Compositions and methods for making and using small molecules for the treatment of health conditions

Abstract

The novel compounds, compositions, and methods disclosed herein are effective for treating subjects suspected of having or developing a condition, such as cancer (i.e., brain cancer or cancer that has metastasized to the brain). A method for treating cancer can include administering to a subject a pharmaceutical composition comprising at least one of the novel compounds disclosed herein, and optionally one or more additional anti-cancer therapies. The compounds, compositions, and methods can further be used for drug screening and other detection and isolation methods.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is a non-provisional patent application and claims the benefit of U.S. Provisional Patent Application No. 63 / 555,767, filed February 20, 2024, U.S. Provisional Patent Application No. 63 / 591,709, filed October 19, 2023, and U.S. Provisional Patent Application No. 63 / 448,964, filed February 28, 2023, the specifications of which are incorporated herein by reference in their entireties.

[0002] The present invention features compounds, compositions, and methods for making and using small molecules to treat health conditions. In some embodiments, the compositions include one or more novel compounds, tagged compounds, prodrugs, drug conjugates, and metabolites, as well as intermediate compounds thereof. In other embodiments, the present invention features compounds, compositions, and methods for treating and / or managing and / or preemptively preventing and / or reducing and / or significantly decreasing cancer, including, but not limited to, glioblastoma, cancers that metastasize to the brain, and other brain cancers. In other embodiments, the compounds, compositions, and methods relate to the treatment of cancer, including, but not limited to, one or more of brain cancer, breast cancer, skin cancer, metastatic cancer, pancreatic cancer, lung cancer, kidney cancer, liver cancer, bladder cancer, osteosarcoma, ovarian cancer, rectal cancer, blood cancer, gastrointestinal cancer, or any combination thereof. In further embodiments, the compounds, compositions, and methods relate to drug screening and other methods for generating novel compounds. [Background technology]

[0003] Despite advances in oncological treatments, cancer remains a leading cause of death due to its high morbidity and mortality. While small molecule therapies can successfully treat some cancers, many have narrow therapeutic indices, are not highly selective, and cause undesirable drug toxicity in patients. Insufficient penetration into sanctuary sites (e.g., the CNS) necessitates the administration of some small molecules at high concentrations, which can further contribute to toxic side effects. Furthermore, in many cancers, drug resistance to some small molecule therapies develops over time, leading to disease recurrence. Therefore, there is a need to create novel small molecules for more successful treatment of cancer and other health conditions. Summary of the Invention

[0004] The present invention features compounds, compositions, and methods that enable the production and use of small molecules for the treatment of health conditions. In some embodiments, the present disclosure provides novel compounds, compositions, and methods for treating one or more cancers that metastasize to the brain in a subject in need thereof. In other embodiments, the compounds, compositions, and methods relate to drug screening and other methods for generating novel compounds. Embodiments of the present invention are set forth in the dependent claims. If the embodiments of the present invention are not mutually exclusive, they may be freely combined with each other.

[0005] As discussed herein, the compounds, compositions and methods cross the blood-brain barrier (BBB) ​​and act as tubulin-targeted therapies specific to brain cancer, and are effective in significantly depolymerizing tubulin to stop cell division, kill cancer cells, and reduce tumor growth.

[0006] While not wishing to limit the present invention to any theory or mechanism, it is believed that the technical features of the present invention advantageously provide compounds and compositions with improved ability to cross the blood-brain barrier compared to other known products on the market, many of which fail to enter the brain and others whose success rate at crossing the BBB is much lower than that observed herein. For example, some molecules of the present application are designed to bind to small clefts on the surface of tubulin protein, allowing smaller molecules to enter, improving brain penetration and avoiding efflux pumps that would otherwise remove these molecules from the brain. Furthermore, it is believed that the present invention advantageously provides highly effective and potent treatments for cancer, including brain cancer (e.g., treatment-resistant glioblastoma) and cancers that metastasize to the brain, including, but not limited to, lung cancer, breast cancer, and melanoma. None of the currently known prior art documents possess the unique inventive technical features of the present invention.

[0007] Suitable methods and materials for practicing and / or testing embodiments of the present disclosure are described below. Such methods, materials, and examples are illustrative only and are not intended to be limiting. Other methods and materials similar or equivalent to those described herein can be used. For example, conventional methods well known in the art to which the present disclosure pertains are described in various general and more specific references.

[0008] In certain embodiments, the present disclosure provides compounds for use to treat, prevent, or ameliorate a health condition in a subject, or for use as a combination therapy in treating, reducing the occurrence of, or ameliorating a health condition in a subject in need thereof.

[0009] In some embodiments, the present invention provides a compound of formula (ID): [ka] The compound according to claim 1, In the formula, R5 is [ka] [ka] is selected from R6 is aryl or heteroaryl, and each aryl and heteroaryl may be independently unsubstituted or substituted with one or more alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfo, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, or heterocyclic alkylthio.

[0010] In other embodiments, R6 is one of the following: [ka]

[0011] In certain embodiments, the compounds are configured to cross the blood-brain barrier (BBB) ​​of a human or non-human subject. In certain embodiments, the compounds have a central nervous system multiparameter optimization (CNS MPO) score of 4.0 or greater, a Papp score of greater than 10, or an efflux ratio of the compound of less than 2.0.

[0012] In certain embodiments, the compounds are effective in treating a health condition, treating cancer or metastasis, preventing cancer cell division, inhibiting tubulin polymerization, destabilizing microtubules, arresting cell division in the G2 / M phase, being cytotoxic to multiple cancer cell lines, targeting blood vessels and vasculature in cancer or tumors, treating a non-neoplastic condition, treating gout, treating familial Mediterranean fever, treating onychomycosis, targeting blood vessels or vasculature, or any combination thereof.

[0013] In certain embodiments, the invention features a composition comprising a compound according to Formula (ID) and one or more of a tag, an inert moiety, or a targeting moiety linked to the compound. In some embodiments, the tag is a fluorescent tag, a radioactive tag, biotin, or any combination thereof. In some embodiments, the inert moiety is an ester, a carbamate, an aminoacyl ester, or any combination thereof. In other embodiments, the targeting moiety is an antibody, a polyethylene glycol (PEG) conjugate, or a long-chain polymer, a peptide sequence, or any combination thereof.

[0014] In some preferred embodiments, the compound is RGN6024: [ka] is.

[0015] In some embodiments, the present invention provides a compound of formula II-D: [ka] The compound according to claim 1, In the formula, J is [ka] [ka] and L is [ka] [ka] K is aryl or heteroaryl, each of which is independently unsubstituted or substituted with one or more groups selected from alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfo, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heterocyclyl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, and heterocyclic alkylthio.

[0016] In certain embodiments, the present invention is a compound further comprising K, wherein K is [ka] [ka] [ka] is selected from.

[0017] In certain embodiments, the compound is configured to cross the blood-brain barrier (BBB) ​​of a human or non-human subject. In other embodiments, the compound has a central nervous system multiparameter optimization (CNS MPO) score of 4.0 or greater, a Papp score of greater than 10, or an efflux ratio of the compound of less than 2.0.

[0018] In some embodiments, the invention features a composition including a compound according to formula (ID) disclosed herein. In other embodiments, the invention features a composition including a compound according to formula (II-D) disclosed herein.

[0019] In certain embodiments, the invention features a compound according to one of the following compounds or derivatives thereof: [ka] [ka] [ka] [ka]

[0020] In certain embodiments, the present invention is configured to cross the blood-brain barrier (BBB) ​​of a human or non-human subject, hi certain embodiments, the present invention has a central nervous system multiparameter optimization (CNS MPO) score of 4.0 or greater, a Papp score of greater than 10, or the compound has an efflux ratio of less than 2.0.

[0021] In some embodiments, the invention is a composition comprising any of the compounds disclosed herein.

[0022] Treatment method

[0023] In some embodiments, the invention features a method of preventing, delaying the onset of, or treating a health condition in a subject in need thereof, the method including: (a) identifying a subject exhibiting the health condition; and (b) administering to the subject a therapeutically effective amount of a composition comprising one or more compounds disclosed herein or derivatives thereof.

[0024] In certain embodiments, the present disclosure provides a method of treating a health condition in a subject who has, has had, is suspected of developing, or is at risk of developing the health condition, the method comprising administering to the subject a therapeutically effective amount of at least one compound disclosed herein.

[0025] In some embodiments, the subject is a human or non-human animal, hi other embodiments, the non-human subject is a livestock, companion animal, laboratory animal, or zoo animal, wild animal, reptile, fish, or bird.

[0026] In some embodiments, the health condition comprises one or more cancers. In certain embodiments, the health condition may be cancer, such as prostate cancer, brain cancer, breast cancer, skin cancer, metastatic cancer, pancreatic cancer, lung cancer, kidney cancer, liver cancer, bladder cancer, osteosarcoma, ovarian cancer, rectal cancer, blood cancer, gastrointestinal cancer, medulloblastoma, or other solid organ cancer, cellular or tissue cancer, or any combination thereof. In some embodiments, the cancer may be Ewing's sarcoma. In other embodiments, the cancer may be melanoma. In some embodiments, the health condition comprises brain cancer or a cancer that can metastasize to the brain. In some embodiments, the cancer is glioblastoma, high-grade glioma, other brain cancer, non-cell lung cancer (NSCLC) before or after metastasis to the brain, angiogenic cancer, or any combination thereof.

[0027] In some embodiments, the condition comprises a non-neoplastic condition. In some embodiments, the condition comprises gout, familial Mediterranean fever, or onychomycosis. In other embodiments, the condition comprises a vascular disease.

[0028] In some embodiments, the method further comprises administering one or more of an antimicrobial agent, a chemotherapeutic agent, other anti-cancer therapy, or an antibody or fragment thereof. In some embodiments, the antimicrobial agent comprises one or more of an antiviral, bactericidal, antifungal, or antibacterial agent, or other antimicrobial agent. In further embodiments, the antimicrobial agent can be an antibacterial agent (antibiotic) such as doxycycline, tetracycline, or other antibiotic, for example, a commonly applicable antibiotic.

[0029] In some embodiments, the chemotherapeutic agents comprise one or more of temozolomide, lomustine, velzutifan, cisplatin, carboplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin, doxorubicin, melphalan, roscovitine, mitomycin C, hydroxyurea, 5-fluorouracil, AraC (cytarabine), 6-mercaptopurine, 6-thioguanine, cisplatin, Ara-C, etoposide, gemcitabine, bortezomib, sunitinib, sorafenib, sodium valproate, HDAC inhibitors, DNA synthesis inhibitors, or dacarbazine, FLT3 inhibitors, farnesyltransferase inhibitors, topoisomerase II inhibitors, P-glycoprotein modulators, hypomethylating agents, or combinations thereof.

[0030] In some embodiments, the anti-cancer therapy comprises one or more of chemotherapy, radiation therapy, immunotherapy and / or surgery. In other embodiments, the anti-cancer therapy further comprises one or more anti-cancer therapies or treatments, such as one or more of chemotherapeutic agents, radiation therapy, small molecules, and immunomodulatory agents. In some embodiments, the anti-cancer therapy or treatment can be administered separately from the compounds disclosed herein or derivatives thereof.

[0031] In other embodiments, an antimicrobial agent, a chemotherapeutic agent, another anti-cancer therapy, or an antibody or fragment thereof is administered before, during, or after administration of one or more compounds or compositions disclosed herein or derivatives thereof.

[0032] In some embodiments, the condition is cancer and the one or more anti-cancer therapies comprise administration of one or more of temozolomide, lomustine, velzutifan, or any combination thereof before, during, or after administration of the composition. In other embodiments, the condition is NSCLC and the one or more anti-cancer therapies comprise administration of one or more of crizotinib, osimertinib, or any combination thereof before, during, or after administration of the composition.

[0033] In some embodiments, the composition is effective in preventing cancer cells from dividing. In some embodiments, the composition is effective in inhibiting tubulin polymerization. In other embodiments, the composition is effective in destabilizing microtubules.

[0034] In some embodiments, the therapeutically effective amount of the composition is determined based on the disorder being treated and the mechanism of delivery. In some embodiments, the therapeutically effective dose in humans is in the range of 0.5-5 mg / kg twice daily. In other embodiments, the therapeutically effective dose in mice is in the range of 1-25 mg / kg.

[0035] In some embodiments, the route of administration is one or more of intravenous injection, oral administration, subcutaneous injection, intramuscular injection, intrasternal injection, intrathecal administration, intratumoral, intravascular, intracerebral injection, intracisternal, intraventricular, intranasal or inhalation, parenteral, buccal, enteral, intraperitoneal, inhalable, infusion, intramuscular, ocular, intravitreal, otic, rectal, sublingual, topical, transdermal, intrapulmonary, intrauterine, vaginal, via ultrasound-induced blood-brain barrier disruption, implantable device, infusion technique, or nanoparticle-based delivery.

[0036] In some embodiments, the composition is in the form of a tablet, pill, coated tablet or coated pill. In some embodiments, the effective dose is administered to the subject before, during or after the administration of at least one compound at least once a day, every other day, every three days, twice a week, once a week, every other week, twice a month or once a month, every other month, every six months, or other suitable dosing regimen.

[0037] In some embodiments, the composition reduces the growth of drug-resistant glioblastoma (LN-18) tumors by 83% compared to placebo when administered orally daily at less than 25% of the toxic dose in a mouse model. In other embodiments, the compound reduces the growth of drug-resistant glioblastoma (LN-18) tumors by 86% compared to temozolomide (TMZ) when administered orally daily at less than 25% of the toxic dose in a mouse model.

[0038] In other embodiments, the method can further include administering to the subject a therapeutically effective amount of one or more anti-cancer agents or treatments before, simultaneously with, or after administering to the subject an effective amount of at least one compound or formulation disclosed herein. According to these embodiments, the one or more anti-cancer agent treatments can include radiation therapy. In some embodiments, the cancer can be brain cancer, and the one or more anti-cancer agents include administering one or more of temozolomide, lomustine, velzutifan, or any combination thereof. In other embodiments, the cancer is NSCLC, and the one or more anti-cancer treatments include administering to the subject one or more of crizotinib, osimertinib, or a combination thereof in addition to a compound or compound-containing formulation disclosed herein.

[0039] In certain embodiments, the compounds and compositions and their methods of use relate to RGN6024. According to these embodiments, the compounds and compositions can include, but are not limited to, RGN6024 and mixtures and pharmaceutical compositions thereof. In other embodiments, a method for treating, reducing the occurrence of, or preventing a condition in a subject can include administering a compound or composition or formulation comprising, but not limited to, RGN6024 in a pharmaceutically acceptable formulation. In certain embodiments, the condition is cancer. In some embodiments, the cancer is brain cancer. In some embodiments, a composition comprising, but not limited to, RGN6024 can be used in a formulation and administered to a subject in need thereof, optionally a brain cancer patient.

[0040] How to use

[0041] In some embodiments, the present invention relates to a method of modulating abnormal cell division, the method comprising the steps of (a) identifying a cell having abnormal cell division, and (b) administering a compound or composition disclosed herein or a derivative thereof.

[0042] In some embodiments, the present invention provides a drug screening method for identifying therapeutically effective drug candidates for treating a health condition in a subject in need thereof, the method comprising: (a) identifying an in vitro or in vivo model of the health condition; (b) administering a drug candidate and RGN6024 to the in vitro or in vivo model; (c) determining the efficacy, toxicity or side effects of the drug candidate and RGN6024; and (d) comparing the efficacy, toxicity or side effects of the drug candidate and RGN6024 to identify a therapeutically effective drug candidate.

[0043] In some embodiments, the present invention provides a method for detecting target cells having aberrant cell division, the method comprising: (a) identifying a sample containing target cells having aberrant cell division; (b) performing an affinity-based assay using a small molecule conjugated to a tag to contact the target cells having aberrant cell division with the small molecule conjugated to the tag; and (c) determining whether the small molecule conjugated to the tag binds to the target cells having aberrant cell division. In some embodiments, the small molecule is RGN6024, or a compound or composition disclosed herein or a derivative thereof. In further embodiments, the affinity-based assay is an immune-based assay, a receptor-based assay, an antibody-based assay, a nanoparticle-based assay, a chemical assay, an optical assay, or a kinetic binding assay. In other embodiments, the affinity-based assay is gel electrophoresis, an enzyme-linked immunosorbent assay, an immunoblot assay, a fluorescence intensity assay, a fluorescence anisotropy assay, a fluorescence energy transfer assay, a surface plasmon resonance (SPR) assay, a light scattering assay, a forward binding assay, a dissociation assay, or a reverse binding assay. In some embodiments, the affinity-based assay is a colchicine competitive binding assay.

[0044] In some embodiments, the present invention provides a method for detecting a protein expressed by a target cell having abnormal cell division, the method comprising: (a) identifying a sample containing a protein expressed by a target cell having abnormal cell division; (b) performing an affinity-based assay using a small molecule conjugated to a tag to contact the protein expressed by the target cell having abnormal cell division with the small molecule conjugated to the tag; and (c) determining whether the small molecule conjugated to the tag binds to the protein expressed by the target cell having abnormal cell division. In some embodiments, the small molecule is RGN6024, or a compound or composition disclosed herein or a derivative thereof. In further embodiments, the affinity-based assay is an immunoassay, a receptor-based assay, an antibody-based assay, a nanoparticle-based assay, a chemical assay, an optical assay, or a kinetic binding assay. In other embodiments, the affinity-based assay is gel electrophoresis, enzyme-linked immunosorbent assay, immunoblot assay, fluorescence intensity assay, fluorescence anisotropy assay, fluorescence energy transfer assay, surface plasmon resonance (SPR) assay, light scattering assay, forward binding assay, dissociation assay, or reverse binding assay. In some embodiments, the affinity-based assay is a colchicine competitive binding assay.

[0045] In some embodiments, the present invention provides a method for detecting or isolating target cells having aberrant cell division, the method comprising: (a) identifying a sample containing target cells having aberrant cell division; (b) performing an affinity-based pull-down assay using a small molecule conjugated to a tag to contact the target cells having aberrant cell division with the small molecule conjugated to the tag; and (c) selectively isolating the target cells having aberrant cell division. In some embodiments, the small molecule is RGN6024, or a compound or composition disclosed herein or a derivative thereof.

[0046] In another embodiment, a method for detecting or isolating a protein expressed by a target cell having aberrant cell division comprises: (a) identifying a sample containing a protein expressed by a target cell having aberrant cell division; (b) performing an affinity-based pull-down assay using a small molecule conjugated to a tag to contact the protein expressed by the target cell having aberrant cell division with the small molecule conjugated to the tag; and (c) selectively isolating the protein expressed by the target cell having aberrant cell division. In another embodiment, the small molecule is RGN6024, or a compound or composition disclosed herein or a derivative thereof.

[0047] kit

[0048] In some embodiments, the present invention is a kit for treating cancer, comprising: (a) a therapeutically effective dose of any one of the compositions disclosed herein or derivatives thereof; (b) at least one container for storing the composition; and (c) dosing and administration instructions.

[0049] In certain embodiments, the present disclosure provides kits for storing or storing any of the compounds and / or compositions, and kits for carrying out any of the methods disclosed herein. In some embodiments, the kits can include one or more compounds disclosed herein and / or one or more pharmaceutically acceptable formulations disclosed herein and at least one container. In some embodiments, the kits disclosed herein can be used to treat or prevent cancer in a subject.

[0050] Any feature or combination of features described herein is encompassed within the scope of the present invention, unless the features encompassed in any such combination are mutually inconsistent, as apparent from the context, the specification, and the knowledge of one skilled in the art. Additional advantages and aspects of the present invention are apparent in the following detailed description and claims.

[0051] The following drawings form part of the present specification and are included to further demonstrate certain embodiments of the present disclosure, which may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein. [Brief explanation of the drawings]

[0052] [Figure 1] FIG. 1 shows an exemplary experiment illustrating the chemical reaction scheme for synthesizing 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidin-5-yl)oxy)phenyl)cyclobutane-1-carboxamide (Compound B19) in accordance with certain embodiments of the present disclosure.

[0053] [Figure 2] FIG. 1 shows an exemplary experiment illustrating the chemical reaction scheme for synthesizing 3-methoxy-N-(4-((2-(4-methoxypiperidin-1-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)cyclobutane-1-carboxamide (compound B46), in accordance with certain embodiments of the present disclosure.

[0054] [Figure 3] FIG. 1 shows an exemplary experiment illustrating the chemical reaction scheme for synthesizing 3-methoxy-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide (compound B120) in accordance with certain embodiments of the present disclosure.

[0055] [Figure 4] FIG. 1 shows an exemplary experiment illustrating the chemical reaction scheme for the synthesis of 3-methoxy-N-(5-(4-morpholinophenoxy)thiazol-2-yl)cyclobutane-1-carboxamide (compound B121), according to certain embodiments of the present disclosure.

[0056] [Figure 5]FIG. 1 shows an exemplary experiment illustrating the chemical scheme for synthesizing N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxycyclobutane-1-carboxamide (compound B72) in accordance with certain embodiments of the present disclosure.

[0057] [Figure 6] FIG. 1 shows an exemplary experiment illustrating the chemical reaction scheme for synthesizing 3-methoxy-N-(3-methyl-4-((2-morpholinothiazol-5-yl)oxy)phenyl)cyclobutane-carboxamide (compound B71), in accordance with certain embodiments of the present disclosure.

[0058] [Figure 7] FIG. 1 shows an exemplary experiment illustrating the chemical scheme for synthesizing compound 126: 3-methoxy-N-(5-((6-morpholinopyridin-3-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide (compound B126), in accordance with certain embodiments of the present disclosure.

[0059] [Figure 8] FIG. 1 shows an exemplary experiment illustrating the chemical reaction scheme for synthesizing 3-methoxy-N-(5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide (compound B122) in accordance with certain embodiments of the present disclosure.

[0060] [Figure 9] FIG. 1 shows an exemplary experiment illustrating the chemical reaction scheme for synthesizing 3-methoxy-N-(5-(4-(piperidin-1-yl)phenoxy)thiazol-2-yl)cyclobutane-1-carboxamide (compound B125) in accordance with certain embodiments of the present disclosure.

[0061] [Figure 10]FIG. 1 shows an exemplary experiment illustrating the chemical reaction scheme for synthesizing N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxycyclobutane-1-carboxamide (compound B130), in accordance with certain embodiments of the present disclosure.

[0062] [Figure 11] FIG. 1 shows an exemplary experiment illustrating the chemical reaction scheme for synthesizing 3-methoxy-N-(5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide (compound B140), in accordance with certain embodiments of the present disclosure.

[0063] [Figure 12A] FIG. 1 shows an exemplary experiment illustrating the chemical reaction scheme for synthesizing 3-methoxy-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide (RGN6024), in accordance with certain embodiments of the present disclosure.

[0064] [Figure 12B] 1 shows the proposed metabolic pathways of RGN6024 in mouse, rat, dog, monkey, and human hepatocytes.

[0065] [Figure 13] FIG. 1 shows an exemplary experiment illustrating the chemical scheme for synthesizing N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide (compound B138) in accordance with certain embodiments of the present disclosure.

[0066] [Figure 14]FIG. 1 shows an exemplary experiment illustrating the chemical scheme for synthesizing N-(5-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide (compound B137), in accordance with certain embodiments of the present disclosure.

[0067] [Figure 15] FIG. 1 shows an exemplary experiment illustrating the chemical reaction scheme for synthesizing 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidin-5-yl)oxy)phenyl)bicyclo[1.1.1]pentane-1-carboxamide (compound B89), in accordance with certain embodiments of the present disclosure.

[0068] [Figure 16] FIG. 1 shows an exemplary experiment illustrating the chemical reaction scheme for synthesizing 3-methoxy-N-(5-(4-morpholinophenoxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide (compound B155), in accordance with certain embodiments of the present disclosure.

[0069] [Figure 17] FIG. 1 shows an exemplary experiment illustrating the chemical scheme for synthesizing N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide (compound B118) in accordance with certain embodiments of the present disclosure.

[0070] [Figure 18] FIG. 1 shows an exemplary experiment illustrating the chemical scheme for synthesizing N-(4-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide (compound B117) in accordance with certain embodiments of the present disclosure.

[0071] [Figure 19A]1 shows an exemplary experiment demonstrating the inhibition of tubulin polymerization. The effect of compounds B130, B144, and B147 at a concentration of 5 uM on tubulin polymerization is shown. Colchicine is a positive control.

[0072] [Figure 19B] 1 shows an exemplary experiment demonstrating the inhibition of tubulin polymerization. The effect of RGN6024 at a concentration of 5 uM on tubulin polymerization is shown. Colchicine (COL) is a positive control, and PX is paclitaxel.

[0073] [Figure 20A] 1 shows an exemplary experiment demonstrating the inhibition of tubulin polymerization. The effect of compound B138 at 0.1, 0.3, 1 and 3 uM concentrations is shown.

[0074] [Figure 20B] 1 shows an exemplary experiment demonstrating the inhibition of tubulin polymerization. The effect of RGN6024 at 0.3, 1, 3 and 5 uM concentrations is shown.

[0075] [Figure 21] 1 shows a fluorescence-based colchicine competitive binding assay using RGN6024 at a concentration of 50 uM, with nocodazole as a positive control, according to certain embodiments of the present disclosure.

[0076] [Figure 22] 1 shows an N,N'-ethylene-bis(iodoacetamide) (EBI) competition assay using compound B138 in MCF7 cell line, demonstrating binding of the compound at the colchicine binding site, according to certain embodiments of the present disclosure. Colchicine is a positive control, and vinblastine is a negative control. 3071 is an internal control compound.

[0077] [Figure 23] 1 shows an exemplary experiment showing the concentration of compound B138 in plasma after oral administration of 30 mg / kg to a rodent model, according to certain embodiments of the present disclosure.

[0078] [Figure 24] 1 shows an exemplary experiment showing the concentration of RGN6024 in plasma after oral administration of 30 mg / kg to a rodent model, according to certain embodiments of the present disclosure.

[0079] [Figure 25] 1 shows an exemplary experiment showing the concentration of RGN6024 in plasma and brain tissue after oral administration of 30 mg / kg to a rodent model, according to certain embodiments of the present disclosure.

[0080] [Figure 26A] 1 shows the amount of RGN6024 in plasma, brain, and tumor at the end of an efficacy study using a subcutaneous mouse model of the LN-18 cell line.

[0081] [Figure 26B] 1 shows the amount of Compound B155 in plasma, brain and tumor at the end of the efficacy study.

[0082] [Figure 27] Figure 27 shows tumors excised from an animal model and illustrates the effect of RGN6024 on tumor growth and size, comparing placebo and standard of care temozolomide (TMZ) with RGN6024.

[0083] [Figure 28] Figure 1 shows that RGN6024 reaches and penetrates glioblastoma tumors in mouse brain tissue. Mean concentrations of RGN6024 in plasma, brain, and tumor after oral administration of 7.5 mg / kg are shown. Samples were taken after the last dose of the tumor efficacy study.

[0084] [Figure 29]RGN6024 reduces tumor growth by 98% in an intracranial model of glioblastoma. BT-142 tumors (glioblastoma) were implanted into the brain. RGN6024 vs. placebo was administered orally at 15 mg / kg twice daily. The observed bioluminescent signal was roughly proportional to tumor size. Treated cells exhibited only 2% of the bioluminescent signal observed in untreated cells.

[0085] [Figure 30] Body weight as a proxy for health in an intracranial model of glioblastoma. Animals treated with RGN6024 maintained their weight compared to placebo-treated animals, which in contrast lost significant weight and showed increased tumor signal. The placebo group showed increased weight loss on days 18-21 compared to treatment.

[0086] [Figure 31] We show that RGN6024 induces G2 / M cell cycle arrest compared to control (vehicle only) and sabizablin (a clinical-stage tubulin-targeted therapy with minimal brain penetration). RGN6024 blocks brain cancer cells from dividing and arrests them in the G2 / M phase. Literature has shown that G2 / M phase cells are more sensitive to radiation therapy.

[0087] [Figure 32] The prospect of RGN6024 combination therapy is shown. Literature indicates that radiation therapy targets cancer cells during the G2 / M phase of cell division, which are more sensitive to radiation therapy. Preliminary results show an additive effect between radiation and RGN6024.

[0088] [Figure 33]We show that RGN6024 demonstrates dose-dependent inhibition of tubulin polymerization in biophysical assays. The dose-dependent effect of RGN6024 on the target indicates that (1) increasing doses result in increased inhibition of tubulin polymerization, consistent with binding to tubulin; and (2) RGN6024 (50 μM) achieves a similar level of tubulin polymerization inhibition compared to colchicine (positive control, 5 μM).

[0089] [Figure 34] Validation that RGN6024 binds to the colchicine binding site is shown. Colchicine binding within the colchicine pocket results in fluorescence. RGN6024 competes with colchicine for the binding pocket, as indicated by a decrease in fluorescence. This is consistent with RGN6024 targeting the colchicine binding site.

[0090] [Figure 35] Western blot target site engagement evidence in live cells is shown. RGN6024 blocks modification by binding at the colchicine binding site. Modified β-tubulin-N,N'-ethylene-bis(iodoacetamide) (EBI) selectively crosslinks β-tubulin at the colchicine binding site. GADPH (loading control). The EBI only lane (negative control) shows β-tubulin is modified in the absence of drug. The EBI + 6024 lane shows that RGN6024 reduces β-tubulin modification. The EBI + colchicine (positive control) lane shows β-tubulin is protected from modification.

[0091] [Figure 36] RGN6024 (5 μM concentration) is shown to be effective against 44 drug targets with the most severe adverse drug reactions, with inhibition of all targets below 30%. RGN6024 is suitable for preclinical testing.

[0092] [Figure 37]Shows RGN6024 efficacy in cancer cell lines (high-grade glioma cell lines: BT142, LN-18, U-87, LN-229, U118, and T98G; cell lines with a propensity for brain metastasis: SKMEL5, HCC1806, and NCI-H460).

[0093] [Figure 38] We show that RGN6024 is orally available in mice at well-tolerated doses. [Figure 38] Plasma concentrations (ng / mL) over time (8 hours) are shown for IV (3 mg / kg) vs. PO (30 mg / kg). Half-life (T1 / 2) (h) = 2.64. Peak concentration (CMAX) (ng / mL) = 7020. Oral bioavailability is greater than 50%, with a CMAX approaching approximately 17 μM.

[0094] [Figure 39] A 5-day maximum tolerated dose study is shown in which untreated mice were compared with mice treated with 10 mg / kg, 30 mg / kg, or 100 mg / kg of RGN6024 for 5 days. The oral maximum tolerated dose of RGN6024 is 30 mg / kg to 100 mg / kg. RGN6024 is water soluble in 30% 2-hydroxypropyl-β-cyclodextrin in saline.

[0095] [Figure 40] The physicochemical and ADME properties of RGN6024 are presented, including important brain penetration properties (molecular weight, LogD, MPO score), important oral administration properties (kinetic solubility), and cell permeability (MDR1-MDCK cells; Papp and efflux ratio). RGN6024 exhibits an MPO score >4.0 predicting good brain penetration, is water soluble in saline formulations, has significant permeability (Papp >10) leading to better diffusion, and has an efflux ratio <2.0, suggesting it is not an MDR1 pump substrate.

