RAS inhibitors

Abstract

The invention relates to RAS inhibitors. The present invention features macrocyclic compounds capable of inhibiting the Ras protein as well as pharmaceutical compositions and protein complexes thereof, and their use in the treatment of cancer.

Description

[0001] This application is a divisional application of an invention patent application, the application date of the parent application is October 7, 2022, the application number is 202280080572.8 (PCT / US2022 / 077784), and the name of the invention is “RAS inhibitor”. Background Art

[0002] The vast majority of small molecule drugs work by binding to functionally important pockets on the target protein, thereby regulating the activity of the protein. For example, cholesterol-lowering drugs called statins bind to the enzyme active site of HMG-CoA reductase, thereby preventing the enzyme from engaging with its substrate. In fact, many such drug / target interaction pairs are known, which may mislead people to believe that small molecule regulators for most (if not all) proteins can be found to provide a reasonable amount of time, effort and resources. But this is far from the case. At present, it is estimated that only about 10% of all human proteins can be used as targets for small molecules. Bojadzic and Buchwald, Curr Top Med Chem 18: 674-699 (2019). The remaining 90% are currently considered to be difficult to treat or difficult to handle with the small molecule drug discovery mentioned above. Such targets are generally referred to as "undruggable". These undruggable targets include most of the medically important human proteins and usually undeveloped reservoirs. Therefore, people are very interested in discovering novel molecular modalities that can regulate the functions of such undruggable targets.

[0003] It is well established in the literature that Ras proteins (K-Ras, H-Ras, and N-Ras) play a critical role in a variety of human cancers and are therefore appropriate targets for anticancer therapy. In fact, in the United States, approximately 30% of all human cancers are caused by mutations in Ras proteins, many of which are lethal. Ras protein dysregulation due to activating mutations, overexpression, or upstream activation is common in human tumors, and activating mutations in Ras are frequently found in human cancers. For example, activating mutations at codon 12 in Ras proteins act by inhibiting GTPase activating protein (GAP) dependency and intrinsic GTP hydrolysis rates, significantly biasing the population of Ras mutant proteins toward the "on" (GTP-bound) state (Ras(ON)), leading to oncogenic MAPK signaling. Notably, Ras exhibits a picomolar affinity for GTP, allowing Ras to be activated even in the presence of low concentrations of this nucleotide. Mutations at codons 13 (eg, G13D) and 61 (eg, Q61K) in Ras also cause oncogenic activity in some cancers.

[0004] Despite extensive drug discovery efforts targeting Ras over the past decades, only one drug targeting the K-RasG12C mutant has been approved (sotorasib). More efforts are needed to discover other drugs targeting cancers driven by other Ras mutations. Summary of the invention

[0005] Ras inhibitors are provided herein. The methods described herein require the formation of a high-affinity three-component complex between a synthetic ligand and two intracellular proteins that do not interact under normal physiological conditions: a target protein of interest (e.g., Ras), and a cytoplasmic chaperone (presenting protein) (e.g., cyclophilin A) that is widely expressed in cells. More specifically, in some embodiments, the Ras inhibitors described herein induce new binding pockets in Ras by driving the formation of a high-affinity ternary complex or conjugate between the Ras protein and the widely expressed cytoplasmic chaperone cyclophilin A (CYPA). Without being bound by theory, the inventors believe that one way in which the compounds of the present invention and the complexes or conjugates formed therefrom exert an inhibitory effect on Ras is to spatially block the interaction sites between Ras and downstream effector molecules such as RAF and PI3K required to propagate oncogenic signals.

[0006] Thus, in some embodiments, the invention features a compound of formula I or a pharmaceutically acceptable salt thereof:

[0007]

[0008] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0009] X 1 , X 2 and X 3 Each independently selected from CH 2 CF 2 , C=O or O;

[0010] m is 1 or 2;

[0011] n is 0 or 1;

[0012] R 1 is hydrogen, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted 3- to 10-membered heterocycloalkyl;

[0013] R 2 is an optionally substituted C 1 -C 6Alkyl; and

[0014] R 3 is an optionally substituted C 1 -C 6 alkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted heterocycloalkyl,

[0015] and wherein each hydrogen is independently optionally isotopically enriched with deuterium.

[0016] In some embodiments, the invention features a compound having structural formula II or a pharmaceutically acceptable salt thereof:

[0017]

[0018] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0019] R 2 is an optionally substituted C 1 -C 6 Alkyl; and

[0020] R 3 is an optionally substituted C 1 -C 6 alkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted heterocycloalkyl,

[0021] and wherein each hydrogen is independently optionally isotopically enriched with deuterium.

[0022] In some embodiments, the invention features a compound having structural formula V or a pharmaceutically acceptable salt thereof:

[0023]

[0024] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0025] R 2 is an optionally substituted C 1 -C 6 Alkyl; and

[0026] R 3 is an optionally substituted C 1 -C 6 alkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted heterocycloalkyl,

[0027] and wherein each hydrogen is independently optionally isotopically enriched with deuterium.

[0028] In some embodiments, the invention features a compound having structural formula VI or a pharmaceutically acceptable salt thereof:

[0029]

[0030] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0031] R 2 is an optionally substituted C 1 -C 6 Alkyl; and

[0032] R 3 is an optionally substituted C 1 -C 6 alkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted heterocycloalkyl,

[0033] and wherein each hydrogen is independently optionally isotopically enriched with deuterium.

[0034] In some embodiments, the invention features a compound having structural formula VII or a pharmaceutically acceptable salt thereof:

[0035]

[0036] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0037] R 2 is an optionally substituted C 1 -C 6 Alkyl; and

[0038] R 3 is an optionally substituted C 1 -C 6 alkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted heterocycloalkyl,

[0039] and wherein each hydrogen is independently optionally isotopically enriched with deuterium.

[0040] In some embodiments, the invention also features a compound selected from Table 1 or Table 2, or a pharmaceutically acceptable salt thereof.

[0041] Also provided is a pharmaceutical composition comprising a compound of Formula I, Formula II, Formula V, Formula VI or Formula VII or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. Also provided is a pharmaceutical composition comprising a compound of Table 1 or Table 2 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0042] Also provided is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0043] In some embodiments, a method of treating a Ras protein-related disorder in a subject in need thereof is provided, the method comprising administering to the subject a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0044] Also provided is a method for inhibiting Ras protein in a cell, which comprises contacting the cell with an effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof.

[0045] Specifically, after careful consideration, any limitation discussed about one embodiment of the present invention may be applicable to any other embodiment of the present invention. In addition, any compound or composition of the present invention can be used in any method of the present invention, and any method of the present invention can be used to manufacture or utilize any compound or composition of the present invention.

[0046] Definitions and Chemical Terms

[0047] In this application, unless otherwise clear from the context, (i) the term "a" means "one or more"; (ii) the term "or" is used to mean "and / or" unless it is clearly indicated that the term refers to the alternative as the only one or the alternatives are mutually exclusive, however, the definition supported by the present invention refers to the only alternative and "and / or"; (iii) the terms "comprising" and "including" should be understood to cover the listed components or steps, whether present only the components or steps themselves or in combination with one or more additional components or steps; and (iv) when a range is provided, the endpoints are included.

[0048] As used herein, the term "about" is used to indicate that a value includes the standard deviation of the error of the device or method for determining the value. In certain embodiments, the term "about" refers to a range of values ​​in either direction (greater than or less than) 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or lower percentages, unless otherwise specified or otherwise apparent from the context (e.g., when the number will exceed 100% of the possible value).

[0049] As used herein, in the context of describing adjacent atoms, the term "adjacent" refers to divalent atoms that are directly connected by a covalent bond.

[0050] As used herein, "compounds of the present invention" and similar terms, whether explicitly indicated or not, refer to the Ras inhibitors described herein, including compounds of any of Formula I to Formula VII or subformulae thereof, as well as compounds of Table 1 or Table 2, and salts (e.g., pharmaceutically acceptable salts), solvates, hydrates, stereoisomers (including atropisomers) and tautomers thereof.

[0051] The term "wild type" refers to an entity having a structure or activity found in nature in a "normal" (as opposed to mutant, diseased, altered, etc.) state or situation. Those skilled in the art will appreciate that wild-type genes and polypeptides typically exist in a variety of different forms (e.g., alleles).

[0052] It will be appreciated by those skilled in the art 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., wherein one or more atoms are substituted with different isotopes of the atom, such as hydrogen being substituted with deuterium). Unless otherwise indicated or clear from the context, depicted structures may be understood to represent any such isomeric or isotopic forms, either individually or in combination.

[0053] The compounds described herein may be asymmetric (e.g., have one or more stereocenters). Unless otherwise indicated, all stereoisomers, such as enantiomers and diastereomers, are contemplated. Compounds of the present invention containing asymmetrically substituted carbon atoms may be separated in optically active form or racemic form. Methods for preparing optically active forms from optically active starting materials are known in the art, such as by resolving racemic mixtures or by stereoselective synthesis methods. Many geometric isomers such as olefins, C=N double bonds, etc. may also be present in the compounds described herein, and all such stable isomers are contemplated in the present invention. The cis and trans geometric isomers of the compounds of the present invention have been described and may be separated in the form of isomeric mixtures or in separate isomeric forms.

[0054] In some embodiments, one or more compounds described herein may exist in different tautomeric forms. It will be clear from the context that unless explicitly excluded, all such tautomeric forms are mentioned for such compounds. In some embodiments, the tautomeric form is obtained by exchanging a single bond with an adjacent double bond and accompanied by the migration of a proton. In certain embodiments, the tautomeric form may be a proton transfer tautomer, which is an isomer protonation state with the same empirical formula and total charge as the reference form. Examples of parts with proton transfer tautomeric forms include keto-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, amide-imidic acid pairs, enamine-imine pairs, and cyclic forms, in which protons may occupy two or more positions of a heterocyclic system, such as 1H-imidazole and 3H-imidazole, 1H-triazole, 2H-triazole and 4H-1,2,4-triazole, 1H-isoindole and 2H-isoindole, and 1H-pyrazole and 2H-pyrazole. In some embodiments, tautomeric forms may be in equilibrium or sterically locked into one form by appropriate substitution. In certain embodiments, tautomeric forms result from acetal interconversion.

[0055] Unless otherwise specified, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as 2 H. 3 H. 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 32 P. 33 P. 35 S. 18 F. 36 Cl, 123 I and 125 I. Isotope-labeled compounds (e.g. 3 H and 14 C) can be used in compound or substrate tissue distribution assays. Tritiated (i.e., 3 H) and carbon-14 (i.e., 14 C) isotopes are useful because they are easy to prepare and detect. Alternatively, heavier isotopes, such as deuterium (i.e., 2H) substitution may provide certain therapeutic benefits due to enhanced metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements). In some embodiments, one or more hydrogen atoms are replaced by 2 H or 3 H replacement, or one or more carbon atoms are 13 C or 14 C-enriched carbon replacement. Positron-emitting isotopes, such as 15 O. 13 N. 11 C and 18 F can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy. The preparation of isotopically labeled compounds is known to those skilled in the art. For example, isotopically labeled compounds can generally be prepared by following procedures similar to those disclosed for the compounds of the invention described herein, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0056] As used interchangeably herein, "deuterium substituted," "deuterated," or "deuterium enriched" refers to the deuterium (D or 2 In some embodiments, the minimum isotopic enrichment factor of the compositions of the invention is at least 5 (0.075% deuterium incorporation), such as at least 10 (0.15% deuterium incorporation). In other embodiments, the isotopic enrichment factor of the composition is at least 50 (0.75% deuterium incorporation), at least 500 (7.5% deuterium incorporation), at least 2000 (30% deuterium incorporation), at least 3000 (45% deuterium incorporation), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation) or at least 6600 (99% deuterium incorporation).

[0057] Non-limiting examples of moieties that may contain one or more deuterium substitutions in compounds of the invention include the following, wherein any position "R" may be deuterium (D):

[0058] Additional examples include the following:

[0059] And similar to R 1 Deuterated part of the type, where R 1The definition of W is found herein (e.g., in compounds of any of Formulae I-VII). Deuteration of moieties within a crosslinking group (e.g., an optionally substituted aziridine moiety) in compounds of the invention is also contemplated, wherein the crosslinking group is as defined herein (see, e.g., Formulae I-VII and subformulae thereof, and specific examples of W described herein, e.g.,

[0060] ). In addition, deuteration of available positions in any A moiety of the compounds of the formulae described herein is also contemplated, for example

[0061] Additionally, deuterium substitution may also occur at the linker position of the compounds of the invention and the various formulae described herein, for example

[0062]

[0063] In another embodiment, silylation substitutions are also contemplated, for example substitutions in the following linker positions:

[0064] Other examples of silylation include, for example, the following moieties:

[0065] And similar to R 1 Silylation substitution of the type part, where R 1 The definition of is found herein (eg, in a compound of any of Formulae I-VII).

[0066] As is known in the art, many chemical entities can present a variety of different solid forms, such as amorphous or crystalline forms (e.g., polymorphs, hydrates, solvates). In some embodiments, the compounds of the invention can be used in any such form, including any solid form. In some embodiments, the compounds described or depicted herein can be provided in the form of a hydrate or a solvate.

[0067] At various places in this specification, substituents of compounds of the invention are disclosed in groups or in ranges. Specifically, it is contemplated that the invention includes every individual subcombination of the members of the groups and ranges. For example, the term "C 1 -C 6 "Alkyl" specifically means to disclose individually methyl, ethyl, C 3 Alkyl, C 4 Alkyl, C 5 Alkyl and C 6In addition, where a compound includes multiple positions, and substituents at those positions are disclosed in groups or in ranges, unless otherwise indicated, the invention is intended to encompass individual compounds and groups of compounds (e.g., species and subclasses) containing every individual subcombination of members at each position.

[0068] The term "optionally substituted X" (e.g., "optionally substituted alkyl") is intended to be equivalent to "X, wherein X is optionally substituted" (e.g., "alkyl, wherein the alkyl is optionally substituted"). It is not intended to mean that the feature "X" (e.g., alkyl) itself is optional. As described herein, certain compounds 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 the designated moiety are replaced with a suitable substituent, such as any of the substituents or groups described herein. Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent at each position may be the same or different. For example, in the term "optionally substituted C 1 -C 6 Alkyl-C 2 -C 9 In the "heteroaryl" group, the alkyl portion, the heteroaryl portion, or both may be optionally substituted. Combinations of substituents envisioned by this invention are preferably those that form stable or chemically feasible compounds. As used herein, the term "stable" refers to a compound that is not substantially altered when subjected to conditions that allow the compound to be produced, detected, and in certain embodiments, recovered, purified, and used for one or more of the purposes disclosed herein.

[0069] Suitable monovalent substituents on the substitutable carbon atoms of the "optionally substituted" group may independently be deuterium; halogen; -(CH 2 ) 0-4 R o ;-(CH 2 ) 0-4 OR o ; -O(CH 2 ) 0-4 R o ;-O-(CH 2 ) 0 - 4 C(O)OR o ;-(CH 2 ) 0-4 CH(OR o ) 2 ;-(CH 2 ) 0-4 SR o ;-(CH 2 ) 0-4 Ph, this group can be R o Substitution; -(CH 2 ) 0-4 O(CH 2 ) 0-1 Ph, this group can be R o Substitution; -CH=CHPh, this group can be R o Substitution; -(CH 2 ) 0-4 O(CH 2 ) 0-1 -pyridyl, which can be o Substituted; 4- to 8-membered saturated or unsaturated heterocycloalkyl (eg, pyridyl); 3- to 8-membered saturated or unsaturated cycloalkyl (eg, cyclopropyl, cyclobutyl or cyclopentyl); -NO 2 ;-CN;-N 3 ;-(CH 2 ) 0 - 4 N(R o ) 2 ;-(CH 2 ) 0-4 N(R o )C(O)R o ;-N(R o )C(S)R o ;-(CH 2 ) 0 - 4 N(R o )C(O)NR o 2 ;-N(R o )C(S)NR o 2 ;-(CH 2 ) 0-4 N(R o )C(O)OR o ;-N(R o )N(R o )C(O)R o ;-N(R o )N(R o )C(O)NR o 2 ;-N(R o )N(R o )C(O)OR o ;-(CH 2 ) 0-4 C(O)R o;-C(S)R o ;-(CH 2 ) 0-4 C(O)OR o ;-(CH 2 ) 0-4 -C(O)-N(R o ) 2 ;-(CH 2 ) 0-4 -C(O)-N(R o )-S(O) 2 -R o ;-C(NCN)NR o 2 ;-(CH 2 ) 0-4 C(O)SR o ;-(CH 2 ) 0-4 C(O)OSiR o 3 ;-(CH 2 ) 0-4 OC(O)R o ;-OC(O)(CH 2 ) 0-4 SR o ;-SC(S)SR o ;-(CH 2 ) 0-4 SC(O)R o ;-(CH 2 ) 0-4 C(O)NR o 2 ;-C(S)NR o 2 ;-C(S)SR o ;-(CH 2 ) 0-4 OC(O)NR o 2 ;-C(O)N(OR o )R o ;-C(O)C(O)R o ;-C(O)CH 2 C(O)R o ;-C(NOR o )R o ;-(CH 2 ) 0-4 SSR o ;-(CH 2 ) 0-4 S(O) 2 R o ;-(CH2 ) 0-4 S(O) 2 OR o ;-(CH 2 ) 0-4 OS(O) 2 R o ; -S(O) 2 NR o 2 ;-(CH 2 ) 0-4 S(O)R o ;-N(R o )S(O) 2 NR o 2 ;-N(R o )S(O) 2 R o ;-N(OR o )R o ;-C(NOR o )NR o 2 ; -C(NH)NR o 2 ;-P(O) 2 R o ;-P(O)R o 2 ;-P(O)(OR o ) 2 ;-OP(O)R o 2 ;-OP(O)(OR o ) 2 ;-OP(O)(OR o )R v ;-SiR o 3 ;-(C 1 -C 4 Straight chain or branched chain alkylene)ON(R o ) 2 ; or -(C 1 -C 4 Straight or branched alkylene)C(O)ON(R o ) 2 , where each R o may be substituted as defined below and are independently hydrogen, -C 1 -C 6 Aliphatic group, -CH 2 Ph, -O(CH 2 ) 0-1 Ph, -CH 2-(5- to 6-membered heteroaryl ring), or a 3- to 6-membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or, notwithstanding the above definitions, two independently occurring R o Together with its intervening atoms it forms a 3 to 12 membered saturated, partially unsaturated or aromatic mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, which may be substituted as defined below.

[0070] R o (or two independent R o Suitable monovalent substituents on the ring formed with its intervening atoms) may independently be halogen, -(CH 2 ) 0-2 R ● 、-(halogenated R ● )、-(CH 2 ) 0-2 OH, -(CH 2 ) 0-2 OR ● 、-(CH 2 ) 0-2 CH(OR ● ) 2 、-O(halogenated R ● ), -CN, -N 3 、-(CH 2 ) 0 - 2 C(O)R ● 、-(CH 2 ) 0-2 C(O)OH, -(CH 2 ) 0-2 C(O)OR ● 、-(CH 2 ) 0-2 SR ● 、-(CH 2 ) 0 - 2 SH, -(CH 2 ) 0-2 NH 2 、-(CH 2 ) 0-2 NHR ● 、-(CH 2 ) 0-2 NR ● 2 、-NO 2 、-SiR ● 3 、-OSiR ● 3 、-C(O)SR ● 、-(C1-4 Straight or branched chain alkylene)C(O)OR ● or -SSR ● , where each R ● unsubstituted or, when preceded by "halo", substituted only by one or more halogens, and independently selected from C 1 -C 4 Aliphatic group, -CH 2 Ph, -O(CH 2 ) 0-1 Ph or a 5- to 6-membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. o Suitable divalent substituents on a saturated carbon atom of include =0 and =S.

[0071] Suitable divalent substituents on a saturated carbon atom of an "optionally substituted" group include the following: =0, =S, =NNR * 2 、=NNHC(O)R * 、=NNHC(O)OR * 、=NNHS(O) 2 R * , =NR * , =NOR * 、-O(C(R * 2 )) 2-3 O-or-S(C(R * 2 )) 2-3 S-, where R * is selected at each independent occurrence from hydrogen; C 1 -C 6 an aliphatic group which may be substituted as defined below; or an unsubstituted 5- to 6-membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Suitable divalent substituents attached to an adjacent substitutable carbon of an "optionally substituted" group include: -O(CR * 2 ) 2- 3 O-, where R * is selected at each independent occurrence from hydrogen; C 1 -C 6 an aliphatic group which may be substituted as defined below; or an unsubstituted 5- to 6-membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0072] R * Suitable substituents on the aliphatic group include halogen, -R ● 、-(halogenated R● ), -OH, -OR ● 、-O(halogenated R ● )、-CN、-C(O)OH、-C(O)OR ● 、-NH 2 、-NHR ● 、-NR ● 2 or-NO 2 , where each R ● is unsubstituted or, when preceded by "halo", is substituted only by one or more halogens, and is independently C 1 -C 4 Aliphatic group, -CH 2 Ph, -O(CH 2 ) 0 - 1 Ph or a 5- to 6-membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0073] Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include or Among them are independently hydrogen; C 1 -C 6 an aliphatic group which may be substituted as defined below; unsubstituted -OPh; or an unsubstituted 3- to 6-membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or, notwithstanding the above definitions, two independently present Together with its intervening atoms, it forms a 3- to 12-membered saturated, partially unsaturated or aromatic monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0074] Suitable substituents on the aliphatic group are independently halogen, -R ● 、-(halogenated R ● ), -OH, -OR ● 、-O(halogenated R ● )、-CN、-C(O)OH、-C(O)OR ● 、-NH 2 、-NHR ● 、-NR ● 2 or-NO 2 , where each R ● is unsubstituted or, when preceded by "halo", is substituted only by one or more halogens, and is independently C 1 -C4 Aliphatic group, -CH 2 Ph, -O(CH 2 ) 0-1 Ph or a 5- to 6-membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Suitable divalent substituents on a saturated carbon atom of include =0 and =S.

[0075] As used herein, the term "acetyl" refers to the group -C(O)CH 3 .

[0076] As used herein, the term "alkoxy" refers to -OC 1 -C 20 Alkyl groups, wherein the alkoxy group is attached to the rest of the compound through an oxygen atom.

[0077] As used herein, the term "alkyl" refers to a saturated, straight or branched monovalent hydrocarbon radical containing 1 to 20 (e.g., 1 to 10 or 1 to 6) carbons. In some embodiments, the alkyl group is unbranched (i.e., linear); in some embodiments, the alkyl group is branched. Alkyl groups are, for example, but not limited to, methyl, ethyl, n-propyl and isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl, and neopentyl.

[0078] As used herein, the term "alkylene" means a saturated divalent hydrocarbon group derived from a straight or branched chain saturated hydrocarbon by removing two hydrogen atoms, and examples thereof are methylene, ethylene, isopropylene, and the like. x -C y "Alkylene" means an alkylene group having between x and y carbons. Exemplary x values ​​are 1, 2, 3, 4, 5, and 6, and exemplary y values ​​are 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, or 20 (e.g., C 1 -C 6 , C 1 -C 10 , C 2 -C 20 , C 2 -C 6 , C 2 -C 10 or C 2 -C 20 In some embodiments, the alkylene group may be further substituted with 1, 2, 3, or 4 substituents as defined herein.

[0079] Unless otherwise specified, as used herein, the term "alkenyl" means a monovalent straight or branched chain group having 2 to 20 carbons (e.g., 2 to 6 or 2 to 10 carbons) containing one or more carbon-carbon double bonds, and examples thereof are vinyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, and 2-butenyl. Alkenyl includes both cis and trans isomers. Unless otherwise specified, as used herein, the term "alkenylene" means a divalent straight or branched chain group having 2 to 20 carbons (e.g., 2 to 6 or 2 to 10 carbons) containing one or more carbon-carbon double bonds.

[0080] As used herein, the term "alkynyl" refers to a monovalent straight or branched chain group having 2 to 20 carbons (eg, 2 to 4, 2 to 6, or 2 to 10 carbons) containing a carbon-carbon triple bond, and examples thereof are ethynyl and 1-propynyl.

[0081] As used herein, the term "amino" refers to For example -NH 2 and -N(CH 3 ) 2 .

[0082] 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.

[0083] As used herein, the term "amino acid" refers to a amino acid having a side chain, an amino group and an acid group (e.g., -CO 2 H or -SO 3 H) wherein the amino acid is attached to the parent molecular group via the side chain, amino group or acid group (e.g., side chain). As used herein, the term "amino acid" refers in the broadest sense to any compound or substance that can be incorporated into a polypeptide chain, for example, by forming one or more peptide bonds. In some embodiments, an amino acid has a general structure H 2 NC(H)(R)-COOH. In some embodiments, the amino acid is a naturally occurring amino acid. In some embodiments, the amino acid is a synthetic amino acid; in some embodiments, the amino acid is a D-amino acid; in some embodiments, the amino acid is an L-amino acid. "Standard amino acid" refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides. Exemplary amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, optionally substituted hydroxynorvaline, isoleucine, leucine, lysine, methionine, norvaline, ornithine, phenylalanine, proline, pyrrolysine, selenocysteine, serine, taurine, threonine, tryptophan, tyrosine, and valine.

[0084] As used herein, the term "aryl" refers to a monovalent monocyclic, bicyclic or polycyclic system formed by carbon atoms, wherein the ring connected to the side-joining group is an aromatic ring. Examples of aryl groups include phenyl, naphthyl, phenanthrenyl and anthracenyl. The aromatic ring can be connected to its side-joining group at any heteroatom or carbocyclic atom that produces a stable structure, and unless otherwise specifically stated, any ring atom may be optionally substituted.

[0085] As used herein, the term “C 0 " represents a bond. For example, the term -N(C(O)-(C 0 -C 5 The alkylene-H)- moiety includes -N(C(O)-(C 0 Alkylene-H)-, also represented by -N(C(O)-H)-.

[0086] As used herein, the terms "carbocycle" and "carbocyclyl" refer to monovalent, optionally substituted 3 to 12-membered monocyclic, bicyclic or tricyclic structures, which may be bridged, fused or spirocyclic rings, wherein all rings are formed by carbon atoms, and at least one ring is a non-aromatic ring. Carbocyclic structures include cycloalkyl, cycloalkenyl and cycloalkynyl. Examples of carbocyclic groups include cyclohexyl, cyclohexenyl, cyclooctynyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, fluorenyl, indenyl, dihydroindenyl, decahydronaphthyl, etc. Carbocycles may be connected to their side groups at any ring atom producing a stable structure, and unless otherwise specifically stated, any ring atom may be optionally substituted.

[0087] As used herein, the term "carbonyl" refers to a C(O) group, which may also be represented as C=O.

[0088] As used herein, the term "carboxyl" means -CO 2 H, (C=O)(OH), COOH or C(O)OH, or the unprotonated counterpart.

[0089] As used herein, the term "cyano" refers to a -CN group.

[0090] As used herein, the term "cycloalkyl" refers to a monovalent saturated cyclic hydrocarbon group, which, unless otherwise specifically stated, may be a bridged ring, a fused ring or a spiro ring having three to eight ring carbons, and examples thereof are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cycloheptyl.

[0091] As used herein, the term "cycloalkenyl" refers to a monovalent, non-aromatic, saturated cyclic hydrocarbon group which, unless otherwise specifically stated, may be a bridged, fused, or spiro ring having three to eight ring carbons and containing one or more carbon-carbon double bonds.

[0092] As used herein, the term "diastereoisomers" means stereoisomers that are not mirror images of each other and are not superimposable on each other.

[0093] As used herein, the term "enantiomer" means each individual optically active form of a compound of the invention having an optical purity or enantiomeric excess (as determined by standard methods in the art) of at least 80% (i.e., at least 90% of one enantiomer and up to 10% of the other enantiomer), preferably at least 90% and more preferably at least 98%.

[0094] As used herein, the term "haloacetyl" refers to an acetyl group in which at least one hydrogen is replaced by a halogen.

[0095] As used herein, the term "haloalkyl" refers to an alkyl moiety in which one or more carbon atoms are substituted with one or more same or different halogen moieties.

[0096] As used herein, the term "halogen" refers to a halogen selected from bromine, chlorine, iodine or fluorine.

[0097] As used herein, the term "heteroalkyl" refers to an "alkyl" as defined herein in which at least one carbon atom is replaced by a heteroatom (eg, O, N or S atom). The heteroatom may be present in the middle or at the terminal of the group.

[0098] As used herein, the term "heteroaryl" refers to a monovalent, monocyclic or polycyclic structure containing at least one completely aromatic ring: that is, it contains 4n+2 π electrons in the monocyclic or polycyclic system and contains at least one heteroatom selected from N, O or S in the aromatic ring. Exemplary unsubstituted heteroaryl 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) carbons. The term "heteroaryl" includes any of the above heteroaromatic rings fused to one or more aromatic or carbon rings, such as a phenyl ring or a cyclohexane ring. Examples of heteroaryl include, but are not limited to, pyridyl, pyrazolyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, imidazolyl, thiazolyl, quinolyl, tetrahydroquinolyl and 4-azaindolyl. The heteroaryl ring may be attached to its pendant group at any ring atom that results in a stable structure, and unless otherwise specifically stated, any ring atom may be optionally substituted. In a certain embodiment, the heteroaryl group is substituted with 1, 2, 3 or 4 substituents.

[0099] As used herein, the term "heterocycloalkyl" means a monovalent, monocyclic, bicyclic or polycyclic system in which at least one ring is non-aromatic and the non-aromatic ring contains one, two, three or four heteroatoms independently selected from the group consisting of nitrogen, oxygen and sulfur, which can be a bridged ring, a fused ring or a spiro ring. The 5-membered ring has zero to two double bonds, and the 6-membered ring and the 7-membered ring have zero to three double bonds. Exemplary unsubstituted heterocycloalkyls 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) carbons. The term "heterocycloalkyl" also means a heterocyclic compound with a bridged polycyclic structure, in which one or more carbons or heteroatoms bridge non-adjacent members of a monocyclic ring, such as a quinuclidine radical. The term "heterocycloalkyl" includes bicyclic, tricyclic and tetracyclic groups of any of the above heterocycles fused with one or more aromatic, carbocyclic, heteroaromatic or heterocyclic rings, such as aryl, cyclohexane, cyclohexene, cyclopentane, cyclopentene, pyridine or pyrrolidine rings. Examples of heterocycloalkyls include pyrrolidinyl, piperidinyl, 1,2,3,4-tetrahydroquinolinyl, decahydroquinolinyl, dihydropyrrolopyridinyl and decahydronaphthyridinyl. The heterocycloalkyl ring may be attached to its side group at any ring atom that results in a stable structure, and unless otherwise specifically stated, any ring atom may be optionally substituted.

[0100] As used herein, the term "hydroxy" refers to an -OH group.

[0101] 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.

[0102] As used herein, the term "isomer" means any tautomer, stereoisomer, atropisomer, enantiomer or diastereomer of any compound of the present invention. It should be recognized that the compounds of the present invention may have one or more chiral centers or double bonds and, therefore, exist in stereoisomeric form, such as double bond isomers (i.e., E / Z geometric isomers) or diastereomers (e.g., enantiomers (i.e., (+) or (-)) or cis / trans isomers). According to the present invention, the chemical structures depicted herein and therefore the compounds of the present invention encompass all corresponding stereoisomers, i.e., stereoisomerically pure (e.g., geometrically pure, enantiomerically pure or diastereomerically pure) form as well as enantiomers and stereoisomer mixtures, such as racemates. Enantiomeric and stereoisomeric mixtures of the compounds of the invention can be typically separated into their component enantiomers or stereoisomers by well-known methods, such as chiral phase gas chromatography, chiral phase high performance liquid chromatography, crystallization of the compound in the form of a chiral salt complex, or crystallization of the compound in a chiral solvent. Enantiomers and stereoisomers can also be obtained from stereoisomerically pure or enantiomerically pure intermediates, reagents and catalysts by well-known asymmetric synthesis methods.

