Substituted fused imidazole derivatives and methods of treating sickle cell disease and related complications

Abstract

The present invention provides methods of treating sickle cell disease and related complications using compounds of Formula (I) alone or in combination with other active agents, and pharmaceutical compositions thereof.

Description

TECHNICAL FIELD

[0001] The present invention provides methods of using compounds of Formula (I) and pharmaceutical compositions thereof, alone or in combination with other active agents, to treat sickle cell disease and related complications. The present invention also provides compounds and pharmaceutical compositions. BACKGROUND

[0003] Sickle cell disease (SCD) is a life-threatening monogenic disease. SCD is a severe hemoglobinopathy that produces multisystem complications due to the expression of abnormal sickle hemoglobin (HbS). The most common type of SCD is sickle cell anemia (SCA) (also known as HbSS or SS disease or hemoglobin S), in which the mutation leading to HbS is homozygous. Less common types of SCD include sickle hemoglobin C (HbSC), sickle beta + thalassemia (HbS / β + ) and sickle beta0thalassemia (HbS / β 0 ).

[0004] Sickle cell disease (SCD) is caused by a point mutation that leads to deformed or sickle-shaped red blood cells. Sickle-shaped red blood cells are associated with clinical manifestations of SCD, such as anemia, recurrent painful vaso-occlusive crises, infections, acute chest syndrome, pulmonary hypertension, stroke, priapism, bone necrosis, renal insufficiency, leg ulcers, retinopathy, and heart disease.

[0005] SCD is caused by a single point mutation in codon 6 of the HBB globin gene (GAG > GTG). Hypoxic venous circulation leads to a self-assembly (polymerization) process that produces sickle hemoglobin molecules (HbS) and disrupts the membrane and cytoskeleton of red blood cells. HbS repeatedly enters the sickle and non-sickle cycle, progressively increasing damage to the red blood cell membrane (ischemia-reperfusion (IR) injury), leading to irreversibly sickled red blood cells. As a result, these rigid blood cells are unable to deform as they pass through narrow capillaries, leading to vaso-occlusion and ischemia. Actual anemia in this disease is caused by hemolysis, the destruction of red blood cells caused by their deformity.

[0006] C-reactive protein (CRP) and markers of oxidative stress are significantly increased after IR injury. Secondary oxidative stress leads to hemolysis, inactivation of nitric oxide (NO), and adhesive properties of red blood cells, white blood cells, and platelets.

[0007] Sickle-shaped red blood cells participate in a multi-step vaso-occlusive process with endothelial cells, activated white blood cells, platelets, and plasma proteins.

[0008] Heme oxygenase-1 (HO-1) and interleukin 10 (IL-10) are increased in SCD patients in an attempt to counteract the induced inflammation. HO-1 breaks down heme released during hemolysis, thereby limiting oxidative stress and inflammation, while IL-10 limits the production of proinflammatory cytokines.

[0009] Sickle red cells stimulate leukocyte recruitment: Following a secondary inflammatory stimulus, leukocytes are recruited to the activated endothelium of the venous circulation, where it forms adhesive interactions with the activated endothelium and sickle red cells, leading to reduced blood flow and ultimately to vessel occlusion.

[0010] SCD platelets show increased surface expression of selectin P (SELP), activated αIIb P-Selectin (SELP), activated

[0011] Hydroxyurea (HU) is an approved treatment that alters the disease course of SCD. HU counteracts the pathophysiology of SCD by increasing the production of red blood cells containing fetal hemoglobin (HbF) and indirectly altering gene expression and proteins associated with the pathophysiology of SCD. The increased concentration of HbF-containing red blood cells dilutes the concentration of sickle red cells, which in turn can trigger a cascade of reduced hemolysis, increased bioavailability of NO, and reduced endothelial activation. However, HU has been shown to reduce the white blood cell count in patients receiving treatment. Although HU improves clinical symptoms by reducing pain and vaso-occlusive crises, acute chest syndrome, transfusion requirements, and hospitalization, SCD patients receiving HU treatment have exhibited side effects such as induction of DNA damage, reduced sperm count, and production of iron-nitrosyl hemoglobin.

[0012] Thus, there is a need in the art for new, improved, and / or supplemental therapies for SCD. SUMMARY

[0013] PCT Publication No. WO 2011 / 103018 ("WO'018") describes substituted fused imidazole derivatives that upregulate HMOX1 expression in vitro. PCT Publication No. WO 2012 / 094580 ("WO'580") describes various compounds that modulate cellular oxidative stress, including fused imidazole derivatives having similar or identical structures to the compounds disclosed in WO'018.

[0014] The present invention relates to methods and compositions related to the treatment of one or more blood disorders. Although the blood disorder is SCD in particular embodiments, in specific embodiments, one or more other blood disorders can be treated with the present invention: a bleeding disorder (e.g., including a coagulopathy, a hypercoagulable state, hemophilia, or von Willebrand disease), a platelet disorder (e.g., essential thrombocythemia or thrombocytopenia), and / or hemophilia or anemia can be treated. In particular embodiments of the present invention, there are methods and compositions for treating and / or preventing sickle cell disease, which can be referred to as sickle cell anemia (or anemia; SCA) or sickle cell anemia.

[0015] Mammals and / or non-human mammals or cell lines can be used as a model for sickle cell disease. In particular cases, the individual being treated with the methods and / or compositions of the present invention can be experiencing a vaso-occlusive crisis, an acute chest crisis, a chest pain syndrome, which can or can not require hospitalization. In specific embodiments, the individual can be experiencing or can experience negative side effects of a drug, e.g., a drug that directly or indirectly causes increased coagulation and / or increased inflammation; in specific embodiments, the drug is HU.

[0016] In certain embodiments of the present invention, the compounds of the present invention are administered alone. In other embodiments, the compounds of the present invention are administered with one or more other drugs, some of which can or can not induce HbF production, to treat SCD. For example, the compounds of the present invention can be administered in combination with HU to treat SCD. In another example, the compounds of the present invention can be administered in combination with Nrf2 activators, such as fumarates (MMF or DMF) and bardoxolone methyl.

[0017] The individual receiving treatment can be known to have SCD, suspected of having SCD, or at risk for SCD. In embodiments of the present invention, the individual is diagnosed with sickle cell disease prior to receiving treatment of the present invention.

[0018] The present invention also relates to compounds of Formula (I) and pharmaceutically acceptable salts thereof and pharmaceutical compositions comprising Formula (I) and pharmaceutically acceptable salts thereof, and methods of making the same. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figures 1A-1D Relative cell growth and percentage of cell viability of KU812 cells in the presence of different concentrations (0, 0.5, 2.5, 5, 10, and 20 mM) of compounds 73, 134, 473, and 236 are shown, respectively. Data are presented as mean ± SD (n = 3). *, p < 0.05

[0020] Figure 1EWestern blot analysis was included, showing the HbF induction levels after treatment of KU812 cells with different concentrations (0, 0.5, 2.5, 5, 10, and 20 μM) of compounds 73, 134, 473, and 236. Hydroxyurea (HU) and heme were used as positive controls for HbF induction, and β-actin was used as an internal protein control.

[0021] Figure 2 The HbF protein expression level of KU812 cells obtained by FAC is shown and analyzed as the average HbF concentration per cell as measured by mean fluorescence intensity (MFI).

[0022] Figure 3A This includes Western blot analysis, which shows the induction levels of HbF and HbS in sickle erythroid progenitor cells after treatment with compound 473 (0.5 and 2.5 μM) for 48 hours. Hydroxyurea (HU) and heme chloride were used as positive controls for HbF induction, and β-actin was used as an internal protein control.

[0023] Figure 3B The percentage of HbF-positive cells (F cells) when sickle erythroid progenitor cells were treated with compound 473 (0.5 and 2.5 μM) for 48 hours and analyzed by flow cytometry is shown. Hydroxyurea (HU) and heme chloride were used as HbF-inducing positive controls.

[0024] Figure 4A Images of sickle erythroid progenitor cells included, which were cultured for 10 days and then treated with compound 473 at concentrations of 0.5 μM and 2.5 μM for 48 hours or with heme (approximately 50 μM) or hydroxyurea (HU) (approximately 100 μM), and then subjected to hypoxic conditions (1% O2 and 5% CO2).

[0025] Figure 4B The percentage of sickle cells is shown when sickle erythroid progenitor cells are cultured for 10 days and then treated with compound 473 at concentrations of 0.5 μM and 2.5 μM for 48 hours, or with heme (approximately 50 μN) or hydroxyurea (HU) (approximately 100 μM), and then subjected to hypoxic conditions (1% O2 and 5% CO2). Detailed Implementation

[0026] I. Definition

[0027] The following definitions are intended to clarify the terms used. Unless otherwise specified, a term should not be considered ambiguous. Rather, any undefined term should be interpreted in accordance with its general meaning as understood by one of ordinary skill in the art to which this invention pertains.

[0028] As used herein, the term "alkyl" refers to a straight or branched chain saturated hydrocarbon of from 1 to 10 carbon atoms, which can be optionally substituted with a number of substituents as further described herein. Examples of "alkyl" as used herein include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, isobutyl, n-butyl, sec-butyl, t-butyl, isoamyl, n-amyl, neopentyl, and n-hexyl.

[0029] The number of carbon atoms in an alkyl group is indicated by the phrase "C x-y "alkyl" means that the alkyl group as defined herein contains x to y (inclusive of x and y) carbon atoms. Similar term definitions apply to other terms and ranges. Thus, C 1-6 "alkyl" means that the alkyl group as defined herein contains x to y (inclusive of x and y) carbon atoms. Similar term definitions apply to other terms and ranges. Thus, C

[0030] As used herein, the term "alkylene" refers to a straight or branched chain divalent saturated hydrocarbon residue of from 1 to 10 carbon atoms, which can be optionally substituted with a number of substituents as further described herein. Examples of "alkylene" as used herein include, but are not limited to, methylene, ethylene, n-propylene, 1-methylethylene, 2-methylethylene, dimethylmethylene, n-butylene, 1-methyl-n-propylene, and 2-methyl-n-propylene.

[0031] The number of carbon atoms in an alkylene group is indicated by the phrase "C x-y "alkylene" means that the alkylene group as defined herein contains x to y (inclusive of x and y) carbon atoms. Similar term definitions apply to other terms and ranges. Thus, C 1-4 "alkylene" means that the alkylene group as defined herein contains x to y (inclusive of x and y) carbon atoms. Similar term definitions apply to other terms and ranges. Thus, C

[0032] As used herein, the term "cycloalkyl" refers to a saturated 3- to 10-membered cyclic hydrocarbon ring, which can be optionally substituted with a number of substituents as further described herein. The "cycloalkyl" group is a monocyclic, bicyclic, or tricyclic ring. Examples of "cycloalkyl" groups as used herein include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, and adamantyl.

[0033] The number of carbon atoms in a cycloalkyl group is indicated by the phrase "C x-y "alkylene" means that the alkylene group as defined herein contains x to y (inclusive of x and y) carbon atoms. Similar term definitions apply to other terms and ranges. Thus, C 3-10Cycloalkyl represents a cycloalkyl group as described above having 3 to 10 carbons, for example including, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, and adamantyl.

[0034] As used herein, the term "heterocycle" or "heterocyclyl" refers to an optionally substituted, monocyclic, or polycyclic, saturated ring system containing one or more heteroatoms. The "heterocycle" or "heterocyclyl" group can be optionally substituted with the number of substituents allowed as further described herein. The term "heterocycle" or "heterocyclyl", as used herein, does not include ring systems containing one or more aromatic rings. Examples of heteroatoms include nitrogen, oxygen, or sulfur atoms, including N-oxides, sulfur oxides, and sulfur dioxides. Typically, the ring is 3 to 12 membered. The ring can be optionally fused to one or more additional heterocyclic or cycloalkyl rings. Examples of "heterocycle" groups, as used herein, include, but are not limited to, tetrahydrofuran, tetrahydropyran, 1,4-dioxane, 1,3-dioxane, piperidine, pyrrolidine, morpholine, tetrahydrothiopyran, and tetrahydrothiophene, wherein attachment can occur at any point on the ring as long as the attachment is chemically feasible. Thus, for example, "morpholine" refers to morpholin-2-yl, morpholin-3-yl, and morpholin-4-yl.

[0035] As used herein, in the case of "heterocycle" or "heterocyclyl" as a possible substituent, the "heterocycle" or "heterocyclyl" group can be attached through a carbon atom or any heteroatom as long as the attachment at that point is chemically feasible. For example, "heterocyclyl" can include pyrrolidin-1-yl, pyrrolidin-2-yl, and pyrrolidin-3-yl. In the case of "heterocycle" or "heterocyclyl" groups containing a nitrogen atom in the ring, attachment through the nitrogen atom can alternatively be indicated by suffixing the ring name with "-ino". For example, pyrrolidino refers to pyrrolidin-1-yl.

[0036] As used herein, the term "halogen" or "halo" refers to fluorine, chlorine, bromine, or iodine.

[0037] As used herein, the term "oxo" refers to a >C=O substituent. In the case of an oxo-substituted cycloalkyl group (e.g., 3-oxo-cyclobutyl), the substituted group is still intended to be a saturated group.

[0038] As used herein, the term "heteroaryl" refers to a 5- to 14-membered, optionally substituted, monocyclic or polycyclic ring system, which contains at least one aromatic ring and contains one or more heteroatoms. The "heteroaryl" group can be optionally substituted with the permissible number of substituents, as further described herein. In polycyclic "heteroaryl" groups containing at least one aromatic ring and at least one non-aromatic ring, the aromatic ring(s) need not contain a heteroatom. Thus, for example, "heteroaryl" as used herein will include indolyl. In addition, the point of attachment can be attached to any ring in the ring system, regardless of whether the ring containing the point of attachment is aromatic or contains a heteroatom. Thus, for example, "heteroaryl" as used herein will include indolin-1-yl, indolin-3-yl, and indolin-5-yl. Examples of heteroatoms include nitrogen, oxygen, or sulfur atoms, including N-oxides, sulfoxides, and sulfones, where feasible. Examples of "heteroaryl" groups as used herein include, but are not limited to, furanyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 1,2,4-triazolyl, pyrazolyl, pyridyl, pyridazinyl, pyrimidinyl, indolyl, isoindolyl, benzo[b]thienyl, benzoimidazolyl, benzothiazolyl, pteridinyl, and phenoxazinyl, where attachment can occur at any point on the ring, provided that the attachment is chemically feasible. Thus, for example, "thiazolyl" refers to thiazol-2-yl, thiazol-4-yl, and thiazol-5-yl.

[0039] As used herein, where "heteroaryl" is a possible substituent, the "heteroaryl" group can be attached via a carbon atom or any heteroatom, provided that the attachment at that point is chemically feasible.

[0040] As used herein, the term "heterocyclyl" refers to an optionally substituted, saturated or unsaturated, monocyclic or polycyclic ring system, which contains one or more heteroatoms. The "heterocyclyl" group can be optionally substituted with the permissible number of substituents, as further described herein. The point of attachment can be attached to any ring in the ring system, regardless of whether the ring containing the point of attachment is saturated or unsaturated, or contains a heteroatom. Thus, for example, "heterocyclyl" as used herein will include pyrrolidinyl, piperidinyl, piperazinyl, and morpholinyl. Examples of heteroatoms include nitrogen, oxygen, or sulfur atoms, including N-oxides, sulfoxides, and sulfones, where feasible. Examples of "heterocyclyl" groups as used herein include, but are not limited to, wherein the asterisk indicates the point of attachment.

[0041] As used herein, the term "heteroaryl" refers to a 5- to 14-membered, optionally substituted, monocyclic or polycyclic ring system, which contains at least one aromatic ring and contains one or more heteroatoms. The "heteroaryl" group can be optionally substituted with the permissible number of substituents, as further described herein. In polycyclic "heteroaryl" groups containing at least one aromatic ring and at least one non-aromatic ring, the aromatic ring(s) need not contain a heteroatom. Thus, for example, "heteroaryl" as used herein will include indolyl. In addition, the point of attachment can be attached to any ring in the ring system, regardless of whether the ring containing the point of attachment is aromatic or contains a heteroatom. Thus, for example, "heteroaryl" as used herein will include indolin-1-yl, indolin-3-yl, and indolin-5-yl. Examples of heteroatoms include nitrogen, oxygen, or sulfur atoms, including N-oxides, sulfoxides, and sulfones, where feasible. Examples of "heteroaryl" groups as used herein include, but are not limited to, furanyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 1,2,4-triazolyl, pyrazolyl, pyridyl, pyridazinyl, pyrimidinyl, indolyl, isoindolyl, benzo[b]thienyl, benzoimidazolyl, benzothiazolyl, pteridinyl, and phenoxazinyl, where attachment can occur at any point on the ring, provided that the attachment is chemically feasible. Thus, for example, "thiazolyl" refers to thiazol-2-yl, thiazol-4-yl, and thiazol-5-yl. wherein the asterisk indicates the point of attachment.

[0042] Various other chemical terms or abbreviations have the standard meanings as understood by those skilled in the art. For example: "hydroxy" means -OH; "methoxy" means -OCH3; "cyano" means -CN; "amino" means -NH2; "methylamino" means -NHCH3; "sulfonyl" means -SO2-; "carbonyl" means -C(O)-; "carboxy" or "carboxyl" means -CO2H, and the like. Furthermore, where a name describes more than one moiety, the moiety described first is further from the point of attachment than any moiety described later. Thus, the term such as "methylaminocarbonylmethyl" means -CH2-C(O)-NH-CH3.

[0043] As used herein, the term "substituted" means that one or more hydrogens of the designated moiety are replaced by a named substituent or substituents, with the proviso that the substitution results in a stable or chemically feasible compound, using the allowable number of substitutions, unless otherwise indicated. A stable compound or chemically feasible compound is one whose chemical structure remains essentially unchanged under the temperature and humidity conditions likely to be encountered in normal storage and use, and in the absence of water or other chemically reactive conditions, for a period of at least one week; or maintains its integrity long enough to be used for therapeutic or prophylactic administration to a subject. As used herein, the phrase "substituted one or more times with" or "substituted one or more times by" means the number of substituents equal to one to the maximum possible number of substituents based on the number of available bonding sites, with the proviso that the stability and chemical feasibility conditions described above are met.

[0044] As used herein, the various functional groups described are to be understood as having the point of attachment at the functional group with the hyphen or dash (-) or asterisk (*). In other words, in the case of -CH2CH2CH3, the point of attachment is understood to be the CH2group on the left. If a described group does not have an asterisk or hyphen, the point of attachment is indicated by the general ordinary meaning of the group described.

[0045] Where any variable occurs more than one time in any constituent (e.g. a d ) or in multiple constituents, its definition in each occurrence is independent of its definition at every other occurrence.

[0046] As used herein, polyatomic divalent species are read from left to right. For example, if the specification or claims recite A-D-E and D is defined as -OC(O)-, the group resulting from the substitution of D is: A-OC(O)-E and not A-C(O)O-E.

[0047] As used herein, the term "optionally" means that the subsequently described event can or can not occur.

[0048] As used herein, "administering" or "administration" means introducing, such as introducing a compound or composition into a subject. The term is not limited to any particular mode of delivery, and can include, e.g., intravenous delivery, transdermal delivery, oral delivery, nasal delivery, and rectal delivery. In addition, depending on the mode of delivery, the administration can be performed by various individuals, including, e.g., a medical caregiver (e.g., a physician, a nurse, etc.), a pharmacist, or the subject (i.e., self-administration).

