Compositions and methods for treating systemic lupus erythematosus

Abstract

The present invention discloses a method for treating systemic lupus erythematosus using a compound or pharmaceutical composition that modulates the activity of a Bcl-2 family protein. In some methods, the patient to be treated is diagnosed with lupus nephritis. In some methods, the compound or pharmaceutical composition is administered to a patient in need thereof at a dose sufficient to cause one or more effects selected from the following: a decrease in protein excretion in the patient's urine, a decrease in the patient's serum anti-dsDNA autoantibody level, a decrease in the severity of the patient's skin lesions, a decrease in the severity of the patient's lymphadenopathy, a decrease in the severity of the patient's glomerulonephritis, a decrease in the severity of the patient's vasculitis, a decrease in the number of lymphocytes in a peripheral blood mononuclear cell (PMBC) population collected from the patient, a decrease in the number of lymphocytes in the patient's spleen, and a decrease in lymphocytic infiltration in the patient's kidney.

Description

[Technical Field]

[0001] This disclosure relates to a method for treating systemic lupus erythematosus (SLE) using a Bcl-2 inhibitor compound or a composition containing the same. [Background technology]

[0002] Apoptosis, or programmed cell death, is a fundamental biological process of tissue homeostasis. In mammals, it has been shown to regulate early embryonic development. In later stages of life, cell death is the default mechanism, eliminating potentially dangerous cells (e.g., cells with cancerous defects). Several apoptotic pathways are known. One of the most important apoptotic pathways concerns the Bcl-2 protein family, key regulators of the mitochondrial (also called "endogenous") pathway of apoptosis. See Daniel and Korsmeyer, Cell 116:205-219 (2004). Structural homologous domains BH1, BH2, BH3, and BH4 are characteristic of the Bcl-2 family proteins. The Bcl-2 protein family can be further classified into three subfamilies, specifically depending on how many homologous domains each protein contains and its biological activity, i.e., whether it has pro-apoptotic or anti-apoptotic function.

[0003] The first subgroup of Bcl-2 proteins includes proteins that possess all four homologous structural domains, namely BH1, BH2, BH3, and BH4. Their general effect is anti-apoptotic, i.e., protecting cells from the initiation of the cell death process. Proteins such as Bcl-2, Bcl-w, Bcl-xL, Mcl-1, and Bfl-l / Al are members of this first subgroup. Proteins belonging to the second subgroup of Bcl-2 proteins contain three homologous structural domains, BH1, BH2, and BH3, and have an apoptotic effect. Two main representative proteins of the second subgroup are Bax and Bak. The third subgroup of Bcl-2 proteins consists of proteins that contain only the BH3 structural domain, and members of this subgroup are usually called "BH3-only proteins." Their biological effect on cells is apoptosis promotion. Bim, Bid, Bad, Bik, Noxa, Hrk, Bmf, and Puma are examples of this third protein subfamily. The precise mechanism by which Bcl-2 family proteins regulate cell death is not yet fully understood. One hypothesis regarding the regulation of cell death by Bcl-2 family proteins suggests that BH3-only proteins are further classified into "activators," e.g., Bim and Bid, or "sensitizers," e.g., Bad, Bik, Noxa, Hrk, Bmf and Puma, with the choice of protein determined by its regulatory function.

[0004] One key to tissue homeostasis is achieving a balance of interactions among the three subgroups of cellular Bcl-2 proteins. Research has elucidated the mechanism by which the pro-apoptotic and anti-apoptotic subgroups of the Bcl-2 family proteins interact to enable programmed cell death. In cells, post-translational or transcriptional activation of BH3-only proteins occurs after receiving intracellular or extracellular signals. BH3-only proteins are the main inducers of the apoptosis cascade, one of which involves the activation of the pro-apoptotic proteins Bax and Bak in the cellular mitochondrial membrane. After activating Bax and / or Bak, which are already attached to or have migrated to the mitochondrial membrane, Bax and / or Bak oligomerize, causing mitochondrial outer membrane translocation (MOMP), cytochrome C release, and downstream activation of effector caspases, ultimately leading to apoptosis. According to the hypotheses of some researchers, some BH3-only proteins, such as Puma, Bim, and Bid, are "activators" because they directly bind to the pro-apoptotic proteins Bax and Bak and initiate MOMP, while other BH3-only proteins, such as Bad, Bik, and Noxa, are "sensitizers" that bind to anti-apoptotic proteins (e.g., Bcl-2, Bcl-xL, Bcl-w, Mcl-1), displacing and "releasing" the "activator" BH3-only proteins, and then indirectly inducing cell death by binding to and activating the pro-apoptotic proteins, such as Bax and Bak, and inducing oligomerization of Bax and Bak. Other studies suggest that anti-apoptotic proteins directly bind to and sequester Bax and Bak, while all BH3-only proteins modulate this interaction by binding to anti-apoptotic proteins, such as Bcl-2, Bcl-xL, Bcl-w, Mcl-1, which causes the release of Bax and Bak. See Adams and Cory, Oncogene 26:1324-1337 (2007) and Willis et al., Science 315:856-859 (2007).While the precise interaction between anti-apoptotic and pro-apoptotic Bcl-2 family proteins and their regulation of apoptosis is still under investigation, there is considerable scientific evidence suggesting that compounds that inhibit the binding of BH3-only proteins to anti-apoptotic Bcl-2 family proteins promote apoptosis.

[0005] Dysregulated apoptotic pathways are relevant to the pathology of numerous important diseases, such as neurodegenerative diseases like Alzheimer's disease (upward-regulated apoptosis) and proliferative diseases like cancer, autoimmune diseases, and prothrombotic conditions (downward-regulated apoptosis).

[0006] In particular, Bcl-2 may be an attractive therapeutic target for treating autoimmune disorders, including systemic lupus erythematosus (SLE). For example, dysregulation of apoptosis in patients diagnosed with SLE due to inadequate Bcl-2 function can lead to SLE-related complications, including lupus nephritis, by causing the survival of autoreactive immune cells, which is the most common sign of late SLE. Several studies have shown that Bcl-2 expression levels on renal infiltrating B and T cells are abnormally elevated compared to activated lymphocytes from healthy lymphoid tissue. See, for example, Ko et al., Arthritis & Rheumatology 68:2740-2751 (2016).

[0007] To treat proliferative disorders such as autoimmune diseases including systemic lupus erythematosus, there is always a need for small molecules that selectively inhibit the activity of certain types or parts of the Bcl-2 protein. [Overview of the Initiative]

[0008] This specification discloses a method for treating systemic lupus erythematosus in a patient, the method comprising administering a compound of formula (I) to a patient in need thereof. [ka] comprising administering a therapeutically effective amount thereof, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer or tautomer thereof, wherein A, E and Y are as defined herein.

[0009] The present specification further discloses a method of treating systemic lupus erythematosus in a patient, the method comprising administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising the compound of formula (I) as described above, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer or tautomer thereof.

[0010] In one embodiment, the method of treating systemic lupus erythematosus in a patient comprises administering to a patient in need thereof a compound of formula (I),

Chemical

Chemical

[0011] In one embodiment, the method further provides to a patient in need a compound of formula (I) given by formula (II), [ka] Or, including administering a therapeutically effective dose of a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, or tautomer thereof.

[0012] In one embodiment, the method provides to a patient who needs it. [ka] , Or, administering a therapeutically effective amount of a compound selected from the group consisting of pharmaceutically acceptable salts, solvates, hydrates, or tautomers thereof.

[0013] In one embodiment, the method comprises administering a therapeutically effective amount of a compound to a patient in need thereof, wherein the compound is [ka] , or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof.

[0014] In one embodiment, the method comprises administering a therapeutically effective amount of a compound to a patient in need thereof, wherein the compound is [ka] , or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof.

[0015] In one embodiment, the method involves administering to a patient in need a therapeutically effective amount of a compound selected from the group of compounds listed in Table 1, as described herein.

[0016] In one embodiment, a method for treating systemic lupus erythematosus in a patient involves providing a patient who requires it with a drug composition containing compound (I). [ka] Or, administering a therapeutically effective dose of a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, or tautomer thereof, wherein A is [ka] Selected from the group consisting of, E is TIFF0007914089000013.tif35 is a carbon atom with a double bond, TIFF0007914089000014.tif35 has a single bond -C(H)-, and TIFF0007914089000015.tif35 is selected from the group consisting of nitrogen atoms with single bonds, Y is selected from -C(H)- and -O-, R 1 is hydrogen and -N(R 7a )(R 7b ) are selected from, R 2 , R 3 , R 4 , R 5 and R 6 These are, independently, hydrogen and optionally substituted C. 1-6 Alkyl, optionally substituted C 3-6Selected from the group consisting of cycloalkyl, heterocyclo, optionally substituted heteroaryl, and (heterocyclo)alkyl, R 7a is an arbitrarily substituted C 1-6 Selected from alkyl and optionally substituted (heterocyclo)alkyl, R 7b is hydrogen and C 1-4 Selected from alkyl groups.

[0017] In one embodiment, the method provides a patient in need with a drug composition further comprising a compound of formula (I) given by formula (II), [ka] Or, including administering a therapeutically effective dose of a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, or tautomer thereof.

[0018] In one embodiment, the method comprises administering a therapeutically effective amount of a drug composition to a patient in need thereof, wherein the drug composition is [ka] , or a compound selected from the group consisting of pharmaceutically acceptable salts, solvates, hydrates, or tautomers thereof.

[0019] In one embodiment, the method comprises administering a therapeutically effective amount of a drug composition to a patient in need thereof, wherein the drug composition comprises the following compounds: [ka] or comprising pharmaceutically acceptable salts, solvates, hydrates, or tautomers thereof.

[0020] In one embodiment, the method comprises administering a therapeutically effective amount of a drug composition to a patient in need thereof, wherein the drug composition is [ka] or a compound selected from the group consisting of pharmaceutically acceptable salts, solvates, hydrates, or tautomers thereof.

[0021] In one embodiment, the method involves administering to a patient in need a therapeutically effective amount of a drug composition comprising a compound selected from the group of compounds listed in Table 1, as described herein.

[0022] In one embodiment, a method for treating a patient's systemic lupus erythematosus comprises administering a therapeutically effective amount of a compound to a patient in need, wherein the compound is (S)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide.

[0023] In one embodiment, the patient requiring the treatment is diagnosed with lupus nephritis.

[0024] In one embodiment, the compound or drug composition is administered to a patient in need in a dose sufficient to produce one or more effects selected from the group consisting of: a decrease in the patient's urinary protein excretion; a decrease in the patient's serum anti-dsDNA autoantibody levels; a decrease in the patient's skin injury severity; a decrease in the patient's lymphadenopathy severity; a decrease in the patient's glomerulonephritis severity; a decrease in the patient's vasculitis severity; a decrease in lymphocyte count in peripheral blood mononuclear cells (PMBCs) collected from the patient; a decrease in lymphocyte count in the patient's spleen; and a decrease in renal lymphocyte infiltration.

[0025] In one embodiment, the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the patient's urinary protein excretion.

[0026] In one example, the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the patient's serum anti-dsDNA autoantibody levels.

[0027] In one embodiment, the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the severity of the patient's skin injury.

[0028] In one embodiment, the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the severity of the patient's lymph node swelling.

[0029] In one embodiment, the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the severity of the patient's glomerulonephritis.

[0030] In one embodiment, the compound or drug composition is used to provide the patient who needs it. interstitial It is administered in a dose sufficient to reduce the severity of nephritis.

[0031] In one embodiment, the compound or drug is administered to a patient in need in a dose sufficient to reduce the severity of the patient's vasculitis.

[0032] In one embodiment, the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the lymphocyte count in peripheral blood mononuclear cells (PMBCs) collected from the patient.

[0033] In one embodiment, the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the number of lymphocytes in the patient's spleen.

[0034] In one embodiment, the compound drug composition is administered to a patient in need in a dose sufficient to reduce lymphocyte infiltration in the patient's kidneys.

[0035] In one example, compound (I), [ka] Or its pharmaceutically acceptable salts, solvates, hydrates, stereoisomers, or tautomers, for use in the preparation of drugs for the treatment of systemic lupus erythematosus in patients in need, here, A is [ka] Selected from the group consisting of, E is TIFF0007914089000022.tif35 is a carbon atom with a double bond, TIFF0007914089000023.tif35 has a single bond -C(H)-, and TIFF0007914089000024.tif35 is selected from the group consisting of nitrogen atoms with single bonds, Y is selected from -C(H)- and -O-, R 1 is hydrogen and -N(R 7a )(R 7b ) are selected from, R 2 , R 3 , R 4 , R 5 and R 6 These are, independently, hydrogen and optionally substituted C. 1-6 Alkyl, optionally substituted C 3-6 Selected from the group consisting of cycloalkyl, heterocyclo, optionally substituted heteroaryl, and (heterocyclo)alkyl, R 7a is an arbitrarily substituted C 1-6 Selected from alkyl and optionally substituted (heterocyclo)alkyl, R 7b is hydrogen and C 1-4 Selected from alkyl groups.

[0036] In one example, the compound used for the preparation of the drug is further a compound of formula (I) given by formula (II), [ka] Or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, or tautomer thereof.

[0037] In one example, the compound used for the preparation of the drug is: [ka] , Alternatively, a selection from the group consisting of pharmaceutically acceptable salts, solvates, hydrates, or tautomers thereof.

[0038] In one example, the compound used for the preparation of the drug is: [ka] or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof.

[0039] In one example, the compound used for the preparation of the drug is: [ka] or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof.

[0040] In one example, the compound used for the preparation of the drug is a compound selected from the group of compounds listed in Table 1, as described herein.

[0041] In one example, (S)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide is intended to be a drug for the treatment of systemic lupus erythematosus in patients who require it.

[0042] In one embodiment, the drug is used to treat systemic lupus erythematosus in a patient diagnosed with lupus nephritis.

[0043] In one embodiment, the compound or drug composition is for the production of a drug for the treatment of systemic lupus erythematosus, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to produce one or more effects selected from the group consisting of: a decrease in the patient's urinary protein excretion; a decrease in the patient's serum anti-dsDNA autoantibody levels; a decrease in the patient's skin injury severity; a decrease in the patient's lymphadenopathy severity; a decrease in the patient's glomerulonephritis severity; a decrease in the patient's vasculitis severity; a decrease in the lymphocyte count in the peripheral blood mononuclear cell (PMBC) group taken from the patient; a decrease in the lymphocyte count in the patient's spleen; and a decrease in the patient's renal lymphocyte infiltration.

[0044] In one embodiment, the compound or drug composition is for the purpose of producing a drug for the treatment of systemic lupus erythematosus, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the patient's urinary protein excretion.

[0045] In one embodiment, the compound or drug composition is for the purpose of producing a drug for the treatment of systemic lupus erythematosus, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the patient's serum anti-dsDNA autoantibody levels.

[0046] In one embodiment, the compound or drug composition is for the purpose of producing a drug for the treatment of systemic lupus erythematosus, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the severity of the patient's skin damage.

[0047] In one embodiment, the compound or drug composition is for the purpose of producing a drug for the treatment of systemic lupus erythematosus, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the severity of the patient's lymphadenopathy.

[0048] In one embodiment, the compound or drug composition is for the purpose of producing a drug for the treatment of systemic lupus erythematosus, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the severity of the patient's glomerulonephritis.

[0049] In one embodiment, the compound or drug composition is for the purpose of producing a drug for the treatment of systemic lupus erythematosus, wherein the compound or drug composition is provided to a patient in need of the patient interstitial It is administered in a dose sufficient to reduce the severity of nephritis.

[0050] In one embodiment, the compound or drug composition is for the purpose of producing a drug for the treatment of systemic lupus erythematosus, wherein the compound or drug is administered to a patient in need in a dose sufficient to reduce the severity of the patient's vasculitis.

[0051] In one embodiment, the compound or drug composition is for the purpose of producing a drug for the treatment of systemic lupus erythematosus, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the lymphocyte count in peripheral blood mononuclear cells (PMBCs) collected from the patient.

[0052] In one embodiment, the compound or drug composition is for the purpose of producing a drug for the treatment of systemic lupus erythematosus, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the number of lymphocytes in the patient's spleen.

[0053] In one embodiment, the compound or drug composition is for the purpose of producing a drug for the treatment of systemic lupus erythematosus, wherein the compound drug composition is administered to a patient in need in a dose sufficient to reduce lymphocyte infiltration in the patient's kidneys.

[0054] In one example, a compound for treating systemic lupus erythematosus in a patient requiring it is compound (I), [ka] or comprising a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or tautomer thereof, wherein, A is [ka] Selected from the group consisting of, E is TIFF0007914089000031.tif35 is a carbon atom with a double bond, TIFF0007914089000032.tif35 has a single bond -C(H)-, and TIFF0007914089000033.tif35 is selected from the group consisting of nitrogen atoms with single bonds, Y is selected from -C(H)- and -O-, R 1 is hydrogen and -N(R 7a )(R 7b ) are selected from, R 2 , R 3 , R 4 , R 5 and R 6 These are, independently, hydrogen and optionally substituted C. 1-6Alkyl, optionally substituted C 3-6 Selected from the group consisting of cycloalkyl, heterocyclo, optionally substituted heteroaryl, and (heterocyclo)alkyl, R 7a is an arbitrarily substituted C 1-6 Selected from alkyl and optionally substituted (heterocyclo)alkyl, R 7b is hydrogen and C 1-4 Selected from alkyl groups.

