Macrocyclic RAS inhibitors

Abstract

This disclosure deals with macrocyclic compounds of formulas (Ia) and (Ib) that can inhibit the Ras protein, as well as pharmaceutical compositions and protein conjugates thereof, and their use in the treatment of cancer. JPEG2026515696000242.jpg54170

Description

[Background technology]

[0001] The vast majority of small molecule drugs act by binding to functionally important pockets on target proteins, thereby regulating the activity of those proteins. For example, cholesterol-lowering drugs known as statins bind to the enzymatic active site of HMG-CoA reductase, thus preventing the enzyme from engaging with its substrate. The fact that many such drug / target interaction pairs are known can mislead some into believing that, with a reasonable amount of time, effort, and resources, small molecule modulators can be discovered for most, if not all, proteins. This is far from the truth. Current estimates suggest that only about 10% of all human proteins are targetable by small molecules. Non-patent document 1. The other 90% are currently considered unmanageable or difficult to address with the small molecule drug discovery described above. Such targets are commonly referred to as "undruggable." These undruggable targets include a vast and largely untapped treasure trove of medically important human proteins. Therefore, there is considerable interest in discovering novel molecular modalities that can modulate the function of such undruggable targets.

[0002] The literature has well established that Ras proteins (K-Ras, H-Ras, and N-Ras) play essential roles in various human cancers and are therefore suitable targets for anticancer therapy. In fact, mutations in the Ras protein account for approximately 30% of all human cancers in the United States, many of which are lethal. Dysregulation of the Ras protein due to activating mutations, overexpression, or upstream activation is common in human tumors, and activating mutations in Ras are frequently observed in human cancers. For example, an activating mutation at codon 12 of the Ras protein functions by inhibiting both the GTPase-activated protein (GAP)-dependent and intrinsic hydrolysis rates of GTP, significantly distorting the population of Ras mutant proteins into an "on" (GTP-bound) state (Ras(ON)), leading to oncogenic MAPK signaling. In particular, Ras exhibits picomolar affinity for GTP, allowing Ras to be activated even in the presence of low concentrations of this nucleotide. Mutations in codon 13 of Ras (e.g., G13C) and codon 61 (e.g., Q61H or Q61K) also contribute to oncogenic activity in some cancers.

[0003] Despite extensive drug discovery efforts against Ras over the past few decades, only two drugs targeting the K-Ras G12C variant (sotrasib and adagrasib) have been approved in the United States. Further efforts are needed to identify additional drugs for cancers driven by various Ras mutations. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Bojadzic and Buchwald,Curr Top Med Chem 18:674-699(2019) [Overview of the project]

[0005] This specification provides Ras inhibitors. The approach described herein involves the formation of a high-affinity triplicate or conjugate between a synthetic ligand and two intracellular proteins that do not normally interact under physiological conditions: namely, the protein of interest (e.g., Ras) and a widely expressed cytoplasmic chaperone (presenter protein) (e.g., cyclophyllin A). More specifically, in some embodiments, the Ras inhibitors described herein induce a novel binding pocket in Ras by driving the formation of a high-affinity triplicate or conjugate between the Ras protein and cyclophyllin A (CYPA), a widely expressed cytoplasmic chaperone. While not theoretically bound, the inventors believe that one way in which the compounds of the present invention and the complexes or conjugates they form achieve inhibitory effects on Ras is through steric occlusion of the interaction site between Ras and downstream effector molecules such as RAF and PI3K, which are necessary for propagating oncogenic signals.

[0006] Therefore, in some embodiments, this disclosure relates to formula Ia or formula Ib:

[0007] [ka]

[0008] This describes a compound having the structure of, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof [in the formula, A is a 3-6 member heterocycloalkylene that can be optionally substituted, a 3-6 member cycloalkylene that can be optionally substituted, a 6 member arylene that can be optionally substituted, or a 5-10 member heteroarylene that can be optionally substituted. L is a linker, R 1 These are 5-10 member heteroaryls that are optionally substituted, R 2 These are C1-C6 alkyl groups that are optionally substituted. R3 is C1-C6 alkyl optionally substituted, C1-C3 heteroalkyl optionally substituted, or 3-6 member cycloalkyl optionally substituted, R 4 is hydrogen or C1-C6 alkyl optionally substituted, each R 33 is independently halogen, C1-C3 alkyl optionally substituted, C1-C3 alkoxy optionally substituted, 3-6 member cycloalkyl optionally substituted, or 3-6 member heterocycloalkyl optionally substituted, t is 0, 1, 2, or 3, z is 0, 1, or 2, X 9 is -NR L6 -, -C(O)-, or -S(O)2-, R L1 、R L2 、R L3 、R L4 、R L4 、R L5 、and each of R L6 is independently hydrogen, halogen, hydroxyl, C1-C6 alkyl optionally substituted, C2-C6 alkenyl optionally substituted, C2-C6 alkynyl optionally substituted, or C1-C6 heteroalkyl optionally substituted, or alternatively any two of R L1 、R L2 、R L3 、R L4 、R L4 、R L5 、and R L6 together with the atoms to which they are attached and any intervening atoms form a C3-C8 cycloalkyl or 3-8 member heterocyclyl optionally substituted.

[0009] In some embodiments, the disclosure provides structural formula Ia-2:

[0010]

Chemical formula

[0011] This formula deals with compounds of the same, or pharmaceutically acceptable salts, enantiomers, stereoisomers, or tautomers thereof [in the formula, A is a 3-6 member heterocycloalkylene that can be optionally substituted, a 3-6 member cycloalkylene that can be optionally substituted, a 6 member arylene that can be optionally substituted, or a 5-10 member heteroarylene that can be optionally substituted. L is a linker, R 1 These are 5-10 member heteroaryls that are optionally substituted, R 2 These are C1-C6 alkyl groups that are optionally substituted. R 3 These are optionally substituted C1-C6 alkyl groups, optionally substituted C1-C3 heteroalkyl groups, or optionally substituted 3-6 member cycloalkyl groups. R 4 [These are C1-C6 alkyl groups that are substituted with hydrogen or optionally.]

[0012] Furthermore, pharmaceutical compositions comprising a compound of formula I, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient are also provided. Additionally, pharmaceutical compositions comprising a compound selected from Tables 1 and 2, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient are also provided.

[0013] Furthermore, a method for treating cancer in a subject requiring cancer treatment is also provided, which comprises administering to the subject a therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof.

[0014] In some embodiments, a method is provided for treating a Ras protein-related disorder in a subject requiring treatment for the disorder, the method comprising administering to the subject a therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof.

[0015] Furthermore, a method for inhibiting intracellular Ras protein is provided, which comprises contacting cells with an effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof. Any limitations discussed in relation to one embodiment of the present invention are specifically intended to be applicable to any other embodiment of the present invention. Furthermore, any compound or composition of the present invention may be used in any manner of the present invention, and any compound or composition of the present invention may be produced or utilized using any manner of the present invention.

[0016] Definitions and Chemical Terms In this application, unless otherwise evident from the context, (i) the term “a” means “one or more”; (ii) the term “or” is used to mean “and / or” unless it is explicitly indicated that it refers only to the options or that such options are not mutually exclusive, but this disclosure supports the definitions of options only and “and / or”; (iii) the terms “comprising” and “including” are understood to encompass the subdivided components or steps, whether they are presented by themselves or together with one or more additional components or steps; and (iv) if a scope is provided, the endpoints are included.

[0017] As used herein, the term “approximately” is used to indicate that a value includes the standard deviation of errors of the device or method used to determine that value. In certain embodiments, unless otherwise specified or evident from the context (for example, if such a number exceeds 100% of a possible value), the term “approximately” refers to a range of values ​​that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction from a given value (greater than or less than that).

[0018] As used herein, the term “adjacent” in the context of describing adjacent atoms refers to divalent atoms that are directly linked by a covalent bond. As used herein, “compounds of the present invention” and similar terms refer to the Ras inhibitors described herein, including, but not limited to, compounds of formula I and its subformulas, e.g., the compounds of Table 1 or Table 2, as well as their salts (e.g., pharmaceutically acceptable salts), solvates, hydrates, stereoisomers (including atropisomers), and tautomers.

[0019] The term "wild-type" refers to an entity that possesses a "normal" state or structure or activity similar to that found in nature (in contrast to mutant, pathological, altered, etc.). Those skilled in the art will understand that wild-type genes and polypeptides often exist in multiple different forms (e.g., alleles).

[0020] Those skilled in the art will understand that certain compounds described herein may exist in one or more different isomeric forms (e.g., stereoisomers, geometric isomers, atropisomers, tautomers) or isotopic forms (e.g., hydrogen substituted with deuterium, in which one or more atoms are substituted with different isotopes of that atom). Unless otherwise indicated or otherwise evident from the context, the illustrated structures can be understood to represent any such isomeric or isotopic forms individually or in combination.

[0021] The compounds described herein may be asymmetric (e.g., having one or more stereocenters). Unless otherwise indicated, all stereoisomers, such as enantiomers and diastereomers, are intended. Compounds of this disclosure containing asymmetrically substituted carbon atoms may be isolated in optically active or racemic forms. Methods for preparing optically active forms from optically active starting materials are known in the art, for example, by the separation of racemic mixtures or by stereoselective synthesis. Many geometric isomers, such as olefins and C=N double bonds, may also be present in the compounds described herein, and all such stable isomers are intended in this disclosure. Cis and trans geometric isomers of the compounds of this disclosure are described, and these may be isolated as mixtures of isomers or as separated isomers.

[0022] In some embodiments, one or more compounds illustrated herein may exist in different tautomerized forms. Unless explicitly excluded, references to such compounds encompass all such tautomerized forms, as will be apparent from the context. In some embodiments, the tautomerized form results from the exchange of a single bond with an adjacent double bond and the accompanying transfer of a proton. In certain embodiments, the tautomerized form may be a prototropic tautomer, which is an isomer protonated state having the same empirical formula and total charge as the reference form. Examples of moieties having a prototropic tautomerized form include ketone-enol pairs, amide-imido acid pairs, lactam-lactim pairs, amide-imido acid pairs, enamine-imine pairs, and cyclic forms in which protons may occupy two or more positions in the heterocyclic system, such as 1H- and 3H-imidazoles, 1H-, 2H-, and 4H-1,2,4-triazoles, 1H- and 2H-isoindoles, and 1H- and 2H-pyrazoles. In some embodiments, the tautomer morphs may be in equilibrium or sterically fixed to one morph by appropriate substitution. In certain embodiments, the tautomer morphs arise from acetal interconversion.

[0023] Unless otherwise indicated, the structures illustrated herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that may be incorporated into the compounds of the present invention include: 2 H, 3 H, 11 C, 13 C, 14 C, 13 N, 15 N, 15 , 17 O, 18 O, 32 P, 33 P, 35 S, 18 F, 36 Cl, 123 I and 125 This includes isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as I. Isotope-labeled compounds (e.g., 3 H and 14 (labeled with 1C) may be useful in compound or substrate tissue distribution assays. Tritiation (i.e., 3 H) and carbon-14 (i.e., 14 C) Isotopes may be useful due to their ease of preparation and detectability. Furthermore, deuterium (i.e., 2 Substitution with heavier isotopes such as H) may result in certain therapeutic benefits (e.g., increased in vivo half-life or reduced required dosage) due to greater metabolic stability. In some embodiments, one or more hydrogen atoms are replaced. 2 H or 3 Replaced by H, or one or more carbon atoms, 13 C- or 14 C is replaced by concentrated carbon. 15 O, 13 N, 11 C, and 18Positron-emitting isotopes such as fluorine are useful in positron emission tomography (PET) studies to investigate substrate receptor occupancy. The preparation of isotope-labeled compounds is known to those skilled in the art. For example, isotope-labeled compounds can generally be prepared by the following procedure, similar to the procedure disclosed for the compounds of the present invention described herein, by substituting an isotope-labeling reagent for a non-isotopically labeled reagent.

[0024] Non-limiting examples of moieties in the compound of the present invention that may contain one or more deuterium substitutions (where any position "R" can be deuterium (D)) include:

[0025] [ka]

[0026] These are some examples. Additional examples include:

[0027] [ka]

[0028] parts such as, and similar R 1 One example is the deuteration of the type portion [R 1 The definition of is found herein (for example, in compounds of formulas Ia, Ib, Ia-1, Ia-2, IIa, IIb, IIa-1, IIa-2, IIIa, IIIb, IIIa-1, IIIa-2, Va, Vb, Va-1, Va-2, VIIa, VIIb, VIIa-1 or VIIa-2, and their subformulas). Furthermore, the available positions in any A portion of the compounds of the formulas described herein, for example,

[0029] [ka]

[0030] Deuteration is also intended. Furthermore, deuterium substitution is also intended for the linker position in the compounds of the present invention, for example,

[0031] [ka]

[0032] It may also occur at the linker position, for example, in the compounds of the present invention.

[0033] [ka]

[0034] It may occur in [location]. Furthermore, deuterium substitution also affects the linker position in the compounds of the present invention, for example,

[0035] [ka]

[0036] It may occur in this way. In further embodiments, silylation substitutions, such as in the following linker, are also considered.

[0037] [ka]

[0038] As is known in the art, many chemical substances can exist in various different solid forms, such as amorphous or crystalline forms (e.g., polymorphs, hydrates, solvates). In some embodiments, the compounds of the present invention may be used in any such form, including any solid form. In some embodiments, the compounds described or illustrated herein may be provided or used in hydrate or solvate form.

[0039] In various parts of this specification, substituents of the compounds of this disclosure are disclosed by group or range. This disclosure is expressly intended to include all individual partial combinations of members of such groups and ranges. For example, the term "C1-C6 alkyl" is expressly intended to disclose methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl individually. Furthermore, where a compound includes multiple positions and substituents at those positions are disclosed by group or range, this disclosure is intended to include individual compounds and groups of compounds (e.g., genera and sub-genera) containing all individual partial combinations of members at each position, unless otherwise indicated.

[0040] The term “optionally substituted X” (e.g., “optionally substituted alkyl”) is intended to be equivalent to “X, where X is optionally substituted” (e.g., “alkyl, where the alkyl is optionally substituted”). It is not intended to mean that feature “X” (e.g., alkyl) itself is optional. When described herein, a particular compound of interest may contain one or more “optionally substituted” moieties. In general, the term “substituted,” whether preceded by the term “optionally,” means that one or more hydrogens of a designated moiety are replaced by preferred substituents, e.g., any substituents or groups described herein. Unless otherwise specified, an “optionally substituted” group may have preferred substituents at each of its substituted positions, and if more than one position in any given structure can be replaced by more than one substituent selected from the specified group, the substituents may be the same or different at all positions. For example, in the term "optionally substituted C1-C6 alkyl-C2-C9 heteroaryl," the alkyl moiety, the heteroaryl moiety, or both may be optionally substituted. The substituent combinations envisioned in this disclosure preferably result in the formation of stable or chemically feasible compounds. As used herein, the term "stable" means a compound that remains substantially unchanged when subjected to conditions that enable their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.

[0041] Suitable monovalent substituents on the replaceable carbon atoms of the "optionally substituted" groups are, independently, deuterium, halogen, -(CH2)0-4R°, -(CH2)0-4OR°, -O(CH2)0-4R°, -O-(CH2)0-4C(O)OR°, -(CH2)0-4CH(OR°)2, -(CH2)0-4SR°, -(CH2)0-4Ph (which may be substituted with R°), -(CH2)0-4O(CH2)0-1Ph (which may be substituted with R°), -CH=CHPh (which may be substituted with R°), -(CH2)0-4O(CH2)0-1-Ph Lysyl (which may be substituted with R°), 4- to 8-membered saturated or unsaturated heterocycloalkyl (e.g., pyridyl) (which may be substituted with R°), 3- to 8-membered saturated or unsaturated cycloalkyl (e.g., cyclopropyl, cyclobutyl, or cyclopentyl), -NO2, -CN, -N3, -(CH2)0-4N(R°)2, -(CH2)0-4N(R°)C(O)R°, -N(R°)C(S)R°, -(CH2)0-4N(R°)C(O)NR°2, -N(R°)C(S)NR°2, -(CH2)0-4N(R°)C(O)OR°, - N(R°)N(R°)C(O)R°, -N(R°)N(R°)C(O)NR°2, -N(R°)N(R°)C(O)OR°, -(CH2) 0-4C(O)R°, -C(O)R°, -C(S)R°, -(CH2)0-4C(O)OR°, -(CH2)0-4-C(O)-N(R°) 2, -(CH2)0-4-C(O)-N(R°)-S(O)2-R°, -C(NCN)NR°2, -(CH2)0-4C(O)SR°, -(CH2)0-4C(O)OSiR°3, -(CH2)0-4OC(O)R°, -OC(O)(CH2)0-4SR°, -SC(S)SR° , -(CH2)0-4SC(O)R°, -(CH2)0-4C(O)NR°2, -C(S)NR°2, -C(S)SR°, -(CH2)0-4OC(O)NR°2, -C(O)N(OR°)R°, -C(O)C(O)R°, -C(O)CH2C(O)R°, -C(NOR°)R° , -(CH2)0-4SSR°, -(CH2)0-4S(O)2R°, -(CH2)0-4S(O)2OR°, -(CH2)0-4OS(O)2R°, -S(O)2NR°2, -(CH2)0-4S(O)R°, -N(R°)S(O)2NR°2, -N(R°)S(O)2R°,-N(OR°)R°, -C(NOR°)NR°2, -C(NH)NR°2, -P(O)2R°, -P(O)R°2, -P(O)(OR°)2, -OP(O)R°2, -OP(O)(OR°)2, -OP(O)(OR°)R°, -SiR°3, -(C1-4 linear or branched alkylene)ON(R°)2, or -(C1-4 linear or branched alkylene)C(O)ON(R°)2, where each R° may be substituted as defined below, independently of hydrogen, -C1-6 aliphatic, -CH2Ph, -O(CH2)0-1Ph, -CH2-(5-6 membered heteroaryl ring), or 3-8 membered saturated or R° is an unsaturated cycloalkyl (e.g., cyclopropyl, cyclobutyl, or cyclopentyl), or a 3-6 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the above definition, two independent occurrences of R° together with their intervening atom(s) to form a 3-12 member monocyclic or bicyclic saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which may be substituted as defined below.

[0042] A suitable monovalent substituent on R° (or the ring formed by two independent occurrences of R° together with their intercalating atoms) is, independently, a halogen, -(CH2)0-2R ● ,-(HaroR ● ), -(CH2)O-2OH, -(CH2)O-2OR ● ,-(CH2)0-2CH(OR ● )2, -O(HaroR ● ), -CN, -N3, -(CH2)0-2C(O)R ● , -(CH2)0-2C(O)OH, -(CH2)0-2C(O)OR ● ,-(CH2)0-2SR ● , -(CH2)0-2SH, -(CH2)0-2NH2, -(CH2)0-2NHR ● ,-(CH2)0-2NR ● 2, -NO2, -SiR ● 3. -OSiR● 3. -C(O)SR ● 、 -(C1-4 linear or branched alkylene)C(O)OR ● , or -SSR ● This is possible, and in the formula, each R ● R° is either unsubstituted or, if preceded by "halo", substituted with only one or more halogens, and is independently selected from C1-4 aliphatic, -CH2Ph, -O(CH2)0-1Ph, or a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Preferred divalent substituents on the saturated carbon atom of R° include =O and =S.

[0043] Suitable divalent substituents on the saturated carbon atom of the "optionally substituted" group include, namely, =O, =S, =NNR * 2. =NNHC(O)R * 、=NNHC(O)OR * ,=NNHS(O)2R * ,=NR * 、=NOR * , -O(C(R * 2))2-3O-, or -S(C(R * 2))2-3S- is included, and in the formula, R * Each independent occurrence is selected from unsubstituted 5-6 membered saturated, partially unsaturated, or aryl rings having 0-4 heteroatoms independently selected from hydrogen, C1-6 aliphatic (which may be substituted as defined below), or nitrogen, oxygen, or sulfur. A preferred divalent substituent bonded to the vicinal, substituted carbon of the "optionally substituted" group is -O(CR * 2) Contains 2-3O-, in the formula, R * Each independent occurrence is selected from unsubstituted 5-6 member saturated, partially unsaturated, or aryl rings having 0-4 heteroatoms independently selected from hydrogen, C1-6 aliphatic atoms (which may be substituted as defined below), or nitrogen, oxygen, or sulfur.

[0044] R* Suitable substituents on the aliphatic group include halogen, -R ● , -(haloR ● ), -OH, -OR ● , -O(haloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2, or -NO2, wherein each R ● is unsubstituted or, when preceded by "halo", is substituted only with one or more halogens and is independently C1-4 aliphatic, -CH2Ph, -O(CH2)0-1Ph, or a 5-6 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0045] Suitable substituents on the nitrogen of a "optionally substituted" group include -R † , -NR † 2, -C(O)R † , -C(O)OR † , -C(O)C(O)R † , -C(O)CH2C(O)R † , -S(O)2R † , -S(O)2NR † 2, -C(S)NR † 2, -C(NH)NR † 2, or -N(R<00001​​​​​​​

[0046] R † Suitable substituents on the aliphatic group are, independently, halogens, -R ● ,-(HaroR ● ), -OH, -OR ● ,-O(HaroR ● ), -CN, -C(O)OH, -C(O)OR ● -NH2, -NHR ● , -NR ● 2, or -NO2, where each R ● R is either unsubstituted, or if preceded by "halo", substituted with only one or more halogens, and independently is a 5-6 member saturated ring, partially unsaturated ring, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or C1-4 aliphatic, -CH2Ph, -O(CH2)0-1Ph. † Suitable divalent substituents on the saturated carbon atom include =O and =S.

[0047] As used herein, the term "acetyl" refers to the group -C(O)CH3. As used herein, the term "alkoxy" refers to a compound in which an alkoxy group is bonded to the remainder of the compound via an oxygen atom, -O-C1~C 20 This refers to an alkyl group.

[0048] As used herein, the term “alkyl” refers to a saturated, linear or branched monovalent hydrocarbon group containing 1 to 20 (e.g., 1 to 10 or 1 to 6) carbon atoms. In some embodiments, the alkyl group is unbranched (i.e., linear), and in some embodiments, the alkyl group is branched. Alkyl groups are exemplified by, but are not limited to, methyl, ethyl, n-propyl and isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl, and neopentyl.

[0049] As used herein, the term "alkylene" refers to a saturated divalent hydrocarbon group derived from a straight-chain or branched-chain saturated hydrocarbon by removing two hydrogen atoms, exemplified by methylene, ethylene, isopropylene, etc. x ~C y The term "alkylene" refers to an alkylene group having x to y carbon atoms. Exemplary values ​​for x are 1, 2, 3, 4, 5, and 6, and exemplary values ​​for y are 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, or 20 (e.g., C1-C6, C1-C 10 , C2~C 20 , C2~C6, C2~C 10 , or C2~C 20 (Alkylene). In some embodiments, the alkylene may be further substituted with one, two, three, or four substituents as defined herein.

[0050] As used herein, the term “alkenyl” refers to a monovalent linear or branched group containing one or more carbon-carbon double bonds, unless otherwise specified, comprising 2 to 20 carbon atoms (e.g., 2 to 6 or 2 to 10 carbon atoms), exemplified by ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, and 2-butenyl. Alkenyls include both cis and trans isomers. As used herein, the term “alkenylene” refers to a divalent linear or branched group containing one or more carbon-carbon double bonds, unless otherwise specified, comprising 2 to 20 carbon atoms (e.g., 2 to 6 or 2 to 10 carbon atoms).

[0051] As used herein, the term "alkynyl" refers to a monovalent linear or branched group of 2 to 20 carbon atoms (e.g., 2 to 4, 2 to 6, or 2 to 10 carbons) containing a carbon-carbon triple bond, exemplified by ethynyl and 1-propynyl.

[0052] As used herein, the term "alkynyl sulfone" refers to a structure

[0053] [ka]

[0054] The formula represents a group containing, where R is any chemically feasible substituent as described herein. As used herein, the term "amino" refers to -N(R † )2, for example, represents -NH2 and -N(CH3)2.

[0055] As used herein, the term "aminoalkyl" refers to an alkyl moiety in which one or more carbon atoms are substituted with one or more amino moieties. As used herein, the term “amino acid” refers to a molecule having a side chain, an amino group, and an acid group (e.g., -CO2H or -SO3H), where the amino acid is bonded to a parent molecule's base by a side chain, an amino group, or an acid group (e.g., a side chain). As used herein, the term “amino acid” in its broadest sense refers to any compound or substance that can be incorporated into a polypeptide chain, for example, through the formation of one or more peptide bonds. In some embodiments, the amino acid has the general structure H2N-C(H)(R)-COOH. In some embodiments, the amino acid is a native amino acid. In some embodiments, the amino acid is a synthetic amino acid, in some embodiments, the amino acid is a D-amino acid, and in some embodiments, the amino acid is an L-amino acid. “Standard amino acid” refers to any of the 20 standard L-amino acids commonly found in native peptides. Exemplary amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, optionally substituted hydroxylnorvaline, isoleucine, leucine, lysine, methionine, norvaline, ornithine, phenylalanine, proline, pyrrolidine, selenocysteine, serine, taurine, threonine, tryptophan, tyrosine, and valine.

[0056] As used herein, the term "aryl" refers to a monovalent monocyclic, bicyclic, or polycyclic ring system formed by carbon atoms, wherein the ring bonded to the pendant group is aromatic. Examples of aryl groups include phenyl, naphthyl, phenantrenyl, and anthracenyl. The aryl ring can be bonded to its pendant group by any heteroatom or carbocyclic atom, thereby resulting in a stable structure, and unless otherwise specified, any of the ring atoms may be optionally substituted.

[0057] As used herein, the term "C0" represents a bond. For example, part of the term -N(C(O)-(C0~C5alkylene-H)- includes -N(C(O)-(C0alkylene-H)-, which is also represented by -N(C(O)-H)-.

[0058] As used herein, the terms "carbocyclic" and "carbocyclyl" refer to C3-C3 compounds that can be substituted with any monovalent compound. 12 This refers to monocyclic, bicyclic, or tricyclic ring structures, which may be bridged, condensed, or spirocyclic, where all rings are formed by carbon atoms and at least one ring is non-aromatic. Carbocyclic structures include cycloalkyl, cycloalkenyl, and cycloalkynyl groups. Examples of carbocyclyl groups include cyclohexyl, cyclohexenyl, cyclooctinyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, fluorenyl, indenyl, indanyl, and dekalinyl. Carbocyclic rings can be bonded to their pendant groups at any ring atom, thereby resulting in a stable structure, and unless otherwise specified, any of the ring atoms can be optionally substituted.

[0059] As used herein, the term "carbonyl" refers to a C(O) group, which can also be represented as C=O. As used herein, the term "carboxyl" means -CO2H, (C=O)(OH), COOH, or C(O)OH, or their unprotonated counterparts.

[0060] As used herein, the term "cyano" refers to the -CN group. As used herein, the term "cycloalkyl" refers to a monovalent saturated cyclic hydrocarbon group having 3 to 8 ring carbons, unless otherwise specified, which may be crosslinked, condensed, or spirocyclic, and is exemplified by cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cycloheptyl.

[0061] As used herein, the term "cycloalkenyl" refers to a monovalent, non-aromatic, saturated cyclic hydrocarbon group having 3 to 8 ring carbons and containing one or more carbon-carbon double bonds, which may be bridged, condensed, or spirocyclic.

[0062] As used herein, the term “diastereomer” refers to stereoisomers that cannot be superimposed on each other, rather than being mirror images of each other. As used herein, the term “enantiomer” means each individual optically active form of the compound of the present invention having an optical purity or enantiomer excess of at least 80% (i.e., at least 90% for one enantiomer and at most 10% for the other enantiomer), preferably at least 90%, and more preferably at least 98% (determined by methods standard in the art).

[0063] The term "guanidinyl" refers to the structure:

[0064] [ka]

[0065] This refers to a group having R, where each R is independently any chemically feasible substituent as described herein. As used herein, the term "guanidinoalkylalkyl" refers to an alkyl moiety in which one or more carbon atoms are substituted with one or more guanidinyl moieties.

[0066] As used herein, the term "haloacetyl" refers to an acetyl group in which at least one of its hydrogen atoms is replaced by a halogen. As used herein, the term "haloalkyl" refers to an alkyl moiety in which one or more carbon atoms are substituted by one or more identical or different halogen moieties.

[0067] As used herein, the term "halogen" refers to a halogen selected from bromine, chlorine, iodine, or fluorine. As used herein, the term “heteroalkyl” refers to an alkyl group as defined herein, in which at least one carbon atom is replaced by a heteroatom (e.g., an O, N, or S atom). The heteroatom may appear at the center or terminal of the radical.

[0068] As used herein, the term “heteroaryl” refers to a monovalent monocyclic or polycyclic ring structure containing at least one complete aromatic ring; that is, they contain 4n+2 pi electrons within the monocyclic or polycyclic ring system and contain at least one ring heteroatom selected from N, O, or S in the aromatic ring. Exemplary unsubstituted heteroaryl groups are those with 1 to 12 (e.g., 1 to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10, or 2 to 9) carbon atoms. The term “heteroaryl” also includes bicyclic, tricyclic, and tetracyclic groups in which any of the above heteroaromatic rings is fused to one or more aryl or carbocyclic rings, e.g., a phenyl ring or a cyclohexane ring. Examples of heteroaryl groups include, but are not limited to, pyridyl, pyrazolyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, imidazolyl, thiazolyl, quinolinyl, tetrahydroquinolinyl, and 4-azaindylol. The heteroaryl ring can be bonded to its pendant group at any ring atom, thereby resulting in a stable structure, and unless otherwise specified, any of the ring atoms can be optionally substituted. In some embodiments, the heteroaryl is substituted with 1, 2, 3, or 4 substituents.

[0069] As used herein, the term “heterocycloalkyl” refers to a monovalent monocyclic, bicyclic, or polycyclic ring system, which may be bridged, condensed, or spirocyclic, in which at least one ring is non-aromatic, and the non-aromatic ring contains 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. Five-membered rings have 0 to 2 double bonds, and six-membered and seven-membered rings have 0 to 3 double bonds. Exemplary unsubstituted heterocycloalkyl groups have 1 to 12 (e.g., 1 to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10, or 2 to 9) carbon atoms. The term “heterocycloalkyl” also refers to heterocyclic compounds having a bridged polycyclic structure in which one or more carbon or heteroatoms bridge two non-adjacent members of a monocyclic ring (e.g., quinuclidinyl groups). The term "heterocycloalkyl" includes bicyclic, tricyclic, and tetracyclic groups in which any of the above heterocyclic rings is fused to one or more aromatic rings, carbocyclic rings, heteroaromatic rings, or heterocyclic rings, such as an aryl ring, cyclohexane ring, cyclohexene ring, cyclopentane ring, cyclopentene ring, pyridine ring, or pyrrolidine ring. Examples of heterocycloalkyl groups are pyrrolidinyl, piperidinyl, 1,2,3,4-tetrahydroquinolinyl, decahydroquinolinyl, dihydropyrrolopyridine, and decahydronaphthilidinyl. Heterocycloalkyl rings can be bonded to their pendant group at any ring atom, thereby resulting in a stable structure, and any of the ring atoms may be optionally substituted unless otherwise specified.

[0070] As used herein, the term "hydroxy" refers to the -OH group. As used herein, the term "hydroxyalkyl" refers to an alkyl moiety in which one or more carbon atoms are substituted with one or more -OH moieties.

[0071] As used herein, the term “isomer” means any tautomer, stereoisomer, atropisomer, enantiomer, or diastereomer of any compound of the present invention. Compounds of the present invention may have one or more chiral centers or double bonds and are therefore recognized to exist as stereoisomers, e.g., double bond isomers (i.e., geometric E / Z isomers), or diastereomers (i.e., enantiomers (i.e., (+) or (-)), or cis / trans isomers). According to the present invention, the chemical structures illustrated herein, and therefore the compounds of the present invention, encompass both all corresponding stereoisomers, i.e., stereoisomerically pure forms (e.g., geometrically pure, enantiomerically pure, or diastereoisomerically pure) and mixtures of enantiomers and stereoisomers, e.g., racemates. Mixtures of enantiomers and stereoisomers of the compounds of the present invention can typically be separated into their constituent enantiomers or stereoisomers by known methods, such as chiral phase gas chromatography, chiral phase high-performance liquid chromatography, crystallization of the compound as a chiral salt complex, or crystallization of the compound in a chiral solvent. Enantiomers and stereoisomers can also be obtained from stereoisomerically or enantiomerically pure intermediates, reagents, and catalysts by known asymmetric synthesis methods.

[0072] As used herein, the term “linker” refers to a divalent organic moiety that links a first moiety (e.g., one moiety of a macrocyclic compound) to a second moiety (e.g., a second moiety of the same macrocyclic compound). In some embodiments, the linker results in compounds that can achieve an IC50 of 2 μM or less in the Ras-RAF interference assay protocol provided in the following examples and herein.

[0073] The purpose of this biochemical assay is to measure the ability of a test compound to promote the formation of a ternary complex between nucleotide-loaded Ras isoforms and cyclophyllin A. The resulting ternary complex is BRAF. RBDIt inhibits Ras signaling via RAF effectors by interfering with binding to the construct.

[0074] In an assay buffer containing 25 mM HEPES (pH 7.3), 0.002% Tween 20, 0.1% BSA, 100 mM NaCl, and 5 mM MgCl2, untagged cyclophyllin A, His6-K-Ras-GMPPNP (or other Ras variants), and GST-BRAF were added. RBD The compounds were combined in a 384-well assay plate at final concentrations of 25 μM, 12.5 nM, and 50 nM, respectively. The compounds were present in the plate wells as a 10-point 3-fold dilution series starting from a final concentration of 30 μM. After incubation at 25°C for 3 hours, the mixture of anti-His Eu-W1024 and anti-GST allophycocyanin was then added to the assay sample wells at final concentrations of 10 nM and 50 nM, respectively, and the reaction was incubated for a further 1.5 hours. The TR-FRET signal was read using a microplate reader (Ex 320 nm, Em 665 / 615 nm). Compounds that promote interference with the Ras:RAF complex were identified as those that induced a decrease in the TR-FRET ratio compared to the DMSO control well.

[0075] Selectivity may also be evaluated using this assay. In some embodiments, the compounds of the present invention are more selective to one or more specific Ras variants (e.g., K-Ras Q61H) than other Ras variants or wild types compared to those known in the art.

[0076] In some embodiments, the linker contains 20 or fewer linear atoms. In some embodiments, the linker contains 15 or fewer linear atoms. In some embodiments, the linker contains 10 or fewer linear atoms. In some embodiments, the linker has a molecular weight of less than 500 g / mol. In some embodiments, the linker has a molecular weight of less than 400 g / mol. In some embodiments, the linker has a molecular weight of less than 300 g / mol. In some embodiments, the linker has a molecular weight of less than 200 g / mol. In some embodiments, the linker has a molecular weight of less than 100 g / mol. In some embodiments, the linker has a molecular weight of less than 50 g / mol.

[0077] When used herein, the "monovalent organic moiety" is less than 500 kDa. In some embodiments, the "monovalent organic moiety" is less than 400 kDa. In some embodiments, the "monovalent organic moiety" is less than 300 kDa. In some embodiments, the "monovalent organic moiety" is less than 200 kDa. In some embodiments, the "monovalent organic moiety" is less than 100 kDa. In some embodiments, the "monovalent organic moiety" is less than 50 kDa. In some embodiments, the "monovalent organic moiety" is less than 25 kDa. In some embodiments, the "monovalent organic moiety" is less than 20 kDa. In some embodiments, the "monovalent organic moiety" is less than 15 kDa. In some embodiments, the "monovalent organic moiety" is less than 10 kDa. In some embodiments, the "monovalent organic moiety" is less than 1 kDa. In some embodiments, the "monovalent organic moiety" is less than 500 g / mol. In some embodiments, the "monovalent organic moiety" is in the range of 500 g / mol to 500 kDa.

[0078] As used herein, the term “stereoisomer” refers to all possible different isomeric and conformational forms that a compound (for example, a compound of any formula described herein) may have, in particular all possible stereochemical and stereostructural isomers of the basic molecular structure, including atropisomers, all diastereomers, enantiomers, or conformational isomers. Some of the compounds of the present invention may exist in different tautomeric forms, all of which are within the scope of the present invention.

[0079] As used herein, the term "sulfonyl" refers to the -S(O)2- group. As used herein, the term "thiocarbonyl" refers to the -C(S)- group. As used herein, the term "vinyl ketone" refers to a group containing a carbonyl group directly attached to a carbon-carbon double bond.

[0080] As used herein, the term "vinyl sulfone" refers to a group containing a sulfonyl group directly attached to a carbon-carbon double bond. As used herein, the term "ynone" refers to a structure.

[0081] [ka]

[0082] This refers to a group containing, where R is any chemically feasible substituent as described herein. Those skilled in the art will read this disclosure and understand that certain compounds described herein may be provided or used in any of various forms, such as salt forms, protected forms, prodrug forms, ester forms, isomeric forms (e.g., optical or structural isomers), isotopic forms, etc. In some embodiments, reference to a particular compound may refer to a particular form of that compound. In some embodiments, reference to a particular compound may refer to the compound in any form. In some embodiments, for example, a preparation of a single stereoisomer of a compound may be considered a different form of the compound from a racemic mixture of the compound; a particular salt of a compound may be considered a different form from another salt form of the compound; a preparation containing one conformational isomer of a double bond ((Z) or (E)) may be considered a different form from one containing the other conformational isomer of the double bond ((E) or (Z)); and a preparation in which one or more atoms are isotopes different from those present in the reference preparation may be considered a different form. [Modes for carrying out the invention]

[0083] compound This specification provides Ras inhibitors. The approach described herein involves the formation of a high-affinity triplicate or conjugate between a synthetic ligand and two intracellular proteins that do not normally interact under normal physiological conditions: namely, the protein of interest (e.g., Ras) and a widely expressed cytoplasmic chaperone (presenter protein) (e.g., cyclophyllin A). More specifically, in some embodiments, the Ras inhibitors described herein induce a novel binding pocket in Ras by driving the formation of a high-affinity triplicate or conjugate between the Ras protein and cyclophyllin A (CYPA), a widely expressed cytoplasmic chaperone. While not theoretically bound, the inventors believe that one way in which the compounds of the present invention and the complexes or conjugates they form achieve inhibitory effects on Ras is through steric occlusion of the interaction site between Ras and downstream effector molecules such as RAF, which are necessary for propagating oncogenic signals.

[0084] While not bound by theory, the inventors infer that non-covalent interactions between the compounds of the present invention and Ras and chaperone proteins (e.g., cyclophyllin A) may contribute to the inhibition of Ras activity. For example, van der Waals interactions, hydrophobic interactions, hydrophilic interactions, and hydrogen bonding interactions, as well as combinations thereof, may contribute to the ability of the compounds of the present invention to form complexes and act as Ras inhibitors.

