Compositions and methods for producing 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 of related applications This application is a non-provisional patent application and claims the benefits of U.S. Provisional Patent Application No. 63 / 555,767 filed on 20 February 2024, U.S. Provisional Patent Application No. 63 / 591,709 filed on 19 October 2023, and U.S. Provisional Patent Application No. 63 / 448,964 filed on 28 February 2023, the entirety of which their specifications are incorporated herein by reference.

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

[0003] Despite advances in oncological treatment, cancer remains a leading cause of death due to its high morbidity and mortality rates. While small molecule therapies can be successful in treating some cancers, many have narrow therapeutic indices, are not highly selective, and cause undesirable drug toxicity in the target population. Insufficient penetration into sanctuary sites (e.g., CNS) necessitates administration of high concentrations of some small molecules, 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 develop novel small molecules for more successful treatments of cancer and other health conditions. [Overview of the Initiative]

[0004] The present invention features compounds, compositions, and methods that enable the creation and use of small molecules for the treatment of health conditions. In some embodiments, the disclosure provides novel compounds, compositions, and methods for treating one or more cancers that metastasize to the brain in a subject requiring such treatment. 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 described in the dependent claims. Embodiments of the present invention may be freely combined with one another if they are not mutually exclusive.

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

[0006] While we do not intend to limit the present invention to any theory or mechanism, the technical features of the present invention are considered to 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 are unable to enter the brain, and others whose success rate in crossing the BBB is far lower than that observed herein. For example, some molecules of this application are designed to bind to small cracks on the surface of tubulin protein, thereby allowing smaller molecules to enter and providing improved brain penetration and avoidance of the efflux pump, which would otherwise remove these molecules from the brain. Furthermore, the present invention is considered to advantageously provide highly effective and potent treatments for cancer, including brain cancer (e.g., treatment-resistant glioblastoma), as well as 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] Appropriate methods and materials for carrying out and / or testing embodiments of the disclosure are described below. Such methods, materials, and examples are illustrative and not intended to limit the scope. Other methods and materials similar or equivalent to those described herein may be used. For example, conventional methods well known in the art relating to this disclosure are described in various general and more specific references.

[0008] In certain embodiments, the Disclosure provides compounds for use in treating, preventing, or improving a health condition in a subject, or for use as an adjunct therapy in treating, reducing the onset of, or improving a health condition in a subject that requires such treatment or improvement.

[0009] In some embodiments, the present invention is expressed by formula (ID): [ka] Characterized by compounds produced by In the formula, R5 is [ka] [ka] Selected from, R6 is an aryl or heteroaryl, and each aryl and heteroaryl may be independently unsubstituted or independently 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 compound is configured to cross the blood-brain barrier (BBB) ​​in a human or non-human subject. In certain embodiments, the present invention has a central nervous system multi-parameter optimization (CNS MPO) score of 4.0 or higher, a Papp score greater than 10, or an efflux ratio of less than 2.0 for the compound.

[0012] In certain embodiments, the compound is effective for treating health conditions, cancer or metastasis, preventing cancer cell division, inhibiting tubulin polymerization, destabilizing microtubules, suppressing G2 / M phase cell division, cytotoxicity against multiple cancer cell lines, targeting vascular and vascular structures of cancer or tumors, treating non-neoplastic conditions, treating gout, treating familial Mediterranean fever, treating onychomycosis, targeting vascular or vascular structures, or any combination thereof.

[0013] In certain embodiments, the present invention features a composition comprising a compound according to formula (ID) and one or more tags, inert moieties, or targeting moieties 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] That is the case.

[0015] In some embodiments, the present invention is represented by formula II-D: [ka] Characterized by compounds produced by In the formula, J is [ka] [ka] And, L is [ka] [ka] K is an aryl or heteroaryl, and each aryl and heteroaryl may independently be unsubstituted or independently 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, where K is [ka] [ka] [ka] Selected from.

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

[0018] In some embodiments, the present invention is characterized by a composition comprising a compound according to formula (ID) disclosed herein. In other embodiments, the present invention is characterized by a composition comprising a compound according to formula (II-D) disclosed herein.

[0019] In certain embodiments, the present invention is characterized by a compound comprising 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) ​​in human or non-human subjects. In certain embodiments, the present invention has a central nervous system multi-parameter optimization (CNS MPO) score of 4.0 or higher, a Papp score greater than 10, or an efflux ratio of less than 2.0 for the compound.

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

[0022] Treatment method

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

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

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

[0026] In some embodiments, the health condition includes 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, hematological cancer, gastrointestinal cancer, medulloblastoma, or other solid organ cancer, cell carcinoma or tissue carcinoma, 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 includes brain cancer or cancer that can metastasize to the brain. In some embodiments, the cancer may be glioblastoma, high-grade glioma, other brain cancers, noncellular lung cancer (NSCLC) before or after metastasis to the brain, angiogenic carcinoma, or any combination thereof.

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

[0028] In some embodiments, the method further comprises the administration of one or more antimicrobial agents, chemotherapeutic agents, other anticancer therapies, or antibodies or fragments thereof. In some embodiments, the antimicrobial agent comprises one or more antiviral, bactericidal, antifungal, or antibacterial agents, or other antimicrobial agents. In further embodiments, the antimicrobial agent may be an antibacterial agent (antibiotic) such as doxycycline or tetracycline, or other antibiotics, such as generally applicable antibiotics.

[0029] In some embodiments, the chemotherapeutic agent includes one or more of the following: temozolomide, lomustine, verzutifan, 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 anticancer therapy comprises one or more of chemotherapy, radiotherapy, immunotherapy, and / or surgery. In other embodiments, the anticancer therapy further comprises one or more anticancer therapies or treatments, e.g., one or more chemotherapeutic agents, radiotherapy, small molecules, and immunomodulators. In some embodiments, the anticancer therapy or treatment may be administered separately from the compounds or derivatives disclosed herein.

[0031] In other embodiments, antimicrobial agents, chemotherapeutic agents, other anticancer therapies, or antibodies or fragments thereof are administered before, during, or after administration of one or more compounds or compositions or derivatives thereof disclosed herein.

[0032] In some embodiments, the health condition is cancer, and one or more anticancer treatments include one or more administrations of temozolomide, lomustine, verzutifan, or any combination thereof, before, during, or after administration of the composition. In other embodiments, the health condition is NSCLC, and one or more anticancer treatments include one or more administrations 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 the division of cancer cells. 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 dose 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 to 5 mg / kg twice daily. In other embodiments, the therapeutically effective dose in mice is in the range of 1 to 25 mg / kg.

[0035] In some embodiments, the route of administration is one or more of the following: intravenous injection, oral administration, subcutaneous injection, intramuscular injection, intrasternal injection, intrathecal administration, intratumoral, intravascular, intracerebral injection, intracisional, intraventricular, intranasal or inhalation, parenteral, buccal, enteral, intraperitoneal, inhalable, infusion, intramuscular, ocular, intravitreous, ear, rectal, sublingual, topical, percutaneous, intrapulmonary, intrauterine, vaginal, via ultrasound-mediated blood-brain barrier disruption, implantable devices, injection techniques, or nanoparticle-based delivery.

[0036] In some embodiments, the composition is in the form of tablets, pills, coated tablets, or coated pills. In some embodiments, the effective dose is administered to the subject before, during, or after administration of at least one compound at least once daily, 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 preferred 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 may further include administering a therapeutically effective amount of one or more anticancer agents or treatments to the subject before, concurrently with, or after administering at least one effective amount of a compound or formulation disclosed herein to the subject. According to these embodiments, one or more anticancer agent treatments may include radiotherapy. In some embodiments, the cancer may be brain cancer, and the one or more anticancer agents include administering one or more of temozolomide, lomustine, verzutifan, or any combination thereof. In other embodiments, the cancer may be NSCLC, and the one or more anticancer treatments include administering one or more of crizotinib, osimertinib, or any combination thereof to the subject, in addition to the compounds or compound-containing formulations disclosed herein.

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

[0040] How to use

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

[0042] In some embodiments, the present invention relates to a drug screening method for identifying therapeutically effective drug candidates for treating a health condition in a subject requiring such treatment, the method comprising: (a) identifying an in vitro or in vivo model of the health condition; (b) administering the 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 therapeutically effective drug candidates.

[0043] In some embodiments, the present invention relates to a method for detecting target cells having abnormal cell division, the method comprising: (a) identifying a sample containing target cells having abnormal cell division; (b) performing an affinity-based assay using a small molecule conjugated with a tag to contact the target cells having abnormal cell division with the small molecule conjugated with a tag; and (c) determining whether the small molecule conjugated with a tag binds to the target cells 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 immuno-based assay, receptor-based assay, antibody-based assay, nanoparticle-based assay, chemical assay, optical assay, or 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, affinity-based assays are colchicine competitive binding assays.

[0044] In some embodiments, the present invention relates to a method for detecting a protein expressed by target cells having abnormal cell division, the method comprising: (a) identifying a sample containing a protein expressed by target cells having abnormal cell division; (b) performing an affinity-based assay using a small molecule conjugated with a tag to contact the protein expressed by target cells having abnormal cell division with the small molecule conjugated with a tag; and (c) determining whether the small molecule conjugated with a tag binds to the protein expressed by target cells 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 immuno-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, affinity-based assays include gel electrophoresis, enzyme-linked immunosorbent assays, immunoblot assays, fluorescence intensity assays, fluorescence anisotropy assays, fluorescence energy transfer assays, surface plasmon resonance (SPR) assays, light scattering assays, forward binding assays, dissociation assays, or reverse binding assays. In some embodiments, affinity-based assays include colchicine competitive binding assays.

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

[0046] In other embodiments, a method for detecting or isolating 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 pull-down assay using a small molecule conjugated with a tag to contact the protein expressed by the target cell having abnormal cell division with the small molecule conjugated with a tag; and (c) selectively isolating the protein expressed by the target cell having abnormal cell division. In other embodiments, 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 the treatment of cancer, comprising (a) one of the compositions or derivatives thereof disclosed herein in a therapeutically effective dose, (b) at least one container for storing the composition, and (c) dosage and administration instructions.

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

[0050] Any feature or combination of features described herein is included within the scope of the invention, provided that the features included in any such combination are not inconsistent with each other, as is evident from the context, this specification, and the knowledge of those skilled in the art. Additional advantages and aspects of the invention are evident in the following detailed description and claims.

[0051] The following drawings form part of this specification and are included to further demonstrate specific embodiments of the present disclosure. Specific embodiments can be better understood by referring to one or more of these drawings in combination with the detailed description of the specific embodiments presented herein. [Brief explanation of the drawing]

[0052] [Figure 1] Exemplary experiments illustrating the chemical reaction equation for synthesizing 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidine-5-yl)oxy)phenyl)cyclobutan-1-carboxamide (compound B19) according to specific embodiments of the present disclosure are shown.

[0053] [Figure 2] Exemplary experiments illustrating the chemical reaction equation for synthesizing 3-methoxy-N-(4-((2-(4-methoxypiperidine-1-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)cyclobutan-1-carboxamide (compound B46) according to a particular embodiment of the present disclosure are shown.

[0054] [Figure 3] Exemplary experiments illustrating the chemical reaction equation for synthesizing 3-methoxy-N-(5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide (compound B120) according to specific embodiments of the present disclosure are shown.

[0055] [Figure 4] Exemplary experiments illustrating the chemical reaction equation for synthesizing 3-methoxy-N-(5-(4-morpholinophenoxy)thiazole-2-yl)cyclobutan-1-carboxamide (compound B121) according to specific embodiments of the present disclosure are shown.

[0056] [Figure 5]Exemplary experiments illustrating the chemical reaction equation for synthesizing N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxycyclobutane-1-carboxamide (compound B72) according to specific embodiments of the present disclosure are shown.

[0057] [Figure 6] Exemplary experiments illustrating the chemical reaction equation for synthesizing 3-methoxy-N-(3-methyl-4-((2-morpholinothiazole-5-yl)oxy)phenyl)cyclobutane-carboxamide (compound B71) according to specific embodiments of the present disclosure are shown.

[0058] [Figure 7] An exemplary experiment illustrating the chemical reaction equation for synthesizing compound 126:3-methoxy-N-(5-((6-morpholinopyridine-3-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide (compound B126) according to a particular embodiment of the present disclosure is shown.

[0059] [Figure 8] Exemplary experiments illustrating the chemical reaction equation for synthesizing 3-methoxy-N-(5-((2-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide (compound B122) according to specific embodiments of the present disclosure are shown.

[0060] [Figure 9] Exemplary experiments illustrating the chemical reaction equation for synthesizing 3-methoxy-N-(5-(4-(piperidine-1-yl)phenoxy)thiazole-2-yl)cyclobutan-1-carboxamide (compound B125) according to specific embodiments of the present disclosure are shown.

[0061] [Figure 10]Exemplary experiments illustrating the chemical reaction equation for synthesizing N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxycyclobutan-1-carboxamide (compound B130) according to specific embodiments of the present disclosure are shown.

[0062] [Figure 11] Exemplary experiments illustrating the chemical reaction equation for synthesizing 3-methoxy-N-(5-((2-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide (compound B140) according to a particular embodiment of the present disclosure are shown.

[0063] [Figure 12A] Exemplary experiments illustrating the chemical reaction equation for synthesizing 3-methoxy-N-(5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide (RGN6024) according to specific embodiments of the present disclosure are shown.

[0064] [Figure 12B] The proposed metabolic pathways of RGN6024 in mouse, rat, dog, monkey, and human hepatocytes are shown.

[0065] [Figure 13] Exemplary experiments illustrating the chemical reaction equation for synthesizing N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide (compound B138) according to specific embodiments of the present disclosure are shown.

[0066] [Figure 14]An exemplary experiment illustrating the chemical reaction equation for synthesizing N-(5-((2-(2-oxa-7-azaspiro[4.4]nonane-7-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide (compound B137) according to a particular embodiment of the present disclosure is shown.

[0067] [Figure 15] Exemplary experiments illustrating the chemical reaction equation for synthesizing 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidine-5-yl)oxy)phenyl)bicyclo[1.1.1]pentan-1-carboxamide (compound B89) according to specific embodiments of the present disclosure are shown.

[0068] [Figure 16] Exemplary experiments illustrating the chemical reaction equation for synthesizing 3-methoxy-N-(5-(4-morpholinophenoxy)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide (compound B155) according to specific embodiments of the present disclosure are shown.

[0069] [Figure 17] Exemplary experiments illustrating the chemical reaction equation for synthesizing N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide (compound B118) according to specific embodiments of the present disclosure are shown.

[0070] [Figure 18] Exemplary experiments illustrating the chemical reaction equation for synthesizing N-(4-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide (compound B117) according to specific embodiments of the present disclosure are shown.

[0071] [Figure 19A]An exemplary experiment demonstrating inhibition of tubulin polymerization is shown. The effects of compounds B130, B144, and B147 at 5 μM concentrations on tubulin polymerization are demonstrated. Colchicine is used as a positive control.

[0072] [Figure 19B] This exhibits an exemplary experiment demonstrating the inhibition of tubulin polymerization. The effect of 5 μM RGN6024 on tubulin polymerization is shown. Colchicine (COL) is the positive control, and PX is paclitaxel.

[0073] [Figure 20A] An exemplary experiment demonstrating the inhibition of tubulin polymerization is shown. The effects of compound B138 at concentrations of 0.1, 0.3, 1, and 3 μM are shown.

[0074] [Figure 20B] An exemplary experiment demonstrating the inhibition of tubulin polymerization is shown. The effects of RGN6024 at concentrations of 0.3, 1, 3, and 5 μM are demonstrated.

[0075] [Figure 21] A fluorescence-based colchicine competitive binding assay using 50 μM RGN6024 is shown according to a specific embodiment of this disclosure. Nocodazole is used as a positive control.

[0076] [Figure 22] This disclosure presents an N,N'-ethylene-bis(iodoacetamide) (EBI) competitive assay using compound B138 in the MCF7 cell line, demonstrating compound binding at the colchicine binding site according to a specific embodiment of this disclosure. Colchicine is the positive control, and vinblastine is the negative control. 3071 is the internal control compound.

[0077] [Figure 23] This document illustrates an exemplary experiment demonstrating the concentration of compound B138 in plasma after oral administration of 30 mg / kg to a rodent model, according to a specific embodiment of the present disclosure.

[0078] [Figure 24] This document illustrates an exemplary experiment showing the concentration of RGN6024 in plasma after oral administration of 30 mg / kg to a rodent model, according to a specific embodiment of the present disclosure.

[0079] [Figure 25] This document illustrates exemplary experiments showing the concentrations of RGN6024 in plasma and brain tissue after oral administration of 30 mg / kg to a rodent model, according to specific embodiments of this disclosure.

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

[0081] [Figure 26B] This shows the amount of compound B155 in plasma, brain, and tumors at the end of the efficacy study.

[0082] [Figure 27] This demonstrates the visibly superior effect of RGN6024 on resistant tumors. Figure 27 shows tumors excised from an animal model, comparing placebo, standard treatment temozolomide (TMZ), and RGN6024, illustrating the effect of RGN6024 on tumor growth and size.

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

[0084] [Figure 29]This study demonstrates that RGN6024 reduces tumor growth by 98% in an intracranial model of glioblastoma. BT-142 tumors (glioblastoma) were transplanted into the brain. RGN6024 versus placebo was administered orally at 15 mg / kg twice daily. The observed bioluminescence signal was approximately proportional to tumor size. Treated cells showed only 2% of the bioluminescence signal observed in untreated cells.

[0085] [Figure 30] This study demonstrates body weight as a proxy for health in an intracranial model of glioblastoma. Animals treated with RGN6024 maintained their body weight compared to placebo-treated animals, which showed significant weight loss and increased tumor signaling. The placebo group showed increased weight loss between days 18 and 21 compared to treatment.

[0086] [Figure 31] This study demonstrates that RGN6024 induces G2 / M cell cycle arrest compared to a control (vehicle only) and sabizabrin (a clinical-stage tubulin-targeted therapy with minimal brain penetration). RGN6024 inhibits the division of brain cancer cells, causing them to quiesce in the G2 / M phase. The literature indicates that G2 / M phase cells are more sensitive to radiotherapy.

[0087] [Figure 32] This paper presents predictions for combination therapy with RGN6024. Literature indicates that radiotherapy targets cancer cells in the G2 / M phase of cell division, which are more sensitive to radiotherapy. Preliminary results suggest an additive effect between radiotherapy and RGN6024.

[0088] [Figure 33]This study demonstrates that RGN6024 exhibits dose-dependent inhibition of tubulin polymerization in biophysical assays. The dose-dependent effect of RGN6024 on the target shows that (1) increasing dose leads to increased inhibition of tubulin polymerization, consistent with binding to tubulin; and (2) RGN6024 (50 μM) achieves the same level of tubulin polymerization inhibition as colchicine (positive control, 5 μM).

[0089] [Figure 34] This demonstrates that RGN6024 binds to the colchicine binding site. Fluorescence is generated by colchicine binding within the colchicine pocket. RGN6024 competes with colchicine and its binding pocket, as indicated by the decrease in fluorescence. This is consistent with RGN6024 targeting the colchicine binding site.

