10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6-dihydroxy-4,5,6,7- tetrahydro-1H-spiro[benzo[B][1,5,4]oxathiazocine-3,1'-cyclopropane] 2,2-dioxide stereoisomers, compositions including the same, and methods of using the same

Abstract

Described herein are 10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6- dihydroxy-4,5,6,7-tetrahydro-1H-spiro[benzo[b][1,5,4]oxathiazepine-3,1'- cyclopropane] 2,2-dioxide stereoisomer compounds and pharmaceutical compositions thereof. Also provided are methods of using the same to treat, prevent, or ameliorate diseases or disorders in which the RAS pathway is upregulated, such as cancer, RASopathies, and laminopathies.

Description

[0001] 1. Cross-references to related applications This application claims the benefits of U.S. Provisional Application No. 63 / 722,523, filed November 19, 2024, and U.S. Provisional Application No. 63 / 618,167, filed January 5, 2024, the disclosures of which are incorporated herein by reference in their entirety. 2. Technical Field This document provides stereoisomers of 10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6-dihydroxy-4,5,6,7-tetrahydro-1H-spiro[benzo[b][1,5,4]oxathiazazecyclooctene-3,1'-cyclopropane]2,2-dioxide. This document also provides pharmaceutical compositions comprising such compounds and methods of using such compounds to treat, prevent, and manage various conditions. 3. Background Technology Mitogen-activated protein kinase (MAPK) is associated with many cancers. MAPK specifically phosphorylates serine / threonine residues of proteins, which are activated by various external stimuli (e.g., mitogens and growth factors) to exert their effects within the cell. MAPK activation regulates many physiologically significant cellular functions, such as cell growth, survival, apoptosis, differentiation, proliferation, and gene expression. (G. Pearson et al., Endocr: Rev (, 2001, pp. 153-183.) MEK1 and MEK2 are two human kinases in the middle of the MAPK cascade, involving upstream rat sarcoma virus (RAS)-rapidly accelerated fibrosarcoma (RAF) and downstream extracellular regulated kinase (ERK). This signaling cascade leading to ERK phosphorylation has been extensively studied in cancer pathology. Human cancers often carry mutations in members of the RAS oncogene family, which drive tumorigenesis by increasing cell proliferation and survival. These members are small protein GTPases regulated by a switch between an active GTP-linked state and an inactive GDP-binding state, controlled by a complex network of guanine exchange factor (GEF, which favors RAS-GTP) and GTPase activator (GAP, which favors RAS-GDP). (Cox AD et al., Nat Rev Drug Discov2014;13:828-51. RAS activation due to external recruitment or intrinsic mutations of transmembrane tyrosine kinase receptors propagates via downstream RAF-MEK-ERK and PI3K-AKT signaling pathways. In addition to mutations conferring RAS activation independent of physiological regulators, human cancers carry mutations in other RAS network genes such as NF1 (encoding neurofibroma protein, a RAS GAP), BRAF, or PTPN11 (encoding SHP2 tyrosine phosphatase involved in RAS activation). (Decroocq J et al., 2014;13:828-51.) Leukemia (2022) 36:1237-1252. Several non-ATP-competitive, allotropic MEK1 / 2 inhibitors have been developed and evaluated in clinical trials, primarily for cancers with aberrant ERK1 / 2 signaling activation, and four of these are FDA-approved drugs. (Zhao Y, Adjei AA) Nat Rev Clin Oncol . 2014;11:385;Caunt CJ, Sale MJ, Smith PD, Cook SJ. Nat Rev Cancer 2015;15:577;Shang J, Lu S, Jiang Y, Zhang J. Chem Biol Drug Des. 2016;88:485; Hao C et al., Eu. J. Med. Chem. 2023, 251: 115236; Heinzerling L., ESMO Open 2019;4:e000491.doi:10.1136 / esmoopen-2019-000491;Stalnecker CA et al., Sci Signal. 2020, 13(624): 1-14;Echevarria-Vargas IM et al., Melanoma Manag.(2017) 4(4), 183-186;Wang A et al., IUBMB Life, 2013, 65(9):748-758。). These inhibitors are selective for MEK1 / 2 because they bind to non-ATP-competitive sex loci. In addition to cancer, MEK1 / 2 inhibitors have the potential to be used for other diseases.

[0004] The compound, chemically named 10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6-dihydroxy-4,5,6,7-tetrahydro-1H-spiro[benzo[b][1,5,4]oxathiazazepine-3,1'-cyclopropane]2,2-dioxide, belongs to a class of novel MEK1 / 2 inhibitors with a macrocyclic scaffold, disclosed in US 9,034,861, which is incorporated herein by reference in its entirety. 4. Summary of the Invention This article provides pure compound 1, or its pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug: 1 This article also provides pure compound 2, or its pharmaceutically acceptable salts, solvates, hydrates, tautomers, esters, isotopes, or prodrugs: 2 This article also provides pure compound 3, or its pharmaceutically acceptable salts, solvates, hydrates, tautomers, esters, isotopes, or prodrugs: 3 This article also provides pure compound 4, or its pharmaceutically acceptable salts, solvates, hydrates, tautomers, esters, isotopes, or prodrugs: 4 Also provided are pharmaceutical compositions formulated for administration via appropriate routes and methods, containing effective concentrations of compound 1, compound 2, compound 3 or compound 4 provided herein, and optionally comprising at least one pharmaceutical carrier.

[0006] In one embodiment, delivery of the pharmaceutical composition can effectively treat levels of diseases or conditions upregulated by the RAS pathway (e.g., cancer, RAS protein disorders (e.g., Noonan syndrome, Costello syndrome, etc.), and laminopathies (e.g., cardiomyopathy, muscular dystrophy, developmental dysplasia, etc.)). In one embodiment, delivery of the pharmaceutical composition can effectively treat and / or prevent levels of cancer, RAS protein disorders, and laminopathies. In one embodiment, delivery of the pharmaceutical composition can effectively improve levels of cancer, RAS protein disorders, and laminopathies.

[0007] In one embodiment, this document provides a method for treating diseases or conditions with upregulated RAS pathways (e.g., cancer, RAS proteinopathies, and laminopathies), said method comprising administering the compounds or pharmaceutical compositions provided herein. This document also provides combination therapies using the compounds or pharmaceutical compositions provided herein in combination with a therapy (e.g., another pharmaceutical agent having activity against diseases or conditions with upregulated RAS pathways (e.g., cancer, RAS proteinopathies, and laminopathies) or their symptoms). Examples of therapies within the scope of said methods include, but are not limited to, surgery, chemotherapy, radiation therapy, biotherapy, stem cell transplantation, cell therapy, and combinations thereof.

[0008] These and other aspects of the subject matter described herein will become clear when the following detailed description is consulted. 5. Description of the attached drawings Figure 1 Provide the PK plasma curve of compound 1 on day 1 when tested in a clinical trial setting.

[0010] Figure 2 Provide the PK plasma curve of compound 1 on day 22 when tested in a clinical trial setting. 6. Detailed Implementation 6.1 Definition Unless the context clearly indicates otherwise, as used herein, in this specification, and in the appended claims, the indefinite articles “a” and “an” and the definite article “the” include both a plurality and a single indicator.

[0012] As used herein, the terms “comprising” and “including” are used interchangeably. The terms “comprising” and “including” should be interpreted as specifying the presence of the mentioned stated features or components, but do not preclude the presence or addition of one or more features, components, or groups thereof. Additionally, the terms “comprising” and “including” are intended to include instances covered by the term “consisting of”. Therefore, the term “consisting of” can be used in place of the terms “comprising” and “including” to provide more specific embodiments of the invention.

[0013] The term "composed of" means that the subject matter has at least 90%, 95%, 97%, 98%, or 99% of the stated features or components that constitute the subject matter. In another embodiment, the term "composed of" excludes any other features or components from any subsequent enumeration, except those features or components that are not essential for the technical effect to be achieved.

[0014] As used herein, the term "or" should be interpreted as an inclusive "or" of any one or any combination. Therefore, "A, B, or C" means any of the following: "A; B; C; A and B; A and C; B and C; A, B, and C". Exceptions to this definition will only occur if a combination of elements, functions, steps, or actions is inherently mutually exclusive in some way.

[0015] As used herein, the terms "about" or "approximately" mean an acceptable error for a particular value as determined by those skilled in the art, depending in part on how the value was measured or determined. In some embodiments, the terms "about" or "approximately" mean within 1, 2, 3, or 4 standard deviations. In some embodiments, the terms "about" or "approximately" mean within 50%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.05% of a given value or range.

[0016] As used herein and unless otherwise indicated, "substantially free of" another chemical compound or composition means, in some embodiments, that the compound or composition contains less than about 50 wt%, 45 wt%, 40 wt%, 35 wt%, 30 wt%, 25 wt%, 20 wt%, 15 wt%, 10 wt%, 9 wt%, 8 wt%, 7 wt%, 6 wt%, 5 wt%, 4 wt%, 3 wt%, 2 wt%, 1 wt%, 0.5 wt%, 0.4 wt%, 0.3 wt%, 0.2 wt%, 0.1 wt%, 0.05 wt%, or 0.01 wt% of another chemical compound or composition.

[0017] As used herein and unless otherwise specified, a "substantially chemically pure" compound is substantially free of other chemical compounds (i.e., chemical impurities). In some embodiments, a substantially chemically pure compound contains, by weight, less than about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.05%, or 0.01% of one or more other chemical compounds. The detection of other chemical compounds can be performed by any method obvious to a person skilled in the art, including but not limited to chemical analysis methods such as mass spectrometry, spectral analysis, thermal analysis, elemental combustion analysis, and / or chromatographic analysis.

