HK40057874A - Inhibitor of indoleamine-2,3-dioxygenase (ido) - Google Patents

Description

Indoleamine-2,3-dioxygenase (IDO) inhibitors

Related applications

This application is a divisional application of Chinese invention patent application No. 201780009388.3, filed with the China Intellectual Property Office on February 8, 2017, with the invention name “Indoleamine-2,3-dioxygenase (IDO) inhibitors”.

Background Art

Indoleamine 2,3-dioxygenase (IDO), such as indoleamine 2,3-dioxygenase 1 (IDO1), is a family of heme-containing enzymes that catalyze the degradation of the essential amino acid L-tryptophan to N-formylkynurenine. They play an important role in the initiation and rate-limiting step of tryptophan degradation.

IDO (e.g., IDO1), an enzyme induced by IFNγ, has been reported to be one of the central regulators of immune responses in various physiological and pathological settings. IDO leads to immunosuppression by degrading tryptophan in the tumor microenvironment. (Selvan et al., Curr. Cancer Drug Targets, 2015; Baren and Eynde Cancer Immunology Research, 2015). Overexpression of IDO has been observed in various tumors (e.g., colorectal cancer, ovarian cancer, and breast cancer), which is thought to enable tumor cells to evade immune surveillance. [Godin-Ethier et al., Clinical Cancer Research, 2011 Nov 15;17(22):6985-91]. Treg cells have also been found to regulate IDO-mediated tryptophan catabolism in dendritic cells. (Fallarino et al., Nature Immunology 2003). In addition, IDO has been associated with other diseases, such as viral infections and Alzheimer's disease. Therefore, IDO is a promising target for cancer (e.g., cancer immunotherapy) as well as other diseases (e.g., infectious diseases and Alzheimer's disease).

Summary of the Invention

The present disclosure provides compounds, such as compounds of formula (I), which inhibit IDO, such as IDO1 and thus inhibit tryptophan catabolism and kynurenine reduction in the tumor microenvironment and surrounding lymph nodes. The compounds described herein can be used to treat proliferative diseases, such as cancer (e.g., non-small cell lung cancer, small cell lung cancer, breast cancer, prostate cancer, ovarian cancer, bladder cancer, head and neck cancer, renal cell carcinoma, pancreatic cancer, brain cancer, gastrointestinal cancer, liver cancer, leukemia, lymphoma, melanoma, multiple myeloma, Ewing's sarcoma, osteosarcoma and neuroblastoma) and infectious diseases, such as viral or bacterial infectious diseases (e.g., hepatitis and HIV). Pharmaceutical compositions, kits, methods and uses of any compound described herein are also provided.

In one aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, wherein, where valence permits, W is -O-, -S-, or a bond; Q is -C(=O)NH- or a bond; and Y is -CR ⁸ = or -N=. In addition,

R ¹ is —C(═O)OH, —C(═O)OR ¹⁰ , a substituted or unsubstituted heterocyclic group, a substituted or unsubstituted heteroaryl group, —NHSO ₂ R ⁹ , —C(═O)NHSO ₂ R ⁹ , —C(═O)NHC(═O)OR ¹⁰ , or —SO ₂ NHC(═O)R ¹⁰ ;

^R2 and ^R3 are each independently hydrogen, halogen, substituted or unsubstituted _C1 - _C6 alkyl, substituted or unsubstituted _C1 - _C6 alkoxy, or ^R2 and ^R3 are linked to form a substituted or unsubstituted 3- to 8-membered carbocyclic ring or a substituted or unsubstituted 3- to 8-membered heterocyclic ring;

^R4 and ^R5 are each independently hydrogen, substituted or unsubstituted _C1 - _C6 alkyl, substituted or unsubstituted _C2 - _C6 alkenyl, substituted or unsubstituted C5- _C8 cycloalkenyl, substituted or unsubstituted C2- _C10 alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted _{C1 - C6} alkoxy, substituted or unsubstituted _C3 - _C8 cycloalkyl, substituted or unsubstituted 3- to ₁₂ -membered heterocyclyl (e.g., heterocycloalkyl), substituted or unsubstituted 5- to 6-membered monocyclic heteroaryl, substituted or unsubstituted 8- to 10-membered bicyclic heteroaryl, or arylsulfonyl; or ^R4 and ^R5, together with the N to which they are attached, are linked to form an optionally substituted heterocyclyl, which may be monocyclic or bicyclic.

R ⁶ is substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₃ -C ₈ cycloalkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, or substituted or unsubstituted C ₅ -C ₈ cycloalkenyl, substituted or unsubstituted aryl, substituted or unsubstituted 4- to 7-membered monocyclic heterocyclyl (e.g., heterocycloalkyl), substituted or unsubstituted 7- to 10-membered bicyclic heterocyclyl, substituted or unsubstituted 5- to 6-membered monocyclic heteroaryl, substituted or unsubstituted 8- to 10-membered bicyclic heteroaryl, substituted or unsubstituted C ₁ -C ₆ alkoxy, substituted or unsubstituted aryloxy, or -C(═O)R ⁷ ;

R ⁷ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, or substituted or unsubstituted aryl;

R ⁸ is independently hydrogen, halogen, -CN, -OH, substituted or unsubstituted C ₁ -C ₆ alkyl, or substituted or unsubstituted C ₁ -C ₆ alkoxy; and

^R9 and ^R10 are each independently hydrogen, substituted or unsubstituted _C1 - _C6 alkyl, substituted or unsubstituted _C2 - _C6 alkenyl; or a pharmaceutically acceptable salt, wherein ^R1 , ^R2 , ^R3 , ^R4 , ^R5 , ^R6 , ^R7 , ^R8 , ^R9 , ^R10 , Y and Q are as defined herein.

In certain embodiments, R ¹ is —C(═O)OH, substituted or unsubstituted heterocyclyl, —NHSO ₂ R ⁹ , —C(═O)NHSO ₂ R ⁹ , —C(═O)NHC(═O)OR ¹⁰ , or —SO ₂ NHC(═O)R ¹⁰ .

In certain embodiments, ^R4 and ^R5 are each independently hydrogen, substituted or unsubstituted _C1 - _C6 alkyl, substituted or unsubstituted _C2 - _C6 alkenyl, substituted or unsubstituted _C5 - _C8 cycloalkenyl, substituted or unsubstituted _C2 - _C10 alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted _C1 - _C6 alkoxy, substituted or unsubstituted _C3 - _C8 cycloalkyl, substituted or unsubstituted 3- to 12-membered heterocyclyl (e.g., heterocycloalkyl), substituted or unsubstituted 5- to 6-membered monocyclic heteroaryl, substituted or unsubstituted 8- to 10-membered bicyclic heteroaryl, substituted or unsubstituted aryl, or arylsulfonyl.

In certain embodiments, the compound of formula (I) is a compound of formula (II):

or a pharmaceutically acceptable salt, wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , Y and Q are as defined herein.

Exemplary compounds of formula (II) include, but are not limited to:

and pharmaceutically acceptable salts.

In certain embodiments, the compound of formula (I) is a compound of formula (III):

or a pharmaceutically acceptable salt, wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , Y and Q are as defined herein.

Exemplary compounds of formula (III) also include, but are not limited to:

and pharmaceutically acceptable salts.

In certain embodiments, the compound of formula (I) is a compound of formula (IV):

or a pharmaceutically acceptable salt, wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , Y and Q are as defined herein.

Exemplary compounds of formula (IV) include, but are not limited to:

and pharmaceutically acceptable salts.

In certain embodiments, the compound of formula (I) is a compound of formula (V):

or a pharmaceutically acceptable salt, wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , W and Y are as defined herein.

Exemplary compounds of formula (V) include, but are not limited to:

and pharmaceutically acceptable salts.

In another aspect, the present disclosure provides pharmaceutical compositions comprising one or more compounds described herein and a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical compositions described herein comprise an effective amount of a compound described herein for inhibiting the catabolism of IDO and tryptophan, resulting in reduced kynurenine levels. The effective amount described herein can be a therapeutically effective amount or a prophylactically effective amount.

In yet another aspect, the present disclosure provides methods for treating a disease associated with IDO (eg, cancer or an infectious disease), comprising administering to a subject in need of such treatment an effective amount of any of the pharmaceutical compositions described herein.

In certain embodiments, target cancers include, but are not limited to, non-small cell lung cancer, small cell lung cancer, breast cancer, prostate cancer, ovarian cancer, bladder cancer, head and neck cancer, renal cell carcinoma, pancreatic cancer, brain cancer, gastrointestinal cancer, liver cancer, leukemia, lymphoma, melanoma, multiple myeloma, Ewing's sarcoma, osteosarcoma, and neuroblastoma.

In certain embodiments, the subject treated is a mammal (eg, a human or non-human mammal).

In certain embodiments, the present disclosure provides for the use of an IDO inhibitory compound as described herein in combination therapy with another anti-cancer therapy for a cancer patient, including but not limited to immunotherapy, radiation therapy, surgery, chemotherapy, and cell therapy. In some instances, the other anti-cancer therapy involves the use of one or more anti-cancer agents.

Another aspect of the present disclosure relates to a kit comprising a container containing a compound as described herein or a pharmaceutical composition thereof. The kits described herein may include a single dose or multiple doses of the compound or pharmaceutical composition. The kits may be used in the methods of the present disclosure. In certain embodiments, the kits further include instructions for use of the compound or pharmaceutical composition.

In yet another aspect, the present disclosure provides uses of the compounds and pharmaceutical compositions described herein for treating a proliferative disease (such as the cancers described herein) and/or for preparing a medicament for treating a target disease.

The details of one or more embodiments of the present disclosure are set forth herein. Other features, objects, and advantages of the present disclosure will become apparent from the detailed description, examples, and claims.

BRIEF DESCRIPTION OF THE DRAWINGS

Non-limiting embodiments of the present disclosure will be described by way of example with reference to the accompanying drawings, which are schematic and not intended to be drawn to scale. In the accompanying drawings, each identical or nearly identical component shown is generally represented by a single numeral. For clarity, not every component is labeled in every drawing, and not every component of every embodiment of the invention is shown, where illustration is not necessary for a person of ordinary skill in the art to understand the present invention.

FIG1 is a graph showing that Compound 9 and INCB-24360 inhibit kynurenine production in SKOV-3 cells.

FIG2 is a graph showing LPS-induced plasma kynurenine levels in mice in the presence or absence of Compound 9. FIG.

Figure 3 includes graphs showing the reduction of kynurenine in human whole blood samples after compound treatment. Panel A: Percent inhibition of the kynurenine/tryptophan ratio for compound 84 and compound INCB-24360, respectively, as a function of each compound's concentration. Panel B: Percent inhibition of kynurenine for compound 84 and compound INCB-24360, respectively, as a function of each compound's concentration.

Figure 4 is a graph showing that IDO inhibitors increase IFN-γ production in a co-culture of T cells and HeLa cells, indicating that IDO inhibitors activate T cells. The _EC50s for INCB-24360 and Compound 120 in this assay are 41 nM and 9.1 nM, respectively.

definition

Definitions of specific functional groups and chemical terms are described in more detail below.The chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th ed., inside cover, and specific functional groups are generally defined as described therein. In addition, general principles of organic chemistry as well as specific functional moieties and reactivities are described in the following books: Thomas Sorrell, Organic Chemistry, University Science Books, Sausalito, 1999; Smith and March, March's Advanced Organic Chemistry, ^5th Edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; and Carruthers, Some Modern Methods in Organic Synthesis, 3rd Edition, Cambridge University Press, Cambridge, 1987. Methods of Organic Synthesis, ^3rd Edition, Cambridge University Press, Cambridge, 1987). The present invention is not intended to be limited in any way by the exemplary lists of substituents described herein.

The compounds described herein may contain one or more asymmetric centers and may therefore exist in various isomeric forms, such as enantiomers and/or diastereomers. For example, the compounds described herein may be in the form of individual enantiomers, diastereomers, or geometric isomers, or may be in the form of mixtures of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers may be separated from the mixture by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferably, isomers may be prepared by asymmetric synthesis. See, e.g., Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables of Resolving Agents and Optical Resolutions, p. 268 (E. L. Eliel, ed., University of Notre Dame Press, Notre Dame, IN 1972). The present disclosure further encompasses the compounds described herein as individual isomers substantially free of other isomers, or as mixtures of various isomers.

When a range of values is listed, it is intended to encompass every value and subrange within that range. For example, “ _C1-6 ” is intended to encompass _C1 , _C2 , _C3 , _C4 , _C5 , C6, _C1-6 , _C1-5 , _C1-4 , _C1-3 , _C1-2 , _C2-6 , _C2-5 , _C2-4 , _C2-3 , _C3-6 , _C3-5 , _{C3-4 , C4-6 , C4-5} , and _C5-6 .

The term "aliphatic" includes saturated and unsaturated straight chain (i.e., unbranched), branched, acyclic, cyclic or polycyclic aliphatic hydrocarbons, which are optionally substituted by one or more functional groups. As will be understood by those skilled in the art, "aliphatic" is intended to include, but is not limited to, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl and cycloalkynyl moieties in this article. Therefore, the term "alkyl" includes straight chain, branched and cyclic alkyl. Similar conventions apply to other general terms, such as "alkenyl", "alkynyl" etc. In addition, the terms "alkyl", "alkenyl", "alkynyl" etc. include substituted and unsubstituted groups. In certain embodiments, "low alkyl" is used to refer to those alkyl groups (cyclic, acyclic, substituted, unsubstituted, branched or unbranched) having 1-6 carbon atoms.