[0085] Therefore, various Ras proteins can be inhibited by the compounds of the present invention (e.g., K-Ras, N-Ras, H-Ras, and their variants at positions 12, 13, and 61, e.g., G12C, G12D, G12V, G12S, G13C, G13D, Q61H, Q61K, Q61R, and Q61L, as well as others described herein, or combinations thereof). In some embodiments, the compounds of the present invention inhibit at least the K-Ras Q61H variant. In some embodiments, the compounds of the present invention selectively inhibit at least the K-Ras Q61H variant compared to wild-type K-Ras. In some embodiments, the compounds of the present invention selectively inhibit the K-Ras Q61H variant compared to wild-type K-Ras and one or more other K-Ras variants.

[0086] In some embodiments, this disclosure refers to formula Ia or formula Ib:

[0087] [ka]

[0088] This describes a compound having the structure of, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof [in the formula, A is a 3-6 member heterocycloalkylene that can be optionally substituted, a 3-6 member cycloalkylene that can be optionally substituted, a 6 member arylene that can be optionally substituted, or a 5-10 member heteroarylene that can be optionally substituted. L is a linker, R 1 These are 5-10 member heteroaryls that are optionally substituted, R 2 These are C1-C6 alkyl groups that are optionally substituted. R 3 These are optionally substituted C1-C6 alkyl groups, optionally substituted C1-C3 heteroalkyl groups, or optionally substituted 3-6 member cycloalkyl groups. R 4is a C1-C6 alkyl group that is substituted with hydrogen or optionally. Each R 33 These are independently halogens, optionally substituted C1-C3 alkyls, optionally substituted C1-C3 alkoxys, optionally substituted 3-6 membered cycloalkyls, or optionally substituted 3-6 membered heterocycloalkyls. t is 0, 1, 2, or 3. z is 0, 1, or 2. X 9 -NR L6 -, -C(O)-, or -S(O)2-, R L1 , R L2 , R L3 , R L4 , R L4 , R L5 , and R L6 Each of them independently is hydrogen, halogen, hydroxyl, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, or optionally substituted C1-C6 heteroalkyl, or R L1 , R L2 , R L3 , R L4 , R L4 , R L5 , and R L6 Any two of these, together with the atoms bonded to them and any intervening atoms, form a C3-C8 cycloalkyl or 3-8 membered heterocycline which can be optionally substituted.

[0089] In some embodiments, this disclosure relates to structural formula Ia-1:

[0090] [ka]

[0091] This formula deals with compounds of the same, or pharmaceutically acceptable salts, enantiomers, stereoisomers, or tautomers thereof [in the formula, A is a 3-6 member heterocycloalkylene that can be optionally substituted, a 3-6 member cycloalkylene that can be optionally substituted, a 6 member arylene that can be optionally substituted, or a 5-10 member heteroarylene that can be optionally substituted. L is a linker, R 1 These are 5-10 member heteroaryls that are optionally substituted, R 2 These are C1-C6 alkyl groups that are optionally substituted. R 3 These are optionally substituted C1-C6 alkyl groups, optionally substituted C1-C3 heteroalkyl groups, or optionally substituted 3-6 member cycloalkyl groups. R 4 is a C1-C6 alkyl group that is substituted with hydrogen or optionally. Each R 33 These are independently halogens, optionally substituted C1-C3 alkyls, optionally substituted C1-C3 alkoxys, optionally substituted 3-6 membered cycloalkyls, or optionally substituted 3-6 membered heterocycloalkyls. t is 0, 1, 2, or 3.

[0092] In one embodiment, the present invention relates to formula Ia-2:

[0093] [ka]

[0094] This describes a compound having the structure of, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof [in the formula, A is a 3-6 member heterocycloalkylene that can be optionally substituted, a 3-6 member cycloalkylene that can be optionally substituted, a 6 member arylene that can be optionally substituted, or a 5-10 member heteroarylene that can be optionally substituted. L is a linker, R 1 These are 5-10 member heteroaryls that are optionally substituted, R 2 These are C1-C6 alkyl groups that are optionally substituted. R 3 These are optionally substituted C1-C6 alkyl groups, optionally substituted C1-C3 heteroalkyl groups, or optionally substituted 3-6 member cycloalkyl groups. R 4 [These are C1-C6 alkyl groups that are substituted with hydrogen or optionally.]

[0095] In some embodiments, the compound is of formula IIa-2:

[0096] [ka]

[0097] It has the structure of, or is a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof [in the formula, R 5 This includes hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkenyl, optionally substituted C1-C6 alkynyl, optionally substituted 3-10 member heterocycloalkyl, and -OR 5a , or a C1-C6 heteroalkyl that is optionally substituted, R 5a [These are optionally substituted C1-C6 alkyl groups or optionally substituted 5-10 member heteroaryl groups.]

[0098] In some embodiments, R 5 This is a hydrogen atom, optionally substituted with a 3-10 member heterocycloalkyl group, or -OR 5a or optionally substituted C1-C6 heteroalkyl groups.

[0099] In some embodiments, the present disclosure relates to structural formula IIa or formula IIb:

[0100] [ka]

[0101] This formula deals with compounds of the same, or pharmaceutically acceptable salts, enantiomers, stereoisomers, or tautomers thereof [in the formula, R 5 This is a hydrogen atom, optionally substituted with a 3-10 member heterocycloalkyl group, or -OR 5a , or a C1-C6 heteroalkyl that is optionally substituted, R 5a [These are optionally substituted C1-C6 alkyl groups or optionally substituted 5-10 member heteroaryl groups.]

[0102] In some embodiments, the present disclosure relates to structural formula IIa-1:

[0103] [ka]

[0104] This formula deals with compounds of the same, or pharmaceutically acceptable salts, enantiomers, stereoisomers, or tautomers thereof [in the formula, R 5 This is a hydrogen atom, optionally substituted with a 3-10 member heterocycloalkyl group, or -OR 5a , or a C1-C6 heteroalkyl that is optionally substituted, R 5a These are optionally substituted C1-C6 alkyl groups or optionally substituted 5-10 member heteroaryl groups. Each R 33 These are independently halogens, optionally substituted C1-C3 alkyls, optionally substituted C1-C3 alkoxys, optionally substituted 3-6 membered cycloalkyls, or optionally substituted 3-6 membered heterocycloalkyls. t is 0, 1, 2, or 3.

[0105] In some embodiments, the present disclosure relates to structural formula IIa-2:

[0106] [ka]

[0107] This formula deals with compounds of the same, or pharmaceutically acceptable salts, enantiomers, stereoisomers, or tautomers thereof [in the formula, R 5 This is a hydrogen atom, optionally substituted with a 3-10 member heterocycloalkyl group, or -OR 5a , or a C1-C6 heteroalkyl that is optionally substituted, R 5a [These are optionally substituted C1-C6 alkyl groups or optionally substituted 5-10 member heteroaryl groups.]

[0108] In some embodiments, the present disclosure relates to structural formula IIIa-1:

[0109] [ka]

[0110] We deal with compounds of the same species, or their pharmaceutically acceptable salts, enantiomers, stereoisomers, or tautomers. In some embodiments, the present disclosure relates to structural formula IIIa-2:

[0111] [ka]

[0112] We deal with compounds of the same species, or their pharmaceutically acceptable salts, enantiomers, stereoisomers, or tautomers. In some embodiments, the compound is of formula II1a-1:

[0113] [ka]

[0114] It has the structure of, or is a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof. In some embodiments, L is given by Equation III:

[0115] [ka]

[0116] It has a structure, In the formula, X 1 It is either O or CH2 and is bonded to ring A. Z is a 3- to 6-membered heterocycloalkylene that can be optionally substituted, a C1- to C6 alkylene that can be optionally substituted, or a C1- to C6 heteroalkylene that can be optionally substituted.

[0117] In some embodiments, the compound is of formula Va-2:

[0118] [ka]

[0119] It has the structure of, or is a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof [in the formula, X 1 is either O or CH2, Z is a 3- to 6-membered heterocycloalkylene that is optionally substituted, a C1- to C6 alkylene that is optionally substituted, or a C1- to C6 heteroalkylene that is optionally substituted.

[0120] In some embodiments, X 1 It is O. In some embodiments, Z is a 3- to 6-membered heterocycloalkylene that is optionally substituted. In some embodiments, Z is a 5-membered heterocycloalkylene that is optionally substituted. In some embodiments, Z is a pyrrolidine-diyl that is optionally substituted. In some embodiments, Z is a pyrrolidine-diyl that is optionally substituted. In some embodiments, Z is a pyrrolidine-diyl that is optionally substituted.

[0121] In some embodiments, L is given by formula IV:

[0122] [ka]

[0123] It has a structure, X 1 It is either O or CH2 and is bonded to ring A. Z is a 3- to 6-membered heterocycloalkylene that can be optionally substituted, a C1- to C6 alkylene that can be optionally substituted, or a C1- to C6 heteroalkylene that can be optionally substituted.

[0124] In some embodiments, L is given by formula VI:

[0125] [ka]

[0126] It has a structure, In the formula, B is a 3- to 6-membered heterocycloalkylene that can be optionally substituted. R 6 This includes hydrogen, optionally substituted C1-C6 alkyl groups, optionally substituted C1-C6 heteroalkyl groups, optionally substituted 3-6 member heterocyclines, optionally substituted 3-6 member cycloalkyl groups, optionally substituted 5-10 member heteroaryl groups, and optionally substituted C6-C6 groups. 10 Ariel,

[0127] [ka]

[0128] And, R 7 and R 8 Each of these is independently a C1-C6 alkyl group that is substituted with H or optionally. R 9 These are optionally substituted C1-C6 alkyl groups, optionally substituted C2-C6 alkenyl groups, optionally substituted 3-6 membered cycloalkyl groups, or optionally substituted 3-6 membered heterocyclyl groups. R 10 This includes C1-C6 alkyl groups that can be optionally substituted, C2-C6 alkenyl groups that can be optionally substituted, C1-C6 heteroalkyl groups that can be optionally substituted, and C3-C6 groups that can be optionally substituted. 10 Cycloalkyl, optionally substituted 3-6 membered heterocyclyl, optionally substituted 5-10 membered heteroaryl, or optionally substituted C6-C 10 It is Ariel, R 11 This includes hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C2-C6 heteroalkynyl, and optionally substituted C3-C 10 Cycloalkyls, optionally substituted 3-10 member heterocyclines, optionally substituted C3-C 10 Cycloalkenyls, optionally substituted 3-10 member heterocycloalkenyls, optionally substituted C6-C 10 It is an aryl or a 5- to 10-membered heteroaryl that can be optionally substituted.

[0129] In some embodiments, L is given by formula VIa:

[0130] [ka]

[0131] It has the structure of [the object]. In some embodiments, the compound is of formula VIIa-2:

[0132] [ka]

[0133] It has the structure of, or is a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof [in the formula, R 6 This includes hydrogen, optionally substituted C1-C6 alkyl groups, optionally substituted C1-C6 heteroalkyl groups, optionally substituted 3-6 member heterocyclines, optionally substituted 3-6 member cycloalkyl groups, optionally substituted 5-10 member heteroaryl groups, and optionally substituted C6-C6 groups. 10 Ariel,

[0134] [ka]

[0135] And, R 7 and R 8 Each of these is independently a C1-C6 alkyl group that is substituted with H or optionally. R 9 These are optionally substituted C1-C6 alkyl groups, optionally substituted C2-C6 alkenyl groups, optionally substituted 3-6 membered cycloalkyl groups, or optionally substituted 3-6 membered heterocyclyl groups. R 10 This includes C1-C6 alkyl groups that can be optionally substituted, C2-C6 alkenyl groups that can be optionally substituted, C1-C6 heteroalkyl groups that can be optionally substituted, and C3-C6 groups that can be optionally substituted. 10 Cycloalkyl, optionally substituted 3-6 membered heterocyclyl, optionally substituted 5-10 membered heteroaryl, or optionally substituted C6-C 10It is Ariel, R 11 This includes hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C2-C6 heteroalkynyl, and optionally substituted C3-C 10 Cycloalkyls, optionally substituted 3-10 member heterocyclines, optionally substituted C3-C 10 Cycloalkenyls, optionally substituted 3-10 member heterocycloalkenyls, optionally substituted C6-C 10 [An aryl, or a 5- to 10-membered heteroaryl that is optionally substituted.]

[0136] In some embodiments, the compound is of formula VIIa or formula VIIb:

[0137] [ka]

[0138] It has the structure of, or is a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof [in the formula, R 6 This includes hydrogen, optionally substituted C1-C6 alkyl groups, optionally substituted C1-C6 heteroalkyl groups, optionally substituted 3-6 member heterocyclines, optionally substituted 3-6 member cycloalkyl groups, optionally substituted 5-10 member heteroaryl groups, and optionally substituted C6-C6 groups. 10 Ariel,

[0139] [ka]

[0140] And, R 7 and R 8Each of these is independently a C1-C6 alkyl group that is substituted with H or optionally. R 9 These are optionally substituted C1-C6 alkyl groups, optionally substituted C2-C6 alkenyl groups, optionally substituted 3-6 membered cycloalkyl groups, or optionally substituted 3-6 membered heterocyclyl groups. R 10 This includes C1-C6 alkyl groups that can be optionally substituted, C2-C6 alkenyl groups that can be optionally substituted, C1-C6 heteroalkyl groups that can be optionally substituted, and C3-C6 groups that can be optionally substituted. 10 Cycloalkyl, optionally substituted 3-6 membered heterocyclyl, optionally substituted 5-10 membered heteroaryl, or optionally substituted C6-C 10 It is Ariel, R 11 This includes hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C2-C6 heteroalkynyl, and optionally substituted C3-C 10 Cycloalkyls, optionally substituted 3-10 member heterocyclines, optionally substituted C3-C 10 Cycloalkenyls, optionally substituted 3-10 member heterocycloalkenyls, optionally substituted C6-C 10 [An aryl, or a 5- to 10-membered heteroaryl that is optionally substituted.]

[0141] In some embodiments, the compound is of formula VIIa-1:

[0142] [ka]

[0143] It has the structure of, or is a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof [in the formula, R 6This includes hydrogen, optionally substituted C1-C6 alkyl groups, optionally substituted C1-C6 heteroalkyl groups, optionally substituted 3-6 member heterocyclines, optionally substituted 3-6 member cycloalkyl groups, optionally substituted 5-10 member heteroaryl groups, and optionally substituted C6-C6 groups. 10 Ariel,

[0144] [ka]

[0145] And, R 7 and R 8 Each of these is independently a C1-C6 alkyl group that is substituted with H or optionally. R 9 These are optionally substituted C1-C6 alkyl groups, optionally substituted C2-C6 alkenyl groups, optionally substituted 3-6 membered cycloalkyl groups, or optionally substituted 3-6 membered heterocyclyl groups. R 10 This includes C1-C6 alkyl groups that can be optionally substituted, C2-C6 alkenyl groups that can be optionally substituted, C1-C6 heteroalkyl groups that can be optionally substituted, and C3-C6 groups that can be optionally substituted. 10 Cycloalkyl, optionally substituted 3-6 membered heterocyclyl, optionally substituted 5-10 membered heteroaryl, or optionally substituted C6-C 10 It is Ariel, R 11 This includes hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C2-C6 heteroalkynyl, and optionally substituted C3-C 10 Cycloalkyls, optionally substituted 3-10 member heterocyclines, optionally substituted C3-C 10 Cycloalkenyls, optionally substituted 3-10 member heterocycloalkenyls, optionally substituted C6-C10 [An aryl, or a 5- to 10-membered heteroaryl that is optionally substituted.]

[0146] In some embodiments, the compound is of formula VIIa-2:

[0147] [ka]

[0148] It has the structure of, or is a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof [in the formula, R 6 This includes hydrogen, optionally substituted C1-C6 alkyl groups, optionally substituted C1-C6 heteroalkyl groups, optionally substituted 3-6 member heterocyclines, optionally substituted 3-6 member cycloalkyl groups, optionally substituted 5-10 member heteroaryl groups, and optionally substituted C6-C6 groups. 10 Ariel,

[0149] [ka]

[0150] And, R 7 and R 8 Each of these is independently a C1-C6 alkyl group that is substituted with H or optionally. R 9 These are optionally substituted C1-C6 alkyl groups, optionally substituted C2-C6 alkenyl groups, optionally substituted 3-6 membered cycloalkyl groups, or optionally substituted 3-6 membered heterocyclyl groups. R 10 This includes C1-C6 alkyl groups that can be optionally substituted, C2-C6 alkenyl groups that can be optionally substituted, C1-C6 heteroalkyl groups that can be optionally substituted, and C3-C6 groups that can be optionally substituted. 10 Cycloalkyl, optionally substituted 3-6 membered heterocyclyl, optionally substituted 5-10 membered heteroaryl, or optionally substituted C6-C10 It is Ariel, R 11 This includes hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C2-C6 heteroalkynyl, and optionally substituted C3-C 10 Cycloalkyls, optionally substituted 3-10 member heterocyclines, optionally substituted C3-C 10 Cycloalkenyls, optionally substituted 3-10 member heterocycloalkenyls, optionally substituted C6-C 10 [An aryl, or a 5-10 member heteroaryl which can be optionally substituted.] In some embodiments, R 6 teeth,

[0151] [ka]

[0152] In some embodiments, R 11 R is a C1-C6 alkyl that is optionally substituted. In some embodiments, 11 R is a C2-C6 alkenyl that is optionally substituted. In some embodiments, R 11 R is a C2-C6 alkynyl that is optionally substituted. In some embodiments, R 11 is a C1-C6 heteroalkyl that is optionally substituted. In some embodiments, R 11 R is a C2-C6 heteroalkenyl that is optionally substituted. In some embodiments, R 11 R is a C2-C6 heteroalkynyl that is optionally substituted. In some embodiments, R 11 C3~C are replaced by optional selection. 10 It is a cycloalkenyl. In some embodiments, R 11 is hydrogen. In some embodiments, R 11 C3~C are replaced by optional selection.10 It is cycloalkyl. In some embodiments, R 11 These are 3- to 10-membered heterocyclines that are substituted by optional selection.

[0153] In some embodiments, R 6 teeth,

[0154] [ka]

[0155] In some embodiments, R 10 R is a 5-10 member heteroaryl that is optionally substituted. In some embodiments, R 10 These are 3- to 10-membered heterocyclines that are substituted by optional selection.

[0156] In some embodiments, R 6 These are 3- to 6-membered heterocyclines that are substituted by optional selection. In some embodiments, A is optionally substituted with a 3- to 6-membered heterocycloalkylene, optionally substituted with a 6-membered arylene, or optionally substituted with a 5- to 10-membered heteroarylene. In some embodiments, A is optionally substituted with a 6-membered arylene. In some embodiments, A is

[0157] [ka]

[0158] And in the formula,

[0159] [ka]

[0160] This represents the part of the molecule bonded to the linker. In some embodiments, A is

[0161] [ka]

[0162] That is the case. In some embodiments, R 2 is a C1-C3 alkyl or C1-C3 haloalkyl. In some embodiments, R 2 teeth,

[0163] [ka]

[0164] In some embodiments, R 2 teeth,

[0165] [ka]

[0166] That is the case. In some embodiments, R 4 R is a C1-C6 alkyl that is optionally substituted. In some embodiments, 4 It is methyl.

[0167] In some embodiments, R 3 R is a C1-C6 alkyl that is optionally substituted. In some embodiments, 3 R is a 3- to 6-membered cycloalkyl that is optionally substituted. In some embodiments, R 3 teeth,

[0168] [ka]

[0169] That is the case. In some embodiments, R 5 is hydrogen. In some embodiments, R 5This is a 3- to 10-membered heterocycloalkyl that can be optionally substituted.

[0170] In some embodiments, R 5 teeth,

[0171] [ka]

[0172] That is the case. In some embodiments, R 5 is -OR 5a In some embodiments, R 5 teeth,

[0173] [ka]

[0174] That is the case. In any embodiment of this specification, the compounds of the present invention may be modified with substituents found in one or more of the following applications, which are incorporated herein in whole by reference: WO2024 / 060966, WO2024 / 017859, WO2024 / 008834, WO2024 / 008610, WO2023 / 232776, WO2023 / 208005, WO2023 / 086341, WO2023 / 025832, WO2023 / 015559, CN117720556, CN117720555, CN117720554, CN117534687, CN117534685, and CN117534684.

[0175] In some embodiments, the compounds of the present invention are selected from Tables 1 and 2, or are pharmaceutically acceptable salts or stereoisomers thereof. In some embodiments, the compounds of the present invention are selected from Tables 1 and 2, or are pharmaceutically acceptable salts or atropisomers thereof.

[0176] [Table 1-1]

[0177] Table 1-2

[0178] Table 1-3

[0179] Table 1-4

[0180] Table 1-5

[0181] Table 1-6

[0182] Table 1-7

[0183] Table 1-8

[0184] Table 1-9

[0185] Table 1-10

[0186] Table 1-11

[0187] Table 1-12

[0188] Table 1-13

[0189] Table 1-14

[0190] Table 1-15

[0191] Table 1-16

[0192] Table 1-17

[0193] Table 1-18

[0194] Table 1-19

[0195] Table 1-20

[0196] Table 1-21

[0197] Table 1-22

[0198] Table 1-23

[0199] Table 1-24

[0200] Table 1-25

[0201] Table 1-26

[0202] Table 1-27

[0203] Table 1-28

[0204] Table 1-29

[0205] Table 1-30

[0206] Table 1-31

[0207] Table 1-32

[0208] Table 1-33

[0209] Table 1-34

[0210] Table 1-35

[0211] Table 1-36

[0212] Table 1-37

[0213] Table 1-38

[0214] Table 1-39

[0215] Table 1-40

[0216] Table 1-41

[0217] Table 1-42

[0218] Table 1-43

[0219] Table 1-44

[0220] Table 1-45

[0221] Table 1-46

[0222] Table 1-47

[0223] Table 1-48

[0224] Table 1-49

[0225] Table 1-50

[0226] Table 1-51

[0227] Table 1-52

[0228] Table 1-53

[0229] Table 1-54

[0230] Table 1-55

[0231] Table 2-1

[0232] Table 2-2

[0233] Table 2-3

[0234] Table 2-4

[0235] Table 2-5

[0236] Table 2-6

[0237] Furthermore, a pharmaceutical composition is also provided, comprising the compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. The compounds of the present invention are also suitable for use in antibody-drug conjugates and in applications as degrading agents.

[0238] Furthermore, methods for treating cancer in subjects requiring cancer treatment are also provided, the methods comprising administering to the subject a therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof. The cancer may be, for example, pancreatic cancer, colorectal cancer, non-small cell lung cancer, acute myeloid leukemia, multiple myeloma, thyroid cancer, myelodysplastic syndrome, or squamous cell carcinoma of the lung. In some embodiments, the cancer is pancreatic cancer, colorectal cancer, non-small cell lung cancer, acute myeloid leukemia, or multiple myeloma. In some embodiments, the cancer includes Ras mutations such as K-Ras Q61H, H-Ras Q61H, or N-Ras Q61H. Other Ras mutations are described herein.

[0239] Furthermore, a method is provided for treating Ras protein-related disorders in subjects requiring treatment of such disorders, the method comprising administering to the subject a therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof.

[0240] Furthermore, a method for inhibiting intracellular Ras proteins is provided, comprising contacting cells with an effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof. For example, the Ras protein is K-Ras Q61H, H-Ras Q61H, or N-Ras Q61H. Other Ras proteins are described herein. The cells may be cancer cells such as pancreatic cancer cells, colorectal cancer cells, non-small cell lung cancer cells, acute myeloid leukemia cells, multiple myeloma cells, thyroid cancer cells, myelodysplastic syndrome cells, or lung squamous cell carcinoma cells. In some embodiments, the cells are pancreatic cancer cells, colorectal cancer cells, non-small cell lung cancer cells, acute myeloid leukemia cells, or multiple myeloma cells. Other cancer types are described herein. The cells may be in vivo or in vitro.

[0241] With respect to the compounds of the present invention, one stereoisomer may exhibit better inhibition than another. For example, one atropisomer may exhibit inhibition, while the other atropisomer may exhibit little to no inhibition.

[0242] In some embodiments, the methods or uses described herein further include administering additional anticancer agents. In some embodiments, the additional anticancer agents are HER2 inhibitors, EGFR inhibitors, second Ras inhibitors, SHP2 inhibitors, SOS1 inhibitors, Raf inhibitors, MEK inhibitors, ERK inhibitors, PI3K inhibitors, PTEN inhibitors, AKT inhibitors, mTORC1 inhibitors, BRAF inhibitors, PD-L1 inhibitors, PD-1 inhibitors, CDK4 / 6 inhibitors, or combinations thereof. In some embodiments, the additional anticancer agent is an SHP2 inhibitor. Other additional anticancer agents are described herein.

[0243] Synthesis method The compounds described herein may be prepared from commercially available starting materials or synthesized using known organic, inorganic, or enzymatic processes.

[0244] The compounds of the present invention can be prepared by several methods well known to those skilled in the art of organic synthesis. For example, the compounds of the present invention can be synthesized by using the methods described in the following scheme in combination with synthetic methods known in the art of synthetic organic chemistry, or variations thereof as understood to those skilled in the art. These methods include, but are not limited to, the methods described in the following scheme.

[0245] The compounds in Tables 1 and 2 of this specification were prepared using the methods disclosed herein, or using the methods disclosed herein in combination with the knowledge of those skilled in the art. Scheme 1. General synthesis of functionalized bis-macrocyclic compounds

[0246] [ka]

[0247] A general overview of the synthesis of functionalized bis-macrocyclic compounds is shown in Scheme 1. A suitably substituted biaryl intermediate (1) can be prepared in one step from a suitably substituted 3-(5-bromo-2-iodo-1H-indole-3-yl)-2,2-dimethylpropan-1-ol intermediate and a suitably substituted methylpyrazine ester-containing arylboronic acid ester by palladium-mediated coupling followed by ester hydrolysis. Macrolactonization, followed by amine and phenol deprotection, can yield the macrocyclic ester (2).

[0248] A properly substituted 2-(tosyloxymethyl)-3-(amide)cyclic amine (3) can be prepared by coupling O-protected N-methyl-L-valine (4) with a properly substituted 2-(hydroxymethyl)-3-carboxylate cyclic amine using a peptide coupling reagent, followed by tosylation of the alcohol and deprotection of the carboxylic acid.

[0249] The final functionalized bis-macrocyclic compound can then be prepared by peptide coupling of the macrocyclic ester (1) with the intermediate (3), followed by macrocyclic ether formation in the presence of a base. Deprotection and coupling with an amine and a suitably substituted carboxylic acid (or other coupling partner) yield the bis-macrocyclic product (5).

[0250] Scheme 2. Alternative general synthesis of macrocyclic ester intermediate (2)

[0251] [ka]

[0252] Alternatively, macrocyclic ester intermediate (2) can be prepared as described in Scheme 2. A suitably substituted arylboronic acid ester (5) is coupled with a suitably protected 3-(5-bromo-indole-3-yl)-2,2-dimethylpropan-1-ol (6) in the presence of a palladium catalyst. This is followed by indole iodation, alcohol deprotection, and ester hydrolysis. Subsequent coupling with a methyl(S)-pyrazine ester, ester hydrolysis, and macrolactonization yield the iodized macrocyclic intermediate (7). Intermediate (2) is obtained by coupling with a suitably substituted arylboronic acid ester (8) in the presence of a palladium catalyst and N-alkylation of indole, followed by subsequent amine and phenol deprotection.

[0253] Scheme 3. General synthesis of functionalized amine bis-macrocyclic compounds

[0254] [ka]

[0255] A general overview of the synthesis of functionalized bis-macrocyclic compounds is shown in Scheme 3. A well-protected hydroxyalkyl amino acid can be coupled with O-protected N-methyl-L-valine (3) using a peptide coupling reagent. Subsequent alcohol and carboxylic acid deprotection can yield a well-substituted intermediate (7).

[0256] The protected amine bis-macrocyclic intermediate can be prepared by peptide coupling of the macrocyclic ester intermediate (2) with a carboxylic acid (7), followed by bis-macrocyclic ether formation in the presence of triphenylphosphine and azodicarboxylate. Deprotection and coupling of the amine with a appropriately substituted carboxylic acid (or other coupling partner) yield the final bis-macrocyclic product (8).

[0257] Scheme 4. General synthesis of functionalized amine bis-macrocyclic compounds

[0258] [ka]

[0259] A general overview of the synthesis of functionalized bis-macrocyclic compounds is shown in Scheme 4. A appropriately substituted terminal alkyne (9) can be coupled with an appropriately substituted iodized bromoarene (10) in the presence of a palladium catalyst. The macrocyclic intermediate (12) can then be obtained by reduction of the aryl alkyne intermediate (11), followed by amino acid N deprotection, carboxylic acid deprotection, macrocyclization in the presence of a peptide coupling reagent, ester hydrolysis, and peptide coupling with a methyl(S)-pyrazine ester. The functionalized bis-macrocyclic compound (13) can then be obtained by palladium-mediated coupling with an appropriately substituted 3-(5-boronate-indole-3-yl)-2,2-dimethylpropan-1-ol, macrolactonization, amine deprotection, and coupling of the amine with an appropriately substituted carboxylic acid (or other coupling partner).

[0260] Scheme 5. General Synthesis of Functionalized Amine-Bis-Macrocyclic Compounds

[0261] [ka]

[0262] A general overview of the synthesis of functionalized bis-macrocyclic compounds is shown in Scheme 5. A suitably substituted 2-bromo-4-bromomethyl-5-ethenyl five-membered heteroarene (14) can be reacted with ethyl 2-((diphenylethylene)amino) in the presence of a base and a chiral auxiliary. Subsequent amide coupling with a suitably substituted 2-(ethenyl)-3-(amide)cyclic amine (15), followed by olefin metathesis, ester hydrolysis, and amide coupling with a methyl(S)-pyrazine ester, can yield the macrocyclic compound (16).

[0263] The functionalized amine bis-macrocyclic compound (17) can then be obtained by palladium-mediated coupling with appropriately substituted 3-(5-boronate-indole-3-yl)-2,2-dimethylpropan-1-ol, methyl ester hydrolysis, macrolactonization, amine deprotection, and coupling of the amine with appropriately substituted carboxylic acid (or other coupling partners).

[0264] Scheme 6. General synthesis of functionalized amine bis-macrocyclic compounds

[0265] [ka]

[0266] A general overview of the synthesis of functionalized bis-macrocyclic compounds is shown in Scheme 6. Appropriately substituted iodized bromoalene (10) can be coupled with a vinylboronic acid ester in the presence of a palladium catalyst. Aldehyde (18) can be obtained by hydrolysis of the vinyl ether in the presence of an acid.

[0267] A suitable N-functionalized O-protected amino acid (19) can be coupled with an aldehyde (18) in the presence of an acid and a reducing agent. Subsequently, carboxylic acid deprotection and coupling of O-protected N-methyl-L-valine (3) in the presence of an amide coupling reagent can be carried out. Further deprotection of the carboxylate and amine, followed by cyclization and hydrolysis in the presence of a peptide coupling reagent, can yield a macrocyclic intermediate (20).

[0268] The appropriately substituted biaryl intermediate (21) can then be prepared in two steps by palladium-mediated coupling with the appropriately substituted intermediate (20) and 3-(5-boronate-indole-3-yl)-2,2-dimethylpropan-1-ol, followed by ester hydrolysis. Subsequent coupling with a methyl(S)-pyrazine ester using a peptide coupling reagent, ester hydrolysis, and macrolactonization can yield the functionalized amine bis-macrocyclic compound (22).

[0269] In any embodiment of this specification, the compounds of the present invention may be modified with substituents found in one or more of the following applications, by using the methods described in those applications in combination with methods provided herein and known to those skilled in the art: WO2024 / 060966, WO2024 / 017859, WO2024 / 008834, WO2024 / 008610, WO2023 / 232776, WO2023 / 208005, WO2023 / 086341, WO2023 / 025832, WO2023 / 015559, CN117720556, CN117720555, CN117720554, CN117534687, CN117534685, and CN117534684 (each of which is incorporated herein by reference in its entirety).

[0270] Pharmaceutical composition and method of use The compounds of interest of the present invention are Ras inhibitors and are useful in the treatment of cancer. Accordingly, one embodiment of the present invention provides a pharmaceutical composition comprising the compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient, as well as a method for preparing such a composition using the compound of the present invention.

[0271] As used herein, the term “pharmaceutical composition” refers to a compound such as the compound of the present invention, or a pharmacovigilant salt thereof, formulated with pharmacovigilant excipients.

[0272] In some embodiments, the compound is present in the pharmaceutical composition in a unit dose appropriate for administration in a therapeutic regime, for example, demonstrating a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, the pharmaceutical composition may be specifically formulated for administration in solid or liquid form, including, for example, oral administration, e.g., oral tablets (aqueous or nonaqueous solutions or suspensions), tablets, e.g., for buccal, sublingual, and systemic absorption, pills, powders, granules, and pastes for application to the tongue; parenteral administration, e.g., sterile solutions or suspensions, or controlled-release formulations, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection; topical application, e.g., creams, ointments, or controlled-release patches or sprays applied to the skin, lungs, or oral cavity; e.g., pessaries, creams, or foams, e.g., adapted for administration to the vagina or rectum; sublingual; ocular; transdermal; or nasal cavity, lungs, and other mucosal surfaces.

[0273] As used herein, “medically acceptable excipients” refers to any inert component (e.g., a vehicle capable of suspending or dissolving an active compound) that is non-toxic and non-inflammatory in the subject matter. Typical excipients include, for example, anti-tacks, antioxidants, binders, coatings, compression aids, disintegrants, pigments (colorants), softeners, emulsifiers, fillers (diluents), film-forming agents or coatings, flavorings, fragrances, flow enhancers (flow improvers), lubricants, preservatives, printing inks, adsorbents, suspending agents or dispersants, sweeteners, or hydration water. Excipients include, but are not limited to, optionally substituted butylated hydroxytoluene (BHT), calcium carbonate, dibasic calcium phosphate, calcium stearate, croscarmellose, cross-linked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, optionally substituted hydroxypropylcellulose, optionally substituted hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol. Those skilled in the art are familiar with a wide variety of drugs and materials useful as excipients.See, for example, Ansel, et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, et al., Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. In some embodiments, the composition comprises at least two different pharmaceutically acceptable excipients.

[0274] The compounds described herein may be provided or used in salt form, for example, pharmaceutically acceptable salt form, whether or not expressly defined otherwise, unless expressly defined to the contrary. As used herein, the term “pharmaceutically acceptable salt” refers to a salt of a compound described herein that is suitable for use in contact with human and other animal tissues without excessive toxicity, irritation, allergic reactions, etc., within the bounds of sound medical judgment, and that is commensurate with a reasonable benefit-risk ratio. pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and Pharmaceutical Salts: Properties, Selection, and Use, (Eds. PHStahl and CGWermuth), Wiley-VCH, 2008. Salts can be prepared in situ during the final isolation and purification of the compounds described herein, or separately by reacting a free base with a suitable organic acid.

[0275] The compounds of the present invention may have ionizable groups so that they can be prepared as pharmaceutically acceptable salts. These salts may be acid addition salts containing inorganic or organic acids, or, in the case of the acidic form of the compounds of the present invention, the salts may be prepared from inorganic or organic bases. In some embodiments, the compounds are prepared or used as pharmaceutically acceptable salts, which are prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases, such as hydrochloric acid, sulfuric acid, hydrobromic acid, acetic acid, lactic acid, citric acid, or tartaric acid for forming acid addition salts, and potassium hydroxide, sodium hydroxide, ammonium hydroxide, caffeine, various amines, etc. for forming basic salts, are well known in the art. Methods for preparing suitable salts are established in the art.

[0276] Typical acid addition salts include acetate, adipine, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphor sulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptone, glycerophosphate, hemisulfate, heptone, hexanoate, hydrobromide, hydrochloride, hydroiodide, and 2-(optionally substituted) hydroxyethanesulfate. These include rufonates, lactobionates, lactates, laurates, lauryl sulfates, malates, maleates, malons, methanesulfons, 2-naphthalenesulfons, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectins, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propions, stearates, succinates, sulfates, tartrates, thiocyans, toluenesulfons, undecanoic acid, valersates, etc. Typical alkali metal salts or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc., as well as non-toxic ammonium, quaternary ammonium, and amine cations (including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc.).

[0277] As used herein, the term “subject” refers to any member of the animal kingdom. In some embodiments, “subject” refers to a human at any stage of development. In some embodiments, “subject” refers to a human patient. In some embodiments, “subject” refers to a non-human animal. In some embodiments, a non-human animal is a mammal (e.g., rodents, mice, rats, rabbits, monkeys, dogs, cats, sheep, cattle, primates, or pigs). In some embodiments, subjects include, but are not limited to, mammals, birds, reptiles, amphibians, fish, or worms. In some embodiments, subjects may be transgenic animals, genetically modified animals, or clones.

[0278] As used herein, the term “dosage form” refers to a physically distinct unit of a compound (e.g., the compound of the present invention) for administration to a subject. Each unit contains a predetermined amount of the compound. In some embodiments, such amount is a unit dose (or a whole fraction thereof) appropriate for administration according to a dosing regimen (i.e., a therapeutic dosing regimen) that has been determined to correlate with a desired or beneficial outcome when administered to a relevant population. Those skilled in the art will understand that the total amount of a therapeutic composition or compound to be administered to a particular subject may be determined by one or more attending physicians and may involve administration in multiple dosage forms.

[0279] As used herein, the term “dosage regimen” refers to a set of unit doses (typically multiple) administered individually to a subject (typically at intervals of a certain period). In some embodiments, a given therapeutic compound (e.g., a compound of the present invention) has a recommended dosage regimen, which may consist of one or more doses. In some embodiments, the dosage regimen comprises multiple doses, each of which is spaced apart by a period of the same length, and in some embodiments, the dosage regimen comprises multiple doses, each individual dose spaced apart by at least two different periods of a certain length. In some embodiments, all doses in the dosage regimen are of the same unit dose. In some embodiments, different doses in the dosage regimen are of different amounts. In some embodiments, the dosage regimen comprises a first dose, followed by one or more additional doses, each being a second dose different from the first dose. In some embodiments, the dosage regimen comprises a first dose, followed by one or more additional doses, each being the same second dose as the first dose. In some embodiments, the dosing regimen correlates with a desired or beneficial outcome when administered across a relevant population (i.e., it is a therapeutic dosing regimen).