[0103] As used herein, a "monovalent organic moiety" is less than 500 kDa. In some embodiments, a "monovalent organic moiety" is less than 400 kDa. In some embodiments, a "monovalent organic moiety" is less than 300 kDa. In some embodiments, a "monovalent organic moiety" is less than 200 kDa. In some embodiments, a "monovalent organic moiety" is less than 100 kDa. In some embodiments, a "monovalent organic moiety" is less than 50 kDa. In some embodiments, a "monovalent organic moiety" is less than 25 kDa. In some embodiments, a "monovalent organic moiety" is less than 20 kDa. In some embodiments, a "monovalent organic moiety" is less than 15 kDa. In some embodiments, a "monovalent organic moiety" is less than 10 kDa. In some embodiments, a "monovalent organic moiety" is less than 1 kDa. In some embodiments, a "monovalent organic moiety" is less than 500 g / mol. In some embodiments, a "monovalent organic moiety" is in the range between 500 g / mol and 500 kDa.

[0104] As used herein, the term "stereoisomer" refers to all possible different isomeric and conformational forms that a compound (e.g., a compound of any formula described herein) may have, particularly all possible stereochemical and conformational isomeric forms of the basic molecular structure, all diastereomers, enantiomers or conformational isomers, including atropisomers. Some compounds of the present invention may exist in different tautomeric forms, all of which are included within the scope of the present invention.

[0105] As used herein, the term "sulfonyl" refers to -S(O) 2 -group.

[0106] As used herein, the term "thiocarbonyl" refers to a -C(S)- group.

[0107] Those skilled in the art reading this disclosure will appreciate that certain compounds described herein may be provided or utilized in any of a variety of forms, such as salt forms, protected forms, prodrug forms, ester forms, isomeric forms (e.g., optical or structural isomers), isotopic forms, and the like. In some embodiments, a reference to a particular compound may refer to a particular form of that compound. In some embodiments, a reference to a particular compound may refer to that 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 that compound rather than a racemic mixture of that compound; a particular salt of a compound may be considered a different form from another salt of that compound; a preparation containing one configurational isomer of a double bond ((Z) or (E)) may be considered a different form from a preparation containing another configurational isomer of that double bond ((E) or (Z)); a preparation having one or more isotopes of an atom different from the isotopes present in a reference preparation may be considered to be in a different form. BRIEF DESCRIPTION OF THE DRAWINGS

[0109] Figure 1 The results show that the covalent KRAS G12D inhibitors Compound A and Compound B, which are representative of the compounds of the present invention, can inhibit the growth of HPACKRAS in human pancreatic adenocarcinoma. G12D / wt Strong and persistent RAS pathway modulation was demonstrated in a mouse xenograft model. RAS / ERK signaling pathway modulation was assessed by measuring human DUSP6 mRNA levels in a qPCR assay. Both Compound A and Compound B caused suppression of DUSP6 mRNA levels in HPAC xenograft tumors 4 hours after administration, indicating strong RAS pathway modulation.

[0110] Figure 2 Compound A and Compound B showed strong tumor cross-linking at 4 hours and up to 24 hours, consistent with significant DUSP6 inhibition. Figure 1 Tumor samples collected in the assay were homogenized to extract proteins. Protein lysates were subjected to Western blotting using Ras rabbit mAb (Abcam ab108602) and β-actin mAb (CST-4967). The appearance of a higher molecular weight (MW) band (cross-linked KRAS G12D band) in the tumor samples was detected, in which case the compound was covalently bound to the KRAS G12D protein.

[0111] Figure 3A We showed that in the HPAC CDX mouse xenograft model with heterozygous KRAS G12D, single-agent Compound A and Compound B administered orally at 100 mg / kg daily caused regression of all tumors at the end of treatment (day 28 after initiation of treatment) (regression defined as >10% tumor regression relative to baseline).

[0112] Figure 3B The results showed that no weight loss was observed with compound A (100 mg / kg po qd) or compound B (100 mg / kg po qd), indicating that the KRAS G12D / wt Both compounds were well tolerated at 100 mg / kg in the mouse xenograft model.

[0113] Figure 3C HPACKRAS in human pancreatic adenocarcinoma G12D / wt In the mouse xenograft model, 8 / 10 tumors and 9 / 10 tumors in the compound A group (100 mg / kg po qd) and compound B group (100 mg / kg po qd), respectively, achieved complete regression on day 28 (complete regression was defined as >85% tumor regression relative to baseline). DETAILED DESCRIPTION

[0114] Compound

[0115] Ras inhibitors are provided herein. The methods described herein require the formation of a high-affinity three-component complex or conjugate between a synthetic ligand and two intracellular proteins that do not interact under normal physiological conditions: a target protein of interest (e.g., Ras), and a cytoplasmic chaperone (presenting protein) (e.g., cyclophilin A) that is widely expressed in cells. More specifically, in some embodiments, the Ras inhibitors described herein induce new binding pockets in Ras by driving the formation of a high-affinity ternary complex or conjugate between the Ras protein and the widely expressed cytoplasmic chaperone cyclophilin A (CYPA). Without being bound by theory, the inventors believe that one way in which the compounds of the present invention and the complexes or conjugates formed therefrom exert an inhibitory effect on Ras is to spatially block the interaction sites between Ras and downstream effector molecules such as RAF required to propagate oncogenic signals.

[0116] Without being bound by theory, the inventors hypothesize that covalent and non-covalent interactions of the compounds of the invention with Ras and chaperone proteins (e.g., cyclophilin A) may contribute to the inhibition of Ras activity. In some embodiments, the compounds of the invention interact with the side chains of Ras proteins (e.g., -CH 2 -COOH or -CH 2-COO- side chain) to form covalent adducts. Covalent adducts can also be formed with other side chains of Ras. Additionally or alternatively, non-covalent interactions may work: for example, van der Waals interactions, hydrophobic interactions, hydrophilic interactions, and hydrogen bonding interactions, and combinations thereof can contribute to the ability of the compounds of the present invention to form complexes and act as Ras inhibitors. Therefore, the compounds of the present invention can inhibit a variety of Ras proteins (e.g., K-Ras, N-Ras, H-Ras, and mutants thereof mutated at positions 12, 13, and 61, such as G12C, G12D, G12V, G12S, G13C, G13D, and Q61L, as well as other mutants described herein).

[0117] Methods for determining covalent adduct formation are known in the art. One method for determining covalent adduct formation is to perform a "cross-linking" assay, such as the assay described below:

[0118] NOTE: The following protocol describes a procedure for monitoring the cross-linking of K-Ras G12D (GMP-PNP) with compounds of the invention. This protocol can also be adapted to other Ras proteins or nucleotides.

[0119] The purpose of this biochemical assay is to measure the ability of test compounds to covalently label nucleotide-loaded K-Ras isoforms. 2 A 5 μM stock solution of K-Ras(1-169)G12D loaded with GMP-PNP was diluted 10-fold to a final concentration of 0.5 μM in assay buffer containing 5 μM cyclophilin A and 2 μM test compound; and the final sample volume was 100 μL.

[0120] At 25 ° C, the samples were incubated for a period of up to 24 hours and then quenched by adding 10 μL of 5% formic acid. The quenched samples were centrifuged at 15000 rpm for 15 minutes in a benchtop centrifuge, and then 10 μL aliquots were injected onto a reverse phase C4 column and eluted into a mass spectrometer with an increasing acetonitrile gradient in the mobile phase. Analysis of the raw data can be performed using Waters MassLynxMS software, where the binding % is calculated from the deconvoluted protein peaks of labeled and unlabeled K-Ras.

[0121] Therefore, the present invention provides a compound having a structure of Formula 0 or a pharmaceutically acceptable salt thereof:

[0122]

[0123] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0124] X 1 , X 2 and X 3 Each independently selected from CH 2 CF 2 , C=O or O;

[0125] m is 1 or 2;

[0126] n is 0 or 1;

[0127] R 1 is hydrogen, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted 3- to 10-membered heterocycloalkyl;

[0128] R 2 is an optionally substituted C 1 -C 6 Alkyl; and

[0129] R 3 is an optionally substituted C 1 -C 6 alkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted heterocycloalkyl,

[0130] and wherein each hydrogen is independently optionally isotopically enriched with deuterium.

[0131] In some embodiments, the compounds of the present invention have the structure of Formula I, or a pharmaceutically acceptable salt thereof:

[0132]

[0133] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0134] X 1 , X 2 and X 3 Each independently selected from CH 2 CF 2 , C=O or O;

[0135] m is 1 or 2;

[0136] n is 0 or 1;

[0137] R 1 is hydrogen, optionally substituted C 1-C 6 heteroalkyl or optionally substituted 3- to 10-membered heterocycloalkyl;

[0138] R 2 is an optionally substituted C 1 -C 6 Alkyl; and

[0139] R 3 is an optionally substituted C 1 -C 6 alkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted heterocycloalkyl,

[0140] and wherein each hydrogen is independently optionally isotopically enriched with deuterium.

[0141] In some embodiments, the compounds of the present invention have the structure of Formula Ia, Formula Ib, Formula Ic, or a pharmaceutically acceptable salt thereof:

[0142]

[0143]

[0144] wherein each D indicates hydrogen having an isotopic enrichment factor of at least 5 with deuterium.

[0145] In some embodiments, the compounds of the present invention have the structure of Formula II, or a pharmaceutically acceptable salt thereof:

[0146]

[0147] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0148] R 2 is an optionally substituted C 1 -C 6 Alkyl; and

[0149] R 3 is an optionally substituted C 1 -C 6 alkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted heterocycloalkyl,

[0150] and wherein each hydrogen is independently optionally isotopically enriched with deuterium.

[0151] In some embodiments, the compounds of the invention have the structure of Formula V, or a pharmaceutically acceptable salt thereof:

[0152]

[0153] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0154] R 2 is an optionally substituted C 1 -C 6 Alkyl; and

[0155] R 3 is an optionally substituted C 1 -C 6 alkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted heterocycloalkyl,

[0156] and wherein each hydrogen is independently optionally isotopically enriched with deuterium.

[0157] In some embodiments, the compounds of the invention have the structure of Formula VI, or a pharmaceutically acceptable salt thereof:

[0158]

[0159] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0160] R 2 is an optionally substituted C 1 -C 6 Alkyl; and

[0161] R 3 is an optionally substituted C 1 -C 6 alkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted heterocycloalkyl,

[0162] and wherein each hydrogen is independently optionally isotopically enriched with deuterium.

[0163] In some embodiments, the compounds of the invention have the structure of Formula VII, or a pharmaceutically acceptable salt thereof:

[0164]

[0165] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0166] R 2 is an optionally substituted C 1 -C 6 Alkyl; and

[0167] R 3 is an optionally substituted C 1 -C 6 alkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted heterocycloalkyl,

[0168] and wherein each hydrogen is independently optionally isotopically enriched with deuterium.

[0169] In some embodiments, the compounds of the present invention have the structure of Formula Va, Formula Vb, Formula Vc, or a pharmaceutically acceptable salt thereof:

[0170]

[0171] wherein each D indicates hydrogen having an isotopic enrichment factor of at least 5 with deuterium.

[0172] In some embodiments, the compounds of the invention have the structure of Formula Vd, Formula Ve, Formula Vf, or a pharmaceutically acceptable salt thereof:

[0173]

[0174] wherein each D indicates hydrogen having an isotopic enrichment factor of at least 5 with deuterium.

[0175] In some embodiments, A is an optionally substituted thiazole-diyl, an optionally substituted oxazole-diyl, an optionally substituted morpholine-diyl, an optionally substituted pyrrolidine-diyl, an optionally substituted piperidine-diyl, or an optionally substituted phenylene. In some embodiments, A is an optionally substituted thiazole-diyl or an optionally substituted morpholine-diyl. In some embodiments of the compounds of the invention, A is an optionally substituted 5 to 10 membered heteroaryl. In some embodiments, A is: In some embodiments, A is

[0176] In some embodiments of the compounds of the invention, A is an optionally substituted phenylene. In some embodiments, A is: In some embodiments, A is

[0177] In some embodiments of the compounds of the present invention, A is an optionally substituted 3 to 6 membered heterocycloalkylene. In some embodiments, A is an optionally substituted 6 membered heterocycloalkylene. In some embodiments, A is selected from the following or a stereoisomer thereof: In some embodiments, A is selected from the following or stereoisomers thereof:

[0178] In some embodiments of the compounds of the present invention, R 1 is hydrogen or optionally substituted 3 to 10 membered heterocycloalkyl. 1 is an optionally substituted 3- to 10-membered heterocycloalkyl. 1 yes:

[0179] In some embodiments of the compounds of the present invention, R 1 yes: wherein each D indicates hydrogen having an isotopic enrichment factor of at least 5 with deuterium.

[0180] In some embodiments of the compounds of the present invention, R 2 yes:

[0181] In some embodiments of the compounds of the present invention, R 2 yes: and wherein each D indicates hydrogen having an isotopic enrichment factor of at least 5 with deuterium.

[0182] In some embodiments of the compounds of the present invention, R 3 is an optionally substituted C 1 -C 6 In some embodiments of the compounds of the present invention, R 3 is an optionally substituted C 1 -C 6 In some embodiments, R 3 yes: In some embodiments, R 3 yes: In some embodiments, R 3 yes: and wherein each D indicates hydrogen having an isotopic enrichment factor of at least 5 with deuterium.

[0183] In some embodiments of the compounds of the present invention, R 3 is or is optionally substituted 3 to 6 membered cycloalkyl. 3 yes: In some embodiments, R 3 yes:

[0184] In some embodiments of the compounds of the present invention, R 2 yes R 3 yes And A is

[0185] In some embodiments, R 2 yes R 3 yes And A is

[0186] In some embodiments of the compounds of the invention, m is 1. In some embodiments, n is 1. In some embodiments, X 1 Yes CH 2 In some embodiments, X 2 Yes CH 2 In some embodiments, X 3 Yes CH 2 In some embodiments, m is 1, n is 1, and X 1 , X 2 and X 3 Each is CH 2 .

[0187] In some embodiments, the compound of the present invention is selected from Table 1 or a pharmaceutically acceptable salt or stereoisomer thereof. In some embodiments, the compound of the present invention is selected from Table 1 or a pharmaceutically acceptable salt or atropisomer thereof.

[0188] Table 1: Certain compounds of the present invention

[0189]

[0190]

[0191]

[0192]

[0193]

[0194]

[0195]

[0196]

[0197]

[0198]

[0199]

[0200]

[0201]

[0202]

[0203] It should be noted that the bonds in some compounds are shown as straight lines or wedges. In some cases, the relative stereochemistry of stereoisomers has been determined; in some cases, the absolute stereochemistry has been determined. In some cases, a single example number corresponds to a mixture of stereoisomers. All stereoisomers of the above table compounds are contemplated in the present invention. In certain embodiments, atropisomers of the above table compounds are contemplated.

[0204] In some embodiments, a compound of Table 2 or a pharmaceutically acceptable salt thereof is provided. In some embodiments, the compound of the invention is selected from Table 2 or a pharmaceutically acceptable salt or atropisomer thereof.

[0205] Table 2: Certain compounds of the present invention

[0206]

[0207]

[0208]

[0209]

[0210]

[0211]

[0212]

[0213] It should be noted that the bonds in some compounds are shown as straight lines or wedges. In some cases, a single example number corresponds to a stereoisomer mixture. All stereoisomers of the above table compounds are encompassed in the present invention. In certain embodiments, atropisomers of the above table compounds are encompassed.

[0214] In some embodiments, the compound of the present invention is a compound selected from Table 3 or a pharmaceutically acceptable salt or stereoisomer thereof. In some embodiments, the compound of the present invention is a compound selected from Table 3 or a pharmaceutically acceptable salt or atropisomer thereof.

[0215] In some embodiments, the compound of the invention is not a compound selected from Table 3. The compound of the invention is not a compound selected from Table 3 or a pharmaceutically acceptable salt or stereoisomer thereof. In some embodiments, the compound of the invention is not a compound selected from Table 3 or a pharmaceutically acceptable salt or atropisomer thereof.

[0216] Table 3. Some compounds

[0217]

[0218]

[0219]

[0220] In some embodiments, the compounds of the invention include a cross-linking group (e.g., an optionally substituted aziridine moiety) that is bound to an organic moiety that is a Ras binding moiety, wherein after the compound is contacted with the Ras protein, the compound binds to the Ras protein to form a conjugate. For example, the cross-linking group (e.g., an optionally substituted aziridine moiety) of the compound can be bound to an amino acid of the Ras protein, such as cross-linked, to form a conjugate. In some embodiments, the Ras binding moiety is a K-Ras binding moiety. In some embodiments, the K-Ras binding moiety binds to the residues of the K-Ras Switch-II binding pocket of the K-Ras protein. In some embodiments, the Ras binding moiety is an H-Ras binding moiety that binds to the residues of the H-Ras Switch-II binding pocket of the H-Ras protein. In some embodiments, the Ras binding moiety is an N-Ras binding moiety that binds to the residues of the N-Ras Switch-II binding pocket of the N-Ras protein. The molecular weight of the Ras binding moiety is typically less than 1200Da. For a description of the various domains of the Ras protein, see, for example, Johnson et al., 292: 12981-12993 (2017), which is incorporated herein by reference.

[0221] In some embodiments, the compounds of the invention are or are used as prodrugs, such as for administration to a cell or a subject in need thereof.

[0222] Also provided is a pharmaceutical composition comprising a compound of the invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0223] Also provided is a conjugate or a salt thereof, wherein the conjugate or the salt thereof comprises a structure of formula III:

[0224] MP 1

[0225] Formula III

[0226] Where P 1 is a monovalent organic fraction; and

[0227] M has the structure of Formula IV:

[0228]

[0229] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0230] X 1 , X 2 and X 3 Each independently selected from CH 2 CF 2 , C=O or O;

[0231] m is 1 or 2;

[0232] n is 0 or 1;

[0233] R 1 is hydrogen, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted 3- to 10-membered heterocycloalkyl;

[0234] R 2 is an optionally substituted C 1 -C 6 Alkyl; and

[0235] R 3 is an optionally substituted C 1 -C 6 alkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted heterocycloalkyl,

[0236] and wherein each hydrogen of Formula IV is independently optionally isotopically enriched with deuterium.

[0237] In some embodiments of the conjugates of the invention, the monovalent organic moiety is a protein. In some embodiments, the protein is a Ras protein. In some embodiments, the Ras protein is K-Ras G12D or K-Ras G13D. In some embodiments of the conjugates of the invention, M is bound to an amino acid residue of the monovalent organic moiety.

[0238] In some embodiments, the compounds of the invention have improved oral bioavailability (%F) compared to compounds known in the art. Methods for measuring oral bioavailability are known in the art, and one such method is provided below:

[0239] Oral bioavailability can be determined in BALB / c mice. After intravenous (IV) bolus and oral gavage (PO) administration of test compounds, approximately 30 μL whole blood samples were collected at designated time points into a K-containing 2EDTA tubes. For some compounds, blood samples were centrifuged at 4600 rpm for about 5 minutes at 4°C and plasma samples were stored at -80°C prior to bioanalysis. Blood or plasma samples were extracted by protein precipitation and analyzed by tandem mass spectrometry (UPLC MS / MS) using positive ion electrospray ionization on, for example, an AB Sciex Triple Quad 6500+ mass spectrometer coupled to an Acquity UPLC system.

[0240] All PK parameters were obtained from blood (or plasma) concentration versus time data using non-compartmental analysis using WinNonlin. Bioavailability (F%, also written as %F) was estimated using the following formula:

[0241]

[0242] AUC inf,PO It is the area under the blood (or plasma) concentration curve from time zero to infinity following PO administration.

[0243] AUC inf,IV It is the area under the blood (or plasma) concentration curve from time zero to infinity following IV administration.

[0244] dose IV is the total dose administered IV

[0245] dose PO is the total dose administered PO

[0246] Generally speaking, F% (or %F) values ​​exceeding 10% are preferred.

[0247] Of the 19 compounds in Table 1 described herein that were tested for oral bioavailability, all but three compounds had a %F greater than 10%. Additionally, all but three compounds cross-linked greater than 60% of the K-Ras G12D residues over a 6 hour period in the biochemical cross-linking assay described herein. Thirteen of the compounds tested had a %F greater than 10% and cross-linked greater than 60% of the K-Ras G12D residues. Without being bound by theory, the inventors propose that the N-methylaziridine moiety of the compounds described herein is primarily responsible for this unexpected activity.

[0248] In some embodiments, the compounds of the invention are selective for one or more specific Ras mutants relative to other Ras mutants or wild type compared to compounds known in the art. Methods for measuring this selectivity are known in the art, such as Ras-Raf binding assays, i.e., the protocol provided herein:

[0249] The purpose of this biochemical assay is to measure the ability of the test compound to promote the formation of a ternary complex between nucleotide-loaded Ras isoforms and cyclophilin A; the resulting ternary complex disrupts the binding to BRAF. RBD Binding of the construct inhibits Ras signaling through RAF effectors.

[0250] The culture medium contained 25 mM HEPES pH 7.3, 0.002% Tween 20, 0.1% BSA, 100 mM NaCl and 5 mM MgCl 2 Unlabeled cyclophilin A, His6-K-Ras-GMPPNP (or other Ras variants) and GST-BRAF were added to the assay buffer. RBD The final concentrations of 25 μM, 12.5 nM and 50 nM were combined in a 384-well assay plate. The compounds present in each well of the plate were diluents that were subjected to 10-point 3-fold serial dilutions starting from a final concentration of 30 μM. After incubation for 3 hours at 25°C, a mixture of anti-His Eu-W1024 and anti-GST allophycocyanin was then added to the assay sample wells to reach final concentrations of 10 nM and 50 nM, respectively, and the reaction was incubated for another 1.5 hours. The TR-FRET signal was read on a microplate reader (Ex 320 nm, Em 665 / 615 nm). Compounds that promote K-Ras:RAF complex destruction are identified as compounds that reduce the TR-FRET ratio relative to the DMSO control wells.

[0251] Thus, in some embodiments, the compounds of the invention have an effect on KRAS relative to Ras mutants or relative to wild-type. G12D The compounds of the present invention also exhibit greater selectivity for other RAS mutants disclosed herein or combinations thereof.

[0252] In some embodiments, the compounds of the invention are more potent against one or more specific Ras mutants than against other Ras mutants or wild type compared to compounds known in the art. Methods for measuring this potency are known in the art, such as the pERK assay, a protocol for which is provided in the Examples below. Thus, in some embodiments, the compounds of the invention are more potent against KRAS compared to compounds known in the art. G12D The compounds of the present invention may also exhibit higher potency against other RAS mutants disclosed herein or combinations thereof.

[0253] In some embodiments, the compounds of the invention exhibit a stronger deleterious effect on cell viability against one or more specific Ras mutants relative to other Ras mutants or wild type compared to compounds known in the art. Methods for measuring cell viability are known in the art, such as those described herein. Cell viability assay:

[0254] NOTE - The following protocol describes a procedure for monitoring cell viability of K-Ras mutant cancer cell lines in response to compounds of the invention. Other RAS isoforms may also be employed, but the number of cells seeded will vary depending on the cell line used.

[0255] The purpose of this cell-based assay is to use The effect of test compounds on the proliferation of human Ras cancer cell lines (e.g., NCI-H358 (K-RasG12C), AsPC-1 (K-Ras G12D), and Capan-1 (K-Ras G12V)) over a 5-day treatment period was determined by quantifying the amount of ATP present at the endpoint using the TRIzOL® 2.0 reagent (Promega).

[0256] Cells were seeded at 250 cells / well in 40 μL of growth medium in a 384-well assay plate and incubated at 37°C in a humidified atmosphere with 5% CO. 2 Incubate overnight in a humidified atmosphere of 5% DMSO. On the day of the assay, first, dilute the 10 mM stock solution of the test compound to a 3 mM solution with 100% DMSO. Transfer the well-mixed compound solution (15 μL) to the next batch of wells containing 30 μL of 100% DMSO and repeat until 9 concentrations of 3-fold serial dilutions are prepared (the starting assay concentration is 10 μM). The test compound (132.5 nL) is dispensed directly into the assay plate containing the cells. Each plate is shaken at 300 rpm for 15 seconds, centrifuged and incubated at 37°C in a 5% CO 2 Incubate in a humidified atmosphere for 5 days. On day 5, equilibrate the assay plate and its contents to room temperature for approximately 30 minutes. 2.0 reagent (25 μL) and the plate contents were mixed on an orbital shaker for 2 minutes followed by incubation at room temperature for 10 minutes. Luminescence was measured using a PerkinElmer Enspire. Data were normalized using the formula: (sample signal / average DMSO)*100. Data were fit using a four parameter logistic fit.

[0257] Therefore, in some embodiments, compared with compounds known in the art, the compounds of the present invention have a G12D The compounds of the present invention may also exhibit a significant reduction in cell viability against other RAS mutants disclosed herein or a combination thereof.

[0258] In some embodiments, the compounds of the present invention may exhibit higher metabolic stability, permeability or solubility, or a combination thereof, compared to compounds known in the art. The compounds of the present invention may, for example, exhibit improved acid stability in a simulated gastric fluid stability assay. Methods for measuring these properties are known in the art. Compared to compounds known in the art, the compounds of the present invention may exhibit better Ras cross-linking. Methods for measuring Ras cross-linking are provided herein. In some embodiments, compared to compounds known in the art, the compounds of the present invention may exhibit improvements in any one of the following properties or a combination thereof: selectivity, efficacy, cell viability, metabolic stability, acid stability, cross-linking, permeability or solubility.

[0259] Also provided is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a 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 adenocarcinoma, myelodysplastic syndrome, or squamous cell lung cancer. In some embodiments, the cancer comprises a Ras mutation, such as K-Ras G12D or K-Ras G13D. Other Ras mutations are also described herein.

[0260] Also provided is a method for treating a Ras protein-related disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof.

[0261] Also provided is a method for inhibiting Ras protein in a cell, the method comprising contacting the cell with an effective amount of a compound of the invention or a pharmaceutically acceptable salt thereof. For example, the Ras protein is K-Ras G12D or K-Ras G13D. Other Ras proteins are also described herein. The cell can be a cancer cell, such as a pancreatic cancer cell, a colorectal cancer cell, a non-small cell lung cancer cell, an acute myeloid leukemia cell, a multiple myeloma cell, a thyroid adenocarcinoma cell, a myelodysplastic syndrome cell, or a squamous cell lung cancer cell. Other cancer types are also described herein. The cell can be in vivo or in vitro.

[0262] For compounds of the invention, one stereoisomer may exhibit an inhibitory effect superior to another stereoisomer. For example, one atropisomer may exhibit an inhibitory effect, while another atropisomer may exhibit very little or no inhibitory effect.

[0263] In some embodiments, the methods or uses described herein further include administering additional anticancer therapies. In some embodiments, the additional anticancer therapies are 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, HER2 inhibitors, or combinations thereof. In some embodiments, the additional anticancer therapies are SHP2 inhibitors. Other additional anticancer therapies are also described herein.

[0264] Synthesis method

[0265] The compounds described herein can be prepared from commercially available starting materials or synthesized using known organic, inorganic or enzymatic methods.

[0266] The compounds of the present invention can be prepared in a variety of ways well known to those skilled in the art of organic synthesis. For example, the compounds of the present invention can be synthesized using the methods described in the following schemes, as well as synthetic methods known in synthetic organic chemistry techniques or modifications of the methods understood by those skilled in the art. The methods include, but are not limited to, the methods described in the following schemes.

[0267] The compounds of Table 1 herein are prepared using the methods disclosed herein, or using the methods disclosed herein in combination with the knowledge of those skilled in the art. The compounds of Table 2 can be prepared using the methods disclosed herein, or can be prepared using the methods disclosed herein in combination with the knowledge of those skilled in the art.

[0268] Scheme 1. General synthesis of aziridine-containing macrocycles

[0269]

[0270] As shown in Scheme 1, this class of compounds can be prepared by reacting the appropriate amine (1) with a protected amine-containing carboxylic acid (2) in the presence of standard amide coupling reagents to give 3, followed by deprotection of the amine to produce 4. Aziridine carboxylates (5) are coupled in the presence of standard amide coupling reagents to give the final compounds (6).

[0271] Pharmaceutical compositions and methods of use

[0272] Pharmaceutical compositions and methods of administration

[0273] The compounds involved in the present invention are Ras inhibitors and can be used to treat cancer. Therefore, one embodiment of the present invention provides a pharmaceutical composition containing a compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient, and a method for preparing such a composition using the compound of the present invention.

[0274] As used herein, the term "pharmaceutical composition" refers to a compound, such as a compound of the present invention, or a pharmaceutically acceptable salt thereof, formulated together with a pharmaceutically acceptable excipient.

[0275] In some embodiments, the compound is present in the pharmaceutical composition in an amount of a unit dose suitable for administration in a treatment regimen, and the compound shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, the pharmaceutical composition can be specifically formulated for administration in solid or liquid form, including those suitable for administration in the following ways: oral administration, such as a medicament (aqueous or non-aqueous solution or suspension), a tablet (e.g., intended for buccal, sublingual and systemic absorption), a bolus, a powder, a granule, a paste for application to the tongue; parenteral administration, such as by subcutaneous, intramuscular, intravenous or epidural injection, such as, for example, a sterile solution or suspension, or a sustained release formulation; topical administration, such as a cream, ointment or controlled release patch or spray applied to the skin, lungs or oral cavity; intravaginal or intrarectal administration, such as a pessary, cream or foam; sublingual administration; ocular administration; transdermal administration; or nasal, pulmonary administration and administration to other mucosal surfaces.

[0276] As used herein, "pharmaceutically acceptable excipient" refers to any inactive ingredient (e.g., a vehicle capable of suspending or dissolving an active compound) that is non-toxic and non-inflammatory in a subject. Typical excipients include, for example, antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (pigments), softeners, emulsifiers, fillers (diluents), film formers or coatings, flavoring agents, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweeteners, or water for hydration. Excipients include, but are not limited to, optionally substituted butylated hydroxytoluene (BHT), calcium carbonate, calcium hydrogen phosphate, calcium stearate, cross-linked carboxymethylcellulose, cross-linked polyvinyl pyrrolidone, citric acid, cross-linked polyvinylpyrrolidone, 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 starch), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol. Those skilled in the art are aware of a variety of agents and materials that can be used as excipients. See, e.g., 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 includes at least two different pharmaceutically acceptable excipients.

[0277] Unless clearly stated to the contrary, the compounds described herein, whether or not clearly stated, can be provided or used in the form of salts, for example, in the form of pharmaceutically acceptable salts. As used herein, the term "pharmaceutically acceptable salt" refers to salts of compounds described herein that are suitable for use in contact with human tissues without excessive toxicity, irritation, allergic reactions, etc., and are commensurate with a reasonable benefit / risk ratio within the scope of reasonable medical judgment. 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, (P. H. Stahl and C. G. Wermuth, ed.), Wiley-VCH, 2008. The salts can be prepared in situ during the final separation and purification of the compounds described herein, or separated by reacting the free basic groups with suitable organic acids.

[0278] The compounds of the present invention may have ionizable groups, thereby being able to be prepared into pharmaceutically acceptable salt forms. These salts may be acid addition salts involving inorganic or organic acids, or in the case where the compounds of the present invention are in acid form, the salts may be prepared from inorganic or organic bases. In some embodiments, the compound is prepared as a pharmaceutically acceptable salt form or used in a pharmaceutically acceptable salt form, and the pharmaceutically acceptable salt form is prepared as an addition product of a pharmaceutically acceptable acid or base. Suitable pharmaceutically acceptable acids and bases are well known in the art, 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. Methods for preparing appropriate salts are recognized in the art.

[0279] Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-optionally substituted hydroxy-ethyl Alkanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate, etc. Representative alkali metal 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.

