Methods of Treatment with BCN057, BCN077 and Analogs

Abstract

The present disclosure relates to methods of treating or alleviating various conditions by administration of BCN057, BCN077 and analogs. BCN057, BCN077 and analogs can be used to modulate PD-1 and treat various diseases in which PD-1 expression is implicated. The compounds have anti-neoplastic activity. In addition, the compounds also have cytoprotective activity as they protect against chemotherapy-induced toxicity to the GI tract. The compounds disclosed herein can be used to reduce tumor burden in cancers, including pancreatic, gastrointestinal (GI) and epithelial cancers.

Description

[Technical field]

[0001] Related Applications This application claims priority to U.S. Provisional Application No. 63 / 240,847, filed September 3, 2021, the contents of which are incorporated herein by reference.

[0002] FIELD OF THEINVENTION The present invention relates to small molecules for therapeutic use, and more particularly to the therapeutic applications of BCN057, BCN077 and analogues thereof. [Background technology]

[0003] Cancer can be defined as a group of diseases involving abnormal cell growth that invades or spreads to other parts of the body. Despite technological advances, cancer continues to be a major cause of death and immense suffering. Cancer is the second most common cause of death in the United States.

[0004] Cancer patients often have limited treatment options. Treatments may include a combination of surgery, radiation therapy, chemotherapy, and targeted therapy. Despite advances in research, these treatments have remained relatively unchanged in recent decades. Efforts are often focused on early diagnosis of cancer, when treatments are more effective. However, early diagnosis offers only a small margin of survival.

[0005] Pancreatic cancer occurs when cells in the pancreas, a glandular organ located behind the stomach, begin to grow uncontrollably and form a mass. These cancer cells have the ability to invade other parts of the body. Many types of pancreatic cancer are known. However, the most common type, pancreatic adenocarcinoma, accounts for about 90% of cases. Signs and symptoms of the most common form of pancreatic cancer include yellow skin, abdominal or stomach pain, unexplained weight loss, pale stools, dark urine, and loss of appetite. Symptoms are usually not observed in the early stages of the disease. Symptoms specific enough to suggest pancreatic cancer do not usually occur until the disease has reached an advanced stage. By the time of diagnosis, pancreatic cancer has often spread to other parts of the body.

[0006] Chemotherapy is a type of cancer treatment that uses one or more anti-cancer drugs (chemotherapeutic agents) as part of a standard chemotherapy regimen. Conventional chemotherapy agents are cytotoxic by interfering with cell division. However, cancer cells vary in their sensitivity to these agents. In general, chemotherapy can be considered to lead to cell damage or stress, which can then lead to death when apoptosis is initiated. Many side effects of chemotherapy can be traced back to damage to normal cells that divide rapidly and are therefore sensitive to antimitotic drugs. This results in the most common chemotherapy side effects: bone marrow suppression, mucositis, and alopecia. Due to their effects on immune cells (especially lymphocytes), chemotherapy drugs are often used in a host of diseases that result from a harmful overactivity of the immune system against the self (so-called autoimmunity).

[0007] Checkpoint inhibitor receptors expressed on immune cells (e.g., CTLA4 and PD-1) activate immune-suppressive signaling pathways. CTLA4 competes with CD28 for binding to CD80 and CD86. PD-1 binds to PD-L1 or PD-L2 and blocks positive signals through the T cell receptor (TCR) and CD28. These immune-suppressive molecules act as brakes to regulate adaptive immune responses. Inhibitory signals can be used in a variety of ways to maintain the balance of the immune system. Based on this application, the PD-1 / PD-L pathway stands out as an immune checkpoint. In general, expression of PD-L1 is observed on T, B, and antigen-presenting cells, as well as in some non-lymphoid tissues.

[0008] Identification of tumor-specific antigens, lymphocytes, and T-cell responses in cancer patients leads to immunotherapies that are designed to boost anti-tumor immune responses. Biologically derived substances, or "biologics," include recombinant therapeutic proteins such as monoclonal antibodies (MABs). Biological drugs can stimulate the body to find and attack cancer cells. Some biologics attack cancer cells directly and disrupt their growth signals. Other biologics can help patients fight infections after chemotherapy.

[0009] T cells are powerful cellular effectors of the immune system and have memory to respond when rechallenged by the same antigen. Tumor immunotherapy is more specific and less toxic than traditional chemotherapy agents, so it may be an ideal adjuvant therapy for patients with malignant tumors at risk of recurrence. Chemotherapy is used first to induce active disease into remission, followed by tumor immunotherapy via memory T cells. However, biologics also have limitations. Although immunotherapies appear to be effective against small tumor burdens, they cannot control large tumor masses. In addition, patients with advanced disease have damaged immunity, which limits the effectiveness of immunotherapy. Therefore, it has been proposed that immunotherapy should be used at an early stage of tumor formation. Moreover, biologics are expensive compared to other drugs, and they need to be administered intravenously. Furthermore, their relatively large size limits their function and half-life in the blood.

[0010] Recent research has provided new opportunities for cancer therapy, including the use of small molecules. Certain small molecules bind biological micromolecules and function as effectors, thereby altering the activity or function of the target. They manifest various biological functions or applications and function as intracellular signaling molecules. For example, small molecule cancer drugs have been successfully used to target extracellular, cell surface receptors as well as intracellular proteins, including anti-apoptotic proteins that play important roles in downstream signaling to promote cell proliferation and metastasis. Many small molecule drugs inhibit important cancer targets such as serine / tyrosine kinases, matrix metalloproteinases (MMPs), heat shock proteins (HSPs), proteosomes and other proteins important in signaling pathways. The small size (<500 Da) of these molecules also allows them to translocate through the cell membrane. In principle, small molecule compounds can be developed to target any part of the cell, regardless of the subcellular location of the target. Such drugs have good therapeutic potential because they can inhibit important cancer targets.

[0011] Effective treatment for pancreatic cancer is challenging due to resistance to radiation / chemotherapy and poor drug permeability. In addition, treatment-induced normal tissue toxicity, mainly to the duodenal and gastrointestinal epithelium, is a common and dose-limiting adverse event. Gastrointestinal toxicity often results in the interruption, reduction or early withdrawal of anticancer therapy, a significant factor affecting the overall survival of treated patients. Thus, there is a pressing need to develop therapeutic strategies that selectively sensitize tumor tissues without inducing normal tissue toxicity. An ideal treatment / therapy should target cancer cells and reduce the deleterious effects of chemotherapy. Summary of the Invention

[0012] The compound BCN057 disclosed herein was previously described in US Patent Application No. 13 / 813,923 and US Patent Application No. 14 / 889,719. The present invention provides novel molecules and analogs and novel methods of use. BCN057 can also be referred to by its structural name: (3-[(furan-2-ylmethyl)-amino]-2-(7-methoxy-2-oxo-1,2-dihydroquinolin-3-yl)-6-methyl-imidazo[1,2-a]pyridin-1-ium) or YEL002. BCN057 and its analogs are useful for treating or preventing inflammatory diseases and for treating or preventing cancer or other hyperproliferative conditions, among other therapies. [ka]

[0013] Applicants propose that the small molecules BCN057, BCN077 can modulate the chemosensitivity of oncogenic RAS pancreatic cancer cells, but conversely protect normal intestinal epithelium from off-target toxicity from these small molecules by alleviating chemotoxicity in Lgr5-positive intestinal stem cells, thereby protecting barrier function. Furthermore, it is shown that BCN057 and BCN077 inhibit GSK3β, thereby inducing a proapoptotic phosphorylation pattern for c-Jun in KRAS G12D mutant pancreatic cancer cells, leading to restoration of PTEN expression and resulting apoptosis, which is a novel mechanistic insight into the oncogenic RAS phenotype. Finally, in conjunction with their GSK3β inhibition, BCN057 and BCN077 are small molecule inhibitors of PD-1 expression on human T lymphocytes co-cultured with human pancreatic cancer cells. Taken together, BCN057 and BCN077 are able to promote synthetic lethality specifically against malignant cells and should therefore be considered to improve the therapeutic ratio in pancreatic and epithelial cancer treatment in conjunction with chemotherapy and radiotherapy.

[0014] One embodiment is a compound of formula I: [ka] or an analogue thereof, R 1 = alkyl, alkylamine, ether, aryl or aryl halide, R 2 =H or CH3, R 3 =H, OH or OCH3. Or, R 1 = [ka] and n=1, 2 or 3; X=F, CF3Cl, OH or CH2OH; Y=O, S or N; Z=CH3, CH2OH or OCH3. Or, R 1 = [ka] where n=1, 2 or 3; X=F, CF3Cl, OH or CH2OH; Y=O, S or N; and Z=CH3, CH2OH or OCH3. Or, R 1 = [ka] where n=1, 2 or 3; X=F, CF3Cl, OH or CH2OH; and Y=O, S or N. Or, R 1 = [ka] where n=1, 2 or 3.

[0015] One embodiment is a method of treating cancer comprising administering a therapeutic amount of a compound or analog thereof of Formula I. The cancer can be, for example, pancreatic, brain, breast, colorectal, cervical, gastrointestinal, genitourinary, head and neck, lung, ovarian, pancreatic, prostate, renal, rectal, skin, blood, or testicular cancer.

[0016] The cancer may also be biliary tract cancer, bladder cancer, ganglionic carcinoma (neuroblastoma), leukemia, lymphoma, liver cancer (e.g., hepatocellular carcinoma), lung cancer (e.g., large cell carcinoma, non-small cell carcinoma, and squamous cell carcinoma), soft tissue cancer (e.g., angiosarcoma, leiomyosarcoma, liposarcoma, rhabdomyosarcoma, myxoma, and malignant fibrous histiocytoma-malignant fibrous histiocytoma), gastric cancer, or thyroid cancer.

[0017] Another embodiment is a method of modulating PD-1 expression to treat a disease comprising administering a therapeutic amount of a compound of Formula I or an analog thereof. The disease can be, for example, cancer, cerebral malaria, Trypanosoma cruzi induced myocarditis, influenza virus A, tuberculosis, chlamydia pulmonary infection, COPD, acute lung injury, HBV & HCV liver infection, pancreatitis, type 1 diabetes, sepsis, or HIV-1.

[0018] Another embodiment is a method of inhibiting PD-1, PD-L1 and / or PD-1 / PD-L1 interaction comprising administering an effective amount of a compound of formula I or an analog thereof. In some aspects, the method blocks the interaction of PD-L1 with PD-1 and / or CD80. The method may include administration of an additional therapeutic agent, such as a small molecule, an antibody, an antibody fragment, an antibody conjugate, or an immunomodulatory agent. In some aspects, the method includes administration of one or more immune checkpoint regulators.

[0019] Additional embodiments include methods of treating a viral infection in a subject in need thereof with a compound of formula I or an analog thereof. Some embodiments include methods of treating depression with a compound of formula I or an analog thereof.

[0020] Additional embodiments include methods of treating a viral infection in a subject in need thereof with a compound of formula I or an analog thereof. Some embodiments include methods of treating depression with a compound of formula I or an analog thereof.

[0021] One embodiment is a compound of formula A. Additional embodiments include a method of treating a disease in a subject comprising administering to the subject a therapeutically effective amount of a compound having the structure of formula A below, or an analog thereof. [ka] [ka]

[0022] Additional embodiments include a method of treating cancer in a subject with a therapeutically effective amount of a compound of formula A or an analog thereof. The cancer can be pancreatic, brain, breast, colorectal, cervical, gastrointestinal, genitourinary, head and neck, lung, ovarian, pancreatic, prostate, renal, rectal, skin, blood or testicular cancer. Treatment can include administering a chemotherapeutic or radiotherapeutic agent to the subject, as well as one or more additional agents.

[0023] Additional embodiments include methods of treating a subject having one or more side effects of a chemotherapeutic or radiotherapeutic agent with a compound of formula A or an analog thereof. Yet another embodiment is a method of preventing or treating radiation-induced damage to epithelial cells by administering a therapeutically effective amount of a compound of formula B or an analog thereof. Radiation-induced damage to epithelial cells may be identified as radiation-induced gastrointestinal syndrome (RIGS), radiation-induced mucositis, radiation-induced oral mucositis, radiation-induced proctitis and / or radiation-induced enteritis.

[0024] Additional embodiments include methods of treating fibrosis with a compound of Formula A or an analog thereof. The fibrosis can be pulmonary fibrosis, idiopathic pulmonary fibrosis, acute respiratory distress syndrome, cystic fibrosis, non-cystic fibrosis bronchiectasis, liver cirrhosis, hepatic fibrosis, endomyocardial fibrosis, old myocardial infarction, atrial fibrosis, mediastinal fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis, nephrogenic systemic fibrosis, Crohn's disease, gastrointestinal fibrosis, keloid conditions, scleroderma / systemic sclerosis, arthrofibrosis, Peyronie's disease, Dupuytren's contracture, oral submucous fibrosis, hepatic fibrosis, gastrointestinal fibrosis, dialysis-derived renal fibrosis, and / or adhesive capsulitis.

[0025] Additional embodiments include methods of treating a viral infection in a subject in need thereof with a compound of formula A or an analog thereof. Some embodiments include methods of treating depression with a compound of formula B or an analog thereof.

[0026] Another embodiment is a method of inhibiting PD-1, PD-L1 and / or PD-1 / PD-L1 interaction comprising administering an effective amount of a compound of formula A or an analog thereof. In some aspects, the method blocks the interaction of PD-L1 with PD-1 and / or CD80. The method may include administration of an additional therapeutic agent, such as a small molecule, an antibody, an antibody fragment, an antibody conjugate, or an immunomodulatory agent. In some aspects, the method includes administration of one or more immune checkpoint regulators.

[0027] Another embodiment is a compound of formula B: [ka] or an analog thereof, R 1 is an alkyl, alkylamine, or ether; R 2 is H or CH3; and R 3 is H or OH. Analogs of Formula B can include those detailed in Table 2 below.

[0028] Additional embodiments include a method of treating cancer in a subject with a therapeutically effective amount of a compound of formula B or an analog thereof. The cancer can be pancreatic, brain, breast, colorectal, cervical, gastrointestinal, genitourinary, head and neck, lung, ovarian, pancreatic, prostate, renal, rectal, skin, blood or testicular cancer. Treatment can include administering a chemotherapeutic or radiotherapeutic agent to the subject, as well as one or more additional agents.

[0029] Additional embodiments include methods of treating a subject having one or more side effects of a chemotherapeutic or radiotherapeutic agent with a compound of formula B or an analog thereof. Another embodiment is a method of preventing or treating radiation-induced damage to epithelial cells by administering a therapeutically effective amount of a compound of formula B or an analog thereof. Radiation-induced damage to epithelial cells may be identified as radiation-induced gastrointestinal syndrome (RIGS), radiation-induced mucositis, radiation-induced oral mucositis, radiation-induced proctitis and / or radiation-induced enteritis.

[0030] Additional embodiments include methods of treating fibrosis with a compound of formula B or an analog thereof. The fibrosis can be pulmonary fibrosis, idiopathic pulmonary fibrosis, acute respiratory distress syndrome, cystic fibrosis, non-cystic fibrosis bronchiectasis, liver cirrhosis, hepatic fibrosis, endomyocardial fibrosis, old myocardial infarction, atrial fibrosis, mediastinal fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis, nephrogenic systemic fibrosis, Crohn's disease, gastrointestinal fibrosis, keloid conditions, scleroderma / systemic sclerosis, arthrofibrosis, Peyronie's disease, Dupuytren's contracture, oral submucous fibrosis, hepatic fibrosis, gastrointestinal fibrosis, dialysis-derived renal fibrosis, and / or adhesive capsulitis.

[0031] Additional embodiments include methods of treating a viral infection in a subject in need thereof with a compound of formula B or an analog thereof. Some embodiments include methods of treating depression with a compound of formula B or an analog thereof.

[0032] Another embodiment is a method of treating a disease in a subject comprising administering to the subject a therapeutically effective amount of a compound having the structure of Formula C: [ka] R 3 = [ka] and Y=OCH2CH3, OCH(n)CH3, NH2, OH, OMe, Me, H, CH2OH, BH2, SeMe or SMe; R 1 and R 2 are independently hydrogen, a straight or branched C1-C20 alkyl, alkenyl, alkynyl, which may be substituted or unsubstituted, a cycloalkyl, a cycloalkenyl, a heterocyclic alkyl, or a heterocyclic alkenyl, which may be substituted or unsubstituted, a phenyl, a substituted phenyl, an aryl, a substituted aryl, an amino, an amido, F, Cl, Br, I, nitro, hydroxyl, thiol, an alkylthio, a selenol, an alkylselenyl, a silyl, a siloxy, a boryl, a carboxylic acid, a sulfolyl, -SO4H, an alkoxy, or an acyl group.

[0033] One embodiment is a method of treating cancer comprising administering a therapeutic amount of a compound of formula C or an analog thereof. The cancer can be, for example, pancreatic, brain, breast, colorectal, cervical, gastrointestinal, genitourinary, head and neck, lung, ovarian, pancreatic, prostate, renal, rectal, skin, blood or testicular cancer.

[0034] The cancer may also be biliary tract cancer, bladder cancer, ganglionic carcinoma (neuroblastoma), leukemia, lymphoma, liver cancer (e.g., hepatocellular carcinoma), lung cancer (e.g., large cell carcinoma, non-small cell carcinoma, and squamous cell carcinoma), soft tissue cancer (e.g., angiosarcoma, leiomyosarcoma, liposarcoma, rhabdomyosarcoma, myxoma, and malignant fibrous histiocytoma-malignant fibrous histiocytoma), gastric cancer, or thyroid cancer.

[0035] Another embodiment is a method of modulating PD-1 expression to treat a disease comprising administering a therapeutic amount of a compound of Formula C or an analog thereof. The disease can be, for example, cancer, cerebral malaria, Trypanosoma cruzi induced myocarditis, influenza virus A, tuberculosis, chlamydia lung infection, COPD, acute lung injury, HBV & HCV liver infection, pancreatitis, type 1 diabetes, sepsis, or HIV-1.

[0036] Additional embodiments include methods of treating a viral infection in a subject in need thereof with a compound of formula C or an analog thereof. Some embodiments include methods of treating depression with a compound of formula C or an analog thereof.