[0096] [Figure 41]The in vitro ADME properties of RGN6024 are shown. RGN6024 exhibits low clearance and a long half-life, indicating good plasma levels in humans, moderate protein binding in humans, a higher percentage of free drug than mouse data, and the potential for no significant inhibition of the cytochrome P450 isoforms tested.

[0097] [Figure 42] Demonstrating the greatest brain penetration of tubulin-targeted therapies, RGN6024 achieved 14-fold greater brain penetration compared to tubulin-targeted therapies ANG1005, sabizabulin, eribulin, paclitaxel, and unesubulin.

[0098] [Figure 43] RGN6024 plasma and brain profile in ICR mice following oral administration of 30 mg / kg in healthy (tumor-free) mice. Mean compound concentrations in plasma (ng / mL) and brain (ng / g) are compared over time (h). Brain Cmax=8.8 μM.

[0099] [Figure 44] We demonstrate that the in vitro efficacy of RGN6024 is comparable to other colchicine binding site therapies approved by the FDA or active in human clinical trials, and extends well beyond the realm of brain penetration.

[0100] [Figure 45] This study demonstrates the safety of FDA-approved and clinical-stage colchicine binding site therapies, including colchicine and sabizabulin. Although colchicine and sabizabulin have similar affinities for the tubulin protein, their safety profiles differ significantly.

[0101] [Figure 46] Side effect profiles of colchicine and sabizablin are shown. Figure 46 shows that despite having the same mechanism of action and target affinity, colchicine and sabizablin result in significantly different side effect profiles.

[0102] Figures 47A-C show the reversibility of RGN6024 binding to tubulin protein compared to colchicine (high toxicity) and sabizabrin (low toxicity), as evidenced by increased cell viability after washout. [Figure 47A] Figure 1 shows a small increase in cell viability after washout of colchicine. [Figure 47B] 1 shows an increase in cell viability after washout of sabizabrin. [Figure 47C] Figure 1 shows increased cell viability after washout of RGN6024. The reversibility of RGN6024's tubulin binding suggests improved safety in humans, similar to the reversibility of sabizabrin's tubulin binding, consistent with its low toxicity in humans.

[0103] [Figure 48] Figure 48 shows that tumor luminescence as a marker of tumor size. Figure 48 shows that RGN6024 reduces tumor size in an orthotopic mouse model using the human GBM cell line BT-142, using tumor luminescence as a marker of tumor size.

[0104] [Figure 49] Figure 49 shows the total weight of mice over several days after initiation of treatment with RGN6024. Figure 49 shows that vehicle (no drug) mice lose weight due to tumor burden in the brain. Treated mice (with RGN6024) lose less weight compared to vehicle mice.

[0105] [Figure 50] Figure 50 shows that RGN6024 inhibits tumor growth of LN-18 cells in mice. Figure 50 shows data for temozolomide (also known as TMZ, a standard of care drug) compared with RGN6024. Female CB17 SCID mice (n=8 / group) bearing LN-18 tumors were orally administered vehicle, temozolomide (TMZ, 25 mg / kg), and RGN6024 (7.5, 15, and 25 mg / kg) QD for 15 days.

[0106] [Figure 51] This shows that RGN6024 can be used to treat conditions such as cancer by targeting cell cycle elements and inducing cancer cell death. Figure 51 shows that RGN6024 targets and binds to tubulin. U-87 cells were plated at a density of 4000 cells / well in black 96-well plates (PhenoPlate, Perkin Elmer) and grown overnight. These cells were treated with compounds for 24 hours and then fixed with 4% paraformaldehyde for 20 minutes. Cells were washed with PBS and then permeabilized with FoxP3 perm buffer (BD Biosciences) for 10 minutes at room temperature. Cells were incubated with anti-TUBB3 (1:1000; Tuj1, STEMCELL Technologies) antibody overnight, followed by incubation with a secondary antibody for 1 hour. Nuclear DNA was labeled with Hoechst. Representative images (40x magnification) were obtained using an automated high-content imaging microscope (Operetta, Perkin Elmer).

[0107] [Figure 52] "Current FDA-approved therapies related to tubulin interactions bind to tubulin in large clefts on the protein's surface, are too large for efficient brain penetration, are substrates for efflux pumps, and therefore do not penetrate the brain. RGN6024 is smaller than the FDA-approved therapies, can passively cross the blood-brain barrier, and does not act as an efflux pump substrate."

[0108] [Figure 53] LN-18 xenograft tumors and brains from a mouse brain cancer model were excised at two time points after RGN6024 administration: 30 minutes for half of the mice and 90 minutes for the other half of the mice. The concentration of the target drug in the brain reflected the concentration of the target drug in the tumor at both time points. [Figure 55] 1 shows that brain concentrations of RGN6024 exceeded the amount required to shrink tumors in animals in 30 minutes.

[0109] [Figure 54] We demonstrate that RGN6024 was tested in a variety of high-grade glioma lines, as well as breast, lung, and melanoma tumors with a propensity for brain metastasis. A median efficacy of 91 nM was observed. In comparison, the brain penetration level of RGN6024 is 25-fold higher than the concentration required to see an effective response in these in vitro cancer models. DETAILED DESCRIPTION OF THE INVENTION

[0110] term

[0111] Unless otherwise explained, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosed invention belongs. The singular terms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Similarly, the word "or" is intended to include "and" unless the context clearly dictates otherwise.

[0112] The term "comprise" means that other elements may be present in addition to the defined elements presented. The use of "comprise" indicates inclusion rather than limitation. Stated another way, the term "comprise" means "including primarily, but not necessarily solely." Furthermore, variations of the word "comprise," such as "comprise" and "comprises," similarly have the same meaning. In one respect, the technology described herein relates to the compositions, systems, methods, and their respective component components described herein as essential to the invention, but still allows for the inclusion ("comprises") of unspecified elements, whether essential or not.

[0113] In order to facilitate review of the various embodiments of the disclosure, the following explanations of specific terms are provided:

[0114] As used herein, the term "about" can mean ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1% of a recited value, e.g., amount, dose, temperature, time, percentage, etc.

[0115] As used herein, the term "B137" or "compound B137" or "C21" or "compound C21" refers to a compound according to the following formula: N-(5-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide, having the structure: [ka] may be used interchangeably to refer to a compound having

[0116] As used herein, the term "B138" or "compound B138" or "C18" or "compound C18" refers to a compound according to the following formula: N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide, having the structure: [ka] may be used interchangeably to refer to a compound having

[0117] As used herein, the term "RGN6024" or "compound B139" or "B139" refers to a compound according to the following formula: 3-methoxy-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide, having the structure: [ka] may be used interchangeably to refer to a compound having

[0118] As used herein, the term "B141" or "compound B141" or "C2" or "compound C2" refers to a compound according to the formula: 3-methoxy-3-methyl-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide, having the structure: [ka] may be used interchangeably to refer to a compound having

[0119] As used herein, the term "B142" or "compound B142" or "C7" or "compound C7" refers to a compound according to the formula: 3-(cyclopropylmethoxy)-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide, having the structure: [ka] may be used interchangeably to refer to a compound having

[0120] As used herein, the term "B144" or "compound B144" or "C6" or "compound C6" refers to a compound according to the formula: 3-isopropoxy-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide, having the structure: [ka] may be used interchangeably to refer to a compound having

[0121] As used herein, the term "B146" or "Compound 146" or "C3" or "Compound C3" refers to a compound according to the formula: 3-methoxy-1-methyl-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide, having the structure: [ka] may be used interchangeably to refer to a compound having

[0122] As used herein, the term "B155" or "Compound 155" or "C27" or "Compound C27" or "C89" or "Compound C89" refers to a compound according to the formula: 3-methoxy-N-(5-(4-morpholinophenoxy)thiazol-2-yl)bicycle[1.1.1]pentane-1-carboxamide, having the structure: [ka] may be used interchangeably to refer to a compound having

[0123] As used herein, the term "B156" or "Compound 156" or "C14" or "Compound C14" refers to a compound according to the formula: 2-methoxy-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)cyclopropane-1-carboxamide, having the structure: [ka] may be used interchangeably to refer to a compound having

[0124] The term "analog," as used herein, refers to a chemical compound that is structurally similar to another compound (a so-called "reference" compound) but differs in composition, e.g., the substitution of one atom with an atom of a different element, or the presence of a particular functional group, or the substitution of one functional group with another functional group, or the absolute stereochemistry of one or more chiral centers of the reference compound. Thus, an analog is a compound that is similar or equivalent in function and appearance to the reference compound, but not in structure or origin.

[0125] As used herein, the term "isomers" refers to compounds that have the same number and types of atoms, and therefore the same molecular weight, but differ with respect to the structural arrangement or configuration of the atoms.

[0126] Unless otherwise stated, structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure, such as the R and S configurations of each asymmetric center, Z and E double bond isomers, and Z and E conformational isomers. Accordingly, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention. Unless otherwise stated, all tautomeric forms of the compounds of the invention are within the scope of the invention. Unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, replacement of hydrogen by deuterium or tritium, or 13 C-enriched carbon or 14 Compounds having this structure containing the replacement of a carbon with a C-enriched carbon are within the scope of the present invention. Such compounds are useful, for example, as analytical tools, probes in biological assays, or as therapeutic agents according to the present invention. In certain embodiments, the warhead portion of provided compounds contains one or more deuterium atoms.

[0127] As used herein, the term "ester" refers to a compound produced by modifying a functional group (e.g., a hydroxyl group, a carboxyl group, an amino group, etc.). Examples of "esters" include "esters formed with a hydroxyl group" and "esters formed with a carboxyl group." The term "ester" can refer to an ester whose ester residue is a "conventional protecting group" or a "protecting group removable in vivo by a biological method, such as hydrolysis." In some embodiments, the term "conventional protecting group" can refer to a protecting group that is removable by a chemical method, such as hydrogenolysis, hydrolysis, electrolysis, or photolysis. In other embodiments, the term "protecting group removable in vivo by a biological method, such as hydrolysis" can refer to a protecting group that is removable in vivo, such as by hydrolysis, to generate the free acid or a salt thereof after administration to a subject.

[0128] The terms "salts" and "pharmaceutically acceptable salts," as used herein, refer to salts that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, and allergic response, commensurate with a reasonable benefit-risk ratio, and are effective for their intended use. "Pharmacologically acceptable salts" can refer to salts that may be formed when the compounds herein possess acidic groups, such as carboxyl, or basic groups, such as amino or imino. In some embodiments, salts of the compounds disclosed herein can be formed with acidic groups and can include, but are not limited to, inorganic salts such as alkali metal salts, e.g., sodium, potassium, or lithium salts, alkaline earth metal salts, e.g., calcium or magnesium salts, metal salts, e.g., aluminum or iron salts, amine salts, e.g., ammonium salts, and organic salts such as t-octylamine, dibenzylamine, morpholine, glucosamine, phenylglycine alkyl ester salts, ethylenediamine, N-methylglucamine, guanidine, diethylamine, triethylamine, dicyclohexylamine, N,N'-dibenzylethylenediamine, chloroprocaine, procaine, diethanolamine, N-benzylphenethylamine, piperazine, tetramethylammonium, or tris(hydroxymethyl)aminomethane salts, as well as amino acid salts, e.g., glycine, lysine, arginine, ornithine, glutamate, aspartate.In some embodiments, salt derivatives of the compounds disclosed herein formed with basic groups may include, but are not limited to, hydrogen halides such as hydrofluoride, hydrochloride, hydrobromide or hydrogen iodide salts, inorganic acid salts such as nitrate, perchlorate, sulfate or phosphate salts, lower alkane sulfonates such as methanesulfonate, trifluoromethanesulfonate or ethanesulfonate, arylsulfonates such as benzenesulfonate or p-toluenesulfonate, organic acid salts such as acetate, malate, fumarate, succinate, citrate, ascorbate, tartrate, oxalate or maleate salts, amino acid salts such as glycine, lysine, arginine, histidine, ornithine, glutamate or aspartate salts. In certain embodiments, the pharmacologically acceptable salts of the compounds disclosed herein may absorb water to form hydrates for use in the formulations disclosed herein when exposed to air or recrystallized.

[0129] As used herein, the terms "active metabolite" and "metabolite" refer to biomolecules involved in metabolism. The term "metabolite" can also refer to intermediate or end products of metabolic reactions catalyzed by naturally occurring enzymes in cells. The term "metabolite" is used to describe small molecule compounds, but can also be defined as endogenous compounds such as amino acids, lipids, sugars, and organic acids. In addition to naturally occurring metabolites, metabolites can be artificially synthesized for industrial or pharmaceutical use.

[0130] As used herein, the terms "intermediate" and "intermediate compound" refer to a molecular entity (e.g., atom, ion, molecule, etc.) formed directly or indirectly from a reactant that is converted to a product in a multi-step chemical reaction. Reactive intermediates can be reactive, short-lived, and energetic, and typically react further to yield the final product.

[0131] As used herein, the term "tagged compound" refers to a compound that is tagged or labeled with a probe to aid in the detection and / or tracking of a biomolecule. Examples of tags may include, but are not limited to, biotin, fluorescent tags, radioisotopes, and hydrophobic tags.

[0132] The term "prodrug" as used herein refers to a compound that is made more active in vivo by the metabolism of a precursor drug.The compounds and compositions described herein can exist as prodrugs, for example, as described in Hydrolysis in Drug and Prodrug Metabolism: Chemistry, Biochemistry, and Enzymology (Testa, Bernard and Mayer, Joachim M. Wiley-VHCA, Zurich, Switzerland, 2003).The prodrugs described herein are structurally modified forms of compounds that easily undergo chemical changes under physiological conditions to produce active compounds.In addition, prodrugs can be converted into active compounds by chemical or biochemical methods in an ex vivo environment.

[0133] The term "drug conjugate" as used herein refers to the covalent attachment of a drug or prodrug to a natural or synthetic molecular carrier for a specific application. Drug conjugation can be used to control drug release, target drug delivery, improve drug stability (e.g., pharmacokinetics and pharmacodynamics), enhance drug solubility, and alter toxicity profiles. Drug conjugation can occur with polymers, proteins, antibodies, and the like. Examples of drug conjugates include small molecule drug conjugates (SMDCs), nanoparticles, antibodies, and peptide sequences. SMDCs enable targeted therapy and are composed of a low-molecular-weight, high-affinity targeting ligand, a linker, and a drug payload.

[0134] As used herein, the term "alkyl" refers to a C1-C 20It refers to saturated aliphatic hydrocarbon groups, including straight-chain and branched-chain groups. Representative examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylbutyl, Examples of alkyl groups include, but are not limited to, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and branched chain isomers thereof. The alkyl group may be a lower alkyl having 1 to 6 carbon atoms. Representative examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like.The alkyl group may be substituted or unsubstituted. If substituted, the substituents may have one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfo, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, heterocyclic alkylthio, carbonyl, carboxy, or carboxylic acid ester.

[0135] As used herein, the term "alkylsulfo" refers to an ester of an alkanesulfonic acid. As used herein, the term "alkylamino" refers to an alkyl substituent attached to an amino group. Representative examples include phenylalanino, threonine, tryptophan, tyrosine, valino, N 2 -Glutamino, N 2 -Histidino, N 4 -Asparagino, etc.

[0136] The term "cycloalkyl," as used herein, refers to a saturated and / or partially unsaturated monocyclic or polycyclic hydrocarbon group having 3 to 20 carbon atoms. Representative examples of monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like. Polycyclic cycloalkyls can include cycloalkyls having spirocyclic, fused, and bridged rings. Representative examples of polycyclic cycloalkyls include, but are not limited to, bicyclo[1.1.1]pentyl, bicyclo[2.1.1]hexyl, and bicyclo[2.2.2]octyl. The cycloalkyls herein can be substituted or unsubstituted. When substituted, the substituents may be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfo, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, heterocyclic alkylthio, carbonyl, carboxy, or carboxylic acid ester. As used herein, the term "cycloalkylthio" refers to a cycloalkyl ring bonded to a sulfur group. As used herein, the term "heterocyclic alkyl" refers to a cycloalkyl group derived from a cycloalkane by removing a hydrogen atom from a ring having atoms of at least two different elements as ring members. As used herein, the term "heterocyclic alkylthio" refers to a cycloalkyl group derived from a cycloalkane by removing a hydrogen atom from a ring having a sulfur atom as a ring member. The cycloalkyl, heterocyclic alkyl, and heterocyclic alkylthio groups herein may be substituted or unsubstituted.When substituted, the substituents can be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfo, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, heterocyclic alkylthio, carbonyl, carboxy, or carboxylic acid ester.

[0137] The term "alkoxyl," as used herein, refers to an alkyl group bonded solely to an oxygen atom. Representative examples of alkoxyl groups include, but are not limited to, methoxyl, ethoxyl, and the like. The term "cycloalkoxyl," as used herein, refers to a cycloalkyl group bonded to an oxygen atom. Representative examples of cycloalkoxyl groups include, but are not limited to, cyclomethoxyl, cycloethoxyl, and the like.

[0138] The term "heterocyclic alkoxyl" as used herein refers to a cycloalkoxyl group having atoms of at least two different elements as ring members.Cycloalkoxyl and heterocyclic alkoxyl herein can be substituted or unsubstituted.When substituted, the substituents can be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfo, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, heterocyclic alkylthio, carbonyl, carboxy or carboxylic acid ester.

[0139] The term "alkenyl" as used herein refers to an alkene, i.e., a fragment formed from a double bond, by removing one hydrogen atom from any carbon atom. Representative examples of alkenyl groups include, but are not limited to, allyl, isopropenyl, oleyl, phytyl, prenyl, vinyl, etc. The term "alkynyl" as used herein refers to an alkyne, i.e., a fragment formed from a triple bond, by removing one hydrogen atom from any carbon atom.

[0140] The term "aryl," as used herein, refers to an organic group derived from an aromatic ring in which one hydrogen atom has been removed from the ring. Representative examples of aryl groups include phenyl, naphthyl, tolyl, and xylyl. The term "heteroaryl," as used herein, refers to a 5- to 14-membered aryl having 1 to 4 heteroatoms selected from O, S, and N as ring atoms, with the remaining ring atoms being C. Examples of heteroaryl groups include furan, thiophene, pyridine, pyrrole, N-alkylpyrrole, pyrimidine, pyrazine, imidazole, and tetrazolyl. The heteroaryl herein can be fused to an aryl, heterocyclic alkyl, or cycloalkyl, and the ring connected to the parent structure is a heteroaryl. The heteroaryl herein can be substituted or unsubstituted. When substituted, the substituents can be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfo, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, heterocyclic alkylthio, carbonyl, carboxy, or carboxylic acid ester.

[0141] As used herein, the term "hydroxyl" refers to an -OH group. As used herein, "hydroxyalkyl" refers to -alkyl-OH, where alkyl is as defined above. As used herein, "halo" or "halogen" refers to fluoro, chloro, bromo, or iodo. As used herein, "thiol" refers to an organosulfur compound of the form R-SH, where R represents alkyl or other organic substituent. As used herein, "carbonyl" refers to -C(=O)-. As used herein, "nitro" refers to -NO2. As used herein, "cyano" refers to -CN. As used herein, "amino" refers to -NH2. As used herein, "carboxy" refers to -C(=O)OH. As used herein, "carboxylic acid ester" refers to -C(=O)O-alkyl.

[0142] The term "heterocyclyl" as used herein refers to a monovalent group formed by removing a hydrogen atom from any ring atom of a heterocyclic compound. Representative examples of heterocyclyl include non-aromatic monocyclic, bicyclic, tricyclic, or spirocyclic ring systems containing up to seven atoms in each ring. The heterocyclyl herein can be substituted or unsubstituted. If substituted, the substituents can be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfo, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, heterocyclic alkylthio, carbonyl, carboxy, or carboxylic acid ester.

[0143] As used herein, the terms "bicyclic" and "bicyclic ring" refer to a polycyclic molecule characterized by two connected rings with at least two common atoms. Examples of bicyclic rings include fused and bridged bicyclic rings. Non-limiting examples of fused bicyclic rings include, but are not limited to, bicyclo[4.3.0]nonane, bicyclo[3.3.0]octane, bicyclo[4.2.0]octane, and bicyclo[3.2.0]heptane. As used herein, the terms "bridged bicyclic" or "bridged bicyclic ring" refer to a molecule characterized by two rings joined together that share three or more atoms, with the two bridgeheads separated by a "bridge" containing at least one atom. Bicyclic compounds containing bridges are typically rigid structures with little flexibility. Non-limiting examples of bridged bicyclic rings include, but are not limited to, bicyclo[1.1.1]pentane, bicyclo[2.1.1]hexane, bicyclo[2.2.2]octane, and 5-oxaspiro[3.4]octane. The bicyclic rings herein can be substituted or unsubstituted. If substituted, the substituents can be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfo, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, heterocyclic alkylthio, carbonyl, carboxy, or carboxylic acid ester.

[0144] As used herein, the term "optionally substituted" indicates that a group may be unsubstituted or substituted with one or more substituents provided herein or known in the art. As used herein, "substituted" with respect to a group indicates that a hydrogen atom bonded to a member atom within the group has been replaced. It should be understood that the term "substituted" includes the provision that such substitution is in accordance with the allowed valences of the atom and substituents being substituted, and that the substitution results in a stable compound (e.g., one that does not spontaneously undergo transformation by rearrangement, cyclization, or elimination, etc.). In certain embodiments, a single atom can be substituted with two or more substituents, as long as such substitution is in accordance with the allowed valences of the atom. Suitable substituents are defined herein for each substituted or optionally substituted group.

[0145] As used herein, the term "health condition" refers to an illness, injury, disorder, or physical or mental condition.

[0146] The term "therapeutically effective amount" may refer to an amount sufficient to achieve the desired therapeutic result or to ameliorate undesired symptoms, but generally insufficient to cause adverse side effects. The specific therapeutically effective dose level for any particular patient will depend on various factors, including the disorder being treated and the severity of the disorder; the specific composition used; the patient's age, weight, overall health, sex, and diet; the time of administration; the route of administration; the excretion rate of the specific compound used; the duration of treatment; drugs used in combination with or concomitantly with the specific compound used, and similar factors well known in the medical field. For example, it is well within the skill of the art to start a dose of a compound at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dose until the desired effect is achieved. If desired, an effective daily dose can be divided into multiple doses for administration purposes. Consequently, a single-dose composition may contain such an amount, or a fraction thereof, that makes up a daily dose. The dose can be adjusted by the individual physician in the event of any contraindications. The dose may vary and can be administered in one or more doses daily, for one or several days, weekly, twice a week, etc. Guidance can be found in the literature on appropriate dosages for given classes of medications.

[0147] The exact amount of the composition required will vary from subject to subject, depending on the species, age, weight and general condition of the subject, the severity of the disorder being treated, the specific composition used, its mode of administration, etc. Therefore, it is not possible to specify an exact amount for every composition. However, the appropriate amount can be determined by those skilled in the art using only routine experimentation, taking into account the teachings of this specification.

[0148] In some examples, dosage can be administered to subject once a day or divided dosage throughout the day, according to the clinical response of subject to drug, which is determined by methods known in the art.This dosage can be administered to subject once a day, once a week or for several days, and then if subject responds quickly, can be stopped, or dosage can be administered every day until clinical response is observed.Those skilled in the art can monitor the clinical response of subject to administering composition, and administer additional dosage as necessary.It is contemplated that composition can be administered to subject every day, on alternating days, every week, or at any interval between.

[0149] In some embodiments, it is particularly advantageous to formulate the composition in dosage units for ease of administration and uniformity of dosage. Dosage units refer to physically discrete units suited as unitary doses for the subjects to be treated, each unit containing a predetermined amount of active compound calculated to produce the desired therapeutic effect.

[0150] Dosage can vary within this range depending on the dosage form used and the route of administration used.Dosages can be formulated in animal models to achieve a concentration range that includes the IC50 (i.e., the concentration of the test compound that achieves half-maximal suppression of symptoms) determined in cell culture.Such information can be used to more accurately determine useful doses in humans.

[0151] The term "pharmaceutically acceptable" can refer to any dosage form of a compound, formulation, or composition that is suitable for contact with human subjects or tissues, and, where appropriate, suitable for use in animals, and that is free of excessive toxicity, irritation, and reduced side effects or complications associated with its administration as a consumable or otherwise, commensurate with a reasonable benefit / risk ratio, within the scope of sound medical judgment.

[0152] As used herein, the terms "individual," "subject," "host," "animal," and "patient" can be used interchangeably and refer to any subject or any mammal, such as a human (e.g., adult, adolescent, infant, geriatric, child, baby, and fetus), companion animal (e.g., pet, horse), livestock, or other animal, for diagnosis, treatment, prevention, or therapy, as appropriate.

[0153] As used herein, the terms "treat," "treating," and "treatment" can refer to both therapeutic treatment and prophylactic or preventative measures aimed at preventing, reducing, slowing (alleviating), inhibiting, or eliminating an undesirable physiological change, symptom, disease, or disorder (e.g., cancer).

[0154] For purposes of the present invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, whether detectable or undetectable, reduction in the extent of disease, stabilization (i.e., not worsening) of disease, delay or slowing of disease progression, improvement or alleviation of disease state, and remission (whether partial or total). "Treatment" can also mean extending survival compared to the expected survival if not treated. Those in need of treatment include those already with a condition or disorder, as well as those prone to having a condition or disorder, or those in whom a condition or disorder is to be prevented or whose onset is to be delayed. Optionally, a subject or patient can be identified (e.g., diagnosed) as suffering from a disease or condition prior to administration of the compositions of the present invention. Subjects at risk for a disease or disorder can be identified, for example, by any one or combination of suitable diagnostic or prognostic assays known in the art.

[0155] As used herein, the term "administration" refers to the act of providing a drug, prodrug, or other agent or therapeutic treatment to a subject or to cells, tissues, and organs in vivo, in vitro, or ex vivo. Exemplary routes of administration to the human body can be through the subarachnoid space of the brain or spinal cord (subarachnoid space), the eye (ophthalmic), the mouth (oral), the skin (topical or transdermal), the nose (intranasal), the lungs (inhalation), the oral mucosa (buccal), the ear, the rectum, the vagina, injection (e.g., intravenous, subcutaneous, intratumoral, intraperitoneal, etc.), etc.

[0156] As used herein, the term "co-administration" or "co-administering" refers to the administration of two or more active ingredients simultaneously, immediately before, or immediately after the administration of one or more additional therapies. The compounds of the present disclosure may be administered alone or simultaneously with another compound or standard drug known in the art to a subject. Co-administration is meant to include simultaneous or sequential administration of the compounds individually or in combination. Dosage and administration intervals can be individually adjusted to provide an effective level of the administered compound for the specific clinical indication being treated. This provides a treatment regimen that is commensurate with the severity of the individual's disease state.

[0157] In one example, the compositions of the present invention can be administered orally, including but not limited to, powder or granules, suspensions or solutions in water or non-aqueous media, pills, lozenges, capsules, sachets or tablets.Thickeners, flavorings, diluents, emulsifiers, dispersing aids or binders may be desirable.Those skilled in the art who monitor the clinical response of the subject can adjust the administration frequency and dosage of the drug according to methods known in the art.

[0158] In another example, the compositions of the present invention may be administered intranasally or by inhalation. As used herein, "intranasal administration" refers to delivery of a composition to the nose and nasal cavity through one or both nostrils, and can include delivery by a spray mechanism (device) or a droplet mechanism (device), or via aerosolization of the composition, for example, by using a nasal spray, atomizer, dropper, or syringe. Administration of a composition by inhalation can be performed through the nose or mouth by a spray or droplet mechanism. As used herein, an "inhaler" can be a spray device or a droplet device for delivering a composition to the nasal cavity and upper and / or lower respiratory tract of a subject. Delivery can also be directly to any region of the respiratory system (e.g., lungs) via endotracheal intubation. A skilled artisan who monitors the subject's clinical response can adjust the administration frequency and dosage of the drug according to methods known in the art.

[0159] In another example, the compositions of the present invention can be administered by local intranasal administration (intranasal) or by inhalation. As used herein, "local intranasal administration" refers to delivery of a composition to the nose and nasal cavity through one or both nostrils and can include delivery by a spray or droplet mechanism or via aerosolization of the composition. Administration of a composition by inhalation can be through the nose or mouth by a spray or droplet mechanism. As used herein, an "inhaler" can be a spray or droplet device for delivering a composition in a pharmaceutically acceptable carrier to the nasal cavity and upper and / or lower respiratory tract of a subject. Delivery can also be directly to any region of the respiratory system (e.g., lungs) via endotracheal intubation. The exact amount of composition required will vary from subject to subject, depending on the species, age, weight, and general condition of the subject, the severity of the disorder being treated, the particular composition used, its mode of administration, etc. Therefore, it is not possible to specify an exact amount for every composition. However, an appropriate amount can be determined by one of ordinary skill in the art using only routine experimentation, given the teachings herein.

[0160] In one example, the compositions of the present invention can be administered by buccal or sublingual delivery. As used herein, "buccal delivery" can refer to an administration method in which the compound is delivered through the mucous membrane lining the cheek. In some embodiments, for buccal delivery, the composition is placed between the patient's gums and cheek. As used herein, "sublingual delivery" can refer to an administration method in which the composition is delivered through the mucous membrane under the tongue. In some embodiments, for sublingual delivery, the composition is administered under the patient's tongue.

[0161] In another example, the compositions of the present invention, if used, can be administered by parenteral administration of the composition, generally characterized by injection.Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for dissolving or suspending in liquid before injection, or as emulsions.A modified approach for parenteral administration includes the use of delayed-release or sustained-release systems to maintain a constant dosage.

[0162] In another example, the compositions of the present invention can be administered to a subject intramuscularly, for example, by using intramuscular injection or electroporation. One skilled in the art who monitors the subject's clinical response can adjust the administration frequency and dosage of the drug according to methods known in the art.

[0163] In one example, the compositions of the present invention can be administered via a lung lavage procedure. As used herein, "lung lavage" or "whole lung lavage" refers to a procedure in which a double-lumen endotracheal tube isolates one lung and injects sufficient saline or other pharmaceutically acceptable carrier into it to fill the entire volume of one lung. Furthermore, lung lavage can be used to deliver a composition in a pharmaceutically acceptable carrier to the lungs / lower respiratory tract of a subject. The lung undergoing lavage can be drained, repeatedly filled with fluid, and finally aspirated as many times as necessary. This procedure can be repeated with another lung at a different time. In some embodiments, administration of a composition via lung lavage can be used in patients who are extremely seriously ill and hospitalized in an intensive care unit (ICU).

[0164] In another example, the compositions of the invention can be contained in or stored in a container, bag, pack, or dispenser together with instructions for administration. For example, the instructions can include directions for administering the composition to a subject.

[0165] The following patents and applications are incorporated by reference herein in their entireties: U.S. Provisional Patent Application No. 63 / 555,767, filed February 20, 2024; U.S. Provisional Patent Application No. 63 / 591,709, filed October 19, 2023; U.S. Provisional Patent Application No. 63 / 448,964, filed February 28, 2023; and PCT / IB2022 / 062416, filed December 16, 2022.

[0166] All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety for all purposes. In case of conflict, the present specification, including explanations of terms, will control.

