Novel RAS inhibitor 3aH-cyclopenta[b]benzofuran-3a-yl

Abstract

The present invention relates to novel compounds and their use as medicines, particularly in the treatment of proliferative diseases. The present invention further relates to pharmaceutical compositions comprising the novel compounds. The present invention also relates to methods for inhibiting cancer cell proliferation or metastasis or inducing cell death of cancer cells in a subject in need thereof. The present invention further relates to methods for inhibiting the proliferation of cell populations sensitive to inhibition of RAS activation, particularly KRAS, HRAS, and NRAS activation, in vitro. The present invention further relates to methods for inhibiting the proliferation of cell populations sensitive to inhibition of the eIF4A complex or binding of the PHB1 / 2 complex in the cell membrane or cytoplasm, in vitro. The present invention also relates to kits comprising formulations containing pharmaceutical compositions comprising compounds according to the present invention.

Description

FIELD OF THE INVENTION

[0001] The present invention relates to novel compounds and their use as medicines, particularly in the treatment of proliferative diseases. The present invention further relates to pharmaceutical compositions comprising said novel compounds. The present invention also relates to methods for inhibiting the proliferation or metastasis of cancer cells or inducing cell death of cancer cells in a subject in need thereof. Furthermore, the present invention relates to methods for inhibiting the proliferation of cell populations sensitive to the inhibition of RAS activation, particularly KRAS, HRAS and NRAS activation, in vitro. Furthermore, the present invention relates to methods for inhibiting the proliferation of cell populations sensitive to the inhibition of the eIF4A complex or the binding of the PHB1 / 2 complex in the cell membrane or cytoplasm, in vitro. Furthermore, the present invention relates to kits comprising formulations containing pharmaceutical compositions comprising compounds according to the present invention. BACKGROUND OF THE INVENTION

[0002] Several approaches are known for the treatment of tumor diseases. The first possibility is the inhibition of the major oncogenes RAS and C-myc, which are activated or upregulated by mutations in the majority of human cancers.

[0003] The RAS oncogene is frequently mutated in human cancers, and among its three isoforms (KRAS, HRAS, and NRAS), KRAS is the most frequently mutated oncogene. As is known, flavagrine-based drugs, such as rocaglamide, a natural antitumor drug and a chemical ligand of prohibitin, inhibit intracellular RAS activation by blocking the interaction between RAS and its effectors at the plasma membrane. Although treatment with rocaglamide inhibits RAS activation in KRAS-mutant cell lines, compounds with better activity against the RAS oncogene remain in demand.

[0004] It is also known that various flavugin derivatives exhibit cytotoxicity. WO 2005 / 113529 A2 describes cyclopenta[b]benzofuran derivatives and their use in the production of medicinal products, in particular for the prevention and / or treatment of acute or chronic diseases.

[0005] WO 2010 / 060891 A1 describes rocaglaol derivatives and the use of these derivatives for preventing or inhibiting the cardiotoxicity of antitumor agents.

[0006] WO 2012 / 0666002 A1 describes flavugin derivatives and their use as neuroprotective, cardioprotective and antitumor agents.

[0007] WO 2017 / 058768 A1 describes compounds that have activity as inhibitors of G12C mutant KRAS protein.

[0008] WO 2020 / 078975 A1 relates to inhibitors of KRAS oncogene activation that are flavugin derivatives, which have the ability to target prohibitin and inhibit KRAS activation.

[0009] N. Ribeiro et al., J. Med. Chem., 2012, 55, 100064 and Thuaud et al., Med. Chem., 2011, 54, 411, relate to flabarin derivatives, particularly FL3 and FL23, which are effective in inhibiting cell proliferation and improving viability at lower doses than rocaglalol.

[0010] Furthermore, it is known that patients frequently develop resistance to RAS oncogene inhibitors, such as KRAS G12 C inhibitors (Tanaka et al., Cancer Discov, 2021, PMID 33824136), and patients treated with KRAS G12 C inhibitors often develop secondary mutations in other RAS isoforms (Awad MM et al., New England Journal of Med., 2021, PMID 34161704).

[0011] EP 2189453 relates to rocaglaol derivatives that can prevent or reduce the cardiotoxicity of antitumor agents.

[0012] Engelmeier D. et al., Journal of Agricultural and Food Chemistry of the American Chemical Society, US, vol. 48, no. 4, 2000-04-01, pp. 1400-1404, relates to a study on plant protection against rice blast fungus. It has been shown that rocaglaol and its derivatives may exhibit antifungal activity.

[0013] Dumontet V. et al., Tetrahedron, Elsevier Science Publishers, Amsterdam, NL, vol. 52, no. 20, 1996-05-13, pp. 6931-6942, relates to a study on the structure of rocaglaol derivatives.

[0014] As mentioned above, several approaches already exist to inhibit the RAS and C-Myc oncogenes, but effectively targeting these genes with small molecules remains a challenge.

[0015] Another option for the treatment of tumor diseases is the inhibition of dysregulated protein translation. In other words, several developments have been made targeting the eukaryotic initiation factor 4A complex (eIF4A complex), which integrates multiple tumor signaling inputs into the translational apparatus, including downstream targets such as C-Myc and cyclin D1, which drive the majority of tumors.

[0016] Eukaryotic initiation factor 4A (eIF4A) is a DEAD-box protein containing an ATPase and an ATP-dependent RNA helicase required for dissolving local secondary structures and allowing ribosomes to access the mRNA template. This factor controls the synthesis of cap-dependent proteins.

[0017] Mammals have three isoforms of eIF4A (eIF4AI, eIF4AII, and eIF4AIII), which share approximately 90% and 65% homology with eIF4AI, the most abundant cellular factor. All isoforms are members of the DEAD-box RNA helicase family, but only the paralogs eIF4AI and eIF4AII are present in the eIF4F complex and are involved in translation initiation.

[0018] WO 2017 / 091585 describes compounds that have activity as eIF4A inhibitors, but the structures of the disclosed compounds differ from those of the compounds of the present invention.

[0019] Prohibitin is an evolutionarily conserved protein, and recent studies have revealed that it plays an important role in RAS activation by enabling RAS-effector interactions at the plasma membrane. Polier et al., Chemistry and Biology, 19, 1093-1104, 2012, showed that rocaglamide targets this interaction (PHB1-CRAF) and inhibits the RAS-CRAF interaction. These effects have been confirmed by several follow-up studies.

[0020] Ernst et al., J. Med. Chem. 2020, 63, 5879, describe that the flavogrine compounds rocaglamide A and zotatifin exhibit inhibitory properties against protein synthesis by stabilizing the untranslatable complex for messenger RNA (mRNA) selection by eIF4A.

[0021] Thus, despite progress in this field, there remains a significant need for compounds that specifically inhibit the activity of RAS and eIF4A, as well as related compositions and methods, particularly with respect to the role of eIF4A in regulating cancer pathways (particularly those driven by c-myc).

[0022] Inhibition of the RAS oncogene likely occurs at the cell membrane via prohibitin-driven nanoclusters (H. Yurugi et al., Journal of Cell Science, 133, 1, 2020), whereas inhibition of eukaryotic initiation factor 4A occurs in the cytoplasm. Thus, two distinct oncoprotein complexes can be targeted to significantly inhibit tumor cell proliferation and survival. Furthermore, activated RAS can also affect the function of the eIF4A complex via one of its effector pathways (MAPK cascades). Furthermore, a recent study revealed that PHB (prohibitin) directly associates with the eukaryotic initiation factor (eIF) 4A complex in the cytoplasm of CLL cells (Largeot et al., Blood, 29 (114) 2023).

[0023] As mentioned above, several approaches already exist to inhibit the RAS oncogene or block the activity of eFI4A, but effectively targeting these genes with small molecules remains a challenge.

[0024] It is therefore an object of the present invention to provide pharmaceutically active compounds capable of inhibiting RAS oncogene activation with high specificity, particularly at nanomolar concentrations in cells. It is a further object of the present invention to provide pharmaceutically active compounds capable of inhibiting RAS and eIF4A activity with high specificity, particularly in cells, at nanomolar concentrations, particularly with regard to the role of both RAS and eIF4A in regulating cancer pathways.

[0025] The above object is achieved by the compounds of formula (I). Summary of the Invention

[0026] The present invention relates to a compound of formula (I) JPEG2026508161000001.jpg91160, a prodrug or isotopically enriched thereof, or a pharmaceutically acceptable salt thereof, wherein: R 1 is CN, R 2is selected from hydrogen and F, R 1 and R 2 Compounds in which both are hydrogen are excluded.

[0027] In particular, the present invention provides compounds of formula (I) JPEG2026508161000002.jpg88165 With respect to a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, the formula: R 1 is CN, R 2 is selected from hydrogen and F.

[0028] Furthermore, the present invention provides compounds of formula (Ia) or (Ib) JPEG2026508161000003.jpg46129 or an enantiomeric mixture comprising a compound of formula (Ia) and (Ib), or a prodrug, isotopically enriched, or pharmaceutically acceptable salt thereof, wherein: R 1 is CN, R 2 is selected from hydrogen and F, R 1 and R 2 Compounds in which both are hydrogen are excluded.

[0029] In particular, the present invention provides compounds of formula (Ia) or (Ib) JPEG2026508161000004.jpg44134 With respect to a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, the formula: R 1 is CN, R 2 is selected from hydrogen and F.

[0030] In particular, the present invention provides compounds of formula (Ia) and (Ib) JPEG2026508161000005.jpg53140, a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, wherein: R 1 is CN, R 2 is selected from hydrogen and F.

[0031] The present invention further relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or in the form of a prodrug or isotopically enriched form thereof, for use as a pharmaceutical.

[0032] The present invention further relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, any of said compounds may be in the form of a pharmaceutically acceptable salt, or a prodrug or isotopically enriched form thereof, for use in the treatment and / or prevention of a proliferative, genetic or inflammatory disease.

[0033] The present invention further relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, any of said compounds being in the form of a pharmaceutically acceptable salt, or a prodrug or isotopically enriched form thereof, for use in the treatment and / or prevention of an inflammatory disease, in particular said inflammatory disease being selected from endometriosis and adenomyosis, particularly when RAS, preferably KRAS, is mutated.

[0034] The present invention further relates to a compound of formula (I), (Ia) or (Ib) as defined above or below, or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, for use in the treatment and / or prevention of a proliferative, genetic or inflammatory disease involving oncogenic RAS or eIF4A complexes that control the expression of genes selected from c-myc and cyclin D1.

[0035] The present invention further relates to compounds of formula (I), (Ia), (Ib), (A), (B), (C), (D), (E), (F), (F), and enantiomeric mixtures thereof as defined above and below, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or a prodrug or isotopically enriched form thereof, for use in the treatment and / or prevention of a genetic disease in which RAS signaling is pathologically involved. In particular, said genetic disease is selected from RASO diseases, in particular CFC (craniofacial skin disease) and NF1 (neurofibromatosis type 1).

[0036] The present invention further relates to compounds of formula (I), (Ia), (Ib), (A), (B), (C), (D), (E), (F), (F), and enantiomeric mixtures thereof as defined above and below, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or a prodrug or isotopically enriched form thereof, for use in the treatment and / or prevention of a genetic disease in which Myc signaling is pathologically involved. In particular, said genetic disease is selected from RASO diseases, in particular CFC (craniofacial skin disease) and NF1 (neurofibromatosis type 1).

[0037] The present invention further relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or a prodrug or isotopically enriched form thereof, for use in the treatment of cancer.

[0038] The present invention further relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or in the form of a prodrug or isotopically enriched form thereof, for use as inhibitors of RAS protein activation.

[0039] The present invention further relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or a prodrug or isotopically enriched form thereof, for use in the treatment or prevention of any disease or condition associated with the activity of a RAS protein (RAS oncogene).

[0040] The present invention further relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, for use in the treatment of proliferative diseases involving RAS signaling, preferably involving KRAS G12V, NRAS G12V, HRAS G12V, KRAS G12C, KRAS G12D, KRAS G12C / Y96D, KRAS G13C, KRAS G13D, KRASG13S, KRAS Q61H, KRAS Q61R, or KRAS Q61K, or involving any activating mutation in KRAS, HRAS, and NRAS, or involving any mutation conferring resistance to RAS inhibitors, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or in the form of a prodrug or isotopically enriched form thereof.

[0041] In particular, the present invention relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or a prodrug or isotopically enriched form thereof, for use in the treatment of proliferative diseases involving RAS signaling.

[0042] In particular, the present invention relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, for use in the treatment of proliferative disorders involving KRAS G12V, NRAS G12V, HRAS G12V, KRAS G12C, KRAS G12D, KRAS G12C / Y96D, KRAS G13C, KRAS G13D, KRASG13S, KRAS Q61H, KRAS Q61R, or KRAS Q61K, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or a prodrug or isotopically enriched form thereof.

[0043] In particular, the present invention relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, for use in the treatment of proliferative diseases involving any activating mutation in KRAS, HRAS and NRAS, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or in the form of a prodrug or isotopically enriched form thereof.

[0044] In particular, the present invention relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or a prodrug or isotopically enriched form thereof, for use in the treatment of proliferative diseases involving any mutation conferring resistance to RAS inhibitors.

[0045] In particular, the present invention relates to a compound of formula (I), (Ia) or (Ib) as defined above and below, an enantiomeric mixture comprising the compounds of formulae (Ia) and (Ib), a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, for use as a ligand of the eIF4A complex, a ligand of prohibitin, or an inhibitor of the eIF4A complex.

[0046] The present invention further relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, for use as inhibitors of eukaryotic initiation factor 4A (eIF4A), wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or in the form of a prodrug or isotopically enriched form thereof.

[0047] In particular, the present invention relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, for use as inhibitors of eukaryotic initiation factor 4A (eIF4A), which controls the expression of cancer driver genes, in particular the expression of c-myc and cyclin D1, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or in the form of a prodrug or isotopically enriched form thereof.

[0048] In particular, the present invention relates to a compound of formula (I), (Ia) or (Ib) as defined above or below, an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, for use in the treatment of a proliferative disease involving eIF4A-Myc signaling.

[0049] The present invention further relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, for use as ligands for prohibitin (PHB1 / 2 complex) in cell membranes or cytoplasm, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or in the form of a prodrug or isotopically enriched form thereof.

[0050] The present invention further relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, for use in the treatment of proliferative diseases involving dysregulation of eIF4A, preferably involving EIF4A1, EIF4A2, EIF4A3 or EIF4F complexes, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or in the form of a prodrug or isotopically enriched form thereof.

[0051] The present invention further relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, for use in the treatment of proliferative disorders involving overexpression of prohibitin (PHB / 2), wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or in the form of a prodrug or isotopically enriched form thereof.

[0052] The present invention further relates to compounds of formula (I), (Ia), (Ib), (A), (B) as defined above and below, and enantiomeric mixtures thereof, for use in the treatment of proliferative diseases in which c-myc is overexpressed and / or cyclin D1 is overexpressed, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or in the form of a prodrug or isotopically enriched form thereof.

[0053] The present invention further relates to pharmaceutical compositions comprising a pharmaceutically effective amount of at least one compound selected from the compounds of formula (I), (Ia), (Ib), (A), (B), enantiomeric mixtures comprising compounds of formula (Ia) and (Ib), or prodrugs, isotopically enriched or pharmaceutically acceptable salts thereof, as defined above and below, and one or more pharmaceutically acceptable carriers.

[0054] The present invention further relates to a pharmaceutical composition comprising a pharmaceutically effective amount of at least one compound selected from the compounds of formula (I), (Ia), (Ib), (A), (B), an enantiomeric mixture comprising the compounds of formula (Ia) and (Ib), or a prodrug or pharmaceutically acceptable salt thereof, for use in the prevention and / or treatment of a proliferative or genetic disease.

[0055] The present invention further relates to a method of inhibiting the growth, proliferation, or metastasis of cancer cells in a subject in need thereof, said method comprising administering to said subject a therapeutically effective amount of at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and below, or an enantiomeric mixture thereof, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt, or in the form of a prodrug or pharmaceutical composition as defined above or below.

[0056] The present invention further relates to a method for inhibiting the proliferation of a cell population sensitive to inhibition of RAS activation, in particular inhibition of KRAS, HRAS or NRAS activation, said method comprising contacting said cell population with at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and below, or an enantiomeric mixture thereof, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt thereof, a prodrug or isotopically enriched form thereof, or in the form of a pharmaceutical composition as defined above and below.

[0057] The present invention further relates to a method for inhibiting the proliferation of a cell population sensitive to the inhibition of eIF4A or its downstream targets, particularly c-myc and / or cyclin D1, in vitro or ex vivo, said method comprising contacting said cell population with at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and below, or an enantiomeric mixture thereof, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt thereof, a prodrug or isotopically enriched form thereof, or a pharmaceutical composition as defined above and below.