[0280] As used herein, the term "subject" refers to any member of the animal kingdom. In some embodiments, a "subject" refers to a human being at any stage of development. In some embodiments, a "subject" refers to a human patient. In some embodiments, a "subject" refers to a non-human animal. In some embodiments, a non-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, a cow, a primate, or a pig). In some embodiments, subjects include, but are not limited to, mammals, birds, reptiles, amphibians, fish, or insects. In some embodiments, a subject can be a transgenic animal, a genetically engineered animal, or a clone.

[0281] As used herein, the term "dosage form" refers to a physical discrete unit of a compound (e.g., a compound of the invention) for administration to a subject. Each unit contains a predetermined amount of compound. In some embodiments, the amount is the amount of a unit dose (or its entire portion) suitable for administration according to a dosage regimen, which is determined to be associated with a desired or beneficial result when administered to a relevant population (i.e., according to a therapeutic dosage regimen). It will be appreciated by those skilled in the art that the total amount of the therapeutic composition or compound administered to a particular subject is determined by one or more attending physicians and may involve the administration of a variety of dosage forms.

[0282] As used herein, the term "dosage regimen" refers to a group of unit doses (typically more than one unit dose) administered individually to a subject, the unit doses typically being spaced apart for a certain period of time. In some embodiments, a given therapeutic compound (e.g., a compound of the present invention) has a recommended dosage regimen that may involve one or more doses. In some embodiments, the dosage regimen comprises multiple doses, each of which is spaced apart from each other by a period of the same length; in some embodiments, the dosage regimen comprises multiple doses and at least two different time periods separate individual doses. In some embodiments, all doses within the dosage regimen are the same unit dose amount. In some embodiments, different doses within the dosage regimen are different amounts. In some embodiments, the dosage regimen comprises a first dose of the amount of a first dose, followed by one or more additional doses of the amount of a second dose different from the amount of the first dose. In some embodiments, the dosage regimen comprises a first dose of the amount of a first dose, followed by one or more additional doses of the amount of a second dose identical to the amount of the first dose. In some embodiments, the dosage regimen is associated with desired or beneficial results when administered to a relevant population (i.e., a therapeutic dosage regimen).

[0283] A "therapeutic regimen" refers to a dosage regimen whose administration is associated with desired or beneficial therapeutic results in the relevant population.

[0284] The term "treatment" ("treatment / treat / treating") in its broadest sense refers to any administration of a substance (e.g., a compound of the present invention) that partially or completely alleviates, ameliorates, alleviates, inhibits one or more symptoms, features, or causes of a particular disease, disorder, or condition; delays its onset; reduces its severity; or reduces its occurrence. In some embodiments, such treatments may be administered to subjects who do not show signs of the relevant disease, disorder, or condition or to subjects who only show early signs of the disease, disorder, or condition. Alternatively or additionally, in some embodiments, treatment may be administered to subjects who show one or more determined signs of the relevant disease, disorder, or condition. In some embodiments, treatment can be used for subjects diagnosed with a relevant disease, disorder, or condition. In some embodiments, treatment can be used for subjects known to have one or more susceptibility factors that are statistically associated with an increased risk of developing a relevant disease, disorder, or condition.

[0285] The term "therapeutically effective amount" means an amount sufficient to treat a disease, disorder or illness when administered to a population suffering from or susceptible to a disease, disorder or illness according to a therapeutic dosage regimen. In some embodiments, a therapeutically effective amount is an amount that reduces the incidence or severity of one or more symptoms of the disease, disorder or illness or delays its onset. It will be understood by those skilled in the art that the term "therapeutically effective amount" does not actually require the desired successful treatment to be achieved in a particular individual. In fact, a therapeutically effective amount can be an amount that provides a specific desired pharmacological response in a considerable number of subjects when administered to a patient in need of the treatment. It is particularly important to understand that a particular subject may actually be "refractory" to a "therapeutically effective amount". In some embodiments, the therapeutically effective amount mentioned may refer to an amount measured in one or more specific tissues (e.g., tissues affected by a disease, disorder or illness) or fluids (e.g., blood, saliva, serum, sweat, tears, urine). It will be understood by those skilled in the art that in some embodiments, a therapeutically effective amount may be formulated as a single dose or administered in a single dose. In some embodiments, a therapeutically effective amount may be formulated as multiple doses, for example, as part of a dosing regimen, or administered as multiple doses.

[0286] For use as a treatment for a subject, the compounds of the present invention or their pharmaceutically acceptable salts can be formulated as a pharmaceutical composition or a veterinary composition. Depending on the subject to be treated, the mode of administration, and the desired type of treatment, such as prevention, control, or treatment, the compound or its pharmaceutically acceptable salt is formulated in a manner consistent with the parameters. An overview of such techniques can be found in Remington: The Science and Practice of Pharmacy, 21st edition, Lippincott Williams & Wilkins, (2005); and Encyclopedia of Ph armaceutical Technology, edited by J. Swarbrick and JC Boylan, 1988-1999, Marcel Dekker, New York, each of which is incorporated herein by reference.

[0287] The composition can be prepared according to conventional mixing, granulation or coating methods, respectively, and the pharmaceutical composition of the present invention can 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 its pharmaceutically acceptable salt by weight or volume. In some embodiments, the compound described herein or its pharmaceutically acceptable salt is present in an amount by weight that can account for 1-95% of the total amount of the composition, such as the pharmaceutical composition.

[0288] The composition can be provided in a dosage form suitable for the following administration: intra-articular, oral, parenteral (e.g., intravenous, intramuscular), rectal, skin, subcutaneous, surface, transdermal, sublingual, nasal, vaginal, intracapsular, intraurethral, ​​intrathecal, epidural, ear or eye administration, or by injection, inhalation or direct contact with the nose, urogenital, genital or oral mucosa. Therefore, the pharmaceutical composition can be in the form of, for example, tablets, capsules, pills, powders, granules, suspensions, emulsions, solutions, gels (including hydrogels), pastes, ointments, creams, plasters, syrups, osmotic delivery devices, suppositories, enemas, injections, implants, sprays, preparations suitable for iontophoresis delivery or aerosols. The composition can be prepared according to conventional medical practice.

[0289] As used herein, the term "administering" refers to administering a composition (e.g., a compound or a formulation comprising a compound as described herein) to a subject or system. Administration to an animal subject (e.g., to a human) can be performed by any appropriate route. For example, in some embodiments, administration can be bronchial (including by bronchial instillation), buccal, enteral, intradermal, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intracapsular, transmucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal, or vitreous administration.

[0290] The formulation can be prepared in a manner suitable for systemic administration or topical or local administration. Systemic formulations include formulations designed for injection (e.g., intramuscular, intravenous or subcutaneous injection) or can be prepared for transdermal, transmucosal or oral administration. The formulation will generally include a diluent, and in some cases, adjuvants, buffers, preservatives, etc. The compound or its pharmaceutically acceptable salt can also be administered in a liposome composition or in a microemulsion form.

[0291] For injection, the formulation can be prepared in conventional forms, such as liquid solutions or suspensions, or solid forms suitable for preparation as solutions or suspensions in liquids prior to injection, or in the form of emulsions. Suitable excipients include, for example, water, saline, dextrose, glycerol, etc. These compositions may also contain a certain amount of non-toxic auxiliary substances, such as wetting agents or emulsifiers, pH buffers, etc., such as sodium acetate, sorbitan monolaurate, etc.

[0292] Various sustained-release drug systems have also been designed. See, for example, U.S. Pat. No. 5,624,677.

[0293] 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 invention or their pharmaceutically acceptable salts. It will be appreciated in the art that suitable forms include syrups, capsules, and tablets.

[0294] Each compound described herein or its pharmaceutically acceptable salt can be formulated in a variety of ways known in the art. For example, the first agent and the second agent in the combination therapy can be formulated together or separately. Other modes of combination therapy are also described herein.

[0295] Individual or separately formulated medicaments can be packaged together in the form of a kit. Non-limiting examples include, but are not limited to, kits containing, for example, two pills, one pill and a powder, a suppository or a liquid in a vial, two surface creams, etc. The kit may include optional components that help to administer a unit dose to a subject, such as vials, syringes for injection, custom IV delivery systems, inhalers, etc., for reconstitution of the powder form. In addition, the unit dose kit may contain instructions for the preparation and administration of the composition. The kit may be manufactured as a disposable unit dose for a subject, for multiple uses for a specific subject (at a constant dose, or where the efficacy of an individual compound or its pharmaceutically acceptable salt may vary with treatment progress); or the kit may contain multiple doses suitable for administering multiple subjects ("integral packaging"). The kit components may be assembled in cartons, blister packs, bottles, tubes, etc.

[0296] Formulations for oral use include tablets containing the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients. The excipients may be, for example, inert diluents or fillers (e.g., sucrose, sorbitol, sugar, mannitol, microcrystalline cellulose, starch including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate or sodium phosphate); granulating and disintegrants (e.g., cellulose derivatives including microcrystalline cellulose, starch including potato starch, cross-linked sodium carboxymethyl cellulose, alginates or alginic acid); binders (e.g., sucrose, glucose, sorbitol, gum arabic, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, sodium carboxymethyl cellulose, methyl cellulose, optionally substituted hydroxypropyl methyl cellulose, ethyl cellulose, polyvinyl pyrrolidone or polyethylene glycol); and lubricants, glidants and anti-adherents (e.g., magnesium stearate, zinc stearate, stearic acid, silicon dioxide, hydrogenated vegetable oils or talc). Other pharmaceutically acceptable excipients may be colorants, flavoring agents, plasticizers, humectants, buffers, and the like.

[0297] Two or more compounds can be mixed together in tablets, capsules or other vehicles, or can be separated. In one embodiment, the first compound is contained on the inside of the tablet, and the second compound is on the outside, thereby making the vast majority of the second compound release before the first compound releases.

[0298] Formulations for oral use may also be provided in the form of chewable tablets, or in the form of hard 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 in the form of soft gelatin capsules in which the active ingredient is mixed with water or an oil medium, e.g. peanut oil, liquid paraffin or olive oil. Powders, granules and granules may be prepared using the ingredients mentioned above for tablets and capsules in a conventional manner, using, for example, a mixer, a fluid bed apparatus or a spray drying device.

[0299] Dissolution or diffusion controlled release can be achieved by appropriately coating tablets, capsules, granules or granular formulations of the compound, or by incorporating the compound or its pharmaceutically acceptable salt into a suitable matrix. The controlled release coating may include one or more of the above-mentioned coating materials, such as shellac, beeswax, glycowax, castor wax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryl distearate, glyceryl palmitostearate, ethylcellulose, acrylic resin, dl-polylactic acid, cellulose acetate butyrate, polyvinyl chloride, polyvinyl acetate, vinyl pyrrolidone, polyethylene, polymethacrylate, methyl methacrylate, 2-optionally substituted hydroxymethylacrylate, 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, tristearin, methyl acrylate-methyl methacrylate, polyvinyl chloride, polyethylene or halogenated fluorocarbons.

[0300] Liquid forms for oral administration that may incorporate the compounds of the invention or their pharmaceutically acceptable salts and compositions 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.

[0301] In general, when administered to humans, the oral dosage of any compound of the present invention or its pharmaceutically acceptable salt will depend on the properties of the compound, and can be easily determined by those skilled in the art. Dosage can 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 derived therefrom. In some embodiments, the daily dosage range for oral administration can be, for example, within the range of about 0.001 mg to about 2000 mg per kilogram of human body weight, administered in a single dose or divided dose. On the other hand, in some cases, it may be necessary to use dosages outside these limits.

[0302] In some embodiments, the pharmaceutical composition may further comprise an additional compound having antiproliferative activity. Depending on the mode of administration, the compound or its pharmaceutically acceptable salt will be formulated into a suitable composition for delivery. Each compound or its pharmaceutically acceptable salt in the combination therapy can be formulated in a variety of ways known in the art. For example, the first agent and the second agent in the combination therapy can be formulated together or separately. Desirably, the first agent and the second agent are formulated together so that the agents are administered simultaneously or nearly simultaneously.

[0303] It should be understood that 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 with one or more other desired therapeutic agents or medical procedures or administered at the same time, before or after the administration of the one or more other desired therapeutic agents or medical procedures. The specific combination of each therapy (therapeutic agent or procedure) used in the combination regimen should take into account the compatibility of the desired therapeutic agent or procedure with the desired therapeutic effect to be achieved. It should also be understood that the therapies employed can achieve the desired effect for the same condition, or that these therapies can achieve different effects (e.g., control any adverse effects).

[0304] As described herein, administration of each drug in the combination therapy can be independently one to four times daily for one day to one year, or even for the life of the subject. Chronic (long-term) administration may also be applicable.

[0305] How to use

[0306] In some embodiments, the present invention discloses a method of treating a disease or condition characterized by aberrant Ras activity caused by a Ras mutant. In some embodiments, the disease or condition is cancer.

[0307] Therefore, a method for treating cancer in a subject in need is also provided, the method comprising administering to the subject a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising such a compound or salt. In some embodiments, the cancer is colorectal cancer, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, appendix cancer, melanoma, acute myeloid leukemia, small intestine cancer, ampullary cancer, germ cell cancer, cervical cancer, cancer of unknown primary site, endometrial cancer, esophagogastric cancer, GI neuroendocrine cancer, ovarian cancer, sex cord stromal tumor cancer, hepatobiliary cancer, or bladder cancer. In some embodiments, the cancer is appendix cancer, endometrial cancer, or melanoma. A method for treating a Ras protein-related disorder in a subject in need is also provided, the method comprising administering to the subject a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising such a compound or salt.

[0308] In some embodiments, the compounds of the present invention or their pharmaceutically acceptable salts, pharmaceutical compositions comprising such compounds or salts, and methods provided herein can be used to treat a variety of cancers, including tumors, such as lung cancer, prostate cancer, breast cancer, brain cancer, skin cancer, cervical cancer, testicular cancer, etc. More specifically, cancers that can be treated by the compounds of the present invention or their salts, pharmaceutical compositions comprising such compounds or salts, and methods include, but are not limited to, the following tumor types: astrocytes, breast, cervical, colorectal, endometrial, esophageal, gastric, head and neck, hepatocellular, laryngeal, lung, oral, ovarian, prostate, and thyroid carcinomas and sarcomas. Other cancers include, for example:

[0309] Heart, such as sarcomas (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma, and teratoma;

[0310] Lung, for example: bronchogenic carcinoma (squamous cell lung cancer, undifferentiated small cell lung cancer, undifferentiated large cell lung cancer, lung adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, enchondromatous hamartoma, mesothelioma;

[0311] Gastrointestinal, for example: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, vipoma), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma);

[0312] Urogenital tract, for example: kidney (adenocarcinoma, Wilm's tumor (Nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatoid tumor, lipoma);

[0313] Liver, for example: hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma;

[0314] Biliary tract, such as gallbladder cancer, ampullary cancer, and bile duct cancer;

[0315] Bone, such as osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticular cell sarcoma), multiple myeloma, malignant giant cell tumor, chordoma, osteochondroma (osteocartilaginous exostosis), benign enchondroma, chondroblastoma, chondromyxoid fibroma, osteoid osteoma, and giant cell tumor;

[0316] Nervous system, for example: skull (osteomas, hemangiomas, granulomas, xanthomas, osteitis deformans), meninges (meningiomas, meningosarcomas, gliomatosis), brain (astrocytomas, medulloblastomas, gliomas, ependymomas, germ cell tumors (pinealomas), glioblastoma multiforme, oligodendrogliomas, schwannomas, retinoblastomas, congenital tumors), spinal neurofibromas, neurofibromatosis type 1, meningiomas, gliomas, sarcomas);

[0317] Gynecology, for example: uterus (endometrial cancer, uterine cancer, endometrial cancer), cervix (cervical cancer, cervical precancerous atypical hyperplasia), ovary (ovarian cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa-theca cell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma), fallopian tube (carcinoma);

[0318] Hematopoietic system, for example: blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative diseases (such as myelofibrosis and myeloproliferative neoplasms), multiple myeloma, myelodysplastic syndromes), Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma);

[0319] Skin, such as: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, nevus dysplasia, lipoma, hemangioma, dermatofibroma, keloid, psoriasis; and

[0320] Adrenal glands, e.g. neuroblastoma.

[0321] In some embodiments, the Ras protein is wild-type (Ras WT ). Therefore, in some embodiments, the compounds of the invention are used to treat patients with Ras WT (e.g. K-Ras WT 、H-Ras WT N-RasWT ) of a patient with cancer. In some embodiments, the Ras protein is a Ras amplified (eg, K-Ras amp ). Therefore, in some embodiments, the compounds of the invention are used to treat patients with Ras amp (K-Ras amp 、H-Ras amp N-Ras amp ) in a method of treating a patient with cancer. In some embodiments, the cancer comprises a Ras mutation, such as a Ras mutation described herein. In some embodiments, the mutation is selected from:

[0322] (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;

[0323] (b) the following H-Ras mutants: 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;

[0324] (c) the following N-Ras mutants: 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;

[0325] or a combination of any of the foregoing. In some embodiments, the compounds inhibit K-Ras G12C and K-Ras G12D. In some embodiments, the compounds inhibit both K-Ras G12D and K-Ras G13D. In some embodiments, the compounds inhibit both K-Ras G12V and K-Ras G12S. In some embodiments, the compounds of the present invention inhibit Ras WT and one or more additional Ras mutations (e.g., K, H, or N-Ras WT In some embodiments, the compounds of the invention inhibit Ras ampand one or more additional Ras mutations (e.g., K-, H-, or N-Ras amp and K-RasG12D).

[0326] Methods for detecting Ras mutations are known in the art. Such methods include, but are not limited to, direct sequencing and using highly sensitive diagnostic assays (using CE-IVD marking), such as Domagala et al., Pol J Pathol 3: 145-164 (2012), which are incorporated herein by reference in their entirety, including TheraScreenPCR; AmoyDx; PNAClamp; RealQuality; EntroGen; LightMix; StripAssay; HybcellplexA; Devyser; Surveyor; Cobas; and TheraScreenPyro. See also, for example, WO2020 / 106640.

[0327] In some embodiments, the cancer is non-small cell lung cancer and the Ras mutation comprises a K-Ras mutation, such as K-RasG12D. In some embodiments, the cancer is colorectal cancer and the Ras mutation comprises a K-Ras mutation, such as K-RasG12D. In some embodiments, the cancer is pancreatic cancer and the Ras mutation comprises a K-Ras mutation, such as K-Ras G12D. In some embodiments, the cancer is pancreatic cancer and the Ras mutation comprises an N-Ras mutation, such as N-Ras G12D. In any of the foregoing, if not specifically stated, the compound may also inhibit Ras WT (e.g. K-, H- or N-Ras WT ) or Ras amp (e.g. K-, H- or N-Ras amp ).

[0328] A method for inhibiting Ras protein in a cell is also provided, the method comprising contacting the cell with an effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof. The compound or a pharmaceutically acceptable salt thereof can inhibit more than one type of Ras protein in a cell. A method for inhibiting RAF-Ras binding is also provided, the method comprising contacting the cell with an effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof. The cell may be a cancer cell. The cancer cell may belong to any type of cancer described herein. The cell may be in vivo or in vitro.

[0329] Combination therapy

[0330] The methods of the present invention may include the compounds of the present invention used alone or in combination with one or more additional therapies (e.g., non-drug treatments or therapeutic agents). When administered alone, the dosage of one or more of the additional therapies (e.g., non-drug treatments or therapeutic agents) may be reduced relative to the standard dosage. For example, the dosage may be determined empirically based on drug combinations and permutations, or may be inferred by isoradiometric determinations (e.g., Black et al., Neurology 65: S3-S6 (2005)).

[0331] The compounds of the invention may be administered before, after, or simultaneously with the administration of one or more of the additional therapies. When combined, the dose of the compounds of the invention and the dose of the one or more additional therapies (e.g., non-drug therapies or therapeutic agents) provide a therapeutic effect (e.g., a synergistic or additive therapeutic effect). The compounds of the invention and additional therapies, such as anticancer agents, may be administered together, such as in the form of a single pharmaceutical composition, or separately, and when administered separately, the administration may occur simultaneously or sequentially. Such sequential administration may be close or distant in time.

[0332] In some embodiments, the additional therapy is the administration of a side effect limiting agent (e.g., an agent intended to reduce the occurrence or severity of a therapeutic side effect). For example, in some embodiments, the compounds of the invention may also be used in combination with a therapeutic agent for treating nausea. Examples of agents that can be used to treat nausea include: granisetron, metoclopramide, ondansetron, and prochlorperazine, or a pharmaceutically acceptable salt thereof.

[0333] In some embodiments, the one or more additional therapies include non-drug therapy (e.g., surgery or radiotherapy). In some embodiments, the one or more additional therapies include therapeutic agents (e.g., compounds or biological agents as anti-angiogenic agents, signal transduction inhibitors, anti-proliferative agents, glycolysis inhibitors, or autophagy inhibitors). In some embodiments, the one or more additional therapies include non-drug therapy (e.g., surgery or radiotherapy) and therapeutic agents (e.g., compounds or biological agents as anti-angiogenic agents, signal transduction inhibitors, anti-proliferative agents, glycolysis inhibitors, or autophagy inhibitors). In other embodiments, the one or more additional therapies include two therapeutic agents. In yet other embodiments, the one or more additional therapies include three therapeutic agents. In some embodiments, the one or more additional therapies include four or more therapeutic agents.

[0334] In this Combination Therapies section, all references are incorporated by reference with respect to the described agents, or pharmaceutically acceptable salts, solvates, isomers (eg, stereoisomers), prodrugs, or tautomers thereof, whether or not expressly stated as such.

[0335] Non-drug treatments

[0336] Examples of non-drug treatments include, but are not limited to, radiation therapy, cryotherapy, hyperthermia, surgery (eg, surgical removal of tumor tissue), and adoptive T-cell transfer (ACT) therapy.

[0337] In some embodiments, the compounds of the present invention can be used as postoperative adjuvant therapy. In some embodiments, the compounds of the present invention can be used as preoperative neoadjuvant therapy.

[0338] Radiation therapy can be used to inhibit abnormal cell growth or treat hyperproliferative disorders such as cancer in a subject, such as a mammal (e.g., a human). Techniques for administering radiation therapy are known in the art. Radiation therapy can be administered by one or a combination of methods, including but not limited to external beam therapy, internal radiation therapy, implant radiation, stereotactic radiosurgery, systemic radiation therapy, radiotherapy, and long-term or short-term implantation of brachytherapy. As used herein, the term "brachytherapy" refers to radiation therapy delivered by spatially confined radioactive materials inserted at or near a tumor or other proliferative tissue disease site in the body. This term is intended, but is not limited to, to include 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 conditioners of the present invention include solids and liquids. As non-limiting examples, the radiation source can be a radionuclide, such as 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 radiation or other therapeutic rays. The radioactive material can also be a fluid made from any radionuclide solution, such as a solution of I-125 or I-131, or the radioactive fluid can be prepared using a slurry of a suitable fluid containing small particles of solid radionuclides such as Au-198 or Y-90. In addition, the radionuclide can be embedded in a gel or radioactive microspheres.

[0339] In some embodiments, the compounds of the present invention can sensitize abnormal cells to radiotherapy to achieve the purpose of killing or inhibiting the growth of such cells. Therefore, the present invention further relates to a method for sensitizing abnormal cells in a mammal to radiotherapy, the method comprising administering to the mammal an amount of the compounds of the present invention, the amount of which effectively sensitizes abnormal cells to radiotherapy. The amount of the compound in the method can be determined according to the method for determining the effective amount of the compound described herein. In some embodiments, the compounds of the present invention can be used as an adjuvant therapy after radiotherapy or as a neoadjuvant therapy before radiotherapy.

[0340] In some embodiments, the non-drug treatment is T cell adoptive transfer (ACT) therapy. In some embodiments, the T cell is an activated T cell. The T cell can be modified to express a chimeric antigen receptor (CAR). CAR-modified T (CAR-T) cells can be produced by any method known in the art. For example, CAR-T cells can be produced by introducing a suitable expression vector encoding CAR into T cells. Before T cells are amplified and genetically modified, the source of T cells is obtained from the subject. T cells can be obtained from a variety of sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, umbilical cord blood, thymus tissue, tissue from the site of infection, ascites, pleural effusion, spleen tissue and tumor. In certain embodiments of the present invention, a variety of T cell lines available in the art can be used. In some embodiments, the T cell is an autologous T cell. Before or after the T cells are genetically modified to express a desired protein (e.g., CAR), these T cells can generally be activated and expanded using methods such as described in, for example, the following U.S. Patents: 6,352,694; 6,534,055; 6,905,680; 6,692,964; 5,858,358; 6,887,466; 6,905,681; 7,144,575; 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.

[0341] Therapeutic agents

[0342] The therapeutic agent can be a compound used to treat cancer or its related symptoms. The compounds of the present invention can be combined with a second therapeutic agent, a third therapeutic agent, or a fourth therapeutic agent, or more therapeutic agents. The compounds of the present invention can be combined with one or more therapeutic agents and one or more non-drug therapies.

[0343] For example, the therapeutic agent can be a steroid. Steroids are known in the art. Therefore, in some embodiments, the one or more additional therapies include steroids. Suitable steroids may include, but are not limited to, 21-acetoxypregnenolone, alclometasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, clocortolone, cloprednol, corticosterone, cortisol, sone, cortivazol, deflazacort, desonide, desoximetasone, dexamethasone, diflorasone, diflucortolone, difuprednate, enoxolone, fluazacort, fiucloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinonide, fluocortinbutyl, fluocortolone, fluorometholone, fluperolone acetate, fluprednidene acetateacetate), fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halcinonide, halobetasol propionate, halometasone, hydrocortisone, loteprednoletabonate, mazipredone, medrysone, meprednisone, methylprednisolone , mometasone furoate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethylaminoacetate, prednisolone sodium phosphate, prednisone, prednival, prednylidene, rimexolone, tixocortol, triamcinolone, triamcinoloneacetonide, triamcinolonebenetonide, triamcinolone hexacetonide, and salts or derivatives thereof.

[0344] Other examples of therapeutic agents that can be used in combination therapy with the compounds of the invention include compounds described in U.S. Pat. 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 8,623,885, as well as 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.

[0345] The therapeutic agent can be a biological agent (e.g., a cytokine (e.g., interferon or interleukin, such as IL-2)) for treating cancer or its associated symptoms. Biological agents are known in the art. In some embodiments, the biological agent is an immunoglobulin-based biological agent, such as a monoclonal antibody (e.g., a humanized antibody, a fully human antibody, an Fc fusion protein, or a functional fragment thereof) that excites the target to stimulate an anti-cancer response or antagonizes an antigen that is important for cancer. Antibody-drug conjugates are also included.

[0346] The therapeutic agent can be a T cell checkpoint inhibitor. These 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 can be, for example, a humanized or fully human antibody. In some embodiments, the checkpoint inhibitor is a fusion protein, such as an Fc-receptor fusion protein. In some embodiments, the checkpoint inhibitor is an agent that interacts with the checkpoint protein, such as an antibody. In some embodiments, the checkpoint inhibitor is an agent that interacts with the ligand of the checkpoint protein, such as an antibody. In some embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or a small molecule inhibitor) of CTLA-4 (e.g., an anti-CTLA-4 antibody or a fusion protein). In some embodiments, the checkpoint inhibitor is a PD-1 inhibitor or antagonist (e.g., an inhibitory antibody or a small molecule inhibitor). In some embodiments, the checkpoint inhibitor is a PD-L1 inhibitor or antagonist (e.g., an inhibitory antibody or a small molecule inhibitor). In some embodiments, the checkpoint inhibitor is a PD-L2 inhibitor or antagonist (e.g., an inhibitory antibody or an Fc fusion or a small molecule inhibitor) (e.g., a PD-L2 / Ig fusion protein). In some embodiments, the checkpoint inhibitor is an inhibitor or antagonist (e.g., inhibitory antibody or small molecule inhibitor) of B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK 1, CHK2, A2aR, B-7 family ligand, or a combination thereof. In some embodiments, the checkpoint inhibitor is pembrolizumab, nivolumab, PDR001 (NVS), REGN2810 (Sanofi / Regeneron), PD-L1 antibodies such as avelumab, durvalumab, atezolizumab, pidilizumab, JNJ-63723283 (JNJ), BGB-A317 (BeiGene & Celgene), or P Checkpoint inhibitors disclosed in Reusser, M. et al. (2015) Nat. Rev. Neurol., including but not limited to ipilimumab, tremelimumab, nivolumab, pembrolizumab, AMP224, AMP514 / MEDI0680, BMS936559, MED14736, MPDL3280A, MSB0010718C, BMS986016, IMP321, lirilumab, IPH2101, 1-7F9 and KW-6002.

[0347] The therapeutic agent can be an anti-TIGIT antibody, such as MBSA43, BMS-986207, MK-7684, COM902, AB154, MTIG7192A, or OMP-313M32 (etigilimab). Other anti-TIGIT antibodies are known in the art.

[0348] The therapeutic agent can be an agent for treating cancer or its related symptoms (e.g., a cytotoxic agent, a non-peptide small molecule, or other compounds that can be used to treat cancer or its related symptoms, collectively referred to as "anti-cancer agents"). Anti-cancer agents can be, for example, chemotherapeutic agents or targeted therapeutic agents. These agents are known in the art.

[0349] Anticancer agents include mitotic inhibitors, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, alkylating agents, antimetabolites, folic acid analogs, pyrimidine analogs, purine analogs and related inhibitors, vinca alkaloids, epipodophyllotoxins, antibiotics, L-asparaginase, topoisomerase inhibitors, interferons, platinum coordination complexes, anthracenedione substituted urea, methylhydrazine derivatives, adrenocortical inhibitors, adrenocortical steroids, progestins, estrogens, antiestrogens, androgens, antiandrogens, and gonadotropin-releasing hormone analogs. Other anticancer agents include leucovorin (LV), irinotecan, oxaliplatin, capecitabine, paclitaxel, and doxetaxel. In some embodiments, the one or more additional therapies include two or more anticancer agents. The two or more anticancer agents can be used in a mixture for combined administration or separate administration. Suitable dosing regimens for combined anticancer agents are known in the art and described in, for example, Saltz et al., Proc.Am.Soc.Clin.Oncol.18:233a (1999), and Douillard et al., Lancet 355(9209):1041-1047 (2000).