[0280] A "treatment regimen" refers to a regimen of administration that correlates with a desired or beneficial treatment outcome when administered across a relevant population. The term “treatment” (and in addition, “to treat” or “to treat”), in its broadest sense, refers to any administration of a substance (e.g., a compound of the present invention) that partially or completely reduces, improves, alleviates, inhibits, partially or completely delays the onset of, partially or completely reduces the severity of, or partially or completely reduces the incidence of one or more symptoms, features, or causes of a particular disease, disorder, or condition. In some embodiments, such treatment may be administered to a subject that does not exhibit signs of the disease, disorder, or condition, or to a subject that exhibits only early signs of the disease, disorder, or condition. Alternatively or additionally, in some embodiments, treatment may be administered to a subject that exhibits one or more established signs of the disease, disorder, or condition. In some embodiments, treatment may be administered to a subject that has been diagnosed with the disease, disorder, or condition. In some embodiments, the treatment may be for subjects known to have one or more susceptibility factors that are statistically correlated with an increased risk of progression of the disease, disorder, or condition.

[0281] The term “therapeutic dose” means an amount sufficient to treat a disease, disorder, or condition when administered to a population suffering from or susceptible to such disease, disorder, or condition, according to a therapeutic dosing regimen. In some embodiments, a therapeutic dose is one that reduces the incidence or severity of one or more symptoms of the disease, disorder, or condition, or delays the onset of one or more symptoms. Those skilled in the art will understand that the term “therapeutic dose” does not actually require that treatment success be achieved in a particular individual. Rather, a therapeutic dose may be an amount that, when administered to patients requiring such treatment, provides a particular desired pharmacological response in a significant number of subjects. It is specifically understood that certain subjects may actually be “refractory” to a “therapeutic dose.” In some embodiments, a reference to a therapeutic dose may refer to an amount measured in one or more specific tissues (e.g., tissues affected by the disease, disorder, or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine). Those skilled in the art will understand that in some embodiments, a therapeutic dose may be formulated or administered as a single dose. In some embodiments, the therapeutically effective dose may be formulated or administered in multiple doses, for example, as part of an administration regimen.

[0282] When used for the treatment of a target, the compounds of the present invention, or pharmaceutically acceptable salts thereof, may be formulated as pharmaceutical or veterinary compositions. Depending on the target being treated, the mode of administration, and the type of treatment desired, e.g., prevention, prophylaxis, or treatment, the compounds, or pharmaceutically acceptable salts thereof, may be formulated in a manner consistent with these parameters. A summary of such techniques can be found in Remington: The Science and Practice of Pharmacy, 21 stThis information can be found in Edition, Lippincott Williams & Wilkins, (2005), and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and JCBoylan, 1988–1999, Marcel Dekker, New York, each of which is incorporated herein by reference.

[0283] Each composition can be prepared according to conventional mixing, granulation, or coating methods, and the pharmaceutical composition may contain about 0.1% to about 99%, about 5% to about 90%, or about 1% to about 20% of the compound of the present invention or a pharmaceutically acceptable salt thereof, based on weight or volume. In some embodiments, the compounds described herein or a pharmaceutically acceptable salt thereof may be present in an amount totaling 1% to 95% based on the total weight of the composition, such as the pharmaceutical composition.

[0284] The composition may be provided in dosage forms suitable for intra-articular, oral, parenteral (e.g., intravenous, intramuscular), rectal, skin, subcutaneous, topical, transdermal, sublingual, nasal cavity, vagina, intravesical, intraurethral, ​​intrathecal, epidural, ear, or ocular administration, or by injection, inhalation, or direct contact with the mucous membranes of the nose, genitourinary tract, genitals, or oral cavity. Therefore, the pharmaceutical composition may be in the form of, for example, tablets, capsules, pills, powders, granules, suspensions, emulsions, liquids, gels containing hydrogels, pastes, ointments, creams, plasters, oral medications, osmotic delivery devices, suppositories, enemas, injections, implants, sprays, preparations suitable for iontophoresis delivery, or aerosols. The composition may be formulated according to conventional pharmaceutical regulations.

[0285] As used herein, the term “administration” means the administration of a composition (e.g., a compound described herein, or a preparation containing a compound described herein) to a subject or system. Administration to an animal subject (e.g., to a human) may be by any suitable route. For example, in some embodiments, administration may be by the bronchus (including by bronchial infusion), buccal, enteral, interdermal, arterial, intradermal, gastric, intramedullary, intramuscular, intranasal, intraperitoneal, intramedullary, intravenous, intraventricular, mucosa, nasal cavity, oral, rectal, subcutaneous, sublingual, topical, trachea (including by intratracheal infusion), percutaneous, vaginal, or vitreous humor.

[0286] Formulations may be prepared in a form suitable for systemic administration or topical or local administration. Systemic formulations may include those designed for injection (e.g., intramuscular, intravenous, or subcutaneous injection) or may be prepared for transdermal, transmucosal, or oral administration. Formulations generally include diluents, and optionally adjuvants, buffers, preservatives, etc. The compound, or a pharmaceutically acceptable salt thereof, may also be administered in liposome compositions or as particulate emulsions.

[0287] For injection, the formulation may be prepared in the conventional form of either a liquid solution or suspension, or as a solid form suitable for a solution or suspension in a liquid before injection, or as an emulsion. Suitable excipients include, for example, water, saline solution, dextrose, glycerol, etc. Such compositions may also contain amounts of non-toxic auxiliary substances such as wetting agents or emulsifiers, pH buffers, etc., such as sodium acetate, sorbitan monolaurate, etc.

[0288] Various sustained-release systems for drugs have also been devised. See, for example, U.S. Patent No. 5,624,677. Systemic administration may also include relatively non-invasive methods such as the use of suppositories, transdermal patches, transmucosal delivery, and intranasal administration. Oral administration is also suitable for the compounds of the present invention or pharmaceutically acceptable salts thereof. As will be understood in the art, preferred forms include syrups, capsules, and tablets.

[0289] Each compound described herein, or a pharmaceutically acceptable salt thereof, may be formulated in various ways known in the art. For example, the first and second agents of a combination therapy may be formulated together or separately. Other modalities of combination therapy are described herein.

[0290] Drugs formulated individually or separately may be packaged together as a kit. Non-limiting examples include, but are not limited to, kits containing two pills, pills and powders, suppositories and liquids in vials, two topical creams, etc. Kits may include optional components to assist in the administration of a unit dose to a subject, such as vials for reconstituting powder forms, syringes for injection, customized IV delivery systems, inhalers, etc. In addition, unit dose kits may include instructions for the preparation and administration of the composition. Kits may be manufactured as single-use unit doses for a particular subject, as multi-use doses for a specific subject (at a constant dose, or with varying potency of individual compounds or their pharmaceutically acceptable salts as treatment progresses), or kits may contain multi-use doses suitable for administration to multiple subjects ("bulk packaging"). The components of a kit may be assembled into cartons, blister packs, bottles, tubes, etc.

[0291] Preparations for oral use include tablets containing the active ingredient(s) in a mixture with non-toxic, pharmaceutically acceptable excipients. These excipients may include, for example, inert diluents or fillers (e.g., sucrose, sorbitol, sugars, mannitol, microcrystalline cellulose, starch including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate); granulators and disintegrants (e.g., cellulose derivatives including microcrystalline cellulose, starch including potato starch, croscarmellose sodium, alginate, or alginic acid); binders (e.g., sucrose, glucose, sorbitol, acacia, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, aluminum magnesium silicate, sodium carboxymethylcellulose, methylcellulose, optionally substituted hydroxypropylmethylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethylene glycol); as well as lubricants, flow enhancers, and anti-adhesion agents (e.g., magnesium stearate, zinc stearate, stearic acid, silica, hydrogenated vegetable oil, or talc). Other excipients that are acceptable as pharmaceuticals may include colorants, flavoring agents, plasticizers, humectants, buffering agents, etc.

[0292] Two or more compounds may be mixed or separated within a tablet, capsule, or other vehicle. In one example, the first compound may be contained inside the tablet and the second compound on the outside, thereby releasing a substantial portion of the second compound before the first compound.

[0293] Formulations for oral use may also be provided as chewable tablets, or as rigid gelatin capsules in which the active ingredient is mixed with an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate, or kaolin), or as soft gelatin capsules in which the active ingredient is mixed with a water or oil medium, such as peanut oil, liquid paraffin, or olive oil. Powders, granules, and pellets may be prepared using the components described above for tablets and capsules by conventional methods, for example, using a mixer, fluidized bed apparatus, or spray drying equipment.

[0294] Dissolution or diffusion-controlled release can be achieved by appropriate coating of a tablet, capsule, pellet, or granular formulation of the compound, or by incorporating the compound, or a pharmaceutically acceptable salt thereof, into a suitable matrix. A controlled-release coating may comprise one or more of the above-mentioned coating materials, or, for example, shellac, beeswax, glycowax, castor wax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryl distearate, glycerol palmitostearate, ethylcellulose, acrylic resin, dl-polylactic acid, cellulose acetate / butyrate, polyvinyl chloride, polyvinyl acetate, vinylpyrrolidone, polyethylene, polymethacrylate, methyl methacrylate, 2-(optionally substituted)hydroxyl methacrylate, methacrylate hydrogel, 1,3-butylene glycol, ethylene glycol methacrylate, or polyethylene glycol. In controlled-release matrix formulations, the matrix material may also include, for example, hydrated methylcellulose, carnauba wax and stearyl alcohol, Carbopol 934, silicone, glyceryl tristearate, methyl acrylate-methyl methacrylate, polyvinyl chloride, polyethylene, or halogenated fluorocarbons.

[0295] Liquid forms in which the compounds of the present invention, or pharmaceutically acceptable salts thereof, and compositions can be incorporated for oral administration include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and emulsions flavored with edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[0296] In general, when administered to humans, the oral dosage of any of the compounds of the present invention, or any pharmaceutically acceptable salt thereof, will depend on the properties of the compound, which can be easily determined by those skilled in the art. The dosage may be, for example, about 0.001 mg to about 2000 mg per day, about 1 mg to about 1000 mg per day, about 5 mg to about 500 mg per day, about 100 mg to about 1500 mg per day, about 500 mg to about 1500 mg per day, about 500 mg to about 2000 mg per day, or any range within these that can be derived.

[0297] In some embodiments, the pharmaceutical composition may further contain additional compounds having antiproliferative activity. Depending on the mode of administration, the compounds, or pharmaceutically acceptable salts thereof, will be formulated into a composition suitable for easy delivery. Each compound, or pharmaceutically acceptable salt thereof, of the combination therapy can be formulated in various ways known in the art. For example, the first and second agents of the combination therapy can be formulated together or separately. Preferably, the first and second agents are formulated together for simultaneous or near-simultaneous administration of the agents.

[0298] The compounds and pharmaceutical compositions of the present invention can be formulated and used in combination therapy, that is, the compounds and pharmaceutical compositions can be formulated together with one or more other desired therapeutic agents or medical procedures, or administered in parallel with them, before them, or after them. In a particular combination of therapies (therapeutic agents or procedures) used in a combination regimen, the suitability of the desired therapeutic agent or procedure and the desired therapeutic effect to be achieved will be taken into consideration. It will also be understood that the therapies used may achieve the desired effect against the same disorder, or they may achieve different effects (e.g., control of any adverse effects).

[0299] The administration of each drug in the combination therapies described herein may be independent, occurring 1 to 4 times daily for 1 day to 1 year, or even for the lifetime of the patient. Chronic long-term administration may be indicated.

[0300] How to use In some embodiments, the present invention discloses methods for treating a disease or disorder characterized by abnormal Ras activity resulting from a Ras variant. In some embodiments, the disease or disorder is cancer.

[0301] Accordingly, methods for treating cancer in subjects requiring treatment for cancer are also provided, the methods comprising administering to the subject a therapeutically effective amount of the compound of the present invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing such compound or salt. In some embodiments, the cancer is colorectal cancer, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, appendiceal cancer, melanoma, acute myeloid leukemia, small intestine cancer, papillary cancer, germ cell cancer, cervical cancer, cancer of unknown primary origin, endometrial cancer, gastroesophageal junction cancer, GI neuroendocrine cancer, ovarian cancer, sex cord-stromal tumor cancer, hepatobiliary cancer, or bladder cancer. In some embodiments, the cancer is appendiceal cancer, endometrial cancer, or melanoma. Methods for treating Ras protein-related disorders in subjects requiring treatment for Ras protein-related disorders are also provided, the methods comprising administering to the subject a therapeutically effective amount of the compound of the present invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing such compound or salt.

[0302] In some embodiments, the compounds of the present invention, or pharmaceutically acceptable salts thereof, pharmaceutical compositions comprising such compounds or salts, and methods provided herein may be used to treat a wide variety of cancers, including, for example, lung cancer, prostate cancer, breast cancer, brain tumors, skin cancer, cervical cancer, testicular cancer, and other tumors. More specifically, cancers that can be treated by the compounds of the present invention, or salts thereof, pharmaceutical compositions comprising such compounds or salts, and methods include, but are not limited to, astrocellular carcinoma, breast cancer, cervical cancer, colorectal cancer, endometrial cancer, esophageal cancer, gastric cancer, head and neck cancer, hepatocellular carcinoma, laryngeal cancer, lung cancer, oral cancer, ovarian cancer, prostate cancer, and thyroid cancer and sarcoma. Other cancers include, for example, the following:

[0303] The heart, for example, sarcomas (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyomas, fibromas, lipomas, and teratomas, Lung cancer, for example, bronchogenic carcinoma (squamous cell carcinoma, anaplastic small cell carcinoma, anaplastic large cell carcinoma, adenocarcinoma), alveolar (bronchial) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondrotoxic hamartoma, mesothelioma, The digestive tract, for example, the esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, islet cell adenoma, glucagonoma, gastrinoma, carcinoid tumor, vipoma), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, chorioadenoma, hamartoma, leiomyoma), The urogenital tract, for example, the kidneys (adenocarcinoma, Wilms' tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testes (seminocarcinoma, teratoma, fetal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenoid tumor, lipoma), The liver, for example, liver cancer (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma, Biliary tract, for example, gallbladder cancer, ampullary cancer, bile duct cancer, Bone, for example, osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulosarcoma), multiple myeloma, malignant giant cell tumor, chordoma, osteochondroma (osteochondrosis), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma, and giant cell tumor, Nervous system, for example, skull (osteoma, hemangioma, granuloma, xanthomas, degenerative osteitis), meninges (meningioma, meningiosarcoma, glioma), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germ cell tumor (pineal glandoma), glioblastoma multiforme, oligodendroglioma, Schwann cell tumor, retinoblastoma, congenital tumor), spinal cord cancer, neurofibromatosis type 1, meningioma, glioma, sarcoma), Gynecology, for example, the uterus (endometrial cancer, uterine cancer, endometrial cancer), cervix (cervical cancer, precancerous cervical dysplasia), ovaries (ovarian cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassifiable cancer), granulosa-theca cell tumor, Sertoli-Leydig cell tumor, undifferentiated germ cell tumor, malignant teratoma), vulva (squamous cell carcinoma, carcinoma in situ, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, staphyloid sarcoma (embryonic rhabdomyosarcoma), fallopian tube (carcinoma), Hematologic, for example, blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia), myeloproliferative disorders (for example, myelofibrosis and myeloproliferative neoplasms), multiple myeloma, myelodysplastic syndrome, Hodgkin's disease, non-Hodgkin lymphoma (malignant lymphoma), Skin, for example, malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, dysplastic nevi, lipoma, hemangioma, dermatofibroma, keloid, psoriasis, and Adrenal glands, for example, neuroblastoma.

[0304] In some embodiments, the Ras protein is wild-type (Ras WT Therefore, in some embodiments, the compounds of the present invention are Ras WT (For example, K-Ras WT H-Ras WT or N-Ras WT It is used in methods for treating patients with cancer including ). In some embodiments, the Ras protein is used in Ras amplification (e.g., K-Ras amp Therefore, in some embodiments, the compounds of the present invention are Ras amp (K-Ras amp H-Ras amp or N-Ras amp It is used in methods for treating patients with cancer including ). In some embodiments, the cancer includes Ras mutations such as the Ras mutations described herein. In some embodiments, the mutations are selected from the following:

[0305] (a) The following K-Ras mutants: G12D, G12V, G12C, G13D, G12R, G12A, Q61H, G12S, A146T, G13C, Q61L, Q61R, K117N, A146V, G12F, Q61K, L19F, Q22K, V14I, A59T, A146P, G13R, G12L, or G13V, and combinations thereof. (b) The following H-Ras variants: Q61R, G13R, Q61K, G12S, Q61L, G12D, G13V, G13D, G12C, K117N, A59T, G12V, G13C, Q61H, G13S, A18V, D119N, G13N, A146T, A66T, G12A, A146V, G12N, or G12R, and combinations thereof, as well as (c) The following N-Ras variants: Q61R, Q61K, G12D, Q61L, Q61H, G13R, G13D, G12S, G12C, G12V, G12A, G13V, G12R, P185S, G13C, A146T, G60E, Q61P, A59D, E132K, E49K, T50I, A146V, or A59T, and combinations thereof. or any combination of the above. In some embodiments, the cancer comprises a K-Ras mutation selected from the group consisting of G12C, G12D, G13C, G12V, G13D, G12R, G12S, Q61H, Q61K, Q61R, and Q61L. In some embodiments, the cancer comprises a K-Ras mutation that is Q61H. In some embodiments, the cancer comprises an N-Ras mutation selected from the group consisting of G12C, Q61H, Q61K, Q61L, Q61P, and Q61R. In some embodiments, the cancer comprises an H-Ras mutation selected from the group consisting of Q61H and Q61L. In some embodiments, the cancer comprises a K-Ras mutation that is Q61H. In some embodiments, the compounds of the present invention inhibit multiple Ras variants. In some embodiments, the compounds of the present invention inhibit one or more additional Ras variants in addition to Ras WT (For example, K-, H-, or N-Ras) WT And K-Ras G12D, G12V, G12C, G13D, G12R, G12A, Q61H, G12S, A146T, G13C, Q61L, Q61R, K117N, A146V, G12F, Q61K, L19F, Q22K, V14I, A59T, A146P, G13R, G12L, or G13V;K, H or N-Ras WTand H-Ras Q61R, G13R, Q61K, G12S, Q61L, G12D, G13V, G13D, G12C, K117N, A59T, G12V, G13C, Q61H, G13S, A18V, D119N, G13N, A146T, A66T, G12A, A146V, G12N, or G12R; or K, H or N-Ras WT and inhibits N-Ras Q61R, Q61K, G12D, Q61L, Q61H, G13R, G13D, G12S, G12C, G12V, G12A, G13V, G12R, P185S, G13C, A146T, G60E, Q61P, A59D, E132K, E49K, T50I, A146V, or A59T). In some embodiments, the compounds of the present invention inhibit Ras in addition to one or more additional Ras mutations. amp (For example, K-, H-, or N-Ras) amp And K-Ras G12D, G12V, G12C, G13D, G12R, G12A, Q61H, G12S, A146T, G13C, Q61L, Q61R, K117N, A146V, G12F, Q61K, L19F, Q22K, V14I, A59T, A146P, G13R, G12L, or G13V;K, H or N-Ras amp and H-Ras Q61R, G13R, Q61K, G12S, Q61L, G12D, G13V, G13D, G12C, K117N, A59T, G12V, G13C, Q61H, G13S, A18V, D119N, G13N, A146T, A66T, G12A, A146V, G12N, or G12R; or K, H or N-Ras amp It inhibits N-Ras Q61R, Q61K, G12D, Q61L, Q61H, G13R, G13D, G12S, G12C, G12V, G12A, G13V, G12R, P185S, G13C, A146T, G60E, Q61P, A59D, E132K, E49K, T50I, A146V, or A59T).

[0306] Methods for detecting Ras mutations are known in the art. Such methods include, but are not limited to, direct sequencing and the use of highly sensitive diagnostic assays (CE-IVD marked), such as those described in Domagala, et al., Pol J Pathol 3:145-164 (2012) (which is incorporated herein by reference), including TheraScreen PCR, AmoyDx, PNAClamp, RealQuality, EntroGen, LightMix, StripAssay, Hybcell plexA, Devyser, Surveyor, Cobas, and TheraScreen Pyro. See also, for example, WO2020 / 106640.

[0307] In some embodiments, the cancer is caused by the Ras Q61H mutation and TP53, STK11 LOF , CDKN2A, KEAP1, CDKN2B, MTAP, RBM10, SMARCA4, ATM, MYC, APC, SMAD4, PIK3CA, SOX9, FBXW7, PTEN, FLT3, AMER1, CDK8, AKT2, RNF43, GATA6, SF381, IGH, CDKN2C, DNMT3A, RB1, TRAF3, N-Ras, TET2, FAF1, BRAF, KMT2A, RUNX1, PTPN11, ETV6, NPM1, or MYH11 mutations. In some embodiments, the cancer is non-small cell lung cancer and includes the K-Ras Q61H mutation, and TP53, STK11 LOF, and include CDKN2A, KEAP1, CDKN2B, MTAP, RBM10, SMARCA4, ATM, or MYC mutations. In some embodiments, the cancer is colorectal cancer and includes the K-Ras Q61H mutation and APC, TP53, SMAD4, PIK3CA, SOX9, FBXW7, PTEN, FLT3, AMER1, or CDK8 mutations. In some embodiments, the cancer is pancreatic cancer and includes the K-Ras Q61H mutation and TP53, CDKN2A, CDKN2B, MTAP, SMAD4, ATM, AKT2, RNF43, GATA6, or SF381 mutations. In some embodiments, the cancer is multiple myeloma and includes the K-Ras Q61H mutation and IGH, TP53, CDKN2C, DNMT3A, RB1, TRAF3, N-Ras, TET2, FAF1, or BRAF mutations. In some embodiments, the cancer is acute myeloid leukemia and includes the K-Ras Q61H mutation and the N-Ras, KMT2A, FLT3, DNMT3A, RUNX1, PTPN11, TP53, ETV6, NPM1, or MYH11 mutation. In some embodiments, the cancer is melanoma and the Ras mutation includes the N-Ras mutation such as N-Ras Q61R or N-Ras Q61K. In any of the above, the compound is Ras WT (For example, K-, H-, or N-Ras) WT ) or Ras amp (For example, K-, H-, or N-Ras) amp ) can also be inhibited in a similar manner.

[0308] A method for inhibiting intracellular Ras protein is also provided, comprising contacting cells with an effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof. A method for inhibiting RAF-Ras binding is also provided, comprising contacting cells with an effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof. The cells may be cancer cells. The cancer cells may be of any type of cancer described herein. The cells may be in vivo or in vitro.

[0309] Combination therapy The method of the present invention may include the compound of the present invention, used alone or in combination with one or more additional therapies (e.g., non-pharmacological treatments or therapeutic agents). The dosage of one or more of the additional therapies (e.g., non-pharmacological treatments or therapeutic agents) may be reduced from the standard dosage when administered alone. For example, the dosage may be determined empirically from the combination and permutation of drugs, or it may be estimated by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6 (2005)).

[0310] The compounds of the present invention may be administered before, after, or in parallel with one or more of the additional therapies. When combined, the dosage of the compounds of the present invention and the dosage of one or more additional therapies (e.g., non-pharmacological treatments or therapeutic agents) provide a therapeutic effect (e.g., a synergistic or additive therapeutic effect). The compounds of the present invention and the additional therapies (e.g., anticancer agents) may be administered together (e.g., in a single pharmaceutical composition) or separately, and if administered separately, this may be done simultaneously or sequentially. Such sequential administrations may be close in time or far apart.

[0311] In some embodiments, additional therapy involves the administration of side effect inhibitors (e.g., drugs intended to reduce the occurrence or severity of side effects of treatment). For example, in some embodiments, the compounds of the present invention may also be used in combination with therapeutic agents for treating nausea. Examples of agents that may be used to treat nausea include dronabinol, granisetron, metoclopramide, ondansetron, and prochlorperazine, or pharmaceutically acceptable salts thereof.

[0312] In some embodiments, one or more additional therapies include non-pharmacological treatment (e.g., surgery or radiotherapy). In some embodiments, one or more additional therapies include therapeutic agents (e.g., compounds or biological agents that are anti-angiogenic agents, signaling inhibitors, antiproliferative agents, glycolysis inhibitors, or autophagy inhibitors). In some embodiments, one or more additional therapies include non-pharmacological treatment (e.g., surgery or radiotherapy) and therapeutic agents (e.g., compounds or biological agents that are anti-angiogenic agents, signaling inhibitors, antiproliferative agents, glycolysis inhibitors, or autophagy inhibitors). In other embodiments, one or more additional therapies include two therapeutic agents. In yet another embodiment, one or more additional therapies include three therapeutic agents. In some embodiments, one or more additional therapies include four or more therapeutic agents.

[0313] In this section on combination therapy, all references for the listed medications are incorporated by reference, regardless of whether they are explicitly stated otherwise. Non-pharmacological therapy Examples of non-pharmacological treatments include, but are not limited to, radiation therapy, cryotherapy, hyperthermia, surgery (e.g., surgical excision of tumor tissue), and T-cell adoptive transfer (ACT) therapy.

[0314] In some embodiments, the compounds of the present invention can be used as adjuvant therapy after surgery. In some embodiments, the compounds of the present invention can be used as neoadjuvant therapy before surgery.

[0315] Radiotherapy may be used in subjects (e.g., mammals (e.g., humans)) to inhibit abnormal cell proliferation or to treat hyperproliferative disorders such as cancer. Techniques for performing radiotherapy are known in the art. Radiotherapy may be performed by one or a combination of several methods, including, but not limited to, external beam radiation therapy, internal beam radiation therapy, implantable radiation therapy, stereotactic radiosurgery, total body radiation therapy, radiotherapy, and permanent or transient intratissue brachytherapy. As used herein, the term “brachytherapy” refers to radiotherapy delivered by spatially restricted radioactive material inserted into or near a tumor or other site of proliferative tissue disease. This term is intended to include, but not limited to, exposure to radioisotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, and radioisotopes of Lu). Suitable radiation sources for use as cell modifiers of the present invention include both solid and liquid forms. In non-limiting examples, the radiation source may be a radionuclide, e.g., I-125, I-131, Yb-169, Ir-192 as a solid source, I-125 as a solid source, or other radionuclides that emit photons, beta particles, gamma rays, or other therapeutic rays. The radioactive material may also be a fluid prepared from a solution of any radionuclide(s), e.g., a solution of I-125 or I-131, or the radioactive fluid may be produced using a suitable fluid slurry containing small particles of a solid radionuclide such as Au-198 or Y-90. Furthermore, the radionuclide(s) may be incorporated into a gel or radioactive microspheres.

[0316] In some embodiments, the compounds of the present invention can make abnormal cells more sensitive to radiotherapy for the purpose of killing or inhibiting the proliferation of such cells. Accordingly, the present invention further relates to a method for sensitizing abnormal cells in mammals to radiotherapy, the method comprising administering a certain amount of the compound of the present invention to a mammal, the amount being effective in sensitizing abnormal cells to radiotherapy. The amount of compound in this method can be determined according to the means for determining the effective amount of such compound described herein. In some embodiments, the compounds of the present invention may be used as adjuvant therapy after radiotherapy or as neoadjuvant therapy before radiotherapy.

[0317] In some embodiments, the non-pharmacological treatment is T cell adoptive transfer (ACT) therapy. In some embodiments, the T cells are activated T cells. The T cells can be modified to express a chimeric antigen receptor (CAR). CAR-modified T (CAR-T) cells can be generated by any method known in the art. For example, CAR-T cells can be generated by introducing a suitable expression vector encoding a CAR into T cells. The source of T cells is obtained from the subject before proliferation and genetic modification of the T cells. T cells can be obtained from several sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, umbilical cord blood, thymic tissue, tissue from the site of infection, ascites, pleural fluid, spleen tissue, and tumors. In certain embodiments of the present invention, any number of T cell lines available in the art can be used. In some embodiments, the T cells are autologous T cells. Whether before or after genetic modification of T cells to express a desired protein (e.g., CAR), T cells are, for example, U.S. Patent Nos. 6,352,694, 6,534,055, 6,905,680, 6,692,964, 5,858,358, 6,887,466, 6,905,681, and 7,144. It can be activated and propagated by generally using the methods described in Nos. 575, 7,067,318, 7,172,869, 7,232,566, 7,175,843, 7,572,631, 5,883,223, 6,905,874, 6,797,514, and 6,867,041.

[0318] Therapeutic drugs The therapeutic agent may be a compound used to treat cancer or related conditions. The compounds of the present invention may be combined with a second, third, or fourth therapeutic agent, or more therapeutic agents. The compounds of the present invention may be combined with one or more therapeutic agents in addition to one or more nonpharmacological therapies.

[0319] For example, the therapeutic agent may be a steroid. Steroids are known in the art. Therefore, in some embodiments, one or more additional therapies include steroids. Preferred steroids include 21-acetoxypregnenolone, alclomethasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, crocortol, cloprednol, corticosterone, cortisone, cortibazole, deflazacort, desonide, dexoxymethasone, dexamethasone, diflorasone, diflucortol, difprednate, enoxolone, fluazacort, fluchloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinonide, flucortin butyl, flucortolone, fluorometholone, fluperolone acetate, flupredniden acetate, fluprednisolone, flulandrenolide, This may include, but is not limited to, fluticasone propionate, formocortal, halcinonide, halobetazole propionate, halomethasone, hydrocortisone, loteprednol etavonate, mazipredone, medrisone, meprednisone, methylprednisolone, mometasone furoate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethylaminoacetate, sodium prednisolone phosphate, prednisone, prednival, prednylidene, rimexolone, thixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexaacetonide, and their salts or derivatives.

[0320] Further examples of therapeutic agents that may be used in combination therapy with the compounds of the present invention include the compounds described in the following patents: U.S. Patent Nos. 6,258,812, 6,630,500, 6,515,004, 6,713,485, 5,521,184, 5,770,599, 5,747,498, 5,990,141, 6,235,764, and 8,62 Patent No. 3,885, and international patent applications WO01 / 37820, WO01 / 32651, WO02 / 68406, WO02 / 66470, WO02 / 55501, WO04 / 05279, WO04 / 07481, WO04 / 07458, WO04 / 09784, WO02 / 59110, WO99 / 45009, WO00 / 59509, WO99 / 61422, WO00 / 12089, and WO00 / 02871.

[0321] The therapeutic agent may be a biological agent used to treat cancer or related conditions (e.g., cytokines (e.g., interferons, or interleukins such as IL-2)). Biological agents are known in the art. In some embodiments, the biological agent is an immunoglobulin-based biological agent, e.g., monoclonal antibodies (e.g., humanized antibodies, fully human antibodies, Fc fusion proteins, or functional fragments thereof) that stimulate an anti-cancer response by agonizing a target or antagonizing an antigen important to cancer. Antibody-drug conjugates are also included.

[0322] The therapeutic agent may be a T-cell checkpoint inhibitor. Such checkpoint inhibitors are known in the art. In one embodiment, the checkpoint inhibitor is an inhibitory antibody (e.g., a monospecific antibody such as a monoclonal antibody). The antibody may be, for example, a humanized or fully human antibody. In some embodiments, the checkpoint inhibitor is a fusion protein, for example, an Fc receptor fusion protein. In some embodiments, the checkpoint inhibitor is a drug such as an antibody that interacts with a checkpoint protein. In some embodiments, the checkpoint inhibitor is a drug such as an antibody that interacts with a ligand of a checkpoint protein. In some embodiments, the checkpoint inhibitor is a CTLA-4 inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) (e.g., an anti-CTLA-4 antibody or fusion protein). In some embodiments, the checkpoint inhibitor is a PD-1 inhibitor or antagonist (e.g., an inhibitory antibody or small molecule inhibitor). In some embodiments, the checkpoint inhibitor is a PD-L1 inhibitor or antagonist (e.g., an inhibitory antibody or small molecule inhibitor). In some embodiments, the checkpoint inhibitor is a PD-L2 inhibitor or antagonist (e.g., an inhibitory antibody or Fc fusion or small molecule inhibitor) (e.g., a PD-L2 / Ig fusion protein). In some embodiments, the checkpoint inhibitor is an inhibitor or antagonist (e.g., an inhibitory antibody or small molecule inhibitor) of B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, B-7 family ligands, or combinations thereof.In some embodiments, the checkpoint inhibitor is pembrolizumab, nivolumab, PDR001 (NVS), REGN2810 (Sanofi / Regeneron), PD-L1 antibody, e.g., avelumab, durvalumab, atezolizumab, pizilizumab, JNJ-63723283 (JNJ), BGB-A317 (BeiGene & Celgene), or Preusser, M. et al. These are checkpoint inhibitors disclosed in al. (2015) Nat. Rev. Neurol. (including, but not limited to, ipilimumab, tremelimumab, nivolumab, pembrolizumab, AMP224, AMP514 / MEDI0680, BMS936559, MEDl4736, MPDL3280A, MSB0010718C, BMS986016, IMP321, lirirumab, IPH2101, 1-7F9, and KW-6002).

[0323] The therapeutic agent may be an anti-TIGIT antibody, such as MBSA43, BMS-986207, MK-7684, COM902, AB154, MTIG7192A, or OMP-313M32 (ethigirimab). Other anti-TIGIT antibodies are known in the art.

[0324] Therapeutic agents may be drugs that treat cancer or related conditions (e.g., cytotoxic agents, non-peptide small molecules, or other compounds useful for treating cancer or related conditions, collectively referred to as “anticancer agents”). Anticancer agents may be, for example, chemotherapeutic agents or targeted therapy agents. Such agents are known in the art.

[0325] Anticancer agents include mitotic inhibitors, intercalation antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, bioresponse modifiers, alkylating agents, antimetabolites, folic acid analogs, pyrimidine analogs, purine analogs and related inhibitors, vinca alkaloids, epipodophyllotoxin, antibiotics, L-asparaginase, topoisomerase inhibitors, interferons, platinum coordination complexes, anthracendione-substituted ureas, methylhydrazine derivatives, corticosteroids, progestins, estrogens, anti-estrogens, androgens, anti-androgens, and gonadotropin-releasing hormone analogs. Further anticancer agents include leucovorin (LV), irenotecan, oxaliplatin, capecitabine, paclitaxel, and doxetaxel. In some embodiments, one or more additional therapies comprise two or more anticancer agents. Two or more anticancer drugs can be used in a cocktail for combined administration or administered separately. Preferred administration regimens for combination anticancer drugs are known in the art and are described, for example, in Saltz et al., Proc.Am.Soc.Clin.Oncol.18:233a(1999) and Douillard et al., Lancet 355(9209):1041-1047(2000).

[0326] Other non-exclusive examples of anticancer drugs include Gleevec® (imatinib mesylate); Kyprolis® (carfilzomib); Velcade® (bortezomib); Casodex® (bicalutamide); Iressa® (gefitinib); alkylating agents, e.g., thiotepa and cyclophosphamide; alkyl sulfonates, e.g., busulfan, improsulfan, and pigosulfan; aziridines, e.g., benzodopa, carbocone, metsuredopa, and uredopa; ethyleneimines and methylamelamines (including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolomelamine); acetogenins (especially bratacin and bratacinone); camptothecin (including the synthetic analog topotecan); briostatin; calistatin; CC-1065 (its adzeresin, carzeresin, and including bizeresin synthetic analogs); cryptophycin (especially cryptophycin 1 and cryptophycin 8); drastatin; duocalmycin (including synthetic analogs KW-2189 and CB1-TM1); eryuterobin; pancratistatin; sarcodictiin A; spongistatin; nitrogen mustard, e.g., chlorambucil, chlornafadin, colophosphamide, estramustine, ifosfamide, mechloretamine, mechloretamine Electrolyte dihydrochloride, melphalan, nobembitine, fenestrine, prednimustine, trophosphamide, and uracil mustard; nitrosourea, e.g., camulstine, chlorozotosine, fotemustine, lomustine, nimustine, and ranimustine; antibiotics, e.g., engine antibiotics (e.g., calicheamicin, e.g., calicheamicin gamma II and calicheamicin omega II (see, e.g., Agnew, Chem. Intl. Ed Engl. 33:183-186 (1994)); dynemicin, e.g., dynemicin A; bisphosphonates, e.g., clodronate; esperamicin;Neocardinostatin chromophore and related pigment proteins enediin antibiotic chromophore, acrasinomycin, actinomycin, autoramycin, azaserin, bleomycin, kactinomycin, calichemycin, carabicin, kaminomycin, carminomycin, cardinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, adriamycin (doxorubicin), morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, de Oxidoxorubicin, epirubicin, esorubicin, idarubicin, marcelomycin, mitomycin, e.g., mitomycin C, mycophenolic acid, nogaramycin, olibomycin, peplomycin, potophyllomycin, puromycin, queramycin, rhodorubicin, streptonigrin, streptozocin, tubercidine, ubenimex, dinostatin, zolbicin; antimetabolites, e.g., methotrexate and 5-fluorouracil (5-FU); folic acid analogs, e.g., denopterin, pteropterin, trimethotrexate; Phosphorus analogs, e.g., fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs, e.g., ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens, e.g., carsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; anti-adrenal agents, e.g., aminoglutethimide, mitotane, trilostane; folic acid supplements, e.g., floric acid; acetaminophen Glutton; Aldophosphamide glycoside; Aminolevulinic acid; Enyluracil; Amsacrin; Bestrabusil; Bisanthren; Edatraxate; Defofamine; Demecolsin; Diadicone; Elfomitin; Erliptinium acetate; Epotilon, e.g., Epotilon B; Etoglucid; Gallium nitrate; Hydroxyurea; Lentinan; Ronidamine; Maytansinoids, e.g., Maytansin and Ansamitosin; Mitoguazone; Mitoxanthrone; Mopidamol; Nitracrine; Pentostatin; Fenamet; Pirarubicin; Rosoxanthrone;Podophyllic acid; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, OR); razoxane; rhizoxin; schizophyllan; spirogermanium; tenuazonic acid; triadiquan; 2,2',2''-trichlorotriethylamine; trichothecenes, e.g., T-2 toxin, beracrine A, loridine A and anguidin; urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitractol; pipobromane; gasitosine; arabinoside ("Ara-C"); cyclophosphatidyl Mido; thiotepa; taxoids, e.g., Taxol® (paclitaxel), Abraxane® (a cremophore-free albumin-modified nanoparticle formulation of paclitaxel), and Taxotere® (doxetaxel); chlorambucil; tamoxifen (Nolvadex®); raloxifene; aromatase inhibitor 4(5)-imidazole; 4-hydroxytamoxifen; trioxyfen; keoxyfen; LY11 7018; Onapristone; Toremifene (Fareston®); Flutamide, Niltamide, Bicalutamide, Leuprolide, Goserelin; Chlorambucil; Gemzar® Gemcitabine; 6-Thiogunine; Mercaptopurine; Platinum-coordinated complexes, e.g., cisplatin, oxaliplatin and carboplatin; Vinblastine; Platinum; Etoposide (VP-16); Ifosfamide; Mitoxanthrone; Vincristine; Navelbi Examples include ne(registered trademark) (vinorelbine); novantrone; teniposide; edatrexate; daunomycin; aminopterin; ibandronate; irinotecan (e.g., CPT-11); topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoids, e.g., retinoic acid; esperamicin; capecitabine (e.g., Xeloda(registered trademark)); and any pharmaceutically acceptable salts of the above.