In certain embodiments, the alkyl, alkenyl and alkynyl used in the present disclosure contain 1-20 aliphatic carbon atoms. In certain other embodiments, the alkyl, alkenyl and alkynyl used in the present disclosure contain 1-10 aliphatic carbon atoms. In other embodiments, the alkyl, alkenyl and alkynyl used in the present disclosure contain 1-8 aliphatic carbon atoms. In other embodiments, the alkyl, alkenyl and alkynyl used in the present disclosure contain 1-6 aliphatic carbon atoms. In still other embodiments, the alkyl, alkenyl and alkynyl used in the present disclosure contain 1-4 carbon atoms. Thus, exemplary aliphatic groups include, but are not limited to, for example, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, -CH ₂ -cyclopropyl, vinyl, allyl, n-butyl, sec-butyl, isobutyl, tert-butyl, cyclobutyl, -CH ₂ -cyclobutyl, n-pentyl, sec-pentyl, isopentyl, tert-pentyl, cyclopentyl, -CH ₂ -cyclopentyl, n-hexyl, sec-hexyl, cyclohexyl, -CH ₂ -cyclohexyl moieties, etc., which may further carry one or more substituents. Alkenyl groups include, but are not limited to, for example, vinyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, etc. Representative alkynyl groups include, but are not limited to, ethynyl, 2-propynyl (propargyl), 1-propynyl, etc.

The term "alkyl" refers to a group of straight or branched saturated hydrocarbon groups having 1 to 10 carbon atoms ("C _1-10 alkyl"). In some embodiments, the alkyl group has 1 to 9 carbon atoms ("C _1-9 alkyl"). In some embodiments, the alkyl group has 1 to 8 carbon atoms ("C _1-8 alkyl"). In some embodiments, the alkyl group has 1 to 7 carbon atoms ("C _1-7 alkyl"). In some embodiments, the alkyl group has 1 to 6 carbon atoms ("C _1-6 alkyl"). In some embodiments, the alkyl group has 1 to 5 carbon atoms ("C _1-5 alkyl"). In some embodiments, the alkyl group has 1 to 4 carbon atoms ("C _1-4 alkyl"). In some embodiments, the alkyl group has 1 to 3 carbon atoms ("C _1-3 alkyl"). In some embodiments, the alkyl group has 1 to 2 carbon atoms ("C _1-2 alkyl"). In some embodiments, the alkyl group has 1 carbon atom ("C ₁ alkyl"). In some embodiments, an alkyl group has 2 to 6 carbon atoms (" _C2-6 alkyl"). Examples of _C1-6 alkyl groups include methyl ( _C1 ), ethyl ( _C2 ), propyl ( _C3 ) (e.g., n-propyl, isopropyl), butyl ( _C4 ) (e.g., n-butyl, tert-butyl, sec-butyl, isobutyl), pentyl ( _C5 ) (e.g., n-pentyl, 3-pentyl, neopentyl, 3-methyl-2-butyl, tert-pentyl), and hexyl ( _C6 ) (e.g., n-hexyl). Additional examples of alkyl groups include n-heptyl ( _C7 ), n-octyl ( _C8 ), and the like. Unless otherwise specified, each instance of an alkyl group is independently unsubstituted ("unsubstituted alkyl") or substituted ("substituted alkyl") with one or more substituents (e.g., halogen, such as F). In certain embodiments, an alkyl group is an unsubstituted _C1-10 alkyl group (e.g., an unsubstituted _C1-6 alkyl group, such as _-CH3 ). In certain embodiments, the alkyl group is a substituted C _1-10 alkyl group (eg, a substituted C _1-6 alkyl group, such as -CF ₃ ).

“Alkenyl” refers to a group of straight or branched hydrocarbon groups having 2 to 20 carbon atoms, one or more carbon-carbon double bonds, and no triple bonds (“C _2-20 alkenyl”). In some embodiments, the alkenyl group has 2 to 10 carbon atoms (“C _2-10 alkenyl”). In some embodiments, the alkenyl group has 2 to 9 carbon atoms (“C _2-9 alkenyl”). In some embodiments, the alkenyl group has 2 to 8 carbon atoms (“C _2-8 alkenyl”). In some embodiments, the alkenyl group has 2 to 7 carbon atoms (“C _2-7 alkenyl”). In some embodiments, the alkenyl group has 2 to 6 carbon atoms (“C _2-6 alkenyl”). In some embodiments, the alkenyl group has 2 to 5 carbon atoms (“C _2-5 alkenyl”). In some embodiments, the alkenyl group has 2 to 4 carbon atoms (“C _2-4 alkenyl”). In some embodiments, the alkenyl group has 2 to 3 carbon atoms (“C _2-3 alkenyl”). In some embodiments, an alkenyl group has 2 carbon atoms (" _C2 alkenyl"). The one or more carbon-carbon double bonds can be internal (e.g., in 2-butenyl) or terminal (e.g., in 1-butenyl). Examples of _C2-4 alkenyl groups include ethenyl ( _C2 ), 1-propenyl ( _C3 ), 2-propenyl ( _C3 ), 1-butenyl ( _C4 ), 2-butenyl ( _C4 ), butadienyl ( _C4 ), and the like. Examples of _C2-6 alkenyl groups include the aforementioned _C2-4 alkenyl groups as well as pentenyl ( _C5 ), pentadienyl ( _C5 ), hexenyl ( _C6 ), and the like. Additional examples of alkenyl groups include heptenyl ( _C7 ), octenyl ( _C8 ), octatrienyl ( _C8 ), and the like. Unless otherwise specified, each instance of an alkenyl group is independently optionally substituted, i.e., unsubstituted (an "unsubstituted alkenyl group") or substituted (a "substituted alkenyl group") with one or more substituents. In certain embodiments, an alkenyl group is an unsubstituted _C2-10 alkenyl group. In certain embodiments, an alkenyl group is a substituted _C2-10 alkenyl group. In an alkenyl group, a C=C double bond (e.g., -CH= _CHCH3 or) where the stereochemistry is not specified can be an (E)- or (Z)-double bond.

“Alkynyl” refers to a group of straight or branched hydrocarbon groups having 2 to 20 carbon atoms, one or more carbon-carbon triple bonds, and optionally one or more double bonds (“C _2-20 alkynyl”). In some embodiments, the alkynyl group has 2 to 10 carbon atoms (“C _2-10 alkynyl”). In some embodiments, the alkynyl group has 2 to 9 carbon atoms (“C _2-9 alkynyl”). In some embodiments, the alkynyl group has 2 to 8 carbon atoms (“C _2-8 alkynyl”). In some embodiments, the alkynyl group has 2 to 7 carbon atoms (“C _2-7 alkynyl”). In some embodiments, the alkynyl group has 2 to 6 carbon atoms (“C _2-6 alkynyl”). In some embodiments, the alkynyl group has 2 to 5 carbon atoms (“C _2-5 alkynyl”). In some embodiments, the alkynyl group has 2 to 4 carbon atoms (“C _2-4 alkynyl”). In some embodiments, the alkynyl group has 2 to 3 carbon atoms (“C _2-3 alkynyl”). In some embodiments, an alkynyl group has 2 carbon atoms (" _C2 alkynyl"). The one or more carbon-carbon triple bonds can be internal (e.g., in 2-butynyl) or terminal (e.g., in 1-butynyl). Examples of _C2-4 alkynyl groups include, but are not limited to, ethynyl ( _C2 ), 1-propynyl ( _C3 ), 2-propynyl ( _C3 ), 1-butynyl ( _C4 ), 2-butynyl ( _C4 ), and the like. Examples of _C2-6 alkenyl groups include the aforementioned _C2-4 alkynyl groups as well as pentynyl ( _C5 ), hexynyl ( _C6 ), and the like. Additional examples of alkynyl groups include heptynyl ( _C7 ), octynyl ( _C8 ), and the like. Unless otherwise specified, each instance of an alkynyl group is independently optionally substituted, i.e., unsubstituted ("unsubstituted alkynyl") or substituted ("substituted alkynyl") with one or more substituents. In certain embodiments, an alkynyl group is an unsubstituted _C2-10 alkynyl group. In certain embodiments, the alkynyl group is a substituted C _2-10 alkynyl group.

"Carbocyclyl" or "carbocyclic" refers to a group of non-aromatic cycloalkyl groups having 3 to 10 ring carbon atoms (" _C3-10 carbocyclyl") and zero heteroatoms in a non-aromatic ring system. In some embodiments, the carbocyclyl group has 3 to 8 ring carbon atoms (" _C3-8 carbocyclyl"). In some embodiments, the carbocyclyl group has 3 to 6 ring carbon atoms (" _C3-6 carbocyclyl"). In some embodiments, the carbocyclyl group has 3 to 6 ring carbon atoms (" _C3-6 carbocyclyl"). In some embodiments, the carbocyclyl group has 5 to 10 ring carbon atoms (" _C5-10 carbocyclyl"). Exemplary _C3-6 carbocyclyl groups include, but are not limited to, cyclopropyl ( _C3 ), cyclopropenyl (C3), cyclobutyl ( _C4 ), cyclobutenyl ( _C4 ), cyclopentyl ( _C5 ), cyclopentenyl ( _C5 ), cyclohexyl ( _C6 ), cyclohexenyl ( _C6 ), cyclohexadienyl ( _C6 ), etc. Exemplary _C3-8 carbocyclyl groups include, but are not limited to, the above-mentioned _C3-6 carbocyclyl groups as well as cycloheptyl ( _C7 ), cycloheptenyl ( _C7 ), cycloheptadienyl ( _C7 ), cycloheptatrienyl ( _C7 ), cyclooctyl ( _C8 ), cyclooctenyl ( _{C8 )} , bicyclo[2.2.1]heptyl ( _C7 ), bicyclo[2.2.2]octyl ( _C8 ), etc. Exemplary _C3-10 carbocyclyls include, but are not limited to, the aforementioned _C3-8 carbocyclyls as well as cyclononyl (C9), cyclononenyl ( _C9 ), cyclodecyl ( _C10 ), cyclodecenyl ( _C10 ), octahydro-1H-indenyl ( _C9 ), decahydronaphthyl ( _C10 ), spiro[4.5]decyl ( _C10 ), and the like. As shown in the foregoing examples, in certain embodiments, the carbocyclyl is a monocyclic ring ("monocyclic carbocyclyl") or contains a fused, bridged, or spirocyclic ring system, such as a bicyclic ring system (" _bicyclic carbocyclyl") and may be saturated or partially unsaturated. "Carbocyclyl" also includes ring systems in which the point of attachment of a carbocyclic ring as defined above to one or more aryl or heteroaryl groups is fused to the carbocyclic ring, and in this case, the carbon number continues to refer to the number of carbons in the carbocyclic ring system. Unless otherwise specified, each instance of a carbocyclyl group is independently optionally substituted, i.e., unsubstituted ("unsubstituted carbocyclyl") or substituted ("substituted carbocyclyl") with one or more substituents. In certain embodiments, a carbocyclyl group is an unsubstituted _C3-10 carbocyclyl group. In certain embodiments, a carbocyclyl group is a substituted _C3-10 carbocyclyl group.

In some embodiments, a “carbocyclyl” is a monocyclic saturated carbocyclyl having 3 to 10 ring carbon atoms (“C _3-10 cycloalkyl”). In some embodiments, a cycloalkyl has 3 to 8 ring carbon atoms (“C _3-8 cycloalkyl”). In some embodiments, a cycloalkyl has 3 to 6 ring carbon atoms (“C _3-6 cycloalkyl”). In some embodiments, a cycloalkyl has 5 to 6 ring carbon atoms (“C _5-6 cycloalkyl”). In some embodiments, a cycloalkyl has 5 to 10 ring carbon atoms (“C _5-10 cycloalkyl”). Examples of C _5-6 cycloalkyls include cyclopentyl (C ₅ ) and cyclohexyl (C ₆ ). Examples of C _3-6 cycloalkyls include the aforementioned C _5-6 cycloalkyls as well as cyclopropyl (C ₃ ) and cyclobutyl (C ₄ ). Examples of C _3-8 cycloalkyls include the aforementioned C _3-6 cycloalkyls as well as cycloheptyl (C ₇ ) and cyclooctyl (C ₈ ). Unless otherwise specified, each instance of a cycloalkyl group is independently unsubstituted (an "unsubstituted cycloalkyl group") or substituted (a "substituted cycloalkyl group") with one or more substituents. In certain embodiments, a cycloalkyl group is an unsubstituted _C3-10 cycloalkyl group. In certain embodiments, a cycloalkyl group is a substituted _C3-10 cycloalkyl group.

"Heterocyclyl" or "heterocyclic" refers to a group of a 3 to 10 membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon ("3-10 membered heterocyclyl"). In heterocyclyl groups containing one or more nitrogen atoms, the point of attachment may be a carbon atom or a nitrogen atom as long as valence permits. A heterocyclyl group may be a monocyclic ring ("monocyclic heterocyclyl") or a fused ring, bridged ring or spirocyclic ring system, such as a bicyclic ring system ("bicyclic heterocyclyl"), and may be saturated or partially unsaturated. A heterocyclyl bicyclic ring system may contain one or more heteroatoms in one or both rings. "Heterocyclyl" also includes ring systems in which the point of attachment of a heterocycle as defined above to one or more carbocyclyl groups is fused to a carbocyclyl or heterocycle, or a ring system in which the point of attachment of a heterocycle as defined above to one or more aryl or heteroaryl groups is fused to a heterocycle, and in such cases, the number of ring members continues to refer to the number of ring members in the heterocycle system. Unless otherwise specified, each instance of heterocyclyl is independently optionally substituted, i.e., unsubstituted (an "unsubstituted heterocyclyl") or substituted (a "substituted heterocyclyl") with one or more substituents. In certain embodiments, heterocyclyl is an unsubstituted 3-10 membered heterocyclyl. In certain embodiments, heterocyclyl is a substituted 3-10 membered heterocyclyl.

In some embodiments, a heterocyclyl group is a 5-10 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon (“5-10 membered heterocyclyl”). In some embodiments, a heterocyclyl group is a 5-8 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heterocyclyl”). In some embodiments, a heterocyclyl group is a 5-6 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heterocyclyl”). In some embodiments, a 5-6 membered heterocyclyl group has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, a 5-6 membered heterocyclyl group has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heterocyclyl has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur.