[0090] [Figure 35] This shows Western blot target site engagement in living cells. RGN6024 inhibits modification by binding to 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 that β-tubulin is modified in the absence of the drug. The EBI+6024 lane shows that RGN6024 reduces β-tubulin modification. The EBI+colchicine (positive control) lane shows that β-tubulin is protected from modification.

[0091] [Figure 36] RGN6024 (5 μM concentration) demonstrates cleanliness against 44 drug targets with the most severe adverse drug reactions. Inhibition is less than 30% for all targets. RGN6024 is suitable for preclinical trials.

[0092] [Figure 37]The efficacy of RGN6024 is demonstrated in cancer cell lines (high-grade glioma cell lines: BT142, LN-18, U-87, LN-229, U118, and T98G; cell lines prone to brain metastasis: SKMEL5, HCC1806, and NCI-H460).

[0093] [Figure 38] This study demonstrates that RGN6024 is orally available to mice at a well-tolerated dose. [Figure 38] The plasma concentrations (ng / mL) of IV (3 mg / kg) versus PO (30 mg / kg) are shown over time (8 hours). Half-life (T1 / 2) (h) = 2.64, peak concentration (CMAX) (ng / mL) = 7020. Oral bioavailability exceeds 50%, and CMAX is close to approximately 17 μM.

[0094] [Figure 39] This report presents a 5-day maximum tolerable dose study in which untreated mice were observed for 5 days in comparison to mice treated with 10 mg / kg, 30 mg / kg, or 100 mg / kg of RGN6024. The maximum oral tolerable dose of RGN6024 is 30 mg / kg to 100 mg / kg. RGN6024 is water-soluble in 30% 2-hydroxypropyl-β-cyclodextrin in physiological saline.

[0095] [Figure 40] This paper presents the physicochemical and ADME properties of RGN6024, including important brain permeability characteristics (molecular weight, LogD, MPO score), important oral administration characteristics (kinetic solubility), and cell permeability (MDR1-MDCK cells; Papp and efflux ratio). RGN6024 exhibits an MPO score > 4.0, which predicts good brain permeability, is soluble in saline formulations, has significant permeability (Papp > 10) resulting in better diffusion, and has an efflux ratio < 2.0, suggesting it is not an MDR1 pump substrate.

[0096] [Figure 41]This demonstrates the in vitro ADME properties of RGN6024. RGN6024 exhibits low clearance and a high half-life, which suggests the possibility of good plasma levels in humans, moderate protein binding in humans, a higher percentage of free drug than in mouse data, and no significant inhibition of the cytochrome P450 isoforms tested.

[0097] [Figure 42] It demonstrates the maximum brain penetration of tubulin-targeted therapies. RGN6024 has 14 times more drug in the brain compared to tubulin-targeted therapies ANG1005, sabizabrin, eribulin, paclitaxel, and unesbrin.

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

[0099] [Figure 44] 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 far 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 sabizabrin. While colchicine and sabizabrin have similar affinities to tubulin protein, their safety profiles differ significantly.

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

[0102] Figures 47A–47C demonstrate the reversibility of RGN6024 binding to tubulin protein compared to colchicine (high toxicity) and sabizabrin (low toxicity), as evidenced by the increased cell viability after washout. [Figure 47A] This shows that there is little increase in cell viability after colchicine washout. [Figure 47B] This shows an increase in cell viability after sabizabrin washout. [Figure 47C] This shows increased cell viability after RGN6024 washout. The reversibility of RGN6024 binding to tubulin suggests improved safety in humans, similar to the reversibility of sabizabrin binding to tubulin, which is consistent with low toxicity in humans.

[0103] [Figure 48] The image shows 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 body weight of mice over the first few days after the start of treatment with RGN6024. It also shows that vehicle (untreated) mice lose weight due to tumor burden in the brain. Mice treated with RGN6024 show less weight loss compared to vehicle mice.

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

[0106] [Figure 51] This report demonstrates that RGN6024 can be used to treat health conditions such as cancer by targeting cell cycle elements and can induce 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 a compound for 24 hours and then fixed with 4% paraformaldehyde for 20 minutes. After washing the cells with PBS, they were permeabilized with FoxP3 perm buffer (BD Biosciences) at room temperature for 10 minutes. 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 magnification) were obtained using an automated high-content imaging microscope (Operetta, Perkin Elmer).

[0107] [Figure 52] Current FDA-approved therapies related to tubulin interaction show that they bind to tubulin at large cracks on the protein surface, are too large for efficient brain penetration, are substrates for the efflux pump, and therefore do not penetrate the brain. RGN6024 is smaller than the FDA-approved therapies, can pass through the blood-brain barrier passively, and does not act as an efflux pump substrate.

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

[0109] [Figure 54] This report describes the testing of RGN6024 in various high-grade glioma strains, as well as in breast, lung, and melanoma tumors with a tendency to metastasize to the brain. A median potency of 91 nM was observed. In comparison, the brain penetration level of RGN6024 is 25 times higher than the concentration required to see an effective response in these in vitro cancer models. [Modes for carrying out the invention]

[0110] term

[0111] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which the disclosed inventions belong. The singular terms “a,” “an,” and “the” include plural referents unless otherwise explicitly indicated by the context. Similarly, the word “or” is intended to include “and” unless the context clearly indicates otherwise.

[0112] The term "includes" means that other elements may be present in addition to the defined elements presented. The use of "includes" indicates inclusion, not limitation. In other words, the term "includes" means "primarily includes, but not necessarily alone." Furthermore, variations of the word "includes," such as "comprise" and "comprises," have the same meaning. In one view, the techniques described herein relate to the compositions, systems, methods, and their respective constituent components described herein as essential to the invention, but there is still room to include unspecified elements, whether essential or not ("includes").

[0113] To facilitate the consideration of various embodiments of this disclosure, the following explanations of specific terms are provided.

[0114] As used herein, the term “approximately” may mean ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1% of the listed values, such as volume, dosage, temperature, time, or percentage.

[0115] As used herein, the terms "B137" or "Compound B137" or "C21" or "Compound C21" refer to compounds comprising the following formula: N-(5-((2-(2-oxa-7-azaspiro[4.4]nonane-7-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide, with structure: [ka] It can be used interchangeably to refer to compounds having [this characteristic].

[0116] As used herein, the terms "B138" or "Compound B138" or "C18" or "Compound C18" refer to a compound comprising the following formula: N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide, with structure: [ka] It can be used interchangeably to refer to compounds having [this characteristic].

[0117] As used herein, the terms “RGN6024” or “Compound B139” or “B139” refer to a compound comprising the following formula: 3-methoxy-N-(5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide, with structure: [ka] It can be used interchangeably to refer to compounds having [this characteristic].

[0118] As used herein, the terms "B141" or "Compound B141" or "C2" or "Compound C2" refer to a compound of the formula: 3-methoxy-3-methyl-N-(5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide, and structure: [ka] It can be used interchangeably to refer to compounds having [this characteristic].

[0119] As used herein, the terms "B142" or "Compound B142" or "C7" or "Compound C7" refer to a compound of the formula: 3-(cyclopropylmethoxy)-N-(5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide, and structure: [ka] It can be used interchangeably to refer to compounds having [this characteristic].

[0120] As used herein, the terms "B144" or "Compound B144" or "C6" or "Compound C6" refer to compounds of the formula: 3-isopropoxy-N-(5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide, and structure: [ka] It can be used interchangeably to refer to compounds having [this characteristic].

[0121] As used herein, the terms "B146" or "Compound 146" or "C3" or "Compound C3" refer to a compound of the formula: 3-methoxy-1-methyl-N-(5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide, structure: [ka] It can be used interchangeably to refer to compounds having [this characteristic].

[0122] As used herein, the terms "B155" or "Compound 155" or "C27" or "Compound C27" or "C89" or "Compound C89" refer to compounds of the formula: 3-methoxy-N-(5-(4-morpholinophenoxy)thiazole-2-yl)bicycl[1.1.1]pentan-1-carboxamide, and structure: [ka] It can be used interchangeably to refer to compounds having [this characteristic].

[0123] As used herein, the terms "B156" or "Compound 156" or "C14" or "Compound C14" refer to compounds of the formula: 2-methoxy-N-(5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)cyclopropane-1-carboxamide, and structure: [ka] It can be used interchangeably to refer to compounds having [this characteristic].

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

[0125] As used herein, the term "isomer" refers to compounds that have the same number and type of atoms, and therefore the same molecular weight, but differ in terms of the structural arrangement or composition of their atoms.

[0126] Unless otherwise stated, the structures shown herein also include all isomers of said structures (e.g., enantiomers, diastereomers, and geometric (or conformational) forms), such as the R and S configurations of each chiral center, the Z double bond isomer and the E double bond isomer, and the Z conformational isomer and the E conformational isomer. Thus, single stereochemical isomers of the compound, as well as enantiomer mixtures, diastereomer mixtures, and geometric (or conformational) mixtures, are within the scope of the invention. Unless otherwise stated, all tautomeric forms of the compound of the invention are within the scope of the invention. Unless otherwise stated, the structures shown herein also include compounds that differ only in the presence of one or more isotopically enriched atoms, for example, the substitution of hydrogen with deuterium or tritium, or 13 C concentrated carbon or 14 Compounds having this structure, including carbon substitution with 13C-enriched carbon, are within the scope of the present invention. Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents according to the present invention. In certain embodiments, the warhead portion of the provided compound contains one or more deuterium atoms.

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

[0128] The terms “salt” and “pharmaceutically acceptable salt,” as used herein, refer to salts that, within the bounds of sound medical judgment, are suitable for use in contact with human and lower animal tissues without excessive toxicity, irritation, and allergic reactions, that meet a reasonable benefit-risk ratio, and that are effective for their intended use. “Pharmacologically acceptable salt” may refer to salts that may be formed when the compounds herein have an acidic group such as a carboxyl or a basic group such as an amino or imino. In some embodiments, salts of the compounds disclosed herein may be formed with acidic groups and may include, but are not limited to, alkali metal salts such as sodium, potassium, or lithium salts, alkaline earth metal salts such as calcium or magnesium salts, metal salts such as aluminum or iron salts, amine salts such as ammonium salts, and organic salts such as t-octylamine salt, dibenzylamine salt, morpholine salt, glucosamine salt, phenylglycine alkyl ester salt, ethylenediamine salt, N-methylglucamine salt, guanidine salt, diethylamine salt, triethylamine salt, dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt, chloroprocaine salt, procaine salt, diethanolamine salt, N-benzylphenethylamine salt, piperazine salt, tetramethylammonium salt, or tris(hydroxymethyl)aminomethane salt, as well as amino acid salts such as glycine salt, lysine salt, arginine salt, ornithine salt, glutamate, and aspartate.In some embodiments, salt derivatives of the compounds disclosed herein formed with a basic group may include, but are not limited to, hydrogen halides, e.g., hydrofluorides, hydrochlorides, hydrobromids, or hydroiodides; inorganic salts, e.g., nitrates, perchlorates, sulfates, or phosphates; lower alkanesulfonates, e.g., methanesulfonates, trifluoromethanesulfonates, or ethanesulfonates; arylsulfonates, e.g., benzenesulfonates, or p-toluenesulfonates; organic salts, e.g., acetates, malates, fumarates, succinates, citrates, ascorbicates, tartrates, oxalates, or maleates; and amino acid salts, e.g., glycine salts, lysine salts, arginine salts, histidine salts, ornithine salts, glutamates, or aspartates. In certain embodiments, pharmaceutically acceptable salts of the compounds disclosed herein may absorb water upon continued exposure to air or recrystallization to form hydrates for use in the formulations disclosed herein.

[0129] As used herein, the terms “active metabolite” and “metabolite” refer to biomolecules involved in metabolism. The term “metabolite” may also refer to intermediate or final products of metabolic reactions catalyzed by naturally occurring enzymes within cells. While the term “metabolite” is used to describe small molecule compounds, it 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 molecular entities (e.g., atoms, ions, molecules, etc.) that are formed directly or indirectly from reactants that are converted into products in a multi-step chemical reaction. Reactive intermediates can be reactive, short-lived, and high-energy, and typically react further to yield the final product.

[0131] As used herein, the term “tagged compound” refers to a compound that has been 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] As used herein, the term “prodrug” refers to a compound that is made more active in vivo by the metabolism of a precursor drug. The compounds and compositions described herein may 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 readily undergo chemical transformation under physiological conditions to yield an active compound. Furthermore, prodrugs may be converted to an active compound by chemical or biochemical methods in an ex vivo environment.

[0133] As used herein, the term “drug conjugate” refers to the covalent bonding of a drug or prodrug to a natural or synthetic molecular carrier for a specific application. Drug conjugations 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 conjugations 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 consist of a low molecular weight, high-affinity targeted ligand, a linker, and a drug payload.

[0134] As used herein, the term "alkyl" refers to C1-C 20This refers to saturated aliphatic hydrocarbon groups, including linear and branched groups. Typical 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, I-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, and 2,3-dimethylbutyl. This includes, but is not limited to, diethylpentyl, 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 their branched-chain isomers. The alkyl groups may be lower alkyl groups having 1 to 6 carbon atoms. Typical 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, and 2,3-dimethylbutyl.The alkyl group may be substituted or unsubstituted. If substituted, the substituent 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 esters.

[0135] As used herein, the term "alkylsulfo" refers to esters of alkanesulfonic acids. As used herein, the term "alkylamino" refers to alkyl substituents bonded to an amino group. Typical examples include phenylalanino, threonine, tryptophan, tyrosine, barino, and N. 2 -Glutaamino, N 2 - Histidino, N 4 -Asparagus is one example.

[0136] As used herein, the term “cycloalkyl” refers to a saturated and / or partially unsaturated monocyclic or polycyclic hydrocarbon group having 3 to 20 carbon atoms. Typical examples of monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, and cyclooctyl. Polycyclic cycloalkyls may include cycloalkyls having spiro rings, fused rings, and crosslinking rings. Typical 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. Cycloalkyls as used herein may be substituted or unsubstituted. If substituted, the substituent 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 esters. 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 members of that ring. 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 member of that ring. Cycloalkyl, heterocyclic alkyl, and heterocyclic alkylthio as used herein may be substituted or unsubstituted.When substituted, the substituents can be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfone, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, heterocyclic alkylthio, carbonyl, carboxy or carboxylic acid ester.

[0137] As used herein, the term "alkoxyl" refers to an alkyl group singly bonded to oxygen. Representative examples of alkoxyl groups include, but are not limited to, methoxyl, ethoxyl, etc. As used herein, the term "cycloalkoxyl" refers to a cycloalkyl group bonded to oxygen. Representative examples of cycloalkoxyl groups include, but are not limited to, cyclomethoxyl, cycloethoxyl, etc.

[0138] As used herein, the term "heterocyclic alkoxyl" refers to a cycloalkoxyl group having at least two different elemental atoms as members of the ring. The 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, alkylsulfone, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocyclic alkoxyl, cycloalkylthio, heterocyclic alkylthio, carbonyl, carboxy or carboxylic acid ester.

[0139] As used herein, the term "alkenyl" 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, and the like. As used herein, the term "alkynyl" refers to an alkyne, i.e., a fragment formed from a triple bond, by removing one hydrogen atom from any carbon atom.

[0140] As used herein, the term "aryl" refers to an organic group derived from an aromatic ring from which one hydrogen atom has been removed. Representative examples of aryl groups are phenyl, naphthyl, tolyl, xylyl, and the like. As used herein, the term "heteroaryl" refers to a 5- to 14-member aryl having 1 to 4 heteroatoms selected from O, S, and N as ring atoms and the remaining ring atoms being C. Examples of heteroaryl groups are furan, thiophene, pyridine, pyrrole, N-alkylpyrrole, pyrimidine, pyrazine, imidazole, tetrazolyl, and the like. Heteroaryl herein can be condensed with aryl, heterocyclic alkyl, or cycloalkyl, and the ring bonded to the parent structure is heteroaryl. Heteroaryl in this specification can be substituted or unsubstituted. When substituted, the substituent can be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxyl, alkylsulfone, 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 the -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 iodine. As used herein, "thiol" refers to an organosulfur compound in the form of R-SH, where R represents alkyl or other organic substituents. 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] As used herein, the term “heterocyclyl” refers to a monovalent group formed by removing a hydrogen atom from any ring atom of a heterocyclic compound. Typical examples of heterocyclyls include non-aromatic monocyclic, bicyclic, tricyclic, or spirocyclic ring systems containing up to seven atoms in each ring. Heterocyclyls as used herein may be substituted or unsubstituted. If substituted, the substituent 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 esters.

[0143] As used herein, the terms “bicyclic” and “bicyclic ring” refer to a polycyclic molecule characterized by two bonded rings having at least two common atoms. Examples of bicyclic rings include fused bicyclic rings and bridging bicyclic rings. Non-exclusive 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 “bridging bicyclic” or “bridging bicyclic ring” refer to a molecule characterized by two rings bonded together, sharing three or more atoms, with the two bridgeheads separated by a “bridge” containing at least one atom. Bicyclic compounds containing bridges are typically rigid formations 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 may be substituted or unsubstituted. If substituted, the substituent 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 esters.

[0144] As used herein, the term “may be 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 is substituted. It should be understood that the term “substituted” includes the requirement that such substitution conforms to the allowable valencies of the atom being substituted and the substituent, and that the substitution results in a stable compound (i.e., one that does not spontaneously undergo deformation by rearrangement, cyclization, or elimination). In certain embodiments, a single atom may be substituted with two or more substituents, provided that such substitution conforms to the allowable valencies of the atom. Appropriate substituents are defined herein for each substituted or may be substituted group.

[0145] As used herein, the term “health condition” refers to a disease, injury, disability, or physical or mental condition.

[0146] The term "therapeutic dose" can refer to a dose that is sufficient to achieve the desired therapeutic outcome or to have an effect on an undesirable symptom, but is generally insufficient to cause harmful side effects. The specific therapeutic dose level for any particular patient will depend on a variety of factors, including the disorder being treated and its severity; the specific composition used; the patient's age, weight, overall health, sex, and diet; the time of administration; the route of administration; the rate of excretion of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used, as well as similar factors well known in the medical field. For example, it is well within the scope of the art to start with a dose of a compound at a level lower than necessary 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 the purpose of administration. Consequently, a single-dose composition may contain such an amount, or the amount that constitutes a daily dose. The dose can be adjusted by the respective physician in case of any contraindications. The dose may vary and may be administered daily, daily or for several days, weekly, twice a week, etc., in single or multiple doses. Guidance on appropriate dosages for a given class of medications can be found in the literature.

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

[0148] In some cases, the dose may be administered to the subject once daily or in divided doses throughout the day, depending on the subject's clinical response to the drug, determined by methods known in the art. This dose may be administered to the subject once daily, once weekly, or for several days, and then discontinued if the subject responds immediately, or the dose may be administered daily until a clinical response is observed. Those skilled in the art can monitor the subject's clinical response to the administration of the composition and administer additional doses as needed. The composition is intended to be administered to the subject daily, on an alternating daily basis, weekly, or at any interval in between.

[0149] In some embodiments, it is particularly advantageous to formulate the composition into dosage units for ease of administration and dose uniformity. A dosage unit refers to a physically distinct unit suitable as a unit dose for the target to be treated, and each unit contains a predetermined amount of the active compound calculated to produce the desired therapeutic effect.