[0018] As used herein and unless otherwise indicated, the term "pure" when applied to chiral compounds or "stereoisomers" or "stereoisopure" means that one stereoisomer of the disclosed compound is substantially free of other stereoisomers of the compound. For example, a stereoisopure compound having one chiral center will substantially be free of its opposite enantiomers. A stereoisopure compound having two chiral centers will substantially be free of other diastereomers of the compound. Typical stereoisopure compounds contain more than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of other stereoisomers of the compound, more than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of other stereoisomers of the compound, more than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of other stereoisomers of the compound, or more than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of other stereoisomers of the compound. The compounds disclosed herein have a chiral center and can exist as individual enantiomers or diastereomers and mixtures thereof. All such isomers are included in the embodiments disclosed herein, including mixtures thereof.

[0019] The embodiments disclosed herein cover the use of the pure stereoisomeric forms of such compounds as well as the use of mixtures of those forms. For example, mixtures comprising equal or unequal amounts of enantiomers of a particular compound can be used in the methods and compositions disclosed herein. These isomers can be asymmetrically synthesized or resolved using standard techniques, such as chiral columns or chiral resolving agents. See, for example, Jacques, J. et al. Enantiomers, Racemates and Resolutions (Wiley-Interscience, New York, 1981); Wilen, SH et al., Tetrahedron 33:2725 (1977); Eliel, EL, Stereochemistry of Carbon Compounds (McGraw-Hill, NY,1962); and Wilen, SH, Tables of Resolving Agents and Optical Resolutions Page 268 (edited by EL Eliel, Univ. of Notre Dame Press, Notre Dame, IN, 1972).

[0020] As used herein and unless otherwise indicated, the term "prodrug" means a compound that can be hydrolyzed, oxidized, or otherwise reacted under biological conditions (in vitro or in vivo) to yield an active compound, particularly the compounds disclosed herein. Examples of prodrugs include, but are not limited to, derivatives and metabolites of the compounds disclosed herein, including biohydrolyzable moieties such as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable acylureas, and biohydrolyzable phosphate ester analogs. In some embodiments, the prodrug of a compound having a carboxyl functional group is a lower alkyl ester of a carboxylic acid. Carboxylic acid esters are conveniently formed by esterification of any carboxylic acid moiety present on the molecule. Prodrugs are generally prepared using well-known methods, such as... Burger's Medicinal Chemistry and Drug Discovery 6th edition (edited by Donald J. Abraham, 2001, Wiley) and Design and Application of Prodrugs The method described in (H. Bundgaard, ed., 1985, Harwood Academic Publishers Gmfh).

[0021] "Tautomers" refer to the isomeric forms of a compound that are in equilibrium with each other. The concentration of the isomeric forms will depend on the environment in which the compound exists and can vary depending on, for example, whether the compound is a solid or in an organic or aqueous solution. For example, in aqueous solution, pyrazole can exhibit the following isomeric forms, which are referred to as tautomers of each other: .

[0022] Unless otherwise specified, as used herein, the terms "solvent" and "solventization" refer to the solid form of a substance containing a solvent. The terms "hydrate" and "hydration" refer to a solvate in which the solvent comprises water. As used herein, the terms "solvent" and "solventization" may also refer to solvates of salts, eutectics, or molecular complexes. As used herein, the terms "hydrate" and "hydration" may also refer to hydrates of salts, eutectics, or molecular complexes.

[0023] The term "isotope" as used herein refers to a compound that may contain one or more atomic isotopes in a non-natural proportion. For example, isotopes of compounds 1 to 4 may contain radioactive isotopes (e.g., tritium) at one or more sites. 3 H) and / or carbon-14 ( 14 C) Radioactive labeling, or isotopic enrichment at one or more sites, such as enrichment of deuterium (C) 2 H), carbon-13 ( 13 C), Oxygen-18 ( 18 O) and / or nitrogen-15 (15 N). In some embodiments, the compound described herein may be used at one or more sites with a radioactive isotope (e.g., tritium). 3 H) and / or carbon-14 ( 14 C)) Radioactive labeling, and also isotopic enrichment at one or more sites, such as enrichment of deuterium ( 2 H), carbon-13 ( 13 C), Oxygen-18 ( 18 O) and / or nitrogen-15 ( 15 N).

[0024] All isotopic variants of compounds 1 to 4, whether or not radioactive, are intended to be covered within the scope of the embodiments provided in this invention. In some embodiments, isotopes of compounds 1 to 4 are provided, for example, compounds 1 to 4 enriched with deuterium, carbon-13, or nitrogen-15.

[0025] Regarding the compounds presented herein, when a particular atomic position is specified as having deuterium or "D", it should be understood that the deuterium abundance at that position is significantly greater than the natural abundance of deuterium (which is about 0.0156%). In certain implementations, for each designated deuterium atom, the location designated as having deuterium typically has the following minimum isotopic enrichment factors: at least 100 (1.56% deuterium inclusion), at least 500 (7.8% deuterium inclusion), at least 1000 (15.6% deuterium inclusion), at least 2000 (31.2% deuterium inclusion), at least 3000 (46.8% deuterium inclusion), at least 3500 (54.6% deuterium inclusion), at least 4000 (62.4% deuterium inclusion), at least 4500 (70.2% deuterium inclusion), at least 5000 (78% deuterium inclusion), at least 5500 (85.8% deuterium inclusion), at least 6000 (93.6% deuterium inclusion), at least 6089.7 (95% deuterium inclusion), and at least 6217.9. (97% deuterium inclusion), at least 6346.2 (99% deuterium inclusion) or at least 6378.2 (99.5% deuterium inclusion).

[0026] The isotope enrichment and isotope enrichment factors of the compounds presented herein can be determined using conventional analytical methods (including mass spectrometry and nuclear magnetic resonance spectroscopy) known to those skilled in the art.

[0027] As used herein and unless otherwise specified, the term "pharmaceutically acceptable salt" means a salt prepared from a pharmaceutically acceptable, relatively non-toxic acid (including inorganic and organic acids). In some embodiments, suitable acids include, but are not limited to, acetic acid, adipic acid, 4-aminosalicylic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, camphoric acid, camphorsulfonic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, carbonic acid, citric acid, cyclohexane, dihydrogen phosphate, 2,5-dihydroxybenzoic acid (gentianic acid), 1,2-ethanedisulfonic acid, ethanesulfonic acid, fumaric acid, galacturonic acid, gluconic acid, glucuronic acid, glutamic acid, glutamate, and glutaraldehyde. Acids, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, isobutyric acid, hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, monohydrocarbonic acid, monohydrophosphoric acid, monohydrosulfuric acid, viscous acid, 1,5-naphthalenedisulfonic acid, nicotinic acid, nitric acid, oxalic acid, primic acid, pantothenic acid, phosphoric acid, phthalic acid, propionic acid, pyroglutamic acid, salicylic acid, octanoic acid, succinic acid, sulfuric acid, tartaric acid, toluenesulfonic acid, etc. (see, for example, SM Berge et al.) J. Pharm. Sci., 66:1-19 (1977); and Handbook of Pharmaceutical Salts: Properties, Selection and Use (PH Stahl and CG Wermuth, eds. (2002), Wiley, Weinheim). In some embodiments, the suitable acid is a strong acid (e.g., pKa less than about 1), including but not limited to hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, pyridinesulfonic acid, or other substituted sulfonic acids. Also included are salts of other relatively non-toxic compounds with acidic characteristics (including amino acids such as aspartic acid, etc.) and other compounds (e.g., aspirin, ibuprofen, saccharin, etc.). Acid addition salts can be obtained by contacting the neutral form of the compound with a sufficient amount of the desired acid (pure acid or in a suitable solvent).

[0028] As used herein and unless otherwise indicated, the terms “treat,” “treating,” and “treatment” refer to the relief or reduction of the severity of symptoms associated with a disease or ailment (such as cancer) being treated.

[0029] The term "prevention" includes suppressing symptoms of a specific disease or condition (such as cancer). Generally, the term "prevention" refers to administering medication to patients at risk of cancer before symptoms appear.

[0030] As used herein and unless otherwise indicated, the term “management” encompasses the prevention of recurrence of a particular disease or condition (e.g., cancer) in patients, the prolongation of remission in patients with a disease or condition, the reduction of mortality in patients, and / or the maintenance of severity of symptoms associated with the managed disease or condition.

[0031] As used in this article, "subject" or "patient" is an animal, usually a mammal, including humans, such as a human patient.

[0032] As used herein and unless otherwise specified, the terms “therapeutic effective amount” and “effective amount” for a compound mean an amount sufficient to provide therapeutic benefit in the treatment, prevention, and / or management of a disease (e.g., cancer), or to delay or minimize one or more symptoms associated with the disease or condition to be treated. The terms “therapeutic effective amount” and “effective amount” may also encompass amounts that improve overall therapy, reduce or avoid symptoms or causes of a disease or condition, or enhance the therapeutic efficacy of another therapeutic agent.

[0033] The terms "co-administered" and "in combination with" include the simultaneous, concurrent, or sequential administration of one or more therapeutic agents (e.g., the compounds provided herein and another cancer agent or supportive care agent) without a specific time limit. In one embodiment, the agents are simultaneously present in cells or in the patient's body or exert their biological or therapeutic effects simultaneously. In one embodiment, the therapeutic agents are in the same composition or unit dosage form. In another embodiment, the therapeutic agents are in separate compositions or unit dosage forms.

[0034] The term "supportive care agent" refers to any substance that treats, prevents, or manages adverse effects resulting from treatment with compound 1 or its tautomers or pharmaceutically acceptable salts.