[0037] Additional embodiments include methods of treating a viral infection in a subject in need thereof with a compound of formula C or an analog thereof. Some embodiments include methods of treating depression with a compound of formula I or an analog thereof.

[0038] Another embodiment is a method of inhibiting PD-1, PD-L1 and / or PD-1 / PD-L1 interaction comprising administering an effective amount of a compound of formula C or an analog thereof. In some aspects, the method blocks the interaction of PD-L1 with PD-1 and / or CD80. The method may include administration of an additional therapeutic agent, such as a small molecule, an antibody, an antibody fragment, an antibody conjugate, or an immunomodulatory agent. In some aspects, the method includes administration of one or more immune checkpoint regulators.

[0039] In another embodiment, the disclosure provides a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the compounds of BCN057 and BCN077, or analogs thereof.In another embodiment, the disclosure provides a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a drug combination comprising a therapeutically effective amount of the compounds of BCN057 and BCN077, or analogs thereof.

[0040] In another embodiment, the disclosure provides a method of treating a side effect of chemotherapy or radiation therapy in a subject, the method comprising administering to the subject a therapeutically effective amount of the compounds BCN057 and BCN077, or analogs thereof.In another embodiment, the disclosure provides a method of treating radiation-induced gastrointestinal syndrome (RIGS) in a subject, the method comprising administering to the subject a therapeutically effective amount of the compounds BCN057 and BCN077, or analogs thereof.

[0041] Another embodiment is a method of blocking PD-L1 interaction with PD-1 and / or CD80 in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of formula C, or an analog thereof.

[0042] In certain embodiments, the compounds of the present invention may be prodrugs of compounds of formula I or II, for example, a hydroxyl in the parent compound is provided as an ester or carbonate, or a carboxylic acid present in the parent compound is provided as an ester. In certain such embodiments, the prodrug is metabolized in vivo to the active parent compound (e.g., an ester is hydrolyzed to the corresponding hydroxyl or carboxylic acid).

[0043] In certain embodiments, the compounds of the present invention may be racemic. In certain embodiments, the compounds of the present invention may be enriched in one enantiomer. For example, the compounds of the present invention may have more than 30% ee, more than 40% ee, more than 50% ee, more than 60% ee, more than 70% ee, more than 80% ee, more than 90% ee, or more than 95% ee. In certain embodiments, the compounds of the present invention may have two or more stereocenters. In certain embodiments, the compounds of the present invention may be enriched in one or more diastereomers. For example, the compounds of the present invention may have more than 30% de, more than 40% de, more than 50% de, more than 60% de, more than 70% de, more than 80% de, more than 90% de, or more than 95% de.

[0044] In certain embodiments, the present invention relates to methods of treatment with the above compounds, their analogs and / or pharma- ceutically acceptable salts, hi certain embodiments, the therapeutic formulations may be enriched to provide predominantly one enantiomer of the compound.

[0045] In certain embodiments, a therapeutic formulation may be enriched to provide predominantly one diastereomer of a compound. A diastereomerically enriched mixture may contain, for example, at least 60 mole percent, or more preferably at least 75, 90, 95, or 99 mole percent of one diastereomer.

[0046] In certain embodiments, the present invention relates to methods of treatment with Formula I, A, B or C, or pharma- ceutically acceptable salts thereof, hi certain embodiments, the therapeutic formulation may be enriched to provide predominantly one enantiomer of the compound.

[0047] In certain embodiments, the therapeutic formulation may be concentrated to provide mainly one diastereomer of the compound. In certain embodiments, the present invention provides a pharmaceutical formulation suitable for use in a human patient, comprising any of the compounds described above and one or more pharma- ceutically acceptable excipients. In certain embodiments, the pharmaceutical formulation may be for use in treating or preventing a condition or disease described herein. In certain embodiments, the pharmaceutical formulation has sufficiently low pyrogen activity so as to be suitable for use in a human patient. Any of the compounds of the above structure may be used in the manufacture of a medicament for the treatment of any of the diseases or conditions disclosed herein.

[0048] Various other objects, features and attendant advantages of the present invention will be more fully appreciated as the same becomes better understood when considered in conjunction with the accompanying drawings, in which like reference characters indicate the same or similar parts throughout the several views. [Brief description of the drawings]

[0049] [Figure 1] 1 is a histogram showing the effect of BCN057 on pancreatic cancer cell survival. [Figure 2A] FIG. 11 is a panel of images showing pancreatic cancer cell proliferation at 0, 15, 30, 60, 120, and 240 minutes after treatment with 10 μM BCN057. [Figure 2B] 1 is a line graph showing the luminescence (RLU) of Panc-1 cells treated with serially diluted BCN057. [Figure 2C] FIG. 1 is a histogram showing the relative expression of qPCR analysis of pro- and anti-apoptotic markers: Bax, caspase-3, Akt-1, and BCl-2 in Panc-1 cells. [Figure 3A] FIG. 1 is a Western blot showing increased GSK3β phosphorylation at S-9 following addition of 10 μM BCN057 in Panc-1 cells. [Figure 3B] 1 is a histogram showing relative expression as the ratio of phosphorylated GSK3B / total GSK3B protein. [Figure 4A] Western blot showing that BCN057 increases JNK-dependent phosphorylation in KRAS mutant Panc-1 cancer cells after drug treatment. Phosphorylation of c-Jun at T-91 / 93 is known to induce apoptosis. Total c-Jun protein levels were unchanged. [Figure 4B] Western blot of PTEN protein expression over 24 hours with β-actin staining from the same samples with vehicle or BCN057 treatment (10 uM) at the indicated time points (20 min, 1 hr, 6 hr, 12 hr and 24 hr). [Figure 4C] 1 is a histogram of the densitometric ratio of PTEN to β-actin showing increased PTEN levels. [Figure 5A]1 is a histogram showing the effect of BCN057 and 5-FU on the survival of pancreatic cancer cell lines after 72 hours of incubation of all drugs. Controls (ND) received no drug. Test groups included 5FU (50 μM), 057 (BCN057: 2, 5, 10, and 20 μM or 5FU+BCN057 (BCN057 at 2, 5, 10, and 20 μM in combination with 5FU at 50 μM). [Figure 5B] 13 is a histogram showing relative LGR-5 and BMI-1 expression in Panc-1 cells treated with 10 μM 057. [Figure 5C] A panel of images showing that BCN057 protects GI epithelium and LGR5 stem cells (green) from 5FU cytotoxicity. [Figure 5D] FIG. 13 is a histogram showing quantification of the % of LGR5+ cells remaining in the 1 mM compartment at the crypt base showing prevention of 5-FU-induced apoptosis in LGR5+ stem cells in the GI. [Figure 6] 1 is a histogram showing the cytotoxicity (% viability) of oxaliplatin and irinotecan individually or in combination with 5FU and BCN057 against Panc-1 cells. [Figure 7A] Western blot showing PD-1 expression on human T lymphocytes co-cultured with Panc-1 cells without drug (vehicle) or with BCN057 for 1, 6 and 12 hours. [Figure 7B] 1 is a histogram (concentration vs luminescence) showing that inhibition of GSK3β induces lymphocyte survival upon exposure to radiation. [Figure 8] Western blot showing PDL-1 expression (red arrows) on Panc-1 cells co-cultured with human T lymphocytes without drug (vehicle) or with BCN057 for 2.5, 5, 7.5 and 10 hours. [Figure 9] 1 is a graph of viability versus concentration of BCN077. Studies were performed with the human KRAS G12D mutant colorectal cancer cell line LS174T46 established from Dukes' type B adenocarcinoma BCN077, which shows potent activity. [Figure 10A]The structure of BCN076 is shown. [Figure 10B] The structure of BCN036 is shown. [Figure 10C] The structure of BCN077 is shown. [Figure 10D] The structure of BCN078 is shown. [Figure 11] 1 is a graph showing inhibition of proliferation of KRAS mutant G12D aggressive pancreatic cancer cell lines by BCN057, 076, 077, 036 and 078 analogs. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0050] definition Reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described with respect to that embodiment is included in at least one embodiment of the disclosure. Use of the phrase "in one embodiment" or "in an embodiment" in various places in this specification does not necessarily mean that they all refer to the same embodiment or embodiment, or to mutually exclusive alternative or alternative embodiments of other embodiments. Furthermore, various features are described that may be exhibited by some embodiments but not by others. Similarly, various requirements are described that may be requirements for some embodiments but not other embodiments. The terms embodiment and aspect may be used interchangeably in certain instances.

[0051] The terms used herein generally have their usual meaning in the technical field within the context of this disclosure and in the specific context in which each term is used.Specific terms used in the description of this disclosure are discussed below or elsewhere in this specification to provide additional guidance to practitioners regarding the description of this disclosure.It will be understood that the same thing can be stated in more than one way.

[0052] Thus, alternative terms and synonyms may be used for any one or more terms discussed herein. There is no special significance to whether a term is detailed or discussed herein. Synonyms for certain terms are provided. The description of one or more synonyms does not exclude the use of other synonyms. The use of examples anywhere in this specification, including examples of any term discussed herein, is for illustrative purposes only and is not intended to further limit the scope and meaning of the disclosure or of any exemplified term. Similarly, this disclosure is not limited to the various embodiments presented herein.

[0053] Without intending to further limit the scope of the present disclosure, the instruments, apparatus, methods and their related results according to the embodiments of the present disclosure are given below. It should be noted that headings or subheadings may be used in the examples for the convenience of the reader, but do not limit the scope of the present disclosure. Unless otherwise specified, all technical and scientific terms used herein generally have the same meaning as those understood by those skilled in the art to which this disclosure belongs. In case of conflict, this document, including definitions, shall prevail.

[0054] Unless otherwise indicated, all numbers expressing properties, items, amounts, parameters, properties, periods, etc. used in the specification and claims should be understood in all instances as being modified by the word "about". As used herein, the term "about" means that the property, item, amount, parameter, property, period so qualified encompasses a range of plus or minus 10% higher or lower than the value of the stated property, item, amount, parameter, property, period. Thus, unless otherwise indicated, the numerical parameters set forth in the specification and the appended claims are approximations that may vary. For example, since mass spectrometers may vary slightly in determining the mass of a given analyte, the term "about" in the context of the mass of an ion or the mass / charge ratio of an ion refers to ±0.50 atomic mass units. To be conservative, and without any attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical designation should be interpreted at least in light of the number of reported significant digits and by applying ordinary rounding techniques.

[0055] The use of the term "may" or "can" in the context of an embodiment or an aspect of an embodiment also has the alternative meaning of "may not" or "cannot." Thus, where the specification discloses that an embodiment or an aspect of an embodiment may or may not be included as part of the subject matter of the present invention, negative limitations or exclusions are likewise expressly intended, meaning that the embodiment or aspect of the embodiment may or may not be included as part of the subject matter of the present invention. Similarly, the use of the term "optionally" in the context of an embodiment or an aspect of an embodiment means that such embodiment or aspect of the embodiment may or may not be included as part of the subject matter of the present invention. Whether such a negative limitation or exclusion applies will be based on whether the negative limitation or exclusion is recited in the claimed subject matter.

[0056] Although the numerical ranges and values ​​describing the broad scope of the present disclosure are approximations, the numerical ranges and values ​​described in the specific examples are reported as precisely as possible. However, any numerical range and value inherently contains certain errors necessarily resulting from the standard deviations found in their respective testing measurements. The recitation of a range of values ​​herein is merely intended to serve as a shorthand method of individually referring to each separate value falling within the range. Unless otherwise indicated herein, each separate value of a numerical range is incorporated into the specification as if it were individually set forth herein.

[0057] The terms "a," "an," and "the," as used in the context of describing this disclosure (particularly in the context of the claims that follow), should be construed to encompass both the singular and the plural, unless otherwise indicated or clearly contradicted by context. Furthermore, order labels for specified elements, such as "first," "second," "third," and the like, are used to distinguish between elements, unless otherwise specified, and do not indicate or imply a required or limited number of such elements, or a particular location or order of such elements. All methods described herein can be performed in any suitable order, unless otherwise indicated herein or clearly contradicted by context. The use of any specific examples or exemplary language (e.g., "such as") described herein is intended merely to facilitate understanding of the disclosure, and does not limit the scope of the disclosure as otherwise claimed. No term in the specification should be construed as indicating any non-claimed element essential to the practice of the disclosure.

[0058] When used in the claims, whether added at the time of filing or by amendment, the open-ended transitional term "comprising" (and its equivalent open-ended transitional phrases such as including, containing, and having) includes all expressly recited elements, limitations, steps, and / or features, either alone or in combination with unrecited subject matter, and while the recited elements, limitations, and / or features are essential, other unrecited elements, limitations, and / or features may be added and still form a construct within the scope of the claim. Certain embodiments disclosed herein may be further limited in the claims by use of the closed transitional phrase "consisting of" or "consisting essentially of" in place of or as an amendment to "comprising." When used in the claims, whether added at the time of filing or by amendment, the closed transitional phrase "consisting of" excludes any element, limitation, step, or feature not expressly recited in the claim. The closed transitional phrase "consisting essentially of" limits the elements, limitations, steps and / or features explicitly recited in the claim and any and all other elements, limitations, steps and / or features that do not materially affect the basic and novel characteristics of the claimed subject matter. Accordingly, the meaning of the open transitional phrase "comprising" is defined to include all specifically recited elements, limitations, steps and / or features as well as any optional, unspecified inclusions.The meaning of the closed transitional phrase "consisting of" is defined as including only those elements, limitations, steps and / or features expressly recited in the claim, while the meaning of the closed transitional phrase "consisting essentially of" is defined as including only those elements, limitations, steps and / or features specifically recited in the claim and those elements, limitations, steps and / or features that do not materially affect the basic and novel characteristics of the claimed subject matter. Thus, the open transitional phrase "comprising" (and its equivalent open transitional phrases) includes, within its meaning, the claimed subject matter specified by the closed transitional phrase "consisting of" or "consisting essentially of," in the limiting case. Such embodiments described or claimed herein using the phrase "comprising" are expressly or essentially expressly described, enabled and established herein as opposed to the phrases "consisting of" and "consisting essentially of."

[0059] Where applicable, the terms "about" or "generally" as used herein and in the appended claims mean a tolerance of ±20% unless otherwise indicated. Also, where applicable, the term "substantially" as used herein and in the appended claims means a tolerance of ±10% unless otherwise indicated. It is to be understood that not all uses of the above terms can be numerically quantified such that a reference range is applicable.

[0060] The term "active agent" or "active ingredient" refers to a substance, compound, or molecule that is biologically active or that induces a biological or physiological effect on a subject to which it is administered. In other words, an "active agent" or "active ingredient" means the component of a composition for which the effect of the composition is responsible, in whole or in part. An active agent may be the primary active agent, or in other words, the component of a composition that is responsible for all or in part of the effect of the composition. An active agent may be a secondary agent, or in other words, the component of a composition that is an additional part of the composition and / or responsible for another effect.

[0061] The term "cancer" includes, but is not limited to, adenoid cystic carcinoma, adrenal gland cancer, amyloidosis, anal cancer, ataxia telangiectasia, atypical molar syndrome, aberrant nevus syndrome, basal cell carcinoma, cholangiocarcinoma, Birt-Hogg-Dube syndrome, bladder cancer, bone cancer, brain tumor, breast cancer, male breast cancer, carcinoid tumor, cervical cancer, colorectal cancer, ductal carcinoma, endometrial cancer, esophageal cancer, gastric cancer, gastrointestinal stromal tumor, and urinary tract cancer. Tumor (GIST), HER2 positive breast cancer, Islet cell tumor, Juvenile polyposis syndrome, Kidney cancer, Throat cancer, Leukemia - acute lymphoblastic leukemia, Leukemia - acute lymphocytic (ALL), Leukemia - acute myeloid AML, Leukemia - adult, Leukemia - childhood, Leukemia - chronic lymphocytic - CLL, Leukemia - chronic myeloid - CML, Liver cancer, Lobular carcinoma, Lung cancer, Lung cancer - small cell, Lymphoma - Hodgkin, Lymphoma - non-Hodgkin, Malignant glioma, Melanoma, Meningeal The cancers may include one or more of: myeloma, multiple myeloma, myelodysplastic syndromes (MDS), nasopharyngeal cancer, neuroendocrine tumors, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, pancreatic endocrine tumors, parathyroid cancer, penile cancer, peritoneal cancer, Peutz-Jeghers syndrome, pituitary tumors, polycythemia vera, prostate cancer, renal cell carcinoma, retinoblastoma, salivary gland cancer, sarcoma, sarcoma-Kaposi, skin cancer, small intestine cancer, gastric cancer, testicular cancer, thymoma, thyroid cancer, uterine (endometrial) cancer, vaginal cancer, and Wilms' tumor. The term "blood cancer" may include one or more of leukemia, lymphoma, myeloma, myelodysplastic syndrome, and myeloproliferative disorder.

[0062] The term "derivative" may refer to any compound that has the same or similar core structure as the compound, but has at least one structural difference, including substitution, deletion, and / or addition of one or more atoms or functional groups. The term "derivative" does not imply that the derivative is synthesized from the parent compound as a starting material or intermediate, even if this is the case. The term "derivative" may include prodrugs, or metabolites of the parent compound. Derivatives include compounds in which carboxyl groups in the parent compound are derivatized to form methyl and ethyl esters, or other types of esters or hydrazides. Derivatives include compounds in which hydroxyl groups in the parent compound are derivatized to form O-acyl or O-alkyl derivatives. Derivatives include compounds in which a hydrogen bond donor group in the parent compound is replaced with another hydrogen bond donor group, such as OH, NH, or SH. Derivatives include replacing a hydrogen bond acceptor group in the parent compound with another hydrogen bond acceptor group, such as an ester, ketone, carbonate, tertiary amine, imine, thione, sulfone, tertiary amide, and sulfide. Derivatives can also include salt forms, such as pharma- ceutically acceptable salt forms of a parent compound or derivatives thereof.

[0063] The term "fibrosis" refers to a condition involving the formation of excess fibrous connective tissue in an organ or tissue during repair or reaction. This can be a reactive, benign, or pathological condition.

[0064] As used herein, "mitigating" means reducing one or more negative symptoms of a condition compared to a cell, organ, tissue, or organism exhibiting the symptom or condition that is not treated, but for the same amount of time.