[0150] The dose may vary within this range depending on the dosage form and route of administration used. Dosages may be formulated in animal models to achieve a concentration range including the IC50 (i.e., the test compound concentration that achieves the suppression of up to half of the symptoms) determined in cell culture. Such information can be used to more accurately determine an effective dose in humans.

[0151] The term "pharmaceutically acceptable" can refer to any dosage form of compound, formulation, or composition that is suitable for contact with human subjects or tissues, and, if necessary, also suitable for use in animals, without excessive toxicity or irritation, and with a reasonable benefit-to-risk ratio that is commensurate with a reduced side effect or complication of its administration as a consumable, and within the bounds of sound medical judgment.

[0152] As used herein, the terms “individual,” “subject,” “host,” “animal,” and “patient” are interchangeable and refer to any subject or any mammal, e.g., human (e.g., adult, adolescent, infant, elderly, child, baby, and fetus), companion animal (e.g., pet, horse), livestock, or other animal, as appropriate, with respect to the diagnosis, treatment, prevention, or therapy.

[0153] As used herein, the terms “to treat,” “treating,” and “treatment” may refer to both therapeutic actions and preventive or preventive measures aimed at preventing, reducing, slowing (mitigating), inhibiting, or eliminating an undesirable physiological change, symptom, disease, or disorder (e.g., cancer).

[0154] For the purposes of the present invention, beneficial or desired clinical outcomes include, but are not limited to, symptom relief, reduction of disease severity, stabilization (i.e., non-exacerbation) of the disease, delay or slowing of disease progression, improvement or mitigation of the disease state, and remission (whether partial or total), whether detectable or undetectable. "Treatment" may also mean extending survival compared to the expected survival time without treatment. Those requiring treatment include those who already have a condition or disability, as well as those who are prone to developing a condition or disability, or for whom a condition or disability should be prevented or its onset should be delayed. In some cases, subjects or patients may be identified (e.g., diagnosed) as having a disease or condition prior to administration of the composition of the present invention. Subjects at risk of disease or disability may be identified, for example, by any or a combination of appropriate diagnostic or prognostic assays known in the art.

[0155] As used herein, the term “administration” means the act of giving 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 may include the subarachnoid space of the brain or spinal cord (subarachnoid space), the eyes (ocular), the mouth (oral), the skin (topical or transdermal), the nose (transnasal), the lungs (inhalation), the oral mucosa (cheek), the ears, the rectum, the vagina, and injections (e.g., intravenous, subcutaneous, intratumoral, intraperitoneal, etc.).

[0156] As used herein, the terms “concurrent administration” or “to administer concurrently” refer 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 this disclosure may be administered alone or concurrently with another compound or standard agents known in the art to a subject. Concurrent administration means administering compounds individually or in combination concurrently or sequentially. Dosage and administration intervals can be individually adjusted to provide effective levels of the compound for the specific clinical indication being treated. This provides a treatment regimen that is appropriate to the severity of the individual’s disease condition.

[0157] In one example, the compositions of the present invention may be administered orally, including, but are not limited to, powders or granules, suspensions or solutions in water or a non-aqueous medium, 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 may adjust the 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” means delivery of the composition to the nose and nasal cavity through one or both nostrils, and may include delivery by aeration of the composition by a spraying mechanism (device) or a droplet mechanism (device), or by using, for example, a nasal spray, atomizer, dropper, or syringe. Administration of the composition by inhalation may be carried out via the nose or mouth by a spraying or droplet mechanism. As used herein, “inhaler” may be a spraying or droplet device for delivering the composition to the nasal cavity and upper and / or lower respiratory tract of a subject. Delivery may also be carried out directly to any area of ​​the respiratory system (e.g., the lungs) via endotracheal intubation. Those skilled in the art who monitor the clinical response of a subject may adjust the frequency and dosage of drug administration according to methods known in the art.

[0159] In another example, the compositions of the present invention may be administered by topical intranasal administration (intranasal) or by inhalation. As used herein, “topical intranasal administration” means delivery of the composition to the nose and nasal cavity through one or both nostrils, and may include delivery by a spray mechanism (device) or a droplet mechanism (device), or by aerosolization of the composition. Administration of the composition by inhalation may be carried out via the nose or mouth by a spray or droplet mechanism. As used herein, “inhaler” may be a spray or droplet device for delivering the composition in a pharmaceutically acceptable carrier to the nasal cavity and / or upper and / or lower respiratory tract of a subject. Delivery may also be carried out directly to any area of ​​the respiratory system (e.g., the 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 specific composition used, and its mode of administration. Therefore, it is not possible to specify an exact amount for all compositions. However, an appropriate amount can be determined by a person skilled in the art using only routine experiments, taking into account the teachings herein.

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

[0161] In another example, the composition of the present invention, when used, can be administered by parenteral administration of the composition, which generally features injection. The injectable can be prepared in conventional forms as either a liquid solution or suspension, a solid form suitable for dissolution or suspension in a liquid prior to injection, or an emulsion. Modified approaches for parenteral administration include the use of delayed release or sustained release systems so that a constant dosage is maintained.

[0162] In another example, the composition of the present invention can be administered intramuscularly to a subject, for example, by using intramuscular injection or electroporation. Those skilled in monitoring the clinical response of a subject can adjust the frequency and dosage of the drug according to methods known in the art.

[0163] In one example, the composition of the present invention can be administered via a lung lavage procedure. As used herein, "lung lavage" or "total lung lavage" is a procedure where a double-lumen endotracheal tube isolates one lung and sufficient saline or other pharmaceutically acceptable carrier is injected into it to fill the entire volume of one lung. Further, lung lavage can be used to deliver the composition in a pharmaceutically acceptable carrier to the subject's lung / lower airways. Drainage may be performed from the lung being lavaged, filled repeatedly with fluid, and finally aspirated the required number of times. This procedure is repeated on the other lung at another time. In some embodiments, the administration of the composition via lung lavage is extremely severe and may be used for patients admitted to the intensive care unit (ICU).

[0164] In another example, the composition of the present invention may be placed in or stored in a container, bag, pack, or dispenser along with instructions for administration. For example, the instructions may include directions for administering the composition to the target.

[0165] The following patents and applications, their entire specifications incorporated herein by reference: 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 referenced herein are incorporated by reference in their entirety for all purposes. In the event of any conflict, this specification, including the explanation of terms, shall prevail.

[0167] In short, 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, and intermediate compounds thereof. The compounds and compositions are clinically highly effective in crossing the blood-brain barrier (BBB) ​​and acting as tubulin-targeted therapy specific to brain cancer, significantly depolymerizing tubulin to halt cell division, kill cancer cells, and reduce tumor growth. While we do not wish to limit the present invention to any theory or mechanism, the technical features of the present invention are considered to 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 characterized by the administration of the compounds and compositions herein for treating, and / or managing, and / or preemptively preventing, and / or reducing, and / or significantly decreasing cancers, including but not limited to glioblastoma and other brain cancers. In some embodiments, the compounds, compositions and methods relate to the treatment of cancers, including, but not limited to, cancers that metastasize to the brain. In other embodiments, the compounds, compositions and methods relate to the treatment of cancers, including, but not limited to, brain cancer, breast cancer, skin cancer, metastatic cancer, pancreatic cancer, lung cancer, kidney cancer, liver cancer, bladder cancer, osteosarcoma, ovarian cancer, rectal cancer, hematological cancer, gastrointestinal cancer, medulloblastoma, or any combination thereof.

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

[0170] Referring here to Figures 1-54, the present invention features compounds, compositions, and methods for the creation and use of small molecules for the treatment of health conditions, including but not limited to cancer. In some embodiments, 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 cancers, including but not limited to glioblastoma and other brain cancers. In further embodiments, compounds, compositions, and methods relate to drug screening and other methods for the creation of novel compounds.

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

[0172] Embodiments disclosed herein relate to novel chemical compounds for use in treating or preventing health conditions. 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 destabilizers can be used to treat, prevent, or reduce the risk of developing health conditions. In certain embodiments, health conditions may include cancer or other health conditions.

[0173] Some embodiments disclosed herein relate to agents, methods, and processes for preparing the disclosed compounds and compositions containing at least these disclosed compounds. While specific references to individual and collective combinations and permutations of these compounds may not be expressly disclosed, it is understood that the combinations, subsets, interactions, and agents disclosed herein are each intended to be used in a specific manner. Further embodiments of this disclosure are described below.

[0174] I. Compound

[0175] In certain embodiments, the Disclosure provides compounds for use in treating, preventing, or improving a health condition in a subject, or for use as an adjunct therapy in treating, reducing the onset of, or improving a health condition in a subject that requires such treatment or improvement.

[0176] In certain embodiments, the present invention features a compound for use in treating, preventing, or improving a subject's health condition, or for use as a combination therapy in treating, reducing the onset of, or improving a subject's health condition that requires it.

[0177] For example, in some embodiments, the present invention provides a compound having the formula (I-A), ((A m -B n ) j -C-[L’] q ) k -E-L” s -G t (Formula I-A) or an analog, isomer, or pharmaceutically acceptable salt thereof, wherein, A is H, alkyl, alkylcarbonyl, carboxyl, carboxylic acid 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; 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 or bicyclic moiety optionally substituted by one or more heteroatoms or substituents; when n = 0, C is attached to A, L’ is a linker comprising at least one thioester, ester, -CONH-, or -SO2NH-, -NHCO-, or 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 or bicyclic moiety optionally substituted by one or more heteroatoms or substituents; when q = 0, E is attached to C, L'' is the linker, where L'' is O, CH2, -S- or -C=O, s is either 0 or 1, G is [ka] And, t is either 0 or 1, L'' is bonded to a carbon in one of the rings of G, instead of one of R17, R18, R19, R20, R21, R22, or R23. T, U, and V are CH or N, respectively. W is O, S, -CH(R24) or -CH(OR25), and R24 and R25 are 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, linear or branched substituted or unsubstituted alkyl groups, substituted or unsubstituted aryl groups, substituted or unsubstituted heteroaryl groups, and substituted or unsubstituted heterocycles.

[0178] In some preferred embodiments, L'' is bonded to the carbon of G instead of one of R17, R22, or R23.

[0179] In some embodiments, the present invention also uses formula (II-B): [ka] The compound is characterized by having the formula shown, where J is one of the following: [ka] [ka]

[0180] In certain embodiments, the compounds of this disclosure may 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 formula (III-A): [ka] The compound is characterized by the following, where K is one of the following: [ka] [ka]

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

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

[0185] The present invention relates to formula (IV-A): [ka] The compound is characterized by the following, where K is one of the following: [ka] [ka]

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

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

[0188] The present invention also uses formula (VA): [ka] The compound is characterized by the following, where L is one of the following: [ka] [ka]

[0189] In certain embodiments, the compounds of this disclosure may 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 includes formula (VI-A): [ka] The compound is characterized by the following, where L is one of the following: [ka] [ka]

[0192] In certain embodiments, the compounds of this disclosure may 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 formula (VII-A): [ka] The compound is characterized by the following, where L is one of the following: [ka] [ka]

[0195] In certain embodiments, the compounds of this disclosure may 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 present invention is characterized by a compound comprising one of the compounds provided in Table 7, or a derivative thereof.

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

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

[0200] In some embodiments, the compound is of formula IB: ABC-L'-EL”-G (Formula IB) It can include compounds by the formula, in which, A is a linear or branched alkyl, cycloalkyl, or cycloalkylmethyl molecule. B is either O or S, and B is covalently bonded to any available atom on A. C is a cycloalkyl or bicyclic alkyl, and any available atom on C is covalently bonded to B. L' is a thioester, ester, -CONH-, or -SO2NH-, -NHCO-, and any available atom on L' is covalently bonded to any available atom on C. E is a 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] In the formula, any available atom on G is covalently bonded to L'', and L'' can bond to one carbon in the ring of G instead of any one of R17, R18, R19, R20, R21, R22, or R23, and any position not bonded to L'' remains specified. T and U are CH or N, V is CH or N, W is O, S, -CH(R24) or -CH(OR25), and R24 and R25 are 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, linear or branched substituted or unsubstituted alkyl groups, substituted or unsubstituted aryl groups, substituted or unsubstituted heteroaryl groups, and substituted or unsubstituted heterocycles.

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

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

[0203] In some embodiments, the compound may include a compound according to formula II-B or a derivative thereof. [ka] In the formula, 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, and substituted or unsubstituted heteroaryl. Z is selected from the group consisting of -CH or N. R1 is [ka] In the formula, Y is selected from the group consisting of O, S, -C=O, and halides, and if Y is a halide, R13 does not exist. R13 is selected from the group consisting of H, linear or branched substituted or unsubstituted alkyl groups, substituted or unsubstituted aryl groups, substituted or unsubstituted heteroaryl groups, and substituted or unsubstituted heterocycles. BB is a two-ring bridge section. R14 and R15 are each independently selected from the group consisting of H, F, Cl, I, Br, linear or branched substituted or unsubstituted alkyls, substituted or unsubstituted aryls, substituted or unsubstituted heteroaryls, and substituted or unsubstituted heterocycles. R16 is -CO- or -SO2-, R2 is [ka] Therefore, in formula II-B, O is covalently bonded to one carbon in the ring of R2 instead of one of R17, R18, R19, R20, R21, R22, or R23, and any position not bonded to R2 can remain specified. T and U are CH or N, V is CH or N, W is O, S, -CH(R24) or -CH(OR25), and R24 and R25 are 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, linear or branched substituted or unsubstituted alkyl groups, substituted or unsubstituted aryl groups, substituted or unsubstituted heteroaryl groups, and substituted or unsubstituted heterocycles.

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

[0205] [Table 9]

[0206] In some embodiments, the compound may include the compound according to formula III-B or a derivative thereof. [ka] During the ceremony, R5 is [ka] And, R6 [ka] And, In formula III-B, Z is covalently bonded to one of the carbon atoms of the ring R6 instead of one of R17, R18, R19, R20, R21, R22, or R23, and any position not bonded to R6 can remain as specified. 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 groups, substituted or unsubstituted aryl groups, substituted or unsubstituted heteroaryl groups, and substituted or unsubstituted heterocycles. R12 and R13 are each independently selected from the group consisting of H, linear or branched substituted or unsubstituted alkyl groups, substituted or unsubstituted aryl groups, substituted or unsubstituted heteroaryl groups, and substituted or unsubstituted heterocycles. M is selected from the group consisting of O, S, and N (R26), and R26 is selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl. Q is selected from the group consisting of O, S, and halides, and if Z is a halide, then R6 does not exist. Y is selected from the group consisting of O, S, -C=O, and halides. If Y is a halide, then R13 does not exist. BB is a two-ring bridge section, W is selected from the group consisting of O, S, -CH(R24), and -CH(OR25), and 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) may 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 present invention may be any one of the following: [ka]

[0210] In some embodiments, the compound is of formula (IC), [ka] This may include compounds, analogues, isomers, pharmaceutically acceptable salts and / or prodrugs, and / or formulations thereof. In the formula, R1 is [ka] [ka] Selected from, R2 is alkyl, aryl, and heteroaryl, and each aryl and heteroaryl may independently be unsubstituted or independently 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, R1 of the compound of formula (IC) disclosed herein is selected from the following: [ka] [ka]

[0212] In some embodiments, R1 of the compound of formula (IC) disclosed herein is selected from the following: [ka] [ka]

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

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

[0215] In some embodiments, the compounds of the present disclosure include compounds of formula (IC), analogs, isomers, pharmaceutically acceptable salts, and / or prodrugs, where R1 is [ka] [ka] Selected from, R2 is -CH3, [ka] [ka] Selected from, R3 is selected from H and CH3, and R4 is selected from H and F.

[0216] In certain embodiments, the compound of formula (IC) may have R1, R2, R3 and / or R4 as shown in Table 11. In other embodiments, the compounds of the present disclosure may 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, the provided compound is of formula (II-C), [ka] This includes compounds, analogs, isomers, pharmaceutically acceptable salts and / or prodrugs, and / or formulations thereof, wherein R5 is [ka] [ka] Selected from, R6 is alkyl, aryl, and heteroaryl, where each aryl and heteroaryl may independently be unsubstituted or independently 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.

[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 may be compounds of formula (II-C), analogs, isomers, pharmaceutically acceptable salts, and / or prodrugs, and / or formulations thereof, where R5 is [ka] [ka] Selected from, The R6 is -CH3, [ka] [ka] Selected from.

[0225] In certain embodiments, the compound of formula (II-C) may have R5 and / or R6 as shown in Table 12. In other embodiments, the compounds of the present disclosure may 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 is of formula (III-C), [ka] This may include compounds, their analogues, isomers, pharmaceutically acceptable salts and / or prodrugs, In the formula, R7 is [ka] [ka] Selected from, R8 is alkyl, aryl, and heteroaryl, and each aryl and heteroaryl may independently be unsubstituted or independently 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, R 10 It is selected from H or F.

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

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

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

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

[0232] In some embodiments, R8 of the compound 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] That is the case.

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

[0235] In certain embodiments, the compound of formula (III-C) is R7, R8, R9 and / or R shown in Table 13. 10 It can have.

[0236] [Table 13]

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

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

[0239] In some embodiments, the compounds according to this disclosure are 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidine-5-yl)oxy)phenyl)cyclobutan-1-carboxamide, [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compounds disclosed herein have an EC ratio in the range of about 0.2 μM to about 1.0 μM in cancer cell viability. 50 (For example, decreased survival rate, increased tumor cell death), and can have a kinetic solubility in the range of approximately 0.1 μM to approximately 0.5 μM.

[0240] In some embodiments, the compounds according to the present disclosure are 3-methoxy-N-(5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide, [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof.

[0241] In some embodiments, the compound is 3-methoxy-N-(3-methyl-4-((2-morpholinothiazole-5-yl)oxy)phenyl)cyclobutan-1-carboxamide, [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compound has an EC of approximately 0.5 μM to 1.0 μM in cancer cell viability. 50 It has a kinetic solubility in the range of approximately 20.0 μM to approximately 25.0 μM.

[0242] In some embodiments, the compound is 3-methoxy-N-(5-((2-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide, [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compound has an EC of about 0.5 μM to about 2.0 μM in cancer cell viability. 50 It has.

[0243] In some embodiments, the compound is 3-methoxy-N-(4-((2-(4-methoxypiperidine-1-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)cyclobutan-1-carboxamide, [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compound has an EC of about 0.2 μM to about 1.0 μM in cancer cell viability. 50It has a kinetic solubility in the range of approximately 4.0 μM to approximately 7.0 μM.

[0244] In some embodiments, the compound is 3-methoxy-N-(5-(4-morpholinophenoxy)thiazole-2-yl)cyclobutan-1-carboxamide, [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compound has an EC of about 0.1 μM to about 1.0 μM in cancer cell viability. 50 It has a kinetic solubility in the range of approximately 20.0 μM to approximately 30.0 μM.

[0245] In some embodiments, the compound is N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxycyclobutane-1-carboxamide [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compound has an EC of about 0.5 μM to about 1.0 μM in 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)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxycyclobutan-1-carboxamide [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compound has an EC of about 0.5 μM to about 2.0 μM in 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)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compound has an EC of about 0.001 μM to about 0.02 μM in cancer cell viability. 50 It has a kinetic solubility in the range of approximately 85.0 μM to approximately 100.0 μM.