Exemplary 3-membered heterocyclic groups containing one heteroatom include, but are not limited to, azirdinyl, oxiranyl, and thiiranyl. Exemplary 4-membered heterocyclic groups containing one heteroatom include, but are not limited to, azetidinyl, oxetanyl, and thietanyl. Exemplary 5-membered heterocyclic groups containing one heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclic groups containing two heteroatoms include, but are not limited to, dioxolanyl, oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-membered heterocyclic groups containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic groups containing one heteroatom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, piperazinyl, morpholinyl, dithianyl, and dioxanyl. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, triazinyl. Exemplary 7-membered heterocyclic groups containing one heteroatom include, but are not limited to, azepanyl, oxepanyl, and thiepanyl. Exemplary 8-membered heterocyclic groups containing one heteroatom include, but are not limited to, azocanyl, oxecanyl, and thiocanyl. Exemplary groups in which a 5-membered heterocyclyl is fused to a C ₆ aryl ring (also referred to herein as a 5,6-bicyclic heterocycle) include, but are not limited to, dihydroindolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinonyl, etc. Exemplary groups in which a 6-membered heterocyclyl is fused to an aromatic ring (also referred to herein as a 6,6-bicyclic heterocycle) include, but are not limited to, tetrahydroquinolinyl, tetrahydroisoquinolinyl, etc.

"Aryl" refers to a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 π electrons shared in the cyclic array) having 6-14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system ("C _6-14 aryl"). In some embodiments, the aryl group has 6 ring carbon atoms ("C ₆ aryl"; e.g., phenyl). In some embodiments, the aryl group has 10 ring carbon atoms ("C ₁₀ aryl"; e.g., naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, the aryl group has 14 ring carbon atoms ("C ₁₄ aryl"; e.g., anthracenyl). "Aryl" also includes ring systems in which an aryl ring as defined above is fused to one or more carbocyclic or heterocyclic groups, wherein the radical or point of attachment is on the aromatic ring, and in this case, the number of carbon atoms continues to refer to the number of carbon atoms in the aromatic ring system. Unless otherwise specified, each instance of an aryl group is independently optionally substituted, i.e., unsubstituted (an "unsubstituted aryl") or substituted (a "substituted aryl") with one or more substituents. In certain embodiments, an aryl group is an unsubstituted _C6-14 aryl group. In certain embodiments, an aryl group is a substituted _C6-14 aryl group.

"Aralkyl" is a subset of alkyl and aryl and refers to an optionally substituted alkyl group substituted with an optionally substituted aryl group. In certain embodiments, the aralkyl group is an optionally substituted benzyl group. In certain embodiments, the aralkyl group is a benzyl group. In certain embodiments, the aralkyl group is an optionally substituted phenethyl group. In certain embodiments, the aralkyl group is a phenethyl group.

"Heteroaryl" refers to a group of a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 π electrons shared in a cyclic array) having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-10 membered heteroaryl"). In heteroaryl groups containing one or more nitrogen atoms, the point of attachment can be a carbon atom or a nitrogen atom as valence permits. Heteroaryl bicyclic ring systems can contain one or more heteroatoms in one or both rings. "Heteroaryl" includes ring systems in which a heteroaryl ring as defined above is fused to one or more carbocyclyl or heterocyclyl groups, wherein the point of attachment is on the heteroaryl ring, and in such cases, the number of ring members continues to refer to the number of ring members in the heteroaryl ring system. "Heteroaryl" also includes ring systems in which a heteroaryl ring as defined above is fused to one or more aryl groups, wherein the point of attachment is on the aryl or heteroaryl ring, and in this case, the number of ring members refers to the number of ring members in the fused (aryl/heteroaryl) ring system. For bicyclic heteroaryl groups in which one ring contains no heteroatoms (e.g., indolyl, quinolyl, carbazolyl, etc.), the point of attachment can be on either ring, i.e., the ring with the heteroatom (e.g., 2-indolyl) or the ring without heteroatoms (e.g., 5-indolyl).

In some embodiments, heteroaryl is a 5-10 membered aromatic ring system having ring carbon atoms provided in the aromatic ring system and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-10 membered heteroaryl”). In some embodiments, heteroaryl is a 5-8 membered aromatic ring system having ring carbon atoms provided in the aromatic ring system and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”). In some embodiments, heteroaryl is a 5-6 membered aromatic ring system having ring carbon atoms provided in the aromatic ring system and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”). In some embodiments, 5-6 membered heteroaryl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, 5-6 membered heteroaryl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl group has one ring heteroatom selected from nitrogen, oxygen, and sulfur. Unless otherwise indicated, each instance of a heteroaryl group is independently optionally substituted, i.e., unsubstituted (an "unsubstituted heteroaryl") or substituted (a "substituted heteroaryl") with one or more substituents. In certain embodiments, a heteroaryl group is an unsubstituted 5-14 membered heteroaryl group. In certain embodiments, a heteroaryl group is a substituted 5-14 membered heteroaryl group.

Exemplary 5-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyrrolyl, furanyl, and thienyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridinyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing one heteroatom include, but are not limited to, azepinyl, oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groups include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, but are not limited to, naphthyridinyl, pteridinyl, quinolyl, isoquinolyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

"Heteroaralkyl" is a subset of alkyl and heteroaryl, and refers to an optionally substituted alkyl group substituted by an optionally substituted heteroaryl group.

"Unsaturated" or "partially unsaturated" refers to a group that contains at least one double or triple bond. "Partially unsaturated" ring systems are also intended to encompass rings with multiple sites of unsaturation, but are not intended to include aromatic groups (e.g., aryl or heteroaryl groups). Likewise, "saturated" refers to groups that do not contain double or triple bonds, i.e., contain all single bonds.

The suffix -ene is used to further designate alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl groups as divalent bridging groups, such as alkylene, alkenylene, alkynylene, carbocyclylene, heterocyclylene, arylene, and heteroarylene.

Unless otherwise specifically provided, atoms, moieties or groups described herein may be unsubstituted or substituted as long as valence permits. The term "optionally substituted" means substituted or unsubstituted.

Unless otherwise expressly provided, group is optionally substituted.Term " optionally substituted " refers to be substituted or unsubstituted.In certain embodiments, alkyl, alkenyl, alkynyl, carbocyclic radical, heterocyclic radical, aryl and heteroaryl are optionally substituted (such as " substituted " or " unsubstituted " alkyl, " substituted " or " unsubstituted " alkenyl, " substituted " or " unsubstituted " alkynyl, " substituted " or " unsubstituted " carbocyclic radical, " substituted " or " unsubstituted " heterocyclic radical, " substituted " or " unsubstituted " aryl, or " substituted " or " unsubstituted " heteroaryl). Usually, term " substituted ", no matter whether or not there is term " optionally " before, mean that at least one hydrogen present in group (such as, carbon or nitrogen atom) is replaced by permissible substituent, such as substituent can form stable compound after replacement, such as, can not (such as by rearrangement, cyclization, elimination or other reactions) spontaneously transform compound. Unless otherwise indicated, a "substituted" group has a substituent at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituents are the same or different at each position. It is intended that the term "substituted" includes all permissible substituents of an organic compound, any of which are substituted with the substituents described herein that result in the formation of a stable compound. The present disclosure contemplates any and all of these combinations to obtain a stable compound. For the purposes of this disclosure, heteroatoms such as nitrogen may have hydrogen substituents and/or any suitable substituents that satisfy the valence of the heteroatoms as described herein and result in the formation of a stable moiety. In certain embodiments, the substituents are carbon atom substituents. In certain embodiments, the substituents are nitrogen atom substituents. In certain embodiments, the substituents are oxygen atom substituents. In certain embodiments, the substituents are sulfur atom substituents.

Exemplary carbon atom substituents include, but are not limited to, halogen, —CN, —NO ₂ , —N ₃ , —SO ₂ H, —SO ₃ H, —OH, —OR ^aa , —ON(R ^bb ) ₂ , —N(R ^bb ) ₂ , —N(R ^bb ) ₃ ⁺ X ⁻ , —N(OR ^cc )R ^bb , —SH, —SR ^aa , —SSR ^cc , —C(═O)R ^aa , —CO ₂ H, —CHO, —C(OR ^cc ) ₂ , —CO ₂ R ^aa , —OC(═O)R ^aa , —OCO ₂ R ^aa , —C(═O)N(R ^bb ) ₂ , —OC(═O)N(R ^bb ) ₂ , —NR ^bb C(═O)R ^aa , —NR ^bb CO ₂ R ^aa , —NR ^bb C(＝O)N(R ^bb ) ₂ , –C(＝NR ^bb )R ^aa , –C(＝NR ^bb )OR ^aa , –OC(＝NR ^bb )R ^aa , –OC(＝NR ^bb )OR ^aa , –C(＝NR ^bb )N(R ^bb ) ₂ , –OC(＝NR ^bb )N(R ^bb ) ₂ , –NR ^bb C(＝NR ^bb )N(R ^bb ) ₂ , –C(＝O)NR ^bb SO ₂ R ^aa , –NR ^bb SO ₂ R ^aa , –SO ₂ N(R ^bb ) ₂ , –SO ₂ R ^aa , –SO ₂ OR ^aa , –OSO ₂ R ^aa , –S(＝O)R ^aa , –OS(＝O)R ^aa , –Si(R ^aa ) ₃ , –OSi(R ^aa ) ₃ –C(=S)N(R ^bb ) ₂ , –C(=O)SR ^aa , –C(=S)SR ^aa , –SC(=S)SR ^aa , –SC(=O)SR ^aa , –OC(=O)SR ^aa , –SC(=O)OR ^aa , –SC(=O)R ^aa , –P(=O)(R ^aa ) ₂ , -P(＝O)(OR ^cc ) ₂ , –OP(＝O)(R ^aa ) ₂ , –OP(＝O)(OR ^cc ) ₂ , –P(＝O)(N(R ^bb )- ₂ ) ₂ , –OP(＝O)(N(R ^bb ) ₂ ) ₂ , –NR ^bb P(＝O)(R ^aa ) ₂ , –NR ^bb P(＝O)(OR ^cc ) ₂ , –NR ^bb P(＝O)(N(R ^bb ) ₂ ) _{2 ,} –P(R ^cc ) ₂ , -P _{( OR} ^cc ) ₂ ^, –P( ^{R cc )} ₃ ^{+ X –} _, ^{–P ( OR cc )} ₃ ^{+ , -OP (} _OR ^cc _{) 2 ,} ^{-OP (} _OR ^{cc )} ₃ ^{+ ​} _{​ ​ ​} C _2-10 alkynyl, C _3-10 carbocyclyl, 3-14 membered heterocyclyl, C _6-14 aryl and 5-14 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 R ^dd groups; wherein X ^- is a counterion;

or two geminal hydrogen groups on a carbon atom are replaced by groups =0, =S, =NN( ^Rbb ) ₂ , =NNRbbC(=O)Raa, = ^NNRbbC (=O) ^ORaa , = ^NNRbbS (=O) _2Raa , ^{= NRbb} or ^{= NORcc ;}

each instance of ^Raa is independently selected from Ci _-io alkyl, Ci _-io perhaloalkyl, C2 _-io alkenyl, C2 _-io alkynyl, C3 _-io carbocyclyl, 3-14 membered heterocyclyl, _C6-14 aryl, and 5-14 membered heteroaryl, or two ^Ra groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl group is independently substituted with 0, 1, 2, 3, 4, or 5 ^Rdd groups;

Each instance of R ^bb is independently selected from hydrogen, —OH, —OR ^aa , —N(R ^cc ) ₂ , —CN, —C(═O)R ^aa , —C(═O)N(R ^cc ) ₂ , —CO ₂ R ^aa , —SO ₂ R ^aa , —C(═NR ^cc )OR ^aa , —C(═NR ^cc )N(R ^cc ) ₂ , —SO ₂ N(R ^cc ) ₂ , —SO ₂ R ^cc , —SO ₂ OR ^cc , —SOR ^aa , —C(═S)N(R ^cc ) ₂ , —C(═O)SR ^cc , —C(═S)SR ^cc , —P(═O)(R ^aa ) ₂ , —P(═O)(OR ^cc ) ₂ , —P(═O)(N(R ^cc ) _{2 )} , C _{R 1-10} alkyl, C _1-10 perhaloalkyl, C _2-10 alkenyl, C _2-10 alkynyl, C _3-10 carbocyclyl, 3-14 membered heterocyclyl, C _6-14 aryl and 5-14 membered heteroaryl, or two R ^bb groups are linked to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 R ^dd groups; wherein X ^- is a counterion;

each instance of ^Rcc is independently selected from hydrogen, _C1-10 alkyl, _C1-10 perhaloalkyl, _C2-10 alkenyl, _C2-10 alkynyl, C3-10 carbocyclyl, _3-14 membered heterocyclyl, C6-14 aryl, and _5-14 membered heteroaryl, or two ^Rcc groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 ^Rdd groups;