[0058] The present invention further comprises: a1) at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and below, or an enantiomeric mixture thereof, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt thereof, or in the form of a prodrug or isotopically enriched form thereof, or a2) A pharmaceutical composition comprising at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and below, or an enantiomeric mixture thereof, any of said compounds being in the form of a pharmaceutically acceptable salt thereof, a prodrug or isotopically enriched form thereof, or a therapeutically acceptable salt thereof, and a pharmaceutically acceptable carrier. A formulation comprising: b) instructions for administering said pharmaceutical composition for the treatment of a disease in which inhibition of RAS activation is effective in treating the disease; The present invention relates to a kit comprising:

[0059] The present invention further comprises: a1) at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and below, or an enantiomeric mixture thereof, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt thereof, or in the form of a prodrug or isotopically enriched form thereof, or a2) a pharmaceutical composition comprising at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and below, or an enantiomeric mixture thereof, wherein any of said compounds may be in the form of a pharmaceutically acceptable salt thereof, a prodrug or isotopically enriched form thereof, or a therapeutically acceptable salt thereof, and a pharmaceutically acceptable carrier; A formulation comprising: b) instructions for administering said pharmaceutical composition for the treatment of a disease in which inhibition of dysregulation of protein translation involving eIF4A is effective in treating the disease; The present invention relates to a kit comprising:

[0060] The present invention further comprises: a1) at least one compound selected from the compounds of formula (I), (Ia), (Ib) as defined above or below, an enantiomeric mixture comprising the compounds of formula (Ia) and (Ib), a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, or a2) a pharmaceutical composition as defined above or below A formulation comprising: b) Instructions for administering said pharmaceutical composition for the treatment of a disease in which inhibition of RAS activation or downstream signaling pathways, in particular C-Myc upregulation and / or cyclin D1 upregulation, is effective in treating the disease. The present invention relates to a kit comprising:

[0061] The present invention has the following advantages: The compounds of the present invention exhibit excellent RAS inhibitory properties, in other words, they qualify as inhibitors of RAS oncogene activation by inhibiting the prohibitin pathway, in particular by inhibiting EGF-induced RAS-GTP loading in cells, as measured by the ability of RAS to bind to effector proteins such as RAF kinase. The compounds of the invention inhibit the activation of RAS because the interaction between RAS and its effectors is disrupted by a defect in the nanoclustering of RAS in the plane of the cell membrane. Potential candidates can be identified by screening compounds of the invention that can inhibit RAS activation by directly inhibiting the interaction of activated RAS (both by EGF and by mutational activation) with the RAS-binding domain (RBD) of CRAF kinase in cells or with other RAS-interacting domains, such as the RA (RAS-associated) domain. - The compound is available on a suitable scale for further development. The compounds are soluble and have favorable pharmacokinetic / pharmacodynamic (PK / PD) properties. They inhibit RAS, including KRAS, NRAS, and HRAS, at nanomolar levels, regardless of whether they are mutated or not. Furthermore, the compound, especially at elevated concentrations and over long periods of time, inhibits the eIF4A complex, which is involved in the translation of several oncogenes, such as c-myc and cyclin D1, and which is the target of so-called STIs (selective translation inhibitors). A dual luciferase assay has been established to measure eIF4A activity in vitro, and the compounds are further screened for inhibition of eIF4A.

[0062] Compounds of formula (I) Unless otherwise stated herein, references in the singular may include the plural, for example, "a" and "an" may refer to one or more.

[0063] The compounds of Formulas (I), (Ia), (Ib), (A), and (B), and their enantiomeric mixtures, can form salts. These salts are also included within the scope of the present invention. As used herein, the term "salt" refers to acidic and / or basic salts formed with inorganic and / or organic acids and bases. Pharmaceutically acceptable salts (i.e., non-toxic, physiologically acceptable salts) are preferred, although other salts are also useful and may be utilized, for example, in preparation, isolation, or purification steps. Salts of the compounds of Formulas (I), (Ia), (Ib), (A), and (B), and their enantiomeric mixtures, can be formed, for example, by reacting the compounds of Formulas (I), (Ia), (Ib), (A), and (B), or their enantiomeric mixtures, with at least one acid or base. The acid or base is added in an amount appropriate for partial or complete neutralization, e.g., an equivalent amount.

[0064] As used herein, unless otherwise indicated, the phrase "pharmaceutically acceptable salt" includes salts containing pharmacologically acceptable anions or cations, such as chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, hydrogen phosphate, dihydrogen phosphate, isonicotinate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, sulfate, benzenesulfonate, p-toluenesulfonate, and palmoate [i.e., 4,4'-methylene-bis-(3-hydroxy-2-naphthoate)] salts.

[0065] Any compound according to the present invention can be prepared and used in the form of a prodrug. Suitable prodrugs have chemically or metabolically cleavable groups that become pharmacologically active compounds in vivo by solvation or under physiological conditions. Prodrugs can be formed by conventional methods by reacting functional groups (such as amino, hydroxy, or carboxy groups) of the compound. Prodrugs often offer advantages such as improved metabolism, potency, solubility, tissue compatibility, or delayed release in mammals.

[0066] The term "prodrug" refers to a pharmaceutically acceptable metabolic precursor of a compound of the present invention. A prodrug may be inactive when administered to a subject in need thereof but is converted in vivo to an active compound of the present invention. Prodrugs are typically rapidly converted in vivo to the parent compound of the present invention, for example, by hydrolysis in blood.

[0067] In other words, the term "prodrug" in the present invention refers to a compound that is metabolized in vivo to become the compound of formula (I) of the present invention. Representative examples of prodrugs are described, for example, in "Practice of Medicinal Chemistry" by C.G. Wermeth (ed.), Academic Press, San Diego, 1996, pp. 671-715, and J. Rautio et al., NATURE REVIEWS, Drug Discovery, Vol. 17, 2018, p. 559. Examples include phosphates, carbonates, carbamates, amino acids, esters (carboxylic acid esters), ethers, amides, peptides, and ureas. In this case, a suitable prodrug may be a compound of formula (I). The hydroxy group in the formula is bonded to any group that is cleaved to form a free hydroxy group when the prodrug of the compound of the present invention is administered to a mammal.

[0068] Myc is a family of regulatory genes and proto-oncogenes that encode transcription factors. The Myc family consists of three related human genes: c-myc (MYC), l-myc (MYCL), and n-myc (MYCN). c-myc (sometimes called MYC) was the first gene in the family discovered due to its homology to the viral gene v-myc. In cancer, c-myc is often constitutively (continuously) expressed. This leads to increased expression of many genes, including those involved in cell proliferation, contributing to the formation of cancer.

[0069] Cyclin D1 is a protein encoded by the CCND1 gene in humans. The CCND1 gene encodes the cyclin D1 protein. The human CCND1 gene is located on the long arm of chromosome 11 (band 11q13). It is 13,388 base pairs long and translates to 295 amino acids. Cyclin D1 is expressed in all tissues of adult humans, except for cells derived from bone marrow stem cell lines (lymphoid and myeloid).

[0070] In the present invention, chemical structures that do not explicitly depict a particular stereochemical orientation generally refer to all possible stereoisomers and mixtures thereof, unless otherwise specified, e.g., JPEG2026508161000006.jpg86146In the formula, * indicates an asymmetric center.

[0071] A "chiral compound" in the sense of the present invention is one that has an inappropriate axis of rotation (S n In the context of the present invention, they are in particular compounds which have at least four asymmetric centers and do not contain S n It is a compound that has no symmetry.

[0072] In the context of this invention, "stereoisomers" are compounds that are identical in structure but differ in the arrangement of their atoms in three-dimensional space.

[0073] "Enantiomers" are stereoisomers that behave as mirror images of each other. For example, the compounds of formula (Ia) and (Ib) are enantiomers. The "enantiomeric excess" (ee) achieved in asymmetric synthesis is given herein by ee [%] = (RS) / (R + S) × 100. R and S are descriptors of the CIP system of two enantiomers and describe the absolute configuration at the asymmetric atom. Enantiomerically pure compounds (ee = 100%) are also called "homochiral compounds."

[0074] "Diastereomers" are stereoisomers that are not mirror images of one another.

[0075] The compounds of the present invention can exist in one or more tautomeric forms, including various isomeric forms and both single tautomers and mixtures of tautomers. The term "isomer" is intended to encompass all isomeric forms of the compounds of the present invention, including tautomeric forms of said compounds.

[0076] Some compounds described herein may have asymmetric centers and therefore exist in different enantiomeric and diastereomeric forms. The compounds of the present invention may be in the form of optical isomers or diastereomers. Accordingly, the present invention encompasses the use and use of the compounds of the present invention described herein in the form of their optical isomers, diastereomers, and mixtures thereof, including racemic mixtures. Optical isomers of the compounds of the present invention can be obtained by known techniques, such as asymmetric synthesis or chiral chromatography, or by chemical separation of stereoisomers using optically active resolving agents.

[0077] Unless otherwise indicated, "stereoisomer" means one stereoisomer of a compound that is substantially free of other stereoisomers of that compound. Thus, a stereoisomerically pure compound having one chiral center will be substantially free of the opposite enantiomer of the compound. A stereoisomerically pure compound having two chiral centers will be substantially free of other diastereomers of the compound. A typical stereoisomerically pure compound contains greater than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of other stereoisomers of the compound, e.g., greater than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of other stereoisomers of the compound, or greater 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 greater than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of other stereoisomers of the compound.

[0078] "Stereoisomer" refers to compounds consisting of the same atoms joined by the same bonds but having different, incompatible three-dimensional structures. The present invention contemplates various stereoisomers and mixtures thereof, and thus includes "enantiomers," which refers to two stereoisomers whose molecules are non-superimposable mirror images of one another.

[0079] The compounds of the present invention, or their pharmaceutically acceptable salts, may contain one or more asymmetric centers and thus may give rise to enantiomers, diastereomers, and other stereoisomeric forms, which, in terms of absolute stereochemistry, may be defined as (R)- or (S)-, or (D)- or (L)-, in the case of amino acids. The present invention is meant to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents or resolved using conventional techniques, such as chromatography and fractional crystallization. Conventional techniques for preparing / isolating individual enantiomers include chiral synthesis from appropriate optically pure precursors, or resolution of the racemate (or racemate of a salt or derivative) using, for example, chiral high-pressure liquid chromatography.

[0080] Relative configuration in stereochemistry (relative stereochemistry) is the arrangement of atoms or groups of atoms described relative to other atoms or groups of atoms in a molecule. In other words, this term describes the position of an atom or group of atoms in space relative to other atoms or groups of atoms located elsewhere in the molecule.

[0081] In stereochemistry (absolute stereochemistry), absolute configuration is the arrangement of atoms or groups of atoms that can be described independently of any other atoms or groups of atoms in a molecule. This type of configuration is defined for chiral molecular entities and their stereochemical description (e.g., R or S).

[0082] Syn refers to the orientation of the substituents (four asymmetric carbon atoms) on the five-membered ring to which they are attached, with all substituents pointing in the same direction relative to the plane of the five-membered ring.

[0083] A racemic mixture or racemic compound is defined as a mixture of compounds consisting of two molecules whose structures are like image and mirror image (= enantiomers) and which are present in the mixture in equimolar amounts, i.e. in a 1:1 (50:50) ratio.

[0084] Furthermore, compounds of formula (I): JPEG2026508161000007.jpg86143 (wherein * indicates an asymmetric center) isomers of formula (Ia), (Ib), (Ic), (Id), (Ie), (If), (Ig), (Ih), (Ii), (Ij), (Ik), (Il), (Im), (In), (Io), and (Ip): JPEG2026508161000008.jpg225132JPEG2026508161000009.jpg225134, where R 1 and R 2 has any of the meanings defined above or below.

[0085] The compounds of formulae (Ia) to (Ip) are characterized by their absolute stereochemistry.

[0086] In a preferred embodiment, the compound of formula (I) is a mixture of at least two enantiomers (Ia) to (Ip), or a mixture of prodrugs or isotopically enriched forms thereof, or a mixture of pharmaceutically acceptable salts thereof, wherein one enantiomer is enriched.

[0087] Preferably, the compound of formula (I) is a mixture of (Ia) and (Ib), or a mixture of pharmaceutically acceptable salts thereof, wherein the enantiomeric excess (ee) of the enantiomer of formula (Ia) is at least 20%, preferably at least 50%, particularly at least 80%, and especially at least 99%.

[0088] Preferably, the compound of formula (I) according to the present invention, a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, wherein R 2 is hydrogen.

[0089] Further preferred are compounds of formula (I) according to the present invention, or prodrugs or pharmaceutically acceptable salts thereof, wherein R 2 is F.

[0090] Another particular embodiment is a compound of formula (I) in which R 1 and R 2 is selected from the definition shown in row 1 of Table 1. Table 1: JPEG2026508161000010.jpg39129

[0091] Another particular embodiment is a compound selected from Compound A, B, and mixtures of each of Compounds A through B with its respective enantiomer. JPEG2026508161000011.jpg56143

[0092] Preferred is a compound of formula (A) or an enantiomeric mixture comprising a compound of formula (A) and its enantiomer, in particular wherein the enantiomer of formula (A) has an enantiomeric excess (ee) of at least 20%, preferably at least 50%, in particular at least 80%, especially at least 99%.

[0093] Preferred is a compound of formula (B) or an enantiomeric mixture comprising a compound of formula (B) and its enantiomer, in particular wherein the enantiomer of formula (B) has an enantiomeric excess (ee) of at least 20%, preferably at least 50%, in particular at least 80%, especially at least 99%.

[0094] Compound A is particularly preferred.

[0095] Compound B is particularly preferred.

[0096] The compounds of the present invention can be synthesized using methods known in the art and known in synthetic organic chemistry, or variations thereof as appreciated by those skilled in the art.

[0097] Pharmaceutical Composition As used herein, the phrase "pharmaceutically acceptable" is used to refer to compounds, materials, compositions, and / or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings or animals without undue toxicity, irritation, allergic response, or other problem or complication, and are commensurate with a reasonable benefit / risk ratio.

[0098] The phrase "therapeutically effective" refers to the amount of each agent that can achieve the goal of ameliorating disease severity and frequency while avoiding the adverse side effects typically associated with alternative therapies. For example, an effective anti-cancer agent may prolong survival or improve quality of life of a patient, inhibit the rapid cell proliferation associated with a neoplasm, or cause regression of the neoplasm.

[0099] As used herein, the terms "treat," "treating," and "treatment" refer to any type of intervention or process performed on a subject, or by administration of an active agent to a subject, to reverse, alleviate, ameliorate, inhibit, or slow or prevent the progression, onset, severity, or recurrence of symptoms, complications, conditions, or biochemical manifestations associated with a disease. In contrast, "prophylaxis" or "prevention" refers to administration to a subject not having a disease to prevent the onset of the disease.

[0100] The term "cell" as used herein is meant to refer to an in vitro, ex vivo, or in vivo cell. In the sense of the present invention, an ex vivo cell may be part of a tissue sample excised from an organism, such as a mammal. In the sense of the present invention, an in vitro cell may be a cell in cell culture. In the sense of the present invention, an in vivo cell is a cell living in an organism, such as a mammal.

[0101] The term "patient" includes humans and animals receiving either therapeutic or prophylactic treatment.

[0102] The term "subject" includes any human or animal. For example, the methods and compositions disclosed herein can be used to treat a subject with cancer.

[0103] (Non-human) animals include all vertebrates, including mammals and non-mammals, for example, cows, sheep, pigs, goats, horses, poultry, dogs, cats, non-human primates, rodents, etc. In one embodiment, the subject is a human subject.

[0104] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid diluent, solvent, excipient, manufacturing aid, etc. (e.g., lubricant), or encapsulating material, that is involved in carrying or transporting a compound of interest from one organ or part of the body to another. Any carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation.

[0105] Suitable other ingredients are the above-mentioned carriers and further additives including adjuvants, preservatives, fillers, flow regulators, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, bittering agents, flavoring agents, antibacterial agents, antifungal agents, lubricants, dispensing agents, etc. Suitable additives are selected depending on the mode of administration and the nature of the dosage form, and are not harmful to the patient.

[0106] The term "pharmaceutical composition" means a composition comprising a compound of the present invention in combination with at least one additional compound selected from the following: a) at least one further pharmaceutically active substance, and b) at least one additional pharmaceutically acceptable carrier and / or excipient.

[0107] RAS The term "RAS inhibitor" refers to an agent that can reduce RAS protein levels, reduce RAS activity levels, and / or inhibit RAS expression levels in cells. RAS inhibitors can be reversible or irreversible inhibitors. As used herein, "RAS" protein refers to a protein that is a member of a family of related proteins expressed in all cell lineages and organs of humans and animals. All members of the RAS protein family belong to a protein class called small GTPases (also known as small G proteins, a family of hydrolases that can bind and hydrolyze GTP) and are involved in transmitting signals within cells (cell signaling). RAS is a typical member of the RAS superfamily of proteins, all of which are related to three-dimensional structures and control diverse cellular behaviors. When RAS is "turned on" by an input signal, it subsequently turns on other proteins, ultimately turning on genes involved in cell growth, differentiation, and survival. Mutations in the RAS gene can lead to the generation of a permanently activated RAS protein, which can cause unintended excessive signaling within cells, even in the absence of an input signal. These signals lead to cell growth and division, so excessive RAS signaling can ultimately lead to cancer. Three RAS genes in humans (HRAS, KRAS, and NRAS) are the most common oncogenes in human cancer. As mentioned above, the most clinically noteworthy genes in the RAS subfamily are HRAS, KRAS, and NRAS. However, the subfamily also includes other members selected from, for example, DIRAS1, DIRAS2, DIRAS3, ERAS, GEM, MRAS, NKIRAS1, NKIRAS2, NRAS, RALA, RALB, RAP1A, RAP1B, RAP2A, RAP2B, RAP2C, RASD1, RASD2, RASL10A, RASL10B, RASL11A, RASL11B, RASL12, REM1, REM2, RERG, RERGL, RRAD, RRAS, and RRAS2.

[0108] At least one compound of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above, and pharmaceutical compositions comprising a compound of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above, may be administered to humans and animals, preferably humans.