[0049] As used herein, "treatment" or "treating" or "treat" can refer to one or more of: delaying the progression of a disease or disorder, managing a disease or disorder, delaying the onset of a disease or disorder, ameliorating one or more symptomatic features of a disease or disorder, or delaying the recurrence of a disease or disorder or symptomatic features thereof, depending on the nature of the disease or disorder and its symptomatic features. "Treatment" or "treating" or "treat" can also refer to the inhibition of physical (e.g., stabilization of identifiable symptoms), physiological (e.g., stabilization of physical parameters), or both, and to the inhibition of at least one physical parameter that can or can not be identifiable to the subject. In certain embodiments, "treatment" or "treating" or "treat" refers to delaying the onset of a disease or at least one or more symptoms thereof in a subject who can be predisposed to the disease or who does not yet experience or exhibit symptoms of the disease.

[0050] As used herein, "subject" can refer to any mammal such as, but not limited to, a human. In one embodiment, the subject is a human. In yet another embodiment, the host is a human who exhibits one or more symptomatic features of a disease or disorder. The term "subject" does not require that it have any particular status with respect to any hospital, clinic, or research facility (e.g., as an inpatient, a research participant, etc.). In one embodiment, the subject can be a "subject in need thereof.

[0051] "Therapeutically effective amount" refers to the amount of a compound that, when administered to a subject for treating a disease or at least one clinical symptom of a disease, is sufficient to affect such treatment for the disease or its symptoms. The "therapeutically effective amount" can vary depending, for example, on the compound, the disease and / or disease symptoms, the severity of the disease, and / or the symptoms of the disease or disorder, the age, weight, and / or health of the subject to be treated, and the judgment of the prescribing physician. An appropriate amount in any given instance can be determined by one of skill in the art or can be determined by routine experimentation.

[0052] As used herein, the term "compound" of the present application includes the free acid, the free base, and any salt of the compound of Formula (I). Thus, a phrase such as "the compound of embodiment 1" or "the compound of claim 1" refers to any free acid, free base, and any salt of the compound of embodiment 1 or claim 1, respectively, that is encompassed.

[0053] II. Methods of Treatment

[0054] A. Treatment of SCD and Related Diseases with Compounds of the Invention

[0055] In one embodiment, the present application provides a method of increasing HbF expression in cells, such as red blood cells or retinal pigment epithelial (RPE) cells, by contacting the cells with a therapeutically effective amount of a compound of the present application. In other embodiments, the present application provides a method of increasing HbF expression in cells by administering a compound of the present application to a subject in need thereof. In embodiments, the increased expression of HbF is such that HbF is greater than or equal to 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 16%, 18%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, or 90% of the total hemoglobin in the subject or in a sample taken from the subject. In embodiments, the increased expression of HbF is such that HbF is increased by at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 16%, 18%, 20% percentage points of the total hemoglobin in the subject or in a sample taken from the subject relative to a baseline sample taken from the subject prior to treatment. In another embodiment, where the subject is a human less than 19 years of age, the increased expression of HbF is such that HbF is greater than or equal to 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 16%, 18%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, or 90% of the total hemoglobin in the subject or in a sample taken from the subject. The method can be used to compensate for a mutation in the human beta-globin gene, such as a mutation that results in the expression of the HbS form of hemoglobin, in cells having one or more mutations in the beta-globin gene or its expression control sequences. Compensating for the mutation includes, but is not limited to, increasing the amount of HbF and decreasing the amount of HbS in the subject compared to an untreated subject or the subject prior to treatment. In another embodiment, the method of treatment results in an increase in the ratio of HbF to HbS expressed in the cells of a subject in need thereof. The method can be used to treat sickle cell disease, such as sickle cell anemia and other hemoglobinopathies or thalassemia, and complications associated with SCD, such as retinopathy.

[0056] In another embodiment, the present application provides a method of inhibiting HbS polymerization, increasing the level of dissolved oxygen in the blood of a subject, reducing the level of reactive oxygen species (ROS), or any combination thereof by administering a compound of the present application to a subject in need thereof.

[0057] In another embodiment, the present application provides a method of reducing sickling, reducing the incidence or probability of a pain crisis, reducing the incidence or probability of a pain crisis requiring hospitalization, reducing the incidence of chest syndrome, reducing the number of transfusion events, reducing the number of units transfused per event, or any combination thereof in response to reduced air pressure, reduced atmospheric pressure, reduced oxygen partial pressure, or hypoxia by administering a compound of the present application to a subject in need thereof. The reduction in incidence or probability can be for more than one week, one month, or one year.

[0058] In another embodiment, the present application provides a method of treatment comprising administering a compound (or salt) of any one of embodiments 1 to 250 to a subject. In another embodiment, the present application provides a method of treatment comprising administering 0.1 mg to 2 grams of a compound (or salt) of any one of embodiments 1 to 250 to a subject.

[0059] In each of the methods described above or below, the compound (or salt) of any one of embodiments 1 to 250 can be administered to the subject as part of a pharmaceutical formulation, as described herein.

[0060] In each of the methods described herein, the method can further comprise the step of determining whether the subject has one or more genetic alterations associated with SCD or first determining whether the subject has a biochemical or morphological alteration associated with SCD.

[0061] In each of the methods described herein, the method can further comprise the step of determining whether the administration of a compound of the present application has increased expression of HbF, reduced a biomarker associated with SCD (e.g., ROS), or reduced a symptom associated with SCD. The method can further comprise the step of administering a higher dose of a compound of the present application if the subject does not have increased expression of HbF, does not have reduced a biomarker associated with SCD, such as ROS, or does not have reduced a symptom associated with SCD.

[0062] B. Treatment in combination with HU or Nrf2 activators

[0063] The methods described herein for treating SCD or complications thereof can further comprise administering or alternating administration of a compound of the present application in combination with HU or Nrf2 activators. The combination can be administered in an amount effective to induce or increase expression of HbF.

[0064] C. Diseases to be treated

[0065] The compounds of the present application and the combinations described herein can be used to treat a subject having one or more mutations in the beta-globin gene (HBB gene). Mutations in the beta-globin gene can result in sickle cell disease, beta-thalassemia, or a related disease or condition thereof. As discussed in more detail below, mutations in the beta-globin gene can be identified prior to or after the clinical symptoms of the disease manifest. The compositions can be administered to a subject having one or more mutations in the beta-globin gene prior to or after the clinical symptoms appear. Thus, in some embodiments, the compositions are administered to a subject who has been diagnosed with one or more mutations in the beta-globin gene but has not yet manifested clinical symptoms. In some embodiments, the compositions are administered to a subject who exhibits one or more symptoms of a disease, condition, or syndrome associated with or caused by one or more mutations in the beta-globin gene.

[0066] 1. Sickle Cell Disease

[0067] Sickle cell disease (SCD) is typically caused by a mutation in which a thymine replaces an adenine in the sixth codon of the hemoglobin beta chain gene (i.e., GAG to GTG of the HBB gene). This mutation results in a substitution of valine for glutamic acid at position 6 of the Hb beta chain. The resulting Hb, called HbS, has the physical property of forming polymers under deoxygenated conditions. SCD is typically an autosomal recessive disease. Thus, in some embodiments, the disclosed compositions and methods are used to treat a subject who is homozygous for an autosomal recessive mutation in the hemoglobin beta chain gene (i.e., a sickle hemoglobin (HbS) homozygote). Also known as HbSS disease or sickle cell anemia (the most common form), S globin homozygote subjects typically exhibit a severe or moderately severe phenotype, and have the shortest survival time of the hemoglobinopathies.

[0068] Sickle cell trait or carrier status is the heterozygous form, which is characterized by the presence of about 40% HbS, no anemia, inability to concentrate urine (isosthenuria), and hematuria. It can become a pathologic risk factor under conditions that result in hypoxia. Thus, in some embodiments, the disclosed compositions and methods are used to treat a subject who is heterozygous for an autosomal recessive mutation in the hemoglobin beta chain gene (i.e., an HbS heterozygote).

[0069] 2. Beta-Thalassemia

[0070] β-thalassemia is a group of inherited blood disorders caused by a variety of mutational mechanisms that result in reduced or absent β-globin synthesis and accumulation of unpaired, insoluble a-chain polymers, leading to ineffective erythropoiesis, accelerated red blood cell destruction, and severe anemia. Subjects with β-thalassemia exhibit a range of phenotypes from severe anemia to clinically asymptomatic individuals. The genetic mutations present in β-thalassemia are diverse and can be caused by a variety of different mutations. Mutations can involve a single base substitution or deletion or insertion within, near, or upstream of the β-globin gene. For example, mutations occur in the promoter region preceding the β-globin gene or result in the production of aberrant splice variants. Examples of thalassemia include thalassemia minor, thalassemia intermedia, and thalassemia major.

[0071] 3. Sickle cell-related diseases

[0072] Although sickle cell trait carriers do not have SCD, individuals with one copy of HbS and one copy of a gene encoding another abnormal variant of hemoglobin (such as HbC or Hbβ-thalassemia) have a less severe form of the disease. Subjects who are double heterozygotes for HbS and HbC (HbSC disease) are typically characterized by symptoms of moderate clinical severity. Another common structural variant of β-globin is hemoglobin E or hemoglobin E (HbE). As described below, subjects who are double heterozygotes for HbS and HbE have HbS / HbE syndrome, which often results in a phenotype similar to HbS / β+ thalassemia.

[0073] Certain mutations in the β-globin gene result in other structural variants of hemoglobin or result in insufficient quantities of β-globin being produced. These types of mutations are referred to as β-thalassemia mutations. Absence of β-globin is referred to as β-zero (β°) thalassemia. Subjects who are double heterozygotes for HbS and β° thalassemia (i.e., HbS / β° thalassemia) can present with symptoms that are clinically indistinguishable from sickle cell anemia. Reduced quantities of β-globin are referred to as β-plus (β+) thalassemia. Subjects who are double heterozygotes for HbS and β+ thalassemia (i.e., HbS / β+ thalassemia) can have clinical symptoms of mild to moderate severity and vary between ethnic groups. Rare combinations of HbS with other abnormal hemoglobins include HbD Los Angeles, G-Philadelphia, HbO Arab, and others.

[0074] Thus, in some embodiments, the disclosed compositions and methods are used to treat a subject having an HbS / β° genotype, an HbS / β+ genotype, an HBSC genotype, an HbS / HbE genotype, HbD Los Angeles, G-Philadelphia genotype, or an abHbO Arab genotype.

[0075] As noted above, retinopathy caused by SCD can also be treated by administering an effective amount of a compound of the application, optionally in an amount effective to induce HbF expression in retinal cells, e.g., RPE cells, in combination or alternation with HU or an Nrf2 activator. Optionally administering a compound of the application in combination with HU or an Nrf2 activator can reduce or inhibit the formation of occlusions in the peripheral retina of a sickle cell patient.

[0076] 4. Non-erythroid associated diseases

[0077] While red blood cells are the primary producers of hemoglobin, reports indicate that other non-hematopoietic cells, including but not limited to macrophages, retinal pigment cells, and alveolar epithelial cells, such as alveolar type II (ATII) cells and Clara cells, also synthesize hemoglobin. In some embodiments, the compositions disclosed herein are used to increase HbF expression in non-erythroid cells, including but not limited to macrophages, retinal pigment cells, and alveolar epithelial cells, such as alveolar type II (ATII) cells and Clara cells. In some embodiments, the compositions disclosed herein are used to increase HbF expression at interfaces where oxygen-carbon dioxide diffusion occurs, including but not limited to the eye and the lung. In some embodiments, the compositions are used in an amount effective to induce, increase, or enhance hemoglobin synthesis in retinal pigment cells to prevent, reduce, or lessen one or more symptoms of age-related macular degeneration or diabetic retinopathy.

[0078] D. Symptoms of SCD, beta-thalassemia, and related diseases

[0079] In some embodiments, the compositions disclosed herein are administered to a subject in an amount effective to treat one or more symptoms of sickle cell disease, beta-thalassemia, or a related disorder.

[0080] Beta-thalassemia can include symptoms such as anemia, fatigue and weakness, pale or yellowish skin, abdominal prominence with splenomegaly and hepatomegaly, dark urine, facial skeletal abnormalities, poor growth, and loss of appetite.

[0081] In a subject with sickle cell disease or a related disease, physiological changes in red blood cells can result in disease with the following signs: (1) hemolytic anemia; (2) vaso-occlusive crisis; (3) multi-organ damage due to microinfarcts, including the heart, bones, spleen, and central nervous system.

[0082] III. Compositions for treating SCD and related disorders

[0083] A. Compounds of the invention (compounds of Formula (I))

[0084] Compounds of Formula (I) have the structure shown below

[0085]

[0086] wherein

[0087] X 1 is =N- or =CH-;

[0088] X 2 is =C(R 1 )- and X 3 is =C(-L-G)-; or X 2 is =C(-L-G)- and X 3 is =C(R 1 )-;

[0089] G is hydrogen, -C 1-8 alkyl, -C 3-10 cycloalkyl, -C 1-6 alkylene-C 3-10 cycloalkyl, heterocyclyl, -C 1-6 alkylene-C 3-10 heterocyclyl, phenyl, heteroaryl, or NR h R k , wherein said alkyl, alkylene, cycloalkyl, heterocyclyl, phenyl, and heteroaryl are optionally substituted one or more times with substituents independently selected from R c ; or G is -CH2Y 3 , -CH2CH2Y 3 , -CH2CH2CH2Y 3 , -CH(CH3)CH2Y 3 , -CH2CH(Y 3 )CH3, -CH(Y 3 )CH3, -CH2C(Y 3 )(CH3)2, -C(Y 3 )(CH3)2, or wherein Y 3is cyclopropyl, -CF3, -OCF3, -OCH3, -OCH2CH3, -F, -Cl, -OH, -0(CH2)2-OH, -0(CH2)2-F, -SCH3, -S(O)2-CH3, -SCH2CH3, -S(O)2CH2CH3, -NH-CH3, -NH-CH2CH3, -N(CH3)2, tetrahydropyran-4-yl, tetrahydrofuran-2-yl, morpholin-2-yl, morpholin-4-yl, piperidin-1-yl, 4-hydroxy-piperidin-1-yl, 3-hydroxy-piperidin-1-yl, -NH-C(O)-CH3, -NH-C(O)-CH2CH3, tetrahydrofuran-2-yl-methyloxy, or -C(O)-Y 4 wherein Y 4 is -OH, -OCH3, -OCH2CH3, -OC(CH3)3, -NH2, -NH-CH3, -NH-CH2CH3, -N(CH3)2, -N(CH2CH3)2, morpholin-4-yl, 4-methyl-piperazin-1-yl, pyrrolidin-1-yl, or piperazin-1-yl;

[0090] L is -CH2-C(O)N(R 6 )-, -C(O)N(R 6 )-, -C(O)-O-, -SO2-, -C(O)-, heteroarylene, optionally substituted one or more times by a substituent independently selected from R x , or heterocyclylene, optionally substituted one or more times by a substituent independently selected from R x ; or the group -L-G is -cyano;

[0091] R 1 is hydrogen, R a , phenyl, or heteroaryl, wherein phenyl and heteroaryl are optionally substituted one or more times by a substituent independently selected from R x ;

[0092] R 2 is R b ;

[0093] R 3 is hydrogen, -C 1-6 alkyl, or -C 1-6 alkylene-C 3-10 cycloalkyl, wherein said alkyl, alkylene, and cycloalkyl are optionally substituted one or more times by a substituent independently selected from R z ;

[0094] R 4 is -C 1-6 alkyl or -C 1-6 alkylene-C 3-10cycloalkyl, wherein said alkyl, alkylene, and cycloalkyl are optionally substituted one or more times by substituents independently selected from R y ;

[0095] R 6 is hydrogen, -C 1-6 alkyl, -C 1-6 alkylene-C 3-10 cycloalkyl, wherein said alkyl, alkylene, and cycloalkyl are optionally substituted one or more times by substituents independently selected from R x ;

[0096] R a is

[0097] a) -halo,

[0098] b) -C 1-6 alkyl,

[0099] c) -C 3-10 cycloalkyl,

[0100] d) -heterocyclyl,

[0101] e) -cyano,

[0102] f) -CF3,

[0103] g) -OCF3,

[0104] h) -O-R d ,

[0105] i) -S(O) w -R d ,

[0106] j) -S(O)2O-R d ,

[0107] k) -NR d R e ,

[0108] l) -C(O)-R d ,

[0109] m) -C(O)-O-R d ,

[0110] n) -OC(O)-R d ,

[0111] o) -C(O)NR d R e ,

[0112] p) -C(O)-heterocyclyl,

[0113] q) -NRd C(O)R e ,

[0114] r)-OC(O)NR d R e ,

[0115] s)-NR d C(O)OR d , or

[0116] t)-NR d C(O)NR d R e ,

[0117] wherein said alkyl, cycloalkyl, and heterocyclyl are optionally substituted one or more times with substituents independently selected from R y ;

[0118] R b is

[0119] a)-halo,

[0120] b)-C 1-6 alkyl,

[0121] c)-C 3-10 cycloalkyl,

[0122] d)-heterocyclyl,

[0123] e)-phenyl,

[0124] f)-heteroaryl,

[0125] g)-cyano,

[0126] h)-CF3,

[0127] i)-OCF3,

[0128] j)-O-R f ,

[0129] k)-S(O) w -R f ,

[0130] l)-S(O)2O-R f ,

[0131] m)-NR f R g ,

[0132] n)-C(O)-R f ,

[0133] o)-C(O)-O-R f ,

[0134] p) -OC(O)-R f ,

[0135] q) -C(O)NR f R g ,

[0136] r) -C(O)-heterocyclyl,

[0137] s) -NR f C(O)R g ,

[0138] t) -OC(O)NR f R g ,

[0139] u) -NR f C(O)OR f , or

[0140] v) -NR f C(O)NR f R g ,

[0141] wherein said alkyl, cycloalkyl, heterocyclyl, phenyl, and heteroaryl are optionally substituted one or more times with a substituent selected independently from R z ;

[0142] R c is

[0143] a) -halo,

[0144] b) -C 1-6 alkyl,

[0145] c) -C 3-10 cycloalkyl,

[0146] d) -heterocyclyl,

[0147] e) -cyano,

[0148] f) -CF3,

[0149] g) -OCF3,

[0150] h) -O-R h ,

[0151] i) -S(O) w -R h ,

[0152] j) -S(O)2O-R h ,

[0153] k) -NR hR k ,

[0154] l)-C(O)-R h ,

[0155] m)-C(O)-O-R h ,

[0156] n)-OC(O)-R h ,

[0157] o)-C(O)NR h R k ,

[0158] p)-C(O)-heterocyclyl,

[0159] q)-NR h C(O)R k ,

[0160] r)-OC(O)NR h R k ,

[0161] s)-NR h C(O)OR k ,

[0162] t)-NR h C(O)NR h R k ,

[0163] u)-NR h S(O) w R k ,

[0164] v)-phenyl,

[0165] w)-heteroaryl, or

[0166] x)-O-(C 1-4 alkylene)-O-(C 1-4 alkylene)-N(R h )C(O)-OR k ,

[0167] wherein said alkylene, alkyl, cycloalkyl, heterocyclyl, phenyl, and heteroaryl are optionally substituted one or more times with a substituent independently selected from R x ;

[0168] R d and R e are independently hydrogen, C 1-6 alkyl, or C 3-10 cycloalkyl, wherein said alkyl and cycloalkyl are optionally substituted with a substituent independently selected from R ysubstituents selected from the group consisting of R d and R e together with the nitrogen atom to which they are both attached can optionally form a heterocyclic ring selected from the group consisting of azetidinyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, piperidinyl, piperazinyl, morpholino, thiomorpholino, and azepanyl, wherein each ring is optionally substituted one or more times with a substituent selected from the group consisting of R y ;

