; in some embodiments, at least daily; in some embodiments, the monitoring occurs weekly; in some embodiments, at least monthly; in some embodiments, the monitoring occurs monthly; in some embodiments, at least monthly. In some embodiments, a clinically recommended dose for the patient that is in the range of 60 mg to 240 mg a day prior to and after the patient experiences one or more treatment- related, adverse reactions.

[0096] In some embodiments, a clinically recommended dose for the patient that is in the range of 60 mg to 240 mg mg taken once daily with food, either before and / or after the one or more adverse reactions.

[0097] In some embodiments, a clinically recommended dose for the patient that is reduced after the patient recovers to a Grade 1 level or less and administration of the Compound is resumed, for example, as compared with the clinically recommended dose administered to the patient before the at least one adverse reaction arose.

[0098] In some embodiments, the patient is being treated to prevent GvHD; in some embodiments, the patient is being treated for GvHD wherein the patient has failed at least one prior line of systemic therapy for the GvHD; in some embodiments, the patient is being treated for GvHD wherein the patient has failed at least two prior lines of systemic therapy for the GvHD.

[0099] In some embodiments, the patient is being treated to prevent aGvHD; in some embodiments, the patient is being treated for aGvHD wherein the patient has failed at least one prior line of systemic therapy for the aGvHD; in some embodiments, the patient is being treated for aGvHD wherein the patient has failed at least two prior lines of systemic therapy for the aGvHD.

[0100] In some embodiments, the patient is being treated for cGvHD; in some embodiments, the patient is being treated for cGvHD wherein the patient has failed at least one prior line of systemic therapy for the cGvHD; in some embodiments, the patient is being treated for cGvHD wherein the patient has failed at least two prior lines of systemic therapy for the cGvHD.

[0101] In some embodiments, the present invention provides for use of the Common Terminology Criteria for Adverse Events (CTCAE), for assessing or grading the severity of a patient's adverse reactions; in some embodiments, the CTCAE version is 5.0.

[0102] In some embodiments, the patient experiences at least one adverse reaction that is an infection, asthenia, nausea, diarrhea, dyspnea, cough, edema, hemorrhage, abdominal pain, musculoskeletal pain, headache, phosphate decrease, gamma glutamyl transferase increase, lymphocytes decrease, or hypertension.

[0103] In some embodiments, the patient experiences at least one adverse reaction that is a viral infection, a bacterial infection, or an infection of unspecified pathogen.

[0104] In some embodiments, the patient experiences at least one adverse reaction that is an infection of unspecified pathogen which is acute sinusitis, device related infection, ear infection, folliculitis, gastroenteritis, gastrointestinal infection, hordeolum, infectious colitis, lung infection, skin infection, tooth infection, urinary tract infection, wound infection, upper respiratory tract infection, pneumonia, conjunctivitis, sinusitis, respiratory tract infection, bronchitis, sepsis, or septic shock.

[0105] In some embodiments, the patient experiences at least one adverse reaction that is asthenia, edema, or pyrexia. In some embodiments, the patient experiences at least one adverse reaction that is a gastrointestinal disorder; in some embodiments, the gastrointestinal disorder is nausea, diarrhea, abdominal pain, or dysphagia.

[0106] In some embodiments, the patient experiences at least one adverse reaction that is a respiratory, thoracic or mediastinal disorder; in some embodiments, dyspnea, cough, or nasal congestion.

[0107] In some embodiments, the patient experiences at least one adverse reaction that is a vascular disorder; in some embodiments, hemorrhage or hypertension.

[0108] In some embodiments, the patient experiences at least one adverse reaction that is a musculoskeletal or connective tissue disorder; in some embodiments, the musculoskeletal or connective tissue disorder is musculoskeletal pain, muscle spasm, or arthralgia.

[0109] In some embodiments, the patient experiences at least one adverse reaction that is a nervous system disorder; in some embodiments, headache or migraine headache.

[0110] In some embodiments, the patient experiences at least one adverse reaction that is a metabolic disorder; in some embodiments, the metabolic disorder is decreased appetite.

[0111] In some embodiments, the patient experiences a skin or subcutaneous disorder; in some embodiments, the skin or subcutaneous disorder is a rash or pruritus.

[0112] In other embodiments, the patient experiences one or more treatment-related, adverse reactions while receiving a clinically- recommended dose of the Compound, comprising: ceasing administration of the Compound to the patient if the patient experiences at least one adverse reaction at a Grade 3 level or higher; and resuming administration of the Compound at a clinically recommended dose for the patient if the at least one adverse reaction did not rise to a Grade 4 level and the patient recovered to a Grade 1 level or less after treatment was ceased.

[0113] In some embodiments, the patient experiences at least one adverse reaction at a Grade 3 level and resuming administration of the Compound to the patient when the patient's at least one adverse reaction has recovered to Grade 1 or less.

[0114] In some embodiments, the patient experiences at least one adverse reaction at a Grade 4 level and permanently ceasing administration of the Compound to the patient.

[0115] In some embodiments, the subject has had allogeneic hematopoietic stem cell transplantation that is a matched-HSCT. In some embodiments, the allogeneic hematopoietic stem cell transplantation is a haploidentical-HSCT. In some embodiments, the allogeneic hematopoietic stem cell transplantation is a mismatched-HSCT.

[0116] In some embodiments, treatment with Compound is continued based on the patient's tolerability and intended to prevent GvHD. The number of cycles and duration of the treatment is patient dependent. In some embodiments, the Compound is administered to the patient up to 180 days post HSCT.

[0117] In some embodiments, treatment with Compound is continued based on the patient's tolerability until active aGvHD symptoms resolve or progress. The number of cycles and duration of the treatment is patient dependent. In some embodiments, the Compound is administered to the patient in one or more 28-day cycles. In some embodiments, the number of cycles ranges from 3 to 15. In some embodiments, the number of cycles ranges from 3 to 14, from 3 to 13, from 3 to 12, from 3 to 11, from 3 to 10, from 3 to 9, from 3 to 8, from 3 to 7, from 3 to 6, from 3 to 5, or from 3 to 4. In some embodiments, the number of cycles ranges from 5 to 11. In some embodiments, the number of cycles ranges from 6 to 12. In some embodiments, the number of cycles ranges from 5 to 10, from 5 to 9, or from 5 to 8. In some embodiments, the number of cycles ranges from 5 to 7. In some embodiments, the number of cycles ranges from 5 to 6. In some embodiments, the number of cycles is 5. In some embodiments, the number of cycles is 6. In some embodiments, the number of cycles is 7. In some embodiments, the number of cycles is 3, 4,

[0118] 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15.

[0119] In some embodiments, the number of cycles ranges from 3 cycles to loss of response. In some embodiments, the number of cycles ranges from 4 cycles to loss of response. In some embodiments, the number of cycles ranges from 5 cycles to loss of response. In some embodiments, the number of cycles ranges from 6 cycles to loss of response. In some embodiments, the number of cycles ranges from 7 cycles to loss of response. In some embodiments, the number of cycles ranges from 8 cycles to loss of response. In some embodiments, the number of cycles is greater than 3, 4, 5, 10, 15, 20, 25, or 30, or until a desired response is achieved.

[0120] In some embodiments, treatment with Compound is continued based on the patient's tolerability until active chronic GVHD symptoms resolve or progress. The number of cycles and duration of the treatment is patient dependent. In some embodiments, the Compound is administered to the patient up to 180 days.

[0121] In some embodiments, the subject experiences an improvement as defined by the Lee Symptom Scale (LSS). In some embodiments, the subject experiences at least a 7-point reduction in the LSS score. In some embodiments, the subject experiences at least a 10-point reduction in the LSS score. In some embodiments, the improvement is maintained over at least two consecutive evaluations. In some embodiments the LSS score is evaluated at baseline and on day 1 of each cycle starting at cycle 2 day 1.

[0122] In some embodiments, the subject has cGvHD and has failed one to three prior lines of systemic therapy for the cGvHD. In some embodiments, the subject has cGvHD and has failed at least two prior lines of systemic therapy for the cGvHD. In some embodiments, the subject has cGvHD and has failed two to five prior lines of systemic therapy for the cGvHD. In some embodiments, the subject has failed at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten prior lines of systemic therapy for the cGvHD.

[0123] In some embodiments, the subject experienced a complete response to last treatment for the GvHD prior to taking the Compound. In some embodiments, the subject experienced a partial response to last treatment for the GvHD prior to taking the Compound. In some embodiments, the subject experienced stable disease during the last treatment for the GvHD prior to taking the Compound. In some embodiments, the prior lines of systemic therapy for the cGvHD have been discontinued.

[0124] In some embodiments, the prior lines of systemic therapy are selected from the group consisting of prednisone, tacrolimus, ECP, sirolimus, ibruitinib, ruxolitinib, upadacitinib, MMF, rituximab, MTX, cyclosporine, imatinib, ixazomib, belumosudil and ofatumumab.

[0125] In some embodiments, the cGvHD is steroid-refractory (SR) cGvHD. In some embodiments, the subject is refractory to the last line of treatment prior to Compound treatment.

[0126] In some embodiments, the subject is receiving concomitant corticosteroid therapy. In some embodiments, the concomitant corticosteroid therapy is selected from the group consisting of prednisone, prednisolone, methylprednisolone, and budesonide. In some embodiments, the concomitant corticosteroid therapy is prednisone. In some embodiments, the dose of the concomitant corticosteroid therapy is reduced after at least 1 cycle of the Compound treatment. In some embodiments, the dose of the concomitant corticosteroid therapy is reduced by at least about 10%, by at least about 20%, by at least about 30%, by at least about 40%, by at least about 50%, by at least about 60%, or by at least about 70% after at least 1 cycle of the Compound treatment. In some embodiments, the dose of the concomitant corticosteroid therapy is reduced by from about 10% to about 70%, from about 15% to about 65%, from about 20% to about 60%, from about 30% to about 60%, from about 35% to about 60%, from about 40% to about 60%, or from about 45% to about 55% after at least 1 cycle of the Compound treatment. In some embodiments, the concomitant corticosteroid therapy is discontinued after at least 1 cycle of the Compound treatment.

[0127] In some embodiments, the subject is receiving concomitant calcineurin inhibitor therapy.

[0128] In some embodiments, the subject has involvement of at least 4 organs. In some embodiments, the subject has involvement of at least 3 organs. In some embodiments, the subject has involvement of at least 2 organs.

[0129] In some embodiments, the subject has involvement of the gastrointestinal tract.

[0130] In some embodiments, a compound of Formula (I), in particular the compound of Formula (II) or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof (Compound), is administered together with a second medication to alleviate the symptoms of GvHD. In some embodiments, the Compound and the second medication are administered simultaneously. In some embodiments, the Compound and the second medication are administered sequentially. The second medication can be, without limitation, a small molecule, an antibody or antigen binding fragment thereof, or radiation.

[0131] In some embodiments, administration of the Compound and the second medication provides an improvement in one or more symptoms of GvHD (e.g. as measured by the GvHD score) in the subject, compared to the one or more symptoms prior to administration of the Compound and the second medication.

[0132] In some embodiments, administration of the Compound and the second medication reduces or stops progression of one or more symptoms of the disease or disorder provided herein. In some embodiments, administration of the Compound and the second medication reduces or stops progression of one or more symptoms of GvHD (e.g. as measured by the GvHD score) in the subject, compared to the one or more symptoms prior to administration of the Compound and the second medication.

[0133] In some embodiments, administration of the Compound and the second medication provides a reduction in the GvHD score of the subject compared to the GvHD score of the subject prior to administration of the Compound and the second medication. For example, in some embodiments administration of the Compound and the second medication provides a reduction in the GvHD score of the subject from 4 to 3, from 4 to 2, from 4 to 1, from 4 to 0, from 3 to 2, from 3 to 1, from 3 to 0, from 2 to 1, from 2 to 0, or from 1 to 0.

[0134] In some embodiments, administration of the Compound and the second medication provides an improvement in one or more of weight loss of the subject, posture of the subject, activity of the subject, hair texture of the subject, and skin integrity of the subject, as measured by the GvHD score, compared to the weight loss of the subject, posture of the subject, activity of the subject, hair texture of the subject, and skin integrity of the subject prior to administration of the Compound and the second medication.

[0135] In some embodiments, administration of the Compound and the second medication provides an improvement in weight loss of the subject as measured by the GvHD score, compared to the weight loss of the subject as measured by the GvHD score, prior to administration of the Compound and the second medication. For example, in a subject exhibiting >15% weight loss prior to administration of the Compound and the second medication (Grade 4 as measured by the GvHD score), then administration of the Compound and the second medication may reduce the weight loss of the subject to between 10% and 15% (Grade 3 as measured by the GvHD score), between 5% and 10% (Grade 2 as measured by the GvHD score), between 0% and 5% (Grade 1 as measured by the GvHD score), or no weight losss (Grade 0 as measured by the GvHD score).

[0136] In some embodiments, administration of the Compound and the second medication provides an improvement in volume of diarrhea of the subject as measured by the GvHD score, compared to the volume of diarrhea of the subject as measured by the GvHD score, prior to administration of the Compound and the second medication.

[0137] In some embodiments, administration of the Compound and the second medication provides an improvement in total bilirubin level of the subject as measured by the GvHD score, compared to the total bilirubin level of the subject as measured by the GvHD score, prior to administration of the Compound and the second medication.

[0138] In some embodiments, administration of the Compound and the second medication provides an improvement in the posture of the subject as measured by the GvHD score, compared to the posture of the subject as measured by the GvHD score, prior to administration of the Compound and the second medication. For example, in a subject exhibiting severe hunched posture prior to administration of the Compound and the second medication (Grade 3 as measured by the GvHD score), then administration of the Compound and the second medication may improve the posture of the subject to a moderate hunched posture (Grade 2 as measured by the GvHD score), a mild hunched posture (Grade 1 as measured by the GvHD score), or a normal posture (Grade 0 as measured by the GvHD score).

[0139] In some embodiments, administration of the Compound and the second medication provides an improvement in the activity of the subject as measured by the GvHD score, compared to the activity of the subject as measured by the GvHD score, prior to administration of the Compound and the second medication. For example, in a subject exhibiting immobility prior to administration of the Compound and the second medication (Grade 3 as measured by the GvHD score), then administration of the Compound and the second medication may improve the activity of the subject to a slowed gait with refusal to move when touched (Grade 2 as measured by the GvHD score), a slowed gait (Grade 1 as measured by the GvHD score), or a normal activity (Grade 0 as measured by the GvHD score).

[0140] In some embodiments, the present invention relates to a compound of Formula (I) Formula (I), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in prevention or treatment of graft-versus-host disease, comprising administering to a subject in need thereof an effective amount of the compound according to Formula (I) or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof, optionally in combination with one or more additional therapeutically active compounds.

[0141] In some embodiments, the present invention relates to compound or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in prevention or treatment of graft-versus-host disease in a subject in need thereof.

[0142] In some embodiments, the present invention relates to compound or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in prevention or treatment of graft-versus-host disease in a subject in need thereof, comprising administering to a patient in need thereof an effective amount of this compound, optionally in combination with one or more additional therapeutically active compounds.

[0143] In some embodiments, the present invention relates to compound or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for the manufacture of a medicament for use in prevention or treatment of graft -versus-host disease in a subject in need thereof.

[0144] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject in a daily dose of 34 pmol to 1370 pmol, preferably of about 68 pmol to about 411 pmol, more preferably of about 137 pmol to about 411 pmol, and even more preferably of about 256 pmol to about 411 pmol.

[0145] In some embodiments the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject in a daily dose of 68 pmol to 411 pmol, preferably of about 137 pmol to about 411 pmol, and more preferably of about 256 pmol to about 411 pmol.

[0146] In some embodiments the compound of Formula (II) is administered to said subject in a daily dose of about 68 pmol, about 102 pmol, about 137 pmol, about 256 pmol or about 411 pmol, preferably in a daily dose of about 256 pmol or about 411 pmol.

