Thiol isomerase inhibitors and methods of making and using same

Abstract

The present invention discloses novel thiol isomerase inhibitors and their use in the prevention or treatment of the onset or progression of diseases or conditions involving one or more extracellular thiol isomerase enzymes. Also disclosed is a method of preventing or treating thrombosis, thrombotic disease, platelet aggregation, fibrogenesis, infectious disease, viral disease, immunological disease, inflammation, neurological disease, neurodegenerative disease, cancer, or a combination thereof in a patient, the method comprising administering to the patient in need thereof a therapeutically effective amount of a compound.

Description

[0001] Cross - Reference to Related Applications

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 399,516, filed on August 19, 2022, and U.S. Provisional Application No. 63 / 454,974, filed on March 28, 2023. For all purposes, the entire contents of both applications are incorporated herein by reference.

[0003] Statement Regarding Federally Sponsored Research or Development

[0004] This invention was made with government support under R21 CA231000 and F31DE029661 awarded by the National Institutes of Health (NIH). The government has certain rights in the invention. Background Art

[0005] Thiol - isomerases are members of a large family of disulfide oxidoreductases that catalyze post - translational disulfide - bond exchange essential for the correct folding of newly synthesized proteins. There are approximately twenty members of the thiol - isomerase / disulfide - oxidoreductase large family in humans, and the domain composition of thiol - isomerases is a - b - b’ - a’. These thiol - isomerases are generally capable of redox and isomerization reactions and are often found in the endoplasmic reticulum, where they catalyze the correct folding of newly translated proteins.

[0006] In addition, some thiol - isomerases, such as protein disulfide isomerase (PDI), ERp5, ERp57, ERp72, and thioredoxin (TRX), have recently been found to have extracellular functions. These five thiol - isomerases (hereinafter referred to as extracellular thiol - isomerases) are secreted by cells such as platelets and re - attach to the plasma membrane, where they function as extracellular oxidoreductases. Extracellular thiol - isomerases have also been identified on the surface of endothelial cells and play roles in the activation of thrombosis and fibrin formation, as well as platelet aggregation, granule secretion, fibrinogen binding, and calcium mobilization. Among these enzymes, the role of PDI in thrombosis has been the most thoroughly studied and best understood, and ERp5, ERp57, and ERp72 are also considered essential.

[0007] Members of the PDI family are upregulated in many different types of cancer, including ovarian cancer, prostate cancer, lung cancer, melanoma, lymphoma, and glioma, and inhibition of PDI is cytotoxic to ovarian cancer cell lines. These enzymes are thought to be multifunctionally involved in the development of cancer, and they play roles in oncogene activation, avoidance of apoptosis, secretion of major histocompatibility complex class I - related A protein (MICA), and resistance to chemotherapeutic drugs.

[0008] Cancer patients have a significantly higher risk of developing arterial and venous thromboembolism, and this risk is substantially elevated in patients receiving systemic chemotherapy. The risk of death is doubled in cancer patients with thromboembolism, and thrombosis is the second leading cause of death in cancer patients. The annual mortality rates due to arterial or venous thrombosis in cancer patients are approximately 3-fold and 50-fold higher, respectively, than in the general population, and they can account for up to 14% of cancer mortality.

[0009] Thromboembolic events often complicate the treatment of cancer. Although venous thromboembolic events are more common, arterial thromboembolic events are also more prevalent in patients with malignancies than in other populations. However, there are significant differences in risk among individuals depending on cancer type, chemotherapy regimen, and other clinical risk factors such as cancer stage, catheter use, or other interventions (such as surgery). When deciding whether to prophylactically anticoagulate a patient, the risk of the patient having a thromboembolic event needs to be weighed against the risk of taking anticoagulant drugs. Considerable time and resources have been invested in developing risk models and scores (such as the Khorana score) to attempt to predict patients at highest risk of thromboembolic events. Unfortunately, recent studies have found that these risk models generally have poor predictive ability for the risk of venous thromboembolism (VTE). In addition, even if the risk score can be correctly predicted and applied to patients with venous thromboembolism, it is not suitable to use low molecular weight heparin or similar anticoagulant therapies to treat arterial thrombosis and its sequelae, such as myocardial infarction and stroke.

[0010] There is still a need in the art for novel compounds and methods for treating cancer, thrombosis, thrombotic diseases, and for treating or preventing cancer-induced thrombosis. Summary of the Invention

[0011] In one embodiment, a method of treating cancer or treating or preventing cancer-induced thrombosis comprises administering to a patient in need a therapeutically effective amount of a compound of formula (I), (II), (III) or a pharmaceutically acceptable salt thereof:

[0012]

[0013] Wherein,

[0014] Ar 1 is

[0015] R 1is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxyl, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 , where R 9 is hydrogen, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH or aryl; Q 1 and Q 2 are each independently a bond, O or NR 10 , and R 10 is hydrogen, C1-C8 alkyl, C1-C6 haloalkyl or C1-C8 alkyl-OH;

[0016] R 2 is hydrogen, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C1-C4 alkanoyl or unsubstituted or substituted aryl;

[0017] X is O, S or N;

[0018] Y is N or CH;

[0019] When X is N, R 6 is hydrogen, C1-C8 alkyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl or C1-C4 alkanoyl, and when X is S or O, R does not exist 6 ;

[0020] R 7 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxyl, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 , where Q 1 , Q 2 and R 9 are as defined above;

[0021] Ar 2 is unsubstituted or substituted by R 8A substituted aryl or an unsubstituted or R-substituted heteroaryl, where R 8 is NO2, cyano, halogen, NH2, COOH, hydroxyl, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 8 (C=O)Q 1 R 2 is as defined above; 9 where Q 1 and Q 2 and R 9 are as defined above;

[0022] R 4 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxyl, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 where Q 1 and Q 2 and R 9 are as defined above; and

[0023] R 5 is hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl or C1-C4 alkanoyl,

[0024] subject to the following conditions a) and b):

[0025] a) When Ar 1 is R 4 is -OMe, R 5 is hydrogen and Ar 2 is phenyl or phenyl substituted by R 8 for the compounds of formula (III), R 1 and R 2 and R 8 do not satisfy the following six conditions:

[0026]

[0027] b) When Ar 1 is R 1 is hydrogen, NO2, NH2, mono-C1-C8 alkylamino or di-C1-C8 alkylamino; R 2 is hydrogen or C1-C8 alkyl; R 4 is hydrogen or C1-C8 alkyl; and R 5 when it is hydrogen; then Ar 2 is not unsubstituted phenyl or phenyl substituted with C1-C8 alkyl at the 2-position.

[0028] In another embodiment, a compound of formula (I), (II), (III) or a pharmaceutically acceptable salt thereof:

[0029]

[0030] wherein,

[0031] Ar 1 is

[0032] R 1 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxy, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 , where R 9 is hydrogen, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH or aryl; Q 1 and Q 2 each independently is a bond, O or NR 10 , and R 10 is hydrogen, C1-C8 alkyl, C1-C6 haloalkyl or C1-C8 alkyl-OH;

[0033] R 2 is hydrogen, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C1-C4 alkanoyl or unsubstituted or substituted aryl;

[0034] X is O, S or N;

[0035] Y is N or CH;

[0036] when X is N, R 6is hydrogen, C1-C8 alkyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl or C1-C4 alkanoyl, and when X is S or O, R does not exist 6 ;

[0037] R 7 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxy, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 wherein Q 1 、Q 2 and R 9 are as defined above;

[0038] Ar 2 is unsubstituted or substituted aryl or unsubstituted or substituted heteroaryl;

[0039] R 4 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxy, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 wherein Q 1 、Q 2 and R 9 are as defined above; and

[0040] R 5 is hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl or C1-C4 alkanoyl,

[0041] subject to the following conditions a) and b):

[0042] a) When Ar 1 is R 4 is -OMe, R 5 is hydrogen and Ar 2 is phenyl or phenyl substituted by R 8 for the compounds of formula (III), R 1, R 2 and R 8 do not satisfy the following six conditions:

[0043]

[0044] b) When Ar 1 is R 1 is hydrogen, NO2, NH2, mono-C1-C8 alkylamino or di-C1-C8 alkylamino; R 2 is hydrogen or C1-C8 alkyl; R 4 is hydrogen or C1-C8 alkyl; and when R 5 is hydrogen, Ar 2 is not an unsubstituted phenyl or a phenyl substituted with a C1-C8 alkyl at the 2-position.

[0045] In yet another embodiment, a pharmaceutical composition comprises a compound of formula (I), (II), (III) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0046] The foregoing and other features are illustrated by the following drawings and detailed description.

[0047] Generally, the present disclosure may alternatively comprise any suitable components disclosed herein, consist of any suitable components disclosed herein, or consist essentially of any suitable components disclosed herein. The present disclosure may additionally or alternatively be formulated to be free or substantially free of any components, materials, ingredients, adjuvants or species used in prior art compositions, or free of any components, materials, ingredients, adjuvants or species that are not necessary for achieving the functions and / or purposes of the present disclosure.

[0048] BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Reference is now made to the drawings, which are exemplary embodiments and should not be considered limiting:

[0050] Figure 1A and Figure 1B : In the enzymatic thiol isomerase activity assay, the inhibitory effects of zileuton (ZAF, Figure 1A ) and montelukast (MON, Figure 1B ) on thiol isomerase activity.

[0051] Figure 2A and Figure 2B : Cytotoxicity assessment of analogs 21, 27, 28, ZAF and MON against the BEAS-2B cell line( Figure 2A ) and the HEK-293 cell line( Figure 2B , legend same as in Figure 2A . Controls included treatment with (TX, 1% v / v, positive control) and 1% DMSO (negative control) treatment. In cases where cell viability > 100% was observed, the data was presented as 100% cell viability (normalized data). The experiments were performed in quadruplicate.

[0052] Figure 3A and Figure 3B : Inhibitory effects of ZAF and compound 21 on mammalian cell growth. Cells were seeded and treated with the drugs after 24 hours. After an additional 24-hour incubation, cell viability was measured using Prestoblue. These graphs present the summary of results from at least 3 independent experiments for each cell line. The cell lines used included HEK-293 non-cancer cells (not shown); OVCAR-8 and HCT116. Compare the 3-day growth inhibitory effects of ZAF (circles) and compound 21 (inverted triangles) on OVCAR-8 ( Figure 3A ) and HCT116 cancer cells ( Figure 3B ).

[0053] Figure 4A and Figure 4B : Washed human platelets were incubated with compound 21 for five minutes before stimulation with collagen and then measured using an optical aggregometer.

[0054] Figure 5 : ZAF inhibits cancer cell-induced activation of the coagulation cascade. ZAF inhibits the production of factor Xa in HCT116 cells even at low concentrations.

[0055] Figure 6 : Insulin turbidity assay, the inhibitory effects of compound 32 on PDI, ERp5, ERp57, and ERp72 are similar to those of zafirlukast.

[0056] Figures 7A - 7D : Zafirlukast inhibits cell thiol isomerase activity. ( Figure 7A ) As measured by eosin-GSSG fluorescence, zafirlukast treatment inhibits cell thiol isomerase activity in a concentration-dependent manner (n = 4). Data are presented as mean ± standard deviation. One-way ANOVA and post hoc Dunnett's test, where *p = 0.0217 at 3 μM zafirlukast, **p = 0.0020 at 10 μM zafirlukast, and ****p < 0.0001 at 30 μM zafirlukast compared to the control. ( Figure 7B) In OVCAR8 cells, the levels of PDI, ERp5, ERp57, and ERp72 remained similar with increasing concentrations of zileuton (n = 3). Data are expressed as mean ± standard deviation. Student's t-test was used to compare with the control. No significant changes.( Figure 7C ) Montelukast (n = 3) and( Figure 7D ) Compound 32 analogs (n = 4) also inhibited cellular thiol isomerase activity. Data are expressed as mean ± standard deviation. One-way ANOVA and post hoc Dunnett's test, where *p = 0.0219 for 10 μM montelukast, **p = 0.0018 for 30 μM montelukast, ***p = 0.0002 for 100 μM montelukast, while *p = 0.0383 for 1 μM compound 32, **p = 0.0017 for 3 μM compound 32, ***p = 0.0004 for 10 μM compound 32, and ****p < 0.0001 for 30 μM compound 32, compared to the control.

[0057] Figure 8A and Figure 8B : Zileuton, montelukast, and compound 32 selectively induced cytotoxicity in cancer cells.( Figure 8A ) The cytotoxicity of zileuton was approximately 5-fold that of montelukast and approximately 1.5-fold that of compound 32 (n = 3).( Figure 8B ) Figure 6 、Figure 7 and Figure 8A Comparison of the relative potencies of each compound in the experiment. Detailed Description of the Invention

[0058] The present invention discloses novel thiol isomerase inhibitors and their therapeutic use in preventing or treating the development or progression of diseases or disorders involving one or more extracellular thiol isomerases, including protein disulfide isomerase (PDI), thioredoxin, ERp5, ERp57, ERp72, or combinations thereof. These compounds have potential use in preventing and / or treating cancer, thrombosis (arterial, venous), thrombotic diseases, or combinations thereof.

[0059] A promising mechanism against cancer-induced thrombosis is to utilize compounds that can inhibit thiol isomerases, which include protein disulfide isomerase (PDI) and closely related enzymes ERp5, ERp57, and ERp72. These enzymes have extracellular activities and play important roles in both arterial and venous thrombosis as well as in various cancers. PDI, ERp5, ERp57, and ERp72 are secreted by platelets; they attach to the plasma membrane and function as extracellular redox enzymes on the plasma membrane. They are essential for thrombosis, fibrin formation, and platelet aggregation, dense granule secretion, fibrinogen binding, and calcium mobilization. Among potential and existing antithrombotic drugs, the ability to simultaneously inhibit both arterial and venous thrombosis is unprecedented. Thiol isomerases are also upregulated in many different types of cancers, including ovarian cancer, prostate cancer, lung cancer, melanoma, lymphoma, and glioma, and an increase in thiol isomerase levels is positively correlated with carcinogenic transformation, gene transcription, and increased metastasis, and even promotes resistance to chemotherapy and radiotherapy. Similar to their role in platelets, they can also be secreted by tumor cells and function on the cell surface. Thiol isomerase inhibition is cytotoxic to multiple tumor types. For example, inhibiting thiol isomerases can significantly inhibit the growth of ovarian tumors in xenograft mice without causing toxicity to normal tissues.

[0060] Since thiol isomerase inhibitors have the potential dual use as anti-cancer and antithrombotic drugs, thiol isomerases are promising targets for preventing cancer-related thrombosis. Interestingly, despite their similarities, thiol isomerases are not redundant in arterial and venous thrombosis because inhibiting one thiol isomerase blocks thrombosis, and they have different roles in cancer growth and progression. Inhibitors of any one thiol isomerase or a pan-thiol isomerase inhibitor would be valuable. The US Food and Drug Administration (FDA)-approved drugs zileuton (ZAF) and related montelukast (MON) have been identified as promising broad-spectrum thiol isomerase inhibitors, and their potential in preventing and treating cancer-induced thrombosis has been demonstrated, validating the anti-tumor, anti-platelet, and anticoagulant functions of these two compounds.

[0061] The use of the compounds described herein can overcome two major weaknesses of current prophylactic anticoagulant therapies. Data indicate that, unlike any clinically used antithrombotic drug, ZAF can inhibit both arterial and venous thrombosis. Additionally, while both arterial and venous thrombosis are affected, hemostatic function is not affected, and thus the bleeding risk is not increased. Further, thiol isomerase inhibition has a cytotoxic effect on specific cancer cell lines as well as on an ovarian cancer xenograft model. Collectively, these data suggest that thiol isomerase inhibitors can significantly improve the treatment outcome of cancer-related thrombosis by targeting two major types of thrombosis without affecting bleeding time, thereby reducing the risks of current prophylactic therapies, while they can serve as anti-tumor drugs and form part of a chemotherapy regimen.

[0062] The present disclosure provides compounds of zafirlukast derivatives of formula (I), (II), (III) or pharmaceutically acceptable salts thereof:

[0063]

[0064] wherein

[0065] Ar 1 is

[0066] R 1 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxy, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 wherein R 9 is hydrogen, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH or aryl; Q 1 and Q 2 each independently is a bond, O or NR 10 and R 10 is hydrogen, C1-C8 alkyl, C1-C6 haloalkyl or C1-C8 alkyl-OH; specifically, R 1 is an electron-withdrawing group; more specifically, R 1 is NO2, cyano, C1-C6 haloalkyl, CF3 or C2-C6 alkanoyl;

[0067] R 2is hydrogen, a C1-C8 alkyl group, a C1-C6 haloalkyl group, a C1-C8 alkyl-OH group, a C3-C7 cycloalkyl group, a (C3-C7 cycloalkyl)C0-C6 alkyl group, a C1-C4 alkanoyl group or an unsubstituted or substituted aryl group; specifically, R 2 is hydrogen, a C1-C6 alkyl group, a C1-C8 alkyl-OH group or an unsubstituted or substituted phenyl group;

[0068] X is O, S or N;

[0069] Y is N or CH;

[0070] When X is N, R 6 is hydrogen, a C1-C8 alkyl group, a C3-C7 cycloalkyl group, a (C3-C7 cycloalkyl)C0-C6 alkyl group or a C1-C4 alkanoyl group, and when X is S or O, R does not exist 6 ;

[0071] R 7 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxyl, a C1-C8 alkyl group, a C1-C6 haloalkyl group, a C1-C8 alkyl-OH group, a C1-C8 alkyl-NH2 group, a C1-C8 alkoxy group, a mono-C1-C8 alkylamino group, a di-C1-C8 alkylamino group, a C2-C6 alkenyl group, a C3-C7 cycloalkyl group, a (C3-C7 cycloalkyl)C0-C6 alkyl group, a C2-C6 alkanoyl group or -Q 1 (C=O)Q 2 R 9 where Q 1 、Q 2 and R 9 are as defined above;

[0072] Ar 2 is an unsubstituted or substituted aryl group or an unsubstituted or substituted heteroaryl group; specifically, Ar 2 is an unsubstituted or R 8 substituted phenyl or naphthalene, where R 8 is NO2, cyano, halogen, NH2, COOH, hydroxyl, a C1-C8 alkyl group, a C1-C6 haloalkyl group, a C1-C8 alkyl-OH group, a C1-C8 alkyl-NH2 group, a C1-C8 alkoxy group, a mono-C1-C8 alkylamino group, a di-C1-C8 alkylamino group, a C2-C6 alkenyl group, a C3-C7 cycloalkyl group, a (C3-C7 cycloalkyl)C0-C6 alkyl group, a C2-C6 alkanoyl group or -Q 1 (C=O)Q 2 R 9 where Q 1 、Q 2 and R 9 are as defined above; more specifically, Ar 2is phenyl or 2-naphthyl;

[0073] R 4 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxyl, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 wherein Q 1 、Q 2 and R 9 are as defined above; specifically, R 4 is C1-C6 alkyl or C1-C6 alkoxy; more specifically, R 4 is C1-C2 alkyl or C1-C2 alkoxy; and

[0074] R 5 is hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl or C1-C4 alkanoyl; specifically, R 5 is hydrogen.