[0109] In principle, any mode of administration may be used to deliver the compounds or pharmaceutical compositions of the present invention to a subject. Suitable modes of administration include oral, enteral, parenteral, intravenous, topical, intramuscular, and subcutaneous / transdermal routes.

[0110] The compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above can selectively reduce RAS protein level, reduce RAS activity level, and / or inhibit RAS expression level in cells.For example, the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above can be used to selectively reduce RAS activity level and / or inhibit RAS expression level in cells or in individuals who require reduction of RAS protein level, reduction of RAS activity level, and / or inhibition of RAS expression level by administering an inhibitory amount of the compounds of formula (I), (Ia), (Ib), (A), (B) or one enantiomeric mixture as defined above, or their salts.

[0111] In one embodiment, the present invention provides a combined preparation of a compound of formula (I), (Ia), (Ib), (A), (B) as defined above and one enantiomeric mixture, and / or a pharmaceutically acceptable salt thereof, and / or a prodrug or isotopically enriched form thereof, together with an additional therapeutic agent for simultaneous, separate or sequential use in the treatment and / or prevention of multiple diseases, preferably proliferative diseases (e.g. cancer), in particular diseases associated with the activity of RAS proteins.

[0112] The additional therapeutic agent is selected from a chemotherapeutic agent, a radiotherapeutic agent, an immuno-oncology agent, and combinations thereof.

[0113] In one embodiment, the compound of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above are administered sequentially prior to the administration of the immuno-oncological agent. In another embodiment, the compound of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above are administered simultaneously with the immuno-oncological agent. In yet another embodiment, the compound of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above are administered sequentially after the administration of the immuno-oncological agent.

[0114] In another embodiment, the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above may be formulated with an immuno-oncology agent.

[0115] Immuno-oncology agents include, for example, small molecule drugs, antibodies, or other biological molecules or small molecules. Examples of biological immuno-oncology agents include, but are not limited to, cancer vaccines, antibodies, and cytokines. In one aspect, the antibody is a monoclonal antibody. In another aspect, the monoclonal antibody is a humanized or human antibody.

[0116] In one aspect, the immuno-oncology agent is either (i) a stimulatory (including costimulatory) receptor agonist or (ii) an antagonist of inhibitory (including costimulatory) signals on T cells, both of which result in amplification of antigen-specific T cell responses (often referred to as immune checkpoint modulators).

[0117] Suitable stimulatory and inhibitory molecules are members of the immunoglobulin superfamily (IgSF). One important family of membrane-bound ligands that bind to costimulatory or co-inhibitory receptors is the B7 family, which includes B7-1, B7-2, B7-H1 (PD-L1), B7-DC (PD-L2), B7-H2 (ICOS-L), B7-H3, B7-H4, B7-H5 (VISTA), and B7-H6. Another family of membrane-bound ligands that bind to costimulatory or coinhibitory receptors is CD40 and CD40L, OX-40, OX-40L, CD70, CD27L, CD30, CD30L, 4-1BBL, CD137 (4-1BB), TRAIL / Apo2-L, TRAILR1 / DR4, TRAILR2 / DR5, TRAILR3, TRAILR4, OPG, RANK, RANKL, TWEAKR / Fnl4, TWEAK, BAFFR, EDAR, XEDAR, TACI, APRIL, BCMA, LTpR, LIGHT, DcR3, HVEM, VEGETL1A, TRAMP / DR3, EDAR, EDA1, XEDAR, EDA2, TNFR1, lymphotoxin a / TNFp, TNFR2, TNFa, LTpR, and lymphotoxin a. The TNF family of molecules binds to cognate TNF receptor family members, including 1b2, FAS, FASL, RELT, DR6, TROY, and NGFR.

[0118] In one aspect, T cell responses can be stimulated by a combination of compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above with one or more of the following: (i) antagonists of proteins that inhibit T cell activation (e.g., immune checkpoint inhibitors), such as CTLA-4, PD-1, PD-L1, PD-L2, LAG-3, TIM-3, galectin-9, CEACAM-1, BTLA, CD69, galectin-1, TIGIT, CD113, GPR56, VISTA, 2B4, CD48, GARP, PD1H, LAIR1, TIM-1, and TIM-4, and (ii) Stimulators of proteins that stimulate T cell activation, such as B7-1, B7-2, CD28, 4-1BB (CD137), 4-1BBL, ICOS, ICOS-L, OX40, OX40L, GITR, GITRL, CD70, CD27, CD40, DR3, and CD28H.

[0119] Other drugs that can be used in combination with the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above for the treatment of cancer include antagonists of inhibitory receptors on NK cells or stimulators of activating receptors on NK cells. For example, the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above can be used in combination with KIR antagonists such as lirilumab.

[0120] Other agents for combination therapy include agents that inhibit or deplete macrophages or monocytes, including, but not limited to, CSF-1R antagonists such as CSF-1R antagonist antibodies, including RG7155.

[0121] Combination therapy is intended to encompass the administration of the therapeutic agents sequentially, i.e., administering each therapeutic agent at different times, as well as the administration of the therapeutic agent or at least two therapeutic agents substantially simultaneously.

[0122] Substantially simultaneous administration can be achieved, for example, by administering to a subject a single dosage form containing a fixed ratio of each therapeutic agent, or multiple single dosage forms of each therapeutic agent. Sequential or substantially simultaneous administration of each therapeutic agent can be achieved by any appropriate route of administration, including, but not limited to, oral administration, intravenous administration, intramuscular administration, and direct absorption through mucosal tissue. The therapeutic agents can be administered by the same route or by different routes. For example, the first therapeutic agent of a selected combination may be administered intravenously, while the other therapeutic agents of the combination may be administered orally. Alternatively, for example, all therapeutic agents may be administered orally, or all therapeutic agents may be administered intravenously. Combination therapy also includes further combination of the above-mentioned therapeutic agents with other biologically active ingredients and non-drug treatments (e.g., surgery or radiation therapy). When combination therapy further includes a non-drug treatment, the non-drug treatment may be administered at an appropriate time, as long as beneficial effects are obtained from the synergistic action of the combined therapeutic agent and non-drug treatment. For example, in appropriate cases, the non-drug treatment can be temporarily separated from the administration of the therapeutic agent, perhaps by days or weeks, and the beneficial effects still be achieved.

[0123] The types of cancer that may be treated with the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above include, but are not limited to, cancer of the prostate, colon, rectum, pancreas, cervix, stomach, endometrium, brain, liver, bladder, ovary, testis, head, neck, skin (including melanoma and basal cell carcinoma), mesothelial lining, white blood cells (including lymphoma and leukemia), esophagus, breast, muscle, connective tissue, lung (including small cell lung carcinoma and non-small cell carcinoma), adrenal gland, thyroid, kidney, or bone, or glioblastoma, mesothelioma, renal cell carcinoma, gastric cancer, sarcoma (including Kaposi's sarcoma), choriocarcinoma, basal cell carcinoma of the skin, hematological malignancies (including blood, bone marrow, and lymph nodes), or testicular seminoma.

[0124] To treat a RAS protein-associated disease, disorder, or condition, one or more additional pharmaceutical agents or therapies, for example, antiviral agents, chemotherapeutic or other anti-cancer agents, immunostimulants, immunosuppressants, radiation therapy, anti-tumor and anti-viral vaccines, cytokine therapy (e.g., IL2 and GM-CSF), and / or tyrosine kinase inhibitors, can optionally be used in combination with the compounds of Formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above. These agents can be combined with the compound in a single dosage form, or they can be administered simultaneously or sequentially in separate dosage forms.

[0125] Suitable chemotherapeutic or other anti-cancer agents include, for example, alkylating agents (including, but not limited to, nitrogen mustards, ethyleneimine derivatives, alkylsulfonates, nitrosoureas, and triazenes) such as uracil mustard, chlormethine, cyclophosphamide (CYTOXAN®), ifosfamide, melphalan, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, and temozolomide.

[0126] Agents suitable for use in combination with the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above in the treatment of melanoma include dacarbazine (DTIC), optionally in combination with other chemotherapeutic agents such as carmustine (BCNU) and cisplatin, the "Dartmouth therapy" consisting of DTIC, BCNU, cisplatin, and tamoxifen, and the combination of cisplatin, vinblastine, DTIC, temozolomide, or YERVOY™. The compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above may also be used in combination with immunotherapeutic agents, including cytokines such as interferon alpha, interleukin 2, and tumor necrosis factor (TNF), in the treatment of melanoma. The compounds of formula (I), (Ia), (Ib), (A), and (B) as defined above and a mixture of one enantiomer may be used in combination with vaccine therapy in the treatment of melanoma. Anti-melanoma vaccines are similar in some ways to antiviral vaccines used to prevent diseases caused by viruses such as polio, measles, and mumps. Weakened melanoma cells or parts of melanoma cells, called antigens, may be injected into patients to stimulate the body's immune system to destroy the melanoma cells.

[0127] Melanoma limited to the arms or legs may be treated using a combination of drugs, including one or more compounds of Formula (I), (Ia), (Ib), (A), and (B) as defined above, and one enantiomeric mixture, using hyperthermic isolated limb perfusion. In this treatment protocol, the blood circulation of the affected limb is temporarily isolated from the rest of the body, and high doses of chemotherapy drugs are infused into the arteries supplying that limb. This allows high doses to be delivered to the tumor area without exposing internal organs to these drugs, which can cause serious side effects. The fluid is usually warmed to 38.9°C to 40°C. Melphalan is the drug most commonly used in this chemotherapy. It can be administered with another drug called a tumor necrosis factor (TNF) inhibitor.

[0128] Suitable chemotherapeutic or other anti-cancer agents include, for example, antimetabolites (including, but not limited to, antifolates, pyrimidine analogs, purine analogs, and adenosine deaminase inhibitors) such as methotrexate, 5-fluorouracil, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, pentostatin, and gemcitabine.

[0129] Suitable chemotherapeutic or other anti-cancer agents further include certain natural products and their derivatives (e.g., vinca alkaloids, antitumor antibiotics, enzymes, lymphokines, and epipodophyllotoxins), such as, for example, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, ara-C, paclitaxel (taxol), mithramycin, deoxyco-formycin, mitomycin-C, L-asparaginase, interferons (especially IFN-α), etoposide, and teniposide.

[0130] Other cytotoxic agents include navelbene, CPT-11, anastrazole, letrazole, capecitabine, reloxafine, and droloxafine.

[0131] Also suitable are cytotoxic agents such as epidophyllotoxins, antitumor enzymes, topoisomerase inhibitors, procarbazine, mitoxantrone, platinum complexes such as cisplatin and carboplatin, biological response modifiers, growth inhibitors, antihormonal therapies, leucovorin, tegafur, and hematopoietic growth factors.

[0132] Other anti-cancer agents include antibody therapeutics such as trastuzumab (Herceptin®), antibodies against costimulatory molecules such as CTLA-4, 4-1BB and PD-1, or antibodies against cytokines (IL-1O or TGF-b).

[0133] Other anti-cancer drugs include those that block immune cell migration, such as antagonists to chemokine receptors including CCR2 and CCR4.

[0134] Other anti-cancer drugs include those that boost the immune system, such as adjuvants or adoptive T-cell transfer.

[0135] Anti-cancer vaccines include dendritic cells, synthetic peptides, DNA vaccines, and recombinant viruses.

[0136] In a specific embodiment of the present invention, at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and one enantiomeric mixture, and at least one chemotherapeutic agent are administered to a patient simultaneously or sequentially. In other words, at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and one enantiomeric mixture may be administered first, or at least one chemotherapeutic agent may be administered first, or at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and one enantiomeric mixture may be administered simultaneously. In addition, when two or more compounds of formula (I), (Ia), (Ib), (A), (B) as defined above, one enantiomeric mixture, and / or chemotherapeutic agents are used, these compounds may be administered in any order.

[0137] The present invention also provides pharmaceutical compositions comprising a therapeutically effective amount of one or more compounds of formula (I), (Ia), (Ib), (A), (B) as defined above, and one enantiomeric mixture, formulated together with one or more pharmaceutically acceptable carriers (excipients) and / or diluents, and optionally one or more additional therapeutic agents as described above.

[0138] The compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above may be administered by any suitable route, preferably in the form of a pharmaceutical composition adapted for such a route, in a dose effective for the intended treatment. The compounds of formula (I), (Ia), (Ib), (A), (B) and compositions thereof, as well as enantiomeric mixtures, as defined above, can be administered for any of the uses described herein by any suitable means, for example, oral administration such as tablets, capsules (including sustained-release or time-release formulations), pills, powders, granules, elixirs, tinctures, suspensions (including nanosuspensions, microsuspensions, and spray-dried dispersions), syrups, and emulsions; sublingual administration; buccal administration; parenteral administration such as subcutaneous, intravenous, intramuscular, or intrasternal injection, or infusion (e.g., sterile injectable aqueous or non-aqueous solutions or suspensions); nasal administration, including administration to the nasal mucosa, such as by inhalation spray; and topical administration, such as rectal administration in the form of a cream or ointment, or in the form of a suppository. They can be administered alone, but will usually be administered with a pharmaceutical carrier selected based on the chosen route of administration and standard pharmaceutical practice.

[0139] For oral administration, the pharmaceutical composition may be in the form of, for example, a tablet, capsule, liquid capsule, suspension, or liquid. The pharmaceutical composition is preferably prepared in the form of a dosage unit containing a specific amount of the active ingredient. For example, the pharmaceutical composition may be provided as a tablet or capsule containing about 0.1 to 1000 mg, preferably about 0.25 to 250 mg, and more preferably about 0.5 to 100 mg of the active ingredient. The appropriate daily dosage for humans or animals can vary greatly depending on the patient's condition and other factors, but can be determined using routine methods.

[0140] The pharmaceutical compositions contemplated herein can be orally administered, for example, via any acceptable and suitable oral formulation. Exemplary oral formulations include, but are not limited to, tablets, troches, lozenges, aqueous and oily suspensions, dispersible powders or granules, emulsions, hard and soft capsules, liquid capsules, syrups, and elixirs. Pharmaceutical compositions for oral administration can be prepared according to any method known in the art for preparing pharmaceutical compositions for oral administration. To provide a pharmaceutically palatable formulation, the pharmaceutical composition according to the present invention may contain at least one agent selected from sweeteners, flavoring agents, bittering agents, coloring agents, demulcents, antioxidants, and preservatives.

[0141] Tablets can be prepared, for example, by mixing at least one compound of formula (I), (Ia), (Ib), (A), or (B) as defined above and one enantiomeric mixture thereof, and / or at least one pharmaceutically acceptable salt thereof, with at least one non-toxic pharmaceutically acceptable excipient suitable for the manufacture of tablets. Exemplary excipients include, but are not limited to, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, sodium phosphate, etc.; granulating and disintegrating agents such as microcrystalline cellulose, croscarmellose sodium, corn starch, and alginic acid; binders such as starch, gelatin, polyvinylpyrrolidone, and acacia; and lubricants such as magnesium stearate, stearic acid, and talc. Tablets may also be uncoated or coated by known techniques to mask the unpleasant taste of unpleasant-tasting drugs or to delay disintegration and absorption of the active ingredient in the gastrointestinal tract, thereby prolonging the effect of the active ingredient. Examples of water soluble taste masking agents include, but are not limited to, hydroxypropyl methylcellulose and hydroxypropyl cellulose. Examples of time delay agents include, but are not limited to, ethyl cellulose and cellulose acetate butyrate.

[0142] Hard gelatin capsules can be prepared, for example, by mixing at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and one enantiomeric mixture, and / or at least one salt thereof, with at least one inert solid diluent, such as, for example, calcium carbonate, calcium phosphate, and kaolin.

[0143] Soft gelatin capsules can be prepared, for example, by mixing at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and one enantiomeric mixture, and / or at least one pharmaceutically acceptable salt thereof, with at least one water-soluble carrier, such as, for example, polyethylene glycol, and at least one oily medium, such as, for example, peanut oil, liquid paraffin, and olive oil.

[0144] Aqueous suspensions can be prepared, for example, by mixing at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and one enantiomeric mixture, and / or at least one pharmaceutically acceptable salt thereof, and / or a prodrug or isotopically enriched form thereof, with at least one excipient suitable for the manufacture of aqueous suspensions. Exemplary excipients suitable for the manufacture of aqueous suspensions include, but are not limited to, suspending agents, such as sodium carboxymethylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, sodium alginate, alginic acid, polyvinylpyrrolidone, gum tragacanth, and gum acacia; dispersing or wetting agents, such as natural phospholipids such as lecithin; condensation products of alkylene oxides with fatty acids, such as polyoxyethylene stearate; condensation products of ethylene oxide with long chain aliphatic alcohols, such as heptadecaethyleneoxycetanol; condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols, such as polyoxyethylene sorbitol monooleate; and condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as polyethylene sorbitan monooleate. The aqueous suspensions may also contain at least one preservative (e.g., ethyl parahydroxybenzoate and n-propyl parahydroxybenzoate); at least one coloring agent; at least one flavoring agent; and / or at least one sweetening agent (including, but not limited to, sucrose, saccharin, and aspartame).

[0145] Oily suspensions can be prepared, for example, by suspending at least one compound of formula (I), (Ia), (Ib), (A), or (B) as defined above, and one enantiomeric mixture, and / or at least one pharmaceutically acceptable salt thereof, and / or a prodrug or isotopically enriched form thereof, in a vegetable oil such as peanut oil, olive oil, sesame oil, or coconut oil, or in a mineral oil such as liquid paraffin. Oily suspensions can also contain at least one thickening agent, such as beeswax, hard paraffin, and cetyl alcohol. To provide a palatable oily suspension, at least one sweetener and / or at least one flavoring agent, as already described above, can be added to the oily suspension. Oily suspensions can further contain at least one preservative, including, but not limited to, antioxidants such as butylated hydroxyanisole and α-tocopherol.