[0167] Briefly, the present invention features compounds and compositions for treating health conditions. In some embodiments, the compositions include one or more novel compounds, tagged compounds, prodrugs, drug conjugates, and metabolites, as well as intermediate compounds thereof. The compounds and compositions are clinically highly effective in crossing the blood-brain barrier (BBB) ​​and acting as tubulin-targeting therapies specific to brain cancer, significantly depolymerizing tubulin to halt cell division, kill cancer cells, and reduce tumor growth. While not wishing to limit the present invention to any theory or mechanism, it is believed that the technical features of the present invention advantageously provide highly effective and potent treatments for cancer, including brain cancer and cancers that metastasize to the brain, including, but not limited to, lung cancer, breast cancer, and melanoma. None of the currently known prior art documents possess the unique inventive technical features of the present invention.

[0168] The present invention also includes methods for treating and / or managing and / or preemptively preventing and / or reducing and / or significantly reducing cancer, including, but not limited to, glioblastoma and other brain cancers, comprising administering the compounds and compositions herein. In some embodiments, the compounds, compositions, and methods relate to the treatment of cancer, including, but not limited to, cancer that metastasizes to the brain. In other embodiments, the compounds, compositions, and methods relate to the treatment of cancer, including, but not limited to, one or more of brain cancer, breast cancer, skin cancer, metastatic cancer, pancreatic cancer, lung cancer, kidney cancer, liver cancer, bladder cancer, osteosarcoma, ovarian cancer, rectal cancer, blood cancer, gastrointestinal cancer, medulloblastoma, or any combination thereof.

[0169] The present invention further includes methods for modulating abnormal cell division.In other embodiments, the present invention relates to methods for isolating, detecting and screening novel drugs.

[0170] Referring now to Figures 1-54, the present invention features compounds, compositions, and methods for the production and use of small molecules for the treatment of health conditions, including, but not limited to, cancer. In some embodiments, the compositions include one or more novel compounds, tagged compounds, prodrugs, drug conjugates, and metabolites, as well as intermediate compounds thereof. The present invention also features compounds, compositions, and methods for treating and / or managing and / or preemptively preventing and / or reducing and / or significantly decreasing cancer, including, but not limited to, glioblastoma and other brain cancers. In further embodiments, the compounds, compositions, and methods relate to drug screening and other methods for the generation of novel compounds.

[0171] In the following sections, specific exemplary compositions and methods are described to detail specific embodiments of the present invention. It will be apparent to those skilled in the art that implementing a specific embodiment does not require the use of all or even some of the specific details outlined herein, but rather that concentrations, times, and other specific details can be varied through routine experimentation. In some cases, well-known methods or components are not included in the description.

[0172]

[0003] Embodiments disclosed herein relate to novel chemical compounds for use in treating or preventing a health condition. In certain embodiments, the chemical compounds disclosed herein can target microtubules. According to these embodiments, the chemical compounds disclosed herein can target and destabilize microtubules. Microtubule-destabilizing agents can be used to treat, prevent, or reduce the risk of developing a health condition. In certain embodiments, the health condition may include cancer or other health conditions.

[0173] Some embodiments disclosed herein relate to the disclosed compounds and the preparation of compositions containing at least these compounds disclosed herein, methods and processes of use.It is understood that each combination, subset, interaction, agent disclosed herein is contemplated, although specific reference to each individual combination and collective combination and permutation of these compounds cannot be explicitly disclosed.Further embodiments of the present disclosure are described below.

[0174] I. Compound

[0175] In certain embodiments, the present disclosure provides compounds for use to treat, prevent, or ameliorate a health condition in a subject, or for use as a combination therapy in treating, reducing the occurrence of, or ameliorating a health condition in a subject in need thereof.

[0176] In certain embodiments, the invention features compounds for use to treat, prevent, or ameliorate a health condition in a subject, or for use as a combination therapy in treating, reducing the occurrence of, or ameliorating a health condition in a subject in need thereof.

[0177] For example, in some embodiments, the present invention provides a compound of formula (IA): ((A m -B n ) j -C-[L'] q ) k -EL” s -G t (Formula IA) or an analog, isomer or pharmaceutically acceptable salt thereof, During the ceremony, A is H, alkyl, alkylcarbonyl, carboxyl, carboxylic ester, carboalkoxy, ester, heterocyclic alkyl, aryl, heteroaryl, haloalkyl, hydrocarbyl, alkenyl, alkynyl, phosphate, acetyl, or a combination thereof; m is 0 or 1, B is F, O or S, and when B is F, m=0; n is 0, 1 or 2; j is 0, 1, 2, or 3; C is a 3-, 4-, 5-, or 6-membered cyclic or bicyclic moiety optionally substituted by one or more heteroatoms or substituents; when n is 0, C is bonded to A; L' is at least one thioester, ester, -CONH-, or -SONH-, -NHCO-, or a linker comprising any combination thereof; q is 0, 1, 2, 3 or 4; k is 0, 1, 2, 3, or 4; E is a 3-, 4-, 5-, or 6-membered cyclic or bicyclic moiety optionally substituted by one or more heteroatoms or substituents; when q is 0, E is attached to C; L" is a linker, where L" is O, CH2, -S- or -C=O; s is 0 or 1, G is [ka] and t is 0 or 1, L" is attached to a carbon in one of the rings of G in place of any one of R17, R18, R19, R20, R21, R22 or R23; T, U and V are each CH or N; W is O, S, -CH(R24) or -CH(OR25), where R24 and R25 are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle, and substituted or unsubstituted cycloalkyl; R17, R18, R19, R20, R21, R22, and R23 are each independently selected from the group consisting of H, F, Cl, I, Br, straight-chain or branched-chain substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted heterocycle.

[0178] In some preferred embodiments, L" is attached to a carbon of G in place of any one of R17, R22, or R23.

[0179] In some embodiments, the present invention also provides a compound of formula (II-B): [ka] wherein J is any one of the following: [ka] [ka]

[0180] In certain embodiments, the compounds of the present disclosure can include any one of the compounds provided in Table 1.

[0181] [Table 1-1] [Table 1-2] [Table 1-3]

[0182] The present invention relates to a compound of formula (III-A): [ka] wherein K is one of the following: [ka] [ka]

[0183] In certain embodiments, the compounds of the present disclosure can include any one of the compounds provided in Table 2.

[0184] [Table 2-1] [Table 2-2]

[0185] The present invention relates to a compound of formula (IV-A): [ka] wherein K is one of the following: [ka] [ka]

[0186] In certain embodiments, the compounds of the present disclosure can include any one of the compounds provided in Table 3.

[0187] [Table 3-1] [Table 3-2]

[0188] The present invention also relates to a compound of formula (VA): [ka] wherein L is any one of the following: [ka] [ka]

[0189] In certain embodiments, the compounds of the present disclosure can include any one of the compounds provided in Table 4.

[0190] [Table 4-1] [Table 4-2] [Table 4-3] [Table 4-4] [Table 4-5]

[0191] The present invention also relates to a compound of formula (VI-A): [ka] wherein L is any one of the following: [ka] [ka]

[0192] In certain embodiments, the compounds of the present disclosure can include any one of the compounds provided in Table 5.

[0193] [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4] [Table 5-5]

[0194] The present invention relates to a compound of formula (VII-A): [ka] wherein L is any one of the following: [ka] [ka]

[0195] In certain embodiments, the compounds of the present disclosure can include any one of the compounds provided in Table 6.

[0196] [Table 6-1] [Table 6-2] [Table 6-3] [Table 6-4] [Table 6-5]

[0197] In certain embodiments, the invention features a compound according to one of the compounds provided in Table 7, or a derivative thereof.

[0198] [Table 7-1] [Table 7-2]

[0199] In some embodiments, the invention is a compound or composition comprising RGN6024 or a derivative thereof. [ka]

[0200] In some embodiments, the compound has formula IB: ABC-L'-EL”-G (Formula IB) and wherein: A is a straight or branched chain alkyl, cycloalkyl, or cycloalkylmethyl; B is O or S, and B is covalently bonded to any available atom on A; C is cycloalkyl or bicyclic alkyl and any available atom on C is covalently bonded to B; L' is a thioester, an ester, -CONH-, or -SONH-, -NHCO-, and any available atom on L' is covalently bonded to any available atom on C; E is thiazole and any available atom on E is covalently bonded to any available atom on L'; L" is a linker selected from the group consisting of O, CH2, -S-, and -C=O, and L" is covalently bonded to any available atom on E; G is [ka] wherein any available atom on G is covalently bonded to L″, L″ can be bonded to a carbon of a ring of G in place of any one of R17, R18, R19, R20, R21, R22, or R23, and any position not bonded to L″ remains designated; T and U are CH or N; V is CH or N; W is O, S, -CH(R24) or -CH(OR25), where R24 and R25 are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle, and substituted or unsubstituted cycloalkyl; R17, R18, R19, R20, R21, R22, and R23 are each independently selected from the group consisting of H, F, Cl, I, Br, straight-chain or branched-chain substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted heterocycle.

[0201] In certain embodiments, the compound of formula (IB) can include any one of the compounds provided in Table 8.

[0202] [Table 8-1] [Table 8-2]

[0203] In some embodiments, the compound can include a compound according to formula II-B or a derivative thereof: [ka] wherein X is selected from the group consisting of O, S, or N(R26), and R26 is selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; Z is selected from the group consisting of —CH or N; R1 is [ka] wherein Y is selected from the group consisting of O, S, —C═O, and a halide, and when Y is a halide, R13 is absent; R13 is selected from the group consisting of H, linear or branched, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted heterocycle; BB is a bridged bicyclic moiety; R14 and R15 are each independently selected from the group consisting of H, F, Cl, I, Br, linear or branched substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted heterocycle; R16 is -CO- or -SO2-, R2 is [ka] wherein O in formula II-B is covalently bonded to one carbon of the R2 ring in place of any one of R17, R18, R19, R20, R21, R22, or R23, and any position not bonded to R2 can remain designated; T and U are CH or N; V is CH or N; W is O, S, -CH(R24) or -CH(OR25), where R24 and R25 are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle, and substituted or unsubstituted cycloalkyl; Each of R17, R18, R19, R20, R21, R22 and R23 is independently selected from the group consisting of H, F, Cl, I, Br, straight or branched chain substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted heterocycle.

[0204] In certain embodiments, the compound of formula (II-B) can include any one of the compounds provided in Table 9.

[0205] [Table 9]

[0206] In some embodiments, the compound can include a compound according to formula III-B or a derivative thereof: [ka] During the ceremony, R5 is [ka] and R6 is [ka] and Z in formula III-B is covalently bonded to one carbon of the R6 ring in place of any one of R17, R18, R19, R20, R21, R22, or R23, and any position not bonded to R6 can remain designated; R11, R14, R15, R17, R18, R19, R20, R21, R22 and R23 are each independently selected from the group consisting of H, F, Cl, I, Br, linear or branched substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted heterocycle; R12 and R13 are each independently selected from the group consisting of H, linear or branched, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted heterocycle; M is selected from the group consisting of O, S, and N(R26), where R26 is selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; Q is selected from the group consisting of O, S and halide, and when Z is halide, R6 is absent; Y is selected from the group consisting of O, S, -C=O and a halide, and when Y is a halide, R13 is absent; BB is a bridged bicyclic moiety; W is selected from the group consisting of O, S, -CH(R24) and -CH(OR25), where R24 and R25 are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle, and substituted or unsubstituted cycloalkyl.

[0207] In certain embodiments, the compound of formula (III-B) can include any one of the compounds provided in Table 10.

[0208] [Table 10-1] [Table 10-2]

[0209] In some embodiments, the compound of the invention can be any one of the following: [ka]

[0210] In some embodiments, the compound has the formula (IC): [ka] and / or a compound according to the formula (I), its analogs, isomers, pharmaceutically acceptable salts and / or prodrugs, and / or formulations thereof; In the formula, R1 is [ka] [ka] is selected from R2 is alkyl, aryl, and heteroaryl, each of which is independently unsubstituted or substituted with one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfo, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, and heterocyclic alkylthio; R3 is selected from H, CH3 or F, and R4 is selected from H or F.

[0211] In some embodiments, R 1 of the compounds of formula (IC) disclosed herein is selected from the following: [ka] [ka]

[0212] In some embodiments, R 1 of the compounds of formula (IC) disclosed herein is selected from the following: [ka] [ka]

[0213] In some embodiments, R2 of the compounds of formula (IC) disclosed herein is selected from the following: -CH3, [ka] [ka]

[0214] In some embodiments, R2 of the compounds of formula (IC) disclosed herein is selected from the following: [ka] [ka]

[0215] In some embodiments, compounds of the present disclosure include compounds of formula (IC), analogs, isomers, pharmaceutically acceptable salts, and / or prodrugs thereof, wherein R is: [ka] [ka] is selected from R2 is -CH3, [ka] [ka] is selected from R3 is selected from H and CH3, and R4 is selected from H and F.

[0216] In certain embodiments, compounds of formula (IC) can have R1, R2, R3 and / or R4 as shown in Table 11. In other embodiments, compounds of the present disclosure can include any one of the compounds provided in Table 11.

[0217] [Table 11-1] Table 11-2 Table 11-3 Table 11-4 Table 11-5 Table 11-6 Table 11-7 Table 11-8 Table 11-9 Table 11-10 Table 11-11 Table 11-12 Table 11-13 Table 11-14 Table 11-15 [Table 11-16] [Table 11-17] [Table 11-18] [Table 11-19] [Table 11-20] [Table 11-21]

[0218] In some embodiments, provided compounds have formula (II-C): [ka] and / or a compound according to the formula: [ka] [ka] is selected from R6 is alkyl, aryl, and heteroaryl, each of which is independently unsubstituted or substituted with one or more of alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfo, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, or heterocyclic alkylthio.

[0219] In some embodiments, R5 is selected from the following: [ka] [ka]

[0220] In some embodiments, R5 is selected from the following: [ka] [ka]

[0221] In some embodiments, R5 is selected from the following: [ka] [ka]

[0222] In some embodiments, R6 is selected from the following: -CH3, [ka] [ka]

[0223] In some embodiments, R6 is selected from the following: [ka] [ka]

[0224] In some embodiments, the compounds of the present disclosure can be compounds of formula (II-C), analogs, isomers, pharmaceutically acceptable salts, and / or prodrugs thereof, and / or formulations thereof, wherein R5 is: [ka] [ka] is selected from R6 is -CH3, [ka] [ka] is selected from.

[0225] In certain embodiments, compounds of formula (II-C) can have R5 and / or R6 as shown in Table 12. In other embodiments, compounds of the present disclosure can include any one of the compounds provided in Table 12.

[0226] [Table 12-1] [Table 12-2] [Table 12-3] [Table 12-4]

[0227] In some embodiments, the compound has formula (III-C): [ka] and the compounds according to the formula (I), their analogs, isomers, pharmaceutically acceptable salts and / or prodrugs, In the formula, R7 is [ka] [ka] is selected from R8 is alkyl, aryl, and heteroaryl, each of which is independently unsubstituted or substituted with one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfo, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, and heterocyclic alkylthio; R9 is selected from H, CH3 or F, and R 10 is selected from H or F.

[0228] In some embodiments, R7 of the compounds of Formula (III-C) disclosed herein is selected from the following: [ka] [ka]

[0229] In some embodiments, R7 of the compounds of formula (IC) disclosed herein is selected from the following: [ka] [ka]

[0230] In some embodiments, R8 of the compounds of Formula (III-C) disclosed herein is selected from the following: -CH3, [ka] [ka]

[0231] In some embodiments, R8 of the compounds of Formula (III-C) disclosed herein is selected from the following: -CH3, [ka] [ka]

[0232] In some embodiments, R8 of the compounds of Formula (III-C) disclosed herein is selected from the following: [ka] [ka]

[0233] In some embodiments, R8 of the compound of Formula (III-C) is [ka] is.

[0234] In some embodiments, compounds of the present disclosure can be compounds of formula (III-C), analogs, isomers, pharmaceutically acceptable salts, and / or prodrugs thereof, wherein R7 is [ka] [ka] is selected from R8 is, -CH3, [ka] [ka] is selected from R9 is selected from H and CH3, and R 10 is selected from H and F.

[0235] In certain embodiments, the compound of Formula (III-C) has R7, R8, R9 and / or R10 as shown in Table 13. 10 It can have:

[0236] [Table 13]

[0237] In other embodiments, the compound of the present disclosure can be any one or combination of the compounds provided in Table 14.

[0238] [Table 14-1] [Table 14-2] [Table 14-3]

[0239] In some embodiments, the compound according to the present disclosure is 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidin-5-yl)oxy)phenyl)cyclobutane-1-carboxamide, [ka] In some embodiments, the compounds disclosed herein have an EC50 activity in the range of about 0.2 μM to about 1.0 μM on cancer cell viability. 50 (e.g., decreased viability, increased tumor cell killing), and can have a kinetic solubility in the range of about 0.1 μM to about 0.5 μM.

[0240] In some embodiments, the compound according to the present disclosure is 3-methoxy-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide, [ka] an analog thereof, an isomer thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a polymorph thereof, or any combination thereof.

[0241] In some embodiments, the compound is 3-methoxy-N-(3-methyl-4-((2-morpholinothiazol-5-yl)oxy)phenyl)cyclobutane-1-carboxamide, [ka] In some embodiments, the compound has an EC50 activity in the range of about 0.5 μM to 1.0 μM on cancer cell viability. 50 and has a kinetic solubility in the range of about 20.0 μM to about 25.0 μM.

[0242] In some embodiments, the compound is 3-methoxy-N-(5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide, [ka] In some embodiments, the compound has an EC50 activity in the range of about 0.5 μM to about 2.0 μM on cancer cell viability. 50 It has.

[0243] In some embodiments, the compound is 3-methoxy-N-(4-((2-(4-methoxypiperidin-1-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)cyclobutane-1-carboxamide, [ka] In some embodiments, the compound has an EC50 activity in the range of about 0.2 μM to about 1.0 μM on cancer cell viability. 50, and has a kinetic solubility in the range of about 4.0 μM to about 7.0 μM.

[0244] In some embodiments, the compound is 3-methoxy-N-(5-(4-morpholinophenoxy)thiazol-2-yl)cyclobutane-1-carboxamide, [ka] In some embodiments, the compound has an EC50 activity in the range of about 0.1 μM to about 1.0 μM on cancer cell viability. 50 and has a kinetic solubility in the range of about 20.0 μM to about 30.0 μM.

[0245] In some embodiments, the compound is N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxycyclobutane-1-carboxamide [ka] In some embodiments, the compound has an EC50 activity in the range of about 0.5 μM to about 1.0 μM on cancer cell viability. 50 It has.

[0246] In some embodiments, the compound is N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxycyclobutane-1-carboxamide [ka] In some embodiments, the compound has an EC50 activity in the range of about 0.5 μM to about 2.0 μM on cancer cell viability. 50 It has.

[0247] In some embodiments, the compound is N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide [ka] In some embodiments, the compound has an EC50 activity in the range of about 0.001 μM to about 0.02 μM for cancer cell viability. 50 , and has a kinetic solubility in the range of about 85.0 μM to about 100.0 μM.

[0248] In some embodiments, the compound is N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide [ka] In some embodiments, the compound has an EC50 activity in the range of about 45.0 μM to about 55.0 μM for cancer cell viability. 50 It has.

[0249] In some embodiments, the compound is 3-methoxy-N-(5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide, [ka] In some embodiments, the compound has an EC50 activity in the range of about 0.05 μM to about 1.0 μM on cancer cell viability. 50 , and has a kinetic solubility in the range of about 4.0 μM to about 6.0 μM.

[0250] In some embodiments, the compound is N-(5-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide [ka] In some embodiments, the compound has an EC50 activity in the range of about 0.01 μM to about 0.5 μM for cancer cell viability. 50 and has a kinetic solubility in the range of about 80.0 μM to about 90.0 μM.

[0251] In some embodiments, the compound is N-(4-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide [ka] In some embodiments, the compound has an EC50 activity in the range of about 10.0 μM to about 20.0 μM for cancer cell viability. 50 It has.

[0252] In some embodiments, the compound is 3-methoxy-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide, [ka] In some embodiments, the compound has an EC50 activity in the range of about 0.01 μM to about 1.0 μM on cancer cell viability. 50 , and has a kinetic solubility in the range of about 4.0 μM to about 25.0 μM.

[0253] In some embodiments, the compound is 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidin-5-yl)oxy)phenyl)bicyclo[1.1.1]pentane-1-carboxamide, [ka] an analog thereof, an isomer thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a polymorph thereof, or any combination thereof.

[0254] In some embodiments, the compound is 3-methoxy-N-(5-(4-morpholinophenoxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide, [ka] In some embodiments, the compound has an EC50 activity in the range of about 0.001 μM to about 0.05 μM for cancer cell viability. 50 It has.

[0255] In some embodiments, the compound is 3-methoxy-N-(5-(4-(piperidin-1-yl)phenoxy)thiazol-2-yl)cyclobutane-1-carboxamide, [ka] In some embodiments, the compound has an EC50 activity in the range of about 1.0 μM to about 2.0 μM for cancer cell viability. 50 It has.

[0256] In some embodiments, the compound is 3-methoxy-N-(5-((6-morpholinopyridin-3-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide, [ka] an analog thereof, an isomer thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a polymorph thereof, or any combination thereof.

[0257] In some embodiments, any of the formulas described herein may exclude compounds disclosed in PCT / IB2022 / 062416.

[0258] In some embodiments, the compounds of the present disclosure can include isomers of any one of the compounds disclosed herein and can be designated as being in the "cis" or "trans" configuration. According to these embodiments, the compounds of the present disclosure can be cis or trans isomers of any one of the compounds or formulas disclosed herein.

[0259] In some embodiments, compounds of the present disclosure can include asymmetrically substituted carbon atoms in the R or S configuration, where the terms "R" and "S" are as defined in the art. According to some embodiments disclosed herein, compounds having asymmetrically substituted carbon atoms with equal amounts of R and S configurations can be racemic at those carbon atoms. In certain embodiments, the present disclosure can include racemic mixtures, relative and absolute stereoisomers, and / or mixtures of relative and absolute stereoisomers.

[0260] In other embodiments, the compounds disclosed herein may be in the form of esters, such as ester prodrugs. The term "ester" herein may refer to a compound formed by modifying a functional group (e.g., a hydroxyl group, a carboxyl group, an amino group, etc.). Examples of "esters" include "esters formed with a hydroxyl group" and "esters formed with a carboxyl group." The term "ester" may refer to an ester whose ester residue is a "conventional protecting group" or a "protecting group removable in vivo by a biological method such as hydrolysis." In some embodiments, the term "conventional protecting group" may refer to a protecting group removable by a chemical method, such as hydrogenolysis, hydrolysis, electrolysis, or photolysis. In other embodiments, the term "protecting group removable in vivo by a biological method such as hydrolysis" may refer to a protecting group removable in vivo by hydrolysis, etc., to generate a free acid or a salt thereof after administration to a subject.

[0261] In certain embodiments, the compounds of the present disclosure may be in the form of pharmaceutically acceptable salts. By "salt" or "pharmaceutically acceptable salt" is meant that such salts are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, or allergic reaction, and are effective for their intended use at a reasonable benefit-risk ratio. "Pharmacologically acceptable salts" may refer to salts that may be formed when the compounds herein have an acidic group, such as carboxyl, or a basic group, such as amino or imino. In some embodiments, salts of the compounds disclosed herein can be formed with acidic groups and can include, but are not limited to, inorganic salts such as alkali metal salts, e.g., sodium, potassium, or lithium salts, alkaline earth metal salts, e.g., calcium or magnesium salts, metal salts, e.g., aluminum or iron salts, amine salts, e.g., ammonium salts, and organic salts such as t-octylamine, dibenzylamine, morpholine, glucosamine, phenylglycine alkyl ester salts, ethylenediamine, N-methylglucamine, guanidine, diethylamine, triethylamine, dicyclohexylamine, N,N'-dibenzylethylenediamine, chloroprocaine, procaine, diethanolamine, N-benzylphenethylamine, piperazine, tetramethylammonium, or tris(hydroxymethyl)aminomethane salts, as well as amino acid salts, e.g., glycine, lysine, arginine, ornithine, glutamate, aspartate.

[0262] In some embodiments, salt derivatives of the compounds disclosed herein formed with basic groups may include, but are not limited to, hydrogen halides such as hydrofluoride, hydrochloride, hydrobromide or hydroiodide, inorganic acid salts such as nitrate, perchlorate, sulfate or phosphate, lower alkane sulfonates such as methanesulfonate, trifluoromethanesulfonate or ethanesulfonate, arylsulfonates such as benzenesulfonate or p-toluenesulfonate, organic acid salts such as acetate, malate, fumarate, succinate, citrate, ascorbate, tartrate, oxalate or maleate, amino acid salts such as glycine, lysine, arginine, histidine, ornithine, glutamate or aspartate. In certain embodiments, the pharmacologically acceptable salts of the compounds disclosed herein may absorb water to form hydrates for use in the formulations disclosed herein when exposed to air or recrystallized.

[0263] In certain embodiments, compounds of the present disclosure can include, but are not limited to, compounds in solid or liquid form or state. In some embodiments, compounds of the present disclosure can be in amorphous form. In other embodiments, compounds of the present disclosure can be in crystalline form or a combination or mixture of crystalline and amorphous forms. According to some embodiments disclosed herein, solid-state compounds can exist in crystalline, powder, or amorphous form, or as a mixture thereof. In some embodiments, compounds disclosed herein in crystalline form can be used to form pharmaceutically acceptable solvates. Those skilled in the art will appreciate that pharmaceutically acceptable solvates can be formed in which solvent molecules are incorporated into the crystalline lattice during crystallization. According to some embodiments, solvates for use as disclosed herein can include non-aqueous solvents such as ethanol, isopropanol, DMSO, acetic acid, ethanolamine, and EtOAc, or can include water as the solvent incorporated into the crystalline lattice. Solvates in which water is the solvent incorporated into the crystalline lattice are typically referred to as "hydrates." Hydrates can include stoichiometric hydrates as well as compositions containing variable amounts of water. The present disclosure encompasses all such solvates known in the art.

[0264] In certain embodiments, compounds of the present disclosure can exist in various crystalline forms, including solvates thereof, and can exhibit polymorphism (e.g., the ability to appear in different crystalline structures). These different crystalline forms are referred to herein as "polymorphs." Polymorphs have the same chemical composition but differ in packing, geometric arrangement, and / or other descriptive properties of the crystalline solid state. Thus, polymorphs can have different physical properties, such as shape, density, hardness, deformability, stability, and dissolution characteristics. Polymorphs typically exhibit different melting points, IR spectra, X-ray powder diffraction patterns, and NMR signatures, which can be used for identification. In certain embodiments, compounds of the present disclosure can be polymorphs. In certain embodiments, compounds of the present disclosure can be polymorphs identified by their melting points, IR spectra, X-ray powder diffraction patterns, NMR signatures, or any combination thereof. In some embodiments, different polymorphs of the compounds of the present disclosure can be produced by changing and / or adjusting the reaction conditions and / or reagents used to make the compounds. For example (but not limited to), changes in temperature, pressure, or solvents can result in polymorphism. In some embodiments, different polymorphs of the compounds disclosed herein can spontaneously convert to another polymorph.

[0265] In certain embodiments, further to the previous paragraph, the compounds disclosed herein can have a solubility high enough to achieve the desired bioavailability and concentration in the systemic circulation for the desired pharmacological response, which can be measured using a kinetic or thermodynamic solubility approach. In some embodiments, the kinetic solubility parameters of the compounds disclosed herein can be determined. According to these embodiments, the kinetic solubility can be measured using the method described in Example 20. In some embodiments, the compounds disclosed herein can have a kinetic solubility that indicates the bioavailability of formulations containing the compounds, such as oral, inhalable, topical, subcutaneous, and / or intravenous formulations.

[0266] In other embodiments, the compounds disclosed herein can have a kinetic solubility of at least about 0.35 μM. In some embodiments, the compounds disclosed herein can have a kinetic solubility in the range of about 0.35 μM to about 92 μM (e.g., about 0.35 μM, about 1 μM, about 5 μM, about 10 μM, about 20 μM, about 30 μM, about 40 μM, about 50 μM, about 60 μM, about 70 μM, about 80 μM, and about 90 μM, or about 92 μM, or any concentration therebetween). In yet another embodiment, the compounds disclosed herein can maintain a kinetic solubility in the range of about 0.35 μM to about 92 μM (e.g., about 0.39 μM, about 1 μM, about 5 μM, about 10 μM, about 20 μM, about 30 μM, about 40 μM, about 50 μM, about 60 μM, about 70 μM, about 80 μM, and about 90 μM, or any concentration therebetween or greater). In further embodiments, the compounds disclosed herein can maintain a kinetic solubility for about 1 hour to about 48 hours or more (e.g., about 1 hour, about 3 hours, about 6 hours, about 12 hours, about 24 hours, about 36 hours, 48 ​​hours, or any time therebetween or greater than 48 hours). In some embodiments, the compounds disclosed herein can maintain a kinetic solubility in the range of about 0.35 μM to about 92 μM (e.g., about 0.39 μM, about 1 μM, about 5 μM, about 10 μM, about 20 μM, about 30 μM, about 40 μM, about 50 μM, about 60 μM, about 70 μM, about 80 μM, and about 90 μM, and about 92 μM, or any concentration therebetween or greater) for about 1 hour to about 48 hours (e.g., about 1 hour, about 3 hours, about 6 hours, about 12 hours, about 24 hours, about 36 hours, 48 ​​hours, or therebetween or longer).

[0267] In still other embodiments, the compounds disclosed herein can maintain kinetic solubility at temperatures ranging from about 4°C to about 80°C (e.g., about 4°C, about 6°C, about 8°C, about 10°C, about 20°C, about 30°C, about 40°C, about 50°C, about 60°C, about 70°C, about 80°C, or any temperature therebetween). In some embodiments, the compounds disclosed herein can maintain a kinetic solubility in the range of about 0.39 μM to about 92 μM (e.g., about 0.39 μM, about 1 μM, about 5 μM, about 10 μM, about 20 μM, about 30 μM, about 40 μM, about 50 μM, about 60 μM, about 70 μM, about 80 μM, and about 90 μM, or concentrations therebetween) at a temperature in the range of about 4°C to about 80°C (e.g., about 4°C, about 6°C, about 8°C, about 10°C, about 20°C, about 30°C, about 40°C, about 50°C, about 60°C, about 70°C, about 80°C, or temperatures therebetween).

[0268] In some embodiments, the compounds disclosed herein can maintain a kinetic solubility in the range of about 0.35 μM to about 92 μM (e.g., about 0.39 μM, about 1 μM, about 5 μM, about 10 μM, about 20 μM, about 30 μM, about 40 μM, about 50 μM, about 60 μM, about 70 μM, about 80 μM, and about 90 μM) at room temperature (i.e., 25° C.±3° C.). In some embodiments, the compounds disclosed herein can maintain a kinetic solubility in the range of about 0.35 μM to about 92 μM (e.g., about 0.39 μM, about 1 μM, about 5 μM, about 10 μM, about 20 μM, about 30 μM, about 40 μM, about 50 μM, about 60 μM, about 70 μM, about 80 μM, and about 90 μM) at room temperature (i.e., 25° C.±5° C.). A kinetic solubility in the range of about 0.39 μM to about 91.23 μM (e.g., about 0.39 μM, about 1 μM, about 5 μM, about 10 μM, about 20 μM, about 30 μM, about 40 μM, about 50 μM, about 60 μM, about 70 μM, about 80 μM, and about 90 μM) can be maintained for about 1 hour to about 48 hours (e.g., about 1 hour, about 3 hours, about 6 hours, about 12 hours, about 24 hours, about 36 hours, 48 ​​hours, or any time in between, or longer depending on the compound).