[0350] Other non-limiting examples of anticancer agents include (Imatinib Mesylate); (carfilzomib); (bortezomib); Casodex (bicalutamide); (gefitinib); alkylating agents, such as thiotepa and cyclophosphamide; alkyl sulfonates, such as busulfan, improsulfan, and piposulfan; aziridines, such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines and methylmelamines, including altretamine, triethylenemelamine, triethylenephosphamide, triethylenethiophosphamide, and trimethylolmelamine; polyacetyl groups (especially bullatacin) cin and bullatacinone); camptothecin (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its synthetic analogues adozelesin, carzelesin and bizelesin); cryptophycin (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including its synthetic analogues , KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin A; spongistatin; nitrogen mustards, such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine hydrochloride, and mechlorethamine oxide hydrochloride. neoxide hydrochloride), melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracilmustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine and ranimustine;Antibiotics, such as enediyne antibiotics (e.g., calicheamicins, such as calicheamicin γll and calicheamicin ωll (see, e.g., Agnew, Chem. Intl. Ed Engl. 33:183-186 (1994)); dynemicins, such as dynemicin A; bisphosphonates, such as clodronate; esperamicin; neocarzinostatin chromophores and related chromophores; enediyne antibiotic chromophores, aclacinomycin, actinomycin actinomycin, authramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, caminomycin, carminomycin, carzinophilin, chromomycins, dactinomycin, daunorubicin, ditoxin detorubicin, 6-diazo-5-oxo-L-norleucine, adriamycin (doxorubicin), N-morpholino doxorubicin, cyanomorpholino doxorubicin, (2-pyrrolino-doxorubicin, deoxydoxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin (such as mitomycin C), mycophenolic acid, nogalamycin, olivetomycin ivomycin, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites, such as methotrexate and 5-fluorouracil (5-FU);Folic acid analogs, such as denopterin, pteropterin, and trimetrexate; purine analogs, such as fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine; pyrimidine analogs, such as ancitabine, azacitidine, 6-thioazolopyrimidine, carmofur, cytarabine, dideoxyuridine, and doxifluridine. ne), enocitabine, and floxuridine; androgens, such as caprotestosterone, dromostanolone propionate, epitiostanol, mepitiostane, and testolactone; antiadreners, such as aminoglutethimide, mitotane, and trilostane; folic acid supplements, such as folinic acid, acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate; epsilon othilone, such as epsilon B; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansinoids, such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; losoxantrone;Podophyllinic acid; (2-ethylhydrazide; procarbazine; Polysaccharide complex (JHS Natural Products, Eugene, OR); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid acid; triaziquone; 2,2',2"-trichlorotriethylamine; trichothecenes, such as T-2 toxin, verracurin A, roridin A, and anguidine; urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, such as (paclitaxel), (Nanoparticulate formulations of paclitaxel without polyoxyethylene hydrogenated castor oil and albumin engineered) and (docetaxel); chloranbucil; tamoxifen (Nolvadex TM ); raloxifene; aromatase inhibiting 4(5)-imidazoles; 4-hydroxytamoxifen; trioxifene; keoxifene; LY117018; onapristone; toremifene Flutamide, nilutamide, bicalutamide, leuprolide, goserelin; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; platinum coordination complexes, such as cisplatin, oxaliplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; (vinorelbine); novantrone; teniposide; edatrexate; daunomycin; aminopterin; ibandronate; irinotecan (e.g., CPT-11); topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoids, such as retinoic acid; esperamicins; capecitabine (e.g., ); and pharmaceutically acceptable salts of any one of the foregoing.

[0351] Additional non-limiting examples of anticancer agents include trastuzumab Bevacizumab Cetuximab (cetuximabv) Rituximab ABVD, avicine, abagovomab, acridinecarboxamide, adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, alpharadin, alvocidib, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone, amonafide, anthracenedione edione), anti-CD22 immunotoxins, antineoplastic agents (e.g., cell cycle nonspecific antineoplastic agents and other antineoplastic agents described herein), anti-tumorigenic herbs, apaziquone, atiprimod, azathioprine, belotecan, bendamustine, BIBW2992, biricodar, brostallicin, bryostatin, buthionine sulfoximine,sulfoximine), CBV (chemotherapy), calyculin, dichloroacetic acid, discordrin, elsamitrucin, enocitabine, eribulin, exatecan, exisulind, ferruginol, forodesine, fosfestrol, ICE chemotherapy regimen, IT-101, imexon, imiquimod, indolocarbazole, irofulven, l-quinazolam, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine, oxadiazine aniquidar, larotaxel, lenalidomide, lucanthone, lurtotecan, mafosfamide, mitozolomide, nafoxidine, nedaplatin, olaparib, ortataxel, PAC-1, papaya, pixantrone, proteasome inhibitors, rebeccamycin, resiquimod, rubitecan, SN-38, salinosporamide A), sapacitabine, Stanford V, swainsonine, talaporfin, tariquidar, tegafur-uracil, temodar, tesetaxel, triplatintetranitrate, tris(2-chloroethyl)amine, troxacitabine, uramustine, vadimezan, vinflunine, ZD6126, and zosuquidar.

[0352] Other non-limiting examples of anticancer agents include natural products such as vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine), epipodophyllotoxins (e.g., etoposide and teniposide), antibiotics (e.g., dactinomycin / actinomycin D), daunomycin, and idarubicin), anthracyclines, mitoxantrone, bleomycins, plicamycin (mithramycin), mitomycins, enzymes (e.g., L-asparaginase, which systemically metabolizes L-asparagine and removes cells that cannot synthesize asparagine themselves), antiplatelet agents, antiproliferative / antimitotic alkylating agents such as nitrogen mustards (e.g., methylchlorambucil, cyclophosphamide, and the analogs melphalan and chlorambucil), ethyleneimine, and methylmelamines (e.g., hexamethyl melamine and thiotepa), CDK inhibitors (e.g., CDK4 / 6 inhibitors such as abemaciclib, ribociclib, palbociclib, seliciclib, UCN-01, P1446A-05, PD-0332991, dinaciclib, P27-00, AT-7519, RGB286638, and SCH727965), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) and the like,and streptozocin), trazenes-dacarbazinine (DTIC), antiproliferative / antimitotic antimetabolites (such as folic acid analogs), pyrimidine analogs (such as fluorouracil, azathiouracil and cytarabine), purine analogs and related inhibitors (such as mercaptopurine, thioguanine, pentostatin and 2-chlorodeoxyadenosine), aromatase inhibitors (such as anastrozole, exemestane and letrozole) and platinum coordination complexes (such as cisplatin and carboplatin), procarbazine, hydroxyurea, mitotane, amine glutethimide, histone deacetylase (HDAC) inhibitors (such as trichostatin, sodium butyrate, apicidan, suberoylanilide hydroamic acid acid, vorinostat, belinostat, LBH 589, romidepsin, ACY-1215, and panobinostat), mTOR inhibitors (e.g., vistusertib, temsirolimus, everolimus, ridaforolimus, and sirolimus), KSP (Eg5) inhibitors (e.g., Array520), DNA binding agents (e.g., ), PI3K inhibitors such as PI3Kδ inhibitors (e.g. GS-1101 and TGR-1202), PI3Kδ and γ inhibitors (e.g. CAL-130), copanlisib, alpelisib and idelalisib; multikinase inhibitors (e.g. TG02 and sorafenib), hormones (e.g. estrogens and hormone agonists such as luteinizing hormone-releasing hormone (LHRH) agonists (e.g. goserelin, leuprolide and triptorelin), BAFF neutralizing antibodies (e.g. LY2127399), IKK inhibitors, p38 MAPK inhibitors, anti-IL-6 (e.g. CNT0328), telomerase inhibitors (e.g. GRN 163L), Aurora kinase inhibitors (e.g. MLN8237), cell surface monoclonal antibodies (e.g. anti-CD38 (HUMAX-CD38)), anti-CS1 (e.g. elotuzumab), HSP90 inhibitors (e.g. 17AAG and KOS 953), P13K / Akt inhibitors (e.g. perifosine), Akt inhibitors (e.g. GSK-2141795), PKC inhibitors (e.g. enzastaurin), FTIs (e.g. Zarnestra TM ), anti-CD138 (e.g. BT062), Torcl / 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.

[0353] In some embodiments, the anticancer agent is selected from the group consisting of mechlorethamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, Sorafenib or any analog or derivative variant thereof.

[0354] 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 such as trastuzumab and pertuzumab Small molecule tyrosine kinase inhibitors, such as gefitinib Erlotinib Pilitinib, CP-654577, CP-724714, canertinib (CI 1033), HKI-272, lapatinib (GW-572016; ), PKI-166, AEE788, BMS-599626, HKI-357, BIBW2992, ARRY-334543 and JNJ-26483327.

[0355] 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.

[0356] In some embodiments, the anticancer agent is an inhibitor of a receptor tyrosine kinase (RTK) / growth factor receptor downstream member (e.g., a SHP2 inhibitor (e.g., SHP099, TNO155, RMC-4550, RMC-4630, JAB-3068, JAB-3312, RLY-1971, ERAS-601, SH3809, PF-07284892, or BBP-398, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof), SOS1 inhibitors (e.g., BI-1701963, BI-3406, SDR5, BAY-293 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.

[0357] In some embodiments, the anticancer agent is a SOS1 inhibitor. SOS1 inhibitors are known in the art. In some embodiments, the SOS1 inhibitor is selected from WO2022146698, WO2022081912, WO2022058344, WO2022026465, WO2022017519, WO2021173524, WO2021130731, WO2021127429, WO2021092115, WO2021105960, WO2021074227, WO2020180768, WO2020180770, WO2020173935, WO2020146470, WO2019201848, WO2019122129, WO 2018172250, and WO 2018115380, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug or tautomer thereof. In some embodiments, the disclosed compounds are used in combination with SOS1 inhibitors to treat K-Ras G13D cancer.

[0358] In some embodiments, the anticancer agent is an additional Ras inhibitor or a Ras vaccine, or another treatment designed to directly or indirectly reduce the carcinogenic activity of Ras. These 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 or GTP-bound state. In some embodiments, the Ras inhibitor targets Ras in an inactive or GDP-bound state. In some embodiments, the Ras inhibitor is, for example, a K-RasG12C inhibitor, such as AMG 510 (sotoracib), MRTX1257, MRTX849 (adagrasib), JNJ-74699157, LY3499446, ARS-1620, ARS-853, BPI-421286, LY3537982, JDQ443, AZ4625, JAB-21822, JAB-21000, IBI351, ERAS-3490, RMC-6291, or GDC-6036, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the Ras inhibitor is a K-Ras G12D inhibitor, such as MRTX1133 or JAB-22000, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the Ras inhibitor is a K-Ras G12V inhibitor, such as JAB-23000 or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the Ras inhibitor is RMC-6236 or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the Ras inhibitor is selected from the Ras(ON) inhibitors disclosed in the following patents incorporated herein by reference in their entirety, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug or tautomer thereof: WO2021091982, WO2021091967, WO2021091956 and WO2020132597. Other examples of Ras inhibitors are known in the art, such as disclosed in the following patents, which are incorporated herein by reference in their entirety: WO20220133038, WO2022133345, WO2022132200, WO2022119748, WO2022109485, WO2022109487, WO2022066805, WO2021190467, WO 2021185233, WO 2021180181, WO2021175199, 2021173923,WO2021169990, WO2021169963, WO2021168193, WO2021158071, WO2021155716, WO2021152149, WO2021150613, WO2021147967, WO2021147965, WO2021 143693, WO2021142252, WO2021141628, WO2021139748, WO2021139678, WO2021129824, WO2021129820, WO2021127404, WO2021126816, WO2021126799, WO2021124222, WO2021121371, WO2021121367, WO2021121330, WO2020050890, WO2020047192, WO2020035031, WO2020028706, WO2019241157, WO20192 W O2019051291, WO2018218070, WO2018217651, WO2018218071, WO2018218069, WO2018206539, WO2018143315, WO2018140600, WO2018140599, WO201814 0598, WO2018140514, WO2018140513, WO2018140512, WO2018119183, WO2018112420, WO2018068017, WO2018064510, WO2017201161, WO2017172979, WO 2017100546, WO2017087528, WO2017058807, WO2017058805, WO2017058728, WO2017058902, WO2017058792, WO2017058768, WO2017058915, WO2017015562, WO2016168540, WO2016164675, WO2016049568, WO2016049524, WO2015054572, WO2014152588, WO2014143659 and WO2013155223, or a pharmaceutically acceptable salt, solvate,Isomers (e.g., stereoisomers), prodrugs, or tautomers.

[0359] In some embodiments, therapeutic agents that can be combined with the compounds of the invention are MAP kinase (MAPK) pathway inhibitors (or "MAPK inhibitors"). These agents are known in the art. MAPK inhibitors include, but are not limited to, one or more of the MAPK inhibitors described in Cancers (Basel) 2015 Sep; 7(3): 1758-1784. For example, the MAPK inhibitor can be selected from one or more of the following: trametinib, binimetinib, selumetinib, cobimetinib, LErafAON (NeoPharm), ISIS 5132; vemurafenib, pimasertib, TAK733, RO4987655 (CH4987655); CI-1040; PD-0325901; CH5126766; MAP855; AZD6244; refametinib (RDEA 119 / BAY 86-9766); GDC-0973 / XL581; AZD8330 (ARRY-424704 / ARRY-704); RO5126766 (Roche, described in PLoS One. 2014 Nov 25; 9(11)); and GSK1120212 (or JTP-74057, described in Clin Cancer Res. 2011 Mar 1; 17(5):989-1000). The MAPK inhibitor may be PLX8394, LXH254, GDC-5573 or LY3009120.

[0360] In some embodiments, the anticancer agent is a destructive agent or a RAS-RAF-ERK or PI3K-AKT-TOR or PI3K-AKT signaling pathway inhibitor. These 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) September 2015; 7(3): 1758-1784. For example, a PI3K / AKT inhibitor may be selected from one or more of the following: NVP-BEZ235; BGT226; XL765 / SAR245409; SF1126; GDC-0980; PI-103; PF-04691502; PKI-587; GSK2126458.

[0361] In some embodiments, the anticancer agent is a PD-1 or PD-L1 antagonist. These agents are known in the art.

[0362] 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 immunotherapy. 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 can be a pan-RTK inhibitor, such as afatinib.

[0363] IGF-1R inhibitors are known in the art and include linsitinib or a pharmaceutically acceptable salt thereof.

[0364] EGFR inhibitors are known in the art and include, but are not limited to, small molecule antagonists, antibody inhibitors, or specific antisense nucleotides or siRNA. Useful antibody inhibitors of EGFR include cetuximab Panitumumab Zalutumumab, nimotuzumab and matuzumab. Other antibody-based EGFR inhibitors include any anti-EGFR antibody or antibody fragment that can partially or completely block the activation of EGFR by the natural ligand. Non-limiting examples of antibody-based EGFR inhibitors include 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 the monoclonal antibody Mab E7.6.3 (Yang, 1999, supra) or Mab C225 (ATCC Accession No. HB-8508) or an antibody or antibody fragment having binding specificity to EGFR.

[0365] Small molecule EGFR antagonists include gefitinib Erlotinib and lapatinib 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. Other 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: EP05722; EP0566226; WO96 / 33980; U.S. Pat. No. 5,747,498; WO96 / 30347; EP0787772; WO97 / 30034; WO97 / 30044; WO97 / 38994; WO97 / 49688; EP 837063; WO98 / 02434; WO97 / 38983; WO95 / 19774; WO95 / 19970; WO97 / 13771; WO98 / 02437; WO98 / 02438; WO97 / 32881; DE 19629652; WO98 / 33798; WO97 / 32880; WO97 / 32880; EP682027; WO97 / 02266; WO97 / 27199; WO98 / 07726; WO97 / 34895; WO96 / 31510; WO98 / 14449; WO98 / 14450; 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. Other non-limiting examples of small molecule EGFR inhibitors include any EGFR inhibitor described in Traxler et al., Exp. Opin. Ther. Patents 1998, 8(12): 1599-1625. In some embodiments, the EGFR inhibitor is an ERBB inhibitor. In humans, the ERBB family contains HER1 (EGFR, ERBB1), HER2 (NEU, ERBB2), HER3 (ERBB3) and HER (ERBB4).

[0366] MEK inhibitors are known in the art and include, but are not limited to, pimasetinib, selumetinib, cobimetinib trametinib Bimetinib In some embodiments, the MEK inhibitor targets a MEK mutation that is a class I MEK1 mutation selected from the group consisting of D67N, P124L, P124S, and L177V. In some embodiments, the MEK mutation is a class II MEK1 mutation selected from the group consisting of ΔE51-Q58, ΔF53-Q58, E203K, L177M, C121S, F53L, K57E, Q56P, and K57N.

[0367] PI3K inhibitors are known in the art and include, but are not limited to, wortmannin; 17-hydroxy wortmannin analogs described in WO06 / 044453; 4-[2-(1H-indazol-4-yl)-6-[[4-(methylsulfonyl)piperazin-1-yl]methyl]thieno[3,2-d]pyrimidin-4-yl]morpholine (also known as pictilisib or GDC-0941 and described in WO09 / 036082 and WO09 / 055730); 2-methyl-2-[4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5-c]quinolin-1-yl]phenyl]propionitrile (also known as BEZ 235 or NVP-BEZ235, and described in WO06 / 122806); (S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (described in WO08 / 070740); LY294002 (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (available from Axon Medchem); PI 103 hydrochloride (3-[4-(4-morpholinylpyrido[3',2':4,5]furo[3,2-d]pyrimidin-2-yl]phenol hydrochloride (purchased from Axon Medchem); PIK75 (2-methyl-5-nitro-2-[(6-bromoimidazo[1,2-a]pyridin-3-yl)methylene]-1-methylhydrazide-benzenesulfonic acid monohydrochloride) (purchased from Axon Medchem); PIK 90 (N-(7,8-dimethoxy-2,3-dihydro-imidazo[l,2-c]quinazolin-5-yl)-nicotinamide (purchased from AxonMedchem); AS-252424 (5-[l-[5-(4-fluoro-2-hydroxy-phenyl)-furan-2-yl]-methyl-(Z)-ylidene]-thiazolidine-2,4-dione (purchased from Axon Medchem); TGX-221 (7-methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido[1,2-a]pyrimidin-4-one (purchased from AxonMedchem); XL-765; and XL-147.Other PI3K inhibitors include demethoxyviridin, perifosine, CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, BKM120, XL147, XL765, Palomid529, GSK1059615, ZSTK474, PWT33597, IC87114, TGI00-115, CAL263, PI-103, GNE-477, CUDC-907, and AEZS-136.

[0368] AKT inhibitors are known in the art and include, but are not limited to, 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., WO 05 / 011700); indole-3-methanol and its derivatives (e.g., U.S. Pat. No. 6,656,963; Sarkar and Li J Nutr. 2004, 134(12 Suppl):3493S-3498S); perifosine (e.g., interferes with Akt membrane localization; 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 triciribine (TCN or API-2 or NCI identifier: NSC 154020; Yang et al., Cancer Res. 2004, 64:4394-9).

[0369] mTOR inhibitors are known in the art and include, but are not limited to, ATP-competitive mTORC1 / mTORC2 inhibitors, such as PI-103, PP242, PP30; Torin 1; FKBP12 enhancers; 4H-1-benzopyran-4-one derivatives; and rapamycin (also known as sirolimus) and its derivatives, including: temsirolimus Everolimus ( WO94 / 09010); deforolimus (also known as deforolimus or AP23573); rapamycin analogs (rapalogs), such as those disclosed in WO98 / 02441 and WO01 / 14387, such as AP23464 and AP23841; 40-(2-hydroxyethyl)rapamycin; 40-[3-hydroxy(hydroxymethyl)methylpropionate]-rapamycin (also known as CC1779); 40-epi-(tetrazolyl)-rapamycin (also known as ABT578); 32-deoxyrapamycin; 16-pentynyloxy-32(S)-dihydrorapamycin; derivatives disclosed in WO05 / 005434; U.S. Pat. Derivatives disclosed in U.S. Pat. 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 / 41807, WO96 / 41807, and WO2018204416; and phosphorus-containing rapamycin derivatives (e.g., WO05 / 016252). In some embodiments, the mTOR inhibitor is a bisteric inhibitor (see, e.g., WO2018204416, WO2019212990, and WO2019212991), such as RMC-5552, having the structure

[0370]

[0371] BRAF inhibitors that can be used in combination with the compounds of the invention are known in the art and include, for example, vemurafenib, dabrafenib, and encorafenib. BRAF may comprise a Class 3 BRAF mutation. In some embodiments, a Class 3 BRAF mutation is selected from one or more of the following amino acid substitutions in human BRAF: D287H; P367R; V459L; G466V; G466E; G466A; S467L; G469E; N581S; N581I; D594N; D594G; D594A; D594H; F595L; G596D; G596R; and A762E.

[0372] 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 key anti-apoptotic members of the B-cell lymphoma-2 (BCL-2) protein family. Overexpression of MCL-1 is closely associated with tumor progression and resistance to not only traditional chemotherapy but also targeted therapeutics including BCL-2 inhibitors such as ABT-263.

[0373] In some embodiments, the additional therapeutic agent is a SHP2 inhibitor. SHP2 inhibitors are known in the art. SHP2 is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene that contributes to a variety of cellular functions, including proliferation, differentiation, cell cycle maintenance, and migration. SHP2 has two N-terminal Src homology 2 domains (N-SH2 and C-SH2), a catalytic domain (PTP), and a C-terminal tail. The two SH2 domains control the subcellular localization and functional regulation of SHP2. The molecule exists in an inactive, self-inhibitory configuration stabilized by a binding network involving residues from the N-SH2 and PTP domains. Stimulation with, for example, cytokines or growth factors that act through receptor tyrosine kinases (RTKs) can cause exposure of the catalytic site, leading to enzyme activation of SHP2.

[0374] SHP2 participates in signal transduction through the RAS-mitogen-activated protein kinase (MAPK), i.e., JAK-STAT or phosphoinositide 3-kinase-AKT pathway. Mutations in the PTPN11 gene and subsequently in SHP2 have been identified in several human developmental diseases, such as Noonan Syndrome and Leopard Syndrome, and human cancers, such as juvenile myelomonocytic leukemia, neuroblastoma, melanoma, acute myeloid leukemia, and breast, lung, and colon cancers. Some of the mutations destabilize the autoinhibitory configuration of SHP2 and promote self-activation or enhanced growth factor-driven activation of SHP2. Therefore, SHP2 represents a particularly noteworthy target for the development of novel therapies for the treatment of various diseases, including cancer. It has been shown that the combination of SHP2 inhibitors (e.g., RMC-4550 or SHP099) and RAS pathway inhibitors (e.g., MEK inhibitors) can inhibit the proliferation of multiple cancer cell lines (e.g., pancreatic cancer, lung cancer, ovarian cancer, and breast cancer) in vitro. Therefore, combination therapy involving SHP2 inhibitors and RAS pathway inhibitors may be a general strategy for preventing tumor resistance in a variety of malignancies.

[0375] 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., Oncotarget, 2017, 8, 113734; and PCT applications: WO2022135568, WO 2021176072, WO 2021171261, WO 2021149817, WO2021148010, WO2021147879, WO 2021143823, WO 2021143701, WO2021143680, WO 2021121397, WO2021119525, WO 2021115286, WO2021110796, WO 2021088945, WO 2021073439, WO2021061706, WO2021061515, WO 2021043077, WO 2021033153、WO 2021028362、WO2021033153、WO 2021028362、WO 2021018287、WO 2020259679、WO2020249079、WO2020210384、WO 2020201991、WO 2020181283, WO2020177653, WO 2020165734, WO2020165733, WO 2020165732, WO2020156243, WO 2020156242, WO 2020108590, WO2020104635, WO2020094104, WO 2020094018, WO 2020081848, WO 2020073949, WO2020073945, WO 2020072656, WO 2020065453, WO 2020065452, WO2020063760, WO2020061103, WO 2020061101、WO 2020033828、WO2020033286、WO 2020022323、WO2019233810、WO 2019213318、WO2019183367、WO 2019183364、WO 2019182960、WO2019167000、WO2019165073、WO 2019158019、WO 2019152454、WO 2019051469、WO2019051084、WO2018218133、WO 2018172984、WO 2018160731、WO2018136265、WO2018136264、WO 2018130928、WO 2018129402、WO2018081091、WO 2018057884、WO2018013597、WO 2017216706、WO2017211303、WO 2017210134、WO 2017156397、WO2017100279、WO2017079723、WO 2017078499、WO 2016203406、WO 2016203405, WO2016203404, WO 2016196591, WO 2016191328, WO 2015107495, WO2015107494, WO2015107493, WO 2014176488, WO 2014113584, US20210085677, US10858359, US10934302, US10954243, US10988466, US11001561, US11033547, US11034705 or US11044675, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug or tautomer thereof, each of which is incorporated herein by reference.

[0376] In some embodiments, the SHP2 inhibitor binds to the active site. In some embodiments, the SHP2 inhibitor is a mixed irreversible inhibitor. In some embodiments, the SHP2 inhibitor binds to an allosteric site, such as a non-covalent allosteric inhibitor. In some embodiments, the SHP2 inhibitor is a covalent SHP2 inhibitor, such as an inhibitor targeting a cysteine ​​residue (C333) located outside the phosphatase active site. 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 is TNO155 having the following structure:

[0377] Or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is RMC-4550 having the following structure:

[0378]

[0379] Or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is RMC-4630 having the following structure:

[0380]

[0381] or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is JAB-3068 having the following structure:

[0382]

[0383] 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:

[0384]

[0385] or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is RLY-1971 having the following structure:

[0386]

[0387] Or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug or tautomer thereof. In some embodiments, the SHP2 inhibitor is ERAS-601, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug or tautomer thereof. In some embodiments, the SHP2 inhibitor is BBP-398, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug or tautomer thereof. In some embodiments, the SHP2 inhibitor is SH3809. In some embodiments, the SHP2 inhibitor is PF-07284892, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug or tautomer thereof.

[0388] In some embodiments, the additional therapeutic agent is selected from the group consisting of: MEK inhibitors, HER2 inhibitors, SHP2 inhibitors, CDK4 / 6 inhibitors, mTOR inhibitors, SOS1 inhibitors and PD-L1 inhibitors. In some embodiments, the additional therapeutic agent is selected from the group consisting of: MEK inhibitors, SHP2 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 a SOS1 inhibitor. In some embodiments, the Ras inhibitor of the present invention is used in combination with a PD-L1 inhibitor and a SOS1 inhibitor. In some embodiments, the Ras inhibitor of the present invention is used in combination with a PD-L1 inhibitor and a SHP2 inhibitor. In some embodiments, the Ras inhibitor of the present invention is used in combination with a MEK inhibitor and a SHP2 inhibitor. In some embodiments, the Ras inhibitor of the present invention is used in combination with a SHP2 inhibitor and a Ras inhibitor that inhibits multiple Ras isoforms and / or mutants (e.g., RMC-6236). In some embodiments, the cancer is colorectal cancer, and treatment comprises administering a combination of a Ras inhibitor of the present invention and a second or third therapeutic agent, such as a SHP2 inhibitor and a Ras inhibitor that inhibits multiple Ras isoforms and / or mutants. In some embodiments, the cancer is cholangiocarcinoma and treatment comprises administering a Ras inhibitor of the present invention, sorafenib, and a chemotherapeutic agent. In some embodiments, the cancer is gastric cancer and treatment comprises administering a Ras inhibitor of the present invention and a FGFR inhibitor (e.g., FGFR2i or FGFR4i). In some embodiments, the Ras inhibitor of the present invention is used in combination with immunotherapy, optionally in combination with a chemotherapeutic agent.

[0389] Proteasome inhibitors are known in the art and include, but are not limited to, carfilzomib. Bortezomib and oprozomib.

[0390] Immunotherapy includes, but is not limited to, monoclonal antibodies, immunomodulatory imides (IMiDs), GITR agonists, genetically engineered T cells (e.g., CAR-T cells), bispecific antibodies (e.g., BiTEs), and anti-PD-1 agents, anti-PD-L1 agents, anti-CTLA4 agents, anti-LAG1 agents, and anti-OX40 agents. Other immunotherapies are known in the art.

[0391] Immunomodulators (IMiDs) are a class of immunomodulatory drugs (drugs that regulate immune responses) that contain an imido group. IMiDs include thalidomide and its analogs (lenalidomide, pomalidomide, and apremilast).

[0392] Exemplary anti-PD-1 antibodies and methods of use thereof are described in Goldberg et al., Blood 2007, 110(1):186-192; Thompson et al., Clin. Cancer Res. 2007, 13(6):1757-1761; and WO06 / 121168A1), and are also described elsewhere herein.

[0393] FGFR inhibitors are known in the art, such as pemigatinib and erdafitinib, including FGFR2 inhibitors and FGFR4 inhibitors. See, for example, Cancers (Basel), June 2021; 13(12):2968.

[0394] BET inhibitors are known in the art, such as romidepsin, panobinostat and belinostat. See, for example, British J. Cancer 124: 1478 (2021).

[0395] PRMT5i inhibitors are known in the art, such as PF-0693999, PJ-68, and MRTX1719. See, for example, Biomed. Pharmacotherapy 144: 112252 (2021).

[0396] MAT2A inhibitors are known in the art, such as AG-270 and IDE397. See, for example, Exp Opin Ther Patents (2022) DOI: 10.1080 / 13543776.2022.2119127.

[0397] GITR agonists are known in the art and include, but are not limited to, GITR fusion proteins and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies), such as the GITR fusion proteins described in U.S. Pat. No. 6,111,090, U.S. Pat. No. 8,586,023, WO2010 / 003118, and WO2011 / 090754; or, for example, U.S. Pat. No. 7,025,962, EP1947183, U.S. Pat. No. 7,812,135; U.S. Pat. No. 8,388,967; U.S. Pat. No. 8,591,886; U.S. Pat. No. 7,618,632; EP1947183; 1866339 and the anti-GITR antibodies described in WO2011 / 028683, WO2013 / 039954, WO05 / 007190, WO07 / 133822, WO05 / 055808, WO99 / 40196, WO01 / 03720, WO99 / 20758, WO06 / 083289, WO05 / 115451 and WO2011 / 051726.

[0398] Another example of a therapeutic agent that can 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 regions, radionuclides, and combinations and conjugates thereof prepared by in vitro synthesis. Anti-angiogenic agents can be agonists, antagonists, allosteric modulators, toxins, or more generally can be used to inhibit or stimulate their targets (e.g., receptor or enzyme activation or inhibition), and thereby promote cell death or cell growth arrest. In some embodiments, the one or more additional therapies include anti-angiogenic agents.

[0399] Anti-angiogenic agents can be MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors and COX-II (cyclooxygenase 11) inhibitors. Non-limiting examples of anti-angiogenic agents include rapamycin, temsirolimus (CCI-779), everolimus (RAD001), sorafenib, sunitinib and bevacizumab. Examples of useful COX-II inhibitors include alecoxib, valdecoxib and rofecoxib. Examples of useful matrix metalloproteinase inhibitors are described in WO96 / 33172, WO96 / 27583, WO98 / 07697, WO98 / 03516, WO98 / 34918, WO98 / 34915, WO98 / 33768, WO98 / 30566, WO90 / 05719, WO99 / 52910, WO99 / 52889, WO99 / 29667, WO99007675, EP0606046, EP0780386, EP1786785, EP1181017, EP0818442, EP1004578 and US20090012085, and U.S. Pat. Nos. 5,863,949 and 5,861,510. Preferred MMP-2 and MMP-9 inhibitors are those with little or no MMP-1 inhibitory activity. More preferred are inhibitors that selectively inhibit MMP-2 or AMP-9 relative 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, RO 32-3555, and RS13-0830.