[0147] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject in a daily dose from about 20 mg to about 800 mg, or from about 80 mg to about 320 mg, from about 60 mg to about 240 mg, from about 150 mg to about 240 mg, or from about 60 mg to about 160 mg. Exemplary unit doses include about 20 mg, about 40 mg, about 60 mg, about 80 mg, about 100 mg, about 120 mg, about 150 mg, about 160 mg, about 200 mg, about 240 mg, about 280 mg, about 300 mg, about 320 mg, about 400 mg, about 480 mg, about 500 mg, or about 600 mg.

[0148] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof result in a plasma trough level which is above 20 ng / mL at a steady state. For example, administration the compound of Formula (II) may result in a plasma trough level above 20 ng / mL, above 40 ng / mL, above 60 ng / mL, above 80 ng / mL, above 100 ng / mL, above 120 ng / mL, above 140 ng / mL, above 160 ng / mL, above 180 ng / mL, above 200 ng / mL, above 240 ng / mL, above 280 ng / mL, above 320 ng / mL, above 400 ng / mL, or above 500 ng / mL.

[0149] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof result in a plasma trough level, which is above 20 ng / mL at a steady state. For example, administration of the compound of Formula (II) may result in a plasma trough level range from about 20 ng / mL to 1000 ng / mL, from about 40 ng / mL to 500 ng / mL, from about 60 ng / mLto 320 ng / mL, from about 60 ng / mL to 200 ng / mL, from about 60 ng / mL to 200 ng / mL, from about 160 ng / mL to 320 ng / mL, from about 60 ng / mL to 90 ng / mL, from about 70 ng / mL to 85 ng / mL, from about 140 ng / mL to 200 ng / mL, or from about 150 ng / mL to 190 ng / mL.

[0150] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof, the steady state is achieved between about 3 to 14 days, about 5 to 10 days, or about 8 days after periodic or regular administration to a optionally once-a-day dosing of the drug, therapeutic combination or pharmaceutical dosage form.

[0151] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof, the plasma concentration for the drug, therapeutic combination or pharmaceutical dosage form is: (a) the trough level or trough concentration (Ctrough), or the lowest concentration reached by the drug, therapeutic combination or pharmaceutical dosage form before a second or next dose is administered, or (b) determined from blood samples taken between about 2 hours to 24 hours, or 4 to 12 hours, or 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more hours, after the last dose or administration of the drug, therapeutic combination or pharmaceutical dosage form.

[0152] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject with at least one additional therapeutic target.

[0153] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject with at least one immunosupressive agent.

[0154] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject with at least one immunosupressive or modulating agent which is selected from the group consisting of prednisone, prednisolone, methylprednisolone, budesonide, beclomethasone, dexamethasone, tacrolimus, sirolimus (rapamycin), cyclosporine, rifaximin, acalabrutinib, anlotinib, baricitinib, bortezomib, carfilzomib, entospletinib, fostamatinib, glasdegib, ibruitinib, imatinib, itacitinib, ixazomib (MLN9708), jaktinib, nilotinib, nintedanib, pacritinib, pimicotinib, ruxolitinib, upadacitinib, sonidegib, vismodegib, other JAK inhibitors (e.g. SHR0302), axatilimab, alemtuzumab, basiliximab, belimumab, brentuximab vedotin, daclizumab, efalizumab, gavilimomab (ABX- CBL), ibritumomab tiuxetan, infliximab, inolimomab, itolizumab, milatuzumab, muromonab- CD3, natalizumab, neihulizumab, obinutuzumab, ofatumumab, rituximab, siplizumab, tildrakizumab, tocilizumab, ustekinumab, vedolizumab, visilizumab, etanercept, alefacept, abatacept, IL-2, teduglutide, lenalidomide, pomalidomide, leflunomide, thalidomide, panobinostat (LBH589), alvelestat (MPH966), vorinostat, mycophenolate mofetil (MMF), methotrexate (MTX), decitabine, clofarabine, melphalan, thiotepa, pentostatin, palifermin, filgrastim, prochymal (remestemcel-L), cyclophosphamide, defibrotide, cannabidiol, VM-001 (from ViGenCell), RGI-2001, ASC930, TQ05105 (or other JAK / ROCK inhibitors), GDC-8264, ALPN-101, voriconazole, itraconazole, opebacan, belumosudil (KD025), efmarodocokin alfa, efavaleukin alfa (AMG 592), efprezimod alfa, romidepsin, busulfan, fludarabine phosphate, aldesleukin, thymoglobulin, methoxsalen, extracorporeal photopheresis (ECP), mesenchymal stromal cells (e.g. CYP-001, OTI-010 or MC0518), allogeneic faecal microbiota (e.g. MaaT013), fecal microbiota trasnplantation, BET inhibitor (e.g. PLX51107), C5a inhibitor (e.g. ALXN1007), sitagliptin, atorvastatin, clobetasol, maraviroc, ribaxamase (SYN-004), sargramostim, alpha 1- antitrypsin, mitoxantrone or combinations thereof.

[0155] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject with at least one steroid, e.g. prednisone, prednisolone, methylprednisolone, budesonide, beclomethasone or dexamethasone.

[0156] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject with at least ruxolitinib.

[0157] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject with at least upadacitinib.

[0158] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject with at least belumosudil.

[0159] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject for a period of at least 4 weeks, preferably at least 14 weeks, more preferably at least 26 weeks post HSCT, and most preferably longer.

[0160] In some emdodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered orally.

[0161] In some embodiments, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof may be administered for a prolonged period, for example, for at least about 1 week, at least about 2 weeks, at least about 3 weeks, at least about 4 weeks, at least about 5 weeks, at least about 6 weeks, at least about 7 weeks, at least about 8 weeks, at least about 9 weeks, at least about 10 weeks, at least about 11 weeks, at least about 12 weeks, at least about 14 weeks or at least about 26 weeks post HSCT. For example, the compound of Formula (II) may be administered for about 2 weeks, at least about 3 weeks, at least about 4 weeks, at least about 5 weeks, at least about 6 weeks, at least about 7 weeks at least about 8 weeks, at least about 9 weeks, at least about 10 weeks, at least about 11 weeks, at least about 14 weeks, or at least about 26 weeks post HSCT. In some embodiments, the compound of Formula (II) is administered for at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 12 months post HSCT, or longer.

[0162] In another aspect, the present invention relates to a method of prevention or treatment of graft-versus-host disease, comprising administering to a subject in need thereof an effective among of the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof.

[0163] In another aspect, the present invention relates to a method of prevention or treatment of graft-versus-host disease, comprising administering to a subject in need thereof an effective among of the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof.

[0164] In another aspect, the present invention relates to a method of prevention or treatment of graft-versus-host disease, comprising administering to a subject in need thereof an effective among of the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof, wherein said subject suffers from a disease selected from the group comprising.

[0165] In some embodiments of said method, said compound of Formula (I), in particular the compound of Formula (II), is capable of restoring intestinal barrier function, inducing physiologic regeneration of the crypt cells.

[0166] In some embodiments of said method, said subject is human.

[0167] In some embodiments of said method, said subject suffers from a disease selected from chronic GvHD (cGvHD), acute GvHD (aGvHD), gastrointestinal GvHD, steroid-refractory GvHD, steroid-resistant GvHD or transfusion-associated graft-versus-host disease.

[0168] In some embodiments of said method, said subject suffers from gastrointestinal GvHD.

[0169] In some embodiments of said method, said subject suffers from steroid-refractory GvHD or steroid-resistant GvHD.

[0170] In some embodiments of said method, said subject is at risk of developing aGvHD.

[0171] In some embodiments of said method, said subject suffers from a disease selected from malignant disorders (such as acute myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, myelodysplastic syndromes, myeloproliferative disorders, non-Hodgkin's lymphoma, Hodgkin's disease, chronic lymphocytic leukemia, multiple myeloma, juvenile chronic myeloid leukemia), non-malignant disorders (such as aplastic anemia, paroxysmal nocturnal hemoglobinuria, Fanconi's anemia, Blackfan-Diamond anemia, sickle cell anemia, thalassemia major, severe combined immunodeficiency, Wiskott-Aldrich syndrome, inborn errors of metabolism) or a planned conditioning regimen in the context of an allogeneic HSCT. In some embodiments of said method, said subject suffers from a malignant disorders selected from acute myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, myelodysplastic syndromes, myeloproliferative disorders, non-Hodgkin's lymphoma, Hodgkin's disease, chronic lymphocytic leukemia, multiple myeloma, juvenile chronic myeloid leukemia.

[0172] In some embodiments of said method, said subject suffers from a non-malignant disorders selected from aplastic anemia, paroxysmal nocturnal hemoglobinuria, Fanconi's anemia, Blackfan-Diamond anemia, sickle cell anemia, thalassemia major, severe combined immunodeficiency, Wiskott-Aldrich syndrome, inborn errors of metabolism.

[0173] In some embodiments of said method, said subject suffers from a disease selected from nonHodgkin's lymphoma, chronic lymphocytic leukemia, multiple myeloma, acute myeloid leukemia, myelodysplastic syndromes, sickle cell anemia and thalassemia major.

[0174] In some embodiments of said method, said subject suffers from non-Hodgkin's lymphoma.

[0175] In some embodiments of said method, said subject suffers from chronic lymphocytic leukemia.

[0176] In some embodiments of said method, said subject suffers from multiple myeloma.

[0177] In some embodiments of said method, said subject suffers from acute myeloid leukemia.

[0178] In some embodiments of said method, said subject suffers from myelodysplastic syndromes.

[0179] In some embodiments of said method, said subject suffers from sickle cell anemia.

[0180] In some embodiments of said method, said subject suffers from thalassemia major.

[0181] In some embodiments of said method, said subject has a planned conditioning regimen in the context of an allogeneic HSCT.

[0182] In some embodiments of said method, said subject is a high-risk patient, e.g. one with mismatched and / or unrelated donor, higher degrees of HLA disparity, older age or previous episodes of severe GvHD.

[0183] In some embodiments of said method, said subject is a low-risk patients, e.g. one with a matched sibling donor, lower HLA disparity and no prior history of severe GvHD.

[0184] In some embodiments of said method, said subject is primarily affected or at risk to be primarily affected in the gastrointestinal tract with symptoms like severe diarrhea or abdominal pain.

[0185] In some embodiments of said method, said subject is responsive to steroid treatment.

[0186] In some embodiments of said method, said subject is not responsive to steroid treatment. In some embodiments of said method, said subject is responsive to treatment with ruxolitinib. In some embodiments of said method, said subject is not responsive to treatment with ruxolitinib. In some embodiments of said method, said subject is responsive to treatment with upadacitinib.

[0187] In some embodiments of said method, said subject is not responsive to treatment with upadacitinib.

[0188] In some embodiments of said method, said subject is receiving standard prophylaxis, e.g. standard immunosuppressive agents like calcineurin inhibitors (e.g., cyclosporine, tacrolimus).

[0189] In some embodiments of said method, said subject has a matched related donor (MRD), i.e. a transplant from a HLA-matched family member.

[0190] In some embodiments of said method, said subject has a mismatched related donor (MMRD), i.e. a transplant from a HLA-mismatched family member.

[0191] In some embodiments of said method, said subject has a matched unrelated donor (MUD), i.e. a transplant from a HLA-matched unrelated donor.

[0192] In some embodiments of said method, said subject has a mismatched unrelated donor (MMUD), i.e. a transplant from HLA-mismatched unrelated donor.

[0193] In some embodiments of said method, said subject has a haploidentical donor, o.e. a transplant from a partially matched family member.

[0194] In some embodiments of said method, said subject receives a cord blood transplantation, i.e. using stem cells derived from umbilical cord blood.

[0195] In some embodiments of said method, said subject is a pediatric patient.

[0196] In some embodiments of said method, said subject is an adult patient.

[0197] In some embodiments of said method, said subject is a geriatric patient.

[0198] In some embodiments of said method, said subject is characterized by having high levels of inflammatory cytokines and immune cell activation markers.

[0199] In some embodiments of said method, said subject is characterized by having low levels of inflammatory cytokines and immune cell activation markers.

[0200] In some embodiments of said method, said subject is selected based on higher levels of plasma regenerating islet-derived 3 alpha (REG3a).

[0201] In some embodiments of said method, said subject is selected based on a biomarker selected from soluble IL-2 receptor alpha (slL-2Ra), TNF receptor 1 (TNFR1), interleukin-6 (IL-6), elafin, soluble suppression of tumorigenicity 2 (sST2), hepatocyte growth factor (HGF), chemokine ligand 9 (CXCL9), chemokine ligand 10 (CXCL10), annexin Al, lactate dehydrogenase (LDH), ferritin, albumin, C-reactive protein (CRP), beta-2 microglobulin, B-cell activating factor (BAFF), soluble CD13 and CD163.

[0202] In some embodiments of said method, said subject is a female patient of reproductive potential.

[0203] In some embodiments of said method, said subject is a female patient of reproductive potential, comprising the step of verifying the pregnancy status of the patient prior to initiating treatment.

[0204] In some embodiments of said method, said subject is a female patient who is pregnant.

[0205] In some embodiments of said method, said subject is a male patient of reproductive potential. In one embodiment of said method, the patient experiences one or more treatment-related, adverse reactions while receiving a clinically-recommended dose of the Compound comprising: ceasing administration of the Compound to the patient if the patient experiences at least one adverse reaction at a Grade 3 level (according to Common Terminology Criteria for Adverse Events (CTCAE)) or higher; and resuming administration of the Compound at a clinically recommended dose for the patient if the at least one adverse reaction did not rise to a Grade 4 level and the patient recovered to a Grade 1 level or less after treatment was ceased.

[0206] In one embodiment said method comprises the steps of: administering the Compound to the patient at a clinically recommended dose for the patient; monitoring the patient for adverse reactions; ceasing administration of the Compound to the patient if the patient experiences at least one adverse reaction at a Grade 3 level; and resuming administration of the Compound to the patient when the patient's at least one adverse reaction has recovered to Grade 1 or less.

[0207] In some embodiments of said method, the patient's at least one adverse reaction has recovered to Grade 0 after cessation of Compound administration, and administration is resumed.

[0208] In some embodiments of said method, the patient's at least one adverse reaction is Grade 4, wherein administration of the Compound to the patient is permanently discontinued.

[0209] In some embodiments of said method, the patient is monitored for adverse reactions over a period of time; in some embodiments, the monitoring occurs daily; in some embodiments, at least daily; in some embodiments, the monitoring occurs weekly; in some embodiments, at least monthly; in some embodiments, the monitoring occurs monthly; in some embodiments, at least monthly.

[0210] In some embodiments of said method, a clinically recommended dose for the patient that is in the range of 60 mg to 240 mg a day prior to and after the patient experiences one or more treatment-related, adverse reactions.

[0211] In some embodiments of said method, a clinically recommended dose for the patient that is in the range of 60 mg to 240 mg mg taken once daily with food, either before and / or after the one or more adverse reactions.

[0212] In some embodiments of said method, a clinically recommended dose for the patient that is reduced after the patient recovers to a Grade 1 level or less and administration of the Compound is resumed, for example, as compared with the clinically recommended dose administered to the patient before the at least one adverse reaction arose.

[0213] In some embodiments of said method, the patient is being treated to prevent GvHD; in some embodiments, the patient is being treated for GvHD wherein the patient has failed at least one prior line of systemic therapy for the GvHD; in some embodiments, the patient is being treated for GvHD wherein the patient has failed at least two prior lines of systemic therapy for the GvHD. In some embodiments of said method, the patient is being treated to prevent aGvHD; in some embodiments, the patient is being treated for aGvHD wherein the patient has failed at least one prior line of systemic therapy for the aGvHD; in some embodiments, the patient is being treated for aGvHD wherein the patient has failed at least two prior lines of systemic therapy for the aGvHD.

[0214] In some embodiments of said method, the patient is being treated for cGvHD; in some embodiments, the patient is being treated for cGvHD wherein the patient has failed at least one prior line of systemic therapy for the cGvHD; in some embodiments, the patient is being treated for cGvHD wherein the patient has failed at least two prior lines of systemic therapy for the cGvHD.