[0075] In one embodiment, the compounds of formula (I), (II), (III) described herein or their pharmaceutically acceptable salts are subject to the following conditions:

[0076] a) When Ar 1 is R 4 is -OMe, R 5 is hydrogen and Ar 2 is phenyl or phenyl substituted by R 8 for the compounds of formula (III), R 1 、R 2 and R 8 do not satisfy the following six conditions:

[0077]

[0078] b) When Ar 1 is R 1 is hydrogen, NO2, NH2, mono-C1-C8 alkylamino or di-C1-C8 alkylamino, R 2 is hydrogen or C1-C8 alkyl, R 4 is hydrogen or C1-C8 alkyl, and R 5 is hydrogen, then Ar 2is not an unsubstituted phenyl or a phenyl substituted with a C1-C8 alkyl group at the 2-position. As understood herein, the compounds excluded from formula (III) according to condition b) are also excluded from the scope of formula (I) and formula (II).

[0079] A compound of formula (I) or a pharmaceutically acceptable salt thereof,

[0080]

[0081] wherein Ar 1 、Ar 2 、R 4 and R 5 are as defined above.

[0082] A compound of formula (II) or a pharmaceutically acceptable salt thereof,

[0083]

[0084] wherein Ar 1 、Ar 2 、R 4 and R 5 are as defined above.

[0085] A compound of formula (III) or a pharmaceutically acceptable salt thereof,

[0086]

[0087] wherein Ar 1 、Ar 2 、R 4 and R 5 are as defined above.

[0088] In one embodiment, a compound of formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, wherein Ar 1 is R 1 and R 2 are as defined above.

[0089] In one embodiment, a compound of formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, wherein R 4 is a C1-C8 alkoxy group, including a branched alkoxy group; more specifically, R 4 is a C1-C4 alkoxy group.

[0090] In one embodiment, a compound of formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, wherein Ar 1 is R 1 and R2 As defined above, and R 4 is a C1-C8 alkoxy group, including branched alkoxy groups.

[0091] In one embodiment, a compound of formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, wherein Ar 2 is a phenyl group substituted at the 2-position by R 8 . In this embodiment, R 8 is a halogen, specifically F.

[0092] In one embodiment, a compound of formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, wherein Ar 1 is R 1 and R 2 are as defined above, and Ar 2 is a phenyl group substituted at the 2-position by R 8 ; specifically, R 8 is a halogen, specifically F. In a further embodiment, R 4 is a C1-C8 alkoxy group, including branched alkoxy groups.

[0093] In one embodiment, a compound of formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, wherein Ar 1 is R 1 is hydrogen, methyl, ethyl, methoxy, NH2, NO2, -CH2OH or -Q 1 (C═O)Q 2 R 9 , R 9 is hydrogen, a C1-C8 alkyl group, a C1-C6 haloalkyl group, a C1-C8 alkyl-OH or an aryl group, and Q 1 and Q 2 are NH; and R 2 is methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, -CH2OH, -CH2NH2 or phenyl; R 4 is a C1-C4 alkoxy group; R 5 is hydrogen; and Ar 2 is a phenyl group substituted at the 2-position by R 8 ; specifically, R 8 is a halogen, specifically F.

[0094] The present disclosure also includes non-salt forms (such as free base forms) of the compounds of formula (I) listed in Table 1 or pharmaceutically acceptable salts thereof.

[0095] These compounds are described using standard nomenclature. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. Unless the context clearly dictates otherwise, each compound name includes the free acid or free base form of the compound, the hydrate of the compound, and all pharmaceutically acceptable salts of the compound.

[0096] As used herein, the terms “Formula (I)”, “Formula (II)”, and “Formula (III)” encompass all compounds that satisfy Formulas (I), (II), and (III), including any enantiomers, racemates, and stereoisomers, as well as all pharmaceutically acceptable salts, solvates, and hydrates of these compounds.

[0097] In certain cases, the compounds of Formulas (I), (II), and (III) may contain one or more asymmetric elements, such as stereocenters, stereogenic axes, etc., for example, asymmetric carbon atoms, such that the compounds can exist in different stereoisomeric forms. These compounds can be, for example, racemates or optically active forms. For compounds having two or more asymmetric elements, these compounds can also be mixtures of diastereomers. For compounds having an asymmetric center, it should be understood that all optical isomers and their mixtures are encompassed. In these cases, a single enantiomer, i.e., an optically active form, can be obtained by asymmetric synthesis, synthesis from optically pure precursors, or by resolution of a racemate or racemic intermediate. Resolution of a racemate can also be achieved by conventional methods, for example, crystallization in the presence of a resolving agent, or chromatography using, for example, a chiral HPLC column.

[0098] When a compound exists in various tautomeric forms, the compound is not limited to any one particular tautomer, but includes all tautomeric forms.

[0099] All isotopes of atoms occurring in the compounds of the present invention are contemplated. Isotopes include those atoms having the same atomic number but different mass numbers. By way of general example (but not limitation), isotopes of hydrogen include tritium and deuterium, and isotopes of carbon include 11 C, 13 C, and 14 C.

[0100] As used herein, general formulas containing variables (such as R 1 -R 9 、Y, Z, etc.) are used to describe certain compounds. Unless otherwise specified, each variable in such general formulas is defined independently of the other variables. Thus, if a group is said to be, for example, substituted with 0 - 2 R 1 groups, then the group can be substituted with up to two R 1 groups, and each occurrence of R1 are each independently selected from the definition of R 1 Moreover, such a combination is permitted only if the combination of substituents and / or variables results in a stable compound.

[0101] As used herein, the term "active ingredient" refers to a compound (including compounds of formulas (I), (II) and (III)), element or mixture that directly or indirectly produces a physiological effect on a patient when administered to the patient alone or in combination with another compound, element or mixture. The indirect physiological effect can occur through metabolites or other indirect mechanisms. When the active ingredient is a compound, it includes salts, solvates (including hydrates), crystal forms, amorphous forms and any polymorphs of the free compound. All forms are contemplated herein regardless of the method used to obtain them.

[0102] A short dash ("-") not located between two letters or symbols is used to indicate the point of attachment of a substituent. For example, -(CH2)C3-C8 cycloalkyl is attached through the carbon of the methylene (CH2) group.

[0103] "Alkanoyl" is an alkyl as defined herein covalently bound to the group it replaces through a keto group (-(C=O)-) bridge. The alkanoyl has a specified number of carbon atoms, with the carbon of the keto group included in the numbered carbon atoms. For example, C2 alkanoyl is acetyl having the chemical formula CH3(C=O)-. Alkanoyl of C4 or more carbon atoms can include cycloalkyl (such as cyclopropyl) as well as straight-chain or branched-chain groups.

[0104] As used herein, the term "alkyl" refers to a branched or straight-chain saturated aliphatic hydrocarbon group having a specified number of carbon atoms (usually 1 to about 12 carbon atoms). As used herein, the term C1-C6 alkyl represents an alkyl having 1, 2, 3, 4, 5 or 6 carbon atoms. Other embodiments include alkyl having 1 to 8 carbon atoms, 1 to 4 carbon atoms or 1 or 2 carbon atoms, such as C1-C6 alkyl, C1-C4 alkyl and C1-C2 alkyl. When C0-C n alkyl is used in combination with another group, such as (cycloalkyl)C0-C4 alkyl, the specified group in this case (i.e., cycloalkyl) is either directly attached through a single covalent bond (C0) or through an alkyl chain having a specified number of carbon atoms (in this case 1, 2, 3 or 4 carbon atoms). Examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, 3-methylbutyl, tert-butyl, n-pentyl and sec-pentyl.

[0105] As used herein, the term "cycloalkyl" refers to a saturated hydrocarbon ring group having only carbon ring atoms and having a specified number of carbon atoms (usually 3 to about 8 ring carbon atoms, or 3 to about 7 carbon atoms). Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, as well as bridged or cage-like saturated ring groups such as norbornane or adamantane.

[0106] As used herein, the term "heterocycloalkyl" refers to a saturated cyclic group containing 1 to about 3 heteroatoms selected from N, O and S and the remaining ring atoms being carbon. Heterocycloalkyl has 3 to about 8 ring atoms, more typically 5 to 7 ring atoms. Examples of heterocycloalkyl include morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl. The nitrogen in heterocycloalkyl may optionally be quaternized.

[0107] As used herein, the term "alkenyl" refers to straight-chain and branched-chain hydrocarbon chains containing one or more unsaturated carbon-carbon bonds, which may occur at any stable point on the chain. The alkenyls described herein generally have 2 to about 12 carbon atoms. Exemplary alkenyls are lower alkenyls, i.e., those having 2 to about 8 carbon atoms, such as C2-C8, C2-C6 and C2-C4 alkenyls. Examples of alkenyl include vinyl, propenyl and butenyl.

[0108] As used herein, the term "cycloalkenyl" refers to a saturated hydrocarbon ring group containing one or more unsaturated carbon-carbon bonds (which may occur at any stable point on the ring) and having a specified number of carbon atoms. Monocyclic cycloalkenyls generally have 3 to about 8 carbon ring atoms or 3 to 7 (3, 4, 5, 6 or 7) carbon ring atoms. Cycloalkenyl substituents may dangle from the substituted nitrogen or carbon atom, or a substituted carbon atom having two substituents may have a cycloalkenyl attached in the form of a spiro group. Examples of cycloalkenyl include cyclopropenyl, cyclobutenyl, cyclopentenyl or cyclohexenyl, as well as bridged or cage-like saturated ring groups such as norbornene.

[0109] As used herein, the term "heterocycloalkenyl" refers to a 3- to 10-membered (including 4- to 8-membered) non-aromatic ring structure having one or more double bonds and one or more of the ring atoms being heteroatoms (such as N, O or S).

[0110] As used herein, the term "(cycloalkyl)C0-C n alkyl" refers to a substituent in which cycloalkyl and alkyl are as defined herein, and the point of attachment of the (cycloalkyl)alkyl group to the molecule it replaces is either a single covalent bond (C0 alkyl) or on the alkyl group. (Cycloalkyl)alkyl includes, but is not limited to, cyclopropylmethyl, cyclobutylmethyl and cyclohexylmethyl.

[0111] As used herein, the term "(heterocycloalkyl)C0-C n"Alkyl" refers to a substituent in which a heteroalkyl and an alkyl are as defined herein, and the point of attachment of the (heteroalkyl)alkyl group to the molecule it replaces is either a single covalent bond (C0 alkyl) or on the alkyl group. (Heteroalkyl)alkyl includes, but is not limited to, morpholinomethyl, piperazinomethyl, piperidinomethyl, and pyrrolidinomethyl.

[0112] As used herein, the term "(heteroalkenyl)C0-C6 alkyl" refers to a substituent in which a heteroalkenyl and an alkyl are as defined herein, and the point of attachment of the (heteroalkenyl)alkyl group to the molecule it replaces is either a single covalent bond (C0 alkyl) or on the alkyl group.

[0113] As used herein, the term "aryl" refers to an aromatic group containing only carbon in an aromatic ring or multiple aromatic rings. Typical aryls contain 1 to 3 independent, fused, or pendant rings and contain 6 to about 18 ring atoms with no heteroatoms among the ring members. When indicated, such aryls may be further substituted by carbon or non-carbon atoms or groups. Bicyclic aryls may be further substituted by carbon or non-carbon atoms or groups. Bicyclic aryls may contain two fused aromatic rings (naphthyl) or one aromatic ring fused to a 5- to 7-membered non-aromatic cyclic group, which non-aromatic cyclic group optionally contains 1 or 2 heteroatoms independently selected from N, O, and S, such as 3,4-methylenedioxyphenyl. Aryl includes, for example, phenyl, naphthyl (including 1-naphthyl and 2-naphthyl), and biphenyl.

[0114] As used herein, the term "monocyclic or bicyclic heteroaryl" refers to a stable 5- to 7-membered monocyclic heterocycle or 7- to 10-membered bicyclic heterocycle containing at least 1 aromatic ring containing 1 to 4 (or specifically 1 to 3) heteroatoms selected from N, O, and S and the remaining ring atoms being carbon. When the total number of S and O atoms in the heteroaryl exceeds 1, these heteroatoms are not adjacent to each other. Specifically, the total number of S and O atoms in the heteroaryl does not exceed 2, and more specifically, the total number of S and O atoms in the heteroaryl does not exceed 1. The nitrogen atoms in the heteroaryl may be optionally quaternized. When indicated, such heteroaryls may be further substituted by carbon or non-carbon atoms or groups. Such substitution may include fusion with a 5- to 7-membered saturated cyclic group, which saturated cyclic group optionally contains 1 or 2 heteroatoms independently selected from N, O, and S, to form, for example, [1,3]dioxoleno[4,5-c]pyridinyl. In certain embodiments, 5- to 6-membered heteroaryls are used. Examples of heteroaryls include, but are not limited to, pyridyl, indolyl, pyrimidinyl, pyridazinyl, pyrazinyl, imidazolyl, oxazolyl, furyl, thienyl, thiazolyl, triazolyl, tetrazolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl, benz[b]thienyl, isoquinolinyl, quinazolinyl, quinoxalinyl, thienyl, isoindolyl, and 5,6,7,8-tetrahydroisoquinoline.

[0115] "Halogenated alkyl" includes branched and straight-chain alkyl groups having a specified number of carbon atoms and substituted with one or more (up to the maximum allowable number of halogen atoms) halogen atoms. Examples of halogenated alkyls include, but are not limited to, trifluoromethyl, difluoromethyl, 2-fluoroethyl, and pentafluoroethyl.

[0116] "Alkoxy" is an alkyl group linked by an oxygen bridge (the oxygen of the alcohol group) as defined herein.

[0117] "Halogenated alkoxy" is a halogenated alkyl group linked by an oxygen bridge (the oxygen of the alcohol group) as defined herein.

[0118] "Halogenated" or "halogen" is any one of fluorine, chlorine, bromine, and iodine.

[0119] "Monoalkylamino and / or dialkylamino" is a secondary or tertiary alkylamino, wherein the alkyl is an independently selected alkyl having a specified number of carbon atoms as defined herein. The point of attachment of the alkylamino is on the nitrogen. Examples of monoalkylamino and dialkylamino include ethylamino, dimethylamino, and methyl-propyl-amino.

[0120] As used herein, the term "substituted" means that any one or more hydrogen atoms on a specified atom or group are replaced by a group selected from the specified group, provided that the normal valence of the specified atom is not exceeded. When the substituent is oxo (i.e., =O), two hydrogens on the atom are replaced. When the oxo group replaces an aromatic moiety, the corresponding partially unsaturated ring replaces the aromatic ring. For example, pyridyl substituted with oxo is pyridone. Only combinations of substituents and / or variables that result in a stable compound or useful synthetic intermediate are permitted. A stable compound or stable structure means a compound that is stable enough to be isolated from a reaction mixture and subsequently formulated into an effective therapeutic agent.

[0121] Unless otherwise specified, substituents are named into the core structure. For example, it should be understood that when (cycloalkyl)alkyl is listed as a possible substituent, the point of attachment of the substituent to the core structure is on the alkyl moiety; or when arylalkyl is listed as a possible substituent, the point of attachment to the core structure is the alkyl moiety.

[0122] Suitable groups that may be present in "substituted" or "optionally substituted" positions include, but are not limited to: halogen; cyano; hydroxy; nitro; azido; alkanoyl (e.g., C2-C6 alkanoyl, such as acyl, etc.); formamido; alkyl having 1 to about 8 carbon atoms, or 1 to about 6 carbon atoms (including cycloalkyl); alkenyl and alkynyl including groups having one or more unsaturated bonds and containing 2 to about 8 carbon atoms, or 2 to about 6 carbon atoms; alkoxy having one or more oxygen bonds and containing 1 to about 8 carbon atoms, or 1 to about 6 carbon atoms; aryloxy, such as phenoxy; alkylthionyl groups, including alkylthionyl groups having one or more thionyl bonds and containing 1 to about 8 carbon atoms, or 1 to about 6 carbon atoms; alkylsulfinyl groups, including alkylsulfinyl groups having one or more sulfinyl bonds and containing 1 to about 8 carbon atoms, or 1 to about 6 carbon atoms; alkylsulfonyl groups, including alkylsulfonyl groups having one or more sulfonyl bonds and containing 1 to about 8 carbon atoms, or 1 to about 6 carbon atoms; aminoalkyl groups, including aminoalkyl groups having one or more N atoms and containing 1 to about 8 carbon atoms, or 1 to about 6 carbon atoms; aryl having 6 or more carbon atoms and one or more rings (e.g., phenyl, biphenyl, naphthyl, etc., each ring being a substituted or unsubstituted aromatic ring); arylalkyl having 1 to 3 independent or fused rings and containing 6 to about 18 ring carbon atoms, an example of arylalkyl being benzyl; aryloxyalkyl having 1 to 3 independent or fused rings and containing 6 to about 18 ring carbon atoms, an example of aryloxyalkyl being benzyloxy; or a saturated, unsaturated or aromatic heterocyclic group having 1 to 3 independent or fused rings, each ring having 3 to about 8 members and containing one or more N, O or S atoms, such as coumarinyl, quinolinyl, isoquinolinyl, quinazolinyl, pyridyl, pyrazinyl, pyrimidinyl, furyl, pyrrolyl, thienyl, thiazolyl, triazinyl, oxazolyl, isoxazolyl, imidazolyl, indolyl, benzofuranyl, benzothiazolyl, tetrahydrofuryl, tetrahydropyranyl, piperidyl, morpholinyl, piperazinyl and pyrrolidinyl. These heterocyclic groups may be further substituted, for example, by hydroxy, alkyl, alkoxy, halogen and amino.