[0055] In one example, a compound for treating systemic lupus erythematosus in a patient requiring it is further a compound of formula (I) given by formula (II), [ka] or including pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, or tautomers thereof.

[0056] In one example, a compound for treating systemic lupus erythematosus in patients who require it is: [ka] , or a compound selected from its pharmaceutically acceptable salts, solvates, hydrates, or tautomers.

[0057] In one example, a compound for treating systemic lupus erythematosus in patients who require it is: [ka] or comprising pharmaceutically acceptable salts, hydrates, solvates, or tautomers thereof.

[0058] In one example, a compound for treating systemic lupus erythematosus in patients who require it is: [ka] or comprising pharmaceutically acceptable salts, hydrates, solvates, or tautomers thereof.

[0059] In one example, the compound for treating systemic lupus erythematosus in a patient requiring it includes a compound selected from the group of compounds listed in Table 1, as described herein.

[0060] In one example, the compound for treating systemic lupus erythematosus in patients requiring it comprises (S)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide.

[0061] In one embodiment, the compound is used to treat systemic lupus erythematosus in patients diagnosed with lupus nephritis.

[0062] In one embodiment, the compound is for the treatment of systemic lupus erythematosus in a patient, wherein the compound or drug composition is administered to the patient in need in a dose sufficient to produce one or more effects selected from the group consisting of: a decrease in the patient's urinary protein excretion; a decrease in the patient's serum anti-dsDNA autoantibody levels; a decrease in the patient's skin injury severity; a decrease in the patient's lymphadenopathy severity; a decrease in the patient's glomerulonephritis severity; a decrease in the patient's vasculitis severity; a decrease in lymphocyte count in peripheral blood mononuclear cells (PMBCs) collected from the patient; a decrease in lymphocyte count in the patient's spleen; and a decrease in renal lymphocyte infiltration.

[0063] In one embodiment, the compound is used to treat systemic lupus erythematosus in a patient, where the compound or drug composition is administered to the patient in need in a dose sufficient to reduce the patient's urinary protein excretion.

[0064] In one embodiment, the compound is used to treat systemic lupus erythematosus in a patient, where the compound or drug composition is administered to the patient in need in a dose sufficient to reduce the patient's serum anti-dsDNA autoantibody levels.

[0065] In one embodiment, the compound is used to treat systemic lupus erythematosus in a patient, where the compound or drug composition is administered to the patient in need in a dose sufficient to reduce the severity of the patient's skin damage.

[0066] In one embodiment, the compound is used to treat systemic lupus erythematosus in a patient, where the compound or drug composition is administered to the patient in need in a dose sufficient to reduce the severity of the patient's lymphadenopathy.

[0067] In one embodiment, the compound is used to treat systemic lupus erythematosus in a patient, where the compound or drug composition is administered to the patient in need in a dose sufficient to reduce the severity of the patient's glomerulonephritis.

[0068] In one embodiment, the compound is for the treatment of systemic lupus erythematosus in a patient, and the compound or drug composition is used to treat a patient in need of it. interstitial It is administered in a dose sufficient to reduce the severity of nephritis.

[0069] In one embodiment, the compound is used to treat systemic lupus erythematosus in a patient, where the compound or drug is administered to the patient in need in a dose sufficient to reduce the severity of the patient's vasculitis.

[0070] In one embodiment, the compound is used to treat systemic lupus erythematosus in a patient, where the compound or drug composition is administered to the patient in need in a dose sufficient to reduce the lymphocyte count in peripheral blood mononuclear cells (PMBCs) collected from the patient.

[0071] In one embodiment, the compound is for the treatment of systemic lupus erythematosus in a patient, where the compound or drug composition is administered to the patient in need in a dose sufficient to reduce the number of lymphocytes in the patient's spleen.

[0072] In one embodiment, the compound is used to treat systemic lupus erythematosus in a patient, where the compound drug composition is administered to the patient in need in a dose sufficient to reduce lymphocyte infiltration in the patient's kidneys. [Brief explanation of the drawing]

[0073] [Figure 1] This line graph shows the time course of protein concentration in the urine of MRL / lpr mice treated with the vehicle or compound 1. [Figure 2] This bar graph shows serum anti-dsDNA autoantibody levels in MRL / lpr mice treated with the vehicle or compound 1. [Figure 3] This line graph shows the time course of skin injury scores in MRL / lpr mice treated with the vehicle or compound 1. [Figure 4] This line graph shows the time course of lymph node scores in MRL / lpr mice treated with the vehicle or compound 1. [Figure 5] This is a series of bar graphs showing pathological scores for glomerulonephritis, interstitial nephritis, vasculitis, and total lupus nephritis in MRL / lpr mice treated with the vehicle or compound 1. [Figure 6] This bar graph shows the lymphocyte count in peripheral blood mononuclear cells (PMBCs) collected from MRL / lpr mice treated with the vehicle or compound 1. [Figure 7] This bar graph shows the lymphocyte count in the spleen of MRL / lpr mice treated with the vehicle or compound 1. [Figure 8] This is a series of figures showing the lymphocyte count in the kidneys of MRL / lpr mice treated with the vehicle or compound 1. [Modes for carrying out the invention]

[0074] This disclosure relates to compounds that can modulate Bcl-2 family proteins, or pharmaceutically acceptable salts, solvates, hydrates, tautomers and stereoisomers thereof. Compounds that can modulate Bcl-2 family proteins can be used to treat, prevent or improve diseases and disorders associated with the activity of Bcl-2 family proteins.

[0075] In some embodiments, the Disclosure describes a method for treating or improving systemic lupus erythematosus (SLE) in a patient by administering to the patient in need a therapeutically effective dose of a compound capable of modulating Bcl-2 family proteins, or a drug composition comprising a compound capable of modulating Bcl-2 family proteins.

[0076] definition In this disclosure, the term "alkyl" means, when used by itself or as part of another group, 1 to 12 carbon atoms, i.e., C 1-12 Alkyl, or a specified number of carbon atoms, for example, C1 alkyl like methyl, C2 alkyl like ethyl, C3 alkyl like propyl or isopropyl, C like methyl, ethyl, propyl or isopropyl 1-3 This refers to unsubstituted linear or branched aliphatic hydrocarbon compounds, including alkyl groups. In one example, the alkyl group is a linear C 1-6 It is an alkyl group. In another example, the alkyl group is a branched chain C 3-6 It is an alkyl group. In another example, the alkyl group is a linear C 1-4 It is an alkyl group. In another example, the alkyl group is a branched chain C 3-4It is an alkyl group. In another example, the alkyl group is a linear or branched C chain. 3-4 It is an alkyl group. In another embodiment, the alkyl group is partially or completely deuterated, i.e., one or more hydrogen atoms of the alkyl group are replaced by deuterium atoms. Non-limiting exemplary C 1-12 Examples of alkyl groups include methyl, -CD3, ethyl, propyl, isopropyl, butyl, s-butyl, t-butyl, isobutyl, 3-pentyl, hexyl, heptyl, octyl, nonyl, and decyl. (Non-limiting, exemplary C) 1-4 Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, s-butyl, t-butyl, and isobutyl. (Non-limiting exemplary C) 1-4 Examples of the groups include methyl, ethyl, propyl, isopropyl, and t-butyl.

[0077] In this disclosure, the term "optionally substituted alkyl" means, when used by itself or as part of another group, an alkyl group that is unsubstituted or substituted with one, two, or three substituents, these substituents independently selected from the group consisting of halo, nitro, cyano, hydroxy, alkoxy, amino, alkylamino, dialkylamino and optionally substituted aryl groups. In one example, the optionally substituted alkyl group is substituted with two substituents. In another example, the optionally substituted alkyl group is substituted with one substituent. In yet another example, the optionally substituted alkyl group is unsubstituted. Non-limiting exemplary optionally substituted alkyl groups include -CH2Ph, -CH2CH2NO2, -CH2CH2OH, -CH2CH2OCH3 and -CH2CH2F.

[0078] In this disclosure, the term "cycloalkyl," when used by itself or as part of another group, refers to an unsubstituted saturated or partially unsaturated (e.g., containing one or two double bonds) cyclic aliphatic hydrocarbon, which contains 1 to 3 rings and has 3 to 12 carbon atoms, i.e., C 3-12A cycloalkyl group, or a specified number of carbon atoms. In one example, the cycloalkyl group has two rings. In another example, the cycloalkyl group has one ring. In yet another example, the cycloalkyl group has C 3-8 It is a cycloalkyl group. In another example, the cycloalkyl group is C 3-6 It is a cycloalkyl group. In another example, the cycloalkyl group is C 3-5 They are cycloalkyl compounds. Non-limiting exemplary cycloalkyl compounds include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, norbornyl, decahydronaphthalene, adamantyl, cyclohexenyl, cyclopentenyl, cyclopentanone, spiro[3.3]heptane, and bicyclo[3.3.1]nonane.

[0079] In this disclosure, the term “optionally substituted cycloalkyl” means, when used by itself or as part of another group, an unsubstituted cycloalkyl group or a cycloalkyl group substituted with one, two, or three substituents, the substituents independently selected from the group consisting of halo, nitro, cyano, hydroxy, alkyl, alkoxy, amino, alkylamino, dialkylamino, haloalkyl, and heterocyclo. In one example, the optionally substituted cycloalkyl group is substituted with two substituents. In another example, the optionally substituted cycloalkyl group is substituted with one substituent. In yet another example, the optionally substituted cycloalkyl group is unsubstituted.

[0080] In this disclosure, the term “haloalkyl” refers to an alkyl group substituted with one or more fluorine, chlorine, bromine, and / or iodine atoms, either by itself or as part of another group. In one example, the alkyl group is substituted with one, two, or three iodine and / or chlorine atoms. In another example, the haloalkyl group is C 1-4These are haloalkyl groups. Non-limiting exemplary haloalkyl groups include fluoromethyl, 2-fluoroethyl, difluoromethyl, trifluoromethyl, pentafluoroethyl, 1,1-difluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, 3,3,3-trifluoropropyl, 4,4,4-trifluorobutyl, and trichloromethyl.

[0081] In this disclosure, the term “alkoxy” refers, when used by itself or as part of another group, to an optionally substituted alkyl group bonded to a terminal oxygen atom. In one example, the alkoxy group is a C bonded to a terminal oxygen atom. 1-6 It is alkyl. In another example, the alkoxy group is a C bonded to the terminal oxygen atom. 1-4 It is alkyl. Non-limiting exemplary alkoxys include methoxy, ethoxy, and t-butoxy.

[0082] In this disclosure, the term “heterocyclo,” when used by itself or as part of another group, refers to an unsubstituted saturated and partially unsaturated cyclic group (e.g., containing one or two double bonds) which contains one, two or three rings and has 3 to 14 ring members, i.e., a 3 to 14-membered heterocyclo, where at least one carbon atom of one ring is replaced by a heteroatom. The term “heterocyclo” means to include cyclic ureido groups (e.g., imidazolidinyl-2-one), cyclic amide groups (e.g., β-lactam, γ-lactam, δ-lactam and ε-lactam), and cyclic urethane groups (e.g., oxazolidinyl-2-one). In one embodiment, the heterocyclo group is a 4, 5, 6, 7, or 8-membered cyclic group containing one ring and one or two oxygen and / or nitrogen atoms. In one embodiment, the heterocyclo group is a 5 or 6-membered cyclic group containing one ring and one or two nitrogen atoms. In one example, the heterocyclo group is an 8, 9, 10, 11, or 12-membered ring group containing two rings and one or two nitrogen atoms. In one example, the heterocyclo group is a 4 or 5-membered ring group containing one ring and one oxygen atom. Heterocyclos can selectively bond to other parts of a molecule via carbon or nitrogen atoms. Non-limiting exemplary heterocyclo groups include 1,4-dioxane, 2-oxopyrrolidine-3-yl, 2-imidazolidinone, piperidinyl, morpholinyl, piperazinyl, pyrrolidinyl, 8-azabicyclo[3.2.1]octane (nortropane), 6-azaspiro[2.5]octane, 6-azaspiro[3.4]octane, indolyl, indolyl-2-one, and 1,3-dihydro-2H-benzo[d]imidazole-2-one.

[0083] In this disclosure, the term “optionally substituted heterocyclo” as used herein means a heterocyclo that is unsubstituted, either by itself or as part of another group, or substituted with one, two, or three substituents, the substituents independently selected from the group consisting of halo, nitro, cyano, hydroxy, alkyl, alkoxy, amino, alkylamino, dialkylamino, haloalkyl and heterocyclo.

[0084] In this disclosure, the term "(heterocyclo)alkyl group" refers to an alkyl group substituted with one optionally substituted heterocyclo group, either by itself or as part of another group. In one example, the (heterocyclo)alkyl group is a C group substituted with one optionally substituted 4- to 6-membered heterocyclo group. 1-4 It is an alkyl group. Heterocyclo groups can be bonded to alkyl groups via carbon or nitrogen atoms. Non-limiting exemplary (heterocyclo)alkyl groups include: [ka]

[0085] As used herein, the term “heteroaryl” refers to a monocyclic aromatic group having 5 to 14 ring atoms, which comprises one or more ring heteroatoms selected from the group consisting of N, O, and S, with the remaining ring atom being C. Examples include, but are not limited to, furfuryl, thienyl, pyrrolyl, pyridyl, pyrazolyl, pyrimidinyl, imidazolyl, isoxazolyl, oxazolyl, oxadiazolyl, pyridinyl, thiophen-2-yl, isothiazole, thiazolyl, thiadiazole, triazolyl, and triazinyl.

[0086] As used herein, the term “optionally substituted heteroaryl” means a heteroaryl that is either unsubstituted or substituted with one, two, or three substituents, the substituents being independently selected from the group consisting of halo, nitro, cyano, hydroxy, alkyl, alkoxy, amino, alkylamino, dialkylamino, haloalkyl, and heterocyclo.

[0087] As used in this disclosure, the article “a / an” refers to one or more of the grammatical objects of the article. For example, “one element” refers to one or more of the elements.

[0088] Unless otherwise specified, the term "and / or" as used in this disclosure means "and" or "or".

[0089] As used herein, the term “about” includes ±10% of the aforementioned number. Therefore, “about 10” means between 9 and 11.

[0090] As used herein, the term “isomer” refers to compounds having the same composition and molecular weight but different physical and / or chemical properties. Structural differences may be in composition (e.g., geometric isomers) or in the ability to rotate the plane of polarization (stereoisomers). With respect to stereoisomers, the compounds of this disclosure may have one or more chiral carbon atoms and may appear as racemates, racemic mixtures, or as a single enantiomer or non-enantiomer.

[0091] As used herein, the term “stereoisomer” or “stereoisomeric form” is a general term for all isomers of a single molecule that differ only in the spatial orientation of their atoms. It includes enantiomers and isomers of compounds that are not mirror images of each other and have one or more chiral centers (an enantiomers).

[0092] The term "chiral center" or "chiral carbon atom" refers to a carbon atom bonded to four different groups.

[0093] The terms "enantiomer" and "of an enantiomer" refer to molecules that cannot be superimposed onto their mirror image, and therefore possess optical activity. Here, enantiomers rotate the plane of polarization in one direction, while their mirror image compounds rotate the plane of polarization in the opposite direction.

[0094] The term "racemic" refers to a mixture of equal amounts of enantiomers that are not optically active.

[0095] The term "absolute configuration" refers to the spatial arrangement of chiral molecular entities (or groups) and their stereochemical description, such as R or S.

[0096] Unless otherwise specified, the stereochemical terms and conventions used in this invention are intended to be consistent with those set forth in Pure & Appl. Chem 68:2193 (1996).

[0097] The term "enantiomer excess" or "ee" refers to a measure of how much of one enantiomer is present compared to another. For a mixture of R and S enantiomers, the enantiomer excess is defined as |RS|*100, where R and S are the mole or weight fractions of the enantiomers in the mixture, respectively, such that R+S=1. By knowing the optical rotation of a chiral substance, the enantiomer excess can be calculated as ([α] obs / [α] max )*100 is defined as, where [α] obs [α] is the optical rotation of the enantiomer mixture. max This represents the optical rotation of the pure enantiomeric isomers. The enantiomeric excess can be determined using various analytical techniques (including NMR spectroscopy, chiral column chromatography, or optical polarization measurement).

[0098] The terms "enantiomerically pure" or "enantiopurine" refer to a sample of a chiral substance in which all molecules (within the detection limit) have the same chiral meaning. In one example, a compound of the present disclosure having one or more chiral centers is enantiopurine.

[0099] The terms "enantiomerically enriched" or "enantiomerically enriched" refer to samples of chiral substances where the enantiomer excess exceeds 50%, for example, approximately 60%, 70%, 80%, 90%, 95%, 98%, or 99%. Enantiomerically enriched compounds may also be enantiomerically pure.

[0100] As used herein, the term “pharmaceutically acceptable salt” means any salt of the compounds herein that is physiologically acceptable in a target patient (e.g., a mammal such as human) (e.g., obtained by reaction with an acid or a base).

[0101] The terms "salt," etc., are used with the intention of also applying to salts of enantiomers, stereoisomers, rotational isomers, tautomers, positional isomers, and racemic mixtures of the compounds of the present invention.