[0065] In some embodiments, contacting a cell, organ, tissue, or organism with a compound of the invention can include administering to the subject a therapeutically effective amount of the compound. As used herein, a "therapeutically effective amount" is an amount sufficient to alleviate a negative symptom or condition.

[0066] The term "subject" or "patient" refers to any single animal, more preferably a mammal, for which treatment is desired (including, for example, dogs, cats, horses, rabbits, zoo animals, cows, pigs, sheep, and non-human animals such as non-human primates). Most preferably, the patient herein is a human.

[0067] As used herein, the term "pharmaceutically acceptable carrier" refers to any and all solvents, dispersion media, coatings, isotonic and absorption delaying agents, etc., that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. The composition may also contain other active compounds that provide supplementary, additional, or enhanced therapeutic effects.

[0068] As used herein, the term "pharmaceutically acceptable composition" refers to a composition comprising at least one compound disclosed herein formulated together with one or more pharma- ceutically acceptable carriers.

[0069] As used herein, the term "pharmaceutical acceptable prodrug" refers to prodrugs of compounds of the present disclosure, and where possible, zwitterionic forms of compounds of the present disclosure, that are suitable for use in contact with the tissues of humans and lower animals without excessive toxicity, irritation, and allergic response, within the scope of sound medical judgment, and are effective for their intended use, commensurate with a reasonable benefit / risk ratio. Discussion is provided in Higuchi et al., "Prodrugs as Novel Delivery Systems," ACS Symposium Series, Vol. 14, and Roche, EB, ed. Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987. Both of these documents are incorporated herein by reference.

[0070] The term "pharmaceutically acceptable salts" refers to salts of acidic or basic groups that may be present in the compounds used in the compositions. Compounds contained in the compositions that are basic in nature can form a wide variety of salts with various inorganic and organic acids. Acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are those that form non-toxic acid addition salts, i.e., salts with pharmaceutically acceptable anions, including, but not limited to, sulfate, citrate, malate, acetate, oxalate, chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, oleate, These include tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)). Compounds contained in the present composition that contain an amino moiety can form pharma- ceutically acceptable salts with various amino acids in addition to the acids mentioned above. Compounds contained in the present composition that are acidic in nature can form base salts with various pharma- ceutically acceptable cations. Examples of such salts include alkali metal or alkaline earth metal salts and, in particular, calcium, magnesium, sodium, lithium, zinc, potassium, and iron salts.

[0071] The term "chemotherapy" refers to a type of cancer treatment that uses one or more anti-cancer drugs (i.e., chemotherapeutic agents). Chemotherapy may be given for curative purposes (usually a combination of drugs), or it can be used to extend life or relieve symptoms (i.e., palliative chemotherapy). Traditional chemotherapy agents are cytotoxic by interfering with cell division (mitosis). Common side effects of chemotherapy include bone marrow suppression, mucositis (inflammation of the lining of the digestive tract), and alopecia.

[0072] The term "KRAS" refers to a gene that acts as an "on / off" switch in intracellular signaling. When it functions normally, it controls cell proliferation. It is allosterically activated and, together with other intracellular signaling receptors such as c-Raf and PI3-kinase, recruits and activates proteins required for growth factors proliferation. When it is mutated, the negative signaling is disrupted, causing cells to hyperproliferate and develop into cancer.

[0073] The term "PD-1", "programmed cell death protein 1" or "CD279" refers to a protein on the surface of T and B cells that has a role in regulating the immune system's response to cells in the human body by downregulating the immune system by suppressing T cell inflammatory activity and promoting self-tolerance. This prevents autoimmune disease, but it can also interfere with the immune system killing cancer cells. PD-1 is an immune checkpoint that protects against autoimmunity by two mechanisms. First, it promotes apoptosis (programmed cell death) of antigen-specific T cells in lymph nodes. Second, it reduces apoptosis of regulatory T cells (anti-inflammatory, suppressive T cells). PD-1 inhibitors, a class of drugs that block PD-1, are used to activate the immune system to attack tumors and treat certain types of cancer. Binding of PD-1 by either of its ligands, PD-L1 or PD-L2, on neighboring cells suppresses TCR signaling and TCR-mediated proliferation, transcriptional activation and cytokine production. This prevents autoimmune disease, but it can also prevent the immune system from killing cancer cells. Therapeutic antibodies designed to block the PD-1 / PD-L1 interaction have potential for the treatment of cancer.

[0074] Programmed death-ligand 1 (PD-L1), also known as cluster of differentiation 274 (CD274) or B7 homolog 1 (B7-H1), is a protein that in humans is encoded by the CD274 gene. PD-L1 binds to its receptor, PD-1, which is found on activated T cells, B cells, and myeloid cells, and regulates activation or inhibition.

[0075] "Immunomodulatory agent" refers to a substance, agent, signaling pathway, or component thereof that modulates the immune response. "Regulating," "modifying," or "modulating" the immune response refers to any change in the cells of the immune system or in the activity of such cells. Such modulation includes stimulation or suppression of the immune system, which may be manifested by an increase or decrease in the number of various cell types, an increase or decrease in the activity of these cells, or any other change that may occur within the immune system. Both inhibitory and stimulatory immunomodulatory agents have been identified, some of which may have enhanced function in cancer, infectious diseases, or neurodegenerative microenvironments.

[0076] The term "immunotherapy" refers to the treatment of a subject suffering from a disease or at risk of recurrence of a disease by methods that involve inducing, enhancing, suppressing or modifying the immune response. "Treatment" or "therapy" of a subject refers to any type of intervention or procedure performed on a subject, or the administration of an active agent to a subject, for the purpose of ameliorating, alleviating, ameliorating, inhibiting, delaying or preventing the onset, progression, development, severity or recurrence of symptoms, complications, conditions, or biochemical indicators associated with a disease.

[0077] The term "glycogen synthase kinase 3β" or "GSK3β" refers to an enzyme that in humans is encoded by the GSK3B gene. It is integrally involved in pathways of cell proliferation and apoptosis. GSK-3β has been shown to phosphorylate β-catenin, thus targeting it for degradation. GSK-3β is thus part of the canonical β-catenin / Wnt pathway, which signals cells to divide and proliferate. It also participates in multiple apoptotic signaling pathways by phosphorylating transcription factors that regulate apoptosis. GSK-3β is also overexpressed in several types of cancer, such as colorectal, ovarian, and prostate cancer. GSK-3β inhibitors are also useful in the treatment of Alzheimer's disease, stroke, and mood disorders, including bipolar disorder. In diabetes, GSK-3β inhibitors increase insulin sensitivity, glycogen synthesis, and glucose metabolism in skeletal muscle, and reduce obesity by acting on the adipogenic process.

[0078] The term "SFRP" or "secreted frizzled-related protein" refers to Wnt signaling pathway inhibitors that are part of the soluble frizzled-related proteins (sFRPS). sFRPS function as regulators of Wnt signaling through direct interaction with Wnt. Five mammalian sFRPSs have been identified (sFRP-1, sFRP-2, sFRP-3, sFRP-4 and sFRP-5). These proteins consist of approximately 300 amino acids, including a signal sequence, a frizzled-like cysteine-rich domain (CRD), and a small hydrophilic C-terminal domain. As a group, sFRPSs are expressed in a variety of embryonic and adult tissues, suggesting a common mechanism for inhibiting Wnt signaling. However, individual family members have specific spatial and temporal expression patterns.

[0079] The term "Wnt signaling pathway" refers to a group of signaling pathways that pass signals to cells through cell surface receptors. The canonical Wnt pathway results in the regulation of gene transcription.

[0080] The term "RIGS" or "Radiation-Induced Gastrointestinal Syndrome" arises from a combination of the direct cytocidal effects of radiation on intestinal crypt and stromal cells, with symptoms ranging from loss of mucosal barrier and diarrhea to electrolyte imbalance, weight loss and death. A significant proportion of patients experience radiation-induced toxicity due to damage to normal tissues in the radiation field. The use of chemical or biological approaches aimed at reducing or preventing radiation therapy-induced normal tissue toxicity has been a long-standing goal.

[0081] The term "radiation therapy" or "radiotherapy" refers to therapy that uses ionizing radiation, usually as part of cancer treatment to control or kill malignant cells. Radiation therapy is usually applied to cancerous tumors for its ability to control cell growth. Ionizing radiation works by damaging the DNA of cancerous tissue, resulting in cell death. The main side effects are fatigue and skin irritation. Acute side effects can include nausea and vomiting, damage to epithelial surfaces, mouth, pharynx, and stomach upset, swelling, intestinal discomfort. RIGS (Radiation Induced Gastrointestinal Syndrome) is a general term used to describe side effects related to the gastrointestinal system.

[0082] The term "treating" or "treatment" refers to one or more of: (1) inhibiting a disease; for example, inhibiting a disease, condition, or disorder in an individual experiencing or exhibiting the pathology or symptomology of the disease, condition, or disorder (i.e., preventing further progression of the pathology and / or symptomology); and (2) alleviating a disease; for example, alleviating a disease, condition, or disorder in an individual experiencing or exhibiting the pathology or symptomology of the disease, condition, or disorder (i.e., ameliorating the pathology and / or symptomology), for example, reducing the severity of the disease. Treatment can be prophylactic and / or therapeutic. The term "prophylactic or therapeutic" treatment is art-recognized and includes administration of one or more compositions of interest to the host. If it is administered prior to clinical symptoms of an undesirable condition (e.g., a disease or other undesirable condition in a host animal), the treatment is prophylactic (i.e., it protects the host against the onset of the undesirable condition), whereas if it is administered after manifestation of the undesirable condition, the treatment is therapeutic (i.e., intended to attenuate, ameliorate, or stabilize an existing undesirable condition or its side effects).

[0083] As used herein, the term "unit dosage form" or "unit" refers to a physically discrete unit suitable as a unitary dosage for human and animal subjects, each unit containing a predetermined amount of compound calculated to be sufficient to produce a desired effect together with a pharma- ceutically acceptable diluent, carrier or vehicle. The specifications for the novel unit dosage forms of the present disclosure depend on the particular compound used and the effect to be achieved, and the pharmacodynamics associated with each compound in the subject.

[0084] The compounds of the present disclosure may contain one or more chiral centers and / or double bonds and therefore exist as stereoisomers, such as geometric isomers, enantiomers or diastereomers. The term "stereoisomers" as used herein consists of all geometric isomers, enantiomers or diastereomers. These compounds may be named by the abbreviations "R" or "S" depending on the arrangement of the substituents around the stereogenic carbon atom. The present disclosure encompasses various stereoisomers of these compounds and mixtures thereof. Stereoisomers include enantiomers and diastereomers. A mixture of enantiomers or diastereomers may be named in the nomenclature "(±)", but one of skill in the art will understand that the structure may implicitly represent a chiral center.

[0085] Individual stereoisomers of the compounds of the present disclosure can be prepared synthetically from commercially available starting materials containing asymmetric or chiral centers, or by preparation of racemic mixtures followed by separation methods well known to those skilled in the art. These separation methods are typified by (1) binding of the enantiomeric mixture to a chiral auxiliary, recrystallization or chromatographic separation of the resulting diastereomeric mixture and liberation of the optically pure product from the auxiliary, (2) salt formation using an optically active resolving agent, or (3) direct separation of the mixture of optical enantiomers on a chiral chromatographic column. Stereoisomeric mixtures can also be resolved into their component stereoisomers by well-known methods such as chiral phase gas chromatography, chiral phase high performance liquid chromatography, crystallization of the compound as a chiral salt complex, or crystallization of the compound in a chiral solvent. Stereoisomers can also be obtained from stereomerically pure intermediates, reagents, and catalysts by well-known asymmetric synthetic methods.

[0086] Geometric isomers may also exist in the compounds of the present disclosure. The present disclosure encompasses various geometric isomers and mixtures thereof resulting from the arrangement of substituents around a carbon-carbon double bond or the arrangement of substituents around a carbon ring. Substituents around a carbon-carbon double bond are designated as having the "Z" or "E" configuration, with "Z" and "E" being used according to IUPAC standards. Unless otherwise specified, structures depicting double bonds encompass both E and Z isomers.

[0087] Substituents around a carbon-carbon double bond may alternatively be referred to as "cis" or "trans", with "cis" referring to substituents on the same side of the double bond and "trans" referring to substituents on opposite sides of the double bond. The arrangement of substituents around a carbon ring is designated as "cis" or "trans". The term "cis" refers to substituents on the same side of the plane of the ring and "trans" refers to substituents on opposite sides of the plane of the ring. A mixture of compounds in which substituents are located on both the same and opposite sides of the plane of the ring is referred to as "cis / trans".

[0088] The compounds disclosed herein may exist as tautomers, and both tautomeric forms are intended to be encompassed within the scope of the disclosure even if only one tautomeric structure is shown.

[0089] The term "alkoxy" refers to a chemical substituent of formula -OR, where R is an optionally substituted C1-C6 alkyl group, unless otherwise specified. In some embodiments, the alkyl group can be substituted, for example, an alkoxy group can have 1, 2, 3, 4, 5, or 6 substituents, as defined herein.

[0090] The term "alkoxyalkyl" refers to a heteroalkyl group, as defined herein, and is considered an alkyl group substituted with an alkoxy group. Exemplary unsubstituted alkoxyalkyl groups contain 2-12 carbons. In some embodiments, alkyl and alkoxy can be further substituted with 1, 2, 3, or 4 substituents, as defined herein for each group.

[0091] The terms "alkyl", "alkenyl" and "alkynyl" include straight chain, branched chain and cyclic monovalent substituents, and combinations thereof, which, if unsubstituted, contain only C and H. Examples include methyl, ethyl, isobutyl, cyclohexyl, cyclopentylethyl, 2-propenyl, 3-butynyl, and the like. As used herein, the term "cycloalkyl", unless otherwise specified, represents a monovalent saturated or unsaturated non-aromatic cyclic alkyl group having 3 to 9 carbons (e.g., C3-C9 cycloalkyl), and is exemplified by cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclo[2.2.1.]heptyl, and the like. When a cycloalkyl group contains one carbon-carbon double bond, the cycloalkyl group may be referred to as a "cycloalkenyl" group. Exemplary cycloalkenyl groups include cyclopentenyl, cyclohexenyl, and the like. Typically, alkyl, alkenyl and alkynyl groups are alkyl groups having 1 to 12 carbons (e.g., C1-C 12 alkyl) or 2-12 carbons (e.g., C2-C 12 Alkenyl or C2-C 12 In some embodiments, the alkyl group is C1-C8, C l or a C2-C8, C2-C6, C2-C4, or C2-C3 alkenyl or alkynyl group. Additionally, any hydrogen atom on one of these groups can be substituted with a substituent described herein.

[0092] Heteroalkyl, heteroalkenyl and heteroalkynyl are similarly defined and contain at least one carbon atom, but also contain one or more O, S or N heteroatoms or combinations thereof in the backbone residue, whereby each heteroatom in a heteroalkyl, heteroalkenyl or heteroalkynyl replaces one carbon atom of the corresponding alkyl, alkenyl or alkynyl group of the heterotype. In some embodiments, heteroalkyl, heteroalkenyl and heteroalkynyl groups have a C at each end where the group is attached to the other group, and the heteroatoms present are not located at the terminal positions. As understood in the art, these heterotypes do not contain more than three adjacent heteroatoms. In some embodiments, the heteroatom is O or N. As used herein, the term "heterocyclyl" refers to, for example, a cyclic heteroalkyl or heteroalkenyl that is a 3-, 4-, 5-, 6- or 7-membered ring containing 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur, unless otherwise specified. The 5-membered ring has 0-2 double bonds, and the 6- and 7-membered rings have 0-3 double bonds. The term "heterocyclyl" also refers to heterocyclic compounds having bridged polycyclic structures in which one or more carbon and / or heteroatoms bridge two non-adjacent members of a monocyclic ring, such as quinuclidinyl groups. The term "heterocyclyl" includes bicyclic, tricyclic, and tetracyclic groups in which any of the above heterocyclic rings are fused to one, two, or three carbon rings, such as an aryl ring, a cyclohexane ring, a cyclohexene ring, a cyclopentane ring, a cyclopentene ring, or another monocyclic heterocyclic ring, such as indolyl, quinolyl, isoquinolyl, tetrahydroquinolyl, benzothienyl groups, and the like.

[0093] As used herein, the term "alkylsulfonyl" refers to a heteroalkyl group, as described herein, described as an optionally substituted alkyl group, including an -S(O)2- group.

[0094] As used herein, the term "amino" refers to -N(R), where each R is independently H, OH, NO, N(R), SOOR, SOR, SOR, SON(R), an N-protecting group, alkyl, alkenyl, alkoxy, aryl, alkaryl, cycloalkyl, alkcycloalkyl, heterocyclyl (e.g., heteroaryl), alkheterocyclyl (e.g., alkheteroaryl), or two R combined to form a heterocyclyl or an N-protecting group and each R N2 is independently H, alkyl, or aryl. In a preferred embodiment, amino is -NH2 or -NHR, and R is independently OH, NO2, NH2, NR2, SO2OR, SO2R, SOR, SON(R)2, SON(R), alkyl, or aryl, and each R can be H, alkyl, or aryl. As used herein, the term "aminoalkyl" refers to a heteroalkyl group, as defined herein, described as an alkyl group substituted with an amino group, as defined herein. Each of the alkyl and amino can be further substituted with 1, 2, 3, or 4 substituents, as defined herein for each group. For example, the alkyl moiety can include an oxo (=O) substituent.