[0400] Other exemplary anti-angiogenic agents include kinase domain 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 (e.g., bevacizumab) or soluble VEGF receptors or their ligand binding regions), such as VEGF-TRAP TM , as well as anti-VEGF receptor agents (e.g., antibodies or antigen binding regions that specifically bind to VEGF receptors), VEGF inhibitors, EGFR inhibitors (e.g., antibodies or antigen binding regions that specifically bind to EGFR), such as (panitumumab), erlotinib Anti-Ang1 agents and anti-Ang2 agents (e.g., antibodies or antigen binding regions that specifically bind to Angl and Ang2 or their receptors, such as Tie2 / Tek), and anti-Tie2 kinase inhibitors (e.g., antibodies or antigen binding regions that specifically bind to Tie2 kinase). 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 regions that specifically bind, or soluble TWEAK receptor antagonists; see US6,727,225), ADAM disintegrin domains that antagonize the binding of integrins to their ligands (US 2002 / 0042368), specifically binding anti-eph receptor or anti-ephrin antibodies or antigen binding regions (U.S. Pat. Nos. 5,981,245, 5,728,813, 5,969,110, 6,596,852, 6,232,447, 6,057,124 and their family members), and anti-PDGF-BB antagonists (e.g., specifically binding antibodies or antigen binding regions), as well as antibodies or antigen binding regions that specifically bind to PDGF-BB ligands, and PDGFR kinase inhibitors (e.g., antibodies or antigen binding regions that specifically bind to PDGFR kinases). Additional anti-angiogenic agents include: SD-7784 (Pfizer, USA); cilengitide (Merck KGaA, Germany, EPO 0770622); pegaptanib octasodium (Gilead Sciences, USA); Alphastatin (BioActa, UK); M-PGA (Celgene, USA, US 5712291); ilomastat (Arriva, USA, US 5892112); emaxanib (Pfizer, USA, US 5792783); vatalanib (Novartis, Switzerland); 2-methoxyestradiol (EntreMed, USA); TLC ELL-12 (Elan, Ireland); anecortave acetate (Celgene, USA, US 5712291); ilomastat (Arriva, USA, US 5892112); emaxanib (Pfizer, USA, US 5792783); vatalanib (Novartis, Switzerland); 2-methoxyestradiol (EntreMed, USA); TLC ELL-12 (Elan, Ireland); anecortave acetate (Celgene, USA, US 5712291); acetate) (Alcon, USA); α-D148 Mab (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, EP 0970070); 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 (IVAX, 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); XL 647 (Exelixis, USA); second-generation α5β3 integrin MAb (Applied Molecular Evolution, USA and Medlmmune, USA); enzastaurin hydrochloride (enzastaurin hydrochloride) (Lilly, USA); CEP 7055 (Cephalon, USA and Sanofi-Synthelabo, France); BC 1 (Genoa Institute of Cancer Research, Italy); rBPI21 and BPI-derived antiangiogenic agents (XOMA, USA); PI 88 (Progen, Australia); cilengitide (Merck KGaA, Germany;Munich Technical University, Germany, Scripps Clinic and Research Foundation, USA); AVE 8062 (Ajinomoto, Japan); AS1404 (Cancer Research Laboratory, New Zealand); SG 292, (Telios, USA); endostatin (Endostatin) (Boston Childrens Hospital, USA); ATN 161 (Attenuon, USA); 2-methoxyestradiol (Boston Childrens Hospital, USA); ZD6474 (AstraZeneca, UK); ZD 6126 (Angiogene Pharmaceuticals, UK); PPI2458 (Praecis, USA); AZD 9935 (AstraZeneca, UK); AZD 2171, (AstraZeneca, UK); vatalanib (pINN) (Novartis, Switzerland and Schering AG, Germany); tissue factor pathway inhibitor (EntreMed, USA); pegaptanib (Pinn) (Gilead Sciences, USA); xanthorrhizol (Yonsei University, South Korea); gene-based VEGF-2 vaccine (Scripps Clinic and Research Foundation, USA); SPV5.2, (Supratek, Canada); SDX 103 (University of California, San Diego, USA); PX 478 (ProlX, USA); METASTATIN (EntreMed, USA); troponin I (Harvard University, USA); SU 6668 (SUGEN, USA); OXI 4503 (OXiGENE, USA); o-guanidine (Dimensional Pharmaceuticals, USA); motuporamine C (British Columbia University, Canada); CDP 791 (Celltech Group, UK); atiprimod (pINN) (GlaxoSmithKline, UK); E 7820 (Eisai, Japan);CYC 381 (Harvard University, USA); AE941 (Aeterna, Canada); angiogenesis vaccine (EntreMed, USA); urokinase plasminogen activator inhibitor (Dendreon, USA); oglufanide (pINN) (Melmotte, USA); HIF-1α inhibitor (Xenova, UK); CEP 5214 (Cephalon, USA); BAYRES2622 (Bayer, Germany); angustin (InKine, USA); A6 (Angstrom, USA); KR 31372 (Korea Research Institute of Chemical Technology, South Korea); GW2286 (GlaxoSmithKline, UK); EHT0101 (ExonHit, France); CP 868596 (Pfizer, USA); CP 564959 (OSI, USA); CP 547632 (Pfizer, USA); 786034 (GlaxoSmithKline, UK); KRN 633 (Kirin Brewery, Japan); drug delivery system, intraocular 2-methoxyestradiol; anginex (Maastricht University, Netherlands, and Minnesota University, USA); ABT510 (Abbott, USA); AAL993 (Novartis, Switzerland); VEGI (ProteomTech, USA); tumor necrosis factor-α inhibitors; SU 11248 (Pfizer, USA and SUGENUSA); ABT 518 (Abbott, USA); YH16 (Yantai Rongchang, China); S-3APG (Boston Children's Hospital, USA and EntreMed, USA); MAb, KDR (ImClone Systems, USA); MAb, α5β (Protein Design, USA); KDR kinase inhibitor (Celltech Group, UK and Johnson & Johnson, USA); GFB 116 (South Florida University, USA and Yale University, USA); CS 706 (Sankyo, Japan);combretastatin A4 prodrug (Arizona State University, USA); chondroitinase AC (IBEX, Canada); BAYRES 2690 (Bayer, Germany); AGM 1470 (Harvard University, USA, Takeda, Japan, and TAP, USA); AG 13925 (Agouron, USA); tetrathiomolybdate (University of Michigan, USA); GCS 100 (Wayne State University, USA); CV 247 (Ivy Medical, UK); CKD 732 (Chong Kun Dang, South Korea); irsogladine (Nippon Shinyaku, Japan); RG 13577 (Aventis, France); WX360 (Wilex, Germany); squalamine (Genaera, USA); RPI4610 (Sirna, USA); heparinase inhibitor (InSight, Israel); KL3106 (Kolon, South Korea); honokiol (Emory University, USA); ZKCDK (Schering AG, Germany); ZKAngio (Schering AG, Germany); ZK229561 (Novartis, Switzerland, and Schering AG, Germany); XMP 300 (XOMA, USA); VGA 1102 (Taisho, Japan); VE-cadherin-2 antagonist (ImClone Systems, USA); Vasostatin (National Institutes of Health, USA); Flk-1 (ImClone Systems, USA); TZ 93 (Tsumura, Japan); TumStatin (Beth Israel Hospital, USA); Truncated soluble FLT 1 (vascular endothelial growth factor receptor 1) (Merck & Co, USA); Tie-2 ligand (Regeneron, USA); and thrombin 1 inhibitor (Allegheny Health, Education and Research Foundation, USA). ;

[0401] Other examples of therapeutic agents that can be used in combination with the compounds of the invention include agents (e.g., antibodies, antigen binding regions, or soluble receptors) that specifically bind to and inhibit the activity of growth factors, such as antagonists of hepatocyte growth factor (HGF, also known as Scatter Factor), and antibodies or antigen binding regions that specifically bind to the receptor c-Met. These agents are known in the art.

[0402] Another example of a therapeutic agent that can be used in combination with the compounds of the invention is an autophagy inhibitor. Autophagy inhibitors are known in the art and include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil TM ), bafilomycin A1, 5-amino-4-imidazole carboxamide riboside (AICAR), soft sponge acid, autophagy inhibitory algae toxins that inhibit type 2A or type 1 protein phosphatases, cAMP analogs, and drugs that increase cAMP content, such as adenosine, LY204002, N6-mercaptopurine riboside, and vincristine. In addition, antisense RNA or siRNA that inhibits the expression of proteins, including but not limited to ATG5 (involved in autophagy), can also be used. In some embodiments, the one or more additional therapies include autophagy inhibitors.

[0403] Another example of a therapeutic agent that can be used in combination with the compounds of the invention is an anti-neoplastic agent, which are known in the art. In some embodiments, the one or more additional therapies include an anti-neoplastic agent. Non-limiting examples of antineoplastic agents include acemannan, aclarubicin, aldesleukin, alemtuzumab, alitretinoin, hexamethylmelamine, amifostine, amrubicin, amsacrine, anagrelide, anastrozole, ancer, ancestim, arglabin, arsenic trioxide, BAM-002 (Novelos), bexarotene, bicalutamide, broxuridine, capecitabine, celmoleukin, cetrorelix, cladribine, clotrimazole, cytarabine, cytos ... cytarabineocfosfate, DA3030 (Dong-A), daclizumab, denileukindiftitox, deslorelin, dexrazoxane, dilazep, docetaxel, docosanol, doxercalciferol, doxifluridine , doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HIT diclofenac, interferon alpha, daunomycin, doxorubicin, tretinoin, edelfosine, edrecolomab, eflornithine, emitefur, epirubicin, epoetin beta, etoposide phosphatephosphate), exemestane, exisulind, fadrozole, filgrastim, finasteride, fludarabine phosphate, formestane, fotemustine, gallium nitrate, gemcitabine, gemtuzumab zogamicin, gimeracil / oteracil / tegafur combination, glycopine, goserelin, heptaplatin, human chorionic gonadotropin, human fetal alpha-fetoprotein, ibandronic acidacid), idarubicin, imiquimod, interferon alpha, natural interferon alpha, interferon alpha-2, interferon alpha-2a, interferon alpha-2b, interferon alpha-Nl, interferon alpha-n3, composite interferon-1, natural interferon alpha, interferon beta, interferon beta-la, interferon beta-lb, interferon gamma, natural interferon gamma-la, interferon gamma-lb), interleukin-1 beta, iobenguane, irinotecan, irinotecan irsogladine, lanreotide, LC9018 (Yakult), leflunomide, lenograstim, lentinan sulfate, letrozole, interferon alpha, leuprorelin, levamisole + fluorouracil, liarozole, lobaplatin, lonidamine damine), lovastatin, masoprocol, melarsoprol, metoclopramide, mifepristone, miltefosine, mirimostim, mismatched double-stranded RNA, mitoguanidine, dibromodulanol, mitoxantrone, molgramostim, nafarelin, naloxone + pentazocine, nartograstim, nedaplatin, nilutamide, noscapine, novel erythropoiesis-stimulating protein, NSC631570, octreotide, oprelvekin, osaterone, oxaliplatin, paclitaxel, pamidronate acid), pegaspargase, pegylated interferon α-2b, pentosan polysulfate sodium, pentostatin, picibanil, birubicin, rabbit anti-thymocyte polyclonal antibody, pegylated interferon α-2a, porfimer sodium, raloxifene, raltitrexed, rasburiembodiment, etidronate rhenium Re186. RII: retinamide, rituximab, romurtide, samarium lexidronam, sargramostim, sizofiran, sobuzoxane, sonermin, strontium-89 chloride, suramin, tasonermin, tazarotene, tegafur, temoporfin, temozolomide, teniposide, tetrachlorodecaoxide, thalidomide, thymalfasin, thyrotropinal alpha, topotecan, toremifene, tositumomab-iodine 131), trastuzumab, treosulfan, tretinoin, trilostane, trimetrexate, triptorelin, native tumor necrosis factor alpha, ubenimex, bladder cancer vaccine, Maruyama vaccine, melanoma lysate vaccine, valrubicin, verteporfin, vinorelbine, virulizin, zinostatinstimalamer, or zoledronic acid; abarelix; AE 941 (Aeterna), ambamustine, antisense oligonucleotides, bcl-2 (Genta), APC 8015 (Dendreon), decitabine, dexaminoglutethimide, diaziquone, EL 532 (Elan), EM 800 (Endorecherche), eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant, galocitabine, gastrin 17 immunogen, HLA-B7 gene therapy (Vical), granulocyte macrophage colony stimulating factor (granulocytemacrophage colony stimulating factorcolonystimulating factor), histamine dihydrochloride, ibritumomab tiuxetan, ilomastat, IM862 (Cytran), interleukin-2, iproxifene, LDI200 (Milkhaus), leridistim, lintuzumab, CA 125MAb (Biomira), cancer MAb (Japan Pharmaceutical Development), HER-2 and Fc MAb (Medarex), idiotype 105AD7 MAb (CRC Technology), idiotype CEAM Ab (Trilex), LYM-1-iodine 131 MAb (Techniclone), polymorphic epithelial mucin-yttrium 90 MAb (Antisoma), marimastat, menogaril, mitumomab, motexafingadolinium, MX6 (Galderma), nelarabine, nolatrexed, P30 protein, pegvisomant, pemetrexed, porfiromycin, prinomastat, RL 0903 (Shire), rubitecan, satraplatin, sodium phenylacetate, spalfosic acid, SRL 172 (SR Pharma), SU5416 (SUGEN), TA077 (Tanabe), tetrathiomolybdate, thaliblastine, thrombopoietin, tinethyletiopurpurin, tirapazamine, cancer vaccine (Biomira), melanoma vaccine (NewYork University), melanoma vaccine (Sloan Kettering Institute), melanoma tumor lysate vaccine (New York Medical College), viral melanoma cell lysate vaccine (Royal Newcastle Hospital), or valspodar.

[0404] Additional examples of therapeutic agents that can be used in combination with the compounds of the invention include ipilimumab Tremelimumab; galiximab; nivolumab, also known as BMS-936558 Pembrolizumab Avelumab 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; atacicept; CP-870893; lucatumumab; dacetuzumab; muromonab-CD3; ipilumumab; MEDI4736 MSB0010718C; AMP224; adalimumab ado-trastuzumab emtansine Aflibercept Alemtuzumab Basiliximab Belimumab Basiliximab Belimumab Brentuximab vedotin Canakinumab Certolizumabpegol Daclizumab Daratumumab Denosumab Eculizumab Efalizumab Gemtuzumab ozogamicin Golimumab Ibritumomab tiuxetan infliximab Motavizumab Natalizumab Obinutuzumab Ofatumumab Omalizumab Palivizumab Pertuzumab Pertuzumab Ranibizumab Raxibacumab Tocilizumab Tositumomab; Tositumomab-i-131; Tositumomab and Tositumomab-i-131 Ustekinumab AMG 102; AMG 386; AMG 479; AMG 655; AMG 706; AMG 745; and AMG 951.

[0405] Depending on the illness being treated, the compounds described herein may be used in combination with the agents disclosed herein or other suitable agents. Therefore, in some embodiments, one or more compounds of the present invention will be co-administered with other therapies described herein. When used in combination therapy, the compounds described herein may be administered simultaneously or separately with a second agent. This combined administration may include administering two agents simultaneously in the same dosage form, administering simultaneously in independent dosage forms, and administering separately. That is, the compounds described herein and any agent described herein may be formulated into the same dosage form together and administered simultaneously. Alternatively, the compounds of the present invention and any therapy described herein may be administered simultaneously, wherein the two agents are present in independent formulations. In another alternative, the compounds of the present invention may be administered first, followed by any therapy described herein, or vice versa. In some embodiments of the separate administration scheme, the compounds of the present invention and any therapy described herein are administered a few minutes apart, a few hours apart, or a few days apart.

[0406] In some embodiments of any of the methods described herein, the first therapy (e.g., compounds of the invention) and one or more additional therapies are administered simultaneously or sequentially in any order. The first therapeutic agent may be administered immediately, at most 1 hour, at most 2 hours, at most 3 hours, at most 4 hours, at most 5 hours, at most 6 hours, at most 7 hours, at most 8 hours, at most 9 hours, at most 10 hours, at most 11 hours, at most 12 hours, at most 13 hours, 14 hours, at most 16 hours, at most 17 hours, at most 18 hours, at most 19 hours, at most 20 hours, at most 21 hours, at most 22 hours, at most 23 hours, at most 24 hours, or at most 1-7 days, 1-14 days, 1-21 days, or 1-30 days before or after administration of the one or more additional therapies.

[0407] The invention also features a kit comprising (a) a pharmaceutical composition comprising an agent described herein (e.g., a compound of the invention) and (b) a package insert with instructions for performing 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 invention), (b) one or more additional therapies (e.g., non-drug therapies or therapeutic agents) and (c) a package insert with instructions for performing any of the methods described herein.

[0408] Since one aspect of the present invention encompasses treating a disease or its associated symptoms with a combination of pharmaceutically active compounds that can be administered separately, the present invention further relates to combining independent pharmaceutical compositions in kit form. The kit may comprise two independent pharmaceutical compositions: a compound of the present invention and one or more additional therapies. The kit may comprise a container for containing the independent compositions, such as a sub-bottle or a sub-foil package. Additional examples of containers include syringes, boxes, and bags. In some embodiments, the kit may comprise instructions for the use of the independent components. The kit form is particularly advantageous when the independent components are preferably administered in different dosage forms (e.g., orally or parenterally), administered at different dosage intervals, or when the prescribing health care provider wishes to adjust the individual components in the combination.

[0409] Numbered implementation plan

[0410] 1. A compound or a pharmaceutically acceptable salt thereof having a structure of Formula I:

[0411]

[0412] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0413] X1 , X 2 and X 3 Each independently selected from CH 2 , CHF, CF 2 , C=O or O;

[0414] m is 1 or 2;

[0415] n is 0 or 1;

[0416] R 1 is hydrogen, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted 3- to 10-membered heterocycloalkyl;

[0417] R 2 is an optionally substituted C 1 -C 6 Alkyl; and

[0418] R 3 is an optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted heterocycloalkyl,

[0419] and wherein each hydrogen is independently optionally isotopically enriched with deuterium.

[0420] 2. A compound or a pharmaceutically acceptable salt thereof having a structure of Formula I:

[0421]

[0422] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0423] X 1 , X 2 and X 3 Each independently selected from CH 2 CF 2 , C=O or O;

[0424] m is 1 or 2;

[0425] n is 0 or 1;

[0426] R 1 is hydrogen, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted 3- to 10-membered heterocycloalkyl;

[0427] R 2 is an optionally substituted C 1 -C 6 Alkyl; and

[0428] R 3 is an optionally substituted C 1 -C 6 alkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted heterocycloalkyl,

[0429] and wherein each hydrogen is independently optionally isotopically enriched with deuterium.

[0430] 3. The compound or pharmaceutically acceptable salt thereof according to Embodiment 1 or 2 has a structure of any one of Formula Ia, Formula Ib or Formula Ic:

[0431]

[0432] wherein each D indicates hydrogen having an isotopic enrichment factor of at least 5 with deuterium.

[0433] 4. A compound or a pharmaceutically acceptable salt thereof as described in any one of embodiments 1 to 3, wherein R 1 is hydrogen or optionally substituted 3- to 10-membered heterocycloalkyl.

[0434] 5. A compound as described in any one of embodiments 1 to 4, or a pharmaceutically acceptable salt thereof, wherein R 1 is an optionally substituted 3- to 10-membered heterocycloalkyl group.

[0435] 6. The compound or pharmaceutically acceptable salt thereof as described in Embodiment 5, wherein R 1 yes:

[0436] 7. The compound or pharmaceutically acceptable salt thereof as described in Embodiment 5, wherein R 1 yes: and wherein each D indicates hydrogen having an isotopic enrichment factor of at least 5 with deuterium.

[0437] 8. The compound of any one of embodiments 1 to 7, or a pharmaceutically acceptable salt thereof, wherein m is 1.

[0438] 9. The compound of any one of embodiments 1 to 8, or a pharmaceutically acceptable salt thereof, wherein n is 1.

[0439] 10. A compound or a pharmaceutically acceptable salt thereof as described in any one of embodiments 1 to 9, wherein X 1 , X 2 and X 3 Each is CH2 .

[0440] 11. The compound according to Embodiment 1 or a pharmaceutically acceptable salt thereof, having a structure of Formula II:

[0441]

[0442] 12. The compound according to Embodiment 1 or a pharmaceutically acceptable salt thereof, having a structure of Formula V:

[0443]

[0444] 13. The compound or pharmaceutically acceptable salt thereof according to Embodiment 12 has a structure of any one of Formula Va, Formula Vb or Formula Vc:

[0445]

[0446]

[0447] wherein each D indicates hydrogen having an isotopic enrichment factor of at least 5 with deuterium.

[0448] 14. The compound or pharmaceutically acceptable salt thereof according to Embodiment 12 has a structure of any one of Formula Vd, Formula Ve or Formula Vf:

[0449]

[0450]

[0451] wherein each D indicates hydrogen having an isotopic enrichment factor of at least 5 with deuterium.

[0452] 15. The compound according to Embodiment 1 or a pharmaceutically acceptable salt thereof, having the structure of Formula VI:

[0453]

[0454]

[0455] 16. The compound according to Embodiment 1 or a pharmaceutically acceptable salt thereof, having the structure of Formula VII:

[0456]

[0457] 17. A compound as described in any one of embodiments 1 to 16, or a pharmaceutically acceptable salt thereof, wherein A is optionally substituted thiazole-diyl, optionally substituted oxazole-diyl, optionally substituted morpholine-diyl, optionally substituted pyrrolidine-diyl, optionally substituted piperidine-diyl or optionally substituted phenylene.

[0458] 18. The compound or pharmaceutically acceptable salt thereof as described in Embodiment 17, wherein A is optionally substituted thiazole-diyl or optionally substituted morpholine-diyl.

[0459] 19. The compound or pharmaceutically acceptable salt thereof as described in any one of embodiments 1 to 16, wherein A is an optionally substituted 5- to 10-membered heteroarylene.

[0460] 20. The compound according to embodiment 19 or a pharmaceutically acceptable salt thereof, wherein A is:

[0461] 21. The compound according to embodiment 20 or a pharmaceutically acceptable salt thereof, wherein A is:

[0462] 22. The compound of any one of embodiments 1 to 16, or a pharmaceutically acceptable salt thereof, wherein A is optionally substituted phenylene.

[0463] 23. The compound according to embodiment 22 or a pharmaceutically acceptable salt thereof, wherein A is:

[0464] 24. The compound according to embodiment 23 or a pharmaceutically acceptable salt thereof, wherein A is:

[0465] 25. The compound of any one of embodiments 1 to 16, or a pharmaceutically acceptable salt thereof, wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene.

[0466] 26. The compound of embodiment 25 or a pharmaceutically acceptable salt thereof, wherein A is an optionally substituted 6-membered heterocycloalkylene group.

[0467] 27. The compound or pharmaceutically acceptable salt thereof as described in Embodiment 25, wherein A is selected from the following or their stereoisomers:

[0468] 28. A compound or a pharmaceutically acceptable salt thereof as described in Embodiment 26, wherein A is selected from the following or their stereoisomers:

[0469] 29. A compound as described in any one of embodiments 1 to 28, or a pharmaceutically acceptable salt thereof, wherein R 2 yes:

[0470] 30. A compound as described in any one of embodiments 1 to 28, or a pharmaceutically acceptable salt thereof, wherein R2 yes: and wherein each D indicates hydrogen having an isotopic enrichment factor of at least 5 with deuterium.

[0471] 31. A compound or a pharmaceutically acceptable salt thereof as described in any one of embodiments 1 to 30, wherein R 3 is an optionally substituted C 1 -C 6 alkyl or optionally substituted 3- to 6-membered cycloalkyl.

[0472] 32. A compound as described in any one of embodiments 1 to 31, or a pharmaceutically acceptable salt thereof, wherein R 3 is an optionally substituted C 1 -C 6 alkyl.

[0473] 33. The compound or pharmaceutically acceptable salt thereof as described in embodiment 32, wherein R 3 yes:

[0474] 34. The compound or pharmaceutically acceptable salt thereof as described in embodiment 33, wherein R 3 yes:

[0475] 35. The compound or pharmaceutically acceptable salt thereof as described in embodiment 32, wherein R 3 yes: and wherein each D indicates hydrogen having an isotopic enrichment factor of at least 5 with deuterium.

[0476] 36. A compound as described in any one of embodiments 1 to 31, or a pharmaceutically acceptable salt thereof, wherein R 3 is or is an optionally substituted 3- to 6-membered cycloalkyl.

[0477] 37. The compound or pharmaceutically acceptable salt thereof as described in embodiment 36, wherein R 3 yes:

[0478] 38. The compound or pharmaceutically acceptable salt thereof as described in embodiment 36, wherein R 3 is or is an optionally substituted 5-membered cycloalkyl.

[0479] 39. The compound or pharmaceutically acceptable salt thereof as described in embodiment 38, wherein R 3 yes:

[0480] 40. The compound of any one of embodiments 11 to 16, or a pharmaceutically acceptable salt thereof, wherein:

[0481] R 2 yes

[0482] R 3 yes and

[0483] A is

[0484] 41. A compound as described in any one of embodiments 11 to 16, wherein:

[0485] R 2 yes

[0486] R 3 yes and

[0487] A is

[0488] 42. A compound as described in any one of Embodiments 1 to 41, or a pharmaceutically acceptable salt thereof, wherein the compound is not a compound of Table 3.

[0489] 43. A compound or a pharmaceutically acceptable salt thereof having the structure of a compound of Table 1 or Table 2.

[0490] 44. A pharmaceutical composition comprising a compound according to any one of embodiments 1 to 43 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0491] 45. A method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of Embodiments 1 to 43, or a pharmaceutical composition according to Embodiment 44.

[0492] 46. ​​The method of embodiment 45, wherein the cancer is pancreatic cancer, non-small cell lung cancer, colorectal cancer, or endometrial cancer.

[0493] 47. The method of embodiment 45 or 46, wherein the cancer comprises a Ras mutation.

[0494] 48. The method of embodiment 47, wherein the Ras mutation is K-Ras G12D or K-Ras G13D.

[0495] 49. A method of treating a Ras protein-related disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of Embodiments 1 to 43, or a pharmaceutical composition according to Embodiment 44.

[0496] 50. A method of inhibiting Ras protein in a cell, the method comprising contacting the cell with an effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of Embodiments 1 to 43, or a pharmaceutical composition according to Embodiment 44.

[0497] 51. The method of embodiment 49 or 50, wherein the Ras protein is K-Ras G12D or K-RasG13D.

[0498] 52. The method of embodiment 50 or 51, wherein the cell is a cancer cell.

[0499] 53. The method of embodiment 52, wherein the cancer cell is a pancreatic cancer cell, a non-small cell lung cancer cell, a colorectal cancer cell, or an endometrial cell.

[0500] 54. The method or use of any one of embodiments 45 to 53, wherein the method further comprises administering an additional anti-cancer therapy.

[0501] 55. The method of embodiment 54, wherein the additional anti-cancer therapy is an EGFR inhibitor, a second Ras inhibitor, a SHP2 inhibitor, a SOS1 inhibitor, a Raf inhibitor, a MEK inhibitor, an ERK inhibitor, a PI3K inhibitor, a PTEN inhibitor, an AKT inhibitor, an mTORC1 inhibitor, a BRAF inhibitor, a PD-L1 inhibitor, a PD-1 inhibitor, a CDK4 / 6 inhibitor, a HER2 inhibitor, or a combination thereof.

[0502] 56. The method of embodiment 54 or 55, wherein the additional anti-cancer therapy is a SHP2 inhibitor.

[0503] 57. A conjugate or a salt thereof comprising a structure of Formula III:

[0504] MP 1

[0505] Formula III

[0506] Where P 1 is a monovalent organic fraction; and

[0507] M has the structure of Formula IV:

[0508]

[0509] wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene;

[0510] X 1 , X 2 and X 3 Each independently selected from CH 2 , CHF, CF 2 , C=O or O;

[0511] m is 1 or 2;

[0512] n is 0 or 1;

[0513] R 1 is hydrogen, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted 3- to 10-membered heterocycloalkyl;

[0514] R 2 is an optionally substituted C 1 -C 6 Alkyl; and

[0515] R 3 is an optionally substituted C 1 -C 6 alkyl, optionally substituted 3 to 6 membered cycloalkyl, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted heterocycloalkyl,

[0516] and wherein each hydrogen of Formula IV is independently optionally isotopically enriched with deuterium.

[0517] 58. The conjugate or salt thereof of embodiment 57, wherein A is optionally substituted thiazole-diyl, optionally substituted oxazole-diyl, optionally substituted morpholine-diyl, optionally substituted pyrrolidine-diyl, optionally substituted piperidine-diyl, or optionally substituted phenylene.

[0518] 59. The conjugate or salt thereof as described in embodiment 57 or 58, wherein R 1 yes:

[0519]

[0520] 60. The conjugate or salt thereof of any one of embodiments 57 to 59, wherein m is 1, n is 1, and X 1 , X 2 and X 3 Each is CH 2 .

[0521] 61. The conjugate or salt thereof of any one of embodiments 57 to 60, wherein the monovalent organic moiety is a protein.

[0522] 62. The conjugate or salt thereof of embodiment 61, wherein the protein is Ras protein.

[0523] 63. The conjugate or salt thereof of embodiment 62, wherein the Ras protein is K-Ras G12D or K-RasG13D.

[0524] 64. The conjugate of any one of embodiments 57 to 63, wherein M is bound to an amino acid residue of the monovalent organic moiety.

[0525] Example

[0526] The present invention will be further illustrated by the following examples and synthetic examples, which should not be construed as limiting the scope or spirit of the present invention to the specific procedures described herein. It should be understood that the examples provided are intended to illustrate certain embodiments and are not intended to limit the scope of the present invention thereby. It should also be understood that various other embodiments, modifications thereof, and equivalents that may be conceivable to those skilled in the art may be resorted to without departing from the spirit of the present invention or the scope of the appended claims.

[0527] Chemical synthesis

[0528] Definitions used in the following examples and elsewhere herein are as follows:

[0529] CH 2 Cl 2 , DCM methylene chloride, dichloromethane

[0530] CH 3 CN, MeCN Acetonitrile

[0531] CuI Copper(I) iodide

[0532] DIPEA Diisopropylethylamine

[0533] DMF N,N-Dimethylformamide

[0534] EtOAc Ethyl acetate

[0535] h hour

[0536] H 2 O Water

[0537] HCl

[0538] K 3 PO 4 Potassium Phosphate (Tribasic)

[0539] MeOH Methanol

[0540] Na 2 SO 4 Sodium sulfate

[0541] NMP N-Methylpyrrolidone

[0542] Pd(dppf)Cl 2 [1,1'-Bis(diphenylphosphino)ferrocene]dichloropalladium(II)

[0543] Synthetic intermediates

[0544] Intermediate 1: Synthesis of 3-(5-bromo-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-3-yl)-2,2-dimethylpropan-1-ol

[0545]

[0546] Step 1: Synthesis of 1-(5-bromo-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one

[0547] In N 2 To a mixture of 3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropanoyl chloride (65 g, 137 mmol, crude) in DCM (120 mL) was slowly added 1 M SnCl 4 The mixture was stirred at 0°C for 30 minutes, then a solution of 5-bromo-1H-indole (26.8 g, 137 mmol) in DCM (40 mL) was added dropwise. The mixture was stirred at 0°C for 45 minutes, then diluted with EtOAc (300 mL), washed with brine (4×100 mL), and washed with Na 2 SO 4 The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give the product (55 g, 75% yield). LCMS (ESI): C 29 H 32 BrNO 2 m / z [M+Na] calculated for SiNa: 556.1; found: 556.3.

[0548] Step 2: Synthesis of 1-(5-bromo-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one

[0549] In N 2To a mixture of 1-(5-bromo-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one (50 g, 93.6 mmol) in THF (100 mL) was added LiBH 4 (6.1 g, 281 mmol). The mixture was heated to 60°C and stirred for 20 hours, then MeOH (10 mL) and EtOAc (100 mL) were added, and the mixture was washed with brine (50 mL), and then purified by Na 2 SO 4 The residue was diluted with DCM (50 mL), cooled to 10 °C and dihydropyridine (9.5 g, 37.4 mmol) and TsOH·H 2 O (890 mg, 4.7 mmol). The mixture was stirred at 10°C for 2 hours, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give the product (41 g, 84% yield). LCMS (ESI): C 29 H 34 m / z [M+H] calcd for BrNOSi: 519.2; found: 520.1.

[0550] Step 3: Synthesis of 5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-iodo-1H-indole

[0551] At room temperature, 1-(5-bromo-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one (1.5 g, 2.9 mmol) and 1 2 To a mixture of d-H-(4-(4-piperidin-2-yl)-1,2-d-butyl ... 2 S 2 O 3 The aqueous solution (100 mL) was washed with anhydrous Na 2 SO 4 The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give the product as a solid (900 mg, 72% yield).