Each instance of R ^dd is independently selected from halogen, —CN, —NO ₂ , —N ₃ , —SO ₂ H, —SO ₃ H, —OH, —OR ^ee , —ON(R ^ff ) ₂ , —N(R ^ff ) ₂ , —N(R ^ff ) ₃ ⁺ X ^– , —N(OR ^ee )R ^ff , —SH, —SR ^ee , —SSR ^ee , —C(═O)R ^ee , —CO ₂ H, —CO ₂ R ^ee , —OC(═O)R ^ee , —OCO ₂ R ^ee , —C(═O)N(R ^ff ) ₂ , —OC(═O)N(R ^ff ) ₂ , —NR ^ff C(═O)R ^ee , —NR ^ff CO ₂ R ^ee , —NR ^ff C(═O)N(R ^ff ) ₂ , –C(＝NR ^ff )OR ^ee , –OC(＝NR ^ff )R ^ee , –OC(＝NR ^ff )OR ^ee , –C(＝NR ^ff )N(R ^ff ) ₂ , –OC(＝NR ^ff )N(R ^ff ) ₂ , –NR ^ff C(＝NR ^ff )N(R ^ff ) ₂ , –NR ^ff SO ₂ R ^ee , –SO ₂ N(R ^ff ) ₂ , –SO ₂ R ^ee , –SO ₂ OR ^ee , –OSO ₂ R ^ee , –S(＝O)R ^ee , –Si(R ^ee ) ₃ , –OSi(R ^ee ) ₃ , –C(＝S)N(R ^ff ) ₂ , –C(＝O)SR ^ee , –C(＝S)SR ^ee , –SC(＝S)SR ^ee , –P(＝O)(OR ^ee ) ₂ , –P(＝O)(R ^ee ) ₂ , –OP(＝O)(R ^ee ) ₂ , –OP(＝O)(OR ^ee ) ₂ , C _1-6 alkyl, C _1-6 perhaloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-10 carbocyclyl, 3-10 membered heterocyclyl, C _6-10 aryl, and 5-10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R ^gg groups; or two geminal R ^dd substituents may be linked to form ＝O or ＝S; wherein X ^- is a counterion;

each instance of R ^ee is independently selected from C _1-6 alkyl, C _1-6 perhaloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-10 carbocyclyl, C _6-10 aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R ^gg groups;

each instance of ^R is independently selected from hydrogen, C _1-6 alkyl, C _1-6 perhaloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-10 carbocyclyl, 3-10 membered heterocyclyl, C _6-10 aryl, and 5-10 membered heteroaryl, or two ^R groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R ^gg groups; and

Each instance of R ^gg is independently halogen, —CN, —NO ₂ , —N ₃ , —SO ₂ H, —SO ₃ H, —OH, —OC _1-6 alkyl, —ON(C _1-6 alkyl) ₂ , —N(C _1-6 alkyl) ₂ , —N(C _1-6 alkyl) ₃ ⁺ X ^– , —NH(C _1-6 alkyl) ₂ ⁺ X ^– , —NH ₂ (C _1-6 alkyl) ⁺ X ^– , —NH ₃ ⁺ X ^– , —N(OC _1-6 alkyl)(C _1-6 alkyl), —N(OH)(C _1-6 alkyl), —NH(OH), —SH, —SC _1-6 alkyl, —SS(C _1-6 alkyl), —C(═O)(C _1-6 alkyl), —CO ₂ H, —CO ₂ (C _1-6 alkyl), —OC(═O)(C _1-6 alkyl), —OCO ₂ (C _1-6 alkyl), –C(＝O)NH ₂ , –C(＝O)N(C _1-6 alkyl) ₂ , –OC(＝O)NH(C _1-6 alkyl), –NHC(＝O)(C _1-6 alkyl), –N(C _1-6 alkyl)C(＝O)(C _1-6 alkyl), –NHCO ₂ (C 1-6 alkyl), –NHC(＝O)N(C _1-6 alkyl) ₂ , –NHC(＝O)NH(C _1-6 alkyl), –NHC(＝O)NH(C _1-6 alkyl), –NHC(＝O)NH ₂ , –C(＝NH)O(C _1-6 alkyl), –OC(＝NH)(C _1-6 alkyl), –OC(＝NH)OC _1-6 alkyl, –C(＝NH)N(C _1-6 alkyl) ₂ , –C(＝NH)NH(C _1-6 alkyl), –C(＝NH)NH ₂ , –OC(＝NH)N(C _1-6 alkyl) ₂ 、–OC(NH)NH(C _1-6 alkyl), –OC(NH)NH ₂ 、–NHC(NH)N(C _1-6 alkyl) ₂ 、–NHC(＝NH)NH ₂ 、–NHSO ₂ (C _1-6 alkyl), –SO ₂ N(C _1-6 alkyl) ₂ 、–SO ₂ NH(C _1-6 alkyl), –SO ₂ NH ₂ 、–SO ₂ C _1-6 alkyl, –SO ₂ OC _1-6 alkyl, –OSO ₂ C _1-6 alkyl, –SOC _1-6 alkyl, –Si(C _1-6 alkyl) ₃ 、–OSi(C _1-6 alkyl) ₃ 、–C(＝S)N(C _1-6 alkyl) ₂ 、C(＝S)NH(C _1-6 alkyl), C(＝S)NH ₂ 、–C(＝O)S(C _1-6 alkyl), –C(＝S)SC _1-6 alkyl, –SC(＝S)SC _1-6 alkyl, –P(＝O)(OC _1-6 alkyl) ₂ , –P(＝O)(C _1-6 alkyl) ₂ , –OP(＝O)(C _1-6 alkyl) ₂ , –OP(＝O)(OC _1-6 alkyl) ₂ , C _1-6 alkyl, C _1-6 perhaloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-10 carbocyclyl, C _6-10 aryl, 3-10 membered heterocyclyl and 5-10 membered heteroaryl; or two geminal R ^gg substituents may be linked to form ＝O or ＝S; wherein X ^- is a counterion;

Each instance of R ^gg is independently halogen, –CN, –NO ₂ , –N ₃ , –SO ₂ H, –SO ₃ H, –OH, –OC _1-6 alkyl, –ON(C _1-6 alkyl) ₂ , –N(C _1-6 alkyl) ₂ , –N(C _1-6 alkyl) ₃ ⁺ X ^– , –NH(C _1-6 alkyl) ₂ ⁺ X ^– , –NH ₂ (C _1-6 alkyl) ⁺ X ^– , –NH ₃ ⁺ X ^– , –N(OC _1-6 alkyl)(C _1-6 alkyl), –N(OH)(C _1-6 alkyl), –NH(OH), –SH, –SC _1-6 alkyl, –SS(C _1-6 alkyl), –C(═O)(C _1-6 alkyl), –CO ₂ H, –CO ₂ (C _1-6 alkyl), –OC(═O)(C _1-6 alkyl), –OCO ₂ (C _1-6 alkyl), –C(＝O)NH ₂ , –C(＝O)N(C _1-6 alkyl) ₂ , –OC(＝O)NH(C _1-6 alkyl), –NHC(＝O)(C _1-6 alkyl), –N(C _1-6 alkyl)C(＝O)(C _1-6 alkyl), –NHCO ₂ (C 1-6 alkyl), –NHC(＝O)N(C _1-6 alkyl) ₂ , –NHC(＝O)NH(C _1-6 alkyl), –NHC(＝O)NH(C _1-6 alkyl), –NHC(＝O)NH ₂ , –C(＝NH)O(C _1-6 alkyl), –OC(＝NH)(C _1-6 alkyl), –OC(＝NH)OC _1-6 alkyl, –C(＝NH)N(C _1-6 alkyl) ₂ , –C(＝NH)NH(C _1-6 alkyl), –C(＝NH)NH ₂ , –OC(＝NH)N(C _1-6 alkyl) ₂ 、–OC(NH)NH(C _1-6 alkyl), –OC(NH)NH ₂ 、–NHC(NH)N(C _1-6 alkyl) ₂ 、–NHC(＝NH)NH ₂ 、–NHSO ₂ (C _1-6 alkyl), –SO ₂ N(C _1-6 alkyl) ₂ 、–SO ₂ NH(C _1-6 alkyl), –SO ₂ NH ₂ 、–SO ₂ C _1-6 alkyl, –SO ₂ OC _1-6 alkyl, –OSO ₂ C _1-6 alkyl, –SOC _1-6 alkyl, –Si(C _1-6 alkyl) ₃ 、–OSi(C _1-6 alkyl) ₃ 、–C(＝S)N(C _1-6 alkyl) ₂ 、C(＝S)NH(C _1-6 alkyl), C(＝S)NH ₂ 、–C(＝O)S(C _1-6 alkyl), –C(＝S)SC _1-6 alkyl, –SC(＝S)SC _1-6 alkyl, –P(＝O)(OC _1-6 alkyl) ₂ , –P(＝O)(C _1-6 alkyl) ₂ , –OP(＝O)(C _1-6 alkyl) ₂ , –OP(＝O)(OC _1-6 alkyl) ₂ , C _1-6 alkyl, C _1-6 perhaloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-10 carbocyclyl, C _6-10 aryl, 3-10 membered heterocyclyl, 5-10 membered heteroaryl; or two geminal R ^gg substituents may be linked to form ＝O or ＝S; wherein X ^- is a counterion.

A "counter ion" or "anionic counter ion" is a negatively charged group associated with a positively charged group to maintain electronic neutrality. Anionic counter ions can be monovalent (i.e., include one formal negative charge). Anionic counter ions can also be multivalent (i.e., include more than one formal negative charge), such as divalent or trivalent. Exemplary counterions include halide ions (e.g., F ⁻ , Cl ⁻ , Br ⁻ , I ⁻ ), NO ₃ ⁻ , ClO ₄ ⁻ , OH ⁻ , H ₂ PO ₄ ⁻ , HSO ₄ ⁻ , sulfonate ions (e.g., methanesulfonate, trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate, 10-camphorsulfonate, naphthalene-2-sulfonate, naphthalene-1-sulfonic acid-5-sulfonate, ethane-1-sulfonic acid-2-sulfonate, etc.), carboxylate ions (e.g., acetate, propionate, benzoate, glycerate, lactate, tartrate, glycolate, gluconate, etc.), BF ₄ ⁻ , PF ₄ ⁻ , PF ₆ ⁻ , AsF ₆ ⁻ , SbF ₆ ⁻ , B[3,5-(CF ₃ ) ₂ C ₆ H ₃ ] ₄ ] ⁻ , BPh ₄ ⁻ , Al(OC(CF ₃ ) ₃ ) ₄ ⁻ , and carborane anions (e.g., CB ₁₁ H ₁₂ ⁻ or (HCB ₁₁ Me ₅ Br ₆ ) ⁻ ). Exemplary counterions that may be multivalent include CO ₃ ²⁻ , HPO ₄ ²⁻ , PO ₄ ³⁻ , B ₄ O ₇ ²⁻ , SO ₄ ²⁻ , S ₂ O ₃ ²⁻ , carboxylate anions (e.g., tartrate, citrate, fumarate, maleate, malate, malonate, gluconate, succinate, glutarate, adipate, pimelate, suberate, azelaate, sebacate, salicylate, phthalate, aspartate, glutamate, etc.), and carboranes.

"Halo" or "halogen" refers to fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br), or iodine (iodo, -I).

“Acyl” refers to a moiety selected from —C(═O)R ^aa , —CHO, —CO ₂ R ^aa , —C(═O)N(R ^bb ) ₂ , —C(═NR ^bb )R ^aa , —C(═NR ^bb )OR ^aa , —C(═NR ^bb )N(R ^bb ) ₂ , —C(═O)NR ^bb SO ₂ R ^aa , —C(═S)N(R ^bb ) ₂ , —C(═O)SR ^aa , or —C(═S)SR ^aa , where ^Ra and R ^bb are as defined herein.

Nitrogen atoms may be substituted or unsubstituted, as valency permits, and include primary, secondary, tertiary, and quaternary nitrogen atoms. Exemplary nitrogen atom substituents include, but are not limited to, hydrogen, —OH, —OR ^aa , —N(R ^cc ) ₂ , —CN, —C(═O)R ^aa , —C(═O)N(R ^cc ) ₂ , —CO ₂ R ^aa , —SO ₂ R ^aa , —C(═NR ^bb )R ^aa , —C(═NR ^cc )OR ^aa , —C(═NR ^cc )N(R ^cc ) ₂ , —SO ₂ N(R ^cc ) ₂ , —SO ₂ R ^cc , —SO ₂ OR ^cc , —SOR ^aa , —C(═S)N(R ^cc ) ₂ , —C(═O)SR ^cc , —C(═S)SR ^cc , —P(═O)(OR ^cc ) ₂ , —P(═O)(R ^aa ) ₂ , —P(═O)(N(R cc ) ^{R cc} ) ₂ ) ₂ , C _1-10 alkyl, C _1-10 perhaloalkyl, C _2-10 alkenyl, C _2-10 alkynyl, C _3-10 carbocyclyl, 3-14 membered heterocyclyl, C _6-14 aryl and 5-14 membered heteroaryl, or two R ^cc groups attached to a nitrogen atom are linked to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 R ^dd groups, and wherein ^Raa , R ^bb , R ^cc and R ^dd are as defined above.

In certain embodiments, a substituent present on a nitrogen atom is a nitrogen protecting group (also known as an amino protecting group). Nitrogen protecting groups include, but are not limited to, –OH, –OR ^aa , –N(R ^cc ) ₂ , –C(═O)R ^aa , –C(═O)N(R ^cc ) ₂ , –CO ₂ R ^aa , –SO ₂ R ^aa , –C(═NR ^cc )R ^aa , –C(═NR ^cc )OR ^aa , –C(═NR ^cc )N(R ^cc ) ₂ , –SO ₂ N(R ^cc ) ₂ , –SO ₂ R ^cc , –SO ₂ OR ^cc , –SOR ^aa , –C(═S)N(R ^cc ) ₂ , –C(═O)SR ^cc , –C(═S)SR ^cc , C _1-10 alkyl (e.g., aralkyl, heteroaralkyl), C _2-10 alkenyl, C _2-10 alkynyl, , _C6-14 aryl, and 5-14 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aralkyl, aryl, and heteroaryl is independently substituted with ₀ , 1, 2, 3, 4, or 5 ^Rdd groups, and wherein ^Raa , ^Rbb , ^Rcc , and ^Rdd are as defined herein. Nitrogen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, ^3rd edition, John Wiley & Sons, 1999, which is incorporated herein by reference.

For example, nitrogen protecting groups, such as amides (e.g., —C(═O) ^Raa ), include, but are not limited to, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropionamide, picolinamide, 3-pyridineamide, N-benzoylphenylalanyl derivatives, benzamide, p-phenylbenzamide, o-nitrophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide, (N′-dithiobenzyloxyamido)acetamide, 3-(p-hydroxyphenyl)propionamide, 3-(o-nitrophenyl)propionamide, 2-methyl-2-(o-nitrophenoxy)propionamide, 2-methyl-2-(o-phenylazophenoxy)propionamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinnamic acid, N-acetylmethionine derivatives, o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.