[0095] An "aromatic" or "aryl" moiety refers to any monocyclic or fused bicyclic ring system that has the characteristics of aromaticity in terms of electron distribution throughout the ring system, including monocyclic or fused bicyclic moieties such as phenyl or naphthyl; "heteroaromatic" or "heteroaryl" also refers to such monocyclic or fused bicyclic ring systems that contain one or more heteroatoms selected from O, S, and N. The inclusion of heteroatoms allows for the inclusion of 6-membered rings as well as 5-membered rings that are considered aromatic. Thus, typical aromatic / heteroaromatic systems include pyridyl, pyrimidinyl, indolyl, benzimidazolyl, benzotriazolyl, isoquinolyl, quinolyl, benzothiazolyl, benzofuranyl, thienyl, furyl, pyrrolyl, thiazolyl, oxazolyl, isoxazolyl, benzoxazolyl, benzisoxazolyl, imidazolyl. Phthalimide is also considered aromatic, since tautomers are theoretically possible. Typically, the ring system contains 5 to 12 ring atoms or 6 to 10 ring atoms. In some embodiments, the aromatic or heteroaromatic moiety is a 6-membered aromatic ring system that may optionally contain 1 to 2 nitrogen atoms. More particularly, the moiety is an optionally substituted phenyl, pyridyl, indolyl, pyrimidinyl, pyridazinyl, benzothiazolyl or benzimidazolyl, pyrazolyl, imidazolyl, isoxazolyl, thiazolyl, benzothiazolyl, indolyl. More particularly, such a moiety is phenyl, pyridyl, or pyrimidinyl, and even more particularly, it is phenyl. "O-aryl" or "O-heteroaryl" refers to an aromatic or heteroaromatic system that is linked to another residue via an oxygen atom. A typical example of an O-aryl is phenoxy. Similarly, "arylalkyl" refers to an aromatic and heteroaromatic system, including heteroforms thereof, that is bonded to another residue via a carbon chain that is saturated or unsaturated, typically C1-C2, C1-C6, or, more specifically, C1-C4 or C1-C3 if saturated, or C2-C8, C2-C6, C2-C4, or C2-C3 if unsaturated.For clarity, arylalkyl thus includes an aryl or heteroaryl group as defined above linked to an alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl or heteroalkynyl moiety as also defined above. Exemplary arylalkyls include aryl (C6-C. 12 ) Alkyl (C1-C8), Aryl (C6-C 12 ) alkenyl (C2-C8), or aryl (C6-C 12 ) alkynyl(C2-C8), as well as their hetero forms. A typical example is phenylmethyl, commonly called benzyl.

[0096] Halo may be any halogen atom, in particular F, Cl, Br, or I, and more particularly fluoro or chloro.

[0097] As used herein, the term "haloalkyl" refers to an alkyl group, as defined herein, substituted with a halogen group (i.e., F, Cl, Br, or I). Haloalkyl can be substituted with one, two, three, or, in the case of alkyl groups of two or more carbons, four halogens. Haloalkyl groups include perfluoroalkyl. In some embodiments, haloalkyl groups can be further substituted with one, two, three, or four substituents as described herein for alkyl groups.

[0098] As used herein, the term "hydroxyl" refers to an --OH group.

[0099] As used herein, the term "hydroxyalkyl" refers to an alkyl group, as defined herein, substituted with one to three hydroxy groups, provided that only one hydroxy group may be attached to a single carbon atom of the alkyl group, and is exemplified by hydroxymethyl, dihydroxypropyl, and the like.

[0100] In general, a substituent (e.g., alkyl, alkenyl, alkynyl, or aryl (including heterotypes as defined above) may itself be optionally substituted with additional substituents. The nature of these substituents is similar to that detailed with respect to substitution on the base structure above. Thus, if a substituent embodiment is an alkyl, the alkyl may be optionally substituted with the remaining substituents listed as substituents, provided that this is chemically reasonable and that this does not violate the size limit of the alkyl itself; for example, substitution of an alkyl by an alkyl or by an alkenyl merely extends the upper carbon atom limit for these embodiments and is not incorporated. For example, if a group is substituted, the group may be substituted with 1, 2, 3, 4, 5, or 6 substituents. Optional substituents include, but are not limited to, C1-C6 alkyl or Heteroaryl, C2-C6 alkenyl or heteroalkenyl, C2-C6 alkynyl or heteroalkynyl, halogen; aryl, heteroaryl, azido (-N3), nitro (-NO2), cyano (-CN), acyloxy (-OC(=O)R'), acyl (-C(=O)R'), alkoxy (-OR'), amido (-NR'C(=O)R" or -C(=O)NR'R''), amino (-N R'R''), carboxylic acid (-COH), carboxylate ester (-COR'), carbamoyl (-OC(=O)NR'R" or -NRC(=O)OR'), hydroxy (-OH), isocyano (-NC), sulfonate (-S(=OHOR), sulfonamide (-S(=OHNRR' or -NRS(=O)2R'), or sulfonyl (-S(=O)2R), where each R or R' is independently selected from H, C1-C6 alkyl or heteroaryl, C2-C6 alkenyl or heteroalkenyl, C2-C6 alkynyl or heteroalkynyl, aryl, or heteroaryl. The substituents may have, for example, 1, 2, 3, 4, 5, 6, 7, 8, or 9 substituents.

[0101] Exemplary optional substituents are independently halo, CN, NO2, CF3, OCF3, COOR, CONR Y2, OR, SR, SOR, SO2R, NR2, NR(CO)R, NRC(O)OR, NRC(O)NR2, NRSO2NR2, or NRS02R, where each R is independently H or an optionally substituted group selected from alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl, and aryl (all as defined above); or the substituents can be optionally substituted groups selected from alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, aryl, heteroaryl, O-aryl, O-heteroaryl, and arylalkyl.

[0102] Other technical terms used herein have their ordinary meaning in the art in which they are used as exemplified by various technical dictionaries. The specific values ​​and configurations discussed in these non-limiting examples may vary, and they are mentioned merely to illustrate at least one embodiment and are not intended to limit their scope.

[0103] Detailed Description of the Invention It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the present technology as claimed. Additional features and advantages of the present technology will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by the practice of the present technology. The advantages of the present technology will be realized and attained by the structure particularly pointed out in the description and claims hereof.

[0104] BCN057 and Analogs BCN057, also known as YEL002, is represented by the following structure: [ka] It is understood that the present invention encompasses the compound BCN057, its analogs and salts. In one embodiment, the present invention relates to the compound BCN057 in the form of a free base. In another embodiment, the present invention relates to the compound BCN057 or a pharma- ceutically acceptable salt thereof.

[0105] Due to their availability in medicines, salts of the compounds of BCN057 may be preferred as pharma- ceutically acceptable salts. Suitable pharma-ceutically acceptable salts may include acid addition salts. The pharma-ceutically acceptable salts may be easily prepared by using a desired acid or base as needed. The resulting salts may be precipitated from the solution and collected by filtration or may be recovered by evaporation of the solvent. The compounds may exist as stereoisomers, tautomers, pharma-ceutically acceptable salts, or hydrates thereof. Some analogs of BCN057 are shown below.

[0106] An embodiment is a compound of formula B: [ka] and a method of treatment comprising administering a therapeutically effective amount of a compound having the structure R 1 is an alkyl, alkylamine, or ether; R 2 is H or CH3; and R 3 is H or OH.

[0107] In some embodiments, the compound is an analog selected from Formulas II-XIX, including Compound H, Compound K, Compound BNB-1, Compound BNB-2, Compound BNB-3, Compound BNB-4, and Compound BNB-5, as shown in Table 2 below.

[0108] One embodiment is a compound of formula I, or an analog thereof, as detailed below. [ka]

[0109] BCN057, together with the compounds and analogs disclosed herein, can be prepared according to art-established methods of organic synthesis.General methods of compound synthesis can be found, for example, in Stuart Warren and Paul Wyatt, Workbook for Organic Synthesis: The Disconnection Approach, second Edition, Wiley, 2010.Exemplary methods of compound manufacture are provided in US Patent Application No. 13 / 813,923 and US Patent Application No. 14 / 889,719.These patent applications are incorporated herein by reference.The compound also includes the pharma- ceutically acceptable salts of the compound, its prodrugs, its hydrates, its solvates and its polymorphic crystals.The compound can be administered as a pharmaceutical composition.

[0110] Usefulness and Administration BCN057 and BCN077 and analogs described herein are useful for use in the methods of the present invention, and, without being bound by theory, are believed to exert their desired effects in part through their ability to regulate Wnt-β-catenin signaling. The Wnt pathway is involved in tissue development in embryos and tissue maintenance in adults. It controls a series of specific genes that control cell growth, migration and cell survival. Prolonged activation of these genes and abnormal activation of the Wnt pathway can lead to uncontrolled cell growth and survival, which can promote cancer formation in a series of tissues, including colon, liver and ovary.

[0111] The Wnt signaling pathway is a group of signal transduction pathways initiated by proteins that pass signals into cells through cell surface receptors. Aberrant activation of the Wnt pathway has been linked to human cancers, particularly cancers of the gastrointestinal (GI) tract. Inhibition of aberrant Wnt pathway activity in cancer cell lines can block their proliferation, offering the potential for novel therapeutic agents.

[0112] Wnt ligands bind to LRP5 / 6 and Frizzled co-receptors present on epithelial crypt cells, resulting in β-catenin stabilization and nuclear translocation, where it binds to the nuclear transcription factor TCF4 to promote gene expression programs that support stem cell maintenance, proliferation and differentiation. Activation of Wnt / β-catenin signaling is also important for crypt regeneration after injury. Several studies have demonstrated that Respondin 1 (RSPO1), an ISC growth factor and LGR5 receptor agonist, activates the Wnt / β-catenin pathway to repair and regenerate the intestine after chemotherapy-radiation-induced injury. DKK1, a negative regulator of the Wnt / β-catenin pathway, impairs RSPO1-induced intestinal regeneration.

[0113] In the absence of Wnt signals, the transcriptional activator β-catenin is actively degraded in cells. Phosphorylated β-catenin is then recognized and ubiquitinylated, leading to its proteasomal degradation. The levels of free β-catenin consequently remain low, which allows DNA-binding T-cell factor / lymphoid factor enhancer factor (Tcf / Lef) proteins to interact with transcriptional co-repressors to block target gene expression in the nucleus.

[0114] In addition to treating cancer, therapeutic agents that regulate Wnt / β-catenin signaling also have potential in treating other diseases, including fibrosis, inflammatory conditions, bone growth, wound healing, osteoporosis, alopecia (i.e., hair loss), depression, and viral infections. Thus, there is a need for compounds and methods that can inhibit and / or modulate Wnt / β-catenin signaling.

[0115] BCN057 and Analogs - Cancer Treatments and Therapies Several studies have demonstrated that the Wnt pathway is persistently activated in both hereditary familial adenomatous polyposis and spontaneous forms of colon cancer. Prolonged activation of the Wnt pathway in these cells promotes their proliferation into benign adenomas (also called polyps), which often progress to invasive colon carcinomas. Approximately 90% of sporadic colon cancers exhibit aberrant Wnt signaling activity, usually as a result of mutations in APC.

[0116] BCN057 and BCN077 are antineoplastic small molecules that are effective in treating cancer, especially in treating pancreatic and gastrointestinal (GI) cancer. It functions by dual action. It induces apoptosis in cancer cells. Secondly, it can promote epithelial cell proliferation and survival. BCN057 and BCN077 are particularly effective against pancreatic and gastrointestinal (GI) cancer. BCN057 and BCN077 have shown antineoplastic effects against "KRAS" cancer.

[0117] Combined use with radiation / chemotherapy BCN057 and BCN077 are effective in treating cancer, particularly in treating pancreatic and gastrointestinal (GI) cancer. In one aspect, disclosed herein is a method of treating radiation-induced gastrointestinal syndrome (RIGS) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of BCN057 and BCN077, or an analog thereof. The analog may be one or more of the compounds of formulas II-XIX. BCN057 (and BCN077) reduces RIGS and improves the therapeutic ratio of abdominal radiotherapy. In addition, BCN057 and BCN077 reduces radiation-induced mucositis, including oral mucositis, e.g., pharyngeal, gastric and intestinal GI mucositis, enteritis and proctitis. In addition, it can prevent and treat damage to epithelial tissues from radiation and chemotherapy. The compounds are also useful for treating or preventing these radiation syndromes associated with radiation therapy.

[0118] Combination therapy Combination therapy may be particularly effective with drugs that work by different mechanisms, thereby reducing the possibility of developing resistant cancer cells. When drugs with different effects are combined, each drug can be used at its optimal dose without intolerable side effects. BCN057 (and BCN077) and a second drug can be combined for therapeutic effect. The second drug can have a different mechanism. Alternatively, it can use the same mechanism as BCN057 for therapeutic effect. The combination can work through a synergistic effect. Combination therapy can also include additional (e.g., third, fourth, fifth, etc.) drugs. BCN512 can be used in combination therapy in the same manner.

[0119] Treatment of viral infections Additional embodiments include the use of BCN057, BCN077 and their analogs for the treatment of viral infections. Viral disease infections occur when the body is invaded by a pathogenic virus and infectious viral particles attach to and invade susceptible cells. The host immune response can mediate disease and hyperinflammation. Stimulation of the innate and adaptive immune system in response to viral infections destroys infected cells, which can lead to severe pathological consequences for the host (i.e., virus-induced immunopathology). In particular, immunopathology results from the excessive release of antibodies, interferons and pro-inflammatory cytokines, activation of the complement system, or hypermobility of cytotoxic T cells. Secretion of interferons and other cytokines can induce cell damage, fever, and flu-like symptoms. In severe cases of certain viral infections, such as in the 2005 avian H5N1 influenza, aberrant induction of the host immune response can induce a flaring release of cytokines, known as a cytokine storm.

[0120] Wnt signaling is important for the innate immune response to viruses. Pathogenic viruses suppress the expression of critical genes downstream of β-catenin to evade the first line of defense of the immune system. β-catenin is essential for the expression of IFNα / β and the subsequent transcriptional activation of interferon-inducible genes. IFNβ induces auto- and paracrine expression of antiviral genes. Thus, BCN057, BCN077 and analogs prevent pathological outcomes such as virus-induced immunopathology.

[0121] Treating depression Additional embodiments include the use of BCN057, BCN077 and analogs for the treatment of depression.Wnt signaling is also important for treating depression.Since the 1950s, lithium carbonate and other salts have been used to treat depression as well.Lithium functions by inhibiting GSK3B in the wnt downstream signaling pathway and transcribing the gene downstream of β-catenin.

[0122] Recent studies have shown that altered Wnt signaling may be involved in the pathophysiology of mood disorders. β-catenin levels are reduced in the hippocampal CA3 and CA4 regions, and Wnt levels are increased in the hippocampal CA4 region of postmortem schizophrenic brains. Lithium inhibits the canonical Wnt pathway component, GSK3β, and this pathway has been proposed as a specific target in the treatment of bipolar disorder. Therefore, BCN057, BCN077 and analogs can be used to treat depression and mood disorders.

[0123] PD-1 Modulation and Treatment of Diseases Associated with PD-1 Activity PD-1 is the receptor for PD-L1 and PD-L2. The PD-1 / PD-L signaling pathway plays a central role in peripheral immune tolerance, which prevents inappropriate immune responses under physiological conditions. PD-1 is an immune checkpoint inhibitor shown to reduce immune system responses in cancer and chronic infections. However, in infections or cancer, another role of this pathway leading to immune suppression may cause significant problems.

[0124] Several studies have shown that PD-1 is involved in various diseases, including cancer, autoimmune diseases, chronic infections and sepsis. Such studies highlight the dual role of PD-1 in immune tolerance, and reduction of PD-1 leads to autoimmune diseases. Depletion of PD-1+ T cells has beneficial effects in autoimmune diseases, slowing inflammation and disease progression. Reduction of PD-1 predisposes to autoimmune diseases, as described in experiments of PD-1 blockade or knockout in mice. PD-1 may also be a target for immunotherapy in multiple sclerosis (MS).

[0125] The role of PD-1 in cancer has been extensively studied. The expression of PD-L1 and / or PD-L2, either in tumors or in infiltrating immune cells, has been validated in numerous tumors, indicating the role of the PD-1 / PD-L1 axis as a prognostic trait and therapeutic target. Autoimmune diseases are defined as an abnormal immune response against an organism's own cells and tissues. The incidence of autoimmune diseases is increasing worldwide, and PD-1 and PD-L have been shown to be involved in regulating both central and peripheral immune tolerance. In particular, during thymocyte development, PD-1 plays a key role by regulating the signaling threshold during positive selection. Thus, the population of CD4+CD8+ thymocytes expands in the absence of PD-1 or PD-L1. Furthermore, the PD-1 pathway acts as a negative regulator in autoreactive T and B cells to maintain immune tolerance.

[0126] BCN057, BCN077 and analogs described herein are useful in the methods of the present invention, and without being bound by theory, are believed to exert their effects in part due to their ability to modulate PD-1 activity. In particular, applicants propose the use of therapeutic BCN057 and analogs of BCN057 to modulate PD-1 activity. Thus, BCN057 can treat certain diseases in which PD-1 is involved. The mechanism involves GSK3β inhibition.

[0127] In particular, applicants propose that BCN057 and BCN077 inhibit GSK3β, thereby inducing a proapoptotic phosphorylation pattern on c-Jun in KRAS G12D mutant pancreatic cancer cells, leading to restoration of PTEN expression and consequent apoptosis. Concurrent with their GSK3β inhibition, BCN057 (and BCN077) are small molecule inhibitors of PD-1 expression on human T lymphocytes co-cultured with human pancreatic cancer cells. BCN057 and BCN077 can promote synthetic lethality specifically against malignant cells, and thus should be considered to improve the therapeutic ratio in pancreatic and epithelial cancer treatment in conjunction with chemotherapy and radiotherapy.

[0128] Chronic infections are characterized by persistent high-level antigen exposure, causing T cells to gradually lose effector function and progress to exhaustion similar to cancer conditions. Given its success in treating cancer, anti-PD-1 / PD-L1 therapy has potential in the treatment of severe chronic infections. Furthermore, recent studies have shown that PD-1 regulates several metabolic changes in the development of T cell exhaustion.

[0129] Sepsis, associated with high mortality in intensive care units, is characterized by severe immunosuppression after an initial proinflammatory period. Postmortem examination of deceased septic patients highlights important immunological defects that impair host immunity. Sepsis has immunosuppressive mechanisms similar to those of cancer. Given the efficacy of CTLA-4 and PD-1 specific antibodies in improving host immunity and increasing survival in cancer patients, it is predicted that these agents will open promising avenues for the development of novel medicines against sepsis.

[0130] Working Example The following non-limiting examples are provided for illustrative purposes only to facilitate a more complete understanding of representative embodiments presently contemplated, and are not to be construed as limiting any of the embodiments described herein, including those relating to the compounds, pharmaceutical compositions, methods and uses described herein.