[0327] Non-exclusive examples of additional anticancer drugs include trastuzumab (Herceptin®), bevacizumab (Avastin®), cetuximab (Erbitux®), rituximab (Rituxan®), Taxol®, Arimidex®, ABVD, Avisin, avagovomab, acridine carboxamide, adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, alfarazine, arbocidib, 3-aminopyridine-2-carboxyaldehyde thiosemicarbazone, amonafide, anthracendione, anti-CD22 immunotoxin, antitumor drugs (e.g., cell cycle nonspecific antitumor drugs, and other antitumor drugs described herein), antitumor herbs, apadicon, atiprimod, azathioprine, berotecan, and Damustine, BIBW2992, Bilicodal, Brostalysin, Briostatin, Butionine sulfoximine, CBV (chemotherapy), Kallikrin, Dichloroacetate, Discodermorid, Elsamitrusine, Enocitabine, Eribulin, Exatecan, Exislind, Ferginol, Forodesine, Phosfestrol, ICE chemotherapy regimen, IT-101, Imexon, Imiquimod, Examples include indolocarbazole, ilofluben, lanikidal, larotaxel, lenalidomide, lucanton, lulutotecan, maphosphamide, mitozolomid, napoxidine, nedaplatin, olaparib, ortataxel, PAC-1, pawpaw, pixantrone, proteasome inhibitors, rebeccamycin, reximod, rubitecan, SN-38, salinosporamide A, sapacitabine, Stanford V, swainsonin, talaporfin, talikidal, tegafur-uracil, temodal, tesetaxel, triplatin tetranitrate, tris(2-chloroethyl)amine, troxacitabine, uramustine, bajimezan, vinflunin, ZD6126, and zoskidal.

[0328] Further non-limiting examples of anticancer drugs include natural products, e.g., vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine), epipodophyllotoxins (e.g., etoposide and teniposide), antibiotics (e.g., dactinomycin (actinomycin D), daunorubicin, and idarubicin), anthracyclines, mitoxantrone, bleomycin, plicamycin (mitramycin), mitomycin, and enzymes (e.g., those that systemically metabolize L-asparagine). In addition, L-asparaginase, which leads to a deficiency in cells that lack the ability to synthesize their own asparagine, antiplatelet agents, antiproliferative / antimitotic alkylating agents, such as nitrogen mustard (e.g., mechloretamine, cyclophosphamide and analogs, melphalan, and chlorambucil), ethyleneimine and methylmelamine (e.g., hexamethylmelamine and thiotepa), CDK inhibitors (e.g., CDK4 / 6 inhibitors, such as abemaciclib, ribociclib, and palbociclib;Cericiclib (UCN-01, P1446A-05, PD-0332991, Dinacyclib (P27-00, AT-7519, RGB286638, and SCH727965)), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) and its analogs, as well as streptozocin), trazeneth-dacarbazine (DTIC), antiproliferative / antimitotic antimetabolites (e.g., folate analogs, pyrimidine analogs (e.g., fluorouracil, floxuridine, and cytarabine)), purine analogs and related inhibitors (e.g., mercaptopurine, thioguanine, pentostatin, and 2-chlorodeoxyadenosine)), aromatase inhibitors (e.g., anastrozole, exemestane, and letrozole), and platinum-coordinate complexes (e.g., cisplatin and Carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide, histone deacetylase (HDAC) inhibitors (e.g., trichostatin, sodium butyrate, apicidan, suberoylanilide hydroxamic acid, vorinostat, bellinostat, LBH589, romidepsin, ACY-1215, and panobinostat), mTOR inhibitors (e.g., bisutucertib, tem Sirolimus, everolimus, ridaflorimus, and sirolimus), KSP(Eg5) inhibitors (e.g., Array520), DNA binders (e.g., Zalypsis®), PI3K inhibitors, e.g., PI3K delta inhibitors (e.g., GS-1101 and TGR-1202), PI3K delta and gamma inhibitors (e.g., CAL-130), copanlisib, alpelisib, and idelalisib;Multi-kinase inhibitors (e.g., TG02 and sorafenib), hormones (e.g., estrogen) and hormone agonists, e.g., luteinizing hormone-releasing hormone (LHRH) agonists (e.g., goserelin, leuprolide and triptorelin), BAFF neutralizing antibodies (e.g., LY2127399), IKK inhibitors, p38MAPK inhibitors, anti-IL-6 (e.g., CNT0328), telomerase inhibitors (e.g., GRN163L), aurora kinase inhibitors (e.g., MLN8237), cell surface monoclonal antibodies (e.g., anti-CD38 (HUMAX-CD38), anti-CSl (e.g., elotuzumab), HSP90 inhibitors ( Examples include 17AAG and KOS953), P13K / Akt inhibitors (e.g., perifosine), Akt inhibitors (e.g., GSK-2141795), PKC inhibitors (e.g., Enzastaurin), FTIs (e.g., Zanestra®), anti-CD138 (e.g., BT062), Torc1 / 2-specific kinase inhibitors (e.g., INK128), ER / UPR targeting agents (e.g., MKC-3946), cFMS inhibitors (e.g., ARRY-382), JAK1 / 2 inhibitors (e.g., CYT387), PARP inhibitors (e.g., olaparib and veliparib (ABT-888)), and BCL-2 antagonists.

[0329] In some embodiments, the anticancer agent is selected from mechloretamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, Navelbine®, sorafenib, or any analogue or derivative variant described above.

[0330] In some embodiments, the anticancer agent is a HER2 inhibitor. HER2 inhibitors are known in the art. Non-limiting examples of HER2 inhibitors include monoclonal antibodies, e.g., trastuzumab (Herceptin®) and pertuzumab (Perjeta®); small molecule tyrosine kinase inhibitors, e.g., gefitinib (Iressa®), erlotinib (Tarceva®), pyritinib, CP-654577, CP-724714, canertinib (CI 1033), HKI-272, lapatinib (GW-572016; Tykerb®), PKI-166, AEE788, BMS-599626, HKI-357, BIBW2992, ARRY-334543, and JNJ-26483327.

[0331] In some embodiments, the anticancer agent is an ALK inhibitor. ALK inhibitors are known in the art. Non-limiting examples of ALK inhibitors include ceritinib, TAE-684 (NVP-TAE694), PF02341066 (crizotinib or 1066), alectinib, brigatinib, entrectinib, ensartinib (X-396), lorlatinib, ASP3026, CEP-37440, 4SC-203, TL-398, PLB1003, TSR-011, CT-707, TPX-0005, and AP26113. Additional examples of ALK kinase inhibitors are described in Examples 3-39 of WO05016894.

[0332] In some embodiments, the anticancer agent is an inhibitor of a downstream member of receptor tyrosine kinase (RTK) / growth factor receptor (e.g., SHP2 inhibitors (e.g., SHP099, TNO155, RMC-4550, RMC-4630, JAB-3068, JAB-3312, RLY-1971, ERAS-601SHP3809, PF-07284892, or BBP-398) or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, etc. These include tautomers, SOS1 inhibitors (e.g., BI-1701963, BI-3406, SDR5, BAY-293, MRTX-0902, or RMC-5845, or pharmaceutically acceptable salts, solvates, isomers (e.g., stereoisomers), prodrugs, or tautomers thereof), Raf inhibitors, MEK inhibitors, ERK inhibitors, PI3K inhibitors, PTEN inhibitors, AKT inhibitors, or mTOR inhibitors (e.g., mTORC1 inhibitors or mTORC2 inhibitors). In some embodiments, the anticancer agent is JAB-3312.

[0333] In some embodiments, the anticancer agent is an SOS1 inhibitor. SOS1 inhibitors are known in the art. In some embodiments, the SOS1 inhibitor is WO2022219035, WO2022214594, WO2022199670, WO2022146698, WO2022081912, WO2022058344, WO2022026465, WO2022017519, WO2021173524, WO2021130731, WO2021127429, WO2021092115, WO2021105960 Selected from those disclosed in WO2021074227, WO2020180768, WO2020180770, WO2020173935, WO2020146470, WO2019201848, WO2019122129, WO2018172250, and WO2018115380, or pharmaceutically acceptable salts, solvates, isomers (e.g., stereoisomers), prodrugs, or tautomers thereof.

[0334] In some embodiments, the anticancer agent is an additional Ras inhibitor or Ras vaccine, or another therapeutic modality designed to directly or indirectly reduce the oncogenic activity of Ras. Such agents are known in the art. In some embodiments, the anticancer agent is an additional Ras inhibitor. In some embodiments, the Ras inhibitor targets Ras in an active state, i.e., a GTP-bound state. In some embodiments, the Ras inhibitor targets Ras in an inactive state, i.e., a GDP-bound state. In some embodiments, the Ras inhibitor is, for example, an inhibitor of K-Ras G12C, e.g., AMG. These include 510, MRTX1257, MRTX849, JNJ-74699157, LY3499446, ARS-1620, ARS-853, BPI-421286, LY3537982, JDQ443, JAB-3312, JAB-21822, JAB-21000, IBI351, ERAS-3490, RMC-6291, BI1823911, D-1553, D3S-001, HBI-2438, HS-10370, MK-1084, YL-15293, BBO-8520 (ON / OFF inhibitor), FMC-376 (ON / OFF inhibitor), GEC255, or GDC-6036. In some embodiments, the Ras inhibitor is a K-Ras G12D inhibitor, such as MRTX1133, JAB-22000, MRTX282, ERAS-4, HRS-4642, BI-2852, ASP3082, TH-Z827, TH-7835, RMC-9805, GFH375 (VS-7375), INCB161734, and KD-8. In some embodiments, the Ras inhibitor is a K-Ras G12V inhibitor, such as JAB-23000. In some embodiments, the KRAS(OFF) inhibitor is a pan-KRAS(OFF) inhibitor. In specific embodiments, the pan-KRAS(OFF) inhibitor is JAB-23400, JAB-23425, BI-2493, BI-2865, QTX-3034 (G12D preference), QTX3544 (G12V preference), ZG2001, BBO-a, BBO-B, or pan-KRas-IN-1. In some embodiments, the Ras inhibitor is JAB-23400. In some embodiments, the Ras inhibitor isRMC-6236 is one such example. In some embodiments, the Ras inhibitor is selected from the Ras(ON) inhibitors (i.e., Ras in a GTP-bound state) disclosed below (all of which are incorporated herein by reference), or pharmaceutically acceptable salts, solvates, isomers (e.g., stereoisomers), prodrugs, or tautomers thereof: WO2022 / 235870, WO2022 / 235864, WO2022 / 060836, WO2021091982, WO2021091967, WO2021091956, and WO2020132597. Other examples of Ras inhibitors are known in the art and include, for example, the following (all of which are incorporated herein by reference): WO2023287896, WO2023287730, WO2023284881, WO2023284730, WO2023284537, WO2023283933, WO2023283213, WO2023280960, WO2023280280, WO2 023278600, WO2023280136, WO2023280026, WO2023278600, WO2023274383, WO2023274324, WO2023034290, WO20 23020523, WO2023020521, WO2023020519, WO2023020518, WO2023018812, WO2023018810, WO2023018809, WO2023 018699, WO2023015559, WO2023014979, WO2023014006, WO2023010121, WO2023009716, WO2023009572, WO20230 04102, WO2023003417, WO2023001141, WO2023001123, WO2022271923, WO2022271823, WO2022271810, WO2022271 658, WO2022269508, WO2022266167, WO2022266069, WO2022266015, WO2022265974, WO2022261154, WO202226115 4, WO2022251576, WO2022251296, WO2022237815, WO2022232332, WO2022232331, WO2022232320, WO2022232318,WO2022223037、WO2022221739、WO2022221528、WO2022221386、WO2022216762、WO2022192794、WO2022192790、WO2022188729、WO2022187411、WO2022184178、WO2022173870、WO2022173678、WO2022135346、WO2022133731、WO2022133038、WO2022133345、WO2022132200、WO2022119748、WO2022109485、WO2022109487、WO2022066805、WO2022002102、WO2022002018、WO2021259331、WO2021257828、WO2021252339、WO2021248095、WO2021248090、WO2021248083、WO2021248082、WO2021248079、WO2021248055、WO2021245051、WO2021244603、WO2021239058、WO2021231526、WO2021228161、WO2021219090、WO2021219090、WO2021219072、WO2021218939、WO2021217019、WO2021216770、WO2021215545、WO2021215544、WO2021211864、WO2021190467、WO2021185233、WO2021180181、WO2021175199、2021173923、WO2021169990、WO2021169963、WO2021168193、WO2021158071、WO2021155716、WO2021152149、WO2021150613、WO2021147967、WO2021147965、WO2021143693、WO2021142252、WO2021141628、WO2021139748、WO2021139678、WO2021129824、WO2021129820、WO2021127404、WO2021126816、WO2021126799、WO2021124222、WO2021121371、WO2021121367、WO2021121330、WO2020050890、WO2020047192、WO2020035031、WO2020028706, WO2019241157, WO2019232419, WO2019217691, WO2019217307, WO2019215203, W O2019213526, WO2019213516, WO2019155399, WO2019150305, WO2019110751, WO2019099524, WO2 019051291, WO2018218070, WO2018217651, WO2018218071, WO2018218069, WO2018206539, WO201 8143315, WO2018140600, WO2018140599, WO2018140598, WO2018140514, WO2018140513, WO20181 40512, WO2018119183, WO2018112420, WO2018068017, WO2018064510, WO2017201161, WO2017172 979, WO2017100546, WO2017087528, WO2017058807, WO2017058805, WO2017058728, WO201705890 2, WO2017058792, WO2017058768, WO2017058915, WO2017015562, WO2016168540, WO2016164675, WO2016049568, WO2016049524, WO2015054572, WO2014152588, WO2014143659, and WO2013155223.

[0335] In some embodiments, therapeutic agents that may be combined with the compounds of the present invention are inhibitors of the MAP kinase (MAPK) pathway (or "MAPK inhibitors"). Such agents are known in the art. MAPK inhibitors include, but are not limited to, one or more MAPK inhibitors described in Cancers (Basel) 2015 Sep, 7(3):1758-1784. For example, MAPK inhibitors include trametinib, binimetinib, selumetinib, cobimetinib, LErafAON (NeoPharm), ISIS 5132; vemurafenib, pimacertib, TAK733, RO4987655 (CH4987655); CI-1040; PD-0325901; CH5126766; MAP855; AZD6244; refametinib (RDEA119 / BAY86-9766); GDC-0973 / XL581; AZD8330 (ARRY-424704 / ARRY-704); RO5126766 (Roche (PLoS One. 2014 Nov One or more of the following may be selected: (as described in 25;9(11)) and GSK1120212 (or JTP-74057 (as described in Clin Cancer Res. 2011 Mar 1;17(5):989-1000)). The MAPK inhibitor may be PLX8394, LXH254, GDC-5573, or LY3009120.

[0336] In some embodiments, the anticancer agent is an interferant or inhibitor of the RAS-RAF-ERK, PI3K-AKT-TOR, or PI3K-AKT signaling pathway. Such agents are known in the art. PI3K / AKT inhibitors may include, but are not limited to, one or more PI3K / AKT inhibitors described in Cancers (Basel) 2015 Sep;7(3):1758-1784. For example, the PI3K / AKT inhibitor may be selected from one or more of NVP-BEZ235, BGT226, XL765 / SAR245409, SF1126, GDC-0980, PI-103, PF-04691502, PKI-587, and GSK2126458.

[0337] In some embodiments, the anticancer agent is a PD-1 or PD-L1 antagonist. Such agents are known in the art. In some embodiments, additional therapeutic agents include ALK inhibitors, HER2 inhibitors, EGFR inhibitors, IGF-1R inhibitors, MEK inhibitors, PI3K inhibitors, AKT inhibitors, TOR inhibitors, MCL-1 inhibitors, BCL-2 inhibitors, SHP2 inhibitors, proteasome inhibitors, and immunotherapies. In some embodiments, additional therapeutic agents include FGFR inhibitors, PARP inhibitors, BET inhibitors, PRMT5i inhibitors, MAT2A inhibitors, VEGF inhibitors, and HDAC inhibitors. In some embodiments, the therapeutic agent may be a pan-RTK inhibitor, such as afatinib.

[0338] IGF-1R inhibitors are known in the art and include lincitinib or its pharmaceutically acceptable salts. EGFR inhibitors are known in the art and include, but are not limited to, small molecule antagonists, antibody inhibitors, or certain antisense nucleotides or siRNAs. Useful antibody inhibitors of EGFR include cetuximab (Erbitux®), panitumumab (Vectibix®), zaltumumab, nimotuzumab, and matuzumab. Further antibody-based EGFR inhibitors include any anti-EGFR antibody or antibody fragment that can partially or completely block EGFR activation by its native ligand. Non-exclusive examples of antibody-based EGFR inhibitors include those described in Modjtahedi et al., Br.J. Cancer 1993, 67:247-253, Teramoto et al., Cancer 1996, 77:639-645, Goldstein et al., Clin. Cancer Res. 1995, 1:1311-1318, Huang et al., 1999, Cancer Res. 15:59(8):1935-40, and Yang et al., Cancer Res. 1999, 59:1236-1243. The EGFR inhibitor may be a monoclonal antibody Mab E7.6.3 (Yang, 1999 (see above)), or Mab C225 (ATCC accession number HB-8508), or an antibody or antibody fragment having binding specificity thereto.

[0339] Small molecule antagonists of EGFR include gefitinib (Iressa®), erlotinib (Tarceva®), and lapatinib (TykerB®). See, for example, Yan et al., Pharmacogenetics and Pharmacogenomics in Oncology Therapeutic Antibody Development, BioTechniques 2005, 39(4):565-8, and Paez et al., EGFR Mutations in Lung Cancer Correlation with Clinical Response to Gefitinib Therapy, Science 2004, 304(5676):1497-500. In some embodiments, the EGFR inhibitor is osimertinib (Tagrisso®). Further non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in the following patent publications, and all pharmaceutically acceptable salts of such EGFR inhibitors: EP0520722, EP0566226, WO96 / 33980, U.S. Patent No. 5,747,498, WO96 / 30347, EP0787772, WO97 / 30034, WO97 / 30044, WO97 / 38994, WO97 / 49688, EP837063, WO98 / 02434, WO97 / 38983, WO95 / 19774, WO95 / 19970, WO97 / 13771, WO98 / 02437, WO98 / 02 438, WO97 / 32881, DE19629652, WO98 / 33798, WO97 / 32880, WO97 / 32880, EP682027, WO 97 / 02266, WO97 / 27199, WO98 / 07726, WO97 / 34895, WO96 / 31510, WO98 / 14449, WO98 / 14 450, WO98 / 14451, WO95 / 09847, WO97 / 19065, WO98 / 17662, U.S. Patent No. 5,789,427, U.S. Patent No. 5,650,415, U.S. Patent No. 5,656,643, WO99 / 35146, WO99 / 35132, WO99 / 07701, and WO92 / 20642.Additional non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in Traxler et al., Exp. Opin. Ther. Patents 1998, 8(12):1599-1625. In some embodiments, EGFR inhibitors are ERBB inhibitors. In humans, the ERBB family includes HER1 (EGFR, ERBB1), HER2 (NEU, ERBB2), HER3 (ERBB3), and HER (ERBB4).

[0340] MEK inhibitors are known in the art and include, but are not limited to, pimacertib, selumetinib, cobimetinib (Cotellic®), trametinib (Mekinist®), and binimetinib (Mektovi®). In some embodiments, the MEK inhibitor targets a MEK mutation which is a class I MEK1 mutation selected from D67N, P124L, P124S, and L177V. In some embodiments, the MEK mutation is a class II MEK1 mutation selected from ΔE51-Q58, ΔF53-Q58, E203K, L177M, C121S, F53L, K57E, Q56P, and K57N.

[0341] PI3K inhibitors are known in the art and include woltmannin; 17-hydroxywoltmannin analog described in WO06 / 044453; 4-[2-(1H-indazole-4-yl)-6-[[4-(methylsulfonyl)piperazine-1-yl]methyl]thieno[3,2-d]pyrimidine-4-yl]morpholine (also known as pictilisib or GDC-0941, described in WO09 / 036082 and WO09 / 055730); 2-methyl-2-[4-[3-methyl-2-oxo-8-(quinoline-3-yl)-2,3- Dihydroimidazo[4,5-c]quinoline-1-yl]phenyl]propionitrile (also known as BEZ235 or NVP-BEZ235, as described in WO06 / 122806); (S)-1-(4-((2-(2-aminopyrimidine-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidine-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (as described in WO08 / 070740); LY294002(2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (Axon Available from Medchem); PI103 hydrochloride (3-[4-(4-morpholinylpyrido-[3',2':4,5]flo[3,2-d]pyrimidine-2-yl]phenol hydrochloride (Available from Axon Medchem); PIK75 (2-methyl-5-nitro-2-[(6-bromoimidazo[1,2-a]pyridine-3-yl)methylene]-1-methylhydrazide-benzenesulfonic acid, monohydrochloride) (Available from Axon Medchem); PIK90 (N-(7,8-dimethoxy-2,3-dihydroimidazo[1,2-c]quinazolin-5-yl)-nicotinamide (Axon Available from Medchem); AS-252424 (5-[1-[5-(4-fluoro-2-hydroxy-phenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidined-2,4-dione (available from Axon Medchem); TGX-221 (7-methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido-[1,2-a]pyrimidine-4-one (available from Axon Medchem); XL-765;Other PI3K inhibitors include, but are not limited to, XL-147 and CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, BKM120, XL147, XL765, Palomid 529, GSK1059615, ZSTK474, PWT33597, IC87114, TGI00-115, CAL263, PI-103, GNE-477, CUDC-907, and AEZS-136.

[0342] AKT inhibitors are known in the art and include Akt-1-1 (inhibits Akt1) (Barnett et al., Biochem.J.2005,385(Pt.2):399-408); Akt-1-1,2 (inhibits Ak1 and 2) (Barnett et al., Biochem.J.2005,385(Pt.2):399-408); API-59CJ-Ome (e.g., Jin et al., Br.J.Cancer 2004,91:1808-12); 1-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO05 / 011700); indole-3-carbinol and its derivatives (e.g., U.S. Patent No. 6,656,963, Sarkar and Li J Nutr.2004,134(12)). Suppl):3493S-3498S); Perifosine (e.g., interfering with the membrane localization of Akt; Dasmahapatra et al. Clin. Cancer Res. 2004, 10(15):5242-52); Phosphatidylinositol ether lipid analogs (e.g., Gills and Dennis Expert. Opin. Investig. Drugs 2004, 13:787-97); and trisirivine (TCN or API-2 or NCI identifier: NSC 154020; Yang et al., Cancer Res. 2004, 64:4394-9), but not limited to these.

[0343] mTOR inhibitors are known in the art and include ATP-competitive mTORC1 / mTORC2 inhibitors, e.g., PI-103, PP242, PP30; Torin 1; FKBP12 enhancers; 4H-1-benzopyran-4-one derivatives; and rapamycin (also known as sirolimus) and its derivatives (Temsirolimus (Torisel®); Everolimus (Afinitor®, WO94 / 09010); Ridaforolimus (also known as deforolimus or AP23573); Rapalogs, e.g., as disclosed in WO98 / 02441 and WO01 / 14387). For example, AP23464 and AP23841; 40-(2-hydroxyethyl)rapamycin; 40-[3-hydroxy(hydroxymethyl)methylpropanoate]-rapamycin (also known as CC1779); 40-epi-(tetrazolyl)-rapamycin (also known as ABT578); 32-deoxorapamycin; 16-pentinyloxy-32(S)-dihydrorapamycin; derivatives disclosed in WO05 / 005434; National Patent Nos. 5,258,389, 5,118,677, 5,118,678, 5,100,883, 5,151,413, 5,120,842, and 5,256,790, as well as WO94 / 090101, WO92 / 05179, WO93 / 111130, WO94 / 02136, WO94 / 02485, WO95 / 14023, WO94 / 02136, WO95 / 16691, WO96 Derivatives disclosed in / 41807, WO96 / 41807, and WO2018204416; as well as phosphorus-containing rapamycin derivatives (e.g., WO05 / 016252), are included but not limited thereto. In some embodiments, the mTOR inhibitor is a bisteric inhibitor (see e.g., WO2018204416, WO2019212990, and WO2019212991), e.g., structure

[0344] [ka]

[0345] This is RMC-5552, which has [the following characteristics]. BRAF inhibitors that can be used in combination with the compounds of the present invention are known in the art and include, for example, vemurafenib, dabrafenib, and encorafenib. BRAF may include class 3 BRAF mutations. In some embodiments, class 3 BRAF mutations in human BRAF are selected from one or more of the following amino acid substitutions: D287H, P367R, V459L, G466V, G466E, G466A, S467L, G469E, N581S, N581I, D594N, D594G, D594A, D594H, F595L, G596D, G596R, and A762E.

[0346] MCL-1 inhibitors are known in the art and include, but are not limited to, AMG-176, MIK665, and S63845. The myeloid cell leukemia-1 (MCL-1) protein is one of the major anti-apoptotic members of the B-cell lymphoma-2 (BCL-2) protein family. Overexpression of MCL-1 is closely associated with tumor progression, as well as resistance to conventional chemotherapy, and to targeted therapies, including BCL-2 inhibitors such as ABT-263.

[0347] In some embodiments, additional therapeutic agents are SHP2 inhibitors. SHP2 inhibitors are known in the art. SHP2 is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene that contributes to several cellular functions, including proliferation, differentiation, maintenance of the cell cycle, and migration. SHP2 has two N-terminal Src homology domains (N-SH2 and C-SH2), a catalytic domain (PTP), and a C-terminal tail. These two SH2 domains regulate the intracellular localization and functional regulation of SHP2. The molecule exists in an inactive, autoinhibitory conformation stabilized by a binding network involving residues from both the N-SH2 and PTP domains. For example, stimulation by cytokines or growth factors acting via receptor tyrosine kinases (RTKs) exposes the catalytic site, leading to enzymatic activation of SHP2.

[0348] SHP2 is involved in signaling via the RAS-mitogenic factor-activated protein kinase (MAPK), JAK-STAT, or phosphoinositol 3-kinase-AKT pathways. Mutations in the PTPN11 gene, and subsequently in SHP2, have been identified in several human developmental disorders, including Noonan syndrome and Leopard syndrome, as well as in juvenile myelomonocytic leukemia, neuroblastoma, melanoma, acute myeloid leukemia, and human cancers such as breast cancer, lung cancer, and colon cancer. Some of these mutations destabilize the autoinhibitory conformation of SHP2, promoting SHP2 autoactivation or enhanced growth factor-driven activation. Therefore, SHP2 represents a very attractive target for the development of novel therapies for the treatment of various diseases, including cancer. SHP2 inhibitors (e.g., RMC-4550 or SHP099) combined with RAS pathway inhibitors (e.g., MEK inhibitors) have been shown to inhibit the growth of several cancer cell lines (e.g., pancreatic cancer, lung cancer, ovarian cancer, and breast cancer) in vitro. Therefore, combination therapy including SHP2 inhibitors and RAS pathway inhibitors can be a common strategy for preventing tumor resistance in a wide range of malignancies.

[0349] Non-limiting examples of such SHP2 inhibitors known in the art include Chen et al. Mol Pharmacol. 2006, 70, 562, Sarver et al., J.Med.Chem. 2017, 62, 1793, Xie et al., J.Med.Chem. 2017, 60, 113734, and Igbe et al. al., Oncotarget, 2017, 8, 113734, and PCT applications WO2023282702, WO2023280283, WO2023280237, WO2023018155, WO2023011513, WO2022271966, WO2022271964, WO2022271911, WO2022259157, WO2022242767, WO2022241975, WO2022237676, WO2022237367, WO20 22237178, WO2022235822, WO20222084008, WO2022135568, WO2021176072, WO2021171261, WO2021149817, WO2021148010, WO20 21147879, WO2021143823, WO2021143701, WO2021143680, WO2021121397, WO2021119525, WO2021115286, WO2021110796, WO2021 088945, WO2021073439, WO2021061706, WO2021061515, WO2021043077, WO2021033153, WO2021028362, WO2021033153, WO20210 28362, WO2021018287, WO2020259679, WO2020249079, WO2020210384, WO2020201991, WO2020181283, WO2020177653, WO2020165 734, WO2020165733, WO2020165732, WO2020156243, WO2020156242, WO2020108590, WO2020104635, WO2020094104, WO202009401 8, WO2020081848, WO2020073949, WO2020073945, WO2020072656, WO2020065453, WO2020065452, WO2020063760, WO2020061103,WO2020061101、WO2020033828、WO2020033286、WO2020022323、WO2019233810、WO2019213318、WO2019183367、WO2019183364、WO2019182960、WO2019167000、WO2019165073、WO2019158019、WO2019152454、WO2019051469、WO2019051084、WO2018218133、WO2018172984、WO2018160731、WO2018136265、WO2018136264、WO2018130928、WO2018129402、WO2018081091、WO2018057884、WO2018013597、WO2017216706、WO2017211303、WO2017210134、WO2017156397、WO2017100279、WO2017079723、WO2017078499、WO2016203406、WO2016203405、WO2016203404、WO2016196591、WO2016191328、WO2015107495、WO2015107494、WO2015107493、WO2014176488、WO2014113584、CN115677661、CN115677660、CN115611869、CN115521305、CN115490697、CN115466273、CN115394612、CN115304613、CN115304612、CN115300513、CN115197225、CN114957162、CN114920759、CN114716448、CN114671879、CN114539223、CN114524772、CN114213417、CN114195799、CN114163457、CN113896710、CN113248521、CN113248449、CN113135924、CN113024508、CN112920131、CN112823796、CN112409334、CN112402385、CN112174935、111848599、CN111704611、CN111393459、CN111265529、CN110143949、CN108113848、US11179397、US11044675、Examples include US11034705, US11033547, US11001561, US10988466, US10954243, US10934302, or US10858359, or their pharmaceutically acceptable salts, solvates, isomers (e.g., stereoisomers), prodrugs, or tautomers (each of these documents is incorporated herein by reference in its entirety).

[0350] In some embodiments, the SHP2 inhibitor binds to the active site. In some embodiments, the SHP2 inhibitor is a mixed-type irreversible inhibitor. In some embodiments, the SHP2 inhibitor binds to the allosteric site (e.g., a non-covalent allosteric inhibitor). In some embodiments, the SHP2 inhibitor is a covalent SHP2 inhibitor, such as an inhibitor that targets a cysteine ​​residue (C333) outside the active site of the phosphatase. In some embodiments, the SHP2 inhibitor is a reversible inhibitor. In some embodiments, the SHP2 inhibitor is an irreversible inhibitor. In some embodiments, the SHP2 inhibitor is SHP099. In some embodiments, the SHP2 inhibitor has the following structure:

[0351] [ka]

[0352] The SHP2 inhibitor is TNO155, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is RMC-4550. In some embodiments, the SHP2 inhibitor has the following structure:

[0353] [ka]

[0354] RMC-4630 having the following structure, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor has the following structure

[0355] [ka]

[0356] JAB-3068 having, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is JAB-3312. In some embodiments, the SHP2 inhibitor is the following compound,

[0357] [ka]

[0358] or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor has the following structure

[0359] [ka]

[0360] RLY-1971, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is ERAS-601. In some embodiments, the SHP2 inhibitor is BBP-398.

[0361] In some embodiments, additional therapeutic agents are selected from the group consisting of MEK inhibitors, HER2 inhibitors, SHP2 inhibitors, CDK4 / 6 inhibitors, mTOR inhibitors, SOS1 inhibitors, and PD-L1 inhibitors. See, for example, Hallin et al., Cancer Discovery, DOI:10.1158 / 2159-8290 (October 28, 2019) and Canon et al., Nature, 575:217 (2019). In some embodiments, the Ras inhibitor of the present invention is used in combination with a MEK inhibitor and an SOS1 inhibitor. In some embodiments, the Ras inhibitor of the present invention is used in combination with a PD-L1 inhibitor and an SOS1 inhibitor. In some embodiments, the Ras inhibitor of the present invention is used in combination with a PD-L1 inhibitor and an SHP2 inhibitor. In some embodiments, the Ras inhibitor of the present invention is used in combination with a MEK inhibitor and an SHP2 inhibitor. In some embodiments, the Ras inhibitor of the present invention is used in combination with an SHP2 inhibitor and a Ras inhibitor that inhibits multiple Ras isoforms and / or variants (e.g., RMC-6236). In some embodiments, the cancer is lung cancer, and the treatment comprises administering the Ras inhibitor of the present invention in combination with a second or third therapeutic agent (e.g., an SHP2 inhibitor and a Ras inhibitor that inhibits multiple Ras isoforms and / or variants). In some embodiments, the cancer is colorectal cancer, and the treatment comprises administering the Ras inhibitor of the present invention in combination with a second or third therapeutic agent (e.g., an SHP2 inhibitor and a Ras inhibitor that inhibits multiple Ras isoforms and / or variants). In some embodiments, the Ras inhibitor of the present invention is used in combination with immunotherapy and optionally with chemotherapeutic agents.

[0362] Proteasome inhibitors are known in the art and include, but are not limited to, carfilzomib (Kyprolis®), bortezomib (Velcade®), and oprozomib.

[0363] Immunotherapy includes, but is not limited to, monoclonal antibodies, immunomodulatory imides (IMiDs), GITR agonists, genetically modified T cells (e.g., CAR-T cells), bispecific antibodies (e.g., BiTE), and anti-PD-1, anti-PD-L1, anti-CTLA4, anti-LAGl, and anti-OX40 agents. Other immunotherapies are known in the art.

[0364] Immunomodulators (IMiDs) are a class of immunomodulatory drugs (drugs that modulate the immune response) that contain an imide group. The IMiD class includes thalidomide and its analogues (lenalidomide, pomalidomide, and apremilast).

[0365] Exemplary anti-PD-1 antibodies and their uses are described in Goldberg et al., Blood 2007, 110(1):186-192, Thompson et al., Clin. Cancer Res. Cancer Res. 2007, 13(6):1757-1761, and WO06 / 121168 A1), and also elsewhere in this specification.

[0366] FGFR inhibitors are known in the art, such as pemigatinib and erdafitinib (including FGFR2 and FGFR4 inhibitors). See, for example, Cancers (Basel), 2021 Jun;13(12)2968.

[0367] BET inhibitors are known in the art and include, for example, romidepsin, panobinostat, and bellinostat. See, for example, British J. Cancer 124:1478 (2021).

[0368] PRMT5i inhibitors are known in the art, for example, PF-0693999, PJ-68, and MRTX1719. See, for example, Biomed.Pharmacotherapy 144:112252 (2021).

[0369] MAT2A inhibitors are known in the art, for example, AG-270 and IDE397. See, for example, Exp Opin Ther Patents (2022) DOI:10.1080 / 13543776.2022.2119127.

[0370] GITR agonists include GITR fusion proteins and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies), e.g., GITR fusion proteins described in U.S. Patent No. 6,111,090, No. 8,586,023, WO2010 / 003118, and WO2011 / 090754; or e.g., U.S. Patent No. 7,025,962, EP1947183, U.S. Patent No. 7,812,135, U.S. Patent No. 8,388,967, U.S. Patent No. 8,59 This includes, but is not limited to, the anti-GITR antibodies described in U.S. Patent No. 1,886, U.S. Patent No. 7,618,632, EP1866339, and WO2011 / 028683, WO2013 / 039954, WO05 / 007190, WO07 / 133822, WO05 / 055808, WO99 / 40196, WO01 / 03720, WO99 / 20758, WO06 / 083289, WO05 / 115451, and WO2011 / 051726.

[0371] Another example of a therapeutic agent that may be used in combination with the compounds of the present invention is an anti-angiogenic agent. Anti-angiogenic agents are known in the art and include, but are not limited to, chemical compositions, antibodies, antigen-binding domains, radionuclides, and combinations and conjugates thereof, which are synthetically prepared in vitro. Anti-angiogenic agents may be agonists, antagonists, allosteric modulators, toxins, or more generally, act to inhibit or stimulate their targets (e.g., by activating or inhibiting receptors or enzymes), thereby potentially promoting cell death or halting cell proliferation. In some embodiments, one or more additional therapies include an anti-angiogenic agent.

[0372] Anti-angiogenic agents may include MMP-2 (matrix-metalloproteinase 2) inhibitors, MMP-9 (matrix-metalloproteinase 9) inhibitors, and COX-II (cyclooxygenase 11) inhibitors. Non-exclusive examples of anti-angiogenic agents include rapamycin, temsirolimus (CCI-779), everolimus (RAD001), sorafenib, sunitinib, and bevacizumab. Examples of useful COX-II inhibitors include arecoxib, valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors include WO96 / 33172, WO96 / 27583, WO98 / 07697, WO98 / 03516, WO98 / 34918, WO98 / 34915, WO98 / 33768, WO98 / 30566, WO90 / 05719, WO99 / 52910, and WO99 / 5 As described in 2889, WO99 / 29667, WO99007675, EP0606046, EP0780386, EP1786785, EP1181017, EP0818442, EP1004578, and US20090012085, and U.S. Patents 5,863,949 and 5,861,510. Preferred MMP-2 and MMP-9 inhibitors are those with little or no activity to inhibit MMP-1. More preferably are those that selectively inhibit MMP-2 or AMP-9 compared to other matrix metalloproteinases (i.e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13). Some specific examples of MMP inhibitors are AG-3340, RO32-3555, and RS13-0830.