[0146] Dispersible powders and granules can be prepared, for example, by mixing at least one compound of formula (I), (Ia), (Ib), (A), or (B) as defined above, and one enantiomeric mixture thereof, and / or at least one pharmaceutically acceptable salt thereof, and / or a prodrug or isotopically enriched form thereof, with at least one dispersing agent and / or wetting agent, at least one suspending agent, and / or at least one preservative. Suitable dispersing agents, wetting agents, and suspending agents have already been described above. Examples of preservatives include, but are not limited to, antioxidants such as ascorbic acid. Dispersible powders and granules can also contain at least one excipient, including, but not limited to, sweeteners, flavoring agents, and coloring agents.

[0147] Emulsions of at least one compound of formula (I), (Ia), (Ib), (A), or (B) as defined above and one enantiomeric mixture thereof, and / or at least one pharmaceutically acceptable salt thereof, and / or a prodrug or isotopically enriched version thereof, can be prepared, for example, as an oil-in-water emulsion. The oil phase of the emulsion containing the compound of formula (I), (Ia), (Ib), (A), or (B) and the enantiomeric mixture as defined above can be composed of known ingredients in a known manner. This oil phase can be provided by, but is not limited to, vegetable oils such as olive oil and peanut oil, mineral oils such as liquid paraffin, and mixtures thereof. This phase can contain only an emulsifier, or it can contain a mixture of at least one emulsifier with a fat or oil, or both a fat and an oil. Suitable emulsifiers include, but are not limited to, naturally occurring phospholipids (e.g., soybean lecithin), esters or partial esters derived from fatty acids and hexitol anhydrides (e.g., sorbitan monooleate), and condensation products of partial esters with ethylene oxide (e.g., polyoxyethylene sorbitan monooleate).

[0148] Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier, which acts as a stabilizer. It is also preferred to include both an oil and a fat. The emulsifier, with or without a stabilizer, constitutes a so-called emulsifying wax, which, together with the oil and fat, constitutes a so-called emulsifying ointment base, forming the oily dispersed phase of the cream formulation. The emulsifier may also contain sweeteners, flavorings, preservatives, and / or antioxidants. Emulsifiers and emulsion stabilizers used in the formulations of the present invention include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate, sodium lauryl sulfate, glyceryl distearate (alone or in combination with a wax), or other substances known in the art.

[0149] The compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture thereof, and / or at least one pharmaceutically acceptable salt thereof, and / or prodrug or isotopically enriched thereof, as defined above, can also be delivered, for example, intravenously, subcutaneously, and / or intramuscularly, via any pharmaceutically acceptable and suitable injectable form. Examples of injectable forms include, but are not limited to, sterile aqueous solutions containing acceptable vehicles and solvents, such as water, Ringer's solution, and isotonic sodium chloride solution; sterile oil-in-water microemulsions, and aqueous or oily suspensions.

[0150] Formulations for parenteral administration may be in the form of isotonic sterile injection solutions or suspensions, such as aqueous or non-aqueous solutions. These solutions and suspensions can be prepared from sterile powders or granules using one or more of the carriers or diluents mentioned for oral formulations, or other suitable dispersing, wetting, or suspending agents. The compounds can be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, tragacanth gum, and / or various buffers. Other adjuvants and administration methods are well known and widely understood in the pharmaceutical arts. The active ingredient may be administered by injection in a composition with a suitable carrier, including saline, dextrose, and water, or in a composition with a cyclodextrin solubilizer (i.e., Captisol), a cosolvent solubilizer (i.e., propylene glycol), or a micelle solubilizer (i.e., Tween 80).

[0151] Sterile injectable preparations may also be sterile injectable solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, for example, as solutions in 1,3-butanediol. Among the acceptable vehicles and solvents that may be used are water, Ringer's solution, and isotonic sodium chloride solution. Sterile, solid oils are also conventionally used as solvents or suspending media. For this purpose, any bland, solid oil can be used, including synthetic mono- or diglycerides. Fatty acids, such as oleic acid, are also used in the preparation of injectables.

[0152] Sterile injectable oil-in-water microemulsions can be prepared, for example, by: 1) dissolving at least one compound of formula (I), (Ia), (Ib), (A), (B), and one enantiomeric mixture as defined above in an oil phase, such as a mixture of soybean oil and lecithin; 2) combining the compound of formula (I), (Ia), (Ib), (A), (B), and one enantiomeric mixture as defined above, comprising the oil phase, with a mixture of water and glycerol; and 3) treating the combination to form a microemulsion.

[0153] Sterile aqueous or oily suspensions can be prepared according to methods known in the art. For example, sterile aqueous solutions or suspensions can be prepared using non-toxic parenterally acceptable diluents or solvents, such as 1,3-butanediol, and sterile oily suspensions can be prepared using sterile, non-toxic acceptable solvents or suspending media, such as sterile solid oils, such as synthetic monoglycerides or diglycerides, and fatty acids, such as oleic acid.

[0154] Pharmaceutically acceptable carriers are formulated according to many factors within the understanding of those skilled in the art. These factors include, but are not limited to, the type and nature of the active agent being formulated, the subject to whom the drug containing the composition will be administered, the intended route of administration of the composition, and the targeted therapeutic indication. Pharmaceutically acceptable carriers include both aqueous and non-aqueous liquid media, as well as various solid and semi-solid dosage forms. Such carriers may contain many different components and additives in addition to the active agent. Such additional components are included in the formulation for various reasons well known to those skilled in the art, such as stabilization of the active agent or as a binder. Suitable pharmaceutically acceptable carriers and factors related to their selection are described in various available sources, such as Allen, LV Jr. et al., Remington: The Science and Practice of Pharmacy (Vol. 2), 22nd Edition (2012), Pharmaceutical Press.

[0155] Pharmaceutically acceptable carriers, adjuvants, and vehicles that can be used in the pharmaceutical compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS), such as d-α-tocopherol polyethylene glycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tween, polyethoxylated castor oil such as CREMOPHOR surfactants (BASF), or other similar polymeric delivery matrices, serum proteins, such as human serum albumin, buffer substances, such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol, and wool fat. Cyclodextrins, such as α-, β-, γ-cyclodextrin, or chemically modified derivatives, such as hydroxyalkyl cyclodextrins (including 2- and 3-hydroxypropyl-cyclodextrin), or other solubilizing derivatives, may also be advantageously used to enhance delivery of compounds of the formulae described herein.

[0156] The pharmaceutically active compounds of the present invention can be processed according to conventional pharmaceutical methods to produce medicaments for administration to patients, including humans and other mammals. The pharmaceutical compositions may be subjected to conventional pharmaceutical procedures such as sterilization and / or contain conventional adjuvants such as preservatives, stabilizers, wetting agents, emulsifiers, buffers, etc. Tablets and pills can also be prepared with enteric coatings. Such compositions may also contain adjuvants such as wetting agents, sweeteners, flavoring agents, and fragrances.

[0157] For therapeutic purposes, the active compound of the present invention is usually combined with one or more adjuvants suitable for the indicated administration route.For oral administration, the compound can be mixed with lactose, sucrose, starch powder, cellulose esters of alkanoic acid, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphate and sulfate, gelatin, gum arabic, sodium alginate, polyvinylpyrrolidone, and / or polyvinyl alcohol, and then tableted or encapsulated for easy administration.Such capsules or tablets can also contain controlled-release formulations, which can be provided in a dispersion of the active compound in hydroxypropylmethylcellulose.

[0158] The amount of compound administered and the dosage regimen for treating a disease state using the compounds and / or compositions of the present invention depend on various factors, including the age, weight, sex, and health of the subject, the type of disease, the severity of the disease, the route and frequency of administration, and the particular compound used. Thus, dosage regimens can vary widely but can be routinely determined using standard methods. A daily dosage of about 0.001 to 100 mg / kg body weight, preferably about 0.0025 to about 50 mg / kg body weight, and most preferably about 0.005 to 10 mg / kg body weight, may be appropriate. The daily dosage may be administered one to four times daily. Other dosage schedules include once-weekly administration and once-every-two-day administration.

[0159] The pharmaceutical compositions of the present invention comprise at least one compound of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above, and / or at least one pharmaceutically acceptable salt thereof and / or a prodrug or isotopically enriched form thereof, and optionally an additional agent selected from any pharmaceutically acceptable carrier, adjuvant, and vehicle. Alternative compositions of the present invention comprise a compound of formula (I), (Ia), (Ib), (A), (B) and an enantiomeric mixture as defined above, or a prodrug or isotopically enriched form thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

[0160] The present invention also includes pharmaceutical kits useful for treating or preventing, for example, RAS protein-related diseases. Accordingly, the present invention also relates to a kit comprising: a) a formulation comprising a pharmaceutical composition comprising one of the compounds of formula (I), (Ia), (Ib), (A), (B) and one of the enantiomeric mixtures defined above, or a therapeutically acceptable salt and / or prodrug or isotopically enriched form thereof, and a pharmaceutically acceptable carrier; and b) instructions for administering the pharmaceutical composition to treat a disease in which inhibition of RAS activation is effective in treating the disease.

[0161] Such kits may optionally further include one or more of a variety of conventional pharmaceutical kit components, such as containers containing one or more pharmaceutically acceptable carriers, additional containers as would be readily apparent to one of skill in the art, etc. Instructions indicating the amounts of components to be administered, administration guidelines, and / or guidelines for mixing the components may also be included in the kit, either as an insert or label.

[0162] It will be appreciated that the administration regimen for the compounds of the present invention will vary depending on known factors such as the pharmacodynamic properties of the particular drug, the method and route of administration, the patient's race, age, sex, health, condition and weight, the nature and extent of symptoms, type of concomitant treatment, frequency of treatment, route of administration, the patient's renal and hepatic function, and the desired effect.

[0163] As a general guideline, the daily oral dose of each active ingredient, when used to achieve the desired effect, will range from about 0.001 to about 5000 mg / day, preferably from about 0.01 to about 1000 mg / day, and most preferably from about 0.1 to about 250 mg / day. For intravenous administration, the most preferred dose will be in the range of about 0.01 to about 10 mg / kg / minute during a constant rate infusion. The compounds of Formula (I), (Ia), (Ib), (A), (B), and one enantiomeric mixture may be administered once daily, or the total daily dose may be divided into two, three, or four doses.

[0164] The compounds are typically administered in admixture with suitable pharmaceutical diluents, excipients, or carriers (collectively referred to herein as pharmaceutical carriers) appropriately selected according to the intended administration form, such as oral tablets, capsules, elixirs, and syrups, and in accordance with conventional pharmaceutical practice. Dosage forms (pharmaceutical compositions) suitable for administration may contain from about 1 mg to about 200 mg of active ingredient per dosage unit. In these pharmaceutical compositions, the active ingredient is usually present in an amount of from about 0.1 to 95% by weight, based on the total weight of the composition.

[0165] A typical capsule for oral administration contains at least one compound of Formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture (250 mg), lactose (75 mg), and magnesium stearate (15 mg), which mixture is passed through a 60-mesh sieve and filled into a No. 1 gelatin capsule.

[0166] A typical injection is prepared by aseptically placing at least one compound of Formula (I), (Ia), (Ib), (A), or (B) and one enantiomeric mixture (250 mg) into a vial, aseptically lyophilizing the vial, and sealing the vial. At the time of use, the contents of the vial are mixed with 2 mL of saline to prepare the injection.

[0167] The present invention includes within its scope pharmaceutical compositions comprising, as active ingredient, a therapeutically effective amount of at least one compound of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above, alone or in combination with a pharmaceutical carrier. Optionally, the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above can be used alone or in combination with other compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above, or in combination with one or more other therapeutic agents, such as anti-cancer agents or other pharmaceutically active substances.

[0168] Regardless of the route of administration selected, the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above, which may be used in a suitable hydrated form, and / or the pharmaceutical compositions of the present invention, are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those skilled in the art.

[0169] Actual dosage levels of the active ingredients in the pharmaceutical compositions of the present invention may be varied so as to provide an amount of the active ingredient effective to achieve the desired therapeutic response for a particular patient, composition, and method of administration without causing toxicity to the patient.

[0170] The selected dosage level will depend upon a variety of factors, including the activity of the particular compound of formula (I), (Ia), (Ib), (A), (B) as defined above, or one enantiomeric mixture thereof, used, or an ester, salt, or amide thereof, the route of administration, the time of administration, the rate of excretion or metabolism of the particular compound used, the rate and extent of absorption, the duration of treatment, other drugs, compounds, and / or substances used in combination with the particular compound used, the age, sex, weight, condition, general health, and medical history of the patient being treated, and similar factors well known in the medical field.

[0171] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian can start the dosage of the compound of Formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture used in the pharmaceutical composition as defined above at a lower amount than required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved.

[0172] Generally, a suitable daily dose of the compounds of formula (I), (Ia), (Ib), (A), and (B) and one enantiomeric mixture thereof as defined above will be the minimum dose effective to produce a therapeutic effect. Such an effective dose will generally depend on the factors described above. Generally, oral, intravenous, intracerebroventricular, and subcutaneous doses of the compounds of formula (I), (Ia), (Ib), (A), and (B) and one enantiomeric mixture thereof as defined above to a patient will range from about 0.01 to about 50 mg per kg of body weight per day.

[0173] If desired, the effective daily dose of the active compound may be administered as two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally in unit dosage forms. In certain embodiments of the invention, administration is once daily.

[0174] While it is possible for the compounds of formula (I), (Ia), (Ib), (A), (B) and single enantiomeric mixtures as defined above to be administered alone, it is preferred to administer said compounds as pharmaceutical formulations (compositions).

[0175] The above-mentioned other therapeutic agents, when used in combination with the compounds of Formula (I), (Ia), (Ib), (A), (B) and single enantiomeric mixtures defined above, may be used, for example, in amounts indicated in the Physicians' Desk Reference (PDR) or in amounts determined by one skilled in the art. In the methods of the present invention, such other therapeutic agents may be administered before, simultaneously with, or after the administration of the compounds of the present invention.

[0176] Eukaryotic initiation factor 4A Given the complexity of the mechanisms governing the basic survival of eukaryotic cells, it is not surprising that regulation occurs at multiple stages of protein synthesis. Human translational regulation has become a focus of increasing research interest due to its association with various diseases. Homologous genes for many factors involved in human translation are shared among various eukaryotes. Protein synthesis from mature messenger RNA in eukaryotes is divided into the steps of translation initiation, elongation, and termination, with translation initiation being the rate-limiting step. The bottleneck in the translation initiation process occurs just before the ribosome binds to 5'm7GTP, which is facilitated by multiple proteins. It is at this step that limitations due to stress and amino acid deprivation come into play.

[0177] Eukaryotic initiation factor complex 2 (eIF2) forms a ternary complex with GTP and the initiation factor Met-tRNA. This process is regulated by guanine nucleotide exchange and phosphorylation and serves as a key regulator of gene expression bottlenecks. Before translation can proceed to the elongation stage, several initiation factors must facilitate synergistic interaction between the ribosome and mRNA, ensuring that the mRNA's 5'UTR is sufficiently free of secondary structure. Such binding is facilitated by group 4 eukaryotic initiation factors. eIF4F is involved not only in normal translational control but also in cancer cell transformation and progression.

[0178] eIF4F is responsible for binding capped mRNA to the 40S ribosomal subunit via eIF3. The mRNA cap is bound by eIF4E (25 kDa), and eIF4G (185 kDa) serves as the scaffold for the complex. Meanwhile, eIF4A (46 kDa), an ATP-dependent RNA helicase, processes the secondary structure of the mRNA 5'UTR, facilitating ribosome binding and subsequent translation. These three proteins are collectively referred to as eIF4F. eIF4A also requires eIF4B (80 kDa) for maximal activity, and eIF4B is activated by eIF4H (25 kDa). Upon binding to the 5' cap of the mRNA, this 48S complex (usually) searches for an AUG start codon and initiates translation.

[0179] At least one compound of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above, and pharmaceutical compositions comprising a compound of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above, may be administered to humans and animals, preferably humans.

[0180] In principle, any mode of administration may be used to deliver the compounds or pharmaceutical compositions of the present invention to a subject. Suitable modes of administration include oral, enteral, parenteral, intravenous, topical, intramuscular, and subcutaneous routes.

[0181] The compounds of formula (I), (Ia), (Ib), (A), and (B) and one enantiomeric mixture thereof as defined above can selectively reduce the activity of eIF4A, reduce the activity level of eIF4A, and / or inhibit the activity of eIF4 in the cytoplasm. For example, the compounds of formula (I), (Ia), (Ib), (A), and (B) and one enantiomeric mixture thereof as defined above can be used to selectively reduce the activity level of eIF4A and / or inhibit eIF4A in the cytoplasm or in an individual in need of reduced eIF4A activity, reduced eIF4A activity, and / or inhibited eIF4A activity level by administering an inhibitory amount of the compounds of formula (I), (Ia), (Ib), (A), and (B) or one enantiomeric mixture thereof as defined above, or a salt thereof.

[0182] In one embodiment, the present invention provides a combined preparation of a compound of formula (I), (Ia), (Ib), (A), (B) as defined above and one enantiomeric mixture, and / or a pharmaceutically acceptable salt thereof, and / or a prodrug or isotopically enriched form thereof, together with an additional therapeutic agent for simultaneous, separate or sequential use in the treatment and / or prevention of multiple diseases, preferably proliferative diseases (e.g. cancer), in particular diseases associated with the activity of eIF4A.