[0169] R f and R g are independently hydrogen, C 1-6 alkyl, C 3-10 cycloalkyl, phenyl, or heteroaryl, wherein the alkyl, cycloalkyl, phenyl, and heteroaryl are optionally substituted one or more times with a substituent selected from the group consisting of R z ; or, if R f and R g are both attached to the same nitrogen atom, together with the nitrogen atom to which they are both attached can optionally form a heterocyclic ring selected from the group consisting of azetidinyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, piperidinyl, piperazinyl, morpholino, thiomorpholino, and azepanyl, wherein each ring is optionally substituted one or more times with a substituent selected from the group consisting of R z ;

[0170] R h and R k are independently hydrogen, C 1-6 alkyl, C 3-10 cycloalkyl, heterocyclyl, phenyl, or heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, phenyl, and heteroaryl are optionally substituted one or more times with a substituent selected from the group consisting of R x ; or, if R h and R k are both attached to the same nitrogen atom, together with the nitrogen atom to which they are both attached can optionally form a heterocyclic ring selected from the group consisting of azetidinyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, piperidinyl, piperazinyl, morpholino, thiomorpholino, and azepanyl, wherein each ring is optionally substituted one or more times with a substituent selected from the group consisting of R x ;

[0171] R y is

[0172] a) -halogen,

[0173] b) -NH2,

[0174] c) -cyano,

[0175] d) -carboxy,

[0176] e) -hydroxy,

[0177] f) -mercapto,

[0178] g) -CF3,

[0179] h) -OCF3,

[0180] i) -C(O)-NH2,

[0181] j) -S(O)2-NH2,

[0182] k) oxo,

[0183] 1) -C 1-6 alkyl, optionally substituted one or more times with substituents independently selected from the group consisting of: halo, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0184] m) -heterocyclyl, optionally substituted one or more times with substituents independently selected from the group consisting of: halo, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0185] n) -C 3-10 cycloalkyl, optionally substituted one or more times with substituents independently selected from the group consisting of: halo, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0186] o) -O-C 1-6 alkyl, optionally substituted one or more times with substituents independently selected from the group consisting of: halo, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0187] p) -O-C 3-10 cycloalkyl, optionally substituted one or more times with substituents independently selected from the group consisting of: halo, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0188] q) -NH-C 1-6 alkyl, optionally substituted one or more times with substituents independently selected from the group consisting of halogen, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0189] r) -N(C 1-6 alkyl)2, optionally substituted one or more times with substituents independently selected from the group consisting of halogen, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0190] s) -C(O)-C 1-6 alkyl, optionally substituted one or more times with substituents independently selected from the group consisting of halogen, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0191] t) -C(O)-O-C 1-6 alkyl, optionally substituted one or more times with substituents independently selected from the group consisting of halogen, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0192] u) -S-C 1-6 alkyl, optionally substituted one or more times with substituents independently selected from the group consisting of halogen, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0193] v) -S(O)2-C 1-6 alkyl, optionally substituted one or more times with substituents independently selected from the group consisting of halogen, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0194] w) -C(O)-NH-C 1-6 alkyl, optionally substituted one or more times with substituents independently selected from the group consisting of halogen, -OH, -O-C 1-6alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0195] x)-C(O)-N(C 1-6 alkyl)2, optionally substituted one or more times with a substituent independently selected from the group consisting of: halo, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0196] y)-S(O)2-NH-C 1-6 alkyl, optionally substituted one or more times with a substituent independently selected from the group consisting of: halo, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0197] z)-S(O)2-N(C 1-6 alkyl)2, optionally substituted one or more times with a substituent independently selected from the group consisting of: halo, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0198] aa)-NH-C(O)-C 1-6 alkyl, optionally substituted one or more times with a substituent independently selected from the group consisting of: halo, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2, or

[0199] bb)-NH-S(O)2-C 1-6 alkyl, optionally substituted one or more times with a substituent independently selected from the group consisting of: halo, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2;

[0200] R x is

[0201] a)-R y

[0202] b)-phenyl, optionally substituted one or more times with a substituent independently selected from the group consisting of: halo, -OH, -O-C 1-6 alkyl, -NH2, -NH-C1-6 alkyl, and -N(C 1-6 alkyl)2,

[0203] c) -heteroaryl, optionally substituted one or more times with a substituent independently selected from the group consisting of halogen, -OH, -O-C 1-6 alkyl, -NH2, -NH-C 1-6 alkyl, and -N(C 1-6 alkyl)2,

[0204] d) -O-phenyl,

[0205] e) -O-heteroaryl,

[0206] f) -C(O)-phenyl,

[0207] g) -C(O)-heteroaryl,

[0208] h) -C(O)-O-phenyl, or

[0209] i) -C(O)-O-heteroaryl;

[0210] R z is

[0211] a) -R y

[0212] b) -phenyl,

[0213] c) -heteroaryl;

[0214] d) -O-phenyl,

[0215] e) -O-heteroaryl,

[0216] f) -C(O)-phenyl,

[0217] g) -C(O)-heteroaryl,

[0218] h) -C(O)-O-phenyl, or

[0219] i) -C(O)-O-heteroaryl;

[0220] v is an integer from 0 to 4, and

[0221] w is an integer from 0 to 2.

[0222] Embodiment 2: The compound according to Embodiment 1, wherein

[0223] G is hydrogen, -C 1-8 alkyl, -C 3-10 cycloalkyl, -C 1-6 alkylene-C 3-10 cycloalkyl, heterocyclyl, phenyl, heteroaryl, or NRh R k wherein said alkyl, alkylene, cycloalkyl, heterocyclyl, phenyl, and heteroaryl are optionally substituted one or more times with substituents independently selected from R c ; or G is -CH2Y 3 , -CH2CH2Y 3 , -CH2CH2CH2Y 3 , -CH(CH3)CH2Y 3 , -CH2CH(Y 3 )CH3, -CH(Y 3 )CH3, -CH2C(Y 3 )(CH3)2, -C(Y 3 )(CH3)2, or wherein Y 3 is -cyclopropyl, -CF3, -OCF3, -OCH3, -OCH2CH3, -F, -Cl, -OH, -O(CH2)2-OH, -O(CH2)2-F, -SCH3, -S(O)2-CH3, -SCH2CH3, -S(O)2CH2CH3, -NH-CH3, -NH-CH2CH3, -N(CH3)2, tetrahydropyran-4-yl, tetrahydrofuran-2-yl, morpholin-2-yl, morpholin-4-yl, piperidin-1-yl, 4-hydroxy-piperidin-1-yl, 3-hydroxy-piperidin-1-yl, -NH-C(O)-CH3, -NH-C(O)-CH2CH3, tetrahydrofuran-2-yl-methyloxy, or -C(O)-Y 4 wherein Y 4 is -OH, -OCH3, -OCH2CH3, -OC(CH3)3, -NH2, -NH-CH3, -NH-CH2CH3, -N(CH3)2, -N(CH2CH3)2, morpholin-4-yl, 4-methyl-piperazin-1-yl, pyrrolidin-1-yl, or piperazin-1-yl;

[0224] R c is

[0225] a) halo,

[0226] b) -C 1-6 alkyl,

[0227] c) -C 3-10 cycloalkyl,

[0228] d) -heterocyclyl,

[0229] e) -cyano,

[0230] f) -CF3,

[0231] g) -OCF3,

[0232] h) -O-R h ,

[0233] i) -S(O) w -R h ,

[0234] j) -S(O)2O-R h ,

[0235] k) -NR h R k ,

[0236] l) -C(O)-R h ,

[0237] m) -C(O)-O-R h ,

[0238] n) -OC(O)-R h ,

[0239] o) -C(O)NR h R k ,

[0240] p) -C(O)-heterocyclyl,

[0241] q) -NR h C(O)R k ,

[0242] r) -OC(O)NR h R k ,

[0243] s) -NR h C(O)OR k ,

[0244] t) -NR h C(O)NR h R k ,

[0245] u) -phenyl,

[0246] v) -heteroaryl, or

[0247] w) -O-(C 1-4 alkylene)-O-(C 1-4 alkylene)-N(R h )C(O)-OR k ,

[0248] wherein said alkylene, alkyl, cycloalkyl, heterocyclyl, phenyl, and heteroaryl are optionally substituted one or more times with substituents independently selected from R x ;

[0249] R h k independently hydrogen, C 1-6 alkyl, C 3-10 cycloalkyl, phenyl, or heteroaryl, wherein said alkyl, cycloalkyl, phenyl, and heteroaryl are optionally substituted one or more times with a substituent independently selected from R x ; or, if R h and R k are both attached to the same nitrogen atom, together with the nitrogen atom to which they are attached can optionally form a heterocycloalkyl ring selected from azetidinyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, piperidinyl, piperazinyl, morpholino, thiomorpholino, and azepanyl, wherein each ring is optionally substituted one or more times with a substituent independently selected from R x ; and

[0250] R y

[0251] a) -halo,

[0252] b) -NH2,

[0253] c) -cyano,

[0254] d) -carboxy,

[0255] e) -C 1-6 alkyl, optionally substituted one or more times with halo,

[0256] f) -heterocycloalkyl, optionally substituted one or more times with halo,

[0257] g) -C 3-10 cycloalkyl, optionally substituted one or more times with halo,

[0258] h) -O-C 1-6 alkyl, optionally substituted one or more times with halo,

[0259] i) -O-C 3-10 cycloalkyl, optionally substituted one or more times with halo,

[0260] j) -hydroxy,

[0261] k) -mercapto,

[0262] l) -CF3,

[0263] m) -OCF3,

[0264] n) -C(O)-C 1-6 alkyl, optionally substituted one or more times with halo, ​​

[0265] o) -C(O) -O-C 1-6 alkyl, optionally substituted one or more times with halo,

[0266] p) -S-C 1-6 alkyl, optionally substituted one or more times with halo, or

[0267] q) -S(O)2-C 1-6 alkyl, optionally substituted one or more times with halo.

[0268] Embodiment 3: The compound according to Embodiment 2, wherein

[0269] R 3 is hydrogen.

[0270] Embodiment 4: The compound according to Embodiment 2, wherein

[0271] R 3 is methyl.

[0272] Embodiment 5: The compound according to any one of Embodiments 2 to 4, wherein

[0273] X 1 is =N-.

[0274] Embodiment 6: The compound according to any one of Embodiments 2 to 4, wherein

[0275] X 1 is =CH-.

[0276] Embodiment 7: The compound according to any one of Embodiments 2 to 6, wherein

[0277] v is an integer from 0 to 2.

[0278] Embodiment 8: The compound according to any one of Embodiments 2 to 6, wherein

[0279] v is 0 or 1.

[0280] Embodiment 9: The compound according to any one of Embodiments 2 to 6, wherein

[0281] v is 1.

[0282] Embodiment 10: The compound according to any one of Embodiments 2 to 6, wherein

[0283] v is 1 and R 2 is attached to the 5- or 6-position of the benzothiazole ring.

[0284] Embodiment 11: The compound according to any one of Embodiments 2 to 6, wherein

[0285] v is 1 and R2 It is attached to the 6-position of the benzothiazole ring.

[0286] Implementation Method 12: According to any one of Implementation Methods 2 to 6, wherein

[0287] v is 2, and 1 R 2 It is attached to the 6-position of the benzothiazole ring.

[0288] Embodiment 13: According to any one of Embodiments 2 to 6, wherein

[0289] v is 2, and R 2 It is attached to the 5- or 6-position of the benzothiazole ring.

[0290] Embodiment 14: According to any one of Embodiments 2 to 13, wherein

[0291] R 2 It is -halogen, -C 1-6 Alkyl, -CF3, -OCF3, -OR f , or -S(O) w -R f The alkyl group thereon is optionally selected from R z The substituents are substituted once or multiple times.

[0292] Embodiment 15: According to any one of Embodiments 2 to 13, wherein

[0293] R 2 It is -halogen, -methyl, -CF3, -OCF3, -SCF3, -O-heteroaryl, or -S(O)2-CH3.

[0294] Embodiment 16: According to any one of Embodiments 2 to 13, wherein

[0295] R 2 Selected from -Cl, -F, -CF3, and -OCF3.

[0296] Embodiment 17: According to any one of Embodiments 2 to 13, wherein

[0297] R 2 It is -OCF3.

[0298] Embodiment 18: According to any one of Embodiments 2 to 13, wherein

[0299] R 2 It is -CF3.

[0300] Embodiment 19: According to any one of Embodiments 2 to 13, wherein

[0301] R2 is -F.

[0302] Embodiment 20: The compound according to any one of embodiments 2 to 13, wherein

[0303] R 2 is -Cl.

[0304] Embodiment 21 : The compound according to any one of embodiments 2 to 20, wherein

[0305] R 4 is -methyl, -ethyl, -n-propyl, -i-propyl, -n-butyl, -sec-butyl, -i-butyl, -tert-butyl, -(CH2) 1-2 -OCH3, -(CH2) 1-2 -F, -(CH2) 1-2 -Cl, -(CH2) 1-2 -OCF3, -(CH2) 1-2 -NH2, -(CH2) 1-2 -CN, -(CH2) 1-2 -OH, -(CH2) 1-2 -CF3, -(CH2) 1-2 -CO2H, -(CH2) 1-2 -SH, -(CH2)1-2-SCH3, -(CH2) 1-2 -S(O)2CH3, -(CH2) 1-2 -OCH2CH3, -(CH2) 1-2 -SCH2CH3, -(CH2) 1-2 -S(O)2CH2CH3, -(CH2) 1-2 -NH-CH3, or -(CH2) 1-2 -N(CH3)2.

[0306] Embodiment 22: The compound according to any one of embodiments 2 to 21, wherein

[0307] R 4 is -methyl, -ethyl, -i-propyl, -i-butyl, -CH2CH2-OCH3, -CH2CH2-F, or -CH2CH2-NH2.

[0308] Embodiment 23: The compound according to any one of embodiments 2 to 22, wherein

[0309] R 4 is -methyl, -ethyl, -i-propyl, or -i-butyl.

[0310] Embodiment 24: The compound according to any one of embodiments 2 to 23, wherein

[0311] R 4 is -methyl.

[0312] Embodiment 25: The compound according to any one of embodiments 2 to 23, wherein

[0313] R 4 is -ethyl.

[0314] Embodiment 26: The compound according to any one of embodiments 2 to 21, wherein

[0315] R 4 is -(CH2)2-OCH3, -(CH2)2-F, -(CH2)2-Cl, -(CH2)2-OCF3, -(CH2)2-NH2, -(CH2)2-CN, -(CH2)2-OH, -(CH2)2-CF3, -(CH2)2-CO2H, -(CH2)2-SH, -(CH2)2-SCH3, or -(CH2)2-S(O)2CH3.

[0316] Embodiment 27: The compound according to any one of embodiments 2 to 26, wherein

[0317] R 1 is selected from hydrogen, -OCH3, -F, -Cl, -NH2, -cyano, -OH, -CF3, -OCF3, -SH, -S-C 1-6 alkyl, -S(O)2-C 1-6 alkyl, -CO2H, -NH-C 1-6 alkyl, -N(C 1-6 alkyl)2, and -NH-C 1-6 alkyl.

[0318] Embodiment 28: The compound according to any one of embodiments 2 to 26, wherein

[0319] R 1 is selected from -OCH3, -F, -CF3, -OCF3, -N(CH3)2, -N(CH2CH3)2, and -N(CH3)(CH2CH3).

[0320] Embodiment 29: The compound according to any one of embodiments 2 to 26, wherein

[0321] R 1 is selected from hydrogen, -OCH3, and -F.

[0322] Embodiment 30: The compound according to any one of embodiments 2 to 26, wherein

[0323] R 1 is hydrogen.

[0324] Embodiment 31 : The compound according to any one of embodiments 2 to 30, wherein

[0325] G is hydrogen, -C 1-8 alkyl, -C 3-10 cycloalkyl, -C 1-6 alkylene-C 3-8 cycloalkyl, heterocyclyl, or NR h R k , wherein said alkyl, alkylene, cycloalkyl, and heterocyclyl are optionally substituted one or more times with substituents independently selected from R c ; or G is -CH2Y 3 , -CH2CH2Y 3 , -CH2CH2CH2Y 3 , -CH(CH3)CH2Y 3 , -CH2CH(Y 3 )CH3, -CH(Y 3 )CH3, -CH2C(Y 3 )(CH3)2, -C(Y 3 )(CH3)2, or wherein Y 3 is -cyclopropyl, -CF3, -OCF3, -OCH3, -OCH2CH3, -F, -Cl, -OH, -O(CH2)2-OH, -O(CH2)2-F, -SCH3, -S(O)2-CH3, -SCH2CH3, -S(O)2CH2CH3, -NH-CH3, -NH-CH2CH3, -N(CH3)2, tetrahydropyran-4-yl, tetrahydrofuran-2-yl, morpholin-2-yl, morpholin-4-yl, piperidin-1-yl, 4-hydroxy-piperidin-1-yl, 3-hydroxy-piperidin-1-yl, -NH-C(O)-CH3, -NH-C(O)-CH2CH3, tetrahydrofuran-2-yl-methyloxy, or -C(O)-Y 4 , wherein Y 4 is -OH, -OCH3, -OCH2CH3, -OC(CH3)3, -NH2, -NH-CH3, -NH-CH2CH3, -N(CH3)2, -N(CH2CH3)2, morpholin-4-yl, 4-methyl-piperazin-1-yl, pyrrolidin-1-yl, or piperazin-1-yl;

[0326] L is -CH2-C(O)N(R 6 )-, -C(O)N(R 6 )-, -C(O)-O-, -SO2-, -C(O)-, or heterocyclylene, optionally substituted one or more times with substituents independently selected from R x ; or the group -L-G is -cyano;

[0327] R 1 is hydrogen or R a ;

[0328] R c is

[0329] a) -halogen,

[0330] b) -C 1-6 alkyl,

[0331] c) -C 3-10 cycloalkyl,

[0332] d) -heterocyclyl,

[0333] e) -cyano,

[0334] f) -CF3,

[0335] g) -OCF3,

[0336] h) -O-R h ,

[0337] i) -S(O) w -R h ,

[0338] j) -S(O)2O-R h ,

[0339] k) -NR h R k ,

[0340] l) -C(O)-R h ,

[0341] m) -C(O)-O-R h ,

[0342] n) -OC(O)-R h ,

[0343] o) -C(O)NR h R k ,

[0344] p) -C(O)-heterocyclyl,

[0345] q) -NR h C(O)R k ,

[0346] r) -OC(O)NR h R k ,

[0347] s) -NR h C(O)ORk ,

[0348] t) -NR h C(O)NR h R k , or

[0349] u) -O- (C 1-4 alkylene) -O- (C 1-4 alkylene) -N(R h )C(O) -OR k ,

[0350] wherein said alkylene, alkyl, cycloalkyl, and heterocyclyl are optionally substituted one or more times by substituents independently selected from R x ;

[0351] R h and R k are independently hydrogen, C 1-6 alkyl, or C 3-10 cycloalkyl, wherein said alkyl and cycloalkyl are optionally substituted one or more times by substituents independently selected from R x ; or, if R h and R k are both attached to the same nitrogen atom, together with the nitrogen atom to which they are attached can optionally form a heterocyclic ring selected from azetidinyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, piperidinyl, piperazinyl, morpholino, thiomorpholino, and azepanyl, wherein each ring is optionally substituted one or more times by substituents independently selected from R x ; and

[0352] R x is R y .

[0353] Embodiment 32: The compound according to any one of embodiments 2 to 31, wherein

[0354] -L-G is not -cyano.