[0215] In some embodiments of said method, the Common Terminology Criteria for Adverse Events (CTCAE) is used for assessing or grading the severity of a patient's adverse reactions; in some embodiments, the CTCAE version is 5.0.

[0216] In some embodiments of said method, the patient experiences at least one adverse reaction that is an infection, asthenia, nausea, diarrhea, dyspnea, cough, edema, hemorrhage, abdominal pain, musculoskeletal pain, headache, phosphate decrease, gamma glutamyl transferase increase, lymphocytes decrease, or hypertension.

[0217] In some embodiments of said method, the patient experiences at least one adverse reaction that is a viral infection, a bacterial infection, or an infection of unspecified pathogen.

[0218] In some embodiments of said method, the patient experiences at least one adverse reaction that is an infection of unspecified pathogen which is acute sinusitis, device related infection, ear infection, folliculitis, gastroenteritis, gastrointestinal infection, hordeolum, infectious colitis, lung infection, skin infection, tooth infection, urinary tract infection, wound infection, upper respiratory tract infection, pneumonia, conjunctivitis, sinusitis, respiratory tract infection, bronchitis, sepsis, or septic shock.

[0219] In some embodiments of said method, the patient experiences at least one adverse reaction that is asthenia, edema, or pyrexia.

[0220] In some embodiments of said method, the patient experiences at least one adverse reaction that is a gastrointestinal disorder; in some embodiments, the gastrointestinal disorder is nausea, diarrhea, abdominal pain, or dysphagia.

[0221] In some embodiments of said method, the patient experiences at least one adverse reaction that is a respiratory, thoracic or mediastinal disorder; in some embodiments, dyspnea, cough, or nasal congestion.

[0222] In some embodiments of said method, the patient experiences at least one adverse reaction that is a vascular disorder; in some embodiments, hemorrhage or hypertension.

[0223] In some embodiments of said method, the patient experiences at least one adverse reaction that is a musculoskeletal or connective tissue disorder; in some embodiments, the musculoskeletal or connective tissue disorder is musculoskeletal pain, muscle spasm, or arthralgia.

[0224] In some embodiments of said method, the patient experiences at least one adverse reaction that is a nervous system disorder; in some embodiments, headache or migraine headache. In some embodiments of said method, the patient experiences at least one adverse reaction that is a metabolic disorder; in some embodiments, the metabolic disorder is decreased appetite.

[0225] In some embodiments of said method, the patient experiences a skin or subcutaneous disorder; in some embodiments, the skin or subcutaneous disorder is a rash or pruritus.

[0226] In other embodiments of said method, the present disclosure provides a method of treating a patient who experiences one or more treatment-related, adverse reactions while receiving a clinically- recommended dose of the Compound, comprising: ceasing administration of the Compound to the patient if the patient experiences at least one adverse reaction at a Grade 3 level or higher; and resuming administration of the Compound at a clinically recommended dose for the patient if the at least one adverse reaction did not rise to a Grade 4 level and the patient recovered to a Grade 1 level or less after treatment was ceased.

[0227] In some embodiments of said method, the patient experiences at least one adverse reaction at a Grade 3 level and resuming administration of the Compound to the patient when the patient's at least one adverse reaction has recovered to Grade 1 or less.

[0228] In some embodiments of said method, the patient experiences at least one adverse reaction at a Grade 4 level and permanently ceasing administration of the Compound to the patient.

[0229] In some embodiments of said method, the subject has had allogeneic hematopoietic stem cell transplantation that is a matched-HSCT. In some embodiments, the allogeneic hematopoietic stem cell transplantation is a haploidentical-HSCT. In some embodiments, the allogeneic hematopoietic stem cell transplantation is a mismatched-HSCT.

[0230] In some embodiments, treatment with Compound is continued based on the patient's tolerability and intended to prevent GvHD. The number of cycles and duration of the treatment is patient dependent. In some embodiments, the Compound is administered to the patient up to 180 days post HSCT.

[0231] In some embodiments of said method, treatment with Compound is continued based on the patient's tolerability until active aGvHD symptoms resolve or progress. The number of cycles and duration of the treatment is patient dependent. In some embodiments, the Compound is administered to the patient in one or more 28-day cycles.

[0232] In some embodiments of said method, the number of cycles ranges from 3 to 15. In some embodiments, the number of cycles ranges from 3 to 14, from 3 to 13, from 3 to 12, from 3 to 11, from 3 to 10, from 3 to 9, from 3 to 8, from 3 to 7, from 3 to 6, from 3 to 5, or from 3 to 4. In some embodiments, the number of cycles ranges from 5 to 11. In some embodiments, the number of cycles ranges from 6 to 12. In some embodiments, the number of cycles ranges from 5 to 10, from 5 to 9, or from 5 to 8. In some embodiments, the number of cycles ranges from 5 to 7. In some embodiments, the number of cycles ranges from 5 to 6. In some embodiments, the number of cycles is 5. In some embodiments, the number of cycles is 6. In some embodiments, the number of cycles is 7. In some embodiments, the number of cycles is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15.

[0233] In some embodiments of said method, the number of cycles ranges from 3 cycles to loss of response. In some embodiments, the number of cycles ranges from 4 cycles to loss of response. In some embodiments, the number of cycles ranges from 5 cycles to loss of response. In some embodiments, the number of cycles ranges from 6 cycles to loss of response. In some embodiments, the number of cycles ranges from 7 cycles to loss of response. In some embodiments, the number of cycles ranges from 8 cycles to loss of response. In some embodiments, the number of cycles is greater than 3, 4, 5, 10, 15, 20, 25, or 30, or until a desired response is achieved.

[0234] In some embodiments of said method, treatment with Compound is continued based on the patient's tolerability until active chronic GVHD symptoms resolve or progress. The number of cycles and duration of the treatment is patient dependent. In some embodiments, the Compound is administered to the patient up to 180 days.

[0235] In some embodiments of said method, the subject experiences an improvement as defined by the Lee Symptom Scale (LSS). In some embodiments, the subject experiences at least a 7- point reduction in the LSS score. In some embodiments, the subject experiences at least a 10- point reduction in the LSS score. In some embodiments, the improvement is maintained over at least two consecutive evaluations. In some embodiments the LSS score is evaluated at baseline and on day 1 of each cycle starting at cycle 2 day 1.

[0236] In some embodiments of said method, the subject has cGvHD and has failed one to three prior lines of systemic therapy for the cGvHD. In some embodiments, the subject has cGvHD and has failed at least two prior lines of systemic therapy for the cGvHD. In some embodiments, the subject has cGvHD and has failed two to five prior lines of systemic therapy forthe cGvHD. In some embodiments, the subject has failed at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten prior lines of systemic therapy for the cGvHD.

[0237] In some embodiments of said method, the subject experienced a complete response to last treatment for the GvHD prior to taking the Compound. In some embodiments, the subject experienced a partial response to last treatment for the GvHD prior to taking the Compound. In some embodiments, the subject experienced stable disease during the last treatment for the GvHD prior to taking the Compound.

[0238] In some embodiments of said method, the prior lines of systemic therapy for the cGvHD have been discontinued.

[0239] In some embodiments of said method, the prior lines of systemic therapy are selected from the group consisting of prednisone, tacrolimus, ECP, sirolimus, ibruitinib, ruxolitinib, upadacitinib, MMF, rituximab, MTX, cyclosporine, imatinib, ixazomib, belumosudil and ofatumumab.

[0240] In some embodiments of said method, the cGvHD is steroid-refractory (SR) cGvHD. In some embodiments, the subject is refractory to the last line of treatment prior to Compound treatment.

[0241] In some embodiments of said method, the subject is receiving concomitant corticosteroid therapy. In some embodiments, the concomitant corticosteroid therapy is selected from the group consisting of prednisone, prednisolone, methylprednisolone, and budesonide. In some embodiments, the concomitant corticosteroid therapy is prednisone. In some embodiments, the dose of the concomitant corticosteroid therapy is reduced after at least 1 cycle of the Compound treatment. In some embodiments, the dose of the concomitant corticosteroid therapy is reduced by at least about 10%, by at least about 20%, by at least about 30%, by at least about 40%, by at least about 50%, by at least about 60%, or by at least about 70% after at least 1 cycle of the Compound treatment. In some embodiments, the dose of the concomitant corticosteroid therapy is reduced by from about 10% to about 70%, from about 15% to about 65%, from about 20% to about 60%, from about 30% to about 60%, from about 35% to about 60%, from about 40% to about 60%, or from about 45% to about 55% after at least 1 cycle of the Compound treatment. In some embodiments, the concomitant corticosteroid therapy is discontinued after at least 1 cycle of the Compound treatment.

[0242] In some embodiments of said method, the subject is receiving concomitant calcineurin inhibitor therapy.

[0243] In some embodiments of said method, the subject has involvement of at least 4 organs. In some embodiments, the subject has involvement of at least 3 organs. In some embodiments, the subject has involvement of at least 2 organs.

[0244] In some embodiments of said method, the subject has involvement of the gastrointestinal tract.

[0245] In some embodiments of said method, a compound of Formula (I), in particularthe compound of Formula (II) or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof (Compound), is administered together with a second medication to alleviate the symptoms of GvHD. In some embodiments, the Compound and the second medication are administered simultaneously. In some embodiments, the Compound and the second medication are administered sequentially. The second medication can be, without limitation, a small molecule, an antibody or antigen binding fragment thereof, or radiation.

[0246] In some embodiments of said method, administration of the Compound and the second medication provides an improvement in one or more symptoms of GvHD (e.g. as measured by the GvHD score) in the subject, compared to the one or more symptoms prior to administration of the Compound and the second medication.

[0247] In some embodiments of said method, administration of the Compound and the second medication reduces or stops progression of one or more symptoms of the disease or disorder provided herein.

[0248] In some embodiments of said method, administration of the Compound and the second medication reduces or stops progression of one or more symptoms of GvHD (e.g. as measured by the GvHD score) in the subject, compared to the one or more symptoms prior to administration of the Compound and the second medication.

[0249] In some embodiments of said method, administration of the Compound and the second medication provides a reduction in the GvHD score of the subject compared to the GvHD score of the subject prior to administration of the Compound and the second medication. For example, in some embodiments administration of the Compound and the second medication provides a reduction in the GvHD score of the subject from 4 to 3, from 4 to 2, from 4 to 1, from 4 to 0, from 3 to 2, from 3 to 1, from 3 to 0, from 2 to 1, from 2 to 0, or from 1 to 0.

[0250] In some embodiments of said method, administration of the Compound and the second medication provides an improvement in one or more of weight loss of the subject, posture of the subject, activity of the subject, hair texture of the subject, and skin integrity of the subject, as measured by the GvHD score, compared to the weight loss of the subject, posture of the subject, activity of the subject, hair texture of the subject, and skin integrity of the subject prior to administration of the Compound and the second medication.

[0251] In some embodiments of said method, administration of the Compound and the second medication provides an improvement in weight loss of the subject as measured by the GvHD score, compared to the weight loss of the subject as measured by the GvHD score, prior to administration of the Compound and the second medication. For example, in a subject exhibiting >15% weight loss prior to administration of the Compound and the second medication (Grade 4 as measured by the GvHD score), then administration of the Compound and the second medication may reduce the weight loss of the subject to between 10% and 15% (Grade 3 as measured by the GvHD score), between 5% and 10% (Grade 2 as measured by the GvHD score), between 0% and 5% (Grade 1 as measured by the GvHD score), or no weight losss (Grade 0 as measured by the GvHD score).

[0252] In some embodiments of said method, administration of the Compound and the second medication provides an improvement in volume of diarrhea of the subject as measured by the GvHD score, compared to the volume of diarrhea of the subject as measured by the GvHD score, prior to administration of the Compound and the second medication.

[0253] In some embodiments of said method, administration of the Compound and the second medication provides an improvement in total bilirubin level of the subject as measured by the GvHD score, compared to the total bilirubin level of the subject as measured by the GvHD score, prior to administration of the Compound and the second medication.

[0254] In some embodiments of said method, administration of the Compound and the second medication provides an improvement in the posture of the subject as measured by the GvHD score, compared to the posture of the subject as measured by the GvHD score, prior to administration of the Compound and the second medication. For example, in a subject exhibiting severe hunched posture prior to administration of the Compound and the second medication (Grade 3 as measured by the GvHD score), then administration of the Compound and the second medication may improve the posture of the subject to a moderate hunched posture (Grade 2 as measured by the GvHD score), a mild hunched posture (Grade 1 as measured by the GvHD score), or a normal posture (Grade 0 as measured by the GvHD score). In some embodiments, administration of the Compound and the second medication provides an improvement in the activity of the subject as measured by the GvHD score, compared to the activity of the subject as measured by the GvHD score, prior to administration of the Compound and the second medication. For example, in a subject exhibiting immobility prior to administration of the Compound and the second medication (Grade 3 as measured by the GvHD score), then administration of the Compound and the second medication may improve the activity of the subject to a slowed gait with refusal to move when touched (Grade 2 as measured by the GvHD score), a slowed gait (Grade 1 as measured by the GvHD score), or a normal activity (Grade 0 as measured by the GvHD score).

[0255] In some embodiments, the present invention relates to a method of prevention or treatment of graft-versus-host disease, comprising administering to a subject in need thereof an effective among of the compound of Formula (I) Formula (I), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof, optionally in combination with one or more additional therapeutically active compounds.

[0256] In some embodiments, the present invention relates to a method of prevention or treatment of graft-versus-host disease, comprising administering to a subject in need thereof an effective among of compound or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof, optionally in combination with one or more additional therapeutically active compounds.

[0257] In some embodiments of said methods, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject in a daily dose of 34 pmol to 1370 pmol, preferably of about 68 pmol to about 411 pmol, more preferably of about 137 pmol to about 411 pmol, and even more preferably of about 256 pmol to about 411 pmol.

[0258] In some embodiments of said methods, the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject in a daily dose of 68 pmol to 411 pmol, preferably of about 137 pmol to about 411 pmol, and more preferably of about 256 pmol to about 411 pmol.

[0259] In some embodiments the compound of Formula (II) is administered to said subject in a daily dose of about 68 pmol, about 102 pmol, about 137 pmol, about 256 pmol or about 411 pmol, preferably in a daily dose of about 256 pmol or about 411 pmol.

[0260] In some embodiments of said methods, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject in a daily dose from about 20 mg to about 800 mg, or from about 80 mg to about 320 mg, from about 60 mg to about 240 mg, from about 150 mg to about 240 mg, or from about 60 mg to about 160 mg. Exemplary unit doses include about 20 mg, about 40 mg, about 60 mg, about 80 mg, about 100 mg, about 120 mg, about 150 mg, about 160 mg, about 200 mg, about 240 mg, about 280 mg, about 300 mg, about 320 mg, about 400 mg, about 480 mg, about 500 mg, or about 600 mg.

[0261] In some embodiments of said methods, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof result in a plasma trough level which is above 20 ng / mL at a steady state. For example, administration the compound of Formula (II) may result in a plasma trough level above 20 ng / mL, above 40 ng / mL, above 60 ng / mL, above 80 ng / mL, above 100 ng / mL, above 120 ng / mL, above 140 ng / mL, above 160 ng / mL, above 180 ng / mL, above 200 ng / mL, above 240 ng / mL, above 280 ng / mL, above 320 ng / mL, above 400 ng / mL, or above 500 ng / mL.

[0262] In some embodiments of said methods, treating the subject with the compound of Formula (II) resulted in a plasma trough level, which is above 20 ng / mL at a steady state. For example, administration of the compound of Formula (II) may result in a plasma trough level range from about 20 ng / mL to 1000 ng / mL, from about 40 ng / mL to 500 ng / mL, from about 60 ng / mL to 320 ng / mL, from about 60 ng / mL to 200 ng / mL, from about 60 ng / mL to 200 ng / mL, from about 160 ng / mL to 320 ng / mL, from about 60 ng / mL to 90 ng / mL, from about 70 ng / mL to 85 ng / mL, from about 140 ng / mL to 200 ng / mL, or from about 150 ng / mL to 190 ng / mL.