[0131] material and method Human peripheral blood pan-T cells were purchased from Stem Cell Technologies (Vancouver, BC, Canada) and Panc-1 cells were obtained from Kansas University Medical Center. 5-Fu, irinotecan, and oxaliplatin were purchased from Sigma (St. Louis, MO, USA). ImmunoCult-XF T Cell Exp Medium, ImmunoCult HuCD3 / CD28 / CD2 TCellAct, Hu Recom IL-2 (CHO Expr.) were purchased from Stem Cell Technologies (Vancouver, BC, Canada). ATPLite Luminescence Assay System and 96-well white, opaque, sterile tissue culture-treated microplates were purchased from Perkin Elmer (Waltham, Massachusetts, USA). RealTime Glo Annxin V Apoptosis and Annexin Assay was purchased from Promega (San Luis Obispo, CA, USA). Phospho-GSK-3β (Ser9), PD-L1 (E1L3N), PD-1 (D4W2J), β-actin, anti-rabbit IgG, HRP-linked antibody, Phospho-Akt (Thr308), Phospho-c-Jun (T-93) were purchased from Cell Signaling Technology (Danvers, Massachusetts, USA). 0.4 μm TC plate inserts with PC membrane 6-well plates were purchased from VWR (Radnor Pennsylvania, USA). APC-labeled PD-L1 (B7-H1) monoclonal antibody (MIH1) and FITC-labeled (PD-1) monoclonal antibody (MIH4) were purchased from Thermo Fisher Scientific (Waltham, Massachusetts, USA).

[0132] Compound Preparation: Stock solutions of the following compounds were prepared: 150 mM oxaliplatin in DMSO, 100 mM irinotecan in DMSO, 50 mM 5Fu in DMSO, and BCN057: 16 mg / mL BCN057 in 30% Captisol® (β-cyclodextrin sulfobutyl ether sodium).

[0133] Cell lines and cell culture: Panc-1 cells were cultured in DMEM medium supplemented with 10% fetal bovine serum and maintained at 37°C under 5% CO2. Human peripheral blood pan-T cells were cultured in serum-free T cell expansion medium with 10ng / ml rhIL-2 and expanded according to the manufacturer's instructions. Panc-1 cells and T cells were grown and expanded separately in T75 flasks. For co-culture experiments with Panc-1 cells, 2.5 million T cells per well were seeded into wells using 6-well plates. 2.5 million Panc-1 cells were seeded into thininserts, which were placed into wells of 6-well plates such that the cells were physically separated but shared medium. Cells were incubated at 37°C under 5% CO2 for 24 hours. At 24 hours, the medium was replaced with medium containing vehicle or 10uM BCN057 and incubated for the following time points: 1 hour, 6 hours, 12 hours, 24 hours. Panc-1 cells were removed by gently scraping the cells from the thinsert.

[0134] Flow Cytometry: Flow cytometric analysis: Panc-1 and T cells were harvested independently and washed with 1xPBS. Cells were washed 3xPBS and incubated with antibodies for 1 hour at room temperature. Cells were then washed 3xPBS, resuspended in 1xPBS and analyzed by flow cytometry using a NovoCyte flow cytometer (Agilent Technologies Inc. Santa Clara, CA) and data were analyzed with NovoExpress software.

[0135] Western Blot: Cells were harvested and lysed in cell lysis buffer for 15 min on ice, and lysates were centrifuged at 13,000 rpm, 4°C for 10 min. Supernatants were transferred to new tubes for BCA protein quantification assay (Thermo Fisher Scientific) and samples were normalized. 3x loading buffer (Cell Signaling) was added to protein samples and boiled for 5 min. Samples were used for SDS-PAGE analysis and transferred to nitrocellulose membranes. Membranes were blocked with 5% BSA for 1 h at room temperature and incubated with primary antibodies overnight at 4°C. Membranes were washed 3 times with 1xTBST for 5 min and incubated with horseradish peroxidase-labeled secondary antibodies for 1 h at room temperature. Proteins were visualized with SuperSignal West Pico Stable Peroxide Solution (Thermo Fisher Scientific). Blots were visualized by chemiluminescence using a BioRad Versadoc Imaging System.

[0136] Viability assay. To determine the effect of BCN057 compared to and in combination with irinotecan, oxaliplatin and 5FU, the ATPlite luminescence assay system was used according to the manufacturer's instructions. On day 1, 3,000 Panc-1 cells were seeded per well in 96-well white, opaque, sterile tissue culture treated microplates and incubated for 24 hours at 37°C, 5% CO2 in complete medium. On day 2, the medium was replaced with medium containing various combinations of 10 uM BCN057, 25 μM 5Fu, 75 μM oxaliplatin, and 50 μM irinotecan and incubated at 37°C, 5% CO2. On days 3, 4, and 5, the ATP Lite Assay was performed according to the manufacturer's instructions for 24, 48, 9, and 72 hours. Values ​​were expressed as a percentage of the vehicle control for each drug unless otherwise stated. No differences in survival rates were noted among either vehicle control.

[0137] QPCR: Compare the mRNA levels of BMI1 and LGR5 target genes in Panc-1 cells treated with BCN057 or vehicle. Quantitative PCR (qPCR) was used to measure the expression of stem cell marker LGR5, detected by real-time PCR using the primer pairs listed in Table 1, and BMI1. Total RNA was extracted using a Trizol kit (Invitrogen, CA, USA). [Table 1]

[0138] Statistics. Results are presented by calculating the mean (± standard deviation) relative to untreated controls, indicated with (n). Percent survival was calculated by dividing the mean of each condition by the mean of the untreated control (vehicle). Statistical analysis was performed in Excel.

[0139] Example 1 BCN057 inhibits pancreatic cancer cell proliferation by restoring apoptosis: The first example involved testing the effects of BCN057 in pancreatic cancer cells. Panc-1 cells, a G12D KRAS mutant cell line, were treated with BCN057 for 48 hours. The ATPlite™ assay (PerkinElmer) showed a significant decrease in the viability of Panc1 cells with increasing doses of BCN057.

[0140] Figure 1 is a histogram showing the effect of BCN057 on the survival of pancreatic cancer cell lines, assessed by ATP Lite™ assay (Perkin Elmer) with 72 hours of incubation after drug application. All data points are n=5. Controls (ND) received no drug and BCN057 was administered at final concentrations of 2, 5, 10, and 20 μM. Percent survival was calculated by dividing the mean value for each condition by the mean value (V) of control cells at each time point.

number

[0141] This data was analyzed using the Quest Graph™ IC 50 Plotted on the calculator, it is 1.33mm IC 50 This value was calculated as IC 50 This is comparable to other analyses of cytotoxic drugs such as gemcitabine against pancreatic ductal adenocarcinoma cells (AsPC-1, BxPC-3, MIA PaCa-2 and Panc-1) where the cytotoxicity ranged from 494 nM to 23.9 μM. The drug was found to exhibit potent cytotoxicity against the cells at all concentrations in the dose-response curve. Furthermore, these doses encompass the concentration range observed in plasma during rodent efficacy studies. Microscopic images then appeared to show cellular blebbing and fragmentation immediately after application of BCN057, indicating possible apoptosis. Figure 2A shows the time course of pancreatic cancer cell proliferation following treatment with 10 μM BCN057. Blebbing and cell fragmentation were observed within 20 min of incubation with the drug (indicated by arrows). Cell fragmentation was observed between 20 and 30 min after exposure to the drug.

[0142] Panc-1 cells harbor a heterozygous missense mutation at codon 12 (p.G12D; GGT>GAT of K-RAS) similar to other KRAS mutant tumor cells, and the phenotype is loss of apoptotic capacity and consequent chemoresistance. BCN057 treatment appeared to show rapid, dose-dependent restoration of apoptosis as soon as 20 min after exposure to the drug.

[0143] To verify the involvement of apoptosis in BCN057-induced cell death, Panc1 cells with and without BCN057 were subjected to Annexin V staining. Annexin V staining was performed up to 6 hours and measured as a function of BCN057 dose response. A dose- and time-dependent increase in apoptosis was detected in Panc1 cells treated with BCN057. Figure 2B is a graph of Annexin V apoptosis and necrosis assay in Panc-1 cells. Panc-1 cells were exposed to serial dilutions of BCN057 in the presence or absence of RealTime-Glo™ Annexin V apoptosis and necrosis assay reagent. Plates were incubated at 37°C / 5% CO2 and luminescence (Annexin V binding) was measured over a period of 6 hours. No changes were observed in fluorescence over the same period. Apoptosis appeared to increase with increasing concentrations of BCN057. Apoptosis was observed in a dose- and time-dependent manner at 10 min, 30 min, 1 h, and 6 h.

[0144] Further qPCR analysis was performed for Bax, Bcl-2, Akt-1 and caspase-3, which showed that BCN057 induced the expression of the pro-apoptotic gene Bax but suppressed the anti-apoptotic Bcl2 gene expression in Panc1 cells 1 hour after treatment. Figure 2C is a histogram of the results of qPCR analysis of pro- and anti-apoptotic markers in Panc-1 cells. BAX was significantly elevated and Bcl-2 was decreased 1 hour after exposure to BCN057. *p<0.05. The expression profile did not affect Akt-1 or caspase-3, consistent with caspase-independent cell death.

[0145] Example 2 BCN057 suppresses GSK3β signaling to restore apoptosis in PANC1 cells. The Panc1 cell line harbors a KRASG12D mutation and has lost the ability for apoptosis, which is characteristic of oncogenic RAS mutations. To explore the role that BCN057 may have in restoring apoptosis in this cell line, GSK3B c-Jun correlation was evaluated since it has been directly linked to apoptosis. Other investigators have reported an association of GSK3B with c-Jun and apoptosis, together with the fact that X is overexpressed in oncogenic KRAS phenotypes. Analysis of Panc1 cells for GSK3B inhibition over time after exposure to 10 μM BCN057 showed a rapid induction of phosphorylation at S-9 (Figure 3A and Figure 3B). This inhibition was also observed in other non-transformed cell types such as HEK298.

[0146] Phosphorylation of T91 / 93 in c-Jun was measured as a function of total c-Jun. Significant phosphorylation of T91 / 93 was observed, which has been reported to be associated with activation of the proapoptotic activity of c-Jun. This activity was not found to be induced in nontransformed cells treated with BCN057 or in nontransformed HEK298 epithelial cells (data not shown).

[0147] In KRAS-dependent tumors, c-Jun acts at the promoter level to suppress phosphatase and tensin homolog (PTEN) expression, and that suppression of PTEN expression is essential for anti-apoptosis and cell transformation by oncogenic RAS. Therefore, we investigated PTEN expression levels in response to BCN057. Consistent with our findings of c-Jun pro-apoptotic activation under BCN057 treatment of cells, we found increased expression of PTEN protein for up to 1 h (Figures 4B and 4C) followed by a decrease in cell apoptosis (Figure 2).

[0148] FIG. 4A is a Western blot of phospho-c-Jun (T-91 / 93). BCN057 significantly increases JNK-dependent phosphorylation in KRAS mutant Panc-1 cancer cells within 20 minutes of drug treatment. Phosphorylation of c-Jun at T-91 / 93 is known to induce apoptosis. Total c-Jun protein levels were unchanged. FIG. 4B shows PTEN expression in Panc-1 cells. In particular, the Western blot of PTEN shows protein expression over 24 hours with β-actin staining from the same samples with vehicle or BCN057 treatment (10 μM) at the indicated time points. FIG. 4C is a histogram of the ratio of densitometry values ​​of PTEN to β-actin showing the fold increase in PTEN levels in B. Arrows indicate the onset of apoptosis.

[0149] Example 3 BCN057 enhances the antitumor effects of chemotherapy agents. The anti-apoptotic phenotype of KRAS is associated with greater resistance to chemotherapy. Therefore, to investigate whether BCN057 potentiates cytotoxic chemotherapeutic agents such as 5-fluorouracil (also known as 5FU), Panc1 cells were pretreated with 5FU as reported and incubated with BCN057. The combination of 5FU and BCN057 at comparable concentrations and conditions showed greater cytotoxicity than 5FU alone (Figure 5A). The effect followed a dose-dependent response to BCN057 and was consistent over 24, 48, and 72 hours after BCN057 treatment (24 and 48 hours not shown).

[0150] FIG. 5A shows the effect of BCN057 and 5-FU on the survival of pancreatic cancer cell lines assessed by ATP Lite™ assay (Perkin Elmer) (all drugs were incubated for 72 hours). All data points are n=5. Controls (ND) received no drug. Test groups included 5FU (50 μM), 057 (BCN057: 2, 5, 10, and 20 μM) or 5FU+BCN057 (BCN057: 2, 5, 10, and 20 μM in combination with 50 μM 5FU). Percent survival was calculated by dividing the mean value for each condition by the mean value (V) including the standard deviation of the control cells at each time point.

[0151] LGR5 and BMI1 in pancreatic cancer: LGR5 showed widespread expression in all cells of pancreatic tumors. Others have employed BMI1 as an important contributor to the chemoresistance and invasiveness measurable in Panc-1. We tested whether BCN057 led to an expansion of these cell populations in Panc-1 cells, but no significant increase in abundance was observed at 10 μM (Figure 5B).

[0152] Epithelial Lgr5 stem cells: Previous reports from our group as well as other researchers have shown that Lgr5+ve crypt basal columnar cells are sensitive to chemoradiotherapy. We have also reported that protection of Lgr5+ve ISCs from treatment-induced toxicity is crucial to minimize intestinal epithelial damage. 5FU is a component of Forfirinox and is the active agent metabolized from capecitabine (Xeloda®). To investigate the effect of BCN057 on chemotherapy-induced toxicity in Lgr5+ISCs, Lgr5-Cre-ERT-GFP mice were pretreated with 5FU as reported and then subjected to BCN057 treatment. Confocal microscopic imaging of jejunal sections clearly showed a marked reduction in Lgr5+ISCs in 5FU-treated mice. However, 5FU-treated mice receiving BCN057 showed a prominent presence of Lgr5+ve ISCs (Figures 5C and 5D), suggesting that BCN057 protects ISCs from chemical toxicity similar to radiation.

[0153] Figure 5C shows that BCN057 protects GI epithelium and LGR5 stem cells (green) from 5FU cytotoxicity. GI crypt staining with green color = LGR5+ stem cells. Mice were treated with 5FU and 5FU+BCN057 for 5 days. BCN057 was administered for an additional 2 days after 5FU treatment. Small intestinal samples were collected and stained for LGR5 3 days after the last 5FU injection. Figure 5D is a histogram of % quantification of LGR5+ cells remaining in the 1 mM section of the crypt base, showing prevention of 5-FU-induced apoptosis in LGR5+ stem cells in the GI.

[0154] Combinations of chemotherapy and cytotoxicity: Other cytotoxic drugs commonly used in the treatment of pancreatic or colorectal cancer, such as those in Forfirinox, such as 5FU, irinotecan and oxaliplatin, were then investigated in a similar fashion in combination with equivalent doses of BCN057 for 24, 48 and 72 hours. In each case, restoration of apoptosis induced a statistically significant sensitivity to the cytotoxic effects of the drug or combination with BCN057 at all time points after treatment except one (irinotecan (IRI) and oxaliplatin (Ox) at 72 hours).

[0155] Figure 6 is a histogram showing the cytotoxicity of oxaliplatin and irinotecan, either individually or in combination with 5FU and BCN057, against Panc-1 cells. Cells were incubated with oxaliplatin, irinotecan and 5FU at a concentration of 10 μM, and also with BCN057 (057) at 10 μM, n=4 for all assays. In all cases, the addition of BCN057 led to a general trend of decreased tumor viability over 72 hours. *p<0.05, **p<0.005.

[0156] This trend remained the same in another study using much higher doses of oxaliplatin (75 μM), irinotecan and 5FU at concentrations of 50 μM and 10 μM, respectively, in BCN057 (057) (n=6, data not shown), and is consistent with the observations of others.

[0157] BCN057 and immune checkpoint blockade: An emerging approach to treatment for many forms of cancer, including PDAC, is immune checkpoint inhibitors. Since GSK3B is a key regulator of immune checkpoints, particularly in the PD-1 / PDL-1 axis, we sought to investigate the effect of BCN057 on PD-1 / PDL-1 expression. A culture system consisting of Panc1 cells and human T cells separated by a membrane was used as previously described (Neubert, NJ et al., Frontiers in Immunology 7, 2016). Panc-1 cells were co-cultured with T cells for 1 day and subsequently treated with 10 μM BCN057 for up to 12 hours.

[0158] Figure 7A is a Western blot showing PD-1 expression on human T lymphocytes co-cultured with Panc-1 cells for 1, 6 and 12 hours without drug (vehicle) or with BCN057. Figure 7B is a graph showing that inhibition of GSK3B induces lymphocyte survival upon exposure to radiation. In vitro viability assays using increasing concentrations of BCN057 on irradiated mouse T lymphocytes showed EC 50 = 5.34 mM, which is within the observed plasma concentrations for the drug.

[0159] Figure 8 is a Western blot showing PDL-1 expression (red arrow) on Panc-1 cells co-cultured with human T lymphocytes for 2.5, 5, 7.5 and 10 hours without drug (vehicle) or with BCN057. PDL-1 expression on Panc-1 cells did not change appreciably, but the cells underwent rapid apoptosis, as described. However, PD-1 expression on human T lymphocytes was markedly decreased within the first hour and persisted for 12 hours after treatment, as shown in Figure 7A. In another study with mouse T lymphocytes, 2 Gy irradiation was used to induce apoptosis with increasing doses of BCN057. The results showed that EC 50= 5.34 μM (Figure 7B), thus establishing a difference in the biological response between tumor and immune cells.

[0160] Taken together, the results show that BCN057 can selectively enhance the cytotoxic effects of chemotherapeutic agents in oncogenic KRAS pancreatic cancer cells. Furthermore, it mitigates the toxic effects of chemotherapeutic agents in normal intestinal epithelium, suggesting that BCN057 can be considered as an adjunct therapy to improve the therapeutic index. As an agent that can restore apoptosis in oncogenic RAS phenotypes, this fits nicely with the use case to induce greater tumor sensitivity to chemotherapeutic agents. In a previous paper (Bhanja, P., et.al., Stem Cell Research & Therapy 9, 2018), we demonstrated a mouse model using a syngeneic KRASG13R mutant MC38 tumor cell line, in which mice receiving BCN057 had significantly lower tumor burdens than controls, but mice receiving BCN057 with radiation therapy had lower tumor burdens and were able to withstand much higher radiation doses than mice receiving radiation alone. This highlights the value of targeting pathways that are so differentially connected in transformed versus non-transformed phenotypes, and this dual activity in tumoricidal as well as normal tissue sparing activity may result in better overall and progression-free survival.