[0183] The additional therapeutic agent is selected from a chemotherapeutic agent, a radiotherapeutic agent, an immuno-oncology agent, and combinations thereof.

[0184] In one embodiment, the compound of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above are administered sequentially prior to the administration of the immuno-oncological agent. In another embodiment, the compound of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above are administered simultaneously with the immuno-oncological agent. In yet another embodiment, the compound of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above are administered sequentially after the administration of the immuno-oncological agent.

[0185] In another embodiment, the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above may be formulated with an immuno-oncology agent.

[0186] Immuno-oncology agents include, for example, small molecule drugs, antibodies, or other biological molecules or small molecules. Examples of biological immuno-oncology agents include, but are not limited to, cancer vaccines, antibodies, and cytokines. In one aspect, the antibody is a monoclonal antibody. In another aspect, the monoclonal antibody is a humanized or human antibody.

[0187] In one aspect, the immuno-oncology agent is either (i) a stimulatory (including costimulatory) receptor agonist or (ii) an antagonist of inhibitory (including costimulatory) signals on T cells, both of which result in amplification of antigen-specific T cell responses (often referred to as immune checkpoint modulators).

[0188] Suitable stimulatory and inhibitory molecules are members of the immunoglobulin superfamily (IgSF). One important family of membrane-bound ligands that bind to costimulatory or co-inhibitory receptors is the B7 family, which includes B7-1, B7-2, B7-H1 (PD-L1), B7-DC (PD-L2), B7-H2 (ICOS-L), B7-H3, B7-H4, B7-H5 (VISTA), and B7-H6. Another family of membrane-bound ligands that bind to costimulatory or coinhibitory receptors is CD40 and CD40L, OX-40, OX-40L, CD70, CD27L, CD30, CD30L, 4-1BBL, CD137 (4-1BB), TRAIL / Apo2-L, TRAILR1 / DR4, TRAILR2 / DR5, TRAILR3, TRAILR4, OPG, RANK, RANKL, TWEAKR / Fnl4, TWEAK, BAFFR, EDAR, XEDAR, TACI, APRIL, BCMA, LTpR, LIGHT, DcR3, HVEM, VEGETL1A, TRAMP / DR3, EDAR, EDA1, XEDAR, EDA2, TNFR1, lymphotoxin a / TNFp, TNFR2, TNFa, LTpR, and lymphotoxin a. The TNF family of molecules binds to cognate TNF receptor family members, including 1b2, FAS, FASL, RELT, DR6, TROY, and NGFR.

[0189] In one aspect, T cell responses can be stimulated by a combination of compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above with one or more of the following: (i) antagonists of proteins that inhibit T cell activation (e.g., immune checkpoint inhibitors), such as CTLA-4, PD-1, PD-L1, PD-L2, LAG-3, TIM-3, galectin-9, CEACAM-1, BTLA, CD69, galectin-1, TIGIT, CD113, GPR56, VISTA, 2B4, CD48, GARP, PD1H, LAIR1, TIM-1, and TIM-4, and (ii) Stimulators of proteins that stimulate T cell activation, such as B7-1, B7-2, CD28, 4-1BB (CD137), 4-1BBL, ICOS, ICOS-L, OX40, OX40L, GITR, GITRL, CD70, CD27, CD40, DR3, and CD28H.

[0190] Other drugs that can be used in combination with the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above for the treatment of cancer include antagonists of inhibitory receptors on NK cells or stimulators of activating receptors on NK cells. For example, the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above can be used in combination with KIR antagonists such as lirilumab.

[0191] Other agents for combination therapy include agents that inhibit or deplete macrophages or monocytes, including, but not limited to, CSF-1R antagonists such as CSF-1R antagonist antibodies, including RG7155.

[0192] Combination therapy is intended to encompass the administration of the therapeutic agents sequentially, i.e., administering each therapeutic agent at different times, as well as the administration of the therapeutic agent or at least two therapeutic agents substantially simultaneously.

[0193] Substantially simultaneous administration can be achieved, for example, by administering to a subject a single dosage form containing a fixed ratio of each therapeutic agent, or multiple single dosage forms of each therapeutic agent. Sequential or substantially simultaneous administration of each therapeutic agent can be achieved by any appropriate route of administration, including, but not limited to, oral administration, intravenous administration, intramuscular administration, and direct absorption through mucosal tissue. The therapeutic agents can be administered by the same route or by different routes. For example, the first therapeutic agent of a selected combination may be administered intravenously, while the other therapeutic agents of the combination may be administered orally. Alternatively, for example, all therapeutic agents may be administered orally, or all therapeutic agents may be administered intravenously. Combination therapy also includes further combination of the above-mentioned therapeutic agents with other biologically active ingredients and non-drug treatments (e.g., surgery or radiation therapy). When combination therapy further includes a non-drug treatment, the non-drug treatment may be administered at an appropriate time, as long as beneficial effects are obtained from the synergistic action of the combined therapeutic agent and non-drug treatment. For example, in appropriate cases, the non-drug treatment can be temporarily separated from the administration of the therapeutic agent, perhaps by days or weeks, and the beneficial effects still be achieved.

[0194] The types of cancer that may be treated with the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above include, but are not limited to, cancer of the prostate, colon, rectum, pancreas, cervix, stomach, endometrium, brain, liver, bladder, ovary, testis, head, neck, skin (including melanoma and basal cell carcinoma), mesothelial lining, white blood cells (including lymphoma and leukemia), esophagus, breast, muscle, connective tissue, lung (including small cell lung carcinoma and non-small cell carcinoma), adrenal gland, thyroid, kidney, or bone, or glioblastoma, mesothelioma, renal cell carcinoma, gastric cancer, sarcoma (including Kaposi's sarcoma), choriocarcinoma, basal cell carcinoma of the skin, hematological malignancies (including blood, bone marrow, and lymph nodes), or testicular seminoma.

[0195] To treat an eIF4A-associated disease, disorder, or condition, one or more additional pharmaceutical agents or therapies, for example, antiviral agents, chemotherapeutic or other anti-cancer agents, immunostimulants, immunosuppressants, radiation therapy, anti-tumor and anti-viral vaccines, cytokine therapy (e.g., IL2 and GM-CSF), and / or tyrosine kinase inhibitors, can optionally be used in combination with the compounds of Formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above. These agents can be combined with the compound in a single dosage form, or they can be administered simultaneously or sequentially in separate dosage forms.

[0196] Suitable chemotherapeutic or other anti-cancer agents include, for example, alkylating agents (including, but not limited to, nitrogen mustards, ethyleneimine derivatives, alkylsulfonates, nitrosoureas, and triazenes) such as uracil mustard, chlormethine, cyclophosphamide (CYTOXAN®), ifosfamide, melphalan, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, and temozolomide.

[0197] Agents suitable for use in combination with the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above in the treatment of melanoma include dacarbazine (DTIC), optionally in combination with other chemotherapeutic agents such as carmustine (BCNU) and cisplatin, the "Dartmouth therapy" consisting of DTIC, BCNU, cisplatin, and tamoxifen, and the combination of cisplatin, vinblastine, DTIC, temozolomide, or YERVOY™. The compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above may also be used in combination with immunotherapeutic agents, including cytokines such as interferon alpha, interleukin 2, and tumor necrosis factor (TNF), in the treatment of melanoma. The compounds of formula (I), (Ia), (Ib), (A), and (B) as defined above and a mixture of one enantiomer may be used in combination with vaccine therapy in the treatment of melanoma. Anti-melanoma vaccines are similar in some ways to antiviral vaccines used to prevent diseases caused by viruses such as polio, measles, and mumps. Weakened melanoma cells or parts of melanoma cells, called antigens, may be injected into patients to stimulate the body's immune system to destroy the melanoma cells.

[0198] Melanoma limited to the arms or legs may be treated using a combination of drugs, including one or more compounds of Formula (I), (Ia), (Ib), (A), and (B) as defined above, and one enantiomeric mixture, using hyperthermic isolated limb perfusion. In this treatment protocol, the blood circulation of the affected limb is temporarily isolated from the rest of the body, and high doses of chemotherapy drugs are infused into the arteries supplying that limb. This allows high doses to be delivered to the tumor area without exposing internal organs to these drugs, which can cause serious side effects. The fluid is usually warmed to 38.9°C to 40°C. Melphalan is the drug most commonly used in this chemotherapy. It can be administered with another drug called a tumor necrosis factor (TNF) inhibitor.

[0199] Suitable chemotherapeutic or other anti-cancer agents include, for example, antimetabolites (including, but not limited to, antifolates, pyrimidine analogs, purine analogs, and adenosine deaminase inhibitors) such as methotrexate, 5-fluorouracil, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, pentostatin, and gemcitabine.

[0200] Suitable chemotherapeutic or other anti-cancer agents further include certain natural products and their derivatives (e.g., vinca alkaloids, antitumor antibiotics, enzymes, lymphokines, and epipodophyllotoxins), such as, for example, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, ara-C, paclitaxel (taxol), mithramycin, deoxyco-formycin, mitomycin-C, L-asparaginase, interferons (especially IFN-α), etoposide, and teniposide.

[0201] Other cytotoxic agents include navelbene, CPT-11, anastrazole, letrazole, capecitabine, reloxafine, and droloxafine.

[0202] Also suitable are cytotoxic agents such as epidophyllotoxins, antitumor enzymes, topoisomerase inhibitors, procarbazine, mitoxantrone, platinum complexes such as cisplatin and carboplatin, biological response modifiers, growth inhibitors, antihormonal therapies, leucovorin, tegafur, and hematopoietic growth factors.

[0203] Other anti-cancer agents include antibody therapeutics such as trastuzumab (Herceptin®), antibodies against costimulatory molecules such as CTLA-4, 4-1BB and PD-1, or antibodies against cytokines (IL-1O or TGF-b).

[0204] Other anti-cancer drugs include those that block immune cell migration, such as antagonists to chemokine receptors including CCR2 and CCR4.

[0205] Other anti-cancer drugs include those that boost the immune system, such as adjuvants or adoptive T-cell transfer.

[0206] Anti-cancer vaccines include dendritic cells, synthetic peptides, DNA vaccines, and recombinant viruses.

[0207] In a specific embodiment of the present invention, at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and one enantiomeric mixture, and at least one chemotherapeutic agent are administered to a patient simultaneously or sequentially. In other words, at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and one enantiomeric mixture may be administered first, or at least one chemotherapeutic agent may be administered first, or at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and one enantiomeric mixture may be administered simultaneously. In addition, when two or more compounds of formula (I), (Ia), (Ib), (A), (B) as defined above, one enantiomeric mixture, and / or chemotherapeutic agents are used, these compounds may be administered in any order.

[0208] The present invention also provides pharmaceutical compositions comprising a therapeutically effective amount of one or more compounds of formula (I), (Ia), (Ib), (A), (B) as defined above, and one enantiomeric mixture, formulated together with one or more pharmaceutically acceptable carriers (excipients) and / or diluents, and optionally one or more additional therapeutic agents as described above.

[0209] The compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above may be administered by any suitable route, preferably in the form of a pharmaceutical composition adapted for such a route, in a dose effective for the intended treatment. The compounds of formula (I), (Ia), (Ib), (A), (B) and compositions thereof, as well as enantiomeric mixtures, as defined above, can be administered for any of the uses described herein by any suitable means, for example, oral administration such as tablets, capsules (including sustained-release or time-release formulations), pills, powders, granules, elixirs, tinctures, suspensions (including nanosuspensions, microsuspensions, and spray-dried dispersions), syrups, and emulsions; sublingual administration; buccal administration; parenteral administration such as subcutaneous, intravenous, intramuscular, or intrasternal injection, or infusion (e.g., sterile injectable aqueous or non-aqueous solutions or suspensions); nasal administration, including administration to the nasal mucosa, such as by inhalation spray; and topical administration, such as rectal administration in the form of a cream or ointment, or in the form of a suppository. They can be administered alone, but will usually be administered with a pharmaceutical carrier selected based on the chosen route of administration and standard pharmaceutical practice.

[0210] For oral administration, the pharmaceutical composition may be in the form of, for example, a tablet, capsule, liquid capsule, suspension, or liquid. The pharmaceutical composition is preferably prepared in the form of a dosage unit containing a specific amount of the active ingredient. For example, the pharmaceutical composition may be provided as a tablet or capsule containing about 0.1 to 1000 mg, preferably about 0.25 to 250 mg, and more preferably about 0.5 to 100 mg of the active ingredient. The appropriate daily dosage for humans or animals can vary greatly depending on the patient's condition and other factors, but can be determined using routine methods.

[0211] The pharmaceutical compositions contemplated herein can be orally administered, for example, via any acceptable and suitable oral formulation. Exemplary oral formulations include, but are not limited to, tablets, troches, lozenges, aqueous and oily suspensions, dispersible powders or granules, emulsions, hard and soft capsules, liquid capsules, syrups, and elixirs. Pharmaceutical compositions for oral administration can be prepared according to any method known in the art for preparing pharmaceutical compositions for oral administration. To provide a pharmaceutically palatable formulation, the pharmaceutical composition according to the present invention may contain at least one agent selected from sweeteners, flavoring agents, bittering agents, coloring agents, demulcents, antioxidants, and preservatives.

[0212] Tablets can be prepared, for example, by mixing at least one compound of formula (I), (Ia), (Ib), (A), or (B) as defined above and one enantiomeric mixture thereof, and / or at least one pharmaceutically acceptable salt thereof, with at least one non-toxic pharmaceutically acceptable excipient suitable for the manufacture of tablets. Exemplary excipients include, but are not limited to, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, sodium phosphate, etc.; granulating and disintegrating agents such as microcrystalline cellulose, croscarmellose sodium, corn starch, and alginic acid; binders such as starch, gelatin, polyvinylpyrrolidone, and acacia; and lubricants such as magnesium stearate, stearic acid, and talc. Tablets may also be uncoated or coated by known techniques to mask the unpleasant taste of unpleasant-tasting drugs or to delay disintegration and absorption of the active ingredient in the gastrointestinal tract, thereby prolonging the effect of the active ingredient. Examples of water soluble taste masking agents include, but are not limited to, hydroxypropyl methylcellulose and hydroxypropyl cellulose. Examples of time delay agents include, but are not limited to, ethyl cellulose and cellulose acetate butyrate.

[0213] Hard gelatin capsules can be prepared, for example, by mixing at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and one enantiomeric mixture, and / or at least one salt thereof, with at least one inert solid diluent, such as, for example, calcium carbonate, calcium phosphate, and kaolin.

[0214] Soft gelatin capsules can be prepared, for example, by mixing at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and one enantiomeric mixture, and / or at least one pharmaceutically acceptable salt thereof, with at least one water-soluble carrier, such as, for example, polyethylene glycol, and at least one oily medium, such as, for example, peanut oil, liquid paraffin, and olive oil.

[0215] Aqueous suspensions can be prepared, for example, by mixing at least one compound of formula (I), (Ia), (Ib), (A), (B) as defined above and one enantiomeric mixture, and / or at least one pharmaceutically acceptable salt thereof, and / or a prodrug or isotopically enriched form thereof, with at least one excipient suitable for the manufacture of aqueous suspensions. Exemplary excipients suitable for the manufacture of aqueous suspensions include, but are not limited to, suspending agents, such as sodium carboxymethylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, sodium alginate, alginic acid, polyvinylpyrrolidone, gum tragacanth, and gum acacia; dispersing or wetting agents, such as natural phospholipids such as lecithin; condensation products of alkylene oxides with fatty acids, such as polyoxyethylene stearate; condensation products of ethylene oxide with long chain aliphatic alcohols, such as heptadecaethyleneoxycetanol; condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols, such as polyoxyethylene sorbitol monooleate; and condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as polyethylene sorbitan monooleate. The aqueous suspensions may also contain at least one preservative (e.g., ethyl parahydroxybenzoate and n-propyl parahydroxybenzoate); at least one coloring agent; at least one flavoring agent; and / or at least one sweetening agent (including, but not limited to, sucrose, saccharin, and aspartame).

[0216] Oily suspensions can be prepared, for example, by suspending at least one compound of formula (I), (Ia), (Ib), (A), or (B) as defined above, and one enantiomeric mixture, and / or at least one pharmaceutically acceptable salt thereof, and / or a prodrug or isotopically enriched form thereof, in a vegetable oil such as peanut oil, olive oil, sesame oil, or coconut oil, or in a mineral oil such as liquid paraffin. Oily suspensions can also contain at least one thickening agent, such as beeswax, hard paraffin, and cetyl alcohol. To provide a palatable oily suspension, at least one sweetener and / or at least one flavoring agent, as already described above, can be added to the oily suspension. Oily suspensions can further contain at least one preservative, including, but not limited to, antioxidants such as butylated hydroxyanisole and α-tocopherol.

[0217] Dispersible powders and granules can be prepared, for example, by mixing at least one compound of formula (I), (Ia), (Ib), (A), or (B) as defined above, and one enantiomeric mixture thereof, and / or at least one pharmaceutically acceptable salt thereof, and / or a prodrug or isotopically enriched form thereof, with at least one dispersing agent and / or wetting agent, at least one suspending agent, and / or at least one preservative. Suitable dispersing agents, wetting agents, and suspending agents have already been described above. Examples of preservatives include, but are not limited to, antioxidants such as ascorbic acid. Dispersible powders and granules can also contain at least one excipient, including, but not limited to, sweeteners, flavoring agents, and coloring agents.