[0269] In some embodiments, the compound is configured to cross the blood-brain barrier (BBB) ​​of a human or non-human subject, hi some embodiments, the non-human subject is a livestock, companion animal, laboratory animal, or zoo animal, wild animal, reptile, fish, or bird.

[0270] In some embodiments, the compounds have an EC50 of 0.005 μM to 50 μM for inhibiting cancer cell viability or inducing cancer cell death. In some embodiments, the compounds have an EC50 of about 0.0001 μM to 50 μM for inhibiting cancer cell viability or inducing cancer cell death. In some embodiments, the compounds have an EC50 of about 0.0001 μM to 50 μM for inhibiting cancer cell viability or inducing cancer cell death. In other embodiments, the compounds have an EC50 of about 0.0001 μM to 15 μM for inhibiting cancer cell viability or inducing cancer cell death. In other embodiments, the compounds have an EC50 of about 0.0001 μM to 50 μM for inhibiting cancer cell viability or inducing cancer cell death.

[0271] In certain embodiments, compounds disclosed herein can be analyzed and have a central nervous system multiparameter optimization (CNS MPO) score. According to these embodiments, the CNS MPO score is calculated using a weighted scoring function based on six fundamental physicochemical properties: (a) lipophilicity, calculated partition coefficient (ClogP); (b) calculated partition coefficient at about pH 7.4 (ClogD); (c) molecular weight (MW); (d) topological polar surface area (TPSA); (e) number of hydrogen bond donors (HBDs); and (f) maximum basic center (pK a The CNS MPO score can be determined by an algorithm that evaluates the ability of selected compounds to cross the blood-brain barrier (BBB). In these examples, the algorithm assigns an overall score ranging from 0 to 6 to selected compounds, with a higher CNS MPO score indicating the compound's ability to cross the blood-brain barrier (BBB). In some embodiments, compounds disclosed herein can have a CNS MPO score of 4.0 or greater, indicating BBB permeability. In other embodiments, compounds disclosed herein can have a CNS MPO score of 4.0 or greater, indicating the ability to cross the BBB when used to treat a condition affecting the brain of a subject.

[0272] In some embodiments, the compound has a Papp score greater than 10. In other embodiments, the compound has an efflux ratio less than 2.0.

[0273] In some embodiments, the compound is a drug conjugate or a prodrug.

[0274] In certain embodiments, further to the preceding paragraph, the compounds disclosed herein can reduce the viability or induce cell death of at least one cancer cell in a subject by about 10.0% to about 99% or up to 100% (e.g., about 1.0% or up to 100%, about 25.0% or up to 100%, about 50.0% or up to 100%, about 75.0% or up to about 100%, about 99% to about 100%), compared to cancer cells not exposed to a compound or mixture of compounds disclosed herein or a formulation thereof. In other embodiments, the compounds disclosed herein can reduce tumor volume by about 10% to up to 100% compared to cancer cells not exposed to a compound or mixture of compounds disclosed herein or a formulation thereof. In still other embodiments, the compounds disclosed herein can reduce tumor metastasis in a subject by about 10% up to 100% compared to cancer cells not exposed to a compound or mixture of compounds disclosed herein or a formulation thereof. In some embodiments, the compounds disclosed herein can reduce cancer cell viability, reduce tumor volume, and / or reduce tumor metastasis at a median effective concentration (i.e., EC 50In some embodiments, the compounds disclosed herein may have an EC50 in the range of about 0.001 μM to about 50 μM (e.g., about 0.001 μM, about 0.1 μM, about 0.15 μM, about 0.25 μM, about 0.5 μM, about 0.75 μM, about 1.0 μM, about 2.0 μM, about 5.0 μM, about 10.0 μM, about 15.0 μM, about 20.0 μM, about 25.0 μM, about 30.0 μM, about 35.0 μM, about 40.0 μM, about 45.0 μM, up to about 50.0 μM) or about 0.0001 μM to about 15 μM for reducing or inhibiting cancer cell viability or killing cancer cells, reducing tumor volume, and / or reducing tumor metastasis. 50 It can have:

[0275] In certain embodiments, further to the preceding paragraph, the compounds disclosed herein can include cell cycle inhibitors or other antitumor or anti-neoplastic agents or compounds capable of reducing abnormal cell growth or proliferation. Cell cycle inhibitors reduce or stop cell cycle progression through various mechanisms. Cell cycle arrest can be induced at different stages, slowing the rate of cell division and reducing or inhibiting the number of actively cycling cells. In some embodiments, the compounds disclosed herein can arrest the cell cycle at G2M. In other embodiments, the compounds disclosed herein can arrest cell proliferation of at least one cancer cell or non-cancerous abnormal cell proliferation at the G2M cell cycle stage.

[0276] In other embodiments, the compounds disclosed herein are effective for treating a medical condition. In some embodiments, the compounds are effective for preventing cancer cell division. In other embodiments, the compounds are effective for inhibiting tubulin polymerization. In other embodiments, the compounds are effective for destabilizing microtubules. In other embodiments, the compounds are effective for arresting cell division in the G2 / M phase. In some embodiments, the compounds are effective for cytotoxicity against multiple cancer cell lines. In some embodiments, the compounds are effective for treating cancer or metastasis.

[0277] In some embodiments, the compounds disclosed herein are effective in treating a medical condition, hi some embodiments, the compounds are effective in treating a non-neoplastic condition.

[0278] In some embodiments, the compounds disclosed herein are effective in treating gout, familial Mediterranean fever, or onychomycosis. In other embodiments, the compounds are effective as vascular targeting agents.

[0279] In some embodiments, the compounds disclosed herein are effective in reducing tumor growth in a subject by up to 70% compared to a subject not treated with the compound.

[0280] II. Tagged Compounds / Prodrugs / Conjugated Drugs

[0281] In some embodiments, the invention features a tagged compound comprising one or more compounds. In other embodiments, the tagged compound further comprises a tag linked to the compound. In further embodiments, the tag is a fluorescent tag, a radioactive tag, biotin, or a combination thereof.

[0282] In some embodiments, the invention features prodrugs comprising one or more compounds. In other embodiments, the prodrugs further comprise an inert moiety linked to the compound. In further embodiments, the inert moiety is an ester, a carbamate, an aminoacyl ester, or a combination thereof.

[0283] In some embodiments, the present invention features a drug conjugate comprising one or more compounds. In other embodiments, the drug conjugate further comprises a targeting moiety linked to the compound. In some embodiments, the targeting moiety is an antibody, a polyethylene glycol (PEG) conjugate, a long-chain polymer, a peptide sequence, or a combination thereof.

[0284] III. Pharmaceutical Compositions

[0285] In another embodiment, the present disclosure provides a pharmaceutical composition.The pharmaceutical composition comprises at least one compound, tagged compound, prodrug, or drug conjugate according to the present disclosure for use in treating a health condition of a subject in need thereof.The compound may be, for example, as described herein.

[0286] In some embodiments, a pharmaceutical composition can comprise at least one compound disclosed herein and at least one pharmaceutically acceptable carrier. In certain embodiments, a pharmaceutical composition can comprise pharmaceutically acceptable carriers, excipients, and / or stabilizers that are non-toxic to recipients at the dosages and / or concentrations used to practice the methods disclosed herein.

[0287] In certain embodiments, the weight fraction of the excipient or combination of excipients in the composition can be about 99% or less, about 97% or less, about 95% or less, about 90% or less, about 85% or less, about 80% or less, about 75% or less, about 70% or less, about 65% or less, about 60% or less, about 55% or less, about 50% or less, about 45% or less, about 40% or less, about 35% or less, about 30% or less, about 25% or less, about 20% or less, about 15% or less, about 10% or less, about 5% or less, about 2%, or about 1% or less of the total weight of the composition.

[0288] In some embodiments, pharmaceutically acceptable excipients may include, but are not limited to, diluents, binders, fillers, buffers, pH adjusters, disintegrants, dispersants, preservatives, lubricants, taste-masking agents, flavoring agents, or coloring agents, and may be as described herein below. The concentrations and types of excipients utilized to form the pharmaceutical compositions disclosed and contemplated herein can be selected according to known principles and knowledge of pharmacy in the art.

[0289] Excipients and other adjuvants

[0290] In some embodiments, the excipient can be a diluent. The diluent can be compressible (i.e., plastically deformable) or abrasively brittle. Non-limiting examples of suitable compressible diluents include, but are not limited to, microcrystalline cellulose (MCC), cellulose derivatives, cellulose powder, cellulose esters (i.e., mixed esters of acetic and butyric acid), ethyl cellulose, methyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, sodium carboxymethyl cellulose, corn starch, phosphorylated corn starch, pregelatinized corn starch, rice starch, potato starch, tapioca starch, starch-lactose, starch-calcium carbonate, sodium starch glycolate, glucose, fructose, lactose, lactose monohydrate, sucrose, xylose, lactitol, mannitol, maltitol, sorbitol, xylitol, maltodextrin, dextran, and trehalose. Non-limiting examples of suitable abrasively friable diluents include dibasic calcium phosphate (anhydrous or dihydrate), calcium phosphate tribasic, calcium carbonate, and magnesium carbonate.

[0291] In another embodiment, the excipient can be a binder. Suitable binders include, but are not limited to, starch, pregelatinized starch, gelatin, polyvinylpyrrolidone, cellulose, methylcellulose, sodium carboxymethylcellulose, ethylcellulose, polyacrylamide, polyvinyloxazolidone, polyvinyl alcohol, C12-C18 fatty acid alcohols, polyethylene glycol, polyols, saccharides, oligosaccharides, polypeptides, oligopeptides, and combinations thereof.

[0292] In another embodiment, the excipient can be a filler.Suitable fillers include, but are not limited to, carbohydrates, inorganic compounds, and polyvinylpyrrolidone.By way of non-limiting example, the filler can be calcium sulfate, both dibasic and tribasic, starch, calcium carbonate, magnesium carbonate, microcrystalline cellulose, dibasic calcium phosphate, magnesium carbonate, magnesium oxide, calcium silicate, talc, modified starch, lactose, sucrose, mannitol, or sorbitol.

[0293] In yet another embodiment, the excipient can be a buffer. Representative examples of suitable buffers include, but are not limited to, phosphate buffer, carbonate buffer, citrate buffer, Tris buffer, polysaccharide buffer, and buffered saline (e.g., Tris-buffered saline or phosphate-buffered saline). In various embodiments, the excipient can be a pH adjuster. By way of non-limiting example, the pH adjuster can be sodium carbonate, sodium bicarbonate, sodium citrate, citric acid, or phosphoric acid. In further embodiments, the excipient can be a disintegrant. The disintegrant can be non-effervescent or effervescent. Suitable examples of non-effervescent disintegrants include, but are not limited to, starches such as corn starch, potato starch, pregelatinized and modified starches thereof, sweeteners, clays such as bentonite, microcrystalline cellulose, alginates, sodium starch glycolate, gums such as agar, guar, carob, karaya, pectin, and tragacanth. Non-limiting examples of suitable effervescent disintegration agents include sodium bicarbonate in combination with citric acid and sodium bicarbonate in combination with tartaric acid.

[0294] In yet another embodiment, the excipient may be a dispersant or dispersion enhancer. Suitable dispersants may include, but are not limited to, starch, alginic acid, polyvinylpyrrolidone, guar gum, kaolin, bentonite, purified wood cellulose, sodium starch glycolate, isomorphous silicate, and microcrystalline cellulose.

[0295] In another embodiment, the excipient can be a preservative.Non-limiting examples of suitable preservatives include antioxidants, such as BHA, BHT, vitamin A, vitamin C, vitamin E, or retinyl palmitate, citric acid, sodium citrate; chelating agents, such as EDTA or EGTA; and antibacterial agents, such as parabens, chlorobutanol or phenol.

[0296] In a further embodiment, the excipient may be a lubricant. Non-limiting examples of suitable lubricants include minerals such as talc or silica; fats such as vegetable stearin, magnesium stearate, or stearic acid.

[0297] In yet another embodiment, the excipient may be a taste-masking agent, such as a cellulose ether, polyethylene glycol, polyvinyl alcohol, a polyvinyl alcohol and polyethylene glycol copolymer, a monoglyceride or triglyceride, an acrylic polymer, a mixture of an acrylic polymer and a cellulose ether, cellulose acetate phthalate, or a combination thereof.

[0298] In an alternative embodiment, the excipient may be a flavoring agent, which may be selected from synthetic flavor oils and flavoring aromatics and / or natural oils, extracts from plants, leaves, flowers, fruits, and combinations thereof.

[0299] In yet further embodiments, the excipient may be a colorant. Suitable color additives include, but are not limited to, Food, Drug, and Cosmetic Colors (FD&C), Drug and Cosmetic Colors (D&C), or External Drug and Cosmetic Colors (Ext. D&C).

[0300] Other pharmaceutically acceptable carriers, excipients and / or stabilizers are generally non-toxic to recipients at the dosages and concentrations employed, and include, but are not limited to, buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (e.g., octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl, or benzyl alcohol; alkyl parabens, such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, histidine, arginine, lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextran; chelating agents, such as EDTA; sugars, such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions, such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants, such as TWEEN™, PLURONICS™, or polyethylene glycol (PEG).

[0301] Dosage form

[0302] In certain embodiments, the present disclosure provides dosage forms and dosage forms comprising at least one compound disclosed herein. In some embodiments, the dosage form or dosage form comprises a pharmaceutical composition or formulation comprising at least one compound according to the present disclosure. The compound and pharmaceutical composition or formulation can be as described herein and above.

[0303] In certain embodiments, the pharmaceutical compositions or formulations disclosed herein may be in unit dosage form, such as tablets, pills, capsules, powders, granules, solutions, pastes, salves or suspensions, patches (e.g., soluble or insoluble), particles, microparticles or nanoparticles, or suppositories, for oral, intravenous, subcutaneous, eye drops or other drops, or rectal administration, or administration by inhalation or insufflation. In some embodiments, to prepare solid compositions such as tablets or capsules, the primary active agent (e.g., a compound disclosed herein) can be mixed with a pharmaceutically acceptable carrier, such as a conventional tablet-forming agent, for example, corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate, or gum, and other pharmaceutically acceptable diluents, such as water, to form a solid preformulation composition containing a homogeneous mixture of a compound according to the present invention or a pharmaceutically acceptable salt thereof. When these preformulation compositions are referred to as homogeneous, it means that the active agent or compound is dispersed evenly throughout the composition, such that the composition can be easily subdivided into effective unit dosage forms, such as tablets, pills, and capsules. In some embodiments, the solid preformulation compositions disclosed herein are then formulated to contain a compound disclosed herein at a concentration of about 0.1 mg / kg to about 500 mg / kg (e.g., about 0.1 mg / kg, about 0.5 mg / kg, about 1.0 mg / kg, about 5.0 mg / kg, about 10 mg / kg, about 25 mg / kg, about 50 mg / kg, about 75 mg / kg, about 100 mg / kg, about 125 mg / kg, about 150 mg / kg, about The compound can be subdivided into unit dosage forms of the type described above containing about 175 mg / kg, about 200 mg / kg, about 225 mg / kg, about 250 mg / kg, about 275 mg / kg, about 300 mg / kg, about 325 mg / kg, about 350 mg / kg, about 375 mg / kg, about 400 mg / kg, about 425 mg / kg, about 450 mg / kg, about 475 mg / kg, about 500 mg / kg, or more, or any concentration therebetween.

[0304] In some embodiments, the tablets and / or pills disclosed herein can be coated or otherwise compounded to provide a dosage form that offers the advantage of prolonged action. In some embodiments, the tablets and / or pills herein can have an inner dosage component and an outer dosage component, the latter in the form of an envelope surrounding the former. According to embodiments herein, the two components can be separated by an enteric coating, which serves to resist disintegration in the stomach and allows the inner component to enter the duodenum intact or be released later. In certain embodiments, two or more compounds disclosed herein can be mixed together in a single dosage form. In some embodiments, the tablets and / or pills disclosed herein can contain one or more agents that can be used in such enteric layers or coatings, including, but not limited to, certain polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate. Non-limiting examples of surfactants suitable for use herein include non-ionic agents or other similar agents, such as polyoxyethylene sorbitan (e.g., Tween™ 20, 40, 60, 80, or 85) and other sorbitan (e.g., Span™ 20, 40, 60, 80, or 85). In some embodiments, compositions disclosed herein that include a surfactant can have between about 0.05% and about 5.0% of the surfactant. In some embodiments, other ingredients can be added to the pharmaceutical compositions disclosed herein, such as mannitol or other pharmaceutically acceptable vehicles as deemed appropriate.

[0305] In some embodiments, the pharmaceutical compositions disclosed herein can be tablets. According to some embodiments herein, the tablets contemplated herein can contain excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate, and glycine, disintegrants such as starch (preferably corn, potato, or tapioca starch), sodium starch glycolate, croscarmellose sodium, and certain complex silicates, and granulation binders such as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin, and gum arabic. According to some embodiments disclosed herein, the tablets contemplated herein can further contain lubricants such as magnesium stearate, stearic acid, glyceryl behenate, and talc. In some embodiments, the pharmaceutical composition can be a solid composition used as a filler in gelatin capsules. According to some embodiments disclosed herein, excipients included in the gelatin capsules contemplated herein can include lactose, starch, a cellulose, milk sugar, or high molecular weight polyethylene glycols, and the like.

[0306] In certain embodiments, the pharmaceutical compositions disclosed herein may comprise emulsions. In some embodiments, the emulsions disclosed herein can be prepared using commercially available fat emulsions, such as Intralipid™, Liposyn™, Infonutrol™, Lipofundin™, and Lipiphysan™. The active ingredients (e.g., one or more aminopeptidase inhibitors and / or one or more chemotherapeutic agents) can be dissolved in a pre-mixed emulsion composition, or can be dissolved in an emulsion formed by mixing oil (e.g., soybean oil, safflower oil, cottonseed oil, sesame oil, corn oil, or almond oil), phospholipids (e.g., egg phospholipids, soybean phospholipids, or soybean lecithin), and water. In some embodiments, other ingredients, such as glycerol or glucose, can be added to the compositions disclosed herein to adjust the tonicity of the emulsion. In other embodiments, the emulsion can contain up to 20% (w / v) oil, e.g., about 5.0% to about 20.0% (w / v). In some embodiments, the emulsion can have lipid droplets of about 0.1 μm to about 1.0 μm and / or a pH ranging from about 5.5 to about 8.0.

[0307] In certain embodiments, the pharmaceutical compositions disclosed herein can be formulated for parenteral administration or any other acceptable mode of administration, such as intravenous, intraventricular, intracisternal, or intraparenchymal injection, or a combination thereof. In some embodiments, pharmaceutical compositions herein formulated for parenteral administration can include one or more sterile liquids as pharmaceutically acceptable carriers. Non-limiting examples of sterile liquids suitable for use as pharmaceutically acceptable carriers disclosed herein can be water and oils, including, but not limited to, those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, and mineral oil. Physiological saline solutions and aqueous dextrose and polyethylene glycol (PEG) and glycerol solutions can also be used as liquid carriers, e.g., for injectable solutions. The pharmaceutical compositions disclosed herein can further include additional agents, such as preservatives, buffers, tonicity agents, antioxidants and stabilizers, non-ionic wetting or clarifying agents, thickeners, and the like. In some embodiments, the pharmaceutical compositions disclosed herein can be packaged in single-unit dosage forms or multi-dose forms.

[0308] In some embodiments, pharmaceutical compositions disclosed herein suitable for administration to a subject may include aqueous and non-aqueous sterile injection solutions. According to these embodiments, these solutions may further contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions may include, but are not limited to, suspending agents and thickening agents. Aqueous solutions may be suitably buffered (e.g., pH of about 3.0 to about 9.0, about 5.0 to about 8.5, or about 6.5 to about 8.0). The preparation of suitable parenteral formulations under sterile conditions is readily accomplished by standard pharmaceutical techniques well known to those skilled in the art.

[0309] In some embodiments, the pharmaceutical compositions described herein can further comprise one or more of an antimicrobial agent, a chemotherapeutic agent, another anti-cancer therapy, or an antibody or fragment thereof. According to these embodiments, the antimicrobial agent can be, in one example, an antiviral, bactericidal, antifungal, or antibacterial agent, or other antimicrobial agent. For example, the antimicrobial agent can be an antibacterial agent (antibiotic) such as doxycycline, tetracycline, or other antibiotic, such as a commonly applicable antibiotic.

[0310] In another embodiment, the present disclosure provides a pharmaceutical composition. The pharmaceutical composition comprises at least one compound according to the present disclosure for use in treating a condition in a subject in need thereof. The compound can be as described, for example, as described herein.

[0311] In some embodiments, a pharmaceutical composition can comprise at least one compound disclosed herein and at least one pharmaceutically acceptable carrier. In certain embodiments, a pharmaceutical composition can comprise pharmaceutically acceptable carriers, excipients, and / or stabilizers that are non-toxic to recipients at the dosages and / or concentrations used to practice the methods disclosed herein.

[0312] In certain embodiments, the weight fraction of the excipient or combination of excipients in the composition can be about 99% or less, about 97% or less, about 95% or less, about 90% or less, about 85% or less, about 80% or less, about 75% or less, about 70% or less, about 65% or less, about 60% or less, about 55% or less, about 50% or less, about 45% or less, about 40% or less, about 35% or less, about 30% or less, about 25% or less, about 20% or less, about 15% or less, about 10% or less, about 5% or less, about 2%, or about 1% or less of the total weight of the composition.

[0313] In some embodiments, pharmaceutically acceptable excipients may include, but are not limited to, diluents, binders, fillers, buffers, pH adjusters, disintegrants, dispersants, preservatives, lubricants, taste-masking agents, flavoring agents, or coloring agents, and may be as described herein below. The concentrations and types of excipients utilized to form the pharmaceutical compositions disclosed and contemplated herein can be selected according to known principles and knowledge of pharmacy in the art.

[0314] IV. Treatment method

[0315] In certain embodiments, the present disclosure provides methods for treating, preventing, reducing the occurrence of, or ameliorating a health condition in a subject having, suspected of developing, or at risk of developing a health condition. According to these embodiments, the method can include administering a therapeutically effective amount of a compound according to the present disclosure to a subject, alone or as a combination therapy, to treat, reduce the occurrence of, or ameliorate a health condition in a subject in need thereof. According to these embodiments, the compound can be in the form of a therapeutic composition comprising at least one compound according to the present disclosure, or in a dosage form comprising at least one compound according to the present disclosure. In certain embodiments, the health condition can be cancer or a non-cancerous growth. In other embodiments, the health condition can be any health condition treatable by the compounds disclosed herein. Specific embodiments of the methods and health conditions are described below. The disclosed compounds, therapeutic compositions and formulations, and dosage forms are each described above and herein.

[0316] cancer

[0317] In some embodiments, the health condition may include, but is not limited to, cancer. Thus, in certain embodiments, the methods of the present disclosure include treating, ameliorating, or reducing the risk of developing cancer in a subject. According to these embodiments, cancer may include any type of cancer or tumor. In some embodiments, cancer may be a solid tumor or a non-solid tumor, or other tumor. In other embodiments, cancer may be malignant or non-malignant, or metastatic or non-metastatic. In some embodiments, the cancer health condition may include angiogenic cancer.

[0318] Certain embodiments disclosed herein relate to treating, ameliorating, or reducing the risk of developing cancer using at least one compound disclosed herein. Non-limiting examples of cancer may include, but are not limited to, carcinoma, sarcoma, myeloma, leukemia, lymphoma, and mixed cancers. Other non-limiting examples of cancer include, but are not limited to, connective tissue, bladder cancer, breast cancer, kidney (renal) cancer, lung cancer, lymphoma, pancreatic cancer, prostate cancer, skin cancer, uterine cancer, cancer associated with other organs or tissues, and the like. According to these embodiments, the methods of the present disclosure may include the use of a compound according to the present disclosure to treat, ameliorate, prevent, or reduce the risk of developing any type of cancer.Non-limiting examples of types of cancer that may be treated, ameliorated, reduced in incidence, or prevented include, but are not limited to, adrenocortical carcinoma, AIDS-related cancer, pathogen-associated cancer (e.g., HPV), anal cancer, appendix cancer, glioblastoma, medulloblastoma, basal cell carcinoma, bladder cancer, bone cancer, brain tumor, breast cancer, bronchial adenoma / carcinoid, carcinoid tumor (childhood, gastrointestinal), carcinoma of unknown primary, cerebellar astrocytoma, cervical cancer, pediatric cancer, chronic myeloproliferative disorder, colon cancer, desmoplastic small round cell tumor, endometrial cancer, ependymoma, esophageal cancer, extrahepatic bile duct cancer, cholangiocarcinoma, eye cancer (intraocular melanoma, retinal blastoma), gallbladder cancer, gastric (stomach) cancer, gastrointestinal stromal tumor, germ cell tumors (pediatric extracranial, extragonadal, ovarian), gestational trophoblastic tumor, glioma (adult, pediatric brainstem, pediatric cerebral astrocytoma, pediatric visual pathway and hypothalamic, cerebral glioma, malignant glioma), head and neck cancer, hepatocellular (liver) cancer, hypopharyngeal cancer, laryngeal cancer, leukemia (acute lymphoblastic, acute myeloid, chronic lymphocytic, chronic myeloid, hairy cell), lip and oral cavity cancer, lung cancer (non-small cell, small cell), lymphoma (AIDS-related, Burkitt, cutaneous T-cell, Hodgkin, non-Hodgkin, primary central nervous system), macroglobulin cytopenia (Waldenstrom's disease), medulloblastoma (childhood), mesothelioma (adult malignant, childhood), metastatic squamous cell carcinoma of unknown primary, oral cancer, multiple endocrine neoplasia syndrome (childhood), multiple myeloma / plasma cell neoplasm, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative disorders, nasal cavity and paranasal sinus cancer, nasopharyngeal carcinoma, neuroblastoma, oropharyngeal cancer, osteosarcoma / malignant fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial carcinoma (surface epithelial-stromal tumor), ovarian low malignant potential tumor, pancreatic cancer, pancreatic cancer (islet cell), parathyroid carcinoma, penile cancer, pharyngeal cancer, pheochromocytoma, pineal astrocytoma, pineal germinoma, pineoblastoma, and supratentorial primitive neuroectodermal tumor (childhood), pituitary adenoma, pleuropulmonary blastoma, primary prostate cancer, rectal cancer, renal cell carcinoma (kidney cancer), rhabdomyosarcoma (childhood), salivary gland cancer, sarcoma (Ewing family of tumors, Kaposi, soft tissue, uterine), Sezary syndrome, skin cancer (non-melanoma, melanoma), skin carcinoma (Merkel cell), squamous cell carcinoma, testicular cancer, throat cancer, thymoma (childhood), thymoma and thymic carcinoma, thyroid cancer, thyroid cancer (childhood), transitional cell carcinoma of the renal pelvis and ureter, unknown primary site (adult, childhood), urethral cancer, vaginal cancer, vulvar cancer, and Wilms' tumor (childhood).

[0319] In some embodiments, non-cancerous growths can be treated with at least one compound disclosed herein to reduce progression to cancerous lesions or to suppress the growth of, for example, neoplasms or benign tumors. In accordance with these embodiments, the methods of the present disclosure can also include the use of compounds according to the present disclosure to treat, ameliorate, prevent, or reduce the risk of developing non-cancerous growths in subjects suspected of or at risk of developing any type of non-cancerous growth.

[0320] In certain embodiments, the method of the present disclosure may include the use of at least one compound disclosed herein to treat, ameliorate, prevent, or reduce the risk of developing cancer at any stage of development. For example, cancer may be stage 0 cancer, stage I, stage II, or stage III (which may be used to describe the number of tumors in a subject's body, the size of the tumor, and the extent of metastasis to nearby tissues), or stage IV cancer or metastatic cancer that has metastasized to other organs, lymph nodes, or distant parts of the body.

[0321] In some embodiments, cancers treated or prevented by the compounds and / or compositions disclosed herein may include, but are not limited to, prostate cancer, brain cancer, metastatic cancer, pancreatic cancer, lung cancer, breast cancer, kidney cancer, skin cancer, liver cancer, bladder cancer, osteosarcoma, ovarian cancer, rectal cancer, blood cancer, gastrointestinal cancer, medulloblastoma, or any combination thereof. In some embodiments, the cancer is prostate cancer. In certain embodiments, the cancer is breast cancer. In some embodiments, the cancer is bone cancer. In other embodiments, the cancer is Ewing's sarcoma. In further embodiments, the cancer is lung cancer. In yet other embodiments, the cancer is non-small cell lung cancer (NSCLC). In some embodiments, the cancer is skin cancer. In other embodiments, the cancer is melanoma.

[0322] In certain embodiments, and in all of the above paragraphs, the cancer may be brain cancer. Non-limiting examples of brain cancer may include, but are not limited to, brainstem glioma, cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumor, visual pathway and hypothalamic glioma. In some embodiments, the brain cancer may be glioblastoma, high-grade glioma, other brain cancer, or any combination thereof. In certain embodiments, the methods disclosed herein include treating, improving, or reducing the onset of glioblastoma. In other embodiments, the methods disclosed herein include treating, improving, or preventing the onset of high-grade glioma.

[0323] In some embodiments, a compound disclosed herein, such as Compound B19, Compound B120, Compound B71, Compound B122, Compound B46, Compound B121, Compound B72, Compound B130, Compound B138, Compound B118, Compound B140, Compound B137, or RGN6024, Compound B89, Compound B155, Compound B125, Compound B157, or Compound B117, or any combination thereof, or a composition or formulation thereof, can be administered to treat brain cancer. According to these embodiments, brain cancer may include, but is not limited to, brainstem glioma, cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumor, visual pathway, and hypothalamic glioma. In some embodiments, a compound disclosed herein, such as Compound B19, Compound B120, Compound B71, Compound B122, Compound B46, Compound B121, Compound B72, Compound B130, Compound B138, Compound B118, Compound B140, Compound B137, or RGN6024, Compound B89, Compound B155, Compound B125, Compound B157, or Compound B117, or any combination thereof, or a composition or formulation thereof, can be administered to treat glioblastoma, high-grade glioma, other brain cancers, or any combination thereof. In certain embodiments, a compound disclosed herein, such as Compound B19, Compound B120, Compound B71, Compound B122, Compound B46, Compound B121, Compound B72, Compound B130, Compound B138, Compound B118, Compound B140, Compound B137, or RGN6024, Compound B89, Compound B155, Compound B125, Compound B157, or Compound B117, or any combination thereof, or a composition or formulation thereof, can be administered to treat, ameliorate, or reduce the incidence of glioblastoma.In another embodiment, the methods disclosed herein include administering a compound disclosed herein, e.g., Compound B19, Compound B120, Compound B71, Compound B122, Compound B46, Compound B121, Compound B72, Compound B130, Compound B138, Compound B118, Compound B140, Compound B137, or RGN6024, Compound B89, Compound B155, Compound B125, Compound B157, or Compound B117, or any combination thereof, or a composition or formulation thereof, to treat, ameliorate, or prevent the onset of high-grade glioma.