[0035] In the context of cancer, inhibition can be assessed in particular by: inhibition of disease progression, inhibition of tumor growth, reduction of primary tumors, relief of tumor-related symptoms, inhibition of tumor-secreting factors, delay of the onset of primary or secondary tumors, slowing of the development of primary or secondary tumors, reduction of the occurrence of primary or secondary tumors, slowing or reducing the severity of secondary effects of the disease, preventing tumor growth and tumor regression, prolonging time to progression (TTP), prolonging progression-free survival (PFS), and prolonging overall survival (OS). As used herein, OS means the time from the start of treatment until death from any cause. As used herein, TTP means the time from the start of treatment until tumor progression; TTP does not include death. In one embodiment, PFS means the time from the start of treatment until tumor progression or death. In one embodiment, PFS means the time from the first dose of the compound to the first occurrence of disease progression or death from any cause. In one embodiment, the PFS rate will be calculated using Kaplan-Meier estimates. Event-free survival (EFS) refers to the time from the start of treatment until any treatment failure (including disease progression, discontinuation of treatment for any reason, or death). In one embodiment, overall response rate (ORR) refers to the percentage of patients who achieve a response. In one embodiment, ORR refers to the sum of the percentages of patients who achieve a complete and partial response. In one embodiment, ORR refers to the percentage of patients who, according to the IMWG uniform response criteria, have an optimal response ≥ partial response (PR). In one embodiment, duration of response (DoR) is the time from achieving a response until relapse or disease progression. In one embodiment, DoR is the time from achieving a response ≥ partial response (PR) until relapse or disease progression. In one embodiment, DoR is the time from the first recorded response until the first recorded progressive disease or death. In one embodiment, DoR is the time from the first recorded response ≥ partial response (PR) until the first recorded progressive disease or death. In one embodiment, time to response (TTR) refers to the time from the first dose of the compound to the first recorded response. In one embodiment, TTR refers to the time from the first dose of the compound to the first recorded response ≥ partial response (PR). In extreme cases, complete inhibition is referred to herein as prophylaxis or chemoprevention. In this context, the term "prevention" includes complete prevention of the onset of clinically apparent cancer or prevention of the onset of cancer in its preclinical, apparent stages. This definition also aims to cover prevention of transformation into malignant cells or to halt or reverse the progression of preclinical cells to malignant cells. This includes preventative treatment for subjects at risk of developing cancer.

[0036] The term “RAS protein pathology” refers to a group of syndromes, also known as disorders or conditions, caused by genetic alterations that send signals through the Ras / mitogen-activated protein kinase (Ras / MAPK) pathway. Although there are different RAS protein pathologies (currently fewer than 10 conditions) and each syndrome has unique characteristics, some common features include heart defects, skin, bone, eye and muscle problems, short stature, learning difficulties, appearance differences, and an increased risk of benign and cancerous tumors. RAS protein pathologies include, but are not limited to, Noonan syndrome, Costello syndrome, cardiofacial-skin syndrome, Legius syndrome, capillary arteriovenous malformation syndrome, hereditary gingival fibromatosis, SYNGAP1 syndrome, Leopard syndrome, neurofibromatosis 1 (NF1), and neurofibromatosis 2 (NF2) (see National Cancer Institute: https: / / dceg.cancer.gov / research / what-we-study / rasopathies).

[0037] The term "laminopathy" refers to a group of rare genetic disorders caused by mutations in the gene encoding laminin. It is included in the more general term nuclear membrane disorders, which refers to diseases associated with nuclear membrane defects. Laminopathy and other nuclear membrane disorders present with a wide variety of clinical symptoms, including skeletal and / or cardiac dystrophy, lipid dystrophy and diabetes, developmental delays, skin or neuropathy, white matter dystrophy, and premature aging. Most of these symptoms appear after birth, typically during childhood or adolescence. However, some laminopathy can lead to early death, and mutations in the laminin B1 (LMNB1 gene) can be fatal before or at birth. Known types of laminopathies and other nuclear membrane diseases include, but are not limited to, atypical Werner syndrome, Buschke-Ollendorff syndrome, cardiomyopathy, Charcot-Marie-Tooth disease, Emery-Dreifuss muscular dystrophies, Greenberg dysplasia, Hutchinson-Gilford progeria syndrome (HGPS), adult-onset autosomal dominant demyelinating leukodystrophy (ADLD), limb-girdle muscular dystrophy type 1B (LGMD1B), lipodystrophy with diabetes, hepatic steatosis, hypertrophic cardiomyopathy and leukodermoid melanocytic papules (LDHCP), mandibular acrodysplasia, Pelger-Huet anomaly (PHA), and restrictive dermatitis.

[0038] The term “neurofibromatosis” or “neurofibromatoses” refers to a group of genetic disorders belonging to the RAS protein lesions mentioned above, which result in the formation of tumors in nerve tissue. These tumors can occur anywhere in the nervous system, including the brain, spinal cord, and nerves. There are three types of neurofibromatosis: neurofibromatosis 1 (NF1), neurofibromatosis 2 (NF2), and schwannomas. NF1 is usually diagnosed in childhood, while NF2 and schwannomas are usually diagnosed in early adulthood. The tumors in these conditions are usually non-cancerous (benign), but can sometimes become cancerous (malignant). Symptoms are usually mild. However, complications of neurofibromatosis can include hearing loss, learning impairment, heart and blood vessel (cardiovascular) problems, vision loss, and severe pain. In some implementations, subjects diagnosed with NF1 have cutaneous and / or plexiform neurofibromas.

[0039] Unless otherwise specified, in the event of a difference between the illustrated chemical structure and the chemical name of the compound provided herein, the chemical structure shall prevail.

[0040] compound In some embodiments, compound 1 is provided herein. In some embodiments, compound 1, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof, is provided herein: 1 In some embodiments, compound 2 is provided herein. In some embodiments, compound 2, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof, is provided herein: 2 In some embodiments, compound 3 is provided herein. In some embodiments, compound 3, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof, is provided herein: 3 In some embodiments, compound 4 is provided herein. In some embodiments, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof, is provided herein: 4 In some embodiments, the compounds disclosed herein (e.g., compound 1, compound 2, compound 3, or compound 4) are stereoisomerically pure compounds or pharmaceutically acceptable salts, solvates, hydrates, tautomers, esters, isotopes, or prodrugs thereof. In some embodiments, the compounds according to compounds 1 to 4 are stereoisomerically pure compounds or pharmaceutically acceptable salts thereof. In some embodiments, the stereoisomeric pure compound comprises at least about 80% by weight of the specified stereoisomer and at most about 20% by weight of other stereoisomers, at least about 90% by weight of the specified stereoisomer and at most about 10% by weight of other stereoisomers, at least about 95% by weight of the specified stereoisomer and at most about 5% by weight of other stereoisomers, at least about 96.6% by weight of the specified stereoisomer and at most about 3.4% by weight of other stereoisomers, at least about 97% by weight of the specified stereoisomer and at most about 3% by weight of other stereoisomers, at least about 99% by weight of the specified stereoisomer and at most about 1% by weight of other stereoisomers, or at least about 99.9% by weight of the specified stereoisomer and at most about 0.1% by weight of other stereoisomers. In some embodiments, the weight is based on the total weight of the compound.

[0041] 6.2 Methods for preparing compounds This article also provides methods for preparing the compounds disclosed herein.

[0042] In one embodiment, compound 1 provided herein can be prepared using conventional organic synthesis and commercially available starting materials. For example, and without limitation, compound 1 can be prepared as outlined in Schemes 1 and 1a below. It should be noted that those skilled in the art know how to modify the procedures described in the illustrative schemes to obtain the desired product.

[0043] Option 1 It should be noted that the acetone protecting group in Scheme 1 can be replaced by any other protecting group known in the art that is suitable for diols.

[0044] Option 1a Option 1a: Synthetic intermediate A: It should be understood that the benzyl protecting group in Scheme 1a may be replaced by any other protecting group known in the art that is suitable for phenol groups.

[0045] In one embodiment, compound 4 can be prepared using L-erythrose instead of D-erythrose according to schemes 1 and 1a.

[0046] In one embodiment, compounds 2 and 3 provided herein can be prepared using conventional organic synthesis and commercially available starting materials. For example, and without limitation, compounds 2 and 3 can be prepared as outlined in Scheme 2 below. It should be noted that those skilled in the art know how to modify the procedures described in the illustrative scheme to obtain the desired products.

[0047] Option 2 6.3 Usage Method In one embodiment, this document provides a method for treating diseases or conditions with upregulated RAS pathways (e.g., cancer, RAS proteinopathy, and laminopathy), the method comprising administering to a patient a compound disclosed herein (e.g., compound 1, compound 2, compound 3, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof). In one embodiment, this document provides a method for treating cancer. In one embodiment, this document provides a method for treating diseases or conditions with upregulated RAS pathways (e.g., cancer, RAS proteinopathy, and laminopathy) by administering the compound to a patient. In one embodiment, this document provides a method of using a compound disclosed herein (e.g., compound 1, compound 2, compound 3, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof) for treating cancer.

[0048] In one embodiment, this document provides a method for preventing diseases or conditions (e.g., cancer, RAS proteinopathy, and laminopathy) that are upregulated by the RAS pathway, the method comprising administering to a patient a compound provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof). In one embodiment, this document provides a method for preventing cancer. In one embodiment, this document provides a method for preventing diseases or conditions (e.g., cancer, RAS proteinopathy, and laminopathy) that are upregulated by the RAS pathway, wherein the method comprises administering the compound to a patient. In one embodiment, this document provides a method of using a compound provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof) for cancer prevention.