[0102] As used herein, the term “solvate” refers to a combination, physical association and / or solvation of a compound of the Disclosure with a solvent molecule, e.g., a disolvate, monosolvate, or hemisolvate, where the ratio of the solvent molecule to the compound of the Disclosure is about 2:1, about 1:1, or about 1:2, respectively. Such physical associations involve varying degrees of ionic and covalent bonding, including hydrogen bonding. In some cases, solvates can be isolated, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. Thus, “solvate” encompasses solvates in solution and solvable solvates.

[0103] "Patient" or "subject" is a mammal such as a human, mouse, rat, guinea pig, dog, cat, horse, cattle, or pig, or a non-human primate such as a monkey, chimpanzee, baboon, or rhesus macaque.

[0104] As used herein, terms such as “treat,” “treating,” and “treatment” mean eliminating, reducing, or improving a disease or condition and / or symptoms associated therewith. Treatment of a disease or condition does not necessarily require the complete elimination of the disease or condition and / or symptoms associated therewith, although this is not excluded. The term “treatment” and its synonyms consider administering a therapeutically effective amount of the compounds of this disclosure to a subject requiring such treatment. Treatment may be carried out, depending on the symptoms, for example, to inhibit the symptoms. It may be achieved over a short period, over a medium period, or as long-term treatment, for example, in the background of maintenance therapy.

[0105] As used in this disclosure, the terms “administer,” “administering,” or “administration” refer to directly administering the disclosed compound, a pharmaceutically acceptable salt or composition of the disclosed compound to a subject, or directly administering a pharmaceutically acceptable salt or composition of the compound to a subject so that an equivalent amount of the active compound can be formed in vivo in the subject.

[0106] As used herein, the terms “prevent, preventing, and prevention” refer to methods of preventing the onset of a disease or condition and / or its associated symptoms, or preventing a subject from contracting the disease. As used herein, “prevent, preventing, and prevention” further include delaying the onset of a disease or condition and / or its associated symptoms, thereby reducing the risk of a subject contracting the disease. The terms “prevent, preventing, and prevention” may also include “preventive treatment,” and refer to reducing the likelihood of a disease or condition recurrence or a previously controlled disease or condition in a subject who is not currently suffering from the disease or condition or has not experienced a recurrence of such disease or condition, but is at risk of recurrence or relapse of such disease or condition, or is susceptible to recurrence or relapse of such disease or condition.

[0107] As used herein, the terms “therapeutic effective dose” or “effective dose” refer to an amount of one or more active ingredients sufficient to effectively deliver one or more active ingredients to an individual in need to treat a target condition or disease when administered by the method of this disclosure. In the case of proliferative disorders, such as autoimmune disorders, a therapeutic effective dose of the drug can reduce (i.e., delay, and preferably halt) undesirable cell proliferation.

[0108] As used in this disclosure, the term “carrier” encompasses carriers, excipients and diluents, and also means materials, compositions or vehicles that transport or deliver a drug agent from one organ or body part of a subject to another organ or body part of a subject, such as liquid or solid fillers, diluents, excipients, solvents or encapsulating materials.

[0109] In this disclosure, the terms “Bcl-2 protein” or “Bcl-2 protein family” refer to any one of the proteins Bax, Bak, Bid, Bcl-2, Bcl-xL, Mcl-1, Bcl-w, Bfl-l / Al, Bim, Puma, Bad, Bik / Blk, Noxa, Bmf, Hrk / DP5, and Beclin-1. See Cold Spring Harbor Perspect Biol 2013;5:a008714.

[0110] The terms “disease,” “pathological condition,” or “disorder” generally refer to a pathological condition or function, and mean a disorder and / or abnormality that may be expressed in the form of specific symptoms, symptoms, and / or dysfunctions. The compounds of this disclosure inhibit Bcl-2 proteins, e.g., Bcl-2 and / or Bcl-xL, and can be used for the treatment and prevention of diseases, pathological conditions, or disorders, e.g., hyperproliferative disorders, where inhibition of Bcl-2 proteins provides benefits.

[0111] The term "proliferative disorder" refers to any disorder in which a localized population of proliferating cells in an animal is not controlled by the normal limitations of proliferation. In one example, a proliferative disorder is an autoimmune disorder, such as systemic lupus erythematosus.

[0112] In some embodiments, the compounds of the present disclosure can be used to treat "Bcl-2 protein-mediated disorders," such as Bcl-2 mediated disorders and / or Bcl-xL mediated disorders. A Bcl-2 protein-mediated disorder is any pathological condition in which the Bcl-2 protein is known to play a role. In one embodiment, the Bcl-2 mediated disorder is a proliferative disorder. In one embodiment, the Bcl-2 mediated disorder is systemic lupus erythematosus.

[0113] Compounds of the Disclosure In one embodiment, this disclosure relates to a compound of formula (I): [ka] Or relating to pharmaceutically acceptable salts, hydrates, solvates, stereoisomers or tautomers thereof, here, A is [ka] Selected from the group consisting of, E is TIFF0007914089000041.tif35 is a carbon atom with a double bond, TIFF0007914089000042.tif35 has a single bond -C(H)-, and TIFF0007914089000043.tif35 is selected from the group consisting of nitrogen atoms with single bonds, Y is selected from -C(H)- and -O-, R 1 is hydrogen and -N(R 7a )(R 7b ) are selected from, R 2 , R 3 , R 4 , R 5 and R 6 These are, independently, hydrogen and optionally substituted C. 1-6 Alkyl, optionally substituted C 3-6 Selected from the group consisting of cycloalkyl, heterocyclo, optionally substituted heteroaryl, and (heterocyclo)alkyl, R 7a is selected from optionally substituted C 1-6 alkyl and optionally substituted (heterocyclo)alkyl, and R 7b is selected from hydrogen and C 1-4 alkyl.

[0114] In some embodiments, A is

Chemical Structure

Chemical Structure

Chemical Structure

Chemical Structure

Chemical Structure

Chemical Structure

[0115] In some embodiments, E is a carbon atom, wherein TIFF0007914089000050.tif35 is a double bond. In some embodiments, E is -C(H)-, wherein TIFF0007914089000051.tif35 is a single bond. In some embodiments, E is a nitrogen atom, wherein TIFF0007914089000052.tif35 is a single bond.

[0116] In some embodiments, R 1 is hydrogen. In some embodiments, R 1 is -N(R 7a )(R 7b ).

[0117] In some embodiments, R 2 is optionally substituted C 1-6 alkyl. In some embodiments, R 2 is optionally substituted C 1-4 alkyl. In some embodiments, R 2 is optionally substituted C3 alkyl. In some embodiments, R 2 is isopropyl. In some embodiments, R 2 is optionally substituted C 3-6 cycloalkyl. In some embodiments, R 2 is optionally substituted C 3-5 cycloalkyl. In some embodiments, R 2 is cyclopropyl. In some embodiments, R 2 is optionally substituted heteroaryl. In some embodiments, R 2 is pyridine.

[0118] In some embodiments, R 3 is (heterocyclo)alkyl. In some embodiments, R 3 is

Chemical Formula

[0119] In some embodiments, R 4 is (heterocyclo)alkyl. In some embodiments, R 4 is

Chemical Formula

[0120] In some examples, R 5 R is a (heterocyclo)alkyl. In some examples, R 5 teeth, [ka] In some examples, R 5 It is heterocycloidal. In some examples, R 5 It is tetrahydro-2H-pyranyl.

[0121] In some examples, R 6 R is a (heterocyclo)alkyl. In some examples, R 6 teeth, [ka] In some examples, R 6 It is heterocycloidal. In some examples, R 6 It is tetrahydro-2H-pyranyl.

[0122] In some examples, R 7a is an arbitrarily substituted C 1-6 It is alkyl. In some examples, R 7a is an arbitrarily substituted C 1-4 It is alkyl. In some examples, R 7a R is an optionally substituted methyl group. In some examples, R 7a R is methyl. In some examples, R 7a is an optionally substituted (heterocyclo)alkyl. In some examples, R 7a teeth, [ka] In some examples, R 7a teeth, [ka] , which is described. In some embodiments, R 7a is

Chemical Formula

[0123] In some embodiments, R 7b is hydrogen. In some embodiments, R 7b is C 1-4 alkyl. In some embodiments, R 7b is C 1-3 alkyl. In some embodiments, R 7b is methyl.

[0124] In some embodiments, the present disclosure relates to a compound of formula (I) selected from Table 1, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer or tautomer thereof.

[0125]

Table 1-1

Table 1-2

Table 1-3

Table 1-4

Table 1-5

[0126] Compounds of formula (I) may contain one or more chiral centers, thus generating enantiomers, non-enantiomers, and other stereoisomeric forms. This disclosure is intended to encompass the use of all such possible forms, including racemic and split forms, and mixtures thereof. In consideration of this disclosure, a single stereoisomer, for example, an enantiomer, can be isolated based on methods known in the art. Where the compounds described herein contain an ethylenic double bond or other geometrically chiral centers, unless otherwise specified, these compounds are intended to include E and Z geometrically isomers. All tautomers are also intended to be encompassed in this disclosure. Assay results may reflect data collected in racemic form, enantiomerically pure form, or any other form for stereochemistry. A single stereoisomer of a compound in this disclosure may, for example, substantially contain no other isomers, or may, for example, be as a racemic mixture, or mixed with all or other selected stereoisomers.

[0127] In some examples, the formula (I) compound having one or more chiral centers is enantiomerized.

[0128] This disclosure includes all geometric and positional isomers. For example, if the compound of formula (I) contains a double bond or a fused ring, both the cis and trans forms, as well as mixtures thereof, are included within the scope of this disclosure. If the compound contains a double bond, the substituent may be in the E or Z configuration unless otherwise specified. If the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may be in the cis or trans configuration unless otherwise specified.

[0129] In some embodiments, the content of this disclosure further includes compounds of formula (I) given by formula (II), [ka] Or relating to pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, or tautomers thereof.

[0130] In some embodiments, the present disclosure relates to a compound of formula (II) selected from the group consisting of compound (1) and compound (2). [ka] , Or relating to pharmaceutically acceptable salts, hydrates, solvates, or tautomers thereof.

[0131] In some embodiments, the present disclosure relates to compound (1) [ka] , Or relating to pharmaceutically acceptable salts, hydrates, solvates, or tautomers thereof.

[0132] In some embodiments, the present disclosure relates to compound (2) [ka] , Or relating to pharmaceutically acceptable salts, hydrates, solvates, or tautomers thereof.

[0133] In some embodiments, the present disclosure relates to a mixture of compound (1) and compound (2).

[0134] In some embodiments, the present disclosure is provided for the following: (S)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide, and, (R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide, Or relating to compounds selected from pharmaceutically acceptable salts, hydrates, solvates, or tautomers thereof.

[0135] In some embodiments, the present disclosure relates to (S)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide, or pharmaceutically acceptable salts, hydrates, solvates, or tautomers thereof.

[0136] In some embodiments, the present disclosure relates to (R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide, or pharmaceutically acceptable salts, hydrates, solvates or tautomers thereof.

[0137] In some embodiments, the present disclosure relates to a mixture of (S)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazin-1-yl)benzamide and (R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazin-1-yl)benzamide, or a pharmaceutically acceptable salt, hydrate, solvate or tautomer thereof.

[0138] The present disclosure includes any compound of formula (I) that is isotopically labeled (i.e., radioactively labeled) by replacing one or more atoms with atoms having different atomic masses or mass numbers. Examples of isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, and chlorine, for example, ²H (or deuterium (D)) respectively, 3 ³H, 11 ¹¹C, 13 ¹²C, 14 ¹³C, 15 ¹⁴N, 18 ¹⁵O, 17 ¹⁶O, 35 ³⁵S and 36 ³⁶Cl, for example, 3 ³H, 11 ¹⁴C and 14 ¹¹C. In one embodiment, there is provided a composition wherein substantially all of the atoms at positions within the compound of formula (I) are replaced with atoms having different atomic masses or mass numbers. In another embodiment, there is provided a composition wherein some atoms at positions within the compound of formula (I) are replaced, i.e., the compound of formula (I) is enriched at positions of atoms having different atomic masses or mass numbers. Isotopically labeled compounds of formula (I) can be prepared by methods known in the art.

[0139] This disclosure encompasses the preparation and use of salts of the compound of formula (I), including non-toxic, pharmaceutically acceptable salts. Examples of pharmaceutically acceptable addition salts include inorganic and organic acid addition salts and basic salts. Pharmaceutically acceptable salts include, but are not limited to, metal salts such as sodium salts, potassium salts, and cesium salts; alkaline earth metal salts such as calcium salts and magnesium salts; organic amine salts such as triethylamine salts, pyridine salts, picoline salts, ethanolamine salts, triethanolamine salts, dicyclohexylamine salts, and N,N'-dibenzylethylenediamine; inorganic salts such as hydrochloride salts, hydrobromide salts, phosphate salts, and sulfate salts; organic salts such as citrate salts, lactate salts, tartrate salts, maleate salts, fumarate salts, mandelate salts, acetate salts, dichloroacetate salts, trifluoroacetate salts, oxalate salts, and formate salts; sulfonates such as methanesulfonate salts, benzenesulfonate salts, and p-toluenesulfonate salts; and amino acid salts such as arginine salts, aspartate salts, and glutamate salts.

[0140] If the compound of the present invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired base (soda or a suitable inert solvent). Salts derived from pharmaceutically acceptable inorganic bases include aluminum, ammonium, calcium, copper, iron(III), iron(II), lithium, magnesium, manganese, potassium, sodium, and zinc. Pharmacologically acceptable salts derived from organic bases include salts of primary, secondary, tertiary, and quaternary amines, including substituted amines, cyclic amines, and naturally occurring amines. Examples include arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, and tromethamine.

[0141] If the compound of the present invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired acid (pure acid or a suitable inert solvent). Acids suitable for the preparation of pharmaceutically acceptable acid addition salts include acetic acid, ascorbic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glucuronic acid, glutamic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, lactobionic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucoic acid, naphthalenesulfonic acid, nicotinic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, and p-toluenesulfonic acid.

[0142] The compounds of this disclosure can form acid addition salts or base addition salts that may be pharmaceutically acceptable salts.

[0143] This disclosure encompasses the preparation and use of solvates of compounds of formula (I). Solvates can function as pharmacological equivalents because they typically do not significantly alter the physiological activity or toxicity of the compound. Compounds of formula (I) may exist in solvated form with pharmaceutically acceptable solvents (e.g., water, methanol, ethanol, etc.), and this disclosure is intended to include both solvated and unsolvated forms of compounds of formula (I).

[0144] In some examples, the solvates are hydrates. "Hydrate" refers to a specific subgroup of solvates in which the solvent molecule is water. Solvates can usually function as pharmacological equivalents. The preparation of solvates is well known in this field. For example, see M. Caira et al., J. Pharmaceut. Sci. [Journal of Pharmaceutical Sciences], 93(3):601-611 (2004), which describes the preparation of the solvate of fluconazole with ethyl acetate and water. Similar preparations of solvates, semi-solvates, hydrates, etc., are described in EC van Tonder et al., AAPS Pharm. Sci. Tech. [AAPS Pharmaceutical Science and Technology], 5(1): Article 12 (2004) and ALBingham et al., Chem. Commun. [Chemical Communications] 603-604 (2001). A typical, non-limiting process for preparing solvates involves dissolving the compound of formula (I) in a desired solvent (organic, water, or a mixture thereof) at a temperature between 20°C and approximately 25°C, cooling the solution at a rate sufficient to form crystals, and isolating the crystals by known methods (e.g., filtration). The presence of the solvent in the solvate crystals can be confirmed using analytical techniques such as infrared spectroscopy.

[0145] Compounds of formula (I) can modulate the activity of Bcl-2 family proteins. In some examples, compounds of formula (I) are used to modulate Bcl-2 and / or The activity of Bcl-xL can be regulated. In some examples, the compound of formula (I) can inhibit Bcl-2 and / or Bcl-xL.

[0146] Compound (1), (S)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide can modulate the activity of Bcl-2 family proteins. In some examples, compound (1) can modulate the activity of Bcl-2 and / or Bcl-xL. In some examples, compound (1) can inhibit Bcl-2 and / or Bcl-xL.

[0147] Compound (2), (R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide can modulate the activity of Bcl-2 family proteins. In some examples, compound (2) can modulate the activity of Bcl-2 and / or Bcl-xL. In some examples, compound (2) can inhibit Bcl-2 and / or Bcl-xL.

[0148] This disclosure involves finding a compound of formula (I) and its pharmaceutically acceptable salts, solvates, hydrates, stereoisomers and tautomers, or drug compositions comprising a compound of formula (I) that can be used to treat diseases or disorders related to the activity of Bcl-2 family proteins, such as proliferative disorders like systemic lupus erythematosus.

[0149] This disclosure involves finding compound (1), (S)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide, and pharmaceutically acceptable salts, solvates, hydrates, isomers and tautomers thereof, or drug compositions comprising compound (1), which can be used to treat diseases or disorders associated with the activity of Bcl-2 family proteins, such as proliferative disorders like systemic lupus erythematosus.

[0150] The disclosure includes finding compound (2), (R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide, and pharmaceutically acceptable salts, solvates, hydrates, stereoisomers and tautomers thereof, or drug compositions comprising compound (2), which can be used to treat diseases or disorders associated with the activity of Bcl-2 family proteins, such as proliferative disorders like systemic lupus erythematosus.