[0218] Emulsions of at least one compound of formula (I), (Ia), (Ib), (A), or (B) as defined above and one enantiomeric mixture thereof, and / or at least one pharmaceutically acceptable salt thereof, and / or a prodrug or isotopically enriched version thereof, can be prepared, for example, as an oil-in-water emulsion. The oil phase of the emulsion containing the compound of formula (I), (Ia), (Ib), (A), or (B) and the enantiomeric mixture as defined above can be composed of known ingredients in a known manner. This oil phase can be provided by, but is not limited to, vegetable oils such as olive oil and peanut oil, mineral oils such as liquid paraffin, and mixtures thereof. This phase can contain only an emulsifier, or it can contain a mixture of at least one emulsifier with a fat or oil, or both a fat and an oil. Suitable emulsifiers include, but are not limited to, naturally occurring phospholipids (e.g., soybean lecithin), esters or partial esters derived from fatty acids and hexitol anhydrides (e.g., sorbitan monooleate), and condensation products of partial esters with ethylene oxide (e.g., polyoxyethylene sorbitan monooleate). Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier that acts as a stabilizer. It is also preferred to include both an oil and a fat. The emulsifier, with or without a stabilizer, constitutes a so-called emulsifying wax, which, together with the oil and fat, constitutes a so-called emulsifying ointment base that forms the oily dispersed phase of a cream formulation. The emulsifier may also contain a sweetener, a flavoring, a preservative, and / or an antioxidant. Emulsifiers and emulsion stabilizers employed in the formulations of the present invention include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate, sodium lauryl sulfate, glyceryl distearate (alone or in combination with waxes), or other materials well known in the art.

[0219] The compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture thereof, and / or at least one pharmaceutically acceptable salt thereof, and / or prodrug or isotopically enriched thereof, as defined above, can also be delivered, for example, intravenously, subcutaneously, and / or intramuscularly, via any pharmaceutically acceptable and suitable injectable form. Examples of injectable forms include, but are not limited to, sterile aqueous solutions containing acceptable vehicles and solvents, such as water, Ringer's solution, and isotonic sodium chloride solution; sterile oil-in-water microemulsions, and aqueous or oily suspensions.

[0220] Formulations for parenteral administration may be in the form of isotonic sterile injection solutions or suspensions, such as aqueous or non-aqueous solutions. These solutions and suspensions can be prepared from sterile powders or granules using one or more of the carriers or diluents mentioned for oral formulations, or other suitable dispersing, wetting, or suspending agents. The compounds can be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, tragacanth gum, and / or various buffers. Other adjuvants and administration methods are well known and widely understood in the pharmaceutical arts. The active ingredient may be administered by injection in a composition with a suitable carrier, including saline, dextrose, and water, or in a composition with a cyclodextrin solubilizer (i.e., Captisol), a cosolvent solubilizer (i.e., propylene glycol), or a micelle solubilizer (i.e., Tween 80).

[0221] Sterile injectable preparations may also be sterile injectable solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, for example, as solutions in 1,3-butanediol. Among the acceptable vehicles and solvents that may be used are water, Ringer's solution, and isotonic sodium chloride solution. Sterile, solid oils are also conventionally used as solvents or suspending media. For this purpose, any bland, solid oil can be used, including synthetic mono- or diglycerides. Fatty acids, such as oleic acid, are also used in the preparation of injectables.

[0222] Sterile injectable oil-in-water microemulsions can be prepared, for example, by: 1) dissolving at least one compound of formula (I), (Ia), (Ib), (A), (B), and one enantiomeric mixture as defined above in an oil phase, such as a mixture of soybean oil and lecithin; 2) combining the compound of formula (I), (Ia), (Ib), (A), (B), and one enantiomeric mixture as defined above, comprising the oil phase, with a mixture of water and glycerol; and 3) treating the combination to form a microemulsion.

[0223] Sterile aqueous or oily suspensions can be prepared according to methods known in the art. For example, sterile aqueous solutions or suspensions can be prepared using non-toxic parenterally acceptable diluents or solvents, such as 1,3-butanediol, and sterile oily suspensions can be prepared using sterile, non-toxic acceptable solvents or suspending media, such as sterile solid oils, such as synthetic monoglycerides or diglycerides, and fatty acids, such as oleic acid.

[0224] Pharmaceutically acceptable carriers are formulated according to many factors within the understanding of those skilled in the art. These factors include, but are not limited to, the type and nature of the active agent being formulated, the subject to whom the drug containing the composition will be administered, the intended route of administration of the composition, and the targeted therapeutic indication. Pharmaceutically acceptable carriers include both aqueous and non-aqueous liquid media, as well as various solid and semi-solid dosage forms. Such carriers may contain many different components and additives in addition to the active agent. Such additional components are included in the formulation for various reasons well known to those skilled in the art, such as stabilization of the active agent or as a binder. Suitable pharmaceutically acceptable carriers and factors related to their selection are described in various available sources, such as Allen, LV Jr. et al., Remington: The Science and Practice of Pharmacy (Vol. 2), 22nd Edition (2012), Pharmaceutical Press.

[0225] Pharmaceutically acceptable carriers, adjuvants, and vehicles that can be used in the pharmaceutical compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS), such as d-α-tocopherol polyethylene glycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tween, polyethoxylated castor oil such as CREMOPHOR surfactants (BASF), or other similar polymeric delivery matrices, serum proteins, such as human serum albumin, buffer substances, such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol, and wool fat. Cyclodextrins, such as α-, β-, γ-cyclodextrin, or chemically modified derivatives, such as hydroxyalkyl cyclodextrins (including 2- and 3-hydroxypropyl-cyclodextrin), or other solubilizing derivatives, may also be advantageously used to enhance delivery of compounds of the formulae described herein.

[0226] The pharmaceutically active compounds of the present invention can be processed according to conventional pharmaceutical methods to produce medicaments for administration to patients, including humans and other mammals. The pharmaceutical compositions may be subjected to conventional pharmaceutical procedures such as sterilization and / or contain conventional adjuvants such as preservatives, stabilizers, wetting agents, emulsifiers, buffers, etc. Tablets and pills can also be prepared with enteric coatings. Such compositions may also contain adjuvants such as wetting agents, sweeteners, flavoring agents, and fragrances.

[0227] For therapeutic purposes, the active compound of the present invention is usually combined with one or more adjuvants suitable for the indicated administration route.For oral administration, the compound can be mixed with lactose, sucrose, starch powder, cellulose esters of alkanoic acid, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphate and sulfate, gelatin, gum arabic, sodium alginate, polyvinylpyrrolidone, and / or polyvinyl alcohol, and then tableted or encapsulated for easy administration.Such capsules or tablets can also contain controlled-release formulations, which can be provided in a dispersion of the active compound in hydroxypropylmethylcellulose.

[0228] The amount of compound administered and the dosage regimen for treating a disease state using the compounds and / or compositions of the present invention depend on various factors, including the age, weight, sex, and health of the subject, the type of disease, the severity of the disease, the route and frequency of administration, and the particular compound used. Thus, dosage regimens can vary widely but can be routinely determined using standard methods. A daily dosage of about 0.001 to 100 mg / kg body weight, preferably about 0.0025 to about 50 mg / kg body weight, and most preferably about 0.005 to 10 mg / kg body weight, may be appropriate. The daily dosage may be administered one to four times daily. Other dosage schedules include once-weekly administration and once-every-two-day administration.

[0229] The pharmaceutical compositions of the present invention comprise at least one compound of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above, and / or at least one pharmaceutically acceptable salt thereof and / or a prodrug or isotopically enriched form thereof, and optionally an additional agent selected from any pharmaceutically acceptable carrier, adjuvant, and vehicle. Alternative compositions of the present invention comprise a compound of formula (I), (Ia), (Ib), (A), (B) and an enantiomeric mixture as defined above, or a prodrug or isotopically enriched form thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

[0230] The present invention also includes pharmaceutical kits useful for treating or preventing, for example, RAS protein-related diseases. Accordingly, the present invention also relates to a kit comprising: a) a formulation comprising a pharmaceutical composition comprising one of the compounds of formula (I), (Ia), (Ib), (A), (B) and one of the enantiomeric mixtures defined above, or a therapeutically acceptable salt and / or prodrug or isotopically enriched form thereof, and a pharmaceutically acceptable carrier; and b) instructions for administering the pharmaceutical composition to treat a disease in which inhibition of RAS activation is effective in treating the disease.

[0231] Such kits may optionally further include one or more of a variety of conventional pharmaceutical kit components, such as containers containing one or more pharmaceutically acceptable carriers, additional containers as would be readily apparent to one of skill in the art, etc. Instructions indicating the amounts of components to be administered, administration guidelines, and / or guidelines for mixing the components may also be included in the kit, either as an insert or label.

[0232] It will be appreciated that the administration regimen for the compounds of the present invention will vary depending on known factors such as the pharmacodynamic properties of the particular drug, the method and route of administration, the patient's race, age, sex, health, condition and weight, the nature and extent of symptoms, type of concomitant treatment, frequency of treatment, route of administration, the patient's renal and hepatic function, and the desired effect.

[0233] As a general guideline, the daily oral dose of each active ingredient, when used to achieve the desired effect, will range from about 0.001 to about 5000 mg / day, preferably from about 0.01 to about 1000 mg / day, and most preferably from about 0.1 to about 250 mg / day. For intravenous administration, the most preferred dose will be in the range of about 0.01 to about 10 mg / kg / minute during a constant rate infusion. The compounds of Formula (I), (Ia), (Ib), (A), (B), and one enantiomeric mixture may be administered once daily, or the total daily dose may be divided into two, three, or four doses.

[0234] The compounds are typically administered in admixture with suitable pharmaceutical diluents, excipients, or carriers (collectively referred to herein as pharmaceutical carriers) appropriately selected according to the intended administration form, such as oral tablets, capsules, elixirs, and syrups, and in accordance with conventional pharmaceutical practice. Dosage forms (pharmaceutical compositions) suitable for administration may contain from about 1 mg to about 200 mg of active ingredient per dosage unit. In these pharmaceutical compositions, the active ingredient is usually present in an amount of from about 0.1 to 95% by weight, based on the total weight of the composition.

[0235] A typical capsule for oral administration contains at least one compound of Formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture (250 mg), lactose (75 mg), and magnesium stearate (15 mg), which mixture is passed through a 60-mesh sieve and filled into a No. 1 gelatin capsule.

[0236] A typical injection is prepared by aseptically placing at least one compound of Formula (I), (Ia), (Ib), (A), or (B) and one enantiomeric mixture (250 mg) into a vial, aseptically lyophilizing the vial, and sealing the vial. At the time of use, the contents of the vial are mixed with 2 mL of saline to prepare the injection.

[0237] The present invention includes within its scope pharmaceutical compositions comprising, as active ingredient, a therapeutically effective amount of at least one compound of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above, alone or in combination with a pharmaceutical carrier. Optionally, the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above can be used alone or in combination with other compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above, or in combination with one or more other therapeutic agents, such as anti-cancer agents or other pharmaceutically active substances.

[0238] Regardless of the route of administration selected, the compounds of formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture as defined above, which may be used in a suitable hydrated form, and / or the pharmaceutical compositions of the present invention, are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those skilled in the art.

[0239] Actual dosage levels of the active ingredients in the pharmaceutical compositions of the present invention may be varied so as to provide an amount of the active ingredient effective to achieve the desired therapeutic response for a particular patient, composition, and method of administration without causing toxicity to the patient.

[0240] The selected dosage level will depend upon a variety of factors, including the activity of the particular compound of formula (I), (Ia), (Ib), (A), (B) as defined above, or one enantiomeric mixture thereof, used, or an ester, salt, or amide thereof, the route of administration, the time of administration, the rate of excretion or metabolism of the particular compound used, the rate and extent of absorption, the duration of treatment, other drugs, compounds, and / or substances used in combination with the particular compound used, the age, sex, weight, condition, general health, and medical history of the patient being treated, and similar factors well known in the medical field.

[0241] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian can start the dosage of the compound of Formula (I), (Ia), (Ib), (A), (B) and one enantiomeric mixture used in the pharmaceutical composition as defined above at a lower amount than required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved.

[0242] Generally, a suitable daily dose of the compounds of formula (I), (Ia), (Ib), (A), and (B) and one enantiomeric mixture thereof as defined above will be the minimum dose effective to produce a therapeutic effect. Such an effective dose will generally depend on the factors described above. Generally, oral, intravenous, intracerebroventricular, and subcutaneous doses of the compounds of formula (I), (Ia), (Ib), (A), and (B) and one enantiomeric mixture thereof as defined above to a patient will range from about 0.01 to about 50 mg per kg of body weight per day.

[0243] If desired, the effective daily dose of the active compound may be administered as two, three, four, five, six, or more separate doses administered at appropriate intervals throughout the day, optionally in unit dosage form. In certain embodiments of the invention, administration is once daily, every other day, twice weekly, or once weekly.

[0244] While it is possible for the compounds of formula (I), (Ia), (Ib), (A), (B) and single enantiomeric mixtures as defined above to be administered alone, it is preferred to administer said compounds as pharmaceutical formulations (compositions).

[0245] The above-mentioned other therapeutic agents, when used in combination with the compounds of Formula (I), (Ia), (Ib), (A), (B) and single enantiomeric mixtures defined above, may be used, for example, in amounts indicated in the Physicians' Desk Reference (PDR) or in amounts determined by one skilled in the art. In the methods of the present invention, such other therapeutic agents may be administered before, simultaneously with, or after the administration of the compounds of the present invention.

[0246] The present invention will now be described in more detail with reference to the following examples, without the invention being limited to the particular embodiments described. The present invention includes all non-mutually exclusive combinations of the features described, particularly preferred features. [Brief explanation of the drawings]

[0247] Figure 1: MTT assay of cell viability in 96-well cell culture plates. RAS mutant cells were treated with Compounds 1 and C-1 for 48 hours, and cell viability was assessed by MTT assay. The results shown are the average of three independent experiments.

[0248] Figure 2: Effect of this compound on Ras activation NanoBiT assays of RAS GTP loading were performed in HeLa cells transfected with LgBit-K, N and HRAS G12V, and SmBit-CRAF-RBD. Cells were treated with compound 1 or C-1 for 2 hours in serum-free Dulbecco's modified Eagle's medium (DMEM). After incubation, NanoLuc substrate was added, and luminescence was measured using a multiplate reader. Data were normalized to cells transfected with the indicated mutants and exposed to DMSO for 2 hours. DMSO-treated cells were set to a value of 1. Results shown are the average of three independent experiments.

[0249] Figure 3: Dual-luciferase assay of cap-dependent translation initiation A dual-luciferase assay of cap-dependent translation initiation was performed in HeLa cells transfected with a pFR_HCV_xb-based dual-luciferase reporter gene. Cells were treated with Compound 1 or C-1 for 24 hours in serum-free Dulbecco's Modified Eagle's Medium (DMEM). After incubation, the dual-luciferase reporter assay was performed according to the manufacturer's instructions, and luminescence was measured using a multiplate reader. Data were normalized to transfected cells treated with DMSO for 24 hours. DMSO-treated cells were set as 1. Results shown are the average of three independent experiments.

[0250] Figure 4: Western blot analysis of JEKO cells treated with compound 1. JEKO cells (mantle cell lymphoma cell line) were treated with vehicle (DMSO) or compound 1 for various time points, and cell lysates were prepared. Proteins were separated by SDS-PAGE, and the expression of c-myc and cyclin D1 was analyzed by immunoblotting. Vinculin was used as a loading control.

[0251] Figure 5: Structures of compounds 1, 2, and C-1 [Example]

[0252] Abbreviation ACN Acetonitrile AIBN Azobis(isobutyronitrile) DCE Dichloroethane DMEM Dulbecco's Modified Eagle's Medium DCM dichloromethane DIPEA Diisopropylethylamine DMAP dimethylaminopyridine DMSO dimethyl sulfoxide FA formic acid FBS fetal bovine serum IPA Isopropyl Alcohol LDA Lithium diisopropylamide NBS N-Bromosuccinimide PBS Phosphate-buffered saline PE PET Ether RLU Relative Luminescence Units (Luciferase Activity) / Relative Luciferase Units SD standard deviation SEM Trimethylsilylethoxymethyl TBAF Tetra-n-butylammonium fluoride TFA trifluoroacetic acid THF tetrahydrofuran TLC thin layer chromatography TMSCN Trimethylsilyl cyanide Triton B Benzyltrimethylammonium Hydroxide

[0253] Materials and Methods cell: HCT-116 cells were authenticated by Eurofin Genomics and cultured in DMEM (10% heat-inactivated FBS). sNF96.2 cells were purchased from ATCC and cultured in DMEM (10% heat-inactivated FBS + 1 mM Na-pyruvate). ASPC-1 cells were purchased from DSMZ and cultured in RPMI-1640 (10% heat-inactivated FBS). NCI-H358 cells were purchased from ATCC and cultured in RPMI-1640 (10% heat-inactivated FBS). T24 and HCT-116 cells were cultured in McCoy's 5A medium (10% heat-inactivated FBS). HT-1080 cells were cultured in EMEM (10% heat-inactivated FBS + 1 mM Na-pyruvate).

[0254] DNA transfection: HeLa cells were harvested with 0.05% trypsin / 0.02% EDTA in PBS and plated in 6- or 12-well cell culture plates at 5 × 10 4 Cells were seeded in complete DMEM (2 ml for 6-well plates and 1 ml for 12-well plates) at a concentration of 0.1 mL. One day after seeding, Flag-tagged Ras or NanoBiT plasmids were transfected into the cells using a PEI / PBS solution.