[0049] In one embodiment, this document provides a method for managing diseases or conditions with upregulated RAS pathways (e.g., cancer, RAS proteinopathy, and laminopathy), the method comprising administering to a patient a compound provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof). In one embodiment, this document provides a method for managing cancer. In one embodiment, this document provides a method for managing diseases or conditions with upregulated RAS pathways (e.g., cancer, RAS proteinopathy, and laminopathy), wherein the method comprises administering the compound to a patient. In one embodiment, this document provides compounds provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or pharmaceutically acceptable salts, solvates, hydrates, tautomers, esters, isotopes, or prodrugs thereof) for use in methods of managing cancer.

[0050] In some implementations, RAS protein lesions include, but are not limited to, neurofibromatosis 1 (NF1), neurofibromatosis 2 (NF2), Noonan syndrome, Costello syndrome, cardiofacial-skin syndrome, Lex syndrome, capillary arteriovenous malformation syndrome, hereditary gingival fibromatosis, or SYNGAP1 syndrome. In some implementations, NF1 is associated with cutaneous and / or plexiform neurofibromas.

[0051] In some implementations, laminopathy includes, but is not limited to, atypical Werner syndrome, Buschk-Olendorf syndrome, cardiomyopathy, Charcot-Marie-Tuss disease, Emory-Dreyfus muscular dystrophy, Greenberg dysplasia, Hutchinson-Gilford progeria syndrome (HGPS), adult-onset autosomal dominant demyelinating leukodystrophy (ADLD), limb-girdle muscular dystrophy type 1B (LGMD1B), lipodystrophy with diabetes, hepatic steatosis, hypertrophic cardiomyopathy and leukodermoid melanocytic papules (LDHCP), acrodysplasia of the mandible, Perger-Schutter abnormality (PHA), or restrictive dermatitis.

[0052] In one specific embodiment of the method described herein, the disease or condition is a neurofibromatosis (NF)-related condition. In some embodiments, neurofibromatosis is neurofibromatosis 1 (NF1), neurofibromatosis 2 (NF2), or schwannomatosis. In some such embodiments, the disease or condition is cancer. In one specific embodiment of the method described herein, the disease or condition is cancer associated with neurofibromatosis type 1 (NF1). In one specific embodiment, this document provides a method for treating NF1 in a subject with NF1. In some embodiments, the method provided herein includes treating cutaneous and / or plexiform neurofibromas in NF1 in a subject with NF1. In one embodiment of the method described herein, the disease or condition is cancer associated with neurofibromatosis type 2 (NF2). In one embodiment of the method described herein, the disease or condition is schwannomatosis. In one embodiment of the method described herein, the disease or condition is cancer associated with schwannomatosis.

[0053] In other respects, this document provides a method for treating, preventing, or prophylaxis cancer, the method comprising administering to a subject in need an effective amount of a compound provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof). In some embodiments, the cancer may be selected from brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, gastric cancer, prostate cancer, kidney cancer, colorectal cancer, melanoma, or leukemia. In other or additional embodiments, the cancer is brain cancer or adrenocortical cancer. In other or additional embodiments, the cancer is breast cancer. In other or additional embodiments, the cancer is ovarian cancer. In other or additional embodiments, the cancer is pancreatic cancer. In other or additional embodiments, the cancer is gastric cancer. In other or additional embodiments, the cancer is prostate cancer. In other or additional embodiments, the cancer is kidney cancer. In other or additional embodiments, the cancer is colorectal cancer. In other or additional embodiments, the cancer is myeloid leukemia. In other or additional embodiments, the cancer is glioblastoma. In other or additional embodiments, the cancer is follicular lymphoma. In other or additional embodiments, the cancer is pre-B-cell acute leukemia. In other or additional embodiments, the cancer is chronic lymphocytic B-cell leukemia. In other or additional embodiments, the cancer is mesothelioma. In other or additional embodiments, the cancer is small cell lung cancer. In one specific embodiment, the cancer is low-grade severe ovarian cancer.

[0054] In some embodiments, this document provides a method for inhibiting the proliferation of cells with RAS mutations, the method comprising administering a compound provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof). In some embodiments, cancer is associated with RAS mutations. In some embodiments, this document provides a method for inducing apoptosis in cells with RAS mutations, the method comprising administering a compound provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof). In some embodiments, this document provides a method for inhibiting the proliferation of cells with KRAS mutations, the method comprising administering a compound provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof). In some embodiments, cancer is associated with KRAS mutations. In some embodiments, this document provides a method for inducing apoptosis in cells with KRAS mutations, the method comprising administering compounds described herein (e.g., compound 1, compound 2, compound 3, compound 4, or pharmaceutically acceptable salts, solvates, hydrates, tautomers, esters, isotopes, or prodrugs thereof). In some embodiments, this document provides a method for inhibiting the proliferation of cells with NRAS mutations, the method comprising administering compounds described herein (e.g., compound 1, compound 2, compound 3, compound 4, or pharmaceutically acceptable salts, solvates, hydrates, tautomers, esters, isotopes, or prodrugs thereof). In some embodiments, cancer is associated with NRAS mutations. In some embodiments, this document provides a method for inducing apoptosis in cells with RAS mutations, the method comprising administering compounds described herein (e.g., compound 1, compound 2, compound 3, compound 4, or pharmaceutically acceptable salts, solvates, hydrates, tautomers, esters, isotopes, or prodrugs thereof). In some embodiments, the KRAS mutation is located at codons 12, 13, 59, 61, and / or 146. In some implementations, mutant forms of the KRAS protein have one or more amino acid substitutions selected from the group consisting of: G12C, G12S, G12R, G12F, G12L, G12N, G12A, G12D, G12V, G13C, G13S, G13D, G13V, G13P, S17G, P34S, A59E, A59G, A59T, Q61K, Q61L, Q61R, and Q61H.In some implementations, the mutant form of the KRAS protein has one or more amino acid substitutions selected from the group consisting of: G12C, G12R, G12S, G12A, G12D, G12V, G13C, G13R, G13S, G13A, G13D, G13V, A59E, A59G, A59T, Q61K, Q61L, Q61R, Q61H, K117N, K117R, K117E, A146P, A146T, and A146V.

[0055] In some embodiments, the RAS mutation is an HRAS or MRAS mutation. In one embodiment, the mutant form of the HRAS protein has one or more amino acid substitutions selected from the group consisting of: G12C, G12D, G12F, G12N, G12S, G12V, G13C, G13D, G13E, G13R, G13S, G13V, Q61H, Q61K, Q61L, and Q61R.

[0056] In one embodiment, the mutant form of the MRAS protein has one or more amino acid substitutions selected from G23V and T68I.

[0057] In some embodiments, this document provides a method for treating cancer in patients with RAF mutations, the method comprising administering a compound provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof). In some embodiments, this document provides a method for inhibiting the proliferation of cells with RAF mutations, the method comprising administering a compound provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof). In some embodiments, the cancer is associated with a RAF mutation. In some embodiments, the BRAF mutation is a K601E mutation. In some embodiments, the BRAF mutation is a V600E mutation.

[0058] In some embodiments, this document provides a method for treating cancer in patients with NF1 mutations, the method comprising administering a compound provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof).

[0059] In some embodiments, the cancer is resistant to treatment with one or more MEK protein kinase inhibitors. In other embodiments, the cancer is resistant to treatment with one or more RAF protein kinase inhibitors. In some embodiments, the cancer is resistant to BRAF / MEK inhibition. In some such embodiments, the cancer is an advanced solid tumor. In some embodiments, the cancer is associated with RAS, RAF, and / or NF1 mutations. In some embodiments, the BRAF mutation is a K601E mutation. In some embodiments, the BRAF mutation is a V600E mutation. In other embodiments, the resistance is acquired resistance. In other embodiments, the resistance is primary resistance. In other or additional embodiments, the cancer is resistant to anticancer agents.

[0060] In some embodiments, the compounds provided herein may be administered once daily (QD or qd) or divided into multiple daily doses. Additionally, administration may be continuous (i.e., administered daily for several consecutive days, or daily) or intermittent (e.g., periodic administration, i.e., including periods of rest for several days, weeks, or months). As used herein, the term "daily" is intended to mean the administration of a therapeutic compound once or more daily, for example, for a period of time.

[0061] In some embodiments, the administration of the compounds or pharmaceutical compositions disclosed herein may be between about 0.001 mg / kg body weight and about 100 mg / kg body weight / day (administered in a single or split dose), more preferably at least about 0.1 mg / kg body weight / day. Specific therapeutic doses may include, for example, about 0.01 mg to about 7000 mg of the compound, and preferably include, for example, about 0.05 mg to about 2500 mg of the compound. Depending on the specific application, the amount of active compound in a unit dose formulation may vary or be adjusted from about 0.1 mg to 1000 mg, preferably from about 1 mg to 300 mg, more preferably from 10 mg to 200 mg. In some cases, dose levels below the lower limit of the aforementioned range may be sufficient, while in other cases, larger doses may be used without causing any harmful side effects, for example by dividing such larger doses into several smaller doses administered throughout the day. The amount administered will be based on the specific IC50 of the compound used. 50 The value varies. In combination therapy where the compound is not the sole treatment, a smaller amount of the compound can be administered while still having a therapeutic or preventative effect.