[0161] Pharmaceutical Carriers and Administration The method of the present invention can prevent disease or condition or one or more symptoms of disease or condition.As used herein, a therapeutic agent that "prevents" a disorder or condition refers to a compound that reduces the occurrence of a disorder or condition in treated samples compared to untreated control samples in statistical samples, or delays the onset or reduces the severity of one or more symptoms of a disorder or condition compared to untreated control samples.

[0162] The compositions and methods of the present disclosure may be used to treat individuals in need thereof.In certain embodiments, the individual is a mammal, such as a human or non-human mammal.When administered to an animal, such as a human, the composition or compound is preferably administered or used as a pharmaceutical composition, for example, comprising the compound of the present disclosure and a pharmaceutically acceptable carrier.

[0163] Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil, or injectable organic esters. In a preferred embodiment, when such pharmaceutical compositions are for administration to humans, especially for invasive routes of administration (i.e., routes such as injection or implantation that avoid transport or diffusion through epithelial barriers), the aqueous solutions are pyrogen-free or substantially pyrogen-free. Excipients can be selected, for example, to effect delayed release of the drug or to selectively target one or more cells, tissues, or organs. Pharmaceutical compositions can be in dosage unit form, such as tablets, capsules (including sprinkle capsules and gelatin capsules), granules, lyophilized formulations for reconstitution, powders, liquids, syrups, suppositories, injections, and the like. The compositions can also be present in transdermal delivery systems, for example, skin patches.

[0164] The pharmaceutical composition disclosed herein may contain a sufficient amount of therapeutic compound to allow daily administration to an individual.In certain embodiments, the pharmaceutical composition disclosed herein may contain, for example, at least 5mg, at least 10mg, at least 15mg, at least 20mg, at least 25mg, at least 30mg, at least 35mg, at least 40mg, at least 45mg, at least 50mg, at least 55mg, at least 60mg, at least 65mg, at least 70mg, at least 75mg, at least 80mg, at least 85mg, at least 90mg, at least 95mg, or at least 100mg of therapeutic compound. In certain embodiments, a pharmaceutical composition disclosed herein may contain, for example, at least 5 mg, at least 10 mg, at least 20 mg, at least 25 mg, at least 50 mg, at least 75 mg, at least 100 mg, at least 200 mg, at least 300 mg, at least 400 mg, at least 500 mg, at least 600 mg, at least 700 mg, at least 800 mg, at least 900 mg, at least 1,000 mg, at least 1,100 mg, at least 1,200 mg, at least 1,300 mg, at least 1,400 mg, or at least 1,500 mg of a therapeutic compound. In yet another aspect of this embodiment, the pharmaceutical compositions disclosed herein may contain, for example, in the range of about 5 mg to about 100 mg, about 10 mg to about 100 mg, about 50 mg to about 150 mg, about 100 mg to about 250 mg, about 150 mg to about 350 mg, about 250 mg to about 500 mg, about 350 mg to about 600 mg, about 500 mg to about 750 mg, about 600 mg to about 900 mg, about 750 mg to about 1,000 mg, about 850 mg to about 1,200 mg, or about 1,000 mg to about 1,500 mg.In further specific embodiments, the pharmaceutical compositions disclosed herein may be administered in an amount of, for example, about 10 mg to about 250 mg, about 10 mg to about 500 mg, about 10 mg to about 750 mg, about 10 mg to about 1,000 mg, about 10 mg to about 1,500 mg, about 50 mg to about 250 mg, about 50 mg to about 500 mg, about 50 mg to about 750 mg, about 50 mg to about 1,000 mg, about 50 mg to about 1,500 mg, about 10 ... It may contain in the range of about 250 mg, about 100 mg to about 500 mg, about 100 mg to about 750 mg, about 100 mg to about 1,000 mg, about 100 mg to about 1,500 mg, about 200 mg to about 500 mg, about 200 mg to about 750 mg, about 200 mg to about 1,000 mg, about 200 mg to about 1,500 mg, about 5 mg to about 1,500 mg, about 5 mg to about 1,000 mg, or about 5 mg to about 250 mg.

[0165] The pharmaceutical compositions disclosed herein may include a solvent, emulsion, or other diluent in an amount sufficient to dissolve the therapeutic compounds disclosed herein. In certain embodiments, the pharmaceutical compositions disclosed herein may include a solvent, emulsion, or diluent in an amount of, for example, less than about 90% (v / v), less than about 80% (v / v), less than about 70% (v / v), less than about 65% (v / v), less than about 60% (v / v), less than about 55% (v / v), less than about 50% (v / v), less than about 45% (v / v), less than about 40% (v / v), less than about 35% (v / v), less than about 30% (v / v), less than about 25% (v / v), less than about 20% (v / v), less than about 15% (v / v), less than about 10% (v / v), less than about 5% (v / v), or less than about 1% (v / v). In certain embodiments, the pharmaceutical compositions disclosed herein contain a solvent, emulsion or other diluent in an amount of, for example, about 1% (v / v) to 90% (v / v), about 1% (v / v) to 70% (v / v), about 1% (v / v) to 60% (v / v), about 1% (v / v) to 50% (v / v), about 1% (v / v) to 40% (v / v), about 1% (v / v) to 30% (v / v), or about 1% (v / v) to 50% (v / v). %(v / v), approx. 1%(v / v)~20%(v / v), approx. 1%(v / v)~10%(v / v), approx. 2%(v / v)~50%(v / v), approx. 2%(v / v)~40 %(v / v), approx. 2%(v / v)~30%(v / v), approx. 2%(v / v)~20%(v / v), approx. 2%(v / v)~10%(v / v), approx. 4%(v / v)~50% (v / v), approx. 4%(v / v)~40%(v / v), approx. 4%(v / v)~30%(v / v), approx. 4%(v / v)~20%(v / v), approx. 4%(v / v)~10% (v / v), approximately 6%(v / v)~50%(v / v), approximately 6%(v / v)~40%(v / v), approximately 6%(v / v)~30%(v / v), approximately 6%(v / v)~20%( v / v), about 6% (v / v) to 10% (v / v), about 8% (v / v) to 50% (v / v), about 8% (v / v) to 40% (v / v), about 8% (v / v) to 30% (v / v), about 8% (v / v) to 20% (v / v), about 8% (v / v) to 15% (v / v), or about 8% (v / v) to 12% (v / v).

[0166] The final concentration of the therapeutic compound disclosed herein in the pharmaceutical composition disclosed herein can be any suitable concentration.In certain embodiments, the final concentration of the therapeutic compound in the pharmaceutical composition can be a therapeutically effective amount.In certain embodiments, the final concentration of the therapeutic compound in the pharmaceutical composition can be, for example, at least 0.00001mg / mL, at least 0.0001mg / mL, at least 0.001mg / mL, at least 0.01mg / mL, at least 0.1mg / mL, at least 1mg / mL, at least 10mg / mL, at least 25mg / mL, at least 50mg / mL, at least 100mg / mL, at least 200mg / mL, at least 500mg / mL, at least 700mg / mL, at least 1,000mg / mL, or at least 1,200mg / mL. In certain embodiments, the concentration of the therapeutic compound in the solutions disclosed herein may be, for example, up to 1,000 mg / mL, up to 1,100 mg / mL, up to 1,200 mg / mL, up to 1,300 mg / mL, up to 1,400 mg / mL, up to 1,500 mg / mL, up to 2,000 mg / mL, up to 2,000 mg / mL, or up to 3,000 mg / mL. In certain embodiments, the final concentration of the therapeutic compound in the pharmaceutical composition may be, for example, about 0.00001 mg / mL to about 3,000 mg / mL, about 0.0001 mg / mL to about 3,000 mg / mL, about 0.01 mg / mL to about 3,000 mg / mL, about 0.1mg / mL to approx. 3,000mg / mL, approx. 1mg / mL to approx. 3,000mg / mL, approx. 250mg / mL to approx. 3,000mg / mL, approx. 500mg / mL to approx. 3,000mg / mL, approx. 750mg / mL to approx. 3,000mg / mL, approx. 1,000m g / mL ~ approx. 3,000 mg / mL, approx. 100 mg / mL ~ approx. 2,000 mg / mL, approx. 250 mg / mL ~ approx. 2,000 mg / mL, approx. 500 mg / mL ~ approx. 2,000 mg / mL, approx. 750 mg / mL ~ approx. 2,000 mg / mL, approx. 1,000 m g / mL~about 2,000mg / mL, about 100mg / mL~about 1,500mg / mL, about 250mg / mL~about 1,500mg / mL, about 500mg / mL~about 1,500mg / mL, about 750mg / mL~about 1,500mg / mL, about 1,000m g / mL ~ approx. 1,500 mg / mL, approx. 100 mg / mL ~ approx. 1,200 mg / mL, approx. 250 mg / mL ~ approx. 1,200 mg / mL, approx. 500 mg / mL ~ approx. 1,200 mg / mL, approx. 750 mg / mL ~ approx. 1,200 mg / mL, approx. 1,000 m g / mL~about 1,200mg / mL, about 100mg / mL~about 1,000mg / mL, about 250mg / mL~about 1,000mg / mL, about 500mg / mL~about 1,000mg / mL, about 750mg / mL~about 1,000mg / mL, about 100mg / mL mL ~ approx. 750 mg / mL, approx. 250 mg / mL ~ approx. 750 mg / mL, approx. 500 mg / mL ~ approx. 750 mg / mL, approx. 100 mg / mL ~ approx. 500 mg / mL, approx. 250 mg / mL ~ approx. 500 mg / mL, approx. 0.00001 mg / mL ~ approx. 0.00 The concentration may be in the range of about 0.01 mg / mL, about 0.00001 mg / mL to about 0.001 mg / mL, about 0.00001 mg / mL to about 0.01 mg / mL, about 0.00001 mg / mL to about 0.1 mg / mL, about 0.00001 mg / mL to about 1 mg / mL, about 0.001 mg / mL to about 0.01 mg / mL, about 0.001 mg / mL to about 0.1 mg / mL, about 0.001 mg / mL to about 1 mg / mL, about 0.001 mg / mL to about 10 mg / mL, or about 0.001 mg / mL to about 100 mg / mL.

[0167] In certain embodiments, the therapeutically effective amount of the therapeutic compounds disclosed herein typically ranges from about 0.001 mg / kg / day to about 100 mg / kg / day. In certain embodiments, the therapeutically effective amount of the therapeutic compounds disclosed herein may be, for example, at least 0.001 mg / kg / day, at least 0.01 mg / kg / day, at least 0.1 mg / kg / day, at least 1.0 mg / kg / day, at least 5.0 mg / kg / day, at least 10 mg / kg / day, at least 15 mg / kg / day, at least 20 mg / kg / day, at least 25 mg / kg / day, at least 30 mg / kg / day, at least 35 mg / kg / day, at least 40 mg / kg / day, at least 45 mg / kg / day, or at least 50 mg / kg / day.In certain embodiments, an effective amount of a therapeutic compound disclosed herein can be, for example, about 0.001 mg / kg / day to about 10 mg / kg / day, about 0.001 mg / kg / day to about 15 mg / kg / day, about 0.001 mg / kg / day to about 20 mg / kg / day, about 0.001 mg / kg / day to about 25 mg / kg / day, about 0.001 mg / kg / day to about 30 mg / kg / day, about 0.001 mg / kg / day to about 35 mg / kg / day, about 0.001 mg / kg / day to about 40 mg / kg / day, about 0.001 mg / kg / day to about 50 mg / kg / day, about 0.001 mg / kg / day to about 60 mg / kg / day, about 0.001 mg / kg / day to about 70 mg / kg / day, about 0.001 mg / kg / day to about 80 mg / kg / day, about 0.001 mg / kg / day to about 100 mg / kg / day, about 0.001 mg / kg / day to about 150 mg / kg / day, about 0.001 mg / kg / day to about 20 mg / kg / day, about 0.001 mg / kg / day to about 25 mg / kg / day, about 0.001 mg / kg / day to about 30 mg / kg / day, about 0.001 mg / kg / day to about 150 mg / kg / day, about 0.001 mg / kg / day to about 150 mg / kg / day, about 0.001 mg / kg / day to about 20 mg / kg / day, about 0.001 mg / kg / day to about 25 mg / kg / day, about 0.001 mg / kg / day to about 30 mg / kg / day, about 0.0 / day ~ approx. 35 mg / kg / day, approx. 0.001 mg / kg / day ~ approx. 40 mg / kg / day, approx. 0.001 mg / kg / day ~ approx. 45 mg / kg / day, approx. 0.001 mg / kg / day ~ approx. 50 mg / kg / day, about 0.001mg / kg / day to about 75mg / kg / day, about 0.001mg / kg / day to about 100mg / kg / day, about 0.001mg / kg / day to about 150mg / kg / day, about 0.001 mg / kg / day ~ approx. 200 mg / kg / day, approx. 0.001 mg / kg / day ~ approx. 250 mg / kg / day, approx. 0.001 mg / kg / day ~ approx. 300 mg / kg / day, approx. 0.001 mg / kg / day ~ Approximately 350mg / kg / day, approximately 0.001mg / kg / day to approximately 400mg / kg / day, approximately 0.001mg / kg / day to approximately 450mg / kg / day, approximately 0.001mg / kg / day to approximately 500mg / k g / day, about 0.001 mg / kg / day to about 550 mg / kg / day, about 0.001 mg / kg / day to about 600 mg / kg / day, about 0.001 mg / kg / day to about 650 mg / kg / day, about 0.001 mg / kg / day to about 700 mg / kg / day, about 0.001 mg / kg / day to about 750 mg / kg / day, or about 0.001 mg / kg / day to about 800 mg / kg / day.In yet another aspect of this embodiment, an effective amount of a therapeutic compound disclosed herein can be, for example, about 0.01 mg / kg / day to about 10 mg / kg / day, about 0.01 mg / kg / day to about 15 mg / kg / day, about 0.01 mg / kg / day to about 20 mg / kg / day, about 0.01 mg / kg / day to about 25 mg / kg / day, about 0.01 mg / kg / day to about 30 mg / kg / day, about 0.01 mg / kg / day to about 35 mg / kg / day, about 0.01 mg / kg / day to about 40 mg / kg / day, about 0.01 mg / kg / day to about 50 mg / kg / day, about 0.01 mg / kg / day to about 55 mg / kg / day, about 0.01 mg / kg / day to about 60 mg / kg / day, about 0.01 mg / kg / day to about 70 mg / kg / day, about 0.01 mg / kg / day to about 80 mg / kg / day, about 0.01 mg / kg / day to about 90 mg / kg / day, about 0.01 mg / kg / day to about 100 mg / kg / day, about 0.01 mg / kg / day to about 150 mg / kg / day, about 0.01 mg / kg / day to about 20 mg / kg / day, about 0.01 mg / kg / day to about 25 mg / kg / day, about 0.01 mg / kg / day to about 30 mg / kg / day, about 0.01 mg / kg / day to about 150 mg / kg / day, about 0.01 mg / kg / day to about 150 mg / kg / day, about 0.01 mg / kg / day to about 20 mg / kg / day, about 0.01 mg / kg / day to about 25 mg / kg / day, mg / kg / day ~ approx. 35 mg / kg / day, approx. 0.01 mg / kg / day ~ approx. 40 mg / kg / day, approx. 0.01 mg / kg / day ~ approx. 45 mg / kg / day, approx. 0.01 mg / kg / day ~ approx. 50 mg / kg / day, about 0.01 mg / kg / day to about 75 mg / kg / day, about 0.01 mg / kg / day to about 100 mg / kg / day, about 0.01 mg / kg / day to about 150 mg / kg / day, about 0 .01mg / kg / day ~ approx. 200mg / kg / day, approx. 0.01mg / kg / day ~ approx. 250mg / kg / day, approx. 0.01mg / kg / day ~ approx. 300mg / kg / day, approx. 0.01mg / kg / day ~ approx. 350mg / kg / day, approx. 0.01mg / kg / day ~ approx. 400mg / kg / day, approx. 0.01mg / kg / day ~ approx. 450mg / kg / day, approx. 0.01mg / kg / day ~ approx. 500mg / kg / day, about 0.01 mg / kg / day to about 550 mg / kg / day, about 0.01 mg / kg / day to about 600 mg / kg / day, about 0.01 mg / kg / day to about 650 mg / kg / day, about 0.01 mg / kg / day to about 700 mg / kg / day, about 0.01 mg / kg / day to about 750 mg / kg / day, or about 0.01 mg / kg / day to about 800 mg / kg / day.In certain embodiments, an effective amount of a therapeutic compound disclosed herein can be, for example, about 0.1 mg / kg / day to about 10 mg / kg / day, about 0.1 mg / kg / day to about 15 mg / kg / day, about 0.1 mg / kg / day to about 20 mg / kg / day, about 0.1 mg / kg / day to about 25 mg / kg / day, about 0.1 mg / kg / day to about 30 mg / kg / day, about 0.1 mg / kg / day to about 35 mg / kg / day, about 0.1 mg / kg / day to about 40 mg / kg / day, about 0.1 mg / kg / day to about 50 mg / kg / day, about 0.1 mg / kg / day to about 60 mg / kg / day, about 0.1 mg / kg / day to about 70 mg / kg / day, about 0.1 mg / kg / day to about 80 mg / kg / day, about 0.1 mg / kg / day to about 90 mg / kg / day, about 0.1 mg / kg / day to about 100 mg / kg / day, about 0.1 mg / kg / day to about 150 mg / kg / day, about 0.1 mg / kg / day to about 20 mg / kg / day, about 0.1 mg / kg / day to about 25 mg / kg / day, about 0.1 mg / kg / day to about 30 mg / kg / day, about 0.1 mg / kg / day to about 15 ... / day ~ approx. 35mg / kg / day, approx. 0.1mg / kg / day ~ approx. 40mg / kg / day, approx. 0.1mg / kg / day ~ approx. 45mg / kg / day, approx. 0.1mg / kg / day ~ approx. 50mg / k g / day, about 0.1 mg / kg / day to about 75 mg / kg / day, about 0.1 mg / kg / day to about 100 mg / kg / day, about 0.1 mg / kg / day to about 150 mg / kg / day, about 0.1 m g / kg / day ~ approx. 200 mg / kg / day, approx. 0.1 mg / kg / day ~ approx. 250 mg / kg / day, approx. 0.1 mg / kg / day ~ approx. 300 mg / kg / day, approx. 0.1 mg / kg / day ~ Approximately 350mg / kg / day, approximately 0.1mg / kg / day to approximately 400mg / kg / day, approximately 0.1mg / kg / day to approximately 450mg / kg / day, approximately 0.1mg / kg / day to approximately 500mg / k g / day, about 0.1 mg / kg / day to about 550 mg / kg / day, about 0.1 mg / kg / day to about 600 mg / kg / day, about 0.1 mg / kg / day to about 650 mg / kg / day, about 0.1 mg / kg / day to about 700 mg / kg / day, about 0.1 mg / kg / day to about 750 mg / kg / day, or about 0.1 mg / kg / day to about 800 mg / kg / day.In certain embodiments, an effective amount of a therapeutic compound disclosed herein can be, for example, about 10 mg / kg / day to about 15 mg / kg / day, about 10 mg / kg / day to about 20 mg / kg / day, about 10 mg / kg / day to about 25 mg / kg / day, about 10 mg / kg / day to about 30 mg / kg / day, about 10 mg / kg / day to about 35 mg / kg / day, about 10 mg / kg / day to about 40 mg / kg / day, about 10 mg / kg / day to about 45 mg / kg / day, about 10 mg / kg / day to about 50 mg / kg / day, about 10 mg / kg / day to about 75 mg / kg / day, about 10 mg / kg / day to about 100 mg / kg / day, about 10 mg / kg / day to about 150 mg / kg / day, about 10 mg / kg / day to about 200 mg / kg / day, or about 10 mg / kg / day to about 300 mg / kg / day. g / kg / day, about 10 mg / kg / day to about 250 mg / kg / day, about 10 mg / kg / day to about 300 mg / kg / day, about 10 mg / kg / day to about 350 mg / kg / day, about 10 mg / kg / day to about 400 mg / kg / day, about 10 mg / kg / day to about 450 mg / kg / day, about 10 mg / kg / day to about 500 mg / kg / day, about 10 mg / kg / day to about 550 mg / kg / day, about 10 mg / kg / day to about 600 mg / kg / day, about 10 mg / kg / day to about 650 mg / kg / day, about 10 mg / kg / day to about 700 mg / kg / day, about 10 mg / kg / day to about 750 mg / kg / day, or about 10 mg / kg / day to about 800 mg / kg / day.