[0255] NanoBit Assay: The plasmid was transfected into HeLa cells, which were harvested one day after transfection. The cells were seeded into a 96-well white plate. The next day, the medium was replaced with serum-free DMEM and the cells were incubated with the compounds for 2 hours. After incubation, a Nano Glo assay was performed. Luminescence was measured using a Tecan SPARKS (Tecan). IC50 values ​​were calculated using nonlinear regression analysis with GraphPad Prism 9.

[0256] Dual-luciferase assay: 1 × 10 HeLa cells 6Cells were seeded at 2 ml / ml in growth medium into 6-well cell culture plates. 2 μg of plasmid was transfected into cells using 0.5 mM PEI reagent dissolved in 200 μl of PBS. The day after transfection, cells were harvested and seeded into half of a 96-well white plate (Greiner). After another day, the medium was replaced with serum-free DMEM, and the cells were incubated with the compounds for 24 hours. Dual-Glo luciferase assays were performed according to the manufacturer's instructions (Promega, N2920). Luminescence was measured using a Tecan infinite (Tecan). IC50 values ​​were calculated using GraphPad Prism 9 by nonlinear regression.

[0257] MTT assay: 5 × 10 cancer cells 3 Cells were seeded in 80 μl of growth medium per well of a 96-well plate and cultured in an incubator for 1 day. The next day, 20 μl of growth medium containing compound was added to each well, and the cells were cultured for 48 hours. After incubation with compound, 10 μl of MTT solution was added to the well and incubated for 2–4 hours. After incubation with MTT, solubilization buffer was added and the cells were incubated overnight. MTT was measured at OD570 nm using a plate reader (Tecan). IC50 values ​​were calculated using nonlinear regression analysis using GraphPad Prism 9.

[0258] Synthesis: Compound A: 4-((1R,3S,3aR,8bS)-3-(3-fluorophenyl)-1,8b-dihydroxy-6,8-dimethoxy-1,2,3,8b-tetrahydro-3aH-cyclopenta[b]benzofuran-3a-yl)benzonitrile JPEG2026508161000012.jpg126117

[0259] compound 2 To a solution of compound 1 (50 g, 219.3 mmol, 1 equiv.) in DCE (500 mL) at 25 °C, AIBN (3.23 g, 19.64 mmol, 0.09 equiv.) and NBS (46.62 g, 261.93 mmol, 1.2 equiv.) were added. The mixture was then stirred at 65 °C for 16 h under a nitrogen atmosphere. TLC (petroleum ether / ethyl acetate = 10 / 1, R f=0.8), indicating that compound 1 was completely consumed, and one major peak of the desired mass was detected. The reaction mixture was quenched with water (500 mL) at 25 °C and extracted with DCM (1 L × 2). The combined organic layers were washed with brine (500 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give compound 2 (74 g, crude) as a white solid, which was used in the next step without further purification.

[0260] compound 4 To a solution of crude compound 2 (60 g, approximately 176.5 mmol, 90% purity, 1 equivalent) in acetone (420 mL) at 25 °C, K2CO3 (41.20 g, 298.08 mmol, 1.7 equivalents) and 3,5-dimethoxyphenol (30.00 g, 194.63 mmol, 1.11 equivalents) were added. The mixture was then stirred at 70 °C under a nitrogen atmosphere for 16 hours. TLC (petroleum ether / ethyl acetate = 10 / 1, R f =0.60), indicating the complete consumption of compound 2 and the formation of one new spot. The reaction mixture was filtered, concentrated, diluted with HO, and extracted three times with EtOAc. The combined organic layers were washed with brine (500 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give crude compound 4. The residue was purified by column chromatography (SiO, petroleum ether / ethyl acetate = 100 / 1 to 5 / 1) to give compound 4 (57.6 g, 151.6 mmol, 86% yield for two steps) as a white solid. 1 H NMR (400 MHz, CDCl3)δ = 7.59 - 7.50(m, 2H), 7.49 - 7.41(m, 2H), 6.11(s, 3H), 5.57(s, 1H), 3.82 - 3.70(m, 9H).

[0261] compound 5 To a solution of compound 4 (60 g, 157.9 mmol, 1 equiv.) in MeOH (420 mL) and HO (42 mL) was added KCO (26.10 g, 188.87 mmol, 1.2 equiv.) at 25 °C. The mixture was stirred at 50 °C under a nitrogen atmosphere for 16 h. TLC (petroleum ether / ethyl acetate = 1:1, R f=0.60), indicating the complete consumption of compound 4 and the formation of one new spot. Four parallel reactions were worked up together. The reaction mixture was quenched with water (1 L) at 25 °C, and then the pH of the reaction was adjusted with HCl (6 mol / L) and extracted with EtOAc (1 L × 3). The combined organic layers were washed with brine (1 L), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was then triturated with petroleum ether / ethyl acetate = 20:1, filtered, and concentrated under reduced pressure to give compound 5 (325 g, 888.0 mmol, 71%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ = 7.60 - 7.50 (m, 2H), 7.49 - 7.40 (m, 2H), 6.21 - 6.01 (m, 3H), 5.58 (s, 1H), 3.75 (s, 6H).

[0262] compound 6 To a mixture of compound 5 (25 g, 68.3 mmol, 1 equiv.) and ZnCl2 (12.99 g, 95.32 mmol, 4.46 mL, 1.4 equiv.) at 25 °C, POCl3 (100 mL) was added, and the mixture was stirred at 25 °C for 16 h. TLC (petroleum ether / ethyl acetate = 2:1, R f =0.50), indicating the complete consumption of compound 5 and the formation of one new spot. Four parallel reactions were worked up together. The reaction mixture was quenched by slow dropwise addition to water (1000 mL) at 25–40 °C and filtered to give a residue. The residue was then washed with water (three times), filtered, and concentrated under reduced pressure to give the crude product. The residue was then triturated with methyl alcohol, filtered, and concentrated under reduced pressure to give compound 6 (83 g, 238.50 mmol, 87% yield) as a white solid. 1 H NMR (400 MHz, CDCl3) δ = 7.53 - 7.48 (m, 2H), 7.31 (d, J = 8.3 Hz, 2H), 6.27 (d, J = 1.9 Hz, 1H), 6.06 (d, J = 1.8 Hz, 1H), 5.43 (s, 1H), 3.91 (d, J = 2.0Hz, 6H).

[0263] compound 8 To a solution of compound 6 (10 g, 28.7 mmol, 1 equiv.) in t-BuOH (360 mL) at 25 °C, Triton B (1.20 g, 2.86 mmol, 1.30 mL, 40% purity, 0.1 equiv.) and compound 7 (5.16 g, 34.37 mmol, 1.2 equiv.) in t-BuOH (40 mL) were added. The mixture was stirred at 60 °C under a nitrogen atmosphere for 2 h. LCMS showed that compound 6 was completely consumed, and two peaks corresponding to the desired mass were detected. The solvent was removed in vacuo, and the mixture was quenched with water (50 mL) and acidified to pH = 1 with 6 M HCl. The residue was extracted with ethyl acetate (100 mL × 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give compound 8 (3.01 g, 6.0 mmol, 21% yield) as a white solid and its diastereomer 8A (1.32 g, 2.65 mmol, 9% yield) as a white solid. Compound 8: 1 H NMR(400 MHz, CDCl3)δ = 9.44(d, J = 1.1 Hz, 1H), 7.67 - 7.58(m, 2H), 7.54 - 7.44(m, 2H), 7.17 - 7.09(m, 2H), 7.06 - 6.99(m, 1H), 6.84 - 6.76(m, 1H), 6.24(d, J = 1.9 Hz, 1H), 5.85(d, J = 1.8 Hz, 1H), 4.20(dd, J = 3.8, 10.8 Hz, 1H), 3.87(s, 3H), 3.73(s, 3H), 3.06(ddd, J = 2.1, 10.9, 17.7 Hz, 1H), 2.75 - 2.51(m, 1H). Compound 8A: 1H NMR (400 MHz, CDCl3) δ = 9.51 (d, J = 2.1 Hz, 1H), 7.39 - 7.34 (m, 2H), 7.32 - 7.28 (m, 2H), 7.09 (dt, J = 6.1, 7.9 Hz, 1H), 7.02 - 6.96 (m, 1H), 6.89 (d, J = 7.9 Hz, 1H), 6.82 (dt, J = 2.6, 8.3 Hz, 1H), 6.37 (d, J = 1.8 Hz, 1H), 6.05 (d, J = 1.8 Hz, 1H), 4.16 (dd, J = 3.4, 11.1 Hz, 1H), 3.94 (s, 3H), 3.89 (s, 3H), 2.97 (ddd, J = 2.5, 11.0, 17.2 Hz, 1H), 2.65 (dd, J = 3.4, 17.3 Hz, 1H).

[0264] compound 9 To a solution of compound 8 (22.5 g, 45.2 mmol, 1 equiv.) in CH3CN (225 mL) at 25 °C, TMSCN (14.31 g, 144.19 mmol, 18.04 mL, 3.2 equiv.) and ZnI2 (1.44 g, 4.51 mmol, 0.1 equiv.) were added. The mixture was stirred under a nitrogen atmosphere at 25 °C for 16 h. LCMS showed that compound 8 was completely consumed, and one major peak of the desired mass was detected. The reaction mixture was quenched with water (200 mL) and extracted with EtOAc (150 mL × 2). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give compound 9 (27.15 g, crude) as a yellow solid. The residue was used in the next step without further purification. 1H NMR (400 MHz, CDCl3) δ = 7.63 - 7.55 (m, 2H), 7.54 - 7.49 (m, 2H), 7.23 - 7.10 (m, 2H), 7.10 - 6.95 (m, 2H), 6.91 - 6.82 (m, 1H), 6.26 (t, J = 2.2 Hz, 1H), 5.85 (dd, J = 1.9, 3.0 Hz, 1H), 3.88 (s, 4H), 3.72 (d, J = 1.8 Hz, 3H), 2.01 (s, 3H), 0.05 (d, J = 3.8 Hz, 9H).

[0265] compound 11 To a solution of compound 9 (21.6 g, 36.2 mmol, 1 equiv.) in THF (432 mL) was added dropwise LDA (2 M, 54.13 mL, 3 equiv.) at -78 °C. The mixture was stirred under a nitrogen atmosphere at -78 °C for 1 h. LCMS showed that compound 9 was completely consumed, and one major peak of the desired mass was detected. The reaction mixture was quenched with water / ice (500 mL) and extracted with EtOAc (500 mL × 2). The combined organic layers were washed with brine (500 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give compound 11 (14.2 g, 23.7 mmol, 66% yield) as a yellow solid. INT.10 (2.5 g, 5.0 mmol, 14% yield) was also obtained as a yellow solid. Compound 11 1H NMR (400 MHz, CDCl3) δ = 7.29 (br d, J = 8.4 Hz, 1H), 7.20 (d, J = 8.3 Hz, 2H), 7.11 (q, J = 7.3 Hz, 1H), 7.01 - 6.94 (m, 2H), 6.85 (d, J = 7.8 Hz, 1H), 6.82 - 6.73 (m, 2H), 6.21 (s, 1H), 6.05 (s, 1H), 4.32 (dd, J = 5.6, 14.4 Hz, 1H), 3.85 (s, 3H), 3.82 (s, 3H), 3.17 - 2.99 (m, 2H), 2.68 (dd, J=5.7, 12.7 Hz, 1H), 0.11(s, 9H).

[0266] INT.10 To a solution of compound 11 (14.2 g, 23.8 mmol, 1 equiv.) in THF (142 mL) at 0 °C, TBAF (1 M, 29.66 mL, 1.25 equiv.) was added. The mixture was stirred under a nitrogen atmosphere at 25 °C for 2 h. LCMS showed complete consumption of compound 11, and one major peak of the desired mass was detected. The reaction mixture was quenched with water (150 mL) and extracted with EtOAc (150 mL × 2). The combined organic layers were washed with brine (150 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give INT.10 (10.2 g, 20.5 mmol, 86% yield) as a yellow solid. LCMS: M+23 (520.9), 92.7%. 1H NMR (400 MHz, CDCl3) δ = 7.35 - 7.28 (m, 2H), 7.15 - 7.03 (m, 2H), 6.95 (d, J = 8.6 Hz, 2H), 6.83 (br t, J = 8.1 Hz, 1H), 6.74 (br d, J = 8.1 Hz, 2H), 6.36 (d, J = 1.8 Hz, 1H), 6.13 (d, J = 1.7 Hz, 1H), 5.31 (s, 1H), 3.97 - 3.91 (m, 1H), 3.87 (s, 3H), 3.85 (s, 1H), 3.83 (s, 3H), 3.29 (br s, 3.13 - 3.02(m, 1H), 3.02 - 2.90(m, 1H).

[0267] compound 12 To a mixture of INT.10 (300 mg, 0.60 mmol, 1 equiv.) in DMF (6 mL) at 20 °C, Zn(CN) (282.19 mg, 2.40 mmol, 152.53 μL, 4 equiv.), Pd(dba) (55.02 mg, 60.08 μmol, 0.1 equiv.), and DPPF (66.62 mg, 120.16 μmol, 0.2 equiv.) were added under a nitrogen atmosphere. The mixture was stirred at 120 °C for 16 h. LCMS showed that INT.10 had been consumed and the desired product was the major component. At 25 °C, the reaction mixture was quenched with water (120 mL) and extracted with EtOAc (100 mL × 2). The combined organic layers were washed with brine (100 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by preparative MPLC (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 16 / 1) to give compound 12 (210 mg, 0.47 mmol, 78% yield) as a yellow solid. LCMS: M+1 (446.1), 80.1%. 1H NMR (400 MHz, CDCl3) δ = 7.46 (d, J = 8.4 Hz, 2H), 7.18 (d, J = 8.4 Hz, 2H), 7.12 - 7.05 (m, 1H), 6.86 - 6.78 (m, 1H), 6.72 (br t, J = 7.0 Hz, 2H), 6.37 (d, J = 1.6 Hz, 1H), 6.14 (d, J = 1.4 Hz, 1H), 4.01 - 3.92 (m, 1H), 3.87 (s, 3H), 3.84 (s, 3H), 3.19 - 3.08 (m, 1H), 3.01 (br d, J = 12.3 Hz, 1H).

[0268] compound 13 To a solution of compound 12 (0.23 g, 516.35 μmol, 1 equiv.) in MeCN (4.5 mL) and AcOH (0.5 mL) was added NaBH(OAc) (2.19 g, 10.33 mmol, 20 equiv.) at 25° C. The mixture was then stirred at 50° C. for 16 h. LCMS showed complete consumption of the starting material, and one major peak of the desired mass was detected. The reaction mixture was quenched with water (30 mL) at 25° C. and extracted with EtOAc (30 mL×2). The combined organic layers were washed with brine (20 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex luna C18 100 × 40 mm, 3 μm; mobile phase: [HO (0.1% TFA)-ACN]; gradient: 30% to 60% B over 8.0 min) to give compound 13 (0.1 g, 223.49 μmol, 43% yield) as a yellow solid. LCMS: M+1 (448.15), 70.5%. 11H NMR (400 MHz, CDCl3) δ = 7.47 - 7.40 (m, 2H), 7.38 - 7.31 (m, 2H), 7.11 - 7.01 (m, 1H), 6.83 - 6.76 (m, 1H), 6.76 - 6.69 (m, 2H), 6.31 (d, J = 1.8 Hz, 1H), 6.18 (d, J = 1.8 Hz, 1H), 4.82 (d, J = 5.8 Hz, 1H), 4.10 - 4.03 (m, 1H), 3.91 (s, 3H), 3.86 (s, 3H), 2.73 (dt, J = 6.2, 13.9 Hz, 1H), 2.24(dd, J = 6.5, 13.6 Hz, 1H).

[0269] Compound A Compound 13 (0.1 g, 223.49 μmol, 1 equivalent) was purified by SFC (column: DAICEL CHIRALPAK AD (250 mm × 30 mm, 10 μm); mobile phase: [CO2 - IPA]; B%: 45%, isocratic elution mode), and two peaks were obtained. Peak 1, Compound A (20.7 mg, 46.3 μmol) white solid. LCMS: M + 23 (470.1), 97.8%. 1 1H NMR (400 MHz, CDCl3) δ = 7.48 - 7.40 (m, 2H), 7.39 - 7.32 (m, 2H), 7.12 - 7.02 (m, 1H), 6.84 - 6.68 (m, 3H), 6.32 (s, 1H), 6.18 (s, 1H), 4.83 (br d, J = 6.1 Hz, 1H), 4.08 (br dd, J = 6.4, 14.1 Hz, 1H), 3.92 (s, 3H), 3.87 (s, 3H), 3.12 (s, 1H), 2.73 (dt, J = 6.1, 14.0 Hz, 1H), 2.25 (dd, J = 6.3, 13.6 Hz, 1H), 1.82(s, 1H). Peak 2, enantiomer of Compound A (21.7 mg, 48.50 μmol) white solid. LCMS: M + 23 (470.1), 99%. 1H NMR (400 MHz, CDCl3) δ = 7.44 (br d, J = 8.4 Hz, 2H), 7.35 (br d, J = 8.4 Hz, 2H), 7.07 (br d, J = 6.4 Hz, 1H), 6.88 - 6.68 (m, 3H), 6.32 (d, J = 1.0 Hz, 1H), 6.18 (s, 1H), 4.83 (br d, J = 5.8 Hz, 1H), 4.08 (br dd, J = 6.6, 13.9 Hz, 1H), 3.92 (s, 3H), 3.87 (s, 3H), 3.12 (s, 1H), 2.73 (dt, J = 6.1, 13.7 Hz, 1H), 2.25(br dd, J = 6.8, 13.9 Hz, 1H), 1.82(s, 1H).