[0552] Step 4: Synthesis of (1S)-1-(3-bromopyridin-2-yl)ethanol

[0553] Under Ar atmosphere, at 0 °C, HCO 2 H (66.3 g, 1.44 mol) in Et 3N (728mL, 7.2mol) was stirred in a mixture of (4S, 5S)-2-chloro-2-methyl-1-(4-methylbenzenesulfonyl)-4,5-diphenyl-1,3-diaza-2-ruthenium cyclopentane isopropyltoluene (3.9g, 6.0mmol) was added portionwise. The mixture was heated to 40°C and stirred for 15 minutes, then cooled to room temperature and 1-(3-bromopyridin-2-yl)ethanone (120g, 600mmol) was added in portions. The mixture was heated to 40°C and stirred for an additional 2 hours, then the solvent was concentrated under reduced pressure. Brine (2L) was added to the residue, the mixture was extracted with EtOAc (4×700mL), and the mixture was purified by anhydrous Na 2 SO 4 The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give the product as an oil (100 g, 74% yield). LCMS (ESI): C 7 H 8 m / z [M+H] calcd for BrNO: 201.98; found: 201.9.

[0554] Step 5: Synthesis of 3-bromo-2-[(1S)-1-methoxyethyl]pyridine

[0555] To a stirred mixture of (1S)-1-(3-bromopyridin-2-yl)ethanol (100 g, 495 mmol) in DMF (1 L) was added a 60% dispersion of NaH in oil (14.25 g, 594 mmol) in portions at 0°C. The mixture was stirred at 0°C for 1 hour. MeI (140.5 g, 990 mmol) was added dropwise at 0°C and the mixture was allowed to warm to room temperature and stirred for 2 hours. The mixture was cooled to 0°C and saturated NH 4 The mixture was extracted with EtOAc (3×1.5 L) and washed with anhydrous Na 2 SO 4 The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give the product as an oil (90 g, 75% yield). LCMS (ESI): C 8 H 10 m / z [M+H] calcd for BrNO: 215.99; found: 215.9.

[0556] Step 6: Synthesis of 2-[(1S)-1-methoxyethyl]-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine

[0557] To a stirred mixture of 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (90 g, 417 mmol) in toluene (900 mL) was added bis(pinacol)diboron (127 g, 500 mmol), KOAc (81.8 g, 833 mmol) and Pd(dppf)Cl at room temperature under Ar atmosphere. 2 (30.5 g, 41.7 mmol). The mixture was heated to 100°C and stirred for 3 hours. The filtrate was concentrated under reduced pressure and the residue was purified by Al 2 O 3 Column chromatography purification afforded the product as a semi-solid (100 g, 63% yield). LCMS (ESI): C 14 H 22 BNO 3 m / z [M+H] calcd: 264.17; found: 264.1.

[0558] Step 7: Synthesis of 5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-1H-indole

[0559] To a stirred mixture of 5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-2-iodo-1H-indole (140 g, 217 mmol) and 2-[(1S)-1-methoxyethyl]-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (100 g, 380 mmol) in dioxane (1.4 L) was added K in portions at room temperature under Ar atmosphere. 2 CO 3 (74.8g, 541mmol), Pd(dppf)Cl 2 (15.9 g, 21.7 mmol) and H 2 O (280 mL). The mixture was heated to 85 °C and stirred for 4 h, followed by the addition of cold H 2 O (5 L) and the mixture was extracted with EtOAc (3×2 L). The combined organic layers were washed with brine (2×1 L) and dried over anhydrous Na 2 SO 4 The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give the product as a solid (71 g, 45% yield). LCMS (ESI): C 37 H 43 Bn 2 O 2 m / z [M+H] calcd for Si: 655.23; found: 655.1.

[0560] Step 8: Synthesis of 5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indole

[0561] In N 2 To a stirred mixture of 5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-1H-indole (71 g, 108 mmol) in DMF (0.8 L) was added Cs in portions at 0 °C under atmosphere. 2 CO 3 (70.6 g, 217 mmol) and EtI (33.8 g, 217 mmol). The mixture was warmed to room temperature and stirred for 16 h, followed by the addition of H 2 O (4 L) and the mixture was extracted with EtOAc (3×1.5 L). The combined organic layers were washed with brine (2×1 L) and washed with anhydrous Na 2 SO 4 The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give the product as an oil (66 g, 80% yield). LCMS (ESI): C 39 H 47 Bn 2 O 2 m / z [M+H] calcd for Si: 683.26; found: 683.3.

[0562] Step 9: Synthesis of 3-(5-bromo-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-3-yl)-2,2-dimethylpropan-1-ol

[0563] In N 2 To a stirred mixture of TBAF (172.6 g, 660 mmol) in THF (660 mL) was added 5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indole (66 g, 97 mmol) in portions at room temperature under a 2% CO atmosphere. The mixture was heated to 50° C. and stirred for 16 h, cooled, and washed with H 2 O (5 L) and extracted with EtOAc (3 x 1.5 L). The combined organic layers were washed with brine (2 x 1 L) and dried over anhydrous Na 2 SO 4Dried and filtered. After filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give the product as a solid (30 g, 62% yield). LCMS (ESI) C 23 H 29 Bn 2 O 2 m / z [M+H] calcd: 445.14; found: 445.1.

[0564] Intermediate 2: Alternative synthesis via Fisher Indole Route.

[0565]

[0566] Step 1: Synthesis of 5-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-2,2-dimethyl-5-oxopentanoic acid

[0567] In N 2 To a mixture of i-PrMgCl (2M in THF, 0.5 L) was added 2.5M n-BuLi in hexanes (333 mL, 833 mmol) dropwise at -10°C over 15 minutes under a 4% CO atmosphere. The mixture was stirred at -10°C for 30 minutes, then 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (180 g, 833 mmol) in THF (0.5 L) was added dropwise at -10°C over 30 minutes. The resulting mixture was warmed to -5°C and stirred for 1 hour, then 3,3-dimethyloxane-2,6-dione (118 g, 833 mmol) in THF (1.2 L) was added dropwise at -5°C over 30 minutes. The mixture was warmed to 0°C and stirred for 1.5 hours, then quenched at 0°C by adding pre-cooled 4M HCl in dioxane (0.6 L) to adjust the pH to about 5. At 0°C, the mixture was heated with H 2 O (3 L) and extracted with EtOAc (3 x 2.5 L). The combined organic layers were washed with anhydrous Na 2 SO 4 Dry, filter, concentrate the filtrate under reduced pressure, and purify the residue by silica gel column chromatography to give the product as a solid (87 g, 34% yield). LCMS (ESI): C 15 H 21 NO 4 m / z [M+H] calcd: 280.15; found: 280.1.

[0568] Step 2: Synthesis of 3-(5-bromo-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-1H-indol-3-yl)-2,2-dimethylpropanoic acid and (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropanoic acid ethyl ester

[0569] In N 2 To a mixture of 5-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-2,2-dimethyl-5-oxopentanoic acid (78 g, 279 mmol) in EtOH (0.78 L) was added (4-bromophenyl)hydrazine hydrochloride (68.7 g, 307 mmol) in portions at room temperature under a 4% CO atmosphere. The mixture was heated to 85° C. and stirred for 2 hours, cooled to room temperature, and then a 4M HCl solution in dioxane (69.8 mL, 279 mmol) was added dropwise. The mixture was heated to 85° C. and stirred for an additional 3 hours, then concentrated under reduced pressure and the residue was dissolved in TFA (0.78 L). The mixture was heated to 60° C. and stirred for 1.5 hours, concentrated under reduced pressure and washed with saturated NaHCO 3 The residue was adjusted to about pH 5 with aqueous solution and then extracted with EtOAc (3 x 1.5 L). The combined organic layers were washed with anhydrous Na 2 SO 4 Dry, filter, concentrate the filtrate under reduced pressure, and purify the residue by silica gel column chromatography to obtain the product (78 g, crude product). LCMS (ESI) C 21 H 23 Bn 2 O 3 m / z[M+H] calculated value: 430.1 and C 23 H 27 Bn 2 O 3 m / z [M+H] calcd. for: 459.12; found: 431.1 (carboxylic acid) and 459.1.

[0570] Step 3: Synthesis of ethyl 3-(5-bromo-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-3-yl)-2,2-dimethylpropanoate

[0571] In N 2 To a mixture of 3-(5-bromo-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-1H-indol-3-yl)-2,2-dimethylpropanoic acid and (S)-ethyl 3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropanoate (198 g, 459 mmol) in DMF (1.8 L) was added Cs in portions at 0°C under atmosphere.2 CO 3 (449 g, 1.38 mol). Then, EtI (215 g, 1.38 mmol) in DMF (200 mL) was added dropwise at 0 °C. The mixture was warmed to room temperature and stirred for 4 hours, then diluted with brine (5 L) and extracted with EtOAc (3 x 2.5 L). The combined organic layers were washed with brine (2 x 1.5 L) and washed with anhydrous Na 2 SO 4 The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give the product as a solid (160 g, 57% yield). LCMS (ESI): C 25 H 31 Bn 2 O 3 m / z [M+H] calcd.: 487.17; found: 487.2.

[0572] Step 4: Synthesis of 3-(5-bromo-1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol

[0573] In N 2 To a mixture of ethyl 3-(5-bromo-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-3-yl)-2,2-dimethylpropanoate (160 g, 328 mmol) in THF (1.6 L) was added LiBH 4 (28.6 g, 1.3 mol). The mixture was heated to 60°C for 16 h, cooled, and washed with pre-cooled (0°C) saturated NH 4 The mixture was extracted with EtOAc (3×2 L), and the combined organic layers were washed with brine (2×1 L) and washed with anhydrous Na 2 SO 4 The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give two atropisomers (in the form of a single atropisomer) of 3-(5-bromo-1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol as solids (60 g, 38% yield) and (40 g, 26% yield). LCMS (ESI) m / z: C 23 H 29 Bn 2 O 2 [M+H] calcd. 445.14; found 445.2.

[0574] Intermediate 3: Synthesis (63 S,4S)-4-amino-1 1 -Ethyl-1 2 -(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzene heterocycle undecanoic acid-5,7-dione

[0575]

[0576] Step 1: Synthesis of (S)-methyl 3-(3-bromophenyl)-2-((tert-butoxycarbonyl)amino)propanoate

[0577] To a solution of (2S)-3-(3-bromophenyl)-2-[(tert-butoxycarbonyl)amino]propanoic acid (100 g, 290 mmol) in DMF (1 L) was added NaHCO at room temperature. 3 (48.8 g, 581.1 mmol) and MeI (61.9 g, 435.8 mmol). The reaction mixture was stirred for 16 h and then heated with H 2 O (1 L) and extracted with EtOAc (3×1 L). The combined organic layers were washed with brine (3×500 mL) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure. The residue was purified by silica gel column chromatography (13% EtOAc / petroleum ether) to give the final product (109 g, crude). LCMS (ESI): C 15 H 20 BrNO 4 m / z [M+Na] calcd. 380.05; found: 380.0.

[0578] Step 2: Synthesis of (S)-methyl 2-((tert-butoxycarbonyl)amino)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoate

[0579] To a stirred solution of methyl (2S)-3-(3-bromophenyl)-2-[(tert-butoxycarbonyl)amino]propanoate (108 g, 301.5 mmol) and bis(pinacol)diboron (99.53 g, 391.93 mmol) in dioxane (3.2 L) was added KOAc (73.97 g, 753.70 mmol) and Pd(dppf)Cl2 (22.06 g, 30.15 mmol). The reaction mixture was heated to 90 °C for 3 hours, then cooled to room temperature and extracted with EtOAc (2 x 3 L). The combined organic layers were washed with brine (3 x 800 mL) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure. The residue was purified by silica gel column chromatography (5% EtOAc / petroleum ether) to give the product (96 g, 78.6% yield). LCMS (ESI) C 21 H 32 BNO 6 m / z [M+Na] calcd. 428.22; found: 428.1.

[0580] Step 3: Synthesis of (S)-3-(3-(3-(3-acetoxy-2,2-dimethylpropyl)-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid methyl ester

[0581] To a solution of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]propanoate (94 g, 231.9 mmol) and 3-(5-bromo-1H-indol-3-yl)-2,2-dimethylpropyl acetate (75.19 g, 231.93 mmol) in dioxane (1.5 L) and H 2 O (300 mL) was added to the mixture 2 CO 3 (64.11 g, 463.85 mmol) and Pd(DtBPF)Cl 2 (15.12 g, 23.19 mmol). The reaction mixture was heated to 70 °C and stirred for 4 hours. The reaction mixture was extracted with EtOAc (2×2 L) and the combined organic layers were washed with brine (3×600 mL) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure. The residue was purified by silica gel column chromatography (20% EtOAc / petroleum ether) to give the product (130 g, crude). LCMS (ESI) C 30 H 38 N 2 O 6 m / z [M+H] calcd. 523.28; found: 523.1.

[0582] Step 4: Synthesis of (S)-3-(3-(3-(3-acetoxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid methyl ester

[0583] To a solution of methyl (2S)-3-(3-[3-[3-(acetyloxy)-2,2-dimethylpropyl]-1H-indol-5-yl]phenyl)-2-[(tert-butoxycarbonyl)amino]propanoate (95.0 g, 181.8 mmol) and iodine (36.91 g, 145.41 mmol) in THF (1 L) at -10 °C was added AgOTf (70.0 g, 272.7 mmol) and NaHCO 3 (22.9 g, 272.65 mmol). The reaction mixture was stirred for 30 minutes and then heated to 40 °C by adding Na 2 S 2 O 3 The resulting mixture was extracted with EtOAc (3 x 1 L) and the combined organic layers were washed with brine (3 x 500 mL) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure. The residue was purified by silica gel column chromatography (50% EtOAc / petroleum ether) to give the desired product (49.3 g, 41.8% yield). LCMS (ESI): C 30 H 37 IN 2 O 6 m / z [M+H] calcd: 649.18; found: 649.1.

[0584] Step 5: Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-3-(3-(3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl)phenyl)propanoic acid

[0585] To a solution of methyl (2S)-3-(3-[3-[3-(acetyloxy)-2,2-dimethylpropyl]-2-iodo-1H-indol-5-yl]phenyl)-2-[(tert-butoxycarbonyl)amino]propanoate (60 g, 92.5 mmol) in THF (600 mL) was added LiOH·H 2 O (19.41 g, 462.5 mmol) in H 2 O (460 mL). The resulting solution was stirred overnight and then the pH was adjusted to 6 with HCl (1 M). The resulting solution was extracted with EtOAc (2×500 mL) and the combined organic layers were washed with brine (2×500 mL) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure to obtain the product (45 g, 82.1% yield). LCMS (ESI) C 27 H 33 IN 2 O6 m / z [M+Na] calcd. 615.13; found: 615.1.

[0586] Step 6: Synthesis of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester

[0587] To a solution of (2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]phenyl]propanoic acid (30 g, 50.6 mmol) and (3S)-methyl 1,2-diazacyclohexane-3-carboxylate (10.9 g, 75.9 mmol) in DCM (400 mL) were added NMM (40.97 g, 405.08 mmol), HOBt (2.05 g, 15.19 mmol) and EDCI (19.41 g, 101.27 mmol). The reaction mixture was stirred overnight and then the mixture was washed with NH 4 Cl saturated aqueous solution (2×200 mL) and brine (2×200 mL), and the mixture was washed with Na 2 SO 4 Dry, filter and concentrate under reduced pressure to obtain the product (14 g, 38.5% yield). LCMS (ESI) C 33 H 43 IN 4 O 6 m / z [M+H] calcd. 718.23; found: 719.4.

[0588] Step 7: Synthesis of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylic acid

[0589] To a solution of (S)-methyl 1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylate (92 g, 128.0 mmol) in THF (920 mL) was added LiOH·H 2 O (26.86 g, 640.10 mmol) in H 2 O (640 mL). The reaction mixture was stirred for 2 hours and then concentrated under reduced pressure to give the product (90 g, crude). LCMS (ESI): C 32 H41 IN 4 O 6 m / z [M+H] calcd. 705.22; found: 705.1.

[0590] Step 8: Synthesis ((6 3 S,4S)-1 2 -iodo-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-tert-butyl 8-oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzheterocycloundecanoyl-4-yl)carbamate

[0591] To a solution of (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]phenyl]propanoyl]-1,2-diazacyclohexane-3-carboxylic acid (90 g, 127.73 mmol) in DCM (10 L) at 0°C was added HOBt (34.52 g, 255.46 mmol), DIPEA (330.17 g, 2554.62 mmol) and EDCI (367.29 g, 1915.96 mmol). The reaction mixture was stirred for 16 hours and then concentrated under reduced pressure. The mixture was extracted with DCM (2×2 L) and the combined organic layers were washed with brine (3×1 L) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure. The residue was purified by silica gel column chromatography (50% EtOAc / petroleum ether) to give the product (70 g, 79.8% yield). LCMS (ESI) C 32 H 39 IN 4 O 5 m / z [M+H] calcd. 687.21; found: 687.1.

[0592] Step 9: Synthesis ((6 3 S,4S)-10,10-dimethyl-5,7-dioxo-12-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-11 H-tert-butyl 8-oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzheterocycloundecanoyl-4-yl)carbamate

[0593] At room temperature, a 1L round-bottom flask was charged with ((6 3 S,4S)-1 2 -iodo-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)-indol-6(1,3)-pyridazin-2(1,3)-benzheterocycloundecyl-4-yl)carbamic acid tert-butyl ester (22.0 g, 32.042 mmol), toluene (300.0 mL), Pd 2 (dba) 3 (3.52g, 3.845mmol), S-Phos (3.95g, 9.613mmol) and KOAc (9.43g, 96.127mmol). Under stirring, 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (26.66g, 208.275mmol) was added dropwise to the mixture at room temperature. The resulting solution was stirred for 3 hours at 60°C. The resulting mixture was filtered and the filter cake was washed with EtOAc. The filtrate was concentrated under reduced pressure and the remaining residue was purified by silica gel column chromatography to give the product as a solid (22g, 90% yield). LCMS (ESI) C 38 H 51 BN 4 O 7 m / z [M+H] calcd. 687.3; found: 687.4.

[0594] Step 10: Synthesis ((6 3 S,4S)-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-tert-butyl 8-oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzheterocycloundecanoyl-4-yl)carbamate

[0595] In N 2 In the atmosphere, ((6 3 S,4S)-10,10-dimethyl-5,7-dioxo-1 2 -(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6- 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzheterocycloundecyl-4-yl)carbamic acid tert-butyl ester (2.0 g, 2.8 mmol), 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (0.60 g, 2.8 mmol), Pd(dppf)Cl 2 (0.39 g, 0.5 mmol) and K 3 PO 4 (1.2 g, 6.0 mmol) in dioxane (50 mL) and H 2 The mixture in 2% O (10 mL) was heated to 70 °C and stirred for 2 h. 2 O (50 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (3×50 mL) and dried over anhydrous Na 2 SO 4 The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give the product as a solid (1.5 g, 74% yield). LCMS (ESI): C 40 H 49 N 5 O 6 m / z [M+H] calcd. 695.4; found: 696.5.

[0596] Step 11: Synthesis ((6 3 S,4S)-1 1 -Ethyl-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-tert-butyl 8-oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzheterocycloundecanoyl-4-yl)carbamate

[0597] At 0℃, ((6 3 S,4S)-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)-indol-6(1,3)-pyridazin-2(1,3)-benzheterocycloundecyl-4-yl)carbamic acid tert-butyl ester (20 g, 28.7 mmol) and Cs 2 CO 3 To a solution of 1H-1H-2O-1 (18.7 g, 57.5 mmol) in DMF (150 mL) was added a solution of EtI (13.45 g, 86.22 mmol) in DMF (50 mL). The resulting mixture was stirred at 35 °C overnight and then treated with H 2 The mixture was extracted with EtOAc (2×300 mL) and the combined organic layers were washed with brine (3×100 mL) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure. The residue was purified by silica gel column chromatography to give the product (4.23 g, 18.8% yield) and atropisomer (5.78 g, 25.7% yield) as a solid. LCMS (ESI) C 42 H 53 N 5 O 6 m / z [M+H] calcd. 724.4; found: 724.6.

[0598] Step 12: Synthesis (6 3 S,4S)-4-amino-1 1 -Ethyl-1 2 -(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzene heterocycle undecanoic acid-5,7-dione

[0599] At 0℃, ((6 3 S,4S)-11 -Ethyl-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 A mixture of tert-butyl H-8-oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzheterocycloundecyl-4-yl)carbamate (1.3 g, 1.7 mmol) in TFA (10 mL) and DCM (20 mL) was stirred for 2 hours. The mixture was concentrated under reduced pressure to give the product as a solid (1.30 g, crude). LCMS (ESI): C 37 H 45 N 5 O 4 m / z [M+H] calcd. 623.3; found: 624.4.

[0600] Intermediate 4: Synthesis ((6 3 S,4S,Z)-1 1 -Ethyl-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-tert-butyl 8-oxa-2(4,2)-thiazol-1(5,3)-indol-6(1,3)-pyridazineheterocycleundecanoyl-4-yl)carbamate

[0601]

[0602] Step 1: Synthesis of (S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoic acid

[0603] To a solution of methyl (2S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]propanoate (110 g, 301.2 mmol) in THF (500 mL) and H 2To the solution in 200 mL of 4% dHO (200 mL) was added LiOH (21.64 g, 903.6 mmol). The resulting solution was stirred for 1 hour and then concentrated under reduced pressure. The resulting residue was adjusted to pH 6 with 1 M HCl and then extracted with DCM (3×500 mL). The combined organic layers were purified by Na 2 SO 4 Dry, filter, and concentrate under reduced pressure to give the desired product (108 g, crude). LCMS (ESI): C 11 H 15 Bn 2 O 4 m / z [M+H] calcd for S: 351.00; found: 351.0.

[0604] Step 2: Synthesis of (S)-1-((S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester

[0605] To a solution of (S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoic acid (70 g, 199.3 mmol) in DCM (500 mL) at 0°C was added (3S)-1,2-diazacyclohexane-3-carboxylic acid methyl ester bis(trifluoroacetate) salt (111.28 g, 298.96 mmol), NMM (219.12 mL. 1993.0 mmol), EDCI (76.41 g, 398.6 mmol) and HOBt (5.39 g, 39.89 mmol). The resulting solution was warmed to room temperature and stirred for 1 hour. Then H 2 The reaction was quenched with O (500 mL) and extracted with EtOAc (3×500 mL). The combined organic layers were washed with Na 2 SO 4 Dry, filter, and concentrate under reduced pressure. The residue was purified by silica gel column chromatography (0→50% EtOAc / petroleum ether) to give the desired product (88.1 g, 93% yield). LCMS (ESI): C 17 H 25 Bn 4 O 5 m / z [M+H] calcd for S: 477.08; found: 477.1.

[0606] Step 3: Synthesis of (S)-3-(1-ethyl-2-(2-(1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol

[0607] To a solution of 3-(5-bromo-1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol (60 g, 134.7 mmol) in toluene (500 mL) was added bis(pinacol)diboron (51.31 g, 202.1 mmol), Pd(dppf)Cl 2 (9.86 g, 13.48 mmol) and KOAc (26.44 g, 269.4 mmol). Then, the reaction mixture was heated to 90 ° C and stirred for 2 hours. Then, the reaction solution was cooled to room temperature and concentrated under reduced pressure. Purification by silica gel column chromatography (0→50% EtOAc / petroleum ether) gave the desired product (60.6 g, 94% yield). LCMS (ESI): C 29 H 41 BN 2 O 4 m / z [M+H] calcd: 493.32; found: 493.3.

[0608] Step 4: Synthesis of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester

[0609] To a solution of (S)-3-(1-ethyl-2-(2-(1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol (30 g, 60.9 mmol) in toluene (600 mL), dioxane (200 mL) and H2O was added at room temperature. 2 To the solution in O (200 mL) were added (S)-methyl 1-((S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (43.62 g, 91.4 mmol), K 3 PO 4 (32.23 g, 152.3 mmol) and Pd(dppf)Cl 2 (8.91 g, 12.18 mmol). The resulting solution was heated to 70 °C and stirred overnight. The reaction mixture was then cooled to room temperature and treated with H 2 O (200 mL). The resulting mixture was extracted with EtOAc (3×1000 mL), and the combined organic layers were purified by Na 2 SO 4Dry, filter, and concentrate under reduced pressure. The residue was purified by silica gel column chromatography (0→90% EtOAc / petroleum ether) to give the desired product (39.7 g, 85% yield). LCMS (ESI) m / z: C 40 H 54 N 6 O 7 [M+H] calcd. for S: 763.39; found: 763.3.

[0610] Step 5: Synthesis of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid

[0611] To a solution of (S)-methyl 1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (39.7 g, 52.0 mmol) in THF (400 mL) and H2O was added at room temperature. 2 O (100 mL) was added with LiOH·H 2 O (3.74 g, 156.2 mmol). The resulting mixture was stirred for 1.5 hours and then concentrated under reduced pressure. The residue was acidified to pH 6 with 1M HCl and extracted with DCM (3×1000 mL). The combined organic layers were washed with Na 2 SO 4 Dry, filter, and concentrate under reduced pressure to give the desired product (37.9 g, crude). LCMS (ESI): C 39 H 52 N 6 O 7 m / z [M+H] calcd for S: 749.37; found: 749.4.

[0612] Step 6: Synthesis 3 S,4S,Z)-1 1 -Ethyl-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 1H-tert-butyl 8-oxa-2(4,2)-thiazole-1(5,3)-indole-6(1,3)-pyridazineheterocyclicundecanoyl-4-yl)carbamate

[0613] To a solution of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (37.9 g, 50.6 mmol), HOBt (34.19 g, 253.0 mmol) and DIPEA (264.4 mL, 1518 mmol) in DCM (4 L) at 0°C was added EDCI (271.63 g, 1416.9 mmol). The resulting mixture was warmed to room temperature and stirred overnight. The reaction mixture was then treated with H 2 O and washed with 1M HCl (4×1 L). The organic layer was separated and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0→70% EtOAc / petroleum ether) to give the desired product (30 g, 81% yield). LCMS (ESI): C 39 H 50 N 6 O 6 m / z [M+H] calcd for S: 731.36; found: 731.3.

[0614] Intermediate 5: Synthesis (6 3 S)-4-amino-1 1 -Ethyl-1 2 -(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-2 1 ,2 2 ,2 3 ,2 6 ,6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Decahydrogen-1 1 H-8-Oxa-1(5,3)-indol-6(1,3)-pyridazin-2(5,1)-pyridine heterocycle undecanoic acid-5,7-dione

[0615]

[0616] Step 1: Synthesis of methyl 2-((tert-butoxycarbonyl)amino)acrylate

[0617] To a solution of (tert-butoxycarbonyl)-L-serine methyl ester (10 g, 45 mmol) in anhydrous MeCN (150 mL) was added DIPEA (17 g, 137 mmol). The reaction mixture was stirred at 45°C for 2 hours to give the product as a solution. LCMS (ESI): C 9 H 15 NO 4 m / z [M+Na] calcd. 201.1; found: 224.1.

[0618] Step 2: Synthesis of methyl 2-(bis(tert-butoxycarbonyl)amino)acrylate

[0619] To a solution of methyl 2-((tert-butoxycarbonyl)amino)acrylate (12 g, 60 mmol) in anhydrous MeCN (150 mL) at 0 °C was added DMAP (13 g, 90 mmol) and (Boc) 2 O (26 g, 120 mmol). The reaction was stirred for 6 hours and then H 2 O (100 mL) and extracted with DCM (3×200 mL). The combined organic layers were washed with brine (150 mL) and dried over anhydrous Na 2 SO 4 Dry, filter and concentrate under reduced pressure. The residue was purified by silica gel column chromatography to give the product as a solid (12.5 g, 65% yield). LCMS (ESI): C 14 H 23 NO 6 m / z [M+Na] calcd. 301.2; found: 324.1.

[0620] Step 3: Synthesis of methyl 2-(bis(tert-butoxycarbonyl)amino)-3-(5-bromo-3,6-dihydropyridin-1(2H)-yl)propanoate

[0621] To a mixture of 5-bromo-1,2,3,6-tetrahydropyridine (8.0 g, 49 mmol) in MeOH (120 mL) was added methyl 2-{bis[(tert-butoxy)carbonyl]amino}prop-2-enoate (22 g, 74 mmol) under Ar atmosphere. The mixture was stirred for 16 hours, then concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give the product as an oil (12 g, 47% yield). LCMS (ESI): C 19 H 31 Bn 2 O 6 m / z [M+H] calcd. 462.1; found: 463.1.

[0622] Step 4: Synthesis of 3-(5-bromo-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoic acid

[0623] To methyl 2-(bis(tert-butoxycarbonyl)amino)-3-(5-bromo-3,6-dihydropyridin-1(2H)-yl)propanoate (14 g, 30 mmol) in dioxane (30 mL) and H 2 To the mixture in 4% CO (12 mL) was added LiOH (3.6 g, 151 mmol). The mixture was heated to 35 °C and stirred for 12 hours, then 1 M HCl was added and the pH was adjusted to about 3-4. The mixture was extracted with DCM (2 x 300 mL) and the combined organic layers were washed with anhydrous Na 2 SO 4 The filtrate was concentrated under reduced pressure to give the product as a solid (10 g, 85% yield). LCMS (ESI): C 13 H 21 Bn 2 O 4 m / z [M+H] calcd. 348.1; found: 349.0.

[0624] Step 5: Synthesis of methyl (3S)-1-(3-(5-bromo-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate

[0625] To a mixture of 3-(5-bromo-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoic acid (10 g, 30 mmol), DIPEA (12 g, 93 mmol) and (3S)-1,2-diazacyclohexane-3-carboxylic acid methyl ester (5.4 g, 37 mmol) in DMF (100 mL) was added HATU (13 g, 34 mmol) at 0° C. under Ar atmosphere. The mixture was stirred at 0° C. for 2 h, followed by the addition of H 2 O and the mixture was extracted with EtOAc (2×300 mL). The combined organic layers were washed with anhydrous Na 2 SO 4 Dry, filter, concentrate the filtrate under reduced pressure and purify the residue by reverse phase chromatography to give the product as a solid (9.0 g, 55% yield). LCMS (ESI) C 19 H 31 Bn 4 O 5 m / z [M+H] calcd. 474.1; found: 475.1.

[0626] Step 6: Synthesis of (3S)-1-(2-((tert-butoxycarbonyl)amino)-3-(5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)-3,6-dihydropyridin-1(2H)-yl)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester

[0627] Under Ar atmosphere, (3S)-1-(3-(5-bromo-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester (9.0 g, 18 mmol), K 2 CO 3 (4.5 g, 32 mmol), Pd(dppf)Cl 2 DCM (1.4 g, 2 mmol), 3-(1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl)-2,2-dimethylpropan-1-ol (9.8 g, 20 mmol) in dioxane (90 mL) and H 2 The mixture in 2% O (10 mL) was heated to 75 °C and stirred for 2 h. 2 O and the mixture was extracted with EtOAc (3×200 mL). The combined organic layers were washed with Na 2 SO 4 Dry, filter, concentrate the filtrate under reduced pressure and purify the residue by silica gel column chromatography to give the product as a solid (4.0 g, 25% yield). LCMS (ESI) C 42 H 60 N 6 O 7 m / z [M+H] calcd. 760.5; found: 761.4.

[0628] Step 7: Synthesis of (3S)-1-(2-((tert-butoxycarbonyl)amino)-3-(5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)-3,6-dihydropyridin-1(2H)-yl)propanoyl)hexahydropyridazine-3-carboxylic acid

[0629] To a mixture of (3S)-methyl 1-(2-((tert-butoxycarbonyl)amino)-3-(5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)-3,6-dihydropyridin-1(2H)-yl)propanoyl)hexahydropyridazine-3-carboxylate (4.1 g, 5.0 mmol) in THF (35 mL) at 0°C was added LiOH (0.60 g, 27 mmol). The mixture was stirred at 0°C for 1.5 hours, then 1 M HCl was added to adjust the pH to about 6-7 and the mixture was extracted with EtOAc (3×200 mL). The combined organic layers were purified by Na 2 O 3 . 2 SO 4 Dry, filter and concentrate the filtrate under reduced pressure to give the product as a solid (3.6 g, 80% yield). LCMS (ESI) C 41 H 58 N 6 O 7 m / z [M+H] calcd. 746.4; found: 747.4.