Nitrogen protecting groups, such as carbamate groups (e.g., -C(=O)OR ^aa ), include, but are not limited to, methyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-thio)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluorenylmethyl carbamate, 2,7-di-tert-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthenyl)]methylcarbamate (DBD-Tmoc), 4-methoxybenzoylmethyl carbamate (Phenoc), 2,2,2-trichloroethyl carbamate (Troc), carbamate 2-trimethylsilylethyl carbamate (Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethyl carbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC), 1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc), 1-(3,5-dichloroethyl carbamate -tert-butylphenyl)-1-methylethyl ester (t-Bumeoc), 2-(2'- and 4'-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylamide)ethyl carbamate, tert-butyl carbamate (BOC or Boc), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate Ester (Noc), 8-quinolinyl carbamate, N-hydroxypiperidinyl carbamate, alkyl dithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p-nitrobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate, 2-(p-Toluenesulfonyl)ethyl ester, [2-(1,3-dithiocyclohexyl)]methyl carbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc), 2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphoniumethyl carbamate (Peoc), 2-triphenylphosphoniumisopropyl carbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl carbamate, 5-benzisothiazolyl carbamate Oxazolyl methyl ester, 2-(trifluoromethyl)-6-chromonyl methyl carbamate (Tcroc), m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl (o-nitrophenyl) methyl carbamate, tert-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentyl carbamate, cyclopropyl methyl carbamate, p-decyloxybenzyl carbamate, amino 2,2-dimethoxyacylvinyl carbamate, o-(N,N-dimethylamido)benzyl carbamate, 1,1-dimethyl-3-(N,N-dimethylamido)propyl carbamate, 1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate, 2-furylmethyl carbamate, 2-iodoethyl carbamate, isobornyl carbamate, isobutyl carbamate, isonicotinoyl carbamate, p-(p-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcarbamate carbamate, 1-methyl-1-cyclopropylmethyl carbamate, 1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate, 1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate, 2,4,6-tri-tert-butylphenyl carbamate, 4-(trimethylammonium)benzyl carbamate and 2,4,6-trimethylbenzyl carbamate.

Nitrogen protecting groups, for example, sulfonamide groups (eg, -S(=O) ₂ R ^aa ) include, but are not limited to, p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide, 4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide, and phenacylmethylsulfonamide.

Other nitrogen protecting groups include, but are not limited to, phenothiazinyl-(10)-acyl derivatives, N'-toluenesulfonylaminoacyl derivatives, N'-phenylaminothioacyl derivatives, N-benzoylphenylalanyl derivatives, N-acetylmethionine derivatives, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilazide adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one , 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl)amine, quaternary ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenamine (PhF), N-2, 7-Dichloro-9-fluorenylmethylamine, N-ferrocenylmethylamino (Fcm), N-2-picolylamino N'-oxide, N-1,1-dimethylsulfomethyleneamine, N-benzylideneamine, N-p-methoxybenzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)trimethylphenyl]methyleneamine, N-(N',N'-dimethylaminomethylene)amine, N,N'-isopropylenediamine, N-p-nitrobenzylideneamine, N-salicylideneamine, N-5-chlorosalicylideneamine, N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexyleneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl) Amines, N-borane derivatives, N-diphenylboronic acid derivatives, N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper chelates, N-zinc chelates, N-nitramines, N-nitrosamines, amine N-oxides, diphenylphosphinamide (Dpp), dimethylphosphinamide (Mpt), diphenylphosphinamide (Ppt), dialkylphosphoramidates, dibenzylphosphoramidates, diphenylphosphoramidates, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, and 3-nitropyridinesulfenamide (Npys).

Exemplary oxygen atom substituents include, but are not limited to, —R ^aa , —C(═O)SR ^aa , —C(═O)R ^aa , —CO ₂ R ^aa , —C(═O)N(R ^bb ) ₂ , —C(═NR ^bb )R ^aa , —C(═NR ^bb )OR ^aa , —C(═NR ^bb )N(R ^bb ) ₂ , —S(═O)R ^aa , —SO ₂ R ^aa , —Si(R ^aa ) ₃ , —P(R ^cc ) ₂ , —P(R ^cc ) ₃ ⁺ X ⁻ , —P(OR ^cc ) ₂ , —P(OR ^cc ) ₃ ⁺ X ⁻ , —P(═O)(R ^aa ) ₂ , —P(═O)(OR ^cc ) ₂ , and —P(═O)(N(R ^bb ) ₂ ) _{2 .} , wherein ^X- , ^Raa , ^Rbb and ^Rcc are as defined herein. In certain embodiments, the oxygen atom substituent present on the oxygen atom is an oxygen protecting group (also referred to as a hydroxy protecting group). Oxygen protecting groups are those well known in the art and are included in those described in detail in "Protecting Groups in Organic Synthesis", TW Greene and PGM Wuts, ^3rd edition, John Wiley & Sons, 1999, which are incorporated herein by reference. Exemplary oxygen protecting groups include, but are not limited to, methyl, tert-butyloxycarbonyl (Boc or Boc), methoxymethyl (MOM), methylthiomethyl (MTM), tert-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacylmethyl (GUM), tert-butoxymethyl, 4-pentenyloxymethyl (POM), silyloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMO), R), tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl (CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanolbenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl 1-Benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylhydrogen selenyl)ethyl, tert-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxyl, diphenylmethyl, p,p'-dinitrodiphenylmethyl, 5-dibenzosuberyl, triphenylmethyl, α-naphthyldiphenylmethyl, p-methoxyphenyldiphenyl methyl, bis(p-methoxyphenyl)phenylmethyl, tris(p-methoxyphenyl)methyl, 4-(4'-bromophenacylmethoxyphenyl)diphenylmethyl, 4,4',4"-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4',4"-tris(levulinyloxyphenyl)methyl, 4,4',4"-tris(benzoyloxyphenyl)methyl, 3-(imidazol-1-yl)bis(4',4"-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxy)anthryl, 1,3-benzodithiolan-2-yl (1,3-benzodisol-2-yl) ulfuran-2-yl), S,S-dioxybenzisothiazolyl, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethylhexylsilyl (dimethylthexylsilyl), tert-butyldimethylsilyl (TBDMS), tert-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), tert-butylmethoxyphenylsilyl (TBDPS), MPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (levulinic acid ester), 4,4-(ethylenedithio)pentanoate (levulinic acid dithioacetal), pivaloate, adamantanoate (adamantoate), crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc) , alkyl ethyl carbonate, alkyl 2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec), 2-(triphenylphosphonium)ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl ethylene carbonate, alkyl allyl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro -4-Methylvalerate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinate, (E)-2-methyl-2-butenoate, o-(methoxyacyl)benzoate, α-naphthoate, nitrate, N,N,N',N'-tetramethylphosphorodiamidate alkyl esters (alkyl N,N,N',N'-tetramethylphosphorodiamidate), alkyl N-phenylcarbamate, borate, dimethylphosphinothioate, alkyl 2,4-dinitrophenylsulfinate, sulfate, mesylate, benzylsulfonate, and tosylate (Ts).

"Hydrocarbon chain" refers to a substituted or unsubstituted divalent alkyl, alkenyl or alkynyl group. A hydrocarbon chain includes (1) one or more carbon atom chains immediately between two groups of the hydrocarbon chain; (2) one or more hydrogen atoms optionally on the carbon atom chain; and (3) one or more substituents ("non-chain substituents" which are not hydrogen) optionally on the carbon atom chain. A carbon atom chain consists of continuously linked carbon atoms ("chain atoms" or "carbon units") and does not include hydrogen atoms or heteroatoms. However, a non-chain substituent of a hydrocarbon chain may include any atom, including hydrogen atoms, carbon atoms and _heteroatoms . For example, the _hydrocarbon chain ^-CAH ( ^CBH2CCH3 )- includes one chain atom ^CA , one hydrogen atom on ^CA and a non-chain substituent -( ^{CBH2CCH3 )} . The term " _Cx hydrocarbon chain", where x is a positive integer, refers to a hydrocarbon chain including x chain atoms between _the two groups of the hydrocarbon chain ^. If there is more _than one possible value for x, the smallest possible value of x is used to define the hydrocarbon chain. For example, -CH( _C2H5 )- is a _C1 hydrocarbon chain, while -CH( _C2H5 )- is a _C3 hydrocarbon chain. When a range of values is used, the meaning of the range is as described herein. For example, a _C3-10 hydrocarbon chain refers to a hydrocarbon chain in which the number of chain atoms of the shortest chain of carbon atoms immediately adjacent to two groups of the hydrocarbon chain is 3, 4, 5, 6, 7, 8, 9, or 10. The hydrocarbon chain can be saturated (e.g., -( _CH2 ) _4- ). The hydrocarbon chain can also be unsaturated and include one or more C=C and/or C≡C bonds anywhere in the hydrocarbon chain. For example, -CH=CH-( _CH2 ) _2- , _-CH2 -C≡C- _CH2- , and -C≡C-CH=CH- are all examples of unsubstituted and unsaturated hydrocarbon chains. In certain embodiments, the hydrocarbon chain is unsubstituted (e.g., -C≡C- or -( _CH2 ) _4- ). In certain embodiments, the hydrocarbon chain is substituted (e.g., -CH( _C2H5 )- and _-CF2- ). Any two substituents on _the hydrocarbon chain can be linked to form an optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl ring. For example, are all examples of hydrocarbon chains. In contrast, in certain embodiments, are not within the scope of the hydrocarbon chains described herein. When a chain atom of a _Cx hydrocarbon chain is replaced by a heteroatom, the resulting group is referred to as a _Cx hydrocarbon chain in which the chain atom is replaced by a heteroatom, rather than a Cx _-1 hydrocarbon chain. For example, is a _C3 hydrocarbon chain in which one of the chain atoms is replaced by an oxygen atom.

The term "leaving group" has the common meaning in the field of synthetic organic chemistry, and refers to an atom or group that can be replaced by a nucleophilic reagent. For example, see Smith, "March Advanced Organic Chemistry", 6th edition, (501-502) (Smith, March Advanced Organic Chemistry ^6th ed. (501-502)). The example of a suitable leaving group includes but is not limited to halogen (such as F, Cl, Br or I (iodine)), alkoxycarbonyloxy, aryloxycarbonyloxy, alkanesulfonyloxy, arylsulfonyloxy (arenesulfonyloxy), alkylcarbonyloxy (such as acetoxy), arylcarbonyloxy, aryloxy, methoxyl group, N, O-dimethylhydroxylamino, 9-phenylxanthene (pixyl) and haloformate. In some cases, the leaving group is an activated substituted hydroxyl group (e.g., —OC(═O)SR ^aa , —OC(═O)R ^aa , —OCO ₂ R ^aa , —OC(═O)N(R ^bb ) ₂ , —OC(═NR ^bb )R ^aa , —OC(═NR ^bb )OR ^aa , —OC(═NR ^bb )N(R ^bb ) ₂ , —OS(═O)R ^aa , —OSO ₂ R ^aa , —OP(R ^cc ) ₂ , —OP(R ^cc ) ₃ , —OP(═O) ₂ R ^aa , —OP(═O)(R ^aa ) ₂ , —OP(═O)(OR ^cc ) ₂ , —OP(═O) ₂ N(R ^bb ) ₂ , and —OP(═O)(NR ^bb ) ₂ ), wherein ^Raa , R ^bb , and R ^cc is as defined herein). In some cases, the leaving group is a sulfonate, such as tosylate (-OTs), mesylate (-OMs), bromobenzenesulfonyloxy (brosylate, -OBs), -OS(=O) ₂ (CF ₂ ) ₃ CF ₃ (nonaflate, -ONf), or triflate (triflate, -OTf). In some cases, the leaving group is a bromobenzenesulfonate, such as bromobenzenesulfonyloxy. In some cases, the leaving group is a nitrobenzenesulfonate (nosylate), such as 2-nitrobenzenesulfonyloxy. In some embodiments, the leaving group is a sulfonate-containing group. In some embodiments, the leaving group is a tosylate. The leaving group can also be a phosphine oxide (e.g., formed during a Mitsunobu reaction) or an internal leaving group, such as an epoxide or a cyclic sulfate. Other non-limiting examples of leaving groups are water, ammonia, alcohols, ether moieties, thioether moieties, zinc halides, magnesium moieties, diazonium salts, and copper moieties.

The term "pharmaceutically acceptable salt" refers to salts that are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, etc., and that are commensurate with a reasonable benefit/risk ratio, within the scope of sound medical judgment. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, which are incorporated herein by reference. Pharmaceutically acceptable salts of the compounds described herein include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are amino salts formed with inorganic acids (such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid) or with organic acids (acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid) or by using other methods known in the art (such as ion exchange). Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydrogen iodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, dodecylsulfate, 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, p-toluenesulfonate, undecanoate, valerate, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ ^-salts . Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium and the like. Other pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates and aryl sulfonates, where appropriate.

The term "solvate" refers to a compound form that is typically bound to a solvent by a solvolysis reaction. This physical binding can include hydrogen bonds. Conventional solvents include water, methanol, ethanol, acetic acid, DMSO, THF, diethyl ether, and the like. The compounds described herein can be prepared, for example, in crystalline form and can be solvated. Suitable solvates include pharmaceutically acceptable solvates, and also include stoichiometric solvates and non-stoichiometric solvates. In some cases, for example, when one or more solvent molecules are incorporated into the crystal lattice of the crystalline solid, the solvate will be able to separate. "Solvate" includes solution phase and separable solvates. Representative solvates include hydrates, ethanolates, and methoxides.