[0355] Embodiment 33: The compound according to any one of embodiments 2 to 32, wherein

[0356] -L-G is -C(O)NR h R k .

[0357] Embodiment 34: The compound according to any one of embodiments 2 to 32, wherein

[0358] L is -C(O)N(R 6 )- or -C(O)-O-.

[0359] Embodiment 35: The compound according to any one of embodiments 2 to 32, wherein

[0360] L is -C(O)N(R 6 )-.

[0361] Embodiment 36: The compound according to any one of embodiments 2 to 32, wherein

[0362] L is not -CH2-C(O)N(R 6 )-.

[0363] Embodiment 37: The compound according to any one of embodiments 2 to 32, wherein

[0364] L is -C(O)-O-.

[0365] Embodiment 38: The compound according to any one of embodiments 2 to 32, wherein

[0366] L is -C(O)-.

[0367] Embodiment 39: The compound according to any one of embodiments 2 to 32, wherein

[0368] L is -S(O)2-.

[0369] Embodiment 40: The compound according to any one of embodiments 2 to 30, wherein

[0370] L is heteroarylene, optionally substituted one or more times by a substituent independently selected from R x .

[0371] Embodiment 41: The compound according to any one of embodiments 2 to 40, wherein

[0372] R 6 is hydrogen.

[0373] Embodiment 42: The compound according to any one of embodiments 2 to 40, wherein

[0374] R 6 is hydrogen or -methyl.

[0375] Embodiment 43: The compound according to any one of embodiments 2 to 42, wherein

[0376] G is hydrogen, -C 1-8 alkyl, -C 3-10 cycloalkyl, or -C 1-6 alkylene-C 3-8 cycloalkyl, wherein said alkyl, cycloalkyl, and alkylene are optionally substituted one or more times by a substituent independently selected from R x .

[0377] Embodiment 44: The compound according to any one of embodiments 2 to 42, wherein

[0378] G is -H, -methyl, -ethyl, -n-propyl, -isopropyl, -isobutyl, -CH2Y 3 , -CH2CH2Y 3 , -CH2CH2CH2Y 3 , -CH(CH3)CH2Y 3 , -CH2CH(Y 3 )CH3, -CH(Y 3 )CH3, -CH2C(Y 3 )(CH3)2, or -C(Y 3 )(CH3)2, wherein Y 3 is -cyclopropyl, -CF3, -OCF3, -OCH3, -OCH2CH3, -F, -OH, -O(CH2)2-OH, -O(CH2)2-F, -SCH3, -S(O)2-CH3, -SCH2CH3, -S(O)2CH2CH3, -NH-CH3, -NH-CH2CH3, -N(CH3)2, -NH-C(O)-CH3, -NH-C(O)-CH2CH3, or C(O)-Y 4 , wherein Y 4 is -OH, -OCH3, -OCH2CH3, -OC(CH3)3, -NH2, -NH-CH3, -NH-CH2CH3, -N(CH3)2, or -N(CH2CH3)2.

[0379] Embodiment 45: The compound according to any one of embodiments 2 to 42, wherein

[0380] G is -methyl, -ethyl, -n-propyl, -isopropyl, or -isobutyl, each of which is optionally substituted one or more times with a substituent independently selected from the group consisting of -CF3, -OCF3, -OCH3, -OCH2CH3, -F, -OH, -O(CH2)2-OH, -O(CH2)2-F, -SCH3, -SCH2CH3, -NH-CH3, -NH-CH2CH3, and -N(CH3)2.

[0381] Embodiment 46: The compound according to any one of embodiments 2 to 42, wherein

[0382] G is H.

[0383] Embodiment 47: The compound according to any one of embodiments 2 to 42, wherein

[0384] G is C 1-8 alkyl, optionally substituted one or more times with halogen.

[0385] Embodiment 48: The compound according to any one of embodiments 2 to 42, wherein

[0386] G is C 3-10 cycloalkyl, optionally substituted one or more times with halo.

[0387] Embodiment 49: The compound according to any one of embodiments 2 to 42, wherein

[0388] G is heterocyclyl, optionally substituted one or more times with halo.

[0389] Embodiment 50: The compound according to any one of embodiments 2 to 42, wherein

[0390] G is -C 1-6 alkylene-C 3-10 cycloalkyl, optionally substituted one or more times with halo.

[0391] Embodiment 51 : The compound according to any one of embodiments 2 to 42, wherein

[0392] G is NR h R k .

[0393] Embodiment 52: The compound according to any one of embodiments 2 to 42, wherein

[0394] G is -CH2-R c .

[0395] Embodiment 53: The compound according to any one of embodiments 2 to 42, wherein

[0396] G is -CH2CH2-R c .

[0397] Embodiment 54: The compound according to any one of embodiments 2 to 42, wherein

[0398] G is -CH2CH2CH2-R c .

[0399] Embodiment 55: The compound according to any one of embodiments 2 to 42, wherein

[0400] G is -CH(CH3)CH2-R c .

[0401] Embodiment 56: The compound according to any one of embodiments 2 to 42, wherein

[0402] G is -CH2CH(R c )CH3.

[0403] Embodiment 57: The compound according to any one of embodiments 2 to 42, wherein

[0404] G is -CH(R c )CH3.

[0405] Embodiment 58: The compound according to any one of embodiments 2 to 42, wherein

[0406] G is -CH2C(R c )(CH3)2.

[0407] Embodiment 59: The compound according to any one of embodiments 2 to 42, wherein

[0408] G is -C(R c )(CH3)2.

[0409] Embodiment 60: The compound according to any one of embodiments 2 to 42, wherein

[0410] G is imidazol-2-yl, thiazol-2yl, oxazol-2-yl, pyrazol 1-yl, furan-2-yl, thiophen-2-yl, pyrrol-1-yl, 1H-1,2,4-triazolyl-3-yl, 5-methyl-1H-1,2,4-triazolyl-3-yl, -(CH2) 1-3 -(imidazol-2-yl), -(CH2) 1-3 -(thiazol-2yl), -(CH2) 1-3 -(oxazol-2-yl), -(CH2) 1-3 -(pyrazol 1-yl), -(CH2) 1-3 -(furan-2-yl), -(CH2) 1-3 -(thiophen-2-yl), -(CH2) 1-3 -(pyrrol-1-yl), -(CH2) 1-3 -(1H-1,2,4-triazolyl-3-yl), or -(CH2) 1-3 -(5-methyl-1H-1,2,4-triazolyl-3-yl).

[0411] Embodiment 61 : The compound according to any one of embodiments 2 to 60, wherein

[0412] the compound is in its free (unsalted) form.

[0413] Embodiment 62: The compound according to any one of embodiments 2 to 60, wherein

[0414] the compound is in the form of a pharmaceutically acceptable salt.

[0415] Embodiment 63: The compound according to any one of embodiments 1 to 62, wherein

[0416] Any heterocyclic group present in the compound is selected from: azirbutan-1-yl, azirbutan-2-yl, azirbutan-3-yl, pyrrolidine-1-yl, pyrrolidine-2-yl, pyrrolidine-3-yl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothiophene-2-yl, tetrahydrothiophene-3-yl, pyrazolidine-1-yl, pyrazolidine-3-yl, pyrazolidine-4-yl, imidazolidin-1-yl, imidazolidin-2-yl, imidazolidin-4-yl, oxazolidine-2-yl, oxazolidine-3-yl, oxazolidine-4-yl, oxazolidine-5-yl, isoxazolidine- 2-yl, isoxazolidine-3-yl, isoxazolidine-4-yl, isoxazolidine-5-yl, thiazolidin-2-yl, thiazolidin-3-yl, thiazolidin-4-yl, thiazolidin-5-yl, isothiazolidine-2-yl, isothiazolidine-3-yl, isothiazolidine-4-yl, isothiazolidine-5-yl, 1,3-dioxacyclopentan-2-yl, 1,3-dioxacyclopentan-4-yl, 1,3-oxothiohexacyclopentan-2-yl, 1,3-oxothiohexacyclopentan-4-yl, 1,3-oxothiohexacyclopentan-5-yl, 1,2-dithiohexacyclopentan-3-yl, 1,2-dithiohexacyclopentan-3-yl, 1,2-dithiohexacyclopentan-3-yl pentyl-4-yl, 1,3-dithiacyclopentyl-2-yl, 1,3-dithiacyclopentyl-4-yl, piperidin-1-yl, piperidin-2-yl, piperidin-3-yl, piperidin-4-yl, tetrahydropyran-2-yl, tetrahydropyran-3-yl, tetrahydropyran-4-yl, thiocyclohexane-2-yl, thiocyclohexane-3-yl, thiocyclohexane-4-yl, piperazine-1-yl, piperazine-2-yl, morpholin-2-yl, morpholin-3-yl, morpholin-4-yl, thiomorpholin-2-yl, thiomorpholin-3-yl, thiomorpholin-4-yl, 1,4-dioxane-2-yl, 1,3- Dioxane-2-yl, 1,3-dioxane-4-yl, 1,3-dioxane-5-yl, 1,4-dithiapan-2-yl, 1,3-dithiapan-2-yl, 1,3-dithiapan-4-yl, 1,3-dithiapan-5-yl, 1,2-dithiapan-3-yl, 1,2-dithiapan-4-yl, azircycloheptane-1-yl, azircycloheptane-2-yl, azircycloheptane-3-yl, and azircycloheptane-4-yl, wherein each of these named rings may optionally be substituted once or more independently with substituents selected independently from the following: halogen, -NH2, cyano, carboxyl, C 1-4 Alkyl, C 3-10 Cycloalkyl, hydroxyl, mercapto, -CF3, -OCF3, -OC 1-4 Alkyl, -NH-C 1-4 Alkyl, -N(C) 1-4 Alkyl)2,-SC 1-4 Alkyl group, -S(O)2-C 1-4 Alkyl group, -C(O)-C 1-4 Alkyl group, -C(O)OC1-4 alkyl, -C(O)NH2, -C(O)NH-C 1-4 alkyl, and -C(O)N(C 1-4 alkyl)2, and wherein any nitrogen atom of any of these named rings can optionally be oxidized, where chemically possible, and wherein any sulfur atom of any of these named rings can optionally be oxidized one or two times, where chemically possible.

[0417] Embodiment 64: The compound according to any one of embodiments 1 to 63, wherein

[0418] Any "heteroaryl" group present in the compounds is selected from the group consisting of: 1H-pyrrol-1-yl, 1H-pyrrol-2-yl, 1H-pyrrol-3-yl, furan-2-yl, furan-3-yl, thiophen-2-yl, thiophen-3-yl, 1H-imidazol-1-yl, 1H-imidazol-2-yl, 1H-imidazol-4-yl, 1H-imidazol-5-yl, 1H-pyrazol-1-yl, 1H-pyrazol-3-yl, 1H-pyrazol-4-yl, 1H-pyrazol-5-yl, oxazol-2-yl, oxazol-4-yl, oxazol-5-yl, thiazol-2-yl, thiazol-4-yl, thiazol-5-yl, isoxazol-3-yl, isoxazol-4-yl, isoxazol-5-yl, isothiazol-3-yl, isothiazol-4-yl, isothiazol-5-yl, 1H-1,2,3-triazol-1-yl, 1H-1,2,3-triazol-4-yl, 1H-1,2,3-triazol-5-yl, 1H-1,2,4-triazol-1-yl, 1H-1,2,4-triazol-3-yl, 1H-1,2,4-triazol-5-yl, furazan-3-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, pyrazin-2-yl, 1,3,5-triazin-2-yl, 1H-indol-1-yl, 1H-indol-2-yl, 1H-indol-3-yl, 2H-isoindol-1-yl, 2H-isoindol-2-yl, quinolin-2-yl, quinolin-3-yl, quinolin-4-yl, isoquinolin-1-yl, isoquinolin-3-yl, isoquinolin-4-yl, benzoxazol-2-yl, benzothiazol-2-yl, 1H-benzoimidazol-1-yl, 1H-benzoimidazol-2-yl, benzofuran-2-yl, benzofuran-3-yl, benzothiophen-2-yl, and benzothiophen-3-yl, wherein each of these named rings can optionally be substituted one or more times with substituents independently selected from the group consisting of: halogen, -NH2, cyano, carboxyl, C 1-4 alkyl, C 3-10 cycloalkyl, hydroxy, mercapto, -CF3, -OCF3, -O-C 1-4 alkyl, -NH-C 1-4 alkyl, -N(C 1-4 alkyl)2, -S-C1-4 Alkyl group, -S(O)2-C 1-4 Alkyl group, -C(O)-C 1-4 Alkyl, -C(O)OC 1-4 Alkyl group, -C(O)NH2, -C(O)NH-C 1-4 Alkyl group, -C(O)N(C) 1-4 Alkyl)2, and phenyl.

[0419] Embodiment 65: According to any one of Embodiments 1 to 64, wherein

[0420] The arbitrary "heteroaryl" group present in the compound is selected from: 1H-pyrrole-2,5-diyl, furan-2,5-diyl, thiophene-2,5-diyl, 1H-imidazol-2,4-diyl, 1H-imidazol-2,5-diyl, oxazol-2,4-diyl, oxazol-2,5-diyl, thiazole-2,4-diyl, thiazole-2,5-diyl, 1H-1,2,4-triazole-3,5-diyl, and 2H-isoindole-1,3-diyl, wherein each of these named rings may optionally be substituted once or multiple times with substituents independently selected from the following: halogen, -NH2, cyano, carboxyl, -C 1-4 Alkyl, -C 3-10 Cycloalkyl, hydroxyl, mercapto, -CF3, -OCF3, -OC 1-4 Alkyl, -NH-C 1-4 Alkyl, -N(C) 1-4 Alkyl)2,-SC 1-4 Alkyl group, -S(O)2-C 1-4 Alkyl group, -C(O)-C 1-4 Alkyl group, -C(O)OC 1-4 Alkyl group, -C(O)NH2, -C(O)NH-C 1-4 Alkyl group, -C(O)N(C) 1-4 Alkyl)2, and phenyl.

[0421] Implementation method 66: The compound according to implementation method 1.

[0422] Embodiment 67: The compound according to Embodiment 66, wherein

[0423] R 3 It is hydrogen.

[0424] Embodiment 68: The compound according to Embodiment 66, wherein

[0425] R 3 It is a methyl group.

[0426] Embodiment 69: The compound according to Embodiment 66, wherein

[0427] R 3 is ethyl.

[0428] Embodiment 70: The compound according to Embodiment 66, wherein

[0429] R 3 is isopropyl.

[0430] Embodiment 71 : The compound according to any one of Embodiments 66 to 70, wherein

[0431] X 1 is =N-.

[0432] Embodiment 72: The compound according to any one of Embodiments 66 to 70, wherein

[0433] X 1 is =CH-.

[0434] Embodiment 73: The compound according to any one of Embodiments 66 to 72, wherein

[0435] v is 0, 1 or 2.

[0436] Embodiment 74: The compound according to any one of Embodiments 66 to 72, wherein

[0437] v is 1 or 2.

[0438] Embodiment 75: The compound according to any one of Embodiments 66 to 72, wherein

[0439] v is 1.

[0440] Embodiment 76: The compound according to any one of Embodiments 66 to 72, wherein

[0441] v is 1 and R 2 is attached to the 5-position or the 6-position of the benzothiazole ring.

[0442] Embodiment 77: The compound according to any one of Embodiments 66 to 72, wherein

[0443] v is 1 and R 2 is attached to the 6-position of the benzothiazole ring.

[0444] Embodiment 78: The compound according to any one of Embodiments 66 to 72, wherein

[0445] v is 2 and one R 2 is attached to the 6-position of the benzothiazole ring.

[0446] Embodiment 79: The compound according to any one of Embodiments 66 to 72, wherein

[0447] v is 2 and R 2 to the 5- and 6-positions of the benzothiazole ring.

[0448] Embodiment 80: The compound according to any one of embodiments 66 to 79, wherein

[0449] R 2 is -halo, -C 1-6 alkyl, -CF3, -OCF3, -O-R f , or -S(O) w -R f , wherein said alkyl is optionally substituted one or more times with substituents independently selected from R z .

[0450] Embodiment 81 : The compound according to any one of embodiments 66 to 79, wherein

[0451] R 2 is -halo, -methyl, ethyl, isopropyl, -OCH3, -OCH2CH3, -OCH(CH3)2, -CF3, -OCF3, -SCF3, -S(O)2-CH3, -O-phenyl, -O-(2-pyridyl), -O-(3-pyridyl), or -O-(4-pyridyl).

[0452] Embodiment 82: The compound according to any one of embodiments 66 to 79, wherein

[0453] R 2 is -halo, -methyl, ethyl, isopropyl, -OCH3, -OCH2CH3, -OCH(CH3)2, -CF3, -OCF3, -SCF3, -S(O)2-CH3, or -O-(3-pyridyl).

[0454] Embodiment 83: The compound according to any one of embodiments 66 to 79, wherein

[0455] R 2 is -Cl, -F, -CF3, or -OCF3.

[0456] Embodiment 84: The compound according to any one of embodiments 66 to 79, wherein

[0457] R 2 is -OCF3.

[0458] Embodiment 85: The compound according to any one of embodiments 66 to 79, wherein

[0459] R 2 is -CF3.

[0460] Embodiment 86: The compound according to any one of embodiments 66 to 79, wherein

[0461] R 2 is -F.

[0462] Embodiment 87: The compound according to any one of embodiments 66 to 79, wherein

[0463] R 2 is -Cl.

[0464] Embodiment 88: The compound according to any one of embodiments 66 to 79, wherein

[0465] R 2 is -SO2CH3.

[0466] Embodiment 89: The compound according to any one of embodiments 66 to 79, wherein

[0467] R 2 is methyl, ethyl, or isopropyl.

[0468] Embodiment 90: The compound according to any one of embodiments 66 to 79, wherein

[0469] R 2 is methyl.

[0470] Embodiment 91 : The compound according to any one of embodiments 66 to 79, wherein

[0471] R 2 is -OCH2CH3.

[0472] Embodiment 92: The compound according to any one of embodiments 66 to 79, wherein

[0473] R 2 is -O-phenyl.

[0474] Embodiment 93: The compound according to any one of embodiments 66 to 79, wherein

[0475] R 2 is -O-(2-pyridyl), -O-(3-pyridyl), or -O-(4-pyridyl).

[0476] Embodiment 94: The compound according to any one of embodiments 66 to 79, wherein

[0477] R 2 is -O-(3-pyridyl).

[0478] Embodiment 95: The compound according to any one of embodiments 66 to 94, wherein

[0479] R4 It is -methyl, -ethyl, -n-propyl, -isopropyl, -n-butyl, -sec-butyl, -isobutyl, -tert-butyl, -(CH2) 1-2 -OCH3, -(CH2) 1-2 -F, -(CH2) 1-2 -Cl, -(CH2) 1-2 -OCF3, -(CH2) 1-2 -NH2, -(CH2) 1-2 -CN, -(CH2) 1-2 -OH, -(CH2) 1-2 -CF3, -(CH2) 1-2 -CO2H, -(CH2) 1-2 -SH, -(CH2) 1-2 -SCH3, -(CH2) 1-2 -S(O)2CH3, -(CH2) 1-2 -OCH2CH3, -(CH2) 1-2 -SCH2CH3, -(CH2) 1-2 -S(O)2CH2CH3, -(CH2) 1-2 -NH-CH3, or -(CH2) 1-2 -N(CH3)2.

[0480] Embodiment 96: According to any one of Embodiments 66 to 94, wherein

[0481] R 4 It is -methyl, -ethyl, -isopropyl, -isobutyl, -CH2CH2-OCH3, -CH2CH2-F, -CH2CH2-NH2, or -CH2CH2-NH-CH3.