[0270] Compound B: 4-((1R,3S,3aR,8bS)-1,8b-dihydroxy-6,8-dimethoxy-3-phenyl-1,2,3,8b-tetrahydro-3aH-cyclopenta[b]benzofuran-3a-yl)benzonitrile JPEG2026508161000013.jpg44117

[0271] compound 15 To a solution of (CH)N(CHCO)BH (273 mg, 1.04 mmol) in acetonitrile (2 mL) was added AcOH (1 mL) at 0 °C and stirred for 30 min at room temperature. Then, a solution of (3S,3aR,8bR)-3a-(4-bromophenyl)-8b-hydroxy-6,8-dimethoxy-3-phenyl-2,3,3a,8b-tetrahydro-1H-cyclopenta[b]benzofuran-1-one (14) (50 mg, 0.10 mmol) in acetonitrile (2 mL) was added dropwise at 0 °C, and the reaction mixture was stirred at room temperature for 7 h. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with dilute HCl (1.5 N, 4 mL) at 0 °C, and the aqueous layer was extracted with DCM (30 mL × 2). The combined organic layers were washed with brine (4 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated under reduced pressure, and the crude compound obtained was purified by flash column chromatography (silica gel: 230-400 mesh, eluent: 10-20% ethyl acetate / petroleum ether gradient) to give compound 15 (40 mg, 80%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6): δ 7.21 (d, J = 11.6 Hz, 2H), 7.12-7.09 (m, 2H), 7.05-7.02 (m, 5H), 6.27 (d, J = 2.0 Hz, 1H), 6.12 (d, J = 2.0 Hz, 1H), 4.98 (s, 1H), 4.50-4.49 (m, 1H), 4.47-4.45 (m, 1H), 3.80 (dd, J = 5.6, 14.0 Hz, 1H), 3.78 (s, 3H), 3.74 (s, 3H), 2.71-2.67 (m, 1H), 2.02 (dd, J = 6.4, 12.8 Hz, 1H).

[0272] compound 16 To a solution of (1R,3S,3aR,8bS)-3a-(4-bromophenyl)-6,8-dimethoxy-3-phenyl-1,2,3,3a-tetrahydro-8bH-cyclopenta[b]benzofuran-1,8b-diol (35 mg, 0.07 mmol) in DMF (2 mL) was added zinc cyanide (50.8 mg, 0.43 mmol), followed by zinc powder (0.47 mg, 7.24 μmol). The reaction mixture was degassed with nitrogen gas for 5 min, and then dppf (8.03 mg, 0.01 mmol) was added at room temperature, followed by Pd2(dba)3 (6.63 mg, 7.24 μmol). The reaction mixture was stirred at 100 °C for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through a pad of Celite and washed with EtOAc (10 mL) followed by water (4 mL). The combined organic layers were extracted with EtOAc (20 mL), and the organic portion was washed with brine (5 mL) and dried over anhydrous Na2SO4. The organic portion was filtered and concentrated in vacuo. The crude product was purified by preparative HPLC to give compound 16 (12 mg, 39% yield) as an off-white solid. LCMS: 412.2 (M+ -OH), 1 H-NMR (400 MHz, DMSO-d6): δ 7.50 (d, J = 8.4 Hz, 2H), 7.27 (d, J = 8.4 Hz, 2H),7.12-7.08 (m, 2H), 7.04-7.00 (m, 3H), 6.29 (d, J = 2.0 Hz, 1H), 6.13 (d, J = 2.0 Hz, 1H), 5.10 (s, 1H), 4.58 (d, J = 3.6 Hz, 1H), 4.47-4.45 (m, 1H), 4.08 (dd, J = 6.4, 14.8 Hz, 1H), 3.78 (s, 3H), 3.74(s, 3H), 2.77-2.76(m, 1H), 2.09(dd, J = 5.6, 12.8 Hz, 1H).

[0273] Compound B Compound 16 (20 mg, 0.047 mmol) was purified by chiral SFC (CO₂:methanol 60:40 to 80:20, flow rate: 3 mL / min, column: YMC Cellulose-SJ). The first peak was concentrated in vacuo at 45 °C to give ent-B. Compound B was obtained from the second peak.

[0274] Compound B enantiomer: 9 mg, off-white solid. LCMS: 412.2 (M+-OH). 1 H-NMR (400 MHz, DMSO-d6): δ 7.50 (d, J = 8.4 Hz, 2H), 7.27 (d, J = 8.4 Hz, 2H), 7.11-7.08 (m, 2H), 7.04-7.00 (m, 3H), 6.29 (d, J = 2.0 Hz, 1H), 6.13 (d, J = 2.0 Hz, 1H), 5.10 (s, 1H), 4.58 (d, J = 3.6 Hz, 1H), 4.47-4.45 (m, 1H), 4.08 (dd, J = 6.4, 14.8 Hz, 1H), 3.78 (s, 3H), 3.74(s, 3H), 2.81-2.73(m, 1H), 2.07(dd, J = 5.6, 12.8 Hz, 1H).

[0275] Compound B: 6 mg, off-white solid. LCMS: 430.0 (M++H). 1H-NMR (400 MHz, DMSO-d6): δ 7.50 (d, J = 8.4 Hz, 2H), 7.27 (d, J = 8.4 Hz, 2H), 7.12-7.08 (m, 2H), 7.04-7.02 (m, 3H), 6.29 (d, J = 2.0 Hz, 1H), 6.13 (d, J = 2.0 Hz, 1H), 5.10 (s, 1H), 4.58 (d, J = 3.6 Hz, 1H), 4.47-4.45 (m, 1H), 4.08 (dd, J = 6.4, 14.8 Hz, 1H), 3.78 (s, 3H), 3.74 (s, 3H), 2.77-2.74 (m, 1H), 2.08 (dd, J = 5.6, 12.8 Hz, 1H).

[0276] Compound C-1 (1R,3S,3aR,8bS)-3a-(4-chlorophenyl)-6,8-dimethoxy-3-phenyl-1,2,3,3a-tetrahydro-8bH-cyclopenta[b]benzofuran-1,8b-diol (N. Ribeiro et al., J. Med. Chem., 2012, 55, 100064) was synthesized in a similar manner. JPEG2026508161000014.jpg95141LCMS: 439.2(M++H). 1 H NMR (400 MHz, DMSO-d6): δ 7.10-7.08 (m, 6H), 7.03-7.01 (m, 3H), 6.26 (d, J = 2.0 Hz, 1H), 6.13 (d, J = 2.0 Hz, 1H), 4.98 (s, 1H), 4.49-4.48 (m, 1H), 4.47-4.46 (m, 1H), 4.00 (dd, J = 5.6, 6.4 Hz, 1H), 3.78 (s, 3H), 3.75 (s, 3H), 2.75-2.70 (m, 1H), 2.05-2.00 (m, 1H).

[0277] result: JPEG2026508161000015.jpg249167JPEG2026508161000016.jpg27166

[0278] Unbound fraction in human plasma and pharmacokinetics (Tables 1 and 3) After administration, a certain amount of drug is present in the blood or plasma. Of the total amount in the blood or plasma, a portion is bound to proteins (or other molecular species in the blood or plasma). Only the unbound drug is available for drug-target interactions, etc. Therefore, the more unbound fraction available in the blood or plasma, the more drug is available to bind to the target. The less unbound drug in the blood or plasma (e.g., due to being involved in competitive binding with albumin), the less available for the desired drug-target interaction. Protocols for measuring the unbound fraction and pharmacokinetics are routine and well known to those skilled in the art.

[0279] Surprisingly, the compounds of the present invention have a much higher free fraction in human plasma, meaning that a much higher amount of drug is available to bind to the target compared to prior art compounds.

[0280] The human microsomal clearance (hMic Clint) of the compounds of the present invention is lower compared to C-1. The exposure of a drug (AUC, area under the concentration-time curve) depends, inter alia, on its clearance (the lower the clearance of a drug, the higher the exposure). Generally, the beneficial effect of a drug depends on its potency and exposure, so the lower the clearance, the lower the dose required to achieve the same beneficial effect. AUC = (FD) / CL In the formula, F is the bioavailability (100% for intravenous administration), D is the dose, and CL is the clearance.

[0281] Table 1: PK comparison of three related compounds at 2 mg / kg iv (intravenous). In conclusion, the free AUC and Cave are higher for compounds 1 and 2 than for compound C-1. JPEG2026508161000017.jpg76166

[0282] Table 2: Comparison of oral PK at 50 mg / kg oral administration of C-1 and 1. In conclusion, the free AUC and Cave of 1 were higher than those of C-1. JPEG2026508161000018.jpg69164

[0283] Table 3: Comparison of oral PK at 10 mg / kg oral administration of C-1 and 2. In conclusion, the free AUC and Cave were higher for 2 than for C-1. JPEG2026508161000019.jpg69163

[0284] Table 4: This table summarizes the unbound clearance data. In conclusion, 1 and 2 have lower unbound clearance, hence their higher unbound AUC. JPEG2026508161000020.jpg61164

[0285] Unbound Fraction Protocol Dialysis machine: HT dialysis plates (model HTD 96 b, catalog number 1006) and dialysis membranes (molecular weight cutoff 12-14 kDa, catalog number 1101) were purchased from HT Dialysis LLC (Gales Ferry, CT). Dialysis buffer (100 mM sodium phosphate and 150 mM NaCl, pH 7.4±0.1) Stop solution (tolbutamide (200 ng / mL) and labetalol (200 ng / mL) in acetonitrile)

[0286] Dialysis membrane and matrix processing procedure: On the day of the experiment, the plasma was thawed under running water and centrifuged at 3220 × g for 5 minutes to remove clots. The pH value was confirmed and recorded. Only plasma with a pH between 7.0 and 8.0 was used.

[0287] Dialysis membranes were pretreated according to the manufacturer's instructions. Dialysis membrane strips were immersed in ultrapure water at room temperature for approximately 1 hour. Then, each strip containing two membranes was separated and immersed in ethanol:water (20:80 v:v) for approximately 20 minutes. They were then ready to use or stored in the solution at 2-8°C for up to 1 month. Before use, the membranes were washed and immersed in ultrapure water for 20 minutes.

[0288] Test and control compound dilution procedure: A working solution (400 μM) of the test compound and the control compound was prepared. 3 μL of the working solution was added to 597 μL of blank matrix to achieve the final concentration, and the mixture was mixed well.

[0289] Assay procedure 50 μL of loading matrix containing the test compound or control compound was transferred to a sample collection plate in triplicate. The sample was immediately mixed with the blank buffer in the opposite well to achieve a 1:1 (v:v) matrix:dialysis buffer volume ratio in each well, resulting in a final volume of 100 μL. A stop solution was added to these TO samples of test and control compounds. The plate was sealed and shaken at 800 rpm for 10 minutes. These TO samples, along with the other dialyzed samples, were then stored at 2–8°C until further processing.

[0290] The dialysis apparatus was assembled according to the manufacturer's instructions. 100 μL of loading matrix containing test or control compounds was dispensed in triplicate into the donor side of each dialysis well, and 100 μL of dialysis buffer was loaded into the receiver side.The plate was then rotated at approximately 100 rpm in a humidified incubator at 37±1°C with 5% CO for 4 hours.

[0291] At the end of dialysis, 50 μL samples were collected from the buffer and matrix sides of the dialyzer into a new 96-well plate (sample collection plate). An equal volume of blank matrix (buffer or matrix) was added to each sample to bring the final volume to 100 μL. The volume ratio of matrix to dialysis buffer in each well was 1:1 (v:v). Protein precipitation was performed on all samples for LC-MS / MS analysis. Detailed sample processing methods are described in the study record.

[0292] Data Analysis: The % unbound, % bound and % recovery were calculated using the following formulas: %unbonded = 100 × F / T %bound = 100 - %unbound % Recovery = 100 × (F + T) / T0 F = free compound concentration measured at the calculated concentration on the buffer side of the membrane T = total compound concentration measured at the calculated concentration on the matrix side of the membrane T0 = ​​total compound concentration measured at the calculated concentration in the matrix before dialysis

Claims

1. Compounds of formula (I) (In the formula, R 1 is CN and R 2 is selected from hydrogen and F), a prodrug or isotopically enriched thereof, or a pharmaceutically acceptable salt thereof.

2. Compounds of formula (Ia) or (Ib) (In the formula, R 1 and R 2 has the same meaning as defined in claim 1), or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof.

3. 10. A compound of formula (I) according to any preceding claim, which is a mixture of (Ia) and (Ib), a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, wherein the enantiomeric excess (ee) of the enantiomer of formula (Ia) is at least 20%.

4. 10. A compound of formula (I) according to any preceding claim, which is a mixture of (Ia) and (Ib), a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, wherein the enantiomeric excess (ee) of the enantiomer of formula (Ia) is at least 50%.

5. 10. A compound of formula (I) according to any preceding claim, which is a mixture of (Ia) and (Ib), a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, wherein the enantiomeric excess (ee) of the enantiomer of formula (Ia) is at least 80%.

6. 10. A compound of formula (I) according to any preceding claim, which is a mixture of (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, wherein the enantiomeric excess (ee) of the enantiomer of formula (Ia) is at least 99%.

7. R 2 is hydrogen, or an enantiomeric mixture comprising compounds of formula (I) or (Ia) or (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, according to any of the preceding claims.

8. R 2 is F, or an enantiomeric mixture comprising compounds of formula (I) or (Ia) or (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 6.

9. Compounds of formula (A) and (B), 10. A compound according to any preceding claim, selected from compounds (A) and (B) and mixtures thereof with their enantiomers.

10. A compound of formula (I), (Ia), (Ib), an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9, for use as a medicament.

11. A compound of formula (I) or (Ia) or (Ib), or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9 for use in the treatment and / or prevention of disease.

12. A compound of formula (I) or (Ia) or (Ib), or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9, for use according to claim 11 in the treatment and / or prevention of a proliferative or genetic disorder.

13. A compound of formula (I) or (Ia) or (Ib), or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9, for use according to claim 12 in the treatment and / or prevention of a proliferative or genetic disease involving oncogenic RAS or eIF4A complexes which control the expression of genes selected from c-myc and cyclin D1.

14. 10. A compound of formula (I) or (Ia) or (Ib), or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9, wherein the disease is selected from tumor proliferative diseases, cancer, RASO diseases, and inflammatory diseases, in particular CFC (craniofacial skin disease) and NF1 (neurofibromatosis type 1).

15. A compound of formula (I) or (Ia) or (Ib), or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9 for use as an inhibitor of RAS protein activation.

16. A compound of formula (I) or (Ia) or (Ib), or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9, for use as an inhibitor or ligand of the eIF4A complex or as a ligand of prohibitin.

17. A compound of formula (I) or (Ia) or (Ib), or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9 for use in the treatment of a proliferative disorder in which RAS signalling is involved.

18. A compound of formula (I) or (Ia) or (Ib), or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9 for use in the treatment of a proliferative disorder involving eIF4A-Myc signaling.

19. 18. A compound of formula (I) or (Ia) or (Ib), or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9, for use according to claim 17 in the treatment of a proliferative disorder involving KRAS G12V, NRAS G12V, HRAS G12V, KRAS G12C, KRAS G12D, KRAS G12C / Y96D, KRAS G13C, KRAS G13D, KRASG13S, KRAS Q61H, KRAS Q61R, or KRAS Q61K.

20. A compound of formula (I) or (Ia) or (Ib), or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9, for use according to claim 18 in the treatment of proliferative disorders in which c-myc is overexpressed and / or cyclin D1 is overexpressed.

21. A compound of formula (I) or (Ia) or (Ib), or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9, for use according to claim 17 in the treatment of a proliferative disorder involving any activating mutation in KRAS, HRAS and NRAS.

22. A compound of formula (I) or (Ia) or (Ib), or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9, for use according to claim 17 in the treatment of a proliferative disorder involving any mutation conferring resistance to RAS inhibitors.

23. A compound of formula (I) or (Ia) or (Ib), or an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9, for use according to claim 17 in the treatment of a proliferative disease in which mutations in EGFR or its related genes contribute to acquired resistance to RAS inhibitors.

24. A method of inhibiting the growth, proliferation, or metastasis of cancer cells in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of at least one compound selected from the compounds of formula (I), (Ia), (Ib), an enantiomeric mixture comprising the compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9.

25. A pharmaceutical composition comprising a pharmaceutically effective amount of at least one compound selected from the group consisting of compounds of formula (I), (Ia), (Ib), an enantiomeric mixture comprising compounds of formula (Ia) and (Ib), or a prodrug or isotopically enriched form thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9, and one or more pharmaceutically acceptable carriers.

26. 26. The pharmaceutical composition according to claim 25 for use in the prevention and / or treatment of a proliferative or genetic disorder.

27. a1) at least one compound selected from the compounds of formula (I), (Ia), (Ib), enantiomeric mixtures comprising compounds of formula (Ia) and (Ib), or prodrugs or isotopically enriched forms thereof, or pharmaceutically acceptable salts thereof, as defined in any one of claims 1 to 9; or a2) The pharmaceutical composition according to claim 25 A formulation comprising: b) Instructions for administering said pharmaceutical composition for the treatment of a disease in which inhibition of RAS activation or downstream signaling pathways, in particular C-Myc upregulation and / or cyclin D1 upregulation, is effective in treating the disease. Includes a kit.

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