[0062] In one embodiment, the compounds or pharmaceutical compositions disclosed herein are administered in the following dosage amounts: about 1 mg / day, 2 mg / day, 4 mg / day, 5 mg / day, 8 mg / day, 10 mg / day, 15 mg / day, 20 mg / day, 22 mg / day, 30 mg / day, 35 mg / day, 37 mg / day, 45 mg / day, 50 mg / day, 70 mg / day, 100 mg / day, 125 mg / day, 140 mg / day, 175 mg / day, 200 mg / day, 250 mg / day, 280 mg / day, 350 mg / day, 400 mg / day, 450 mg / day, 500 mg / day, 550 mg / day, 600 mg / day, 650 mg / day, 700 mg / day, 750 mg / day, 1000 mg / day, or 1400 mg / day. In one embodiment, the compounds or pharmaceutical compositions disclosed herein are administered in the following dosage amounts: about 2 mg / day, 4 mg / day, 8 mg / day, 15 mg / day, 22 mg / day, 30 mg / day, 37 mg / day, or 45 mg / day.

[0063] For clarity, it should be noted that, unless otherwise specified, the dosages of compounds mentioned herein refer to the amount of compound 1, compound 2, compound 3, or compound 4 in the form of a free base. In cases where, for example, a pharmaceutically acceptable salt of compound 1, 2, 3, or 4 is used, the amounts given above will need to be adjusted accordingly.

[0064] 6.4 Pharmaceutical Compositions Also provided are pharmaceutical compositions formulated for administration via appropriate routes and methods, comprising an effective concentration of compound 1, compound 2, compound 3 or compound 4 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope or prodrug thereof, and optionally comprising at least one pharmaceutical carrier.

[0065] The compounds can be formulated into suitable pharmaceutical preparations, such as solutions, suspensions, tablets, dispersible tablets, pills, capsules, powders, sustained-release formulations, or elixirs for oral administration, or sterile solutions or suspensions for ocular or parenteral administration, as well as transdermal patch formulations and dry powder inhalers. Typically, the compounds described above are formulated into pharmaceutical compositions using techniques and procedures well known in the art (see, for example, Ansel Introduction to Pharmaceutical Dosage Forms, 7th edition, 1999). In one specific embodiment, the compounds provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof) can be formulated into capsules. In some such embodiments, the compounds provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof) are formulated into 2 mg, 4 mg, or 15 mg capsules. In one specific embodiment, the compounds provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or pharmaceutically acceptable salts, solvates, hydrates, tautomers, esters, isotopes, or prodrugs thereof) are formulated into 15 mg capsules.

[0066] In the composition, one or more compounds or pharmaceutically acceptable salts at effective concentrations are mixed with a suitable drug carrier or mediator. In some embodiments, upon administration, the concentration of the compounds in the composition is such that it effectively delivers an amount sufficient to treat, prevent, or improve one or more symptoms and / or progression of a subject's condition (e.g., cancer).

[0067] Typically, the compositions are formulated for single-dose administration. To formulate the composition, a weight fraction of the compound is dissolved, suspended, dispersed, or otherwise mixed in a chosen medium at an effective concentration that would alleviate or improve the condition to be treated. Suitable drug carriers or media for administering the compounds provided herein include any such carriers known to those skilled in the art as being suitable for a particular mode of administration.

[0068] The amount of the active compound contained in a pharmaceutically acceptable carrier is sufficient to exert a therapeutically useful effect on the treated patient without any undesirable side effects. Therapeuticly effective concentrations can be determined empirically by testing the compound in the in vitro and in vivo systems described herein, and then extrapolated to the dosage for human use.

[0069] The concentration of the active compound in a pharmaceutical composition will depend on the rate of absorption, tissue distribution, inactivation, metabolism, and excretion of the active compound, the physicochemical characteristics of the compound, the dosing schedule and dosage, and other factors known to those skilled in the art. For example, the amount delivered may be sufficient to improve one or more symptoms of the cancer or disease disclosed herein.

[0070] The pharmaceutical composition may be in a unit dosage form suitable for a precise single-dose administration. The pharmaceutical composition will comprise a conventional pharmaceutical carrier or excipient and a compound according to the invention as the active ingredient. Furthermore, it may include other medical or pharmaceutical agents, carriers, adjuvants, etc.

[0071] Exemplary parenteral administration forms include solutions or suspensions of the active compound in sterile aqueous solutions (e.g., propylene glycol or dextrose solution). Such dosage forms may be appropriately buffered if necessary.

[0072] Suitable drug carriers include inert diluents or fillers, water, and various organic solvents. Where necessary, the drug composition may contain additional ingredients such as flavoring agents, binders, excipients, etc. Therefore, for oral administration, tablets containing various excipients (e.g., citric acid) can be used with various disintegrants (e.g., starch, alginate, and certain complex silicates) and binders (e.g., sucrose, gelatin, and gum arabic). Additionally, lubricants (e.g., magnesium stearate, sodium lauryl sulfate, and talc) are often used for tableting purposes. Similar types of solid compositions can also be used in soft and hard-filled gelatin capsules.

[0073] Therefore, preferred materials include lactose or milk sugar and high molecular weight polyethylene glycol. When the aqueous suspension or elixir is intended for oral administration, the active compound therein may be combined with various sweeteners or flavoring agents, coloring substances or dyes, and (if necessary) emulsifiers or suspending agents, as well as diluents (e.g., water, ethanol, propylene glycol, glycerin, or combinations thereof). Methods for preparing various pharmaceutical compositions having specific amounts of the active compound are known or obvious to those skilled in the art. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Ester, Pa., 18th edition (1990).

[0074] In some embodiments, the effective amount of the compounds provided herein (e.g., compound 1, compound 2, compound 3, compound 4, or pharmaceutically acceptable salts, solvates, hydrates, tautomers, esters, isotopes, or prodrugs thereof) in the pharmaceutical composition may be at a level that will exert the desired effect; for example, the unit dose for oral and parenteral administration is from about 0.001 mg / kg of subject body weight to about 100 mg / kg of subject body weight.

[0075] The dosage of the compound to be administered to the subject can vary widely and can be determined by a healthcare professional. In any given situation, the amount of compound administered will depend on factors such as the solubility of the active ingredient, the formulation used, and the route of administration. In one embodiment, applying a local concentration provides an intracellular exposure or concentration of about 0.01–10 mM.

[0076] Dosage can also be expressed in units other than mg / kg / day. For example, parenteral dosage can be expressed as mg / m³. 2 / day. Given the subject's height or weight, or both, those skilled in the art will readily understand how to convert the dose from mg / kg / day to mg / m². 2 / day. For example, a dose of 1 mg / kg / day for a 65 kg human is approximately equal to 38 mg / m². 2 / sky.

[0077] For convenience, the compounds disclosed herein (e.g., compound 1, compound 2, compound 3, compound 4, or their pharmaceutically acceptable salts, solvates, hydrates, tautomers, esters, isotopes, or prodrugs) may be administered orally. In one embodiment, when administered orally, the compound is given with food and water. In one embodiment, when administered orally, the compound is not given with food. In another embodiment, the compound is dispersed in water or fruit juice (e.g., apple juice or orange juice) and administered orally as a suspension.

[0078] The compounds disclosed herein (e.g., compound 1, compound 2, compound 3, compound 4, or pharmaceutically acceptable salts, solvates, hydrates, tautomers, esters, isotopes, or prodrugs thereof) may also be administered intradermally, intramuscularly, intraperitoneally, percutaneously, intravenously, subcutaneously, intranasally, epidurally, sublingually, intracerebrally, intravaginally, transdermally, rectally, transmucosally, by inhalation, or topically to the ear, nose, eye, or skin. The mode of administration is determined by a healthcare professional and may depend in part on the site of the medical condition.

[0079] The compounds described herein, or their pharmaceutically acceptable salts, solvates, hydrates, tautomers, esters, isotopes, or prodrugs, may be administered as the sole therapy. The compounds described herein, or their pharmaceutically acceptable salts, solvates, hydrates, tautomers, esters, isotopes, or prodrugs, may also be administered in combination with another therapy or multiple therapies.

[0080] Other therapies include, but are not limited to, the application of other therapeutic agents, radiation therapy, or both. Examples of therapies within the scope of the methods include, but are not limited to, surgery, chemotherapy, radiation therapy, biological therapy, stem cell transplantation, cell therapy, and combinations thereof.

[0081] When the compounds described herein are administered in combination with other therapeutic agents, the compounds do not need to be administered in the same pharmaceutical composition as the other therapeutic agents, and due to their different physical and chemical characteristics, they can be administered via different routes. For example, the compound or composition may be administered orally to produce and maintain a good blood level, while another therapeutic agent may be administered intravenously. Where possible, determining the mode of administration and its desirability within the same pharmaceutical composition is entirely within the knowledge of a skilled clinician. Initial administration may be performed according to protocols established in the art, and then the skilled clinician may modify the dosage, mode of administration, and timing of administration based on observed effects. The specific selection of the compound (and, where appropriate, other therapeutic agents and / or radiation) will depend on the attending physician's diagnosis and judgment of the patient's condition and the appropriate treatment regimen. Other therapeutic agents may include chemotherapeutic agents, such as antitumor substances. Such combination therapy may be achieved by means of simultaneous, sequential, or individual administration of the individual therapeutic components.

[0082] 7. Examples Example 1: Reference Synthesis of 10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6-dihydroxy-4,5,6,7-tetrahydro-1H-spiro[benzo[b][1,5,4]oxathiazazepine-3,1'-cyclopropane] 2,2-dioxide Step 1: Preparation of 3,4,5-trifluoro-l-nitrophenol 3,4,5-Trifluorophenol (14.81 g, 0.1 mol) was dissolved in glacial acetic acid (50 mL) and cooled to 4 °C. Concentrated nitric acid (5 mL, 70%) was added dropwise over 15 min, during which time the mixture turned yellow. After the addition of HNO3 was complete, the reaction mixture was heated to room temperature and stirred for another 30 min. TLC analysis of the aliquot extracted into ethyl acetate indicated the formation of new nonpolar spots and complete consumption of the starting material. The mixture was then diluted with ethyl acetate (200 mL), transferred to a separatory funnel, and thoroughly washed with water (3 x 100 mL). The organic layer was finally washed with brine, dried over anhydrous MgSO4, and evaporated under reduced pressure to give a crude product (17.3 g, 90%) that was a pale yellow oil. This crude material was used directly in subsequent reactions. 1 H NMR (400 MHz, CDCl3): 6.84 (m, 1H, ArH), 10.28 (brs, 1H, OH).