[0482] Embodiment 97: According to any one of Embodiments 66 to 94, wherein

[0483] R 4 It is -methyl, -ethyl, -isopropyl, or -isobutyl.

[0484] Embodiment 98: According to any one of Embodiments 66 to 94, wherein

[0485] R 4 It is a methyl group.

[0486] Embodiment 99: According to any one of Embodiments 66 to 94, wherein

[0487] R 4 It is -ethyl.

[0488] Embodiment 100: The compound according to any one of embodiments 66 to 94, wherein

[0489] R 4 is -isopropyl.

[0490] Embodiment 101 : The compound according to any one of embodiments 66 to 94, wherein

[0491] R 4 is -isobutyl.

[0492] Embodiment 102: The compound according to any one of embodiments 66 to 94, wherein

[0493] R 4 is -CH2CH2-OCH3.

[0494] Embodiment 103: The compound according to any one of embodiments 66 to 94, wherein

[0495] R 4 is -CH2CH2-F.

[0496] Embodiment 104: The compound according to any one of embodiments 66 to 94, wherein

[0497] R 4 is -CH2CH2-NH2.

[0498] Embodiment 105: The compound according to any one of embodiments 66 to 94, wherein

[0499] R 4 is -CH2CH2-NH-CH3.

[0500] Embodiment 106: The compound according to any one of embodiments 66 to 105, wherein

[0501] R 1 is independently hydrogen, -OCH3, -F, -Cl, -NH2, -cyano, -OH, -CF3, -OCF3, -SH, -S-C 1-6 alkyl, -S(O)2-C 1-6 alkyl, -CO2H, -NH-C 1-6 alkyl, -N(C 1-6 alkyl)2, or -NH-C 1-6 alkyl.

[0502] Embodiment 107: The compound according to any one of embodiments 66 to 105, wherein

[0503] R 1independently -OCH3, -F, -CF3, -OCF3, -N(CH3)2, -N(CH2CH3)2, or -N(CH3)(CH2CH3).

[0504] Embodiment 108: The compound according to any one of embodiments 66 to 105, wherein

[0505] R 1 is hydrogen, -OCH3, or -F.

[0506] Embodiment 109: The compound according to any one of embodiments 66 to 105, wherein

[0507] R 1 is hydrogen.

[0508] Embodiment 110: The compound according to any one of embodiments 66 to 105, wherein

[0509] R 1 is -F.

[0510] Embodiment 111: The compound according to any one of embodiments 66 to 105, wherein

[0511] R 1 is -OCH3.

[0512] Embodiment 112: The compound according to any one of embodiments 66 to 105, wherein

[0513] R 1 is -N(CH2CH3)2.

[0514] Embodiment 113: The compound according to any one of embodiments 66 to 112, wherein

[0515] G is hydrogen, -C 1-8 alkyl, -C 3-10 ycloalkyl, -C 1-6 alkylene-C 3-10 ycloalkyl, heterocyclyl, -C 1-6 alkylene-C 3-10 ycloalkyl, or NR h R k wherein said alkyl, alkylene, cycloalkyl, and heterocyclyl are optionally substituted one or more times with substituents independently selected from R c ; or G is -CH2Y 3 , -CH2CH2Y 3 , -CH2CH2CH2Y 3 , -CH(CH3)CH2Y 3 , -CH2CH(Y 3 )CH3, -CH(Y3 )CH3, -CH2C(Y 3 )(CH3)2, -C(Y 3 )(CH3)2, or wherein Y 3 is cyclopropyl, -CF3, -OCF3, -OCH3, -OCH2CH3, -F, -Cl, -OH, -O(CH2)2-OH, -O(CH2)2-F, -SCH3, -S(O)2-CH3, -SCH2CH3, -S(O)2CH2CH3, -NH-CH3, -NH-CH2CH3, -N(CH3)2, tetrahydropyran-4-yl, tetrahydrofuran-2-yl, morpholin-2-yl, morpholin-4-yl, piperidin-1-yl, 4-hydroxy-piperidin-1-yl, 3-hydroxy-piperidin-1-yl, -NH-C(O)-CH3, -NH-C(O)-CH2CH3, tetrahydrofuran-2-yl-methyloxy, or -C(O)-Y 4 wherein Y 4 is -OH, -OCH3, -OCH2CH3, -OC(CH3)3, -NH2, -NH-CH3, -NH-CH2CH3, -N(CH3)2, -N(CH2CH3)2, morpholin-4-yl, 4-methyl-piperazin-1-yl, pyrrolidin-1-yl, or piperazin-1-yl;

[0516] L is -CH2-C(O)N(R 6 )-, -C(O)N(R 6 )-, -C(O)-O-, -SO2-, -C(O)-, or heterocyclylenyl, optionally substituted one or more times by substituents independently selected from R x ; or the group -L-G is -cyano;

[0517] R 1 is hydrogen or R a ;

[0518] R c is

[0519] a) halo,

[0520] b) -C 1-6 alkyl,

[0521] c) -C 3-10 cycloalkyl,

[0522] d) -heterocyclyl,

[0523] e) -cyano,

[0524] f) -CF3,

[0525] g) -OCF3,

[0526] h)-OR h ,

[0527] i)-S(O) w -R h ,

[0528] j)-S(O)2O-R h ,

[0529] k)-NR h R k ,

[0530] l)-C(O)-R h ,

[0531] m)-C(O)-OR h ,

[0532] n)-OC(O)-R h ,

[0533] o)-C(O)NR h R k ,

[0534] p)-C(O)-heterocyclic group,

[0535] q)-NR h C(O)R k ,

[0536] r)-OC(O)NR h R k ,

[0537] s)-NR h C(O)OR k ,

[0538] t)-NR h C(O)NR h R k ,

[0539] u)-NR h S(O) w R k ,or

[0540] v)-O-(C 1-4 alkylene)-O-(C 1-4 alkylene)-N(R h )C(O)-OR k ,

[0541] The alkylene, alkyl, cycloalkyl, and heterocyclic groups thereon are optionally selected independently from R xsubstituted one or more times with a substituent independently selected from R

[0542] R h and R k are independently hydrogen, C 1-6 alkyl, C 3-10 cycloalkyl, or heterocyclyl, wherein said alkyl, cycloalkyl, and heterocyclyl are optionally substituted one or more times with a substituent independently selected from R x ; or, if R h and R k are both attached to the same nitrogen atom, together with the nitrogen atom to which they are attached can optionally form a heterocyclic ring selected from azetidinyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, piperidinyl, piperazinyl, morpholino, thiomorpholino, and azepanyl, wherein each ring is optionally substituted one or more times with a substituent independently selected from R x ; and

[0543] R x is R y .

[0544] Embodiment 114: The compound according to any one of Embodiments 66 to 112, wherein

[0545] -L-G is not -cyano.

[0546] Embodiment 115: The compound according to any one of Embodiments 66 to 112, wherein

[0547] L is -C(O)N(R 6 )-.

[0548] Embodiment 116: The compound according to Embodiment 115, wherein

[0549] R 6 is hydrogen.

[0550] Embodiment 117: The compound according to Embodiment 115, wherein

[0551] R 6 is methyl.

[0552] Embodiment 118: The compound according to Embodiment 117, wherein

[0553] G is -N(CH3)2.

[0554] Embodiment 119: The compound according to any one of Embodiments 66 to 112, wherein

[0555] -L-G is -C(O)NR h R k .

[0556] Embodiment 120: The compound according to Embodiment 119, wherein

[0557] NR h R k is pyrrolidinyl, piperidinyl, piperazinyl, 4-methyl-piperazinyl, or morpholino, wherein each of the foregoing groups is optionally substituted with -(CH2) 1-3 -OH one time.

[0558] Embodiment 121 : The compound according to Embodiment 120, wherein

[0559] NR h R k is pyrrolidinyl, 4-(2-hydroxyethyl)-piperazinyl, or 4-(3-hydroxypropyl)-piperidinyl.

[0560] Embodiment 122: The compound according to Embodiment 119, wherein

[0561] NR h R k is N[(CH2)2-OH]2.

[0562] Embodiment 123: The compound according to any one of Embodiments 66 to 114, wherein

[0563] L is not -CH2-C(O)N(R 6 )-.

[0564] Embodiment 124: The compound according to any one of Embodiments 66 to 123, wherein

[0565] L is not heterocyclylenyl.

[0566] Embodiment 125: The compound according to any one of Embodiments 66 to 112, wherein

[0567] L is -S(O)2-.

[0568] Embodiment 126: The compound according to Embodiment 125, wherein

[0569] G is methyl or -CF3.

[0570] Embodiment 127: The compound according to any one of Embodiments 66 to 112, wherein

[0571] L is heteroarylenyl, optionally substituted one or more times with a substituent independently selected from R x .

[0572] Embodiment 128: The compound according to Embodiment 127, wherein

[0573] -L-G is imidazol-2-yl, 1,2,4-triazol-3-yl, or 5-methyl-l,2,4-triazol-3-yl.

[0574] Embodiment 129: The compound according to any one of Embodiments 66 to 112, wherein

[0575] L is -C(O)-O-.

[0576] Embodiment 130: The compound according to Embodiment 129, wherein

[0577] G is hydrogen, or -C 1-8 alkyl, wherein said alkyl is optionally substituted one or more times by substituents independently selected from R c

[0578] Embodiment 131: The compound according to Embodiment 130, wherein

[0579] G is methyl or ethyl.

[0580] Embodiment 132: The compound according to Embodiment 130, wherein

[0581] G is hydrogen.

[0582] Embodiment 133: The compound according to any one of Embodiments 66 to 116, wherein

[0583] G is -C 1-8 alkyl, -C 3-10 cycloalkyl, -C 1-6 alkylene-C 3-10 cycloalkyl, heterocyclyl, or -C 1-6 alkylene-C 3-10 heterocyclyl, wherein said alkyl, alkylene, cycloalkyl, and heterocyclyl are optionally substituted one or more times by substituents independently selected from R c

[0584] Embodiment 134: The compound according to Embodiment 133, wherein

[0585] G is -C 1-8 alkyl, optionally substituted one or more times by substituents independently selected from R c

[0586] Embodiment 135: The compound according to Embodiment 134, wherein

[0587] G is methyl, ethyl, isopropyl, n-propyl, n-butyl, sec-butyl, or isobutyl.

[0588] Embodiment 136: The compound according to Embodiment 134, wherein​​​

[0589] G is methyl, ethyl, or n-propyl.

[0590] Embodiment 137: The compound according to Embodiment 134, wherein

[0591] G is 2-fluoroethyl, 2,2-difluoroethyl, or 2,2,2-trifluoroethyl.

[0592] Embodiment 138: The compound according to Embodiment 134, wherein

[0593] G is 2-cyanoethyl.

[0594] Embodiment 139: The compound according to Embodiment 134, wherein

[0595] G is -G 1-8 alkyl, substituted once by -C(O)-O-R h .

[0596] Embodiment 140: The compound according to Embodiment 139, wherein

[0597] G is -CH2-C(O)-O-R h .

[0598] Embodiment 141: The compound according to Embodiment 140, wherein

[0599] R h is hydrogen or methyl.

[0600] Embodiment 142: The compound according to Embodiment 139, wherein

[0601] G is -CH2CH2-C(O)-O-R h .

[0602] Embodiment 143: The compound according to Embodiment 142, wherein

[0603] R h is hydrogen or methyl.

[0604] Embodiment 144: The compound according to Embodiment 139, wherein

[0605] G is -C(CH3)2-C(O)-O-R h .

[0606] Embodiment 145: The compound according to Embodiment 144, wherein

[0607] R h is hydrogen or methyl.

[0608] Embodiment 146: A compound according to Embodiment 139, wherein

[0609] G is -CH(CH3)-C(O)-O-R h .

[0610] Embodiment 147: A compound according to Embodiment 146, wherein

[0611] R h is hydrogen or methyl.

[0612] Embodiment 148: A compound according to Embodiment 134, wherein

[0613] G is -C 1-8 alkyl, substituted once with -C(O)NR h R k .

[0614] Embodiment 149: A compound according to Embodiment 148, wherein

[0615] G is CH2-C(O)-NR h R k .

[0616] Embodiment 150: A compound according to Embodiment 149, wherein

[0617] NR h R k is methylamino, dimethylamino, or diethylamino.

[0618] Embodiment 151: A compound according to Embodiment 149, wherein

[0619] NR h R k is thiomorpholino or 1,1-dioxothiomorpholino.

[0620] Embodiment 152: A compound according to Embodiment 149, wherein

[0621] NR h R k is morpholino, pyrrolidinyl, piperidinyl, piperazinyl, or 4-methylpiperazinyl.

[0622] Embodiment 153: A compound according to Embodiment 149, wherein

[0623] NR h R kis pyrrolidino, 3-hydroxy-pyrrolidino, 3-methoxy-pyrrolidino, 3-amino-pyrrolidino, 3-(methylamino)-pyrrolidino, 3-(dimethylamino)-pyrrolidino, 2-(hydroxymethyl)-pyrrolidino, 2-(dimethylaminocarbonyl)-pyrrolidino, or 3,4-dihydroxy-pyrrolidino.

[0624] Embodiment 154: The compound according to Embodiment 149, wherein

[0625] NR h R k is piperidino, 3-hydroxy-piperidino, 4-hydroxy-piperidino, 2-(hydroxymethyl)-piperidino, 3-(hydroxymethyl)-piperidino, 4-(hydroxymethyl)-piperidino, 3-methoxy-piperidino, 4-(methoxymethyl)-piperidino, 4-(fluoromethyl)-piperidino, 4-(trifluoromethyl)-piperidino, 4-cyano-piperidino, 4-carbamoyl-piperidino, 4-(methylamino)-piperidino, 4-(dimethylamino)-piperidino, 4-(methylaminomethyl)-piperidino, or 4-(dimethylaminomethyl)-piperidino.

[0626] Embodiment 155: The compound according to Embodiment 149, wherein

[0627] NR h R k is piperidino, 3-hydroxy-piperidino, 4-hydroxy-piperidino, 2-(hydroxymethyl)-piperidino, 3-(hydroxymethyl)-piperidino, 4-(hydroxymethyl)-piperidino, 3-methoxy-piperidino, 4-(methoxymethyl)-piperidino, 4-(fluoromethyl)-piperidino, 4-(trifluoromethyl)-piperidino, 4-cyano-piperidino, 4-carbamoyl-piperidino, 4-(methylamino)-piperidino, 4-(dimethylamino)-piperidino, 4-(methylaminomethyl)-piperidino, or 4-(dimethylaminomethyl)-piperidino.

[0628] Embodiment 156: The compound according to Embodiment 149, wherein

[0629] NR h R k is NHR k wherein R k is 2-hydroxypropyl, 2-(methylsulfonyl)-ethyl, tetrahydrofuran-3-yl, tetrahydropyran-4-yl, 1-methylpiperidin-4-yl, piperidin-3-yl, or 1-methylpiperidin-3-yl.

[0630] Embodiment 157: The compound according to Embodiment 149, wherein

[0631] NR h R k is N(CH3)R k wherein R k is 2-hydroxyethyl, tetrahydropyran-4-yl, pyrrolidin-3-yl, 1-methylpyrrolidin-3-yl, or piperazin-3-yl.

[0632] Embodiment 158: A compound according to Embodiment 149, wherein

[0633] NR h R k is N(CH2CH2OH)2.

[0634] Embodiment 159: A compound according to Embodiment 148, wherein

[0635] G is -(CH2) 2-3 -C(O)-N(CH3)2.

[0636] Embodiment 160: A compound according to Embodiment 148, wherein

[0637] G is -(CH2)3-C(O)-(4-methylpiperazino).

[0638] Embodiment 161: A compound according to Embodiment 148, wherein

[0639] G is -CH(CH3)-C(O)-NR h R k , wherein NR h R k is methylamino, dimethylamino, 4-methylpiperazino, or morpholino.

[0640] Embodiment 162: A compound according to Embodiment 148, wherein

[0641] G is -C(CH3)2-C(O)-N(CH3)2.

[0642] Embodiment 163: A compound according to Embodiment 134, wherein

[0643] G is -CH-[C(O)-N(CH3)2]-[CH2OH], -CH-[C(O)-N(CH3)2]-[(CH2)4-NH2], or -CH-[C(O)-N(CH3)2]-[(CH2)4-N(CH3)2].

[0644] Embodiment 164: A compound according to Embodiment 134, wherein

[0645] G is -C 1-8 alkyl, substituted once by -O-R h .

[0646] Embodiment 165: A compound according to Embodiment 164, wherein

[0647] G is -(CH2)2-O-R h .

[0648] Embodiment 166: The compound according to Embodiment 165, wherein

[0649] R h is hydrogen, methyl, or ethyl.

[0650] Embodiment 167: The compound according to Embodiment 165, wherein

[0651] R h is trifluoromethyl, 2-fluoroethyl, 3-fluoropropyl, or 2,2-difluoroethyl.

[0652] Embodiment 168: The compound according to Embodiment 165, wherein

[0653] R h is tetrahydrofuran-2-ylmethyl.

[0654] Embodiment 169: The compound according to Embodiment 165, wherein

[0655] R h is 2-hydroxyethyl.

[0656] Embodiment 170: The compound according to Embodiment 165, wherein

[0657] R h is 3-hydroxypropyl.

[0658] Embodiment 171: The compound according to Embodiment 165, wherein

[0659] R h is 2-methoxyethyl.

[0660] Embodiment 172: The compound according to Embodiment 165, wherein

[0661] R h is 2-(2-hydroxyethoxy)-ethyl.

[0662] Embodiment 173: The compound according to Embodiment 165, wherein

[0663] R h is 2-hydroxypropyl or 1-hydroxypropan-2-yl.

[0664] Embodiment 174: The compound according to Embodiment 165, wherein

[0665] R h is 2-cyanoethyl, 2-(methylcarbonylamino)-ethyl, or 2-(methylsulfonylamino)-ethyl.

[0666] Embodiment 175: The compound according to Embodiment 165, wherein

[0667] R h is 2-aminoethyl, 2-(methylamino)-ethyl, or 2-(dimethylamino)-ethyl.

[0668] Embodiment 176: The compound according to Embodiment 165, wherein

[0669] R h is carbamoylmethyl.

[0670] Embodiment 177: The compound according to Embodiment 164, wherein

[0671] G is -(CH2)3-O-R h .

[0672] Embodiment 178: The compound according to Embodiment 177, wherein

[0673] R h is hydrogen, methyl, or ethyl.

[0674] Embodiment 179: The compound according to Embodiment 177, wherein

[0675] R h is 2-hydroxyethyl.

[0676] Embodiment 180: The compound according to Embodiment 164, wherein

[0677] G is -(CH2)4-OH, -(CH2)5-OH, -CH2C(CH3)2-OH, -CH2C(CH3)2-OCH3, -CH2C(CH3)2-CH2-OH, -CH(CH3)-CH2-OCH3, -(CH2)3C(CH3)2-CH2-OH, -(CH2)2CH(CH3)-CH2-OH, or -(CH2)2CH(CH3)-OH.

[0678] Embodiment 181: The compound according to Embodiment 164, wherein

[0679] G is -CH2CH(CH3)-O-R h .

[0680] Embodiment 182: The compound according to Embodiment 181, wherein

[0681] R h is hydrogen, methyl, or ethyl.

[0682] Embodiment 183: The compound according to Embodiment 134, wherein

[0683] G is -CH2-CH(OH)-CH2-OH.

[0684] Embodiment 184: The compound according to Embodiment 134, wherein

[0685] G is -C 1-8 alkyl, by -NR h R k is substituted once.

[0686] Embodiment 185: The compound according to Embodiment 184, wherein

[0687] G is -(CH2)2-NR h R k .