[0324] In other embodiments, RGN6024 or a formulation thereof can be administered to treat brainstem glioma, cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumor, visual pathway and hypothalamic glioma. In some embodiments, RGN6024 or a formulation thereof can be administered to treat glioblastoma, high-grade glioma, other brain cancer, or any combination thereof. In certain embodiments, RGN6024 or a formulation thereof can be administered to treat, ameliorate, or reduce the onset of glioblastoma. In other embodiments, the methods disclosed herein include administering the compound RGN6024 or a formulation thereof to treat, ameliorate, or prevent the onset of high-grade glioma.

[0325] In some embodiments, in the preceding paragraph and the paragraph above, the cancer is a cancer that metastasizes to the brain.Non-limiting examples of cancers that are most likely to cause brain metastasis include, but are not limited to, lung cancer, breast cancer, colon cancer, kidney cancer, and melanoma.In some embodiments, the cancer is lung cancer, lung cancer that has metastasized to the brain, breast cancer, breast cancer that has metastasized to the brain, colon cancer, colon cancer that has metastasized to the brain, kidney cancer, kidney cancer that has metastasized to the brain, melanoma, or melanoma that has metastasized to the brain, or any combination thereof.In other embodiments, the cancer is non-small cell lung cancer (NSCLC) that has metastasized to the brain.In some embodiments, the cancer is breast cancer that has metastasized to the brain.

[0326] In some embodiments, a method for treating, ameliorating, or preventing cancer, metastasis, tumor formation or progression, or a combination thereof in a subject can include, but is not limited to, administering an effective amount of any of the compounds and / or pharmaceutical compositions disclosed herein. As used herein, "effective amount" refers to a dose of any compound, formulation, and / or pharmaceutical composition disclosed herein sufficient to provide a therapeutic effect to a subject having or suspected of developing cancer. In certain embodiments, a therapeutic effect on a subject having or suspected of having cancer can include alleviating the symptoms or consequences of cancer, such as reducing tumor growth, killing tumor cells, preventing or reducing the occurrence of metastasis from a primary tumor, reducing the number of tumor cells in a tumor or tumor volume, reducing or preventing tumor metastasis, inhibiting the growth or spread of tumor cells in a primary tumor, secondary tumors, and / or metastatic tumors, eliminating or killing tumor cells, etc.

[0327] Non-neoplastic conditions

[0328] In some embodiments, the health condition may include, but is not limited to, a non-neoplastic condition. A non-neoplastic condition refers to a non-cancerous, non-malignant, or benign disease or lesion that is not due to abnormal tissue growth caused by uncontrolled cell proliferation. Thus, in certain embodiments, the disclosed methods include treating, ameliorating, or reducing the risk of developing gout, familial Mediterranean fever, or onychomycosis. In other embodiments, the health condition may include, but is not limited to, a vascular disease.

[0329] subject

[0330] In some embodiments, the subject treated for any condition with any compound or composition disclosed herein may be a human. In other embodiments, the subject may be a non-human animal or other mammal. In some embodiments, the subject may be a livestock animal, a companion animal, a laboratory animal, a zoo animal, a wild animal, a reptile, a fish, or a bird. Non-limiting examples of livestock animals include, but are not limited to, pigs, cows, buffalo, goats, sheep, chickens, ducks, geese, turkeys, llamas, and alpacas. Non-limiting examples of companion animals include pets such as dogs, cats, rabbits, horses, and birds. As used herein, zoo animals may include any animal found in a zoo. Zoo and wild animals may include, but are not limited to, non-human primates, wild cats, wolves, and bears. Non-limiting examples of laboratory animals include rodents, dogs, cats, and non-human primates. In some embodiments, the subject is a human subject, eg, a fetus, infant, child, adolescent, young adult, adult, or older adult.

[0331] In some embodiments, the subject may be any subject in need of treatment or therapy. In other embodiments, the subject treated by the methods described herein may be a human subject who has, has had, is suspected of having, or is at risk of developing a health condition. In certain embodiments, the health condition is cancer. In some embodiments, the subject may have, has had, is suspected of having, or is at risk of developing cancer. In some embodiments, the subject may have, has had, is suspected of having, or is at risk of developing a solid tumor. In certain embodiments, the subject may be a mammal. In some embodiments, the subject may be human. In other embodiments, subjects in need of the methods disclosed herein may be identified by routine medical testing, e.g., clinical examination, biopsy, magnetic resonance imaging (MRI) scan, ultrasound, etc. In some embodiments, the subject treated by the methods described herein may have undergone or is undergoing anti-cancer therapy, e.g., chemotherapy, radiation therapy, immunotherapy, and / or surgery.

[0332] In some embodiments, the subject treated by the methods described herein may be a human subject who has, has had, is suspected of having, or is at risk of developing cancer. In some embodiments, the subject treated by the methods described herein may be a human subject who has, is suspected of having, or is at risk of developing, but is not limited to, prostate cancer, brain cancer, metastatic cancer, pancreatic cancer, lung cancer, breast cancer, kidney cancer, skin cancer, liver cancer, bladder cancer, osteosarcoma, prostate cancer, ovarian cancer, rectal cancer, blood cancer, skin cancer, gastrointestinal cancer, medulloblastoma, or any combination thereof. In some embodiments, the subject treated by the methods described herein may be a human subject who has, has had, is suspected of having, or is at risk of developing, adenocarcinoma of the prostate (e.g., acinar adenocarcinoma and / or prostatic ductal adenocarcinoma). In certain embodiments, the subject treated by the methods described herein may be a human subject who has, has had, is suspected of having, or is at risk of developing, astrocytoma, glioblastoma, and / or meningioma. In some embodiments, the subject treated by the methods described herein may be a human subject who has, has had, is suspected of having, or is at risk of developing exocrine pancreatic cancer (e.g., adenocarcinoma, squamous cell carcinoma, adenosquamous carcinoma, colloid carcinoma) and / or neuroendocrine pancreatic cancer. In some embodiments, the subject treated by the methods described herein may be a human subject who has, is suspected of having, or is at risk of developing ductal carcinoma in situ, invasive ductal carcinoma, breast cancer (including, but not limited to, inflammatory breast cancer and / or metastatic breast cancer). In other embodiments, the subject treated by the methods disclosed herein may be a human subject who has, is suspected of having, has had, or is at risk of developing osteosarcoma, chondrosarcoma, poorly differentiated round / spindle cell tumor, Ewing's sarcoma, hemangioendothelioma, angiosarcoma, fibrosarcoma / myofibrosarcoma, chordoma, adamantinoma, liposarcoma, leiomyosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma, synovial sarcoma, and / or malignant solitary fibrous tumor.In further embodiments, the subject treated by the methods disclosed herein may be a human subject who has, has had, is suspected of having, or is at risk of developing hepatocellular carcinoma (HCC), fibrolamellar HCC, cholangiocarcinoma, angiosarcoma, secondary liver cancer, and / or hepatoblastoma. In other embodiments, the subject treated by the methods described herein may be a human subject who has, is suspected of having, has had, or is at risk of developing urothelial carcinoma, squamous cell carcinoma, adenocarcinoma, and / or small cell carcinoma of the bladder.

[0333] In certain embodiments, the subject treated by the methods disclosed herein may be a human subject who has, has had, is suspected of having, or is at risk of developing prostate cancer. In some embodiments, the subject treated by the methods disclosed herein may be a human subject who has, has had, is suspected of having, or is at risk of developing breast cancer. In other embodiments, the subject treated by the methods disclosed herein may be a human subject who has, has had, is suspected of having, or is at risk of developing bone cancer. In still other embodiments, the subject treated by the methods disclosed herein may be a human subject who has, has had, is suspected of having, or is at risk of developing Ewing's sarcoma. In some embodiments, the subject treated by the methods disclosed herein may be a human subject who has, has had, is suspected of having, or is at risk of developing lung cancer. In further embodiments, the subject treated by the methods disclosed herein may be a human subject who has, has had, is suspected of having, or is at risk of developing non-small cell lung cancer (NSCLC). In some embodiments, the subject treated by the methods disclosed herein may be a human subject who has, has had, is suspected of having, or is at risk of developing non-small cell lung cancer (NSCLC) that has metastasized to the brain. In other embodiments, the subject treated by the methods disclosed herein may be a human subject who has, has had, is suspected of having, or is at risk of developing skin cancer. In some embodiments, the subject treated by the methods disclosed herein may be a human subject who has, has had, is suspected of having, or is at risk of developing melanoma.

[0334] Administration

[0335] The composition of the present disclosure can be administered to a subject by any method known in the art.In some embodiments, the compound disclosed herein, such as Compound B19, Compound B120, Compound B71, Compound B122, Compound B46, Compound B121, Compound B72, Compound B130, Compound B138, Compound B118, Compound B140, Compound B137, or RGN6024, Compound B89, Compound B155, Compound B125, Compound B157, or Compound B117, or any combination thereof, or their compositions or formulations can be administered parenterally, bucally, intranasally, by inhalation, intraperitoneally, intrauterinely, intratumorally, intravascularly, transdermally, subcutaneously, rectally, or intrapulmonary. Non-limiting examples of administering a compound disclosed herein, e.g., Compound B19, Compound B120, Compound B71, Compound B122, Compound B46, Compound B121, Compound B72, Compound B130, Compound B138, Compound B118, Compound B140, Compound B137, or RGN6024, Compound B89, Compound B155, Compound B125, Compound B157, or Compound B117, or any combination or composition or formulation thereof, include intravenous, intramuscular, intrathecal, or intrasternal administration, oral administration, subcutaneous administration, intratumoral, intravascular, intracerebral injection, intracisternal, intraventricular, intranasal or inhalation, parenteral, buccal, enteral, intraperitoneal, inhalable, infusion, intraocular, intravitreal, ocular, rectal, sublingual, topical, transdermal, intrapulmonary, intrauterine, vaginal, ultrasound-mediated blood-brain barrier disruption, implantable device, infusion techniques, or nanoparticle-based delivery.

[0336] In certain embodiments, the compositions disclosed herein can be administered topically.Topical administration can include the use of transdermal administration, such as transdermal patch or iontophoresis device.In some embodiments, the compositions disclosed herein can be formulated into dosage unit formulations for administration, which further include, but are not limited to, conventional non-toxic pharmaceutically acceptable adjuvants, carriers, excipients and vehicles, as described herein and above.

[0337] In certain embodiments, the dosage level of the compounds disclosed herein in the therapeutic compositions of the present disclosure can be varied to administer a compound or mixture of compounds at a concentration effective to achieve the desired therapeutic response of a particular subject. The selected dosage level can depend on various factors, including the specific compound or mixture of compounds in the composition, the activity of the therapeutic composition, the formulation, combination with other drugs or treatments, the disease and the duration of the disease, and the physical condition and previous medical history of the subject being treated. Determining the appropriate dosage for a particular situation is within the skill of a medical professional.

[0338] In some embodiments, the compositions disclosed herein are administered as needed, at or just prior to the onset of symptoms. In some embodiments, the compositions are administered periodically according to a prescribed treatment schedule. For example, the compositions of the present disclosure can be administered routinely at various intervals. In another example, the compositions can be administered daily, weekly, monthly, or over several months. In some embodiments, the compositions are administered daily. In other embodiments, the compositions are administered weekly. In yet other embodiments, the compositions are administered monthly. The compositions can also be administered every 3-6 months or longer if deemed appropriate for long-term administration or treatment. As recognized in the art, the duration of treatment can and will vary and can be determined by a medical professional.

[0339] The administration of the compositions described herein can also be carried out as part of a treatment regimen, which can include multiple instances of administering one or more compositions or a mixture of compositions.Such regimen can be designed as a method for immediately treating a condition and / or as a method (e.g., prevention) for maintaining the long-term health of a subject after being treated for a condition.For example, a treatment regimen can be designed to delay the onset of the symptoms of interest in a subject.It will be understood that determining an appropriate treatment regimen is within the skill of a medical professional.

[0340] Those skilled in the art will also understand that the compositions of the present disclosure may be co-administered with other therapeutic agents before, after, and / or during treatment with the compositions of the present disclosure. The term "co-administer" refers to the administration of two or more active ingredients simultaneously with, immediately before, or immediately after the administration of one or more additional therapies. The compounds of the present disclosure may be administered alone or co-administered to a subject with another compound or standard drug known in the art. Co-administration is meant to include simultaneous or sequential administration of the compounds individually or in combination. Dosage amounts and administration intervals can be individually adjusted to provide a level of the administered compound that is effective for the particular clinical indication being treated. This provides a treatment regimen commensurate with the severity of the individual's disease state.

[0341] In certain embodiments, a subject treated with one or more compounds or compositions disclosed herein may have completed at least one additional cancer treatment regimen, may be undergoing an additional cancer treatment regimen, or may be undergoing an additional cancer treatment regimen after surgery before or after administration of a treatment disclosed herein or a compound or composition disclosed herein. In some embodiments, the additional treatment regimen for use in treating cancer disclosed herein may include administering one or more anti-cancer therapies or treatments, such as one or more of chemotherapeutic agents, radiation therapy, small molecules, and immunomodulatory agents. In some embodiments, the anti-cancer therapy or treatment can be administered separately from the compounds disclosed herein or their derivatives. In certain embodiments, the anti-cancer therapy or treatment is administered to the subject before, during, or after administration of at least one compound disclosed herein at least once daily, every other day, every three days, twice weekly, once weekly, every two weeks, twice monthly or monthly, every other month, every six months, or other suitable dosing regimen.

[0342] In certain embodiments, the present invention further comprises the administration of one or more of an antimicrobial agent, a chemotherapeutic agent, another anti-cancer therapy, or an antibody or fragment thereof. In some embodiments, the antimicrobial agent comprises one or more of an antiviral agent, a bactericide, an antifungal agent, or an antibacterial agent, or other antimicrobial agent. In other embodiments, the antibacterial agent comprises one or more of doxycycline or tetracycline.

[0343] In some embodiments, the pharmaceutical compositions disclosed herein can be administered before, during, or after at least one or more chemotherapeutic agents. Non-limiting examples of chemotherapeutic agents can include, but are not limited to, one or more of temozolomide, lomustine, velzutiphan, cisplatin, carboplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin, doxorubicin, melphalan, roscovitine, mitomycin C, hydroxyurea, 5-fluorouracil, AraC (cytarabine), 6-mercaptopurine, 6-thioguanine, cisplatin, AraC, etoposide, gemcitabine, bortezomib, sunitinib, sorafenib, sodium valproate, HDAC inhibitors, DNA synthesis inhibitors, or dacarbazine, or combinations thereof. Non-limiting examples of HDAC inhibitors include FR01228, trichostatin A, SAHA, and / or PDX101. Non-limiting examples of DNA synthesis inhibitors include, but are not limited to, capecitabine, floxuridine, decitabine, vidaza, fludarabine, nelarabine, cladribine, clofarabine, pentostatin, thiarabine, troxacitabine, sapacitabine, or forestin. Further examples of additional chemotherapeutic agents include, but are not limited to, FLT3 inhibitors, such as semexanib (SCT5416), sunitinib (SU 11248), midostaurin (PKC412), lestatinib (CEP-701), tandutinib (MLN518), CHIR-258, sorafenib (BAY-43-9006), and / or KW-2449. Other non-limiting examples of additional chemotherapeutic agents include farnesyltransferase inhibitors, such as tipifarnib (R1 15777, Zarnestra), lonafarnib (SCH66336, Sarasar™), and / or BMS-214662. Other chemotherapeutic agents include, but are not limited to, topoisomerase II inhibitors, such as epipodophyllotoxin (etoposide), teniposide, the anthracyclines doxorubicin and / or 4-epidoxorubicin.Further non-limiting examples of additional chemotherapeutic agents include P-glycoprotein modulators, such as zosuquidar trihydrochloride (Z.3HCL), vanadate, or verapamil. Further non-limiting examples of chemotherapeutic agents include hypomethylating agents, such as 5-aza-cytidine or 2'deoxyazacytidine.

[0344] In certain embodiments, the pharmaceutical compositions disclosed herein can be administered before, during, or after at least one or more other drugs or molecules. Non-limiting examples of such other drugs or molecules include imatinib, dasatinib, nilotinib, bosutinib, regorafenib, ponatinib, sunitinib, sorafenib, erdafitinib, lenvatinib, pazopanib, afatinib, gefitinib, osimertinib, vandetanib, erlotinib, lapatinib, dacomitinib, neratinib, ribociclib, abemaciclib, palbociclib, cabozantinib, crizotinib, and axitinib. , alectinib, vemurafenib, encorafenib, dabrafenib, olaparib, rucaparib, taazoparib, niraparib, larotrectinib, entrectinib, lorlatinib, ibrutinib, cobimetinib, binimetinib, trametinib, brigatinib, gilteritinib, ceritinib, ivosidenib, carfilzomib, marizomib, alpelisib, duvelisib, copanlisib, and the like, but are not limited to these.

[0345] In other embodiments, the pharmaceutical compositions disclosed herein can be administered alone or in combination with at least one immunomodulatory agent. Non-limiting examples of such immunomodulatory agents include, but are not limited to, anti-PD1, anti-PD-L1, anti-CTLA-4, anti-OX40, anti-CD137, etc. Non-limiting examples of PD-1 inhibitors include, but are not limited to, anti-PD-1 antibodies such as pembrolizumab, nivolumab, and cemiplimab. Non-limiting examples of PD-L1 inhibitors may include atezolizumab, durvalumab, and avelumab. A non-limiting example of a CTLA-4 inhibitor is the anti-CTLA-4 antibody ipilimumab. In some embodiments, the immunomodulatory agent can be one or more inhibitors targeting checkpoint molecules selected from CD40, GITR, LAG-3, OX40, TIGIT, and TIM-3.

[0346] In certain embodiments and further in the preceding paragraph, at least one additional treatment regimen in the combination therapy disclosed herein may include administering radiation. In some embodiments, the subject may be treated with at least one radiation therapy before, during, or after administration of the compounds and / or pharmaceutical compositions disclosed herein. In other embodiments, the subject may be treated with radiation therapy at least 12 hours, about 24 hours, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 1 week, or about 2 weeks, or about 3 weeks or more before and / or after administration of the compounds and / or pharmaceutical compositions disclosed herein. In still other embodiments, the subject may be treated with radiation therapy using ionizing radiation. In some embodiments, the subject may be treated with radiation therapy delivered by a linear accelerator. In further embodiments, the subject may be treated with radiation therapy delivered directly to the tumor. In certain embodiments, radiation therapy can be delivered directly to the tumor at a dose of radiation ranging from about 2 Gy to about 150 Gy (e.g., about 2 to about 5, about 10, about 20, about 30, about 40, about 50, about 75, about 100, about 125, about 150 Gy or other suitable radiation range or dose).

[0347] In some embodiments, at least one compound disclosed herein can be administered in conjunction with radiation to treat a subject suspected of having or developing brain cancer (e.g., glioblastoma, high-grade glioma, or cancer that metastasizes to the brain). In certain embodiments, a compound disclosed herein, such as Compound B19, Compound B120, Compound B71, Compound B122, Compound B46, Compound B121, Compound B72, Compound B130, Compound B138, Compound B118, Compound B140, Compound B137, or RGN6024, Compound B89, Compound B155, Compound B125, Compound B157, or Compound B117, or any combination thereof, or a composition or formulation thereof, can be administered before, during, and / or after radiation to treat a subject suspected of having or developing brain cancer.

[0348] In certain embodiments, RGN6024 or a formulation thereof can be administered with, before, or after radiation to treat a subject suspected of having or developing brain cancer (e.g., glioblastoma, high-grade glioma, or cancer that metastasizes to the brain). In certain embodiments, RGN6024 or a formulation thereof can be administered simultaneously with or sequentially after radiation to treat a subject suspected of having or developing brain cancer or other cancer.

[0349] In some embodiments, at least one compound disclosed herein can be administered before, during, and / or after at least one chemotherapeutic agent to treat a subject suspected of having or developing brain cancer (e.g., glioblastoma, high-grade glioma, or cancer that metastasizes to the brain). In certain embodiments, a compound disclosed herein, such as Compound B19, Compound B120, Compound B71, Compound B122, Compound B46, Compound B121, Compound B72, Compound B130, Compound B138, Compound B118, Compound B140, Compound B137, or RGN6024, Compound B89, Compound B155, Compound B125, Compound B157, or Compound B117, or any combination thereof, or a composition or formulation thereof, can be administered before, during, and / or after at least one chemotherapeutic agent to treat a subject suspected of having or developing brain cancer. In some embodiments, at least one compound herein can be administered before, during, and / or after temozolomide to treat a subject having or suspected of having brain cancer (e.g., glioblastoma, high-grade glioma, or cancer that metastasizes to the brain).

[0350] In some embodiments, at least one compound herein can be administered before, during, and / or after lomustine to treat a subject with or suspected of having brain cancer. In some embodiments, at least one compound herein can be administered before, during, and / or after velzutifan to treat a subject with or suspected of having brain cancer. In other embodiments, at least one compound herein can be administered before, during, and / or after crizotinib to treat a subject with or suspected of having non-small cell lung cancer (NSCLC) that has metastasized to the brain. In yet other embodiments, at least one compound herein can be administered before, during, and / or after osimertinib to treat a subject with or suspected of having non-small cell lung cancer (NSCLC) that has metastasized to the brain.

[0351] In certain embodiments, RGN6024 or a formulation thereof can be administered with at least one chemotherapeutic agent to treat a subject having or suspected of having brain cancer (e.g., glioblastoma, high-grade glioma, or cancer that metastasizes to the brain). In some embodiments, RGN6024 or a formulation thereof can be administered before, during, and / or after temozolomide to treat a subject having or suspected of having brain cancer (e.g., glioblastoma, high-grade glioma, or cancer that metastasizes to the brain). In some embodiments, RGN6024 or a formulation thereof can be administered before, during, and / or after lomustine to treat a subject having or suspected of having brain cancer. In some embodiments, RGN6024 or a formulation thereof can be administered before, during, and / or after velzutifan to treat a subject having or suspected of having brain cancer. In other embodiments, RGN6024 or a formulation thereof can be administered before, during, and / or after crizotinib to treat a subject having or suspected of having non-small cell lung cancer (NSCLC) that has metastasized to the brain. In yet other embodiments, RGN6024 can be administered before, during, and / or after osimertinib to treat a subject having or suspected of having non-small cell lung cancer (NSCLC) that has metastasized to the brain.

[0352] In some embodiments, at least one compound disclosed herein or a formulation thereof can be administered in conjunction with radiation to treat a subject suspected of having or developing prostate cancer and / or Ewing's sarcoma. In certain embodiments, a compound disclosed herein, such as Compound B19, Compound B120, Compound B71, Compound B122, Compound B46, Compound B121, Compound B72, Compound B130, Compound B138, Compound B118, Compound B140, Compound B137, or RGN6024, Compound B89, Compound B155, Compound B125, Compound B157, or Compound B117, or any combination thereof, or a composition or formulation thereof, can be administered before, during, and / or after radiation to treat a subject suspected of having or developing prostate cancer and / or Ewing's sarcoma.

[0353] In certain embodiments, at least one compound disclosed herein may be administered with at least one inhibitor of ataxia telangiectasia and Rad3-related (ATR) protein to treat a subject who has, has had, is suspected of having, or is at risk of developing prostate cancer and / or Ewing's sarcoma. Non-limiting examples of inhibitors of ATR protein (i.e., ATR inhibitors) suitable for use herein may include Schisandrin B, NU6027, BAY 1895344, Dactolisib (NVP-BEZ235), EPT-46464, Torin 2, VE-821, AZ20, M4344 (VX-803), Ceralasertib (AZD6738), Berzosertib (M6620, VX-970), and the like. In certain embodiments, at least one compound herein can be administered before, during, and / or after Berzosertib to treat a subject having or suspected of having prostate cancer and / or Ewing's sarcoma.

[0354] In other embodiments, at least one compound disclosed herein can be administered together with at least one inhibitor of serine / threonine kinase checkpoint a 1 (CHK1) to treat a subject with or suspected of having prostate cancer and / or Ewing's sarcoma. Non-limiting examples of CHK1 inhibitors can include MK-8776 (SCH 900776), PF-477736, Prexasertib (LY2606368), Rabusertib (LY2603618), etc. In certain embodiments, at least one compound disclosed herein can be administered before, during, and / or after Rabusertib to treat a subject with or suspected of developing prostate cancer and / or Ewing's sarcoma.

[0355] In some embodiments, at least one compound disclosed herein can be administered with at least one inhibitor of PARP1 (poly(ADP)-ribose polymerase-1) to treat a subject suspected of having or developing pancreatic cancer. Non-limiting examples of PARP1 inhibitors suitable for use herein include, but are not limited to, veliparib, pamiparib (BGB-290), CEP 9722, E7016, rucaparib, niraparib, talazoparib, olaparib, and the like. In some embodiments, at least one compound disclosed herein can be administered before, during, and / or after olaparib to treat a subject suspected of having or developing pancreatic cancer.

[0356] dose

[0357] In some embodiments, the effective dose in humans is about 0.5 to 5 mg / kg. In some embodiments, the effective dose in humans is about 0.5 to 1 mg / kg. In some embodiments, the effective dose in humans is about 1 to 2 mg / kg. In some embodiments, the effective dose in humans is about 2 to 3 mg / kg. In some embodiments, the effective dose in humans is about 3 to 4 mg / kg. In some embodiments, the effective dose in humans is about 4 to 5 mg / kg. In some embodiments, the effective dose in humans is about 0.5 to 1.5 mg / kg. In some embodiments, the effective dose in humans is about 1 to 1.5 mg / kg. In some embodiments, the effective dose in humans is about 2.5 to 3 mg / kg. In some embodiments, the effective dose in humans is about 1.5 to 2.5 mg / kg. In some embodiments, the effective dose in humans is about 2 to 2.5 mg / kg. In some embodiments, the effective dose in humans is about 2.5 to 3.5 mg / kg. In some embodiments, the effective dose in humans is about 3 to 3.5 mg / kg. In some embodiments, the effective dose in humans is about 3.5 to 4.5 mg / kg. In some embodiments, the effective dose in humans is about 3.5 to 5 mg / kg. In some embodiments, the effective dose in humans is about 3.5 to 4 mg / kg. In some embodiments, the effective dose in humans is about 4 to 4.5 mg / kg. In some embodiments, the effective dose in humans is about 4.5 to 5 mg / kg. In some embodiments, the effective dose in humans is about 1 to 3 mg / kg. In some embodiments, the effective dose in humans is about 3 to 5 mg / kg. In some embodiments, the effective dose in humans is about 3 to 4.5 mg / kg.

[0358] In some embodiments, the effective dose in humans is 0.5 mg / kg. In some embodiments, the effective dose in humans is 1.0 mg / kg. In some embodiments, the effective dose in humans is 1.25 mg / kg. In some embodiments, the effective dose in humans is 1.5 mg / kg. In some embodiments, the effective dose in humans is 1.75 mg / kg. In some embodiments, the effective dose in humans is 2.0 mg / kg. In some embodiments, the effective dose in humans is 2.25 mg / kg. In some embodiments, the effective dose in humans is 2.5 mg / kg. In some embodiments, the effective dose in humans is 2.75 mg / kg. In some embodiments, the effective dose in humans is 3.0 mg / kg. In some embodiments, the effective dose in humans is 3.25 mg / kg. In some embodiments, the effective dose in humans is 3.5 mg / kg. In some embodiments, the effective dose in humans is 3.75 mg / kg. In some embodiments, the effective dose in humans is 4 mg / kg. In some embodiments, the effective dose in humans is 4.25 mg / kg. In some embodiments, the effective dose in humans is 4.5 mg / kg. In some embodiments, the effective dose in humans is 4.75 mg / kg. In some embodiments, the effective dose in humans is 5 mg / kg.

[0359] In some embodiments, the effective dose in mice is 1-25 mg / kg. In some embodiments, the effective dose in mice is 1-5 mg / kg. In some embodiments, the effective dose in mice is 5-10 mg / kg. In some embodiments, the effective dose in mice is 10-15 mg / kg. In some embodiments, the effective dose in mice is 15-20 mg / kg. In some embodiments, the effective dose in mice is 20-25 mg / kg. In some embodiments, the effective dose in mice is 1-3 mg / kg. In some embodiments, the effective dose in mice is 3-5 mg / kg. In some embodiments, the effective dose in mice is 5-8 mg / kg. In some embodiments, the effective dose in mice is 8-10 mg / kg. In some embodiments, the effective dose in mice is 10-12 mg / kg. In some embodiments, the effective dose in mice is 12-15 mg / kg. In some embodiments, the effective dose in mice is 15-17 mg / kg. In some embodiments, the effective dose in mice is 17-19 mg / kg. In some embodiments, the effective dose in mice is 17-19 mg / kg. In some embodiments, the effective dose in mice is 1-21 mg / kg. In some embodiments, the effective dose in mice is 21-23 mg / kg. In some embodiments, the effective dose in mice is 23-25 ​​mg / kg. In some embodiments, the effective dose in mice is 1-10 mg / kg. In some embodiments, the effective dose in mice is 10-20 mg / kg. In some embodiments, the effective dose in mice is 10-25 mg / kg.

[0360] In some embodiments, the effective dose in mice is 1 mg / kg. In some embodiments, the effective dose in mice is 1.5 mg / kg. In some embodiments, the effective dose in mice is 2 mg / kg. In some embodiments, the effective dose in mice is 2.5 mg / kg. In some embodiments, the effective dose in mice is 3.0 mg / kg. In some embodiments, the effective dose in mice is 3.5 mg / kg. In some embodiments, the effective dose in mice is 4.0 mg / kg. In some embodiments, the effective dose in mice is 4.5 mg / kg. In some embodiments, the effective dose in mice is 5 mg / kg. In some embodiments, the effective dose in mice is 5.5 mg / kg. In some embodiments, the effective dose in mice is 6 mg / kg. In some embodiments, the effective dose in mice is 6.5 mg / kg. In some embodiments, the effective dose in mice is 7 mg / kg. In some embodiments, the effective dose in mice is 7.5 mg / kg. In some embodiments, the effective dose in mice is 10 mg / kg. In some embodiments, the effective dose in mice is 12 mg / kg. In some embodiments, the effective dose in mice is 15 mg / kg. In some embodiments, the effective dose in mice is 20 mg / kg. In some embodiments, the effective dose in mice is 23 mg / kg. In some embodiments, the effective dose in mice is 25 mg / kg.

[0361] In some embodiments, the effective dose is administered after at least one compound is administered. In some embodiments, the effective dose is administered to the subject before at least one compound is administered. In some embodiments, the effective dose is administered to the subject while (e.g., simultaneously with) at least one compound is being administered. In some embodiments, the effective dose is administered alone.

[0362] In some embodiments, the effective dose is administered at least once daily. In some embodiments, the effective dose is administered every other day. In some embodiments, the effective dose is administered every three days. In some embodiments, the effective dose is administered twice weekly. In some embodiments, the effective dose is administered weekly. In some embodiments, the effective dose is administered every other week. In some embodiments, the effective dose is administered twice monthly. In some embodiments, the effective dose is administered monthly. In some embodiments, the effective dose is administered every other month. In some embodiments, the effective dose is administered every six months. In some embodiments, the effective dose is administered by some other suitable dosing regimen.