[0263] In some embodiments of said methods, the steady state is achieved between about 3 to 14 days, about 5 to 10 days, or about 8 days after periodic or regular administration to a optionally once-a-day dosing of the drug, therapeutic combination or pharmaceutical dosage form.

[0264] In some embodiments of said methods, the plasma concentration for the drug, therapeutic combination or pharmaceutical dosage form is: (a) the trough level or trough concentration (Ctrough), or the lowest concentration reached by the drug, therapeutic combination or pharmaceutical dosage form before a second or next dose is administered, or (b) determined from blood samples taken between about 2 hours to 24 hours, or 4 to 12 hours, or 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more hours, after the last dose or administration of the drug, therapeutic combination or pharmaceutical dosage form.

[0265] In some embodiments of said methods, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject for a period of at least 12 weeks, preferably at least 26 weeks, more preferably at least 52 weeks.

[0266] In some emdodiments of said methods, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered orally.

[0267] In some embodiments of said methods, the compound of Formula (I), in particular the compound of Formula (II), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof may be administered for a prolonged period, for example, for at least about 1 week, at least about 2 weeks, at least about 3 weeks, at least about 4 weeks, at least about 5 weeks, at least about 6 weeks, at least about 7 weeks, at least about 8 weeks, at least about 9 weeks, at least about 10 weeks, at least about 11 weeks, at least about 12 weeks, at least about 14 weeks or at least about 26 weeks post HSCT. For example, the compound of Formula (II) may be administered for about 2 weeks, at least about 3 weeks, at least about 4 weeks, at least about 5 weeks, at least about 6 weeks, at least about 7 weeks at least about 8 weeks, at least about 9 weeks, at least about 10 weeks, at least about 11 weeks, at least about 14 weeks, or at least about 26 weeks post HSCT. In some embodiments, the compound of Formula (II) is administered for at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 12 months post HSCT, or longer.

[0268] The present invention further relates relates to use of the compound of Formula (I) Formula (I), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof, for manufacturing a medicament for prevention or treatment of graft-versus-host disease in a subject in need thereof, optionally in combination with one or more additional therapeutically active compounds.

[0269] The present invention further relates relates to use of the compound of Formula (II) Formula (II) , or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof, for manufacturing a medicament for prevention or treatment of graft-versus-host disease, optionally in combination with one or more additional therapeutically active compounds.

[0270] In embodiments disclosed herein, including in the uses and methods of treatment provided herein, the clinically recommended dose for the patient may be in the range of 60 mg a day to 240 mg a day; in some embodiments, the clinically recommended dose for the patient is 150 mg a day; in some embodiments, the clinically recommended dose for the patient is 240 mg a day; in some embodiments, the clinically recommended dose forthe patient is a reduced dose after the patient recovers to a Grade 1 level or less and administration of the Compound is resumed, as compared with the clinically recommended dose administered before the patient experienced at least one adverse reaction. It will be recognized that features specified in each embodiment may be combined with other specified features to provide further embodiments of the present disclosure:

[0271] 1. A compound of Formula (I) Formula (I), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft -versus- host disease in a subject in need thereof, wherein:

[0272] R1is selected from the group consisting of hydrogen, carboxyl, cyano, fluorine, chlorine, methyl, isopropyl, t-butyl, trifluoromethyl, trifluoromethoxy, cyclopropylmethoxy, 1,1- difluoro-2-methylpropyl, l,l-difluoro-2,2-dimethylpropyl, 1-methyl-l-cyclobutyl, methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl, 1-hydroxy-l-methylethyl, azetidine-l-carbonyl, 3-methyloxetan-3-yl, 4,5-dihydrooxazol-2-yl and cyclopropylcarbonyl;

[0273] R2is hydrogen or methyl;

[0274] A1is =N- or =CH-;

[0275] X is benzene, pyridine or cyclohexane;

[0276] J is any ring selected from the group

[0277] Y is selected from the group consisting of phenyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, piperidinyl, azetidinyl, tetra hydropyranyl, morpholinyl and tetrahydropyridinyl, optionally substituted with 1, 2 or 3 groups selected from Yl, or alternatively Y is amino, optionally substituted with 1 or 2 groups selected from Yl;

[0278] Y1is selected from the group consisting of hydroxy, cyano, fluorine, amino, methyl, ethyl, isopropyl, methoxy, azetidinyl, pyrrolidinyl, morpholinyl, , wherein amino is optionally substituted with 1 or 2 groups selected from Y2and methyl, ethyl, isopropyl, methoxy, azetidinyl, pyrrolidinyl, morpholinyl, are optionally substituted with 1, 2 or 3 groups selected from Y2;

[0279] Y2is selected from the group consisting of hydroxyl, fluorine, amino, methyl, methoxy, azetidinyl, pyrrolidinyl, morpholinyl, wherein amino is optionally substituted with 1 or 2 groups selected from Y3and methyl, methoxy, azetidinyl, pyrrolidinyl, morpholinyl, is optionally substituted with 1, 2 or 3 groups selected from Y3,

[0280] Y3is hydroxyl, fluorine, cyano, methyl or methoxy, wherein methyl or methoxy are optionally substituted with fluorine. A compound of Formula (I) or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof according to embodiment 1, wherein the compound is selected from the following group:

[0281] A compound of Formula (I) or an isotopic variant, a pharmacologically acceptable salt thereof ora solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof according to embodiment 1 or 2, wherein the compound is Formula (II). The compound of Formula (II), which is for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof. A compound of Formula (I) or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft -versus-host disease in a subject in need thereof according to any of embodiments 1 to 4, wherein said subject is primarily affected or at risk to be primarily affected in the gastrointestinal tract with symptoms like severe diarrhea or abdominal pain. A compound of Formula (I) or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft -versus-host disease in a subject in need thereof according to any of embodiments 1 to 5, wherein said subject suffers from a disease selected from malignant disorders (such as acute myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, myelodysplastic syndromes, myeloproliferative disorders, nonHodgkin's lymphoma, Hodgkin's disease, chronic lymphocytic leukemia, multiple myeloma, juvenile chronic myeloid leukemia) or non-malignant disorders (such as aplastic anemia, paroxysmal nocturnal hemoglobinuria, Fanconi's anemia, Blackfan- Diamond anemia, sickle cell anemia, thalassemia major, severe combined immunodeficiency, Wiskott-Aldrich syndrome, inborn errors of metabolism). A compound of Formula (I) or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof according to any of embodiments 1 to 6, wherein said subject has a planned conditioning regimen in the context of an allogeneic HSCT. A compound of Formula (I) or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft -versus-host disease in a subject in need thereof according to any of embodiments 1 to 7, wherein the subject is selected based on higher levels of plasma regenerating islet-derived 3 alpha (REG3a). A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft -versus-host disease in a subject in need thereof according to any of embodiments 1 to 8, wherein the graft -versus-host disease is selected from chronic GvHD (cGvHD), acute GvHD (aGvHD), gastrointestinal GvHD, steroid-refractory GvHD, steroid- resistant GvHD or transfusion-associated graft -versus-host disease. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft -versus-host disease in a subject in need thereof according to any of embodiments 1 to 9, wherein said compound or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject in a daily dose of 34 pmol to 1370 pmol, preferably of about 68 pmol to about 411 pmol, more preferably of about 137 pmol to about 411 pmol, and even more preferably of about 256 pmol to about 411 pmol. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft -versus-host disease in a subject in need thereof according to any of embodiments I to 10, wherein said compound or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject with at least one additional therapeutic treatment. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft -versus-host disease in a subject in need thereof according to any of embodiment 11, wherein said at least one additional therapeutic treatment is an immunosupressive treatment. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof according to embodiment 12, wherein said immunosupressive treatment is selected from the group consisting of prednisone, prednisolone, methylprednisolone, budesonide, beclomethasone, dexamethasone, tacrolimus, sirolimus (rapamycin), cyclosporine, rifaximin, acalabrutinib, anlotinib, baricitinib, bortezomib, carfilzomib, entospletinib, fostamatinib, glasdegib, ibruitinib, imatinib, itacitinib, ixazomib (MLN9708), jaktinib, nilotinib, nintedanib, pacritinib, pimicotinib, ruxolitinib, upadacitinib, sonidegib, vismodegib, other JAK inhibitors (e.g. SHR0302), axatilimab, alemtuzumab, basiliximab, belimumab, brentuximab vedotin, daclizumab, efalizumab, gavilimomab (ABX-CBL), ibritumomab tiuxetan, infliximab, inolimomab, itolizumab, milatuzumab, muromonab- CD3, natalizumab, neihulizumab, obinutuzumab, ofatumumab, rituximab, siplizumab, tildrakizumab, tocilizumab, ustekinumab, vedolizumab, visilizumab, etanercept, alefacept, abatacept, IL-2, teduglutide, lenalidomide, pomalidomide, leflunomide, thalidomide, panobinostat (LBH589), alvelestat (MPH966), vorinostat, mycophenolate mofetil (MMF), methotrexate (MTX), decitabine, clofarabine, melphalan, thiotepa, pentostatin, palifermin, filgrastim, prochymal (remestemcel-L), cyclophosphamide, defibrotide, cannabidiol, VM-001 (from ViGenCell), RGI-2001, ASC930, TQ05105 (or other JAK / ROCK inhibitors), GDC-8264, ALPN-101, voriconazole, itraconazole, opebacan, belumosudil (KD025), efmarodocokin alfa, efavaleukin alfa (AMG 592), efprezimod alfa, romidepsin, busulfan, fludarabine phosphate, aldesleukin, thymoglobulin, methoxsalen, extracorporeal photopheresis (ECP), mesenchymal stromal cells (e.g. CYP-001, OTI-OIO or MC0518), allogeneic faecal microbiota (e.g. MaaT013), fecal microbiota trasnplantation, BET inhibitor (e.g. PLX51107), C5a inhibitor (e.g. ALXN1007), sitagliptin, atorvastatin, clobetasol, maraviroc, ribaxamase (SYN-004), sargramostim, alpha 1- antitrypsin, mitoxantrone or combinations thereof. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof according to embodiment 13, wherein said agent is selected from prednisone, prednisolone, methylprednisolone, budesonide, beclomethasone, dexamethasone, ruxolitinib, upadacitinib, belumosudil or combinations thereof. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof according to any of embodiments I to 14, wherein said compound or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject for a period of at least 4 weeks, preferably at least 14 weeks, more preferably at least 26 weeks post HSCT. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of treatment of graft -versus-host disease in a subject in need thereof according to any of embodiments 1 to 15, wherein said treatment selectively modulates SIRT6 activity. 1. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of treatment of graft -versus-host disease in a subject in need thereof according to any of embodiments 1 to 16, wherein said treatment causes selective SIRT6 increase.

[0282] DEFINITIONS

[0283] An "isotopic variant" refers within the scope of the invention to deuterated variations of compounds of Formula (I) or (II), or a pharmaceutically acceptable salt or solvate thereof. Usually, each molecule in the proportion of the population is substituted with a deuterium atom at the same position. The proportion of molecules of the population that comprise a deuterium atom at a specific atomic position can be represented by an isotopic enrichment factor sp / s, where £p / s = (Rp / Rs - 1) x 1000, where Rp is the abundance of deuterium at that position within the population of molecules, and Rs is the natural abundance of deuterium at the position. The proportion of molecules of the population that comprise a deuterium atom at a specific atomic position can alternatively be represented by the molar percent of the population that is substituted with deuterium at that atomic position (i.e., % deuterium incorporation). Non-limiting examples of an isotopic enrichment factor include at least 835 (12.5% deuterium incorporation), at least 1670 (25% deuterium incorporation), at least 3500 (52.5% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation). Non-limiting examples of an abundance of deuterium in a sample of a compound herein include a natural abundance, and abundance that is at least 3340 times of the natural abundance of deuterium, which is 0.015% (i.e., at least 50.1% incorporation of deuterium), at least 3500 times of the natural abundance of deuterium (52.5% deuterium incorporation), at least 4500 times of the natural abundance of deuterium (67.5% deuterium incorporation), at least 5000 (75% deuterium), at least 5500 times of the natural abundance of deuterium (82.5% deuterium incorporation), at least 6000 times of the natural abundance of deuterium (90% deuterium incorporation), at least 6333.3 times of the natural abundance of deuterium (95% deuterium incorporation), at least 6466.7 times of the natural abundance of deuterium (97% deuterium incorporation), at least 6600 times of the natural abundance of deuterium (99% deuterium incorporation), at least 6633.3 times of the natural abundance of deuterium (99.5% deuterium incorporation), or any other abundance. When a particular position in a compound of the invention (e.g., a compound represented by Formula (I) or (II), or a pharmaceutically acceptable salt and / or solvate thereof) is designated specifically by name or structure as "H" or "hydrogen", the position is understood to have hydrogen at its natural abundance isotopic composition. The percentage of deuterium incorporation can be obtained by quantitative analysis using, for example, mass spectroscopy (peak area) or by quantifying the remaining residual 1H-NMR signals of the specific deuteration site compared to signals from internal standards or other, nondeuterated 1H signals in the compound.

[0284] The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic acids, including inorganic acids and organic acids. Thus, the compound of the present disclosure contains a basic group and can be used according to the disclosure, for example, as hydrochloride, aspartate, glutamate, L-tatrate, malonate, fumarate, citrate, malate, maleate, lactate, gluconate, benzoate, succinate, acetate, phosphate, sulfate, napsylate, besylate, tosylate or mesylate salt. The respective salts can be obtained by customary methods which are known to the person skilled in the art like, for example, by contacting these with an organic or inorganic acid in a solvent or dispersant, or by anion exchange with other salts. The present disclosure also includes all salts of the compounds of the present disclosure which, owing to low physiological compatibility, are not directly suitable for use in pharmaceuticals but which can be used, for example, as intermediates for chemical reactions or for the preparation of pharmaceutically acceptable salts.

[0285] "Pharmaceutically acceptable" means suitable for use in a human subject.

[0286] The term "solvate" refers to a crystalline form of a molecule that further comprises molecules of a solvent or solvents incorporated into the crystalline latice structure. Thus, the compounds of the present disclosure may be present in the form of solvates, such as those which include as solvate water, or pharmaceutically acceptable solvates, such as alcohols, in particular ethanol. A stoichiometric or non-stoichiometric amount of solvent is bound by non- covalent intermolecular forces. When the solvent is water, the "solvate" is a "hydrate." It is understood, that a "pharmaceutically acceptable salts" can in addition optionally contain a "solvate".

[0287] "A week" preferably refers to a period of time of or about 5, about 6 or about 7 days. It may be about 5-8 days.

[0288] "A month" preferably refers to a period of time of or about 28, about 29, about 30 or about 31 days. It may be about 26-33 days.

[0289] The term "treating" or "treatment" means an alleviation of symptoms associated with a disease, disorder or condition, or halt of further progression or worsening of those symptoms. Depending on the disease and condition of the subject, the term "treatment" as used herein may include one or more of curative and palliative treatment. Treatment can also include administering a pharmaceutical formulation of the present invention in combination with other therapies.

[0290] The term "prevention" or "preventing" refers to the administration or application of a compound, composition, or treatment regimen in a manner that inhibits the onset or reduces the risk of occurrence of a specific disease or pathological condition in a subject, e.g. GvHD. This includes any intervention that precludes the initiation, progression, or recurrence of the disease, thereby maintaining the health and well-being of the subject. Prevention encompasses both prophylactic measures administered prior to any signs or symptoms of the disease and strategies aimed at preventing disease relapse or recurrence in individuals who have previously been treated for the condition.