The term "hydrate" refers to a compound that is combined with water. Generally, the number of water molecules contained in a hydrate of a compound is proportional to the number of molecules of the compound in the hydrate. Thus, a hydrate of a compound can, for example, be represented by the general formula R·xH ₂ O, where R is a compound and x is a number greater than 0. A given compound can form more than one type of hydrate, including, for example, monohydrates (x is 1), lower hydrates (x is a number greater than 0 and less than 1, such as hemihydrate (R·0.5H ₂ O)), and polyhydrates (x is a number greater than 1, such as dihydrate (R·2H ₂ O) and hexahydrate (R·6H ₂ O)).

The term "tautomer" or "tautomeric" refers to two or more interconvertible compounds resulting from at least one formal migration of a hydrogen atom and at least one change in valence (e.g., a single bond becomes a double bond, a triple bond becomes a single bond, and vice versa). The exact ratio of tautomers depends on several factors, including temperature, solvent, and pH. Tautomerization reactions (i.e., reactions that provide tautomeric pairs) can be catalyzed by acids or bases. Exemplary tautomerization reactions include keto-enol, amide-imide, lactam-lactim, enamine-imine, and enamine-(different enamine) tautomerization reactions.

It is also understood that compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed "isomers." Isomers that differ in the arrangement of their atoms in space are termed "stereoisomers."

Stereoisomers that are not mirror images of each other are termed "diastereomers," and stereoisomers that are non-superimposable mirror images of each other are termed "enantiomers." When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers may exist. Enantiomers can be characterized by the absolute configuration of their asymmetric center and described by Cahn and Prelog's rules of R- and S-sequencing or by the way the molecule rotates the plane of polarized light, and are denoted as right- or left-handed (i.e., (+) or (-)-isomers, respectively). Chiral compounds can exist as individual enantiomers or as mixtures thereof. A mixture containing equal proportions of enantiomers is termed a "racemic mixture."

The term "polymorph" refers to a crystalline form of a compound (or its salt, hydrate or solvate) arranged in a specific crystal packing. All polymorphs have the same elemental composition. Different crystalline forms typically have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardnesses, crystal shapes, optical and electrical properties, stability and solubility. Recrystallization solvents, crystallization rates, storage temperatures and other factors may lead to a dominant crystalline form. Various polymorphs of a compound can be prepared by crystallization under different conditions.

The term "prodrug" refers to a compound having a cleavable group and being converted into a compound described herein by solvolysis or under physiological conditions, which has pharmaceutical activity in vivo. Such examples include, but are not limited to, choline ester derivatives, N-alkylmorpholine esters, and the like. Other derivatives of the compounds described herein, both in their acid and acid derivative forms, are active, but acid-sensitive forms generally provide advantages of solubility, tissue compatibility, or delayed release in mammalian organisms (see Bundgard, H., Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs include acid derivatives well known to those skilled in the art, for example, esters prepared by reacting the parent acid with a suitable alcohol, or amides prepared by reacting the parent acid compound with a substituted or unsubstituted amine, or anhydrides or mixed anhydrides. Simple aliphatic or aromatic esters, amides, and anhydrides derived from pendant acidic groups of the compounds described herein are specific prodrugs. In some cases, it is desirable to prepare double ester type prodrugs, such as (acyloxy)alkyl esters or ((alkoxycarbonyl)oxy)alkyl esters. Preferred may be C ₁ -C ₈ alkyl esters, C 2 -C ₈ alkenyl esters, C _{2 -C 8} alkynyl esters, aryl esters, C ₇ -C ₁₂ substituted aryl esters, and C ₇ -C ₁₂ arylalkyl esters of the compounds described herein.

The term "inhibit" or "inhibitor" refers to the ability of a compound to reduce, slow, arrest, or prevent the activity of a particular biological process (eg, the activity of an IDO enzyme relative to a vector in a cell).

When a compound, pharmaceutical composition, method, use, or kit is referred to as "selectively," "specifically," or "competitively" inhibiting the activity of an IDO enzyme, the compound, pharmaceutical composition, method, use, or kit inhibits the activity of the IDO enzyme to a greater extent (e.g., not less than about 2-fold, not less than about 5-fold, not less than about 10-fold, not less than about 30-fold, not less than about 100-fold, not less than about 1,000-fold, or not less than about 10,000-fold) as compared to the activity of at least one protein other than the IDO enzyme.

The term "abnormal activity" refers to an activity that deviates from normal activity. The term "increased activity" refers to an activity that is higher than normal activity.

The terms "composition" and "formulation" are used interchangeably.

A "subject" intended for administration is a human (i.e., male or female of any age group, such as a pediatric subject (e.g., infant, child, or adolescent) or an adult subject (e.g., young adult, middle-aged, or elderly)). A "patient" is a human subject in need of treatment for a disease.

The term "biological sample" refers to any sample including tissue samples (e.g., tissue sections and needle biopsies of tissue); cell samples (e.g., cytological smears (e.g., Pap smears or blood smears) or cell samples obtained by microdissection); samples of whole organisms (e.g., yeast or bacterial samples); or cell parts, fragments, or organelles (e.g., obtained by lysing cells and separating their components by centrifugation or other means). Other examples of biological samples include blood, serum, urine, semen, fecal matter, cerebrospinal fluid, interstitial fluid, mucus, tears, sweat, pus, biopsy tissue (e.g., obtained by surgical biopsy or needle biopsy), nipple aspirate, milk, vaginal fluid, saliva, swabs (e.g., buccal swabs), or any material containing biomolecules derived from a first biological sample.

The term "administering" refers to implanting, absorbing, ingesting, injecting, inhaling, or otherwise introducing a compound described herein, or a composition thereof, into or onto a subject.

The term "treat" refers to reversing, alleviating, delaying the onset of a disease described herein, or inhibiting the development of a disease described herein. In some embodiments, treatment may be administered after one or more signs or symptoms of the disease have developed or have been observed. In other embodiments, treatment may be administered in the absence of signs or symptoms of the disease. For example, treatment may be administered to a susceptible subject prior to the onset of symptoms (e.g., based on a history of symptoms and/or based on exposure to a pathogen) to delay or prevent the onset of the disease. Treatment may also be continued after symptoms subside, for example, to delay or prevent recurrence.

The terms "condition," "disease," and "disorder" are used interchangeably.

An "effective amount" of a compound described herein is an amount sufficient to elicit the desired biological response (i.e., to treat a condition). As will be appreciated by one of ordinary skill in the art, the effective amount of a compound described herein can vary depending on factors such as the desired biological endpoint, the pharmacokinetics of the compound, the condition being treated, the mode of administration, and the age and health of the subject. In certain embodiments, the effective amount is a therapeutically effective amount. In certain embodiments, the effective amount is a prophylactic treatment. In certain embodiments, the effective amount is the amount of a compound described herein in a single dose. In certain embodiments, the effective amount is the combined amount of the compounds described herein in multiple doses.

A "therapeutically effective amount" of a compound described herein is an amount sufficient to provide a therapeutic benefit in treating a condition or to delay or minimize one or more symptoms associated with the condition. A therapeutically effective amount of a compound refers to an amount of the therapeutic agent that, alone or in combination with other therapies, provides a therapeutic benefit in treating the condition. The term "therapeutically effective amount" can include an amount that improves overall treatment, reduces or avoids symptoms, signs, or causes, and/or enhances the therapeutic efficacy of another therapeutic agent.

A "prophylactically effective amount" of a compound described herein is an amount sufficient to prevent a condition, or one or more symptoms associated with the condition, or to prevent its recurrence. A prophylactically effective amount of a compound means an amount of a therapeutic agent that, alone or in combination with other agents, provides a prophylactic benefit in preventing the condition. The term "prophylactically effective amount" can include an amount that improves overall prevention or enhances the prophylactic efficacy of another prophylactic agent.

"Proliferative disease" refers to a disease that occurs due to abnormal growth of cells or spread due to proliferation (Walker, Cambridge Dictionary of Biology; Cambridge University Press: Cambridge, UK, 1990). Proliferative diseases may be associated with: 1) pathological proliferation of normally dormant cells; 2) pathological migration of cells from their normal location (e.g., tumor cell metastasis); 3) pathological expression of proteolytic enzymes, such as matrix metalloproteinases (e.g., collagenases, gelatinases, and elastases); or 4) pathological angiogenesis, as in proliferative retinopathy and tumor metastasis. Exemplary proliferative diseases include cancer (i.e., "malignancies"), benign tumors, angiogenesis, inflammatory diseases, and autoimmune diseases.

The terms "neoplasm" and "tumor" are used interchangeably herein and refer to an abnormal mass of tissue in which the growth of the mass exceeds that of normal tissue and is not coordinated with the growth of normal tissue. A tumor may be "benign" or "malignant" based on the following characteristics: degree of cell differentiation (including morphology and function), growth rate, local infiltration and metastasis. "Benign tumors" are usually well differentiated, have slower growth than malignant tumors, and are still confined to the site of origin. In addition, benign tumors do not have the ability to infiltrate, invade or metastasize to distant sites. Exemplary benign tumors include but are not limited to lipomas, chondromas, adenomas, warts, senile hemangiomas, seborrheic keratoses, freckles and sebaceous hyperplasia. In some cases, some "benign" tumors may later cause malignant tumors, which may be caused by other genetic changes in the tumor cell subpopulation of the tumor, and these tumors are referred to as "pre-malignant neoplasms". An exemplary pre-malignant neoplasm is a teratoma. In contrast, " malignant tumor " is poorly differentiated (degeneration) usually, and has the characteristic rapid growth that is accompanied by the progressive infiltration of surrounding tissue, invades and destroys.In addition, malignant tumor usually has the ability that is transferred to distant site.Term " transfer (metastasis) ", " transfer (metastatic) " or " transfer (metastasize) " refer to that cancer cell spreads or migrates to another organ or tissue from primary or original tumor, and can usually be identified by the existence of " secondary tumor " or " secondary cell mass " of the tissue type of primary or original tumor rather than the existence of the tissue type of the organ where secondary (metastatic) tumor is located or tissue.For example, the prostate cancer that has migrated to bone is considered as the prostate cancer of transfer, and is included in the cancerous prostate cancer cell that grows in bone tissue.

The term "cancer" refers to a class of diseases characterized by the development of abnormal cells that proliferate uncontrollably and have the ability to infiltrate and destroy normal body tissues. See, for example, Stedman's Medical Dictionary, 25th ed.; Hensyl ed.; Williams & Wilkins: Philadelphia, 1990. Exemplary cancers include, but are not limited to, hematological malignancies. Other exemplary cancers include, but are not limited to, acoustic neuroma; adenocarcinoma; adrenal cancer; anal cancer; angiosarcoma (e.g., lymphangiosarcoma, lymphoendothelioma, hemangioendothelioma); appendix cancer; benign monoclonal gammopathy; bile cancer (e.g., bile duct cancer); bladder cancer; breast cancer (e.g., adenocarcinoma of the breast, papillary carcinoma of the breast, breast cancer, medullary carcinoma of the breast); brain cancer (e.g., meningioma, glioblastoma, glioma (e.g., astrocytoma, oligodendroglioma), medulloblastoma); bronchogenic carcinoma; carcinoid tumor; cervical cancer (e.g., cervical adenocarcinoma); choriocarcinoma; chordoma; craniopharyngioma; colorectal cancer (e.g., colon cancer, rectal cancer, colorectal adenocarcinoma); connective tissue cancer; epithelial cancer; ependymoma; endothelial sarcoma (e.g., Kaposi's sarcoma); sarcoma), most idiopathic hemorrhagic sarcomas); endometrial cancer (e.g., uterine cancer, uterine sarcoma); esophageal cancer (e.g., esophageal adenocarcinoma, Barrett's adenocarcinoma); Ewing's sarcoma; eye cancer (e.g., intraocular melanoma, retinoblastoma); familial hypereosinophilia; gallbladder cancer; gastric cancer (e.g., gastric adenocarcinoma); gastrointestinal stromal tumor (GIST); germ cell cancer; head and neck cancer (e.g., head and neck squamous cell carcinoma, oral cancer (e.g., oral squamous cell carcinoma), pharyngeal cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer)); heavy chain disease (e.g., alpha chain disease, gamma chain disease, mu chain disease); hemangioblastoma; hypopharyngeal cancer; inflammatory myofibroblastic tumor; immune cell amyloidosis; renal cancer (e.g., renal embryonal tumor, also known as Wilms' tumor); renal cell carcinoma); liver cancer (e.g., hepatocellular carcinoma (HCC), malignant liver cancer); lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), lung adenocarcinoma); leiomyosarcoma (LMS); mastocytosis (e.g., systemic mastocytosis); muscle cancer; myelodysplastic syndrome (MDS); mesothelioma; myeloproliferative disorders (MPDs) (e.g., polycythemia vera (PV), essential thrombocythemia (ET), also known as myelofibrosis (MF) amyloid metaplasia (AMM), chronic idiopathic myelofibrosis, chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES)); neuroblastoma; neurofibromas (e.g., neurofibromatosis type 1 or type 2, schwannomatosis); neuroendocrine cancers (e.g., gastroenteropancreatic neuroendocrine tumors (GEP-NETs), carcinoid tumors); osteosarcomas (e.g., bone cancer); ovarian cancers (e.g., cystadenocarcinoma, embryonal carcinoma of the ovary). , ovarian adenocarcinoma); papillary adenocarcinoma; pancreatic cancer (e.g., pancreatic adenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), islet cell tumor); penile cancer (e.g., Paget's disease of the penis and scrotum); pinealoma; primitive neuroectodermal tumor (PNT); plasmacytoma; paraneoplastic syndrome; intraepithelial neoplasia; prostate cancer (e.g., prostate adenocarcinoma); rectal cancer; rhabdomyosarcoma; salivary gland cancer; skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)); small bowel cancer (e.g., bowel cancer (e.g., appendix cancer); soft tissue sarcomas (e.g., malignant fibrous histiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma); sebaceous gland cancer; small intestine cancer; sweat gland cancer; synovioma; testicular cancer (e.g., seminoma, testicular embryonal carcinoma); thyroid cancer (e.g., papillary thyroid carcinoma, papillary thyroid carcinoma (PTC), medullary thyroid cancer); urethral cancer; vaginal cancer; and vulvar cancer (e.g., Paget's disease of the vulva).