[0363] In certain embodiments, the present disclosure provides methods for treating, preventing, reducing the occurrence of, or ameliorating a health condition in a subject having, suspected of developing, or at risk of developing a health condition. According to these embodiments, the method can include administering a therapeutically effective amount of a compound according to the present disclosure to a subject, either alone or as a combination therapy, to treat, reduce the occurrence of, or ameliorate a health condition in a subject in need thereof. According to these embodiments, the compound can be in the form of a therapeutic composition comprising at least one compound according to the present disclosure, or in a dosage form comprising at least one compound according to the present disclosure. In certain embodiments, the health condition can be cancer or a non-cancerous growth. In other embodiments, the health condition can be any health condition treatable by the compounds disclosed herein. Specific embodiments of the methods and health conditions are described below. The disclosed compounds, therapeutic compositions and formulations, and dosage forms are each described herein and above.

[0364] How to use

[0365] In some embodiments, the present invention relates to a method of modulating abnormal cell division, the method comprising the steps of (a) identifying a cell having abnormal cell division, and (b) administering a compound or composition disclosed herein or a derivative thereof.

[0366] In some embodiments, the present invention provides a drug screening method for identifying therapeutically effective drug candidates for treating a health condition in a subject in need thereof, the method comprising: (a) identifying an in vitro or in vivo model of the health condition; (b) administering a drug candidate and RGN6024 to the in vitro or in vivo model; (c) determining the efficacy, toxicity or side effects of the drug candidate and RGN6024; and (d) comparing the efficacy, toxicity or side effects of the drug candidate and RGN6024 to identify a therapeutically effective drug candidate.

[0367] In some embodiments, the present invention provides a method for detecting target cells having aberrant cell division, the method comprising: (a) identifying a sample containing target cells having aberrant cell division; (b) performing an affinity-based assay using a small molecule conjugated to a tag to contact the target cells having aberrant cell division with the small molecule conjugated to the tag; and (c) determining whether the small molecule conjugated to the tag binds to the target cells having aberrant cell division. In some embodiments, the small molecule is RGN6024, or a compound or composition disclosed herein or a derivative thereof. In further embodiments, the affinity-based assay is an immune-based assay, a receptor-based assay, an antibody-based assay, a nanoparticle-based assay, a chemical assay, an optical assay, or a kinetic binding assay. In other embodiments, the affinity-based assay is gel electrophoresis, an enzyme-linked immunosorbent assay, an immunoblot assay, a fluorescence intensity assay, a fluorescence anisotropy assay, a fluorescence energy transfer assay, a surface plasmon resonance (SPR) assay, a light scattering assay, a forward binding assay, a dissociation assay, or a reverse binding assay. In some embodiments, the affinity-based assay is a colchicine competitive binding assay.

[0368] A method for detecting a protein expressed by a target cell having abnormal cell division, the method comprising: (a) identifying a sample containing a protein expressed by a target cell having abnormal cell division; (b) performing an affinity-based assay using a small molecule conjugated to a tag to contact the protein expressed by the target cell having abnormal cell division with the small molecule conjugated to the tag; and (c) determining whether the small molecule conjugated to the tag binds to the protein expressed by the target cell having abnormal cell division. In some embodiments, the small molecule is RGN6024, or a compound or composition disclosed herein or a derivative thereof. In further embodiments, the affinity-based assay is an immune-based assay, a receptor-based assay, an antibody-based assay, a nanoparticle-based assay, a chemical assay, an optical assay, or a kinetic binding assay. In other embodiments, the affinity-based assay is gel electrophoresis, an enzyme-linked immunosorbent assay, an immunoblot assay, a fluorescence intensity assay, a fluorescence anisotropy assay, a fluorescence energy transfer assay, a surface plasmon resonance (SPR) assay, a light scattering assay, a forward binding assay, a dissociation assay, or a reverse binding assay. In some embodiments, the affinity-based assay is a colchicine competitive binding assay.

[0369] In some embodiments, the present invention provides a method for detecting or isolating target cells having aberrant cell division, the method comprising: (a) identifying a sample containing target cells having aberrant cell division; (b) performing an affinity-based pull-down assay using a small molecule conjugated to a tag to contact the target cells having aberrant cell division with the small molecule conjugated to the tag; and (c) selectively isolating the target cells having aberrant cell division. In some embodiments, the small molecule is RGN6024, or a compound or composition disclosed herein or a derivative thereof.

[0370] In another embodiment, a method for detecting or isolating a protein expressed by a target cell having aberrant cell division comprises: (a) identifying a sample containing a protein expressed by a target cell having aberrant cell division; (b) performing an affinity-based pull-down assay using a small molecule conjugated to a tag to contact the protein expressed by the target cell having aberrant cell division with the small molecule conjugated to the tag; and (c) selectively isolating the protein expressed by the target cell having aberrant cell division. In another embodiment, the small molecule is RGN6024, or a compound or composition disclosed herein or a derivative thereof.

[0371] kit

[0372] In certain embodiments, provided herein are kits for use in the treatment, alleviation, or prevention of a target disease or condition treatable by at least one compound disclosed herein. In certain embodiments, the kits include at least one compound disclosed herein or a composition containing at least one compound, and at least one container. In some embodiments, the kits can include instructions for use according to any of the methods described herein. In other embodiments, the instructions can include instructions for administering at least one compound and / or pharmaceutical composition disclosed herein to a subject. According to embodiments herein, the kits can include instructions providing information regarding the dosage, administration schedule, and administration route for the intended treatment.

[0373] In some embodiments, the kits disclosed herein can include at least one container. According to embodiments herein, the container can be any container capable of storing at least one compound disclosed herein or at least one composition containing at least one compound, such as a tube, vial, bottle, syringe, or a unit dose, bulk package (e.g., multi-dose package), or sub-unit dose. The instructions provided in the kits of the present invention can be instructions written on a label or package insert (e.g., a paper sheet included in the kit), but machine-readable instructions (e.g., instructions recorded on a magnetic or optical storage disk) are also acceptable. The label or package insert indicates that the composition is used to treat, delay the onset of, and / or alleviate the onset of a health condition or disease contemplated herein. Instructions for performing any of the methods described herein can be provided.

[0374] The kits disclosed herein can include suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (e.g., sealed Mylar or plastic bags), etc. Packages for use in combination with specific devices, such as inhalers, nasal administration devices (e.g., atomizers), patches, or injection devices such as minipumps, are also contemplated herein. The kits can have a sterile access port (e.g., the container can be an intravenous solution bag or a vial with a stopper pierceable by a hypodermic needle). The container can also have a sterile access port (e.g., the container can be an intravenous solution bag or a vial with a stopper pierceable by a hypodermic needle). At least one active agent in the composition can be a compound disclosed herein.

[0375] Kits can optionally provide additional components, such as buffers and interpretive information. Typically, kits include a container and a label or package insert on or associated with the container. In some embodiments, the invention provides an article of manufacture comprising the contents of the above-described kit. [Example]

[0376] example

[0377] The following examples are included to illustrate certain embodiments. Those of skill in the art should understand that the techniques disclosed in the following examples represent techniques discovered to function well in the practice of the claimed methods, compositions, and devices. However, those of skill in the art should, in light of this disclosure, understand that changes can be made to some of the disclosed embodiments without departing from the spirit and scope of the embodiments of the present invention and still obtain like or similar results.

[0378] Example 1

[0379] In one exemplary method, 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidin-5-yl)oxy)phenyl)cyclobutane-1-carboxamide (compound B19) was synthesized. Figure 1 shows the chemical scheme for the synthesis of compound B19.

[0380] 3-Methoxycyclobutane-1-carboxylic acid (45.5 mg, 0.35 mmol), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (132.7 mg, 0.35 mmol), and diisopropylethylamine (200 μL, 1 mmol) were dissolved in dry tetrahydrofuran (6 mL). The resulting mixture was stirred at room temperature for 10 minutes and then cooled to 0 °C. A solution of 3-methyl-4-((2-morpholinopyrimidin-5-yl)oxy)aniline (100 mg, 0.35 mmol) in tetrahydrofuran (1 mL) was added, and the resulting mixture was stirred at room temperature for 3 hours. The mixture was then concentrated in vacuo. The residue was taken up in dichloromethane (25 mL), and the organic phase was washed with water (10 mL) and then with saturated aqueous bicarbonate (3 × 10 mL). The organic layer was collected, dried over magnesium sulfate, filtered, and concentrated under vacuum. The residue was purified by HPLC to give 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidin-5-yl)oxy)phenyl)cyclobutane-1-carboxamide (56.4 mg, 0.14 mmol, yield: 40.5%) as an off-white solid. LC-MS and HPLC analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 399.3 (M + H) + . 1 H NMR (400 MHz, DMSO-d6) δ 9.79 (s, 1H), 8.22 (s, 2H), 7.53-7.52 (m, 1H), 7.36-7.33 (m, 1H), 6.74-6.71 (m, 1H), 3.79-3.76 (m, 1H), 3.65-3.64 (m, 8H), 3.12 (s, 3H), 2.69-2.65 (m, 1H), 2.40-2.37 (m, 2H), 2.22 (s, 3H), 2.04-1.97 (m, 2H).

[0381] Example 2

[0382] In another exemplary method, 3-methoxy-N-(4-((2-(4-methoxypiperidin-1-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)cyclobutane-1-carboxamide 2-(4-methoxypiperidin-1-yl)pyrimidin-5-ol (compound B46) was synthesized. Figure 2 shows the chemical scheme for the synthesis of compound B46.

[0383] A solution of 4-methoxypiperidine (1.50 g, 13.02 mmol), 2-chloropyrimidin-5-ol (1.70 g, 13.02 mmol), and N,N-diisopropylethylamine (5.04 g, 39.06 mmol) in 1,4-dioxane (20 mL) was stirred under nitrogen at 100° C. for 16 hours. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (100 mL × 3). The organic layer was washed with brine (300 mL), dried over NaSO, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 3:1) to afford 2-(4-methoxypiperidin-1-yl)pyrimidin-5-ol (600.0 mg, 2.87 mmol) (yield = 22.0%) as a white solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 210.1 (M + H) + .

[0384] 2-(4-Methoxypiperidin-1-yl)-5-(2-methyl-4-nitrophenoxy)pyrimidine. A solution of 2-(4-methoxypiperidin-1-yl)pyrimidin-5-ol (500.0 mg, 2.39 mmol), 1-fluoro-2-methyl-4-nitrobenzene (307.7 mg, 2.39 mmol), and cesium carbonate (2.34 g, 7.17 mmol) in 1,4-dioxane (10 mL) was stirred at 100 °C for 6 hours under nitrogen. The reaction mixture was concentrated and purified by flash column chromatography (petroleum ether:ethyl acetate = 10:1) to afford 2-(4-methoxypiperidin-1-yl)-5-(2-methyl-4-nitrophenoxy)pyrimidine (460.0 mg, 1.34 mmol) (56.1% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 345.2 (M + H) + .

[0385] 4-((2-(4-Methoxypiperidin-1-yl)pyrimidin-5-yl)oxy)-3-methylaniline. A suspension of 2-(4-methoxypiperidin-1-yl)-5-(2-methyl-4-nitrophenoxy)pyrimidine (460.0 mg, 1.34 mmol) and 10% palladium-carbon (50.0 mg) in methanol (5 mL) was stirred under a hydrogen atmosphere at room temperature for 16 hours. The insoluble material was filtered off, and the filtrate was concentrated to give 4-((2-(4-methoxypiperidin-1-yl)pyrimidin-5-yl)oxy)-3-methylaniline (220.0 mg, 0.70 mmol) (52.2% yield) as a brown oil. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 315.2 (M + H) + .

[0386] 3-Methoxy-N-(4-((2-(4-methoxypiperidin-1-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)cyclobutane-1-carboxamide. A solution of 3-methoxycyclobutane-1-carboxylic acid (50.0 mg, 0.38 mmol), N,N-diisopropylethylamine (147.0 mg, 1.14 mmol), and O-benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (217.0 mg, 0.57 mmol) was stirred at 0 °C for 30 minutes. Then, 4-((2-(4-methoxypiperidin-1-yl)pyrimidin-5-yl)oxy)-3-methylaniline (120.0 mg, 0.38 mmol) was added at 0 °C, and the reaction solution was stirred at room temperature for 16 hours. The reaction mixture was washed with water, brine, dried over Na2SO4, concentrated, and purified by HPLC to give 3-methoxy-N-(4-((2-(4-methoxypiperidin-1-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)cyclobutane-1-carboxamide (54.1 mg, 34.2% yield) as a pale yellow solid. NMR and LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 427.3 (M + H) + . 1 H NMR (400 MHz, DMSO-d6):δ 9.82 (s, 1H), 8.18 (s, 2H), 7.54-7.53 (m, 1H), 7.36-7.33 (m, 1H), 6.73-6.70 (m, 1H), 4.16-4.12 (m, 2H), 3.81-3.74 (m, 1H), 3.45-3.40 (m, 1H), 3.35-3.32 (m, 2H), 3.28 (s, 3H), 3.13 (s, 3H), 2.72-2.64 (m, 1H), 2.39-2.35 (m, 2H), 2.23 (s, 3H), 2.04-1.99 (m, 2H), 1.89-1.85 (m, 2H), 1.43-1.35 (m, 2H).

[0387] Example 3

[0388] In one exemplary method, 3-methoxy-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide (compound B120) was synthesized. Figure 3 shows the chemical scheme for the synthesis of compound B120.

[0389] 5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-amine. A mixture of 5-bromothiazol-2-amine (1.14 g, 6.4 mmol), 2-chloropyrimidin-5-ol (1.1 g, 6.4 mmol), and CsCO (6.2 g, 19.1 mmol) in acetone (50 ml) was stirred at 60° C. for 4 hours. The mixture was cooled to room temperature, diluted with water (500 mL), and extracted with EtOAc (3×200 mL). The combined organic layers were washed with water and brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by column chromatography (petroleum ether:ethyl acetate=1:2) to give the desired compound (600 mg, 2.2 mmol) (35.1% yield) as a yellow solid. NMR and HPLC analyses were performed as described in the exemplary methods disclosed herein. LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 271.3 (M + H) + .

[0390] 5-((2-Morpholinopyrimidin-5-yl)oxy)thiazol-2-amine. A mixture of 5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-amine (200 mg, 0.87 mmol), morpholine (229 mg, 2.62 mmol), and CsCO (850 mg, 2.62 mmol) in acetone (10 mL) was degassed with nitrogen. The mixture was heated to 85 °C and stirred under a nitrogen atmosphere for 16 h. After completion, the reaction was cooled to room temperature, diluted with water, and extracted with EA. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by HPLC to give 5-((2-Morpholinopyrimidin-5-yl)oxy)thiazol-2-amine (100 mg, 0.36 mmol) (40.9% yield) as a yellow oil. NMR and HPLC analyses were performed as described in the exemplary methods disclosed herein. LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 280.2 (M + H) + .

[0391] 3-Methoxy-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide. 3-Methoxycyclobutane-1-carboxylic acid (46.6 mg, 0.36 mmol), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (205 mg, 0.54 mmol), and N,N-diisopropylethylamine (148 μL, 2.31 mmol) were dissolved in dry tetrahydrofuran (6 mL). The resulting mixture was stirred at room temperature for 10 minutes and then cooled to 0 °C. A solution of 5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-amine (100 mg, 0.36 mmol) in tetrahydrofuran (1 mL) was added, and the resulting mixture was stirred at room temperature for 3 hours. The mixture was then concentrated in vacuo. The resulting residue was taken up in dichloromethane (25 mL) and the organic phase was washed with water (10 mL) and then with saturated aqueous bicarbonate (3 x 10 mL). The organic layer was collected, dried over magnesium sulfate, filtered, and concentrated in vacuo. The residue was purified by HPLC to give 3-methoxy-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide (25.8 mg, 0.066 mmol, yield: 8.6%) as an off-white solid. LC-MS (ESI + ): m / z 392.2 (M + H) + NMR and HPLC analyses were performed as described in the exemplary methods herein. 1 H NMR (400 MHz, DMSO-d6) δ 8.45-8.43 (m, 2H), 7.14 (s, 1H), 3.80-3.76 (m, 1H), 3.65 (s, 8H), 3.11-3.09 (m, 3H), 2.83-2.78 (m, 1H), 2.41-2.35 (m, 2H), 2.02-1.95 (m, 2H).

[0392] Example 4

[0393] In one exemplary method, 3-methoxy-N-(5-(4-morpholinophenoxy)thiazol-2-yl)cyclobutane-1-carboxamide (compound B121) was synthesized. Figure 4 shows the chemical reaction scheme for the synthesis of compound B121.

[0394] 5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-amine. A mixture of 5-bromothiazol-2-amine (1.14 g, 6.4 mmol), 4-bromophenol (1.1 g, 6.4 mmol), and Cs2CO3 (6.2 g, 19.1 mmol) in acetone (50 ml) was stirred at 60 °C for 4 hours. The mixture was cooled to room temperature, diluted with water (500 ml), and extracted with EA (3 × 200 ml). The combined organic layers were washed with water and brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by column chromatography (petroleum ether:ethyl acetate = 1:2) to give the desired compound (600 mg, 2.2 mmol) (35.1% yield) as a yellow solid. NMR and HPLC analyses were performed as described in the exemplary methods herein. LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 271.3 (M + H) + .

[0395] N-(5-(4-Bromophenoxy)thiazol-2-yl)-3-methoxycyclobutane-1-carboxamide. 3-Methoxycyclobutane-1-carboxylic acid (286.0 mg, 2.2 mmol), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (834.4 mg, 2.2 mmol), and diisopropylethylamine (385 μL, 6 mmol) were dissolved in dry tetrahydrofuran (6 mL). The resulting mixture was stirred at room temperature for 10 minutes and then cooled to 0 °C. A solution of 5-(4-bromophenoxy)thiazol-2-amine (600 mg, 2.2 mmol) in tetrahydrofuran (1 mL) was added, and the resulting mixture was stirred at room temperature for 3 hours. The mixture was then concentrated in vacuo. The resulting residue was taken up in dichloromethane (25 mL), and the organic phase was washed with water (10 mL) and then with saturated aqueous bicarbonate (3 x 10 mL). The organic layer was collected, dried over magnesium sulfate, filtered, and concentrated in vacuo. The residue was purified by column chromatography (petroleum ether:ethyl acetate=1:1) to give N-(5-(4-bromophenoxy)thiazol-2-yl)-3-methoxycyclobutane-1-carboxamide (420 mg, 1.1 mmol, yield: 49.8%) as a yellow oil. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 383.3 (M + H) + .

[0396] 3-Methoxy-N-(5-(4-morpholinophenoxy)thiazol-2-yl)cyclobutane-1-carboxamide. A mixture of N-(5-(4-bromophenoxy)thiazol-2-yl)-3-methoxycyclobutane-1-carboxamide (420 mg, 1.1 mmol), morpholine (144 mg, 1.7 mmol), Pd2(dba)3 (201 mg, 0.22 mmol), and CyJohnPhos (116 mg, 0.33 mmol) in THF (15 mL) was degassed with nitrogen. The mixture was heated to 85 °C and stirred under a nitrogen atmosphere for 16 h. Upon completion, the reaction was cooled to room temperature, diluted with water, and extracted with EA. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by HPLC to give 3-methoxy-N-(5-(4-morpholinophenoxy)thiazol-2-yl)cyclobutane-1-carboxamide (30 mg, 7% yield) as a white solid. LC-MS (ESI + ): m / z 390.2 (M + H) + NMR and HPLC analyses were performed as described in the exemplary methods herein. 1 H NMR (400 MHz, DMSO-d6) δ 8.38 (s, 1H), 7.13 (s, 1H), 7.06-7.03 (m, 2H), 6.97-6.95 (m, 2H), 3.81-3.77 (m, 1H), 3.74-3.72 (m, 4H), 3.13-3.12 (m, 3H), 3.07-3.04 (m, 4H), 2.85-2.80 (m, 1H), 2.50-2.41 (m, 2H), 2.04-1.97 (m, 2H).

[0397] Example 5

[0398] In one exemplary method, N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxycyclobutane-1-carboxamide (compound B72) was synthesized. Figure 5 shows the chemical scheme for the synthesis of compound B72.

[0399] 2-Chloro-5-(2-methyl-4-nitrophenoxy)pyrimidine. To a solution of 2-chloropyrimidin-5-ol (839.1 mg, 6.41 mmol) in DMA (20.0 mL) was added 1-fluoro-2-methyl-4-nitrobenzene (1.01 g, 6.41 mmol) and potassium carbonate (6.22 g, 19.21 mmol), and the mixture was stirred at 100 °C overnight. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (100.0 mL × 3). The organic layer was washed with brine (150.0 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 5:1) to give 2-chloro-5-(2-methyl-4-nitrophenoxy)pyrimidine (110.5 mg, 0.31 mmol) (6.41% yield) as a yellow solid. NMR and HPLC analyses were performed as described in the exemplary methods disclosed herein. LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 266.7 (M + H) + .

[0400] 8-(5-(2-Methyl-4-nitrophenoxy)pyrimidin-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane. To a solution of 2-chloro-5-(2-methyl-4-nitrophenoxy)pyrimidine (110.5 mg, 0.31 mmol) in DMA (2.0 mL) was added 3-oxa-8-azabicyclo[3.2.1]octane (34.1 mg, 0.31 mmol) and cesium carbonate (293.4 mg, 0.91 mmol). The mixture was stirred at 150 °C for 4 h. After the reaction was quenched, the reaction mixture was extracted with ethyl acetate (40.0 mL × 3). The organic layer was washed with brine (50.0 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate=3:1) to give 8-(5-(2-methyl-4-nitrophenoxy)pyrimidin-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane (98.1 mg, 0.28 mmol) (68.29% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 343.1 (M + H) + .

[0401] 4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylaniline. A suspension of 8-(5-(2-methyl-4-nitrophenoxy)pyrimidin-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane (98.1 mg, 0.28 mmol) and 10% palladium-carbon (10.5 mg) in methanol (2.0 mL) was stirred at room temperature under a hydrogen atmosphere for 4 hours. The insoluble material was filtered off, and the filtrate was concentrated to give 4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylaniline (81.2 mg, 0.26 mmol) (92.85% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 313.1 (M + H) + .

[0402] N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxycyclobutane-1-carboxamide. To a solution of 3-methoxycyclobutanecarboxylic acid (34.1 mg, 0.26 mmol) in DMF (2.0 mL) at room temperature was added HBTU (118.2 mg, 0.31 mmol) and DIPEA (100.6 mg, 0.78 mmol). The reaction mixture was stirred for 25 min. o The mixture was stirred at RT for 10 minutes. Then, 4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylaniline (81.2 mg, 0.26 mmol) was added and stirred at room temperature overnight. After the reaction was quenched, the reaction mixture was extracted with ethyl acetate (20 mL × 3). The organic layer was washed with brine (30.0 mL), dried over NaSO, filtered, and concentrated. The residue was purified by HPLC to give N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxycyclobutanecarboxamide (25.9 mg, 0.06 mmol) (yield 23.0%) as a white solid. NMR and LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 425.3 (M + H) + . 1 H NMR (400 MHz, DMSO-d6) δ 9.90 (s, 1H), 8.23 ​​(s, 2H), 7.55 (s, 1H), 7.39-7.36 (m, 1H), 6.77 (d, J=8.8 Hz, 1H), 4.55 (s, 2H), 3.82-3.75 (m, 1H), 3.64-3.57 (m, 2H), 3.56-3.54 (m, 2H), 3.13 (s, 3H), 2.71-2.70 (m, 1H), 2.37-2.35 (m, 2H), 2.22 (s, 3H), 2.01-1.95 (m, 4H), 1.94-1.87 (m, 2H), .

[0403] Example 6

[0404] In one exemplary method, 3-methoxy-N-(3-methyl-4-((2-morpholinothiazol-5-yl)oxy)phenyl)cyclobutanecarboxamide 5 (compound B71) was synthesized. Figure 6 shows the chemical scheme for the synthesis of compound B71.

[0405] 5-(2-Methyl-4-nitrophenoxy)thiazol-2-amine. To a solution of 2-methyl-4-nitrophenol (2.05 g, 11.17 mmol) in ACN (20.0 mL) was added 5-bromothiazol-2-amine (2.01 g, 13.07 mmol) and potassium carbonate (4.91 g, 35.51 mmol), and the mixture was stirred at 80 °C for 1 h. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (100.0 mL × 3). The organic layer was washed with brine (150.0 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 3:1) to give 5-(2-methyl-4-nitrophenoxy)thiazol-2-amine (1.51 g, 6.01 mmol) (53.8% yield) as a yellow solid. NMR and HPLC analyses were performed as described in the exemplary methods disclosed herein. LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 252.3 (M + H) + .

[0406] 2-Bromo-5-(2-methyl-4-nitrophenoxy)thiazole. To a solution of 5-(2-methyl-4-nitrophenoxy)thiazol-2-amine (700.1 mg, 2.81 mmol) in ACN (10.0 mL), cupric bromide (630.2 mg, 2.81 mmol) and tert-butyl nitrite (432.5 mg, 4.21 mmol) were added, and the mixture was stirred at 25 °C overnight. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (50.0 mL × 3). The organic layer was washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 6:1) to give 2-bromo-5-(2-methyl-4-nitrophenoxy)thiazole (450.5 mg, 1.43 mmol) (50.8% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 316.2 (M + H) + .

[0407] 4-(5-(2-Methyl-4-nitrophenoxy)thiazol-2-yl)morpholine. To a solution of 2-bromo-5-(2-methyl-4-nitrophenoxy)thiazole (400.5 mg, 1.28 mmol) in DMF (10.0 mL) was added morpholine (1.12 g, 12.82 mmol) and DIPEA (489.5 mg, 3.86 mmol), and the mixture was stirred at 100 °C for 16 h. After the reaction was quenched, the reaction mixture was extracted with ethyl acetate (40.0 mL × 3). The organic layer was washed with brine (50.0 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate=3:1) to give 4-(5-(2-methyl-4-nitrophenoxy)thiazol-2-yl)morpholine (360.5 mg, 1.12 mmol) (yield 87.46%) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 322.3 (M + H) + .

[0408] 3-Methyl-4-((2-morpholinothiazol-5-yl)oxy)aniline. A suspension of 4-(5-(2-methyl-4-nitrophenoxy)thiazol-2-yl)morpholine (360.5 mg, 1.12 mmol) and 10% palladium-carbon (36.1 mg) in methanol (5.0 mL) was stirred under a hydrogen atmosphere at room temperature for 4 hours. The insoluble material was filtered off, and the filtrate was concentrated to give 3-methyl-4-((2-morpholinothiazol-5-yl)oxy)aniline (240.5 mg, 0.82 mmol) (73.6% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 292.2 (M + H) + .

[0409] 3-Methoxy-N-(3-methyl-4-((2-morpholinothiazol-5-yl)oxy)phenyl)cyclobutane-1-carboxamide. To a solution of 3-methoxycyclobutanecarboxylic acid (54.4 mg, 0.41 mmol) in DMF (2.0 mL) at room temperature was added HBTU (183.5 mg, 0.43 mmol) and DIPEA (444.3 mg, 1.23 mmol). The reaction mixture was stirred for 25 min. o The mixture was stirred at RT for 10 minutes. Then, 3-methyl-4-((2-morpholinothiazol-5-yl)oxy)aniline (120.2 mg, 0.41 mmol) was added and stirred at room temperature overnight. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (20 mL x 3). The organic layer was washed with brine (30.0 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by HPLC to give 3-methoxy-N-(3-methyl-4-((2-morpholinothiazol-5-yl)oxy)phenyl)cyclobutanecarboxamide (31.0 mg, 0.07 mmol, yield: 17.0%) as a white solid. NMR and HPLC-LC analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 404.2 (M + H) + . 1H NMR (400 MHz, DMSO-d6) δ 10.04 (s, 1H), 7.56 (s, 1H), 7.45 (d, J=9.2 Hz, 1H), 7.15 (s, 1H), 7.02 (d, J=8.8 Hz, 1H), 3.77-3.75 (m, 1H), 3.72-3.70 (m, 4H), 3.46 (s, 4H), 3.13 (s, 3H), 3.12-3.11 (m, 0.2H), 2.75-2.77 (m, 0.8H), 2.37-2.35 (m, 2H), 2.24(s, 3H), 2.01-1.98 (m, 2H).

[0410] Example 7

[0411] In one exemplary method, 3-methoxy-N-(5-((6-morpholinopyridin-3-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide (compound B126) was synthesized. Figure 7 shows the chemical scheme for the synthesis of compound B126.

[0412] 5-(benzyloxy)-2-chloropyridine. A solution of 6-chloropyridin-3-ol (1 g, 7.7 mmol) and CsCO (7.5 g, 23.1 mmol) in ACN (30 mL) was added, followed by (bromomethyl)benzene (1.59 g, 9.3 mmol) and the mixture was stirred at 80°C for 4 hours. The reaction mixture was extracted with ethyl acetate (300 mL x 3). The organic layer was washed with brine (300 mL), dried over NaSO, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 10:1) to obtain 5-(benzyloxy)-2-chloropyridine (1.66 g, 7.7 mmol) (yield = 99%) as a brown solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 220.67 (M + H) + .

[0413] 4-(5-(benzyloxy)pyridin-2-yl)morpholine. A solution of 5-(benzyloxy)-2-chloropyridine (1.66 g, 7.7 mmol), morpholine (1.4 g, 16.2 mmol), Pd(dba) (743 mg, 0.82 mmol), t-BuONa (2.3 g, 24.3 mmol), and BINAP (1.0 g, 1.62 mmol) in toluene (30 mL) was stirred at 100° C. for 16 hours under N. After quenching the reaction, the mixture was purified by flash column chromatography (petroleum ether:ethyl acetate=8:1) to afford 4-(5-(benzyloxy)pyridin-2-yl)morpholine (2.0 g, 7.5 mmol) (97% yield) as a yellow oil. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 271.32 (M + H) + .

[0414] 6-Morpholinopyridin-3-ol. To a solution of 4-(5-(benzyloxy)pyridin-2-yl)morpholine (2.0 g, 7.4 mmol) in MeOH (20 mL) at room temperature, Pd / C (200 mg) was added and stirred under H2 at room temperature for 4 hours. After quenching the reaction, the reaction mixture was filtered and washed with DCM / MeOH (300 mL). The organic layer was concentrated to give 6-morpholinopyridin-3-ol (1.3 g, 7.2 mmol) (97% yield). LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 181.2 (M + H) + .

[0415] tert-Butyl (5-((6-morpholinopyridin-3-yl)oxy)thiazol-2-yl)carbamate. A solution of 6-morpholinopyridin-3-ol (629 mg, 3.49 mmol), tert-butyl (5-bromothiazol-2-yl)carbamate (650 mg, 2.3 mmol), and Cs2CO3 (2.2 g, 6.9 mmol) in DMF (20 mL) was stirred at 100 °C for 1 h. The reaction mixture was extracted with ethyl acetate (300 mL × 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate=3:1) to give tert-butyl (5-((6-morpholinopyridin-3-yl)oxy)thiazol-2-yl)carbamate (80 mg, 0.21 mmol) (9% yield). LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 379.45 (M + H) + .

[0416] 5-((6-morpholinopyridin-3-yl)oxy)thiazol-2-amine. To a solution of tert-butyl-(5-((6-morpholinopyridin-3-yl)oxy)thiazol-2-yl)carbamate (80 mg, 0.21 mmol) in DCM (10 mL) at room temperature was added TFA (2 mL) and stirred at room temperature for 4 hours. The mixture was concentrated to give 5-((6-morpholinopyridin-3-yl)oxy)thiazol-2-amine (58 mg, 0.2 mmol) (99% yield) as a yellow oil. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 279.48 (M + H) + .