[0688] Embodiment 186: The compound according to Embodiment 185, wherein

[0689] NR h R k is amino, methylamino, or dimethylamino.

[0690] Embodiment 187: The compound according to Embodiment 185, wherein

[0691] NR h R k is methylcarbonylamino.

[0692] Embodiment 188: The compound according to Embodiment 185, wherein

[0693] NR h R k is (dimethylamino)methylcarbonylamino, hydroxymethylcarbonylamino, or 1- hydroxyethylcarbonylamino.

[0694] Embodiment 189: The compound according to Embodiment 185, wherein

[0695] NR h R k is methylsulfonylamino.

[0696] Embodiment 190: The compound according to Embodiment 185, wherein

[0697] NR h R k is piperidino, 4-hydroxypiperidino, or 3-hydroxypiperidino.

[0698] Embodiment 191: The compound according to Embodiment 185, wherein

[0699] NR h R kis piperidino, 4,4-difluoropiperidino, or 3,3-difluoropiperidino.

[0700] Embodiment 192: The compound according to Embodiment 185, wherein

[0701] NR h R k is 2-oxo-pyrrolidino, 2-oxo-imidazolidino, or 3-oxo-piperazino.

[0702] Embodiment 193: The compound according to Embodiment 185, wherein

[0703] NR h R k is piperazino, 4-methylpiperazino, morpholino, or 1,1-dioxo-thiomorpholino.

[0704] Embodiment 194: The compound according to Embodiment 184, wherein

[0705] G is -(CH2)3-NR h R k .

[0706] Embodiment 195: The compound according to Embodiment 194, wherein

[0707] NR h R k is amino, dimethylamino, or diethylamino.

[0708] Embodiment 196: The compound according to Embodiment 194, wherein

[0709] NR h R k is piperidino, 4-methylpiperazino, or morpholino.

[0710] Embodiment 197: The compound according to Embodiment 184, wherein

[0711] G is -(CH2)4-NR h R k .

[0712] Embodiment 198: The compound according to Embodiment 197, wherein

[0713] NR h R k is amino, dimethylamino, or diethylamino.

[0714] Embodiment 199: The compound according to Embodiment 133, wherein

[0715] G is -C 1-6alkylene-heterocyclyl, wherein the alkylene and heterocyclyl are optionally substituted one or more times by substituents independently selected from R c

[0716] Embodiment 200: The compound according to Embodiment 199, wherein

[0717] G is -CH2-heterocyclyl, wherein the heterocyclyl is optionally substituted one time with a substituent selected from R c

[0718] Embodiment 201 : The compound according to Embodiment 200, wherein

[0719] the heterocyclyl is tetrahydropyran-4-yl, tetrahydrofuran-2-yl, 1,4-dioxan-2-yl, morpholin-2-yl, tetrahydropyran-2-yl, piperidin-4-yl, 1-(2-hydroxyethyl)-piperidin-4-yl, 1-(dimethylaminomethylcarbonyl)-piperidin-4-yl, piperazin-2-yl, or 1-methyl-piperazin-2-yl.

[0720] Embodiment 202: The compound according to Embodiment 133, wherein

[0721] G is C 3-10 cycloalkyl, optionally substituted one or more times by substituents independently selected from R c

[0722] Embodiment 203: The compound according to Embodiment 202, wherein

[0723] G is 4-hydroxy-cyclohexyl, 4-carboxy-cyclohexyl, or 4-(dimethylaminocarbonyl)-cyclohexyl.

[0724] Embodiment 204: The compound according to Embodiment 202, wherein

[0725] G is 1-carboxy-cyclopropyl, 1-(ethoxycarbonyl)-cyclopropyl, or 1-(dimethylamino- carbonyl)-cyclopropyl.

[0726] Embodiment 205: The compound according to Embodiment 133, wherein

[0727] G is C 1-6 alkylene-C 3-10 cycloalkyl, wherein the alkylene and cycloalkyl are optionally substituted one or more times by substituents independently selected from R c

[0728] Embodiment 206: The compound according to Embodiment 205, wherein

[0729] G is -CH2-(4-hydroxy-cyclohexyl). ​​​​

[0730] Embodiment 207: A compound according to Embodiment 205, wherein

[0731] G is -(CH2)2-(4-hydroxy-cyclohexyl).

[0732] Embodiment 208: A compound according to Embodiment 205, wherein

[0733] G is -CH2-[4-(hydroxymethyl)-cyclohexyl].

[0734] Embodiment 209: A compound according to Embodiment 133, wherein

[0735] G is heterocyclyl, optionally substituted one or more times by substituents independently selected from R c

[0736] Embodiment 210: A compound according to Embodiment 209, wherein

[0737] G is piperidin-4-yl, 1-methyl-piperidin-4-yl, 1-carboxy-piperidin-4-yl, 1- (methylsulfonyl)-piperidin-4-yl, 1-(2-hydroxyethyl)-piperidin-4-yl, 1- (dimethyl-aminocarbonyl)piperidin-4-yl, or 1-(dimethylaminomethylcarbonyl)- piperidin-4-yl.

[0738] Embodiment 211: A compound according to Embodiment 209, wherein

[0739] G is piperidin-3-yl or 1-(dimethylaminomethylcarbonyl)-piperidin-3-yl.

[0740] Embodiment 212: A compound according to Embodiment 209, wherein

[0741] G is 1,1-dioxo-tetrahydrothiophen-3-yl.

[0742] Embodiment 213: A compound according to Embodiment 209, wherein

[0743] G is pyrrolidin-3-yl, 1-methyl-pyrrolidin-3-yl, 1-(2-hydroxyethyl)-pyrrolidin-3-yl, 1- (2-hydroxypropyl)-pyrrolidin-3-yl, 1-(2-hydroxy-2-methylpropyl)-pyrrolidin-3-yl, 1- (1-hydroxyethylcarbonyl)-pyrrolidin-3-yl, 1-(2-carboxyethyl)-pyrrolidin-3-yl, or 1- (2-methylsulfonylamino-ethyl)-pyrrolidin-3-yl.

[0744] Embodiment 214: A compound according to Embodiment 134, wherein

[0745] G is -C 1-8 ​alkyl, substituted once by -S-R h substituted once.

[0746] Embodiment 215: The compound according to Embodiment 214, wherein

[0747] G is -(CH2)2-S-R h .

[0748] Embodiment 216: The compound according to Embodiment 215, wherein

[0749] R h is methyl or ethyl.

[0750] Embodiment 217: The compound according to Embodiment 215, wherein

[0751] R h is 2-hydroxyethyl.

[0752] Embodiment 218: The compound according to Embodiment 214, wherein

[0753] G is -(CH2)3-S-R h .

[0754] Embodiment 219: The compound according to Embodiment 218, wherein

[0755] R h is methyl.

[0756] Embodiment 220: The compound according to Embodiment 134, wherein

[0757] G is -C 1-8 alkyl, substituted once by -SO2-R h .

[0758] Embodiment 221: The compound according to Embodiment 220, wherein

[0759] G is -(CH2)2-SO2-R h .

[0760] Embodiment 222: The compound according to Embodiment 221, wherein

[0761] R h is methyl or ethyl.

[0762] Embodiment 223: The compound according to Embodiment 221, wherein

[0763] R h is 2-hydroxyethyl.

[0764] Embodiment 224: The compound according to Embodiment 220, wherein

[0765] G is -(CH2)3-SO2-R h .

[0766] Embodiment 225: The compound according to Embodiment 224, wherein

[0767] R h is methyl.

[0768] Embodiment 226: The compound according to Embodiment 133, wherein

[0769] G is -CH(CH3)-NR h R k , wherein NR h R k is pyrrolidinyl, piperidinyl, 4-methyl-piperazinyl, morpholino, or dimethylamino.

[0770] Embodiment 227: The compound according to Embodiment 133, wherein

[0771] G is 1-(2-hydroxypropyl)-pyrrolidin-3-yl or 1-(1-hydroxyethylcarbonyl)-pyrrolidin-3-yl.

[0772] Embodiment 228: The compound according to Embodiment 133, wherein

[0773] G is 1-(dimethylaminomethylcarbonyl)-piperidin-4-yl.

[0774] Embodiment 229: The compound according to Embodiment 133, wherein

[0775] G is -(CH2) 3-5 -OH.

[0776] Embodiment 230: The compound according to Embodiment 133, wherein

[0777] G is 4-hydroxy-cyclohexylmethyl.

[0778] Embodiment 231: The compound according to Embodiment 133, wherein

[0779] G is -(CH2)2-NHC(O)-CH2-N(CH3)2.

[0780] Embodiment 232: The compound according to Embodiment 133, wherein

[0781] G is 4-hydroxy-cyclohexylmethyl.

[0782] Embodiment 233: The compound according to Embodiment 133, wherein

[0783] G is -CH2-C(O)-NR h R k wherein NR h R k is 3-hydroxy-pyrrolidinyl or 3-(dimethyl-amino)-pyrrolidinyl.

[0784] Embodiment 234: The compound according to Embodiment 133, wherein

[0785] G is -CH2-C(O)-NR h R k wherein NR h R k is morpholino.

[0786] Embodiment 235: The compound according to Embodiment 133, wherein

[0787] G is -CH2-C(O)-NR h R k wherein NR h R k is 4-hydroxy-piperidinyl, 4-methoxy-piperidinyl, 4-(hydroxymethyl)-piperidinyl, 3- hydroxy-piperidinyl, 3-methoxy-piperidinyl, 3-(hydroxymethyl)-piperidinyl, or 4,4- difluoropiperidinyl.

[0788] Embodiment 236: The compound according to Embodiment 133, wherein

[0789] G is -CH2-C(O)-NR h R k wherein NR h R k is dimethylamino.

[0790] Embodiment 237: The compound according to Embodiment 133, wherein

[0791] G is -(CH2)2-O-(CH2)2-OH.

[0792] Embodiment 238: The compound according to Embodiment 133, wherein

[0793] G is -(CH2)2-O-(CH2)2-OCH3.

[0794] Embodiment 239: The compound according to Embodiment 133, wherein

[0795] G is -CH2-CH(CH3)-OH.

[0796] Embodiment 240: A compound according to any one of Embodiments 66 to 112, wherein

[0797] L is C(O)NH and G is C substituted once with heteroaryl 1-8 alkyl, wherein said heteroaryl is optionally substituted one or more times with a substituent independently selected from R x

[0798] Embodiment 241 : A compound according to Embodiment 240, wherein

[0799] G is -CH2-(2-furyl), -CH2-(2-thienyl), -CH2-(2-oxazolyl), or -CH2-(2-thiazolyl).

[0800] Embodiment 242: A compound according to Embodiment 240, wherein

[0801] G is -(CH2) 2-3 -(1-pyrrolyl), -(CH2) 2-3 -(1-pyrazolyl), or -(CH2) 2-3 -(1-imidazolyl).

[0802] Embodiment 243: A compound according to any one of Embodiments 66 to 112, wherein

[0803] L is C(O)NH and G is C substituted once with phenyl 1-8 alkyl, wherein said phenyl is optionally substituted one or more times with a substituent independently selected from R x

[0804] Embodiment 244: A compound according to Embodiment 243, wherein

[0805] G is -(-CH2) 1-2 -(4-hydroxyphenyl) or -(-CH2) 1-2 -(4-methoxy-3-hydroxyphenyl).

[0806] Embodiment 245: A compound according to any one of Embodiments 66 to 112, wherein

[0807] L is C(O)NH and G is -CH2-C(O)NH-CH2-(4-hydroxyphenyl).

[0808] Embodiment 246: A compound according to any one of Embodiments 66 to 112, wherein

[0809] L is C(O)NH and G is -CH2-C(O)-[4-(pyrimidin-2-yloxy)-piperidino].

[0810] ​​Embodiment 247: The compound according to any one of embodiments 1 to 246, wherein X 2 is =C(R 1 )- and X 3 is =C(-L-G)-.

[0811] Embodiment 248: The compound according to any one of embodiments 1 to 247, wherein

[0812] said compound is in free acid or free base form.

[0813] Embodiment 249: The compound according to any one of embodiments 1 to 247, wherein said compound is in pharmaceutically acceptable salt form.

[0814] Embodiment 250: The compound according to embodiment 1, wherein said compound is a compound from Table A or a pharmaceutically acceptable salt thereof.

[0815] Table A

[0816]

[0817]

[0818]

[0819]

[0820]

[0821]

[0822]

[0823]

[0824]

[0825]

[0826]

[0827]

[0828]

[0829]

[0830]

[0831]

[0832]

[0833]

[0834]

[0835]

[0836]

[0837]

[0838]

[0839]

[0840]

[0841]

[0842]

[0843]

[0844]

[0845]

[0846]

[0847]

[0848]

[0849]

[0850]

[0851]

[0852]

[0853]

[0854]

[0855]

[0856]

[0857]

[0858]

[0859]

[0860]

[0861] Compounds 1-474 in Table A can be prepared as described in WO'018 or other methods apparent to one skilled in the art. For example, compounds 473 and 474 in Table A can be prepared as described in the Examples section below.

[0862] In another aspect, the present application provides a pharmaceutical composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in treating sickle cell disease or related conditions. In embodiments, the present application provides a pharmaceutical composition and a pharmaceutical carrier comprising a compound (or salt) of any one of embodiments 1 to 250 (as described above). In embodiments, the pharmaceutical composition comprises a compound (or salt) of any one of embodiments 1 to 250 and a pharmaceutically acceptable carrier.

[0863] Thus, in yet another embodiment, the present application provides a pharmaceutical composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. In yet another embodiment, the present application provides a pharmaceutical composition comprising a compound (or salt) of any one of embodiments 1 to 250 and a pharmaceutically acceptable carrier.

[0864] In yet another embodiment, the present application provides a compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in medicine. In yet another embodiment, the present application provides a compound (or salt) of any one of embodiments 1 to 250 for use in medicine.

[0865] B. Co-administration

[0866] The present application also provides the use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, in combination with one or more pharmaceutically active compounds, for simultaneous, sequential or separate administration. The present application also provides the use of a compound (or salt) of any one of embodiments 1 to 250, in combination with one or more active compounds, for simultaneous, sequential or separate administration. Examples of the active ingredients include, but are not limited to, HU, Nrf2 activators, antioxidants, antidotes, and anti-inflammatory agents. In one embodiment, the present application provides a pharmaceutical composition comprising a compound (or salt) of any one of embodiments 1 to 250 and at least one other pharmaceutically active ingredient selected from HU, Nrf2 activators, antioxidants, antidotes, and anti-inflammatory agents. In yet another embodiment, the present application provides the use of a compound (or salt) of any one of embodiments 1 to 250, in combination with at least one other pharmaceutically active ingredient selected from Nrf2 activators, antioxidants, antidotes, and anti-inflammatory agents, for simultaneous, sequential or separate administration.

[0867] The Nrf2 activator can comprise a Michael addition acceptor, one or more fumaric acid esters (i.e., mono- and / or di-esters of fumaric acid that can be selected from monoalkyl fumarate and dialkyl fumarate) such as monomethyl fumarate, dimethyl fumarate, monoethyl fumarate, and diethyl fumarate, in addition to edaravone, methyl bardoloxol (2-cyano-3,12-dioxo- olean-1,9(11)-dien-28-oate methyl ester), isothiocyanates such as sulforaphane, 1,2- dithiol-3-thione such as oltipraz, 3,5-di-tert-butyl-4-hydroxytoluene, 3-hydroxycoumarin, or a pharmaceutically active derivative or analog of the above agents. In one embodiment, the Nrf2 activator used in combination with the compounds of the present application is methyl bardoloxol and a fumaric acid ester.

[0868] Nrf2 activator compounds can be classified according to their chemical structure: diphenols, Michael reaction acceptors, isothiocyanates, thiocarbamates, trivalent arsenicals, 1,2-dithiol-3-thiones, hydroperoxides, ortho-dithiols, heavy metals, and polyenes. In general, Nrf2 activators are chemically reactive because they can be electrophilic agents, substrates for glutathione transferases, and / or can modify sulfhydryl groups through alkylation, oxidation, or reduction.

[0869] In another embodiment, the Nrf2 activator is methyl bardoloxol and a dialkyl fumarate such as dimethyl fumarate and diethyl fumarate.

[0870] In another embodiment, the Nrf2 activator is selected from the group consisting of a chalcone derivative such as 2-trifluoromethyl-2'-methoxychalcone, auranofin, ebselen, 1,2-naphthoquinone, cinnamaldehyde, caffeic acid and its esters, curcumin, resveratrol, artemether, tert-butylhydroquinone and tert-butylquinone (tBHQ, tBQ), vitamins Kl, K2 and K3, menadione, fumaric acid esters (i.e. mono- and / or diesters of fumaric acid which can be selected from monoalkyl fumarates and dialkyl fumarates) such as monomethyl fumarate, dimethyl fumarate (DMF), monoethyl fumarate and diethyl fumarate, 2-cyclopentenone, etaniic acid and its alkyl esters, methyl bardolomol (2-cyano-3,12-dioxooleana-l,9(l l)-dien-28-oic acid methyl ester) (CDDO-Me, RTA 402), 2-cyano-3,12-dioxooleana-l,9(l l)-dien-28-oic acid ethyl ester, 2-cyano-3,12-dioxooleana-l,9(l l)-dien-28-oic acid (CDDO), 1 [2-cyano-3,12-dioxooleana-l,9(l l)-dien-28-oyl]imidazole (CDDO-Im), (2-cyano-N-methyl-3,12-dioxooleana-l,9(l l)-dien-28 amide (CDDO-methylamide, CDDO-MA), isothiocyanates such as sulforaphane, 1,2-dithiole-3-thione such as oltipraz, 3,5-di-tert-butyl-4-hydroxytoluene, 3-hydroxycoumarin, 4-hydroxynonenal, 4-oxononenal, malondialdehyde, (E)-2-hexenal, capsaicin, allicin, allyl isothiocyanate, 6-methylthiohexyl isothiocyanate, 7-methylthioheptyl isothiocyanate, sulforaphane, 8-methylthiooctyl isothiocyanate, a corticosteroid such as dexamethasone, 8-iso prostaglandin A2, pyruvic acid alkyl esters such as pyruvic acid methyl ester and pyruvic acid ethyl ester, oxalopropionic acid diethyl ester or oxalopropionic acid dimethyl ester, 2-acetamidopropenoate, 2-acetamidopropenoic acid methyl ester or 2-acetamidopropenoic acid ethyl ester, hypoestoxide, parthenolide, eriodictyol, 4-hydroxy-2-nonenal, 4-oxo-2 nonenal, geranial, zingerone, aurone, isoliquiritigenin, xanthohumol, 10-gingerol, eugenol, l'-acetoxychavicol acetate, allyl isothiocyanate, benzyl isothiocyanate, phenethyl isothiocyanate, 4-(methylthio)-3-butenyl isothiocyanate and 6-methylsulfinylhexyl isothiocyanate, ferulic acid and its esters such as ferulic acid ethyl ester and ferulic acid methyl ester, sophorone, 4-methylplumericin, imperatorin, hesperetin, limettin, bis[2-hydroxybenzylidene]propanone, alicylcurcuminoid, 4-bromoflavone, β-naphthoflavone, sappanone A, aurone and its corresponding indole derivatives such as benzylidene-indol-2-one, perillaldehyde, quercetin, fisetin, koparin, genistein,Tanshinone HA, BHA, BHT, PMX-290, AL-1, avicin D, gedunin, fisetin, and the tricyclic bis(cyano enone) TBE-31 [(+ / -)-(4bS,8aR,10aS)-10a-ethynyl-4-b,8,8-trimethyl-3,7-dioxo-3,4-b,7,8,8a,9,10,10a-octahydrophenalene-2,6-dicarboxylic acid].