[0123] As used herein, the term "pharmaceutically acceptable salt" includes derivatives of the disclosed compounds in which the parent compound is modified by making acid addition salts, or base addition salts, of inorganic and organic acids. Salts of the compounds of the invention may be synthesized from the parent compound, which contains a basic or acidic moiety, by conventional chemical methods. Generally, such salts are prepared by reacting the free acid form of these compounds with a stoichiometric amount of the appropriate base such as hydroxides, carbonates, bicarbonates, etc. of Na, Ca, Mg or K, or by reacting the free base form of these compounds with a stoichiometric amount of the appropriate acid. Such reactions are generally carried out in water or an organic solvent, or in a mixture of both. Generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are used when feasible. Salts of the compounds of the invention also include solvates of the compounds and compound salts.

[0124] Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines; alkali metal or organic salts of acidic residues such as carboxylic acids, etc. Pharmaceutically acceptable salts include conventional non-toxic salts and quaternary ammonium salts formed from the parent compound, such as salts formed from non-toxic inorganic or organic acids. For example, conventional non-toxic acid salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, etc.; and salts formed from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, HOOC-(CH2) n -COOH (where n is 0 - 4), etc. Lists of other suitable salts can be found, for example, in Remington’s Pharmaceutical Sciences, 17th Edition, Mack Publishing Company, Easton, Pennsylvania, page 1418 (1985).

[0125] There are also provided pharmaceutical compositions comprising a compound of formula (I) - (III) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. Such pharmaceutical compositions may comprise a compound of formula (I) - (III) as the sole active ingredient, or may comprise a combination with additional pharmaceutically active ingredients, including additional extracellular thiol isomerase inhibitors or active ingredients from different drug classes, including antithrombotic and anticoagulant agents, anti-inflammatory agents, antiviral agents, chemotherapeutic agents or anticancer agents, etc., or combinations thereof.

[0126] As used herein, "extracellular thiol isomerase inhibitor compound" is an inhibitor of one or more extracellular thiol isomerases. Exemplary extracellular thiol isomerase inhibitor compounds include compounds of formula (I), (II), (III) or pharmaceutically acceptable salts thereof, zafirlukast, montelukast, quercetin, PACMA-31, curcumin, rutin, isoquercitrin, CCF642, and combinations thereof.

[0127] The compound can be administered orally, topically, parenterally, by inhalation or spray, sublingually, transdermally, buccally, rectally, as an eye drop, or by other means, and its dosage form contains a conventional pharmaceutically acceptable carrier. The pharmaceutical composition can be formulated into any pharmaceutically useful form, such as an aerosol, cream, gel, pill, capsule, tablet, syrup, transdermal patch, or eye drop. Some dosage forms, such as tablets and capsules, can be subdivided into appropriate-sized unit doses containing an appropriate amount of the active ingredient, e.g., an effective amount for the intended purpose.

[0128] As used herein, the term "dosage form" refers to a unit of administration of the active ingredient. Examples of dosage forms include tablets, capsules, injections, suspensions, liquids, emulsions, creams, ointments, suppositories, inhalable forms, transdermal forms, etc. An exemplary dosage form is a solid oral dosage form.

[0129] As used herein, the term "pharmaceutical composition" refers to a composition comprising at least one active ingredient (such as a compound or salt of formula (I), (II), or (III)) and at least one other substance (such as a carrier). The pharmaceutical composition complies with the Good Manufacturing Practice (GMP) standards of the U.S. Food and Drug Administration (U.S. FDA) for drugs for human or non-human use. The pharmaceutical composition can be formulated into a dosage form.

[0130] As used herein, the term "carrier" when applied to a pharmaceutical composition refers to a diluent, excipient, or vehicle provided together with the active compound.

[0131] Carriers include excipients and diluents and must have a purity high enough and a toxicity low enough to make them suitable for administration to the patient being treated. The carrier can be inert or can have a pharmaceutical benefit of its own. The amount of carrier used with the compound is sufficient to provide a practical amount of material for administering each unit dose of the active compound.

[0132] The classes of carriers include, for example, buffering agents, coloring agents, diluents, disintegrants, emulsifying agents, flavoring agents, glidants, lubricants, preservatives, stabilizers, surfactants, tabletting agents, and wetting agents. Some carriers may be listed in more than one class; for example, vegetable oils may be used as lubricants in some formulations and as diluents in other formulations. Exemplary pharmaceutically acceptable carriers include sugars, starches, celluloses, tragacanth powder, malt, gelatin, talc, and vegetable oils. The pharmaceutical compositions may optionally contain other active ingredients which do not substantially interfere with the activity of the compounds of formula (I), (II), (III), etc.

[0133] The pharmaceutical compositions may be formulated for oral administration. These compositions contain from 0.1 to 99 weight percent (“wt. %”) of the active compounds of formula (I), (II), (III) or their pharmaceutically acceptable salts, specifically at least about 5 wt. %. In some embodiments, the composition contains from about 25 wt. % to about 50 wt. % or from about 5 wt. % to about 75 wt. % of the active compound.

[0134] As used herein, the term “therapeutically effective amount” of a pharmaceutical composition means an amount effective to provide a therapeutic benefit (e.g., preventing or ameliorating symptoms) when administered to a patient, such as for treating a patient suffering from a disease or disorder affected by one or more extracellular thiol isomerase activities. The therapeutically effective amount may vary depending on factors such as the disease state, the age and weight of the patient, and the ability of the compound to elicit the desired response in the patient. The dosing regimen may be adjusted to provide the optimal therapeutic response. The therapeutically effective amount also means an amount where any toxic or detrimental effects (e.g., side effects) of the inhibitor compound are offset by the therapeutic beneficial effects.

[0135] The therapeutically effective amount of the compounds of formula (I), (II), (III) or their pharmaceutically acceptable salts may range from about 0.001 μg / kg / day to about 500 mg / kg / day, preferably from 0.01 μg / kg / day to 100 mg / kg / day. Those skilled in the art will understand that certain factors may affect the dosage required to effectively treat a patient, including but not limited to the severity of the disease or disorder, previous treatment, the overall health and / or age of the patient, and the presence of other diseases. In addition, treating a patient with a therapeutically effective amount of an inhibitor compound may include a single treatment, or may include a series of treatments. It will also be understood that the effective dose of the inhibitor compound used for treatment may increase or decrease during a particular course of treatment.

[0136] The compounds of formula (I), (II), (III) or their pharmaceutically acceptable salts may be administered to a patient in need thereof once, twice, or three times daily. In this example, the administration may be oral.

[0137] When administered orally, the total daily dose of the compounds of formula (I), (II), (III) or their pharmaceutically acceptable salts can be from about 0.1 to about 200 mg, specifically from about 1 to about 175 mg, more specifically from about 20 to about 150 mg, and even more specifically from about 60 to about 125 mg, administered orally once, twice or three times a day.

[0138] The pharmaceutical composition can be formulated into a package that contains the pharmaceutical composition in a container and also contains instructions for using the composition to prevent and treat diseases or disorders mediated by one or more thiol isomerases in extracellular thiol isomerase.

[0139] Also provided is a method of treatment using the compounds of formula (I), (II), (III) or their pharmaceutically acceptable salts. In one embodiment, a method of treating or preventing cancer-induced thrombosis, comprising administering to a patient in need a therapeutically effective amount of the compounds of formula (I), (II), (III) or their pharmaceutically acceptable salts, optionally in the form of a pharmaceutical composition comprising the compound.

[0140] In one embodiment, a method of inhibiting one or more extracellular thiol isomerases in a patient in need to treat or prevent a disease or disorder affected by the activity of one or more extracellular thiol isomerases, or inhibiting a process affected by the activity of one or more extracellular thiol isomerases, the method comprising administering to the patient a therapeutically effective amount of the compounds of formula (I), (II), (III) or their pharmaceutically acceptable salts or a composition comprising the compound. In certain embodiments, as described herein, the disease or disorder is thrombosis, cancer-induced thrombosis, thrombotic disease, infectious disease including human immunodeficiency virus (HIV), cancer, inflammation or a combination thereof.

[0141] Extracellular thiol isomerase

[0142] As used herein, "extracellular thiol isomerase" includes at least protein disulfide isomerase (PDI), thioredoxin (TRX) and the following endoplasmic reticulum resident proteins: ERp5, ERp57 and ERp72.

[0143] PDI is a 508 amino acid protein and plays an important role in platelet activation. It was confirmed nearly two decades ago that activated platelets release PDI, and this has been confirmed by many subsequent studies. Endothelial cells also release PDI. Antibodies against PDI and micromolar concentrations of the antibiotic bacitracin can inhibit platelet aggregation, adhesion and secretion. In an ovarian cancer model, inhibition of PDI has been shown to inhibit cancer cell growth and induce tumor necrosis.

[0144] ERp5 is a 440 - amino - acid protein that contains two thioredoxin (CGHC - containing motif) domains and has 47% sequence identity with PDI. Blocking ERp5 on the cell surface leads to reduced platelet aggregation, fibrinogen binding, and α - granule secretion. In addition to its role in hemostasis, high levels of ERp5 expression have been shown to be associated with preventing an effective anti - tumor response in Hodgkin lymphoma and have been proposed as a biomarker for prostate and breast cancer progression.

[0145] ERp57 is a 505 - amino - acid thiol isomerase that plays an important role in regulating initial platelet activation and supporting arterial thrombosis, affecting platelet aggregation, dense granule secretion, fibrinogen binding, calcium mobilization, and thrombus formation under arterial flow conditions. In addition to its role in thrombosis, ERp57 has also been shown to be required for the correct folding of influenza hemagglutinin and is associated with the disease progression of Alzheimer’s disease and cancer metastasis.

[0146] ERp72 is a 645 - amino - acid soluble endoplasmic reticulum (ER) protein that has 37% sequence homology with PDI. ERp72 has three catalytic CGHC domains, while PDI, ERp5, and ERp57 have only two catalytic CGHC domains. The percentage increase in ERp72 recruited to the platelet surface after activation is higher than that of PDI, ERp5, and ERp57, indicating that it plays an important but currently unknown role in platelet activation. The effect of ERp72 inhibition on thrombosis is also unknown. In addition to being re - localized to the surface of activated platelets, ERp72 is also associated with the infection process of polyomavirus and the redox signaling of NADPH oxidase (Nox) 1.

[0147] Four thiol isomerases, PDI, ERp5, ERp57, and ERp72, have been shown to be important for cancer - associated thrombosis. These four enzymes are non - redundant, i.e., inhibiting only one of them can prevent thrombus formation. Therefore, one or two or a combination of more of these enzymes are attractive targets for anti - thrombus drug development. In addition, some cancers are dependent on these enzymes, which would make inhibitors of these enzymes have anti - cancer activity in these cases.

[0148] Zafirlukast and montelukast are inhibitors of the four thiol isomerases PDI, ERp5, ERp57, and ERp72, which can inhibit both venous and arterial thrombosis without increasing the risk of bleeding or promoting cancer cell and tumor growth. Certain zafirlukast derivatives described herein show increased activity compared with zafirlukast.

[0149] In one embodiment, the inhibition target is one or more thiol isomerases in extracellular thiol isomerase, including protein disulfide isomerase (PDI), thioredoxin (TRX), ERp5, ERp57, and ERp72. In one embodiment, the inhibition target is PDI. In one embodiment, the inhibition target is thioredoxin (TRX). In one embodiment, the inhibition target is ERp5. In one embodiment, the inhibition target is ERp57. In one embodiment, the inhibition target is ERp72.

[0150] As used herein, the term "patient" refers to a human or non-human animal in need of medical treatment. Medical treatment can include treatment of an existing condition (such as a disease or disorder), preventive or prophylactic treatment, or diagnostic treatment. In some embodiments, the patient is a human patient.

[0151] As used herein, the term "provide" means to give, administer, sell, distribute, transfer (whether for profit or not), manufacture, compound, or dispense.

[0152] As used herein, the term "providing a compound of formula (I), (II), (III) or a pharmaceutically acceptable salt thereof and an additional pharmaceutically active ingredient" means that the compound of formula (I), (II), (III) or a pharmaceutically acceptable salt thereof and an additional pharmaceutically active ingredient are provided simultaneously in a single dosage form, provided simultaneously in separate dosage forms, or provided in separate dosage forms and administered at separate time intervals, the time interval being the time when the compound of formula (I), (II), (III) or a pharmaceutically acceptable salt thereof and an additional pharmaceutically active ingredient are present simultaneously in the patient's bloodstream. The compound of formula (I), (II), (III) or a pharmaceutically acceptable salt thereof and an additional pharmaceutically active ingredient do not have to be prescribed for the patient by the same healthcare provider. The additional pharmaceutically active ingredient does not require a prescription. Administration of the compound of formula (I), (II), (III) or a pharmaceutically acceptable salt thereof or the additional pharmaceutically active ingredient can be carried out by any suitable route, such as oral tablets, oral capsules, oral liquids, inhalation, injection, suppositories, or topical application.

[0153] As used herein, the term "providing an extracellular thiol isomerase inhibitor compound or a pharmaceutically acceptable salt thereof and at least one additional therapeutic agent" means that the extracellular thiol isomerase inhibitor compound or a pharmaceutically acceptable salt thereof and the additional active ingredient are provided simultaneously in a single dosage form, provided simultaneously in separate dosage forms, or provided in separate dosage forms and administered separately at intervals of time that result in the extracellular thiol isomerase inhibitor compound or a pharmaceutically acceptable salt thereof and at least one additional active ingredient being present simultaneously in the bloodstream of the patient. The extracellular thiol isomerase inhibitor compound or a pharmaceutically acceptable salt thereof and the additional active ingredient need not be prescribed for the patient by the same healthcare provider. The additional active ingredient does not require a prescription. Administration of the extracellular thiol isomerase inhibitor compound or a pharmaceutically acceptable salt thereof or at least one additional active ingredient can be by any suitable route, such as oral tablets, oral capsules, oral liquids, inhalation, injection, suppositories, or topical application.

[0154] As used herein, the term "treating" includes providing an extracellular thiol isomerase inhibitor compound, a compound of formula (I), (II), (III), or a pharmaceutically acceptable salt thereof, as the sole active ingredient or together with at least one additional active ingredient, sufficient to: (a) prevent a disease or disorder, or the symptoms of a disease or disorder, from occurring in a patient who may be predisposed to the disease or disorder but has not been diagnosed as having the disease or disorder; (b) inhibit a disease or disorder, i.e., prevent its progression; and (c) alleviate a disease or disorder, i.e., cause regression of the disease or disorder. "Treating / treatment" also refers to providing a therapeutically effective amount of an extracellular thiol isomerase inhibitor compound or a pharmaceutically acceptable salt thereof, as the sole active ingredient or together with at least one additional active ingredient, to a patient suffering from a disease or disorder affected by one or more extracellular thiol isomerase activities. "A disease or disorder affected by one or more extracellular thiol isomerase activities" means that the one or more extracellular thiol isomerases are involved with the disease or disorder.

[0155] The extracellular thiol isomerase inhibitor compounds or pharmaceutical compositions / combinations disclosed herein can be used to treat a patient. The extracellular thiol isomerase inhibitor compounds or pharmaceutical compositions / combinations can be used to treat or prevent diseases and disorders involving the activity of one or more extracellular thiol isomerases. In certain embodiments, the patient has thrombosis or is at risk of developing thrombosis. In certain embodiments, the patient has cancer. In certain embodiments, the patient is at risk of or has cancer-induced thrombosis. In certain embodiments, the disease is a hematological cancer, an HPV-related cancer, ovarian cancer, prostate cancer, gastric cancer, breast cancer, or colorectal cancer. In other embodiments, the patient to be treated has an inflammatory disease, an infectious disease, an immune disease, or a neurological disease.

[0156] Antithrombotic

[0157] Extracellular thiol isomerase is involved in the regulation of hemostasis and thrombosis because inhibition of one or more extracellular thiol isomerases will prevent platelet aggregation, granule secretion, adhesion, thrombosis, and fibrin formation. Antithrombotic drugs can be used for therapeutic prevention (primary prevention, secondary prevention) or treatment of dangerous blood clots (acute thrombosis).

[0158] In a mouse model of thrombosis, inhibition of the activity of PDI, ERp5, or ERp57 can prevent thrombosis after laser-induced vascular injury.

[0159] Furthermore, since inhibition of the activity of thiol isomerase affects platelet aggregation and fibrin formation, these treatment methods would be an improvement over existing therapies that currently target only arterial blood clots (heart attacks and strokes mainly caused by inappropriate activation of platelets) or venous blood clots (deep vein thrombosis and pulmonary embolism mainly caused by inappropriate activation of the coagulation system). Data indicate that each thiol isomerase has unique substrate specificity and mechanism of action, suggesting that each thiol isomerase can be an independent target.

[0160] The thrombotic diseases or disorders prevented or treated by the extracellular thiol isomerase inhibitor compounds can be acute myocardial infarction, stable angina, unstable angina, acute occlusion after coronary angioplasty and / or stent placement, transient ischemic attack, cerebrovascular disease, stroke, peripheral vascular disease, placental insufficiency, atrial fibrillation, deep vein thrombosis, pulmonary embolism, or a combination thereof.

[0161] Platelet responses in the presence of zileuton (0.1 μM - 10 μM) were tested by a series of platelet function assays, including aggregation, granule secretion, and spreading studies. Zileuton was found to inhibit platelet aggregation, dense granule and α-granule secretion, platelet spreading on collagen, and thrombus formation under flow conditions. These data suggest that zileuton and other broad-spectrum thiol isomerase inhibitors of the protein disulfide isomerase subfamily can be used as antithrombotic drugs.

[0162] In one embodiment, a disease or disorder affected by one or more extracellular thiol isomerase activities is arterial thrombosis, venous thrombosis, thrombotic diseases (such as acute myocardial infarction, stable angina, unstable angina, acute occlusion after coronary angioplasty and / or stent placement, transient ischemic attack, cerebrovascular disease, stroke, peripheral vascular disease, placental insufficiency, atrial fibrillation, deep vein thrombosis, and pulmonary embolism) or a combination thereof; and wherein the extracellular thiol isomerase inhibitor compound is a compound of formula (I), (II), (III) or a pharmaceutically acceptable salt thereof.

[0163] Anticancer

[0164] PDI inhibition is a viable target for cancer therapy. See Xu et al., "Protein Disulfide Isomerase: A Promising Target for Cancer Therapy," Drug Discovery Today, Vol. 19, No. 3, March 2014.