[0151] Method for preparing compounds as disclosed herein The compounds of this disclosure can be prepared by various methods (including standard chemistry). Suitable synthetic routes are described in the examples given below.

[0152] The compounds of this disclosure, i.e., compounds of formula (I), or their pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, or tautomers, can be prepared by methods known in the field of organic synthesis, as partially described by the synthetic schemes described in the examples. It is well understood that in the schemes described below, protecting groups for sensitive or reactive groups may be used, if necessary, according to general principles or chemistry. Protecting groups are handled according to standard methods of organic synthesis (TW Greene and PGMWuts, “Protective Groups in Organic Synthesis”, 3rd edition, Wiley, New York 1999). These groups are removed at a convenient stage in the compound synthesis using methods obvious to those skilled in the art. The selection process, reaction conditions, and their sequence should be consistent with the preparation of compounds having formula (I).

[0153] Those skilled in the art will recognize the presence of a stereocenter in the compound of formula (I). Therefore, this disclosure includes two possible stereoisomers (unless otherwise indicated and / or specified in the synthesis), and includes not only racemic compounds but also enantiomers and / or non-enantiomers. Unless otherwise specified, where a compound is required as a single enantiomer or non-enantiomer, it can be obtained by stereospecific synthesis or by the resolution of the final product or any convenient intermediate. The resolution of the final product, intermediate, or starting materials may be influenced by any suitable method known in the art. See, for example, “Stereochemistry of Organic Compounds” by ELEliel, S.H.Wilen, and L.M.Wander (Wiley-Interscience, 1994).

[0154] In view of this disclosure, the compounds of this disclosure are prepared by methods known to those skilled in the art, or by explanatory methods set forth in the following general scheme. For example, a method for preparing the compounds of this disclosure is disclosed in U.S. Patent No. 10,221,174, which is incorporated herein by reference in whole.

[0155] In the general schemes 1 to 4 shown below, Y, R 2 and R 4a It is defined as follows:

number

[0156] General Scheme 1 [ka] In general scheme 1, the presence of a base (e.g., triethylamine) causes compound A to be converted to R 4a Compound B is obtained by reacting it with NH2.

[0157] General Scheme 2 [ka] In general scheme 2, methyl 4-bromo-2-fluorobenzoate is reacted with compound C to obtain compound D, and the ester of compound D is hydrolyzed to obtain compound E. Compound E is coupled with compound B from general scheme 1 to obtain compound F.

[0158] General Scheme 3 [ka] In general scheme 3, compound G is converted to compound H.

[0159] General Scheme 4 [ka] In general scheme 4, compound H from general scheme 3 is reacted with Boc-protected piperidine to obtain compound J, and the Boc group is removed to obtain compound K. Compound K is reacted with 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-fluorobenzoate methyl to obtain compound L, and the ester of compound L is hydrolyzed to obtain compound M. Compound M is coupled with compound B from general scheme 1 to obtain compound (I), where E is a nitrogen atom, and here, TIFF0007914089000074.tif35 has a single bond.

[0160] General Scheme 5 [ka] In general scheme 5, compound H from general scheme 3 is reacted with 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine to obtain compound I. Compound I is coupled with compound F from general scheme 2 to obtain compound (I), where E is a carbon atom, and here, TIFF0007914089000076.tif35 has a double bond.

[0161] Methods using the compounds disclosed herein Since the compounds of formula (I) are inhibitors of Bcl-2 proteins (e.g., Bcl-2 and / or Bcl-xL), in some examples these compounds are useful tool compounds for studying in vitro or in vivo processes mediated by Bcl-2 proteins. In vitro, tool compounds of formula (I) can be used to study the effects of Bcl-2 family protein inhibition on purified proteins, cell extracts, complete cells and cell line models, etc. In vivo, tool compounds of formula (I) can be used to study the effects of Bcl-2 family protein inhibition in xenografts derived from cell lines, xenografts derived from patients, knock-in mouse models, etc.

[0162] Since the compounds of formula (I) are inhibitors of the Bcl-2 protein (e.g., Bcl-2 and / or Bcl-xL), many diseases, conditions, or disorders mediated by the Bcl-2 protein can be treated or prevented by administering these compounds to subjects. Therefore, this disclosure relates, as a whole, to a method for treating or preventing a disease, condition, or disorder in response to inhibition of the Bcl-2 protein (e.g., Bcl-2 and / or Bcl-xL) in animals that are suffering from or at risk of suffering from such disease, condition, or disorder, comprising administering an effective amount of one or more of the compounds of this disclosure to the animals.

[0163] In one example, the compound disclosed herein contains less than 10 μM of Bcl-2 and / or Bcl-xL IC. 50 In another example, the compound of the present disclosure has a concentration of less than 5 μM of Bcl-2 and / or Bcl-xL IC. 50 In another embodiment, the compound of the present disclosure is less than 1 μM of Bcl-2 and / or Bcl-xL IC. 50 In another example, the compound of the present disclosure is less than 0.5 μM of Bcl-2 and / or Bcl-xL IC. 50 In another example, the compound of the present disclosure has a concentration of less than approximately 0.1 μM of Bcl-2 and / or Bcl-xL IC. 50 In another example, the compound of the present disclosure has a concentration of less than approximately 0.05 μM of Bcl-2 and / or Bcl-xL IC. 50 In another example, the compound of the present disclosure has a concentration of less than approximately 0.025 μM of Bcl-2 and / or Bcl-xL IC. 50 It has. In another example, the compound of the present disclosure has a concentration of less than approximately 0.010 μM of Bcl-2 and / or Bcl-xL IC. 50 In another example, the compound of the present disclosure has a concentration of less than approximately 0.005 μM of Bcl-2 and / or Bcl-xL IC. 50 In another example, the compound of the present disclosure has a concentration of less than approximately 0.0025 μM of Bcl-2 and / or Bcl-xL IC. 50In another example, the compound of the present disclosure has a concentration of less than approximately 0.001 μM of Bcl-2 and / or Bcl-xL IC. 50 It has.

[0164] The disclosure further relates to a method for inhibiting Bcl-2 family proteins in animals that require it, for example, humans, the method comprising administering a therapeutically effective amount of at least one disclosed compound to the animal. In another embodiment, the disclosure relates to a method for inhibiting Bcl-2 family proteins in animals that require it, for example, humans, the method comprising administering a therapeutically effective amount of a drug composition comprising at least one compound of the disclosure to the animal.

[0165] The present disclosure further relates to a method for inhibiting Bcl-2 in an animal that requires it, for example, a human, the method comprising administering a therapeutically effective amount of at least one compound of the present disclosure to the animal. In another embodiment, the present disclosure relates to a method for inhibiting Bcl-2 in an animal that requires it, for example, a human, the method comprising administering a therapeutically effective amount of a drug composition comprising at least one compound of the present disclosure to the animal.

[0166] The Disclosure further relates to a method for inhibiting Bcl-xL in an animal that requires it, for example, a human, the method comprising administering a therapeutically effective amount of at least one compound of the Disclosure to the animal. In another embodiment, the Disclosure relates to a method for inhibiting Bcl-xL in an animal that requires it, for example, a human, the method comprising administering a therapeutically effective amount of a drug composition comprising at least one compound of the Disclosure to the animal.

[0167] In one embodiment, the Disclosure provides a method for treating or preventing a proliferative disorder in a subject, for example, a human, the method comprising administering a therapeutically effective dose of a compound of the Disclosure or a drug composition comprising at least one compound of the Disclosure. In several examples, the Disclosure provides a method for treating, preventing or improving an autoimmune disorder in a subject, for example, a human, the method comprising administering a therapeutically effective dose of a compound of the Disclosure or a drug composition comprising at least one compound of the Disclosure.

[0168] This disclosure provides a method for treating or improving a subject, for example, a human with systemic lupus erythematosus, the method comprising administering a therapeutically effective dose of the compounds of this disclosure or a drug composition comprising at least one of the compounds of this disclosure to a patient in need thereof. In one example, the patient is further diagnosed with lupus nephritis. In one example, the patient is diagnosed with elevated levels of circulating anti-dsDNA antibodies.

[0169] The compounds of this disclosure may be administered to a subject in an unpossible chemical form in the absence of any other components. The compounds of this disclosure may further be administered to a subject as part of a drug composition comprising the compounds in combination with one or more suitable pharmaceutically acceptable carriers. Such carriers may be selected from pharmaceutically acceptable excipients and adjuvants. The terms “pharmaceutically acceptable carrier” or “pharmaceutically acceptable vehicle” encompass any standard pharmaceutically acceptable carrier, solvent, surfactant, or vehicle. Suitable pharmaceutically acceptable vehicles include aqueous and non-aqueous vehicles. Standard pharmaceutically acceptable carriers and their formulations are described in Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 19th edition, 1995.

[0170] The drug compositions within the scope of this disclosure include all compositions in which the compounds of this disclosure are combined with one or more pharmaceutically acceptable carriers. In one example, the compounds of this disclosure are present in the composition in an amount effective to achieve the intended therapeutic purpose. Individual needs may differ, but the optimal range for determining the effective amount of each compound is within the technical scope of the art. Typically, the compounds of this disclosure can be administered orally daily to mammals, such as humans, in doses of about 0.0025 to about 1500 mg per kg of mammalian body weight, or equivalent amounts of pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, or tautomers, to treat specific disorders. The effective oral dose of the compounds of this disclosure administered to mammals is about 0.0025 to about 200 mg per kg of mammalian body weight, or equivalent amounts of pharmaceutically acceptable salts or solvates. For intramuscular injection, the dose is usually about half of the oral dose. In some examples, the compounds of the present disclosure or their pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, or tautomers are administered once daily.

[0171] The unit oral dose may contain about 0.01 mg to about 1 g of the compound disclosed herein, for example, about 0.01 mg to about 500 mg, about 0.01 mg to about 250 mg, about 0.01 mg to about 100 mg, 0.01 mg to about 50 mg, for example, about 0.1 mg to about 10 mg of the compound. The unit dose may be administered once or more times a day in the form of one or more tablets or capsules, each containing about 0.01 mg to about 1 g of the compound, or an equivalent amount thereof of a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, or tautomer.

[0172] In some embodiments, a drug composition comprising a compound of the Disclosure or at least one compound of the Disclosure is administered to a patient in need in a dose sufficient to produce one or more effects selected from the group consisting of: a decrease in the patient's urinary protein excretion; a decrease in the patient's serum anti-dsDNA autoantibody levels; a decrease in the patient's skin injury severity; a decrease in the patient's lymphadenopathy severity; a decrease in the patient's glomerulonephritis severity; a decrease in the patient's vasculitis severity; a decrease in the lymphocyte count in peripheral blood mononuclear cells (PMBCs) collected from the patient; a decrease in the lymphocyte count in the patient's spleen; and a decrease in the patient's renal lymphocyte infiltration.

[0173] In one embodiment, a drug composition comprising a compound of the present disclosure or at least one compound of the present disclosure is administered to a patient in need in a dose sufficient to reduce the patient's urinary protein excretion.

[0174] In one embodiment, a drug composition comprising a compound of the present disclosure or at least one compound of the present disclosure is administered to a patient in need in a dose sufficient to reduce the patient's serum anti-dsDNA autoantibody level.

[0175] In one embodiment, a drug composition comprising a compound of the present disclosure or at least one compound of the present disclosure is administered to a patient in need in a dose sufficient to reduce the severity of the patient's skin injury.

[0176] In one embodiment, a drug composition comprising a compound of the present disclosure or at least one compound of the present disclosure is administered to a patient in need in a dose sufficient to reduce the severity of the patient's lymphadenopathy.

[0177] In one embodiment, a drug composition comprising a compound of the present disclosure or at least one compound of the present disclosure is administered to a patient in need in a dose sufficient to reduce the severity of the patient's glomerulonephritis.

[0178] In one embodiment, a drug composition comprising a compound of the present disclosure or at least one compound of the present disclosure is used to provide a patient in need of the patient interstitial It is administered in a dose sufficient to reduce the severity of nephritis.

[0179] In one embodiment, a drug composition comprising a compound of the present disclosure or at least one compound of the present disclosure is administered to a patient in need in a dose sufficient to reduce the severity of the patient's vasculitis.

[0180] In one embodiment, a drug composition comprising a compound of the present disclosure or at least one compound of the present disclosure is administered to a patient in need in a dose sufficient to reduce the lymphocyte count in a group of peripheral blood mononuclear cells (PMBCs) collected from the patient.

[0181] In one embodiment, a drug composition comprising one of the compounds of the Disclosure or at least one of the compounds of the Disclosure is administered to a patient in need in a dose sufficient to reduce the number of lymphocytes in the patient's spleen.

[0182] In one embodiment, a drug composition comprising a compound of the present disclosure or at least one compound of the present disclosure is administered to a patient in need in a dose sufficient to reduce lymphocyte infiltration in the patient's kidneys.

[0183] The compounds of this disclosure or drug compositions comprising the compounds of this disclosure may be administered to any patient or subject who may benefit from the beneficial effects of the compounds of this disclosure. Such patients or subjects are, most importantly, mammals, such as humans and companion animals, but this disclosure is not intended to be limited thereto. In one example, the patient or subject is a human.

[0184] The compounds of this disclosure or drug compositions comprising the compounds of this disclosure may be administered in any form that achieves their desired purpose. For example, they may be administered orally, extra-gastrointestinal, subcutaneously, intravenously, intramuscularly, intraperitoneally, percutaneously, intranasally, transmucosally, rectally, intravaginally or orally, or by inhalation. The dosage and route of administration should be determined based on the specific subject's condition, taking into consideration factors such as the recipient's age, sex, health and weight, the condition or disorder being treated, the type of synchronized treatment (if any), the frequency of treatment, and the nature of the desired effect.

[0185] In one embodiment, the compound of the Disclosure or a drug composition containing the compound of the Disclosure may be administered orally. In another embodiment, the drug composition of the Disclosure may be administered orally and formulated into a tablet, sugar-coated tablet, capsule, or oral liquid formulation. In one embodiment, the oral formulation comprises extruded polyparticles containing the compound of the Disclosure.

[0186] Alternatively, the compounds of the Disclosure or drug compositions comprising the compounds of the Disclosure may be administered rectally and incorporated into suppositories.

[0187] Alternatively, the compounds of the Disclosure or drug compositions comprising the compounds of the Disclosure may be administered by injection.

[0188] Alternatively, the compounds of the Disclosure or drug compositions comprising the compounds of the Disclosure may be administered transdermally.

[0189] Alternatively, the compounds of the Disclosure or drug compositions comprising the compounds of the Disclosure may be administered by inhalation, intranasal administration, or transmucosal administration.

[0190] Alternatively, the compounds of the Disclosure or drug compositions comprising the compounds of the Disclosure may be administered via the vaginal route.

[0191] The drug compositions of the Disclosure may contain, on a weight basis, about 0.01% to 99%, for example, about 0.25% to 75% of the Compounds of the Disclosure, for example, about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, or about 75% of the Compounds of the Disclosure.

[0192] The drug compositions of this disclosure may comprise one or more compounds of formula (I). In some examples, the drug composition comprises compound 1 and compound 2 having the above structure, or a pharmaceutically acceptable salt, hydrate, solvate, or tautomer thereof. In one example, the drug composition comprises compound 1 having the above structure or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof, having a purity of at least 90%, wherein the composition comprises less than 10%, for example, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% of compound 2 having the above structure.

[0193] The drug compositions of this disclosure are prepared in forms that are publicly known based on this disclosure, for example, by general mixing, granulation, sugar-coated tablet manufacturing, dissolution, extrusion, or freeze-drying processes. Thus, drug compositions for oral use can be prepared by combining an active compound with a solid excipient, optionally grinding the resulting mixture to process the particulate mixture, and adding appropriate excipients (if there is a need or it is required) to obtain tablets or sugar-coated tablet cores.

[0194] Suitable excipients include sugars (e.g., lactose, sucrose, mannitol, or sorbitol), cellulose preparations, fillers such as calcium phosphate (e.g., tricalcium phosphate or calcium hydrogen phosphate), and thickeners such as starch paste (e.g., corn starch, wheat starch, rice starch, or potato starch), gelatin, tragacanth gum, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone. If necessary, one or more disintegrants such as the above starches and carboxymethyl starch, cross-linked polyvinylpyrrolidone, agar, or alginic acid or sodium alginate salts thereof may be added.

[0195] The adjuvants are typically fluidity modifiers and lubricants, such as silica, talc, stearic acid or its salts (e.g., magnesium stearate or calcium stearate) and polyethylene glycol. The sugar-coated tablet core has a suitable coating that is resistant to gastric acid. For this purpose, a concentrated sugar solution may be used, which may optionally contain gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and / or titanium dioxide, a lacquer solution and a suitable organic solvent or solvent mixture. To produce a gastric acid-resistant coating, a suitable cellulose formulation, such as a solution of acetylcellulose phthalate or hydroxypropyl methylcellulose phthalate, may be used. Dyes or pigments may be added to the tablet or sugar-coated tablet coating to identify or characterize combinations of doses of the active compound.