[0291] "Daily dose" preferably refers to the total dose of a compound according to Formula (I) or (II) or an isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, preferably orally administered to the subject or patient each day of administration. The daily dose can be reached through a single or several administrations per day, such as for example once a day, twice a day or three times a day. Preferably, it is reached or achieved by single administration per day, preferably consisting of one or more tablets or capsules, preferably tablets or capsules as described herein.

[0292] As used herein, the term "effective amount" includes a dosage sufficient to produce a desired result with respect to the indicated disorder, condition, or mental state. The desired result may comprise a subjective or objective improvement in the recipient of the dosage.

[0293] As used herein, the term "administering" includes activities associated with providing a patient an amount of a compound according to Formula (I) or (II) or an isotopic variant, a pharmaceutically acceptable salt or a solvate thereof. Administering includes providing unit dosages of compositions set forth herein to a patient in need thereof. Administering includes providing effective amounts of compounds, e.g. the compound of Formula (II) or an isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, for a specified period of time, e.g. for about 6, 9, 12, 15 or more months, or about 1, 2, 3, 4, 5 or more years.

[0294] As used herein, the term "patient" or "subject" refers to refers to a living mammalian organism. In one embodiment the patient is a human subject.

[0295] As used herein, the term "administered as adjunct therapy" or "administered with at least one additional therapeutic target" includes sequential or simultaneous administration of two or more structurally different compounds. For example, two or more structurally different pharmaceutically active compounds can be co-administered by administering a pharmaceutical composition adapted for oral administration that contains two or more structurally different active pharmaceutically active compounds. As another example, two or more structurally different compounds can be co-administered by administering one compound and then administering the other (second) compound. In some instances, the coadministered compounds are administered by the same route. In other instances, the coadministered compounds are administered via different routes. For example, one compound can be administered orally, and the other compound can be administered, e.g. sequentially or simultaneously, via intravenous or intraperitoneal injection.

[0296] The term "about" as used herein with respect to numbers, figures, ranges and / or amounts is preferably meant to mean "circa" and / or "approximately". The meaning of those terms is well known in the art and preferably includes a variance, deviation and / or variability of the respective number, figure, range and / or amount of plus / minus 15% and especially of plus / minus 10%.

[0297] As used herein, the term "graft -versus-host disease" or "GvHD" refers to a condition, including acute and chronic, resulting from cellular transplanted effects (graft) in host cells and tissues resulting from GVH. In other words, donor immune cells infused into the graft or donor immune cells that develop from the stem cells, can see the patient's (host) cells as foreign and turn against them with an immune response. As examples, patients who have received a blood or bone marrow transplant from another person are at risk of having an acute GvHD. Acute graft versus host disease (GvHD) is specifically a disorder caused by donor immune cells in patients who have received an allogeneic bone marrow or blood cell transplant. The most commonly affected tissues are the skin, intestine and liver. In severe cases, GvHD can cause blistering of the skin or excessive diarrhea and wear. In addition, inflammation caused by donor immune cells in the liver can cause a blockage that causes a disease. Other tissues such as the lung and thymus may also be affected. The diagnosis is usually confirmed by observing a small piece of skin, liver, stomach or intestine with a microscope to observe specific inflammatory characteristics. In severe cases, the liver does not work properly to eliminate waste products from the body. Acute GvHD usually begins during the first 3 months after transplantation. In some cases, it may persist, return or begin more than 3 months after the transplant.

[0298] As used herein, the term "gastrointestinal graft-versus-host disease" refers to the damage caused by the donor's immune cells to house the stomach and intestine tissue that can cause loss of appetite, nausea, vomiting or diarrhea as part of the GvHD, whether acute or chronic. In severe cases, gastrointestinal graft-versus-host disease can cause pain in the abdomen and bleeding in the stomach or intestines. As used herein, the term "allogeneic transplant" refers to infusions of donor blood or spinal cord stem cell transplants from a donor to a host patient. In other words, the patient receives stem cells from the bone marrow or blood from a donor of similar tissue.

[0299] As used herein, the term "allogeneic HSCT" refers to the transplantation of multipotent hematopoietic stem cells from one individual to another. This allogeneic HSCT is performed in patients with certain cancers of the blood or bone marrow, such as multiple myeloma or leukemia, congenital immunodeficiencies, and bone marrow failures or other hematologic disease. In these cases, the recipient's immune system is usually destroyed with radiation or chemotherapy before transplantation. Infection and graft versus host disease are an important complication of HSCT.

[0300] When the term "grade" is used to refer to the grade level of an adverse reaction, the term is intended to be defined for most adverse event categories in accordance with Common Terminology Criteria for Adverse Events (CTCAE), currently version 5.0 scale. The CTCAE displays Grades 1 through 5 with unique clinical descriptions of severity for each adverse reaction based on the general guidelines: Grade 1 - Mild (asymptomatic or mild symptoms; clinical or diagnostic observations only; intervention not indicated); Grade 2 - Moderate (minimal, local or noninvasive intervention indicated); Grade 3 - Severe (medically significant but not immediately life-threatening; hospitalization or prolongation of hospitalization indicated; disabling; limiting self-care); Grade 4 - Life-threatening (urgent intervention indicated); and Grade 5 - Death.

[0301] Lee Symptom Scale (LSS) summary score measures the effect on patients' functioning and well-being. The Lee Symptom Scale is a 30-item scale developed to measure the symptoms of cGvHD and is described in Biol. Blood Marrow Transplant. 2002;8:444.

[0302] In the context of GvHD, "HLA disparity" refers to the differences or mismatches between the human leukocyte antigens (HLA) of the donor and the recipient (Biol. Blood Marrow Transplant. 2002;8:387). The lower the HLA disparity, the better the chances of survival.

[0303] In the context of GvHD, "conditioning treatment" refers to the preparative regimen given to a patient before undergoing a HSCT. This treatment typically involves chemotherapy, radiation therapy, or a combination of both, with the primary objectives of (a) eradicating malignant cells (in cases where the patient has a malignancy, such as leukemia or lymphoma, the conditioning regimen aims to eliminate cancerous cells); (b) suppressing the immune system (conditioning helps to suppress the patient's immune system to prevent rejection of the transplanted donor stem cells); and (c) creating space in the bone marrow (it creates an environment conducive to the engraftment of the donor stem cells by depleting the recipient's existing bone marrow cells). A common conditioning regimen might include agents for chemotherapy (like busulfan, cyclophosphamide) or monoclonal antibody therapy and / or total body irradiation, tailored to the patient's specific medical condition and the type of transplant (allogeneic or autologous). The intensity of conditioning regimens can vary, with some being myeloablative (fully ablative) and others being reduced-intensity or non- myeloablative, depending on the patient's health status and the underlying disease being treated.

[0304] Plasma levels of regenerating islet-derived 3 alpha (REG3a) protein can be determined e.g as described in Blood 2013;122:4602.

[0305] In the context of GvHD, "primarily affected" refers to the main organs or tissues that exhibit the most significant signs and symptoms of the disease. GvHD commonly targets specific organs, and when these organs (e.g. skin, gastrointestinal tract, liver or mouth) are "primarily affected," it means they are the primary sites of inflammation, damage, and clinical manifestations due to the immune response triggered by the transplanted donor cells.

[0306] In the context of SIRT6 modulation within the SIRT family, "selectively modulating" refers to an selectivity index of SIRT6 towards one or more members of the SIRT family of about 5-, about 10-, about 20-, about 50-, about 100-fold, about 500-fold, about 1000-fold, or more. The selectivity index can be determined e.g. by measuring the respective SIRT modulation at the same concentration for the respective member and generation of the quotient as outlined in Example 5.

[0307] Compositions may further comprise one or more pharmaceutically acceptable additional ingredient(s) such as alum, stabilizers, antimicrobial agents, buffers, coloring agents, flavoring agents, adjuvants, and the like.

[0308] Compositions may be in the form of tablets or lozenges formulated in a conventional manner. For example, tablets and capsules for oral administration may contain conventional excipients including, but not limited to, binding agents, fillers, lubricants, disintegrants and weting agents. Binding agents include, but are not limited to, syrup, acacia, gelatine, sorbitol, tragacanth, mucilage of starch and polyvinylpyrrolidone. Fillers include, but are not limited to, lactose, sugar, microcrystalline cellulose, maize starch, calcium phosphate, and sorbitol. Lubricants include, but are not limited to, magnesium stearate, stearic acid, talc, polyethylene glycol, and silica. Disintegrants include, but are not limited to, potato starch and sodium starch glycollate. Weting agents include, but are not limited to, sodium lauryl sulfate). Tablets may be coated according to methods well known in the art. Compositions may also be liquid formulations including, but not limited to, aqueous or oily suspensions, solutions, emulsions, syrups, and elixirs. The compositions may also be formulated as a dry product for constitution with water or other suitable vehicle before use, such liquid preparations may contain additives including, but not limited to, suspending agents, emulsifying agents, nonaqueous vehicles and preservatives. Suspending agent include, but are not limited to, sorbitol syrup, methyl cellulose, glucose / sugar syrup, gelatine, hydroxyethylcellulose, carboxymethyl cellulose, aluminium stearate gel, and hydrogenated edible fats. Emulsifying agents include, but are not limited to, lecithin, sorbitan monooleate, and acacia. Nonaqueous vehicles include, but are not limited to, edible oils, almond oil, fractionated coconut oil, oily esters, propylene glycol, and ethyl alcohol. Preservatives include, but are not limited to, methyl or propyl p-hydroxybenzoate and sorbic acid.

[0309] Combination treatments:

[0310] As already indicated, in all of the therapeutic methods or indications disclosed above, the compound of the present invention may be administered alone. However, it may also be administered in combination with one or more additional therapeutically active agents, substances or compounds, either sequentially or concomitantly.

[0311] As described above, compounds of the present invention may be administered or applied in combination with one or more additional therapeutically active compounds, substances or treatments, and suitable additional compounds, substances or treatments may be selected, for example, from extracorporeal photopheresis (ECP); anti-thymocyte globulin; alpha 1-antitrypsin; calcineurin inhibitors (e.g. cyclosporine, tacrolimus) antimetabolites (e.g. methotrexate, mycophenolate mophetyl); anti-CD20 monoclonal antibodies (e.g. rituximab, ofatumumab, obinutuzumab);

[0312] CSF-1R blocking monoclonal antibodies (e.g. axatilimab);

[0313] CD52-binding monoclonal antibodies (e.g. alemtuzumab);

[0314] CD26-binding monoclonal antibodies (e.g. begelomab); anti-TNFa antibodies (e.g. infliximab); anti-CD30 antibody-drug conjugate (e.g. brentuximab vedotin); deacetylase inhibitor (e.g. panobinostat); mTOR inhibitors (e.g. sirolimus, everolimus, temsirolimus, ridaforolimus) cytokine IL-2 or IL-2 muteins (e.g. AMG 592); tyrosine kinase inhibitors (e.g. imatinib, nilotinib, ibrutinib, fostamatinib, entospletinib), and the sub-group JAK1 / 2 inhibitors (e.g. tofacitinib, oclacitinib, filgotinib, momelotinib, ilginatinib, BMS-911543, INCB52793, peficitinib, WP1066, ivarmacitinib, brepocitinib, ritlecitinib, abrocitinib, itacitinib, ruxolitinib, baricitinib, upadacitinib) or hedgehog inhibitors (e.g. vismodegib, sonidegib); proteasome inhibitors (e.g. ixazomib, bortezomib, carfilzomib); rho-kinase 2 inhibitors (rho-associated protein kinase 2 inhibitor or ROCK2 inhibitor, e.g. belumosudil, SR3677, GV101) or Rho-kinase 1 / 2 inhibitors; mitogen activated extracellular signal regulated kinases 1 and 2 (MEK1 and MEK2) inhibitors (e.g. trametinib);

[0315] PI3K inhibitors (e.g. idelalisib, GS-649443); fusion proteins inhibiting the CD28 signaling pathway (e.g. Abatacept), inhibits T cell activation and proliferation (e.g. alefacept); fusion proteins inhibiting TNFa (e.g. etanercept); hydroxychloroquine; cytoprotective adjuvants or cytoprotectants (e.g. amifostine, mesna, palifermin, dexrazoxane, / V-acetylcysteine, glutamine, ursodeoxycholic acid); or chemotherapeutics (e.g. pentostatin).

[0316] DESCRIPTION OF FIGURES

[0317] Figure 1A shows the single-cycle-kinetic analysis of SIRT6 binding to immobilized compound 2 (without NAD) according to Example 3.

[0318] Figure IB illustrates the SIRT6 binding to immobilized compound 2 from Example 3 in absence and presence of NAD.

[0319] Figure 2 illustrates that compound 1 is a potent and highly selective inhibitor of SIRT6. Each recombinant human sirtuin protein was incubated with the corresponding biotinylated peptide, NAD+and indicated compound 1 concentrations at room temperature for 1 h. Changes in peptide modification were measured by AlphaLISA assay. The % inhibition was calculated from individual values of enzyme activities. The data are shown as the mean ± standard deviation (n=4).

[0320] EXAMPLES

[0321] Example 1: Chemical synthesis of the compound according to Formula (II)

[0322] Step 1: Methyl trans-4-(methylcarbamoyl)cyclohexane-l-carboxylate (II) trans-4-(Methoxycarbonyl)cyclohexane carboxylic acid (25.3 g) was dissolved in CH2CI2 (400 mL). To this, NEts (56.6 mL), methylamine hydrochloride (18.4 g), 3 / 7-l,2,3-triazolo[4,5- b] pyridin-3-ol (0.93 g) and l-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (29.5 g) were added and the mixture was stirred at room temperature (rt) for 70 h. To the mixture, water and IN HCI was added, and the mixture was extracted with CH2CI2. The organic layer was dried over anhydrous MgSCU. The resultant solution was concentrated under reduced pressure and the precipitated solid was diluted with hexane. The obtained solid was collected by filtration to yield compound II as a white solid.TH NMR (CDCI3) 6 (ppm) 1.38- 1.58 (m, 4H), 1.92-1.98 (m, 2H), 2.01-2.09 (m, 3H), 2.27-2.34 (m, 1H), 2.81 (d, J = 4.9 Hz, 3H), 3.67 (s, 3H), 5.40-5.48 (m, 1H).

[0323] Step 2: Methyl trans-4-(methylcarbamothioyl)cyclohexane-l-carboxylate (lk)

[0324] Compound II (19 g) was dissolved in toluene (400 mL). To this, Lawesson's reagent (21.3 g) was added and the mixture was stirred at 90°C for 8 h. To the mixture, saturated aqueous sodium bicarbonate was added and the mixture was extracted with EtOAc. The organic layer was washed with brine and dried over anhydrous MgSCU. The residue obtained by concentration under reduced pressure was purified by NH silica gel column chromatography (CF Cb / EtOAc = 9:1 to 1:1 (v / v)) to obtain compound lk as a white solid.1H NMR (CDCh) 6 (ppm) 1.44-1.54 (m, 2H), 1.69-1.79 (m, 2H), 1.93-1.98 (m, 2H), 2.07-2.12 (m, 2H), 2.31-2.38 (m, 1H), 2.41-2.48 (m, 1H), 3.19 (d, J = 4.9 Hz, 3H), 3.67 (s, 3H), 7.26-7.31 (m, 1H).

[0325] Step 3: 2-((4-(Trifluoromethyl)pyridin-2-yl)oxy)acetohydrazide (lj)

[0326] Methyl 2-((4-(trifluoromethyl)pyridin-2-yl)oxy)acetate (111 g) was dissolved in EtOH (2.4 L). To this, hydrazine monohydrate (177 g) was added and the mixture was stirred at 90°C for 3 h. The resultant solution was concentrated under reduced pressure to obtain compound lj as a white solid.