[0165] There is further evidence that ERp5 plays a role in tumor metastasis. See Gumireddy et al., "Screening for Metastasis-Promoting Genes in Mice," Proceedings of the National Academy of Sciences of the United States of America (PNAS) (2007) 104, pp. 6696 - 6701.

[0166] ERp57 is expressed by the human PDIA3 gene. ERp57 consists of 505 amino acids and is upregulated in breast, lung, uterine, gastric, and liver cancers, as well as melanoma, compared to normal tissues. The expression level of ERp57 is positively correlated with the transforming ability of the oncogenic sarcoma virus in NIH3T3 cells, indicating that ERp57 is involved in oncogenic transformation. Different from other PDI family members, ERp57 can interact with nuclear DNA and activate gene expression because ERp57 interacts with DNA molecules through its catalytically active a’ domain. ERp57 is also a component of the signal transducer and activator of transcription 3 (STAT3)-transcription complex, and ERp57-STAT3 regulates cell signaling and proliferation regulated by STAT3. ERp57 also regulates gene expression through the mammalian target of rapamycin (mTOR) pathway, which is another important regulator of cell proliferation and survival. ERp57 is also involved in binding to at least three proteins involved in DNA repair, including redox factor-1 / apurinic / apyrimidinic endonuclease (Ref-1 / APE), which itself has the ability to activate other transcription factors. In addition to its gene regulatory functions, increased ERp57 expression is associated with resistance to paclitaxel treatment in ovarian cancer and radioresistance in laryngeal cancer, promotes breast cancer metastasis to bone, and is involved in the dysregulation of epidermal growth factor receptor (EGFR) signaling in breast cancer cell lines, preventing the activation of its downstream target molecules (such as STAT3, Akt, and PLCγ). Since ERp57 plays a role in thrombosis, it is a potential target for preventing cancer-related thrombosis, which is a major cause of morbidity and mortality in cancer patients.

[0167] The cancer to be treated can be ovarian cancer, prostate cancer, lung cancer, melanoma, lymphoma, glioma, breast cancer, colon cancer, colorectal cancer, hematological cancer, laryngeal cancer, melanoma, or neuroblastoma.

[0168] The compounds of formula (I), (II), (III) or their pharmaceutically acceptable salts can be used alone or optionally in combination with another anticancer or chemotherapeutic drug.

[0169] In one embodiment, cancer is treated with a combination of a compound of formula (I), (II), (III) or its pharmaceutically acceptable salt and a chemotherapeutic drug (such as carboplatin or cisplatin).

[0170] The present invention is further illustrated by the following examples, which should not be construed as limiting.

[0171] Examples:

[0172] Example 1. Representative Scheme for Synthesizing Compounds

[0173] Derivatives of zileuton (ZAF) with skeletons I (indole), IIa, and IIb were prepared (Table 1). The arrangement of substituents on the benzoyl group (ring B) in skeleton I is the same as that in the parent ZAF, while the arrangement of substituents on ring B in skeletons IIa and IIb is different from that in ZAF. Using three different ZAF skeletons provided an ideal opportunity for preliminary structure-activity relationship (SAR) studies.

[0174] Scheme I.

[0175]

[0176] a. Et2SiH, TFA, CH2Cl2; b. KOH, MeOH / THF / H2O; c. arylsulfonamide, EDC·HCl, DMAP, CH2Cl2.

[0177] Scheme II.

[0178]

[0179] X = R 6 , O, S; Y = CH, N; R 11 = I, Br, Li; R 12 = MgI, MgBr

[0180] a. NIS, TFA, THF; b. NIH, CH2Cl2; c. NBS, Al2O3, Et3N, THF, hexane; d. NBS, DMF; e. n-BuLi, THF, hexane; f. I2, hexane, Et2O, H2O; g. Mg, I2, THF.

[0181]

[0182] X = R 6 , O, S; Y = CH, N; R 13 = I, Br, Li, MgI, MgBr

[0183] h. NaBH4, THF, i. PBr3, Et2O; j. n-BuLi, THF; k. n-BuLi, ZnBr2, Pd(PPh3)4, THF; l. n-BuLi, Et2O; m. Zn, I2, S-Phos, Pd2dba3, DMF; n. Zn, P(Ph)3, Pd(OAc)2, TMSCl, 1,2-dibromoethane, THF, o. Zn, Cl2Pd(PPh3)2, DMA, benzene; p. THF; q. KOH, MeOH / THF / H2O; r. arylsulfonamide, EDC·HCl, DMAP, CH2Cl2.

[0184] Scheme I is a general three-step linear synthesis method that will be used to prepare the proposed indole derivatives (alternative synthetic routes or additional steps may be used for the preparation of some of the proposed derivatives). This representative synthetic route shows: (a) a condensation reaction between the substituted indole and methyl 5-formyl-2-methoxybenzoate using triethylsilane, trifluoroacetic acid, and dichloromethane as the source of acid anhydride; (b) hydrolysis of the ester to the carboxylic acid using sodium hydroxide, methanol, tetrahydrofuran, and water; and then (c) amide coupling using arylsulfonamide, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (reductant), 4-dimethylaminopyridine, and dichloromethane.

[0185] Scheme II is a general synthetic method that will be used to prepare other heteroaryl and heterocyclic derivatives (containing N, O, or S) other than the indole ring. For Scheme II, the step that is different from Scheme I is the first step of the condensation reaction (labeled as step (a) in Scheme II), while steps (q) and (r) will be the same as steps (b) and (c) in Scheme I in terms of hydrolysis of the ester to the carboxylic acid and amide coupling (as the last two steps of the synthesis). For Scheme II, 5-membered and 6-membered heterocycles will first be iodinated, brominated, or lithium-added at the C-2 position of the 5-membered ring and the C-3 position of the 6-membered ring (using one of steps a-f). These compounds will either go directly to the next step (when R1 = iodine, bromine, or lithium) or undergo step (g) to synthesize a Grignard reagent to further drive the reaction forward (when R1 = magnesium iodide or magnesium bromide). This synthesis is a convergent synthesis, in which the aldehyde group of methyl 5-formyl-2-methoxybenzoate is reduced (h) to a primary alcohol, followed by (i) bromination, and then addition of a heterocycle with R1 = iodine, bromine, lithium, magnesium iodide, or magnesium bromide (using one of steps j-p) to form an ester, which undergoes the same reaction of hydrolysis of the ester to the carboxylic acid as step b in Scheme I, and the last step is amide coupling (step c in Scheme I). These steps make the 5-membered and 6-membered heterocycles more reactive to drive the reaction to completion and further prepare the final compound.

[0186] Representative compounds of formula (I), (II) or (III) are described in Table 1.

[0187] Table 1.

[0188]

[0189]

[0190]

[0191] *Based on turbidity assay of insulin; "ND"=>100 μM.

[0192] Materials and Apparatus. All chemicals were purchased from Oakwood Chemical (San Diego, CA), Combi-Blocks (San Diego, CA), TCI America (Portland, OR), Sigma Aldrich (St. Louis, MO), Synthonix (Wake Forest, NC), Matrix Scientific (Columbia, SC), Ricca Chemical (Arlington, TX), Acros Organics (Geel, Belgium), Beantown Chemical (Hudson, NH), Chem-Impex (Wooddale, IL), Alfa Aesar (Woldsea, MA) and used without further purification. All chemical reactions were performed using Merck silica gel 60F coated 254 The glass plates were monitored by thin layer chromatography (TLC). Ultraviolet light was used to visualize the bands on the TLC plates. Compounds were purified by silica gel column chromatography using F60 (40-63 μM, SiliCycle, Quebec, Canada). Spectra were recorded at 500 or 400 MHz using a Varian 500 (VNMRS500) or 400 (MR400) MHz spectrometer, respectively. 1 H NMR spectra. All were recorded at 100 MHz using a Varian 400 MHz spectrometer. 13 C NMR spectra. Chemical shifts (δ) for all NMR spectra are given in parts per million (ppm). All coupling constants (J) are given in Hertz (Hz), and the abbreviations for signal shapes are singlet (s), doublet (d), triplet (t), multiplet (m), double doublet (dd), double triplet (dt) and triple doublet (td). High resolution mass spectrometry (HRMS) was performed on an AB SCIEX TripleTOFTM5600 mass spectrometer.

[0193] Synthesis of Compound SM2. A solution of 5-nitroindole (3.0 g, 18.5 mmol) in anhydrous DMF (15 mL) was cooled to 0 °C and treated with NaH (60% in mineral oil, 1.48 g, 37.0 mmol). The reaction mixture was stirred at room temperature for 1 h. The mixture was cooled to 0 °C, then iodoethane (2.98 mL, 37.0 mmol) was added dropwise, and the resulting mixture was stirred at room temperature for 1 h. The reaction was quenched by pouring onto ice and extracted with CH2Cl2 (3×). The combined organic layers were washed with NaHCO3, H2O and brine, dried over MgSO4, filtered and concentrated. The resulting crude product was purified by column chromatography (SiO2 silica gel, hexane:EtOAc / 3:1, R f 0.46) to give Compound SM2 as a yellow solid (1.70 g, 48%): 1 H NMR (500 MHz, CDCl3) δ 8.58 (dd, J1 = 2.2 Hz, J2 = 0.5 Hz, 1H, aromatic), 8.10 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.34 (dt, J1 = 9.1 Hz, J2 = 0.7 Hz, 1H, aromatic), 7.25 (d, J = 3.3 Hz, 1H, aromatic), 6.66 (dd, J1 = 3.3 Hz, J2 = 0.9 Hz, 1H, aromatic), 4.21 (q, J = 7.4 Hz, 2H, NC H 2CH3), 1.49 (t, J = 7.4 Hz, 3H, NCH2C H 3); 13 C NMR (100 MHz, CDCl3) δ 138.7, 131.9, 130.4, 128.0, 118.5, 117.3, 109.3, 104.2, 41.7, 15.6.

[0194] Synthesis of Compound SM3. A solution of 5-nitroindole (3.0 g, 18.5 mmol) in anhydrous DMF (15 mL) was cooled to 0 °C and treated with NaH (60% in mineral oil, 1.48 g, 37.0 mmol). The reaction mixture was stirred at room temperature for 1 h. The mixture was cooled to 0 °C, then 1-iodopropane (3.59 mL, 37.0 mmol) was added slowly, and the resulting mixture was stirred at room temperature for 1 h. The reaction was quenched by pouring onto ice and extracted with CH2Cl2 (3×). The combined organic layers were washed with NaHCO3, H2O, and brine, dried over MgSO4, filtered, and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f = 0.81) to give Compound SM3 (2.71 g, 72%) as a brown liquid: 1 H NMR (500 MHz, CDCl3) δ 8.57 (d, J = 2.3 Hz, 1H, aromatic), 8.09 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.33 (dd, J1 = 9.1 Hz, J2 = 0.7 Hz, 1H, aromatic), 7.23 (d, J = 3.2 Hz, 1H, aromatic), 6.66 (dt, J1 = 3.3 Hz, J2 = 0.8 Hz, 1H, aromatic), 4.12 (t, J = 7.1 Hz, 2H, NC H 2CH2CH3), 1.88 (sextet, J = 7.4 Hz, 2H, NCH2C H 2CH3), 0.93 (t, J = 7.4 Hz, 3H, NCH2CH2C H 3); 13 C NMR (100 MHz, CDCl3) δ 141.6, 139.0, 131.2, 127.8, 118.4, 117.2, 109.4, 104.0, 48.7, 23.7, 11.6.

[0195] Synthesis of compound SM4KCH-3-26 (SGT1631). A solution of 5-nitroindole (3.0 g, 18.5 mmol) in anhydrous DMF (15 mL) was cooled to 0 °C and treated with NaH (60% in mineral oil, 1.48 g, 37.0 mmol). The reaction mixture was stirred at room temperature for 1 h. The mixture was cooled to 0 °C, then 1-iodobutane (4.20 mL, 37.0 mmol) was added dropwise, and the resulting mixture was stirred at room temperature for 1 h. The reaction was quenched by pouring onto ice and extracted with CH2Cl2 (3×). The combined organic layers were washed with NaHCO3, H2O and brine, dried over MgSO4, filtered and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 3:1, R f 0.53) to give compound SM4 (1.54 g, 38%) as a brown liquid: 1 H NMR (500 MHz, CDCl3) δ 8.57 (d, J = 2.2 Hz, 1H, aromatic), 8.09 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.33 (d, J = 9.1 Hz, 1H, aromatic), 7.23 (d, J = 3.3 Hz, 1H, aromatic), 6.65 (dd, J1 = 3.3 Hz, J2 = 0.8 Hz, 1H, aromatic), 4.15 (t, J = 7.2 Hz, 2H, NC H 2CH2CH2CH3), 1.82 (p, J = 7.7 Hz, 2H, NCH2C H 2CH2CH3), 1.33 (sextet, J = 7.7 Hz, 2H, NCH2CH2C H 2CH3), 0.93 (t, J = 7.4 Hz, 3H, NCH2CH2CH2C H 3); 13 C NMR (100 MHz, CDCl3) δ 141.7, 139.0, 131.2, 127.9, 118.5, 117.3, 109.4, 104.0, 46.8, 32.5, 20.3, 13.8.

[0196] Synthesis of compound SM5. A solution of 5-nitroindole (3.0 g, 18.5 mmol) in anhydrous DMF (15 mL) was cooled to 0 °C and treated with NaH (60% in mineral oil, 1.48 g, 37.0 mmol). The reaction mixture was stirred at room temperature for 1 h. The mixture was cooled to 0 °C, then 1-iodo-2-methylpropane (4.25 mL, 37.0 mmol) was slowly added, and the resulting mixture was stirred at room temperature for 1 h. The reaction was quenched by pouring onto ice and extracted with CH2Cl2 (3×). The combined organic layers were washed with NaHCO3, H2O and brine, dried over MgSO4, filtered and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.89) to give compound SM5 (1.12 g, 28%) as a yellow solid: 1 H NMR (500 MHz, CDCl3,) δ 8.57 (d, J = 2.3 Hz, 1H, aromatic), 8.09 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.32 (dt, J1 = 9.1 Hz, J2 = 0.7 Hz, 1H, aromatic), 7.20 (d, J = 3.3 Hz, 1H, aromatic), 6.66 (dd, J1 = 3.2 Hz, J2 = 0.8 Hz, 1H, aromatic), 3.94 (d, J = 7.4 Hz, 2H, NC H 2), 2.18 (septet, J = 7.3 Hz, 1H, C H (CH3)2), 0.92 (d, J = 6.7 Hz, 6H, CH(C H 3)2); 13 C NMR (100 MHz, CDCl3) δ 141.6, 139.3, 131.7, 127.8, 118.4, 117.3, 109.6, 103.9, 54.7, 29.9, 20.4 (2CH3).

[0197] Synthesis of compound SM6. A solution of 5-nitroindole (3.0 g, 18.5 mmol) in anhydrous DMF (15 mL) was cooled to 0 °C and treated with NaH (60% in mineral oil, 1.11 g, 27.8 mmol). The reaction mixture was stirred at room temperature for 1 h. The mixture was cooled to 0 °C, then benzyl bromide (3.30 mL, 27.8 mmol) was added dropwise, and the resulting mixture was stirred at room temperature for 1 h. The reaction was quenched by pouring onto ice and extracted with CH2Cl2 (3×). The combined organic layers were washed with NaHCO3, H2O and brine, dried over MgSO4, filtered and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 3:1, R f 0.58) to give compound SM6 (3.10 g, 66%) as a tan solid: 1 H NMR (500 MHz, CDCl3) δ 8.59 (dd, J1 = 2.3 Hz, J2 = 0.6 Hz, 1H, aromatic), 8.06 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.33 - 7.27 (m, 4H, aromatic), 7.26 (d, J = 3.3 Hz, 1H, aromatic), 7.09 - 7.18 (m, 2H, aromatic), 6.72 (dd, J1 = 3.3 Hz, J2 = 0.9 Hz, 1H, aromatic), 5.35 (s, 2H, NC H 2); 13 C NMR (100 MHz, CDCl3) δ 142.0, 139.2, 136.4, 131.7, 129.2 (2CH), 128.3, 128.1, 127.0 (2CH), 118.4, 117.6, 109.8, 104.6, 50.8.

[0198] Synthesis of compound SM9. A solution of compound SM2 (0.20 g, 1.05 mmol) and methyl 5-formyl-2-methoxybenzoate (0.20 g, 1.05 mmol) in anhydrous CH2Cl2 (5 mL) was cooled to 0 °C in an ice-water bath. Then Et3SiH (0.5 mL, 2.94 mmol) was added, followed by TFA (0.2 mL, 2.10 mmol). The mixture was stirred at 0 °C for 10 min and allowed to warm to room temperature overnight. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with NaHCO3, H2O and brine, dried over MgSO4, filtered and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f0.60) Purification gave compound SM9 as a yellow solid (0.24 g, 63%): 1 H NMR (500 MHz, CDCl3) δ 8.46 (dd, J1 = 2.2 Hz, J2 = 0.6 Hz, 1H, aromatic), 8.09 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.69 (d, J = 2.5 Hz, 1H, aromatic), 7.35 (dd, J1 = 8.6 Hz, J2 = 2.4 Hz, 1H, aromatic), 7.30 (dd, J1 = 9.2 Hz, J2 = 0.7 Hz, 1H, aromatic), 6.91 (d, J = 8.7 Hz, 1H, aromatic), 6.90 (s, 1H, aromatic), 4.14 (q, J = 7.4 Hz, 2H, NC H 2CH3), 4.06 (s, 2H, C H 2Ar), 3.87 (s, 3H, ArOC H 3), 3.85 (s, 3H, ArCO2C H 3), 1.44 (t, J = 7.4 Hz, 3H, NCH2C H 3); 13 C NMR (100 MHz, CDCl3) δ 167.0, 157.9, 141.4, 139.2, 133.8, 132.1, 131.9, 128.6, 127.3, 120.3, 117.7, 117.6, 116.8, 112.5, 109.3, 56.4, 52.3, 41.6, 30.4, 15.6.