[0083] Step 2: Preparation of 3,4,5-trifluoro-2-nitro-phenylallyl ether K₂CO₃ (50 mmol) and allyl bromide (3.6 g, 30 mmol) were added to a solution of crude 3,4,5-trifluoro-2-nitrophenol (4.8 g, 25 mmol) in acetone (25 mL), and the mixture was heated to reflux for 2 h. TLC analysis of the reaction mixture (25% EtOAc:hexane) at this point revealed that all starting materials had been consumed and nonpolar spots were present. Heating was stopped, and the reaction mixture was allowed to cool. Most of the acetone was evaporated under vacuum, and the remaining residue was diluted with ether (50 mL) and washed sequentially with water. The organic ether layer was dried over MgSO₄ and concentrated under vacuum by rotary evaporation. The yellow-orange crude oil was further purified by silica gel rapid column chromatography using a hexane to 15% hexane:ethyl acetate gradient. The homogeneous fractions from the TLC were collected, combined, and evaporated under reduced pressure to give the allyl ether product (5.5 g, 95%). 1 H NMR (400 MHz, CDCl3): 4.65 (dt, J=1.6, 5.2 Hz, 2H, OCH2), 5.39 (d, J =12.0 Hz, 1H, =CH2), 5.45 (d, J=18.0 Hz, 1H, =CH2), 5.98 (m, 1H, =CH), 6.72 (m, 1H, ArH).

[0084] Step 3: Preparation of (3-allyloxy-5,6-difluoro-2-nitro-phenyl)-(2-fluoro-4-iodo-phenyl)-amine LHMDS solution (6.0 mL, 6.0 mmol, 1 M in THF) was added dropwise to a solution of 2-fluoro-4-iodophenylamine (1.1 g, 4.6 mmol) in 50 mL of THF at -78 °C. After stirring at -78 °C for 1 h, a solution of l-allyloxy-3,4,5-trifluoro-2-nitrobenzene (1.2 g, 5.1 mmol) in 10 mL of THF was added dropwise to the reaction mixture. The reaction mixture was stirred at -78 °C for another 1 h, then heated to room temperature and stirred for 16 h. 1 The reaction progress was monitored by ¹H NMR. After completion, the solvent was removed under reduced pressure. The obtained residue was dissolved in ethyl acetate, washed with water, dried over anhydrous Na₂SO₄, and concentrated. The residue was ground with hexane to give (3-allyloxy-5,6-difluoro-2-nitro-phenyl)-(2-fluoro-4-iodo-phenyl)-amine (900 mg) as a yellow solid. 1H-NMR (400 MHz, CDCl3): 4.62 (2H,d, J 4.8), 5.33-5.36 (1H, d, J=10), 5.48 (1H, d, J=17.2), 5.98-6.02 (1H, m),6.22 (1H, dd, J=2.4, 9.6), 6.36 (1H, dd, J=2, 10.4), 7.04-7.08 (1H, m), 7.45-7.52 (2H, m), 7.79 (1H, s).

[0085] Step 4: Preparation of 6-allyloxy-3,4-difluoro-N2-(2-fluoro-4-iodo-phenyl)-phenyl-1,2-diamine A suspension of (3-allyloxy-5,6-difluoro-2-nitro-phenyl)-(2-fluoro-4-iodo-phenyl)-amine (7, 0.9 g, 2 mmol) in ethanol (12 mL) was stirred at 70 °C to obtain a clear solution. A solution of freshly prepared Na₂S₂O₄ (1.04 g, 6 mmol) in water (2.5 mL) was added to this hot solution. The reaction mixture was stirred at 90 °C for 1 h. The progress of the reaction was monitored by TLC. After completion, the solvent was removed under reduced pressure. The residue was diluted with ethyl acetate, washed with water, and the organic phase was dried over anhydrous Na₂SO₄ and concentrated to give 6-allyloxy-3,4-difluoro-N₂-(2-fluoro-4-iodo-phenyl)-phenyl-1,2-diamine (730 mg) as a brown solid. 1H-NMR (400 MHz, CDCl3): 3.86 (2H, bs), 4.54 (2H, d, J=5.2), 5.34 (1H, d, J=10.8), 5.42 (1H, d, J=17.2), 5.66 (1H,bs), 6.02-6.09 (1H, m), 6.20 (1H, d, J=8.4), 6.62-6.66 (1H, m), 7.33 (1H, dd,J=2, 8.4), 7.63 (1H, d, J=2).

[0086] Step 5: Preparation of l-allyl-N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-allyloxyphenyl)cyclopropane-l-sulfonamide 3-(allyloxy)-5,6-difluoro-N-(2-fluoro-4-iodophenyl)benzene-1,2-diamine (420.2 mg, 1.0 mmol) was dissolved in anhydrous pyridine (1.0 mL), and l-allyl-cyclopropyl-1-sulfonyl chloride (250.0 mg, 1.38 mmol, freshly prepared) was added to this solution at room temperature. The mixture was heated in an oil bath under nitrogen for 48 h. TLC analysis of the mixture indicated the formation of new polar spots compared to the starting material. The reaction mixture was diluted with ethyl acetate and washed with 0.01 M HCl, water, and brine. The organic layer was dried over MgSO4 and concentrated under reduced pressure. A rapid chromatography of the crude material in 30% to 40% hexane:ethyl acetate was performed on silica gel to give a pure compound (375 mg, 66%). MS analysis: [M+H] + 565; 1H NMR (400 MHz, CDCl3): 0.81 (t, J=6.0 Hz, 2H, cyclopropyl-CH2), 1.26 (t, J=6.0 Hz, 2H, cyclopropyl-CH2), 2.73 (d, J=8.0 Hz, 2H, ----CH2), 4.62 (dt, J=1.2, 5.2 Hz, 2H, OCH2), 5.08 (dd, J=1.2, 16.0 Hz, 1H, =CH2), 5.13 (dt, J=1.6, 8.0 Hz, 1H, =CH2), 5.44 (dd, J=1.6, 8.0 Hz, 1H, =CH2), 5.51 (dt, J=1.6, 8.0 Hz, 1H, =CH2). 5.69(m, 1H, =CH), 6.07 (m, 1H, =CH), 6.12 (1H, s, NH), 6.44 (m, 1H, ArH), 6.56(dd, J=4.0, 12.0 Hz, 1H, ArH), 7.28 (d, J=8.0 Hz, 1H, ArH), 7.40 (dd, J=1.0,8.0 Hz, 2H, ArH).

[0087] Step 6: Preparation of (Z)-10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-4,7-dihydro-1H-spiro[benzo[b][1,5,4]oxathiazazepine-3,1'-cyclopropane] 2,2-dioxide Method A. At room temperature, Zhang catalyst [e.g.] Tetrahedron Letters[Prepared as described in 46 (2005) 7225-7228] A dilute solution of 1.5 mg / mL, 50 μL in CH2Cl2 was added to a CH2Cl2 solution (1.0 mL) of 1-allyl-N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-allyloxyphenyl)cyclopropane-1-sulfonamide (6.3 mg, 0.011 mmol) from step 5, and the mixture was stirred at room temperature for another 24 h. The mixture was then concentrated and purified by preparative TLC (silica gel), developed with hexane:ethyl acetate, and the bands corresponding to the new compound were collected and eluted with acetone. The desired compound (4.8 mg, 80%) was separated as a solid. MS analysis: [m+H] + 537; 1 H NMR (400 MHz, CDCl3): 0.74 (brs s, CH2), 1.14 (brs, CH2), 3.14 (m, 2H, CH2), 4.92 (s, 2H,OCH2), 5.46 (dd, J=12.0 Hz, 1H, =CH), 5.72 (dd, J=8.0, 12.0 Hz, 1H, =CH), 6.27 (s, 1H, NH), 6.51 (m, 2H, ArH), 7.18 (s, 1H, NH), 7.29 (d, J=8.0, 1H,ArH), 7.41 (d, J=12.0 Hz, 1H, ArH).

[0088] Method B. Hoveyda-Grubbs second-generation catalyst (120 mg, 0.19 mmol, 10 mol%) was added to a degassed solution of the diene (from step 5, 930 mg, 1.64 mmol) in dichloroethane (60 mL). The reaction mixture was stirred at 70 °C for 3 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by rapid column chromatography to give the desired compound (225 mg).