[0248] In some embodiments, the compound is N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compound has an EC of about 45.0 μM to about 55.0 μM in cancer cell viability. 50 It has.

[0249] In some embodiments, the compound is 3-methoxy-N-(5-((2-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide, [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compound has an EC of about 0.05 μM to about 1.0 μM in cancer cell viability. 50 It has a kinetic solubility in the range of approximately 4.0 μM to approximately 6.0 μM.

[0250] In some embodiments, the compound is N-(5-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compound has an EC of about 0.01 μM to about 0.5 μM in cancer cell viability. 50 It has a kinetic solubility in the range of approximately 80.0 μM to approximately 90.0 μM.

[0251] In some embodiments, the compound is N-(4-((2-(2-oxa-7-azaspiro[4.4]nonane-7-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compound has an EC of about 10.0 μM to about 20.0 μM in cancer cell viability. 50 It has.

[0252] In some embodiments, the compound is 3-methoxy-N-(5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide, [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compound has an EC of about 0.01 μM to about 1.0 μM in cancer cell viability. 50 It has a kinetic solubility in the range of approximately 4.0 μM to approximately 25.0 μM.

[0253] In some embodiments, the compound is 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidine-5-yl)oxy)phenyl)bicyclo[1.1.1]pentan-1-carboxamide, [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof.

[0254] In some embodiments, the compound is 3-methoxy-N-(5-(4-morpholinophenoxy)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide, [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compound has an EC of about 0.001 μM to about 0.05 μM in cancer cell viability. 50 It has.

[0255] In some embodiments, the compound is 3-methoxy-N-(5-(4-(piperidine-1-yl)phenoxy)thiazole-2-yl)cyclobutan-1-carboxamide, [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, or any combination thereof. In some embodiments, the compound has an EC of about 1.0 μM to about 2.0 μM in cancer cell viability. 50 It has.

[0256] In some embodiments, the compound is 3-methoxy-N-(5-((6-morpholinopyridine-3-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide, [ka] These include its analogues, isomers, pharmaceutically acceptable salts, prodrugs, polymorphs, 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 Disclosure may include any isomer of any of the compounds disclosed herein and may be designated as having a “cis” or “trans” configuration. According to these embodiments, the compounds of the Disclosure may be any cis or trans isomer of any of the compounds or formulas disclosed herein.

[0259] In some embodiments, the compounds of the present disclosure may contain carbon atoms asymmetrically substituted in R or S configurations, where the terms “R” and “S” are defined as known in the art. According to some embodiments disclosed herein, compounds having asymmetrically substituted carbon atoms having equal amounts of R and S configurations may be racemates at those carbon atoms. In certain embodiments, the present disclosure may include racemic mixtures, relative stereoisomers and absolute stereoisomers, and / or mixtures of relative stereoisomers 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” as herein may refer to a compound produced by modification with a functional group (e.g., a hydroxyl group, a carboxyl group, an amino group, etc.). Examples of “esters” include “esters formed by a hydroxyl group” and “esters formed by a carboxyl group.” The term “ester” may mean an ester whose ester residue is a “conventional protecting group” or a “protecting group that can be removed in vivo by a biological method such as hydrolysis.” In some embodiments, the term “conventional protecting group” may mean a protecting group that can be removed by a chemical method, such as hydrolysis, hydrolysis, electrolysis, or photolysis. In other embodiments, the term “protecting group that can be removed in vivo by a biological method such as hydrolysis” may mean a protecting group that can be removed in vivo by hydrolysis or the like to produce a free acid or a salt thereof after administration to a subject.

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

[0262] In some embodiments, salt derivatives of the compounds disclosed herein formed with a basic group may include, but are not limited to, hydrogen halides, e.g., hydrofluorides, hydrochlorides, hydrobromids, or hydroiodides; inorganic salts, e.g., nitrates, perchlorates, sulfates, or phosphates; lower alkanesulfonates, e.g., methanesulfonates, trifluoromethanesulfonates, or ethanesulfonates; arylsulfonates, e.g., benzenesulfonates, or p-toluenesulfonates; organic salts, e.g., acetates, malates, fumarates, succinates, citrates, ascorbicates, tartrates, oxalates, or maleates; and amino acid salts, e.g., glycine salts, lysine salts, arginine salts, histidine salts, ornithine salts, glutamates, or aspartates. In certain embodiments, pharmaceutically acceptable salts of the compounds disclosed herein may absorb water upon continued exposure to air or recrystallization to form hydrates for use in the formulations disclosed herein.

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

[0264] In certain embodiments, the compounds of this disclosure may exist in crystalline forms including a variety of their solvates and may 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 the packing, geometric arrangement, and / or other descriptive properties of their crystalline solid state. Thus, polymorphs may have different physical properties such as shape, density, hardness, deformability, stability, and solubility. Polymorphs typically exhibit different melting points, IR spectra, and X-ray powder diffraction patterns, NMR signatures, which can be used for identification. In certain embodiments, the compounds of this disclosure may be polymorphic. In certain embodiments, the compounds of this disclosure may 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 herein can be produced by changing and / or adjusting the reaction conditions and / or reagents used in the preparation of the compounds. For example (but not limited to), changes in temperature, pressure, or solvent can result in polymorphism. In some embodiments, different polymorphs of the compounds disclosed herein can be spontaneously converted to other polymorphs.

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

[0266] In other embodiments, the compounds disclosed herein may have a kinetic solubility of at least about 0.35 μM. In some embodiments, the compounds disclosed herein may 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 in between). In yet another embodiment, the compounds disclosed herein can maintain 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 in between or greater than that). In yet another embodiment, the compounds disclosed herein can maintain kinetic solubility for about 1 hour to about 48 hours or longer (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 greater than 48 hours). In some embodiments, the compounds disclosed herein can maintain 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 in between or above) 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 in between or longer).

[0267] In yet another embodiment, 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 in between). In some embodiments, the compounds disclosed herein can maintain kinetic solubility in the range of about 0.39 μM to about 92 μM (e.g., concentrations of 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 in between) at temperatures in the range of about 4°C to about 80°C (e.g., temperatures of 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 in between).

[0268] In some embodiments, the compounds disclosed herein can maintain kinetic solubility in the range of about 0.35 μM to about 92 μM at room temperature (i.e., 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 kinetic solubility in the range of about 0.35 μM to about 92 μM at room temperature (i.e., 25°C ± 5°C) Kinetic solubility in the range of approximately 0.39 μM to approximately 91.23 μM (e.g., approximately 0.39 μM, approximately 1 μM, approximately 5 μM, approximately 10 μM, approximately 20 μM, approximately 30 μM, approximately 40 μM, approximately 50 μM, approximately 60 μM, approximately 70 μM, approximately 80 μM, and approximately 90 μM) can be maintained for approximately 1 hour to approximately 48 hours (e.g., approximately 1 hour, approximately 3 hours, approximately 6 hours, approximately 12 hours, approximately 24 hours, approximately 36 hours, 48 ​​hours, or a timing 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. In some embodiments, the non-human subject is livestock, companion animals, laboratory animals, or zoo animals, wild animals, reptiles, fish, or birds.

[0270] In some embodiments, the compound has an EC50 of 0.005 μM to 50 μM to inhibit the survival rate of cancer cells or induce cancer cell death. In some embodiments, the compound has an EC50 of about 0.0001 μM to 50 μM to inhibit the survival rate of cancer cells or induce cancer cell death. In some embodiments, the compound has an EC50 of about 0.0001 μM to 50 μM to inhibit the survival rate of cancer cells or induce cancer cell death. In other embodiments, the compound has an EC50 of about 0.0001 μM to 15 μM to inhibit the survival rate of cancer cells or induce cancer cell death. In other embodiments, the compound has an EC50 of about 0.0001 μM to 50 μM to inhibit the survival rate of cancer cells or induce cancer cell death.

[0271] In certain embodiments, the compounds disclosed herein may be analyzed and have a Central Nervous System Multi-Parameter Optimization (CNS MPO) score. According to these embodiments, the CNS MPO score is determined using a weighted scoring function based on six basic physicochemical properties: (a) lipophilicity, calculated partition coefficient (ClogP); (b) calculated partition coefficient at approximately pH 7.4 (ClogD); (c) molecular weight (MW); (d) topological polar surface area (TPSA); (e) number of hydrogen bond donors (HBD); and (f) maximum basicity center (pK). a This can be determined by an algorithm that evaluates the following: In these examples, the algorithm assigns a total score ranging from 0 to 6 to the selected compound, where a higher CNS MPO score indicates the compound's ability to cross the blood-brain barrier (BBB). In some embodiments, the compounds disclosed herein may have a CNS MPO score indicating BBB permeability. In other embodiments, the compounds disclosed herein may have a CNS MPO score of 4.0 or higher, indicating their ability to cross the BBB when used to treat a health condition affecting the brain in question.

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

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

[0274] In certain embodiments, as described in a later paragraph, the compounds disclosed herein can reduce the viability of at least one cancer cell or induce cell death. In some embodiments, the compounds disclosed herein can reduce the viability 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 the compounds or mixtures of compounds disclosed herein or their formulations. 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 the compounds or mixtures of compounds disclosed herein or their formulations. In further embodiments, the compounds disclosed herein can reduce tumor metastasis in a subject by about 10% to up to 100% compared to cancer cells not exposed to the compounds or mixtures of compounds or formulations disclosed herein. In some embodiments, the compounds disclosed herein are used to reduce cancer cell viability, decrease tumor volume, and / or reduce tumor metastasis at a half-effective concentration (i.e., EC) of at least about 0.001 μM. 50) may have. In some embodiments, the compounds disclosed herein may have an EC 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 to reduce or inhibit the survival rate of cancer cells, or to kill cancer cells, to reduce tumor volume, and / or to reduce tumor metastasis. 50 It can have.

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

[0276] In other embodiments, the compounds disclosed herein are effective in treating health conditions. In some embodiments, the compounds are effective in preventing the division of cancer cells. In other embodiments, the compounds are effective in inhibiting tubulin polymerization. In other embodiments, the compounds are effective in destabilizing microtubules. In other embodiments, the compounds are effective in arranging G2 / M phase cell division. In some embodiments, the compounds are effective in cytotoxicity against multiple cancer cell lines. In some embodiments, the compounds are effective in treating cancer or metastasis.

[0277] In some embodiments, the compounds disclosed herein are effective in treating health conditions. In some embodiments, the compounds are effective in treating non-neoplastic conditions.

[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 / Conjugate Drugs

[0281] In some embodiments, the present 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 present invention features a prodrug comprising one or more compounds. In other embodiments, the prodrug further comprises 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] Mai. Pharmaceutical composition

[0285] In other embodiments, the Disclosure provides pharmaceutical compositions comprising at least one compound, tagged compound, prodrug, or drug conjugate according to the Disclosure for use in treating a health condition of a subject requiring it. The compounds may be, for example, those described herein.

[0286] In some embodiments, the pharmaceutical composition may include at least one compound disclosed herein and at least one pharmaceutically acceptable carrier. In certain embodiments, the pharmaceutical composition may include a pharmaceutically acceptable carrier, excipient, and / or stabilizer that is nontoxic to the recipient at the dose and / or concentration used to carry out the method disclosed herein.

[0287] In certain embodiments, the weight fraction of the excipient or combination of excipients in the composition may 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 maskers, flavorings, or colorants, as described herein below. The concentrations and types of excipients disclosed herein and used to form the pharmaceutical compositions herein may be selected in accordance with the known principles and knowledge of pharmacy in the art.

[0289] Excipients and other adjuvants

[0290] In some embodiments, the excipient may be a diluent. The diluent may be compressible (i.e., plastically deformable) or abrasively brittle. Not limited 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 acid and butyrate), ethylcellulose, methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, 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 brittle diluents include dibasic calcium phosphate (anhydrous or dihydrate), tribasic calcium phosphate, calcium carbonate, and magnesium carbonate.

[0291] In another embodiment, the excipient may 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, sugars, oligosaccharides, polypeptides, oligopeptides, and combinations thereof.

[0292] In another embodiment, the excipient may be a filler. Suitable fillers include, but are not limited to, carbohydrates, inorganic compounds, and polyvinylpyrrolidone. In non-limiting examples, fillers may 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 may be a buffer. Typical examples of suitable buffers include, but are not limited to, phosphate buffers, carbonate buffers, citrate buffers, Tris buffers, polysaccharide buffers, and buffered salines (e.g., Tris-buffered saline or phosphate-buffered saline). In various embodiments, the excipient may be a pH adjuster. In non-limiting examples, the pH adjuster may be sodium carbonate, sodium bicarbonate, sodium citrate, citric acid, or phosphoric acid. In further embodiments, the excipient may be a disintegrant. The disintegrant may be non-foaming or foaming. Suitable examples of non-foaming disintegrants include, but are not limited to, starches, e.g., corn starch, potato starch, their gelatinized and modified starches, sweeteners, clays, e.g., bentonite, microcrystalline cellulose, alginates, sodium starch glycolate, gums, e.g., agar, guar, carob, karaya, pectin, and tragacanth. Non-limiting examples of suitable foaming disintegrants 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, refined wood cellulose, sodium starch glycolate, isomorphic silicates, and microcrystalline cellulose.

[0295] In another embodiment, the excipient may 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 antimicrobial agents, such as parabens, chlorobutanol, or phenol.

[0296] In further embodiments, the excipient may be a lubricant. Not limited examples of suitable lubricants include minerals, such as talc or silica; and fats, such as vegetable stearin, magnesium stearate, or stearic acid.

[0297] In yet another embodiment, the excipient may be a flavor masking agent. Examples of flavor masking agents include cellulose ether; polyethylene glycol; polyvinyl alcohol; polyvinyl alcohol and polyethylene glycol copolymer; monoglycerides or triglycerides; acrylic polymers; mixtures of acrylic polymers and cellulose ether; cellulose acetate phthalates; and combinations thereof.

[0298] In alternative embodiments, the excipient may be a flavoring agent. The flavoring agent can be selected from synthetic flavoring oils and flavoring agents and / or natural oils, extracts from plants, leaves, flowers, and fruits, and combinations thereof.

[0299] In further embodiments, the excipient may be a colorant. Suitable coloring additives include, but are not limited to, food, drug and cosmetic colorants (FD&C), drug and cosmetic colorants (D&C), or topical and cosmetic colorants (Ext.D&C).

[0300] Other pharmaceutically acceptable carriers, excipients and / or stabilizers are generally non-toxic to the recipient at the dosage and concentration used and are not limited to buffers such as phosphates, citrates, 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; alkylparabens, e.g., methyl or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides Examples include: proteins, e.g., serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, e.g., polyvinylpyrrolidone; amino acids, e.g., glycine, glutamine, asparagine, histidine, arginine, lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextran; chelating agents, e.g., EDTA; sugars, e.g., sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions, e.g., sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants, e.g., TWEEN®, PLURONICS®, or polyethylene glycol (PEG).

[0301] Dosage form

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

[0303] In certain embodiments, the pharmaceutical compositions or formulations disclosed herein may be tablets, pills, capsules, powders, granules, solutions, pastes, ointments or suspensions, patches (e.g., soluble or insoluble), particles, microparticles or nanoparticles or suppositories for unit dosage forms, such as oral, intravenous, subcutaneous, ophthalmic or other drops, or rectal administration, or administration by inhalation or blown-in. In some embodiments, to prepare solid compositions such as tablets or capsules, a main activator (e.g., a compound disclosed herein) can be mixed with a pharmaceutically acceptable carrier, such as a conventional tablet-forming agent, such as 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 the compound according to the present invention or a pharmaceutically acceptable salt thereof. When these pre-formulation compositions are referred to as homogeneous, it means that the active drug or compound is uniformly dispersed throughout the composition, and as a result, the composition can be easily subdivided into effective unit dosage forms such as tablets, pills, and capsules. In some embodiments, the solid pre-formulation compositions disclosed herein are then mixed with the compounds disclosed herein in concentrations 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 It can be subdivided into the above-mentioned unit dosage forms containing 175 mg / kg, approximately 200 mg / kg, approximately 225 mg / kg, approximately 250 mg / kg, approximately 275 mg / kg, approximately 300 mg / kg, approximately 325 mg / kg, approximately 350 mg / kg, approximately 375 mg / kg, approximately 400 mg / kg, approximately 425 mg / kg, approximately 450 mg / kg, approximately 475 mg / kg, approximately 500 mg / kg, or higher, or any concentration in between.

[0304] In some embodiments, the tablets and / or pills disclosed herein can be coated or otherwise formulated to provide a dosage form that offers the advantage of a longer-acting effect. In some embodiments, the tablets and / or pills disclosed herein may have an inner and an outer dosing component, the latter in the form of an envelope covering the former. According to the embodiments herein, the two components may be separated by an enteric coating that serves to resist disintegration in the stomach and allows the inner component to enter the duodenum intact or to be released with delayed release. In certain embodiments, two or more compounds disclosed herein can be mixed together into a single dosage form. In some embodiments, the tablets and / or pills disclosed herein may contain one or more agents that can be used in such an enteric layer or coating, and such agents may include, but are not limited to, several polymer acids, and mixtures of polymer acids with materials such as shellac, cetyl alcohol, and cellulose acetate. Examples of non-limiting surfactants suitable for use herein include nonionic agents or other similar agents, such as polyoxyethylene sorbitan (e.g., Tween® 20, 40, 60, 80, or 85) and other sorbitans (e.g., Span® 20, 40, 60, 80, or 85). In some embodiments, compositions disclosed herein containing a surfactant may have about 0.05% and about 5.0% of the surfactant. In some embodiments, other components, such as mannitol or other pharmaceutically acceptable vehicles deemed appropriate, may be added to the pharmaceutical compositions disclosed herein.

[0305] In some embodiments, the pharmaceutical compositions disclosed herein may be tablets. According to some embodiments herein, the tablets contemplated herein may 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, hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose (HPC), sucrose, gelatin and gum arabic. According to some embodiments herein, the tablets contemplated herein may further contain lubricants such as magnesium stearate, stearic acid, glyceryl behenate and talc. In some embodiments, the pharmaceutical composition may be a solid composition used as a filler in gelatin capsules. According to some embodiments disclosed herein, the excipients contained in the gelatin capsules intended herein may include lactose, starch, cellulose, lactose, or high molecular weight polyethylene glycol, etc.