[0199] Synthesis of compound SM10. A solution of compound SM3 (0.20 g, 0.98 mmol) and methyl 5-formyl-2-methoxybenzoate (0.19 g, 0.98 mmol) in anhydrous CH2Cl2 (5 mL) was cooled to 0 °C in an ice-water bath. Then Et3SiH (0.4 mL, 2.74 mmol) was added, followed by TFA (0.1 mL, 1.96 mmol). The mixture was stirred at 0 °C for 10 minutes and allowed to warm to room temperature overnight. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with NaHCO3, H2O and brine, dried over MgSO4, filtered and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.73) to give compound SM10 as a dark yellow solid (0.24 g, 63%): 11H NMR (500 MHz, CDCl3) δ 8.46 (dd, J1 = 2.3 Hz, J2 = 0.5 Hz, 1H, aromatic), 8.08 (dd, J1 = 9.1 Hz, J2 = 2.2 Hz, 1H, aromatic), 7.68 (d, J = 2.5 Hz, 1H, aromatic), 7.34 (dd, J1 = 8.6 Hz, J2 = 2.5 Hz, 1H, aromatic), 7.29 (d, J = 9.1 Hz, 1H, aromatic), 6.91 (d, J = 8.6 Hz, 1H, aromatic), 6.88 (app.t, J = 1.1 Hz, 1H, aromatic), 4.06 (s, 2H, C H 2Ar), 4.04 (t, J = 7.2 Hz, 2H, NC H 2CH2CH3), 3.87 (s, 3H, ArOC H 3), 3.85 (s, 3H, ArCO2C H 3), 1.83 (sextet, J = 7.4 Hz, 3H, NCH2C H 2CH3), 0.90 (t, J = 7.4 Hz, 3H, NCH2CH2C H 3); 13 13C NMR (100 MHz, CDCl3) δ 166.9, 157.9, 141.3, 139.6, 133.7, 132.1, 131.9, 129.4, 127.2, 120.2, 117.54, 117.46, 116.8, 112.5, 109.5, 56.3, 52.2, 48.5, 30.3, 23.8, 11.6.

[0200] Synthesis of compound SM11. A solution of compound SM4 (0.70 g, 3.21 mmol) and methyl 5-formyl-2-methoxybenzoate (0.62 g, 3.21 mmol) in anhydrous CH2Cl2 (5 mL) was cooled to 0 °C in an ice-water bath. Then Et3SiH (1.4 mL, 8.98 mmol) was added, followed by TFA (0.5 mL, 6.41 mmol). The mixture was stirred at 0 °C for 10 minutes and allowed to warm to room temperature overnight. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with NaHCO3, H2O, and brine, dried over MgSO4, filtered, and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.60) to give compound SM11 (0.10 g, 8%) as a yellow liquid (containing some methyl 5-formyl-2-methoxybenzoate, which was removed in the next synthetic step): 11H NMR (500 MHz, CDCl3) δ 8.45 (dd, J1 = 2.3 Hz, J2 = 0.5 Hz, 1H, aromatic), 8.08 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.68 (d, J = 2.4 Hz, 1H, aromatic), 7.34 (dd, J1 = 8.6 Hz, J2 = 2.5 Hz, 1H, aromatic), 7.29 (d, J = 9.1 Hz, 1H, aromatic), 6.90 (d, J = 8.6 Hz, 1H, aromatic), 6.88 (app.t, J = 1.1 Hz, 1H, aromatic), 4.07 (t, J = 7.3 Hz, 2H, NC H 2CH2CH2CH3), 4.05 (s, 2H, C H 2Ar), 3.87 (s, 3H, ArOC H 3), 3.85 (s, 3H, ArCO2C H 3), 1.81 - 1.75 (m, 2H, NCH2C H 2CH2CH3), 1.34 - 1.27 (m, 2H, NCH2CH2C H 2CH3), 0.92 (t, J = 7.4 Hz, 3H, NCH2CH2CH2C H 3); 13 13C NMR (100 MHz, (CD3)2SO) δ 166.2, 156.5, 140.2, 134.8, 133.3, 132.5, 130.4, 126.4, 119.8, 116.54, 116.47, 116.0, 113.2, 112.7, 110.4, 55.8, 51.8, 31.9, 29.0, 19.4, 13.5.

[0201] Synthesis of compound SM12. A solution of compound SM5 (0.20 g, 0.92 mmol) and methyl 5 - formyl - 2 - methoxybenzoate (0.18 g, 0.92 mmol) in anhydrous CH2Cl2 (5 mL) was cooled to 0 °C in an ice - water bath. Then Et3SiH (0.4 mL, 2.57 mmol) was added, followed by TFA (0.1 mL, 1.83 mmol). The mixture was stirred at 0 °C for 10 minutes and allowed to warm to room temperature overnight. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with NaHCO3, H2O, and brine, dried over MgSO4, filtered, and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.72) to give compound SM12 as a yellow solid (0.11 g, 36%):1 1H NMR (500 MHz, CDCl3) δ 8.45 (d, J = 2.2 Hz, 1H, aromatic), 8.08 (dd, J1 = 9.1 Hz, J2 = 2.2 Hz, 1H, aromatic), 7.68 (d, J = 2.5 Hz, 1H, aromatic), 7.34 (dd, J1 = 8.6 Hz, J2 = 2.5 Hz, 1H, aromatic), 7.28 (d, J = 9.1 Hz, 1H, aromatic), 6.90 (d, J = 8.6 Hz, 1H, aromatic), 6.87 (app.t, J = 1.1 Hz, 1H, aromatic), 4.06 (s, 2H, C H 2Ar), 3.87 (d, J = 7.4 Hz, 2H, NC H 2), 3.87 (s, 3H, ArOC H 3), 3.84 (s, 3H, ArCO2C H 3), 2.14 (septet, J = 7.0 Hz, 1H, C H (CH3)2), 0.90 (t, J = 6.7 Hz, 6H, CH(C H 3)2); 13 13C NMR (100 MHz, CDCl3) δ 166.9, 157.9, 141.3, 139.9, 133.7, 132.1, 131.9, 129.9, 127.1, 120.2, 117.6, 117.4, 116.8, 112.5, 109.7, 56.3, 54.5, 52.2, 30.3, 29.9, 20.4.

[0202] Synthesis of compound SM13. A solution of compound SM6 (0.70 g, 2.77 mmol) and methyl 5-formyl-2-methoxybenzoate (1.08 g, 2.77 mmol) in anhydrous CH2Cl2 (5 mL) was cooled to 0 °C in an ice-water bath. Then Et3SiH (1.2 mL, 7.77 mmol) was added, followed by TFA (0.4 mL, 5.55 mmol). The mixture was stirred at 0 °C for 10 minutes and allowed to warm to room temperature overnight. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with NaHCO3, H2O, and brine, dried over MgSO4, filtered, and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.59) to give compound SM13 (0.23 g, 20%) as a yellow solid: 11H NMR (500 MHz, CDCl3) δ 8.47 (dd, J1 = 2.3 Hz, J2 = 0.5 Hz, 1H, aromatic), 8.05 (dd, J1 = 9.1 Hz, J2 = 2.2 Hz, 1H, aromatic), 7.68 (d, J = 2.4 Hz, 1H, aromatic), 7.35 (dd, J1 = 8.6 Hz, J2 = 2.5 Hz, 1H, aromatic), 7.32 - 7.27 (m, 3H, aromatic), 7.08 - 7.05 (m, 2H, aromatic), 6.95 (app.t, J = 1.1 Hz, 1H, aromatic), 6.90 (d, J = 8.6 Hz, 1H, aromatic), 5.28 (s, 2H, NC H 2Ar), 4.07 (s, 2H, C H 2Ar), 3.87 (s, 3H, ArOC H 3), 3.85 (s, 3H, ArCO2C H 3); 13 13C NMR (100 MHz, CDCl3) δ 166.9, 157.9, 141.6, 139.8, 136.5, 133.7, 131.93, 131.87, 129.8, 129.2, 128.3, 127.5, 126.9, 120.3, 118.0, 117.9, 116.8, 112.5, 109.9, 56.3, 52.2, 50.7, 30.4.

[0203]

[0204] Synthesis of compound SM14. A solution of methyl 2 - methoxy - 5 - ((1 - methyl - 5 - nitro - 1H - indol - 3 - yl)methyl)benzoate (112 mg, 0.32 mmol) in MeOH / THF / H2O (3 mL / 1 mL / 0.6 mL) was treated with KOH pellets (124 mg, 2.21 mmol) and the mixture was refluxed at 65 °C for 2 h. After completion of the reaction, the organic solvents were removed in vacuo. The resulting mixture was acidified to pH 1 with 1 N aqueous HCl. The precipitate was filtered and eluted with H2O to give compound 20 as a yellow solid (94 mg, 87%): 11H NMR (400 MHz, CDCl3) δ 10.7 (very broad s, 1H, CO2H), 8.38 (d, J = 2.0 Hz, 1H, aromatic), 8.10 (dd, J1 = 8.8 Hz, J2 = 2.0 Hz, 1H, aromatic), 8.04 (d, J = 2.4 Hz, 1H, aromatic), 7.49 (dd, J1 = 8.4 Hz, J2 = 2.0 Hz, 1H, aromatic), 7.28 (d, J = 9.2 Hz, 1H, aromatic), 7.00 (d, J = 8.0 Hz, 1H, aromatic), 6.94 (s, 1H, aromatic), 4.09 (s, 2H, CH2Ar), 4.05 (s, 3H, ArOCH3), 3.79 (s, 3H, NCH3); 13 13C NMR (100 MHz, CDCl3) δ 165.2, 156.5, 141.2, 140.0, 135.0, 134.4, 133.6, 130.3, 126.8, 117.6, 117.5, 116.5, 116.4, 111.9, 109.2, 56.8, 33.2, 30.1; C 18 H 16 Calculated m / z for N2O5 340.1; found 323.1 [M - OH] + 。

[0205] Synthesis of compound SM15. A solution of compound SM9 (0.23 g, 0.62 mmol) in MeOH:THF:H2O / 10:2:2 (total 14 mL) was stirred and treated with KOH pellets (0.25 g, 4.37 mmol). The resulting mixture was refluxed at 65 °C for 2 h. After completion of the reaction, the organic solvents were removed in vacuo. The resulting mixture was acidified to pH 1 with 1N aqueous HCl and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated to give compound SM15 as a yellow solid (0.17 g, 75%): 1 1H NMR (500 MHz, CDCl3) δ 8.38 (d, J = 2.2 Hz, 1H, aromatic), 8.09 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 8.06 (d, J = 2.5 Hz, 1H, aromatic), 7.49 (dd, J1 = 8.5 Hz, J2 = 2.5 Hz, 1H, aromatic), 7.30 (d, J = 9.2 Hz, 1H, aromatic), 7.00 (d, J = 8.2 Hz, 1H, aromatic), 6.99 (s, 1H, aromatic), 4.15 (q, J = 7.4 Hz, 2H, NC H 2CH3), 4.10 (s, 2H, CH 2Ar), 4.05 (s, 3H, ArOC H 3), 1.46 (t, J = 7.3 Hz, 3H, NCH2C H 3); 13 C NMR (100 MHz, CDCl3) δ 165.5, 156.8, 135.3, 134.7, 133.9, 128.7, 127.1, 117.8, 117.6, 116.9, 116.7, 112.1, 109.4, 57.0, 41.6, 30.5, 15.6.

[0206] Synthesis of compound SM16. Stir a solution of compound SM10 (0.20 g, 0.52 mmol) in MeOH:THF:H2O / 10:2:2 (total 14 mL) and treat with KOH pellets (0.21 g, 3.66 mmol). The resulting mixture was refluxed at 65 °C for 2 h. After completion of the reaction, the organic solvents were removed in vacuo. The resulting mixture was acidified to pH 1 with 1N aqueous HCl and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated to give compound SM16 as a yellow solid (0.17 g, 88%): 1 H NMR (500 MHz, CDCl3) δ 8.38 (d, J = 2.2 Hz, 1H, aromatic), 8.08 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 8.06 (d, J = 2.5 Hz, 1H, aromatic), 7.48 (dd, J1 = 8.5 Hz, J2 = 2.5 Hz, 1H, aromatic), 7.30 (d, J = 9.2 Hz, 1H, aromatic), 6.99 (d, J = 8.6 Hz, 1H, aromatic), 6.97 (app.t, J = 1.0 Hz, 1H, aromatic), 4.10 (s, 2H, C H 2Ar), 4.06 (t, J = 7.2 Hz, 2H, NC H 2CH2CH3), 4.05 (s, 3H, ArOC H 3), 1.85 (sextet, J = 7.4 Hz, 2H, NCH2C H 2CH3), 0.92 (t, J = 7.4 Hz, 3H, NCH2CH2C H 3); 1313C NMR (100 MHz, CDCl3) δ 165.5, 156.8, 141.3, 139.7, 135.2, 134.6, 133.8, 129.5, 127.0, 117.8, 117.6, 116.68, 116.66, 112.1, 109.6, 57.0, 48.6, 30.4, 23.8, 11.7。

[0207] Synthesis of compound SM17. Stir a solution of compound SM11 (90 mg, 0.23 mmol) in MeOH:THF:H2O / 5:1:1 (total 7 mL) and treat with KOH pellets (89 mg, 1.59 mmol). Reflux the resulting mixture at 65 °C for 2 h. After completion of the reaction, remove the organic solvents in vacuo. Acidify the resulting mixture to pH 1 with 1 N aqueous HCl and extract with CH2Cl2 (3×). Wash the combined organic layers with brine, dry over MgSO4, filter, and concentrate to give compound SM17 (71 mg, 82%) as a yellow solid: 1 1H NMR (500 MHz, CD3OD) δ 8.30 (dd, J1 = 2.3 Hz, J2 = 0.6 Hz, 1H, aromatic), 7.97 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.66 (d, J = 2.4 Hz, 1H, aromatic), 7.42 (d, J = 9.1 Hz, 1H, aromatic), 7.39 (dd, J1 = 8.6 Hz, J2 = 2.5 Hz, 1H, aromatic), 7.17 (app.t, J = 1.0 Hz, 1H, aromatic), 6.99 (d, J = 8.6 Hz, 1H, aromatic), 4.13 (t, J = 7.1 Hz, 2H, NC H 2CH2CH2CH3), 4.04 (s, 2H, C H 2Ar), 3.91 (s, 1H, O H ), 3.81 (s, 3H, ArOC H 3), 1.76 - 1.70 (m, 2H, NCH2C H 2CH2CH3), 1.27 - 1.20 (m, 2H, NCH2CH2C H 2CH3), 0.86 (t, J = 7.5 Hz, 3H, NCH2CH2CH2C H 3); 1313C NMR (100 MHz, (CD3)2SO) δ 156.4, 140.2, 139.1, 132.8, 132.6, 132.5, 130.4, 126.4, 121.0, 116.6, 116.5, 116.0, 113.0, 112.5, 110.4, 55.8, 45.5, 31.9, 29.1, 19.4, 13.5.

[0208] Synthesis of compound SM18. A solution of compound SM12 (0.10 g, 0.25 mmol) in MeOH:THF:H2O / 10:2:2 (total 14 mL) was stirred and treated with KOH pellets (99 mg, 1.77 mmol). The resulting mixture was refluxed at 65 °C for 2 h. After completion of the reaction, the organic solvents were removed in vacuo. The resulting mixture was acidified to pH 1 with 1 N aqueous HCl and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated to give compound SM18 (81 mg, 84%) as a yellow solid (containing some methyl 5-formyl-2-methoxybenzoate, which was removed in the next synthetic step): 1 1H NMR (500 MHz, (CD3)2SO) δ 8.41 (d, J = 2.3 Hz, 1H, aromatic), 8.00 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.68 (d, J = 9.2 Hz, 1H, aromatic), 7.54 (d, J = 2.5 Hz, 1H, aromatic), 7.47 (s, 1H, aromatic), 7.42 (dd, J1 = 8.6 Hz, J2 = 2.4 Hz, 1H, aromatic), 7.05 (d, J = 8.6 Hz, 1H, aromatic), 4.10 (s, 2H, C H 2Ar), 4.04 (d, J = 7.4 Hz, 2H, NC H 2), 3.77 (s, 3H, ArOC H 3), 2.10 (septet, J = 7.2 Hz, 1H, C H (CH3)2), 0.84 (d, J = 6.7 Hz, 6H, CH(C H 3)2); 13 13C NMR (100 MHz, (CD3)2SO) δ 166.4, 162.6, 140.2, 134.3, 132.8, 132.54, 132.45, 130.9, 128.7, 121.8, 121.0, 116.5, 116.0, 113.0, 112.5, 110.7, 56.4, 55.8, 29.3, 19.7 (2CH3).

[0209] Synthesis of compound SM19. A solution of compound SM13 (0.20 g, 0.46 mmol) in MeOH:THF:H2O / 10:2:2 (total 14 mL) was stirred and treated with KOH pellets (0.18 g, 3.25 mmol). The resulting mixture was refluxed at 65 °C for 2 h. After completion of the reaction, the organic solvents were removed in vacuo. The resulting mixture was acidified to pH 1 with 1 N aqueous HCl and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated to give compound SM19 as a yellow solid (60 mg, 31%): 1 H NMR (500 MHz, CD3OD) δ 8.33 (d, J = 2.1 Hz, 1H, aromatic), 7.94 (dd, J1 = 9.2 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.64 (d, J = 2.5 Hz, 1H, aromatic), 7.39 (dd, J1 = 8.5 Hz, J2 = 2.6 Hz, 1H, aromatic), 7.37 (d, J = 9.1 Hz, 1H, aromatic), 7.24 - 7.20 (m, 3H, aromatic), 7.20 - 7.15 (m, 1H, aromatic), 7.09 - 7.07 (m, 2H, aromatic), 6.99 (d, J = 8.6 Hz, 1H, aromatic), 5.35 (s, 2H, NC H 2Ar), 4.06 (s, 2H, C H 2Ar), 3.90 (s, 1H, O H ), 3.80 (s, 3H, ArOC H 3); 13 C NMR (100 MHz, CDCl3) δ 165.5, 156.8, 141.6, 139.9, 136.4, 135.2, 134.5, 133.8, 130.0, 129.3, 128.3, 127.3, 126.9, 118.0, 117.8, 117.2, 116.7, 112.1, 110.0, 57.0, 50.7, 30.4。

[0210] Synthesis of Compound 27. A solution of compound SM14 (2-methoxy-5-((1-methyl-5-nitro-1H-indol-3-yl)methyl)benzoic acid) (30 mg, 0.09 mmol), 2-chlorobenzenesulfonamide (20b; 20 mg, 0.10 mmol), EDC·HCl (27 mg, 0.14 mmol) and DMAP (19 mg, 0.16 mmol) in anhydrous CH2Cl2 (3 mL) was stirred overnight at room temperature. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.17) to give compound 27 (18 mg, 40%) as an orange solid: 1 1H NMR (500 MHz, CDCl3) δ 10.66 (s, 1H, N H ), 8.29 (dd, J1 = 7.9 Hz, J2 = 1.7 Hz, 1H, aromatic), 8.20 (d, J = 2.2 Hz, 1H, aromatic), 8.09 (dd, J1 = 9.1 Hz, J2 = 2.2 Hz, 1H, aromatic), 7.89 (d, J = 2.5 Hz, 1H, aromatic), 7.56 (dd, J1 = 8.6 Hz, J2 = 2.5 Hz, 1H, aromatic), 7.53 - 7.42 (m, 3H, aromatic), 7.29 (d, J = 9.1 Hz, 1H, aromatic), 7.03 (d, J = 8.6 Hz, 1H, aromatic), 6.55 (s, 1H, aromatic), 4.08 (s, 3H, NC H 3), 3.71 (s, 5H, C H 2Ar, ArOC H 3); 13 13C NMR (100 MHz, (CD3)2SO) δ 155.5, 140.3, 139.8, 135.9, 135.2, 133.1, 132.3, 131.8 (2CH), 131.7, 130.8, 128.9, 127.61, 127.59, 125.8 (2CH), 119.8, 116.7, 116.3, 112.3, 110.5, 56.0, 36.6, 32.9; C 24 H 20 Calculated m / z for C19H16ClN3O6S: 513.95.