[0327] Step 4: Methyl trans-4-((methylimino)(methylthio)methyl)cyclohexane-l-carboxylate (li)

[0328] Compound lk (1.5 g) was dissolved in acetone (400 mL). To this, K2CO3 (26.5 g) and methyl iodide (15.9 mL) were added and the mixture was stirred for 5 h under reflux with heating. Afterthe reaction temperature was allowed to reach rt, concentrated under reduced pressure and diluted CH2CI2. The precipitated solid was collected by filtration to obtain compound li as a white solid.TH NMR (CDCI3) 6 (ppm) 1.44-1.56 (m, 4H), 1.7 9-1.85 (m, 0.7H), 1.88-1.97 (m,1.3H), 2.04-2.11 (m, 2H), 2.20 (s, 1H), 2.29-2.36 (m, 1H), 2.42 (s, 2H), 2.47-2.53 (m, 0.7H), 2.80-2.87 (m, 0.3H), 3.17 (s, 2H), 3.26 (s, 1H), 3.67 (s, 1.3H), 3.67 (s, 0.7H).

[0329] Step 5: Methyl trans-4-(4-methyl-5-(((4-(trifluoromethyl)pyridin-2-yl)oxy)methyl)-4 / 7-l,2,4- triazol-3-yl)cyclohexane-l-carboxylate (lh)

[0330] Compound li (20 g) was dissolved in EtOH (200 mL). To this, compound lj (20.5 g) was added and the mixture was stirred at 90°C for 4 h. The residue obtained by concentration under reduced pressure was purified by silica gel column chromatography (MeOH / CFhCb = 0:1 to 1:9 (v / v)) to obtain compound lh as a white solid.1H NMR (CDCI3) 6 (ppm) 1.52-1.64 (m, 2H), 1.87-1.98 (m, 2H), 2.06-2.14 (m, 2H), 2.16-2.23 (m, 2H), 2.47 (dt, J = 12.2, 3.4 Hz, 1H), 2.75 (dt, J = 12.2, 3.4 Hz, 1H), 3.70 (s, 3H), 3.78 (s, 3H), 5.40 (s, 2H), 7.22-7.32 (m, 3H), 7.45 (t, J = 7.8 Hz, 1H).

[0331] Step 6: (trans-4-(4-methyl-5-(((4-(trifluoromethyl)pyridin-2-yl)oxy)methyl)-4 / 7-l,2,4-triazol-

[0332] 3-yl)cyclohexyl)methanol (lg)

[0333] Compound lh (19.7 g) was dissolved in THF (250 mL). To this, a LiAIH4-THF solution (2.5M, 20 mL) was added under ice-cooling and the mixture was stirred at rt for 25 minutes. To the mixture, water (1.9 mL), a 15% aqueous NaOH solution (1.9 mL) and water (5.7 mL) were sequentially added under ice-cooling and the mixture was diluted with EtOAc and then filtered with celite. The filtrate was concentrated under reduced pressure to obtain compound lg as a light yellow oily substance.TH NMR (CDCI3) 6 (ppm) 1.08-1.20 (m, 2H), 1.34 (d, J = 5.4 Hz, 1H), 1.55-1.72 (m, 1H), 1.76-1.89 (m, 2H), 1.94-2.11 (m, 4H), 2.62 (dt, J = 12.2, 3.4 Hz, 1H), 3.53 (t, J = 5.9 Hz, 2H), 3.66 (s, 3H), 5.29 (s, 2H), 7.22-7.29 (m, 3H), 7.42 (t, J = 7.8 Hz, 1H).

[0334] Step 7: trans-4-(4-Methyl-5-(((4-(trifluoromethyl)pyridin-2-yl)oxy)methyl)-4 / 7-l,2,4-triazol-3- yl)cyclohexane-l-carbaldehyde (If)

[0335] Compound lg (17.9 g) was dissolved in CH2CI2 (300 mL). To this, Dess-Martin periodinane (22.4 g) was added under ice-cooling and the mixture was stirred for 5 minutes under icecooling and at rt for40 minutes. The solution was ice-cooled. To this, a IN NaOH solution (230 mL) and a 10% aqueous sodium thiosulfate solution were added and stirred, and then the mixture was extracted with CH2CI2. The organic layer was dried over anhydrous Na2SO4. The residue obtained by concentration under reduced pressure was purified by silica gel column chromatography (MeOH / EtOAc = 0:1 to 1:9 (v / v)) to obtain compound If as a white solid.TH NMR (CDCh) 6 (ppm) 1.36-1.47 (m, 2H), 1.82-1.92 (m, 2H), 2.09-2.16 (m, 2H), 2.18-2.25 (m, 2H), 2.36-2.44 (m, 1H), 2.64 (dt, J = 12.2, 3.4 Hz, 1H), 3.67 (s, 3H), 5.30 (s, 2H), 7.22-7.31 (m, 3H), 7.43 (t, J = 7.8 Hz, 1H), 9.70 (s, 1H).

[0336] Step 8: 2-(trans-4-(4-Methyl-5-(((4-(trifluoromethyl)pyridin-2-yl)oxy)methyl)-4 / 7-l,2,4- triazol-3-yl)cyclohexyl)acetaldehyde (le)

[0337] (Methoxymethyl)triphenylphosphonium chloride (13.2 g) was suspended in THF (100 mL). To this, potassium tert-butoxide (4.31 g) was added under ice-cooling and the mixture was stirred for 20 minutes. The resultant solution was added dropwise using a cannula to a solution of compound If (9.44 g) in THF (150 mL). After 45 minutes, to a suspension of (methoxymethyl)triphenylphosphonium chloride (8.79 g) in THF (100 mL), potassium tert- butoxide (2.88 g) was added under ice-cooling and the mixture was stirred for 5 minutes. The obtained solution was added dropwise again to the reaction solution using a cannula. After 10 minutes, water was added to the reaction mixture and the mixture was concentrated under reduced pressure and extracted with EtOAc. The organic layer was washed with brine and then dried over anhydrous Na2SO4. After filtration, the residue obtained by concentration under reduced pressure was purified by silica gel column chromatography (EtOAc / hexane / MeOH = 1:1:1 to 1:0:0 to 9:0: 1 (v / v / v)). The obtained methoxyethenyl intermediate was dissolved by adding THF (100 mL) and IN HCI (70 mL) was added. Then the mixture was stirred at rt for 2.5 h. The solution was ice-cooled and 2N NaOH (35 mL) was added thereto and the mixture was extracted with EtOAc. The organic layer was washed with brine and then dried over anhydrous Na2SO4. After filtration, the residue obtained by concentration under reduced pressure was purified by silica gel column chromatography (MeOH / EtOAc = 0:1 to 1:9 (v / v)) to obtain compound le as a colorless oily substance.TH NMR (CDCI3) 6 (ppm) 1.13-1.22 (m, 2H), 1.61-2.06 (m, 7H), 2.36 (dd, J = 6.6, 2.2 Hz, 2H), 2.57-2.64 (m, 1H), 3.61 (s, 3H), 5.55 (s, 2H), 6.98-6.99 (m, 1H), 7.12-7.14 (m, 1H), 8.33 (d, J = 5.4 Hz, 1H), 9.76 (t, J = 2.2 Hz, 1H).

[0338] Step 9: 2-(trans-4-(4-Methyl-5-(((4-(trifluoromethyl)pyridin-2-yl)oxy)methyl)-4 / 7-l,2,4- triazol-3-yl)cyclohexyl)acrylaldehyde (Id) Compound le (6.87 g) was dissolved in / V, / V-dimethylformamide (40 mL). To this, a formaldehyde solution (37%) (6.6 mL) and L-proline (618 mg) were added and the mixture was stirred at rt for 14 h. The mixture was diluted with EtOAc and washed with water and brine. The organic layer was dried over anhydrous Na2SO4. After filtration, the residue obtained by concentration under reduced pressure was purified by silica gel column chromatography (EtOAc / hexane / MeOH = 1:2:0 to 1:0:0 to 9:0:1 (v / v / v)) to obtain compound Id as a white solid.TH NMR (CDCI3) 6 (ppm) 1.34-1.44 (m, 2H), 1.85-1.98 (m, 4H), 2.03-2.08 (m, 2H), 2.56-2.63 (m, 1H), 2.64-2.71 (m, 1H), 3.62 (s, 3H), 5.55 (s, 2H), 5.98 (s, 1H), 6.25 (s, 1H), 6.99 (s, 1H), 7.13 (d, J = 5.4 Hz, 1H), 8.33 (d, J = 5.4 Hz, 1H), 9.52 (s, 1H).

[0339] Step 10: 2-(trans-4-(4-Methyl-5-(((4-(trifluoromethyl)pyridin-2-yl)oxy)methyl)-4 / 7-l,2,4- triazol-3-yl)cyclohexyl)oxirane-2-carbaldehyde (lc)

[0340] Compound Id (3.68 g) was dissolved in MeOH (60 mL). To this, a hydrogen peroxide solution (30%) (1.33 mL) and a 5N aqueous NaOH solution (0.47 mL) were sequentially added under ice-cooling and the mixture was stirred for 1 h. To the mixture, a 10% aqueous sodium thiosulfate solution was added and the reaction mixture was extracted with EtOAc. The organic layer was washed with brine, and then dried over anhydrous Na2SO4. The mixture was concentrated under reduced pressure, then CH2CI2 was added. The mixture was azeotropically concentrated to obtain compound lc as a white solid.1H NMR (CDCI3) 6 (ppm) 1.30-1.39 (m, 1H), 1.44-1.53 (m, 1H), 1.76-1.93 (m, 4H), 2.00-2.12 (m, 3H), 2.58-2.64 (m, 1H), 2.98 (d, J = 4.4 Hz, 1H), 3.10 (d, J = 4.4 Hz, 1H), 3.61 (s, 3H), 5.55 (s, 2H), 6.98-6.99 (m, 1H), 7.12-7.14 (m, 1H), 8.33 (d, J = 4.9 Hz, 1H), 8.85 (s, 1H).

[0341] Step 11: Benzyl (trans-4-(4-(trans-4-(4-methyl-5-(((4-(trifluoromethyl)pyridin-2- yl)oxy)methyl)-4 / 7-l,2,4-triazol-3-yl)cyclohexyl)-l / 7-pyrazol-l-yl)cyclohexyl)carbamate (lb)

[0342] Benzyl (trans-4-hydrozinylcyclohexyl)carbamate hydrochloride (442 mg) was suspended in EtOH (10 mL). To this, compound lc (504 mg) was added and the mixture stirred at rt for 43 h. The solvent was distilled away under reduced pressure and the resultant residue was purified by NH silica gel column chromatography (hexane / CH2Cb / EtOAc = 1:4:0 to 0:1:4 (v / v / v)). The resultant crude product was washed with EtOAc to afford compound lb as a white solid.TH NMR (CDCI3) 6 (ppm) 1.29-1.39 (m, 2H), 1.42-1.53 (m, 2H), 1.80-1.99 (m, 4H), 2.03-2.25 (m, 9H), 2.58-2.73 (m, 2H), 3.66 (s, 3H), 4.02-4.11 (m, 1H), 4.61-4.69 (m, 1H), 5.10 (s, 2H), 5.59 (s, 2H), 7.02 (s, 1H), 7.16 (d, J = 5.4 Hz, 1H), 7.22 (s, 1H), 7.30-7.39 (m, 6H), 8.36 (d, J = 5.4 Hz, 1H). Step 12: trans-4-(4-(trans-4-(4-Methyl-5-(((4-(trifluoromethyl)pyridin-2-yl)oxy)methyl)-4 / 7- l,2,4-triazol-3-yl)cyclohexyl)-l / 7-pyrazol-l-yl)cyclohexan-l-amine (la)

[0343] Compound lb (300 mg) was dissolved in meOH (6 mL) and CH2CI2 (1 mL). To this, acetic acid (0.14 mL) and 10% palladium on carbon (150 mg) were added at room temperature and the mixture was stirred at rt for 4 h under a H2 atmosphere. Then the catalyst was filtered off and the mixture was washed with CH2CI2. The resultant filtrate was concentrated under reduced pressure and the solvent was azeotropically concentrated with ethanol and toluene. The resultant residue was purified by silica gel column chromatography (MeOH / CH2Cb = 0:1 to 1:19 (v / v)) to obtain compound la as a white solid.TH NMR (CDCb) 6 (ppm) 1.25-1.35 (m, 2H), 1.43-1.54 (m, 2H), 1.75-2.24 (m, 14H), 2.59- 2.73 (m, 3H), 3.66 (s, 3H), 4.02-4.09 (m, 1H), 5.59 (s, 2H), 7.02 (s, 1H), 7.16 (d, J = 5.4 Hz, 1H), 7.23 (s, 1H), 7.38 (s, 1H), 8.36 (d, J = 5.4 Hz, 1H).

[0344] Step 13: 6-(trans-4-(4-(trans-4-(4-Methyl-5-(((4-(trifluoromethyl)pyridin-2-yl)oxy)methyl)-

[0345] 4 / 7-l,2,4-triazol-3-yl)cyclohexyl)-l / 7-pyrazol-l-yl)cyclohexyl)-l-oxa-6-azaspiro[3.3]heptane

[0346] (1)

[0347] Compound of oxetane-2,2-diyldimethanol (75 mg) and / V, / V-diisopropylethylamine (0.72 mL) were dissolved in CH2CI2 (1.5 mL). The mixture was cooled to -78°C, then trifluoromethanesulfonic acid anhydride (0.21 mL) was added dropwise thereto. The mixture was stirred at -78°C for 15 minutes. To this, a solution of compound la (213 mg) in CH2CI2 (5 mL) was added at 0°C and the mixture was stirred at 0°C for 15 minutes and then stirred at rt for 18 h. The residue obtained by concentration under reduced pressure was purified by NH silica gel column chromatography (MeOH / CI-hCb = 0:1 to 1:99 (v / v)). To the obtained crude product, diethyl ether was added, and the precipitated solid was collected by filtration to obtain title compound 1 as a white solid.1H NMR (CDCb) 6 (ppm) 1.16-1.26 (m, 2H), 1.42-1.53 (m, 2H), 1.70-1.80 (m, 2H), 1.89-2.19 (m, 11H), 2.58-2.72 (m, 2H), 2.88 (t, J = 7.5 Hz, 2H), 3.11- 3.16 (m, 2H), 3.64-3.68 (m, 2H), 3.65 (s, 3H), 3.99-4.07 (m, 1H), 4.53 (t, J = 7.5 Hz, 2H), 5.59 (s, 2H), 7.01-7.02 (m, 1H), 7.15-7.17 (m, 1H), 7.21 (s, 1H), 7.37 (s, 1H), 8.36 (d, J = 5.1 Hz, 1H). MS (APCI) m / z: 586 [M+H]+.

[0348] Example 2: Chemical synthesis of linkeable tool compound

[0349] Step 1: Benzyl 4-(4-(trans-4-(4-methyl-5-(((4-(trifluoromethyl)pyridin-2-yl)oxy)methyl)-4 / 7- l,2,4-triazol-3-yl)cyclohexyl)-l / 7-pyrazol-l-yl)piperidine-l-carboxylate (2b)

[0350] To a solution of compound lc (891 mg) in EtOH (8 mL) was added benzyl 4- hydrazineylpiperidine-l-carboxylate hydrochloride (546 mg). The mixture was stirred at rt for 16 h, concentrated and purified by reversed-phase chromatography (C18) (MeCN:0.1% NH4HCO3= 10 to 100%) to give compound 2b as a white solid. LCMS (ESI): m / z = 590.4 (M+H)+.

[0351] Step 2: 2-((4-Methyl-5-(trans-4-(l-(piperidin-4-yl)-l / 7-pyrazol-4-yl)cyclohexyl)-4 / 7-l,2,4- triazol-3-yl)methoxy)-4-(trifluoromethyl)pyridine (2a)

[0352] To a solution of compound lb (876 mg) in CH2CI2 (8 mL) was added TFA (1 mL). The mixture was stirred at rt for 2 h, concentrated and purified by reversed-phase chromatography (C18) (MeCN:0.1% NH4HCO3= 10 to 100%) to give compound 2a as a white solid.TH NMR (500 MHz, DMSO-d6) 6 8.50 (d, J = 5.0 Hz, 1H), 7.55 (s, 1H), 7.41 (d, J = 5.0 Hz, 1H), 7.32 (d, J = 4.5 Hz, 2H), 5.52 (s, 2H), 4.13-4.08 (m, 1H), 3.63 (s, 3H), 3.03 (d, J = 12.0 Hz, 2H), 2.84 (t, J = 12.0 Hz, 1H), 2.58 (t, J = 12.0 Hz, 2H), 2.52-2.51 (m, 1H), 2.04-1.90 (m, 6H), 1.78-1.64 (m, 4H), 1.51- 1.41 (m, 2H). LCMS (ESI): m / z = 245.8 (M / 2+H)+, 490.3 (M+H)+.