[0871] In another embodiment, the Nrf2 activator is selected from the group consisting of carnosic acid, 2-naphthoquinone, cinnamaldehyde, caffeic acid and esters thereof, curcumin, resveratrol, artemether, tert-butylhydroquinone, vitamins K1, K2 and K3, fumaric acid esters (i.e. fumaric acid mono- and / or di-esters, preferably selected from the group consisting of fumaric acid monoalkyl esters and fumaric acid dialkyl esters) such as fumaric acid monomethyl ester, fumaric acid dimethyl ester, fumaric acid monoethyl ester and fumaric acid diethyl ester, isothiocyanates such as sulforaphane, 1,2-dithiole-3-thione such as oltipraz, 3,5-di-tert-butyl-4-hydroxytoluene, 3-hydroxycoumarin, 4-hydroxynonenal, 4-oxononenal, malondialdehyde, (E)-2-hexenal, capsaicin, allicin, allyl isothiocyanate, 6-methylthiohexyl isothiocyanate, 7-methylthioheptyl isothiocyanate, sulforaphane, 8-methylthiooctyl isothiocyanate, 8-iso-prostaglandin A2, pyruvic acid alkyl esters such as pyruvic acid methyl ester and pyruvic acid ethyl ester, oxalopropionic acid diethyl ester or oxalopropionic acid dimethyl ester, 2-acetamidopropenoate, 2-acetamidopropenoic acid methyl ester or 2-acetamidopropenoic acid ethyl ester, artemin oxide, helenalin, eriodictyol, 4-hydroxy-2-nonenal, 4-oxo-2-nonenal, geranial, bergaptan, sinensetin, isoliquiritigenin, xanthohumol, 10-gingerol, eugenol, 1'-acetoxychavicol acetate, allyl isothiocyanate, benzyl isothiocyanate, phenethyl isothiocyanate, 4-(methylthio)-3-butenyl isothiocyanate and 6-methylsulfinylhexyl isothiocyanate, and the corresponding quinone or hydroquinone forms of the above quinone and hydroquinone derivatives.

[0872] In another embodiment, the Nrf2 activator can be a Michael reaction acceptor, such as dimethyl fumarate, fumaric acid monomethyl isothiocyanate, and 1,2-dithiole-3-thione. In another embodiment, the Nrf2 activator is selected from fumaric acid monomethyl ester, dimethyl fumarate, oltipraz, 1,2-naphthoquinone, tert-butylhydroquinone, methyl or ethyl pyruvate, 3,5-di-tert-butyl-4-hydroxytoluene, diethyl or dimethyl oxalylate, gossypolone, parthenolide, eriodicytol, 4-hydroxy-2-nonenal, 4-oxo-2-nonenal, geranial, zingiberone, aurone, isoliquiritigenin, xanthohumol, 10-gingerol, eugenol, 1'-acetoxychavicol acetate, allyl isothiocyanate, benzyl isothiocyanate, phenethyl isothiocyanate, 4-(methylthio)-3-butenyl isothiocyanate, and 6-methylsulfinylhexyl isothiocyanate.

[0873] Examples of antioxidants include vitamin C, vitamin E, carotenoids, retinolds, polyphenols, flavonoids, lignans, selenium, butylated hydroxyanisole, ethylenediaminetetraacetate, calcium disodium edetate, acetylcysteine, probucol, and tempo.

[0874] Examples of antidotes include dimethyl caprol, glutathione, acetylcysteine, methionine, sodium bicarbonate, deferoxamine mesylate, calcium disodium edetate, penicillamine hydrochloride, and medicinal charcoal.

[0875] Anti-inflammatory agents include steroidal anti-inflammatory agents and non-steroidal anti-inflammatory agents. Examples of steroidal anti-inflammatory agents include cortisone acetate, hydrocortisone, paramethasone acetate, prednisolone, prednisolone, methylprednisolone, dexamethasone, triamcinolone, and betamethasone. Examples of non-steroidal anti-inflammatory agents include salicylic acid non-steroidal anti-inflammatory agents such as aspirin, diflunisal, aspirin + vitamin C, and aspirin di-aluminum salt; aryl acid non-steroidal anti-inflammatory agents such as diclofenac sodium, sulindac, fenbufen, indomethacin, indomethacin fanel, acemetacin, proglumetacin maleate, anfenac sodium, nabmeton, mofebutazone, and etodorag; fenamic acid non-steroidal anti-inflammatory agents such as mefenamic acid, flufenamic acid aluminum, tolfenamic acid, and flufenisal; propionic acid non-steroidal anti-inflammatory agents such as ibuprofen, flurbiprofen, ketoprofen, naproxen, pranoprofen, fenoprofen calcium, thiaprofen, oxaprozin, loxoprofen sodium, alminoprofen, and zaltoprofen; oxicam non-steroidal anti-inflammatory agents such as piroxicam, ampiroxicam, tenoxicam, lomoxicam, and meloxicam; and basic non-steroidal anti-inflammatory agents such as tiaramide hydrochloride, epirizole, and emorfazone.

[0876] The physician can select a suitable time course for sequential administration based on factors such as the nature of the patient's disease and the patient's individual active agent dosing status. In certain embodiments, sequential administration comprises co-administration of one or more additional active agents within a period of one week, 72 hours, 48 hours, 24 hours, or 12 hours.

[0877] In some embodiments, the compositions disclosed herein are co-administered with one or more additional active agents for treating sickle cell disease, beta-thalassemia, or related conditions. Such additional active agents can include, but are not limited to, folic acid, penicillin or other antibiotics, preferably quinolones or macrolides, antiviral agents, antimalarial prophylactic agents, and analgesics to control pain crises.

[0878] In some embodiments, the compositions are co-administered with one or more additional agents that increase HbF expression, such as hydroxyurea (HU).

[0879] In some embodiments, the compositions are co-administered with one or more additional therapeutic regimens, for example, blood transfusion therapy, stem cell therapy, gene therapy, bone marrow transplantation, dialysis or kidney transplantation for kidney disease, cholecystectomy for gallstone patients, hip replacement for avascular necrosis of the hip, surgery for eye problems, and wound care for leg ulcers.

[0880] C. Effective Amount

[0881] In some embodiments, the compositions are administered in an amount effective to induce a pharmacological, physiological, or molecular effect, as compared to a control that is not administered the composition. In some embodiments, the compositions are administered to a subject in need thereof to increase expression of HbF in the subject.

[0882] Suitable controls are known in the art and can be determined based on the disease to be treated. Suitable controls include, but are not limited to, one or more subjects that do not have sickle cell disease, beta-thalassemia, or a sickle cell-related condition; or the condition or state of a subject that has the disease or condition prior to the start of treatment.

[0883] D. Dosage and Dosage Regimen

[0884] The dosage selected will depend on the therapeutic effect desired, the route of administration, and the duration of treatment desired. Generally, a dosage level of 0.001 to 100 mg / kg body weight per day is administered to a mammal. Generally, intravenous injection or infusion, the dosage can be lower.

[0885] Suitable dosages of a compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in the present application can be determined according to any of several recognized protocols. For example, animal studies, such as studies using mice, rats, dogs, and / or monkeys, can be used to determine appropriate dosages of a pharmaceutical compound. The results of animal studies can be extrapolated to determine dosages for other species, e.g., humans.

[0886] A compound of Formula (I) or a pharmaceutically acceptable salt thereof can be administered in a daily dose of 0.1 mg / kg to 15 mg / kg. In another embodiment, when the subject is a human, the daily dose can be between 1 mg and 1000 mg. In another embodiment, a compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered in an amount of 10 mg / day to 1000 mg / day, or 25 mg / day to 800 mg / day, or 37 mg / day to 750 mg / day, or 75 mg / day to 700 mg / day, or 100 mg / day to 600 mg / day, or 150 mg / day to 500 mg / day, or 200 mg / day to 400 mg / day. In other embodiments, each of the aforementioned daily administration periods of an amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof can be varied to every 6 hours, 12 hours, 48 hours, 72 hours, 96 hours, 1 week, or 2 weeks.

[0887] In some embodiments, a composition comprising a fumarate such as DMF, MMF, or a combination thereof, the daily dose of fumarate in a human can range from about 1 mg to about 5,000 mg, about 10 mg to about 2,500 mg, or about 50 mg to about 2,000 mg of fumarate or a pharmaceutically active salt thereof. In another embodiment, an effective dose of, for example, oral DMF or MMF, can be administered to a subject can be about 0.1 g to about 1 g or greater than 1 g per day; about 200 mg to about 800 mg per day; about 240 mg to about 720 mg per day; about 480 mg to about 720 mg per day; or about 720 mg per day. The daily dose can be administered in 2, 3, 4, or 6 equal divided administrations. In some embodiments, one or more fumarates or pharmaceutically active salts, derivatives, analogs, or prodrugs thereof are present in a pharmaceutical formulation. In some embodiments, the composition is administered to the patient three times per day (TID). In some embodiments, the pharmaceutical formulation is administered to the patient twice per day (BID). In some embodiments, the composition is administered to the patient at least one hour before or after the patient consumes food.

[0888] In some embodiments, the composition is administered as part of a dosing regimen. For example, a first dose of the composition can be administered to the patient during a first dosing period; and a second dose of the composition is administered during a second dosing period, followed by, optionally, one or more additional doses during one or more additional dosing periods. The first dosing period can be less than one week, equal to one week, or more than one week.

[0889] In some embodiments, the dosage regimen is a dose-escalating dosage regimen. The first dose can be a low dose, followed by measuring the HbF expression level, followed by steps of decreasing, maintaining or increasing the dose.

[0890] Current labeled dosing of hydroxyurea for sickle cell disease requires an initial dose of 15 mg / kg / day in a single dose and monitoring of the patient's blood cell counts every 2 weeks. If the blood cell counts are within acceptable ranges, the dose can be increased by 5 mg / kg / day every 12 weeks until the MTD of 35 mg / kg / day is reached. The pharmaceutical composition can comprise 1 mg / kg to 50 mg / kg of a fumarate such as MMF, and 1 mg / kg to 35 mg / kg of HU. The combination formulation can contain 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 mg / kg of HU.

[0891] E. Formulations

[0892] Pharmaceutical compositions comprising a compound of the present application are disclosed. The pharmaceutical compositions can be administered by oral, parenteral (intramuscular, intraperitoneal, intravenous (IV) or subcutaneous injection), transdermal (passively or using iontophoresis or electroporation) or transmucosal (nasal, vaginal, rectal or sublingual) routes of administration or using a bioerodible insert, and can be formulated into unit dosage forms suitable for each route of administration.

[0893] Erythrocytes are erythroid cells, which are the major producers of hemoglobin. Thus, in one embodiment, a compound or pharmaceutical composition of the present application is administered to a subject in an amount effective to induce HbF in hematopoietic stem cells. Thus, in some embodiments, a compound or pharmaceutical composition of the present application is administered in an amount effective to induce HbF expression in erythroid cells of the bone marrow (i.e., red bone marrow), liver, spleen, or a combination thereof.

[0894] In another embodiment, a compound or pharmaceutical composition of the present application induces HbF in cells that synthesize or contribute to the synthesis of hemoglobin. For example, in a preferred embodiment, a compound of the present application induces HbF in basophilic normoblasts / early normoblasts (also commonly referred to as erythroblasts), polychromatic normoblasts / intermediate normoblasts, orthochromatic normoblasts / late normoblasts, or a combination thereof.

[0895] In some embodiments, the compound or pharmaceutical composition of the application is administered locally to the site in need of treatment. While red blood cells are the primary producers of hemoglobin, other non-hematopoietic cells, including macrophages, retinal pigment cells, and alveolar epithelial cells (e.g., alveolar type II (ATII) cells) and Clara cells, can also synthesize hemoglobin. Thus, in some embodiments, the compound or pharmaceutical composition of the application is administered locally to an interface where oxygen-carbon dioxide diffusion occurs, including but not limited to the eye or the lung.

[0896] In some embodiments, the compound or pharmaceutical composition of the application is administered locally to the eye to treat retinal disease or another ocular manifestation associated with sickle cell disease or related conditions.

[0897] In one embodiment, the pharmaceutical composition is formulated for oral delivery. Oral solid dosage forms are described generally in Remington's Pharmaceutical Sciences, 21st ed., 2005, Chapter 45. Solid dosage forms include tablets, capsules, pills, troches or lozenges, cachets, pellets, powders or granules, or particulate formulations that incorporate the material into polymeric compounds, such as polylactic acid, polyglycolic acid, and the like, or into liposomes. Such compositions can affect the physical state, stability, rate of in vivo release, and rate of in vivo clearance of the disclosed. The composition can be prepared in liquid form, or can be prepared in dry powder (e.g., lyophilized) form. Another embodiment provides liquid dosage forms for oral administration including pharmaceutically-acceptable emulsions, solutions, suspensions, and syrups, which can contain other components including inert diluents; wetting agents, emulsifying agents, suspending agents, and the like adjuvants; and sweetening, flavoring, and perfuming agents.

[0898] Controlled release oral formulations can be desirable. The compounds of the application can be incorporated into inert matrices (e.g., gums) that allow release by diffusion or leaching mechanisms. Slowly degenerating matrices can also be incorporated into the formulation.

[0899] For oral formulations, the site of release can be the stomach, the small intestine (duodenum, jejunum, or ileum), or the large intestine.

[0900] IV. Therapeutic Methods

[0901] The treatment methods disclosed herein can include a first step of selecting a subject for treatment. In some embodiments, the subject is selected for treatment when the subject exhibits one or more clinical symptoms of, for example, sickle cell disease, beta-thalassemia, or a related disorder as discussed above. In some embodiments, the subject is selected for treatment when the subject exhibits a genetic or biochemical indicator of sickle cell disease, beta-thalassemia, or a related disorder. For example, the subject can be selected for treatment based on the identification of a genetic alteration, defect, or mutation in the beta-globin gene or its expression control sequences, by a biochemical or morphological alteration in hemoglobin or hemoglobin-synthesizing cells, or a combination thereof.

[0902] In some embodiments, the subject is selected when a combination of clinical symptoms and genetic or biochemical alterations are identified. In some embodiments, the subject is selected based on one or more clinical symptoms, or one or more genetic or biochemical alterations. For example, the subject can be selected for treatment based on the identification of a genetic alteration, biochemical or morphological alteration, or a combination thereof, prior to the subject exhibiting clinical symptoms of sickle cell disease, beta-thalassemia, or a related disorder.

[0903] In some embodiments, the treatment method can further include determining whether the subject is at risk of, or has, sickle cell disease, beta-thalassemia, or a related disorder by obtaining or having obtained a biological sample from the subject and performing or having performed a humoral test on the biological sample to determine whether the subject has one or more biomarkers or genetic mutations associated with sickle cell disease, beta-thalassemia, or a related disorder. If the subject is determined to be at risk of, or has, sickle cell disease, beta-thalassemia, or a related disorder, the method further includes administering to the subject a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

[0904] In any of the foregoing methods, the method can further include obtaining or having obtained a biological sample from the subject over a period of time and performing or having performed a humoral test on the biological sample to determine whether the level of one or more biochemical markers is increasing or decreasing, and administering a larger dose of the compound of Formula (I), or a pharmaceutically acceptable salt thereof, if the level of the one or more biochemical markers is not progressing in the desired direction. For example, the ratio of HbF to HbS in the sample can be measured, and a significant increase in the amount of HbF relative to HbS in a second sample from the subject relative to a first sample indicates that the dose of Formula (I), or a pharmaceutically acceptable salt thereof, is a therapeutically effective dose. Conversely, no change or no significant change in the amount of HbF relative to HbS in a second sample from the subject relative to a first sample can indicate that the dose of Formula (I), or a pharmaceutically acceptable salt thereof, is not a therapeutically effective dose and that an increase in dose can be needed.

[0905] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered to a subject in need thereof in an amount that reduces the level of one or more biomarkers, such as CRP or ROS.

[0906] The time interval over which the biological sample is collected can be 1 week, 2 weeks, 3 weeks, 4 weeks, 2 months, 3 months, 6 months, 9 months, or 12 months, and the compound of Formula (I), or a pharmaceutically acceptable salt thereof, can be administered during this period.

[0907] A. Identification of Genetic Alterations

[0908] In some embodiments, a subject is selected for treatment based on the identification of one or more genetic alterations in one or more alleles of the human beta-globin gene or expression control sequences thereof. Genetic alterations indicative of sickle cell disease, beta-thalassemia, or related conditions include the exemplary mutations discussed above, or other mutations that result in a decrease in the synthesis, structure, or function of human beta-globin.

[0909] Methods of selecting a subject having one or more genetic alterations in one or more alleles of the beta-globin gene or expression control sequences thereof include the steps of obtaining a biological sample and detecting the presence or absence of one or more genetic alterations. In one embodiment, the biological sample obtained contains nucleic acid from the subject, and the detecting step detects the presence or absence of one or more genetic alterations in one or more alleles of the beta-globin gene or expression control sequences thereof in the biological sample. Any biological sample containing DNA from the subject to be diagnosed can be used, including tissue samples and blood samples, with nucleated blood cells being a particularly convenient source. The DNA can be isolated from the biological sample prior to testing for the presence of genetic alterations.

[0910] The detecting step can include determining whether the subject is heterozygous or homozygous for the genetic alteration. The step of detecting the presence or absence of the genetic alteration can include a step of detecting whether the alteration is present in both chromosomes of the subject (i.e., detecting the presence or absence of one or both alleles containing the marker or functional polymorphism). More than one copy of the genetic alteration (i.e., a genetic marker in a homozygous subject) can indicate a greater risk of having sickle cell disease, beta-thalassemia, or related conditions. In some embodiments, the subject is heterozygous (also referred to herein as double heterozygous, triple heterozygous, etc.) for two or more genetic alterations in the beta-globin gene. One copy of two or more genetic alterations in the beta-globin gene can indicate a greater risk of having sickle cell disease, beta-thalassemia, or related conditions.

[0911] The process of determining the genetic sequence of the human beta-globin gene is referred to as genotyping. In some embodiments, the human beta-globin gene is sequenced. Methods of amplifying DNA fragments and sequencing them are well known in the art. For example, automated sequencing procedures that can be used to sequence the beta-globin gene include, but are not limited to, sequencing by mass spectrometry, single molecule real-time sequencing, ion semiconductor (ion torrent sequencing), pyrosequencing (454), sequencing by synthesis, sequencing by ligation, chain termination (Sanger sequencing).

[0912] In some embodiments, the genotype of the subject is determined by identifying the presence of one or more single nucleotide polymorphisms (SNPs) associated with sickle cell disease, beta-thalassemia, or related disorders. Methods of SNP genotyping are well known in the art. SNP genotyping can include the steps of collecting a biological sample (e.g., a sample of tissue, cells, bodily fluid, excretion, etc.) from the subject, isolating genomic DNA from cells of the sample, contacting the nucleic acid with one or more primers that specifically hybridize to a region of the isolated nucleic acid containing the target SNP under conditions that allow hybridization and amplification of the target nucleic acid region, and determining the nucleotide present at the SNP position of interest, or, in some assays, detecting the presence or absence of an amplified product (assays can be designed such that hybridization and / or amplification only occurs in the presence or absence of a particular SNP allele). In some assays, the size of the amplified product is detected and compared to the length of a control sample; for example, deletions and insertions can be detected by a change in the size of the amplified product compared to a normal genotype.