[0196] Other examples of orally administered drug formulations include push-fit capsules made of gelatin, or soft-seal capsules made of gelatin and a plasticizer such as glycerin or sorbitol. Push-fit capsules may contain particulate compounds or extruded multi-part compounds that can be mixed with a filler such as lactose, an adhesive such as starch and / or a lubricant such as talc or magnesium stearate and an optional stabilizer. In soft-seal capsules, the active compound is preferably dissolved or suspended in a suitable liquid (e.g., fatty oil or liquid paraffin). A stabilizer may also be added.

[0197] Possible drug formulations for rectal administration include, for example, suppositories consisting of a combination of one or more active compounds and a suppository matrix. Suitable suppository matrices include natural or synthetic triglycerides and paraffinic hydrocarbons. Gelatin rectal capsules consisting of a combination of an active compound and a matrix material such as liquid triglycerides, polyethylene glycol, or paraffinic hydrocarbons can also be used.

[0198] Formulations suitable for parenteral administration include aqueous solutions of the active compound in a water-soluble form, such as water-soluble salts, basic solutions, or acidic solutions. Alternatively, suspensions of the active compound may be prepared as oily suspensions. Suitable lipophilic solvents or vehicles used in suspensions may include fatty oils (e.g., sesame oil), synthetic fatty acid esters (e.g., ethyl oleate), triglycerides, or polyethylene glycol, such as polyethylene glycol-400 (PEG-400). The aqueous suspension may contain one or more substances, such as sodium carboxymethylcellulose, sorbitol, and / or dextran, to increase the viscosity of the suspension. The suspension may optionally contain stabilizers.

[0199] In another embodiment, the Disclosure provides a kit comprising a compound of the Disclosure (or a drug composition comprising a compound of the Disclosure), the kit being packaged in a form advantageous for use in practicing the method of the Disclosure. In one embodiment, the kit comprises a compound of the Disclosure (or a drug composition comprising a compound of the Disclosure) packaged in a container, e.g., a sealed bottle or container, and having a label attached to the container or included in the kit, the label describing the use of the compound or composition for practicing the method of the Disclosure. In one embodiment, the compound or composition is packaged in unit dosage forms. The kit may further include a device suitable for administering the composition according to a desired route of administration.

[0200] In other embodiments, the compound of the Disclosure or a drug composition containing the compound of the Disclosure is administered to a subject in combination with a second therapeutic agent. The second therapeutic agent is different from the compound of the Disclosure. The compound of the Disclosure and the second therapeutic agent may be administered simultaneously or sequentially to achieve the desired effect. In some embodiments, the second therapeutic agent is administered before the compound of the Disclosure or a drug composition containing the compound of the Disclosure. In some embodiments, the second therapeutic agent is administered after the compound of the Disclosure or a drug composition containing the compound of the Disclosure. In some embodiments, the second therapeutic agent is administered simultaneously with the compound of the Disclosure or a drug composition containing the compound of the Disclosure. The compound of the Disclosure and the second therapeutic agent may be administered as a single composition or as two separate compositions.

[0201] The second therapeutic agent is administered in an amount that provides the desired therapeutic effect. The effective dose range for each second therapeutic agent is known in the art, and the second therapeutic agent is administered to individuals in need within this established range.

[0202] The compound of the present disclosure and the second therapeutic agent may be administered together as a single unit dose, or separately as multiple unit doses, wherein the compound of the present disclosure may be administered before the second therapeutic agent, or vice versa. One or more doses of the compound of the present disclosure and / or one or more doses of the second therapeutic agent may be administered. Thus, the compound of the present disclosure may be used in combination with one or more second therapeutic agents, such as autoimmune disorder treatment agents, but are not limited to these.

[0203] In some examples, the second therapeutic agent is an autoimmune disorder treatment agent.

[0204] In some embodiments, the second therapeutic agent is a treatment for systemic lupus erythematosus. For example, the treatment for systemic lupus erythematosus may include, but is not limited to, nonsteroidal anti-inflammatory drugs, antimalarial drugs, steroids, immunosuppressants, and dehydroepiandrosterone (DHEA).

[0205] Non-steroidal anti-inflammatory drugs (NSAIDs) used to treat systemic lupus erythematosus include, but are not limited to, celecoxib, diflunisal, etodolac, ibuprofen, indomethacin, meloxicam, midrin, nabumetone, naproxen, oxaprozin, piroxicam, salicylsalicylic acid, sulindac, tolmetin, magnesium choline trisalicylate, and ketoprofen.

[0206] Non-exclusive examples of antimalarial drugs used to treat systemic lupus erythematosus include hydroxychloroquine, chloroquine, and quinacrine.

[0207] Non-exclusive examples of steroids used as treatments for systemic lupus erythematosus include prednisone, prednisolone, hydrocortisone, methylprednisolone, dexamethasone, triamcinolone, and topical steroids.

[0208] Non-exclusive examples of immunosuppressants used to treat systemic lupus erythematosus include azathioprine, mycophenolate mofetil, cyclosporine, methotrexate, leflunomide, cyclophosphamide, chlorambucil, and nitrogen mustard.

[0209] As described in the art, the second therapeutic agent described above may be prepared and administered, and one or more of these may be used in combination with the compounds of the present disclosure.

[0210] example Example 1 - Synthesis of Intermediates Intermediate 1: Synthesis of 1-cyclobutylidenepropan-2-one [ka] To a toluene (200 ml) solution containing cyclobutanone (5.0 g, 71.4 mmol), 1-(triphenylphosphoranylidene)-2-propanone (22.7 g, 71.4 mmol) was added, and the mixture was refluxed overnight. The solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / hexane 1 / 10 to 1 / 5) to obtain 1-cyclobutylidenepropan-2-one (5.0 g) as a yellow oily substance. 1 H NMR (400MHz, CDCl3) δ 5.95-5.93(m,1H),3.19-3.13(m,2H),2.91-2.84(m,2H),2.21(s,3H),2.21-2.11(m,2H).

[0211] Intermediate 2: Spiro[3.5]nonan-6,8-zeon synthesis [ka] Sodium methoxide (41.4 g, 30% in methanol) was added to a methanol solution (150 ml) containing 1-cyclobutylidenepropan-2-one (23.1 g, 0.21 mol) and methyl malonate (30.3 g, 0.23 mol). The mixture was heated under reflux with nitrogen gas for 4 hours and concentrated. The resulting residue was hydrolyzed in 2N potassium hydroxide (200 ml) at 70°C for 4 hours. The mixture was extracted with ethyl acetate (100 ml) and then titrated with 1N hydrochloric acid to pH 3-5. The resulting solution was heated at 70°C for 5 hours and extracted with ethyl acetate (100 ml x 3). The mixed organic layer was dried on magnesium sulfate and concentrated to obtain spiro[3.5]nonane-6,8-dione (19.8 g, 62.3%) as a yellow solid. This product was used directly in the next step without further purification. 1 H NMR (400MHz, DMSO-d6) δ 11.05 (s, 1H), 5.17 (s, 1H), 2.50-2.35 (m, 4H), 1.92-1.79 (m, 2H), 1.79-1.72 (m, 4H).

[0212] Intermediate 3: 8-Isobutoxyspiro[3.5]non-7-en-6-one [ka] To a toluene (150 ml) solution containing spiro[3.5]nonane-6,8-dione (19.8 g, 0.13 mol), 4-toluenesulfonic acid (248 mg, 0.0013 mol) and isobutanol (14.5 g, 0.2 mol) were added. The mixture was heated under reflux, and water was removed by azeotropic distillation. The solvent was removed under vacuum, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether 1 / 10-1 / 3) to obtain 8-isobutoxyspiro[3.5]non-7-en-6-one (25.0 g, 92.7%) as a yellow oil. 1 H NMR(400MHz, CDCl3)δ 5.31(s,1H),3.59(d,J=6.8Hz,2H),2.51(s,2H),2.45(s,2H),2.12-1.96(m,1H),1.93-1.83(m,6H),0.99(d,J=6.8Hz,6H).

[0213] Intermediate 4: Spiro[3.5]non-7-en-6-one synthesis [ka] At room temperature, Red-Al® (40 ml, 70% in toluene, 0.18 mol) was added dropwise to a toluene (100 mL) solution containing 8-isobutoxyspiro[3.5]non-7-en-6-one (25.0 g, 0.12 mol). The mixture was heated to 45°C for 4 hours and then quenched with 1N hydrochloric acid. The mixture was filtered, and the filtrate was concentrated and purified by silica gel column chromatography (ethyl acetate / petroleum ether 1 / 10) to obtain spiro[3.5]non-7-en-6-one (9.0 g, 55%) as a yellow oily substance.

[0214] Intermediate 5: Spiro[3.5]non-6-one synthesis [ka] Spiro[3.5]non-7-en-6-one (9.0 g) was hydrogenated with 1 atm hydrogen gas and catalyzed with a 10% Pd / C (1.0 g) solution in methanol (80 ml) for 5.5 hours. Pd / C was removed by filtration, and the filtrate was concentrated to obtain spiro[3.5]non-6-one (8.8 g, 96.4%) as a colorless oil, which was used directly in the next step without further purification. 1 H NMR (400MHz, CDCl3) δ 2.38 (s, 2H), 2.23 -2.20 (m, 2H), 1.89 -1.75 (m, 10H).

[0215] Intermediate 6: Synthesis of methyl 6-oxospiro[3.5]nonane-7-carboxylate [ka] At room temperature, methyl carbonate (28.7 g, 0.32 mol) was added to a suspension of sodium hydride (5.1 g, 0.13 mol) in tetrahydrofuran (150 mL), and then spiro[3.5]non-6-one in tetrahydrofuran (30 mL) was added. The mixture was refluxed for 2 hours. The reaction was quenched with saturated aqueous ammonium chloride solution and extracted with ethyl acetate (100 mL x 3). The mixed organic layer was washed with brine and concentrated. The resulting residue was purified by silica gel column chromatography to obtain methyl 6-oxospiro[3.5]nonane-7-carboxylate (4.0 g, 32%) as a pale yellow oil.

[0216] Intermediate 7: Synthesis of methyl 6-(((trifluoromethyl)sulfonyl)oxy)spiro[3.5]non-6-ene-7-carboxylate [ka] Potassium carbonate (5.6 g, 0.04 mol) and N,N-bis(trifluoromethylsulfonyl)aniline (7.9 g, 0.02 mol) were added to a solution of methyl 6-oxospiro[3.5]nonane-7-carboxylate (4.0 g, 0.02 mol) in tetrahydrofuran (25 mL). The mixture was stirred overnight at room temperature, diluted with water, and extracted with ethyl acetate (100 mL x 3). The mixed organic layer was washed with saturated brine, dried over magnesium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether 1 / 50-1 / 10) to obtain methyl 6-(((trifluoromethyl)sulfonyl)oxy)spiro[3.5]nonane-7-carboxylate (5.0 g, 76%) as a pale yellow oil.

[0217] Intermediate 8: Synthesis of methyl 6-(4-chlorophenyl)spiro[3.5]non-6-ene-7-carboxylate [ka] A mixture of methyl 6-(((trifluoromethyl)sulfonyl)oxy)spiro[3.5]non-6-ene-7-carboxylate (5.0 g, 0.015 mol), 4-chlorophenylboronic acid (2.58 g, 0.017 mol), CsF (4.63 g, 0.03 mol), and Pd(PPh3)4 (173 mg, 0.15 mol) in 1,2-dimethoxyethane (30 ml) and methanol (15 ml) was heated to 70°C for 2 hours with nitrogen gas. The solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether 1 / 10) to obtain methyl 6-(4-chlorophenyl)spiro[3.5]non-6-ene-7-carboxylate (4.0 g, 92%) as a colorless oil. 1 H NMR(400MHz,CDCl3)δ 7.30(d,J=8.5Hz,2H),7.06(d,J=8.5Hz,2H),3.48(s,3H),2.50-2.44(m,2H), 2.43(t,J=2.3(2.3 or 6.3?)Hz,2H),2.02-1.80(m,6H),1.74(t,J=6.3Hz,2H).

[0218] Intermediate 9: Synthesis of (6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methanol [ka] A solution of tetrahydrofuran containing methyl 6-(4-chlorophenyl)spiro[3.5]non-6-en-7-carboxylate (4.0 g, 0.014 mol) (20 mL) was mixed with a solution of tetrahydrofuran containing LiBH4 (910 mg, 0.042 mol) (10 mL). The mixture was stirred overnight at room temperature, quenched with 1N hydrochloric acid aqueous solution, and extracted with ethyl acetate (100 mL x 3). The mixed organic layer was washed with brine, dried over magnesium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether 1 / 10-1 / 3) to obtain (6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methanol (3.0 g, 81.7%) as a white solid. 1H NMR(400MHz,CDCl3)δ 7.31(d,J=8.4Hz,2H),7.09(d,J=8.4Hz,2H),3.93(d,J=4.2Hz,2H),2.37-2.26(m,2H),2.01-1.77(m,8H),1.74(t,J=6.3Hz,2H).

[0219] Intermediate 10: Synthesis of 7-(chloromethyl)-6-(4-chlorophenyl)spiro[3.5]non-6-ene [ka] Methylsulfonyl chloride (3.0 g, 0.026 mol) was added dropwise to a solution of (6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methanol (3.5 g, 0.013 mol) and trimethylamine (2.7 g, 0.026 mol) in dichloromethane (20 mL). The mixture was stirred at room temperature for 5 hours. The solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 7-(chloromethyl)-6-(4-chlorophenyl)spiro[3.5]non-6-en (2.75 g, 75.5%) as a yellow oil. 1 H NMR(400MHz, CDCl3)δ 7.31(d,J=8.4Hz,2H),7.09(d,J=8.5Hz,2H),3.93(s,2H),2.34-2.25(m,4H),1.97-1.78(m,6H),1.74(t,J=6.3Hz,2H).

[0220] Intermediate 11: Synthesis of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-ethyl bromobenzoate [ka] A mixture of 1-(triisopropylsilyl)-1H-pyrrolo[2,3-b]pyridine-5-ol (1.91 g), methyl 4-bromo-2-fluorobenzoate (1.70 g), and K3P04 (1.86 g) in diglyme (20 mL) was stirred at 115°C for 1 hour. The reaction was cooled, diluted with ethyl acetate (100 mL), washed with water, washed with brine, and concentrated. The residue was purified by silica gel chromatography (ethyl acetate / hexane 1 / 3) to obtain ethyl 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-bromobenzoate (1.8 g) as a white solid. 1 H NMR(400MHz,CDCl3)δ 9.28(s,1H),8.18(d,J=2.5Hz,1H),7.79(d,J=8.4Hz,1H),7.62(d,J=2.5Hz,1 H),7.40-6.96(m,2H),6.96(d,J=1.7Hz,1H),6.51-6.48(m,1H),3.89(s,3H).

[0221] Intermediate 12: Synthesis of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-bromobenzoic acid [ka] To a 10 mL solution of dioxane containing ethyl 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-bromobenzoate (300 mg, 0.867 mmol), 1 N NaOH (2.2 mL, 2.2 mmol) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was acidified with 1 N HCl, extracted with ethyl acetate, washed with brine, and dried over anhydrous MgSO4. By evaporation under reduced pressure, crude 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-bromobenzoic acid was obtained as a colorless oil. This product was used directly in the next step without further purification.

[0222] Intermediate 13: Synthesis of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-bromo-N-((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide [ka] To a 10 mL solution of DCM containing 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-bromobenzoic acid (100 mg, 0.3 mmol), 3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)benzenesulfonamide (95 mg, 0.3 mmol), DMAP (55 mg, 0.45 mmol), and EDCI (115 mg, 0.6 mmol) were added, and the mixture was stirred at room temperature for 24 hours. The solvent was removed under reduced pressure, and the residue was purified by silica gel chromatography (DCM / MeOH 95 / 5) to obtain 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-bromo-N-((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide (80 mg) as a yellow oil. MS m / z 630[M+H] + .

[0223] Intermediate 14: Synthesis of (S)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-bromobenzamide [ka] To a 10 mL solution of DCM containing 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-bromobenzoic acid (100 mg, 0.3 mmol), (S)-4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrobenzenesulfonamide (95 mg, 0.3 mmol), DMAP (55 mg, 0.45 mmol), and EDCI (115 mg, 0.6 mmol) were added, and the mixture was stirred at room temperature for 24 hours. The solvent was removed under reduced pressure, and the residue was purified by silica gel chromatography (DCM / MeOH 95 / 5) to obtain (S)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-bromobenzamide. 1 1H NMR (400MHz, DMSO-d6)δ 11.79(s,1H),8.59-8.52(m,2H),8.05(d,J=2.6Hz,1H),7.85(dd,J=9.2,2.4Hz ,1H),7.66(d,J=2.6Hz,1H),7.59-7.49(m,1H),7.48(d,J=8.2Hz,1H),7.34(dd, J=8.2,1.8Hz,1H),7.12(d,J=9.2Hz,1H),6.88(d,J=1.8Hz,1H),6.50-6.40(m,1 H),3.83-3.37(m,2H),3.72-3.56(m,2H),3.56-3.42(m,2H),3.37-3.01(m,3H).