[0630] Step 8: Synthesis ((6 3 S)-1 1 -Ethyl-1 2 -(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-2 1 ,2 2 ,2 3 ,2 6 ,6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Decahydrogen-1 1 H-tert-butyl 8-oxa-1(5,3)-indol-6(1,3)-pyridazin-2(5,1)-pyridine heterocycle undecyl-4-yl)carbamate

[0631] To a mixture of (3S)-1-(2-((tert-butoxycarbonyl)amino)-3-(5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)-3,6-dihydropyridin-1(2H)-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (3.6 g, 5.0 mmol) and DIPEA (24 g, 190 mmol) in DCM (700 mL) was added EDCI HCl (28 g, 140 mmol) and HOBt (6.5 g, 50 mmol) under Ar atmosphere. The mixture was heated to 30° C. and stirred at 30° C. for 16 h, then concentrated under reduced pressure. The residue was diluted with EtOAc (200 mL) and washed with H 2 O (2 × 200 mL), washed with brine (200 mL), and then washed with Na 2 SO 4 The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give the product as a solid (1.45 g, 40% yield). LCMS (ESI): C 41 H 56 N 6 O 6 m / z [M+H] calcd. 728.4; found: 729.4.

[0632] Step 9: Synthesis (6 3 S)-4-amino-1 1 -Ethyl-1 2 -(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-2 1 ,2 2 ,2 3 ,2 6 ,6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Decahydrogen-1 1 H-8-Oxa-1(5,3)-indol-6(1,3)-pyridazin-2(5,1)-pyridine heterocycle undecanoic acid-5,7-dione

[0633] At 0℃, ((6 3 S)-1 1 -Ethyl-1 2 -(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-2 1 ,2 2 ,2 3 ,26 ,6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Decahydrogen-1 1 To a mixture of tert-butyl H-8-oxa-1(5,3)-indol-6(1,3)-pyridazin-2(5,1)-pyridinyl)carbamate (130 mg, 0.20 mmol) in DCM (1.0 mL) was added TFA (0.3 mL). The mixture was warmed to room temperature and stirred for 2 hours, then concentrated under reduced pressure to give the product, which was used directly in the next step without further purification. LCMS (ESI) C 36 H 48 N 6 O 4 m / z [M+H] calcd. 628.4; found: 629.4.

[0634] Intermediate 6: Synthesis (2 2 S,6 3 S,4S)-4-amino-1 1 -Ethyl-1 2 -(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-2(4,2)-morpholino-1(5,3)-indol-6(1,3)-pyridazineheterocyclic undecanoic acid-5,7-dione

[0635]

[0636] Step 1: Synthesis of tert-butyl (2R)-2-formylmorpholin-4-ylcarboxylate

[0637] To a solution of tert-butyl (2R)-2-(hydroxymethyl)morpholin-4-ylcarboxylate (50 g, 230 mmol) in EtOAc (1 L) was added TEMPO (715 mg, 4.6 mmol) and NaHCO at room temperature. 3 (58 g, 690 mmol). The mixture was cooled to -50 °C, then TCCA (56 g, 241 mmol) in EtOAc (100 mL) was added dropwise over 30 min. The reaction mixture was warmed to 5 °C for 2 h, then treated with 10% Na 2 S 2 O3 The resulting mixture was quenched with 1% ethyl acetate (200 mL) and stirred for 20 minutes. The resulting mixture was filtered and the organic phase was separated. The aqueous phase was extracted with EtOAc (2×100 mL). The combined organic layers were washed with H 2 O (100 mL) and brine (100 mL), followed by anhydrous Na 2 SO 4 The organic layer was concentrated under reduced pressure to obtain the product (50 g, crude) as an oil.

[0638] Step 2: Synthesis of (S,Z)-2-(2-(((benzyloxy)carbonyl)amino)-3-methoxy-3-oxoprop-1-en-1-yl)morpholine-4-carboxylic acid tert-butyl ester

[0639] To a solution of tert-butyl (2R)-2-formylmorpholin-4-ylcarboxylate (49 g, 153 mmol) and methyl 2-{[(benzyloxy)carbonyl]amino}-2-(dimethoxyphosphoryl)acetate (60 g, 183 mmol) in MeCN (300 mL) was added tetramethylguanidine (35 g, 306 mmol) at 0-10°C. The reaction mixture was stirred for 30 minutes at 10°C and then warmed to room temperature for 2 hours. The reaction mixture was diluted with DCM (200 mL) and treated with 10% citric acid (200 mL) and 10% NaHCO 3 The organic phase was concentrated under reduced pressure and purified by silica gel column chromatography to give the product as a solid (36 g, 90% yield). LCMS (ESI): C 21 H 28 N 2 O 4 m / z [M+Na] Calculated value: 420.2; Found: 443.1

[0640] Step 3: Synthesis of tert-butyl (S)-2-((S)-2-(((benzyloxy)carbonyl)amino)-3-methoxy-3-oxopropyl)morpholine-4-carboxylate

[0641] To a solution of (S,Z)-2-(2-(((benzyloxy)carbonyl)amino)-3-methoxy-3-oxoprop-1-en-1-yl)morpholine-4-carboxylate (49 g, 0.12 mol) in MeOH (500 mL) was added (S,S)-Et-DUPHOS-Rh (500 mg, 0.7 mmol). 2 The mixture was stirred at room temperature under an atmosphere of (60 psi) for 48 hours. The reaction was concentrated and purified by silica gel column chromatography to give the product as a solid (44 g, 90% yield). LCMS (ESI): C 21 H 30 N2 O 7 m / z [M+Na] calcd. 422.2; found: 445.2.

[0642] Step 4: Synthesis of methyl (S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-morpholin-2-yl)propanoate

[0643] To a stirred solution of (S)-tert-butyl 2-((S)-2-(((benzyloxy)carbonyl)amino)-3-methoxy-3-oxopropyl)morpholine-4-carboxylate (2.2 g, 5.2 mmol) in EtOAc (2 mL) at 15°C was added HCl / EtOAc (25 mL). The reaction was stirred at 15°C for 2 hours and then concentrated under reduced pressure to give the product as an oil (1.51 g, 90% yield). LCMS (ESI): C 16 H 22 N 2 O 5 m / z [M+H] calcd. 322.1; found: 323.2.

[0644] Step 5: Synthesis of (S)-5-bromo-3-(3-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpropyl)-1-ethyl-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indole

[0645] To a solution of 3-(5-bromo-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-3-yl)-2,2-dimethylpropan-1-ol (100 g, 0.22 mol) and imidazole (30.6 g, 0.45 mol) in DCM (800 mL) was added TBSCl (50.7 g, 0.34 mol) in DCM (200 mL) at 0°C. The reaction was stirred at room temperature for 2 h. The resulting solution was treated with H 2 O (3 × 300 mL) and brine (2 × 200 mL), and washed with anhydrous Na 2 SO 4 Dry, filter and concentrate under reduced pressure. The residue was purified by silica gel column chromatography to give the product as a solid (138 g, 90% yield). LCMS (ESI): C 29 H 43 Bn 2 O 2 m / z [M+H] calcd for Si: 558.2; found: 559.2.

[0646] Step 6: Synthesis of methyl (2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(2S)-4-(3-{3-[(tert-butyldimethylsilyl)oxy]-2,2-dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)morpholin-2-yl]propanoate

[0647] To a stirred solution of (S)-5-bromo-3-(3-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpropyl)-1-ethyl-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indole (50 g, 89.3 mmol) in dioxane (500 mL) was added methyl (2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(2S)-morpholin-2-yl]propanoate (31.7 g, 98.2 mmol), RuPhos (16.7 g, 35.7 mmol), di-μ-chlorobis(2-amino-1,1-biphenyl-2-yl-C,N)dipalladium(II) (2.8 g, 4.4 mmol) and cesium carbonate (96 g, 295 mmol), followed by N 2 Under N atmosphere, RuPhos-Pd-G2 (3.5 g, 4.4 mmol) was added at 105°C. 2 The reaction mixture was stirred at 105° C. for 6 hours under a C atmosphere. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC chromatography to give the product as a solid (55 g, 73% yield). LCMS (ESI): C 45 H 64 N 4 O 7 m / z [M+H] calcd for Si: 800.5; found: 801.5.

[0648] Step 7: Synthesis of (2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(2S)-4-(3-{3-[(tert-butyldimethylsilyl)oxy]-2,2-dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)morpholin-2-yl]propanoic acid

[0649] To a solution of methyl (2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(2S)-4-(3-{3-[(tert-butyldimethylsilyl)oxy]-2,2-dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)morpholin-2-yl]propanoate (10 g, 12 mmol) in THF (270 mL) was added H 2The reaction was stirred at room temperature for 2 hours, and then treated with 1N HCl at 0-5°C to adjust the pH to 4-5. The resulting mixture was extracted with EtOAc (2×50 mL). The combined organic layers were washed with brine and purified by anhydrous Na 2 SO 4 Then, the organic phase was concentrated under reduced pressure to obtain the product as a solid (9.5 g, 97% yield). LCMS (ESI) C 44 H 62 N 4 O 7 m / z [M+H] calcd for Si: 786.4; found: 787.4.

[0650] Step 8: Synthesis of (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(3-(3-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpropyl)-1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester

[0651] To a stirred solution of (2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(2S)-4-(3-{3-[(tert-butyldimethylsilyl)oxy]-2,2-dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)morpholin-2-yl]propanoic acid (10 g, 12.7 mmol) in DMF (150 mL) was added (S)-methyl hexahydropyridazine-3-carboxylate (2 g, 14 mmol), followed by cooling to 0°C, adding DIPEA (32.8 g, 254 mmol), and then adding HATU (9.7 g, 25.4 mmol) at 0-5°C. The reaction mixture was stirred at 0-5°C for 1 hour. The resulting mixture was washed with EtOAc (500 mL) and H 2 O (200 mL). The organic layer was separated and 2 O (2 x 100 mL) and brine (100 mL), and dried over anhydrous sodium sulfate. The solution was filtered and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give the product. LCMS (ESI) C 50 H 72 N 6 O 8 m / z [M+H] calcd for Si: 912.5; found: 913.4.

[0652] Step 9: Synthesis of (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester

[0653] To a solution of (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(3-(3-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpropyl)-1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester (8.5 g, 9 mmol) in THF (8 mL) was added a mixture of tetrabutylammonium fluoride (1 M in THF, 180 mL, 180 mmol) and AcOH (11 g, 200 mmol) at room temperature. The reaction mixture was stirred at 75 °C for 3 h. The resulting mixture was diluted with EtOAc (150 mL) and washed with H 2 O (6 x 20 mL). The organic phase was concentrated under reduced pressure to give the product as a solid (7.4 g, 100% yield). LCMS (ESI): C 44 H 58 N 6 O 8 m / z [M+H] calcd. 799.4; found: 798.4.

[0654] Step 10: Synthesis of (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid

[0655] To a solution of (S)-methyl 1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (8 g, 10 mmol) in THF (200 mL) was added H 2The mixture was stirred at room temperature for 1 hour, then treated with 1N HCl at 0-5°C to adjust the pH to 4-5, and extracted with EtOAc (2×500 mL). The organic phase was washed with brine and concentrated under reduced pressure to give the product as a solid (8 g, crude). LCMS (ESI): C 43 H 56 N 6 O 8 m / z [M+H] calcd. 784.4; found: 785.4.

[0656] Step 11: Synthesize to obtain ((2 2 S,6 3 S,4S)-1 1 -Ethyl-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-2(4,2)-morpholino-1(5,3)-indol-6(1,3)-pyridazineheterocycleundecanoyl-4-yl)carbamic acid benzyl ester

[0657] Under argon atmosphere, to a stirred solution of (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (8 g, 10.2 mmol) and DIPEA (59 g, 459 mmol) in DCM (800 mL) at room temperature was added EDCI (88 g, 458 mmol) and HOBt (27.6 g, 204 mmol). The reaction mixture was stirred for 16 hours at room temperature. The resulting mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give the product as a solid (5 g, 66% yield); LCMS (ESI): C 43 H 54 N 6 O 7 m / z [M+H] calcd. 766.4; found: 767.4.

[0658] Step 12: Synthesis (2 2 S,6 3S,4S)-4-amino-1 1 -Ethyl-1 2 -(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-2(4,2)-morpholino-1(5,3)-indol-6(1,3)-pyridazineheterocyclic undecanoic acid-5,7-dione

[0659] In H 2 atmosphere, at room temperature to ((2 2 S,6 3 S,4S)-1 1 -Ethyl-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 To a solution of benzyl H-8-oxa-2(4,2)-morpholino-1(5,3)-indol-6(1,3)-pyridazineheterocycleundecanoyl-4-yl)carbamate (400 mg, 0.5 mmol) in MeOH (20 mL) was added Pd / C (200 mg) and ammonium acetate (834 mg, 16 mmol) and the mixture was stirred for 2 hours. The resulting mixture was filtered and concentrated under reduced pressure. The residue was redissolved in DCM (20 mL) and treated with H 2 O (5 mL x 2), and then concentrated under reduced pressure to obtain the product as a solid (320 mg, 97% yield). LCMS (ESI): C 35 H 48 N 6 O 5 m / z [M+H] calcd. 632.4; found: 633.3.

[0660] Intermediate 7: Synthesis ((6 3 S,4S,Z)-11-ethyl-1 2 -(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,64 ,6 5 ,6 6 -Hexahydro-1 1 H-tert-butyl 8-oxa-2(4,2)-thiazol-1(5,3)-indol-6(1,3)-pyridazineheterocycleundecanoyl-4-yl)carbamate

[0661]

[0662] Step 1: Synthesis of (S)-(5-bromo-6-(1-methoxyethyl)pyridin-3-yl)boronic acid

[0663] Under argon atmosphere, dtbpy (14.91 g, 55.5 mmol) and chloro(1,5-cyclooctadiene)iridium(I) dimer (7.46 g, 11.1 mmol) were added to a stirred solution of (S)-3-bromo-2-(1-methoxyethyl)pyridine (80.0 g, 370.24 mmol) and bis(pinacolato)diboron (141.03 g, 555.3 mmol) in THF (320 mL). The resulting mixture was stirred at 75 ° C for 16 hours. The mixture was concentrated under reduced pressure and the resulting residue was dissolved in EtOAc (200 mL) and washed with Na 2 CO 3 (40 g) and NaOH (10 g) in H 2 The solution in 2% O (600 mL) was adjusted to pH 10. The aqueous layer was extracted with EtOAc (800 mL), and then the aqueous phase was acidified to pH 6 with HCl (6N) to precipitate the desired product as a solid (50 g, 52% yield). LCMS (ESI): C 8 H 11 BBr 3 m / z [M+H] calcd: 260.01; found: 260.0.

[0664] Step 2: Synthesis of (S)-3-bromo-5-iodo-2-(1-methoxyethyl)pyridine

[0665] To a stirred solution of (S)-(5-bromo-6-(1-methoxyethyl)pyridin-3-yl)boronic acid (23.0 g, 88.5 mmol) in MeCN (230 mL) was added NIS (49.78 g, 221.2 mmol) at room temperature. The resulting mixture was stirred at 80 °C overnight under argon atmosphere. The mixture was concentrated under reduced pressure and the residue was dissolved in DCM (2.1 L) and treated with Na 2 S 2 O 3 The organic layer was washed with anhydrous Na 2 SO 4Dry, filter, and concentrate the filtrate under reduced pressure. The residue was purified by silica gel column chromatography to give the desired product (20 g, 66% yield). LCMS (ESI): C 8 H 9 m / z [M+H] calcd for BrINO: 341.90; found: 341.7.

[0666] Step 3: Synthesis of (S)-4-(5-bromo-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester

[0667] Into a 3L 3-necked round-bottom flask purged and maintained with an inert argon atmosphere were placed 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (147 g, 429.8 mmol), piperazine-1-carboxylic acid benzyl ester (94.69 g, 429.8 mmol), Pd(OAc) and 4-(2-(4-(2-methoxyethyl)-1-pyridine)-1-ol. 2 (4.83g, 21.4mmol), BINAP (5.35g, 8.6mmol), Cs 2 CO 3 (350.14 g, 1074.6 mmol), toluene (1 L). The resulting solution was stirred overnight at 100 ° C in an oil bath. Then, the reaction mixture was cooled to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (50% EtOAc / hexane) to give the product as a solid (135 g, 65% yield). LCMS (ESI) C 20 H 24 Bn 3 O 3 m / z [M+H] calcd. 433.1; found: 434.1.

[0668] Step 4: Synthesis of (S)-4-(6-(1-methoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester

[0669] Into a 3-L 3-necked round-bottom flask purged and maintained with an inert argon atmosphere, 4-[5-bromo-6-[(1S)-1-methoxyethyl]pyridin-3-yl]piperazine-1-carboxylic acid benzyl ester (135 g, 310.8 mmol), bis(pinacol)diboron (86.82 g, 341.9 mmol), Pd(dppf)Cl 2(22.74 g, 31.0 mmol), KOAc (76.26 g, 777.5 mmol), toluene (1 L). The resulting solution was stirred at 90 ° C in an oil bath for 2 days. The reaction mixture was cooled to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by neutral alumina column chromatography (30% EtOAC / / hexane) to give the product as a solid (167 g, crude product). LCMS (ESI) C 26 H 36 BN 3 O 5 m / z [M+H] calcd. 481.3; found: 482.1.

[0670] Step 5: Synthesis of (S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester

[0671] Into a 3-L 3-necked round-bottom flask purged and maintained with an inert argon atmosphere were placed (S)-4-(6-(1-methoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)piperazine-1-carboxylate (167 g, 346.9 mmol), 5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-2-iodo-1H-indole (224.27 g, 346.9 mmol), Pd(dppf)Cl 2 (25.38 g, 34.6 mmol), dioxane (600 mL), H 2 O (200 mL), K 3 PO 4 (184.09 g, 867.2 mmol), toluene (200 mL). The resulting solution was stirred overnight at 70 ° C in an oil bath. Then, the reaction mixture was cooled to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (50% EtOAc / hexane) to give the product as a solid (146 g, 48% yield). LCMS (ESI): C 49 H 57 Bn 4 O 4 m / z [M+H] calcd for Si: 872.3; found: 873.3.

[0672] Step 6: Synthesis of (S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1-ethyl-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester

[0673] In N 2 Under atmosphere, (S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester (146 g, 167.0 mmol) and Cs 2 CO 3 To a stirred mixture of 1,4-dihydro-1,4-dihydro-2-nitropropene (163.28 g, 501.1 mmol) in DMF (1200 mL) was added iodoethane (52.11 g, 334.0 mmol) in portions. The final reaction mixture was stirred at room temperature for 12 hours. The resulting mixture was diluted with EtOAc (1 L) and washed with brine (3×1.5 L). The organic layer was washed with anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure to give the product as a solid (143 g, crude product), which was used directly in the next step without further purification. LCMS (ESI) C 51 H 61 Bn 4 O 4 m / z [M+H] calcd for Si: 900.4; found: 901.4.

[0674] Step 7: Synthesis of (S)-4-(5-(5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester

[0675] To a stirred mixture of (S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1-ethyl-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester (143 g, 158.5 mmol) in DMF (1250 mL) was added CsF (72.24 g, 475.5 mmol). Next, in N 2The reaction mixture was stirred at 60°C for 2 days under atmosphere. The resulting mixture was diluted with EtOAc (1 L) and washed with brine (3×1 L). The organic phase was then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (30% EtOAc / petroleum ether) to give two atropisomers A (38 g, 36% yield) and B (34 g, 34% yield), both of which were solids. LCMS (ESI) C 35 H 43 Bn 4 O 4 m / z [M+H] calcd. 663.2; found: 662.2.

[0676] Step 8: Synthesis of (S)-4-(5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester

[0677] Into a 500-mL 3-necked round-bottom flask purged and maintained with an inert nitrogen atmosphere were placed (S)-4-(5-(5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester (14 g, 21.1 mmol), bis(pinacol)diboron (5.89 g, 23.21 mmol), Pd(dppf)Cl 2 (1.54 g, 2.1 mmol), KOAc (5.18 g, 52.7 mmol), toluene (150 mL). The resulting solution was stirred at 90 ° C in an oil bath for 5 hours. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (30% EtOAc / petroleum ether) to give the product as a solid (12 g, 76% yield). LCMS (ESI): C 41 H 55 BN 4 O 6 m / z [M+H] calcd. 710.4; found: 711.3.

[0678] Step 9: Synthesis of (S)-1-((S)-3-(4-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester

[0679] Into a 250-mL round-bottom flask purged and maintained with an inert argon atmosphere were placed (S)-4-(5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester (10.8 g, 15.2 mmol), (3S)-1-[(2S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]propanoyl]-1,2-diazacyclohexane-3-carboxylic acid methyl ester (7.98 g, 16.7 mmol), Pd(dtbpf)Cl 2 (0.99 g, 1.52 mmol), K 3 PO 4 (8.06 g, 37.9 mmol), toluene (60 mL), dioxane (20 mL), H 2 O (20 mL). The resulting solution was stirred at 70 ° C for 3 hours in an oil bath. The reaction mixture was cooled to room temperature. The resulting solution was extracted with EtOAc (2×50 mL) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (10% EtOAc / hexane). The solvent was removed under reduced pressure to give the product as a solid (8 g, 51% yield). LCMS (ESI): C 52 H 68 N 8 O 9 m / z [M+H] calcd. for S: 980.5; found: 980.9.

[0680] Step 10: Synthesis of (S)-1-((S)-3-(4-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid

[0681] In N 2 Under atmosphere, (S)-methyl 1-((S)-3-(4-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (12 g, 12.23 mmol) was added to a solution of THF (100 mL) / H2O 2To the stirred mixture in 0 (100 mL) was added LiOH (2.45 g, 61.1 mmol) and the resulting mixture was stirred at room temperature for 2 hours. THF was removed under reduced pressure. The pH of the aqueous phase was acidified to 5 with 1N HCl at 0°C. The aqueous layer was extracted with DCM (3×100 mL). The organic phase was concentrated under reduced pressure to give the product as a solid (10 g, 85% yield). LCMS (ESI): C 51 H 66 N 8 O 9 m / z [M+H] calcd. for S: 966.5; found: 967.0.

[0682] Step 11: Synthesis of 4-(5-((6 3 S,4S,Z)-4-((tert-butoxycarbonyl)amino)-1 1 -ethyl-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-2(4,2)-thiazolyl-1(5,3)-indolyl-6(1,3)-pyridazine heterocyclic undecylidene-1 2 -1-methoxyethyl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester

[0683] Into a 3-L round-bottom flask purged and maintained with an inert nitrogen atmosphere was placed (S)-1-((S)-3-(4-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid (18 g, 18.61 mmol), MeCN (1.8 L), DIPEA (96.21 g, 744.4 mmol), EDCI (107.03 g, 558.3 ​​mmol) and HOBt (25.15 g, 186.1 mmol). The resulting solution was stirred at room temperature overnight and then concentrated under reduced pressure. The resulting solution was diluted with DCM (1 L) and treated with 1 M HCl (3 x 1 L) and H 2 O (3×1 L). Then, the organic layer was concentrated under reduced pressure and purified by silica gel column chromatography (50% EtOAc / hexane) to obtain the product as a solid (10.4 g, 55% yield). LCMS (ESI): C 51 H64 N 8 O 8 m / z [M+H] calcd. for S: 948.5; found: 949.3.

[0684] Step 12: Synthesis ((6 3 S,4S,Z)-11-ethyl-1 2 -(2-((S)-1-methoxyethyl)-5-(piperazin-1-yl)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-tert-butyl 8-oxa-2(4,2)-thiazol-1(5,3)-indol-6(1,3)-pyridazineheterocycleundecanoyl-4-yl)carbamate

[0685] Into a 250-mL round-bottom flask purged and maintained with an inert nitrogen atmosphere was placed 4-(5-((6 3 S,4S,Z)-4-((tert-butoxycarbonyl)amino)-1 1 -ethyl-10,10-dimethyl-5,7-dioxo-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-2(4,2)-thiazolyl-1(5,3)-indolyl-6(1,3)-pyridazine heterocyclic undecylidene-1 2 -yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester (10.40 g, 10.9 mmol), Pd(OH) 2 / C (5 g, 46.9 mmol), MeOH (100 mL). 2 The resulting solution was stirred at room temperature for 3 hours under a C atmosphere. The solid was filtered off and the filter cake was washed with MeOH (3×100 mL). The combined organic phases were concentrated under reduced pressure to give the product as a solid (8.5 g, 90% yield). LCMS (ESI): C 43 H 58 N 8 O 6 m / z [M+H] calcd. for S: 814.4; found: 815.3.

[0686] Step 13: Synthesis ((63 S,4S,Z)-1 1 -Ethyl-1 2 -(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)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-tert-butyl 8-oxa-2(4,2)-thiazol-1(5,3)-indol-6(1,3)-pyridazineheterocycleundecanoyl-4-yl)carbamate

[0687] In a 1000-mL round-bottom flask purged and maintained with an inert nitrogen atmosphere, (6 3 S,4S,Z)-1 1 -Ethyl-1 2 -(2-((S)-1-methoxyethyl)-5-(piperazin-1-yl)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-tert-butyl 8-oxa-2(4,2)-thiazol-1(5,3)-indol-6(1,3)-pyridazinyl)carbamate (8.5 g, 10.4 mmol), MeOH (100 mL), AcOH (1.88 g, 31.2 mmol). The solution was stirred for 15 minutes, followed by the addition of HCHO (1.88 g, 23.15 mmol, 37% aqueous solution) and NaBH 3 CN (788 mg, 12.5 mmol). The resulting solution was stirred for 3 hours. Then, the mixture was treated with H 2 O (100 mL) and concentrated under reduced pressure to remove MeOH. The resulting solution was diluted with DCM (300 mL) and washed with H 2 O (3×100 mL). The solution was concentrated under reduced pressure to give the product as a solid (8.2 g, 90% yield). LCMS (ESI): C 44 H 60 N 8 O 6 m / z [M+H] calcd. for S: 828.4; found: 829.3.

[0688] Intermediate 8: Synthesis (63 S,4S,Z)-4-amino-1 1 -Ethyl-1 2 -(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-2(4,2)-thiazolyl-1(5,3)-indolyl-6(1,3)-pyridazineheterocyclic undecylidene-5,7-dione

[0689]

[0690] Step 1: Synthesis of (S)-3-(5-bromo-1-ethyl-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate

[0691] Under argon atmosphere, (S)-3-(5-bromo-1-ethyl-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol (100 g, 224.517 mmol) and Et 3 To a stirred solution of N (45.44 g, 449.034 mmol) in DCM (1 L) was added DMAP (2.74 g, 22.452 mmol) and Ac 2 O (27.50 g, 269.420 mmol). The resulting mixture was stirred at room temperature for 3 hours. The resulting mixture was concentrated under reduced pressure and then diluted with EtOAc (1000 mL). The resulting mixture was washed with 1M HCl (500 mL) and then with saturated NaHCO 3 (500 mL) and brine (500 mL), washed with anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by trituration with petroleum ether (500 mL) to give the product as a white solid (93.3 g, 85% yield). LCMS (ESI) C 25 H 31 Bn 2 O 3 m / z [M+H] calculated: 487.16; found: 489.2

[0692] Step 2: Synthesis of (S)-(5-(3-(3-acetoxy-2,2-dimethylpropyl)-5-bromo-1-ethyl-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)boronic acid

[0693] Under argon atmosphere, (S)-3-(5-bromo-1-ethyl-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate (93.3 g, 191.409 mmol) and B were added to the mixture at room temperature. 2 PIN 2 To a stirred solution of dtbpy (72.91 g, 287.113 mmol) in THF (370 mL) was added dtbpy (7.71 g, 28.711 mmol) and chloro(1,5-cyclooctadiene)iridium(I) dimer (6.43 g, 9.570 mmol) in portions. The resulting mixture was stirred overnight at 75 °C. The resulting mixture was concentrated under reduced pressure to give the product (190 g, crude) as an oil. LCMS (ESI): C 25 H 32 BnB 2 O 5 m / z [M+H] calcd: 531.17; found: 533.3

[0694] Step 3: Synthesis of (S)-3-(5-bromo-1-ethyl-2-(5-iodo-2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate

[0695] To a stirred solution of (S)-(5-(3-(3-acetoxy-2,2-dimethylpropyl)-5-bromo-1-ethyl-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)boronic acid (110 g, 207.059 mmol) and chloramine trihydrate-T (349.96 g, 1242.354 mmol) in THF (550 mL) at 0° C. under air atmosphere was added NaI (186.22 g, 1242.354 mmol) in H 2 O (225 mL). The resulting mixture was stirred at 50°C overnight under argon atmosphere. The resulting mixture was concentrated under reduced pressure and then washed with CHCl 3 The resulting mixture was filtered and the filter cake was washed with CHCl 3 The filtrate was washed with CHCl 3 (3×500 mL) extraction. The combined organic layers were washed with Na 2 S 2 O 3(500 mL), washed with brine (2 × 200 mL), washed with anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (18% EtOAc / petroleum ether) to give the product as a solid (24 g, 18% yield). LCMS (ESI) m / z [M+H]; C 25 H 30 BrN 2 O 3 Calculated value: 613.06; Experimental value: 614.7

[0696] Step 4: Synthesis of 3-(5-bromo-1-ethyl-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate

[0697] Under argon atmosphere, 3-(5-bromo-1-ethyl-2-{5-iodo-2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-3-yl)-2,2-dimethylpropyl acetate (9 g, 14.674 mmol), (S)-octahydropyrazino[2,1-c][1,4]oxazine (2.469 g, 17.609 mmol), Cs 2 CO 3 To a stirred solution of 1,4-dihydro- ... 2 (329.44 mg, 1.467 mmol). The resulting mixture was stirred at 100 ° C for 6 hours. After filtration, the filter cake was washed with EtOAc (100 mL). The filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (8% MeOH / DCM) to give the product as a solid (6.9 g, 75% yield). LCMS (ESI) C 32 H 43 Bn 4 O 4 m / z [M+H] calcd: 627.25; found: 627.4.