[0417] 3-Methoxy-N-(5-((6-morpholinopyridin-3-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide. A solution of 5-((6-morpholinopyridin-3-yl)oxy)thiazol-2-amine (58 mg, 0.2 mmol), 3-methoxycyclobutane-1-carboxylic acid (55 mg, 0.43 mmol), DIPEA (81.2 mg, 0.63 mmol), and HBTU (95 mg, 0.25 mmol) in DMF (2 mL) was stirred at room temperature for 2 hours. The reaction mixture was extracted with ethyl acetate (20 mL × 3). The organic layer was washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by HPLC to give 3-methoxy-N-(5-((6-morpholinopyridin-3-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide (36.5 mg, 0.09 mmol) (46% yield). NMR and HPLC-LC analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 391.2 (M + H) + . 1 H NMR (400 MHz, DMSO-d6) δ 12.13 (s, 1H), 7.98-7.97 (m, 1H), 7.83-7.79 (m, 1H), 7.22-7.18 (m, 2H), 3.81-3.79 (m, 1H), 3.74-3.71 (m, 4H), 3.57-3.55 (m, 4H), 3.12-3.10 (m, 3H), 2.85-2.84 (m,1H), 2.40-2.37 (m, 2H), 2.02-1.98(m, 2H).

[0418] Example 8

[0419] In one exemplary method, 3-methoxy-N-(5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide (compound B122) was synthesized. Figure 8 shows the chemical scheme for the synthesis of compound B122.

[0420] tert-Butyl (5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)carbamate. To a solution of tert-butyl (5-bromothiazol-2-yl)carbamate (1.02 g, 3.61 mmol) in ACN (10.0 mL), 2-chloropyrimidin-5-ol (470.5 mg, 3.61 mmol) and cesium carbonate (2.91 g, 8.93 mmol) were added, and the mixture was stirred at 60 °C for 16 h. After the reaction was quenched, the reaction mixture was extracted with ethyl acetate (100 mL × 3). The organic layer was washed with brine (150.0 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate=3:1) to give tert-butyl (5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (420.2 mg, 1.28 mmol) (35.48% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 329.7 (M + H) + .

[0421] 5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-amine. To a solution of tert-butyl (5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (420.1 mg, 1.27 mmol) in MeOH (4.0 mL) was added HCl / dioxane (2.0 mL), and the solution was stirred at 25° C. for 4 hours. The solution was concentrated to give 5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-amine (365.2 mg, 1.59 mmol) (100% yield) as a yellow solid. NMR and HPLC analyses were performed as described in the exemplary methods disclosed herein. LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 229.7 (M + H) + .

[0422] N-(5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxycyclobutane-1-carboxamide. To a solution of 3-methoxycyclobutanecarboxylic acid (205.3 mg, 1.57 mmol) in DMF (2.0 mL) at room temperature was added HBTU (714.5 mg, 1.88 mmol) and DIPEA (607.2 mg, 4.71 mmol). The reaction mixture was stirred for 25 min. o The mixture was stirred at RT for 10 minutes. 5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-amine (365.2 mg, 1.59 mmol) was then added and stirred at room temperature overnight. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (20.0 mL x 3). The organic layer was washed with brine (30.0 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by HPLC to give N-(5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxycyclobutanecarboxamide (220.3 mg, 0.64 mmol) (41.2% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 341.7 (M + H) + .

[0423] 3-Methoxy-N-(5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)cyclobutane-1-carboxamide. To a solution of N-(5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxycyclobutanecarboxamide (100.2 mg, 0.31 mmol) in dioxane (4.0 mL), piperidine (75.2 mg, 0.93 mmol) and DIPEA (116.5 mg, 0.93 mmol) were added, and the solution was stirred at 100° C. for 2 h. After the reaction was quenched, the reaction mixture was extracted with ethyl acetate (50.0 mL × 3). The organic layer was washed with brine (100.0 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by HPLC to give 3-methoxy-N-(5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)cyclobutanecarboxamide (45.7 mg, 0.12 mmol) (37.85% yield) as a yellow solid. NMR and LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 390.2 (M + H) + . 1 H NMR (400 MHz, DMSO-d6) δ 12.02 (s, 1H), 8.37 (s, 2H), 7.13 (s, 1H), 3.97-3.95 (m, 0.2H), 3.80-3.78 (m, 0.8H), 3.77-3.69 (m, 4H), 3.21-3.20 (m, 0.2H), 3.19 (s, 3H), 2.82-2.80 (m, 0.8H), 2.372.36 (m, 2H), 2.13-2.12 (m, 0.4H), 2.02-1.94 (m, 1.6H), 1.62-1.61(m, 2H), 1.54-1.51(m, 4H).

[0424] Example 9

[0425] In one exemplary method, 3-methoxy-N-(5-(4-(piperidin-1-yl)phenoxy)thiazol-2-yl)cyclobutane-1-carboxamide (compound B125) was synthesized. Figure 9 shows the chemical reaction scheme for the synthesis of compound B125.

[0426] tert-Butyl (5-(4-bromophenoxy)thiazol-2-yl)carbamate. To a solution of 4-bromophenol (1.01 g, 5.83 mmol) in DMA (10.0 mL) was added tert-butyl (5-bromothiazol-2-yl)carbamate (1.63 g, 5.83 mmol) and potassium carbonate (2.41 g, 17.49 mmol), and the mixture was stirred at 100 °C overnight. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (50.0 mL × 3). The organic layer was washed with brine (100.0 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate=6:1) to give tert-butyl (5-(4-bromophenoxy)thiazol-2-yl)carbamate (260.5 mg, 0.70 mmol) (yield 12.0%) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 372.2 (M + H) + .

[0427] tert-Butyl (5-(4-(piperidin-1-yl)phenoxy)thiazol-2-yl)carbamate. To a solution of tert-butyl (5-(4-bromophenoxy)thiazol-2-yl)carbamate (260.5 mg, 0.70 mmol) in dioxane (5.0 mL) was added piperidine (119.5 mg, 1.41 mmol), tri-tert-butylphosphine (28.5 mg, 0.14 mmol), Pd2(dba)3 (81.2 mg, 0.07 mmol), and sodium tert-butoxide (202.3 mg, 2.12 mmol), and the mixture was stirred at 100 °C for 16 h. After the reaction was quenched, the reaction mixture was extracted with ethyl acetate (40.0 mL × 3). The organic layer was washed with brine (100.0 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate=2:1) ​​to give tert-butyl (5-(4-(piperidin-1-yl)phenoxy)thiazol-2-yl)carbamate (121.3 mg, 0.32 mmol) (46.15% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 376.5 (M + H) + .

[0428] 5-(4-(piperidin-1-yl)phenoxy)thiazol-2-amine. To a solution of tert-butyl (5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (121.5 mg, 0.32 mmol) in DCM (3.0 mL) was added TFA (1.0 ml), and the solution was stirred at 25° C. for 4 hours. The solution was concentrated to give 5-(4-(piperidin-1-yl)phenoxy)thiazol-2-amine (165.2 mg, 0.59 mmol) (100% yield) as a brown oil. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 276.5 (M + H) + .

[0429] 3-Methoxy-N-(5-(4-(piperidin-1-yl)phenoxy)thiazol-2-yl)cyclobutane-1-carboxamide. To a solution of 3-methoxycyclobutanecarboxylic acid (76.5 mg, 0.59 mmol) in DMF (2.0 mL) at room temperature was added HBTU (272.8 mg, 0.72 mmol) and DIPEA (234.2 mg, 1.82 mmol). The reaction mixture was stirred for 25 min. o The mixture was stirred at RT for 10 minutes. 5-(4-(piperidin-1-yl)phenoxy)thiazol-2-amine (165.2 mg, 0.59 mmol) was then added and stirred at room temperature overnight. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (20.0 mL x 3). The organic layer was washed with brine (30.0 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by HPLC to give 3-methoxy-N-(5-(4-(piperidin-1-yl)phenoxy)thiazol-2-yl)cyclobutanecarboxamide (28.1 mg, 0.33 mmol) (56.0% yield) as a white solid. NMR and LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 425.3 (M + H) + . 1 H NMR (400 MHz, DMSO-d6) δ 12.08 (s, 0.5H), 7.90 (d, J=9.2 Hz, 2H), 7.31-7.29 (m, 3H), 3.81-3.80 (m, 1H), 3.51-3.49 (m, 4H), 3.12 (s, 3H), 2.85-2.83 (m, 1H), 2.44-2.39 (m, 2H), 2.03-1.97 (m, 6H), 1.66 (s, 2H).

[0430] Example 10

[0431] In one exemplary method, N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxycyclobutane-1-carboxamide (compound B130) was synthesized. Figure 10 shows the chemical scheme for the synthesis of compound B130.

[0432] tert-Butyl (5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)carbamate. A solution of tert-butyl (5-bromothiazol-2-yl)carbamate (1 g, 3.5 mmol) and Cs2CO3 (3.4 g, 10.5 mmol) in ACN (20 mL) was added to the mixture, followed by 2-chloropyrimidin-5-ol (600 mg, 4.6 mmol) and stirring at 70 °C for 4 h. The reaction mixture was extracted with ethyl acetate (100 mL × 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate=5:1) to give tert-butyl (5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (600 mg, 1.8 mmol) (50% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 329.72 (M + H) + .

[0433] tert-Butyl (5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)carbamate. A solution of tert-butyl (5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (200 mg, 0.6 mmol), 3-oxa-8-azabicyclo[3.2.1]octane (68 mg, 0.6 mmol), and DIPEA (235 mg, 1.8 mmol) in 1,4-dioxane (20 mL) was stirred at 100 °C for 16 h. The reaction mixture was extracted with ethyl acetate (50 mL × 3). The organic layer was washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate=3:1) to give tert-butyl (5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (140 mg, 0.34 mmol) (57% yield). LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 406.51 (M + H) + .

[0434] 5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-amine. To a room temperature solution of tert-butyl (5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (140 mg, 0.34 mmol) in DCM (10 mL) was added TFA (2 mL) and stirred at room temperature for 4 hours. The mixture was concentrated to give 5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-amine (107 mg, 0.32 mmol) (99% yield). LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI +): m / z 406.51 (M + H) + .

[0435] N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxycyclobutane-1-carboxamide. A solution of 5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-amine (107 mg, 0.32 mmol), 3-methoxycyclobutane-1-carboxylic acid (63 mg, 0.49 mmol), DIPEA (123 mg, 0.96 mmol), and HBTU (145 mg, 0.38 mmol) in DMF (2 mL) was stirred at room temperature for 2 hours. The reaction mixture was extracted with ethyl acetate (20 mL × 3). The organic layer was washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by preparative HPLC to give N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxycyclobutane-1-carboxamide (19.3 mg, 0.04 mmol) (14% yield). NMR and HPLC analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 418.2 (M + H) + . 1 H NMR (400 MHz, DMSO-d6) δ 12.1 (s, 1H), 8.42 (s, 2H), 7.16 (s, 1H), 4.53 (s, 2H), 3.99-3.76 (m, 1H), 3.62-3.54 (m, 4H), 3.12-3.11 (m, 3H), 2.84-2.80 (m, 1H), 2.40-2.37 (m,2H), 1.97-1.93 (m, 4H),1.88-1.86(m, 2H).

[0436] Example 11

[0437] In one exemplary method, 3-methoxy-N-(5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide (compound B140) was synthesized. Figure 11 shows the chemical reaction scheme for the synthesis of compound B140.

[0438] tert-Butyl (5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)carbamate. To a solution of tert-butyl (5-bromothiazol-2-yl)carbamate (400 mg, 1.44 mmol) in DMF (10 mL) was added 2-chloropyrimidin-5-ol (222.9 mg, 1.73 mmol) and Cs2CO3 (1.4 g, 4.32 mmol). The mixture was stirred for 10 minutes. o C for 12 hours. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (100 mL x 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by flash column chromatography (petroleum ether:ethyl acetate = 3:1) to give tert-butyl (5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (250.0 mg, 0.76 mmol) (yield 53.0%) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 328.04 (M + H) + .

[0439] tert-Butyl (5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)carbamate. To a solution of tert-butyl (5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (250 mg, 0.76 mmol) in DMF (5 mL) was added piperidine (331.2 mg, 3.8 mmol) and DIPEA (294.1 mg, 2.28 mmol). The mixture was stirred for 10 minutes. oC for 12 hours. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (100 mL x 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by flash column chromatography (petroleum ether:ethyl acetate = 3:2) to give tert-butyl (5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (180.0 mg, 0.48 mmol) (yield 62.9%) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 377.15 (M + H) + .

[0440] 5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-amine. A solution of tert-butyl (5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (180 mg, 0.47 mmol), TFA (3 mL) in DCM (5 mL) was stirred at 0° C. for 3 hours. After quenching the reaction, the solution was concentrated to give 5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-amine (120.0 mg, 0.43 mmol) (99.01% yield) along with the TFA salt as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 277.1 (M + H) + .

[0441] 3-Methoxy-N-(5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide. To a solution of 5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-amine (120 mg, 0.43 mmol) in DMF (3 mL) was added 3-methoxybicyclo[1.1.1]pentane-1-carboxylic acid (50 mg, 0.35 mmol), HBTU (159.9 mg, 0.42 mmol), and DIPEA (136.2 mg, 1.06 mmol). The mixture was stirred at room temperature for 3 hours. After the reaction was quenched, the reaction mixture was extracted with ethyl acetate (100 mL × 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by HPLC to give 3-methoxy-N-(5-((2-(piperidin-1-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide (63.9 mg, 0.16 mmol) (45.3% yield) as a yellow solid. NMR and LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 401.2 (M + H) + . 1 H NMR (400 MHz, DMSO-d6) δ 12.12 (s, 1H), 8.38 (s, 2H), 7.16 (s, 1H), 3.72-3.69 (m, 4H), 3.21 (s, 3H), 2.17 (s, 6H), 1.63-1.59 (m, 2H), 1.54-1.48 (m, 4H).

[0442] Example 12

[0443] In one exemplary method, 3-methoxy-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide (RGN6024) was synthesized. Figure 12A shows the chemical scheme for the synthesis of RGN6024.

[0444] 2-Morpholinopyrimidin-5-ol. A solution of 2-chloropyrimidin-5-ol (150 g, 1.15 mol) in morpholine (400 mL) was stirred at 80° under a nitrogen atmosphere for 16 hours. The reaction mixture was then concentrated under vacuum. The residue was purified by column chromatography (petroleum ether:ethyl acetate=1:2) to give 2-morpholinopyrimidin-5-ol (150 g). LC-MS (ESI + ): m / z 182.1 [M + H] + .

[0445] 5-((2-Morpholinopyrimidin-5-yl)oxy)thiazol-2-amine. To a solution of 2-morpholinopyrimidin-5-ol (80.0 g, 442 mmol) and 5-bromothiazol-2-amine hydrobromide (229 g, 883 mmol) in N,N-dimethylformamide (500 mL) was added CsCO (431 g, 1.33 mol). The resulting solution was stirred overnight at 55 °C under a nitrogen atmosphere. After cooling to room temperature, the mixture was poured into water (2 L) and then extracted with ethyl acetate (300 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness in vacuo. The residue was purified by column chromatography (petroleum ether:ethyl acetate = 1:1) to give 5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-amine (50.0 g) as a yellow solid. LC-MS (ESI + ): m / z 280.1 [M + H] + .

[0446] 3-Methoxy-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide. To a solution of 5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-amine (50.0 g, 0.18 mol), 3-methoxybicyclo[1.1.1]pentane-1-carboxylic acid (25.0 g, 0.18 mol), and N,N-diisopropylethylamine (69.0 g, 0.54 mol) in DMF (300 mL) was added HATU (136.1 g, 0.36 mol). The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 16 hours. The mixture was then poured into water (1.5 L) and then extracted with ethyl acetate (250 mL × 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness in vacuo. The residue was purified by column chromatography (petroleum ether:ethyl acetate=1:2) to give 3-methoxy-N-(5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide (24.1 g) as an off-white solid. LC-MS (ESI + ): m / z 404.0 [M + H] + 1H NMR (400 MHz, DMSO-d6) δ 12.14 (s, 1H), 8.44 (s, 2H), 7.18 (m, 1H), 3.66 (s, 8H), 3.20 (s, 3H), 2.17 (s, 6H).

[0447] Example 13

[0448] In one exemplary method, N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide (compound B138) was synthesized. Figure 13 shows the chemical reaction scheme for the synthesis of compound B138.

[0449] tert-Butyl (5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)carbamate. To a solution of tert-butyl (5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (101.7 mg, 0.31 mmol) in DMF (5 mL) was added piperidine (36.9 mg, 0.31 mmol) and DIPEA (119.9 mg, 0.93 mmol). The mixture was stirred for 100 min. o C for 12 hours. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (100 mL x 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by flash column chromatography (petroleum ether:ethyl acetate = 3:2) to give tert-butyl (5-((2-morpholinopyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (90.0 mg, 0.22 mmol) (yield 75.4%) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 405.15 (M + H) + .

[0450] 5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-amine. A solution of tert-butyl (5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (90 mg, 0.22 mmol) and TFA (3 mL) in DCM (5 mL) was stirred at 0 °C for 3 h. After the reaction was quenched, the solution was concentrated to give 5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-amine (65.1 mg, 0.21 mmol) (97.0% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 305.1 (M + H) +.

[0451] N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide. To a solution of 5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-amine (65.1 mg, 0.21 mmol) in DMF (3 mL) was added 3-methoxybicyclo[1.1.1]pentane-1-carboxylic acid (40 mg, 0.28 mmol), HBTU (128.3 mg, 0.34 mmol), and DIPEA (109.1 mg, 0.85 mmol). The mixture was stirred at room temperature for 3 hours. After the reaction was quenched, the reaction mixture was extracted with ethyl acetate (100 mL x 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by HPLC to give N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide (33.8 mg, 0.08 mmol) (yield 28.0%) as a yellow solid. NMR and LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 428.2 (M + H) + . 1 H NMR (400 MHz, DMSO-d6) δ 12.14 (s, 1H), 8.43 (s, 2H), 7.20 (s, 1H), 4.54 (s, 2H), 3.62-3.55 (m, 4H), 3.21 (s, 3H), 2.18 (s, 6H), 1.97-1.94 (m, 2H), 1.89-1.87 (m, 2H).

[0452] Example 14

[0453] In one exemplary method, N-(5-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide (compound B137) was synthesized. Figure 14 shows the chemical reaction scheme for the synthesis of compound B137.

[0454] tert-Butyl (5-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)thiazole-2-yl)carbamate. A solution of tert-butyl (5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (222.3 mg, 0.68 mmol), 2-oxa-7-azaspiro[4.4]nonane (129 mg, 1.0 mmol), and K2CO3 (279.8 mg, 2.03 mmol) in dioxane (5 mL) was heated to 100 °C under nitrogen for 3 h. After the reaction was quenched, the reaction mixture was extracted with ethyl acetate (100 mL × 3). The organic layer was washed with brine (300 mL) and dried over Na2SO4 to give tert-butyl (5-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (180.0 mg, 0.43 mmol) (63.3% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 419.2 (M + H) + .

[0455] 5-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)thiazol-2-amine. A solution of tert-butyl (5-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)carbamate (180.0 mg, 0.43 mmol) and TFA (3 mL) in DCM (5 mL) was stirred at 0° C. for 3 hours. After the reaction was quenched, the solution was concentrated to give 5-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)thiazol-2-amine (132.2 mg, 0.41 mmol) (96.4% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 319.1 (M + H) + .

[0456] N-(5-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide. To a solution of 5-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)thiazol-2-amine (132.2 mg, 0.41 mmol) in DMF (3 mL) was added 3-methoxybicyclo[1.1.1]pentane-1-carboxylic acid (58.2 mg, 0.41 mmol), HBTU (233.1 mg, 0.62 mmol), and DIPEA (158.7 mg, 1.23 mmol). The mixture was stirred at room temperature for 3 hours. After the reaction was quenched, the reaction mixture was extracted with ethyl acetate (100 mL x 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by HPLC to give N-(5-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)thiazol-2-yl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide (33.0 mg, 0.08 mmol) (18.6% yield) as a yellow solid. NMR and LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 433.2 (M + H) + . 1 H NMR (400 MHz, DMSO-d6) δ 12.10 (s, 1H), 8.39 (s, 2H), 7.14 (s, 1H), 3.82-3.78 (m, 2H), 3.60-3.58 (m, 2H), 3.56-3.52 (m, 2H), 3.49-3.46 (m, 2H), 3.20 (s, 3H), 2.17 (s, 6H), 1.98-1.94 (m, 2H), 1.93-1.86 (m, 2H).

[0457] Example 15

[0458] In one exemplary method, 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidin-5-yl)oxy)phenyl)bicyclo[1.1.1]pentane-1-carboxamide (compound B89) was synthesized. Figure 15 shows the chemical scheme for the synthesis of compound B89.

[0459] 2-Morpholinopyrimidin-5-ol. A solution of 2-chloropyrimidin-5-ol (800.0 mg, 6.13 mmol), morpholine (5.3 g, 61.29 mmol), and DIPEA (2.4 g, 18.39 mmol) in 1,4-dioxane (10 mL) was stirred at 100°C for 3 hours. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (100 mL x 3). The organic layer was washed with brine (300 mL), dried over NaSO, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 3:1) to give 2-morpholinopyrimidin-5-ol (650.0 mg, 3.59 mmol) (58.6% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 182.1 (M + H) + .

[0460] 4-(5-(2-Methyl-4-nitrophenoxy)pyrimidin-2-yl)morpholine. To a solution of 2-morpholinopyrimidin-5-ol (300.0 mg, 1.66 mmol) in DMF (4 mL) was added 1-fluoro-2-methyl-4-nitrobenzene (257.5 mg, 1.66 mmol) and Cs2CO3 (1.6 g, 4.98 mmol) at room temperature. The reaction mixture was stirred at room temperature for 3 hours. After the reaction was quenched, the reaction mixture was extracted with ethyl acetate (100 mL × 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate=3:1) to give 4-(5-(2-methyl-4-nitrophenoxy)pyrimidin-2-yl)morpholine (200.0 mg, 0.63 mmol) (51.46% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 317.3 (M + H) + .

[0461] 3-Methyl-4-((2-morpholinopyrimidin-5-yl)oxy)aniline. To a room temperature solution of 4-(5-(2-methyl-4-nitrophenoxy)pyrimidin-2-yl)morpholine (200.0 mg, 0.63 mmol) in MeOH (4 mL) was added Pd / C (20 mg). The reaction mixture was stirred at room temperature under a hydrogen atmosphere for 3 hours. After filtration and concentration, the crude product was purified by flash column chromatography (petroleum ether:ethyl acetate=2:1) ​​to give 3-methyl-4-((2-morpholinopyrimidin-5-yl)oxy)aniline (120.0 mg, 0.42 mmol) (66.7% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 287.1 (M + H) + .

[0462] 3-Methoxy-N-(3-methyl-4-((2-morpholinopyrimidin-5-yl)oxy)phenyl)bicyclo[1.1.1]pentane-1-carboxamide. To a room temperature solution of 3-methyl-4-((2-morpholinopyrimidin-5-yl)oxy)aniline (120.0 mg, 0.42 mmol) in DMF (3 mL) was added 3-methoxybicyclo[1.1.1]pentane-1-carboxylic acid (59.64 mg, 0.42 mmol), HBTU (238.77 mg, 0.63 mmol), and DIPEA (162.54 mg, 1.26 mmol). The reaction mixture was stirred at room temperature for 4 hours. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (100 mL × 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by HPLC to give 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidin-5-yl)oxy)phenyl)bicyclo[1.1.1]pentane-1-carboxamide (78.5 mg, 0.19 mmol) (45.53% yield) as a yellow oil. NMR and LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 411.4 (M + H) + . 1 H NMR (400 MHz, DMSO-d6) δ 9.53 (s, 1H), 8.23 ​​(s, 2H), 7.55 (d, J = 2.4 Hz, 1H), 7.41-7.38 (m, 1H), 6.74-6.72 (m, 1H), 3.68-3.63 (m, 8H), 3.22 (s, 3H), 2.23 (s, 3H), 2.13 (s, 6H).

[0463] Example 16

[0464] In one exemplary method, 3-methoxy-N-(5-(4-morpholinophenoxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide (compound B155) was synthesized. Figure 16 shows the chemical reaction scheme for the synthesis of compound B155.

[0465] 4-Morpholinophenol. A solution of 4-bromophenol (1.0 g, 5.78 mmol), morpholine (2.5 g, 28.90 mmol), Cy-JohnPhos (405.0 mg, 1.156 mmol), Pd2(dba)3 (264.5 mg, 0.29 mmol), and NaHMDS (8.8 mL, 17.34 mmol) in DMF (10 mL) was stirred at 100 °C for 6 h under N2 protection. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (100 mL × 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate=3:1) to give 4-morpholinophenol (535.1 mg, 2.99 mmol) (51.66% yield) as a pink solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 180.1 (M + H) + .

[0466] tert-Butyl (5-(4-morpholinophenoxy)thiazol-2-yl)carbamate. To a room temperature solution of 4-morpholinophenol (260.0 mg, 1.45 mmol) in DMF (5 mL) was added tert-butyl (5-bromothiazol-2-yl)carbamate (443.3 mg, 1.59 mmol) and Cs2CO3 (1.4 g, 4.35 mmol). The mixture was heated at 80 °C for 4 h. After the reaction was quenched, the reaction mixture was extracted with ethyl acetate (100 mL × 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate=3:1) to give tert-butyl (5-(4-morpholinophenoxy)thiazol-2-yl)carbamate (80.0 mg, 0.22 mmol) (yield 14.63%) as a yellow oil. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 378.14 (M + H) + .

[0467] 5-(4-morpholinophenoxy)thiazol-2-amine. To a solution of tert-butyl (5-(4-morpholinophenoxy)thiazol-2-yl)carbamate (80.0 mg, 0.22 mmol) in DCM (3 mL) at 0° C., TFA (1 mL) was added. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated to give 5-(4-morpholinophenoxy)thiazol-2-amine (50.0 mg, 0.18 mmol) (81.9% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 278.1 (M + H) + .

[0468] 3-Methoxy-N-(5-(4-morpholinophenoxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide. To a room temperature solution of 5-(4-morpholinophenoxy)thiazol-2-amine (50.0 mg, 0.18 mmol) in DMF (3 mL) was added 3-methoxybicyclo[1.1.1]pentane-1-carboxylic acid (25.6 mg, 0.18 mmol), HATU (102.6 mg, 0.27 mmol), and DIPEA (69.7 mg, 0.54 mmol). The reaction mixture was stirred at room temperature for 4 hours. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (100 mL × 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by HPLC to give 3-methoxy-N-(5-(4-morpholinophenoxy)thiazol-2-yl)bicyclo[1.1.1]pentane-1-carboxamide (7.8 mg, 0.02 mmol) (10.8% yield) as a yellow oil. NMR and LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 401.8 (M + H) + . 1H NMR (400 MHz, DMSO-d6) δ 10.81 (s, 1H), 7.15 (s, 1H), 7.03 (d, J = 9.2 Hz, 2H), 6.95 (d, J = 9.2 Hz, 2H), 3.74-3.71 (m, 4H), 3.20 (s, 3H), 3.06-3.04 (m, 4H), 2.17 (s, 6H).

[0469] Example 17

[0470] In one exemplary method, N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide (compound B118) was synthesized. Figure 17 shows the chemical scheme for the synthesis of compound B118.

[0471] 2-Chloro-5-(2-methyl-4-nitrophenoxy)pyrimidine. A solution of 1-fluoro-2-methyl-4-nitrobenzene (5.00 g, 33.26 mmol), 2-chloropyrimidin-5-ol (5.05 g, 38.71 mmol), and potassium carbonate (13.36 g, 96.78 mmol) in DMA (70 mL) was stirred at 100°C under nitrogen for 8 hours. The reaction mixture was extracted with ethyl acetate (100 mL x 3). The organic layer was washed with brine (100 mL), dried over NaSO, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 1:1) to give 2-chloro-5-(2-methyl-4-nitrophenoxy)pyrimidine (220.0 mg, 0.83 mmol) (2.5% yield) as a yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 266.0 (M + H) +

[0472] 8-(5-(2-Methyl-4-nitrophenoxy)pyrimidin-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane. A mixture of 2-chloro-5-(2-methyl-4-nitrophenoxy)pyrimidine (200.0 mg, 0.75 mmol), 3-oxa-8-azabicyclo[3.2.1]octane hydrochloride (338.0 mg, 2.26 mmol), and cesium carbonate (1.22 g, 3.75 mmol) in DMA (2 mL) was treated under microwave conditions (150° C., 3 h) and then cooled to room temperature. The reaction mixture was purified by flash column chromatography to give 8-(5-(2-methyl-4-nitrophenoxy)pyrimidin-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane (210.0 mg, 0.61 mmol) (81.3% yield) as a yellow solid. NMR and HPLC analyses were performed as described in the exemplary methods disclosed herein. LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 343.1 (M + H) +

[0473] 4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylaniline. A suspension of 8-(5-(2-methyl-4-nitrophenoxy)pyrimidin-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane (210.0 mg, 0.61 mmol) and 10% palladium-carbon (21.0 mg) in methanol (5 mL) was stirred at room temperature under a hydrogen atmosphere for 16 hours. The insoluble material was filtered off, and the filtrate was concentrated to give 4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylaniline (150.0 mg, 0.48 mmol) (78% yield) as a pale yellow solid. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 313.2 (M + H) + .

[0474] N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide. A solution of 3-methoxybicyclo[1.1.1]pentane-1-carboxylic acid (50.0 mg, 0.38 mmol), N,N-diisopropylethylamine (68.0 mg, 0.48 mmol), and O-benzotriazole-N,N,N',N'-tetramethyl-uronium-hexafluorophosphate (235.0 mg, 0.62 mmol) was stirred at room temperature for 30 minutes. Then, 4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylaniline (150.0 mg, 0.48 mmol) was added at room temperature, and the reaction solution was stirred at room temperature for 16 hours. The reaction mixture was washed with water and brine, dried over Na2SO4, concentrated, and purified by HPLC to give N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide (152.9 mg, 0.35 mmol) (73% yield) as a yellow solid. NMR and LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 437.6 (M + H) + . 1 H NMR (400 MHz, DMSO-d6) δ 9.57 (s, 1H), 8.24 (s, 2H), 7.56 (d, J = 2.4 Hz, 1H), 7.44 -7.38 (m, 1H), 6.77 (d, J = 8.4 Hz, 1H), 4.57-4.52 (m, 2H), 3.64-3.62 (m, 2H), 3.57-3.54 (m, 2H), 3.22 (s, 3H), 2.23 (s, 3H), 2.13 (s, 6H), 1.98-1.93 (m, 2H), 1.92-1.82 (m, 2H).

[0475] Example 18

[0476] In one exemplary method, N-(4-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide (compound B117) was synthesized. Figure 18 shows the chemical scheme for the synthesis of compound B117.