[0168] In other aspects of this embodiment, an effective amount of a therapeutic compound disclosed herein may be in the range of, for example, about 1 mg / kg / day to about 10 mg / kg / day, about 1 mg / kg / day to about 15 mg / kg / day, about 1 mg / kg / day to about 20 mg / kg / day, about 1 mg / kg / day to about 25 mg / kg / day, about 1 mg / kg / day to about 30 mg / kg / day, about 1 mg / kg / day to about 35 mg / kg / day, about 1 mg / kg / day to about 40 mg / kg / day, about 1 mg / kg / day to about 45 mg / kg / day, about 1 mg / kg / day to about 50 mg / kg / day, about 1 mg / kg / day to about 75 mg / kg / day, or about 1 mg / kg / day to about 100 mg / kg / day. In certain embodiments, an effective amount of a therapeutic compound disclosed herein may range, for example, from about 5 mg / kg / day to about 10 mg / kg / day, from about 5 mg / kg / day to about 15 mg / kg / day, from about 5 mg / kg / day to about 20 mg / kg / day, from about 5 mg / kg / day to about 25 mg / kg / day, from about 5 mg / kg / day to about 30 mg / kg / day, from about 5 mg / kg / day to about 35 mg / kg / day, from about 5 mg / kg / day to about 40 mg / kg / day, from about 5 mg / kg / day to about 45 mg / kg / day, from about 5 mg / kg / day to about 50 mg / kg / day, from about 5 mg / kg / day to about 75 mg / kg / day, or from about 5 mg / kg / day to about 100 mg / kg / day.

[0169] In liquid and semi-solid formulations, the concentration of the therapeutic compounds disclosed herein may typically be from about 50 mg / mL to about 1,000 mg / mL. In certain embodiments, the therapeutically effective amount of the therapeutic compounds disclosed herein may be, for example, from about 50 mg / mL to about 100 mg / mL, from about 50 mg / mL to about 200 mg / mL, from about 50 mg / mL to about 300 mg / mL, from about 50 mg / mL to about 400 mg / mL, from about 50 mg / mL to about 500 mg / mL, from about 50 mg / mL to about 600 mg / mL, from about 50 mg / mL to about 700 mg / mL, from about 50 mg / mL to about 800 mg / mL, from about 50 mg / mL to about 900 mg / mL, from about 50 mg / mL to about 1,000 mg / mL, or from about 100 mg / mL to about 200 mg / mL. 00mg / mL~about 200mg / mL, about 100mg / mL~about 300mg / mL, about 100mg / mL~about 400mg / mL, about 100mg / mL~about 500mg / mL, about 100mg / mL~about 600mg / mL, about 100mg / mL~about 700mg / m L, about 100mg / mL to about 800mg / mL, about 100mg / mL to about 900mg / mL, about 100mg / mL to about 1,000mg / mL, about 200mg / mL to about 300mg / mL, about 200mg / mL to about 400mg / mL, about 200mg / mL to about 50 0mg / mL, about 200mg / mL to about 600mg / mL, about 200mg / mL to about 700mg / mL, about 200mg / mL to about 800mg / mL, about 200mg / mL to about 900mg / mL, about 200mg / mL to about 1,000mg / mL, about 300mg / mL mL~about 400mg / mL, about 300mg / mL~about 500mg / mL, about 300mg / mL~about 600mg / mL, about 300mg / mL~about 700mg / mL, about 300mg / mL~about 800mg / mL, about 300mg / mL~about 900mg / mL, about 300 mg / mL~about 1,000mg / mL, about 400mg / mL~about 500mg / mL, about 400mg / mL~about 600mg / mL, about 400mg / mL~about 700mg / mL, about 400mg / mL~about 800mg / mL, about 400mg / mL~about 900mg / m L, about 400mg / mL to about 1,000mg / mL, about 500mg / mL to about 600mg / mL, about 500mg / mL to about 700mg / mL, about 500mg / mL to about 800mg / mL, about 500mg / mL to about 900mg / mL, about 500mg / mL to about 1,000 mg / mL, about 600 mg / mL to about 700 mg / mL, about 600 mg / mL to about 800 mg / mL, about 600 mg / mL to about 900 mg / mL, or about 600 mg / mL to about 1,000 mg / mL.

[0170] The subject may be a human, rat, mouse, cat, dog, horse, sheep, cow, monkey, bird, or amphibian. In other embodiments, the cells are in vivo or in vitro. Typical subjects to which the compounds of the present disclosure may be administered are mammals, particularly primates. For veterinary applications, a wide variety of subjects are suitable, such as livestock such as cows, sheep, goats, cows, pigs, poultry such as chickens, ducks, geese, turkeys, and pets such as livestock, particularly dogs and cats. For diagnostic or research applications, a wide variety of mammals are suitable subjects, including rodents (e.g., mice, rats, hamsters), rabbits, primates, and pigs, such as inbred pigs. Furthermore, for in vitro applications, such as in vitro diagnostic and research applications, mammals, particularly primates, such as human blood, urine, or tissue samples, such as body fluids and cell samples of the subjects, or blood urine or tissue samples of animals described for veterinary applications, are suitable for use.

[0171] When administered to an organism, the compound may be administered by any suitable means. In some embodiments, the compound or formulation is administered orally. In some embodiments, the compound or formulation is administered by injection, for example, subcutaneously, parenterally, or intravenously.

[0172] In some embodiments, the compound may be administered in combination with other possible alleviators. In certain embodiments, the composition may be administered with growth factors, NSAIDs, chemotherapeutic agents, anti-inflammatory agents, antibiotics, metformin (Lucophage, Glumetzer, etc.), sulfonylureas, meglitinides, thiazolidinediones, DPP-4 inhibitors, GLP-1 receptor agonists, SGLT2 inhibitors, and / or insulin therapy agents for the treatment of the above conditions. In one aspect, the growth factor may be G-CSF (also known as filgrastim, NEUPOGEN®) or erythropoietin (also known as Epogen®).

[0173] In other embodiments, the composition may include an effective amount of the modulator and / or other pharma- ceutical active agent in a physiologically acceptable carrier. The carrier may be in a wide variety of forms depending on the form of formulation desired for a particular route of administration. Suitable carriers and their formulations are described, for example, in Remington's Pharmaceutical Sciences, by EW Martin. In some embodiments, the compound may be included in any suitable amount in any suitable carrier material, and is typically present in an amount of 1-95% by weight of the total weight of the composition. The composition may be provided in a dosage form suitable for parenteral (e.g., subcutaneous, intravenous, intramuscular, or intraperitoneal) or oral routes of administration. Pharmaceutical compositions may be formulated according to conventional pharmaceutical practice (see, for example, Remington: The Science and Practice of Pharmacy (20th ed.), ed. A. R. Gennaro, Lippincott Williams & Wilkins, 2000 and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York).

[0174] In some embodiments, the composition may be in a form suitable for administration by sterile injection. In one example, to prepare such a composition, the composition is dissolved or suspended in a parenterally acceptable liquid vehicle. Particularly acceptable vehicles and solvents that may be used are water, water adjusted to a suitable pH by adding an appropriate amount of hydrochloric acid, sodium hydroxide or a suitable buffer, 1,3-butanediol, Ringer's solution and isotonic sodium chloride and dextrose solution. Aqueous preparations may also contain one or more preservatives (e.g., methyl, ethyl or propyl p-oxybenzoate). For parenteral preparations, the carrier usually comprises sterile water, but may contain other ingredients, such as ingredients that aid solubility or for preservation. Injectable suspensions can also be prepared, in which case suitable stabilizers may be used. In one embodiment, the formulation includes at least one or more of methanesulfonic acid, povidone, benzyl alcohol, n-methylpyrrolidone, ethanol, poloxamer 188, lactic acid, Captisol (SBE-beta-CD), or Vitamin E, such as TPGS (d-alpha tocopheryl polyethylene glycol 1000 succinate).

[0175] Formulations suitable for parenteral administration usually include a sterile aqueous preparation of the compound, which is isotonic with the blood of the recipient (e.g., physiological saline). Such preparations may include suspending agents and thickening agents, as well as liposomes or other microparticulate systems designed to target the compound to blood components or one or more organs. The formulations may be administered in single or multiple dose forms.

[0176] Parenteral administration can include any form suitable for systemic or local delivery. Administration can be, for example, intravenous, intraarterial, intrathecal, intramuscular, subcutaneous, intramuscular, intraperitoneal (e.g., intraperitoneal), etc., and can be performed by infusion pump (external or implantable) or any other means suitable for the desired mode of administration.

[0177] In some embodiments, the composition may be in a form suitable for oral administration. For the composition in oral dosage form, any of the usual pharmaceutical media may be used. Thus, for example, for liquid oral preparations such as suspensions, elixirs and solutions, suitable carriers and additives include water, glycols, oils, alcohols, flavorings, preservatives, colorings, etc. For example, for solid oral preparations such as powders, capsules and tablets, suitable carriers and additives include starches, sugars, diluents, granulating agents, lubricants, binders, disintegrating agents, etc. If desired, tablets may be sugar-coated or enteric-coated by standard techniques.

[0178] Compositions suitable for oral administration (e.g., by ingestion) may be provided as discrete units, such as capsules, cachets, tablets, or lozenges, each containing a predetermined amount of the active ingredient as a powder or granules. In some cases, suspensions in aqueous or non-aqueous solutions, such as syrups, elixirs, emulsions, or liquids, may be employed. Formulations for oral use include tablets containing the active ingredient in a mixture with pharma- ceutically acceptable excipients. Such formulations are known to those skilled in the art. Excipients may be, for example, inert diluents or fillers (e.g., sucrose, sorbitol, cane sugar, mannitol, microcrystalline cellulose, starch including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate); granulating and disintegrating agents (e.g., microcrystalline cellulose, starch cellulose derivatives, starch including potato starch, croscarmellose sodium, sodium alginate, or alginic acid); binders (e.g., sucrose, glucose, sorbitol, gum arabic, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethylene glycol); and lubricants, glidants, and antiadhesives (e.g., magnesium stearate, zinc stearate, stearic acid, silica, hydrogenated vegetable oil, or talc). Other pharma- ceutically acceptable excipients can be colorants, flavoring agents, plasticizers, humectants, buffering agents, and the like.

[0179] Syrups can be made by adding the compound to a concentrated aqueous solution of sugar, e.g., sucrose, to which any auxiliary ingredients may also be added, which may include flavorings, suitable preservatives, agents that retard the crystallization of sucrose, and agents that increase the solubility of any other ingredients, such as polyhydric alcohols, e.g., glycerol or sorbitol.

[0180] In some embodiments, the composition may be in the form of a nasal or other mucous membrane spray formulation (e.g., inhalable form). These formulations may include purified aqueous solutions of active compounds, including preservatives and isotonicity agents. Such formulations may be adjusted to a pH and isotonic state compatible with nasal or other mucous membranes. Alternatively, the formulation may be in the form of a finely divided solid powder suspended in a gas carrier. Such formulations may be delivered by any suitable means or method, for example, by a nebulizer, atomizer, metered dose inhaler, etc.

[0181] In some embodiments, the compositions may be in a form suitable for rectal administration. These preparations may be provided as a suppository with a suitable carrier such as cocoa butter, hydrogenated fats or hydrogenated fatty carboxylic acids.

[0182] In some embodiments, the composition may be in a form suitable for transdermal administration.These formulations may be prepared, for example, by incorporating the active compound into a thixotropic or gelatinous carrier such as a cellulose-based medium, for example, methylcellulose or hydroxyethylcellulose, and then loading the resulting formulation into a transdermal device adapted to be fixed in transdermal contact with the skin of a wearer.

[0183] In addition to the components described above, the compositions of the present disclosure may further comprise one or more accessory ingredients selected from encapsulating materials, diluents, buffers, flavoring agents, binders, disintegrants, surfactants, thickeners, lubricants, preservatives (including antioxidants), and the like.

[0184] In some embodiments, the composition may be formulated for immediate release, sustained release, delayed onset release or any other release profile known to those skilled in the art. In some embodiments, the pharmaceutical composition may be formulated to release the active compound substantially immediately upon administration or at any predetermined time or period after administration. The latter type of composition is generally known as a controlled release formulation, and these include: (i) formulations that create a substantially constant concentration of drug in the body for an extended period of time; (ii) formulations that create a substantially constant concentration of drug in the body for an extended period of time after a pre-determined lag time; (iii) formulations that maintain a relatively constant effective level in the body to maintain a predetermined period of action while simultaneously minimizing undesirable side effects associated with fluctuations in the plasma concentration of the active substance (sawtooth kinetic pattern); (iv) formulations that localize the action, for example, by spatial location of the controlled release composition adjacent to or within the central nervous system or cerebrospinal fluid; (v) formulations that allow convenient administration, such as a dose being administered once every one or two weeks; and (vi) formulations that target the site of the pathology. For some applications, controlled release formulations obviate the need for frequent administration to maintain activity at medically beneficial levels.

[0185] Any of several strategies can be pursued to obtain controlled release, where the release rate exceeds the metabolism rate of the compound.In one example, controlled release can be obtained by appropriate selection of various formulation parameters and ingredients, including, for example, various types of controlled release compositions and coatings.Therefore, the compound is formulated with suitable excipients into a pharmaceutical composition that releases the compound in a controlled manner when administered.Examples include single or multiple unit tablet or capsule compositions, oil solutions, suspensions, emulsions, microcapsules, microspheres, molecular complexes, nanoparticles, patches, and liposomes.

[0186] In some embodiments, the compositions may comprise a "vectorized" form by encapsulation of the compound in a liposome or other encapsulation vehicle, or by immobilization of the compound, such as by covalent binding, chelation, or associative coordination, on a suitable biomolecule, such as one selected from proteins, lipoproteins, glycoproteins, and polysaccharides.

[0187] In some embodiments, the compositions may be incorporated into microspheres, microcapsules, nanoparticles, liposomes, etc. for controlled release. Additionally, the compositions may include suspending, solubilizing, stabilizing, pH adjusting agents, tonicity adjusting agents, and / or dispersing agents. Alternatively, the compounds may be incorporated into biocompatible carriers, implants, or infusion devices.

[0188] Materials for use in the preparation of microspheres and / or microcapsules are, for example, biodegradable / bioerodible polymers such as polyglactin, poly(isobutylcyanoacrylate), poly(2-hydroxyethyl-L-glutamine) and poly(lactic acid). Biocompatible carriers that can be used when formulating controlled release parenteral preparations are carbohydrates (e.g., dextran), proteins (e.g., albumin), lipoproteins, or antibodies. Materials for use in implants can be non-biodegradable (e.g., polydimethylsiloxane) or biodegradable (e.g., poly(caprolactone)), poly(lactic acid), poly(glycolic acid), or poly(orthoesters), or combinations thereof.

[0189] In all embodiments, the compounds or other active compounds may be present as pharma- ceutically acceptable salts or other derivatives of the active compounds, such as ether derivatives, ester derivatives, acid derivatives, and derivatives that modify water solubility. Derivatives include individual enantiomers, diastereomers, racemates, and other isomers of the compounds. Derivatives also include all polymorphs and solvates of the compounds, such as hydrates, and those formed with organic solvents. Such isomers, polymorphs, and solvates may be prepared by methods known in the art, such as regiospecific and / or enantioselective synthesis and separation.