[0089] Step 7: Preparation of 10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6-dihydroxy-4,5,6,7-tetrahydro-1H-spiro[benzo[b][1,5,4]oxathiazazepine-3,1'-cyclopropane] 2,2-dioxide NMO (5.0 mg) was added via syringe to a THF solution (0.5 mL) of (E)-10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-4,7-dihydro-1H-spiro[benzo[b][1,5,4]oxathiazazepine-3,1'-cyclopropane]2,2-dioxide (4.8 mg, 0.009 mmol), followed by the addition of OsO4 solution (5.0 μL, 4% wt in water). The mixture was stirred overnight (14 h). TLC analysis showed complete consumption of the starting material, forming a highly polar product (50% hexane:ethyl acetate). The mixture was diluted with ethyl acetate (5.0 mL), washed with Na2S2O3 (1% solution, 2.0 mL), water, and finally brine. The organic layer was separated, dried over MgSO4, and evaporated. The crude compound was purified by preparative TLC, and the most polar band that moved through ethyl acetate was collected. Extraction of the collected silica bands with acetone yielded a racemic diol product (3.8 mg, 74%). MS analysis: [M+H] + 571; 1 H NMR (400 MHz, CDCl3): 0.68 (brs s, 2H,CH2), 0.75 (m, 1H), 1.17 (brs, 1H, CH2), 2.07 (s, 2H, CH2), 2.12 (s, 2H, CH2), 3.10-2.50 (m, 3H), 3.65 (m, 1H), 3.85 (d, 1H), 4.04 (brs, 1H), 4.42 (brt,1H), 6.40 (m, 1H), 6.88 (s, 1H, ArH), 7.28 (d, 1H, ArH), 7.30 (d, J=8.0, 1H,ArH).

[0090] Stereoisomers were obtained by separation using SFC under the following conditions: hexane:ethanol (90:10 v / v); column: Chiralcel OD-H (250 x 4.6 mm) 5 uM; flow rate: 1.5 ml / min; temperature: ambient; concentration: 1.0 mg / ml; UV detection: 220 nm.

[0091] Example 2: Enantioselective synthesis of compound 1 A. Preparation of intermediate A.

[0092] Step 1: Preparation of 3,4,5-trifluoro-2-nitrophenol 3,4,5-trifluorophenol was treated with nitric acid in acetic acid to give the title compound 3,4,5-trifluoro-2-nitrophenol.

[0093] Step 2: Preparation of 1-(benzyloxy)-3,4,5-trifluoro-2-nitrobenzene Treating 3,4,5-trifluoro-2-nitrophenol with potassium carbonate and benzyl bromide yields the title compound 1-(benzyloxy)-3,4,5-trifluoro-2-nitrophenol.

[0094] Step 3: Preparation of 3-(benzyloxy)-5,6-difluoro-N-(2-fluoro-4-iodophenyl)-2-nitroaniline The 1-(benzyloxy)-3,4,5-trifluoro-2-nitrobenzene from step 2 was treated with 2-fluoro-4-iodoaniline and bis(trimethylsilyl)aminolithium to give the title compound 3-(benzyloxy)-5,6-difluoro-N-(2-fluoro-4-iodophenyl)-2-nitrobenzene.

[0095] Step 4: Preparation of 3-(benzyloxy)-5,6-difluoro-N1-(2-fluoro-4-iodophenyl)benzene-1,2-diamine (Intermediate A) The 3-(benzyloxy)-5,6-difluoro-N-(2-fluoro-4-iodophenyl)-2-nitroaniline from step 3 was treated with ethanol:water containing sodium dithionite to give the title compound 3-(benzyloxy)-5,6-difluoro-N1-(2-fluoro-4-iodophenyl)benzene-1,2-diamine.

[0096] B. Preparation of compound 1 Step 1: Preparation of (2S,3S)-4-(benzyloxy)-2,3-dihydroxybutyraldehyde D-erythrose was treated with 2,2,2-trichloroethyliminosyl ester in trifluoromethanesulfonic acid to give the title compound (2S,3S)-4-(benzyloxy)-2,3-dihydroxybutanal.

[0097] Step 2: Preparation of (4S,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-carboxaldehyde The (2S,3S)-4-(benzyloxy)-2,3-dihydroxybutanal from step 1 was treated with acetone and camphor sulfonic acid to give the title compound (4S,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-carboxaldehyde.

[0098] Step 3: Preparation of ((4R,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methanol The (4S,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-carboxaldehyde from step 2 was treated with sodium borohydride in THF and heated to give the title compound ((4R,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methanol.

[0099] Step 4: Preparation of (4S,5S)-4-((benzyloxy)methyl)-5-(bromomethyl)-2,2-dimethyl-1,3-dioxolane Adding tetrabromomethane in a suitable solvent to ((4R,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methanol from step 3 yields the title compound (4S,5S)-4-((benzyloxy)methyl)-5-(bromomethyl)-2,2-dimethyl-1,3-dioxolane.

[0100] Step 5: Preparation of 1-(((4R,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methyl)cyclopropane-1-sulfonate butyl ester The (4S,5S)-4-((benzyloxy)methyl)-5-(bromomethyl)-2,2-dimethyl-1,3-dioxolane from step 4 was treated with butyl cyclopropane sulfonate and n-butyllithium at low temperature to give the title compound 1-(((4R,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methyl)cyclopropane-1-sulfonate butyl ester.

[0101] Step 6: Preparation of 1-(((4R,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methyl)cyclopropane-1-sulfonyl chloride The 1-(((4R,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methyl)cyclopropane-1-sulfonate butyl ester from step 5 was treated sequentially with potassium thiocyanate (KNCS) and thionyl chloride to give the title compound 1-(((4R,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methyl)cyclopropane-1-sulfonate chloride.

[0102] Step 7: Preparation of N-(6-(benzyloxy)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)phenyl)-1-(((4S,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methyl)cyclopropane-1-sulfonamide The intermediate A and a solution of 1-(((4R,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methyl)cyclopropane-1-sulfonyl chloride in pyridine were heated to 80 °C to give the title compound N-(6-(benzyloxy)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)phenyl)-1-(((4S,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methyl)cyclopropane-1-sulfonamide.

[0103] Step 8: Preparation of N-(3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-6-hydroxyphenyl)-1-(((4S,5S)-5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methyl)cyclopropane-1-sulfonamide The N-(6-(benzyloxy)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)phenyl)-1-(((4S,5S)-5-((benzyloxy)methyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methyl)cyclopropane-1-sulfonamide from step 7 was treated with palladium / carbon and hydrogen in a suitable solvent to give the title compound N-(3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-6-hydroxyphenyl)-1-(((4S,5S)-5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methyl)cyclopropane-1-sulfonamide.

[0104] Step 9: Preparation of (5R,6S)-10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6-dihydroxy-4,5,6,7-tetrahydro-1H-spiro[benzo[b][1,5,4]oxathiazazecyclooctene-3,1'-cyclopropane] 2,2-dioxide (Compound 1) The N-(3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-6-hydroxyphenyl)-1-(((4S,5S)-5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxolane-4-yl)methyl)cyclopropane-1-sulfonamide from step 8 was treated in THF with triphenylphosphine (Ph3P) and diethyl azodicarbonate (DEAD), followed by a deprotection step to give the title compound (5R,6S)-10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6-dihydroxy-4,5,6,7-tetrahydro-1H-spiro[benzo[b][1,5,4]oxazazezacyclooctene-3,1'-cyclopropane]2,2-dioxide (compound 1).

[0105] Example 3: Synthesis of Compound 2 and Compound 3 For the preparation of compounds 2 and 3, intermediate (E)-10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-4,7-dihydro-1H-spiro[benzo[b][1,5,4]oxathiazazecyclooctene-3,1'-cyclopropane]2,2-dioxide was prepared according to steps 1-6 of Example 1.

[0106] Step 1: Preparation of 7,8-difluoro-6-((2-fluoro-4-iodophenyl)amino)-1a,2,11,11a-tetrahydro-5H-spiro[benzo[b]epoxyvinyl[2,3-h][1,5,4]oxathiazacyclooctene-3,1'-cyclopropane]4,4-dioxide The (E)-10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-4,7-dihydro-1H-spiro[benzo[b][1,5,4]oxathiazacyclooctene-3,1'-cyclopropane] 2,2-dioxide from step 6 of Example 1 was treated with sodium periodate to give the title compound 7,8-difluoro-6-((2-fluoro-4-iodophenyl)amino)-1a,2,11,11a-tetrahydro-5H-spiro[benzo[b]epoxyvinyl[2,3-h][1,5,4]oxathiazacyclooctene-3,1'-cyclopropane] 4,4-dioxide in racemic mixture form.

[0107] Step 2: Preparation of 10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6-dihydroxy-4,5,6,7-tetrahydro-1H-spiro[benzo[b][1,5,4]oxathiazazepine-3,1'-cyclopropane] 2,2-dioxide The (1aS,11aR)-7,8-difluoro-6-((2-fluoro-4-iodophenyl)amino)-1a,2,11,11a-tetrahydro-5H-spiro[benzo[b]epoxyvinyl[2,3-h][1,5,4]oxathiazazecyclooctene-3,1'-cyclopropane] 4,4-dioxide from step 1 was treated with an aqueous sodium hydroxide solution to give the title compound (5S,6S)-10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6-dihydroxy-4,5,6,7-tetrahydro-1H-spiro[benzo[b][1,5,4]oxathiazazecyclooctene-3,1'-cyclopropane] 2,2-dioxide in the form of a racemic mixture of anti-diols.

[0108] Step 3: Preparation of (5S,6S)-10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6-dihydroxy-4,5,6,7-tetrahydro-1H-spiro[benzo[b][1,5,4]oxathiazazecyclooctene-3,1'-cyclopropane] 2,2-dioxide (compound 2) and (5R,6R)-10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6-dihydroxy-4,5,6,7-tetrahydro-1H-spiro[benzo[b][1,5,4]oxathiazazecyclooctene-3,1'-cyclopropane] 2,2-dioxide (compound 3) As described in steps 7a and 7b of Example 1, the racemic 10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6-dihydroxy-4,5,6,7-tetrahydro-1H-spiro[benzo[b][1,5,4]oxathiazazecyclooctene-3,1'-cyclopropane] 2,2-dioxide was separated by chiral HPLC to obtain the desired compound (5S,6S)-10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6-dihydroxy-4,5,6,7-tetrahydro-1H-spiro[benzo[b][1,5,4]oxathiazazecyclooctene-3,1'-cyclopropane] 2,2-dioxide (compound 2) and (5R,6R)-10,11-difluoro-12-((2-fluoro-4-iodophenyl)amino)-5,6-dihydroxy-4,5,6,7-tetrahydro-1H-spiro[benzo[b][1,5,4]oxathiazacyclooctene-3,1'-cyclopropane] 2,2-dioxide (compound 3).