DETAILED DESCRIPTION

The present disclosure provides indoleamine 2,3-dioxygenase (IDO) inhibitors, such as compounds of formula (I). The compounds described herein can be used to treat and/or prevent proliferative diseases (such as cancer) by inhibiting IDO and inhibiting tryptophan catabolism, resulting in reduced kynurenine levels. The exemplary IDO inhibitory compounds described herein successfully demonstrated in vitro efficacy and in vivo efficacy. In addition, compared with other IDO inhibitors known in the art, such as INCB-24360 and other compounds disclosed in WO2014150677 and WO2014150646, these compounds showed better efficacy and/or lower human hepatocyte clearance. The present disclosure also provides pharmaceutical compositions, kits, and methods for treating proliferative diseases (such as cancer) using the IDO inhibitors described herein.

IDO inhibitory compounds

One aspect of the present disclosure relates to IDO inhibitory compounds as described herein and pharmaceutically acceptable salts, solvates, hydrates, polymorphs, cocrystals, tautomers, stereoisomers, isotopically labeled derivatives or prodrugs thereof. These compounds can be used to treat and/or prevent a proliferative disease in a subject.

In certain embodiments, the compound described herein is a compound of Formula (I) or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof:

wherein R ¹ -R ⁶ , W, Y and Q are as described herein.

Formula (I) includes a linker W connecting substituents R ¹ , R ² , and R ³ to the aromatic ring containing Y. In some embodiments, W can be -O-. In some embodiments, W can be -S-. In some embodiments, W can be a bond.

In addition, Formula (I) includes a linker Q connecting the substituent R ⁶ to the -NH- linker connected to the aromatic ring containing Y. In some embodiments, Q can be -C(=O)NH-. In some embodiments, Q can be a bond. In some embodiments, Q(R ⁶ ) can be

In formula (I), Y is in an aromatic ring. In some embodiments, Y is -CR ⁸ =, wherein R ⁸ is as defined therein. In some embodiments, R ⁸ can be hydrogen. In some embodiments, R ⁸ can be halogen (e.g., F, Cl, Br, or I). In some embodiments, R ⁸ can be -CN. In some embodiments, R ⁸ can be -OH. In some embodiments, R ⁸ can be substituted or unsubstituted C _1-6 alkyl (e.g., methyl, ethyl, propyl, or butyl). In some embodiments, R ⁸ can be substituted or unsubstituted C _1-6 alkoxy (e.g., substituted or unsubstituted methoxy or ethoxy). In one example, Y can be -CH=.

In some embodiments, ^R in formula (I) can be -C(=O)OH. In some embodiments, ^R can be a substituted or unsubstituted heterocyclyl (e.g., a substituted or unsubstituted 3 to 9-membered monocyclic heterocyclyl containing 0, 1 or 2 double bonds in the heterocyclic ring system, wherein 1, 2, 3 or 4 atoms in the heterocyclic ring system are independently nitrogen, oxygen or sulfur). In some embodiments, ^R can be a substituted or unsubstituted heteroaryl (e.g., a substituted or unsubstituted 5 to 6-membered monocyclic heteroaryl, wherein 1, 2, 3 or 4 atoms in the heteroaryl ring system are independently nitrogen, oxygen or sulfur). In some embodiments, ^R is a substituted or unsubstituted 5-membered heteroaryl. In certain embodiments, ^R is a substituted or unsubstituted 6-membered heteroaryl. In some embodiments, R ¹ can be of the formula: In some embodiments, R ¹ can be -NHSO ₂ R ⁹ or -C(=O)NHSO ₂ R ⁹ , wherein R ⁹ is as defined herein. In some embodiments, R ⁹ can be hydrogen. In some embodiments, R ⁹ can be substituted or unsubstituted C _1-6 alkyl (e.g., methyl, ethyl, propyl, or butyl). In some embodiments, R ⁹ can be substituted or unsubstituted C ₂ -C ₆ alkenyl.

In some embodiments, R ¹⁰ can be -C(=O)NHC(=O)OR ¹⁰ or -SO ₂ NHC(=O)R ¹⁰ , wherein R ¹⁰ is as defined herein. In some embodiments, R ¹⁰ can be hydrogen. In some embodiments, R ¹⁰ can be substituted or unsubstituted C _1-6 alkyl (e.g., methyl, ethyl, propyl, or butyl). In some embodiments, R ¹⁰ can be substituted or unsubstituted C ₂ -C ₆ alkenyl.

In some embodiments, R ¹ is -C(=O)OR ¹⁰ , wherein R ¹⁰ is as defined herein. For example, R ¹ can be optionally substituted -C(=O)OC _1-6 alkyl.

In some embodiments, R ² and/or R ³ in formula (I) can be hydrogen. In some embodiments, R ² and/or R ³ can be halogen (e.g., F, Cl, Br, or I). In some embodiments, R ² and/or R ³ can be substituted or unsubstituted C _1-6 alkyl (e.g., substituted or unsubstituted, methyl, ethyl, propyl, or butyl). In some embodiments, R ² and/or R ³ can be substituted or unsubstituted C _1-6 alkoxy (e.g., substituted or unsubstituted methoxy or ethoxy).

In some embodiments, R ² and R ³ can be connected to form a substituted or unsubstituted monocyclic or bicyclic 3 to 8-membered carbocyclic ring. In some embodiments, R ² and R ³ can be connected to form a substituted or unsubstituted monocyclic or bicyclic 3 to 6-membered carbocyclic ring (e.g., substituted or unsubstituted cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In some embodiments, R ² and R ³ can be connected to form a substituted or unsubstituted cyclopropyl ring. In some embodiments, R 2 and R ³ can be connected to form an unsubstituted cyclopropyl ring. In some embodiments, R ² and R 3 can be connected to form a substituted or unsubstituted cyclobutyl ring. In some embodiments, R ² and R ³ can be connected to form an unsubstituted cyclobutyl ring. In some embodiments, R ² and R ³ can be connected to form a substituted cyclobutyl ring. In some embodiments, R ² and R ³ can be connected to form a substituted cyclobutyl ring. In some embodiments, R ² and R ³ can be connected to form a substituted cyclobutyl ring of the following formula: In some embodiments, R ² and R ³ can be connected to form a substituted or unsubstituted cyclopentyl ring. In some embodiments, ^R and ^R can be connected to form an unsubstituted cyclopentyl ring. In some embodiments, ^R and ^R can be connected to form a substituted or unsubstituted monocyclic or bicyclic 3 to 8 membered heterocycle. In some embodiments, ^R and ^R can be connected to form a substituted or unsubstituted 3 to 9 membered heterocycle (e.g., a substituted or unsubstituted 5 to 9 membered monocyclic heterocycle containing 0, 1 or 2 double bonds in the heterocyclic ring system, wherein 1, 2 or 3 atoms in the heterocyclic ring system are independently nitrogen, oxygen or sulfur). In some embodiments, ^R and ^R can be connected to form a substituted or unsubstituted tetrahydropyran ring.

In some embodiments, R ⁴ and/or R ⁵ can be hydrogen. In some embodiments, R ⁴ and/or R ⁵ can be substituted or unsubstituted C _1-6 alkyl (e.g., substituted or unsubstituted methyl, ethyl, propyl, or butyl). In some embodiments, R ⁴ and/or R ⁵ can be substituted methyl. In some embodiments, R ⁴ and/or R ⁵ can be unsubstituted methyl. In some embodiments, R ⁴ and/or R ⁵ can be substituted ethyl. In some embodiments, R ⁴ and/or R ⁵ can be unsubstituted ethyl. In some embodiments, R ⁴ and/or R ⁵ can be propyl. In some embodiments, R ⁴ and/or R ⁵ can be unsubstituted isopropyl. In some embodiments, R ⁴ and/or R ⁵ can be isobutyl. In some embodiments, R ⁴ and/or R ⁵ can be the following formula: wherein R can be substituted or unsubstituted C _1-6 alkyl. In some embodiments, R can be substituted or unsubstituted methyl, ethyl, propyl, or butyl. In some embodiments, R can be -CF ₃ . In some embodiments, R ⁴ and/or R ⁵ can be substituted or unsubstituted C ₂ -C ₆ alkenyl. In some embodiments, R ⁴ and/or R ⁵ can be substituted or unsubstituted C ₅ -C ₈ cycloalkenyl. In some embodiments, R ⁴ and/or R ⁵ can be substituted or unsubstituted C ₂ -C ₁₀ alkynyl (e.g., substituted or unsubstituted propynyl or butynyl). In some embodiments, R ⁴ and/or R ⁵ can be substituted or unsubstituted aryl (e.g., phenyl or benzyl). In some embodiments, R ⁴ can be of the following formula: In some embodiments, R ⁴ and/or R ⁵ can be of the following formula: In some embodiments, R ⁴ and/or R ⁵ can be substituted or unsubstituted C ₁ -C ₆ alkoxy (e.g., substituted or unsubstituted methoxy or ethoxy). In some embodiments, R ⁴ and/or R ⁵ can be substituted or unsubstituted C ₃ -C ₈ cycloalkyl (e.g., substituted or unsubstituted cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In some embodiments, R ⁴ and/or R ⁵ can be of the following formula: In some embodiments, R ⁴ and/or R ⁵ can be substituted or unsubstituted 3 to 12-membered heterocyclyl (e.g., substituted or unsubstituted 3 to 12-membered monocyclic or bicyclic heterocyclyl, which contains 0, 1, or 2 double bonds in the heterocyclic ring system, wherein 1, 2, or 3 atoms in the heterocyclic ring system are independently nitrogen, oxygen, or sulfur). In some embodiments, R ⁴ and/or R ⁵ can be of the following formula: In some embodiments, R ⁴ and/or R ⁵ can be substituted or unsubstituted 5 to 6-membered monocyclic heteroaryl, wherein 1, 2, 3, or 4 atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur. In some embodiments, ^R4 and/or ^R5 can be substituted or unsubstituted 8- to 10-membered bicyclic heteroaryl, wherein one, two, three, or four atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur. In some embodiments, ^R4 and/or ^R5 can be substituted or unsubstituted aryl ( _C1 - _C6 alkyl). In some embodiments, ^R4 and/or ^R5 can be arylsulfonyl.

In some embodiments, ^R4 and ^R5 are independently one of the following formulae:

In some embodiments, ^R4 and ^R5 are independently one of the following formulae:

In some embodiments, ^R and ^R can be connected together with the N to which they are connected to form optionally substituted monocyclic or bicyclic heterocyclic radicals. In some embodiments, ^R and ^R can be connected together with the N to which they are connected to form optionally substituted 5 to 7 yuan of heterocyclic radicals. In some embodiments, ^R and ^R can be connected together with the N to which they are connected to form optionally substituted 6 yuan of heterocyclic radicals. In some embodiments, ^R and ^R can be connected together with the N to which they are connected to form optionally substituted 6 yuan of heterocyclic radicals containing 1 or 2 heteroatoms independently selected from N, S and O. In certain embodiments, ^R and ^R can be connected together with the N to which they are connected to form optionally substituted piperidines. In certain embodiments, ^R and ^R can be connected together with the N to which they are connected to form optionally substituted morpholine. For example, in certain embodiments, ^R and ^R can be connected together with the N to which they are connected to form one of the following:

In some embodiments, ^R6 in formula (I) can be a substituted or unsubstituted _C1-6 alkyl group (e.g., substituted or unsubstituted methyl, ethyl, propyl, or butyl). In some embodiments, ^R6 can be an isopropyl group. In some embodiments, ^R6 can be a substituted methyl group. In some embodiments, ^R6 can be In some embodiments, ^R6 can be a substituted or unsubstituted _C3 - _C8 cycloalkyl group (e.g., substituted or unsubstituted cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In some embodiments, ^R6 can be In some embodiments, ^R6 can be a substituted or unsubstituted benzoyl group. In some embodiments, ^R6 can be a substituted or unsubstituted _C2 - _C6 alkenyl group. In some embodiments, ^R6 can be a substituted or unsubstituted _C2 -C6 alkynyl group. In some embodiments, ^R6 can be a substituted or unsubstituted _C5 - _{C8 cycloalkenyl} group. In some embodiments, ^R6 can be a substituted or unsubstituted aryl group (e.g., phenyl or benzyl). In some embodiments, R ⁶ can be substituted or unsubstituted benzyl. In some embodiments, R ⁶ can be the following formula: wherein R ^6A can be hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, halogen, -CN, -OR ^6a or substituted or unsubstituted sulfonyl, wherein R ^6a can be hydrogen or substituted or unsubstituted C ₁ -C ₆ alkyl; and k can be 0, 1 or 2. In some embodiments, R ^6a can be substituted or unsubstituted C ₁ -C ₆ alkyl (e.g., methyl, ethyl, propyl or butyl). In some embodiments, R ^6A can be halogen (e.g., F, Cl, Br or I). In some embodiments, R ^6a can be -CN. In some embodiments, k can be 0. In some embodiments, k can be 1. In some embodiments, k can be 2.