[0306] In certain embodiments, the pharmaceutical compositions disclosed herein may comprise an emulsion. In some embodiments, the emulsions disclosed herein can be prepared using commercially available lipid emulsions, such as Intralipid®, Liposyn®, Infonutrol®, Lipofandin®, and Lipiphysan®. The active ingredient (e.g., one or more aminopeptidase inhibitors and / or one or more chemotherapeutic agents) may be dissolved in a premixed emulsion composition or in an emulsion formed by mixing oil (e.g., soybean oil, safflower oil, cottonseed oil, sesame oil, corn oil, or almond oil), as well as phospholipids (e.g., egg phospholipid, soybean phospholipid, or soybean lecithin) and water. In some embodiments, other components, such as glycerol or glucose, may be added to the compositions disclosed herein to adjust the tonicity of the emulsion. In other embodiments, the emulsion may contain up to 20% (w / v) of oil, for example, about 5.0% to about 20.0% (w / v). In some embodiments, the emulsion may have lipid droplets of about 0.1 μm to about 1.0 μm and / or have a pH in the range of 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 injection, intracisional injection, intraparenchymal injection, or a combination thereof. In some embodiments, the pharmaceutical compositions disclosed herein formulated for parenteral administration may contain one or more sterile liquids as pharmaceutically acceptable carriers. Non-limiting examples of sterile liquids suitable for use as pharmaceutically acceptable carriers disclosed herein may be water and oils of petroleum, animal, plant, or synthetic origin, including, but not limited to, peanut oil, soybean oil, mineral oil, etc. Saline solutions and aqueous solutions of dextrose and polyethylene glycol (PEG) and glycerol can also be used as liquid carriers, for example, as injection solutions. The pharmaceutical compositions disclosed herein may further contain additional agents, such as preservatives, buffers, isotonic agents, antioxidants and stabilizers, nonionic wetting or clarifying agents, thickeners, etc. In some embodiments, the pharmaceutical compositions disclosed herein may be packaged in single-unit or multiple-dose forms.

[0308] In some embodiments, the 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 include aqueous and non-aqueous sterile suspensions that may include antioxidants, buffers, bacteriostatic agents, and solutes that are isotonic with the blood of the recipient to whom the formulation is intended; as well as, but are not limited to, suspending agents and thickeners. The aqueous solutions may be adequately buffered (e.g., pH of about 3.0 to about 9.0 or about 5.0 to about 8.5 or about 6.5 to about 8.0). Preparation of suitable parenteral formulations under sterile conditions is readily achieved by standard pharmaceutical techniques well known to those skilled in the art.

[0309] In some embodiments, the pharmaceutical compositions described herein may further comprise one or more antimicrobial agents, chemotherapeutic agents, other anticancer therapies, or antibodies or fragments thereof. According to these embodiments, the antimicrobial agent may, for example, be an antiviral, bactericidal, antifungal, or antimicrobial agent, or other antimicrobial agent. For example, the antimicrobial agent may be an antimicrobial agent (antibiotic) such as doxycycline or tetracycline, or other antibiotics, such as generally applicable antibiotics.

[0310] In other embodiments, the Disclosure provides pharmaceutical compositions comprising at least one compound according to the Disclosure for use in treating a health condition of a subject requiring such composition. The compound may be as described, for example, as described herein.

[0311] In some embodiments, the pharmaceutical composition may include at least one compound disclosed herein and at least one pharmaceutically acceptable carrier. In certain embodiments, the pharmaceutical composition may include a pharmaceutically acceptable carrier, excipient, and / or stabilizer that is nontoxic to the recipient at the dose and / or concentration used to carry out the method disclosed herein.

[0312] In certain embodiments, the weight fraction of the excipient or combination of excipients in the composition may 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 maskers, flavorings, or colorants, as described herein below. The concentrations and types of excipients disclosed herein and used to form the pharmaceutical compositions herein may be selected in accordance with the known principles and knowledge of pharmacy in the art.

[0314] IV. Treatment method

[0315] In certain embodiments, the Disclosure provides methods for treating, preventing, reducing the onset of, or improving a health condition in subjects who have, are suspected of developing, or are at risk of developing a health condition. According to these embodiments, the methods may include administering a therapeutically effective amount of the compounds of the Disclosure to a subject, alone or in combination, in the treatment, reduction of onset, or improvement of the health condition in the subject requiring treatment. According to these embodiments, the compounds may be in the form of a therapeutic composition comprising at least one of the compounds of the Disclosure, or a dosage form comprising at least one of the compounds of the Disclosure. In certain embodiments, the health condition may be cancer or non-cancerous growth. In other embodiments, the health condition may 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 described above and herein, respectively.

[0316] cancer

[0317] In some embodiments, the health condition may include, but is not limited to, cancer. Therefore, in certain embodiments, the methods of this disclosure include treating, improving, 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 any other type of 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 carcinoma.

[0318] Certain embodiments disclosed herein relate to treating, improving, preventing, or reducing the risk of developing cancer using at least one of the compounds disclosed herein. Non-limiting examples of cancer may include, but are not limited to, carcinoma, sarcoma, myeloma, leukemia, lymphoma, and mixed types. Other non-limiting examples of cancer include, but are not limited to, connective tissue cancer, bladder cancer, breast cancer, kidney (renal) cancer, lung cancer, lymphoma, pancreatic cancer, prostate cancer, skin cancer, uterine cancer, cancers associated with other organs or tissues, etc. According to these embodiments, the methods of the disclosure may include the use of the compounds disclosed herein to treat, improve, prevent, or reduce the risk of developing any type of cancer.Non-limiting examples of cancer types that are treated, improved, reduced in incidence, or prevented include, but are not limited to, adrenocortical carcinoma, AIDS-related cancers, pathogen-related cancers (e.g., HPV), anal cancer, appendiceal cancer, glioblastoma, medulloblastoma, basal cell carcinoma, bladder cancer, bone cancer, brain tumors, breast cancer, bronchial adenoma / carcinoid, carcinoid tumors (childhood, gastrointestinal), unknown primary carcinomas, cerebellar astrocytoma, cervical cancer, childhood cancer, chronic myeloproliferative disorders, colon cancer, fibrous round cell tumors, endometrial cancer, ependymoma, esophageal cancer, extrahepatic cholangiocarcinoma, cholangiocarcinoma, and ocular cancers (intraocular melanoma, retinal cancer). Blastoma, gallbladder cancer, gastric (stomach) cancer, gastrointestinal stromal tumor, germ cell tumor (extracranial, extragonadal, ovarian), gestational trophoblastic tumor, glioma (adult, pediatric brainstem, pediatric cerebral astrocytoma, pediatric optic tract and hypothalamus, cerebral glioma, malignant glioma), head and neck cancer, hepatocellular (liver) cancer, hypopharyngeal cancer, laryngeal cancer, leukemia (acute lymphoblastic, acute myeloid, chronic lymphocytic, chronic myeloid, hair cell), lip and oral cancer, lung cancer (non-small cell, small cell), lymphoma (AIDS-related, Burkitt, cutaneous T-cell, Hodgkin, non-Hodgkin, primary central nervous system), macroglobulin Waldenström's disease, medulloblastoma (childhood), mesothelioma (adult malignancy, childhood), metastatic squamous cell carcinoma of unknown primary origin, oral cancer, multiple endocrine neoplasm syndrome (childhood), multiple myeloma / plasmacytic neoplasm, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative disorders, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, oropharyngeal cancer, osteosarcoma / malignant fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial carcinoma (surface epithelial stromal tumor), low-grade ovarian tumor, pancreatic cancer, pancreatic cancer (islet cell tumor), parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineal astrocytoma, pineal germ cell tumor, pineoblastoma, This may include supratentorial primitive neuroectodermal tumors (children), pituitary adenomas, pleuroblastomas, primary prostate cancer, rectal cancer, renal cell carcinoma (kidney cancer), rhabdomyosarcoma (children), salivary gland cancers, sarcomas (Ewing family tumors, Kaposi's sarcoma, soft tissue, uterine tumors), Sézary syndrome, skin cancers (non-melanoma, melanoma), cutaneous carcinomas (Merkel cells), squamous cell carcinoma, testicular cancer, pharyngeal cancer, thymoma (children), thymoma and thymic carcinoma, thyroid cancer, thyroid cancer (children), transitional cell carcinoma of the renal pelvis and ureter, primary site of unknown origin (adults, children), urethral cancer, vaginal cancer, vulvar cancer, and Wilms' tumor (children).

[0319] In some embodiments, noncancerous growths can also be treated with at least one compound disclosed herein to reduce progression to cancerous lesions or to suppress the growth of neoplasms or benign tumors, for example. According to these embodiments, the methods of the Disclosure may also include the use of the compounds of the Disclosure to treat, improve, prevent or reduce the risk of developing noncancerous growths in subjects suspected of having or at risk of developing any type of noncancerous growth.

[0320] In certain embodiments, the methods of the present disclosure may involve the use of at least one compound disclosed herein to treat, improve, prevent or reduce the risk of developing cancer at any stage of onset. For example, cancer could be stage 0 cancer, stage I, stage II or stage III (these may be used to describe the number of tumors in the body in question, the size of the tumors, and the extent of metastasis to nearby tissues), or stage IV cancer or metastatic cancer that has spread 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, hematological 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 another embodiment, 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 the paragraphs above, 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, optic tract and hypothalamic glioma. In some embodiments, brain cancer may be glioblastoma, high-grade glioma, other brain cancers, or any combination thereof. In certain embodiments, the methods disclosed herein include the treatment, improvement, or reduction of the incidence of glioblastoma. In other embodiments, the methods disclosed herein include the treatment, improvement, or prevention of the incidence of high-grade glioma.

[0323] In some embodiments, compounds 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 compositions or formulations 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, optic tract and hypothalamic glioma. In some embodiments, the compounds 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 compositions or formulations thereof, can be administered to treat glioblastoma, high-grade glioma, other brain cancers, or any combination thereof. In certain embodiments, the compounds 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 compositions or formulations thereof, can be administered to treat, improve, or reduce the incidence of glioblastoma.In another embodiment, the method disclosed herein includes administering a compound disclosed herein, for example, 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, improve, or prevent the development of a high-grade glioma.

[0324] In other embodiments, RGN6024 or a formulation thereof can be administered to treat brainstem gliomas, cerebellar astrocytomas, cerebral astrocytomas / malignant gliomas, ependymomas, medulloblastomas, supratentorial primitive neuroectodermal tumors, optic tract and hypothalamic gliomas. In some embodiments, RGN6024 or a formulation thereof can be administered to treat glioblastomas, high-grade gliomas, other brain cancers, or any combination thereof. In certain embodiments, RGN6024 or a formulation thereof can be administered to treat, improve, or reduce the incidence of glioblastomas. In other embodiments, the methods disclosed herein include administering the compound RGN6024 or a formulation thereof to treat, improve, or prevent the incidence of high-grade gliomas.

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

[0326] In some embodiments, methods for treating, improving, or preventing a target cancer, metastasis, tumorigenesis or progression, or a combination thereof, may include, but are not limited to, the administration of an effective amount of any of the compounds and / or pharmaceutical compositions disclosed herein. As used herein, “effective amount” means a dose of any of the compounds, formulations and / or pharmaceutical compositions disclosed herein that is sufficient to give a therapeutic effect to a subject having or suspected of having cancer. In certain embodiments, the therapeutic effect on a subject having or suspected of having cancer may include alleviation of the symptoms or consequences of cancer, such as reduction of tumor growth, elimination of tumor cells, prevention or reduction of the development of metastasis from a primary tumor, reduction of the number of tumor cells or tumor volume, reduction or prevention of tumor metastasis, inhibition of the growth or expansion of tumor cells in a primary tumor, secondary tumor and / or metastatic tumor, elimination or elimination of tumor cells, etc.

[0327] Non-neoplastic state

[0328] In some embodiments, a 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 abnormal tissue growth resulting from uncontrolled cell proliferation. Accordingly, in certain embodiments, the methods of this disclosure include the treatment, improvement, or reduction of the risk of developing gout, familial Mediterranean fever, or onychomycosis. In other embodiments, a health condition may include, but is not limited to, a vascular disease.

[0329] subject

[0330] In some embodiments, the subject to be treated for any health condition by any compound or composition disclosed herein may be human. In other embodiments, the subject may be a non-human animal or other mammal. In some embodiments, the subject may be livestock, companion animals, laboratory animals, or zoo animals, wild animals, reptiles, fish, or birds. Non-limiting examples of livestock include, but are not limited to, pigs, cattle, buffalo, goats, sheep, chickens, ducks, geese, turkeys, llamas, and alpacas. Non-limiting examples of companion animals may include pets such as dogs, cats, rabbits, horses, and birds. Zoo animals, as used herein, may include any animals 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 subjects are human subjects, such as fetuses, infants, children, adolescents, young adults, adults, or elderly adults.

[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 that 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 a human. In other embodiments, the subject requiring the methods disclosed herein may be identified by conventional medical examinations, e.g., clinical examinations, biopsies, magnetic resonance imaging (MRI) scans, ultrasound examinations, etc. In some embodiments, the subject treated by the methods described herein may have received, or is receiving, anti-cancer therapies, e.g., chemotherapy, radiotherapy, immunotherapy and / or surgery.

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

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

[0334] Administration

[0335] The compositions of this disclosure can be administered to a subject by any method known in the art. In some embodiments, the compounds 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, buccally, intranasally, by inhalation, intraperitoneally, intrauterine, intratumorally, intravascularly, percutaneously, subcutaneously, rectally, or intrapulmonaryly. Non-limiting examples of administering the compounds 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 compositions or formulations, include intravenous, intramuscular, intrathecal, or intrasternal administration, oral administration, subcutaneous administration, intratumoral, intravascular, intracerebral injection, intracisional, intraventricular, intranasal, or inhalation, parenteral, buccal, enteral, intraperitoneal, inhalable, infusion, intraocular, intravitreous, ocular, rectal, sublingual, topical, percutaneous, intrapulmonary, intrauterine, vaginal, blood-brain barrier disruption by ultrasound, implantable devices, injection techniques, or nanoparticle-based delivery.

[0336] In certain embodiments, the compositions disclosed herein may be administered topically. Topical administration may include the use of transdermal patches or iontophoresis devices. In some embodiments, the compositions disclosed herein may be formulated into dose-unit formulations for administration, further comprising conventional non-toxic, pharmaceutically acceptable adjuvants, carriers, excipients, and vehicles as described herein and above, respectively.

[0337] In certain embodiments, the therapeutic compositions of the Disclosure may be modified to administer concentrations of the compound or mixture of compounds effective in achieving a desired therapeutic response in a particular subject. The selected dose level may depend on a variety of 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 duration of the disease, and the physical condition and medical history of the subject being treated. Determining the appropriate dose for a particular situation is within the scope of the skills of a healthcare professional.

[0338] In some embodiments, the compositions disclosed herein are administered as needed at or immediately before the onset of symptoms. In some embodiments, the compositions are administered regularly according to a prescribed treatment schedule. For example, the compositions of this disclosure can be administered daily 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 another embodiment, the compositions are administered monthly. The compositions may also be administered every 3 to 6 months or at longer intervals if deemed appropriate for long-term administration or treatment. As recognized in the art, the duration of treatment may vary and may be determined by a medical professional.

[0339] The administration of the compositions described herein may also be carried out as part of a treatment regime that may include multiple examples of the administration of one or more compositions or mixtures of compositions. Such regimes may be designed as a method for immediately treating a condition and / or as a method for maintaining the health of a subject in the long term after treatment for a condition (e.g., prevention). For example, a treatment regimen may be designed to delay the onset of symptoms of interest in a subject. It will be understood that the determination of an appropriate treatment regimen is within the scope of the skills of a medical professional.

[0340] Those skilled in the art will also understand that the compositions of this disclosure may be administered co-administered with other therapeutic agents before, after, and / or during treatment with the compositions of this disclosure. The term “co-administered” means administering two or more active ingredients simultaneously with, immediately before, or immediately after the administration of one or more additional therapies. The compounds of this disclosure may be administered alone or co-administered to a subject with another compound or standard agent known in the art. Co-administration means administering compounds individually or in combination simultaneously or sequentially. Dosage and administration intervals can be individually adjusted to provide effective levels of the compound for the specific clinical indication being treated. This provides a treatment regimen that is appropriate to the severity of the individual’s disease condition.

[0341] In certain embodiments, a subject treated with one or more compounds or compositions disclosed herein may have completed, received, or received at least one additional cancer treatment regimen, or an additional cancer treatment regimen before or after surgery administered with the treatment disclosed herein or with the compounds or compositions disclosed herein. In some embodiments, a further treatment regimen of use for treating cancer disclosed herein may include administering one or more anti-cancer therapies or treatments, e.g., chemotherapeutic agents, radiotherapy, small molecules, and immunomodulators. In some embodiments, the anti-cancer therapy or treatment may be administered separately from the compounds or derivatives disclosed herein. In certain embodiments, the anti-cancer therapy or treatment is administered to the subject before, during, or after administration of at least one of the compounds disclosed herein at least once daily, 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 preferred drug regimen.

[0342] In certain embodiments, the present invention further comprises the administration of one or more antimicrobial agents, chemotherapeutic agents, other anticancer therapies, or antibodies or fragments thereof. In some embodiments, the antimicrobial agent comprises one or more antiviral agents, bactericidal agents, antifungal agents, or antibacterial agents, or other antimicrobial agents. In other embodiments, the antibacterial agent comprises one or more doxycycline or tetracycline.

[0343] In some embodiments, the pharmaceutical compositions disclosed herein may be administered before, during, or after at least one chemotherapeutic agent. Non-limiting examples of chemotherapeutic agents may include, but are not limited to, one or more of the following: temozolomide, lomustine, verzutifan, cisplatin, carboplatin, nedaplatin, triplatin tetranitrate, phenantriplatin, 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 or a combination thereof. Non-exclusive examples of HDAC inhibitors include FR01228, trichostatin A, SAHA, and / or PDX101. Non-exclusive examples of DNA synthesis inhibitors include, but are not limited to, capecitabine, floxuridine, decitabine, vidaza, fludarabine, nerarabine, cladribine, clofarabine, pentostatin, tiarabine, troxacitabine, sapacitabine, or forrestin. Further examples of additional chemotherapeutic agents include, but are not limited to, FLT3 inhibitors such as semexanib (SCT5416), sunitinib (SU 11248), midostaurin (PKC412), restatinib (CEP-701), tandutinib (MLN518), CHIR-258, sorafenib (BAY-43-9006), and / or KW-2449. Other non-exclusive examples of additional chemotherapeutic agents include farnesyltransferase inhibitors, such as tipifarnib (Rl 15777, Zarnestra), ronafarnib (SCH66336, Sarasar®), and / or BMS-214662. Other chemotherapeutic agents include, but are not limited to, topoisomerase II inhibitors, such as epipodophyllotoxin (etoposide), teniposide, and the anthracycline doxorubicin and / or 4-epidoxorubicin.Further non-limiting examples of additional chemotherapeutic agents include P-glycoprotein modulators, such as zoskidal trihydrochloride (Z.3HCl), vanadate, or verapamil. Further non-limiting examples of chemotherapeutic agents include hypomethylating agents, such as 5-azacytidine or 2'-deoxyazacitidine.

[0344] In certain embodiments, the pharmaceutical compositions disclosed herein may be administered before, during, or after at least one other drug. 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. Examples of drugs that can be mentioned include, but are not limited to, alectinib, vemurafenib, encorafenib, dabrafenib, olaparib, lucaparib, taazoparib, niraparib, lalotrectinib, entrectinib, lorlatinib, ibrutinib, cobimetinib, binimetinib, trametinib, brigatinib, gilteritinib, ceritinib, ivosidenib, carfilzomib, marizomib, alpelisib, dubellisib, and copanlisib.