[0373] Further exemplary anti-angiogenic agents include KDR (kinase domain receptor) inhibitors (e.g., antibodies and antigen-binding regions that specifically bind to kinase domain receptors), anti-VEGF agents (e.g., antibodies or antigen-binding regions that specifically bind to VEGF, or their soluble VEGF receptors or ligand-binding regions (e.g., bevacizumab)), e.g., VEGF-TRAP®, and anti-VEGF receptor agents (e.g., antibodies or antigen-binding regions that specifically bind thereto), VEGF inhibitors, EGFR inhibitors (e.g., antibodies or antigen-binding regions that specifically bind thereto), e.g., Vectibix® (panitumumab), erlotinib (Tarceva®), anti-Ang1 and anti-Ang2 agents (e.g., antibodies or antigen-binding regions that specifically bind to them or their receptors (e.g., Tie2 / Tek)), and anti-Tie2 kinase inhibitors (e.g., antibodies or antigen-binding regions that specifically bind thereto). Other anti-angiogenic agents include Campath, IL-8, B-FGF, Tek antagonists (US2003 / 0162712, US6,413,932), anti-Tweak agents (e.g., antibodies or antigen-binding domains that specifically bind, or soluble Tweak receptor antagonists; see US6,727,225), ADAM disintegrin domains that antagonize the binding of integrins to their ligands (US2002 / 0042368), and anti-eph receptors or anti-ephrin antibodies or antigens that specifically bind. These include binding regions (U.S. Patents 5,981,245, 5,728,813, 5,969,110, 6,596,852, 6,232,447, 6,057,124, and their respective patent family members), anti-PDGF-BB antagonists (e.g., antibody or antigen-binding regions that specifically bind), antibody or antigen-binding regions that specifically bind to PDGF-BB ligands, and PDGFR kinase inhibitors (e.g., antibody or antigen-binding regions that specifically bind to them). Additional anti-angiogenic agents include SD-7784 (Pfizer, USA); sirengitide (Merck KGaA, Germany, EPO0770622); pegaptanib octasodium (Gilead Sciences, USA);Alpha-statin (BioActa, UK); M-PGA (Celgene, USA, US5712291); Ilostat (Arriva, USA, US5892112); Emaxanib (Pfizer, USA, US5792783); Batalanib (Novartis, Switzerland); 2-Methoxyestradiol (EntreMed, USA); TLC ELL-12 (Elan, Ireland); Anecoltab acetate (Alcon, USA); Alpha-D148Mab (Amgen, USA); CEP-7055 (Cephalon, USA); Anti-Vn Mab (Crucell, Netherlands), DAC anti-angiogenic agent (ConjuChem, Canada); Angiocidin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko, Japan); SU-0879 (Pfizer, USA); CGP-79787 (Novartis, Switzerland, EP0970070); ARGENT technology (Ariad, USA); YIGSR-Stealth (Johnson & Johnson, USA); Fibrinogen-E fragment (BioActa, UK); Angiogenesis inhibitor (Trigen, UK); TBC-1635 (Encysive Pharmaceuticals, USA); SC-236 (Pfizer, USA); ABT-567 (Abbott, USA); Metastatin (EntreMed, USA); Maspin (Sosei, Japan); 2-Methoxyestradiol (Oncology Sciences Corporation, USA); ER-68203-00 (IV AX, USA); BeneFin (Lane Labs, USA); Tz-93 (Tsumura, Japan); TAN-1120 (Takeda, Japan); FR-111142 (Fujisawa, Japan, JP02233610); Platelet factor 4 (RepliGen, USA, EP407122); Vascular endothelial growth factor antagonist (Borean, Denmark); Bevacizumab (pINN) (Genentech, USA); Angiogenesis inhibitor (SUGEN, USA); XL784 (Exelixis, USA); XL647 (Exelixis, USA);MAb, alpha-5 beta-3 integrin, second generation (Applied Molecular Evolution, USA and Medlmmune, USA); Enzastaurin hydrochloride (Lilly, USA); CEP7055 (Cephalon, USA and Sanofi-Synthelabo, France); BC1 (Genoa Institute of Cancer Research, Italy); rBPI21 and BPI-derived anti-angiogenic agents (XOMA, USA); PI88 (Progen, Australia); Silengitide (Merck KGaA, Germany; Munich Technical University, Germany, Scripps Clinic and Research Foundation, USA); AVE8062 (Ajinomoto, Japan); AS1404 (Cancer Research Laboratory, New Zealand); SG292 (Telios, USA); Endostatin (Boston Childrens Hospital, USA); ATN161 (Attenuon, USA); 2-Methoxyestradiol (Boston Childrens Hospital) Hospital, USA); ZD6474 (AstraZeneca, UK); ZD6126 (Angiogene Pharmaceuticals, UK); PPI2458 (Praecis, USA); AZD9935 (AstraZeneca, UK); AZD2171 (AstraZeneca, UK); Batalanib (pINN) (Novartis, Switzerland and Schering AG, Germany); Tissue factor pathway inhibitor (EntreMed, USA); Pegaptanib (Pinn) (Gilead Sciences, USA); Xantolulizole (Yonsei University, South Korea); Vaccine, gene-based, VEGF-2 (Scripps Clinic and Research Foundation, USA); SPV5.2 (Supratek, Canada); SDX103 (University of California at San Diego, USA); PX478 (ProlX, USA);METASTATIN (EntreMed, USA); Troponin I (Harvard University, USA); SU6668 (SUGEN, USA); OXI4503 (OXiGENE, USA); o-Guanidine (Dimensional Pharmaceuticals, USA); Motupolamine C (British Columbia University, Canada); CDP791 (Celltech Group, UK); Atiprimod (pINN) (GlaxoSmithKline, UK); E7820 (Eisai, Japan); CYC381 (Harvard University, USA); AE941 (Aeterna, Canada); Vaccine, Angiogenesis (EntreMed, USA); Urokinase Plasminogen Activator Inhibitor (Dendreon, USA); Ogluphanide (pINN) (Melmotte, USA); HIF-Ralfa Inhibitor (Xenova, UK); CEP5214 (Cephalon, USA); BAY RES2622 (Bayer, Germany); Angiocidin (InKine, USA); A6 (Angstrom, USA); KR31372 (Korea Research Institute of Chemical Technology, South Korea); GW2286 (GlaxoSmithKline, UK); EHT0101 (ExonHit, France); CP868596 (Pfizer, USA); CP564959 (OSI, USA); CP547632 (Pfizer, USA); 786034 (GlaxoSmithKline, UK); KRN633 (Kirin Brewery, Japan); Drug delivery systems, intraocular, 2-methoxyestradiol; Anguinex (Maastricht University, Netherlands, and Minnesota) University, USA); ABT510 (Abbott, USA); AAL993 (Novartis, Switzerland); VEGI (ProteomTech, USA); Tumor necrosis factor-alpha inhibitors; SU11248 (Pfizer, USA and SUGEN USA); ABT518 (Abbott, USA);YH16 (Yantai Rongchang, China); S-3APG (Boston Children's Hospital, USA and EntreMed, USA); MAb, KDR (ImClone Systems, USA); MAb, Alpha 5 Beta (Protein Design, USA); KDR kinase inhibitor (Celltech Group, UK and Johnson & Johnson, USA); GFB116 (South Florida University, USA and Yale University, USA); CS706 (Sankyo, Japan); Comblestatin A4 prodrug (Arizona State University, USA); Chondroitinase AC (IBEX, Canada); BAY RES2690 (Bayer, Germany); AGM1470 (Harvard University, USA, Takeda, Japan and TAP, USA); AG13925 (Agouron, USA); Tetrathiomolybdate (University of Michigan, USA); GCS100 (Wayne State University, USA) CV247 (Ivy Medical, UK); CKD732 (Chong Kun Dang, South Korea); Ilsogladine (Nippon Shinyaku, Japan); RG13577 (Aventis, France); WX360 (Wilex, Germany); Squalamine (Genaera, USA); RPI4610 (Sirna, USA); Heparanase inhibitor (InSight, Israel); KL3106 (Kolon, South Korea); Honokiol (Emory University, USA); ZK CDK (Schering AG, Germany); ZK Angio (Schering AG, Germany); ZK229561 (Novartis, Switzerland, and Schering AG, Germany); XMP300 (XOMA, USA); VGA1102 (Taisho, Japan); VE-Cadherin-2 antagonist (ImClone Systems, USA);Vasostatin (National Institutes of Health, USA); Flk-1 (ImClone Systems, USA); TZ93 (Tsumura, Japan); TumStatin (Beth Israel Hospital, USA); Sclerated soluble FLT1 (vascular endothelial growth factor receptor 1) (Merck & Co, USA); Tie-2 ligand (Regeneron, USA); and thrombospondin 1 inhibitor (Allegheny Hea; This includes the Education and Research Foundation (USA).

[0374] Further examples of therapeutic agents that may be used in combination with the compounds of the present invention include agents (e.g., antibodies, antigen-binding domains, or soluble receptors) that specifically bind to and inhibit the activity of growth factors, such as hepatocyte growth factor (HGF, also known as scatter factor) antagonists, as well as antibodies or antigen-binding domains that specifically bind to the receptor c-Met. Such agents are known in the art.

[0375] Another example of a therapeutic agent that may be used in combination with the compounds of the present invention is an autophagy inhibitor. Autophagy inhibitors are known in the art and include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil®), bafilomycin A1, 5-amino-4-imidazole carboxamidriboside (AICAR), okadaic acid, autophagy-suppressing algal toxins that inhibit type 2A or type 1 protein phosphatases, analogs of cAMP, and drugs that increase cAMP levels, such as adenosine, LY204002, N6-mercaptopurine riboside, and vinblastine. Furthermore, antisense or siRNAs that inhibit the expression of proteins including, but not limited to, ATG5 (associated with autophagy) may also be used. In some embodiments, one or more additional therapies include an autophagy inhibitor.

[0376] Another example of therapeutic agents that can be used in combination with the compounds of the present invention is antitumor agents, which are known in the art. In some embodiments, one or more additional therapies include antitumor agents. Non-limiting examples of antitumor agents include acemannan, acralubicin, aldesleukin, alemtuzumab, alitretinoin, altretamine, amiphostine, aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole, ancer, ancestim, algravin, arsenic trioxide, BAM-002 (Novelos), bexarotene, bicalutamide, bromodeoxyuridine, capecitabine, cermoloukin, cetrorelix, cladribine, clotrimazole, cytarabine ocphosphate, and DA 3030 (Dong-A), daclizumab, denileukin difutox, deslorerin, dexrazoxane, dilazep, docetaxel, docosanol, doxelcalciferol, doxifluridine, doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HIT diclofenac, interferon alpha, daunorubicin, doxorubicin, tretinoin, ederfosine, edrecolomab eflornithine, emiteflu, epirubicin, epoetin beta, etoposide phosphate, exemestane, exislind, fadrozol, filgrastim, finasteride, fludarabine phosphate, formestan, fotemustine, gallium nitrate, gemcitabine, gemtuzumab zogamicin, gimeracil / oteracil / Gafur combination, glycopine, goserelin, heptaplatin, human chorionic gonadotropin, human fetal alpha-fetoprotein, ibandronic acid, idarubicin (imiquimod), interferon alpha, interferon alpha, natural type, interferon alpha-2, interferon alpha-2a, interferon alpha-2b, interferon alpha-N1, interferon alpha-n3, interferon alpha-con-1, interferon alpha, natural type, interferon beta, interferon beta-1a, interferon beta-1b, interferon gamma, natural type interferon gamma-1a, interferon gamma-1b, interleukin-1 beta, iobenguan,Irinotecan, Ilsogladine, Lanreotide, LC9018 (Yakult), Leflunomide, Lenograstim, Lentinan sulfate, Letrozole, Leukocyte alpha interferon, Leuprorelin, Levamizole + Fluorouracil, Rialozol, Lovaplatin, Ronidamin, Lovastatin, Masopropyl, Melalsoprole, Metoclopramide, Mifepristone, Miltefosine, Millimostim, Mispaired double-stranded RNA, Mitoguazone, Mitractol, Mitoxantrone, Morglamostim, Nafarelin, Naloxone + Pentazocine Naltgrastim, Nedaplatin, Niltamide, Noscapine, Novel Erythropoiesis-Promoting Protein, NSC631570 Octreotide, Oprelbequin, Osateron, Oxaliplatin, Paclitaxel, Pamidronic Acid, Pegaspargase, Peginterferon Alpha-2b, Pentosan, Sodium Polysulfate, Pentostatin, Picibanil, Pirarubicin, Rabbit Antithymocyte Polyclonal Antibody, Polyethylene Glycol Interferon Alpha-2a, Porfimer Sodium, Raloxifene, Larcitrexed, Rasbrien Body Me Rasburiembodiment, rhenium etidronate Re186, RII retinamide, rituximab, romultide, samarium (153Sm) lexidonam, salglamostim, schizophyllan, sobuzoxane, sonelmin, strontium-89 chloride, suramin, tasonelmin, tazarotene, tegafur, temoporfin, temozolomide, teniposide, tetrachlorodecaoxide, thalidomide, thymalfacin, thyroid-stimulating hormone alpha, topotecan, toremifene, tositumomab-iodine-131, trastuzumab, treosul Fan, tretinoin, trilostane, trimethrexate, triptorelin, tumor necrosis factor alpha, natural type, ubenimex, bladder cancer vaccine, Maruyama vaccine, melanoma lysate vaccine, barrubicin, verteporfin, vinorelbine, bilirulysine, dinostatin stimulamer or zoledronic acid; Abarelix; AE941 (Aeterna), ambamustin, antisense oligonucleotide, bcl-2 (Genta), APC8015 (Dendreon), decitabine, dexaaminoglutethimide, diazicon, EL532 (Elan),EM800 (Endorecherche), eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant, gallocitabine, gastrin-17 immunogene, HLA-B7 gene therapy (Vical), granulocyte-macrophage colony-stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, ilomastat, IM862 (Cytran), interleukin-2, iproxyfen, LDI200 (Milkhaus), religistim, lintuzumab, CA125 MAb (Biomira), cancer MAb (Japan Pharmaceutical Development), HER-2 and Fc MAb (Medarex), idiotype 105AD7 MAb (CRC Technology), idiotype CEA MAb (Trilex), LYM-1-iodine-131MAb (Techniclone), polymorphoemic mucin-yttrium-90MAb (Antisoma), marimast, menogalil, mitumomab, motexafine, gadolinium, MX6 (Galderma), nelarabine, noratexed, P30 protein, pegvisomant, pemetrexed, porphyromycin, prinomast, RL0903 (Shire), rubitecan, satoraplatin, sodium phenylacetate, sparphosic acid, SRL172 (SR Pharma), SU5416 (SUGEN), TA077 (Tanabe), tetrathiomolybdate, saliblastin, thrombopoietin, tin ethylethiopurine, tirapazamine, cancer vaccine (Biomira), melanoma vaccine (New York University), melanoma vaccine (Sloan Kettering Examples include the melanoma tumor lysis product vaccine (New York Medical College), the viral melanoma cell lysate vaccine (Royal Newcastle Hospital), or Valspodar.

[0377] Further examples of therapeutic agents that may be used in combination with the compounds of the present invention include ipilimumab (Yervoy®); tremelimumab; galiximab; nivolumab, also known as BMS-936558 (Opdivo®); pembrolizumab (Keytruda®); avelumab (Bavencio®); AMP224; BMS-936559; MPDL3280A (also known as RG7446); MEDI-570; AMG557; MGA271; IMP321; BMS-663513; PF-05082566; CDX-1127; anti-OX40 (Providence Health Services)huMAbOX40L; Atasicept; CP-870893; Lucatumumab; Dasetuzumab; Muromonab-CD3; Ipirumumab; MEDI4736 (Imfinzi (registered trademark)); MSB0010718C; AMP 224; adalimumab (Humira®); ado-trastuzumab emtansine (Kadcyla®); aflibercept (Eylea®); alemtuzumab (Campath®); basiliximab (Simulect®); belimumab (Benlysta®); basiliximab (Simulect®); belimumab (Benlysta®); brentuximab vedotin (Adcetris®); canakinumab (Ilaris®); certolizumab pegol (Cimzia®); daclizumab (Zenapax®); daratumumab (Darzalex®) (Registered Trademark)); Denosumab (Prolia(Registered Trademark)); Eculizumab (Soliris(Registered Trademark)); Efalizumab (Raptiva(Trademark)); Gemtuzumab Ozogamicin (Mylotarg(Registered Trademark)); Golimumab (Simponi(Registered Trademark)); Ibritumomab Chiuxetan (Zevalin(Registered Trademark)); Infliximab (Remicade(Registered Trademark)); Motavizumab (Numax(Registered Trademark)); Natalizumab (Tysabri(Registered Trademark)); Obinutuzumab (Gazyva(Registered Trademark)); Ofatumumab (Arzerra(Registered Trademark)); Omalizumab (Xolair(Registered Trademark)); Palivizumab (Synagis(Registered Trademark));Examples include pertuzumab (Perjeta®); pertuzumab (Perjeta®); ranibizumab (Lucentis®); laxibamumab (Abthrax®); tocilizumab (Actemra®); tositumomab; tositumomab-i-131; tositumomab and tositumomab-i-131 (Bexxar®); ustekinumab (Stelara®); AMG 102; AMG 386; AMG 479; AMG 655; AMG 706; AMG 745; and AMG 951.

[0378] The compounds described herein can be used in combination with other agents disclosed herein or other suitable agents, depending on the medical condition being treated. Therefore, in some embodiments, one or more compounds of this disclosure will be co-administered with other therapies described herein. When used in combination therapy, the compounds described herein may be administered simultaneously with or separately from a second agent. This combination administration may include simultaneous administration of the two agents in the same dosage form, simultaneous administration in different dosage forms, and separate administration. That is, the compounds described herein and any of the agents described herein may be formulated together in the same dosage form and administered simultaneously. Alternatively, the compounds of the present invention and any of the therapies described herein may be administered simultaneously, with both agents present in separate formulations. Another alternative is that the compounds of this disclosure may be administered followed by any of the therapies described herein, or vice versa. In some embodiments of separate administration protocols, the compounds of the present invention and any of the therapies described herein may be administered at intervals of minutes, hours, or days.

[0379] In some embodiments of the methods described herein, the first therapy (e.g., the compounds of the present invention) and one or more additional therapies are administered simultaneously or sequentially in any order. The first therapeutic agent may be administered immediately before or after one or more additional therapies, or up to 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, or up to 1-7, 1-14, 1-21, or 1-30 days before or after.

[0380] The present invention also deals with a kit comprising (a) a pharmaceutical composition comprising an agent described herein (e.g., a compound of the present invention), and (b) a package insert containing instructions for carrying out any of the methods described herein. In some embodiments, the kit comprises (a) a pharmaceutical composition comprising an agent described herein (e.g., a compound of the present invention), (b) one or more additional therapies (e.g., non-pharmacological treatments or therapeutic agents), and (c) a package insert containing instructions for carrying out any of the methods described herein.

[0381] One aspect of the present invention aims to treat a disease or related symptoms by a combination of compounds that are pharmaceutically active and can be administered separately. Therefore, the present invention further relates to combining separate pharmaceutical compositions in the form of a kit. The kit may comprise two separate pharmaceutical compositions, namely the compounds of the present invention, and one or more additional therapies. The kit may include containers for housing the separate compositions, such as divided bottles or divided foil packets. Additional examples of containers include syringes, boxes, and bags. In some embodiments, the kit may include instructions for using the separate components. The kit form is particularly advantageous when the separate components are preferably administered in different dosage forms (e.g., orally and parenterally), at different dosing intervals, or when dosage setting of the individual components of the combination is desired by a prescribing healthcare professional. [Examples]

[0382] This disclosure is further illustrated by the following examples and synthesis examples, which should not be construed as limiting the scope or spirit of this disclosure to the specific procedures described herein. It should be understood that these examples are provided to illustrate certain embodiments and are not intended to limit the scope of this disclosure in any way. It should also be understood that various other embodiments, modifications, and equivalents thereof, which may be suggested to those skilled in the art, may be used without departing from the spirit of this disclosure or the appended claims.

[0383] chemical synthesis The following examples and definitions used elsewhere in this specification are set forth below.

[0384] [Table 3]

[0385] device Mass spectrometry data were acquired using a Shimadzu LCMS-2020, Agilent 1260LC-6120 / 6125MSD, Shimadzu LCMS-2010EV, or Waters Acquity UPLC equipped with either a QDa or SQ detector 2. Samples were injected in liquid phase into a C-18 reversed phase. Compounds were eluted from the column using an acetonitrile gradient and fed to the mass spectrometer. Initial data analysis was performed using either an Agilent ChemStation, Shimadzu LabSolutions, or Waters MassLynx. NMR data were acquired using either a Bruker AVANCE III HD 400MHz, Bruker Ascend 500MHz instrument, or Varian 400MHz instrument, and raw data were analyzed using either a TopSpin or Mestrelab Mnova instrument.

[0386] Synthesis of intermediates Synthesis of intermediate 1. (2S)-2-{1-[(2S,3S)-1-(tert-butoxycarbonyl)-2-{[(4-methylbenzenesulfonyl)oxy]methyl}pyrrolidine-3-yl]-N-methylformamide}-3-methylbutanoic acid

[0387] [ka]

[0388] Step 1. To a stirred solution of (benzyloxy)acetic acid (200 g, 1200 mmol) in DCM (2 L), CDI (254 g, 1560 mmol) was added at 0°C. The resulting mixture was stirred at room temperature for 1 hour. Then, TEA (335 mL, 2410 mmol) and N,O-dimethylhydroxylamine hydrochloride (164 g, 1690 mmol) were added at 0°C, and the reaction mixture was stirred at room temperature for 1 hour. The resulting mixture was treated with 1 M aqueous HCl (4 × 800 mL) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 2-(benzyloxy)-N-methoxy-N-methylacetamide (238 g, yield 85%) as a pale yellow oil. LCMS (ESI) m / z: [M + H] C 11 H 15 Calculated value for NO3: 210.1; Measured value: 210.1. Step 2. To a solution of dibenzylamine (177 g, 896 mmol) in DMF (500 mL) stirred at room temperature, tert-butyl 4-bromobutanoate (200 g, 896 mmol), K2CO3 (248 g, 1800 mmol), and KI (14.9 g, 90.0 mmol) were added. The resulting mixture was stirred at 80°C for 2 hours. The reaction reaction was then stopped with H2O at room temperature, extracted with RINKAN (3 × 500 mL), treated with brine, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain tert-butyl 4-(dibenzylamino)butanoate (233 g, yield 71%) as a colorless oil. LCMS (ESI) m / z: [M + H]C 11 H 15 Calculated value for NO3: 340.2; Measured value: 340.2. Step 3. To a stirred solution of DIPA (116 mL, 821 mmol) in anhydrous THF (1.5 L), n-BuLi (328 mL, 821 mmol) was added dropwise at -78°C under an N2 atmosphere. The resulting mixture was stirred at 0°C for 30 minutes and then cooled again to -78°C. Next, tert-butyl 4-(dibenzylamino)butanoate (186 g, 547 mmol) in anhydrous THF (743 mL) was added dropwise, and the resulting mixture was stirred at -78°C for 1 hour. Next, 2-(benzyloxy)-N-methoxy-N-methylacetamide (137 g, 657 mmol) in anhydrous THF (550 mL) was added dropwise, and the resulting mixture was stirred at -78°C for 1 hour. The reaction was then stopped with saturated NH4Cl aqueous solution (500 mL), extracted with toluene (3 × 2 L), dried over anhydrous sodium 2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain tert-butyl 4-(benzyloxy)-2-[2-(dibenzylamino)ethyl]-3-oxobutanoate (168 g, yield 57%) as a yellow oil. LCMS (ESI) m / z: [M + H]C 31 H 37 Calculated value for NO4: 488.3; Measured value: 488.3. Step 4. To a solution of tert-butyl 4-(benzyloxy)-2-[2-(dibenzylamino)ethyl]-3-oxobutanoate (168 g, 344 mmol) and Boc2O (90.1 g, 413 mmol) in THF (1.7 L), 10% Pd / C (83.9 g, 788 mmol) was added. The mixture was hydrogenated under H2 at 2.5 atm at room temperature for 18 hours, filtered through a Celite pad, and concentrated under reduced pressure. The crude product, (cis)-1,3-di-tert-butyl 2-[(benzyloxy)methyl]pyrrolidine-1,3-dicarboxylate (209 g, crude), was used directly in the next step without further purification. LCMS (ESI) m / z: [M + H]C 22 H 33 Calculated value for NO5: 392.2; Measured value: 392.2. Step 5. To a stirred mixture of (cis)-1,3-di-tert-butyl 2-[(benzyloxy)methyl]pyrrolidine-1,3-dicarboxylate (209 g, crude) in DMF (2 L), DBU (407 g, 2670 mmol) was added, and the resulting mixture was stirred overnight at 100°C. The reaction was stopped with H2O at room temperature, extracted with SiO (3 × 1 L), treated with brine (3 × 500 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain (trans)-1,3-di-tert-butyl 2-[(benzyloxy)methyl]pyrrolidine-1,3-dicarboxylate (109 g, 81% yield over two steps) as a pale yellow oil. LCMS (ESI) m / z: [M + H]C 22 H 33 Calculated value for NO5: 392.2; Measured value: 392.2. Step 6. To a solution of (trans)-1,3-di-tert-butyl-2-[(benzyloxy)methyl]pyrrolidine-1,3-dicarboxylate (90 g, 230 mmol) in DCM (750 mL), TFA (250 mL) was added at 0°C. The reaction mixture was stirred overnight at room temperature. The mixture was then concentrated under reduced pressure, and the residue was dissolved in a mixture of THF and H2O. To this mixture, NaHCO3 (96.6 g, 1150 mmol), followed by Boc2O (100 g, 460 mmol) was added at 0°C. The resulting mixture was stirred at room temperature for 1 hour, and then concentrated under reduced pressure. The resulting aqueous mixture was washed with hexane (2 × 200 mL), acidified to pH=6 with concentrated HCl, extracted with a 3:1 volume mixture of DCM / i-PrOH (3 × 300 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to obtain (trans)-2-[(benzyloxy)methyl]-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid (109 g, crude) as a pale red oil. LCMS (ESI) m / z: [M + H]C 18 H 25 Calculated value for NO5: 336.2; Measured value: 336.1. Step 7. To a stirred mixture of (trans)-2-[(benzyloxy)methyl]-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid (109 g, crude) and methyl(2S)-3-methyl-2-(methylamino)butanoate HCl (118 g, 650 mmol) in DMF (1 L), DIPEA (283 mL, 1630 mmol) and HATU (247 g, 650 mmol) were added at 0°C. The resulting mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain tert-butyltrans-2-[(benzyloxy)methyl]-3-{[(2S)-1-methoxy-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}pyrrolidine-1-carboxylate (74 g, 70% yield over two steps) as a red oil. LCMS (ESI) m / z: [M + H] C 25 H 38 Calculated value for N2O6: 463.3; Measured value: 463.0. Step 8. Tert-butyltrans-2-[(benzyloxy)methyl]-3-{[(2S)-1-methoxy-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}pyrrolidine-1-carboxylate (30.0 g, 65.0 mmol) was purified by preparative SFC to obtain tert-butyl(2S,3S)-2-[(benzyloxy)methyl]-3-{[(2S)-1-methoxy-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}pyrrolidine-1-carboxylate (13.0 g, yield 43%) as a yellow oil. LCMS (ESI) m / z: [M + H - 100] C 25 H 38 Calculated value for N2O6: 363.3; Measured value: 363.0. Step 9. A mixture of tert-butyl(2S,3S)-2-[(benzyloxy)methyl]-3-{[(2S)-1-methoxy-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}pyrrolidine-1-carboxylate (12.0 g, 25.9 mmol) and 10% Pd / C (12.0 g, 113 mmol) in a 9:1 volume mixture of MeOH / AcOH (100 mL) was stirred overnight at room temperature under an atmosphere of H2. The reaction mixture was concentrated under reduced pressure to remove MeOH, and the resulting mixture was basicized to pH=8 with saturated NaHCO3 aqueous solution and then extracted with ELISA. The combined organic extracts were washed with saturated NaHCO3 aqueous solution (3 × 100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl(2S,3S)-2-(hydroxymethyl)-3-{[(2S)-1-methoxy-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}pyrrolidine-1-carboxylate (8.53 g, crude). LCMS (ESI) m / z: [M + H - Boc] C 18 H 32 Calculated value for N2O6: 272.2; Measured value: 272.9. Step 10. To a stirred solution of tert-butyl(2S,3S)-2-(hydroxymethyl)-3-{[(2S)-1-methoxy-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}pyrrolidine-1-carboxylate (8.53 g, crude), TEA (19.0 mL, 137 mmol), and DMAP (279 mg, 2.28 mmol) in DCM, a solution of TsCl (19.6 g, 103 mmol) in DCM was added dropwise at 0°C. The resulting mixture was stirred overnight at room temperature. The reaction was stopped by adding H2O (50 mL) at 0°C, then extracted with DCM (3 × 100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain tert-butyl(2S,3S)-3-{[(2S)-1-methoxy-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}-2-{[(4-methylbenzenesulfonyl)oxy]methyl}pyrrolidine-1-carboxylate (9.35 g, 69% yield over two steps) as a yellow solid. LCMS (ESI) m / z: [M + NH4]C 25 H 38 Calculated value for N2O8: 544.3; Measured value: 544.0. Step 11. To a stirred solution of tert-butyl(2S,3S)-3-{[(2S)-1-methoxy-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}-2-{[(4-methylbenzenesulfonyl)oxy]methyl}pyrrolidine-1-carboxylate (9.30 g, 17.7 mmol) in THF (75 mL), a solution of LiOH·H2O (2.22 g, 53.0 mmol) in H2O (15 mL) was added dropwise at 0°C. The resulting mixture was stirred overnight at room temperature. Next, the reaction mixture was concentrated under reduced pressure to remove THF, diluted with H2O (10 mL), acidified to pH=6 with aqueous HCl, extracted with a 3:1 volume mixture of DCM / i-PrOH (3 × 10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain (2S)-2-{1-[(2S,3S)-1-(tert-butoxycarbonyl)-2-{[(4-methylbenzenesulfonyl)oxy]methyl}pyrrolidine-3-yl]-N-methylformamide}-3-methylbutanoic acid (9.62 g, crude), which was used directly in the next step without further purification. LCMS (ESI) m / z: [M + NH4] C 24 H 36 Calculated value for N2O8: 530.3; Measured value: 530.3. Intermediate 1. Alternative synthesis via chiral cleavage

[0389] [ka]

[0390] Step 1. To a stirred solution of (cis)-1,3-di-tert-butyl 2-[(benzyloxy)methyl]pyrrolidine-1,3-dicarboxylate (130 g, 332 mmol) in THF (570 mL), MeOH (260 mL) and LiOH·H2O (41.8 g, 996 mmol) in H2O (260 mL) were added at room temperature under an N2 atmosphere. The resulting mixture was stirred at 40°C for 24 hours. Four of these reactions were carried out in parallel on the same scale. These four reaction mixtures were combined, extracted with petroleum ether (2 × 3 L), washed with 1 M LiOH aqueous solution (2 × 2 L), acidified to pH=3 with 1 M HCl aqueous solution, extracted with SiO2 (3 × 2 L), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain 2-[(benzyloxy)methyl]-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid (414 g, yield 93%) as a white solid. LCMS (ESI) m / z: [M + H]C 18 H 25 Calculated value for NO5: 336.2; Measured value: 336.2. Step 2. To a stirred solution of 2-[(benzyloxy)methyl]-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid (207 g, 617 mmol) in MeOH (2 L), 10 wt% Pd / C (80.1 g) was added in small amounts at room temperature. The resulting mixture was stirred at room temperature for 16 hours under an atmosphere of H2 (10 atm). These reactions were carried out in parallel on the same scale twice. The two resulting mixtures were combined and then filtered. The filtrate was washed with MeOH (3 × 1 L), and the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash column chromatography to obtain 1-(tert-butoxycarbonyl)-2-(hydroxymethyl)pyrrolidine-3-carboxylic acid (266 g, yield 88%) as a white solid. LCMS (ESI) m / z: [2M + Na]C 11 H 19 Calculated value for NO5: 513.2; Measured value: 513.3. Step 3. To a stirred mixture of 1-(tert-butoxycarbonyl)-2-(hydroxymethyl)pyrrolidine-3-carboxylic acid (26.6 g, 108 mmol) in toluene (1460 mL), (R)-[(2S,4R,5S)-5-ethenyl-1-azabicyclo[2.2.2]octan-2-yl](6-methoxyquinoline-4-yl)methanol (35.2 g, 108 mmol) was added in small amounts at room temperature under an atmosphere of N2. The resulting mixture was stirred at room temperature for 5 minutes. The mixture was then heated to 140°C, stirred under reflux for 1 minute, then cooled to 25°C and stirred for 14 hours. Ten of these reactions were carried out in parallel on the same scale. The ten resulting mixtures were then combined and filtered. The filtrate was concentrated under reduced pressure, dissolved in THF (1 L), basicized to pH=8 with 1 N NaOH aqueous solution, extracted with n-hexane (1 × 1 L), and further extracted with ELISA (3 × 500 mL). The aqueous mixture was acidified to pH=3 with 0.5 M HCl and extracted with ELISA (10 × 2 L). The combined organic extracts were dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to obtain (2R,3R)-1-(tert-butoxycarbonyl)-2-(hydroxymethyl)pyrrolidine-3-carboxylic acid (120 g, enantiomer excess >99%, yield 45%) as a colorless oil. LCMS (ESI) m / z: [2M + Na] C 11 H 19 Calculated value for NO5: 513.2; Measured value: 513.2. The remaining filtration cake was washed with i-PrOAc (130 mL) and then diluted with ice water (300 mL). The resulting aqueous mixture was treated with 0.5 M aqueous HCl (700 mL), basicized to pH 8 with 1 N aqueous NaOH (1000 mL), and extracted with i-PrOAc (3 × 700 mL). The remaining aqueous mixture was acidified to pH 3 with 0.5 M aqueous HCl, treated with NaCl (500 g), extracted with ELISA (10 × 2 L), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain (2S,3S)-1-(tert-butoxycarbonyl)-2-(hydroxymethyl)pyrrolidine-3-carboxylic acid (120 g, enantiomer excess 95.1%, yield 45%) as a colorless oil. LCMS (ESI) m / z: [2M + Na]C 11 H 19 Calculated value for NO5: 513.2; Measured value: 513.2. Step 4. To a stirred mixture of (2S,3S)-1-(tert-butoxycarbonyl)-2-(hydroxymethyl)pyrrolidine-3-carboxylic acid (35.0 g, 143 mmol) and benzyl(2S)-3-methyl-2-(methylamino)butanoate (56.8 g, 257 mmol) in MeCN (500 mL), 2,6-lutidine (153 g, 1430 mmol) and a solution of HATU (81.4 g, 214 mmol) in MeCN (200 mL) and DMF (70 mL) were added under an argon atmosphere at -5°C. The reaction mixture was stirred at 0°C for 2 hours, and then the reaction was stopped with H2O (3 L). The resulting mixture was extracted with ELISA (3 × 1 L), washed with brine (3 × 1 L), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography to obtain tert-butyl(2S,3S)-3-{[(2S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}-2-(hydroxymethyl)pyrrolidine-1-carboxylate (42.0 g, yield 66%) as a pale yellow oil. LCMS (ESI) m / z: [M + Na]C 24 H 36 Calculated value for N2O6: 471.3; Measured value: 471.3. Step 5. To a stirred solution of tert-butyl(2S,3S)-3-{[(2S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}-2-(hydroxymethyl)pyrrolidine-1-carboxylate (73.0 g, 163 mmol) in DCM (600 mL), TEA (82.3 g, 814 mmol), followed by a solution of TsCl (40.3 g, 212 mmol) in DCM (100 mL), was added at -5°C under an argon atmosphere. The reaction mixture was stirred at 25°C for 16 hours, and then stopped with H2O (1 L). The mixture was acidified to pH=6 with 1N aqueous HCl and extracted with DCM (3 × 700 mL). After treatment with brine (3 × 500 mL), it was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain tert-butyl(2S,3S)-3-{[(2S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}-2-{[(4-methylbenzenesulfonyl)oxy]methyl}pyrrolidine-1-carboxylate (81.0 g, yield 78%) as a pale yellow solid. LCMS (ESI) m / z: [M + Na]C 31 H 42 Calculated value for N2O8S: 625.3; Measured value: 625.3. Step 6. To a stirred solution of tert-butyl(2S,3S)-3-{[(2S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}-2-{[(4-methylbenzenesulfonyl)oxy]methyl}pyrrolidine-1-carboxylate (87.0 g, 144 mmol) in THF (870 mL), 10 wt% Pd / C (40 g) was added at room temperature. The reaction mixture was stirred under an H2 atmosphere at 25°C for 16 hours. The resulting mixture was then filtered, and the filter cake was washed with MeOH (3 × 500 mL). The filtrate was concentrated under reduced pressure to obtain (2S)-2-{1-[(2S,3S)-1-(tert-butoxycarbonyl)-2-{[(4-methylbenzenesulfonyl)oxy]methyl}pyrrolidine-3-yl]-N-methylformamide}-3-methylbutanoic acid (71.0 g, 96% yield) as a grayish-white solid. LCMS (ESI) m / z: [2M + H]C 24 H 36 Calculated value for N2O8S: 1025.5; Measured value: 1025.3. Intermediate 1. Alternative synthesis via the use of chiral pool starting materials

[0391] [ka]

[0392] Step 1. In a solution of N-(tert-butyloxycarbonyl)-L-aspartic acid β-benzyl ester (4.50 kg, 13.9 mol) in DCM (22.5 L), stirred at 0°C under an N2 atmosphere, DIPEA (2.16 kg, 16.7 mmol) was added, followed by TfOMe (2.74 kg, 16.7 mmol). The resulting mixture was stirred at 20°C for 1 hour. The reaction was stopped by adding 0.5 M aqueous HCl (11.2 L), and then washed with H2O (3 × 11.2 L). The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting concentrated solution was treated with petroleum ether, stirred at 20°C for 30 minutes, and filtered. The filtered cake was washed with petroleum ether and then dried under reduced pressure to obtain 4-(phenylmethyl)-N-[(1,1-dimethylethoxy)carbonyl]-L-methyl aspartate (4.76 kg, crude) as a white solid. LCMS (ESI) m / z: [M + Na]C 17 H 23 Calculated value for NO6: 360.1; Measured value: 359.9. Step 2. Under an N2 atmosphere, a solution of 1.50 kg of 1-methyl 4-(phenylmethyl)-N-[(1,1-dimethylethoxy)carbonyl]-L-aspartate (crude) in THF (12 L) stirred at -75°C was mixed with KHMDS (11.1 L, 1 M in THF), followed by HMPA (1.20 kg, 6.67 mol), and then 3-bromopropa-1-ene (807 g, 6.67 mol). The reaction mixture was stirred at -75°C for 2 hours. The mixture was then diluted with RINKAN, treated with 30% aqueous citric acid solution, and stirred at 20°C for 30 minutes. The aqueous layer was further extracted with RINKAN, treated with brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography to obtain 4-(phenylmethyl)-(3S)-N-[(1,1-dimethylethoxy)carbonyl]-3-(2-propen-1-yl)-L-methyl aspartate (908 g, purity 85.1%, yield 47% over two steps) as a brown oil. LCMS (ESI) m / z: [M + H]C 20 H 27Calculated value for NO6: 378.2; Measured value: 378.2. Step 3. Under an N2 atmosphere, a solution of 1-methyl 4-(phenylmethyl)-(3S)-N-[(1,1-dimethylethoxy)carbonyl]-3-(2-propen-1-yl)-L-aspartate (2.00 kg, 5.30 mol) in MeOH (20 L) and H2O (8 L), stirred at 15°C, was mixed with K2OsO4·2H2O (19.5 g, 53.0 mmol) and NaIO4 (3.40 kg, 15.9 mol). The reaction mixture was stirred at room temperature for 12 hours and then filtered. The filtered cake was washed with MeOH, the filtrate was stopped with 1 M aqueous HCl, treated with saturated aqueous Na2SO3, and concentrated under reduced pressure to remove MeOH. The resulting mixture was extracted with dimethyl sulfate, dried over anhydrous sodium 2SO4, filtered, and concentrated under reduced pressure to obtain 3-benzyl-1-(tert-butyl)-2-methyl-(2S,3S)-5-hydroxypyrrolidine-1,2,3-tricarboxylate (1.79 kg, crude) as a brown oil. LCMS (ESI) m / z: [2M + Na] C 19 H 25 Calculated value for NO7: 781.3; Measured value: 781.4. Step 4. To a solution of 3-benzyl-1-(tert-butyl)-2-methyl-(2S,3S)-5-hydroxypyrrolidine-1,2,3-tricarboxylate (2.00 kg, crude) in DCM (20 L) stirred at room temperature, Et3SiH (919 g, 7.91 mol) was added under an N2 atmosphere. Then TFA (1.80 kg, 15.8 mol) was added dropwise at 15°C, and the reaction mixture was stirred at room temperature for 3 hours. The reaction was then stopped by dropwise addition of saturated NaHCO3 aqueous solution at 15°C, the resulting organic phase was washed with H2O, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was then purified by silica gel chromatography to obtain 3-benzyl-1-(tert-butyl)-2-methyl-(2S,3S)-pyrrolidine-1,2,3-tricarboxylate (945g, purity 69.4%, yield 44% over two steps) as a yellow oil. LCMS (ESI) m / z: [2M + Na]C 19 H 25Calculated value for NO6: 749.3; Measured value: 749.3. Step 5. To a mixture of 20% Pd(OH)2 (12.0 g) in MeOH (500 mL), a solution of 3-benzyl-1-(tert-butyl)-2-methyl-(2S,3S)-pyrrolidine-1,2,3-tricarboxylate (120 g, 330 mmol) in MeOH (500 mL) was added under an argon atmosphere. The reaction mixture was then placed under 45 psi of H2 and stirred at room temperature for 16 hours. The mixture was then filtered through diatomaceous earth, and the filtrate was rinsed with MeOH. The filtrate was concentrated under reduced pressure to obtain (2S,3S)-1-(tert-butoxycarbonyl)-2-(methoxycarbonyl)pyrrolidine-3-carboxylic acid (90 g, purity 78.0%, yield 78%) as a yellow oil. LCMS (ESI) m / z: [M - Boc + 2H] C 12 H 19 Calculated value for NO6: 174.1; Measured value: 174.1. Step 6. To a solution of (2S,3S)-1-(tert-butoxycarbonyl)-2-(methoxycarbonyl)pyrrolidine-3-carboxylic acid (180 g, 659 mmol) in THF (1.8 L) stirred at 0°C under an N2 atmosphere, LiBH4 (2 M in THF, 494 mL) was added. The reaction mixture was stirred at 50°C for 13 hours. The mixture was then cooled to 0°C, the reaction was stopped with H2O, the pH was adjusted to 4 by adding 1 M aqueous HCl, and the mixture was diluted with brine. The aqueous layer was further extracted with ELISA, and the combined organic extract was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain (2S,3S)-1-(tert-butoxycarbonyl)-2-(hydroxymethyl)pyrrolidine-3-carboxylic acid (135 g, crude) as a white solid. This substance was proceeded directly to the next step without further purification.