[0211] Synthesis of Compound 28 (SGT1650). A solution of Compound SM14KCH-2-127 (30 mg, 0.09 mmol), 2-bromobenzenesulfonamide (20c; 25 mg, 0.10 mmol), EDC·HCl (27 mg, 0.14 mmol), and DMAP (19 mg, 0.16 mmol) in anhydrous CH2Cl2 (3 mL) was stirred overnight at room temperature. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.25) to give Compound 28 (26 mg, 53%) as an orange solid: 1 H NMR (500 MHz, CDCl3) δ 10.72 (s, 1H, N H ), 8.33 (dd, J1 = 7.9 Hz, J2 = 1.7 Hz, 1H, aromatic), 8.20 (d, J = 2.2 Hz, 1H, aromatic), 8.10 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.89 (d, J = 2.5 Hz, 1H, aromatic), 7.69 (dd, J1 = 7.9 Hz, J2 = 1.3 Hz, 1H, aromatic), 7.56 (dd, J1 = 8.6 Hz, J2 = 2.6 Hz, 1H, aromatic), 7.49 (td, J1 = 7.7 Hz, J2 = 1.3 Hz, 1H, aromatic), 7.42 (qd, J1 = 8.5 Hz, J2 = 1.8 Hz, 1H, aromatic), 7.29 (d, J = 9.1 Hz, 1H, aromatic), 7.03 (d, J = 8.6 Hz, 1H, aromatic), 6.55 (s, 1H, aromatic), 4.08 (s, 3H, NC H 3), 3.71 (s, 5H, C H 2Ar, ArOC H 3); 13 C NMR (100 MHz, (CD3)2SO) δ 155.6, 140.3, 139.8, 136.0, 135.2, 132.6, 131.9 (2CH), 129.1, 128.1, 125.8 (2CH), 119.8, 119.2 (2C), 116.7, 116.3, 112.4, 110.5 (2CH), 109.6, 56.1, 36.6, 32.9; C 24 H 20 Calculated m / z for C18H14BrN3O6S: 558.40.

[0212] Synthesis of Compound 29. A solution of compound SM14 (30 mg, 0.09 mmol), 2,6-difluorobenzenesulfonamide (20d; 20 mg, 0.10 mmol), EDC·HCl (27 mg, 0.14 mmol), and DMAP (19 mg, 0.16 mmol) in anhydrous CH2Cl2 (3 mL) was stirred overnight at room temperature. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.29) to give Compound 29 (11 mg, 25%) as a yellow solid: 1 1H NMR (500 MHz, CDCl3) δ 10.64 (s, 1H, N H ), 8.33 (d, J = 2.8 Hz, 1H, aromatic), 8.08 (dd, J1 = 9.3 Hz, J2 = 2.4 Hz, 1H, aromatic), 7.90 (d, J = 2.4 Hz, 1H, aromatic), 7.56 - 7.51 (m, 1H, aromatic), 7.48 (dd, J1 = 8.6 Hz, J2 = 2.6 Hz, 1H, aromatic), 7.26 (d, J = 9.2 Hz, 1H, aromatic), 7.01 (t, J = 8.7 Hz, 2H, aromatic), 6.84 (d, J = 8.6 Hz, 1H, aromatic), 6.88 (s, 1H, aromatic), 4.05 (s, 3H, NC H 3), 4.03 (s, 2H, C H 2Ar), 3.76 (s, 3H, ArOC H 3); 13 13C NMR (100 MHz, (CD3)2SO) δ 164.1, 157.9, 148.8 (2C), 140.2, 139.6, 131.4, 129.2, 129.13 (2CH), 129.10, 129.06, 126.3, 126.0, 117.5, 116.6, 116.5, 115.9 (2CH), 112.2, 110.4, 55.9, 43.5, 32.8; C 24 1H 19 Calculated m / z for C18H15F2N3O6S: 515.49.

[0213] Synthesis of Compound 30. A solution of Compound SM14 (30 mg, 0.09 mmol), 2,4-difluorobenzenesulfonamide (20e; 20 mg, 0.10 mmol), EDC·HCl (27 mg, 0.14 mmol) and DMAP (19 mg, 0.16 mmol) in anhydrous CH2Cl2 (3 mL) was stirred overnight at room temperature. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.41) to give Compound 30 as a yellow solid (16 mg, 35%): 1 1H NMR (500 MHz, CDCl3) δ 10.57 (s, 1H, N H ), 8.33 (d, J = 2.2 Hz, 1H, aromatic), 8.18 - 8.14 (m, 1H, aromatic), 8.08 (dd, J1 = 9.1 Hz, J2 = 2.4 Hz, 1H, aromatic), 7.87 (d, J = 2.5 Hz, 1H, aromatic), 7.47 (dd, J1 = 8.5 Hz, J2 = 2.1 Hz, 1H, aromatic), 7.26 (d, J = 9.1 Hz, 1H, aromatic), 7.05 - 7.01 (m, 1H, aromatic), 6.97 (d, J = 8.6 Hz, 1H, aromatic), 6.92 - 6.88 (m, 1H, aromatic), 6.87 (s, 1H, aromatic), 4.05 (s, 3H, NC H 3), 4.03 (s, 2H, C H 2Ar), 3.76 (s, 3H, ArOC H 3); 13 13C NMR (100 MHz, (CD3)2SO) δ 155.3, 140.2, 139.6 (2C), 133.6, 133.5, 132.9, 131.4, 129.1, 126.3, 116.5, 116.4, 115.9, 112.3, 112.2, 110.4 (2CH), 106.2, 105.94, 105.91, 105.7, 55.9, 32.8, 28.9; C 24 H 19 Calculated m / z for C18H15F2N3O6S: 515.49.

[0214] Synthesis of Compound 31. A solution of Compound SM14 (30 mg, 0.09 mmol), 3,4-difluorobenzenesulfonamide (20f; 20 mg, 0.10 mmol), EDC·HCl (27 mg, 0.14 mmol) and DMAP (19 mg, 0.16 mmol) in anhydrous CH2Cl2 (3 mL) was stirred overnight at room temperature. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.18) to give Compound 31 (12 mg, 26%) as an amber solid: 1 H NMR (500 MHz, CDCl3) δ 10.43 (s, 1H, N H ), 8.19 (d, J = 2.0 Hz, 1H, aromatic), 8.10 (dd, J1 = 9.1 Hz, J2 = 2.2 Hz, 1H, aromatic), 7.99 - 7.94 (m, 2H, aromatic), 7.92 - 7.89 (m, 1H, aromatic), 7.54 (dd, J1 = 8.6 Hz, J2 = 2.5 Hz, 1H, aromatic), 7.31 (d, J = 9.1 Hz, 2H, aromatic), 7.00 (d, J = 8.7 Hz, 1H, aromatic), 6.58 (s, 1H, aromatic), 4.06 (s, 3H, NC H 3), 3.73 (s, 5H, C H 2Ar, ArOC H 3); 13 C NMR (100 MHz, (CD3)2SO) δ 172.0, 164.1, 154.9 (2C), 140.2, 139.8, 134.6, 131.8, 131.7, 131.5, 128.3, 128.2, 125.9, 120.4 (2CH), 117.5, 116.8, 116.62, 116.58, 116.4, 110.5, 55.5, 37.1, 32.9; C 24 H 19 Calculated m / z for C19H16F2N3O6S: 515.49.

[0215] Synthesis of Compound 32. A solution of Compound SM15 (30 mg, 0.08 mmol), 2-fluorobenzenesulfonamide (20a; 18 mg, 0.10 mmol), EDC·HCl (26 mg, 0.14 mmol) and DMAP (19 mg, 0.15 mmol) in anhydrous CH2Cl2 (3 mL) was stirred overnight at room temperature. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.46) to give Compound 32 (50 mg, quantitative) as a yellow solid: 1 1H NMR (500 MHz, (CD3)2SO) δ 12.25 (s, 1H, N H ), 8.43 (dd, J1 = 2.4 Hz, J2 = 0.5 Hz, 1H, aromatic), 8.00 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.95 (t, J = 7.7 Hz, 1H, aromatic), 7.81 - 7.75 (m, 1H, aromatic), 7.65 (dd, J1 = 9.2 Hz, J2 = 0.5 Hz, 1H, aromatic), 7.49 (s, 1H, aromatic), 7.47 - 7.43 (m, 1H, aromatic), 7.30 (d, J = 2.3 Hz, 1H, aromatic), 7.06 (d, J = 8.6 Hz, 1H, aromatic), 4.24 (q, J = 7.3 Hz, 2H, NC H 2CH3), 4.07 (s, 2H, C H 2Ar), 3.78 (s, 3H, ArOC H 3), 1.35 (t, J = 7.3 Hz, 3H, NCH2C H 3); 13 13C NMR (100 MHz, (CD3)2SO) δ 164.0, 157.0, 155.3, 140.2, 138.7, 132.9, 131.2, 129.8, 129.1, 126.4, 124.7, 117.5, 117.2, 117.0, 116.6, 116.4, 116.0, 112.2, 110.3, 109.6, 55.9, 40.7, 29.0, 15.4; C 25 H 22 Calculated m / z for C18H15FN3O6S: 511.52.

[0216] Synthesis of Compound 38. A solution of Compound SM15 (30 mg, 0.08 mmol), 4-nitrobenzenesulfonamide (20 g; 20 mg, 0.10 mmol), EDC·HCl (26 mg, 0.14 mmol), and DMAP (19 mg, 0.15 mmol) in anhydrous CH2Cl2 (3 mL) was stirred overnight at room temperature. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.27) to give Compound 38 (37 mg, 82%) as an orange solid: 1 H NMR (500 MHz, (CD3)2SO) δ 12.19 (s, 1H, N H ), 8.45 (d, J = 8.9 Hz, 2H, aromatic), 8.40 (d, J = 2.3 Hz, 1H, aromatic), 8.20 (d, J = 9.0 Hz, 2H, aromatic), 7.99 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.64 (d, J = 9.1 Hz, 1H, aromatic), 7.48 (s, 1H, aromatic), 7.46 (dd, J1 = 8.6 Hz, J2 = 2.4 Hz, 1H, aromatic), 7.33 (d, J = 2.3 Hz, 1H, aromatic), 7.06 (d, J = 8.6 Hz, 1H, aromatic), 4.23 (q, J = 7.3 Hz, 2H, NC H 2CH3), 4.06 (s, 2H, C H 2Ar), 3.80 (s, 3H, ArOC H 3), 1.33 (t, J = 7.3 Hz, 3H, NCH2C H 3); 13 C NMR (100 MHz, (CD3)2SO) δ 165.1, 155.5, 150.3, 144.6, 140.2, 138.6, 133.5, 133.0, 129.8, 129.4, 129.3, 126.4, 124.4, 121.6, 116.5, 116.4, 115.9, 112.3, 110.3, 56.0, 40.7, 28.9, 15.3; C 25 H 22 Calculated m / z for C

[0217] Synthesis of Compound 33. A solution of Compound SM16 (30 mg, 0.08 mmol), 2-fluorobenzenesulfonamide (20a; 17 mg, 0.10 mmol), EDC·HCl (25 mg, 0.13 mmol) and DMAP (18 mg, 0.15 mmol) in anhydrous CH2Cl2 (3 mL) was stirred overnight at room temperature. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f = 0.46) to give Compound 33 (39 mg, 91%) as a yellow solid: 1 1H NMR (500 MHz, (CD3)2SO) δ 12.25 (s, 1H, N H ), 8.42 (d, J = 2.3 Hz, 1H, aromatic), 7.99 (dd, J1 = 9.2 Hz, J2 = 2.4 Hz, 1H, aromatic), 7.95 (t, J = 7.3 Hz, 1H, aromatic), 7.81 - 7.74 (m, 1H, aromatic), 7.66 (d, J = 9.2 Hz, 1H, aromatic), 7.47 (s, 1H, aromatic), 7.44 (m, 3H, aromatic), 7.29 (d, J = 2.3 Hz, 1H, aromatic), 7.05 (d, J = 8.6 Hz, 1H, aromatic), 4.17 (t, J = 7.0 Hz, 2H, NC H 2CH2CH3), 4.07 (s, 2H, C H 2Ar), 3.78 (s, 3H, ArOC H 3), 1.79 (sextet, J = 7.3 Hz, 2H, NCH2C H 2CH3), 0.80 (t, J = 7.4 Hz, 3H, NCH2CH2C H 3); 13 13C NMR (100 MHz, (CD3)2SO) δ 159.5, 157.0, 155.3, 140.2, 139.1, 136.5, 133.1, 132.9, 131.2, 130.4, 129.1, 126.3, 124.8, 124.7, 117.2, 117.0, 116.44, 116.37, 115.9, 112.2, 110.4, 59.7, 55.9, 47.3, 28.9, 23.1, 11.0; C 26 H 24 Calculated m / z for C21H20FN3O6S: 525.55.

[0218] Synthesis of Compound 39. A solution of Compound SM16 (30 mg, 0.08 mmol), 4-nitrobenzenesulfonamide (20 g; 19 mg, 0.10 mmol), EDC·HCl (25 mg, 0.13 mmol) and DMAP (18 mg, 0.15 mmol) in anhydrous CH2Cl2 (3 mL) was stirred overnight at room temperature. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.26) to give Compound 39 (22 mg, 49%) as an orange solid: 1 H NMR (500 MHz, CDCl3) δ 10.49 (s, 1H, N H ), 8.35 (d, J = 9.1 Hz, 2H, aromatic), 8.32 (d, J = 8.8 Hz, 2H, aromatic), 8.30 (d, J = 2.3 Hz, 1H, aromatic), 8.06 (dd, J1 = 9.1 Hz, J2 = 2.2 Hz, 1H, aromatic), 7.90 (d, J = 2.4 Hz, 1H, aromatic), 7.45 (dd, J1 = 8.6 Hz, J2 = 2.5 Hz, 1H, aromatic), 7.28 (d, J = 9.1 Hz, 1H, aromatic), 6.95 (d, J = 8.6 Hz, 1H, aromatic), 6.93 (s, 1H, aromatic), 4.03 (t, J = 7.2 Hz, 2H, NC H 2CH2CH3), 4.04 (s, 2H, C H 2Ar), 4.02 (s, 3H, ArOC H 3), 1.83 (sextet, J = 7.3 Hz, 2H, NCH2C H 2CH3), 0.90 (t, J = 7.4 Hz, 3H, NCH2CH2C H 3); 13 C NMR (100 MHz, CDCl3) δ 162.6, 156.7, 150.9, 144.6, 141.3, 139.6, 135.8, 134.4, 132.7, 130.3, 129.5, 127.0, 124.2, 118.3, 117.6, 116.6, 116.5, 112.3, 109.6, 56.9, 48.6, 30.3, 23.8, 11.7; C 26 H 24 Calculated m / z for C

[0219] Synthesis of Compound 34. A solution of compound SM17 (30 mg, 0.08 mmol), 2-fluorobenzenesulfonamide (20a; 16 mg, 0.09 mmol), EDC·HCl (24 mg, 0.13 mmol), and DMAP (17 mg, 0.14 mmol) in anhydrous CH2Cl2 (3 mL) was stirred overnight at room temperature. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.63) to give compound 34 (24 mg, 56%) as a yellow solid: 1 1H NMR (500 MHz, CDCl3) δ 10.57 (s, 1H, N H ), 8.33 (d, J = 2.3 Hz, 1H, aromatic), 8.14 (td, J1 = 7.6 Hz, J2 = 1.8 Hz, 1H, aromatic), 8.06 (dd, J1 = 9.1 Hz, J2 = 2.2 Hz, 1H, aromatic), 7.88 (d, J = 2.5 Hz, 1H, aromatic), 7.61 - 7.57 (m, 1H, aromatic), 7.44 (dd, J1 = 8.6 Hz, J2 = 2.6 Hz, 1H, aromatic), 7.33 - 7.30 (m, 1H, aromatic), 7.27 (d, J = 9.1 Hz, 1H, aromatic), 7.17 - 7.13 (m, 1H, aromatic), 6.96 (d, J = 8.6 Hz, 1H, aromatic), 6.89 (s, 1H, aromatic), 4.05 (t, J = 7.3 Hz, 2H, NC H 2CH2CH2CH3), 4.05 (s, 3H, ArOC H 3), 4.02 (s, 2H, C H 2Ar), 1.76 (p, J = 7.6 Hz, 2H, NCH2C H 2CH2CH3), 1.30 (sextet, J = 7.7 Hz, 2H, NCH2CH2C H 2CH3), 0.91 (t, J = 7.4 Hz, 3H, NCH2CH2CH2C H 3); 1313C NMR (100 MHz, CDCl3) δ 162.7, 156.7, 141.3, 139.6, 136.4, 136.3, 135.5, 134.2, 132.6, 132.3, 129.4, 127.0, 124.7, 124.6, 118.8, 117.6, 117.3, 117.1, 116.6, 112.3, 109.5, 56.9, 46.7, 32.5, 30.3, 20.3, 13.8; C 27 H 26 Calculated m / z for FN3O6S: 539.58.