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

[0346] In certain embodiments and in earlier paragraphs, at least one additional treatment regimen in the combination therapy disclosed herein may include the administration of radiation. In some embodiments, a subject may be treated by at least one radiotherapy before, during, or after administration of the compounds and / or pharmaceutical compositions disclosed herein. In other embodiments, a subject may be treated by radiotherapy 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 yet another embodiment, a subject may be treated by radiotherapy using ionizing radiation. In some embodiments, a subject may be treated by radiotherapy delivered by a linear accelerator. In further embodiments, a subject may be treated by radiotherapy delivered directly to a tumor. In certain embodiments, radiotherapy can deliver radiation doses in the range of 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 appropriate radiation ranges or doses) directly to the tumor.

[0347] In some embodiments, at least one of the compounds disclosed herein can be administered in conjunction with radiation 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 certain embodiments, the compounds 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 compositions or formulations thereof, can be administered before, during, and / or after radiation to treat a subject having or suspected of having brain cancer.

[0348] In certain embodiments, RGN6024 or a formulation thereof can be administered in conjunction with radiation, either before or after radiation, to treat subjects who have or are suspected of 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 concurrently with or immediately after radiation to treat subjects who have or are suspected of developing brain cancer or other cancers.

[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 having or suspected of having brain cancer (e.g., glioblastoma, high-grade glioma, or cancer that metastasizes to the brain). In certain embodiments, compounds 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 compositions or formulations thereof, can be administered before, during, and / or after at least one chemotherapeutic agent to treat a subject having or suspected of having brain cancer. In some embodiments, at least one of the compounds herein may be administered before, during, and / or after temozolomide to treat subjects who have or are 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 of this specification can be administered before, during, and / or after lomustine to treat subjects who have or are suspected of having brain cancer. In some embodiments, at least one compound of this specification can be administered before, during, and / or after verzutifan to treat subjects who have or are suspected of having brain cancer. In other embodiments, at least one compound of this specification can be administered before, during, and / or after crizotinib to treat subjects who have or are suspected of having non-small cell lung cancer (NSCLC) that has metastasized to the brain. In yet another embodiment, at least one compound of this specification can be administered before, during, and / or after osimertinib to treat subjects who have or are 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 in conjunction with at least one chemotherapeutic agent to treat subjects who have or are suspected of developing 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 subjects who have or are 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 subjects who have or are suspected of having brain cancer. In some embodiments, RGN6024 or a formulation thereof can be administered before, during, and / or after berzutifan to treat subjects who have or are suspected of having brain cancer. In other embodiments, RGN6024 or a formulation thereof can be administered before, during, and / or after crizotinib to treat subjects who have or are suspected of having non-small cell lung cancer (NSCLC) that has metastasized to the brain. In yet another embodiment, RGN6024 can be administered before, during, and / or after osimertinib to treat subjects who have or are suspected of having non-small cell lung cancer (NSCLC) that has metastasized to the brain.

[0352] In some embodiments, at least one compound or formulation thereof disclosed herein can be administered in conjunction with radiation to treat subjects who have or are suspected of developing prostate cancer and / or Ewing's sarcoma. In certain embodiments, the compounds 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 compositions or formulations thereof, can be administered before, during, and / or after radiation to treat subjects who have or are suspected of developing prostate cancer and / or Ewing's sarcoma.

[0353] In certain embodiments, at least one compound disclosed herein may be administered together with at least one inhibitor of ataxia telangiectasia and Rad3-related (ATR) protein to treat subjects who have, have had, are suspected of having, or are at risk of developing prostate cancer and / or Ewing's sarcoma. Non-limiting examples of ATR protein inhibitors (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), etc. In certain embodiments, at least one of the compounds herein can be administered before, during, and / or after Berzosertib to treat subjects who have or are suspected of having prostate cancer and / or Ewing's sarcoma.

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

[0355] In some embodiments, at least one of the compounds disclosed herein can be administered together with at least one PARP1 (poly(ADP)-ribose polymerase-1) inhibitor to treat subjects who have or are suspected of 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, lucaparib, niraparib, talazoparib, and olaparib. In some embodiments, at least one of the compounds disclosed herein can be administered before, during, and / or after olaparib to treat subjects who have or are suspected of developing pancreatic cancer.

[0356] dose

[0357] In some embodiments, the effective dose in humans is approximately 0.5 to 5 mg / kg. In some embodiments, the effective dose in humans is approximately 0.5 to 1 mg / kg. In some embodiments, the effective dose in humans is approximately 1 to 2 mg / kg. In some embodiments, the effective dose in humans is approximately 2 to 3 mg / kg. In some embodiments, the effective dose in humans is approximately 3 to 4 mg / kg. In some embodiments, the effective dose in humans is approximately 4 to 5 mg / kg. In some embodiments, the effective dose in humans is approximately 0.5 to 1.5 mg / kg. In some embodiments, the effective dose in humans is approximately 1 to 1.5 mg / kg. In some embodiments, the effective dose in humans is approximately 2.5 to 3 mg / kg. In some embodiments, the effective dose in humans is approximately 1.5 to 2.5 mg / kg. In some embodiments, the effective dose in humans is approximately 2 to 2.5 mg / kg. In some embodiments, the effective dose in humans is approximately 2.5 to 3.5 mg / kg. In some embodiments, the effective dose in humans is approximately 3 to 3.5 mg / kg. In some embodiments, the effective dose in humans is approximately 3.5 to 4.5 mg / kg. In some embodiments, the effective dose in humans is approximately 3.5 to 5 mg / kg. In some embodiments, the effective dose in humans is approximately 3.5 to 4 mg / kg. In some embodiments, the effective dose in humans is approximately 4 to 4.5 mg / kg. In some embodiments, the effective dose in humans is approximately 4.5 to 5 mg / kg. In some embodiments, the effective dose in humans is approximately 1 to 3 mg / kg. In some embodiments, the effective dose in humans is approximately 3 to 5 mg / kg. In some embodiments, the effective dose in humans is approximately 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 to 25 mg / kg. In some embodiments, the effective dose in mice is 1 to 5 mg / kg. In some embodiments, the effective dose in mice is 5 to 10 mg / kg. In some embodiments, the effective dose in mice is 10 to 15 mg / kg. In some embodiments, the effective dose in mice is 15 to 20 mg / kg. In some embodiments, the effective dose in mice is 20 to 25 mg / kg. In some embodiments, the effective dose in mice is 1 to 3 mg / kg. In some embodiments, the effective dose in mice is 3 to 5 mg / kg. In some embodiments, the effective dose in mice is 5 to 8 mg / kg. In some embodiments, the effective dose in mice is 8 to 10 mg / kg. In some embodiments, the effective dose in mice is 10 to 12 mg / kg. In some embodiments, the effective dose in mice is 12 to 15 mg / kg. In some embodiments, the effective dose in mice is 15 to 17 mg / kg. In some embodiments, the effective dose in mice is 17 to 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 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 has been administered. In some embodiments, the effective dose is administered to the subject before at least one compound has been administered. In some embodiments, the effective dose is administered to the subject while at least one compound is being administered (e.g., simultaneously). 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 a week. 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 a month. 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 several other suitable medication regimens.

[0363] In certain embodiments, the Disclosure provides methods for treating, preventing, reducing the onset of, or improving a health condition in subjects who have, are suspected of developing, or are at risk of developing a health condition. According to these embodiments, the methods may include administering a therapeutically effective amount of the compounds of the Disclosure to a subject, alone or in combination, in the treatment, reduction of onset, or improvement of the health condition in the subject requiring treatment. According to these embodiments, the compounds may be in the form of a therapeutic composition comprising at least one of the compounds of the Disclosure, or a dosage form comprising at least one of the compounds of the Disclosure. In certain embodiments, the health condition may be cancer or non-cancerous growth. In other embodiments, the health condition may 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 described herein and above, respectively.

[0364] How to use

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

[0366] In some embodiments, the present invention relates to a drug screening method for identifying therapeutically effective drug candidates for treating a health condition in a subject requiring such treatment, the method comprising: (a) identifying an in vitro or in vivo model of the health condition; (b) administering the 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 therapeutically effective drug candidates.

[0367] In some embodiments, the present invention relates to a method for detecting target cells having abnormal cell division, the method comprising: (a) identifying a sample containing target cells having abnormal cell division; (b) performing an affinity-based assay using a small molecule conjugated with a tag to contact the target cells having abnormal cell division with the small molecule conjugated with a tag; and (c) determining whether the small molecule conjugated with a tag binds to the target cells 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 immuno-based assay, receptor-based assay, antibody-based assay, nanoparticle-based assay, chemical assay, optical assay, or 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, affinity-based assays are colchicine competitive binding assays.

[0368] A method for detecting a protein expressed by target cells having abnormal cell division, the method comprising: (a) identifying a sample containing the protein expressed by target cells having abnormal cell division; (b) performing an affinity-based assay using a small molecule conjugated with a tag to contact the protein expressed by target cells having abnormal cell division with the small molecule conjugated with a tag; and (c) determining whether the small molecule conjugated with a tag binds to the protein expressed by target cells 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 immuno-based assay, receptor-based assay, antibody-based assay, nanoparticle-based assay, chemical assay, optical assay, or kinetic binding assay. In other embodiments, the affinity-based assay is a 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, affinity-based assays are colchicine competitive binding assays.

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

[0370] In other embodiments, a method for detecting or isolating 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 pull-down assay using a small molecule conjugated with a tag to contact the protein expressed by the target cell having abnormal cell division with the small molecule conjugated with a tag; and (c) selectively isolating the protein expressed by the target cell having abnormal cell division. In other embodiments, the small molecule is RGN6024, or a compound or composition disclosed herein or a derivative thereof.

[0371] kit

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

[0373] In some embodiments, the kits disclosed herein may include at least one container. According to embodiments herein, the container may be any container capable of storing at least one compound disclosed herein or at least one composition containing at least one compound, e.g., a tube, vial, bottle, syringe, e.g., a unit dose, bulk package (e.g., a multi-dose package) or sub-unit dose. Instructions supplied with the kit of the present invention may be instructions described on a label or accompanying document (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 accompanying document indicates that the composition is used for the treatment, delay of onset, and / or mitigation of onset of a health condition or disease intended herein. Instructions for carrying out any of the methods described herein may be provided.

[0374] The kits disclosed herein may include appropriate packaging. Appropriate 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 infusion devices such as minipumps, are also contemplated herein. The kits may have a sterile access port (e.g., the container may be an intravenous solution bag or a vial with a stopper that can be punctured by a subcutaneous needle). The containers may also have a sterile access port (e.g., the container may be an intravenous solution bag or a vial with a stopper that can be punctured by a subcutaneous needle). At least one activator in the composition may be a compound disclosed herein.

[0375] The kit may optionally provide additional components, such as buffers and interpretation information. Typically, the kit includes a container and a label or accompanying documentation that is on or combined with the container. In some embodiments, the present invention provides a product comprising the contents of the above-described kit. [Examples]

[0376] example

[0377] The following examples are included to illustrate specific embodiments. Those skilled in the art will understand that the techniques disclosed in the following examples represent techniques that have been found to work well in the implementation of the claimed methods, compositions and apparatus. However, those skilled in the art will understand that, in light of this disclosure, some of the disclosed embodiments can be modified without departing from the spirit and scope of the embodiments of the invention, and similar or comparable results can still be obtained.

[0378] Example 1

[0379] In one exemplary method, 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidine-5-yl)oxy)phenyl)cyclobutan-1-carboxamide (compound B19) was synthesized. Figure 1 shows the chemical equation 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-morpholinopyrimidine-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 under vacuum. The resulting residue was taken out 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 on magnesium sulfate, filtered, and concentrated under vacuum. The residue was purified by HPLC to obtain 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidine-5-yl)oxy)phenyl)cyclobutan-1-carboxamide (56.4 mg, 0.14 mmol, yield: 40.5%) as an off-white solid. LC-MS and HPLC analysis 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-methoxypiperidine-1-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)cyclobutan-1-carboxamide 2-(4-methoxypiperidine-1-yl)pyrimidine-5-ol (compound B46) was synthesized. Figure 2 shows the chemical equation for the synthesis of compound B46.

[0383] A solution of 4-methoxypiperidine (1.50 g, 13.02 mmol), 2-chloropyrimidine-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 x 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 obtain 2-(4-methoxypiperidine-1-yl)pyrimidine-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-methoxypiperidine-1-yl)-5-(2-methyl-4-nitrophenoxy)pyrimidine. A solution of 2-(4-methoxypiperidine-1-yl)pyrimidine-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 under nitrogen at 100°C for 6 hours. The reaction mixture was concentrated and purified by flash column chromatography (petroleum ether:ethyl acetate = 10:1) to obtain 2-(4-methoxypiperidine-1-yl)-5-(2-methyl-4-nitrophenoxy)pyrimidine (460.0 mg, 1.34 mmol) (yield 56.1%) 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-methoxypiperidine-1-yl)pyrimidine-5-yl)oxy)-3-methylaniline. A suspension of 2-(4-methoxypiperidine-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 at room temperature under a hydrogen atmosphere for 16 hours. Insoluble material was filtered off, and the filtrate was concentrated to obtain 4-((2-(4-methoxypiperidine-1-yl)pyrimidine-5-yl)oxy)-3-methylaniline (220.0 mg, 0.70 mmol) (yield 52.2%) as 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-methoxypiperidine-1-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)cyclobutan-1-carboxamide. A solution of 3-methoxycyclobutan-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-methoxypiperidine-1-yl)pyrimidine-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 and brine, dried over Na2SO4, concentrated, and purified by HPLC to obtain 3-methoxy-N-(4-((2-(4-methoxypiperidine-1-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)cyclobutan-1-carboxamide (54.1 mg, yield 34.2%) as a pale yellow solid. NMR and LC-MS analysis 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-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide (compound B120) was synthesized. Figure 3 shows the chemical equation for the synthesis of compound B120.

[0389] 5-((2-chloropyrimidine-5-yl)oxy)thiazole-2-amine. A mixture of 5-bromothiazole-2-amine (1.14 g, 6.4 mmol), 2-chloropyrimidine-5-ol (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  (3 × 200 mL). The combined organic layer was washed with water and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by column chromatography (petroleum ether:ethyl acetate = 1:2) to obtain the desired compound (600 mg, 2.2 mmol) (yield 35.1%) as a yellow solid. NMR and HPLC 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 271.3 (M + H) + .

[0390] 5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-amine. A mixture of 5-((2-chloropyrimidine-5-yl)oxy)thiazole-2-amine (200 mg, 0.87 mmol), morpholine (229 mg, 2.62 mmol), and Cs2CO3 (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 hours. After completion, the reaction product was cooled to room temperature, diluted with water, and extracted with EA. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by HPLC to obtain 5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-amine (100 mg, 0.36 mmol) (yield 40.9%) 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-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide. 3-Methoxycyclobutan-1-carboxylic acid (46.6 mg, 0.36 mmol), 2-(1H-benzotriazole-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-morpholinopyrimidine-5-yl)oxy)thiazole-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 under vacuum. The obtained residue was removed in dichloromethane (25 mL), and the organic phase was washed with water (10 mL), followed by saturated bicarbonate aqueous solution (3 × 10 mL). The organic layer was collected, dried on magnesium sulfate, filtered, and concentrated under vacuum. The residue was purified by HPLC to obtain 3-methoxy-N-(5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)cyclobutan-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)thiazole-2-yl)cyclobutan-1-carboxamide (compound B121) was synthesized. Figure 4 shows the chemical equation for the synthesis of compound B121.

[0394] 5-((2-chloropyrimidine-5-yl)oxy)thiazole-2-amine. A mixture of 5-bromothiazole-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 layer was washed with water and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by column chromatography (petroleum ether:ethyl acetate = 1:2) to obtain the desired compound (600 mg, 2.2 mmol) (yield 35.1%) as a yellow solid. NMR and HPLC analysis was performed as described in the exemplary methods described herein. LC-MS analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ): m / z 271.3 (M + H) + .

[0395] N-(5-(4-bromophenoxy)thiazole-2-yl)-3-methoxycyclobutan-1-carboxamide. 3-methoxycyclobutan-1-carboxylic acid (286.0 mg, 2.2 mmol), 2-(1H-benzotriazole-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)thiazole-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 under vacuum. The obtained residue was taken out in dichloromethane (25 mL), and the organic phase was washed with water (10 mL), followed by saturated aqueous bicarbonate solution (3 × 10 mL). The organic layer was collected, dried on magnesium sulfate, filtered, and concentrated under vacuum. The residue was purified by column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain N-(5-(4-bromophenoxy)thiazole-2-yl)-3-methoxycyclobutan-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-Morophenoxy)thiazole-2-yl)cyclobutan-1-carboxamide. A mixture of N-(5-(4-bromophenoxy)thiazole-2-yl)-3-methoxycyclobutan-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 hours. After completion, the reaction was cooled to room temperature, diluted with water, and extracted with EA. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by HPLC to obtain 3-methoxy-N-(5-(4-morpholinophenoxy)thiazole-2-yl)cyclobutan-1-carboxamide (30 mg, yield 7%) 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 described 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)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxycyclobutane-1-carboxamide (compound B72) was synthesized. Figure 5 shows the chemical equation for the synthesis of compound B72.

[0399] 2-Chloro-5-(2-methyl-4-nitrophenoxy)pyrimidine. To a solution of 2-chloropyrimidine-5-ol (839.1 mg, 6.41 mmol) in DMA (20.0 mL), 1-fluoro-2-methyl-4-nitrobenzene (1.01 g, 6.41 mmol) and potassium carbonate (6.22 g, 19.21 mmol) were added, and the mixture was stirred overnight at 100°C. 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 obtain 2-chloro-5-(2-methyl-4-nitrophenoxy)pyrimidine (110.5 mg, 0.31 mmol) (yield 6.41%) 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)pyrimidine-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane. 3-oxa-8-azabicyclo[3.2.1]octane (34.1 mg, 0.31 mmol) and cesium carbonate (293.4 mg, 0.91 mmol) were added to a DMA (2.0 mL) solution of 2-chloro-5-(2-methyl-4-nitrophenoxy)pyrimidine (110.5 mg, 0.31 mmol). The mixture was stirred at 150°C for 4 hours. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (40.0 mL x 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 obtain 8-(5-(2-methyl-4-nitrophenoxy)pyrimidine-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane (98.1 mg, 0.28 mmol) (yield 68.29%) 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)pyrimidine-5-yl)oxy)-3-methylaniline. A suspension of 8-(5-(2-methyl-4-nitrophenoxy)pyrimidine-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. Insoluble substances were filtered off, and the filtrate was concentrated to obtain 4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)-3-methylaniline (81.2 mg, 0.26 mmol) (yield 92.85%) 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)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxycyclobutan-1-carboxamide. HBTU (118.2 mg, 0.31 mmol) and DIPEA (100.6 mg, 0.78 mmol) were added to a solution of 3-methoxycyclobutanecarboxylic acid (34.1 mg, 0.26 mmol) in DMF (2.0 mL) at room temperature. Reaction mixture 25 o The mixture was stirred at 10°C for 10 minutes. Then, 4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)-3-methylaniline (81.2 mg, 0.26 mmol) was added and the mixture was stirred overnight at room temperature. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (20 mL × 3). The organic layer was washed with brine (30.0 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by HPLC to obtain N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxycyclobutanecarboxamide (25.9 mg, 0.06 mmol) (yield 23.0%) as a white solid. NMR and LC-MS analysis was 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-morpholinothiazole-5-yl)oxy)phenyl)cyclobutanecarboxamide 5 (compound B71) was synthesized. Figure 6 shows the chemical equation for the synthesis of compound B71.