[0393] Step 7. To a solution of (2S,3S)-1-(tert-butoxycarbonyl)-2-(hydroxymethyl)pyrrolidine-3-carboxylic acid (285 g, 1.16 mol) in MeCN (2.3 L) and DMF (0.3 L), N-methyl-L-valinebenzyl ester HCl salt (119 g, 0.465 mol) was added. This mixture was cooled to 0°C under an N2 atmosphere, and 2,6-lutidine (622 g, 5.81 mol), followed by HATU (442 g, 1.16 mol). The reaction mixture was stirred at 0°C for 2 hours. The reaction was stopped by adding brine, and the organic extract was further treated with brine. The combined aqueous solution was extracted with phenylethylamine, concentrated under reduced pressure, and purified by reverse-phase flash column chromatography to obtain tert-butyl(2S,3S)-3-{[(2S)-1-methoxy-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}-2-(hydroxymethyl)pyrrolidine-1-carboxylate (120 g, 19% yield over two steps) as a white solid. LCMS (ESI) m / z: [M + Na]C 24 H 36 Calculated value for N2O6: 471.3; Measured value: 471.2. Step 8. To a stirred solution of (2S,3S)-3-{[(2S)-1-methoxy-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}-2-(hydroxymethyl)pyrrolidine-1-carboxylate (168 g, 375 mmol) in DCM (1.7 L), TEA (190 g, 1870 mmol) was added, followed by the dropwise addition of a solution of TsCl (92.8 g, 487 mmol) in DCM (1.7 L) under an Ar gas atmosphere at -5°C. The reaction mixture was stirred at room temperature for 16 hours. The resulting mixture was then diluted with water, acidified to pH=6 with 1 M aqueous HCl, extracted with DCM, treated with brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain tert-butyl(2S,3S)-3-{[(2S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}-2-{[(4-methylbenzenesulfonyl)oxy]methyl}pyrrolidine-1-carboxylate (216 g, purity 93%, yield 89%) as a pale yellow solid. LCMS (ESI) m / z: [M + Na]C 31 H 42 Calculated value for N2O5S: 625.3; Measured value: 625.2. Step 9. To a stirred solution of tert-butyl(2S,3S)-3-{[(2S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl](methyl)carbamoyl}-2-{[(4-methylbenzenesulfonyl)oxy]methyl}pyrrolidine-1-carboxylate (216 g, 93% purity, 333 mmol) in THF (2 L), 10 wt% Pd / C (100 g) was added in small amounts at room temperature. The resulting mixture was stirred at room temperature under an H2 gas atmosphere for 16 hours. The resulting mixture was filtered, the filtrate was washed with MeOH, and the filtrate was concentrated under reduced pressure to obtain (2S)-2-{1-[(2S,3S)-1-(tert-butoxycarbonyl)-2-{[(4-methylbenzenesulfonyl)oxy]methyl}pyrrolidine-3-yl]-N-methylformamide}-3-methylbutanoic acid (170 g, purity 91.6%, yield 91%) as a grayish-white solid. LCMS (ESI) m / z: [M + Na]C 31 H42 Calculated value for N2O5S: 535.2; Measured value: 535.3. Synthesis of intermediate 2. tert-butyl(2S,3S)-2-ethynyl-3-(methyl((S)-3-methyl-1-oxo-1-(2-(trimethylsilyl)ethoxy)butan-2-yl)carbamoyl)pyrrolidine-1-carboxylate

[0394] [ka]

[0395] Step 1. To a solution of tert-butyl(2S,3S)-2-((benzyloxy)methyl)-3-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)pyrrolidine-1-carboxylate (9.0 g, 19 mmol) in MeOH (81 mL) and acetic acid (9.0 mL) at room temperature, Pd / C (9 g) was added. The resulting mixture was stirred overnight under an atmosphere of H2, filtered, and the filtrate was washed with MeOH (3 × 100 mL). The filtrate was concentrated under reduced pressure to obtain tert-butyl(2S,3S)-2-(hydroxymethyl)-3-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)pyrrolidine-1-carboxylate (6.8 g, crude), which was then used directly in the next reaction without further purification. LCMS (ESI) m / z: [M + Na] C 18 H 32 Calculated value for N2O6: 395.2; Measured value: 395.2. Step 2: To a solution of tert-butyl(2S,3S)-2-(hydroxymethyl)-3-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)pyrrolidine-1-carboxylate (6.8 g, crude) in MeCN (70 mL) at 0°C, Des Martin periodinane (17.8 g, 41.9 mmol) was added. The resulting mixture was stirred at 0°C for 4 hours, and then the reaction was stopped by adding an aqueous solution of Na2S2O3. The aqueous mixture was extracted with DCM (3 × 100 mL), the combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl(2S,3S)-2-formyl-3-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)pyrrolidine-1-carboxylate (4.0 g, crude), which was used in the next reaction without further purification. LCMS (ESI) m / z: [M + H - 100] C 18 H 30 Calculated value for N2O6: 271.2; Measured value: 271.1. Step 3. Dimethyl (1-diazo-2-oxopropyl)phosphonate (1.97 g, 10.3 mmol) was added to a solution of tert-butyl(2S,3S)-2-formyl-3-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)pyrrolidine-1-carboxylate (3.8 g, crude) and K2CO3 (1.70 g, 12.3 mmol) in MeOH (40 mL) at 0 °C. The resulting mixture was stirred at 0 °C for 3 hours, and then the reaction was stopped by adding H2O (50 mL). The aqueous mixture was extracted with ELISA (3 × 50 mL), and the combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. Reverse-phase chromatography purification yielded tert-butyl(2S,3S)-2-ethynyl-3-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)pyrrolidine-1-carboxylate (2.44 g, 33% yield over 3 steps) as a yellow solid. LCMS (ESI) m / z: [2M + NH4]C 19 H 30Calculated value for N2O5: 750.4; Measured value: 750.5. Step 4. To a solution of tert-butyl(2S,3S)-2-ethynyl-3-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)pyrrolidine-1-carboxylate (2.40 g, 6.55 mmol) in THF (12 mL) and H2O (12 mL) at 0°C, LiOH·H2O (550 mg, 13.1 mmol) was added. The resulting mixture was stirred at room temperature for 2 hours, and then acidified to pH=5 with 1 M aqueous HCl. The aqueous mixture was extracted with toluene (3 × 20 mL), the combined organic extract was dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain N-((2S,3S)-1-(tert-butoxycarbonyl)-2-ethynylpyrrolidine-3-carbonyl)-N-methyl-L-valine (2.0 g, crude), which was used in the next reaction without further purification. LCMS (ESI) m / z: [M + H] C 18 H 28 Calculated value for N2O5: 353.2; Measured value: 353.1. Step 5. To a solution of N-((2S,3S)-1-(tert-butoxycarbonyl)-2-ethynylpyrrolidine-3-carbonyl)-N-methyl-L-valine (1.98 g, crude) and 2-(trimethylsilyl)ethanol (1.33 g, 11.2 mmol) in DCM (20 mL) at 0 °C, EDCI (2.15 g, 11.2 mmol) and DMAP (690 mg, 5.61 mmol) were added. The resulting mixture was stirred at room temperature for 1 hour, and then the reaction was stopped at 0 °C by adding H2O. The aqueous mixture was extracted with DCM (3 × 30 mL), and the combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. Reverse-phase chromatography purification yielded tert-butyl(2S,3S)-2-ethynyl-3-(methyl((S)-3-methyl-1-oxo-1-(2-(trimethylsilyl)ethoxy)butan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (1.95 g, 45% yield over two steps) as a yellow oil. LCMS (ESI) m / z: [M + H]C 23 H 40Calculated value for N2O5Si: 453.3; Measured value: 453.2. Synthesis of intermediate 3. N-((2R,3S)-1-(tert-butoxycarbonyl)-2-vinylpyrrolidine-3-carbonyl)-N-methyl-L-valine

[0396] [ka]

[0397] Step 1. To a solution of tert-butyl(2S,3S)-2-ethynyl-3-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)pyrrolidine-1-carboxylate (7.0 g, 19.1 mmol) and pyridine (1.51 g, 19.1 mmol) in toluene (140 mL), Lindler catalyst (7.0 g) was added. The resulting mixture was stirred at room temperature under an H2 atmosphere for 2 hours, filtered, and the filtrate was washed with MeOH (10 × 10 mL). The filtrate was concentrated under reduced pressure to obtain tert-butyl(2R,3S)-3-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-2-vinylpyrrolidine-1-carboxylate (6.87 g, crude), which was used in the next reaction without further purification. LCMS (ESI) m / z: [M + NH4] C 19 H 32 Calculated value for N2O5: 368.2; Measured value: 368.3. Step 2. To a solution of tert-butyl(2R,3S)-3-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-2-vinylpyrrolidine-1-carboxylate (2.35 g, 6.38 mmol) in THF (30 mL) and H2O (10 mL), LiOH·H2O (540 mg, 12.8 mmol) was added. The resulting mixture was stirred at 0°C for 3 hours, concentrated under reduced pressure, and the concentrate was acidified to pH=5 with 1 M aqueous HCl. The aqueous mixture was extracted with DCM (3 × 15 mL), the combined organic extract was dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain N-((2R,3S)-1-(tert-butoxycarbonyl)-2-vinylpyrrolidine-3-carbonyl)-N-methyl-L-valine (2.36 g, crude), which was used in subsequent reactions without further purification. LCMS (ESI) m / z: [M + Na] C 18 H 30 Calculated value for N2O5: 377.2; Measured value: 377.2. Intermediate 4. tert-butyl (1 2 S,1 3 S, 6S, 9S)-4 5 -Iodo-9-isopropyl-6-((S)-3-(methoxycarbonyl)hexahydropyridazine-1-carbonyl)-10-methyl-8,11-dioxo-3-oxa-7,10-diaza-1(2,3)-pyrrolidina-4(1,3)-benzeneacycloundecafane-1 1 - Synthesis of carboxylates

[0398] [ka]

[0399] Step 1: To a solution of methyl(S)-2-((tert-butoxycarbonyl)amino)-3-(3-iodo-5-((triisopropylsilyl)oxy)phenyl)propanoate (5.0 g, 8.7 mmol) in DMF (50 mL) at 0 °C, CsF (6.58 g, 43.3 mmol) was added. The resulting mixture was stirred at room temperature for 2 hours and the reaction was stopped at 0 °C by adding H2O (50 mL). The aqueous mixture was extracted with RINKAN (3 × 100 mL), the combined organic extract was washed with brine (3 × 100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain methyl(S)-2-((tert-butoxycarbonyl)amino)-3-(3-hydroxy-5-iodophenyl)propanoate (5.02 g, crude), which was used without further purification. LCMS (ESI) m / z: [M + H] C 15 H 20 Calculated value for INO5: 322.0; Measured value: 322.0. Step 2: To a solution of methyl(S)-2-((tert-butoxycarbonyl)amino)-3-(3-hydroxy-5-iodophenyl)propanoate (4.97 g, crude) in DCM (50 mL) at 0°C, TFA (10 mL) was added. The resulting mixture was stirred at room temperature for 1 hour and basicized to pH 8 by adding saturated NaHCO3 aqueous solution. The aqueous layer was extracted with ELISA (3 × 100 mL), and the combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain methyl(S)-2-amino-3-(3-hydroxy-5-iodophenyl)propanoate (4.01 g, crude), which was used without further purification. LCMS (ESI) m / z: [M + H] C 10 H 12 Calculated value for INO3: 322.0; Measured value: 322.0. Step 3: To a solution of methyl(S)-2-amino-3-(3-hydroxy-5-iodophenyl)propanoate (2.0 g, 6.2 mmol) in DMF (20 mL) at 0 °C, DIPEA (43.4 mL, 249 mmol), N-((2S,3S)-1-(tert-butoxycarbonyl)-2-((tosyloxy)methyl)pyrrolidine-3-carbonyl)-N-methyl-L-valine (4.15 g, 8.10 mmol) and COMU (3.47 g, 8.10 mmol) were added. The resulting mixture was stirred at -10 °C for 1 hour, the reaction was stopped at -10 °C by adding H2O (20 mL), and the mixture was neutralized by adding 1 M aqueous HCl. The aqueous mixture was extracted with SiO2 (3 × 50 mL), the combined organic extract was washed with brine (3 × 100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification by normal-phase chromatography yielded tert-butyl(2S,3S)-3-(((S)-1-(((S)-3-(3-hydroxy-5-iodophenyl)-1-methoxy-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-2-((tosyloxy)methyl)pyrrolidine-1-carboxylate (2.60 g, 74% yield over 3 steps) as a yellow solid. LCMS (ESI) m / z: [M + NH4]C 34 H 46 IN3O 10 Calculated value for S: 833.2; Measured value: 833.2. Step 4. To a solution of tert-butyl(2S,3S)-3-(((S)-1-(((S)-3-(3-hydroxy-5-iodophenyl)-1-methoxy-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-2-((tosyloxymethyl)pyrrolidine-1-carboxylate (1.83 g, 2.24 mmol) in DMF (183 mL), K2CO3 (3.10 g, 22.4 mmol) and KI (372 mg, 2.24 mmol) were added. The resulting mixture was stirred at 80°C for 2 hours, and the reaction was stopped at 0°C by adding H2O (100 mL). The aqueous mixture was extracted with dimethyl glycol (3 × 200 mL), the combined organic extract was washed with brine (3 × 400 mL), dried with Na₂SO₄, filtered, and concentrated under reduced pressure. Purification by normal-phase chromatography yielded 1 1 -(tert-butyl)6-methyl(1 2 S,1 3 S, 6S, 9S)-4 5 -Iodo-9-isopropyl-10-methyl-8,11-dioxo-3-oxa-7,10-diaza-1(2,3)-pyrrolidin-4(1,3)-benzeneacycloundecafane-1 1 ,6-dicarboxylate (1.04 g, 66% yield) was obtained as a yellow solid. LCMS (ESI) m / z: [M + NH4]C 27 H 38 Calculated value for IN3O7: 661.2; Measured value: 661.1. Step 5.0℃ THF (6.0mL) 1 -(tert-butyl)6-methyl(1 2 S,1 3 S, 6S, 9S)-4 5 -Iodo-9-isopropyl-10-methyl-8,11-dioxo-3-oxa-7,10-diaza-1(2,3)-pyrrolidin-4(1,3)-benzeneacycloundecafane-1 1A solution of ,6-dicarboxylate (1.04 g, 1.61 mmol) was mixed with a solution of LiOH·H2O (135 mg, 3.22 mmol) in H2O (6.0 mL). The resulting mixture was stirred at 0°C for 1.5 hours and acidified to pH 6 by adding 1 M aqueous HCl. The aqueous mixture was extracted with siRNA (3 × 50 mL), and the combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain (1 2 S,1 3 S, 6S, 9S)-1 1 -(tert-butoxycarbonyl)-4 5 -Iodo-9-isopropyl-10-methyl-8,11-dioxo-3-oxa-7,10-diaza-1(2,3)-pyrrolidin-4(1,3)-benzeneacycloundecafane-6-carboxylic acid (1.27 g, crude) was obtained and used without further purification. LCMS (ESI) m / z: [M + NH4]C 26 H 36 Calculated value for IN3O7: 647.2; Measured value: 647.1. Step 6. In 15 mL of DMF at 0°C (1 2 S,1 3 S, 6S, 9S)-1 1 -(tert-butoxycarbonyl)-4 5 To a solution of -iodo-9-isopropyl-10-methyl-8,11-dioxo-3-oxa-7,10-diaza-1(2,3)-pyrrolidina-4(1,3)-benzeneacycloundecafane-6-carboxylic acid (1.15 g, crude), DIPEA (3.2 mL, 18.3 mmol), methyl(S)-hexahydropyridazine-3-carboxylate bis(trifluoroacetate) (1.36 g, 3.65 mmol), and HATU (1.39 g, 3.65 mmol) were added. The resulting mixture was stirred at room temperature for 1 hour, the reaction was stopped at 0°C by adding H2O (20 mL), and the pH was acidified to 6 by adding 1 M aqueous HCl. The aqueous mixture was extracted with ELISA (3 × 20 mL), the combined organic extract was washed with brine (3 × 30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification by normal-phase chromatography yielded tert-butyl(1 2 S,13 S, 6S, 9S)-4 5 -Iodo-9-isopropyl-6-((S)-3-(methoxycarbonyl)hexahydropyridazine-1-carbonyl)-10-methyl-8,11-dioxo-3-oxa-7,10-diaza-1(2,3)-pyrrolidina-4(1,3)-benzeneacycloundecafane-1 1 -Carboxylates (1.10 g, 72% yield over two steps) were obtained as a pale yellow solid. LC-MS (ESI) m / z: [M + NH4]C 32 H 46 Calculated value for IN5O8: 773.3; Measured value: 773.3. Intermediate 5. tert-butyl((6 3 S,4S)-1 2 -(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-1 1 -ethyl-10,10-dimethyl-5,7-dioxo-2 5 -((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 Synthesis of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate

[0400] [ka]

[0401] Step 1. To a solution of benzyl(S)-4-(5-(5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indole-2-yl)-6-(1-methoxyethyl)pyridine-3-yl)piperazine-1-carboxylate (60.0 g, 90.4 mmol) and methyl(S)-2-((tert-butoxycarbonyl)amino)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-((triisopropylsilyl)oxy)phenyl)propanoate (78.3 g, 136 mmol), K3PO4 (57.6 g, 271 mmol) and Pd(dppf)Cl2·CH2Cl2 (7.36 g, 9.04 mmol) were added. The mixture was stirred overnight at 70°C under a nitrogen atmosphere, and then extracted with ELISA (3 × 200 mL). The organic extract was dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain benzyl 4-(5-(5-(3-((S)-2-((tert-butoxycarbonyl)amino)-3-methoxy-3-oxopropyl)-5-((triisopropylsilyl)oxy)phenyl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indole-2-yl)-6-((S)-1-methoxyethyl)pyridine-3-yl)piperazine-1-carboxylate (92.8 g, 90%) as a yellow solid. LCMS (ESI) m / z: [M + H] C 59 H 83 Calculated value for N5O9Si: 1034.6; Measured value: 1034.7. Step 2. A solution of benzyl 4-(5-(5-(3-((S)-2-((tert-butoxycarbonyl)amino)-3-methoxy-3-oxopropyl)-5-((triisopropylsilyl)oxy)phenyl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indole-2-yl)-6-((S)-1-methoxyethyl)pyridine-3-yl)piperazine-1-carboxylate (92.8 g, 89.7 mmol) and LiOH (3.87 g, 161 mmol) in H2O (1000 mL) and THF (1000 mL) was stirred overnight at 0°C. The resulting mixture was concentrated under reduced pressure and acidified to pH=5 with 1 M HCl (aqueous solution). The resulting mixture was extracted with toluene (3 × 1 L), dried over anhydrous sodium 2SO4, filtered, and concentrated under reduced pressure to obtain (2S)-3-{3-[(2M)-2-(5-{4-[(benzyloxy)carbonyl]piperazin-1-yl}-2-[(1S)-1-methoxyethyl]pyridine-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)indole-5-yl]-5-[(triisopropylsilyl)oxy]phenyl}-2-[(tert-butoxycarbonyl)amino]propanoic acid (92.8 g, crude) as a yellow solid. LCMS (ESI) m / z: [M + H]C 58 H 81 Calculated value for N5O9Si: 1020.6; Measured value: 1020.7. Step 3. To a stirred solution of (2S)-3-{3-[(2M)-2-(5-{4-[(benzyloxy)carbonyl]piperazin-1-yl}-2-[(1S)-1-methoxyethyl]pyridine-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)indole-5-yl]-5-[(triisopropylsilyl)oxy]phenyl}-2-[(tert-butoxycarbonyl)amino]propanoic acid (91.8 g, 90.0 mmol), DIPEA (116 g, 900 mmol), and methyl(S)-hexahydropyridazine-3-carboxylate (19.5 g, 135 mmol) in DMF (1000 mL), HATU (68.4 g, 180 mmol) was added in small amounts at 0°C. The resulting mixture was stirred at room temperature for 2 hours, then diluted with deionized H2O (2 L). The resulting mixture was extracted with ELISA (3 × 2 L), washed with brine (3 × 4 L), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain methyl(S)-1-((S)-3-(3-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indole-5-yl)-5-((triisopropylsilyl)oxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (94.6 g, 92%) as a yellow solid. LCMS (ESI) m / z: [M + H] C 64 H 91 N7O 10 Calculated value for Si: 1146.7; Measured value: 1146.9. Step 4. The solution of (S)-1-((S)-3-(3-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indole-5-yl)-5-((triisopropylsilyl)oxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (110 g, 95.9 mmol) and LiOH (2.76 g, 115 mmol) in THF (900 mL) and H2O (300 mL) was stirred overnight at 0°C. The resulting mixture was concentrated under reduced pressure, acidified to pH=5 with 1 M HCl (aqueous solution), and extracted with ELISA (3 × 500 mL). The combined organic extracts were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain (S)-1-((S)-3-(3-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indole-5-yl)-5-((triisopropylsilyl)oxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid (106 g, crude) as a yellow solid. LCMS (ESI) m / z: [M + H] C 63 H 89 N7O 10 Calculated value for Si: 1132.6; Measured value: 1132.7. Step 5. To a stirred solution of (S)-1-((S)-3-(3-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indole-5-yl)-5-((triisopropylsilyl)oxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid (106 g, 93.2 mmol) and DIPEA (482 g, 3730 mmol) in DCM (10 L), HOBT (126 g, 932 mmol) and EDCI (536 g, 2800 mmol) were added in small amounts at 0°C. The resulting mixture was stirred overnight at room temperature, then concentrated under reduced pressure and diluted with DCM (3 L). The resulting solution was washed with brine, treated with 1 M aqueous HCl (2 × 4 L), neutralized with saturated aqueous NaHCO3 (4 L), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain benzyl 4-(5-((6 3 S,4S)-4-((tert-butoxycarbonyl)amino)-1 1 -ethyl-10,10-dimethyl-5,7-dioxo-2 5 -((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-1(5,3)-Indra-6(1,3)-Pyridadina-2(1,3)-Benzeneacycloundecafane-1 2 (-yl)-6-((S)-1-methoxyethyl)pyridine-3-yl)piperazine-1-carboxylate (68.9 g, 63%) was obtained as a yellow solid. LCMS (ESI) m / z: [M + H]C 63 H 87 Calculated value for N7O9Si: 1114.6; Measured value: 1115.0. Step 6. Benzyl 4-(5-((6) in MeOH (500 mL) 3S,4S)-4-((tert-butoxycarbonyl)amino)-1 1 -ethyl-10,10-dimethyl-5,7-dioxo-2 5 -((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-1(5,3)-Indra-6(1,3)-Pyridadina-2(1,3)-Benzeneacycloundecafane-1 2 A solution of (-yl)-6-((S)-1-methoxyethyl)pyridine-3-yl)piperazine-1-carboxylate (58.7 g, 52.7 mmol) and Pd(OH)2 / C (30.0 g, 214 mmol) was stirred at room temperature under an atmosphere of H2 for 3 hours. The resulting mixture was filtered, and the filtrate was washed with MeOH (3 × 200 mL). The filtrate was then concentrated under reduced pressure to obtain tert-butyl((6 3 S,4S)-1 1 -ethyl-1 2 -(2-((S)-1-methoxyethyl)-5-(piperazin-1-yl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-2 5 -((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (61.4 g, crude) was obtained as a white solid. LCMS (ESI) m / z: [M + H] C 55 H 81 Calculated value for N7O7Si: 980.6; Measured value: 980.7. Step 7. Tert-butyl ((6) in MeOH (250 mL) 3 S,4S)-1 1 -ethyl-1 2-(2-((S)-1-methoxyethyl)-5-(piperazin-1-yl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-2 5 -((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 A solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (25.2 g, 25.7 mmol), (1-ethoxycyclopropoxy)trimethylsilane (11.4 g, 65.5 mmol), and AcOH (3.09 g, 51.4 mmol) was prepared by adding NaBH3CN (3.23 g, 51.4 mmol) at 0°C. The mixture was stirred overnight at 60°C under an N2 atmosphere, basicized to pH=8 with saturated NaHCO3 aqueous solution, and concentrated under reduced pressure. The resulting residue was extracted with ELISA (3 × 150 mL), the combined organic extract was dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain tert-butyl((6) 3 S,4S)-1 2 -(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-1 1 -ethyl-10,10-dimethyl-5,7-dioxo-2 5 -((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (19.3 g, yield 74%) was obtained as a white solid. LCMS (ESI) m / z: [M + H]C 58 H 85 Calculated value for N7O7Si: 1020.6; Measured value: 1021.3. Intermediate 6. tert-butyl(3aS,6S,9S,15S,32aS)-21-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-22-ethyl-6-isopropyl-5,19,19-trimethyl-4,7,10,16-tetraoxo-2,3,3a,4,5,6,7,8,9 Synthesis of 10,13,14,15,16,18,19,20,22,32,32a-Icosahydro-1H,12H-11,15-Epimino-23,25-Etheno-9,28-Methano-26,30-(Metheno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-1-carboxylate

[0402] [ka]

[0403] Step 1. Tert-butyl ((6) in DMF (100 mL) 3 S,4S)-1 2 -(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-1 1 -ethyl-10,10-dimethyl-5,7-dioxo-2 5 -((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 A solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (10.0 g, 9.80 mmol) was treated with CsF (7.44 g, 49,000 mmol) at room temperature for 1 hour. The reaction was stopped with H2O at 0°C. The resulting mixture was then extracted with ELISA (3 × 150 mL), washed with brine (3 × 150 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl((6) 3 S,4S)-1 2-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-1 1 -ethyl-2 5 -Hydroxy-10,10-dimethyl-5,7-dioxo-61,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (8.68 g, crude) was obtained as a yellow solid. LCMS (ESI) m / z: [M + H] C 49 H 65 Calculated value for N7O7: 864.5; Measured value: 854.4. Step 2. tert-butyl((6) in 1,4-dioxane (40 mL) 3 S,4S)-1 2 -(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-1 1 -ethyl-2 5 -Hydroxy-10,10-dimethyl-5,7-dioxo-61,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 To a stirred solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (8.68 g, crude), HCl solution (40 mL, 4.0 M in 1,4-dioxane) was added dropwise at 0°C. The reaction mixture was stirred overnight at room temperature, and then concentrated under reduced pressure to obtain (6 3 S,4S)-4-amino-1 2 -(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-1 1 -Ethyl-25-hydroxy-10,10-dimethyl-6 1 ,6 2 ,6 3,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafane-5,7-dione (9.7 g, crude) was obtained as a yellow solid. LCMS (ESI) m / z: [M + H] C 44 H 57 Calculated value for N7O5: 764.4; Measured value: 764.1. Step 3. (6) of DMF (60 mL) 3 S,4S)-4-amino-1 2 -(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-1 1 -Ethyl-25-hydroxy-10,10-dimethyl-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 To a stirred solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafane-5,7-dione (5.66 g, crude), DIPEA (38.3 g, 296 mmol), N-((2S,3S)-1-(tert-butoxycarbonyl)-2-((tosyloxy)methyl)pyrrolidine-3-carbonyl)-N-methyl-L-valine (3.80 g, 7.41 mmol), and COMU (4.76 g, 11.1 mmol) were added in small amounts at 0°C. The reaction mixture was stirred at 0°C for 1 hour. The reaction was then stopped with H2O at 0°C, the resulting mixture was extracted with ELISA (3 × 100 mL), washed with brine (3 × 300 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain tert-butyl(2S,3S)-3-(((2S)-1-(((6 3 S,4S)-1 2 -(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-1 1-ethyl-2 5 -Hydroxy-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-2-((tosyloxy)methyl)pyrrolidine-1-carboxylate (7.26 g, 99% yield over 3 steps) was obtained as a yellow solid. LCMS (ESI) m / z: [M + H]C 68 H 91 N9O 12 Calculated value for S: 1257.7; Measured value: 1257.7. Step 4. tert-butyl(2S,3S)-3-(((2S)-1-(((6 3 S,4S)-1 2 -(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-1 1 -ethyl-2 5 -Hydroxy-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1To a stirred solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-2-((tosyloxy)methyl)pyrrolidine-1-carboxylate (7.26 g, 5.77 mmol), K2CO3 (7.97 g, 57.7 mmol) and KI (0.96 g, 5.77 mmol) were added in small amounts at 0°C under an atmosphere of N2. The resulting mixture was stirred further at 80°C for 3 hours. The reaction was stopped with H2O at 0°C. The resulting mixture was then extracted with ELISA (3 × 300 mL), treated with brine (3 × 600 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase flash column chromatography to obtain tert-butyl(3aS,6S,9S,15S,32aS)-21-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-22-ethyl-6-isopropyl-5,19,19-trimethyl-4,7,10,16-tetraoxo-2,3,3a,4,5,6,7, 8,9,10,13,14,15,16,18,19,20,22,32,32a-Icosahydro-1H,12H-11,15-Epimino-23,25-Etheno-9,28-Methano-26,30-(Metheno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-1-carboxylate (4.95 g, yield 79%) was obtained as a yellow oil. LCMS (ESI) m / z: [M + H] C 61 H 83 Calculated value for N9O9: 1086.6; measured value: 1086.3. Intermediate 7. tert-butyl((6 3 S,4S)-1 2 -(5-(1-cyclopropylpiperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-25-((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,64 ,6 5 ,6 6 -Hexahydro-1 1 Synthesis of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate

[0404] [ka]

[0405] Step 1. To a solution of benzyl 4-oxopiperidine-1-carboxylate (25 g, 107 mmol) in MeOH (2.5 L) at room temperature, 4-methylbenzene sulfonohydrazide (20 g, 107 mmol) was added. The resulting mixture was stirred at 40°C for 2 hours, and the precipitated solid was collected by filtration to obtain benzyl 4-(2-tosylhydrazineylidene)piperidine-1-carboxylate (30 g, crude), which was used without further purification. LCMS (ESI) m / z: [M + H]C 20 H 23 Calculated value for N3O4S: 402.2; Measured value: 402.2. Step 2. To a solution of (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridine-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-3-yl)-2,2-dimethylpropan-1-ol (20 g, 40.1 mmol) and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (20.3 g, 80.1 mmol), bis(1,5-cyclooctadiene)diiridium(I) dichloride (670 mg, 1.00 mmol) and 4,4'-di-tert-butyl-2,2'-dipyridyl (1.07 g, 4.01 mmol) were added in room temperature THF (80 mL). The resulting mixture was stirred at 55°C for 16 hours, then concentrated under reduced pressure to obtain (S)-3-(5-bromo-2-(2-(1-methoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-3-yl)-2,2-dimethylpropan-1-ol (20.5 g, crude), which was used without further purification. LCMS (ESI) m / z: [M - C6H 10 + H] C 29 H 37 Calculated value for BBrF3N2O4: 543.1; Measured value: 542.9. Step 3. Cs2CO3 (19.9 g, 61.4 mmol) was added to a solution of (S)-3-(5-bromo-2-(2-(1-methoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-3-yl)-2,2-dimethylpropan-1-ol (20.0 g, crude) and benzyl 4-(2-tosylhydrazineylidene)piperidine-1-carboxylate (12.3 g, 30.7 mmol) in dioxane (200 mL) at room temperature. The resulting solution was stirred at 100°C for 10 hours. The reaction product was then filtered, the filter cake was washed with ELISA (3 × 50 mL), and the filtrate was concentrated under reduced pressure. The residue was diluted with H2O (200 mL), and the aqueous layer was extracted with ELISA (2 × 200 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification by normal-phase chromatography yielded benzyl(S)-4-(5-(5-bromo-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indole-2-yl)-6-(1-methoxyethyl)pyridine-3-yl)piperidine-1-carboxylate (15 g, 61% yield over two steps) as a yellow solid. LCMS (ESI) m / z: [M + H]C 36 H 41 Calculated value for BrF3N3O3: 716.2; Measured value: 716.2. Step 4. Benzyl(S)-4-(5-(5-bromo-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indole-2-yl)-6-(1-methoxyethyl)pyridine-3-yl)piperidine-1-carboxylate (15g, 20.9mmol) and (3-((S)-2 To a solution of -((tert-butoxycarbonyl)amino)-3-((S)-3-(methoxycarbonyl)tetrahydropyridazine-1(2H)-yl)-3-oxopropyl)-5-((triisopropylsilyl)oxy)phenyl)boronic acid (12.7 g, 20.9 mmol), K3PO4 (8.89 g, 41.9 mmol) and Pd(dppf)Cl2·DCM (1.71 g, 2.09 mmol) were added. The resulting mixture was stirred at 60°C for 12 hours and diluted at room temperature by adding H2O (200 mL). The aqueous layer was extracted with ELISA (2 × 100 mL), the combined organic extract was dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain methyl(S)-1-((S)-3-(3-(2-(5-(1-((benzyloxy)carbonyl)piperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indole-5-yl)-5-((triisopropylsilyl)oxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (27.6 g, crude), which was used without further purification. LCMS (ESI) m / z: [M + H] C 65 H 89 F3N6O 10 Calculated value for Si: 1199.7; Measured value: 1199.7. Step 5. A solution of methyl(S)-1-((S)-3-(3-(2-(5-(1-((benzyloxy)carbonyl)piperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indole-5-yl)-5-((triisopropylsilyl)oxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (27.0 g, crude) in THF (135 mL) and H2O (135 mL) at 0°C was mixed with LiOH (1.08 g, 45.0 mmol). The resulting mixture was stirred at 0°C for 4 hours and then acidified to pH 6 with 1 M HCl. The aqueous layer was extracted with ELISA (3 × 80 mL), and the combined organic extract was dried over Na2SO4 and concentrated under reduced pressure to obtain (S)-1-((S)-3-(3-(2-(5-(1-((benzyloxy)carbonyl)piperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indole-5-yl)-5-((triisopropylsilyl)oxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid (25.0 g, crude), which was used without further purification. LCMS (ESI) m / z: [M + H] C 64 H 87 F3N6O 10 Calculated value for Si: 1185.6; Measured value: 1185.7. Step 6: To a solution of (S)-1-((S)-3-(3-(2-(5-(1-((benzyloxy)carbonyl)piperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indole-5-yl)-5-((triisopropylsilyl)oxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid (25.0 g, crude) and DIPEA (81.8 g, 632 mmol), EDCI (115 g, 601 mmol) and HOBT (14.3 g, 105 mmol) were added. The obtained solution was stirred overnight at room temperature, concentrated under reduced pressure, and the residue was diluted with saturated NH4Cl aqueous solution (300 mL). The aqueous mixture was extracted with toluene (2 × 300 mL), and the combined organic extract was dried over Na2SO4 and concentrated under reduced pressure. Purification by normal-phase chromatography yielded benzyl 4-(5-((6 3 S,4S)-4-((tert-butoxycarbonyl)amino)-10,10-dimethyl-5,7-dioxo-1 1 -(2,2,2-trifluoroethyl)-2 5 -((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-1(5,3)-Indra-6(1,3)-Pyridadina-2(1,3)-Benzeneacycloundecafane-1 2 (-yl)-6-((S)-1-methoxyethyl)pyridine-3-yl)piperidine-1-carboxylate (10.0 g, 42% yield over 3 steps) was obtained as a yellow oil. LCMS (ESI) m / z: [M + H]C 64 H 85 Calculated value for F3N6O9Si: 1167.6; Measured value: 1167.7. Step 7. Benzyl 4-(5-((6) in IPA (100 mL) 3S,4S)-4-((tert-butoxycarbonyl)amino)-10,10-dimethyl-5,7-dioxo-1 1 -(2,2,2-trifluoroethyl)-2 5 -((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-1(5,3)-Indra-6(1,3)-Pyridadina-2(1,3)-Benzeneacycloundecafane-1 2 To a solution of (10 g, 8.57 mmol) of (-yl)-6-((S)-1-methoxyethyl)pyridine-3-yl)piperidine-1-carboxylate, Pd / C (5.0 g, 10 wt% Pd) was added. The resulting mixture was stirred overnight at room temperature under an atmosphere of H2, filtered, and the filtrate was washed with RINKAN (3 × 50 mL). The filtrate was concentrated under reduced pressure to obtain tert-butyl((6 3 S,4S)-1 2 -(2-((S)-1-methoxyethyl)-5-(piperidine-4-yl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-1 1 -(2,2,2-trifluoroethyl)-2 5 -((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (9.70 g, crude) was obtained and used without further purification. LCMS (ESI) m / z: [M + H] C 56 H 79 Calculated value for F3N6O7Si: 1033.6; Measured value: 1034.7. Step 8.0℃ IPA (93mL) containing tert-butyl ((6 3 S,4S)-1 2-(2-((S)-1-methoxyethyl)-5-(piperidine-4-yl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-1 1 -(2,2,2-trifluoroethyl)-2 5 -((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 To a solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (9.30 g, crude), (1-ethoxycyclopropoxy)trimethylsilane (4.71 g, 27.0 mmol) and AcOH (1.62 g, 27.0 mmol), followed by NaBH3CN (1.70 g, 27.0 mmol), the mixture was stirred overnight at 60°C, and the reaction was stopped by adding saturated NaHCO3 aqueous solution (200 mL). The aqueous mixture was extracted with ELISA (2 × 100 mL), and the combined organic extract was dried over Na2SO4 and concentrated under reduced pressure. Purification by normal-phase chromatography revealed tert-butyl((6 3 S,4S)-1 2 -(5-(1-cyclopropylpiperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-25-((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (5.10 g, 58% yield over two steps) was obtained as a yellow oil. LCMS (ESI) m / z: [M + H]C 56 H 79Calculated value for F3N6O7Si: 1073.6; Measured value: 1074.7. Intermediate 8. tert-butyl(3aS,6S,9S,15S,32aS)-21-(5-(1-cyclopropylpiperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-6-isopropyl-5,19,19-trimethyl-4,7,10,16-tetraoxo-22-(2,2,2-trifluoroethyl)-2,3,3a,4,5, Synthesis of 6,7,8,9,10,13,14,15,16,18,19,20,22,32,32a-Icosahydro-1H,12H-11,15-epimino-23,25-etheno-9,28-methano-26,30-(metheno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-1-carboxylate