[0109] Synthetic compound 4 According to steps 1-9 of Example 2, compound 4 was prepared by using L-erythrose instead of D-erythrose in step 1.

[0110] Example 4: In vitro cell proliferation assay HT29 cells obtained from ATCC were grown at 37°C in a humidified 5% CO2 incubator in DMEM medium supplemented with L-glutamine (Invitrogen) and 10% fetal bovine serum (Hyclone).

[0111] Cell proliferation was assessed by seeding 2,000 cells / well in 100 μL of DMEM / 10% FBS in 96-well plates and incubating overnight at 37°C in a humidified 5% CO2 incubator. The medium was replaced with fresh 100 μL of fresh DMEM / 10% FBS containing different concentrations of the compound. The compound was added in triplicate dilutions ranging from 3.3 μM to 4.5 nM. After incubation with the compound at 37°C in a humidified 5% CO2 incubator for 72 hours, cell viability was measured using a photometer after adding 100 μL / well of CellTiterGlo reagent (Promega). IC50 was calculated using SoftMax software. 50 .

[0112] IC50 assays were obtained for compound 1 and its enantiomer compound 4. 50 This indicates that compound 1 is 25 times more potent than its enantiomer, compound 4. Specifically, compound 1 exhibits IC... 50The value was 40 nM in biochemical assays, 16 nM in HT-29 proliferation assays, 15 nM in Colo-205 proliferation assays, and 27 nM in A-375 proliferation assays; while compound 4 showed an IC50 value. 50 The value was 191 nM in biochemical assays, 400 nM in HT-29 proliferation assays, 358 nM in Colo-205 proliferation assays, and 451 nM in A-375 proliferation assays. The reference compound RDEA119 (also known as refamatetinib, disclosed in U.S. Patent No. 7,759,518) is an acyclic diol compound that showed an IC50 value of 191 nM in biochemical assays. 50 The value is 21 nM and its enantiomers are approximately equivalent under the same conditions, IC 50 The value is 39 nM. A summary of the data can be found in Table 1.

[0113] Table 1: Summary of data comparisons between enantiomer forms Compound 4 is an enantiomer of compound 1. Therefore, the macrocyclic compounds disclosed herein exhibit improvements over those known in the art, not only because they possess a macrocyclic structure compared to the acyclic (linear) structures of known compounds, but also because of the enantiomeric differences in macrocyclic diols, which are not observed in acyclic (linear) diols. Specifically, as described above, compound 1 is 25 times more potent than its enantiomer, compound 4. This result is unexpected considering the lack of differences in the enantiomers of known acyclic diols in the art.

[0114] Example 5. Phase 1 clinical trial of compound 1 A phase 1 open-label study evaluating the safety, tolerability, pharmacokinetics, and pharmacodynamics of compound 1 in patients with advanced solid tumors driven by the MAPK pathway and who have RAS, NF1, or RAF mutations, or in patients whose BRAF / MEK inhibition has failed. The Phase 1 clinical trial was a multicenter, open-label, dose-escalation 3+3 study design to evaluate the safety, tolerability, pharmacokinetics (PK), pharmacodynamics (PD), and preliminary efficacy of compound 1 in patients with advanced solid tumors driven by the MAPK pathway and who have RAS, NF1, or RAF mutations, or in patients who have failed BRAF / MEK inhibition.

[0115] Compound 1 was tested in people with advanced solid tumors that have mutations in rat sarcoma virus (RAS), neurofibromatosis type I (NF1), or rapidly accelerating fibrosarcoma (RAF).

[0116] Patients with a histological or cytological diagnosis of advanced solid tumors driven by the mitogen-activated protein kinase (MAPK) pathway and possessing all of the following characteristics are eligible for inclusion: i. The tumor is inoperable. ii. The patient is unable or ineligible to receive standard care. iii. The patient has no available treatment options with known clinical benefit. iv. Documented evidence of mutations in rat sarcoma virus (RAS), neurofibromatosis type I (NF1), and / or rapidly accelerating fibrosarcoma (RAF). Patients with RAF mutations must have previously failed to inhibit the v-Raf murine sarcoma virus oncogene homolog B (BRAF) / MEK.

[0117] Patients in the study received a single oral dose of compound 1, followed by a one-week observation period, and then once daily for 28 days as one cycle throughout the study. Participants continued to take compound 1 daily for up to two years, or until: i. Their decision to withdraw from the research, or ii. They experienced unacceptable side effects, or iii. The progression of their disease, or another disease interfering with the use of the investigational drug, or iv. The initiator terminates the research.

[0118] In this study, compound 1 was presented in capsules of 1 mg, 4 mg, and 10 mg strengths, intended for once-daily oral administration. The doses were sequentially increased to 2 mg, 4 mg, 8 mg, 15 mg, 22 mg, 30 mg, 37 mg, and 45 mg.

[0119] In some implementations, the capsule formulation comprises compound 1 in solid form.

[0120] 1. Pharmacokinetic profile of compound 1 1.1 Interim Phase I Results PK data from the 2 mg and 4 mg cohorts in the first-in-human Phase 1 clinical trial of Compound 1 demonstrate a PK and safety profile that differentiates Compound 1 as a next-generation MEK inhibitor. Compound 1 achieved significant exposure and a favorable safety profile without adverse side effects such as rash or GI toxicity, which are typical symptoms of MEK inhibitors even at low doses. Its long half-life of approximately 70 hours and the ability to achieve a flat PK curve at steady state are designed to provide constant target inhibition while avoiding peak plasma toxicity, a unique PK profile for a MEK inhibitor intended to treat neurofibromatosis type 1 (NF1).

[0121] Table 5. Pharmacokinetic (PK) data for cohort 1 (2 mg) and cohort 2 (4 mg) at day 1 and day 22 (steady state). The data are geometric mean (geometric coefficient of variation (CV)%), Cmax: the highest concentration of the drug after the dose is administered; Cmin: the lowest concentration of the drug after the dose is administered; AUC: the area under the concentration-time curve and measures the total drug exposure (level) over the entire time.

[0122] 1.1.1 PK Results: • Plasma exposure increased with increasing dose and linear pharmacokinetic (PK) was observed; • A long half-life of approximately 70 hours will allow for once-daily dosing or longer intervals; • Prolonged systemic exposure with minimal fluctuations in plasma concentration of compound 1 at steady state (Cmax / Cmin ratio of 1.2) indicates the potential to achieve constant target inhibition. See also Figure 1 and Figure 2 .

[0123] At steady state, drug levels peaked at approximately 5 hours, with geometric mean maximum concentrations (Cmax) of 16.2 ng / mL and 61.3 ng / mL for the 2 mg and 4 mg dose groups, respectively. The mean elimination half-life was 67.9 hours, supporting once-daily or less frequent oral dosing.

[0124] Compared to earlier-generation MEK inhibitors, compound 1 has a significantly longer half-life, especially those inhibitors used to treat NF1, which have a half-life of less than 8 hours.

[0125] 1.1.2 Safety and Tolerability In the first two dosing cohorts (n=6), compound 1 was well tolerated with a favorable safety profile, with no drug-related dose interruptions, reductions, or cessations. No drug-related serious adverse events (SAEs) occurred in any dose group and the protocol-defined termination criteria were not met. Importantly, no rash or skin toxicity, gastrointestinal (GI) toxicity, or ocular toxicity has been observed to date at the 2 mg and 4 mg dose levels.

[0126] in conclusion: Unlike first-generation MEK inhibitors for the treatment of NF1 that require twice-daily dosing (BID) and exhibit a short half-life (<8 hours), compound 1 has the potential to achieve prolonged target inhibition due to its long half-life of approximately 70 hours and once-daily dosing (QD). PK profiles showed consistent plasma levels at steady state, as reflected by a low Cmax / Cmin ratio, potentially reducing the risk of Cmax-related toxicities. These findings support the potential benefits of compound 1 in the treatment of NF1, particularly in the treatment of cutaneous and plexiform neurofibromas, cancers, and other MAPK-driven NF1-related conditions.

[0127] The scope of the embodiments provided herein is not limited to the specific embodiments shown in the examples, which are intended to illustrate several aspects of the provided embodiments, and any functionally equivalent embodiments are covered in this disclosure. In fact, various modifications to the embodiments provided herein, other than those shown and described herein, will be apparent to those skilled in the art and are intended to fall within the scope of the appended claims.

[0128] Many references have been cited, and their published content is incorporated into this paper in its entirety through citation.

Claims

1. A pure compound 1, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof: 1。 2. A pure compound 2, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof: 2。 3. A pure compound 3, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof: 3。 4. A pure compound 4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope, or prodrug thereof: 4。 5. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt, tautomer, ester, isotope or prodrug thereof, and a pharmaceutically acceptable excipient or carrier.

6. A pharmaceutical composition comprising the compound of any one of claims 2 to 4 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, ester, isotope or prodrug thereof, and a pharmaceutically acceptable excipient or carrier.

7. A method for treating diseases or conditions with upregulated RAS pathways, such as cancer, RAS protein disorders, and laminopathies, the method comprising administering a therapeutically effective amount of the compound of any one of claims 1 to 4 to a patient suffering from such disease or condition.

8. The method of claim 7, wherein the disease or condition is a neurofibromatosis (NF)-related condition.

Images