[0405] 5-(2-methyl-4-nitrophenoxy)thiazole-2-amine. To a solution of 2-methyl-4-nitrophenol (2.05 g, 11.17 mmol) in ACN (20.0 mL), 5-bromothiazole-2-amine (2.01 g, 13.07 mmol) and potassium carbonate (4.91 g, 35.51 mmol) were added, and the mixture was stirred at 80°C for 1 hour. 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 obtain 5-(2-methyl-4-nitrophenoxy)thiazole-2-amine (1.51 g, 6.01 mmol) (yield 53.8%) 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)thiazole-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 overnight at 25°C. 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 obtain 2-bromo-5-(2-methyl-4-nitrophenoxy)thiazole (450.5 mg, 1.43 mmol) (yield 50.8%) 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)thiazole-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), morpholine (1.12 g, 12.82 mmol) and DIPEA (489.5 mg, 3.86 mmol) were added and the mixture was stirred at 100°C for 16 hours. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (40.0 mL x 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 obtain 4-(5-(2-methyl-4-nitrophenoxy)thiazole-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 at room temperature under a hydrogen atmosphere for 4 hours. Insoluble material was filtered off, and the filtrate was concentrated to obtain 3-methyl-4-((2-morpholinothiazol-5-yl)oxy)aniline (240.5 mg, 0.82 mmol) (yield 73.6%) 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-morpholinothiazole-5-yl)oxy)phenyl)cyclobutan-1-carboxamide. HBTU (183.5 mg, 0.43 mmol) and DIPEA (444.3 mg, 1.23 mmol) were added to a solution of 3-methoxycyclobutanecarboxylic acid (54.4 mg, 0.41 mmol) in room temperature DMF (2.0 mL). Reaction mixture 25 o The mixture was stirred at 10°C for 10 minutes. Then, 3-methyl-4-((2-morpholinothiazole-5-yl)oxy)aniline (120.2 mg, 0.41 mmol) was added and the mixture was stirred overnight at room temperature. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (20 mL × 3). The organic layer was washed with brine (30.0 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by HPLC to obtain 3-methoxy-N-(3-methyl-4-((2-morpholinothiazole-5-yl)oxy)phenyl)cyclobutanecarboxamide (31.0 mg, 0.07 mmol, yield: 17.0%) as a white solid. NMR and HP-LC analysis 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-morpholinopyridine-3-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide (compound B126) was synthesized. Figure 7 shows the chemical equation for the synthesis of compound B126.

[0412] 5-(benzyloxy)-2-chloropyridine. A solution of 6-chloropyridine-3-ol (1 g, 7.7 mmol) and Cs2CO3 (7.5 g, 23.1 mmol) in ACN (30 mL) was added to the mixture, 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 Na2SO4, 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)pyridine-2-yl)morpholine. A solution of 5-(benzyloxy)-2-chloropyridine (1.66 g, 7.7 mmol), morpholine (1.4 g, 16.2 mmol), Pd2(dba)3 (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 N2. After quenching the reaction, the mixture was purified by flash column chromatography (petroleum ether:ethyl acetate = 8:1) to obtain 4-(5-(benzyloxy)pyridine-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-Molfolinopyridine-3-ol. To a solution of 4-(5-(benzyloxy)pyridine-2-yl)morpholine (2.0 g, 7.4 mmol) in MeOH (20 mL) at room temperature, Pd / C (200 mg) was added and the mixture was 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 obtain 6-Molfolinopyridine-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 / z181.2 (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 hour. 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) (yield 9%). 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. TFA (2 mL) was added 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 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) (yield 99%) 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-morpholinopyridine-3-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide. A solution of 5-((6-morpholinopyridine-3-yl)oxy)thiazole-2-amine (58 mg, 0.2 mmol), 3-methoxycyclobutan-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 x 3). The organic layer was washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by HPLC to obtain 3-methoxy-N-(5-((6-morpholinopyridine-3-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide (36.5 mg, 0.09 mmol) (46% yield). NMR and HP-LC analysis 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-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide (compound B122) was synthesized. Figure 8 shows the chemical equation 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) were added 2-chloropyrimidin-5-ol (470.5 mg, 3.61 mmol) and cesium carbonate (2.91 g, 8.93 mmol), and the mixture was stirred at 60 °C for 16 h. After quenching the reaction, 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) (yield 35.48%) 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 h. The solution was concentrated to give 5-((2-chloropyrimidin-5-yl)oxy)thiazol-2-amine (365.2 mg, 1.59 mmol) (yield 100%) as a yellow solid. NMR and HPLC analyses were 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 229.7 (M + H) + .

[0422] N-(5-((2-chloropyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxycyclobutan-1-carboxamide. HBTU (714.5 mg, 1.88 mmol) and DIPEA (607.2 mg, 4.71 mmol) were added to a solution of 3-methoxycyclobutanecarboxylic acid (205.3 mg, 1.57 mmol) in room temperature DMF (2.0 mL). Reaction mixture 25 o The mixture was stirred at 10°C for 10 minutes. Then, 5-((2-chloropyrimidine-5-yl)oxy)thiazole-2-amine (365.2 mg, 1.59 mmol) was added and the mixture was stirred overnight at room temperature. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (20.0 mL × 3). The organic layer was washed with brine (30.0 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by HPLC to obtain N-(5-((2-chloropyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxycyclobutanecarboxamide (220.3 mg, 0.64 mmol) (yield 41.2%) 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-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)cyclobutan-1-carboxamide. To a solution of N-(5-((2-chloropyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxycyclobutancarboxamide (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 hours. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (50.0 mL x 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 obtain 3-methoxy-N-(5-((2-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)cyclobutanecarboxamide (45.7 mg, 0.12 mmol) (yield 37.85%) as a yellow solid. NMR and LC-MS analysis 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-(piperidine-1-yl)phenoxy)thiazole-2-yl)cyclobutan-1-carboxamide (compound B125) was synthesized. Figure 9 shows the chemical equation for the synthesis of compound B125.

[0426] tert-butyl(5-(4-bromophenoxy)thiazole-2-yl)carbamate. To a DMA (10.0 mL) solution of 4-bromophenol (1.01 g, 5.83 mmol), tert-butyl(5-bromothiazole-2-yl)carbamate (1.63 g, 5.83 mmol) and potassium carbonate (2.41 g, 17.49 mmol) were added, and the mixture was stirred overnight at 100°C. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (50.0 mL x 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 obtain tert-butyl(5-(4-bromophenoxy)thiazole-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-(piperidine-1-yl)phenoxy)thiazole-2-yl)carbamate. To a solution of tert-butyl(5-(4-bromophenoxy)thiazole-2-yl)carbamate (260.5 mg, 0.70 mmol) in dioxane (5.0 mL), 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) were added, and the mixture was stirred at 100°C for 16 hours. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (40.0 mL x 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 obtain tert-butyl (5-(4-(piperidine-1-yl)phenoxy)thiazole-2-yl)carbamate (121.3 mg, 0.32 mmol) (yield 46.15%) 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-(piperidine-1-yl)phenoxy)thiazole-2-amine. To a solution of tert-butyl(5-((2-chloropyrimidine-5-yl)oxy)thiazole-2-yl)carbamate (121.5 mg, 0.32 mmol) in DCM (3.0 mL), TFA (1.0 mL) was added, and the solution was stirred at 25°C for 4 hours. The solution was concentrated to obtain 5-(4-(piperidine-1-yl)phenoxy)thiazole-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-(piperidine-1-yl)phenoxy)thiazole-2-yl)cyclobutan-1-carboxamide. HBTU (272.8 mg, 0.72 mmol) and DIPEA (234.2 mg, 1.82 mmol) were added to a solution of 3-methoxycyclobutanecarboxylic acid (76.5 mg, 0.59 mmol) in room temperature DMF (2.0 mL). Reaction mixture 25 o The mixture was stirred at 10°C for 10 minutes. Then, 5-(4-(piperidine-1-yl)phenoxy)thiazole-2-amine (165.2 mg, 0.59 mmol) was added and the mixture was stirred overnight at room temperature. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (20.0 mL × 3). The organic layer was washed with brine (30.0 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by HPLC to obtain 3-methoxy-N-(5-(4-(piperidine-1-yl)phenoxy)thiazole-2-yl)cyclobutanecarboxamide (28.1 mg, 0.33 mmol) (yield 56.0%) as a white solid. NMR and LC-MS analysis 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)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxycyclobutane-1-carboxamide (compound B130) was synthesized. Figure 10 shows the chemical equation for the synthesis of compound B130.

[0432] tert-butyl(5-((2-chloropyrimidine-5-yl)oxy)thiazole-2-yl)carbamate. A solution of tert-butyl(5-bromothiazole-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-chloropyrimidine-5-ol (600 mg, 4.6 mmol), and the mixture was stirred at 70°C for 4 hours. 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 crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain tert-butyl(5-((2-chloropyrimidine-5-yl)oxy)thiazole-2-yl)carbamate (600 mg, 1.8 mmol) (yield 50%) 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)pyrimidine-5-yl)oxy)thiazole-2-yl)carbamate. A solution of tert-butyl(5-((2-chloropyrimidine-5-yl)oxy)thiazole-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 hours. The reaction mixture was extracted with ethyl acetate (50 mL x 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 obtain tert-butyl(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)carbamate (140 mg, 0.34 mmol) (yield 57%). 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)pyrimidine-5-yl)oxy)thiazole-2-amine. To a solution of tert-butyl(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)carbamate (140 mg, 0.34 mmol) at room temperature in DCM (10 mL), TFA (2 mL) was added and the mixture was stirred at room temperature for 4 hours. The mixture was concentrated to obtain 5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)thiazole-2-amine (107 mg, 0.32 mmol) (yield 99%). 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)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxycyclobutan-1-carboxamide. A solution of 5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)thiazole-2-amine (107 mg, 0.32 mmol), 3-methoxycyclobutan-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 x 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 obtain N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxycyclobutan-1-carboxamide (19.3 mg, 0.04 mmol) (yield 14%). NMR and HPLC analysis was performed as described in the exemplary methods disclosed herein. LC-MS (ESI + ):m / z418.2 (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-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide (compound B140) was synthesized. Figure 11 shows the chemical equation for the synthesis of compound B140.

[0438] tert-butyl(5-((2-chloropyrimidine-5-yl)oxy)thiazole-2-yl)carbamate. To a solution of tert-butyl(5-bromothiazole-2-yl)carbamate (400 mg, 1.44 mmol) in DMF (10 mL), 2-chloropyrimidine-5-ol (222.9 mg, 1.73 mmol) and Cs2CO3 (1.4 g, 4.32 mmol) were added. The mixture was prepared at 100°C. o The mixture was stirred at 12 hours in C. 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 obtain tert-butyl(5-((2-chloropyrimidine-5-yl)oxy)thiazole-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-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)carbamate. A solution of tert-butyl(5-((2-chloropyrimidine-5-yl)oxy)thiazole-2-yl)carbamate (250 mg, 0.76 mmol) in DMF (5 mL) was mixed with piperidine (331.2 mg, 3.8 mmol) and DIPEA (294.1 mg, 2.28 mmol). The mixture was prepared at 100°C. oThe mixture was stirred at 12 hours in 1C. 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 obtain tert-butyl(5-((2-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-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-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-amine. A solution of tert-butyl(5-((2-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)carbamate (180 mg, 0.47 mmol) and TFA (3 mL) in DCM (5 mL) was stirred at 0°C for 3 hours. After quenching the reaction, the solution was concentrated to obtain 5-((2-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-amine (120.0 mg, 0.43 mmol) (yield 99.01%) as a yellow solid together with the TFA salt. 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-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide. To a solution of 5-((2-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-amine (120 mg, 0.43 mmol) in DMF (3 mL), 3-methoxybicyclo[1.1.1]pentan-1-carboxylic acid (50 mg, 0.35 mmol), HBTU (159.9 mg, 0.42 mmol), and DIPEA (136.2 mg, 1.06 mmol) were added. The mixture was stirred at room temperature 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 Na2SO4, filtered, and concentrated. The residue was purified by HPLC to obtain 3-methoxy-N-(5-((2-(piperidine-1-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide (63.9 mg, 0.16 mmol) (yield 45.3%) as a yellow solid. NMR and LC-MS analysis 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-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide (RGN6024) was synthesized. Figure 12A shows the chemical equation for the synthesis of RGN6024.

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

[0445] 5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-amine. 2-morpholinopyrimidine-5-ol (80.0 g, 442 mmol) and 5-bromothiazole-2-amine hydrobromide (229 g, 883 mmol) were dissolved in N,N-dimethylformamide (500 mL), to which Cs2CO3 (431 g, 1.33 mol) was added. 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 x 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under vacuum. The residue was purified by column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain 5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-amine (50.0 g) as a yellow solid. LC-MS (ESI + ):m / z 280.1 [M + H] + .

[0446] 3-Methoxy-N-(5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)bicyclo[1.1.1]pentane-1-carboxamide. HATU (136.1 g, 0.36 mol) was added to a solution of 5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-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). The resulting mixture was stirred under a nitrogen atmosphere at room temperature for 16 hours. The mixture was then poured into water (1.5 L) and extracted with ethyl acetate (250 mL x 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under vacuum. The residue was purified by column chromatography (petroleum ether:ethyl acetate = 1:2) to obtain 3-methoxy-N-(5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide (24.1 g) as a yellowish-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)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide (compound B138) was synthesized. Figure 13 shows the chemical equation for the synthesis of compound B138.

[0449] tert-butyl(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)carbamate. A solution of tert-butyl(5-((2-chloropyrimidine-5-yl)oxy)thiazole-2-yl)carbamate (101.7 mg, 0.31 mmol) in DMF (5 mL) was mixed with piperidine (36.9 mg, 0.31 mmol) and DIPEA (119.9 mg, 0.93 mmol). The mixture was then mixed to a concentration of 100. o The mixture was stirred in 1C 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 obtain tert-butyl(5-((2-morpholinopyrimidine-5-yl)oxy)thiazole-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)pyrimidine-5-yl)oxy)thiazole-2-amine. A solution of tert-butyl(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)carbamate (90 mg, 0.22 mmol) and TFA (3 mL) in DCM (5 mL) was stirred at 0°C for 3 hours. After quenching the reaction, the solution was concentrated to obtain 5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)thiazole-2-amine (65.1 mg, 0.21 mmol) (yield 97.0%) 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)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide. To a solution of 5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)thiazole-2-amine (65.1 mg, 0.21 mmol) in DMF (3 mL), 3-methoxybicyclo[1.1.1]pentan-1-carboxylic acid (40 mg, 0.28 mmol), HBTU (128.3 mg, 0.34 mmol), and DIPEA (109.1 mg, 0.85 mmol) were added. The mixture was stirred at room temperature 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 Na2SO4, filtered, and concentrated. The residue was purified by HPLC to obtain N-(5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide (33.8 mg, 0.08 mmol) (yield 28.0%) as a yellow solid. NMR and LC-MS analysis was 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]nonane-7-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide (compound B137) was synthesized. Figure 14 shows the chemical equation for the synthesis of compound B137.

[0454] tert-butyl(5-((2-(2-oxa-7-azaspiro[4.4]nonane-7-yl)pyrimidine-5-yl)oxy)thiazole-2-y)carbamate. A solution of tert-butyl(5-((2-chloropyrimidine-5-yl)oxy)thiazole-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 under nitrogen 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) and dried over Na2SO4 to obtain tert-butyl(5-((2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)carbamate (180.0 mg, 0.43 mmol) (yield 63.3%) 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]nonane-7-yl)pyrimidine-5-yl)oxy)thiazole-2-amine. A solution of tert-butyl(5-((2-(2-oxa-7-azaspiro[4.4]nonane-7-yl)pyrimidine-5-yl)oxy)thiazole-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 quenching the reaction, the solution was concentrated to obtain 5-((2-(2-oxa-7-azaspiro[4.4]nonane-7-yl)pyrimidine-5-yl)oxy)thiazole-2-amine (132.2 mg, 0.41 mmol) (yield 96.4%) 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]nonane-7-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide. To a solution of 5-((2-(2-oxa-7-azaspiro[4.4]nonane-7-yl)pyrimidine-5-yl)oxy)thiazole-2-amine (132.2 mg, 0.41 mmol) in DMF (3 mL), 3-methoxybicyclo[1.1.1]pentan-1-carboxylic acid (58.2 mg, 0.41 mmol), HBTU (233.1 mg, 0.62 mmol), and DIPEA (158.7 mg, 1.23 mmol) were added. The mixture was stirred at room temperature 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 Na2SO4, filtered, and concentrated. The residue was purified by HPLC to obtain N-(5-((2-(2-oxa-7-azaspiro[4.4]nonane-7-yl)pyrimidine-5-yl)oxy)thiazole-2-yl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide (33.0 mg, 0.08 mmol) (yield 18.6%) as a yellow solid. NMR and LC-MS analysis was 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-morpholinopyrimidine-5-yl)oxy)phenyl)bicyclo[1.1.1]pentane-1-carboxamide (compound B89) was synthesized. Figure 15 shows the chemical equation for the synthesis of compound B89.

[0459] 2-Molfolinopyrimidine-5-ol. A solution of 2-chloropyrimidine-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 Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain 2-morpholinopyrimidine-5-ol (650.0 mg, 3.59 mmol) (yield 58.6%) 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)pyrimidine-2-yl)morpholine. To a solution of 2-morpholinopyrimidine-5-ol (300.0 mg, 1.66 mmol) in DMF (4 mL), 1-fluoro-2-methyl-4-nitrobenzene (257.5 mg, 1.66 mmol) and Cs2CO3 (1.6 g, 4.98 mmol) were added at room temperature. The reaction mixture was stirred at room temperature 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 Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain 4-(5-(2-methyl-4-nitrophenoxy)pyrimidine-2-yl)morpholine (200.0 mg, 0.63 mmol) (yield 51.46%) 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-morpholinopyrimidine-5-yl)oxy)aniline. Pd / C (20 mg) was added to a solution of 4-(5-(2-methyl-4-nitrophenoxy)pyrimidine-2-yl)morpholine (200.0 mg, 0.63 mmol) at room temperature in MeOH (4 mL). 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 obtain 3-methyl-4-((2-morpholinopyrimidine-5-yl)oxy)aniline (120.0 mg, 0.42 mmol) (yield 66.7%) 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-morpholinopyrimidine-5-yl)oxy)phenyl)bicyclo[1.1.1]pentane-1-carboxamide. To a room temperature solution of 3-methyl-4-((2-morpholinopyrimidine-5-yl)oxy)aniline (120.0 mg, 0.42 mmol) in DMF (3 mL), 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) were added. 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 x 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by HPLC to obtain 3-methoxy-N-(3-methyl-4-((2-morpholinopyrimidine-5-yl)oxy)phenyl)bicyclo[1.1.1]pentan-1-carboxamide (78.5 mg, 0.19 mmol) (yield 45.53%) as a yellow oil. NMR and LC-MS analysis 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)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide (compound B155) was synthesized. Figure 16 shows the chemical equation for the synthesis of compound B155.