[0406] [ka]

[0407] Step 1. Tert-butyl ((6) in DMF (50 mL) 3 S,4S)-1 2 -(5-(1-cyclopropylpiperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-25-((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1To a solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (5.1 g, 4.75 mmol), CsF (3.61 g, 23.8 mmol) was added. The resulting mixture was stirred at room temperature for 1 hour, and the reaction was stopped at 0°C by adding H2O. The aqueous mixture was extracted with ELISA (2 × 50 mL), the combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification by normal-phase chromatography revealed tert-butyl((6 3 S,4S)-1 2 -(5-(1-cyclopropylpiperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-2 5 -Hydroxy-10,10-dimethyl-5,7-dioxo-1 1 -(2,2,2-trifluoroethyl)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (3.20 g, yield 73%) was obtained as a pale yellow solid. LCMS (ESI) m / z: [M + H]C 50 H 63 Calculated value for F3N6O7: 917.5; Measured value: 917.5. Step 2: tert-butyl in dioxane (15 mL) at 0°C ((6 3 S,4S)-1 2 -(5-(1-cyclopropylpiperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-2 5 -Hydroxy-10,10-dimethyl-5,7-dioxo-1 1 -(2,2,2-trifluoroethyl)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6-Hexahydro-1 1 A solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (3.2 g, 3.50 mmol) was mixed with 15 mL of 4 M HCl solution in dioxane. The resulting mixture was stirred at room temperature for 1 hour and then concentrated under reduced pressure. The residue was diluted with H2O (10 mL), and the resulting solution was basicized to pH 8 with saturated NaHCO3 aqueous solution. The aqueous mixture was extracted with DCM (3 × 30 mL), and the combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain (6 3 S,4S)-4-amino-1 2 -(5-(1-cyclopropylpiperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-2 5 -Hydroxy-10,10-dimethyl-1 1 -(2,2,2-trifluoroethyl)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafane-5,7-dione (2.54 g, crude) was obtained and used without further purification. LCMS (ESI) m / z: [M + H] C 45 H 55 Calculated value for F3N6O5: 817.4; Measured value: 817.3. Step 3. (6) in DMF (20 mL) at -10°C 3 S,4S)-4-amino-1 2 -(5-(1-cyclopropylpiperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-2 5 -Hydroxy-10,10-dimethyl-1 1 -(2,2,2-trifluoroethyl)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6-Hexahydro-1 1 To a solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafane-5,7-dione (2.52 g, crude) and N-((2S,3S)-1-(tert-butoxycarbonyl)-2-((tosyloxy)methyl)pyrrolidine-3-carbonyl)-N-methyl-L-valine (2.37 g, 4.62 mmol), DIPEA (21.5 mL, 123 mmol) and COMU (1.85 g, 4.32 mmol) were added. The resulting mixture was stirred at -10°C for 1 hour, and the reaction was stopped at 0°C by adding cold H2O. The aqueous mixture was extracted with ELISA (2 × 10 mL), the combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification by normal-phase chromatography revealed tert-butyl(2S,3S)-3-(((2S)-1-(((6 3 S,4S)-12-(5-(1-cyclopropylpiperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-2 5 -Hydroxy-10,10-dimethyl-5,7-dioxo-1 1 -(2,2,2-trifluoroethyl)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-2-((tosyloxy)methyl)pyrrolidine-1-carboxylate (1.90 g, 41% yield over 2 steps) was obtained as a yellow solid. LCMS (ESI) m / z: [M + H]C 69 H 89 F3N8O 12 Calculated value for S: 1311.6; Measured value: 1311.5. Step 4. Tert-butyl(2S,3S)-3-(((2S)-1-(((6 3S,4S)-12-(5-(1-cyclopropylpiperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-2 5 -Hydroxy-10,10-dimethyl-5,7-dioxo-1 1 -(2,2,2-trifluoroethyl)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 To a solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-2-((tosyloxy)methyl)pyrrolidine-1-carboxylate (1.33 g, 1.01 mmol), K2CO3 (1.40 g, 10.1 mmol) and KI (168 mg, 1.01 mmol) were added. The resulting mixture was stirred at 80°C for 4 hours, and the reaction was stopped at 0°C by adding cold H2O. The aqueous mixture was extracted with ELISA (2 × 100 mL), the combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification by reverse-phase chromatography revealed tert-butyl(3aS,6S,9S,15S,32aS)-21-(5-(1-cyclopropylpiperidine-4-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-6-isopropyl-5,19,19-trimethyl-4,7,10,16-tetraoxo-22-(2,2,2-trifluoroethyl)-2,3,3a,4,5,6,7 ,8,9,10,13,14,15,16,18,19,20,22,32,32a-Icosahydro-1H,12H-11,15-Epimino-23,25-Eteno-9,28-Methano-26,30-(Meteno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-1-carboxylate (600 mg, yield 42%) was obtained as a yellow solid. LCMS (ESI) m / z: [M + H]C 62 H 81Calculated value for F3N8O9: 1139.6; Measured value: 1139.9. Exemplary compound synthesis Examples A25 and A26. (9S,15S,18S,22S)-2-ethyl-18-isopropyl-3-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-5,5,19-trimethyl-8,14,17,20-tetraoxo-2,4,5,6,9,10,11,12,14,15,16,17,18,19,20,21,22,23-octadecahydro-8H-9,13-epimino-1,30-etheno-15,27-methano-25,29-(metheno)pyrrolo[3,4-v][1,18]dioxa[6,9,12]triazacyclotriacontin-22-carb Synthesis of nitriles and (9S,15S,18S,22R)-2-ethyl-18-isopropyl-3-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-5,5,19-trimethyl-8,14,17,20-tetraoxo-2,4,5,6,9,10,11,12,14,15,16,17,18,19,20,21,22,23-octadecahydro-8H-9,13-epimino-1,30-etheno-15,27-methano-25,29-(metheno)pyrrolo[3,4-v][1,18]dioxa[6,9,12]triazacyclotriacontin-22-carboninitrile

[0408] [ka]

[0409] Step 1: To a solution of methyl(S)-3,4-dihydroxybutanoate (1.0 g, 7.5 mmol) and imidazole (1.02 g, 14.9 mmol) in DMF (10 mL) stirred at 0°C, TBDPSCl (2.05 g, 7.46 mmol) was added. The resulting mixture was stirred at room temperature for 3 hours, and the reaction was stopped at 0°C by adding cold H2O. The aqueous mixture was extracted with SiO (3 × 50 mL), the combined organic extract was washed with water, dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification by normal-phase chromatography yielded methyl(S)-4-((tert-butyldiphenylsilyl)oxy)-3-hydroxybutanoate (2.4 g, yield 86%) as a clear oil. LCMS (ESI) m / z: [M + Na]C 21 H 28 Calculated value for O4Si: 395.2; Measured value: 395.2. Step 2: MsCl (461 mg, 4.03 mmol) was added to a solution of methyl(S)-4-((tert-butyldiphenylsilyl)oxy)-3-hydroxybutanoate (1.0 g, 2.7 mmol) and DIPEA (1.04 g, 8.05 mmol) in DCM (20 mL) at 0 °C. The resulting mixture was stirred at room temperature for 1 hour, and the reaction was stopped at 0 °C by adding cold H2O. The aqueous layer was extracted with DCM (2 × 50 mL), the combined organic extract was washed with H2O, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain methyl(S)-4-((tert-butyldiphenylsilyl)oxy)-3-((methylsulfonyl)oxy)butanoate (1.38 g, crude), which was then carried out to the next reaction without further purification. LCMS (ESI) m / z: [M + H] C 22 H 30 Calculated value for O6SSi: 451.2; Measured value: 451.2. Step 3. To a solution of methyl(S)-4-((tert-butyldiphenylsilyl)oxy)-3-((methylsulfonyl)oxy)butanoate (1.38 g, crude) in DMSO (15 mL) stirred at 55°C, NaCN (750 mg, 15.3 mmol) was added. The resulting mixture was stirred at 60°C for 2 hours, and the reaction was stopped at 0°C by adding cold H2O. The aqueous mixture was extracted with RINKAN (2 × 50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification by normal-phase chromatography yielded methyl 4-((tert-butyldiphenylsilyl)oxy)-3-cyanobutanoate (600 mg, 20% yield over two steps) as a brown solid. LCMS (ESI) m / z: [M + H] C 22 H 27 Calculated value for O3Si: 382.2; Measured value: 382.2. Step 4: A solution of methyl 4-((tert-butyldiphenylsilyl)oxy)-3-cyanobutanoate (600 mg, 1.57 mmol) in THF (6.0 mL) and MeOH (6.0 mL) at 0°C was mixed with a solution of LiOH·H2O (99.0 mg, 2.36 mmol) in H2O (6.0 mL). The resulting mixture was stirred at room temperature for 2 hours, then concentrated under reduced pressure to obtain 4-((tert-butyldiphenylsilyl)oxy)-3-cyanobutanoic acid (620 mg, crude), which was used without further purification. LCMS (ESI) m / z: [M + H] C 21 H 25 Calculated value for NO3Si: 368.2; Measured value: 368.2. Step 5: To a solution of methyl N-methyl-L-valinate (558 mg, 3.84 mmol) and Et3N (971 mg, 9.60 mmol) in DMF (10 mL) at 0 °C, 4-((tert-butyldiphenylsilyl)oxy)-3-cyanobutanoic acid (1.18 g, crude) and HATU (1.83 g, 4.80 mmol) were added. The resulting mixture was stirred at room temperature for 1.5 hours, and the reaction was stopped by adding water. The aqueous mixture was extracted with SiO2 (3 × 20 mL), the combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification by normal-phase chromatography yielded methyl N-(4-((tert-butyldiphenylsilyl)oxy)-3-cyanobutanoyl)-N-methyl-L-valinate (770 mg, 99% yield over two steps) as a white solid. LCMS (ESI) m / z: [M + H] C 28 H 38 Calculated value for N2O4Si: 495.3; Measured value: 495.4. Step 6. Trimethylstannanol (292 mg, 1.62 mmol) was added to a solution of methyl N-(4-((tert-butyldiphenylsilyl)oxy)-3-cyanobutanoyl)-N-methyl-L-valine (100 mg, 0.202 mmol) in DCE (2.0 mL) at 80°C. The resulting mixture was stirred at 80°C for 15 hours and concentrated under reduced pressure. Purification by normal-phase preparative TLC yielded N-(4-((tert-butyldiphenylsilyl)oxy)-3-cyanobutanoyl)-N-methyl-L-valine (86 mg, yield 89%) as a white solid. LCMS (ESI) m / z: [M + H]C 27 H 36 Calculated value for N2O4Si: 481.3; Measured value: 481.2. Step 7.0℃ DCM (2.0mL) (6 3 S,4S)-4-amino-1 1 -ethyl-1 2 -(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-10,10-dimethyl-2 5 -((triisopropylsilyl)oxy)-61 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 To a solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafane-5,7-dione (210 mg, 0.235 mmol) and DIPEA (607 mg, 4.70 mmol), N-(4-((tert-butyldiphenylsilyl)oxy)-3-cyanobutanoyl)-N-methyl-L-valine (169 mg, 0.352 mmol) and COMU (151 mg, 0.352 mmol) were added. The resulting mixture was stirred at room temperature for 1 hour, and the reaction was stopped at 0°C by adding cold H2O. The aqueous mixture was extracted with DCM (3 × 5 mL), and the combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification by normal-phase preparative TLC revealed 4-((tert-butyldiphenylsilyl)oxy)-3-cyano-N-((2S)-1-(((6 3 S,4S)-1 1 -ethyl-1 2 -(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-25-((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methylbutanamide (180 mg, yield 57%) was obtained as a white solid. LCMS (ESI) m / z: [M + H]C 78 H 109 Calculated value for N9O8Si2: 1356.8; Measured value: 1357.2. Step 8.0℃ THF (2.0mL) contains 4-((tert-butyldiphenylsilyl)oxy)-3-cyano-N-((2S)-1-(((6 3 S,4S)-1 1 -ethyl-1 2 -(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-25-((triisopropylsilyl)oxy)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 To a solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methylbutanamide (180 mg, 0.133 mmol), TBAF (69.4 mg, 0.266 mmol) was added. The resulting mixture was stirred at room temperature for 2 hours and concentrated under reduced pressure. Purification by normal-phase preparative TLC (20% MeOH / DCM) revealed 3-cyano-N-((2S)-1-(((6 3 S,4S)-1 1 -Ethyl-25-hydroxy-1 2 -(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-4-hydroxy-N-methylbutanamide (117 mg, 92% yield) was obtained as a white solid. LCMS (ESI) m / z: [M + H]C 53 H 71 Calculated value for N9O8: 962.6; Measured value: 962.5. Step 9.0℃ DCM (3.0 mL) contains 3-cyano-N-((2S)-1-(((6 3 S,4S)-1 1 -Ethyl-25-hydroxy-1 2 -(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1DBAD (311 mg, 1.35 mmol) was added to a solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-4-hydroxy-N-methylbutanamide (260 mg, 0.270 mmol) and PPh3 (354 mg, 1.35 mmol). The resulting mixture was stirred at room temperature for 6 hours and then concentrated under reduced pressure. The mixture of the desired product was obtained by normal-phase preparative TLC purification. Next, the diastereomers were separated by reverse-phase chromatography to obtain (9S,15S,18S,22S)-2-ethyl-18-isopropyl-3-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-5,5,19-trimethyl-8,14,17,20-tetraoxo-2,4,5,6,9,10 ,11,12,14,15,16,17,18,19,20,21,22,23-octadecahydro-8H-9,13-epimino-1,30-eteno-15,27-methano-25,29-(meteno)pyrrolo[3,4-v][1,18]dioxa[6,9,12]triazacyclotriacontin-22-carbonitride (11.8mg, yield 4.6%, assumed) (stereoconfiguration) and (9S,15S,18S,22R)-2-ethyl-18-isopropyl-3-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-5,5,19-trimethyl-8,14,17,20-tetraoxo-2,4,5,6,9,10,11,12,14,15,16,17,18, 19,20,21,22,23-octadecahydro-8H-9,13-epimino-1,30-etheno-15,27-methano-25,29-(metheno)pyrrolo[3,4-v][1,18]dioxa[6,9,12]triazacyclotriacontin-22-carbonitrile (13.0 mg, yield 5.1%, assumed stereochemistry) were each obtained as a white solid.

[0410] Data for (9S,15S,18S,22S)-2-ethyl-18-isopropyl-3-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-5,5,19-trimethyl-8,14,17,20-tetraoxo-2,4,5,6,9,10,11,12,14,15,16,17,18,19,20,21,22,23-octadecahydro-8H-9,13-epimino-1,30-etheno-15,27-methano-25,29-(metheno)pyrrolo[3,4-v][1,18]dioxa[6,9,12]triazacyclotriacontin-22-carbonitrile: LCMS (ESI) m / z: [M + H] C 53 H 69 Calculated value for N9O7: 944.5; Measured value: 945.2; s(400 MHz, DMSO-d6) δ 9.33 (s, 1H), 9.28 (s, 1H), 8.72 (d, J = 8.2 Hz, 1H), 8.46 (d, J = 2.9 Hz, 2H), 8.27 (d, J = 7.8 Hz, 1H), 7.91 (d, J = 6.8 Hz, 2H), 7.58 (d, J = 8.6 Hz, 2H), 7.51 (d, J = 8.9 Hz, 2H), 7.33 - 7.21 (m, 6H), 7.18 (d, J = 1.6 Hz, 1H), 7.01 (d, J = 10.6 Hz, 2H), 6.50 (s, 2H), 5.37 (d, J = 11.0 Hz, 4H), 4.67 (d, J = 10.7 Hz, 1H), 4.30 (d, J = 12.0 Hz, 2H), 4.14 - 4.04 (m, 1H), 3.83 (d, J = 10.4 Hz, 2H), 3.65 (d, J = 10.7 Hz, 2H), 3.61 - 3.52 (m, 2H), 3.07 (d, J = 7.7 Hz, 6H), 2.87 (d, J = 14.2 Hz, 1H), 2.77 (d, J = 11.5 Hz, 6H), 2.70 (s, 1H), 2.46 (s, 4H), 2.22 (s, 6H), 2.01 (s, 1H), 1.95 (dd, J = 10.8, 1.5 Hz, 5H), 1.80 (s, 2H), 1.64 (s, 3H), 1.54 (d, J = 12.4 Hz, 2H), 1.35 (d, J = 6.1 Hz, 6H), 1.24 (s, 1H), 0.99 (t, J = 6.8 Hz, 6H), 0.90 (dd, J = 11.6, 6.5 Hz, 5H), 0.80 - 0.70 (m, 10H), 0.56 (d, J = 11.3 Hz, 6H). Data for (9S,15S,18S,22R)-2-ethyl-18-isopropyl-3-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-5,5,19-trimethyl-8,14,17,20-tetraoxo-2,4,5,6,9,10,11,12,14,15,16,17,18,19,20,21,22,23-octadecahydro-8H-9,13-epimino-1,30-etheno-15,27-methano-25,29-(metheno)pyrrolo[3,4-v][1,18]dioxa[6,9,12]triazacyclotriacontin-22-carbonitrile: LCMS (ESI) m / z: [M + H] C 53 H 69 Calculated value for N9O7: 944.5; Measured value: 944.8. 1H NMR (400 MHz, DMSO-d6) δ 9.33 (s, 1H), 9.28 (s, 1H), 8.61 (d, J = 8.4 Hz, 1H), 8.45 (d, J = 2.9 Hz, 2H), 8.12 (d, J = 7.9 Hz, 1H), 7.91 (s, 2H), 7.58 (d, J = 8.6 Hz, 2H), 7.51 (d, J = 8.6 Hz, 2H), 7.33 (s, 1H), 7.24 (s, 3H), 7.17 (s, 1H), 7.01 (s, 2H), 6.50 (s, 2H), 5.35 (s, 5H), 4.69 (d, J = 10.8 Hz, 1H), 4.29 (d, J = 12.9 Hz, 2H), 4.10 (dd, J = 12.4, 6.0 Hz, 2H), 3.90 (d, J = 10.0 Hz, 1H), 3.64 (s, 1H), 3.07 (d, J = 13.4 Hz, 6H), 2.88 (d, J = 13.7 Hz, 1H), 2.81 (s, 6H), 2.46 (s, 5H), 2.21 (d, J = 4.3 Hz, 9H), 2.11 (d, J = 1.7 Hz, 2H), 1.99 (s, 3H), 1.80 (s, 1H), 1.35 (d, J = 6.4 Hz, 5H), 1.24 (s, 3H), 0.98 (s, 1H), 0.99 - 0.88 (m, 5H), 0.81 - 0.74 (m, 10H), 0.58 (s, 2H), 0.53 (s, 3H). Example A43. Synthesis of (9S,15S,18S)-23-acetyl-2-ethyl-18-isopropyl-3-(2-((S)-1-methoxyethyl)pyridine-3-yl)-5,5,19-trimethyl-2,4,5,6,9,10,11,12,15,16,18,19,22,23,24,25-hexadecahydro-8H-9,13-epimino-1,32-etheno-15,29-methano-27,31-(metheno)pyrrolo[3,4-x][1,20]dioxa[4,8,11,14]tetraazacyclodotriacontin-8,14,17,20(21H)-tetraone

[0411] [ka]

[0412] Step 1. To a solution of 2-[(tert-butyldimethylsilyl)oxy]acetaldehyde (2.00 g, 11.5 mmol), tert-butyl 3-aminopropanoate (2.50 g, 17.2 mmol), and DIPEA (2.97 g, 22.9 mmol) in MeOH (20 mL), NaBH3CN (2.16 g, 34.4 mmol) was added at 0°C. The resulting mixture was stirred at room temperature for 5 hours, then extracted with DCM, washed with brine (3 × 50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl 3-({2-[(tert-butyldimethylsilyl)oxy]ethyl}amino)propanoate (2.00 g, crude), which was then carried out without further purification to the next reaction. LCMS (ESI) m / z: [M + H]C 15 H 33 Calculated value for NO3Si: 304.2; Measured value: 304.2. Step 2. To a solution of tert-butyl 3-({2-[(tert-butyldimethylsilyl)oxy]ethyl}amino)propanoate (2.00 g, crude) and DIPEA (1.70 g, 13.2 mmol) in DCM (20 mL), acetyl chloride (0.80 g, 9.88 mmol) was added in small amounts at 0°C. The resulting mixture was stirred at room temperature for 3 hours, and then the reaction was stopped by adding H2O at 0°C. The aqueous layer was extracted with DCM, the combined organic extract was washed with brine (3 × 50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude substance was purified by reverse-phase chromatography to obtain tert-butyl 3-(N-{2-[(tert-butyldimethylsilyl)oxy]ethyl}acetamide)propanoate (2.10 g, 53% yield over two steps) as a yellow oil. LCMS (ESI) m / z: [M + H]C 17 H 35 Calculated value for NO4Si: 346.2; Measured value: 346.3. Step 3. To a solution of tert-butyl 3-(N-{2-[(tert-butyldimethylsilyl)oxy]ethyl}acetamide)propanoate (3.00 g, 8.68 mmol) in DCM (24 mL), TFA (12 mL) was added dropwise at 0°C. The reaction mixture was stirred at room temperature for 1 hour, then neutralized to pH 7 by adding saturated NaHCO3 aqueous solution. The aqueous layer was extracted with DCM, the combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 3-[N-(2-hydroxyethyl)acetamide]propanoic acid (2.2 g, crude), which was used without further purification. LCMS (ESI) m / z: [M + H] C7H 13 Calculated value for NO4: 176.1; Measured value: 176.3. Step 4. Tert-butyl ((6) in MeOH (27 mL) 3 S,4S)-2 5 -(benzyloxy)-1 1 -ethyl-1 2 -(2-((S)-1-methoxyethyl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 To a solution of -hexahydro-11H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (2.70 g, 3.25 mmol), 10% Pd / C (70 mg) was added at room temperature under an N2 atmosphere. The reaction mixture was stirred overnight at room temperature under an H2 atmosphere. The resulting mixture was then filtered, the filtrate was washed with DCM:MeOH by volume 10:1, and the filtrate was concentrated under reduced pressure to obtain tert-butyl((6 3 S,4S)-1 1 -ethyl-2 5 -Hydroxy-1 2 -(2-((S)-1-methoxyethyl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,65 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (2.40 g, crude) was obtained and used without further purification. LCMS (ESI) m / z: [M + H] C 42 H 53 Calculated value for N5O7: 740.4; Measured value: 740.7. Step 5. tert-butyl((6) in 1,4-dioxane (8 mL) 3 S,4S)-1 1 -ethyl-2 5 -Hydroxy-1 2 -(2-((S)-1-methoxyethyl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 A solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)carbamate (1.50 g, crude) was mixed with HCl (8 mL, 4 M solution in 1,4-dioxane) at 0°C. The resulting mixture was stirred at room temperature for 2 hours, and then neutralized to pH 8 by adding saturated NaHCO3 aqueous solution. The aqueous mixture was extracted with ELISA, the combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain (6 3 S,4S)-4-amino-1 1 -ethyl-2 5 -Hydroxy-1 2 -(2-((S)-1-methoxyethyl)pyridine-3-yl)-10,10-dimethyl-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafane-5,7-dione (1.00 g, crude) was obtained and used without further purification. LCMS (ESI) m / z: [M + H] C 37 H 45 Calculated value for N5O4: 640.3; Measured value: 640.3. Step 6. (6) of DMF (10 mL) 3 S,4S)-4-amino-1 1 -ethyl-2 5 -Hydroxy-1 2 -(2-((S)-1-methoxyethyl)pyridine-3-yl)-10,10-dimethyl-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 To a solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafane-5,7-dione (1.00 g, crude) and DIPEA (8.08 g, 62.5 mmol), (2S)-2-[(tert-butoxycarbonyl)(methyl)amino]-3-methylbutanoic acid (0.54 g, 2.35 mmol) and COMU (80.0 g, 1.88 mmol) were added at 0°C. The resulting mixture was stirred under a nitrogen atmosphere at room temperature for 2 hours, and the reaction was stopped by adding H2O. The aqueous mixture was extracted with ELISA, the combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by preparative TLC to obtain tert-butyl((2S)-1-(((6 3 S,4S)-1 1 -ethyl-2 5 -Hydroxy-1 2 -(2-((S)-1-methoxyethyl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-11 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate (800 mg, 46% yield over 3 steps) was obtained as a yellow solid. LCMS (ESI) m / z: [M + H]C 48 H 64 Calculated value for N6O8: 853.5; Measured value: 853.6. Step 7. tert-butyl((2S)-1-(((6) in 1,4-dioxane (2 mL) 3 S,4S)-1 1 -ethyl-2 5 -Hydroxy-1 2 -(2-((S)-1-methoxyethyl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 A solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate (380 mg, 0.445 mmol) was mixed with HCl (2 mL, 4 M solution in 1,4-dioxane) at 0°C. The resulting mixture was stirred at room temperature for 2 hours and then concentrated under reduced pressure. The residue was neutralized to pH 8 by adding saturated NaHCO3 aqueous solution. The aqueous mixture was extracted with ELISA (5 × 100 mL), the combined organic extract was washed with brine (3 × 50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain (2S)-N-((6 3 S,4S)-1 1 -ethyl-2 5 -Hydroxy-1 2 -(2-((S)-1-methoxyethyl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,65 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(1,3)-benzeneacycloundecaphane-4-yl)-3-methyl-2-(methylamino)butanamide (250 mg, crude) was obtained and used without further purification. LCMS (ESI) m / z: [M + H] C 43 H 56 Calculated for C28H43N6O4: 753.4; Found 753.3. Step 8. A solution of (2S)-N-((6 3 S,4S)-1 1 -ethyl-2 5 -hydroxy-1 2 -(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -hexahydro-1 1 H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(1,3)-benzeneacycloundecaphane-4-yl)-3-methyl-2-(methylamino)butanamide (200 mg, crude) and DIPEA (137 g, 1.06 mmol) in DMF (2 mL) was added dropwise with 3-[N-(2-hydroxyethyl)acetamido]propanoic acid (69.8 mg, crude) and HATU (151 mg, 0.399 mmol) at 0 °C. The resulting mixture was stirred at room temperature for 2 h and then quenched by the addition of H2O. The aqueous mixture was extracted with EtOAc, the combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated in vacuo. The crude residue was purified by preparative TLC to give (2S)-N-((6 3 S,4S)-1 1 -ethyl-2 5 -hydroxy-1 2 -(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,63 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-1(5,3)-Indra-6(1,3)-Pyridadina-2(1,3)-Benzeneacycloundecafan-4-yl)-2-(3-(N-(2-hydroxyethyl)acetamide)-N-methylpropanamide)-3-methylbutanamide (60.0 mg, 19% yield over two steps) was obtained as a yellow solid. LCMS (ESI) m / z: [M + H]C 50 H 67 Calculated value for N7O9: 910.5; Measured value: 910.8. Step 9. (2S)-N-((6) in toluene (20 mL) 3 S,4S)-1 1 -ethyl-2 5 -Hydroxy-1 2 -(2-((S)-1-methoxyethyl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1To a solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)-2-(3-(N-(2-hydroxyethyl)acetamide)-N-methylpropanamide)-3-methylbutanamide (35.0 mg, 0.038 mmol), CMBP (46.4 mg, 0.190 mmol) was added at 0°C. The resulting mixture was stirred at room temperature for 2 hours, then stirred overnight at 60°C, and the reaction was stopped by adding H2O. The aqueous mixture was extracted with ELISA, the combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by reverse-phase chromatography to obtain (9S,15S,18S)-23-acetyl-2-ethyl-18-isopropyl-3-(2-((S)-1-methoxyethyl)pyridine-3-yl)-5,5,19-trimethyl-2,4,5,6,9,10,11,12,15,16,18,19,22,23,24,25-hexadecahydro-8H-9,13-epimino-1,32-etheno-15,29-methano-27,31-(metheno)pyrrolo[3,4-x][1,20]dioxa[4,8,11,14]tetraazacyclodotriacontin-8,14,17,20(21H)-tetraone (1.1 mg, yield 3.0%) as a white solid. LCMS (ESI) m / z: [M + H] C 50 H 65 Calculated value for N7O8: 892.5; Measured value: 892.8. 1H NMR (400 MHz, DMSO-d6) δ 8.76 (dd, J = 4.8, 1.7 Hz, 1H), 8.61 - 8.38 (m, 1H), 8.02 (d, J = 5.4 Hz, 1H), 7.89 - 7.72 (m, 2H), 7.60 - 7.48 (m, 2H), 7.35 (s, 1H), 7.28 - 7.14 (m, 1H), 6.77 - 6.72 (m, 1H), 5.35 - 5.10 (m, 2H), 4.80 (d, J = 11.0 Hz, 1H), 4.37 - 3.76 (m, 10H), 3.70 - 3.58 (m, 2H), 3.55 - 3.40 (m, 5H), 3.10 (d, J = 5.9 Hz, 3H), 3.00 - 2.70 (m, 7H), 2.70 - 2.61(m, 1H), 2.21 - 1.95 (m, 6H), 1.85 - 1.61 (m, 2H), 1.59 - 1.51 (m, 1H), 1.41 - 1.34 (m, 4H), 1.24 (br s, 1H), 1.11 - 0.62 (m, 14H), 0.61 - 0.33 (m, 3H). Example A62. (9S,13S,16S,19S,25S)-33-ethyl-16-isopropyl-10-(3-methoxyazetidine-1-carbonyl)-32-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-15,29,29-trimethyl-7,27-dioxa-10,15,18,21,33,39-hexaazaheptacyclo[29.5.2.1 2,6 .1 4,19 .1 21,25 .0 9,13 .0 34,38 Synthesis of hentetraconta-1(37),2(41),3,5,31,34(38),35-heptaene-14,17,20,26-tetraone

[0413] [ka]

[0414] Step 1: (9S,13S,16S,19S,25S)-33-ethyl-16-isopropyl-32-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-15,29,29-trimethyl-7,27-dioxa-10,15,18,21,33,39-hexaazaheptacyclo[29.5.2.1] in 1 mL of DCM at 0°C. 2,6 .1 4,19 .1 21,25 .0 9,13 .0 34,38 To a solution of hentetraconta-1(37),2(41),3,5,31,34(38),35-heptaene-14,17,20,26-tetraone (60.0 mg, 0.696 mmol), DIPEA (90.0 mg, 0.696 mmol), followed by triphosgene (8.26 mg, 0.028 mmol), was added. The resulting mixture was stirred at room temperature for 1 hour, and then 3-methoxyazetidine hydrochloride (7.74 mg, 0.063 mmol) was added at 0°C, and the resulting mixture was stirred at room temperature for 2 hours. The mixture was then filtered and concentrated under reduced pressure, and the resulting crude substance was purified by reverse-phase chromatography to obtain (9S,13S,16S,19S,25S)-33-ethyl-16-isopropyl-10-(3-methoxyazetidine-1-carbonyl)-32-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-15,29,29-trimethyl-7,27-dioxa-10,15,18,21,33,39-hexaazaheptacyclo[29.5.2.1 2,6 .1 4,19 .1 21,25 .0 9,13 .0 34,38 Hentetrakonta-1(37),2(41),3,5,31,34(38),35-heptaene-14,17,20,26-tetraone (40 mg, 57% yield) was obtained as a white solid. LCMS (ESI) m / z: [M + H]C 54 H 70 Calculated value for N8O9: 975.5; Measured value: 975.9; 1H NMR (400 MHz, DMSO-d6) δ = 8.76 (d, J = 4.0 Hz, 1H), 8.23 ​​(d, J = 8.0 Hz, 1H), 8.02 (s, 1H), 7.86 - 7.79 (m, 1H), 7.76 - 7.70 (m, 1H), 7.61 - 7.44 (m, 2H), 7.42 - 7.31 (m, 1H), 7.27 - 7.13 (m, 1H), 6.83 - 6.72 (m, 1H), 5.37 - 5.08 (m, 2H), 4.72 (d, J = 12.0 Hz, 1H), 4.44 - 4.17 (m, 7H), 4.15 - 3.76 (m, 5H), 3.72 - 3.42 (m, 5H), 3.23 (s, 3H), 3.15 - 3.09 (m, 3H), 2.90 - 2.71 (m, 4H), 2.4 (s, 3H), 2.22 - 2.11 (m, 1H), 2.08 - 1.95 (m, 2H), 1.93 - 1.79 (m, 1H), 1.77 - 1.49 (m, 3H), 1.46 -1.36 (m, 3H), 1.30 - 1.12 (m, 1H), 1.00 - 0.64 (m, 12H), 0.50 - 0.44 (m, 3H). Example A80. (3aS,6S,9S,15S,32aS)-21-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-22-ethyl-6-isopropyl-5,19,19-trimethyl-1-propionyl-3,3a,5,6,8,9,14,15,18,19, Synthesis of 20,22,32,32a-tetradecahydro-1H,12H-11,15-epimino-23,25-etheno-9,28-methano-26,30-(metheno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-4,7,10,16(2H,13H)-tetraone

[0415] [ka]

[0416] Step 1. In DCM (20 mL), tert-butyl(3aS,6S,9S,15S,32aS)-21-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-22-ethyl-6-isopropyl-5,19,19-trimethyl-4,7,10,16-tetraoxo-2,3,3a,4,5,6,7,8,9,10,13,14,15,16,18, To a stirred solution of 19,20,22,32,32a-icosahydro-1H,12H-11,15-epimino-23,25-etheno-9,28-methano-26,30-(metheno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-1-carboxylate (4.70 g, 4.33 mmol), TFA (4.00 mL, 35.1 mmol) was added dropwise at 0°C. The reaction mixture was stirred at room temperature for 1 hour, then concentrated under reduced pressure to obtain (3aS,6S,9S,15S,32aS)-21-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-22-ethyl-6-isopropyl-5,19,19-trimethyl-3,3a,5,6,8,9,14,15,18,19 ,20,22,32,32a-tetradecahydro-1H,12H-11,15-epimino-23,25-etheno-9,28-methano-26,30-(metheno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-4,7,10,16(2H,13H)-tetraone (5.96g, crude) was obtained. LCMS (ESI) m / z: [M + H] C 56 H 75 Calculated value for N9O7: 986.6; Measured value: 986.5. Step 2. (3aS,6S,9S,15S,32aS)-21-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-22-ethyl-6-isopropyl-5,19,19-trimethyl-3,3a,5,6,8,9,14,15,18,19,20,22,32,32a-tetradecahydro-1H,12H-11,15-epimino-23,25-eth To a stirred solution of no-9,28-methano-26,30-(metheno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-4,7,10,16(2H,13H)-tetraone (310 mg, crude), DIPEA (406 mg, 3.14 mmol), propanoic acid (46.6 mg, 0.628 mmol), and HATU (239 mg, 0.628 mmol) were added. The reaction mixture was stirred at 0°C for 30 minutes, and then stopped by adding H2O (40 mL) at 0°C. The resulting mixture was extracted with ELISA (3 × 10 mL) and concentrated under reduced pressure. The residue was purified by reverse-phase preparative HPLC to obtain (3aS,6S,9S,15S,32aS)-21-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-22-ethyl-6-isopropyl-5,19,19-trimethyl-1-propionyl-3,3a,5,6,8,9,14,15,18,19,20,22,32, 32a-Tetradecahydro-1H,12H-11,15-epimino-23,25-etheno-9,28-methano-26,30-(metheno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-4,7,10,16(2H,13H)-tetraone (99 mg, 42% yield over two steps) was obtained as a white solid. LCMS (ESI) m / z: [M + H]C 59 H 79 Calculated value for N9O8: 1042.6; Measured value: 1042.5. 1H NMR (400 MHz, DMSO-d6) δ 8.48 (s, 1H), 8.20 - 7.93 (m, 1H), 7.75 - 7.70 (m, 1H), 7.61 - 7.54 (m, 1H), 7.50 - 7.40 (m, 1H), 7.39 - 7.12 (m, 2H), 6.83 - 6.05 (m, 2H), 5.46 - 5.00 (m, 2H), 4.71 - 4.51 (m, 1H), 4.47 (s, 1H), 4.45 - 4.30 (m, 1H), 4.33 - 3.88 (m, 5H), 3.90 - 3.60 (m, 4H), 3.23 (s, 3H), 3.10 - 2.95 (m, 5H), 2.90 - 2.71 (m, 7H), 2.69 - 2.60 (m, 1H), 2.39 (s, 3H), 2.32 - 2.16 (m, 4H), 2.10 - 1.90 (m, 3H), 1.90 - 1.79 (m, 2H), 1.77 - 1.55 (m, 4H), 1.50 - 1.15 (m, 6H), 1.10 - 0.90 (m, 6H), 0.91 - 0.43 (m, 16H). Example A126. (3aS,6S,9S,15S,32aS)-22-ethyl-6-isopropyl-21-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-5,19,19-trimethyl-1-(thiazole-2-yl)-3,3a,5,6,8,9,14,15,18,1 Synthesis of 9,20,22,32,32a-tetradecahydro-1H,12H-11,15-epimino-23,25-etheno-9,28-methano-26,30-(metheno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-4,7,10,16(2H,13H)-tetraone