[0220] Synthesis of Compound 40. A solution of Compound SM17 (30 mg, 0.08 mmol), 4-nitrobenzenesulfonamide (20 g; 19 mg, 0.09 mmol), EDC·HCl (24 mg, 0.13 mmol), and DMAP (17 mg, 0.14 mmol) in anhydrous CH2Cl2 (3 mL) was stirred overnight at room temperature. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.45) to give Compound 40 (28 mg, 62%) as a yellow solid: 1 1H NMR (500 MHz, CDCl3) δ 10.49 (s, 1H, N H ), 8.35 (d, J = 9.0 Hz, 2H, aromatic), 8.32 (d, J = 9.1 Hz, 2H, aromatic), 8.29 (d, J = 2.3 Hz, 1H, aromatic), 8.05 (dd, J1 = 9.1 Hz, J2 = 2.2 Hz, 1H, aromatic), 7.90 (d, J = 2.5 Hz, 1H, aromatic), 7.45 (dd, J1 = 8.6 Hz, J2 = 2.5 Hz, 1H, aromatic), 7.28 (d, J = 9.1 Hz, 1H, aromatic), 6.95 (d, J = 8.6 Hz, 1H, aromatic), 6.92 (s, 1H, aromatic), 4.06 (t, J = 7.2 Hz, 1H, aromatic), 4.04 (s, 2H, C H 2Ar), 4.03 (s, 3H, ArOC H 3), 1.77 (p, J = 7.5 Hz, 2H, NCH2C H 2CH2CH3), 1.30 (sextet, J = 7.6 Hz, 2H, NCH2CH2C H2CH3), 0.92 (t, J=7.4 Hz, 3H, NCH2CH2CH2C H 3); 13 C NMR (100 MHz, CDCl3) δ 162.6, 156.7, 150.9, 144.6, 141.3, 139.6, 135.8, 134.4, 132.6, 130.3, 129.4, 126.9, 124.2, 118.3, 117.6, 116.6, 116.5, 112.3, 109.6, 56.9, 46.7, 32.5, 30.3, 20.3, 13.8; C 27 H 26 Calculated m / z for C14H10N4O8S: 566.59.

[0221] Synthesis of compound 35. A solution of compound SM18 (30 mg, 0.08 mmol), 2-fluorobenzenesulfonamide (20a; 16 mg, 0.09 mmol), EDC·HCl (24 mg, 0.13 mmol) and DMAP (17 mg, 0.14 mmol) in anhydrous CH2Cl2 (3 mL) was stirred overnight at room temperature. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.29) to give compound 35 (22 mg, 52%) as a yellow solid: 1 1H NMR (500 MHz, (CD3)2SO) δ 12.24 (s, 1H, N H ), 8.41 (d, J=7.7 Hz, 1H, aromatic), 7.98 (dd, J1=9.1 Hz, J2=2.3 Hz, 1H, aromatic), 7.95 (t, J=7.7 Hz, 1H, aromatic), 7.82 - 7.75 (m, 1H, aromatic), 7.67 (d, J=9.1 Hz, 1H, aromatic), 7.45 (s, 1H, aromatic), 7.44 (m, 3H, aromatic), 7.28 (d, J=2.3 Hz, 1H, aromatic), 7.06 (d, J=8.6 Hz, 1H, aromatic), 4.07 (s, 2H, C H 2Ar), 4.03 (d, J=7.3 Hz, 2H, NC H 2), 3.78 (s, 3H, ArOC H 3), 2.09 (septet, J=7.2 Hz, 1H, C H (CH3)2), 0.82 (d, J=6.7 Hz, 6H, CH(CH 3)2); 13 C NMR (100 MHz, (CD3)2SO) δ 164.2, 159.6, 155.3, 140.2, 139.4, 136.6, 133.0, 131.2, 130.9, 129.1, 126.3, 124.8, 117.4, 117.3, 117.1, 116.5, 116.3, 115.9, 115.3, 112.2, 110.7, 55.9, 52.9, 29.2, 28.9, 19.7; C 27 H 26 Calculated m / z for FN3O6S: 539.58.

[0222] Synthesis of Compound 41. A solution of Compound SM18 (30 mg, 0.08 mmol), 4-nitrobenzenesulfonamide (20 g; 19 mg, 0.09 mmol), EDC·HCl (24 mg, 0.13 mmol), and DMAP (17 mg, 0.14 mmol) in anhydrous CH2Cl2 (3 mL) was stirred overnight at room temperature. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.41) to give Compound 41 (5 mg, 12%) as a yellow solid: 1 H NMR (500 MHz, CD3OD) δ 8.40 (d, J = 8.9 Hz, 2H, aromatic), 8.31 (d, J = 2.2 Hz, 1H, aromatic), 8.27 (d, J = 9.0 Hz, 2H, aromatic), 8.02 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.49 (d, J = 9.2 Hz, 1H, aromatic), 7.48 (d, J = 2.2 Hz, 1H, aromatic), 7.45 (dd, J1 = 8.7 Hz, J2 = 1.8 Hz, 1H, aromatic), 7.23 (s, 1H, aromatic), 7.06 (d, J = 8.5 Hz, 1H, aromatic), 4.09 (s, 2H, C H 2Ar), 3.99 (d, J = 7.4 Hz, 2H, NC H 2), 3.91 (s, 3H, ArOC H 3), 2.15 (septet, J = 7.3 Hz, 1H, C H (CH3)2), 0.89 (d, J = 6.7 Hz, 6H, CH(C H (CH3)2); 1313C NMR (100 MHz, (CD3)2SO) δ 165.1, 155.5, 150.2, 144.6, 140.1, 139.4, 133.4, 133.0, 130.9, 129.32, 129.26, 126.2, 124.4, 121.6, 116.4, 116.2, 115.9, 112.2, 110.6, 56.0, 52.9, 29.2, 28.8, 19.7; C 27 H 26 Calculated m / z for C14H12N4O8S: 566.59.

[0223] Synthesis of Compound 42. A solution of Compound SM19 (30 mg, 0.07 mmol), 4-nitrobenzenesulfonamide (20 mg; 17 mg, 0.09 mmol), EDC·HCl (22 mg, 0.12 mmol), and DMAP (16 mg, 0.13 mmol) in anhydrous CH2Cl2 (3 mL) was stirred overnight at room temperature. The reaction was quenched with H2O and extracted with CH2Cl2 (3×). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated. The resulting crude product was purified by column chromatography (silica gel, hexane:EtOAc / 1:1, R f 0.42) to afford Compound 42 (29 mg, 66%) as a yellow solid: 1 1H NMR (500 MHz, CDCl3) δ 10.49 (s, 1H, N H ), 8.35 (d, J = 9.1 Hz, 2H, aromatic), 8.32 (d, J = 9.1 Hz, 2H, aromatic), 8.31 (d, J = 2.3 Hz, 1H, aromatic), 8.02 (dd, J1 = 9.1 Hz, J2 = 2.3 Hz, 1H, aromatic), 7.90 (d, J = 2.4 Hz, 1H, aromatic), 7.46 (dd, J1 = 8.6 Hz, J2 = 2.5 Hz, 1H, aromatic), 7.32 - 7.27 (m, 3H, aromatic), 7.23 (d, J = 9.5 Hz, 1H, aromatic), 7.05 (dd, J1 = 7.9 Hz, J2 = 2.2 Hz, 2H, aromatic), 6.97 (s, 1H, aromatic), 6.96 (d, J = 8.7 Hz, 1H, aromatic), 5.27 (s, 2H, NC H 2Ar), 4.05 (s, 2H, C H 2Ar), 4.04 (s, 3H, ArOC H 3); 1313C NMR (100 MHz, CDCl3) δ 162.6, 156.7, 150.8, 144.6, 141.6, 139.8, 136.3, 135.8, 134.2, 132.6, 130.3, 129.9, 129.2, 128.3, 127.2, 126.9, 124.2, 118.3, 117.9, 117.0, 116.6, 112.3, 110.0, 56.9, 50.7, 30.3; C 30 H 24 Calculated m / z for C4H4N4O8S: 600.6.

[0224] Example 2. Determination of the inhibitory potency of ZAF derivatives against PDI, ERp5, ERp57 and ERp72

[0225] Two methods were used to determine the inhibitory potency of ZAF derivatives against PDI, ERp5, ERp57 and ERp72. The first assay was an insulin turbidity assay for measuring the thiol isomerase inhibitory activity of the analogs. A 30 μL mixture containing 1.75 mg / mL insulin and optimized concentrations (0.5 μg / well of PDI, ERp57 and ERp72; 1 μg of ERp5) (solvent: 100 mM K2PO4, 0.2 mM EDTA, pH 7.2) was added to a 384-well clear bottom plate. The reaction was initiated by adding 0.3 mM DTT. Absorbance at 650 nm was measured every minute. Wells without thiol isomerase, wells without inhibitor, and wells containing ZAF or MON were used as controls. The IC 50 50 of each compound was determined by running a 6-point dose curve with at least three independent replicates. This insulin inhibition assay had high nM IC 50Sensitivity limit. The second assay is a highly sensitive continuous assay in which the fluorescence of bis-(O-aminobenzoyl) glutathione disulfide (diabz-GSSG) increases upon its catalytic reduction by the above-mentioned glutathione isomerase, and is thus used to detect potent compounds. The preparation of the substrate diabz-GSSG is very simple, just by reacting isatoic anhydride and oxidized glutathione in an aqueous buffer and then performing Sephadex G-10 purification. The fluorescence of the substrate is self-quenched in the oxidized S-S form and increases significantly (about 20-fold) after enzymatic reduction. The substrate (15 mM) is pre-incubated with 50 mM DTT (where diabz-GSSH is anti-reducing), and then one of the four enzymes (5 nM - 500 nM, where the low nM concentration will be used for inhibitors with corresponding potencies) is added in 0.1 potassium phosphate buffer (pH 7.0), and the fluorescence time course (ex / em = 312 nm / 415 nm) is monitored. For each inhibitor concentration, three time courses will be collected. Then, steady-state analysis of inhibitor potency is performed using the initial velocity. The inhibition mode is determined by performing the above measurements at different substrate concentrations. Calibration is performed by measuring the signal change by completely reducing the substrate with 10 mM DTT.

[0226] ZAF and MON inhibit thiol isomerases. The potencies of ZAF and MON against PDI, ERp5, ERp57, and ERp72 were determined in an insulin turbidity assay. In the presence of dithiothreitol (DTT), insulin reduction catalyzed by thiol isomerases leads to insulin chain aggregation. The turbidity of insulin aggregation is monitored by measuring the absorbance at 650 nm. The turbidity assay based on insulin is applicable to the 384-well format. ZAF inhibits PDI, ERp5, ERp57, and ERp72, with an IC 50 value of 10 - 30 μM ( Figure 1A ); the data for MON are as Figure 1B shown.

[0227] The activities of the derived compounds as PDI and ERp57 inhibitors were tested. Derivatives 1 - 5 are compounds of scaffold I, and they all lack the cyclopentyl carbamate group at the C-5 position of the ZAF indole (A-ring). Generally, they all showed poor activities when tested as PDI and ERp57 inhibitors. Compared with the parent ZAF, the compounds belonging to scaffold IIa showed similar or higher activities as inhibitors. These data indicate that the cyclopentyl carbamate of ZAF is not essential for the activity as a thiol isomerase inhibitor, and the substituents on the B-ring maintain the 2-methoxy-5-indolyl structure unchanged, because scaffold IIa produced the most active compound 21. When testing the inhibitory activity against thiol isomerase, the derived compounds 6 - 11 did not show activity. In addition, the derived compounds 12 - 14 were found to be inactive. It can be seen that the activities of the derived compounds 15 - 20 containing N-methyl on the indole (A-ring) increased slightly, among which derivative 16 showed moderate activity (IC 50 = 30 mM) as a thiol isomerase inhibitor. The electron-withdrawing group (R 1 = NO2) on the indole (A-ring) led to an increase in thiol isomerase inhibitory activity and produced derivatives 22, 23, and 27 with moderate activity (IC 50 = 30 mM), as well as compound 28 (IC 50 = 40 mM), and the most active compound 21 (IC 50 ≈ 10 mM). By making pairwise comparisons to observe the changing trends of the substituents on the arylsulfonamide (Ar 2 ), it was found that the changing trends of the nitro and naphthalene substituents in compounds 15 - 20 (R 1 = H) and compounds 21 - 26 (R 1 = NO2) were similar, while the changing trend of the fluorine substituent was different, among which compounds 15 - 20 (3-F phenyl showed better activity (>) than 2-F phenyl > 3-NO2 phenyl ≈ 4-NO2 phenyl ≈ 2-naphthyl > 4-F) and compounds 21 - 26 (2-F phenyl > 3-F phenyl ≈ 4-F phenyl > 3-NO2 phenyl > 4-NO2 phenyl ≈ 2-naphthyl). The most active compound among the derived compounds 1 - 26 is compound 21 (Ar 2 = 2-F phenyl). To study the importance of N-methyl indole (ring A) for the most active compound 21, compounds 32 - 35 with different chain lengths were synthesized, which led to a slight decrease in the activity of the compounds (IC 50 = 50 to > 100 mM). In addition, by synthesizing compounds 27 - 31 containing other halogens and difluoro groups, the importance of 2-F phenyl for the arylsulfonamide of compound 21 was studied, which led to a slight decrease in the activity of the compounds compared with the lead compound 21.

[0228] Example 3. Toxicity Study

[0229] The cytotoxic effects of ZAF, montelukast (MON), and compounds 21, 27, and 28 on two mammalian cell lines, BEAS-2B and HEK-293, were studied. The two cell lines were seeded at 10,000 cells per well in a 96-well plate. After incubation for 24 hours, the cells were treated with 1% (positive control), 1% DMSO (negative control), and ZAF, MON, compounds 21, 27, and 28 at concentrations ranging from 0 to 62.6 mg / mL. After incubation for 12 hours, 10 μL of resazurin was added to each well, and the plate was read on a SpectraMax M5 microplate reader with a fluorescence excitation wavelength of 560 nm and an emission wavelength of 590 nm. None of the compounds showed toxicity to any of the mammalian cell lines tested (Figure 2).

[0230] Example 4. Inhibition of Cancer Cell Growth by Zafirlukast and Compound 21

[0231] ZAF and compound 21 inhibit cancer cell growth. ZAF and compound 21 were tested to evaluate the viability of HCT116 colon cancer cells and OVCAR-8 ovarian cancer cells. HEK-293 embryonic kidney cells were used as non-cancer cell controls. The IC 50 value of ZAF was in the range of 3 - 10 μM. In Figure 3, the inhibitory effect of compound 21 (inverted triangles) on cell growth was measured using a standard 3-day growth inhibition assay and compared with ZAF (circles). The potency of compound 21 was 3 to 8 times that of ZAF. ZAF and compound 21 have an inhibitory effect on mammalian cell growth. The cells were seeded and treated with the drug after 24 hours. After further incubation for 24 hours, cell viability was measured using Prestoblue. At least 3 independent experiments for each cell line are summarized in Figure 3. The cell lines used included HEK-293 non-cancer cells (not shown); OVCAR-8 and HCT116. The 3-day growth inhibition of ZAF (circles) and compound 21 (inverted triangles) on OVCAR-8 ( Figure 3A ) and HCT116 cancer cells ( Figure 3B ) was compared.

[0232] Example 5. Platelet Aggregation and Fibrin Inhibition

[0233] ZAF and MON inhibit platelet aggregation and fibrin formation. Compound 21 ( Figure 4A , Figure 4B)Inhibit platelet aggregation. In the presence of DMSO, ZAF (0.1 - 40 μM), or compound 21 (0.6 - 30 μM), platelet aggregation in washed platelet suspensions was measured and stimulated with collagen for 180 seconds. The results showed that platelet aggregation decreased in a dose - dependent manner, with the potency of compound 21 > ZAF > MON.

[0234] Thiol isomerase inhibition also affects the activation and conformational changes of integrin αIIbβ3, leading to fibrinogen ligation. The effect of ZAF on fibrinogen binding to the platelet surface was measured by flow cytometry, where ZAF pretreatment (10 μM) significantly reduced fibrinogen binding to platelets, resulting in a 50% reduction in integrin activation (2043 ± 165 AU). Higher concentrations of 20 μM ZAF further inhibited fibrinogen binding, with an inhibition rate of 61% (1633 ± 162 AU).

[0235] Example 6. ZAF inhibits factor Xa production in cancer cells

[0236] The ability of ZAF to prevent the activation of the coagulation cascade was determined by measuring its ability to inhibit factor Xa production in HCT116 cells. Cells were seeded at 50,000 cells per well in 12 - well culture dishes. After 24 hours of growth, the cells were treated with ZAF, washed twice with Tris - buffered saline (TBS), and then subjected to kinetic readings for 60 minutes in a SpectraMax with TBS containing 5 nmol / L FVIIa, 150 nmol / L FX, and 5 mmol / L CaCl2. Fluorescence was measured at excitation and emission wavelengths of 352 nm and 450 nm, respectively. We found that in the presence of ZAF, the production of factor Xa was inhibited in a dose - dependent manner ( Figure 5 ).

[0237] Example 7. ZAF inhibits platelet aggregation and thrombus formation in mice

[0238] ZAF can inhibit platelet aggregation (in vivo) and thrombosis in mice. By intravital microscopy measurement, thiol isomerase inhibition can prevent the formation of thrombosis in vivo in live mice after laser injury (see "Zafirlukast, a broad-spectrum thiol isomerase inhibitor, inhibits thrombosis without altering bleeding time" by Holbrook et al., Br. J. Pharmacol. 2021; 178: 550-563). Thiol isomerase inhibition has previously been shown to reduce fibrin formation using the same model, and it has been demonstrated that thiol isomerase inhibition can reduce tumor growth and progression without damaging normal tissues. Importantly, thiol isomerase inhibition does not increase bleeding time (Holbrook et al.), which is a major side effect of current antithrombotic therapies.