[0224] Intermediate 15: Synthesis of (R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-bromobenzamide [ka] (R)-4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrobenzenesulfonamide (95 mg, 0.3 mmol), DMAP (55 mg, 0.45 mmol), and EDCI (115 mg, 0.6 mmol) were added to a 10 mL solution of DCM containing 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-bromobenzoic acid (100 mg, 0.3 mmol), and the mixture was stirred at room temperature for 24 hours. The solvent was removed under reduced pressure, and the residue was purified by silica gel chromatography (DCM / MeOH 95 / 5) to obtain (R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-bromobenzamide. 1 H NMR(400MHz,DMSO-d6)δ 11.79(s,1H),8.59-8.52(m,2H),8.05(d,J=2.6Hz,1H),7.85(dd,J=9.2,2.4Hz,1H),7.66 (d,J=2.6Hz,1H),7.59-7.49(m,1H),7.48(d,J=8.2Hz,1H),7.34(dd,J=8.2,1.8Hz,1H),7 .12(d,J=9.2Hz,1H),6.88(d,J=1.8Hz,1H),6.50-6.40(m,1H),3.83-3.37(m,2H),3.72-3.56(m,2H),3.56-3.42(m,2H),3.37-3.01(m,3H).

[0225] Intermediate 16: Synthesis of tert-butyl-4-(3-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(ethoxycarbonyl)phenyl)piperazine-1-carboxylate [ka] In dimethyl sulfoxide, a mixture of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-fluorobenzoate methyl (2.1 g, 7 mmol), N-Boc-piperazine (2.61 g, 0.014 mol), and dipotassium hydrogen phosphate (2.44 g, 0.014 mol) was heated overnight at 135°C. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL x 3). The mixed organic layer was washed with brine, concentrated and purified by silica gel column chromatography to obtain 4-(3-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(ethoxycarbonyl)phenyl)piperazine-1-carboxylate (2.4 g, 73%) as a white solid. 1 H NMR(400MHz,CDCl3)δ 9.42(br s,1H),8.20(d,J=2.5Hz,1H),7.95(d,J=8.9Hz,1H),7.53(d,J=2.5Hz,1H),7.37(dd,J=3.5,2.5Hz,1H),6.66(dd,J=8.9,2.5Hz,1H),6.46(d d,J=3.5,2.0Hz,1H),6.36(d,J=2.5Hz,1H),4.28(q,J=7.1Hz,2H),3.55-3.50(m,4H),3.21-3.17(m,4H),1.47(s,9H),1.26(t,J=7.1Hz,3H).

[0226] Intermediate 17: Synthesis of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(piperazine-1-yl)methyl benzoate [ka] 4-(3-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(ethoxycarbonyl)phenyl)piperazine-1-carboxylate (2.1 g) was added to a 10 mL solution of dichloromethane, and trifluoroacetic acid (6 mL) was added. The mixture was stirred at room temperature for 3 hours. The solvent was removed under reduced pressure, and crude 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(piperazine-1-yl)methyl benzoate (2.5 g) was used directly in the next step without further purification.

[0227] Intermediate 18: Synthesis of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)methyl benzoate [ka] Potassium carbonate (1.26 g, 9 mmol), potassium iodide (100 mg, 0.6 mmol), and 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(piperazin-1-yl)methyl benzoate (1.53 g, 3 mmol) were added to a solution of N,N-dimethylformamide (10 mL) containing 7-(chloromethyl)-6-(4-chlorophenyl)spiro[3.5]non-6-ene (851 mg, 3.3 mmol). The mixture was stirred overnight at room temperature. The reaction mixture was diluted with water and extracted with ethyl acetate (50 mL x 3). The mixed organic layer was washed with brine, concentrated and purified by silica gel column chromatography (ethyl acetate / petroleum ether 1 / 5-1 / 1) to obtain 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)methyl benzoate (1.3 g, 71%) as a white solid. 1 H NMR(400MHz,CDCl3)δ 9.98(s,1H),8.20(d,J=2.6Hz,1H),7.91(d,J=9.0Hz,1H),7.51(d,J=2.6Hz,1H),7.38(t,J= 3.5Hz,1H),7.28(d,J=8.3Hz,2H),6.97(d,J=8.3Hz,2H),6.62(dd,J=9.0,2.5Hz,1H),6.45( dd,J=3.5,2.0Hz,1H),6.32(d,J=2.5Hz,1H),4.26(q,J=7.1Hz,2H),3.20-3.12(m,4H),2.77 (s,2H),2.31-2.17(m,8H),1.98-1.72(m,6H),1.68(t,J=6.3Hz,2H),1.25(t,J=7.1Hz,3H).

[0228] Intermediate 19: Synthesis of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzoic acid [ka] A solution of 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)methyl benzoate (1.3 g, 2.1 mmol) and dioxane (15 mL) containing 2 N potassium hydroxide (12 mL, 0.042 mol) was heated overnight at 60°C. The mixture was neutralized to pH 7 with 1 N hydrochloric acid aqueous solution and extracted with ethyl acetate (50 mL x 3). The mixed organic layers were washed with brine, dried over magnesium sulfate, and concentrated to obtain 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzoic acid (1.1 g, 88.7%) as a white solid. 1 H NMR(400MHz,CDCl3)δ 10.34(s,1H),8.19(d,J=2.6Hz,1H),8.02(d,J=9.0Hz,1H),7.63(d,J=2.6Hz,1H), 7.38-7.34(m,1H),7.27(d,J=8.3Hz,2H),6.96(d,J=8.3Hz,2H),6.63(dd,J=9.0,2. 4Hz,1H),6.44(dd,J=3.5,1.5Hz,1H),6.22(d,J=2.4Hz,1H),3.81(s,2H),3.17-3.1 0(m,4H),2.80(s,2H),2.30-2.20(m,6H),1.98-1.72(m,6H),1.67(t,J=6.3Hz,2H).

[0229] Intermediate 20: Synthesis of 1-(oxetan-3-ylidene)propan-2-one [ka] 1-(triphenylphosphoranylidene)propan-2-one (98.6 g, 0.31 mol) was added to a 300 ml solution of DCM containing oxetane-3-one (20.6 g, 0.28 mol). The mixture was stirred overnight at room temperature. The DCM was removed under reduced pressure until the solid precipitated. The solid was removed by filtration, and the filtrate was concentrated and purified by silica gel column chromatography (ethyl acetate / heptane 1 / 5-1 / 3) to obtain 1-(oxetane-3-ylidene)propan-2-one (23.3 g, 74.3%) as a yellow oil.

[0230] Intermediate 21: Synthesis of 2-oxaspiro[3.5]nonane-6,8-zion [ka] Sodium methoxide (41.3 g, 30% MeOH solution) was added to a methanol (150 ml) solution containing 1-(oxetane-3-ylidene)propan-2-one (23.3 g, 0.21 mol) and methyl malonate (30.2 g, 0.23 mol). The mixture was heated under reflux with nitrogen gas for 1 hour. The solvent was removed under reduced pressure to obtain 6-hydroxy-8-oxo-2-oxaspiro[3.5]non-6-ene-5-carboxylate, which was used directly in the next step without further purification. 6-hydroxy-8-oxo-2-oxaspiro[3.5]non-6-ene-5-carboxylate was added to an aqueous solution of KOH (2 mol / L, 200 ml). The mixture was stirred at room temperature for 30 minutes, and the aqueous solution was extracted with ethyl acetate (150 ml x 3). The aqueous layer was adjusted to pH 3-5 with 1N hydrochloric acid and heated at 50°C for 4 hours. Water was removed under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 2-oxaspiro[3.5]nonane-6,8-dione (2.5 g, 77.0%) as a pale yellow oil. This product was used directly in the next step without further purification.

[0231] Intermediate 22: Synthesis of 8-isobutoxy-2-oxaspiro[3.5]non-7-en-6-one [ka] TsOH (238 mg, 0.0016 mol) and isobutanol (18 g, 0.24 mol) were added to a toluene (150 ml) solution containing 2-oxaspiro[3.5]nonane-6,8-dione (25 g, 0.16 mol). After stirring at room temperature for 1 hour, the reaction was completed. The solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether 1 / 5-1 / 3) to obtain 8-isobutoxy-2-oxaspiro[3.5]non-7-en-6-one (6 g, 43%) as a pale yellow oil. 1 H NMR(400MHz,CDCl3)δ 5.34(s,1H),4.47(d,J=6.1Hz,2H),4.45(d,J=6.1Hz,2H),3.60(d,J=6.8Hz 2H),2.80(s,2H),2.68(s,2H),2.09-2.01(m,1H),0.98(d,J=6.8Hz,6H).

[0232] Intermediate 23: Synthesis of 2-oxaspiro[3.5]non-7-en-6-one [ka] Red-Al® (40.4 g, 70% in toluene) was added dropwise to a toluene (100 ml) solution containing 8-isobutoxy-2-oxaspiro[3.5]non-7-en-6-one (14.7 g, 0.07 mol). The mixture was monitored at 45°C for 2 hours and quenched with 1 N HCl solution. The mixture was concentrated and purified by silica gel column chromatography (ethyl acetate / petroleum ether 1 / 10-1 / 5) to obtain 2-oxaspiro[3.5]non-7-en-6-one (8.8 g, 91%) as a colorless oil. This product was used directly in the next step without further purification.

[0233] Intermediate 24: Synthesis of 2-oxaspiro[3.5]non-6-one [ka] Pd / C (1 g) was added to a solution of tetrahydrofuran (80 ml) containing 2-oxaspiro[3.5]non-7-en-6-one (8.8 g). The mixture was hydrogenated with 1 atm of hydrogen gas for 2 hours at room temperature. After the reaction was complete, Pd / C was removed by filtration, and the solution was concentrated to obtain 2-oxaspiro[3.5]non-6-one (8.0 g, 89.6%) as a colorless oil. This product was used directly in the next step without further purification.

[0234] Intermediate 25: Synthesis of 6-oxo-2-oxaspiro[3,5]nonane-7-carboxylate [ka] The methyl carbonate (25.7 g, 0.28 mol) sodium hydride (4.6 g, 0.11 mol) was added dropwise to a tetrahydrofuran (150 ml) suspension using nitrogen gas. After the addition was complete, the mixture was heated under reflux. Then, a solution of 2-oxaspiro[3.5]non-6-one (11.2 g, 0.057 mol) in tetrahydrofuran (30 ml) was added. The reaction was heated under reflux for 2 hours, quenched with saturated ammonium chloride aqueous solution, and extracted with ethyl acetate (100 ml x 3). The mixed organic layer was washed with brine, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 6-oxo-2-oxaspiro[3.5]nonane-7-carboxylate (4.5 g, 69%) as a colorless oil. This product was used directly in the next step without further purification.

[0235] Intermediate 26: Synthesis of 6-(((trifluoromethyl)sulfonyl)oxy)-2-oxaspiro[3.5]non-6-ene-7-carboxylate [ka] N,N-bis(trifluoromethylsulfonyl)aniline (8.9 g, 0.02 mol) was added to a suspension of 6-oxo-2-oxaspiro[3.5]nonane-7-carboxylate (4.5 g, 0.02 mol) and potassium carbonate (6.3 g, 0.046 mol) in DMF (30 ml). The mixture was stirred overnight at room temperature, diluted with water, and extracted with ethyl acetate (100 ml x 3). The mixed organic layer was washed with brine, dried over MgSO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether 1 / 10-1 / 3) to obtain 6-(((trifluoromethyl)sulfonyl)oxy)-2-oxaspiro[3.5]non-6-ene-7-carboxylate (6.6 g, 86%) as a pale yellow oil. This product was used directly in the next step without further purification.

[0236] Intermediate 27: Synthesis of 6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-ene-7-carboxylate [ka] 4-chlorophenylboronic acid (3.13 g, 0.02 mol), CsF (6.08 g, 0.04 mol), and Pd(PPh3)4 (231 mg, 0.2 mmol) were added to a solution of 1,2-dimethoxyethane (30 ml) containing 6-(((trifluoromethyl)sulfonyl)oxy)-2-oxaspiro[3.5]non-6-ene-7-carboxylate (6.6 g, 0.02 mol) and methanol (10 ml). The mixture was heated to 70°C for 30 min under nitrogen gas. The solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether 1 / 5-1 / 3) to obtain 6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-ene-7-carboxylate (5.1 g, 87.3%) as a pale yellow solid. 1H NMR(400MHz,CDCl3)δ 7.33(d,J=8.4Hz,2H),7.07(d,J=8.4Hz,2H),4.54(d,J=5.6Hz,2H),4.48(d,J=5.6 Hz,2H),3.48(s,3H),2.74-2.70(m,2H),2.55-2.50(m,2H),2.04(t,J=6.4Hz,2H).

[0237] Intermediate 28: Synthesis of (6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methanol [ka] At room temperature, LiBH4 (475 mg, 0.022 mol) in tetrahydrofuran (10 ml) was added dropwise to a solution of tetrahydrofuran (20 ml) containing 6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-carboxylate (2.1 g, 0.0072 mol). The mixture was stirred at room temperature for 4 hours, quenched with 1 N HCl solution, and extracted with ethyl acetate (100 ml x 3). The mixed organic layer was washed with brine, dried over MgSO4, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether 1 / 5-1 / 1) to obtain (6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methanol (1.5 g, 78.9%) as a white solid. 1 H NMR(400MHz,CDCl3)δ 7.34(d,J=8.4Hz,2H),7.07(d,2H,J=8.4Hz),4.54(d,2H,J=6.0Hz),4.46(d,2H ,J=5.6Hz),3.93(s,2H),2.62(s,2H),2.40-2.33(m,2H),2.03(t,2H,J=6.4Hz).

[0238] Intermediate 29: Synthesis of 7-(chloromethyl)-6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-ene [ka] (6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methanol (1.5 g, 5.7 mmol) and triethylamine (836 mg, 8.6 mmol)-containing dichloromethane (15 ml) were mixed with methylsulfonyl chloride (980 mg, 8.6 mmol), and the mixture was stirred at room temperature for 3.5 hours. The solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 7-(chloromethyl)-6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en (1.4 g, 87.0%) as a white solid. 1 H NMR(400MHz,CDCl3)δ 7.35(d,2H,J=8.4Hz),7.16(d,2H,J=8.4Hz),4.53(d,2H,J=6.0Hz),4.45(d,2H ,J=5.6Hz),3.86(s,2H),2.64(s,2H),2.40-2.33(m,2H),2.03(t,2H,J=6.4Hz).

[0239] Intermediate 30: Synthesis of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)methyl benzoate [ka] To a 10 ml solution of DMF containing 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(piperazin-1-yl)methyl benzoate (382 mg, 0.82 mmol), 7-(chloromethyl)-6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-ene (200 mg, 0.75 mmol), potassium carbonate (310 mg, 2.25 mmol), DIPEA (290 mg, 2.25 mmol), and potassium iodide (24.9 mg, 0.15 mmol) were added, and the mixture was stirred overnight at room temperature. The mixture was diluted with water and extracted with ethyl acetate (50 ml x 3). The mixed organic layer was washed with brine, concentrated, and purified by silica gel column chromatography (ethyl acetate / petroleum ether 1 / 5-1 / 1) to obtain 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methyl)piperazin-1-yl)methyl benzoate (370 mg, 80.6%) as a white solid. MS m / z 613[M+H] + .

[0240] Intermediate 31: Synthesis of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzoic acid [ka] To a 10 ml solution of dioxane containing 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)methyl benzoate (370 mg, 0.6 mmol), 2N potassium hydroxide (6 ml, 12 mmol) was added, and the mixture was stirred overnight at 60°C. The solution was neutralized to pH 7 with 1N hydrochloric acid and extracted with ethyl acetate (100 ml x 3). The mixed organic layer was washed with brine, dried over magnesium sulfate, and concentrated to obtain 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzoic acid (1.1 g, 88.7%) as a white solid. MS m / z 585[M+H] + .

[0241] Intermediate 32: Synthesis of 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(1-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperidine-4-yl)methyl benzoate [ka] To a 10 ml solution of N,N-dimethylformamide containing 7-(chloromethyl)-6-(4-chlorophenyl)spiro[3.5]non-6-ene (850 mg, 3.04 mmol), potassium carbonate (1.26 g, 2.2 mmol), potassium iodide (100 mg, 0.61 mmol), and 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(piperidine-4-yl)methyl benzoate (1.0 g, 3.34 mmol) were added, and the mixture was stirred overnight at room temperature. The mixture was then diluted with water and extracted with ethyl acetate. The mixed organic layer was washed with brine and concentrated. The obtained residue was purified by silica gel column chromatography to obtain 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(1-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperidine-4-yl)methyl benzoate (1.0 g, 55.2%) as a pale yellow solid. ¹H NMR (400 MHz, CDCl3) δ 9.39(br s,1H),8.19(d,J=2.6Hz,1H),7.87(d,J=8.1Hz,1H),7.57(d,J=2.6Hz,1H),7.3 9(dd,J=3.5,2.5Hz,1H),7.30-7.23(m,2H),7.04-6.93(m,3H),6.72(d,J=1.6H z,1H),6.49(dd,J=3.5,2.0Hz,1H),3.87(s,3H),2.81-2.75(m,2H),2.73-2.71 (m,2H),2.28(s,2H),2.25-2.15(m,2H),1.98-1.76(m,6H),1.75-1.51(m,9H).