[0417] [ka]

[0418] Step 1: To a solution of tert-butyl(2S,3S)-2-((benzyloxy)methyl)-3-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)pyrrolidine-1-carboxylate (3.0 g, 6.5 mmol) in DCM (24 mL) at 0°C, TFA (8.0 mL) was added. The resulting mixture was stirred overnight at room temperature and then concentrated under reduced pressure. The residue was dissolved in DCM (10 mL), and the resulting solution was basicized to pH 8 with saturated NaHCO3 aqueous solution. The aqueous layer was extracted with DCM (3 × 10 mL), the combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain N-methyl-N-((2S,3S)-1-(thiazole-2-yl)-2-((tosyloxy)methyl)pyrrolidine-3-carbonyl)-L-valinate (2.86 g, crude), which was used without further purification. LCMS (ESI) m / z: [M + H] C 20 H 30 Calculated value for N2O4: 363.2; Measured value: 362.9. Step 2. To a solution of methyl N-((2S,3S)-2-((benzyloxy)methyl)pyrrolidine-3-carbonyl)-N-methyl-L-valinenate (2.87 g, 7.91 mmol) and 2-bromothiazole (3.89 g, 23.7 mmol) in toluene (30 mL), Cs2CO3 (7.74 g, 23.7 mmol), Pd(OAc)2 (1.07 g, 4.75 mmol), and BINAP (5.91 g, 9.45 mmol) were added. The resulting mixture was stirred overnight at 80°C, and the reaction was stopped at 0°C by adding cold H2O. The aqueous layer was extracted with SiO2 (3 × 10 mL), and the combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. Reverse-phase chromatography purification yielded methyl N-((2S,3S)-2-((benzyloxy)methyl)-1-(thiazole-2-yl)pyrrolidine-3-carbonyl)-N-methyl-L-valinate (923 mg, yield 26%) as a red oil. LCMS (ESI) m / z: [M + H]C 23 H 31 Calculated value for N3O4S: 446.2; Measured value: 446.2. Step 3: To a solution of methyl N-((2S,3S)-2-((benzyloxy)methyl)-1-(thiazole-2-yl)pyrrolidine-3-carbonyl)-N-methyl-L-valinate (821 mg, 1.84 mmol) in DCM (4.0 mL) at 0°C, FeCl3 (1.20 g, 7.38 mmol) was added. The resulting mixture was stirred overnight at room temperature, and the reaction was stopped at 0°C by adding saturated NaHCO3 aqueous solution. The mixture was filtered, and the aqueous layer was extracted with DCM (3 × 5 mL). The filtered cake was washed with DCM (3 × 5 mL), followed by MeOH (3 × 10 mL). The combined organic extract was concentrated under reduced pressure to obtain methyl N-((2S,3S)-2-(hydroxymethyl)-1-(thiazole-2-yl)pyrrolidine-3-carbonyl)-N-methyl-L-valinate (934 mg, crude), which was used without further purification. LCMS (ESI) m / z: [M + H] C 15 H 25 Calculated value for N3O4S: 356.2; Measured value: 356.2. Step 4: To a solution of methyl N-((2S,3S)-2-(hydroxymethyl)-1-(thiazole-2-yl)pyrrolidine-3-carbonyl)-N-methyl-L-valinate (933 mg, crude) and Et3N (1.06 g, 10.5 mmol) in DCM (4.0 mL) at 0 °C, TsCl (1.50 g, 7.88 mmol) and DMAP (32 mg, 0.26 mmol) were added. The resulting mixture was stirred at room temperature for 2 hours, and then the reaction was stopped at 0 °C by adding cold H2O. The aqueous layer was extracted with DCM (3 × 10 mL), and the combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification by normal-phase chromatography yielded methyl N-methyl-N-((2S,3S)-1-(thiazole-2-yl)-2-((tosyloxy)methyl)pyrrolidine-3-carbonyl)-L-valinate (644 mg, yield 67%) as a brown oil. LCMS (ESI) m / z: [M + H]C 23 H 31 Calculated value for N3O6S2: 510.2; Measured value: 510.2. Step 5. To a solution of methyl N-methyl-N-((2S,3S)-1-(thiazol-2-yl)-2-((tosyloxy)methyl)pyrrolidine-3-carbonyl)-L-valinate (320 mg, 0.628 mmol) in THF (5.0 mL) at 0 °C was added LiOH·H2O (79 mg, 1.8 mmol) and H2O (1.0 mL). The resulting mixture was stirred at room temperature overnight, and the reaction solution was acidified to pH 6 with 2 M aqueous HCl. The aqueous mixture was extracted with DCM (3 × 20 mL), and the combined organic extracts were dried over Na2SO4, filtered, and concentrated under reduced pressure to give N-methyl-N-((2S,3S)-1-(thiazol-2-yl)-2-((tosyloxy)methyl)pyrrolidine-3-carbonyl)-L-valine (355 mg, crude), which was used without further purification. LCMS (ESI) m / z: [M + H] C 22 H 29 Calculated for C17H25N3O6S2: 496.2; found 496.1. Step 6. (6 3 S,4S)-4-Amino-1 1 -ethyl-2 5 -hydroxy-1 2 -(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -hexahydro-1 1To a solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-5,7-dione (264 mg, 0.358 mmol) and N-methyl-N-((2S,3S)-1-(thiazole-2-yl)-2-((tosyloxy)methyl)pyrrolidine-3-carbonyl)-L-valine (177 mg, crude), DIPEA (1.85 g, 14.3 mmol) and COMU (230 mg, 0.537 mmol) were added. The resulting mixture was stirred at 0°C for 1.5 hours, and the reaction was stopped at 0°C by adding cold H2O. The aqueous mixture was extracted with ELISA (3 × 10 mL), the combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification by reverse-phase chromatography yields ((2S,3S)-3-(((2S)-1-(((6 3 S,4S)-1 1 -ethyl-2 5 -Hydroxy-1 2 -(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-1-(thiazole-2-yl)pyrrolidine-2-yl)methyl4-methylbenzenesulfonate (187 mg, yield 43%) was obtained as a yellow solid. LCMS (ESI) m / z: [M + H]C 64 H 82 N 10 O 10 Calculated value for S2: 1215.6; Measured value: 1215.2. Step 7. ((2S,3S)-3-(((2S)-1-(((6 3 S,4S)-1 1 -ethyl-25 -Hydroxy-1 2 -(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 To a solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafan-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-1-(thiazole-2-yl)pyrrolidine-2-yl)methyl4-methylbenzenesulfonate (185 mg, 0.152 mmol), K2CO3 (210 mg, 1.52 mmol) and KI (25.3 mg, 0.152 mmol) were added. The resulting mixture was stirred at 80°C for 2 hours, and the reaction was stopped with H2O at -78°C. The aqueous mixture was extracted with ELISA (3 × 20 mL), the combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification by reverse-phase chromatography revealed (3aS,6S,9S,15S,32aS)-22-ethyl-6-isopropyl-21-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazine-1-yl)pyridine-3-yl)-5,19,19-trimethyl-1-(thiazole-2-yl)-3,3a,5,6,8,9,14,15,18,19,20,2 2,32,32a-tetradecahydro-1H,12H-11,15-epimino-23,25-etheno-9,28-methano-26,30-(metheno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-4,7,10,16(2H,13H)-tetraone (24.2 mg, yield 15%) was obtained as a white solid. LCMS (ESI) m / z: [M + H]C 57 H 74 N 10 Calculated value for O7S: 1043.6; Measured value: 1043.4. 1H NMR (400 MHz, DMSO-d6) δ 8.45 (d, J = 2.6 Hz, 1H), 8.39 - 8.17 (m, 1H), 8.11 - 7.95 (m, 1H),7.83 - 7.66 (m, 1H), 7.64 - 7.50 (m, 2H), 7.49 - 7.36 (m, 1H), 7.34 - 7.11 (m, 2H), 7.08 - 6.52 (m, 2H), 5.46 - 5.06 (m, 2H), 4.83 - 4.66 (m, 1H), 4.64 - 4.46 (m, 1H), 4.44 -3.80 (m, 7H), 3.77 - 3.60 (m, 5H), 3.35 - 3.11 (m, 5H), 3.09 - 2.95 (m, 2H), 2.91 -2.57 (m, 5H), 2.53 - 2.35 (m, 6H), 2.33 - 2.12 (m, 4H), 1.92 - 1.51 (m, 6H), 1.45 - 1.15 (m, 4H), 1.09 - 0.93 (m, 2H), 7.83 - 7.66 (m, 1H), 0.91 - 0.51 (m, 12H), 0.49 - 0.37 (m, 1H). Example A130. (3aS,6S,9S,15S,32aS)-21-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-22-ethyl-6-isopropyl-5,19,19-trimethyl-1-(oxetane-3-ylmethyl)-3,3a,5,6,8,9,14,15,1 Synthesis of 8,19,20,22,32,32a-tetradecahydro-1H,12H-11,15-epimino-23,25-etheno-9,28-methano-26,30-(metheno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-4,7,10,16(2H,13H)-tetraone

[0419] [ka]

[0420] Step 1. (3aS,6S,9S,15S,32aS)-21-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-22-ethyl-6-isopropyl-5,19,19-trimethyl-3,3a,5,6,8,9,14,15,18,19,20,22,32,32a-tetradecahydro-1H,12H-11,15-epimino-23,25-etheno-9,28-methano-26,30 -(meteno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-4,7,10,16(2H,13H)-tetraone (200 mg, 0.203 mmol) and oxetane-3-carbaldehyde (69.8 mg, 0.812 mmol) were stirred at 0°C, to which NaBH3CN (51 mg, 0.812 mmol) and ZnCl2 (111 mg, 0.812 mmol) were added, and the mixture was then heated to 60°C. After 1 hour, the reaction was cooled to 0°C, the reaction was stopped with water (5 mL), concentrated under reduced pressure, basicized to pH 8 with saturated NaHCO3 aqueous solution, extracted in siRNA (3 × 10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained residue was purified by reverse-phase column chromatography to obtain (3aS,6S,9S,15S,32aS)-21-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-22-ethyl-6-isopropyl-5,19,19-trimethyl-1-(oxetane-3-ylmethyl)-3,3a,5,6,8,9,14,15,18 ,19,20,22,32,32a-tetradecahydro-1H,12H-11,15-epimino-23,25-etheno-9,28-methano-26,30-(metheno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-4,7,10,16(2H,13H)-tetraone (47.6 mg, yield 22%) was obtained as a pale yellow solid. LCMS (ESI) m / z: [M + H]C 60 H 81 Calculated value for N9O8: 1056.6; Measured value: 1056.4. 1H NMR (400 MHz, DMSO-d6) δ 8.45 (s, 1H), 8.30 - 8.15 (m, 1H), 7.95 (s, 1H), 7.68 (d, J = 8.8 Hz, 1H), 7.56 (d, J = 8.7 Hz, 1H), 7.45 - 7.38 (m, 1H), 7.19 (s, 2H), 6.66 (s, 1H), 5.29 - 5.08 (m, 2H), 4.86 - 4.49 (m, 3H), 4.42 - 4.02 (m, 8H), 3.89 - 3.70 (m, 4H), 3.25 - 3.13 (m, 8H), 3.07 - 2.95 (s, 4H), 2.98 - 2.89 (m, 2H), 2.83 - 2.63 (m, 8H), 2.34 - 2.20 (m, 2H), 2.06 (s, 4H), 1.88 - 1.45 (m, 6H), 1.41 - 1.17 (m, 4H), 1.09 - 0.59 (m, 12H), 0.57 - 0.24 (m, 7H). Example A138. (3aS,6S,9S,15S,32aS)-21-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-22-ethyl-6-isopropyl-5,19,19-trimethyl-1-(methylsulfonyl)-3,3a,5,6,8,9,14,15,18, Synthesis of 19,20,22,32,32a-tetradecahydro-1H,12H-11,15-epimino-23,25-etheno-9,28-methano-26,30-(metheno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-4,7,10,16(2H,13H)-tetraone

[0421] [ka]

[0422] Step 1. In DCM (4 mL), tert-butyl(9S,13S,16S,19S,25S)-32-[5-(4-cyclopropylpiperazine-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridyl]-33-ethyl-16-isopropyl-15,29,29-trimethyl-14,17,20,26-tetraoxo-7,27-dioxa-10,15,18,21,33,39-hexaazaheptacyclo[29.5.2.1 2,6 .1 4,19 .1 21,25 .0 9,13 .0 34,38 To a stirred solution of hentetrakonta-1(37),2(41),3,5,31,34(38),35-heptaene-10-carboxylate (100 mg, 0.101 mmol), TEA (103 mg, 1.01 mmol) and MsCl (23.2 mg, 0.202 mmol) were added at 0°C. The reaction mixture was stirred at room temperature for 30 minutes, and then the reaction was stopped by adding H2O (10 mL) at 0°C. The aqueous phase was extracted with DCM (3 × 10 mL), the combined organic extracts were dried over Na2SO4, filtered, and concentrated under reduced pressure. Next, the residue was purified by reverse-phase chromatography to obtain (3aS,6S,9S,15S,32aS)-21-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-22-ethyl-6-isopropyl-5,19,19-trimethyl-1-(methylsulfonyl)-3,3a,5,6,8,9,14,15,18,19, 20,22,32,32a-tetradecahydro-1H,12H-11,15-epimino-23,25-etheno-9,28-methano-26,30-(metheno)dipyrrolo[2,3-c:3',4'-v][1,18]dioxa[6,9,12]triazacyclotriacontin-4,7,10,16(2H,13H)-tetraone (12.8 mg, yield 12%) was obtained as a white solid. LCMS (ESI) m / z: [M + H]C 57 H 77 Calculated value for N9O9S: 1064.6; Measured value: 1064.4. 1H NMR (300 MHz, DMSO-d6) δ 8.46 (s, 1H), 8.27 (d, J = 8.0 Hz, 1H), 8.00 (s, 1H), 7.73 (d, J = 8.4 Hz, 1H), 7.64 - 7.49 (m, 1H), 7.43 (d, J = 12.3 Hz, 1H), 7.35 - 7.09 (m, 2H), 6.81 - 6.53 (m, 1H), 5.33 (d, J = 12.0 Hz, 1H), 4.84 - 3.98 (m, 7H), 3.69 (s, 4H), 3.23 (s, 6H), 3.06 (d, J = 5.7 Hz, 5H), 2.88 (d, J = 14.2 Hz, 4H), 2.70 (d, J = 11.9 Hz, 7H), 2.23 (d, J = 23.9 Hz, 1H), 2.03 (d, J = 11.9 Hz, 2H), 1.66 - 1.82 (m, 5H), 1.45 - 1.17 (m, 5H), 1.00 (d, J = 7.0 Hz, 2H), 0.95 - 0.65 (m, 10H), 0.64 - 0.20 (m, 8H). Example A140. Synthesis of N-((9S,15S,18S,22S)-3-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-2-ethyl-18-isopropyl-5,5,19-trimethyl-8,14,17,20-tetraoxo-2,4,5,6,9,10,11,12,14,15,16,17,18,19,20,21,22,23-octadecahydro-8H-9,13-epimino-1,30-etheno-15,27-methano-25,29-(metheno)pyrrolo[3,4-v][1,18]dioxa[6,9,12]triazacyclotriacontin-22-yl)-N-methylacetamide

[0423] [ka]

[0424] Step 1: A solution of (S)-4-(benzyloxy)-2-((tert-butoxycarbonyl)amino)-4-oxobutanoic acid (10 g, 30.9 mmol) in THF (100 mL) stirred at 0°C was mixed with BH3·THF (93 mL, 971 mmol). The resulting mixture was stirred at room temperature for 3 hours, the reaction was stopped at 0°C by adding MeOH (10 mL), and the mixture was concentrated under reduced pressure. Purification by normal-phase chromatography yielded benzyl(S)-3-((tert-butoxycarbonyl)amino)-4-hydroxybutanoate (4.69 g, yield 49%) as a yellow solid. LCMS (ESI) m / z: [M + Na + MeCN] C 16 H 23 Calculated value for NO5: 373.2; Measured value: 373.2. Step 2. To a solution of benzyl(S)-3-((tert-butoxycarbonyl)amino)-4-hydroxybutanoate (507 mg, 1.64 mmol) and imidazole (558 mg, 8.20 mmol) in DMF (5.0 mL), TBDPSCl (371 mg, 2.46 mmol) was added. The resulting mixture was stirred at room temperature for 3 hours, and then the reaction was stopped by adding H2O (15 mL). The resulting mixture was extracted with RINKAN (3 × 20 mL), the combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain benzyl(S)-3-((tert-butoxycarbonyl)amino)-4-((tert-butyldiphenylsilyl)oxy)butanoate (448 mg, crude), which was used in the next reaction without further purification. LCMS (ESI) m / z: [M + H] C 32 H 41 Calculated value for NO5Si: 548.3; Measured value: 548.5. Step 3: NaH (131 mg, 5.48 mmol) was added to a solution of benzyl(S)-3-((tert-butoxycarbonyl)amino)-4-((tert-butyldiphenylsilyl)oxy)butanoate (1.5 g, crude) and MeI (583 mg, 4.11 mmol) in DMF (15 mL) stirred at 0 °C. The resulting mixture was stirred overnight at 60 °C, and the reaction was stopped at 0 °C by adding cold H2O. The aqueous mixture was extracted with RINKAN (3 × 50 mL), and the combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by reverse-phase chromatography to obtain benzyl(S)-3-((tert-butoxycarbonyl)(methyl)amino)-4-((tert-butyldiphenylsilyl)oxy)butanoate (753 mg, 25% yield over two steps) as a clear oil. LCMS (ESI) m / z: [M + H] C 33 H 43 Calculated value for NO5Si: 562.3; Measured value: 562.2. Step 4. To a solution of benzyl(S)-3-((tert-butoxycarbonyl)(methyl)amino)-4-((tert-butyldiphenylsilyl)oxy)butanoate (500 mg, 0.890 mmol) in MeOH (10 mL), Pd / C (167 mg) was added. The resulting mixture was stirred at room temperature under an atmosphere of H2 for 1 hour, filtered, and the filtrate was washed with MeOH (3 × 20 mL). The filtrate was concentrated under reduced pressure to obtain (S)-3-((tert-butoxycarbonyl)(methyl)amino)-4-((tert-butyldiphenylsilyl)oxy)butanoic acid (387 mg, crude), which was used in the next reaction without further purification. LCMS (ESI) m / z: [M + H]C 26 H 37 Calculated value for NO5Si: 472.3; Measured value: 472.5. Step 5: To a solution of (S)-3-((tert-butoxycarbonyl)(methyl)amino)-4-((tert-butyldiphenylsilyl)oxy)butanoic acid (590 mg, crude) and benzylmethyl-L-valinate (415 mg, 1.88 mmol) in DMF (12 mL) at 0 °C, Et3N (2.53 g, 25.0 mmol) and HATU (951 mg, 2.50 mmol) were added. The resulting mixture was stirred at 0 °C for 2 hours, and the reaction was stopped by adding H2O. The aqueous mixture was extracted with RINKAN (3 × 30 mL), and the combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification of the crude substance by normal-phase chromatography yielded benzyl N-((S)-3-((tert-butoxycarbonyl)(methyl)amino)-4-((tert-butyldiphenylsilyl)oxy)butanoyl)-N-methyl-L-valinate (776 mg, 85% yield over two steps) as a clear oil. LCMS (ESI) m / z: [M + H]C 39 H 54 Calculated value for N2O6Si: 675.4; Measured value: 675.4. Step 6. To a solution of benzyl N-((S)-3-((tert-butoxycarbonyl)(methyl)amino)-4-((tert-butyldiphenylsilyl)oxy)butanoyl)-N-methyl-L-valine (100 mg, 0.178 mmol) in MeOH (10 mL), Pd / C (100 mg) was added. The resulting mixture was stirred at room temperature under a hydrogen atmosphere for 1 hour, filtered, and the filtrate was washed with MeOH (3 × 10 mL). The filtrate was concentrated under reduced pressure to obtain N-((S)-3-((tert-butoxycarbonyl)(methyl)amino)-4-((tert-butyldiphenylsilyl)oxy)butanoyl)-N-methyl-L-valine (78 mg, crude), which was used in the next reaction without further purification. LCMS (ESI) m / z: [M + H]C 32 H 48 Calculated value for N2O6Si: 585.3; Measured value: 585.4. Step 7.0℃ DCM (10mL) (6 3 S,4S)-4-amino-1 2-(5-(4-cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridine-3-yl)-1 1 -ethyl-2 5 -Hydroxy-10,10-dimethyl-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 To a solution of H-8-oxa-1(5,3)-indra-6(1,3)-pyridadina-2(1,3)-benzeneacycloundecafane-5,7-dione (613 mg, 0.802 mmol) and N-((S)-3-((tert-butoxycarbonyl)(methyl)amino)-4-((tert-buty...

Claims

1. Formula Ia or Formula Ib: 【Chemistry 1】 A compound having the structure thereof, or a pharmaceutically acceptable salt thereof [in the formula, A is a 3- to 6-membered heterocycloalkylene that can be optionally substituted, a 3- to 6-membered cycloalkylene that can be optionally substituted, a 6-membered arylene that can be optionally substituted, or a 5- to 10-membered heteroarylene that can be optionally substituted. L is a linker, R １ These are 5-10 member heteroaryls that are optionally substituted, R ２ C is replaced by any choice. １ ~C ６ It is alkyl, R ３ is optionally substituted C １ to C ６ alkyl, optionally substituted C １ to C ３ heteroalkyl, or optionally substituted 3- to 6-membered cycloalkyl, and R ４ C is substituted with hydrogen or optionally. １ ~C ６ It is alkyl, Each R ３３ These are, independently, halogens, and optionally substituted C １ ~C ３ Alkyl, optionally substituted C １ ~C ３ The alkoxy, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 6-membered heterocycloalkyl, t is 0, 1, 2, or 3. z is 0, 1, or 2. X ９ -NR Ｌ６ -, -C(O)-, or -S(O) ２ - and R Ｌ１ , R Ｌ２ , R Ｌ３ , R Ｌ４ , R Ｌ４ , R Ｌ５ , and R Ｌ６ Each of them can be independently substituted with hydrogen, halogen, hydroxyl, or C of any choice. １ ~C ６ Alkyl, optionally substituted C ２ ~C ６ Alkenyl, C which can be optionally replaced. ２ ~C ６ Alkinyl, or C as optionally replaced. １ ~C ６ It is heteroalkyl, or R Ｌ１ , R Ｌ２ , R Ｌ３ , R Ｌ４ , R Ｌ４ , R Ｌ５ , and R Ｌ６ Any two of these atoms, along with the atoms bonded to them and any intervening atoms, are optionally substituted. ３ ~C ８ [Forms cycloalkyl or 3- to 8-membered heterocyclines.]

2. Equation Ia-1: 【Chemistry 2】 A compound according to claim 1 having the structure, or a pharmaceutically acceptable salt thereof [in which, A is a 3- to 6-membered heterocycloalkylene that can be optionally substituted, a 3- to 6-membered cycloalkylene that can be optionally substituted, a 6-membered arylene that can be optionally substituted, or a 5- to 10-membered heteroarylene that can be optionally substituted. L is a linker, R １ These are 5-10 member heteroaryls that are optionally substituted, R ２ C is replaced by any choice. １ ~C ６ It is alkyl, R ３ C is replaced by any choice. １ ~C ６ Alkyl, optionally substituted C １ ~C ３ A heteroalkyl group, or a 3- to 6-membered cycloalkyl group that is optionally substituted. R ４ C is substituted with hydrogen or optionally. １ ~C ６ It is alkyl, Each R ３３ These are, independently, halogens, and optionally substituted C １ ~C ３ Alkyl, optionally substituted C １ ~C ３ The alkoxy, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 6-membered heterocycloalkyl, t is 0, 1, 2, or 3.

3. Equation Ia-2: 【Transformation 3】 A compound according to claim 2 having the structure, or a pharmaceutically acceptable salt thereof [in which case, A is a 3- to 6-membered heterocycloalkylene that can be optionally substituted, a 3- to 6-membered cycloalkylene that can be optionally substituted, a 6-membered arylene that can be optionally substituted, or a 5- to 10-membered heteroarylene that can be optionally substituted. L is a linker, R １ These are 5-10 member heteroaryls that are optionally substituted, R ２ C is replaced by any choice. １ ~C ６ It is alkyl, R ３ C is replaced by any choice. １ ~C ６ Alkyl, optionally substituted C １ ~C ３ A heteroalkyl group, or a 3- to 6-membered cycloalkyl group that is optionally substituted. R ４ C is substituted with hydrogen or optionally. １ ~C ６ It is alkyl.

4. The aforementioned compound is of formula IIa or formula IIb: 【Chemistry 4】 A compound according to claim 1 having the structure, or a pharmaceutically acceptable salt thereof [in which, R ５ This is hydrogen, optionally substituted 3- to 10-membered heterocycloalkyl groups, -OR ５ａ , or C which is replaced by any choice １ ~C ６ It is heteroalkyl, R ５ａ C is replaced by any choice. １ ~C ６ [It is an alkyl group or a 5- to 10-membered heteroaryl group that is optionally substituted.]

5. The aforementioned compound is of formula IIa-1: 【Transformation 5】 A compound according to claim 4 having the structure, or a pharmaceutically acceptable salt thereof [in which case, R ５ This is hydrogen, optionally substituted 3- to 10-membered heterocycloalkyl groups, -OR ５ａ , or C which is replaced by any choice １ ~C ６ It is heteroalkyl, R ５ａ C is replaced by any choice. １ ~C ６ A 5-10 member heteroaryl that is alkyl or optionally substituted, Each R ３３ is independently halogen, optionally substituted C １ to C ３ alkyl, optionally substituted C １ to C ３ alkoxy, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 6-membered heterocycloalkyl, t is 0, 1, 2, or 3.

6. The aforementioned compound is of formula IIa-2: 【Transformation 6】 A compound according to claim 5 having the structure, or a pharmaceutically acceptable salt thereof [in which case, R ５ This is hydrogen, optionally substituted 3- to 10-membered heterocycloalkyl groups, -OR ５ａ , or C which is replaced by any choice １ ~C ６ It is heteroalkyl, R ５ａ is optionally substituted C １ to C ６ alkyl or optionally substituted 5- to 10-membered heteroaryl].

7. The compound is of formula IIIa or formula IIIb: 【Transformation 7】 A compound according to claim 4 having the structure, or a pharmaceutically acceptable salt thereof.

8. The aforementioned compound is given by formula IIIa-1: 【Transformation 8】 A compound according to claim 7 having the structure, or a pharmaceutically acceptable salt thereof.

9. The aforementioned compound is of formula IIIa-2: 【Chemistry 9】 A compound according to claim 8 having the structure, or a pharmaceutically acceptable salt thereof.

10. L is in formula IV: 【Chemistry 10】 It has a structure, X １ is O or CH ２ It is bonded to ring A, Z is a 3- to 6-membered heterocycloalkylene that can be optionally substituted, and C is an optionally substituted compound. １ ~C ６ Alkylene, or C as optionally substituted. １ ~C ６ A compound according to any one of claims 1 to 9, which is a heteroalkylene, or a pharmaceutically acceptable salt thereof.

11. The aforementioned compound is of formula Va or formula Vb: 【Chemistry 11】 A compound according to claim 10 having the structure, or a pharmaceutically acceptable salt thereof [in which case, X １ is O or CH ２ And, Z is a 3- to 6-membered heterocycloalkylene that can be optionally substituted, and C is an optionally substituted compound. １ ~C ６ Alkylene, or C as optionally substituted. １ ~C ６ It is a heteroalkylene.

12. The aforementioned compound is of formula Va-1: 【Chemistry 12】 A compound according to claim 11 having the structure, or a pharmaceutically acceptable salt thereof [in which case, X １ is O or CH ２ And, Z is a 3- to 6-membered heterocycloalkylene that can be optionally substituted, and C is an optionally substituted compound. １ ~C ６ Alkylene, or C as optionally substituted. １ ~C ６ It is a heteroalkylene.

13. The aforementioned compound is given by formula Va-2: 【Chemistry 13】 A compound according to claim 12 having the structure, or a pharmaceutically acceptable salt thereof [in which, X １ is O or CH ２ And, Z is a 3- to 6-membered heterocycloalkylene that can be optionally substituted, and C is an optionally substituted compound. １ ~C ６ Alkylene, or C as optionally substituted. １ ~C ６ It is a heteroalkylene.

14. L is given by equation VI: 【Chemistry 14】 It has a structure, B is a 3- to 6-membered heterocycloalkylene that is optionally substituted. R ６ C is a hydrogen atom, optionally substituted. １ ~C ６ Alkyl, optionally substituted C １ ~C ６ Heteroalkyls, optionally substituted 3-6 member heterocyclyls, optionally substituted 3-6 member cycloalkyls, optionally substituted 5-10 member heteroaryls, optionally substituted C ６ ~C １０ Ariel, 【Chemistry 15】 And, R ７ and R ８ Each of these can be independently replaced by H or C of any choice. １ ~C ６ It is alkyl, R ９ C is replaced by any choice. １ ~C ６ Alkyl, optionally substituted C ２ ~C ６ The alkenyl, optionally substituted 3-6 membered cycloalkyl, or optionally substituted 3-6 membered heterocyclyl, R １０ C is replaced by any choice. １ ~C ６ Alkyl, optionally substituted C ２ ~C ６ Alkenyl, C which can be optionally replaced. １ ~C ６ Heteroalkyl, optionally substituted C ３ ~C １０ Cycloalkyl, optionally substituted 3-6 membered heterocyclyl, optionally substituted 5-10 membered heteroaryl, or optionally substituted C ６ ~C １０ It is Ariel, R １１ C is a hydrogen atom, optionally substituted. １ ~C ６ Alkyl, optionally substituted C １ ~C ６ Heteroalkyl, optionally substituted C ２ ~C ６ Alkenyl, C which can be optionally replaced. ２ ~C ６ Heteroalkenyl, C which is optionally substituted. ２ ~C ６ Alkinyl, C, which can be optionally replaced. ２ ~C ６ Heteroalkynyl, C which is optionally substituted. ３ ~C １０ Cycloalkyl, optionally substituted 3- to 10-membered heterocyclyl, optionally substituted C ３ ~C １０ Cycloalkenyls, optionally substituted 3- to 10-membered heterocycloalkenyls, optionally substituted C ６ ~C １０ A compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, wherein the compound is an aryl or optionally substituted 5 to 10-membered heteroaryl.

15. L is given by equation VIa: 【Chemistry 16】 A compound according to claim 14 having the structure, or a pharmaceutically acceptable salt thereof.

16. The compound is defined as formula VIIa or formula VIIb: 【Chemistry 17】 A compound according to claim 15 having the structure, or a pharmaceutically acceptable salt thereof [in which, R ６ C is a hydrogen atom, optionally substituted. １ ~C ６ Alkyl, optionally substituted C １ ~C ６ Heteroalkyls, optionally substituted 3-6 member heterocyclyls, optionally substituted 3-6 member cycloalkyls, optionally substituted 5-10 member heteroaryls, optionally substituted C ６ ~C １０ Ariel, [Chemistry 18] And, R ７ and R ８ Each of these can be independently replaced by H or C of any choice. １ ~C ６ It is alkyl, R ９ C is replaced by any choice. １ ~C ６ Alkyl, optionally substituted C ２ ~C ６ The alkenyl, optionally substituted 3-6 membered cycloalkyl, or optionally substituted 3-6 membered heterocyclyl, R １０ C is replaced by any choice. １ ~C ６ Alkyl, optionally substituted C ２ ~C ６ Alkenyl, C which can be optionally replaced. １ ~C ６ Heteroalkyl, optionally substituted C ３ ~C １０ Cycloalkyl, optionally substituted 3-6 membered heterocyclyl, optionally substituted 5-10 membered heteroaryl, or optionally substituted C ６ ~C １０ It is Ariel, R １１ C is a hydrogen atom, optionally substituted. １ ~C ６ Alkyl, optionally substituted C １ ~C ６ Heteroalkyl, optionally substituted C ２ ~C ６ Alkenyl, C which can be optionally replaced. ２ ~C ６ Heteroalkenyl, C which is optionally substituted. ２ ~C ６ Alkinyl, C, which can be optionally replaced. ２ ~C ６ Heteroalkynyl, C which is optionally substituted. ３ ~C １０ Cycloalkyl, optionally substituted 3- to 10-membered heterocyclyl, optionally substituted C ３ ~C １０ Cycloalkenyls, optionally substituted 3- to 10-membered heterocycloalkenyls, optionally substituted C ６ ~C １０ [An aryl, or a 5- to 10-membered heteroaryl that is optionally substituted.]

17. The aforementioned compound is of formula VIIa-1: 【Chemistry 19】 A compound according to claim 16 having the structure, or a pharmaceutically acceptable salt thereof [in which case, R ６ C is a hydrogen atom, optionally substituted. １ ~C ６ Alkyl, optionally substituted C １ ~C ６ Heteroalkyls, optionally substituted 3-6 member heterocyclyls, optionally substituted 3-6 member cycloalkyls, optionally substituted 5-10 member heteroaryls, optionally substituted C ６ ~C １０ Ariel, 【Chemistry 20】 And, R ７ and R ８ Each of these can be independently replaced by H or C of any choice. １ ~C ６ It is alkyl, R ９ C is replaced by any choice. １ ~C ６ Alkyl, optionally substituted C ２ ~C ６ The alkenyl, optionally substituted 3-6 membered cycloalkyl, or optionally substituted 3-6 membered heterocyclyl, R １０ C is replaced by any choice. １ ~C ６ Alkyl, optionally substituted C ２ ~C ６ Alkenyl, C which can be optionally replaced. １ ~C ６ Heteroalkyl, optionally substituted C ３ ~C １０ Cycloalkyl, optionally substituted 3-6 membered heterocyclyl, optionally substituted 5-10 membered heteroaryl, or optionally substituted C ６ ~C １０ It is Ariel, R １１ C is a hydrogen atom, optionally substituted. １ ~C ６ Alkyl, optionally substituted C １ ~C ６ Heteroalkyl, optionally substituted C ２ ~C ６ Alkenyl, C which can be optionally replaced. ２ ~C ６ Heteroalkenyl, C which is optionally substituted. ２ ~C ６ Alkinyl, C, which can be optionally replaced. ２ ~C ６ Heteroalkynyl, C which is optionally substituted. ３ ~C １０ Cycloalkyl, optionally substituted 3- to 10-membered heterocyclyl, optionally substituted C ３ ~C １０ Cycloalkenyls, optionally substituted 3- to 10-membered heterocycloalkenyls, optionally substituted C ６ ~C １０ [An aryl, or a 5- to 10-membered heteroaryl that is optionally substituted.]

18. The aforementioned compound is of formula VIIa-2: 【Chemistry 21】 A compound according to claim 17 having the structure, or a pharmaceutically acceptable salt thereof [in which case, R ６ C is a hydrogen atom, optionally substituted. １ ~C ６ Alkyl, optionally substituted C １ ~C ６ Heteroalkyls, optionally substituted 3-6 member heterocyclyls, optionally substituted 3-6 member cycloalkyls, optionally substituted 5-10 member heteroaryls, optionally substituted C ６ ~C １０ Ariel, 【Chemistry 22】 And, R ７ and R ８ Each of these can be independently replaced by H or C of any choice. １ ~C ６ It is alkyl, R ９ C is replaced by any choice. １ ~C ６ Alkyl, optionally substituted C ２ ~C ６ The alkenyl, optionally substituted 3-6 membered cycloalkyl, or optionally substituted 3-6 membered heterocyclyl, R １０ C is replaced by any choice. １ ~C ６ Alkyl, optionally substituted C ２ ~C ６ Alkenyl, C which can be optionally replaced. １ ~C ６ Heteroalkyl, optionally substituted C ３ ~C １０ Cycloalkyl, optionally substituted 3-6 membered heterocyclyl, optionally substituted 5-10 membered heteroaryl, or optionally substituted C ６ ~C １０ It is Ariel, R １１ C is a hydrogen atom, optionally substituted. １ ~C ６ Alkyl, optionally substituted C １ ~C ６ Heteroalkyl, optionally substituted C ２ ~C ６ Alkenyl, C which can be optionally replaced. ２ ~C ６ Heteroalkenyl, C which is optionally substituted. ２ ~C ６ Alkinyl, C, which can be optionally replaced. ２ ~C ６ Heteroalkynyl, C which is optionally substituted. ３ ~C １０ Cycloalkyl, optionally substituted 3- to 10-membered heterocyclyl, optionally substituted C ３ ~C １０ Cycloalkenyls, optionally substituted 3- to 10-membered heterocycloalkenyls, optionally substituted C ６ ~C １０ [An aryl, or a 5- to 10-membered heteroaryl that is optionally substituted.]

19. Compounds listed in Table 1 or Table 2, or pharmaceutically acceptable salts thereof.

20. A pharmaceutical composition comprising a compound according to any one of claims 1 to 19, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

21. A method for treating cancer in a subject requiring cancer treatment, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 19, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 20.

22. A method for treating a Ras protein-related disorder in a subject requiring treatment for the Ras protein-related disorder, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 19, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 20.

23. A method for inhibiting intracellular Ras protein, comprising contacting the cells with an effective amount of a compound according to any one of claims 1 to 19, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 20.

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