[0698] Step 5: Synthesis of 3-(1-ethyl-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate

[0699] To a stirred solution of 3-(5-bromo-1-ethyl-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate (3.2 g, 5.115 mmol), KOAc (1.51 g, 15.345 mmol) and 4,4,5,5-tetramethyl-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolan (2.60 g, 10.230 mmol) in toluene (48 mL) at room temperature was added Pd(dppf)Cl in portions under argon atmosphere. 2 (0.37 g, 0.512 mmol). The resulting mixture was stirred at 90 ° C for 1.5 hours. The resulting mixture was filtered and the filter cake was washed with EtOAc (100 mL). The filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (8% MeOH / DCM) to give the product as a solid (3.0 g, 88% yield). LCMS (ESI): C 38 H 55 BN 4 O 6 m / z [M+H] calculated: 675.43; found: 675.1

[0700] Step 6: Synthesis of (S)-1-((S)-3-(4-(3-(3-acetoxy-2,2-dimethylpropyl)-1-ethyl-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester

[0701] Under argon atmosphere, 3-(1-ethyl-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate (5 g, 7.433 mmol) was added to the mixture at room temperature. 3 PO 4 To a stirred mixture of 2-nitropropene (4.26 g, 20.067 mmol) in toluene (54 mL) were added dioxane (17.82 mL, 210.307 mmol) and H 2O (17.82 mL). The resulting mixture was stirred at 70 ° C for 2 hours. The resulting mixture was filtered and the filter cake was washed with EtOAc (100 mL). The filtrate was concentrated under reduced pressure. The resulting mixture was extracted with EtOAc (200 mL). The combined organic layer was washed with brine (100 mL) and dried over anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (8% MeOH / DCM) to give the product as a solid (4.6 g, 66% yield). LCMS (ESI): C 49 H 68 N 8 O 9 m / z [M+H] calculated for S: 945.49; found: 945.7

[0702] Step 7: Synthesis of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid

[0703] To a stirred solution of (S)-methyl 1-((S)-3-(4-(3-(3-acetoxy-2,2-dimethylpropyl)-1-ethyl-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (6 g, 6.361 mmol) in THF (43 mL) was added LiOH·H 2 O (573.92 mg, 13.677 mmol). The resulting mixture was stirred at room temperature for 16 hours. The mixture was acidified to pH 6 with HCl (aq). The resulting mixture was extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine and purified by anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure to obtain a solid product (4 g, crude product). LCMS (ESI) C 45 H 60 N 8 O 9 m / z [M+H] calculated for S: 889.43; found: 889.7

[0704] Step 8: Synthesis ((6 3 S,4S,Z)-1 1 -Ethyl-1 2 -(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-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-tert-butyl 8-oxa-2(4,2)-thiazol-1(5,3)-indol-6(1,3)-pyridazineheterocycleundecanoyl-4-yl)carbamate

[0705] To a stirred solution of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (4 g, 4.51 mmol), HOBt (6.09 g, 45.09 mmol) and DIPEA (23.31 g, 180.36 mmol) in DCM (200 mL) was added dropwise EDCI (25.93 g, 135.27 mmol) in DCM (200 mL) under argon atmosphere at 0 °C. The resulting mixture was stirred at room temperature for 16 hours and then concentrated under reduced pressure. 2 The reaction was quenched with O and extracted with EtOAc (500 mL). The combined organic layers were washed with brine and purified by anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (8% MeOH / DCM) to give the product as a solid (2.0 g, 52% yield). LCMS (ESI): C 46 H 62 N 8 O 7 m / z [M+H] calculated for S: 870.4; found: 871.8

[0706] Step 9: Synthesis (6 3 S,4S,Z)-4-amino-1 1 -Ethyl-1 2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-2(4,2)-thiazolyl-1(5,3)-indolyl-6(1,3)-pyridazineheterocyclic undecylidene-5,7-dione

[0707] Under argon atmosphere, at 0 °C, 3 S,4S,Z)-1 1 -Ethyl-1 2 -(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-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 To a stirred solution of tert-butyl H-8-oxa-2(4,2)-thiazol-1(5,3)-indol-6(1,3)-pyridazinyl)carbamate (316 mg, 0.345 mmol) in DCM (3 mL) was added TFA (1 mL) dropwise. The resulting mixture was stirred at room temperature for 2 hours. The mixture was washed with NaHCO 3 The saturated aqueous solution was basified to pH 8. The resulting mixture was extracted with EtOAc (3×80 mL). The combined organic layers were washed with brine (3×40 mL) and purified by anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The crude product mixture was used directly in the next step without further purification. LCMS (ESI) C 41 H 54 N 8 O 5 m / z [M+H] calculated for S: 771.4; found: 771.6

[0708] Intermediate 9: Synthesis (6 3 S,4S,Z)-4-amino-1 1 -Ethyl-1 2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-10,10-dimethyl-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-2(4,2)-thiazolyl-1(5,3)-indolyl-6(1,3)-pyridazineheterocyclic undecylidene-5,7-dione

[0709]

[0710] Step 1: Synthesis of 3-(5-bromo-1-ethyl-2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate

[0711] Under argon atmosphere, 3-(5-bromo-1-ethyl-2-{5-iodo-2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-3-yl)-2,2-dimethylpropyl acetate (9 g, 14.674 mmol), (R)-octahydro-2H-pyrido[1,2-a]pyrazine (2.469 g, 17.609 mmol), Cs 2 CO 3 To a stirred solution of 4-(11.9523 g, 36.685 mmol) and BINAP (456.85 mg, 0.734 mmol) in toluene (63 mL) was added Pd(OAc)2 (329.44 mg, 1.467 mmol) in portions. The resulting mixture was stirred at 100 °C for 6 hours, then the mixture was filtered and the filter cake was washed with EtOAc (100 mL). The filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (8% MeOH / DCM) to give the product as a solid (6 g, 65% yield). LCMS (ESI): C 33 H 45 Bn 4 O 3 m / z [M+H] calcd: 625.28; found: 627.4.

[0712] Step 2: Synthesis of 3-(1-ethyl-2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate

[0713] To a stirred solution of 3-(5-bromo-1-ethyl-2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate (3.2 g, 5.115 mmol), KOAc (1.51 g, 15.345 mmol) and 4,4,5,5-tetramethyl-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolan (2.60 g, 10.230 mmol) in toluene (48 mL) at room temperature was added Pd(dppf)Cl in portions under argon atmosphere. 2 (0.37 g, 0.512 mmol). The resulting mixture was stirred at 90 ° C for 1.5 hours. The resulting mixture was filtered and the filter cake was washed with EtOAc (100 mL). The filtrate was concentrated under reduced pressure and purified by preparative TLC (8% MeOH / DCM) to give the product as a solid (3.1 g, 81% yield). LCMS (ESI): C 39 H 57 BN 4 O 5 m / z [M+H] calculated: 673.45; found: 673.4

[0714] Step 3: Synthesis of (S)-1-((S)-3-(4-(3-(3-acetoxy-2,2-dimethylpropyl)-1-ethyl-2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester

[0715] Under argon atmosphere, 3-(1-ethyl-2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate (5 g, 7.433 mmol), (S)-methyl 1-((S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (3.89 g, 8.176 mmol) and K were added to the mixture at room temperature. 3 PO 4 (4.26 g, 20.067 mmol) in toluene (54 mL), dioxane (18 mL) and H 2 To the stirred mixture in 2% O (18 mL) was added Pd(dtbpf)Cl 2 (969 mg, 1.486 mmol). The resulting mixture was stirred at 70 °C for 2 hours. The mixture was filtered and the filter cake was washed with EtOAc (100 mL). The filtrate was concentrated under reduced pressure and the resulting mixture was extracted with EtOAc (200 mL). The combined organic layers were washed with brine (100 mL) and purified by anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (8% MeOH / DCM) to give the product as a solid (6.8 g, 83% yield). LCMS (ESI): C 50 H 70 N 8 O 8 m / z [M+H] calculated for S: 943.51; found: 943.4

[0716] Step 4: Synthesis of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid

[0717] To a stirred solution of (S)-methyl 1-((S)-3-(4-(3-(3-acetoxy-2,2-dimethylpropyl)-1-ethyl-2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (6 g, 6.361 mmol) in THF (43 mL) was added LiOH·H dropwise at 0° C. under argon atmosphere. 2 O (573.92 mg, 13.677 mmol). The resulting mixture was stirred at room temperature for 16 hours. The mixture was acidified to pH 6 with HCl (aq). The resulting mixture was extracted with EtOAc (3×150 mL). The combined organic layers were washed with brine and purified by anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure to obtain a solid product (4 g, crude product). LCMS (ESI) C 47 H 66 N 8 O 7 m / z [M+H] calculated for S: 887.49; found: 887.6

[0718] Step 5: Synthesis ((6 3 S,4S,Z)-1 1 -Ethyl-1 2 -(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)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-tert-butyl 8-oxa-2(4,2)-thiazol-1(5,3)-indol-6(1,3)-pyridazineheterocycleundecanoyl-4-yl)carbamate

[0719] To a stirred solution of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (4 g, 4.509 mmol), HOBt (6.09 g, 45.090 mmol) and DIPEA (23.31 g, 180.360 mmol) in DCM (200 mL) was added dropwise EDCI (25.93 g, 135.270 mmol) in DCM (200 mL) under argon atmosphere at 0 °C. The resulting mixture was stirred at room temperature for 16 hours. The resulting mixture was concentrated under reduced pressure and washed with H 2 O quenched and extracted with EtOAc (500 mL). The combined organic layers were washed with brine and 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (8% MeOH / DCM) to give the product as a solid (2.0 g, 49% yield). LCMS (ESI): C 47 H 64 N 8 O 6 m / z [M+H] calculated for S: 869.47; found: 869.8

[0720] Step 6: Synthesis (6 3 S,4S,Z)-4-amino-1 1 -Ethyl-1 2 -(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-10,10-dimethyl-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-2(4,2)-thiazolyl-1(5,3)-indolyl-6(1,3)-pyridazineheterocyclic undecylidene-5,7-dione

[0721] At 0℃, ((6 3 S,4S,Z)-1 1 -Ethyl-1 2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-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 stirred solution of tert-butyl H-8-oxa-2(4,2)-thiazol-1(5,3)-indol-6(1,3)-pyridazinyl)carbamate (900 mg, 1.035 mmol) in DCM (9 mL) was added TFA (3 mL) dropwise. The resulting mixture was stirred for 2 h at room temperature. The mixture was washed with NaHCO 3 The saturated aqueous solution was basified to pH = 8 and extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine (3×50 mL) and dried over anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure to give the product (800 mg), which was used directly in the next step without further purification. LCMS (ESI) C 42 H 56 N 8 O 4 m / z [M+H] calculated for S: 769.42; found: 769.5

[0722] Intermediate 10: Synthesis (6 3 S,4S)-4-amino-1 2 -(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-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)-indol-6(1,3)-pyridazin-2(1,3)-benzene heterocycle undecanoic acid-5,7-dione

[0723]

[0724] Step 1: Synthesis of (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate

[0725] Under argon atmosphere, (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol (60 g, 0.12 mol) and Et 3 DMAP (1.46 g, 0.012 mol) and acetic anhydride (14.7 g, 144 mmol) were added dropwise to a stirred solution of N (24.33 g, 0.24 mol) in DCM (600 mL). The resulting mixture was stirred for 2 hours at room temperature. The resulting mixture was concentrated under reduced pressure and washed with HCl (500 mL). The resulting mixture was washed with NaHCO 3 The combined organic layers were washed with brine (500 mL) and washed with anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure to obtain the product as an oil (59.6 g, 92% yield). LCMS (ESI) C 25 H 28 F 3 N 2 O 3 m / z [M+H] calculated: 541.13; found: 543.2

[0726] Step 2: Synthesis of (S)-(5-(3-(3-acetoxy-2,2-dimethylpropyl)-5-bromo-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)boronic acid

[0727] To a stirred mixture of (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate (55.1 g, 101.771 mmol) and 4,4,5,5-tetramethyl-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolan (38.77 g, 152.656 mmol) in THF (40 mL) was added dtbpy (4.10 g, 15.266 mmol) and chloro(1,5-cyclooctadiene)iridium(I) dimer (3.42 g, 5.089 mmol) in portions at room temperature under argon atmosphere. The resulting mixture was stirred at 75° C. for 5 hours. The resulting mixture was concentrated under reduced pressure to give the product (102.4 g, crude) as an oil. LCMS (ESI): C 25 H 29 BF 3 N 2 O 5 m / z [M+H] calculated: 585.14; found: 585.2

[0728] Step 3: Synthesis of (S)-3-(5-bromo-2-(5-iodo-2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate

[0729] To a stirred solution of (S)-(5-(3-(3-acetoxy-2,2-dimethylpropyl)-5-bromo-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)boronic acid (51.2 g, 87.487 mmol) and sodium chloro[(4-methylbenzene)sulfonyl]imide (197 g, 699.896 mmol) in THF (258 mL) was added dropwise in H2O at 0 °C under argon atmosphere. 2 The resulting mixture was stirred at 55°C for 16 hours. The resulting mixture was concentrated under reduced pressure and washed with CH 3 The combined organic layers were washed with brine (2×20 mL) and purified by anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (20% EtOAc / petroleum ether) to give the product as a solid (15.3 g, 26% yield). LCMS (ESI): C 32 H 40 F3 N 4 O 4 m / z [M+H] calculated: 666.0; found: 667.3

[0730] Step 4: Synthesis of (S)-1-((S)-3-(3-(3-(3-acetoxy-2,2-dimethylpropyl)-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester

[0731] To a stirred mixture of (S)-3-(5-bromo-2-(5-iodo-2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate (2.70 g, 4.046 mmol) and (S)-octahydropyrazino[2,1-c][1,4]oxazine dihydrochloride (1.044 g, 4.855 mmol) in toluene (18.9 mL) was added Cs(II) in portions at room temperature under argon atmosphere. 2 CO 3 (5932.38 mg, 18.207 mmol) and BINAP (125.97 mg, 0.202 mmol). Pd(OAc) was added to the above mixture in several portions. 2 (90.84 mg, 0.405 mmol). The resulting mixture was stirred at 90 ° C for another 16 hours. The mixture was cooled to room temperature and then filtered, and the filter cake was washed with EtOAc (2×20 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (10% MeOH / DCM) to give the product as a solid (2.3 g, 83% yield). LCMS (ESI): C 32 H 40 F 3 N 4 O 4 m / z [M+H] calculated: 681.23; found: 681.4

[0732] Step 5: Synthesis of (S)-1-((S)-3-(3-(3-(3-acetoxy-2,2-dimethylpropyl)-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester

[0733] In a 250 mL 3-necked round-bottom flask under air atmosphere, (S)-methyl 1-((S)-3-(3-(3-(3-acetoxy-2,2-dimethylpropyl)-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (2.33 g, 4.512 mmol) and K were added at room temperature. 3 PO 4 (1.59 g, 7.490 mmol). At room temperature, H 2 To a stirred mixture of 2H2O (8.20 mL) and dioxane (8.20 mL) in toluene was added Pd(dtbpf)Cl in portions. 2 (0.29 g, 0.451 mmol). The resulting mixture was stirred at 65°C for 3 hours. The resulting mixture was filtered and the filter cake was washed with EtOAc (2×100 mL). The filtrate was concentrated under reduced pressure. The resulting mixture was extracted with EtOAc (2×100 mL). The combined organic layers were washed with brine (2×150 mL) and purified by anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (3→4% MeOH / DCM) to give the product as a solid (2.7 g, 90% yield). LCMS (ESI) C 52 H 68 F 3 N 7 O 9 m / z [M+H] calculated: 991.5; found: 992.7

[0734] Step 6: Synthesis of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylic acid

[0735] Into a 100 mL 3-necked round-bottom flask were added (S)-methyl 1-((S)-3-(3-(3-(3-acetoxy-2,2-dimethylpropyl)-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (3 g, 3.024 mmol) and THF (30 mL) at room temperature. Subsequently, H 2 O (30 mL) was added in portions at 0°C. 2 The resulting mixture was stirred at room temperature for 16 hours. The mixture was acidified to pH 5 with 1N HCl. The resulting mixture was extracted with EtOAc (2×100 mL). The combined organic layers were washed with brine (2×100 mL) and purified by anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure to obtain the product as a solid (2.7 g, crude product). LCMS (ESI) C 49 H 64 F 3 N 7 O 8 m / z [M+H] calcd: 936.48; found: 936.7

[0736] Step 7: Synthesis ((6 3 S,4S)-12-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-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 1H-tert-butyl 8-oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzheterocycloundecanoyl-4-yl)carbamate

[0737] To a 2L 3-necked round-bottom flask was added (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylic acid (3.12 g, 3.333 mmol) and DCM (624 mL) at room temperature. To the above mixture were added DIPEA (17.23 g, 133.320 mmol) and HOBt (4.50 g, 33.330 mmol) in portions at 0°C. The resulting mixture was stirred for another 30 minutes. To the above mixture was added EDCI (19.17 g, 99.990 mmol) in portions at room temperature over 16 hours. The resulting mixture was concentrated under reduced pressure. 2 The reaction was quenched with 4% CO. The resulting mixture was extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine (3×100 mL) and purified by anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (3→4% MeOH / DCM) to give the product as a solid (3 g, 98% yield). LCMS (ESI): C 49 H 62 F 3 N 7 O 7 m / z [M+H] calcd: 918.47; found: 918.8

[0738] Step 8: Synthesis (6 3 S,4S)-4-amino-1 2 -(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-1 1 -(2,2,2-trifluoroethyl)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1H-8-Oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzene heterocycle undecanoic acid-5,7-dione

[0739] Under argon atmosphere, at 0 °C, 3 S,4S)-12-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-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 stirred solution of tert-butyl H-8-oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzheterocycloundecyl-4-yl)carbamate (930 mg, 1.013 mmol) in DCM (15 mL) was added TFA (5 mL, 67.315 mmol) dissolved in DCM (5 mL) dropwise. The resulting mixture was stirred at 0° C. for 2 h. The residue was washed with NaHCO 3 The saturated aqueous solution was basified to pH 8. The resulting mixture was extracted with DCM, and the combined organic layers were washed with brine and purified by anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure to obtain the product as a solid (880 mg, crude product). LCMS (ESI): C 44 H 54 F 3 N 7 O 5 m / z [M+H] calculated: 818.42; found: 818.6

[0740] Intermediate 11: Synthesis (6 3 S,4S)-4-amino-12-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-10,10-dimethyl-1 1 -(2,2,2-trifluoroethyl)-6 1 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1H-8-Oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzene heterocycle undecanoic acid-5,7-dione

[0741]

[0742] Step 1: Synthesis of (S)-(5-(3-(3-acetoxy-2,2-dimethylpropyl)-5-bromo-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)boronic acid

[0743] To a 100 mL 3-necked round bottom flask was added 3-(5-bromo-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}-1-(2,2,2-trifluoroethyl)indol-3-yl)-2,2-dimethylpropyl acetate (10 g, 18.470 mmol), 4,4,5,5-tetramethyl-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolan (8.44 g, 33.25 mmol) and dtbpy (0.89 g, 3.325 mmol) at room temperature. Chloro(1,5-cyclooctadiene)iridium(I) dimer (0.74 g, 1.108 mmol) and THF (40 mL) were added to the above mixture. The resulting mixture was stirred for another 16 hours at 80°C. The resulting mixture was concentrated under reduced pressure. The crude product was used directly in the next step without further purification. LCMS (ESI) C 25 H 29 BF 3 N 2 O 5 m / z [M+H] calcd: 585.14; found: 585.0

[0744] Step 2: Synthesis of (S)-3-(5-bromo-2-(5-iodo-2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate

[0745] To a stirred solution of (S)-(5-(3-(3-acetoxy-2,2-dimethylpropyl)-5-bromo-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)boronic acid (17.9 g, 30.586 mmol) in THF (89.5 mL) was added dropwise HCl at 0 °C under nitrogen atmosphere. 2Sodium chloro[(4-methylbenzene)sulfonyl]imide (68.93 g, 244.688 mmol) and NaI (36.68 g, 244.688 mmol) in 4% HCl (44.75 mL). The resulting mixture was stirred for another 20 minutes at room temperature and then heated to 50° C. for 16 hours. The resulting mixture was concentrated under reduced pressure and washed with CHCl 3 After filtration, the filter cake was washed with CHCl 3 (3×100 mL). The filtrate was washed with CHCl 3 (3×200 mL) extraction. The combined organic layers were washed with Na 2 S 2 O 3 (300 mL) and brine (2×150 mL), washed with anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (16% EtOAc / petroleum ether) to give the product as a solid (6.6 g, 32% yield). LCMS (ESI): C 25 H 27 F 3 IN 2 O 3 m / z [M+H] calculated: 667.03; found: 668.7

[0746] Step 3: Synthesis of 3-(5-bromo-2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate

[0747] To a stirred mixture of (S)-3-(5-bromo-2-(5-iodo-2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate (1.4 g, 2.098 mmol) and (R)-octahydro-2H-pyrido[1,2-a]pyrazine (353.04 mg, 2.518 mmol) in toluene (10 mL) was added Cs 2 CO 3 (3076.05 mg, 9.441 mmol), BINAP (65.32 mg, 0.105 mmol) and Pd(OAc) 2 (47.10 mg, 0.210 mmol). The resulting mixture was stirred at 90 °C overnight under argon atmosphere. 2The reaction was quenched with 10% HO (100 mL). The resulting mixture was extracted with DCM (3×100 mL). The combined organic layers were washed with H 2 O (3 × 100 mL), washed with anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (5% MeOH / DCM) to give the product as an oil (1 g, 49% yield). LCMS (ESI): C 33 H 42 F 3 N 4 O 3 m / z [M+H] calculated: 679.25; found: 679.5

[0748] Step 4: Synthesis of (S)-1-((S)-3-(3-(3-(3-acetoxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester

[0749] To a stirred mixture of 3-(5-bromo-2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate (1 g, 1.471 mmol) and (S)-methyl 1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylate (913.62 mg, 1.765 mmol) in toluene (9 mL) was added dioxane (6 mL), H 2 O (3 mL), K 3 PO 4 (780.82 mg, 3.678 mmol) and Pd(dtbpf)Cl 2 (95.90 mg, 0.147 mmol), the resulting mixture was stirred at 70°C for 2 hours under a nitrogen atmosphere. The mixture was washed with NaHCO 3 The saturated aqueous solution was basified to pH 8. The resulting mixture was extracted with DCM (3×30 mL). The combined organic layers were washed with H 2 O (3 × 30 mL), washed with anhydrous Na 2 SO 4After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (20% EtOAc / petroleum ether) to give the product as a solid (1.2 g, 74% yield). LCMS (ESI): C 53 H 70 F 3 N 7 O 8 m / z [M+H] calculated: 990.53; found: 990.8

[0750] Step 5: Synthesis of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylic acid

[0751] To a stirred mixture of (S)-methyl 1-((S)-3-(3-(3-(3-acetoxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (1.2 g, 1.212 mmol) and LiOH (252 mg, 10.523 mmol) in THF (6 mL) was added H in portions at 0 °C. 2 O (6 mL). The resulting mixture was stirred at 0°C overnight. The mixture was acidified to pH 7 with 1N HCl (aq). The aqueous layer was extracted with DCM (3 x 30 mL). The combined organic layers were washed with H 2 O (30 mL), washed with anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure to obtain the product as a solid (1.2 g, 84% yield). LCMS (ESI) C 50 H 66 F 3 N 7 O 7 m / z [M+H] calculated: 934.51; found: 935.0

[0752] Step 6: Synthesis 3S,4S)-12-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-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-tert-butyl 8-oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzheterocycloundecanoyl-4-yl)carbamate

[0753] To a stirred mixture of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylic acid (1.2 g, 1.285 mmol) and DIPEA (7.83 mL, 44.975 mmol) in DCM (100 mL) was added HOBt (0.87 g, 6.425 mmol) and EDCI·HCl (5.58 g, 35.980 mmol) in portions at 0° C. The resulting mixture was stirred overnight at 0° C. The mixture was diluted with DCM (30 mL). The combined organic layers were washed with H 2 O (3 × 30 mL), washed with anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (5% MeOH / DCM) to give the product as a solid (850 mg, 65% yield). LCMS (ESI): C 50 H 64 F 3 N 7 O 6 m / z [M+H] calcd: 916.49; found: 917.0

[0754] Step 7: Synthesis (6 3 S,4S)-4-amino-12-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-10,10-dimethyl-1 1 -(2,2,2-trifluoroethyl)-61 ,6 2 ,6 3 ,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzene heterocycle undecanoic acid-5,7-dione

[0755] At 0℃, ((6 3 S,4S)-12-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-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 stirred mixture of tert-butyl H-8-oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzheterocycloundecyl-4-yl)carbamate (1000 mg, 1.092 mmol) in DCM (4 mL) was added TFA (4 mL). The resulting mixture was stirred at 0° C. for 1 hour under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was washed with NaHCO 3 The saturated aqueous solution was basified to pH 8. The aqueous layer was extracted with DCM (3×30 mL). The combined organic layers were washed with anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure to obtain the product as a solid (800 mg, 80% yield). LCMS (ESI) C 45 H 56 F 3 N 7 O 4 m / z [M+H] Calculated: 816.44 Found: 816.6

[0756] Intermediate 12: Synthesis (6 3 S,4S)-4-amino-1 1 -Ethyl-1 2 -(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6 1 ,6 2 ,6 3,6 4 ,6 5 ,6 6 -Hexahydro-1 1 H-8-Oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzene heterocycle undecanoic acid-5,7-dione

[0757]

[0758] Step 1: Synthesis of (S)-1-((S)-3-(3-(3-(3-acetoxy-2,2-dimethylpropyl)-1-ethyl-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester

[0759] Under argon atmosphere, 3-(5-bromo-1-ethyl-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropyl acetate (14.2 g, 22.625 mmol), (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylic acid methyl ester (17.56 g, 33.938 mmol), and H-containing 2 O (30 mL) in dioxane (150 mL), Pd(dtbpf)Cl 2 (1.47 g, 2.263 mmol). The resulting mixture was stirred at 65 °C for 3 hours and then cooled to room temperature. The mixture was filtered and the filter cake was washed with EtOAc (2×200 mL). The filtrate was concentrated under reduced pressure and then extracted with EtOAc (2×200 mL). The combined organic layers were washed with brine (2×250 mL) and purified by anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (3→4% MeOH / DCM) to give the product as a solid (17.2 g, 81% yield). LCMS (ESI): C 52 H 71 N 7 O 9 m / z [M+H] calculated: 938.54; found: 938.8

[0760] Step 2: Synthesis of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(1-ethyl-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylic acid

[0761] To a 250 mL 3-necked round-bottom flask was added (S)-methyl 1-((S)-3-(3-(3-(3-acetoxy-2,2-dimethylpropyl)-1-ethyl-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (17.2 g, 18.33 mmol) and THF (175 mL) at room temperature. At 0° C., LiOH (1.88 g, 78.343 mmol) was added in portions to H 2 O (78.34mL, 4348.526mmol) in a stirred mixture. The resulting mixture was stirred at room temperature for 16 hours. The mixture was acidified to pH 5 with 1N HCl. The resulting mixture was extracted with EtOAc (2×200mL). The combined organic layers were washed with brine (2×200mL) and washed with anhydrous Na 2 SO 4 After filtration, the filtrate was concentrated under reduced pressure. The crude mixture (17 g, crude product) in a solid state was used directly in the next step without further purification. LCMS (ESI) C 49 H 67 N 7 O 8 m / z [M+H] calcd: 882.51; found: 882.8.

[0762] Step 3: Synthesis ((6 3 S,4S)-1 1 -Ethyl-1 2 -(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-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-tert-Butyl hexahydro-11H-8-oxa-1(5,3)-indol-6(1,3)-pyridazin-2(1,3)-benzheterocycloundecyl-4-yl)carbamate

[0763] To a 5 L 3-necked round bottom flask was added (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(1-ethyl-2-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylic acid (16.8 g, 19.045 mmol) and DCM (2.52 L) at ro...

Claims

1. A compound or a pharmaceutically acceptable salt thereof having a structure of Formula I: wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene; X 1 , X 2 and X 3 Each independently selected from CH2, CHF, CF2, C=O or O; m is 1 or 2; n is 0 or 1; R 1 is hydrogen, optionally substituted C1-C6 heteroalkyl, or optionally substituted 3- to 10-membered heterocycloalkyl; R 2 is an optionally substituted C1-C6 alkyl group; and R 3 is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, an optionally substituted 3- to 6-membered cycloalkyl group, or an optionally substituted heterocycloalkyl group, and wherein each hydrogen is independently optionally isotopically enriched with deuterium.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 is hydrogen or optionally substituted 3- to 10-membered heterocycloalkyl.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 is an optionally substituted 3- to 10-membered heterocycloalkyl group.

4. The compound according to claim 3 or a pharmaceutically acceptable salt thereof, wherein R 1 yes: 5 . The compound according to claim 1 , or a pharmaceutically acceptable salt thereof, wherein m is 1. 6 . The compound or pharmaceutically acceptable salt thereof according to claim 1 , wherein n is 1.

7. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein X 1 , X 2 and X 3 Each is CH2.

8. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, having a structure of Formula II:

9. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, having a structure of Formula V:

10. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, having a structure of Formula VI:

11. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, having a structure of Formula VII:

12. A compound as described in any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, wherein A is an optionally substituted thiazole-diyl, an optionally substituted oxazole-diyl, an optionally substituted morpholine-diyl, an optionally substituted pyrrolidine-diyl, an optionally substituted piperidine-diyl, or an optionally substituted phenylene.

13. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, wherein A is an optionally substituted 5- to 10-membered heteroarylene.

14. The compound of claim 13 or a pharmaceutically acceptable salt thereof, wherein A is:

15. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, wherein A is an optionally substituted phenylene group.

16. The compound of claim 15 or a pharmaceutically acceptable salt thereof, wherein A is:

17. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene group.

18. The compound of claim 17 or a pharmaceutically acceptable salt thereof, wherein A is selected from the following or stereoisomers thereof:

19. A compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, wherein R 2 yes:

20. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 19, wherein R 3 It is an optionally substituted C1-C6 alkyl group or an optionally substituted 3- to 6-membered cycloalkyl group.

21. A compound according to any one of claims 1 to 20, or a pharmaceutically acceptable salt thereof, wherein R 3 It is an optionally substituted C1-C6 alkyl group.

22. The compound of claim 21 or a pharmaceutically acceptable salt thereof, wherein R 3 yes:

23. A compound as claimed in any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof, wherein R 3 is or is an optionally substituted 3- to 6-membered cycloalkyl.

24. The compound of claim 23 or a pharmaceutically acceptable salt thereof, wherein R 3 yes:

25. A compound as described in any one of claims 1 to 24, or a pharmaceutically acceptable salt thereof, wherein the compound is not a compound of Table 3.

26. A compound or a pharmaceutically acceptable salt thereof having the structure of a compound of Table 1 or Table 3.

27. A pharmaceutical composition comprising the compound according to any one of claims 1 to 26 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

28. A method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 26, or a pharmaceutical composition according to claim 27.

29. The method of claim 28, wherein the cancer is pancreatic cancer, non-small cell lung cancer, colorectal cancer, or endometrial cancer.

30. The method of claim 28 or 29, wherein the cancer comprises a Ras mutation.

31. The method of claim 30, wherein the Ras mutation is K-Ras G12D or K-G13D.

32. A method for treating a Ras protein-related disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 26, or a pharmaceutical composition according to claim 27.

33. A method for inhibiting Ras protein in a cell, the method comprising contacting the cell with an effective amount of the compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 26 or the pharmaceutical composition according to claim 27.

34. The method of claim 32 or 33, wherein the Ras protein is K-Ras G12D or K-Ras G13D.

35. The method of claim 33 or 34, wherein the cell is a cancer cell.

36. The method of claim 35, wherein the cancer cell is a pancreatic cancer cell, a non-small cell lung cancer cell, a colorectal cancer cell, or an endometrial cell.

37. The method of any one of claims 28 to 36, wherein the method further comprises administering an additional anti-cancer therapy.

38. The method of claim 37, wherein the additional anti-cancer therapy is an EGFR inhibitor, a second Ras inhibitor, a SHP2 inhibitor, a SOS1 inhibitor, a Raf inhibitor, a MEK inhibitor, an ERK inhibitor, a PI3K inhibitor, a PTEN inhibitor, an AKT inhibitor, an mTORC1 inhibitor, a BRAF inhibitor, a PD-L1 inhibitor, a PD-1 inhibitor, a CDK4 / 6 inhibitor, a HER2 inhibitor, or a combination thereof.

39. A conjugate or a salt thereof comprising a structure of formula III: M-P 1 Formula III Where P 1 is a monovalent organic fraction; and M has the structure of Formula IV: wherein A is an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 6-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene; X 1 , X 2 and X 3 Each independently selected from CH2, CHF, CF2, C=O or O; m is 1 or 2; n is 0 or 1; R 1 is hydrogen, optionally substituted C1-C6 heteroalkyl, or optionally substituted 3- to 10-membered heterocycloalkyl; R 2 is an optionally substituted C1-C6 alkyl group; and R 3 is an optionally substituted C1-C6 alkyl group, an optionally substituted 3- to 6-membered cycloalkyl group, an optionally substituted C1-C6 heteroalkyl group or an optionally substituted heterocycloalkyl group, and wherein each hydrogen of Formula IV is independently optionally isotopically enriched with deuterium.

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