[0190] The ability to prepare salt depends on the acidity or basicity of the compound.Suitable salts of the compound include, but are not limited to, the salts made with acid addition, such as hydrochloric acid, hydrobromic acid, hydroiodic acid, percolating acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid, benzoic acid, carbonic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, hydroxyethanesulfonic acid, benzoic acid, p-toluenesulfonic acid, cyclohexanesulfamic acid, salicylic acid, p-aminosalicylic acid, 2-phenoxybenzoic acid, and 2-acetoxybenzoic acid; the salts made with saccharin; alkali metal salts, such as sodium and potassium salts; alkaline earth metal salts, such as calcium and magnesium salts; and the salts made with organic or inorganic ligands, such as quaternary ammonium salts.

[0191] Additional suitable salts include, but are not limited to, acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycolylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthalate, iodide, isothiocyanate, of the compound. Includes salts, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methyl bromide, methyl nitrate, methyl sulfate, mucoate, napsylate, nitrate, N-methylglucamine ammonium salt, oleate, pamoate (embonate), palmitate, pantothenate, phosphate / diphosphate, polygalacturonate, salicylate, stearate, sulfate, subacetate, succinate, tannate, tartrate, theoclate, tosylate, triethiodide and valerate.

[0192] Pharmaceutically acceptable acid addition salts can also exist as various solvates, for example with water, methanol, ethanol, dimethylformamide, etc. Mixtures of such solvates can also be prepared. The source of such solvates may be derived from the solvent of crystallization, from intrinsic solvents in the solvent of preparation or crystallization, or from exogenous solvents to such solvents.

[0193] Wetting agents, emulsifying agents, and lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring, and perfuming agents, preservatives, and antioxidants may also be present in the composition.

[0194] Examples of pharma- ceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylhydroxytoluene (BHT), lecithin, propyl gallate, alpha tocopherol, and the like; and (3) metal chelators, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0195] Unless the context clearly indicates otherwise, the compositions of all embodiments may include various pharma- ceutically acceptable salts, or other derivatives of the above.

[0196] The formulation and preparation of such compositions are well known to those skilled in the art of pharmaceutical formulation. Formulations can be found in Remington: The Science and Practice of Pharmacy.

[0197] The amount of compound employed for use in the present disclosure will vary depending on the condition, the patient / subject, and the severity of the condition.

[0198] The contents of all references (including literature, issued patents, and published patent applications) cited throughout this application are hereby expressly incorporated herein by reference. The present disclosure and methods and methods of making and using them are described in such full, clear, concise and exact terms as to enable any person skilled in the art to which it pertains, to make and use the same.

[0199] The actual dosage level of the active ingredient in the pharmaceutical composition may be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition and method of administration without causing toxicity to the patient.

[0200] The selected dosage level will depend upon a variety of factors, including the activity of the particular compound or combination of compounds or their esters, salts or amides employed, the route of administration, the time of administration, the rate of excretion of the particular compound(s) employed, the duration of treatment, other drugs, compounds and / or materials used in combination with the particular compound(s) employed, the age, sex, weight, medical condition, general health and prior medical history of the patient being treated, and similar factors well known in the medical arts.

[0201] A physician or veterinarian skilled in the art can easily determine and prescribe the therapeutically effective amount of the pharmaceutical composition required. For example, a physician or veterinarian will start with a dosage level of the pharmaceutical composition or compound that is lower than the amount required to obtain the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved. The term "therapeutically effective amount" refers to the concentration of the compound sufficient to induce the desired therapeutic effect. It is generally understood that the effective amount of the compound varies according to the subject's weight, sex, age, and medical history. Other factors that affect the effective amount may include, but are not limited to, the severity of the patient's condition, the disorder being treated, the stability of the compound, and other types of therapeutic agents that are administered together with the compound of the present disclosure, if necessary. Even larger total doses can be delivered by multiple administrations of the drug. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13 ed., 1814-1882; which is incorporated herein by reference).

[0202] Generally, a suitable daily amount of an active compound used in the compositions and methods of the present disclosure will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective amount will generally depend upon the factors described above.

[0203] The dosage may be a single dose or a cumulative dose (sequential administration), and can be easily determined by one skilled in the art. For example, the treatment may include only one administration of an effective amount of the pharmaceutical composition disclosed herein. Alternatively, the treatment may include multiple administrations of an effective amount of the pharmaceutical composition administered over a range of time periods, such as once a day, twice a day, three times a day, once every few days, or once a week. The timing of administration may vary from individual to individual, depending on factors such as the severity of the individual's symptoms. For example, an effective amount of the pharmaceutical composition disclosed herein may be administered to an individual once a day for an indefinite period of time, or until the individual no longer requires treatment. One skilled in the art will recognize that the individual's condition will be monitored throughout the course of treatment, and the effective amount of the pharmaceutical composition disclosed herein administered can be adjusted accordingly.

[0204] If necessary, the effective daily amount of active compound can be administered as 1, 2, 3, 4, 5, 6 or more divided doses, optionally in unit dosage form, administered separately at appropriate intervals throughout the day.In certain embodiments of the present disclosure, active compound can be administered two or three times a day.In a preferred embodiment, active compound is administered once a day.

[0205] In certain embodiments, the duration of administration of the therapeutic compound is 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or more. In certain embodiments, a treatment regimen may include a period during which administration is suspended for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or more.

[0206] The patient receiving this treatment will typically be any animal in need thereof, including primates, particularly humans and other mammals such as horses, cattle, pigs and sheep, as well as poultry and pets.

[0207] In other embodiments, the compounds described herein may be provided in a kit, either together in a single container or in separate containers, with one or more additional therapeutic agents, e.g., as separate pharmaceutical formulations that can be used together in a contemporaneous therapy as discussed herein. In certain such embodiments, the kit may further include instructions for co-administration of the pharmaceutical formulations, e.g., to treat or prevent any of the above-mentioned conditions.

[0208] Such combination preparations may employ the compounds of the present disclosure within the dosage ranges set forth hereinabove, or a pharma- ceutically acceptable salt thereof, and other pharma- ceutical active agents within their approved dosage ranges.

[0209] In some embodiments, the compound may be administered after a predicate event, such as after exposure to ionizing radiation or after the initiation of radiation exposure, including accidental or therapeutic radiation. In one embodiment, the compound is administered immediately after exposure. In another embodiment, the compound is administered within 12 hours of exposure. In another embodiment, the compound is administered within 24 hours of exposure. In another embodiment, the compound is administered 24 hours after exposure. In another embodiment, the compound is administered 24 hours after exposure. In another embodiment, the compound is administered 36 hours after exposure. In another embodiment, the compound is administered within 48 hours of exposure. In another embodiment, the compound is administered within 60 hours of exposure. In another embodiment, the compound is administered within 72 hours of exposure. In another embodiment, the compound is administered within 84 hours of exposure.

[0210] In certain embodiments or formulations, YEL002 / BCN057 was solubilized in aqueous solutions at physiologically compatible pH using 100 mM methanesulfonic acid (MSA) / 10% povidone (PVP); 100 mM MSA / 2% benzyl alcohol / 2% N-methylpyrrolidone (NMP); and 100 mM MSA / 10% ethanol / 1% poloxamer 188. In further aspects, 100 mM lactic acid was added to also improve the solubility of these mixtures. In yet another embodiment, a formulation containing YEL002 and 30% Captisol (SBE-β-CD) and 100 mM MSA provided excellent solubility at or above pH 4.1.

[0211] In another embodiment, formulations for intravenous, subcutaneous and oral delivery of therapeutic levels of YEL002 / BCN057 were developed containing 30% by weight Captisol (SBE-β-CD) and 100 mM MSA at pH 4.1 or higher (adjusted with 1.0 N NaOH).

[0212] Having now generally described the present disclosure, it will be more readily understood by reference to the following examples, which are included solely for the purpose of illustrating certain aspects and embodiments of the present invention and are not intended to limit the invention.

[0213] In other aspects of this embodiment, the molecules disclosed herein reduce the severity of the disease, e.g., by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%. In still other aspects of this embodiment, the fusion protein or chimeric molecule disclosed herein is, for example, about 5% to about 100%, about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 90%, about 30% to about reduce the severity of the disease by 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70%.

[0214] Finally, although aspects of the present specification are emphasized by referring to specific embodiments, it should be understood that those skilled in the art will readily recognize that these disclosed embodiments are merely illustrative of the principles of the subject matter disclosed herein. Accordingly, it should be understood that the disclosed subject matter is in no way limited to the specific compounds, compositions, articles, devices, methods, protocols, and / or reagents, etc. described herein, unless otherwise specified. In addition, those skilled in the art will appreciate that certain changes, modifications, permutations, variations, additions, cancellations, and subcombinations thereof can be made in accordance with the teachings of the present specification without departing from the spirit of the present specification. Accordingly, it is intended that the following appended claims and the claims hereafter introduced be construed to embrace all such changes, modifications, permutations, variations, additions, cancellations, and subcombinations as being within the true spirit and scope of those claims.

[0215] Certain embodiments of the present disclosure are described herein, including the best mode known to the inventors for carrying out the disclosure. Of course, variations on these described embodiments will become apparent to those skilled in the art upon reading the foregoing description. The inventors anticipate that those skilled in the art will employ such variations as necessary, and intend for the invention to be carried out in ways other than as specifically described herein. Accordingly, the present invention encompasses all modifications and equivalents of the subject matter recited in the appended claims as permitted by applicable law. Moreover, any combination of the above-described embodiments in all possible variations thereof is also encompassed by the present disclosure, unless otherwise indicated herein or otherwise clearly contradicted by the context.

[0216] Grouping of alternative embodiments, elements, or steps of the invention should not be construed as limiting the disclosure. Each group member may be referenced and claimed individually or in any combination with other group members disclosed herein. It is anticipated that one or more members of a group may be included in, or removed from, a group for reasons of convenience and / or patentability. When any such inclusion or deletion occurs, the specification shall be deemed to include the group as modified and thus satisfying the recitation requirements of all Markush groups used in the appended claims.

[0217] All patents, patent publications, and other publications mentioned or identified in this specification are individually and expressly incorporated herein by reference in their entirety, for example, to describe and disclose the compositions and methods described in such publications and that can be used in connection with the present disclosure. These publications are provided solely for their disclosure prior to the filing date of this application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by reason of prior disclosure or for any other reason. Any dates or representations as to the contents of these documents are based on the information available to the applicants and do not constitute any admission as to the accuracy of the dates or contents of these documents.

[0218] Finally, the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present disclosure, which is defined only by the claims. Thus, the present disclosure is not limited to the exact same that has been shown and described.

[0219] An embodiment is a compound of formula I: [ka] The compound includes R 1 = alkyl, alkylamine, ether, aryl or aryl halide, R2 =H or CH3, R 3 =H, OH or OCH3.

[0220] An embodiment is a compound of formula I: [ka] The compound includes R 1 = [ka] where n=1, 2 or 3; X=F, CF3Cl, OH or CH2OH; R 2 =H or CH3, R 3 =H, OH or OCH3.

[0221] An embodiment is a compound of formula I: [ka] The compound includes R 1 = [ka] and n=1, 2 or 3; X=F, CF3Cl, OH or CH2OH; Y=O, S or N, and R 2 =H or CH3, R 3 =H, OH or OCH3.

[0222] An embodiment is a compound of formula I: [ka] The compound includes R 1 = [ka] and n=1, 2 or 3; X=F, CF3Cl, OH or CH2OH; Y=O, S or N; R 2 =H or CH3, R 3 =H, OH or OCH3.

[0223] An embodiment is a compound of formula I: [ka] The compound includes R 1 = [ka] and n=1, 2 or 3, and R 2 =H or CH3, R 3 =H, OH or OCH3.

[0224] An embodiment is a compound of formula I: [ka] The compound includes R 1 = [ka] and n=1, 2 or 3, and R 2 =H or CH3, R 3 =H, OH or OCH3.

[0225] An embodiment is a compound of formula I: [ka] The compound includes R 1 = [ka] and n=1, 2 or 3, and R 2 =H or CH3, R 3 =H, OH or OCH3.

[0226] An embodiment is a compound of formula A: [ka] R 1 =H or CH3, R 2 is one of the following: [ka]

[0227] In some embodiments, one or more compounds described herein may be used therapeutically to treat cancer, as an immune checkpoint regulator, modulate PD-1 expression, enhance T cell responses or increase immune cell activity to treat disease, ameliorate one or more side effects of chemotherapy or radiation therapy, prevent or treat radiation-induced damage to epithelial cells, treat infectious diseases such as viral infections, and / or reduce inflammation. [Table 2] TIFF2024533229000034.tif228159TIFF2024533229000035.tif240159TIFF2024533229000036.tif239159 TIFF2024533229000037.tif239159TIFF2024533229000038.tif223159TIFF2024533229000039.tif159159

Claims

1. Formula I: 【Chemical 1】 or an analog thereof, wherein R ２ = H or CH ３ and R ３ = H, OH, OCH ３ , OCH2CH3, or CH2OCH3; R 1 = alkyl, alkylamine, ether, aryl, aryl halide, hydrogen, amino, amide, F, Cl, Br, I, nitro, alkoxy, hydroxyl, thiol, alkylthio, acylcarboxylic acid, ester, sulfonyl, sulfonamide, -SO 4 H, optionally substituted C1-C20 alkyl, optionally substituted C1-C20 alkenyl, optionally substituted C1-C20 alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted heterocyclic alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl halide, optionally substituted phenyl, or 【Chemistry 2】 【change】 is a functional group selected from n=1, 2, or 3; X = F, CF3Cl, OH or CH2OH; Y=O, S or N; Z = CH3, CH2OH, OCH, OCH2CH3, or CH2OCH3; Compound or its analogue.

2. 10. A composition for use in treating cancer in a subject, comprising a compound of claim 1.

3. 3. The composition of claim 2, wherein the cancer is bladder cancer, brain cancer, breast cancer, colorectal cancer, cervical cancer, gastrointestinal cancer, genitourinary cancer, head and neck cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, rectal cancer, skin cancer, blood cancer, testicular cancer, biliary tract cancer, bladder cancer, ganglionic cancer (e.g., neuroblastoma), leukemia, lymphoma, liver cancer (e.g., hepatocellular carcinoma), lung cancer (e.g., large cell carcinoma, non-small cell carcinoma, and squamous cell carcinoma), soft tissue cancer (e.g., angiosarcoma, leiomyosarcoma, liposarcoma, rhabdomyosarcoma, myxoma, and malignant fibrous histiocytoma), gastric cancer, or thyroid cancer.

4. The composition of claim 2, wherein the subject is administered one or more additional therapeutic, chemotherapeutic or radiotherapeutic agents.

5. The composition of claim 4, wherein the one or more additional therapeutic agents are a small molecule, an antibody, an antibody fragment, an antibody conjugate or an immunomodulator.

6. A composition for use in treating one or more side effects of chemotherapy or radiation therapy in a subject, the composition comprising a compound described in claim 1.

7. A composition for use in preventing or treating radiation-induced damage to epithelial cells in a subject, comprising the compound of claim 1, wherein the radiation-induced damage to the epithelial cells is identified as one or more of radiation-induced gastrointestinal syndrome (RIGS), radiation-induced mucositis, radiation-induced oral mucositis, radiation-induced proctitis, and radiation-induced enteritis.

8. A composition for use in treating a disease in a subject, comprising a compound of claim 1 or a compound of Formula II, Formula III, or Formula IV, wherein Formula II, Formula III, and Formula IV are defined as follows: 【Chemistry 3】 (In the formula, R 1 is alkyl, alkylamine, ether or selected from the group consisting of hydrogen, amino, amido, F, Cl, Br, I, nitro, alkoxy, hydroxyl, thiol, alkylthio, acylcarboxylic acid, ester, sulfonyl, sulfonamide, —SO 4 H, optionally substituted C1-C20 alkyl, optionally substituted C1-C20 alkenyl, optionally substituted C1-C20 alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted heterocyclic alkyl, optionally substituted heteroaryl, or optionally substituted phenyl; R 3 is H, OH, OCH 3 , OCH 2 CH 3 , or CH 2 OCH 3 ; R 2 , R 4 , and R 5 are H or CH 3 ; 【Chemistry 4】 (In the formula, R 3 is O—CH 3 ; R 2 is CH 3 or H; R 1 is 【Chemistry 5】 (one of the following); 【Chemistry 6】 (In the formula, R 3 is 【Chemistry 7】 wherein Y is OCH2CH3, OCH(n)CH3, NH2, OH, OCH3, CH3, H, CH2OH, BH2, SeCH3 or SCH3; R 1 and R 2 are independently hydrogen; a substituted or unsubstituted straight or branched C1-C20 alkyl, alkenyl, or alkynyl; a substituted or unsubstituted cycloalkyl, cycloalkenyl, heterocyclic alkyl, or heterocyclic alkenyl; a phenyl, substituted phenyl, aryl, substituted aryl, amino, amido, F, Cl, Br, I, nitro, hydroxyl, thiol, alkylthio, selenol, alkylselenyl, silyl, siloxy, boryl, carboxylic acid, sulfolyl, —SO 4 H, alkoxy, or acyl group.

9. 9. The composition of claim 8, wherein the disease is at least one of cerebral malaria, Trypanosoma cruzi-induced myocarditis, influenza virus A, Mycobacterium tuberculosis, chlamydia pulmonary infection, COPD, acute lung injury, liver infection (HBV or HCV), pancreatitis, type 1 diabetes, chronic infection, sepsis, autoimmune disease, or HIV.

10. The composition described in claim 9, wherein the autoimmune disease is at least one of Hashimoto's thyroiditis and Graves' disease (GD), rheumatoid arthritis (RA), systemic lupus erythematosus (SLE) or multiple sclerosis (MS).

11. The composition described in claim 8 for use in modulating PD-1 expression, enhancing T cell responses, or enhancing immune cell activity to treat a disease.

12. The composition of claim 8 for use in inhibiting PD-1, PD-L1 and / or PD-1 / PD-L1 interaction to treat a disease.

13. The composition described in claim 8 for use in blocking the interaction of PD-L1 with PD-1 and / or CD80 to treat a disease.

14. The composition of claim 11, 12 or 13, wherein one or more additional therapeutic agents are administered to the subject, the therapeutic agents being an immune checkpoint regulator, a small molecule, an antibody, an antibody fragment, an antibody conjugate or an immunomodulator, a chemotherapeutic agent or a radiotherapeutic agent.

15. The composition described in claim 8 for use in suppressing the expression of LAG-3 to treat a disease.

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