[0477] 2-(2-Oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-ol. A solution of 2-chloropyrimidin-5-ol (300.0 mg, 2.31 mmol), 2-oxa-7-azaspiro[4.4]nonane (881.0 mg, 6.93 mmol), and DIPEA (894.0 mg, 6.93 mmol) in 1,4-dioxane (8 mL) was stirred at 80°C under nitrogen for 16 hours. After the reaction was quenched, the reaction mixture was extracted with ethyl acetate (50 mL × 3). The organic layer was washed with brine (100 mL), dried over NaSO, filtered, and concentrated. The crude product was purified by flash column chromatography (DCM:MeOH=93:7) to give 2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-ol (150 mg, 0.68 mmol) (29.3% yield) as a brown oil. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 222.1 (M + H) + .

[0478] 7-(5-(2-Methyl-4-nitrophenoxy)pyrimidin-2-yl)-2-oxa-7-azaspiro[4.4]nonane. A solution of 2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-ol (150 mg, 0.68 mmol), 1-fluoro-2-methyl-4-nitrobenzene (105.0 mg, 0.68 mmol), and CsCO (665.0 mg, 3.04 mmol) in DMF (3 mL) was stirred at room temperature for 3 hours. After the reaction was quenched, the reaction mixture was extracted with ethyl acetate (50 mL × 3). The organic layer was washed with brine (100 mL), dried over NaSO, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate=1:2) to give 7-(5-(2-methyl-4-nitrophenoxy)pyrimidin-2-yl)-2-oxa-7-azaspiro[4.4]nonane (150 mg, 0.42 mmol) (61.9% yield) as a yellow oil. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 357.3 (M + H) +

[0479] 4-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)-3-methylaniline. A suspension of 7-(5-(2-methyl-4-nitrophenoxy)pyrimidin-2-yl)-2-oxa-7-azaspiro[4.4]nonane (150 mg, 0.42 mmol) in methanol (10 mL) was stirred under a hydrogen atmosphere at room temperature for 16 hours. The insoluble material was filtered off, and the filtrate was concentrated to give 4-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)-3-methylaniline (120.0 mg, 0.37 mmol) (81.8% yield) as a black oil. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 327.6 (M + H) + .

[0480] N-(4-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide. A solution of 3-methoxybicyclo[1.1.1]pentane-1-carboxylic acid (53.0 mg, 0.37 mmol), N,N-diisopropylethylamine (143.0 mg, 1.11 mmol), and O-benzotriazole-N,N,N',N'-tetramethyl-uronium-hexafluorophosphate (182.0 mg, 0.48 mmol) was stirred at room temperature for 30 minutes. Then, 4-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)-3-methylaniline (120.0 mg, 0.37 mmol) was added at room temperature, and the reaction solution was stirred at room temperature for 16 hours. The reaction mixture was washed with water and brine, dried over Na2SO4, concentrated, and purified by HPLC to give N-(4-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidin-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide (145.5 mg, 0.32 mmol) (87.3% yield) as a yellow solid. NMR and LC-MS analyses were performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 451.5 (M + H) + . 1 H NMR (400 MHz, DMSO-d6) δ 9.55 (s, 1H), 8.23 ​​(s, 2H), 7.55 (d, J = 2 Hz, 1H), 7.39 (d, J = 8.8 Hz, 1H), 6.69 (d, J = 8.8 Hz, 1H), 3.83-3.76 (m, 2H), 3.62-3.58 (m, 2H), 3.57-3.49 (m, 2H), 3.48-3.46 (m, 2H), 3.21 (s, 3H), 2.24 (s, 3H), 2.12 (s, 6H), 2.00-1.96 (m, 2H), 1.93-1.88 (m, 2H).

[0481] Example 19

[0482] In another exemplary method, median effective concentrations (EC ) of several compounds disclosed herein and their isomers were measured in various cancer cell lines (e.g., human cancer cell lines). 50 ) was determined. Briefly, cells from one prostate cancer cell line (22RV1) and four glioblastoma cell lines (U87, LN-18, LN-229, and T98G) were plated in multiwell plates. After 24 hours, increasing concentrations of one of the compounds disclosed herein were added to the wells. After 72 hours of treatment, cell viability was assessed using alamarBlue® (Invitrogen), and fluorescence was read using a CLARIOstar Plus Multilabel Plate Reader (BMG Labtech). Samples were normalized to untreated controls. EC 50 Values ​​were determined using GraphPad Prism. EC values ​​for compounds tested in each cell line were 50 are provided in Tables 15 and 16. In the tables, "Inactive" means less than 50% inhibition at the highest concentration tested, 50 μM, and "NT" means not tested.

[0483] [Table 15-1] [Table 15-2] [Table 15-3] [Table 15-4]

[0484] [Table 16]

[0485] Example 20

[0486] In another exemplary method, the central nervous system multiparameter optimization (CNS MPO) score, kinetic solubility, and cell permeability of selected compounds were determined. The CNS MPO scores of the compounds disclosed herein and their isomers were calculated using an algorithm employing a weighted scoring function that evaluates six key physicochemical properties for blood-brain barrier (BBB) ​​penetration (clogP, clogD, MW, TPSA, HBD, and pKa). CNS MPO scores range from 0 to 6.0, and a score of ≥ 4.0 was used as a cutoff to select compounds with a high likelihood of accumulating in the CNS. The kinetic solubility of a compound is the maximum solubility of the compound's fastest precipitating species. Kinetic solubility was determined by preparing concentrated stock solutions of the compounds disclosed herein and their isomers in an organic solvent (DMSO), subsequently mixing the solution with aqueous PBS buffer, and then filtering. The filtrate was tested, and kinetic solubility was quantified using an HPLC-MS calibration curve. Cell permeability was measured using the apparent permeability coefficient (P app ) and efflux ratio (P app BA / P app AB The apparent permeability coefficient (Papp) was determined by calculating the following equation:

number

[0487] CNS MPO score, kinetic solubility, and P of the tested compounds app and efflux ratios are shown in Table 17 ("ND" means not determined).

[0488] [Table 17]

[0489] Example 21

[0490] In another exemplary method, the effect of the compounds of the present disclosure on tubulin polymerization was determined. Note that tubulin polymerization is an important system or target for controlling abnormal cell growth or proliferation (e.g., in tumor growth or proliferation). Figures 19A and 19B show the effect of 5 μM concentrations of compounds B130, B144, and B147 (Figure 19A) and compounds B137, B138, and RGN6024 (Figure 19B) on tubulin polymerization. Colchicine and nocodazole are positive controls, and DMSO is the negative control used in these tests. The results demonstrate that many compounds, including compounds B137, B138, and RGN6024, significantly inhibit tubulin polymerization.

[0491] These results were further confirmed using a colchicine competitive binding assay (Figure 21) and an N,N'-ethylene-bis(iodoacetamide) (EBI) competitive assay (Figure 22). Microtubule-targeting agents that bind to the colchicine site of tubulin are of interest in antitumor therapy because they act as antimitotic agents. The results demonstrate that RGN6024 significantly inhibits microtubule assembly (Figure 21). Nocodazole was used as a positive control. Compound B138 was demonstrated to be an inhibitor of EBI binding using the MCF7 cell line, demonstrating binding of the compound at the colchicine binding site (Figure 22). Colchicine (COL) was used as a positive control, and vinblastine was used as a negative control.

[0492] Example 22

[0493] In another exemplary method, the levels of compound B138 and RGN6024 in plasma after oral administration were determined. For these studies, ICR1 male mice were used as an acceptable mouse model. Compound B138 or RGN6024 was then orally administered to mice as a solution in 30% hydroxypropyl-β-cyclodextrin in saline. A single dose of 30 mg / kg of compound B138 or RGN6024 was administered in this manner. The concentrations of compound B138 or RGN6024 in plasma were measured using HPLC-MS quantification (Figure 23 and Figure 24). The levels of compound B138 and RGN6024 in plasma were higher than the concentrations required to kill cancer cells in vitro.

[0494] Example 23

[0495] In another exemplary method, the level of RGN6024 in brain tissue after oral administration was measured. For these studies, the ICR1 male mouse strain was used as an acceptable mouse model. RGN6024 was then orally administered to mice as a solution in 30% hydroxypropyl-β-cyclodextrin in saline. In this manner, the concentration of RGN6024 in brain tissue administered with a single dose of 30 mg / kg of RGN6024 was measured using HPLC-MS quantification (Figure 25). The levels of RGN6024 in plasma and brain were higher than the concentration required to kill cancer cells in vitro.

[0496] Example 24

[0497] In another exemplary method, RGN6024 was tested in an LN-18 xenograft model in CB17 SCID female mice. For this study, CB17 SCID mice were implanted with LN-18 cancer cells. Tumor volumes were measured on days 2, 4, 6, 8, 10, 13, and 15. RGN6024 was orally administered at a dose of 15 mg / kg (from days 1 to 5) and at a dose of 7.5 mg / kg (from days 9, 10, and 12 to 15). The concentrations of RGN6024 in plasma, brain, and tumor tissue at 30 and 90 minutes were measured using HPLC-MS quantification (Figure 26A). The amount of compound RGN6024 in plasma, brain, and tumor was greater than the level required to kill LN-18 cancer cells.

[0498] Example 25

[0499] In another exemplary method, RGN6024 was further characterized as shown and described herein below.

[0500] As disclosed herein, it has been discovered that compounds of the present application can be used to treat conditions such as cancer. Furthermore, it has been observed that many of the compounds target cell cycle elements and induce cancer cell death. For example, RGN6024 has been observed to target and bind to tubulin. Briefly, in one experiment, U-87 cells were plated at a density of 4000 cells / well in a black 96-well plate (PhenoPlate, Perkin Elmer) and grown overnight. These cells were treated with compounds for 24 hours and then fixed with 4% paraformaldehyde for 20 minutes. After washing with PBS, the cells were permeabilized with FoxP3 perm buffer (BD Biosciences) for 10 minutes at room temperature. The cells were incubated overnight with anti-TUBB3 (1:1000; Tuj1, STEMCELL Technologies) antibody, followed by incubation with a secondary antibody for 1 hour. Nuclear DNA was labeled with Hoechst. Representative images (40x) were obtained using an automated high-content imaging microscope (Operetta, Perkin Elmer).

[0501] Separate experiments have shown that RGN6024 has an improved ability to cross the blood-brain barrier compared to other known products on the market, many of which fail to enter the brain and others have a much lower success rate at crossing the BBB than that observed herein for RGN6024 and related compounds.

[0502] In one study, RGN6024 was tested against placebo and temozolomide, a standard-of-care therapy for the treatment of glioblastoma. A gold-standard cell line representing the drug-resistant glioblastoma, LN-18, was grown in mice, and the mice were divided into three cohorts. For these studies, CB17 SCID mice were implanted with LN-18 cancer cells. Tumor volume was measured every two days. RGN6024 was administered orally at a dose of 15 mg / kg (days 1-5) and at a dose of 7.5 mg / kg (days 9, 10, and 12-15). Temozolomide was administered orally at a dose of 25 mg / kg daily on a 7-day schedule (5 days on and 2 days off). Mice treated with RGN6024 experienced an average reduction in tumor growth rate of approximately 54% compared to placebo, demonstrating improved outcomes compared to standard-of-care treatment for glioblastoma. Furthermore, in a brain cancer model, 50% of mice in the RGN6024 group experienced significant tumor shrinkage, a remarkable result compared to standard therapeutic molecules such as temozolomide, which only showed an approximately 40% reduction in tumor volume.

[0503] Current FDA-approved therapies related to tubulin interactions have been observed to bind to tubulin in large clefts on the protein's surface. Therefore, these drugs are large in size. Regarding brain penetration, the larger the molecule, the less likely it is to penetrate the brain. Therefore, large molecules have difficulty crossing the blood-brain barrier. When these large molecules cross the blood-brain barrier or are administered directly to the brain, they are often actively removed from the brain by molecular pumps designed to remove toxins. Therefore, these molecules not only have brain-barrier penetration issues, but are often pumped back out. This increases the dose required to overcome these issues, reducing efficacy and efficiency and increasing the likelihood of toxicity issues.

[0504] As disclosed herein, using a different approach, some molecules in this application have been engineered to bind to small clefts on the surface of the tubulin protein. This change allows for smaller molecule sizes, properties that affect and improve brain penetration and circumvent efflux pumps that would otherwise remove these molecules from the brain. For example, RGN6024, like many of the related molecules disclosed herein, is half the size of FDA-approved therapeutic agents.

[0505] In another exemplary method, to further study RGN6024 and its treatment of brain cancer, LN-18 xenograft tumors and brains were excised from mouse brain cancer models after the final administration. This was done at two time points: half of the mice at 30 minutes and the other half at 90 minutes. The concentration of the target drug in the brain reflected the concentration of the target drug in the tumor at both time points. As demonstrated herein, the brain concentration of RGN6024 exceeds the amount required to shrink tumors in animals, thus allowing for more effective treatment of these tumors using lower doses.

[0506] In another exemplary method, RGN6024 was tested in various high-grade glioma lines, as well as breast, lung, and melanoma tumors with a propensity for brain metastasis. A median efficacy of 91 nM was observed. In comparison, the brain penetration level of RGN6024 is 100-fold higher than the concentration required to see an effective response in these cancer models.

[0507] Example 26

[0508] In another exemplary method, patients with treatment-resistant glioblastoma can be or will be treated with once-daily oral administration of RGN6024 as a single agent or in combination with other treatments. Brain and tumor biopsy or other minimally invasive analysis, such as blood analysis or other body fluid analysis, confirms that RGN6024 achieves high concentrations directly in tumors and brains. Notably, patients will also experience significant tumor shrinkage. Surprisingly, patients continue to survive significantly longer than 5 years (when currently available treatments are used, the patient survival rate for this period is only 5%).

[0509] Example 27

[0510] In another exemplary method, the use of one or more of the compounds disclosed herein as leads for drug screening using artificial intelligence (AI) has been further characterized as shown and as described herein below.

[0511] In some embodiments, the invention features a system for generating one or more candidate compounds using artificial intelligence (AI). The one or more candidate compounds may be derived from RGN6024, may incorporate RGN6024, or a combination thereof. In some embodiments, the system may include a processor configured to execute computer-readable instructions and a memory component communicatively coupled to the processor. The memory component may include an AI model including one or more neural networks trained with a training dataset including chemical training data. Training the AI ​​model may include providing the training dataset as input to the AI ​​model. The AI ​​model may be trained to generate one or more candidate compounds as output. The memory component may further include computer-readable instructions. The computer-readable instructions may include inputting chemical data into the AI ​​model. The chemical data may include data specific to RGN6024. The computer-readable instructions may further include generating one or more candidate compounds by the AI ​​model.

[0512] In some embodiments, the invention features a method for generating one or more candidate compounds using artificial intelligence (AI). The one or more candidate compounds may be derived from RGN6024, may incorporate RGN6024, or a combination thereof. The method may include providing an AI model including one or more neural networks trained with a training dataset including chemical training data. Training the AI ​​model may include providing the training dataset as input to the AI ​​model. The AI ​​model may be trained to generate one or more candidate compounds as output. The method may further include inputting chemical data including data specific to RGN6024 into the AI ​​model and generating one or more candidate compounds by the AI ​​model.

[0513] In some embodiments, the one or more candidate compounds may include one or more candidate drug compounds. In some embodiments, the data specific to RGN6024 may include one or more structural properties, one or more physical properties, one or more interactions between RGN6024 and one or more other chemical compounds, one or more molecular pathways, one or more molecular cellular profiles, or a combination thereof.

[0514] In some embodiments, the invention features a method for training an artificial intelligence (AI) model to generate one or more candidate compounds. The one or more candidate compounds may be derived from RGN6024, may incorporate RGN6024, or a combination thereof. The AI ​​model may include one or more neural networks. The training method may include providing as input to the AI ​​model a training dataset including chemical training data specific to RGN6024. The method may further include inputting chemical data to the AI ​​model, the chemical data including data specific to RGN6024. The AI ​​model may be trained to generate one or more candidate compounds as output.

[0515] In some embodiments, the chemical training data can include data regarding one or more chemical compounds. The data regarding the one or more chemical compounds can include one or more structural properties of the one or more chemical compounds, one or more physical properties of the one or more chemical compounds, one or more interactions between one or more chemical compounds, one or more molecular pathways of the one or more chemical compounds, one or more molecular cellular profiles of the one or more chemical compounds, or a combination thereof.

[0516] In some embodiments, the chemistry training data may include one or more vector representations of one or more chemical compounds. In some embodiments, the chemistry training data may include one or more knowledge graphs representing one or more chemical compounds. In some embodiments, the chemistry training data may include any one-dimensional or multi-dimensional representation of one or more chemical compounds.

[0517] In some embodiments, the chemical training data may include one or more training labels associated with one or more chemical compounds. In some embodiments, the one or more training labels may include one or more label elements having predetermined values. In some embodiments, the one or more training labels may include bioassay results, toxicity, cross-reactivity, pharmacokinetics, pharmacodynamics, bioavailability, solubility data, or a combination thereof.

[0518] In some embodiments, the chemical data can include data about one or more chemical compounds. The data about the one or more chemical compounds can include one or more structural properties of the one or more chemical compounds, one or more physical properties of the one or more chemical compounds, one or more interactions between one or more chemical compounds, one or more molecular pathways of the one or more chemical compounds, one or more molecular cellular profiles of the one or more chemical compounds, or a combination thereof.

[0519] In some embodiments, the chemical data may include one or more vector representations of one or more chemical compounds. In some embodiments, the chemical data may include one or more knowledge graphs representing one or more chemical compounds. In some embodiments, the chemical data may include any one-dimensional or multi-dimensional representation of one or more chemical compounds.

[0520] In some embodiments, the chemical data may include one or more labels associated with one or more chemical compounds. In some embodiments, the one or more labels may include one or more label elements having predetermined values. In some embodiments, the one or more labels may include bioassay results, toxicity, cross-reactivity, pharmacokinetics, pharmacodynamics, bioavailability, solubility data, or a combination thereof.

[0521] The AI ​​model may be stored, trained, and / or executed entirely on the user's computer, partially on the user's computer, as a standalone software package, partially on the user's computer and partially on a remote computer, or entirely on a remote computer or server. The AI ​​model may be stored in the form of program code, as described above. The one or more neural networks of the AI ​​model, in some embodiments, may include a perceptron neural network, a feedforward neural network, a multilayer perceptron neural network, a radial basis function neural network, a recurrent neural network, a long-short-term memory neural network, a sequence-to-sequence neural network model, a modular neural network, a graph-based convolutional neural network, an instance-based model, a feature attribute model, etc.

[0522] In a non-limiting example, an AI model of the claimed invention can include a perceptron neural network configured to receive chemical data as input, perform one or more functions on the input, multiply the output of the one or more functions by a number of weights, and generate a final output that includes one or more candidate compounds. In another non-limiting example, an AI model of the claimed invention can include a multi-layer perceptron neural network including multiple layers, each configured to perform the processes of a single perceptron network, and each layer linked to input and output feeds such that the output of one layer is the input of a subsequent layer. The input to the first layer can be chemical data, and the output of the last layer can be one or more candidate compounds.

[0523] In another non-limiting example, the AI ​​model may include a graph-based convolutional neural network including multiple layers. Each layer of the multiple layers may be linked to input and output feeds such that the output of one layer is the input for the subsequent layer. The input to the first layer may be chemical data, and the output of the last layer may be one or more candidate compounds. The chemical data may be transformed into one or more graph structures before being sent to the multiple layers. In some embodiments, the multiple layers and functions therein may be configured to extract one or more subgraphs from an input graph containing one or more associated node features, and apply one or more attention-based functions to the subgraphs and associated node features to assign importance to particular molecules, bonds, atoms, etc., such that the model can utilize the most relevant patterns determined by the one or more attention-based functions to generate and / or predict new candidate compounds based on inputs including RGN6024. Determining which patterns are relevant and important for discovery may be a component of a training dataset.

[0524] The one or more candidate compounds may include one or more compounds linked to RGN6024, one or more compounds reconstructed from components of RGN6024, one or more compounds derived from RGN6024 by chemical reaction, one or more compounds related to RGN6024, or a combination thereof.

[0525] With respect to methods for implementing AI to discover compounds related to RGN6024 and AI-based systems thereof, the specifications of U.S. Patent No. 10,776,712, issued September 15, 2020, and U.S. Patent No. 11,462,304, issued October 4, 2022, are incorporated herein by reference in their entireties.

[0526] Example 28

[0527] In another exemplary method, the use of one or more of the compounds disclosed herein as drug screening tools for use in in vitro, ex vivo, and in vivo assays has been further characterized as illustrated and described herein below.

[0528] The present invention features a method for screening one or more potential drug candidate compounds to determine whether they are therapeutically effective using any one of the compounds disclosed herein as a reference compound. The method includes: (a) identifying one or more in vitro, ex vivo, or in vivo models for the experiment; (b) contacting at least one cell, body fluid, tissue, organ, or animal with at least one of the reference compound, the potential drug candidate compound, or a negative control compound; (c) determining, in parallel, one or more drug parameters or characteristics from contacting the cell, body fluid, tissue, organ, or animal with at least one of the reference compound, the potential drug candidate compound, or the negative control compound; and (d) comparing the one or more drug parameters or characteristics with those of the negative control, the reference compound, or the drug candidate compound to determine whether the drug candidate compound is therapeutically effective.

[0529] In non-limiting examples, in vitro models may include cell viability assays, cell cycle analysis, immunofluorescence staining assays, immunofluorescence staining of cellular beta-tubulin, tubulin polymerization assays, competitive binding assays, colchicine competitive binding assays, binding site cellular assays, colchicine binding site cellular assays, reversibility assays, kinetic solubility assays, microsomal stability assays, plasma protein binding studies, cell permeability studies, or flow cytometric cell cycle analysis. In another non-limiting example, an ex vivo model includes a resected tumor in a chicken egg model. In another non-limiting example, an in vivo model includes a pharmacokinetic (PK) study, a brain pharmacokinetic (PK) study, a maximum tolerated dose study, an efficacy study, metabolite profiling in hepatocytes, a survivability study, or a tumor size study.

[0530] In some non-limiting examples, the drug parameters or characteristics analyzed include pharmacokinetics (PK), pharmacodynamics (PD), brain pharmacokinetics (brain PK), target site binding, cell viability, cell permeability, blood-brain barrier permeability, efficacy, toxicity and safety, stability, microsomal stability, kinetic solubility, plasma protein binding, microsomal stability, or effects on the cell cycle.

[0531] In some non-limiting examples, cells can be contacted with a reference compound, a potential drug candidate, or a negative control compound. The cells can include primary tumor cells, cancer cells, U87 glioblastoma cells, LN-18 glioblastoma cells, HMC3 microglial cells, patient-derived GBM cells, MDR1-MDCK (multidrug resistance-1-Mandin-Darby canine kidney) cells, human cryopreserved hepatocytes, or cells used in patient-derived xenograft models. In other non-limiting examples, body fluids can be contacted with a reference compound, a potential drug candidate, or a negative control compound. Body fluids can include plasma, cerebrospinal fluid, brain homogenate, urine, whole blood, serum, or tumor homogenate. In some non-limiting examples, tissues can be contacted with a reference compound, a potential drug candidate, or a negative control compound. The tissues can include brain tumor tissue, normal brain tissue, tumor tissue, or solid tumor tissue. In other non-limiting examples, organs can be contacted with a reference compound, a potential drug candidate, or a negative control compound. The organ may include the brain, breast, skin, lung, liver, bone, or connective tissue. In another non-limiting example, the animal may be contacted with a reference compound, a potential drug candidate compound, or a negative control compound. The animal may include a mouse, rat, dog, monkey, rabbit, or pig.

[0532] Example 29

[0533] The following are non-limiting examples of the present invention. It should be understood that the examples are not intended to limit the invention in any way. Equivalents or substitutes are within the scope of the present invention.

[0534] Example 29.1 Oral administration to treat brain cancer (glioblastoma) (relevant example: oral administration in combination with one of the standard treatment options for treating cancer (i.e., chemotherapy and / or temazolamide)).

[0535] A 60-year-old man visits his physician suffering from weakness and numbness on the right side of his body, loss of balance, lethargy, and persistent nausea and vomiting. Sensing the urgency of the man's condition, the physician takes a thorough medical history, performs a neurological examination, and orders a brain MRI with contrast to determine the underlying cause of the man's symptoms. The man's test results reveal that he has brain cancer suspected to be glioblastoma. A brain biopsy confirms the diagnosis. The physician prescribes an oral composition containing a 1 mg / kg dose of the RGN6024 compound described herein in tablet form to be taken once daily to treat the brain cancer. The man adheres to the prescribed regimen, and his symptoms gradually ease with each dose over the next month. The patient experiences mild nausea that is well controlled with anti-nausea medication. A follow-up MRI after four weeks of treatment shows an overall reduction in the size of the glioblastoma.

[0536] Example 29.2 Subcutaneous administration to treat brain cancer (glioblastoma)

[0537] A 60-year-old man visits his physician suffering from weakness and numbness on the right side of his body, loss of balance, lethargy, and persistent nausea and vomiting. Sensing the urgency of the man's condition, the physician takes a thorough medical history, performs a neurological examination, and orders a brain MRI with contrast to determine the underlying cause of the man's symptoms. The man's test results reveal that he has brain cancer suspected to be glioblastoma. A brain biopsy confirms the diagnosis. The physician prescribes a composition containing a compound described herein to be administered subcutaneously once weekly at a dose of 2 mg / kg in a 5 mL syringe to treat the brain cancer. The man adheres to the prescribed regimen, and his symptoms gradually ease with each administration over the next month. The patient experiences mild nausea, which is well controlled with anti-nausea medication. A follow-up MRI after four weeks of treatment shows an overall reduction in the size of the glioblastoma.

[0538] Example 29.3 Intravenous administration to treat brain cancer (glioblastoma)

[0539] A 60-year-old man visits his physician suffering from weakness and numbness on the right side of his body, loss of balance, lethargy, and persistent nausea and vomiting. Sensing the urgency of the man's condition, the physician takes a thorough medical history, performs a neurological examination, and orders a brain MRI with contrast to determine the underlying cause of the man's symptoms. The man's test results reveal that he has brain cancer suspected to be glioblastoma. A brain biopsy confirms the glioblastoma. The physician prescribes a composition containing a compound described herein at a dose of 5 mg / kg infused over 30 minutes once weekly to treat the brain cancer. The man adheres to the prescribed regimen, and his symptoms gradually ease with each administration over the next month. The patient experiences mild nausea, which is well controlled with anti-nausea medication. A follow-up MRI after four weeks of treatment shows an overall reduction in the size of the glioblastoma.

[0540] Example 29.4 Oral administration to treat non-small cell lung cancer (NSCLC) metastasizing to the brain. [Relevant examples: EGFR-mutated, ROS-1-mutated, K-RAS-mutated lung cancer; oral administration in combination with one of the standard therapeutic options for treating cancer (i.e., immunotherapy, targeted therapy)].

[0541] A 75-year-old man visited his doctor after being diagnosed with non-small cell lung cancer (NSCLC) one year ago and is currently taking crizotinib to treat his NSCLC. The man complained of headaches, minor seizures, memory loss, and morning nausea and vomiting. Sensing the urgency of the man's condition, the doctor took a thorough medical history, performed a neurological examination, and ordered a brain MRI with contrast to determine the underlying cause of the man's symptoms. The man's test results reveal that the NSCLC has metastasized to his brain. The doctor prescribes an oral pill containing a 0.5 mg / kg dose of the RGN6024 compound described herein to be administered simultaneously with his crizotinib dose to treat the brain metastases. The man complies with the prescribed regimen and gradually experiences symptomatic relief with each dose. The patient experiences mild nausea that is well controlled with anti-nausea medication.

[0542] Example 29.5 Oral administration to treat triple-negative, Her2-positive breast cancer that has metastasized to the brain [Related example: hormone receptor-positive breast cancer that has metastasized to the brain]

[0543] A 53-year-old woman visits her physician after being diagnosed with breast cancer six months prior. The woman complains of a dull, persistent tension-type headache, mild slurred speech, and sudden mood swings. Sensing the urgency of the woman's condition, the physician takes a th...

Claims

1. A compound according to formula (ID), 【Chemistry 1】 In the formula, R ５ but, 【Chemistry 2】 【Transformation 3】 is selected from R ６ is aryl or heteroaryl, and the aryl and heteroaryl are each independently unsubstituted or each independently substituted with one or more of alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfo, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, or heterocyclic alkylthio.

2. R ６ 2. The compound of claim 1, wherein: 【Chemistry 4】 【Transformation 5】

3. 10. The compound of claim 1, wherein the compound is configured to cross the blood-brain barrier (BBB) ​​of a human or non-human subject.

4. 4. The compound of claim 3, wherein the compound has a central nervous system multiparameter optimization (CNS MPO) score of 4.0 or greater, a Papp score of greater than 10, or an efflux ratio of the compound of less than 2.

0.

5. 10. The compound of claim 1, wherein the compound is effective in treating a health condition, treating cancer or metastasis, preventing cancer cell division, inhibiting tubulin polymerization, destabilizing microtubules, arresting cell division in the G2 / M phase, being cytotoxic to multiple cancer cell lines, targeting blood vessels and vasculature in cancer or tumors, treating a non-neoplastic condition, treating gout, treating familial Mediterranean fever, treating onychomycosis, targeting blood vessels or vasculature, or any combination thereof.

6. 10. A composition comprising at least one compound of claim 1 and one or more of a tag, an inert moiety, or a targeting moiety linked to said compound.

7. 7. The compound of claim 6, wherein the tag is a fluorescent tag, a radioactive tag, biotin, or any combination thereof.

8. 7. The compound of claim 6, wherein the inert moiety is an ester, a carbamate, an aminoacyl ester, or any combination thereof.

9. 8. The compound of claim 7, wherein the targeting moiety is an antibody, a polyethylene glycol (PEG) conjugate, or a long chain polymer, a peptide sequence, or any combination thereof.

10. The compound is 【Transformation 6】 2. The compound of claim 1, wherein:

11. A composition comprising at least one compound according to claim 1.

12. A compound according to formula II-D, 【Transformation 7】 wherein J is 【Transformation 8】 【Chemistry 9】 and L 【Chemistry 10】 【Chemistry 11】 Compounds wherein K is aryl or heteroaryl, and the aryl and heteroaryl are each independently unsubstituted or substituted with one or more groups selected from alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfo, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heterocyclyl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, and heterocyclic alkylthio.

13. 13. The compound of claim 12, wherein K is selected from the following: 【Chemistry 12-1】 【Chemistry 12-2】 【Chemistry 13】

14. 13. The compound of claim 12, wherein the compound is configured to cross the blood-brain barrier (BBB) ​​of a human or non-human subject.

15. 15. The compound of claim 14, wherein the compound has a central nervous system multiparameter optimization (CNS MPO) score of 4.0 or greater, a Papp score of greater than 10, or an efflux ratio of the compound of less than 2.

0.

16. A composition comprising at least one compound according to claim 12.

17. A compound or derivative thereof according to one of the following: 【Chemistry 14-1】 【Chemistry 14-2】 【Chemistry 14-3】 【Chemistry 15】

18. 18. The compound of claim 17, wherein the compound is configured to cross the blood-brain barrier (BBB) ​​of a human or non-human subject.

19. 20. The compound of claim 18, wherein the compound has a central nervous system multiparameter optimization (CNS MPO) score of 4.0 or greater, a Papp score of greater than 10, or an efflux ratio of the compound of less than 2.

0.

20. 18. A composition comprising at least one compound according to claim 17.

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