[0465] 4-Morpholinophenol. A 0°C 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 hours under N2 protection. 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 crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain 4-morpholinophenol (535.1 mg, 2.99 mmol) (yield 51.66%) 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)thiazole-2-yl)carbamate. To a room temperature solution of 4-morpholinophenol (260.0 mg, 1.45 mmol) in DMF (5 mL), tert-butyl(5-bromothiazole-2-yl)carbamate (443.3 mg, 1.59 mmol) and Cs2CO3 (1.4 g, 4.35 mmol) were added. The mixture was heated at 80°C for 4 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 crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain tert-butyl (5-(4-morpholinophenoxy)thiazole-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)thiazole-2-amine. To a solution of tert-butyl(5-(4-morpholinophenoxy)thiazole-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 obtain 5-(4-morpholinophenoxy)thiazole-2-amine (50.0 mg, 0.18 mmol) (yield 81.9%) 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)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide. To a solution of 5-(4-morpholinophenoxy)thiazole-2-amine (50.0 mg, 0.18 mmol) at room temperature in DMF (3 mL), 3-methoxybicyclo[1.1.1]pentan-1-carboxylic acid (25.6 mg, 0.18 mmol), HATU (102.6 mg, 0.27 mmol), and DIPEA (69.7 mg, 0.54 mmol) were added. 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 x 3). The organic layer was washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by HPLC to obtain 3-methoxy-N-(5-(4-morpholinophenoxy)thiazole-2-yl)bicyclo[1.1.1]pentan-1-carboxamide (7.8 mg, 0.02 mmol) (yield 10.8%) as a yellow oil. NMR and LC-MS analysis 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)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide (compound B118) was synthesized. Figure 17 shows the chemical equation 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-chloropyrimidine-5-ol (5.05 g, 38.71 mmol), and potassium carbonate (13.36 g, 96.78 mmol) in DMA (70 mL) was stirred under nitrogen at 100°C 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 Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain 2-chloro-5-(2-methyl-4-nitrophenoxy)pyrimidine (220.0 mg, 0.83 mmol) (yield 2.5%) 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)pyrimidine-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 hours) and then cooled to room temperature. The reaction mixture was purified by flash column chromatography to obtain 8-(5-(2-methyl-4-nitrophenoxy)pyrimidine-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane (210.0 mg, 0.61 mmol) (yield 81.3%) as a yellow solid. NMR and HPLC 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 343.1 (M + H) +

[0473] 4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)-3-methylaniline. A suspension of 8-(5-(2-methyl-4-nitrophenoxy)pyrimidine-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. Insoluble material was filtered off, and the filtrate was concentrated to obtain 4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)-3-methylaniline (150.0 mg, 0.48 mmol) (yield 78%) 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)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide. A solution of 3-methoxybicyclo[1.1.1]pentan-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. Next, 4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-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 obtain N-(4-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide (152.9 mg, 0.35 mmol) (73% yield) as a yellow solid. NMR and LC-MS analysis was 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]nonane-7-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentane-1-carboxamide (compound B117) was synthesized. Figure 18 shows the chemical equation for the synthesis of compound B117.

[0477] 2-(2-oxa-7-azaspiro[4.4]nonane-7-yl)pyrimidine-5-ol. A solution of 2-chloropyrimidine-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 under nitrogen at 80°C for 16 hours. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (50 mL x 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 (DCM:MeOH=93:7) to obtain 2-(2-oxa-7-azaspiro[4.4]nonan-7-yl)pyrimidine-5-ol (150 mg, 0.68 mmol) (yield 29.3%) as 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)pyrimidine-2-yl)-2-oxa-7-azaspiro[4.4]nonane. A solution of 2-(2-oxa-7-azaspiro[4.4]nonane-7-yl)pyrimidine-5-ol (150 mg, 0.68 mmol), 1-fluoro-2-methyl-4-nitrobenzene (105.0 mg, 0.68 mmol), and Cs2CO3 (665.0 mg, 3.04 mmol) in DMF (3 mL) was stirred at room temperature for 3 hours. After quenching the reaction, the reaction mixture was extracted with ethyl acetate (50 mL x 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 = 1:2) to obtain 7-(5-(2-methyl-4-nitrophenoxy)pyrimidine-2-yl)-2-oxa-7-azaspiro[4.4]nonane (150 mg, 0.42 mmol) (yield 61.9%) 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]nonane-7-yl)pyrimidine-5-yl)oxy)-3-methylaniline. A suspension of 7-(5-(2-methyl-4-nitrophenoxy)pyrimidine-2-yl)-2-oxa-7-azaspiro[4.4]nonane (150 mg, 0.42 mmol) in methanol (10 mL) was stirred at room temperature under a hydrogen atmosphere for 16 hours. Insoluble material was filtered off, and the filtrate was concentrated to obtain 4-((2-(2-oxa-7-azaspiro[4.4]nonane-7-yl)pyrimidine-5-yl)oxy)-3-methylaniline (120.0 mg, 0.37 mmol) (yield 81.8%) as 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)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide. A solution of 3-methoxybicyclo[1.1.1]pentan-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. Next, 4-((2-(2-oxa-7-azaspiro[4.4]nonane-7-yl)pyrimidine-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 obtain N-(4-((2-(2-oxa-7-azaspiro[4.4]nonane-7-yl)pyrimidine-5-yl)oxy)-3-methylphenyl)-3-methoxybicyclo[1.1.1]pentan-1-carboxamide (145.5 mg, 0.32 mmol) (yield 87.3%) as a yellow solid. NMR and LC-MS analysis 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, in various cancer cell lines (e.g., human cancer cell lines), several compounds and their isomers disclosed herein are measured at half effective concentrations (EC2). 50 The following was determined. Briefly, cells derived from one prostate cancer cell line (22RV1) and four glioblastoma cell lines (U87, LN-18, LN-229, and T98G) were plated into multiwell plates. After 24 hours, one of the compounds disclosed herein was added to the wells at gradually increasing concentrations. After 72 hours of treatment, cell viability was evaluated using alamarBlue® (Invitrogen), and fluorescence was read using a CLARIOstar Plus Multilabel Plate Reader (BMG Labtech). Samples were normalized to an untreated control. EC 50 The values ​​were determined using GraphPad Prism. EC of the compounds tested in each cell line. 50 The results are provided in Tables 15 and 16. In the tables, "Inactive" means less than 50% inhibition at the highest concentration tested at 50 μM, and "NT" means untested.

[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 multi-parameter optimization (CNS MPO) score, kinetic solubility, and cellular permeability of selected compounds were determined. The CNS MPO scores of the compounds and their isomers disclosed herein were calculated using an algorithm that employs a weighted scoring function that evaluates six key physicochemical properties for blood-brain barrier (BBB) ​​permeability (clogP, clogD, MW, TPSA, HBD, and pKa). The CNS MPO scores range from 0 to 6.0, and a score ≥ 4.0 was used as a cutoff to select compounds that are likely to accumulate in the CNS. The kinetic solubility of a compound is the maximum solubility of the species of compound that precipitates most rapidly. Kinetic solubility was determined by preparing concentrated stock solutions of the compounds and their isomers disclosed herein in an organic solvent (DMSO), then mixing the solutions with PBS buffered aqueous solution, and subsequently filtering. The filtrate was tested to quantify the kinetic solubility using an HPLC-MS calibration curve. Cellular permeability was determined by the apparent permeability coefficient (P app ) and efflux ratio (P app BA / P app AB This was determined by calculating the apparent transmittance coefficient (Papp) using the following formula:

number

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

[0488] [Table 17]

[0489] Example 21

[0490] In another exemplary method, the effects of the compounds of this disclosure on tubulin polymerization were determined. It should be noted 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 effects of compounds B130, B144, and B147 at a 5 μM concentration on tubulin polymerization (Figure 19A) and the effects of compounds B137, B138, and RGN6024 (Figure 19B). Colchicine and nocodazole were positive controls, and DMSO was 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 EBI binding inhibitor using the MCF7 cell line, demonstrating the binding of the compound to the colchicine binding site (Figure 22). Colchicine (COL) was used as a positive control and vinblastine 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, the ICR1 male mouse strain was 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 physiological saline. A single dose of 30 mg / kg of compound B138 or RGN6024 was administered using this method. The concentrations of compound B138 or RGN6024 in plasma were measured using HPLC-MS quantification (Figures 23 and 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, levels of RGN6024 in brain tissue were measured after oral administration. 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 physiological saline. The concentration of RGN6024 in brain tissue administered with a single dose of 30 mg / kg using this method was measured by HPLC-MS quantification (Figure 25). Levels of RGN6024 in plasma and brain were higher than the concentrations required to kill cancer cells in vitro.

[0496] Example 24

[0497] In another exemplary method, RGN6024 was tested in an LN-18 xenograft model of CB17 SCID female mice. For this study, LN-18 cancer cells were transplanted into CB17 SCID mice. Tumor volume was measured on days 2, 4, 6, 8, 10, 13, and 15. RGN6024 was administered orally at a dose of 15 mg / kg (from day 1 to 5) and 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 amounts of the compound RGN6024 in plasma, brain, and tumor were higher than the levels required to kill LN-18 cancer cells.

[0498] Example 25

[0499] In another exemplary manner, RGN6024 is further characterized as shown in the illustration and as described below herein.

[0500] As disclosed herein, it has been discovered that the compounds of this application can be used to treat health conditions such as cancer. Furthermore, it has been observed that many of the compounds can target cell cycle elements and induce cancer cell death. For example, RGN6024 was 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 the compound for 24 hours and then fixed with 4% paraformaldehyde for 20 minutes. After washing the cells with PBS, they were permeabilized with FoxP3 perm buffer (BD Biosciences) at room temperature for 10 minutes. 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 magnification) were obtained using automated high-content imaging microscopes (Operetta, Perkin Elmer).

[0501] Another experiment showed that RGN6024 had an improved ability to cross the blood-brain barrier, with many failing to enter the brain and others having a much lower success rate of crossing the BBB compared to other known products on the market than those observed herein for RGN6024 and its related compounds.

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

[0503] Current FDA-approved therapies related to tubulin interactions have been observed to bind to tubulin at large cleavage points on the protein's surface. Therefore, these drugs are themselves large. Regarding brain penetration, the larger the molecule, the less likely it is to penetrate the brain. Thus, large molecules struggle to cross the blood-brain barrier. If 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 problems with blood-brain barrier permeability but are often pumped back. This increases the dose required to overcome these problems, reducing efficacy and efficiency and increasing the likelihood of toxicity problems.

[0504] As disclosed herein, using different approaches, some molecules of this application have been designed to bind to small cracks on the surface of tubulin protein. This modification allows for smaller molecules, and these properties act to improve brain penetration and evade the efflux pump 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 for brain cancer, LN-18 xenograft tumors and brains were excised from a mouse brain cancer model after the last dose. This was done at two time points: half 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 exceeded the amount required to shrink the tumors in the animals, and therefore, treating these tumors with lower doses is more effective.

[0506] In other exemplary methods, RGN6024 was tested in various high-grade glioma strains, as well as in breast, lung, and melanoma tumors with a tendency to metastasize to the brain. A median potency of 91 nM was observed. In comparison, the brain penetration level of RGN6024 is 100 times higher than the concentrations required to see an effective response in these cancer models.

[0507] Example 26

[0508] In another exemplary method, patients with treatment-resistant glioblastoma may or may be treated with once-daily oral administration of RGN6024, either as a monotherapy or in combination with other treatments. Brain and tumor biopsies or other minimally invasive analyses, such as blood analysis or other fluid analyses, confirm that RGN6024 directly achieves high concentrations in the tumor and brain. Notably, patients will also experience significant tumor reduction. Surprisingly, patients will continue to survive significantly beyond 5 years (the survival rate for patients during this period is only 5% when using currently available treatments).

[0509] Example 27

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

[0511] In some embodiments, the present 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, incorporate RGN6024, or be a combination thereof. In some embodiments, the system may include a processor configured to execute computer-readable instructions and a memory component communicably coupled to the processor. The memory component may include an AI model, which includes one or more neural networks trained on a training dataset containing chemical training data. Training the AI ​​model may include supplying 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 to the AI ​​model. The chemical data may include data specific to RGN6024. The computer-readable instructions may further include the AI ​​model generating one or more candidate compounds.

[0512] In some embodiments, the present 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, incorporate RGN6024, or be a combination thereof. The method may include providing an AI model comprising one or more neural networks trained on a training dataset comprising chemical training data. Training the AI ​​model may include supplying 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, which includes data specific to RGN6024, into the AI ​​model, and having the AI ​​model generate one or more candidate compounds.

[0513] In some embodiments, one or more candidate compounds may comprise one or more candidate drug compounds. In some embodiments, data specific to RGN6024 may comprise 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 present 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, incorporate RGN6024, or be a combination thereof. The AI ​​model may include one or more neural networks. The training method may include supplying a training dataset containing chemical training data specific to RGN6024 as input to the AI ​​model. The method may further include inputting chemical data into the AI ​​model, the chemical data containing data specific to RGN6024. The AI ​​model may be trained to generate one or more candidate compounds as outputs.

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

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

[0517] In some embodiments, chemical training data may include one or more training labels associated with one or more chemical compounds. In some embodiments, one or more training labels may include one or more label elements having predetermined values. In some embodiments, 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 may include data relating to one or more chemical compounds. The data relating to one or more chemical compounds may include one or more structural properties of one or more chemical compounds, one or more physical properties of one or more chemical compounds, one or more interactions between one or more chemical compounds, one or more molecular pathways of one or more chemical compounds, one or more molecular and cellular profiles of 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, chemical data may include one or more labels associated with one or more chemical compounds. In some embodiments, one or more labels may include one or more label elements having predetermined values. In some embodiments, one or more labels may include bioassay results, toxicity, cross-reactivity, pharmacokinetics, pharmacodynamics, bioavailability, solubility data, or a combination thereof.

[0521] AI models can be stored, trained, and / or run 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. AI models may also be stored in the form of program code, as described above. One or more neural networks in an AI model may, in some embodiments, include perceptron neural networks, feedforward neural networks, multilayer perceptron neural networks, radial basis function neural networks, recurrent neural networks, long- and short-term memory neural networks, sequence-to-sequence neural network models, modular neural networks, graph-based convolutional neural networks, instance-based models, feature attribute models, and the like.

[0522] In an unrestricted example, the AI ​​model of the claimed invention may include a perceptron neural network that takes chemical data as input, performs one or more functions on the input, multiplies the outputs of the one or more functions by multiple weights, and produces a final output containing one or more candidate compounds. In another unrestricted example, the AI ​​model of the claimed invention may include a multilayer perceptron neural network comprising multiple layers, each layer 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 to a 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.

[0523] In another non-limiting example, the AI ​​model may include a graph-based convolutional neural network with 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 to a 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 the functions within them may be configured to extract one or more subgraphs from an input graph containing one or more relevant node features, and to apply one or more attention-based functions to the subgraphs and relevant node features to assign importance to specific molecules, bonds, atoms, etc., so that the model can generate and / or predict new candidate compounds based on the input, including RGN6024, by utilizing the most relevant patterns determined by one or more attention-based functions. Determining which patterns are relevant and important for discovery may be components of the training dataset.

[0524] 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 reactions, one or more compounds associated with RGN6024, or a combination thereof.

[0525] With regard to methods for implementing AI for discovering compounds related to RGN6024 and AI-based systems thereto, 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 entirety.

[0526] Example 28

[0527] In another exemplary manner, 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 is further characterized as illustrated and described below herein.

[0528] The present invention is characterized by a method for screening one or more potential drug candidate compounds to determine whether they are therapeutically effective, using 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 an experiment; (b) contacting at least one of cells, body fluids, tissues, organs, or animals with at least one of the reference compound, potential drug candidate compound, or negative control compound; (c) parallel determining one or more drug parameters or characteristics from contacting cells, body fluids, tissues, organs, or animals with at least one of the reference compound, potential drug candidate compound, or negative control compound; and (d) determining whether one or more drug parameters or characteristics of the negative control, reference compound, or drug candidate compound are 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 cell assays, colchicine binding site cell assays, reversibility assays, kinetic solubility assays, microsome stability assays, plasma protein binding studies, cell permeability studies, or cell cycle analysis by flow cytometry. In another non-limiting example, an ex vivo model may include a resected tumor in a chicken egg model. In yet another non-limiting example, an in vivo model may include pharmacokinetic (PK) studies, brain pharmacokinetic (PK) studies, maximal tolerance studies, efficacy studies, metabolite profiling in hepatocytes, survivability studies, or tumor size studies.

[0530] In some non-limiting examples, the drug parameters or characteristics analyzed may 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 cases, cells may be contacted with reference compounds, potential drug candidate compounds, or negative control compounds. These cells may include primary tumor cells, cancer cells, U87 glioblastoma cells, LN-18 glioblastoma cells, HMC3 microglia cells, patient-derived GBM cells, MDR1-MDCK (multidrug-resistant-1-Mandin-Darby canine kidney) cells, human cryopreserved hepatocytes, or cells used in patient-derived xenograft models. In other non-limiting cases, bodily fluids may be contacted with reference compounds, potential drug candidate compounds, or negative control compounds. These fluids may include plasma, cerebrospinal fluid, brain homogenates, urine, whole blood, serum, or tumor homogenates. In some non-limiting cases, tissues may be contacted with reference compounds, potential drug candidate compounds, or negative control compounds. These tissues may include brain tumor tissue, normal brain tissue, tumor tissue, or solid tumor tissue. In other non-limiting cases, organs may be contacted with reference compounds, potential drug candidate compounds, or negative control compounds. Organs may include the brain, mammary glands, skin, lungs, liver, bones, or connective tissue. In other non-limiting examples, animals may be brought into contact with a reference compound, a potential drug candidate compound, or a negative control compound. Animals may include mice, rats, dogs, monkeys, rabbits, or pigs.

[0532] Example 29

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

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

[0535] A 60-year-old man visits his doctor complaining of weakness and paralysis 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 doctor takes a thorough medical history, performs a neurological examination, and orders a brain MRI, including contrast, to determine the underlying cause of the man's symptoms. The man's examination results reveal that he has brain cancer suspected to be glioblastoma. Brain biopsy confirms the glioblastoma. To treat the brain cancer, the doctor prescribes an oral composition containing the RGN6024 compound, as described herein, in the form of tablets to be taken once daily at a dose of 1 mg / kg. The man adheres to the prescribed regimen, and his symptoms gradually improve with each dose over the next month. The nausea the patient experiences is mild and well controlled with antiemetics. A follow-up MRI four weeks after treatment shows an overall reduction in the size of the glioblastoma.

[0536] Example 29.2 Subcutaneous administration for the treatment of brain cancer (glioblastoma)

[0537] A 60-year-old man visits his doctor complaining of weakness and paralysis on the right si...

Claims

1. A compound according to formula (I-D), 【Chemistry 1】 In the formula, R ５ but, 【Chemistry 2】 【Transformation 3】 Selected from, R ６ It is one of the following: 【Chemistry 4】 【Transformation 5】 , compound.

2. The aforementioned compound, 【Transformation 6】 The compound according to claim 1.

3. A composition comprising at least one compound as described in claim 1.

Images