[0242] Example 2: Synthesis of (R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide (compound 2) [ka]

[0243] 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzoic acid, (R)-4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrobenzenesulfonamide, EDCI was reacted overnight at room temperature with a mixture of 4-(N,N-dimethylamino)pyridine and dichloromethane, and water was added. The aqueous layer was extracted with dichloromethane. The mixed organic layers were washed with brine, concentrated and purified by silica gel chromatography to obtain (R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide. 1 H NMR(400MHz, methanol-d4)δ 8.66(d,J=2.4Hz,1H),7.99(d,J=2.4Hz,1H),7.84(dd,J=9.2,2.4Hz,1H),7.64(d,J=8.9Hz,1H),7.51(d,J=2.4Hz,2H),7. 45(d,J=3.3Hz,1H),7.37(d,J=8.4Hz,2H),7.10(d,J=8.4Hz,2H),6.94(d,J=9.2Hz,1H),6.76(dd,J=8.9,2.3Hz,1H),6.40 (d,J=3.3Hz,1H),6.36(d,J=2.3Hz,1H),3.87(dd,J=11.8,4.2Hz,3H),3.83-3.70(m,3H),3.67(s,2H),3.62(dd,J=11.7,2 .9Hz,1H),3.51-3.41(m,2H),3.40-3.35(m,1H),3.29(dq,J=3.2,1.6Hz,1H),2.41(s,2H),2.26(s,2H),2.00-1.77(m,6H).

[0244] Example 3: Synthesis of (S)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide (compound 1) [ka]

[0245] 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzoic acid, (S)-4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrobenzenesulfonamide, EDCI was reacted overnight at room temperature with a mixture of 4-(N,N-dimethylamino)pyridine and dichloromethane, and water was added. The aqueous layer was extracted with dichloromethane. The mixed organic layers were washed with brine, concentrated and purified by silica gel chromatography to obtain (S)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide. 1H NMR(400MHz, methanol-d4)δ 8.66(d,J=2.4Hz,1H),7.99(d,J=2.4Hz,1H),7.84(dd,J=9.2,2.4Hz,1H),7.64(d,J=8.9Hz,1H),7.51(d,J=2.4Hz,2H),7. 45(d,J=3.3Hz,1H),7.37(d,J=8.4Hz,2H),7.10(d,J=8.4Hz,2H),6.94(d,J=9.2Hz,1H),6.76(dd,J=8.9,2.3Hz,1H),6.40 (d,J=3.3Hz,1H),6.36(d,J=2.3Hz,1H),3.87(dd,J=11.8,4.2Hz,3H),3.83-3.70(m,3H),3.67(s,2H),3.62(dd,J=11.7,2 .9Hz,1H),3.51-3.41(m,2H),3.40-3.35(m,1H),3.29(dq,J=3.2,1.6Hz,1H),2.41(s,2H),2.26(s,2H),2.00-1.77(m,6H).

[0246] Example: Synthesis of 4-(R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(1-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)-1,2,3,6-tetrahydropyridine-4-yl)benzamide [ka]

[0247] (R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-bromobenzamide-containing 1,2-dimethoxyethane (10 ml) and water (1 ml) were mixed with 1-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine, Pd(dppf)Cl2 and K2CO3, and the mixture was stirred at 80°C for 12 hours. The reaction was cooled to room temperature and diluted with water. The mixture was extracted with ethyl acetate (30 ml x 3), dried over anhydrous MgSO4 and concentrated. The residue was C 18 The product was purified by reverse-phase preparative HPLC to obtain (R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(1-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)-1,2,3,6-tetrahydropyridine-4-yl)benzamide. 1 H NMR(400MHz, methanol-d4)δ 8.68(d,J=2.3Hz,1H),7.97(d,J=2.6Hz,1H),7.88(dd,J=9.3,2.3Hz,1H),7.63(d,J=8.2Hz,1H),7.50( d,J=2.6Hz,1H),7.46(d,J=3.5Hz,1H),7.30(d,J=8.4Hz,2H),7.16(dd,J=8.2,1.7Hz,1H),7.10(d,J=8. 4Hz,2H),6.94(d,J=9.3Hz,1H),6.85(d,J=1.7Hz,1H),6.41(d,J=3.5Hz,1H),5.94-5.90(m,1H),3.95- 3.40(m,14H),3.15-3.03(m,1H),2.68-2.45(m,2H),2.43(s,2H),2.30-2.20(m,2H),2.03-1.77(m,8H).

[0248] Example 5-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(1-((6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methyl)-1,2,3,6-tetrahydropyridine-4-yl)-N-((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide [ka]

[0249] The compound in question was prepared using a procedure similar to the program described in Example 4. 1 H NMR(400MHz, methanol-d4)δ 8.70(d,J=2.3Hz,1H),7.99(d,J=2.5Hz,1H),7.90(dd,J=9.2,2.3Hz,1H),7.61(d,J=8.2Hz,1H),7.57(d,J=2.5Hz,1H),7.48(d, J=3.5Hz,1H),7.31(d,J=8.4Hz,2H),7.20-7.10(m,3H),6.96(d,J=9.2Hz,1H),6.82(d,J=1.6Hz,1H),6.44(d,J=3.5Hz,1H),5.93 -5.86(m,1H),4.53(d,J=5.9Hz,2H),4.49(d,J=5.9Hz,2H),4.00-3.90(m,2H),3.77-3.33(m,7H),3.26(d,J=7.0Hz,2H),3.15-3. 00(m,1H),2.70-2.65(m,2H),2.63-2.25(m,4H),2.07(t,J=6.3Hz,2H),2.00-1.85(m,1H),1.75-1.65(m,2H),1.46-1.30(m,2H).

[0250] Example 6-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(1-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)-1,2,3,6-tetrahydropyridine-4-yl)-N-((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide [ka]

[0251] The compound in question was prepared using a procedure similar to the program described in Example 4. 1 H NMR(400MHz, methanol-d4)δ 8.71(t,J=1.9Hz,1H),8.00-7.95(m,1H),7.90(dd,J=9.3,1.9Hz,1H),7.63(dd,J=8.1,1.4Hz,1H),7.56-7.50(m ,1H),7.46(dd,J=3.5,1.4Hz,1H),7.33-7.26(m,2H),7.18-7.06(m,3H),6.96(dd,J=9.3,1.4Hz,1H),6.81(s,1H ),6.43(dd,J=3.5,1.5Hz,1H),5.93-5.86(m,1H),4.00-3.94(m,2H),3.83-3.36(m,7H),3.26(d,J=7.0Hz,2H),3 .10-3.04(m,1H),2.67-2.40(m,4H),2.30-2.24(m,2H),2.02-1.77(m,9H),1.74-1.67(m,2H),1.45-1.30(m,2H).

[0252] Example 7: Synthesis of (R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(1-((6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methyl)-1,2,3,6-tetrahydropyridine-4-yl)benzamide [ka]

[0253] The compound in question was prepared using a procedure similar to the program described in Example 4. 1H NMR(400MHz, methanol-d4)δ 8.68(d,J=2.3Hz,1H),7.99(d,J=2.5Hz,1H),7.89(dd,J=9.2,2.3Hz,1H),7.65(d,J=8.2Hz,1H),7.54( d,J=2.5Hz,1H),7.48(d,J=3.4Hz,1H),7.33(d,J=8.4Hz,2H),7.21-7.16(m,1H),7.13(d,J=8.4Hz,2H), 6.95(d,J=9.3Hz,1H),6.86(d,J=1.6Hz,1H),6.43(d,J=3.5Hz,1H),5.94-5.90(m,1H),4.60-4.43(m,4 H),3.95-3.40(m,14H),3.15-3.00(m,1H),2.80-2.60(m,4H),2.38-2.25(m,2H),2.08(t,J=6.3Hz,2H).

[0254] Example 8: Synthesis of (R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide [ka]

[0255] A mixture of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)-2-oxaspiro[3,5]non-6-en-7-yl)methyl)piperazine-1-yl)benzoic acid (290 mg, 0.5 mmol), (R)-4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrobenzenesulfonamide (236 mg, 0.75 mmol), EDCI (191 mg, 1 mmol), and 4-(N,N-dimethylamino)pyridine (591 mg, 0.75 mmol) in dichloromethane (15 ml) was stirred overnight at room temperature. The solvent was removed under vacuum, and the resulting residue was purified by silica gel column chromatography to obtain (R)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide (150 mg, 34.1%) as a yellow solid. 1 H NMR(400MHz, methanol-d4)δ 8.67(d,J=2.3Hz,1H),7.99(d,J=2.3Hz,1H),7.85(dd,J=9.3,2.3Hz,1H),7.64(d,J=8.8Hz,1H),7.5 2(d,J=2.3Hz,1H),7.45(d,J=3.5Hz,1H),7.39(d,J=8.4Hz,2H),7.13(d,J=8.4Hz,2H),6.95(d,J=9.3 Hz,1H),6.76(dd,J=8.8,2.4Hz,1H),6.41(d,J=3.5Hz,1H),6.34(d,J=2.4Hz,1H),4.54(d,J=5.9Hz,2 H),4.48(d,J=5.9Hz,2H),3.93-3.35(m,19H),2.70-2.65(m,2H),2.33(s,2H),2.08(t,J=6.3Hz,2H).

[0256] Example 9-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)-N-((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide [ka]

[0257] A mixture of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzoic acid (250 mg, 0.43 mmol), 3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)benzenesulfonamide (202 mg, 0.64 mmol), EDCI (164 mg, 0.86 mmol), and 4-(N,N-dimethylamino)pyridine (78 mg, 0.64 mmol) in dichloromethane (10 ml) was stirred overnight at room temperature and then concentrated. The resulting residue was purified by silica gel chromatography to obtain 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)-2-oxaspiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)-N-((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide (150 mg, 39.6%) as a yellow solid. 1H NMR(400MHz, methanol-d4)δ 8.70(d,J=2.3Hz,1H),8.01(d,J=2.6Hz,1H),7.87(dd,J=9.2,2.3Hz,1H),7.66(d,J=8.8Hz,1H),7.56(d,J=2.6Hz,1H),7.4 7(d,J=3.5Hz,1H),7.39(d,J=8.4Hz,2H),7.13(d,J=8.4Hz,2H),6.97(d,J=9.2Hz,1H),6.76(dd,J=8.8,2.4Hz,1H),6.43(d, J=3.5Hz,1H),6.32(d,J=2.4Hz,1H),4.54(d,J=5.9Hz,2H),4.48(d,J=5.9Hz,2H),4.03-3.94(m,2H),3.67(s,2H),3.55-3. 27(m,12H),2.69(s,2H),2.35-2.25(m,2H),2.08(t,J=6.3Hz,2H),2.05-1.93(m,1H),1.76-1.69(m,2H),1.45-1.35(m,2H).

[0258] Example 10: Synthesis of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)-N-((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide [ka]

[0259] A mixture of 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzoic acid (1.75 g, 3 mmol), 3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)benzenesulfonamide (1.43 g, 4.5 mmol), EDCI (1.15 g, 6 mmol), 4-(N,N-dimethylamino)pyridine (550 mg, 4.5 mmol), and dichloromethane (40 ml) was reacted overnight at room temperature, and then water was added. The aqueous layer was extracted with dichloromethane. The mixed organic layer was washed with brine, concentrated and purified by silica gel column chromatography to obtain 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazin-1-yl)-N-((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide (1.7 g, 64.4%) as a yellow solid. 1H NMR (400 MHz, methanol-d4)δ 8.70(d,J=2.3Hz,1H),8.01(d,J=2.7Hz,1H),7.87(d,J=9.2,2.3Hz,1H),7.66(d,J=8.9Hz,1H),7.5 5(d,J=2.7Hz,1H),7.47(d,J=3.4Hz,1H),7.38(d,J=8.4Hz,2H),7.10(d,J=8.4Hz,2H),6.97(d,J=9. 2Hz,1H),6.77(dd,J=8.9,2.4Hz,1H),6.44(d,J=3.4Hz,1H),6.34(d,J=2.4Hz,1H),4.02-3.94(m,3H) ),3.66(s,3H),3.49-3.38(m,2H),3.41-3.25(m,7H),2.42(s,3H),2.26(s,3H),2.00-1.67(m,4H),1 .45-1.38(m,2H).

[0260] Example 11: Synthesis of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)-N-((3-nitrophenyl)sulfonyl)benzamide [ka]

[0261] The compound in question was prepared using a procedure similar to the program described in Example 10. 1 H NMR(400MHz,DMSO-d6)δ 11.70(s,1H),9.47(s,1H),8.62(d,J=2.2Hz,1H),8.44(d,J=8.3Hz,1H),8.27(d,J=7.9Hz,1H) ),8.02-7.97(m,1H),7.84-7.75(m,1H),7.56-7.43(m,3H),7.40(d,J=8.3Hz,2H),7.11(d,J=8 .3Hz,2H),6.72(d,J=8.9Hz,1H),6.40-6.35(m,1H),6.30(s,1H),3.80-3.65(m,2H),3.55(s, 2H), 3.28-2.95 (m, 4H), 2.82-2.65 (m, 2H), 2.31 (s, 2H), 2.22-2.15 (m, 2H), 1.93-1.60 (m, 8H).

[0262] Example 12: Synthesis of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)-N-((4-(methylamino)-3-nitrophenyl)sulfonyl)benzamide [ka]

[0263] The compound in question was prepared using a procedure similar to the program described in Example 10. 1H NMR(400MHz, methanol-d4)δ 8.78(d,J=2.3Hz,1H),8.05(d,J=2.6Hz,1H),7.96(dd,J=9.2,2.3Hz,1H),7.80(d,J=8.9Hz,1H ),7.61(d,J=2.6Hz,1H),7.46(d,J=3.5Hz,1H),7.34(d,J=8.4Hz,2H),7.01(d,J=8.4Hz,2H),6. 90(d,J=9.2Hz,1H),6.68(dd,J=8.9,2.4Hz,1H),6.46(d,J=3.5Hz,1H),6.18(d,J=2.4Hz,1H),3 .60(s,2H),3.50-3.12(m,8H),3.06(s,3H),2.38(s,2H),2.30-2.16(m,2H),1.97-1.73(m,8H).

[0264] Example 13: Synthesis of 2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)-N-((4-(dimethylamino)-3-nitrophenyl)sulfonyl)benzamide [ka]

[0265] The compound in question was prepared using a procedure similar to the program described in Example 10. 1H NMR(400MHz, methanol-d4)δ 8.41(d,J=2.2Hz,1H),8.08(d,J=2.5Hz,1H),7.91(dd,J=9.4,2.3Hz,1H),7.81(d,J=8.9Hz, 1H),7.68(d,J=2.3Hz,1H),7.48(d,J=3.5Hz,1H),7.34(d,J=8.0Hz,2H),7.04(d,J=9.4Hz,1H ),7.01(d,J=8.0Hz,2H),6.71-6.63(m,1H),6.51(d,J=3.5Hz,1H),6.15(d,J=1.9Hz,1H),3.5 9(s,2H),3.52-3.20(m,8H),2.98(s,6H),2.38(s,2H),2.25-2.17(m,2H),1.96-1.72(m,8H).

[0266] Example 14: Synthesis of 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(1-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperidine-4-yl)-N-((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide [ka]

[0267] 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(1-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperidine-4-yl)benzoic acid (200 mg, 0.34 mmol), 3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)benzenesulfonamide (162 mg, 0.52 mmol), EDCI (130 mg, 0.68 mmol), 4-(N,N-dimethylamino)pyridine (63.4 mg, 0.52 mmol) l) The mixture containing dichloromethane (15 ml) was stirred overnight at room temperature and purified by silica gel column chromatography to obtain 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(1-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperidine-4-yl)-N-((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide (170 mg, 57.3%) as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ 11.94(s,1H),11.64(s,1H),8.50-8.42(m,2H),7.97(d,J=2.6Hz,1H),7 .76(dd,J=9.2,2.2Hz,1H),7.52-7.36(m,5H),7.11(d,J=7.9Hz,2H),6. 99(d,J=9.2Hz,1H),6.91-6.86(m,1H),6.55(s,1H),6.37(s,1H),3.89- 3.79 (m, 2H), 3.35-2.90 (m, 10H), 2.32-2.10 (m, 5H), 1.95-1.15 (m, 17H).

[0268] Example 15 - Bcl-2 and Bcl-X by compound (I) L Inhibition against The fluorescein-labeled BIM (81-106), BAK (72-87), and BID (79-99) peptides, named Flu-BIM, Flu-BAK, and Flu-BID respectively, were used with Bcl-2 and Bcl-X LIt was used as a fluorescent probe in the FP assay of Mcl-1. By monitoring the total fluorescence polarization value of a mixture consisting of a fixed-concentration fluorescent probe and protein whose concentration was increased to complete saturation, Flu-BIM and Bcl-X were used for Bcl-2. L Flu-BAK for and Flu-BID for Mcl-1 d The values ​​were determined to be 0.55±0.15 nM, 4.4±0.8 nM, and 6.9±0.9 nM, respectively. Fluorescence polarization values ​​were measured using an Infinite M-1000 plate reader (Tecan US, Research Triangle Park, NC) on a Microfluor 96-well, black round-bottom plate (Thermo Scientific). In each well, 1 nM of Flu-BIM, or 2 nM of Flu-BAK, or 2 nM of Flu-BID with increased concentrations of Bcl-2, or Bcl-X were measured. L Alternatively, Mcl-1 was added to 125 μL of final volume of assay buffer (100 mM potassium phosphate, pH 7.5, 100 μg / mL bovine γ-globulin, 0.02% sodium azide, Invitrogen, containing 0.01% Triton X-100 and 4% DMSO). The plate was mixed and incubated at room temperature for 1 hour, then gently shaken to ensure equilibrium. Polarization values ​​in millipolarization units (mP) were measured at excitation wavelength 485 nm and emission wavelength 530 nm. The equilibrium dissociation constant (K) was then determined by fitting the S-shaped dose-dependent FP increase as a function of protein concentration using Graphpad Prism 5.0 software (Graphpad Software, San Diego, California). d ) was calculated.