[0239] Example 8. Evaluation of the effect of compound 21 on platelet aggregation and thrombosis in mice

[0240] To evaluate the ability of compound 21 to inhibit arterial thrombosis, intravital microscopy of laser injury will be performed. Male C57BL / 6J mice, 4-5 weeks old and weighing in the range of 19-25 g, will be anesthetized by intraperitoneal injection of ketamine (125 mg / kg), xylazine (12.5 mg / kg), and atropine (0.25 mg / kg). Platelets will be labeled by intravenous infusion of DyLight 649-conjugated anti-GPIb platelet marker antibody (0.2 μg / g body weight). After exposure of the cremaster muscle of the testis, the vehicle DMSO (0.1% v / v) or a zileuton derivative will be infused intravenously, and after a 5-minute incubation period, injury to the arteriolar wall will be induced by laser ablation (Micropoint, Andor Technology, Belfast, UK). Thrombosis will then be observed using an Olympus BX microscope (Olympus, Essex, UK) and a Hamamatsu Photonics (Hamamatsu Photonics, Hertfordshire, UK) CCD camera, and data will be analyzed using Slidebook Software 5.0 (Intelligent Imaging Innovations, Denver, USA). At the end of the experiment, the mice will be killed using the method approved in Schedule 1. All animal experiments will be blinded to experimental treatment and analysis. Animal experiments have previously been approved by the University of Reading Local Ethical Review Panel and authorized by the UK Home Office. Animal research reports comply with the ARRIVE guidelines and the recommendations of the British Journal of Pharmacology.

[0241] Example 9. Ex Vivo Thrombosis

[0242] In an ex vivo thrombosis model using whole human blood, zileuton was found to inhibit thrombosis under arterial flow conditions (US20210008032A1). The active thiol isomerase inhibitor analogs described herein, such as compound 21, are expected to also inhibit ex vivo thrombosis.

[0243] Example 10. Compound 32

[0244] Compound 32 is an analogue of zileuton which, upon modification, has a significantly reduced or eliminated affinity for the leukotriene receptor 1 (LTR1) receptor, but still retains its potency as a thiol isomerase inhibitor. In an insulin turbidity assay, compound 32 maintained a potency similar to zileuton in inhibiting thiol isomerase (about 1.5-fold lower)( Figure 6 ).

[0245] Zileuton treatment also inhibited the thiol isomerase activity of OVCAR8 cells. After treatment with zileuton, the cleavage of the fluorescent eosin thiol isomerase substrate was inhibited in a concentration-dependent manner. Treatment of OVCAR8 cells with 3, 10 or 30 μM of zileuton for 10 minutes significantly inhibited PDI activity, with inhibition rates of 18%, 24% and 45% respectively( Figure 7A ). To demonstrate that the addition of zileuton did not alter the expression of thiol isomerase, the levels of thiol isomerase were measured after treating OVCAR8 cells with 10 μM and 30 μM of zileuton for 1 hour. No significant change in expression was observed( Figure 7B ), demonstrating that the observed changes in activity were due to enzyme inhibition rather than a decrease in enzyme levels. The cellular thiol isomerase activity of OVCAR8 cells was also significantly reduced after treatment with montelukast and compound 32, with the effect of montelukast being slightly lower than that of zileuton (decreased by 20%, 30% and 40% at 10, 30 and 100 μM respectively)( Figure 7C ). In contrast, compound 32 was more potent than zileuton, significantly inhibiting 25%, 39%, 29% and 44% of the activity at 1, 3, 10 and 30 μM respectively( Figure 7D ).

[0246] Since thiol isomerase activity inhibitors have been shown to have anti-tumor activity, the effects of these drugs on cancer cell viability were evaluated. Zileuton inhibited the viability of OVCAR8 cancer cells with an IC 50 of 12 μM, while montelukast and compound 32 were about 5-fold and about 1.5-fold less potent than zileuton in inhibiting cell viability (IC 50 being 60 and 20 μM respectively)( Figure 8A ). Notably, the relative potencies of zileuton, compound 32 and montelukast were consistent across various assays (cell-free assays and cell activity assays as well as between cell activity assays and viability assays), with compound 32 being similar in potency to zileuton, while montelukast was about 5-fold less potent in each assay( Figure 8B ).

[0247] Insulin-based turbidimetry

[0248] Determine the selectivity of zafirlukast and montelukast for the thiol isomerases PDI, ERp57, ERp72, and ERp5 using insulin-based turbidimetry. Dilute these drugs in a 384-well plate in a 6-12 point dose curve and add thiol isomerases at a final concentration of 10 μg / mL (30 μg / mL for ERp5 only), 125 μM insulin, 2 mM EDTA, and 100 mM potassium phosphate buffer, with a total volume of 30 μL per well. After initiating the reaction with 0.3 mM DTT, measure the turbidity of insulin aggregation once per minute for 75 minutes using a SpectraMax M3 microplate reader (Molecular Devices, Sunnyvale, CA).

[0249] PrestoBlue assay

[0250] Seed the cell line at 5,000 cells per well in a 96-well plate and allow it to grow for 24 hours. Then re-treat the cells with the drug or vehicle control for 2 - 24 hours, and then add PrestoBlue reagent (Invitrogen, Waltham, MA) and incubate at 37 °C for 10 - 20 minutes. Determine cell viability by measuring the fluorescence signal at an excitation wavelength of 560 nm and an emission wavelength of 590 nm. Normalize the signal to a percentage of the control.

[0251] Eosin-GSSG disulfide reductase assay

[0252] Seed OVCAR8 cells at 10,000 cells per well, allow them to grow overnight, and then treat them with 0 - 100 μM zafirlukast, montelukast, or compound 32 for 10 minutes. Then detect the samples with 150 nM eosin-GSSG probe in the presence of 5 μM DTT and phosphate buffer 1 (containing 100 mM potassium phosphate (pH 7.4) and 2 mM EDTA). Monitor the increase in fluorescence every 30 seconds for 30 minutes by exciting at 520 nm and emitting at 550 nm. Then normalize the generated data for each sample to the control, and the raw data represent relative fluorescence units (RFU) / minute.

[0253] For blood samples, assays were performed as previously described (Raturi, A. and Mutus, B. (2007) Characterization of the redox state and reductase activity of 24 protein disulfide isomerases in different redox environments using a sensitive 25 fluorometric assay. Free Radic Biol Med 43, 62-70), with modifications, namely using plasma diluted 1:1 with potassium phosphate buffer (containing 100 mM potassium phosphate (pH 7.4) and 2 mM EDTA). Samples were then assayed with 150 nM eosin-GSSG probe in the presence of 5 μM DTT. The increase in fluorescence was monitored by excitation at 520 nm and emission at 550 nm for 30 minutes. Data generated on day 28 were then normalized for each sample against the control on day 0, and the raw data represent relative fluorescence units (RFU) / minute (n = 3 for each patient sample).

[0254] Statistical analyses were performed using GraphPad Prism (version 9.4.0, San Diego, CA). Data are presented as mean ± SD. For the eosin-GSSG assay, statistical analysis was performed between experimental and control groups using one-way ANOVA and post-hoc Dunnett’s test. *P < 0.05, **P < 0.01, ***P < 0.001 or ****P < 0.0001 was considered statistically significant.

[0255] Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0256] Unless the context clearly dictates otherwise, the singular forms “a” and “the” include plural referents. All ranges of endpoints for the same feature or component are combinable independently and include the recited endpoints. As used herein, the plural suffix “(s)” is intended to include both the singular and plural of the term it modifies, thereby including one or more of that term (e.g., carrier includes one or more carriers). Unless the context clearly dictates otherwise, the term “or” means “and / or”. The term “combination” includes blends, mixtures, and the like.

[0257] References to “one embodiment,” “another embodiment,” “some embodiments,” etc., throughout the specification mean that a particular element (e.g., feature, structure, step, or property) associated with the embodiment is included in at least one embodiment described herein, and may or may not be present in other embodiments. Additionally, it should be understood that the described elements may be combined in any suitable manner in various embodiments.

[0258] In general, a composition, method, and article of manufacture can alternatively include any ingredient, step, or component disclosed herein, consist of any ingredient, step, or component disclosed herein, or consist essentially of any ingredient, step, or component disclosed herein. A composition, method, and article of manufacture can additionally or alternatively be formulated, practiced, or manufactured so as to be free, or substantially free, of any ingredient, step, or component that is not necessary to achieve the functions or objectives of the present claims.

[0259] "Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes instances where the event occurs and instances where it does not.

[0260] As used herein, "about" or "approximately" includes the recited value and means within an acceptable deviation range of the recited value as determined by one of ordinary skill in the art in view of the relevant measurements and the error associated with the measurement of a particular quantity (e.g., the limitations of the measurement system). For example, "about" can mean within one or more standard deviations.

[0261] Although the present invention has been described in detail in connection with only a limited number of embodiments, it should be readily understood that the invention is not limited to these disclosed embodiments. Rather, the invention can be modified to incorporate any number of variations, alterations, substitutions, or equivalent arrangements that have not been previously described but are within the spirit and scope of the invention. Additionally, although various embodiments of the invention have been described, it should be understood that aspects of the invention can include only some of the described embodiments. Accordingly, the invention is not to be considered as limited by the foregoing description, but is only limited by the scope of the appended claims.

[0262] The entire contents of all cited patents, patent applications, and other references are incorporated herein by reference. However, if a term in this application contradicts or conflicts with a term in the incorporated references, the term in this application shall control over the conflicting term in the incorporated reference.

Claims

1. A method for treating cancer or treating or preventing cancer-induced thrombosis, wherein, The method comprises: administering to a patient in need thereof a therapeutically effective amount of a compound of formula (I), (II), (III) or a pharmaceutically acceptable salt thereof: wherein, Ar 1 For R 1 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxy, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 , wherein R 9 is hydrogen, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH or aryl; Q 1 and Q 2 are each independently a bond, O or NR 10 , and R 10 is hydrogen, C1-C8 alkyl, C1-C6 haloalkyl or C1-C8 alkyl-OH; R 2 is hydrogen, a C1-C8 alkyl group, a C1-C6 haloalkyl group, a C1-C8 alkyl-OH group, a C3-C7 cycloalkyl group, a (C3-C7 cycloalkyl)C0-C6 alkyl group, a C1-C4 alkanoyl group, or an unsubstituted or substituted aryl group; X is O, S or N; Y is N or CH; When X is N, R 6 is hydrogen, C1-C8 alkyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl or C1-C4 alkanoyl, and when X is S or O, R does not exist 6 ; R 7 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxyl, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 where Q 1 、Q 2 and R 9 are as defined above; Ar 2 is unsubstituted or substituted by R 8 substituted aryl or unsubstituted or substituted by R 8 substituted heteroaryl, wherein R 8 is NO2, cyano, halogen, NH2, COOH, hydroxy, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 , wherein Q 1 、Q 2 and R 9 are as defined above; R 4 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxy, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 , where Q 1 、Q 2 and R 9 are as defined above; and R 5 is hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl or C1-C4 alkanoyl, subject to the following conditions a) and b): a) When Ar 1 is R 4 is -OMe, R 5 is hydrogen and Ar 2 is phenyl or phenyl substituted by R 8 for the compound of formula (III), R 1 , R 2 and R 8 do not satisfy the following six conditions: b) When Ar 1 is R 1 is hydrogen, NO2, NH2, mono-C1-C8 alkylamino or di-C1-C8 alkylamino; R 2 is hydrogen or C1-C8 alkyl; R 4 is hydrogen or C1-C8 alkyl; and when R 5 is hydrogen; then Ar 2 is not unsubstituted phenyl or phenyl substituted with a C1-C8 alkyl group at the 2-position.

2. The method according to claim 1, wherein, Ar 1 For 3. The method according to any one of claims 1 to 2, wherein, R 1 is an electron-withdrawing group; or R 1 is NO2, cyano, C1-C6 haloalkyl, CF3 or C2-C6 alkanoyl.

4. The method according to any one of claims 1 to 3, wherein, R 2 is hydrogen, a C1-C6 alkyl group, a C1-C8 alkyl-OH group, or an unsubstituted or substituted phenyl group.

5. The method according to claim 1, wherein, Ar 1 For X is O, S or N; Y is N or CH; and When X is N, R 6 is hydrogen, C1-C8 alkyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl or C1-C4 alkanoyl, and when X is S or O, R does not exist 6 .

6. The method according to claim 1, wherein, Ar 1 For And R 7 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxyl, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 , where Q 1 、Q 2 and R 9 are as defined above.

7. The method according to any one of claims 1 to 6, wherein, Ar 2 is an unsubstituted phenyl or naphthyl, or a phenyl or naphthyl substituted by R 8 .

8. The method according to any one of claims 1 to 6, wherein, Ar 2 is a halogen-substituted phenyl group or 2-naphthyl.

9. The method according to any one of claims 1 to 8, wherein, R 4 is a C1-C6 alkyl group or a C1-C6 alkoxy group.

10. The method according to any one of claims 1 to 8, wherein R 4 is a C1-C2 alkyl or C1-C2 alkoxy group.

11. The method according to any one of claims 1 to 10, wherein R 5 is hydrogen.

12. The method according to any one of claims 1 to 11, wherein the cancer is breast cancer, colon cancer, colorectal cancer, glioma, hematological cancer, laryngeal cancer, lung cancer, lymphoma, melanoma, neuroblastoma, ovarian cancer, prostate cancer or a combination thereof.

13. The method according to any one of claims 1 to 11, wherein The cancer-induced thrombosis is arterial thrombosis, venous thrombosis or a combination thereof.

14. The method according to any one of claims 1 to 13, wherein The method further comprises providing the patient with an additional pharmaceutically active ingredient.

15. The method according to claim 14, wherein The additional pharmaceutically active ingredient is an antithrombotic agent, an anticoagulant, a chemotherapeutic agent, an antiviral agent or an anti-inflammatory agent.

16. A method for preventing or treating thrombosis, thrombotic diseases, platelet aggregation, fibrin formation, infectious diseases, viral diseases, immune diseases, inflammation, neurological diseases, neurodegenerative diseases or a combination thereof in a patient, wherein The method comprises: administering to the patient in need thereof a therapeutically effective amount of a compound of formula (I), (II), (III) or a pharmaceutically acceptable salt thereof.

17. The method according to claim 16, wherein The thrombosis is arterial thrombosis or venous thrombosis, and wherein the thrombotic disease is acute myocardial infarction, stable angina, unstable angina, acute occlusion after coronary angioplasty and / or stent implantation, transient ischemic attack, cerebrovascular disease, stroke, peripheral vascular disease, placental insufficiency, atrial fibrillation, deep vein thrombosis and pulmonary embolism or a combination thereof.

18. A compound of formula (I), (II), (III) or a pharmaceutically acceptable salt thereof: wherein Ar 1 For R 1 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxyl, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 , wherein R 9 is hydrogen, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH or aryl; Q 1 and Q 2 are each independently a bond, O or NR 10 , and R 10 is hydrogen, C1-C8 alkyl, C1-C6 haloalkyl or C1-C8 alkyl-OH; R 2 is hydrogen, a C1-C8 alkyl group, a C1-C6 haloalkyl group, a C1-C8 alkyl-OH group, a C3-C7 cycloalkyl group, a (C3-C7 cycloalkyl)C0-C6 alkyl group, a C1-C4 alkanoyl group, or an unsubstituted or substituted aryl group; X is O, S or N; Y is N or CH; When X is N, R 6 is hydrogen, C1-C8 alkyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl or C1-C4 alkanoyl, and when X is S or O, R does not exist 6 ; R 7 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxyl, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 , wherein Q 1 、Q 2 and R 9 are as defined above; Ar 2 is unsubstituted or substituted aryl or unsubstituted or substituted heteroaryl; R 4 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxyl, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 , where Q 1 、Q 2 and R 9 are as defined above; and R 5 is hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl or C1-C4 alkanoyl, subject to the following conditions a) and b): a) When Ar 1 is R 4 is -OMe, R 5 is hydrogen and Ar 2 is phenyl or phenyl substituted by R 8 for the compound of formula (III), R 1 , R 2 and R 8 do not satisfy the following six conditions: b) When Ar 1 is R 1 is hydrogen, NO2, NH2, mono-C1-C8 alkylamino or di-C1-C8 alkylamino; R 2 is hydrogen or C1-C8 alkyl; R 4 is hydrogen or C1-C8 alkyl; and when R 5 is hydrogen, Ar 2 is not unsubstituted phenyl or phenyl substituted with a C1-C8 alkyl group at the 2-position.

19. The compound according to claim 18, wherein Ar 1 for 20. The compound according to claim 18 or 19, wherein, R 1 is an electron-withdrawing group; or R 1 is NO2, cyano, C1-C6 haloalkyl, CF3 or C2-C6 alkanoyl.

21. The compound according to any one of claims 18 to 20, wherein, R 2 is hydrogen, C1-C6 alkyl, C1-C8 alkyl-OH or unsubstituted or substituted phenyl.

22. The compound according to claim 18, wherein, Ar 1 For X is O, S or N; Y is N or CH; and When X is N, R 6 is hydrogen, C1-C8 alkyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl or C1-C4 alkanoyl; when X is S or O, R does not exist 6 .

23. The compound according to claim 18, wherein, Ar 1 is and R 7 is hydrogen, NO2, cyano, halogen, NH2, COOH, hydroxyl, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 , where Q 1 , Q 2 and R 9 are as defined above.

24. The compound according to any one of claims 18 to 23, wherein, Ar 2 is unsubstituted or substituted by R 8 to form a phenyl or naphthalene, where R 8 is NO2, cyano, halogen, NH2, COOH, hydroxy, C1-C8 alkyl, C1-C6 haloalkyl, C1-C8 alkyl-OH, C1-C8 alkyl-NH2, C1-C8 alkoxy, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C2-C6 alkenyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C0-C6 alkyl, C2-C6 alkanoyl or -Q 1 (C=O)Q 2 R 9 where Q 1 and Q 2 and R 9 are as defined above.

25. The compound according to any one of claims 18 to 23, wherein, Ar 2 is a halogen-substituted phenyl group or 2-naphthyl.

26. The compound according to any one of claims 18 to 25, wherein, R 4 is a C1-C6 alkyl or a C1-C6 alkoxy group.

27. The compound according to any one of claims 18 to 25, wherein, R 4 is a C1-C2 alkyl or C1-C2 alkoxy group.

28. The compound according to any one of claims 18 to 27, wherein, R 5 is hydrogen.

29. A pharmaceutical composition, wherein, the pharmaceutical composition comprises a compound according to any one of claims 18 to 28 and a pharmaceutically acceptable excipient.

30. The pharmaceutical composition according to claim 29, wherein, The pharmaceutical composition is formulated for oral, topical, parenteral, by inhalation or spray, sublingual, transdermal, buccal or rectal administration.

31. The pharmaceutical composition according to claim 29, wherein, The pharmaceutical composition is formulated as a tablet or a capsule.

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