[0353] Step 3: 2-((4-Methyl-5-(trans-4-(l-(l-(2-(prop-2-yn-l-yloxy)ethyl)piperidin-4-yl)-l / 7-pyrazol- 4-yl)cyclohexyl)-4 / 7-l,2,4-triazol-3-yl)methoxy)-4-(trifluoromethyl)pyridine (2)

[0354] To a solution of compound 2a (114 mg) and 2-(prop-2-yn-l-yloxy)ethyl 4-methylbenzene- sulfonate (200 mg) in MeCN (8 mL) was added Nal (12 mg) and K2CO3 (170 mg). The mixture was stirred at 60°C for 16 h, cooled to rt, concentrated and purified by reversed-phase chromatography (C18) (MeCN:0.1% NH4HCO3 = 10 to 100%) to give compound 2 as a white solid.XH NMR (500 MHz, MeOD-d4) 6 8.42 (d, J = 5.0 Hz, 1H), 7.55 (s, 1H), 7.38 (s, 1H), 7.28 (d, J = 4.5 Hz, 1H), 7.18 (s, 1H), 5.61 (s, 2H), 4.18 (d, J = 2.5 Hz, 2H), 4.15-4.08 (m, 1H), 3.76 (s, 3H), 3.70 (t, J = 5.5 Hz, 2H), 3.13-3-11 (m, 2H), 2.94-2.85 (m, 2H), 2.67 (t, J = 5.5 Hz, 2H), 2.66- 2.59 (m, 1H), 2.31-2.25 (m, 2H), 2.14-1.99 (m, 8H), 1.83-1.75 (m, 2H), 1.61-1.52 (m, 2H). LCMS (ESI): m / z = 286.9 (M / 2+H)+, 572.3 (M+H)+.

[0355] Example 3: Content of different doses of compound 1

[0356] As described above, compound 1 can be dosed as free base or as a pharmaceutically acceptable salt and / or solvate thereof. The preferred polymorph of compound 1 (as free base) is described in WO2019 / 054427, i.e., is characterized by an X-ray powder diffraction pattern having characteristic peaks at 2 theta (±0.2°) of 13.2°, 15.8°, 16.5°, 17.8°, 18.1°, 20.3°, 20.8°, 21.4° and 27.9°. The quantity stated in the embodiments and claims relates to the amount of active moiety, i.e., free base. For optional salt and / or solvate the amount has to be adjusted. In the following table the amount (in mg) of active moiety of the compound is converted into pmol.

[0357] Example 4: Binding characterization to SIRT6 with compound 2 using surface plasmon resonance

[0358] Step 1: Immobilization of l-amino-ll-azido-3,6,9-trioxaundecane l-Amino-ll-azido-3,6,9-trioxaundecane was used to link the alkyne analogue of a compound according Formula (I) (compound 2) with the dextran matrix of the CM5 sensor chip. To exclude an unspecific binding of SIRT6 to this linker only the l-amino-ll-azido-3,6,9- trioxaundecane without compound 2 was coupled onto the sensor chip surface and a possible interaction with SIRT6 as analyte was tested.

[0359] • Running buffer: HBS-P (0.01 M HEPES pH 7.4, 0.15 M NaCI, 0.05% v / v Tween-20) + 1% (v / v) DMSO

[0360] • Activation surface with EDC / NHS (10 pL / min, 600 sec contact time)

[0361] • 100 pL (=110 mg) l-amino-ll-azido-3,6,9-trioxaundecane were taken from the original flask under argon atmosphere and transferred into an argon rinsed 0.5 mL tube

[0362] • Dilution in 10 mM sodium phosphate pH 6.5: 990 pL sodium phosphate pH 6.5 + 9.09 pL l-amino-ll-azido-3,6,9-trioxaundecane 10 mg / mL

[0363] • Injection of 10 mg / mL l-amino-ll-azido-3,6,9-trioxaundecane: Flow rate: 10 pL / min

[0364] Injection of 300 pL of 10 mg / mL solution, then flow rate: 5 pL / min Injection of 325 pL of 10 mg / mL solution

[0365] • Deactivation of sensor chip surface: Flow rate: 10 pL / min, injection of 100 pL 1 M ethanolamine (contact time 600 sec)

[0366] • Rinsing the sensor chip overnight with 10 pL / min running buffer A response increase of ~480 to ~590 resonance units (RU) was obtained (depending on the experiment) with this linker element coupled to the dextran matrix.

[0367] Step 2: Immobilization of compound 2 with amine-azide-linker via Copper-catalysed click chemistry onto a CM5 chip

[0368] • Dissolving of compound 2 in DMSO to a concentration of 50 mM

[0369] • Dissolving of 0.49 g sodium ascorbate in 4.9 mL water 504.77 mM

[0370] • Dissolving of 0.49 g CuSC -SI-hO in 4.711 mL water 416.51 mM

[0371] • Dilution of sodium ascorbate down to 50.477 mM in HBS-P

[0372] • Dilution of CuSC down to 4.1651 mM in HBS-P

[0373] • Pipetting of 50 pL 50 mM compound 2 into a 2 mL tube

[0374] • Fast Addition of 933 pL HBS-P and fast mixing to prevent precipitation of compound 2

[0375] • Addition of 4.95 pL 50.4 mM sodium ascorbate

[0376] • Addition of 12 pL 4.16 mM CuSC

[0377] • Mixing again 2.5 mM MWT-S-01343 in HBS-P with 5% DMSO

[0378] • Flow rate down to 2 pL / min

[0379] • Injection of 325 pL 2.5 mM compound 2 in HBS-P with 5% DMSO

[0380] • Injection of 170 pL 2.5 mM compound 2 in HBS-P with 5% DMSO

[0381] • Rinsing the sensor chip overnight with 10 pL / min running buffer

[0382] Coupling of compound 2 via the linker onto chip surface gave an increase of ~20 RU (the usage of 5% DMSO (according CM5 chip specification a 1-minute pulse with max. 10% DMSO is emphasized) has possibly led to washing out the dextran matrix during the coupling). The successful coupling was proven in the next step by SIRT6 binding.

[0383] Step 3: Interaction analysis between compound 2 and SIRT6 (without NAD)

[0384] A control experiment with linker alone (without coupled compound 2) bound to the chip surface revealed, that injection of SIRT6 (without or with 500 pM NAD) can be performed without organic solvents (DMSO, methanol) minimizing bulk effects and resulted in a good and stable baseline without a strong drift. Only a weak unspecific interaction of SIRT6 with the amine-azide linker was observed. Now the interaction of SIRT6 to compound 2 bound to the chip was analyzed in absence and presence of NAD. This measurement was performed with following running setup:

[0385] • Running buffer: 20 mM Tris, 150 mM NaCI, 0.05% Tween-20, 1 mM DTT (freshly dissolved) pH 8.0

[0386] • SIRT6 (1 mg / mL equal to 27.5 pM), stored in 150 mM NaCI, 20 mM Tris, 0,5 mM TCEP pH 7.6 at -80°C was thawed and following dilutions concentrations were generated by serial dilutions in running buffer: 2187 nM, 729 nM, 243 nM, 81 nM, 27 nM, 9 nM, 3 nM and 1 nM

[0387] • Starting the Sensorgram recording • Flowpath: 1,2; Detection: 2-1; Flow: 30 pL / min

[0388] • Kinject: 240 pL (contact time: 480 sec), 600 sec dissociation time

[0389] • Injections: 1 nM SIRT6, 3 nM SIRT6, 9 nM SIRT6, 27 nM SIRT6, 81 nM SIRT6, 243 nM SIRT6, 729 nM SIRT6 and finally 2187 nM SIRT6

[0390] The resulting sensorgram is shown in Figure 1A. The sensorgram shows a very good and stable baseline. Only weak signals were observed which were evaluated by steady-state analysis obtaining a KD value of 2.16 pM, however, due to the weak curvage of the fit the calculated constants are error-prone. This finding indicates that in absence of NAD the interaction of SIRT6 to compound 2 is weak.

[0391] Step 4: Interaction analysis between compound 2 and SIRT6 (with NAD)

[0392] Afterwards the interaction was analyzed in presence of NAD. This binding experiment was performed with following running setup:

[0393] • Running buffer: 20 mM Tris, 150 mM NaCI, 0.05% Tween-20, 1 mM DTT (freshly dissolved) pH 8.0

[0394] • To 4 mL running buffer 500 pM NAD was added

[0395] • SIRT6 (1 mg / mL equal to 27.5 pM), stored in 150 mM NaCI, 20 mM Tris, 0,5 mM TCEP pH 7.6 at -80°C was thawed and following dilutions concentrations were generated by serial dilutions in running buffer with 500 pM NAD: 2187 nM, 729 nM, 243 nM, 81 nM, 27 nM, 9 nM, 3 nM andl nM

[0396] • Starting the Sensorgram recording

[0397] • Flowpath: 1,2, 3, 4; Detection: 2-1, 3-1, 4-1; Flow: 30 pL / min

[0398] • Kinject: 240 pL (contact time: 480 sec), 300 sec dissociation time

[0399] • Injections: running buffer with 500 pM NAD: 1 nM SIRT6, 3 nM SIRT6, 9 nM SIRT6, 27 nM SIRT6, 81 nM SIRT6, 243 nM SIRT6, 729 nM SIRT6 and finally 2187 nM SIRT6

[0400] The resulting sensorgram is shown in Figure 2A. Control experiments indicate that the binding of SIRT6 is strongly mediated by the co-substrate NAD. The evaluation of the binding data shows a very strong interaction of SIRT6 and compound 2 in presence of NAD with a KD value of 99 pM, which is mainly driven by a very slow off-rate (kd):

[0401] After analyzing the interaction, the chip surface with immobilized compound 2 was initially regenerated with mobile compound 1 to compete the binding of SIRT6 to compound 2, however this approach failed, which indicated that the complex of compound 2 and NAD is strongly anchored in the active site of SIRT6 that compound 1 alone cannot displace it. This indicates that also the binding of compound 1 to SIRT6 requires NAD, otherwise the replacement should occur. Conclusions:

[0402] • SIRT6 and compound 2 (an analogue representing Formula (I) and Formula (II)) can interact only in presence of NAD very tightly.

[0403] • SIRT6 catalyzes the transfer of the ADP moiety of NAD to compound 2 and forms a covalent conjugate that blocks the active site of SIRT6 very tightly.

[0404] • This binding mode is unique to SIRT6 and was confirmed by X-ray crystallography of a compound fo Formula (I) to SIRT6.

[0405] Example 5: Selectivity of compound 1 for SIRT6 over other members of the sirtuin family

[0406] Inhibition of the enzymatic activity mediated by compound 1 is highly selective for SIRT6 over other members of the human sirtuin protein family as well as members of other classes of histone deacetylases (HDACs). Compound 1 did not inhibit the enzyme activities of other tested sirtuin family members (IC50 >1000 nM for hSIRTl, hSIRT2, hSIRT3, hSIRT5 and hSI RT7) and compound 1 was determined to be highly selective for SIRT6 over other SIRT family members (Figure 2).

[0407] The modulatory potential of compound 1 on SIRT6 was assessed in vitro using an AlphaLISA immunodetection assay. In a cell free system, the half-maximal inhibitory concentration (IC50) was 15 nM for the human SIRT6. IC50 values for mouse, monkey and rat SIRT6 were 41 nM, 31 nM and 28 nM, respectively.

[0408] Besides deacetylation, compound 1 effectively inhibited SIRT6's deacylase activity (demyristoylation of H3K9) with an IC50 value of 41 nM in vitro. Modulation of SIRT6 was also tested in a cellular system in HT-29 cells. The half-maximum effectivity concentration (EC50) in these cells was 4.3 nM. In addition, treatment of Caco-2 cells and human intestinal organoids with compound 1 stabilizes SIRT6 protein and increases SIRT6 protein levels.

[0409] Experimental details of the in vitro enzymatic sirtuin activity assay shown in Figure 2:

[0410] The effects of compound 1 on the deacetylase activities of human sirtuin family members SIRT6, SIRT1, SIRT2, SIRT3, and SIRT7 as well as the desuccinylase activity of human SIRT5 were measured using an AlphaLISA immunodetection assay.

[0411] In detail, recombinant sirtuin proteins, biotinylated peptides, and compound 1 or the respective vehicle (1% DMSO) were incubated with or without p-nicotinamide adenine dinucleotide (NAD+, Sigma-Aldrich Co. LLC.) in 10 pL assay buffer [10 mM Tris-HCI pH 8.0 (Wako Pure Chemical Industries, Ltd.), 0.1% (w / v) bovine serum albumin (Sigma-Aldrich Co. LLC.), 0.01% Tween 20 (Bio-Rad Laboratories), 1 mM dithiothreitol solution (DTT, Sigma- Aldrich Co. LLC.), and 12.5% glycerol (Wako Pure Chemical Industries, Ltd.)] in a 384-well Alpha plate (PerkinElmer Inc.) at rt for 1 h. After the enzymatic reactions, 5 pL of AlphaLISA acceptor beads mixture [40 pg / mL AlphaLISA Protein A Acceptor beads (PerkinElmer Inc.), detection antibodies for each substrate and 8 mM nicotinamide (NAM, Sigma-Aldrich Co. LLC.) in AlphaLISA Epigenetics Buffer (PerkinElmer Inc.)] were added and incubated at rt for 1 h. Thereafter, 5 pL of 40 pg / mL AlphaScreen Streptavidin Donor beads (PerkinElmer Inc.) in Alpha LISA Epigenetic Buffer were added and incubated at rt for 30 min, shielded from light. The AlphaLISA signal was measured on the EnVision Multilabel Plate Reader (PerkinElmer Inc.). The values for enzyme activity for each sample were calculated using Microsoft Excel as below:

[0412] • Enzyme activity (%) = 100 - 100 x (Signal in the presence of NAD+) - (Signal in the absence of NAD+)

[0413] • Inhibition (%) = 100 - 100 x (Enzyme activity of test sample) / (Enzyme activity of vehicle control)

[0414] The 50% inhibitory concentration (IC50) values were determined using SAS system Release 9.2 software (SAS Institute, Inc.) according to the Sigmoid Emax model as follows:

[0415] • Inhibition (%) = [(Emax- Eo) x O'] / [(logi0IC5o)v_+ ] + Eo

[0416] • C: Common logarithmic concentration of the compound (nM)

[0417] • y: Sigmoidicity factor

[0418] Emax and Eo were fixed at 100% and 0%, respectively.

[0419] The table below summarizes the conditions of the enzymatic activity assay used for evaluating selectivity of compound 1 towards other sirtuin family members. sirtuin protein NAD+ Peptide

[0420] Detection antibody

[0421] (pg / mL) (pM) (nM)

[0422] SIRT6 0.1 15 H3K9ac 1 Anti-H3K9ac 0.5 pg / mL

[0423] SIRT1 0.01 15 H3K9ac 1 Anti-H3K9ac 0.5 pg / mL

[0424] SIRT2 2 50 H3K18ac 3 Anti-H3K18ac 0.1%

[0425] SIRT3 0.1 50 LCAD_K318ac 100 Anti-AcK 0.1%

[0426] SIRT5 0.1 15 CPSl_K1297suc 3 Anti-SucK 0.1%

[0427] SIRT7 0.3 100 H3K18ac 3 Anti-H3K18ac 0.1%

[0428] Conclusion:

[0429] Compound 1 is a potent SIRT6 inhibitor with high selectivity for SIRT6 over other human sirtuin family proteins.