[0269] Bcl-2, Bcl-X LThe Ki values ​​of representative compounds of this disclosure against Bcl-1 were determined by competitive binding experiments using 96-well plates containing fixed concentrations of fluorescent probes and proteins in each well, with serial dilutions of the inhibitors added. A mixture of 5 μL of the tested inhibitor in DMSO and 120 μL of pre-incubated protein / probe complexes in assay buffer was added to the assay plate and incubated at room temperature for 2 hours with gentle shaking. The final concentrations of protein and probe were 1.5 nM and 1 nM for the Bcl-2 assay, 10 nM and 2 nM for the Bcl-xL assay, and 20 nM and 2 nM for the Mcl-1 assay, respectively. Negative controls (corresponding to 0% inhibition) containing only the protein / probe complex and positive controls (corresponding to 100% inhibition) containing only the free probe were included in each assay plate. Fluorescence polarization values ​​were measured as described above. IC 50 The values ​​were determined by nonlinear regression fitting of the competition curve. Measured IC 50 Based on the values, the probe values ​​relative to the protein, and the concentrations of the protein and probe in the competitive assay, the formula described in Nikolovska-Coleska et al., Analytical Biochemistry 332:261-73 (2004) is used to determine the K of the competitive inhibitor. i The value was calculated. Furthermore, using the formula from Huang, Journal of Biomolecular Screening 8:34-38 (2003), K d The value was calculated.

[0270] Table 2 shows the inhibitory activity of representative compounds of this disclosure against Bcl-2, Bcl-xL, and Mcl-1.

[0271] [Table 2]

[0272] Example 16 - MRL / LPR mouse model for SLE Female MRL / MpJ-Fas lpr / J mice (18-20g, 7-8 weeks old) were obtained from Jackson Laboratory. These mice were found to have spontaneous lymphoid proliferation mutations (Fas lpr These mice were homozygous for the gene and exhibited systemic autoimmunity, massive lymphadenopathy associated with abnormal T cell proliferation, arthritis, and immune complex glomerulonephropathy. Therefore, these mice are useful as a model in systemic lupus erythematosus (SLE) research.

[0273] Animals were housed and treated in a temperature-controlled environment with a 12-hour light / 12-hour dark cycle. A total of 36 mice were randomly divided into three groups based on urinary protein concentration and body weight, and treated for 10 weeks with either vehicle (po, qd), 30 mg / kg compound 1 (po, qd), or 100 mg / kg compound 1 (po, qd). The experimental scheme and program for the humane treatment and use of animals were approved by the Animal Experiment Committee (IACUC) of WuXiAppTec(Shanghai) Co., Ltd. (Shanghai, China).

[0274] Example 17 - Dose-dependent reduction of urinary protein levels in an SLE mouse model by compound 1 Using the Coomassie Brilliant Blue (CBB) assay and a urinary protein test kit (Nanjing Jiancheng Biotechnology Institute, Suzhou, China), urinary protein was evaluated individually in each mouse every two weeks. As shown in Figure 1, urinary protein levels in mice from the vehicle group increased from 304.4 ± 110.0 mg / L at week 8 to 1376.1 ± 391.4 mg / L at week 18. Treatment with compound 1 reduced urinary protein levels in a dose-dependent manner. Compound 1 at 30 mg / kg significantly inhibited the increase in urinary protein levels (p<0.05, compared to the vehicle group), and at completion of the study (week 18), 536.1 ± 146.8 mg / L of protein was detected in the urine. Furthermore, throughout the entire experimental process, compound 1 at 100 mg / kg maintained urinary protein levels (266.8 ± 37.7 mg / L at week 18; p < 0.0001, compared to the vehicle group). In short, compound 1 dose-dependently improved lupus nephritis in a mouse spontaneous SLE model.

[0275] Example 18 - Dose-dependent reduction of serum anti-dsDNA autoantibody levels in an SLE mouse model by compound 1. Anti-dsDNA autoantibody levels were measured by ELISA. In short, 96-well plates were coated overnight at 4°C with 10 μg / mL calf thymus ds-DNA (Sigma-Aldrich, Massachusetts, USA). They were washed with PBS containing 0.05% Tween® 20 (PBS-T), blocked with 1% bovine serum albumin, and then serum samples (4000x dilution) were added and incubated at room temperature for 1 hour. Goat anti-mouse IgG-HRP conjugate (InJay, California, USA) was added at room temperature for 1 hour, followed by three washes with PBS-T. Finally, Super TMB-ELISA substrate solution (Pierce Biotechnology, Illinois, USA) was added, incubated for 30 minutes, and the OD value at 405 nm was recorded after the reaction stopped. As shown in Figure 2, at the end of the study, serum anti-dsDNA autoantibody levels in MRL / lpr mice decreased to 76.8% (p<0.05, relative to the vehicle group) and 56.4% (p<0.0001, relative to the vehicle group) after treatment with 30 mg / kg compound 1 or 100 mg / kg compound 1, respectively. These results are consistent with the therapeutic effect of compound 1 in reducing urinary protein.

[0276] Example 19 - Dose-dependent reduction of skin injury severity in an SLE mouse model by compound 1 Starting from week 12, skin damage will be scored weekly using a scoring system, with a scale of 0 to 4 (0 - normal, 1 - small damage with a diameter of 2-4 mm, 2 - damaged area < 0.5 cm²). 2 ,3-damage area 0.5cm 2 ~1cm 2 , 4-damage area>1cm 2 As shown in Figure 3, MRL / lpr mice developed skin damage at week 13, and their pathological score reached 2.1 ± 0.2 at the end of the study (week 18). Treatment with compound 1 at 30 mg / kg and 100 mg / kg significantly reduced the skin damage score at week 18 (p<0.0001, compared to the vehicle group).

[0277] Example 20 - Dose-dependent reduction of lymphadenopathy in an SLE mouse model by compound 1. Throughout the study, lymph nodes were scored weekly using a scoring system based on lymph node diameter, ranging from 0 to 6 (0 - normal, 1 - <1 cm in one site, 2 - <1 cm in two sites, 3 - <1 cm in three sites, 4 - >1 cm in one site and <1 cm in two sites, 5 - >1 cm in two sites and <1 cm in one site, 6 - >1 cm in three sites). As shown in Figure 4, MRL / lpr mice developed lymph node swelling at week 9, and the pathological score reached 4.5 ± 0.4 at the end of the study (week 18). When mice were treated with compound 1 at 30 mg / kg and 100 mg / kg, respectively, the lymph node swelling score decreased to 2.7 ± 0.4 and 1.6 ± 0.4 (p < 0.0001, relative to the vehicle group).

[0278] Example 21 - Dose-dependent improvement in pathological score of lupus nephritis in a mouse model of SLE by compound 1. At the end of the study, the spleen and lymph nodes were collected and weighed. The kidneys were fixed in formalin, embedded in paraffin, sliced ​​into 5 μm thick slices, and stained with hematoxylin and eosin. The pathological score of the tissue was evaluated under a microscope using a blind method; please refer to the following.

[0279] The pathological scores of the tissues were evaluated under a microscope using a blind method. The membranous glomerulonephritis score (GN score) was evaluated on a scale of 0 to 4 (0 - normal, 1 - mild, localized or early proliferation, 2 - moderate or clear proliferation and matrix increase, 3 - diffuse and localized or diffuse proliferation, 4 - severe diffuse proliferation with crescents / sclerosis). For inflammation and necrosis, the renal interstitial nephritis score (IN score) was graded on a scale of 0 to 4 (0 - normal, 1 - incidental, localized or microsac mononuclear cells (MNCs, 10-14 cells), 2 - localized infiltration of MNCs (15-30 cells), 3 - multifocal and widespread infiltration of MNCs, 4 - severe infiltration of MNCs with widespread necrosis). The vasculitis score ranges from 0 to 4 points (0 - normal, 1 - incidental MNC perivascular infiltration, 2 - MNC perivascular infiltration without necrosis in some lesions, 3 - MNC multifocal perivascular infiltration with / without necrosis, 4 - MNC multifocal or diffuse perivascular infiltration with extensive necrosis).

[0280] As shown in Figure 5, MRL / lpr mice in the vehicle group developed moderate to severe glomerulonephritis, interstitial nephritis, and vasculitis at 18 weeks, which is indicated by a pathological score of approximately 3.0. Treatment with compound 1 dose-dependently reduced the pathological scores in all three aspects. Notably, statistical significance was achieved in both individual and overall scores when mice were treated with compound 1 at 100 mg / kg.

[0281] Example 22 - Significant reduction in lymphocyte counts in PMBC, spleen, and kidneys in an SLE mouse model using compound 1 At the end of the study (week 18), spleens and kidneys were obtained from mice and digested into single-cell suspensions. Red blood cells from whole blood were broken down to obtain peripheral blood mononuclear cells (PBMCs). The cells were stained with the following fluorescently labeled antibodies: PerCP-Cyanine5.5 conjugate anti-mouse CD8a (BD Biosciences), AF700 conjugate anti-mouse CD3 (BD ​​Biosciences), PE-eFluor610 conjugate anti-mouse B220 (BD Biosciences), e506 conjugate anti-mouse CD45 (BD Biosciences), BV605 conjugate anti-mouse CD138 (BioLegend), and BV711 conjugate anti-mouse CD4 (BioLegend).

[0282] Based on forward scattering (FCS) and side scattering (SSC), all cells were gated primarily to live lymphocytes. The samples were analyzed by flow cytometry (BD FACSCalibur, USA) to calculate the number of subtypes for each lymphocyte. As shown in Figures 6 and 7, after treatment with 100 mg / kg of compound 1, the number of total lymphocytes (CD45+), T cells (CD45+CD3+), CD4+ T cells (CD45+CD3+CD4+), CD8+ T cells (CD45+CD3+CD8+), double-negative T cells (CD45+CD3+CD4-CD8-), B cells (CD45+CD3-B220+), activated B cells (CD45+CD3-B220+CD69+), plasmablasts (CD45+CD3-B220+CD138+), plasma cells (CD45+CD3-B220-CD138+), and CD138+ cells (CD45+CD3-CD138+) were significantly reduced.

[0283] It should be noted that double-negative T cells were the most significantly reduced compared to other lymphocytes. While not theoretically bound, a decrease in autoantibody-secreting plasmablasts and plasma cells may explain the reduced serum anti-dsDNA autoantibody results and the observed reduction in lupus nephritis severity in Example 7. The number of lymphocytes infiltrating the kidney is shown in Figure 8. Treatment with 100 mg / kg of compound 1 significantly reduced the number of pro-inflammatory CD4+ T cells and double-negative T cells in the kidney. While not theoretically bound, a decrease in pro-inflammatory CD4+ and double-negative T cell infiltration may explain some of the effects observed in the treatment of lupus nephritis.

[0284] This disclosure enables those skilled in the art to manufacture and use the inventions herein based on several modified embodiments. Various variations, modifications, and improvements of this disclosure that those skilled in the art may conceive are part of this disclosure, including some variations, modifications, substitutions, and improvements. Therefore, the foregoing description illustrates the discoveries herein by example. The foregoing description and examples are examples of the invention and are not limiting. Therefore, the scope of the invention is described in the appended claims.

[0285] All patents and publications incorporated herein by reference are incorporated herein by reference in their entirety.

Claims

1. A pharmaceutical composition for treating systemic lupus erythematosus in patients, 【Chemistry 1】 A therapeutically effective amount of a compound selected from the group consisting of the above, or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof, The pharmaceutical composition.

2. The compound is 【Chemistry 2】 The pharmaceutical composition according to claim 1, or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof.

3. The compound is 【Transformation 3】 The pharmaceutical composition according to claim 1, or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof.

4. A pharmaceutical composition for treating systemic lupus erythematosus in a patient, comprising a therapeutically effective amount of a compound, wherein the compound is (S)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-(((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide.

5. The pharmaceutical composition according to any one of claims 1 to 4, wherein the patient is diagnosed with lupus nephritis.

6. The pharmaceutical composition according to any one of claims 1 to 5, administered to a patient in need thereof in a dose sufficient to produce one or more effects selected from the group consisting of: a decrease in the patient's urinary protein excretion; a decrease in the patient's serum anti-dsDNA autoantibody levels; a decrease in the patient's skin injury severity; a decrease in the patient's lymphadenopathy severity; a decrease in the patient's glomerulonephritis severity; a decrease in the patient's vasculitis severity; a decrease in the lymphocyte count in the peripheral blood mononuclear cell (PMBC) group taken from the patient; a decrease in the lymphocyte count in the patient's spleen; and a decrease in the patient's renal lymphocyte infiltration.

7. A pharmaceutical composition according to any one of claims 1 to 6, administered to a patient in need in a dose sufficient to reduce the excretion of protein in the patient's urine.

8. The pharmaceutical composition according to any one of claims 1 to 7, which is administered to a patient in need in a dose sufficient to reduce the patient's serum anti-dsDNA autoantibody level.

9. A pharmaceutical composition according to any one of claims 1 to 8, administered to a patient in need in a dose sufficient to reduce the severity of the patient's skin injury.

10. A pharmaceutical composition according to any one of claims 1 to 9, administered to a patient in need in a dose sufficient to reduce the severity of lymph node swelling in the patient.

11. The pharmaceutical composition according to any one of claims 1 to 10, which is administered to a patient in need in a dose sufficient to reduce the severity of the patient's glomerulonephritis.

12. The pharmaceutical composition according to any one of claims 1 to 11, which is administered to a patient in need in a dose sufficient to reduce the severity of the patient's interstitial nephritis.

13. The pharmaceutical composition according to any one of claims 1 to 12, which is administered to a patient in need in a dose sufficient to reduce the severity of the patient's vasculitis.

14. The pharmaceutical composition according to any one of claims 1 to 13, administered to a patient in need in a dose sufficient to reduce the number of lymphocytes in a group of peripheral blood mononuclear cells (PMBCs) collected from the patient.

15. The pharmaceutical composition according to any one of claims 1 to 14, which is administered to a patient in need in a dose sufficient to reduce the number of lymphocytes in the patient's spleen.

16. The pharmaceutical composition according to any one of claims 1 to 15, which is administered to a patient in need in a dose sufficient to reduce lymphocyte infiltration in the patient's kidney. 【Request Item 17】 【Chemistry 4】 The use of compounds selected from the group consisting of, or pharmaceutically acceptable salts, solvates, hydrates, or tautomers thereof, in the preparation of drugs for the treatment of systemic lupus erythematosus in patients requiring such drugs.

18. The compound is 【Transformation 5】 The use according to claim 17, or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof.

19. The compound is 【Transformation 6】 The use according to claim 17, or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof.

20. Use of (S)-N-((4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)sulfonyl)-2-((1H-pyrrolo[2,3-b]pyridine-5-yl)oxy)-4-(4-((6-(4-chlorophenyl)spiro[3.5]non-6-en-7-yl)methyl)piperazine-1-yl)benzamide in the preparation of a drug for the treatment of systemic lupus erythematosus in patients who require it.

21. The use according to any one of claims 17 to 20, wherein the patient is diagnosed with lupus nephritis.

22. The use according to any one of claims 17 to 21, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to produce one or more effects selected from the group consisting of: a decrease in the patient's urinary protein excretion; a decrease in the patient's serum anti-dsDNA autoantibody levels; a decrease in the patient's skin injury severity; a decrease in the patient's lymphadenopathy severity; a decrease in the patient's glomerulonephritis severity; a decrease in the patient's vasculitis severity; a decrease in the lymphocyte count in the peripheral blood mononuclear cell (PMBC) population taken from the patient; a decrease in the lymphocyte count in the patient's spleen; and a decrease in the patient's renal lymphocyte infiltration.

23. The use according to any one of claims 17 to 22, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the excretion of protein in the patient's urine.

24. The use according to any one of claims 17 to 23, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the patient's serum anti-dsDNA autoantibody level.

25. The use according to any one of claims 17 to 24, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the severity of the patient's skin injury.

26. The use according to any one of claims 17 to 25, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the severity of the patient's lymph node swelling.

27. The use according to any one of claims 17 to 26, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the severity of the patient's glomerulonephritis.

28. The use according to any one of claims 17 to 27, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the severity of the patient's interstitial nephritis.

29. The use according to any one of claims 17 to 28, wherein the compound or drug is administered to a patient in need in a dose sufficient to reduce the severity of the patient's vasculitis.

30. The use according to any one of claims 17 to 29, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the number of lymphocytes in a group of peripheral blood mononuclear cells (PMBCs) taken from the patient.

31. The use according to any one of claims 17 to 30, wherein the compound or drug composition is administered to a patient in need in a dose sufficient to reduce the number of lymphocytes in the patient's spleen.

32. The use according to any one of claims 17 to 31, wherein the compound drug composition is administered to a patient in need in a dose sufficient to reduce lymphocyte infiltration in the patient's kidneys.

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