[0913] Flanking sequences can be used to design SNP detection reagents, such as oligonucleotide probes and primers. Common SNP genotyping methods include, but are not limited to, TaqMan analysis, molecular beacon analysis, nucleic acid arrays, allele-specific primer extension, allele-specific PCR, array primer extension, homogeneous primer extension analysis, primer extension detected by mass spectrometry, pyrosequencing, multiplex primer extension sorted on a gene array, rolling circle amplification ligation, homogeneous ligation, multiplex ligation reactions sorted on a gene array, restriction fragment length polymorphism, single base extension-tag analysis, and invasion analysis. Such methods can be used in conjunction with detection mechanisms such as luminescence or chemiluminescence detection, fluorescence detection, time-resolved fluorescence detection, fluorescence resonance energy transfer, fluorescence polarization, mass spectrometry, and electrochemical detection.

[0914] Other suitable methods for detecting polymorphisms include methods in which protection against cleavage agents is used to detect mismatched bases in RNA / RNA or RNA / DNA duplexes, comparison of electrophoretic mobility of variant and wild-type nucleic acid molecules, and use of denaturing gradient gel electrophoresis (DGGE) analysis to analyze the movement of polymorphic or wild-type fragments in polyacrylamide gels containing a gradient of denaturant. Sequence variations at particular positions can also be analyzed by nuclease protection assays (e.g., Rnase and S1 protection) or chemical cleavage methods.

[0915] Another method for genotyping SNPs uses two oligonucleotide probes in an oligonucleotide ligation assay (OLA). Other methods that can be used to genotype SNPs include single-strand conformation polymorphism (SSCP).

[0916] B. Identification of biochemical and morphological changes

[0917] In some embodiments, a subject is selected for treatment based on the identification of a biochemical or morphological change or abnormality in hemoglobin or a hemoglobin-synthesizing cell (e.g., a hematopoietic stem cell, a erythroid progenitor cell, a red blood cell, a macrophage, a retinal pigment epithelial cell, an alveolar type II (ATII) cell, etc.). This method generally includes identifying one or more biochemical or morphological changes associated with a genetic alteration in the human beta-globin gene or otherwise diagnostic of sickle cell disease, beta-thalassemia, or a related disorder. Methods for diagnosing sickle cell disease, beta-thalassemia, or a related disorder based on biochemical or morphological changes in hemoglobin or a hemoglobin-synthesizing cell are known in the art, including, but not limited to, red blood cell morphology analysis, osmotic fragility, hemoglobin composition, globin synthesis rate, and red blood cell indices.

[0918] In some embodiments, the method comprises first testing the subject’s blood for HbS, and selecting the subject for treatment if HbS is present. Methods of testing a subject’s blood for the presence of HbS include solubility tests (e.g., SICKLEDEX) and sickling tests. For SICKLEDEX testing, if HbS is present in the sample, it becomes insoluble and forms a cloudy suspension. Other hemoglobins are more soluble and will form a clear solution. Sickling tests can be used to determine if red blood cells become sickled after mixing a blood sample with a reducing agent, and identify morphological changes in red blood cell shape (i.e., “sickling”) by microscopy. Red blood cell shape changes can also be analyzed using a flow cytometer, such as an Amnis ImageStreamX Mark II imaging flow cytometer (MilliporeSigma). Software programs, such as the IDEAS application software (MilliporeSigma), can be used to quantify red blood cell shape changes using a modified protocol as described in: “Imaging flow cytometry for automated detection of hypoxia-induced erythrocyte shape change in sickle cell disease.” van Beers EJ, et al. Am J Hematol. 2014;89(6):598-603; or “Sickle Cell Imaging Flow Cytometry Assay (SIFCA).” Fertrin KY, et al. Methods Mol Biol. 2016;1389:279-292.

[0919] Other suitable tests include hemoglobin electrophoresis, which employs gel electrophoresis techniques to separate various types of hemoglobin from a blood sample obtained from a subject. This test can detect HbS as well as abnormal levels of other abnormal hemoglobins such as hemoglobin C. It can also be used to determine if there is a deficiency of any of the normal forms of hemoglobin such as in various thalassemias. Alternative methods of electrophoresis include isoelectric focusing and chromatographic techniques. Other tests that can be used to select a subject for treatment with the compositions and methods disclosed herein include tests that are typically used as part of a hemoglobinopathy screening, such as a complete blood count (CBC) or iron studies (ferritin). For example, a blood cell count can be used to detect anemia, and a blood smear can be used to identify sickle cells.

[0920] EMBODIMENT

[0921] General Procedure A: Ipso substitution of 6-chloro-5-nitro-nicotinic acid methyl ester

[0922] To a solution of 6-chloro-5-nitro-nicotinic acid methyl ester in DMF or THF was added a 2M solution of methylamine in THF and the reaction mixture was stirred at room temperature for 16 hours. The resulting mixture was poured into water to precipitate the product. The precipitate can be filtered and dried to give the product which can be used without further purification in the next step.

[0923] General Procedure B: Reduction of nitro group to amine

[0924] To a solution of the nitro compound in methanol was added 10% Pd / C. The resulting mixture was stirred at room temperature under an atmosphere of H2for 16 hours. The contents can then be filtered through a pad of celite or silica gel and the solids washed with portions of methanol. The filtrate and washings were combined and evaporated to provide the corresponding diamine which can be used without further purification in the next step.

[0925] General Procedure C: Formation of thiourea and its conversion to 2-amino- imidazopyridine

[0926] To a solution of the amine in acetonitrile (10 mL) was added 1,1'-thiocarbonylimidazole and the reaction mixture was stirred at room temperature (1-24 hours). The solvent was then evaporated and the product was suspended in acetonitrile. The solvent was then evaporated to yield the product as a precipitate. The precipitate was filtered and washed with acetonitrile and dried. The product was used directly in the next step without further purification.

[0927] To the product obtained immediately above was added EDAC at room temperature, followed by the substituted diamino pyridine and the reaction mixture was stirred at 90°C for 16 hours. The reaction mixture was then cooled to room temperature, poured into cold water and the solid was collected by filtration. The crude product thus obtained can be purified by trituration with methanol.

[0928] General Procedure D: Hydrolysis of ester

[0929] To a solution of the ester in 1:1 THF / MeOH was added aqueous NaOH and the resulting mixture was stirred at 60°C for 16 hours. After the reaction was complete, the mixture was concentrated under vacuum. The pH of the resulting suspension can be adjusted to pH ~3 by the dropwise addition of 6N HCl and the precipitate was collected by filtration, washed with water and dried under vacuum. The desired carboxylic acid was used without purification.

[0930] General Procedure E: Formation of amide using HBTU as coupling agent

[0931] To a solution of the carboxylic acid in anhydrous DMF was added DIEA, followed by HBTU, and the reaction mixture was stirred at room temperature for 30 minutes. The appropriate amine was then added, and the reaction was stirred at room temperature for 16 hours. The contents could be diluted with ice water, and the product precipitated. The product could be isolated after filtration with a subsequent wash with water and DCM / methanol or by silica gel chromatography using hexanes / ethyl acetate (80:20 to 60:40) as the eluent system.

[0932] Compound 473

[0933] N-[2-(2-hydroxyethoxy)ethyl]-3-methyl-2-[[6-(trifluoromethyl)-l,3-benzothiazol-2- yl]amino]imidazo[4,5-b]pyridine-6-carboxamide

[0934]

[0935] Following general procedure A, starting from 6-chloro-5-nitro-nicotinic acid methyl ester (5.0 g) and methylamine (33% in ethanol, 24 mL) in THF (150 mL), 6-methylamino-5-nitro-nicotinic acid methyl ester (5.0 g) was prepared. The crude product was used in the next step without further purification.

[0936] Following general procedure B, starting from 6-methylamino-5-nitro-nicotinic acid methyl ester (5.0 g) and Pd / C (20% by weight, 1.0 g) in methanol:THF (1:1, 50 mL), 5-amino-6-methylamino-nicotinic acid methyl ester (4.8 g) was prepared. The crude product was used in the next step without further purification.

[0937] Following general procedure C, starting from 6-(trifluoromethyl)-l,3-benzothiazol-2- amine (5.0 g), 5-amino-6-methylamino-nicotinic acid methyl ester (5.0 g), l,l'-thiocarbonyl- diimidazole (5.0 g), and EDAC (4.5 g), 3-methyl-2-[[6-(trifluoromethyl)-l,3-benzothiazol-2- yl]amino]imidazo[4,5-b]pyridine-6-carboxylic acid methyl ester (5.0 g) was prepared. The crude product was used in the next step without further purification.

[0938] Following general procedure D, starting from 3-methyl-2-[[6-(trifluoromethyl)-l,3- benzothiazol-2-yl]amino]imidazo[4,5-b]pyridine-6-carboxylic acid methyl ester (5.0 g) and NaOH (2 N, 25 mL) in methanol:THF (2:1, 50 mL), 3-methyl-2-[[6-(trifluoromethyl)-l,3- benzothiazol-2-yl]amino]imidazo[4,5-b]pyridine-6-carboxylic acid (4.2 g) was prepared. The crude product was used in the next step without further purification.

[0939] N-[2-(2-Hydroxyethoxy)ethyl]-3-methyl-2-[[6-(trifluoromethyl)-l,3-benzothiazol-2- yl]amino]imidazo[4,5-b]pyridine-6-carboxamide (40 mg) was prepared according to general procedure E, starting from 3-methyl-2-[[[6-(trifluoromethyl)-l,3-benzothiazol-2- yl]amino]imidazo[4,5-b]pyridine-6-carboxylic acid (100 mg), 2-(2- aminoethoxy)ethanol (100 mg), HBTU (200 mg) and DIEA (0.2 mL) in DMF (2.0 mL). LC / MS: m / z 481.7. 1 HNMR (DMSO-d6, 400 MHz): δ 8.67-8.59 (m, 2H), 8.29-8.23 (d, 2H), 7.71-7.69 (d, IH), 4.61 (s, IH), 3.68 (br s, 3H), 3.57-3.44 (m, 8H), 3.32 (br s, 2H).

[0940] Compound 474

[0941] 1-Ethyl-N-[2-(2-hydroxyethoxy)ethyl]-2-[[5-(trifluoromethoxy)-l,3-benzothiazol-2- yl]amino]benzoimidazole-5-carboxamide

[0942]

[0943] 1-Ethyl-N-[2-(2-hydroxyethoxy)ethyl]-2-[[5-(trifluoromethoxy)-l,3-benzothiazol-2- yl]amino]benzoimidazole-5-carboxamide (40 mg) was prepared according to general procedure E, starting from 3-ethyl-2-[[6-(trifluoromethoxy)-l,3-benzothiazol-2- yl]amino]imidazo[4,5-b]pyridine-6-carboxylic acid (100 mg) (see WO'018), 2-(2- aminoethoxy)ethanol (100 mg), HBTU (200 mg) and DIEA (0.2 mL) in DMF (2.0 mL). LC / MS: m / z 510.7.

[0944] General analytical methods

[0945] Western blot analysis

[0946] Approximately 20-50 pg of protein was separated by SDS-polyacrylamide gel electrophoresis and transferred onto a nitrocellulose membrane. The membrane was blocked in 5% milk powder with TBS-T for 30 minutes and then incubated with HbF or actin antibodies for 1 hour. After multiple washes, the membrane was incubated with a 1 : 10000 dilution of HRP-labelled secondary antibody (Thermo Scientific) and developed with ECL Prime reagent (GE Healthcare Bio-sciences). Images can be captured on a Bio-Rad Chemi-Doc MP imaging system and protein bands quantified by densitometry.

[0947] Flow cytometry assay

[0948] Approximately 5 x 105cells were harvested after treatment with compounds, washed twice with ice-cold phosphate buffered saline and resuspended in 4% paraformaldehyde for 40 minutes at 37°C. Cells were permeabilisation fixed with ice-cold acetone / methanol (4:1), washed with phosphate buffered saline and then incubated with FITC-labelled anti-HbF antibody (1 : 1000, Abeam) for 20 minutes. Labeled cells can be analysed using a Becton Dickerson LSR-II flow cytometer (BD Bioscience, San Jose, CA, USA) and FlowJo v0.9 software.

[0949] Example 1

[0950] KU812 is a human leukaemia cell line that expresses both fetal gamma-globin and adult beta-globin genes and was used as a screening system. KU812 cells have a globin gene response pattern comparable to primary red blood cells after treatment with potential HbF inducers. (See Zein S, Lou RF, Sivanand S, Ramakrishnan V, Mackie A, Li W, Pace BS. KU812 Cell Line: model for identifying fetal hemoglobin inducing drugs. Exp Biol Med (Maywood) 235: 1385-94, 2010.) KU812 cells were grown in Iscove’s Modified Dulbecco Media (IMDM) and 10% fetal bovine serum until log phase growth.

[0951] KU812 cells in log phase growth were treated with compounds 73, 134, 473 and 236 (see Table A) at doses of 0.5, 2.5, 5.0 and 20 mM for 48 hours. At the time of harvest, cell count and viability were measured by 0.4% trypan blue exclusion. SeeFigures 1A-1D Compounds 134 and 473 had the least effect on cell growth rate and viability remained >90% over the widest range of drug concentrations (see Figures 13 and 14, respectively). Figure 1B and 1C ).

[0952] Western blot analysis was also performed to determine the level of HbF induction by compounds 73, 134, 473, and 236 in KU812 cells. Hydroxyurea and hemin were used as positive controls, and β-actin was used as an internal protein control. Compounds 73, 134, and 473 all showed HbF induction, and compounds 134 and 473 increased HbF Figure 1E ) at concentrations between 0.5 and 5 μΜ. Compound 236 did not show significant HbF induction at the concentrations tested by Western blot analysis.

[0953] Flow cytometry was performed on KU812 cells after treatment with different concentrations (0.5, 2.5, 5, 10, and 20 μΜ) of compounds 473 and 236. An increase in the number of HbF positive cells (F-cells) and an increase in the mean fluorescence intensity (MFI) were observed for compound 473 Figure 2 ). In contrast, compound 236 did not significantly increase F-cells at these concentrations, but an increase in MFI was observed (data not shown). Compounds 73 and 134 were not analyzed by flow cytometry.

[0954] Example 2

[0955] To test the compounds under conditions that more closely mimic physiological conditions in a patient with SCD, sickle erythroid progenitor cells were cultured for 10 days and then treated with compound 473 at concentrations of 0.5 μΜ and 2.5 μΜ for 48 hours. The expression levels of HbF, HbS, and β-actin were analyzed by Western blot for the treated cells relative to cells treated with DMSO, hemin, or HU. The same treated cells were also analyzed by flow cytometry for γ-globin gene expression relative to cells treated with DMSO, hemin, or HU. Compound 473 (0.5 μΜ and 2.5 μΜ) induced γ-globin gene expression 1.6-fold and 1.9-fold, respectively, without affecting HbS protein levels. See Figures 15 and 16, respectively. Figure 3A An increase in F-cell levels was observed by flow cytometry. See Figure 17. Figure 3B .

[0956] Antisickling activity was observed in treated cells under hypoxic conditions. As described above, sickle red progenitor cells were cultured for 10 days and then treated with Compound 473 at concentrations of 0.5 μΜ and 2.5 μΜ or with hemin (about 50 μΜ) or with HU (about 100 μΜ) for 48 hours. The treated cells were then placed under hypoxic conditions (1% 02 and 5% C02). Cells treated with Compound 473 at concentrations of 0.5 μΜ and 2.5 μΜ significantly reduced the percentage of sickle cells compared to DMSO controls. See Figure 4A and Figure 4B .

Claims

1. Use of a compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating sickle cell disease in a subject, wherein said compound is: 1-Methyl-2-(6-trifluoromethyl-benzothiazol-2-ylamino)-1H-benzimidazole-5-carboxylic acid dimethylcarbamoylmethyl-amide, or 1-Methyl-2-(6-trifluoromethyl-benzothiazol-2-ylamino)-1H-benzimidazole-5-carboxylic acid [2-(2-hydroxy-ethoxy)-ethyl]-amide, or N-[2-(2-hydroxyethoxy)ethyl]-3-methyl-2-[[6-(trifluoromethyl)-1,3-benzothiazolyl-2-yl]amino]imidazo[4,5-b]pyridine-6-carboxamide Or its pharmaceutically acceptable salt.

2. The use of claim 1, wherein the compound is 1-methyl-2-(6-trifluoromethyl-benzothiazol-2-ylamino)-1H-benzimidazole-5-carboxylic acid dimethylcarbamoylmethyl-amide or a pharmaceutically acceptable salt thereof.

3. The use of claim 1, wherein the compound is 1-methyl-2-(6-trifluoromethyl-benzothiazol-2-ylamino)-1H-benzimidazole-5-carboxylic acid [2-(2-hydroxy-ethoxy)-ethyl]-amide or a pharmaceutically acceptable salt thereof.

4. The use of claim 1, wherein the compound is N-[2-(2-hydroxyethoxy)ethyl]-3-methyl-2-[[6-(trifluoromethyl)-1,3-benzothiazolyl-2-yl]amino]imidazo[4,5-b]pyridine-6-carboxamide or a pharmaceutically acceptable salt thereof.

5. The use as claimed in claim 1, wherein The intended use increases the expression of fetal hemoglobin (HbF) in the subjects.

6. The use as claimed in claim 1, wherein The intended use is to inhibit HbS polymerization or reduce reactive oxygen species (ROS) levels in the subjects, or to increase dissolved oxygen levels in the subjects' blood.

7. The use as claimed in claim 1, wherein The intended use is to reduce blood cell sickling in response to decreased air pressure, decrease atmospheric pressure, decrease oxygen partial pressure, or reduce hypoxia in the subjects.

8. The use as claimed in claim 1, wherein The intended use: Reduce the incidence or probability of pain crises, reduce the incidence or probability of pain crises requiring hospitalization, reduce the incidence of chest syndromes, reduce the number of transfusion events, or reduce the number of transfusion units per event.

9. The use as claimed in claim 1, wherein The stated purpose is to treat hemolytic anemia, vascular occlusive crisis, or multi-organ damage caused by microinfarction.

10. The use of claim 1, wherein the subject exhibits one or more clinical symptoms of sickle cell disease, β-thalassemia, or a related condition caused by said sickle cell disease.

11. The use of claim 1, wherein the subject exhibits genetic or biochemical markers of sickle cell disease, β-thalassemia, or a related condition caused by said sickle cell disease.

12. The use as claimed in claim 1, wherein the subject has sickle cell disease, β-thalassemia, or a condition associated with sickle cell disease.

13. The use as claimed in claim 1, wherein the medicament comprises another active compound selected from the group consisting of free hydroxyurea, dimethyl fumarate, monomethyl fumarate and methylbardoxolone.

14. The use as claimed in claim 1, wherein the subject is administered the compound at a dose of 10 mg / day to 100 mg / day.

15. Use of a pharmaceutical composition in the preparation of a medicament for treating sickle cell disease in a subject, wherein the pharmaceutical composition comprises a compound and a pharmaceutically acceptable carrier, said compound being: 1-Methyl-2-(6-trifluoromethyl-benzothiazol-2-ylamino)-1H-benzimidazole-5-carboxylic acid dimethylcarbamoylmethyl-amide, or 1-Methyl-2-(6-trifluoromethyl-benzothiazol-2-ylamino)-1H-benzimidazole-5-carboxylic acid [2-(2-hydroxy-ethoxy)-ethyl]-amide, or N-[2-(2-hydroxyethoxy)ethyl]-3-methyl-2-[[6-(trifluoromethyl)-1,3-benzothiazolyl-2-yl]amino]imidazo[4,5-b]pyridine-6-carboxamide Or its pharmaceutically acceptable salt.

16. The use as claimed in claim 15, wherein the drug is formulated into capsules or tablets.

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