[0430] Example 6: Selectivity of reference compounds on SIRT6 and other members of the sirtuin family

[0431] Notably, SIRT6 inhibitors described in the literature often lack the high selectivity and potency compared to compound 1, and exert substantial inhibitory effects on other sirtuin members, especially SIRT1, SIRT2 and SIRT3, or their selectivity towards other sirtuin members has not been tested so far (Fiorentino et al. J. Med. Chem. 2021;64:9732). Reference compounds can be tested, e.g. in the SIRT6 activity assay kit from Abeam Ltd, which detects deacetylase activity of recombinant human SIRT6 (Fluorometric, catalogue# abl56068). The assay is also available for SIRT1, SIRT2 and SIRT3. The literature gives hints that modulation of SIRT6 might be involved in GvHD, even though it's unclear, which modulation, either on protein stability and interaction or on enzymatic activity is responsible for the effect. For example, WO2014 / 170873 and accompaning publication Bioorg. Med. Chem. 2018;25:5849 describes commercially available screening compounds as SIRT6 inhibitors with ICso-values in the high micromolar range and mentions GvHD within a broad range of diseases with no enablement in this direction. Surprisingly, compound 1 seems to have the perfect and unique combination of affecting SIRT6 protein and function.

[0432] Example 7: Selectivity of compound 1 in the SafetyScreen87™

[0433] In vitro pharmacological profiling was performed with compound 1 against a broad range of targets (receptors, transporters, ion channels, and enzymes) in the SafetyScreen87™ (Eurofins Discovery) to screen for potential undesirable molecular interactions that may cause adverse drug reactions in humans. Compound 1 was tested, in duplicate, at a concentration of 10 pM against this panel of 87 targets. Test compound binding was calculated as a percentage inhibition of the binding of a radioactively labelled ligand specific for each target. Test compound enzyme inhibition effect was calculated as a percentage inhibition of control enzyme activity. Inhibition or stimulation higher than 50% represented a response that was considered significant. No significant results were noted with responses to all targets less than 40%.

[0434] Example 8: In vitro intestinal barrier function model

[0435] Caco-2 cell line is derived from human epithelial cells from the colon. They are frequently used to mimic intestinal barrier function to either look at permeability of drugs or to look at is functional resistance. For the present experiment, the Caco-2 cells were used to mimic the loss of its barrier function upon challenging the cells with 2,4,6-trinitrobenzene sulfonic acid (TNBS) and investigate the effect of compound 1 on this by measuring the electrical transepithelial resistance (TEER). TNBS is a chemical that in in vivo models is often used to induce a disease that shows similarity's with Crohn's disease in humans. For this Caco-2 cells, cultivated in a 6-well trans well plate on top of the membrane that has small pores that prevent the cells from going through but allow molecules and nutrients to pass. After 3 weeks, when the TEER is stable, the monolayer is challenged with 0.25% TNBS stimulation for 4 h. Then, cells were washed and fresh medium with vehicle or compound 1 was given. TEER was measured before and after challenge at different time points. For correct measurement, the shorter electrode is inserted into the transwell chamber (without touching the cells), and the longer electrode is inserted in the outer well.

[0436] The results show a higher TEER and a higher recovery rate for cells treated with compound 1 compared to vehicle control. This indicate that treatment with compound 1 induces a tightening of the barrier and that therefore, compound 1 can be active in diseases that show a reduced intestinal barrier function like in GvHD. Example 9: Therapeutic dextran sodium sulfate (DSS)-induced colitis mouse model

[0437] For this model, which is an often-used in vivo model for IBD, male C57BL / 6 mice were treated with 2.8% DSS in drinking water for 5 days. Treatment with either vehicle (PEG400), 60 mg / kg sulfasalazine (positive control) or 1 mg / kg of compound 1 was given at disease onset (day 4 after start of DSS treatment) per oral gavage once daily for 5 days. Diarrhea score (normal consistency; soft; pasty, unshaped, not sticking to anus; diarrhea, sticking to anus; diarrhea with macroscopic bleeding) was assessed daily. At the end of the study, mice were sacrificed and the colon was collected. Colon histology score consisting of an architecture score (0-3) and an infiltration score (0-3) was assessed based on hematoxylin and eosin-stained tissues slices.

[0438] This model showed that drinking of DSS induced an upregulation of the diarrhea score and this was also reflected in an increased histological score. The positive control showed a reduction in the diarrhea score while no improvement on the histology score was observed. Compound 1 also showed an improvement of the diarrhea score which was similar or even better than the positive control. In addition, compound 1 was able to also show a reduction in the histology score.

[0439] These results show that compound 1 has activity in the DSS-colitis model, which indicates its potential activity in human diseases in which a defective intestinal barrier function and intestinal inflammation play an important role such as in GvHD.

[0440] Example 10: In vitro activity screen in a hematologic tumor cell line panel

[0441] Many patients who receive an allogeneic hematopoietic stem cell transplant and develop or are at risk of developing GvDH suffer from haematological malignancies such as leukemia or lymphoma.

[0442] To determine any anti-cancer or tumor promoting activity mediated by SIRT6 modulation in hematologic cancers, effects of compound 1 on cell viability were assessed in a panel of 58 hematologic cell lines using a CellTiter-Glo® cell viability assays. The CellTiter-Glo® luminescent cell viability assay is a widely used screening method in anti-cancer drug development and determines the number of viable cells based on quantitation of adenosine triphosphate (a marker of metabolically active cells) present in the culture.

[0443] In this assay compound 1 was tested at 10 concentrations in half-log increments (highest concentration 30 pM) and added to cells 24 h after cell seeding. After 72 hour-long incubation of cells with compound 1, cells were lysed and incubated in CellTiter-Glo® One Solution Assay reagent followed by luminescence reading. IC50 values were calculated for each tested tumor cell line. Anti-tumor activity and potential tumor promoting activity is expressed by test / control (T / C) values at each test concentration and in case of growth inhibition by IC50 values calculated by 4 parameter non-linear regression analysis.

[0444] In most of the cell lines compound 1 showed no or only marginal antitumor activity across the panel of 58 hematological cell lines. Significant inhibition of growth (T / C <50%) was onlyfound in individual cell lines (EOL-1, MEC-1, KM-H2, DOHH-2, HH, LP-1) at the highest test concentration of 30 pM. The highest activity was detected for the CLL cell line MEC-1 (abs. IC50 of 10.7 pM) and NHL cell line DOHH-2 (14.9 pM). In some other cell lines compound 1 showed marginal activity (T / C between 50% and 70%) at the highest test concentration of 30 pM, but in most cell lines (32 / 58) a T / C <70% was not achieved at any test concentration. Summary: Potential tumor promoting activity, as expressed by stimulation of growth (increasing T / C values) was not indicated in any cell line for compound 1. SIRT6 modulation by compound 1 did neither show prominent anti tumor activity, nor potential tumor promoting activity.

[0445] Example 11: In vivo GvHD mouse model

[0446] Acute GvHD can be modeled in vivo using a MHC-mismatch C57BL / 6 BALB / c mouse model. BALB / c recipient mice were pre-conditioned with a lethal dose of total body irradiation (day 1) prior to the adoptive transfer of T-cell depleted bone marrow supplemented with splenocytes harvested from a C57BL / 6 donor mouse (day 2). Following transplant, the mice develop diarrhea and lose weight due to preconditioning. Then partially recover by day 14, after which they develop a progressively worsening disease (weight loss and increasing GvDH score).

[0447] Prophylactic treatment with compound 1 (3 mg / kg per day, p.o.) was started from day 0 until day 56 and compared to vehicle treated animals (0.5% methylcellulose, p.o).

[0448] Parameters to assess disease severity: survival body weight

[0449] GvDH score

[0450] Plasma / serum cytokines

[0451] Intestinal inflammation assessed by measuring: endoscopy score (video endoscopy of colon and scoring of disease severity) diarrhea score colon weight / length ratio

[0452] - jejunum weight / length ratio

[0453] H&E staining of intestinal tissue section to assess mucosal damage RNAseq analysis of RNA isolated from small and large intestinal tissue

[0454] Example 12: Proof-of-concept trial for prevention of acute GvHD following allogeneic HSCT comparing compound 1 versus current standard-of-care

Claims

CLAIMS1. A compound of Formula (I)Formula (I), or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft -versus- host disease in a subject in need thereof, wherein:R1is selected from the group consisting of hydrogen, carboxyl, cyano, fluorine, chlorine, methyl, isopropyl, t-butyl, trifluoromethyl, trifluoromethoxy, cyclopropylmethoxy, 1,1- difluoro-2-methylpropyl, l,l-difluoro-2,2-dimethylpropyl, 1-methyl-l-cyclobutyl, methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl, 1-hydroxy-l-methylethyl, azetidine-l-carbonyl, 3-methyloxetan-3-yl, 4,5-dihydrooxazol-2-yl and cyclopropylcarbonyl;R2is hydrogen or methyl;A1is =N- or =CH-;X is benzene, pyridine or cyclohexane;J is any ring selected from the groupY is selected from the group consisting of phenyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl,piperidinyl, azetidinyl, tetra hydropyranyl, morpholinyl and tetrahydropyridinyl, optionally substituted with 1, 2 or 3 groups selected from Yl, or alternatively Y is amino, optionally substituted with 1 or 2 groups selected from Yl;Y1is selected from the group consisting of hydroxy, cyano, fluorine, amino, methyl, ethyl, isopropyl, methoxy, azetidinyl, pyrrolidinyl, morpholinyl,, wherein amino is optionally substituted with 1 or 2 groups selected from Y2and methyl, ethyl, isopropyl, methoxy, azetidinyl, pyrrolidinyl, morpholinyl,are optionally substituted with 1, 2 or 3 groups selected from Y2;Y2is selected from the group consisting of hydroxyl, fluorine, amino, methyl, methoxy, azetidinyl, pyrrolidinyl, morpholinyl,wherein amino is optionally substituted with 1 or 2 groups selected from Y3and methyl, methoxy, azetidinyl, pyrrolidinyl, morpholinyl,is optionally substituted with 1, 2 or 3 groups selected from Y3,Y3is hydroxyl, fluorine, cyano, methyl or methoxy, wherein methyl or methoxy are optionally substituted with fluorine.

2. A compound of Formula (I) or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof according to claim 1, wherein the compound is selected from the following group:A compound of Formula (I) or an isotopic variant, a pharmacologically acceptable salt thereof ora solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof according to claim 1 or 2, wherein the compound isFormula (II).

4. The compound of Formula (II), which isfor use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof.

5. A compound of Formula (I) or an isotopic variant, a pharmacologically acceptable salt thereof ora solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft -versus-host disease in a subject in need thereof according to any of claims 1 to 4, wherein said subject is primarily affected or at risk to be primarily affected in the gastrointestinal tract with symptoms like severe diarrhea or abdominal pain.

6. A compound of Formula (I) or an isotopic variant, a pharmacologically acceptable salt thereof ora solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft -versus-host disease in a subject in need thereof according to any of claims 1 to 5, wherein said subject suffers from a disease selected from from malignant disorders (such as acute myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, myelodysplastic syndromes, myeloproliferative disorders, nonHodgkin's lymphoma, Hodgkin's disease, chronic lymphocytic leukemia, multiple myeloma, juvenile chronic myeloid leukemia) or non-malignant disorders (such as aplastic anemia, paroxysmal nocturnal hemoglobinuria, Fanconi's anemia, Blackfan- Diamond anemia, sickle cell anemia, thalassemia major, severe combined immunodeficiency, Wiskott-Aldrich syndrome, inborn errors of metabolism).

7. A compound of Formula (I) or an isotopic variant, a pharmacologically acceptable salt thereof ora solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft -versus-host disease in a subject in need thereof according to any of claims 1 to 6, wherein said subject has a planned conditioning regimen in the context of an allogeneic HSCT.

8. A compound of Formula (I) or an isotopic variant, a pharmacologically acceptable salt thereof ora solvate thereof or a solvate of a salt thereof for use in a method of preventionor treatment of graft-versus-host disease in a subject in need thereof according to any of claims 1 to 7, wherein the subject is selected based on higher levels of plasma regenerating islet-derived 3 alpha (REG3a).

9. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof ora solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft -versus-host disease in a subject in need thereof according to any of claims 1 to 8, wherein the graft-versus-host disease is selected from chronic GvHD (cGvHD), acute GvHD (aGvHD), gastrointestinal GvHD, steroid-refractory GvHD, steroid- resistant GvHD or transfusion-associated graft-versus-host disease.

10. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof ora solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof according to any of claims 1 to 9, wherein said compound or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject in a daily dose of 34 pmol to 1370 pmol.

11. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof ora solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof according to any of claims 1 to 10, wherein said compound or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject with at least one additional therapeutic treatment.

12. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof ora solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof according to any of claim 11, wherein said at least one additional therapeutic treatment is an immunosupressive treatment.

13. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof ora solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof according to claim 12, wherein said immunosupressive treatment is selected from the group consisting of prednisone, prednisolone, methylprednisolone, budesonide, beclomethasone, dexamethasone, tacrolimus, sirolimus (rapamycin), cyclosporine, rifaximin, acalabrutinib, anlotinib, baricitinib, bortezomib, carfilzomib, entospletinib, fostamatinib, glasdegib, ibruitinib, imatinib, itacitinib, ixazomib (MLN9708), jaktinib, nilotinib, nintedanib, pacritinib, pimicotinib, ruxolitinib, upadacitinib, sonidegib, vismodegib, other JAKinhibitors (e.g. SHR0302), axatilimab, alemtuzumab, basiliximab, belimumab, brentuximab vedotin, daclizumab, efalizumab, gavilimomab (ABX-CBL), ibritumomab tiuxetan, infliximab, inolimomab, itolizumab, milatuzumab, muromonab-CD3, natalizumab, neihulizumab, obinutuzumab, ofatumumab, rituximab, siplizumab, tildrakizumab, tocilizumab, ustekinumab, vedolizumab, visilizumab, etanercept, alefacept, abatacept, IL-2, teduglutide, lenalidomide, pomalidomide, leflunomide, thalidomide, panobinostat (LBH589), alvelestat (MPH966), vorinostat, mycophenolate mofetil (MMF), methotrexate (MTX), decitabine, clofarabine, melphalan, thiotepa, pentostatin, palifermin, filgrastim, prochymal (remestemcel-L), cyclophosphamide, defibrotide, cannabidiol, VM-001 (from ViGenCell), RGI-2001, ASC930, TQ05105 (or other JAK / ROCK inhibitors), GDC-8264, ALPN-101, voriconazole, itraconazole, opebacan, belumosudil (KD025), efmarodocokin alfa, efavaleukin alfa (AMG 592), efprezimod alfa, romidepsin, busulfan, fludarabine phosphate, aldesleukin, thymoglobulin, methoxsalen, extracorporeal photopheresis (ECP), mesenchymal stromal cells (e.g. CYP-001, OTI-OIO or MC0518), allogeneic faecal microbiota (e.g. MaaT013), fecal microbiota trasnplantation, BET inhibitor (e.g. PLX51107), C5a inhibitor (e.g. ALXN1007), sitagliptin, atorvastatin, clobetasol, maraviroc, ribaxamase (SYN-004), sargramostim, alpha 1- antitrypsin, mitoxantrone or combinations thereof.

14. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof according to claim 13, wherein said agent is selected from prednisone, prednisolone, methylprednisolone, budesonide, beclomethasone, dexamethasone, ruxolitinib, upadacitinib, belumosudil or combinations thereof.

15. A compound of Formula (I) an isotopic variant, or a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof for use in a method of prevention or treatment of graft-versus-host disease in a subject in need thereof according to any of claims 1 to 14, wherein said compound or an isotopic variant, a pharmacologically acceptable salt thereof or a solvate thereof or a solvate of a salt thereof is administered to said subject for a period of at least 4 weeks, preferably at least 14 weeks, more preferably at least 26 weeks post HSCT.

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