In some embodiments, ^R6 can be of the following formula: In some embodiments, ^R6 can be a substituted or unsubstituted 4 to 7 membered (e.g., 4, 5, 6, or 7) monocyclic heterocyclyl group containing 0, 1, or 2 double bonds in the heterocyclic ring system, wherein 1, 2, or 3 atoms in the heterocyclic ring system are independently nitrogen, oxygen, or sulfur. In some embodiments, ^R6 can be In some embodiments, ^R6 can be a substituted or unsubstituted 7 to 10 membered bicyclic heterocyclyl group containing 0, 1, or 2 double bonds in the heterocyclic ring system, wherein 1, 2, or 3 atoms in the heterocyclic ring system are independently nitrogen, oxygen, or sulfur. In some embodiments, ^R6 can be a substituted or unsubstituted 5 to 6 membered monocyclic heteroaryl group, wherein 1, 2, 3, or 4 atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur. In some embodiments, R ⁶ can be of the following formula: In some embodiments, R ⁶ can be of the following formula: In some embodiments, R ⁶ can be a substituted or unsubstituted 8- to 10-membered bicyclic heteroaryl group, wherein 1, 2, 3, or 4 atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur. In some embodiments, R ⁶ can be of the following formula: In some embodiments, R ⁶ can be a substituted or unsubstituted C ₁ -C ₆ alkoxy group (e.g., a substituted or unsubstituted methoxy or ethoxy group). In some embodiments, R ⁶ can be a substituted or unsubstituted aryloxy group. In some embodiments, R ⁶ can be -C(=O)R ⁷ , wherein R ⁷ is as defined herein. In some embodiments, R 6 can be

In some embodiments, the compound of formula (I) can be a compound of one of the following formulae: formula (II), formula (III), formula (IV), formula (V), or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, tautomer, stereoisomer, isotopically labeled derivative or prodrug thereof.

In some embodiments, the compound of formula (II) can be a compound of formula 1-30, 59, 62-67, 83, 85, 90, a compound described herein, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, tautomer, stereoisomer, isotopically labeled derivative or prodrug thereof.

In some embodiments, the compound of formula (III) can be a compound of formula 31-35, a compound described herein, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, tautomer, stereoisomer, isotopically labeled derivative or prodrug thereof.

In some embodiments, the compound of formula (IV) can be a compound of formula 36-46, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, tautomer, stereoisomer, isotopically labeled derivative or prodrug thereof.

In some embodiments, the compound of formula (V) can be compounds 47-58, 60-61, 68-82, 84, 86-89, 91-125 and 126-128 described herein, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, tautomer, stereoisomer, isotopically labeled derivative or prodrug thereof.

Exemplary IDO-inhibiting compounds as described herein and their characterizations are provided in Table 1 below:

Table 1. Characterization of compounds of formula (I)

The compounds described herein can be prepared from readily available raw materials using methods known in the art. It should be understood that, where typical or preferred process conditions (i.e., reaction temperature, time, molar ratio of reactants, solvent and pressure, etc.) are given, other process conditions may also be used unless otherwise stated. Optimal reaction conditions may vary with the specific reactants or solvents used, but these conditions can be determined by those skilled in the art through conventional optimization procedures. The chemicals used in the above-mentioned synthetic routes may include, for example, solvents, reagents, catalysts, protecting groups, and deprotecting group reagents. The above-mentioned methods may also further include steps of adding or removing suitable protecting groups before or after the steps specifically described herein, so as to ultimately synthesize the compound. In addition, the various synthetic steps may be performed in different orders or sequences to obtain the desired compound. Synthetic chemistry transformations and protecting group methods (protection and deprotection) used to synthesize suitable compounds are known in the art and include, for example, those described in the following books: R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T. W. Greene and P. G. Wuts, Protective Groups in Organic Synthesis, ^3rd Ed., John Wiley and Sons (1999); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995) and subsequent editions.

The compounds of formula (I) provided herein can be prepared from readily available raw materials using the following general methods and procedures. An exemplary schematic diagram for synthesizing the compounds of the present invention described herein is provided below. Where typical or preferred process conditions (i.e., reaction temperature, time, molar ratio of reactants, solvent, pressure, etc.) are given, other process conditions may also be used unless otherwise indicated. Optimal reaction conditions may vary with the specific reactants or solvents used, but these conditions may be determined by conventional optimization procedures by those skilled in the art.

The compounds of the present invention can be prepared according to General Route A. Compounds A1, where G = halogen, are commercially available or can be synthesized by standard transformations known to those of ordinary skill in the art of organic/medicinal chemistry. Compound ^A2 can be prepared by replacing the halogen in A1 with an amine ^HNR4R5 in a solvent such as THF, DMF, NMP, etc., using base or palladium catalysis. Reduction of the nitro group can be carried out in a solvent such as methanol or ethyl acetate under reducing conditions, such as, but not limited to, palladium on carbon, under a hydrogen atmosphere, to provide intermediate A3. Treatment of aniline A3 with isocyanate A4 in a solvent such as THF at a temperature between ambient temperature and the boiling point of the solvent provides intermediate A5. Nitrile A5 can be hydrolyzed under acidic or basic conditions to prepare Ia. Alternatively, nitrile A5 can be converted to tetrazole Ib by heating it with an azide such as _NaN3 , _TMSN3 , or tributyltin azide in a solvent such as toluene at or near boiling point.

Route A. Preparation of compounds of formula (I)

Route B. Preparation of acid derivative I-a

Route B shows an alternative way to convert intermediate A3 into acid derivative I-a. Nitrile A3 can be hydrolyzed to prepare B1 under acidic or basic conditions. Aniline B1 is treated with isocyanate A4 in a solvent (e.g., THF) at a temperature between ambient temperature and the boiling point of the solvent to give I-a.

Route C. Preparation of compounds of formula (I)

Compounds of the present invention can also be prepared according to General Route C. Compounds C1, wherein G = halogen, are commercially available or can be synthesized by standard transformations known to those of ordinary skill in the art of organic/pharmaceutical chemistry. Compound C2 can be prepared by replacing the halogen in C1 with an amine ^HNR4R5 in a solvent such as THF, DMF, NMP, etc., by base or ^palladium catalysis. Reduction of the nitro group can be carried out in a solvent such as methanol or ethyl acetate under reducing conditions such as, but not limited to, palladium on carbon under a hydrogen atmosphere to provide intermediate C3. Treatment of aniline C3 with isocyanate A4 in a solvent such as THF at a temperature between ambient temperature and the boiling point of the solvent provides intermediate C4. Saponification of C4 to Ia can generally be accomplished using an alkali metal hydroxide in an aqueous or mixed aqueous/organic solvent.

Route D. Preparation of compounds of formula (I)

The compounds of the present invention can also be prepared according to General Route D. Compounds D1 wherein G = halogen are commercially available or can be synthesized by standard transformations known to those skilled in the art of organic/pharmaceutical chemistry. Compound D2 can be prepared by replacing the halogen in D1 with an amine ^HNR4R5 by base or palladium catalysis in a solvent such as THF, DMF, NMP, ^etc. Palladium-catalyzed cross-coupling can produce thioether D3. Reduction of the nitro group can be carried out in a solvent such as methanol or ethyl acetate under reducing conditions such as, but not limited to, palladium on carbon under a hydrogen atmosphere to give intermediate D4. Aniline D4 is treated with isocyanate A4 in a solvent such as THF at a temperature between ambient temperature and the boiling point of the solvent to give intermediate D5. Deprotection of thioether D5 can provide thiol D6, which can be converted to ester derivative D7 under displacement conditions. Saponification of D7 to IV-a can generally be accomplished by using an alkali metal hydroxide in an aqueous or mixed aqueous/organic solvent.

Route E. Preparation of compounds of formula (I)

Referring to Scheme E, compounds E1, wherein V is CN or an ester, can be prepared using the above transformations. Aniline E1 can be converted to an isothiocyanate by treatment with a reagent such as thionyl chloride. Treatment of the isothiocyanate with o-phenylenediamine (o-dimines) followed by heating the reaction in the presence of a base can form benzimidazole E3. Saponification of E3 to V-a can typically be accomplished using an alkali metal hydroxide in an aqueous or mixed aqueous/organic solvent.

Route F. Preparation of compounds of formula (I)

In Scheme F, compound F1 can be prepared from amine E1 by halogen displacement of XR ¹⁸ (via base or palladium catalysis). Saponification of F1 to Vb can generally be accomplished using alkali metal hydroxides in aqueous or mixed aqueous/organic solvents.

Route G. Preparation of compounds of formula (I)

In Scheme G, compound F1 can be prepared from amine E1 by amide formation (e.g., treatment with an acid chloride in the presence of a base). Saponification of G1 to V-c can generally be accomplished using an alkali metal hydroxide in an aqueous or mixed aqueous/organic solvent.

Pharmaceutical compositions and kits

The present disclosure provides a pharmaceutical composition comprising a compound as described herein or a pharmaceutically acceptable salt thereof, and an optional pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition as described herein comprises a compound as described herein or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. The pharmaceutical composition as described herein can be used to treat and/or prevent proliferative diseases (such as cancer) and infectious diseases (such as viral or bacterial infectious diseases). In some instances, the pharmaceutical composition as described herein may further comprise a second therapeutic agent, such as those described herein, such as an anticancer agent or an antiviral agent.

In certain embodiments, the cells contacted with an effective amount of a compound or pharmaceutical composition as described herein are in vitro. In certain embodiments, the cells contacted are ex vivo. In certain embodiments, the cells as described herein are in vivo. In certain embodiments, the cells as described herein are malignant cells (e.g., malignant blood cells).

In certain embodiments, the compound as described herein is provided in an effective amount in a pharmaceutical composition. In certain embodiments, an effective amount is a therapeutically effective amount (for example, effectively treating the amount of a proliferative disease of a subject in need thereof). In certain embodiments, the proliferative disease is cancer. In certain embodiments, the proliferative disease is cancer, such as non-small cell lung cancer, small cell lung cancer, breast cancer, renal cell carcinoma, bladder cancer, head and neck cancer, ovarian cancer, brain cancer, gastrointestinal cancer, liver cancer, pancreatic cancer, melanoma, leukemia, lymphoma, etc. In certain embodiments, an effective amount is a preventive effective amount (for example, effectively preventing the proliferative disease of a subject in need thereof and/or alleviating the amount of a proliferative disease of a subject in need thereof).

The pharmaceutical compositions described herein can be prepared by any method known in the pharmaceutical industry. Generally, such preparation methods include combining the compound described herein (i.e., the "active ingredient") with a carrier or excipient and/or one or more other auxiliary ingredients, and then, if necessary and/or desired, shaping and/or packaging the product into desired single or multiple dose units.

Pharmaceutical compositions can be prepared, packaged, and/or sold in bulk as a single unit dose, and/or as a plurality of single unit doses. A "unit dose" is a discrete amount of a pharmaceutical composition containing a predetermined amount of an active ingredient. The amount of the active ingredient is generally equal to the dose of the active ingredient to be administered to a subject and/or a convenient fraction of such a dose, such as one-half or one-third of such a dose.

The relative amounts of the active ingredient, pharmaceutically acceptable excipients, and/or any additional ingredients in the pharmaceutical compositions described herein will vary depending on the identity, size, and/or condition of the subject being treated, and further vary depending on the route by which the composition is to be administered. The composition may contain 0.1% to 100% (w/w) active ingredient.

Pharmaceutically acceptable excipients used in the preparation of the provided pharmaceutical compositions include inert diluents, dispersants and/or granulating agents, surfactants and/or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants and/or oils. Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweeteners, flavorings, and fragrances may also be present in the compositions.

Liquid dosage forms for oral and parenteral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, the liquid dosage form may also contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof. In addition to inert diluents, oral compositions may also contain adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavorings, and aromatics. In certain embodiments for parenteral administration, the conjugates described herein are mixed with solubilizers (e.g., alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and mixtures thereof).

Injectable preparations, such as sterile injectable aqueous or oily suspensions, can be prepared using suitable dispersants or wetting agents and suspending agents according to known techniques. Sterile injectable preparations can be sterile injectable solutions, suspensions or emulsions in a non-toxic parenterally acceptable diluent or solvent, such as a solution in 1,3-butanediol. Among acceptable carriers and solvents, water, Ringer's solution, U.S.P. and isotonic sodium chloride solution can be used. In addition, sterile fixed oils are generally used as solvents or suspending media. For this purpose, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids (such as oleic acid) are used in the preparation of injections.

For example, the injectable formulation can be sterilized by filtration through a bacteria-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active ingredient is mixed with at least one inert pharmaceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate and/or (a) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia, (c) humectants, such as glycerol, (d) disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, (e) solution retarding agents, such as paraffin, (f) absorption promoters, such as quaternary ammonium compounds, (g) wetting agents, such as, for example, cetyl alcohol and glyceryl monostearate, (h) adsorbents, such as kaolin and bentonite, and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may comprise buffering agents.

The solid composition of similar type can be used as the filler in soft and hard filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol. The solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coating and shell (for example other coatings well-known to enteric coatings and pharmacological fields). They can optionally include an opacifying agent and can have a composition that makes it, optionally in a delayed manner, only in or preferentially releases active ingredient in a certain part of intestinal tract. The example of operable encapsulated composition includes polymeric substances and wax. The solid composition of similar type can be used as the filler in soft and hard filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol.

Active ingredient can be made into microencapsulated form with one or more excipients as described above. The solid dosage forms of tablets, lozenges, capsules, pills and granules can be prepared with coatings and shells (e.g., enteric coatings, release-controlled coatings and other coatings known in the field of pharmaceutical formulations). In such solid dosage forms, the active ingredient can be mixed with at least one inert diluent (e.g., sucrose, lactose or starch). Normally, such dosage forms can include other substances other than inert diluents, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage form can include a buffer. They can optionally include an opacifying agent and can have a composition that allows them to, optionally in a delayed manner, only release the active ingredient in or preferentially in a certain part of the intestinal tract. The example of an encapsulating agent that can be used includes polymeric substances and waxes.

Suitable devices for delivering intradermal pharmaceutical compositions as herein described include short needle devices. Intradermal compositions can be used by limiting the effective penetration length of the needle entering the skin. Alternatively or additionally, conventional syringes can be used in the classical Mantoux method of intradermal administration. It is suitable to deliver liquid formulations to the jet injection device of the dermis via a liquid jet injector and/or via a needle that pierces the stratum corneum and produces a jet that arrives the dermis. It is suitable to utilize compressed gas to accelerate the compound in powder form through the skin outer layer to arrive the ballistic powder/particle delivery device of the dermis.

Although the descriptions of pharmaceutical compositions provided herein are primarily directed to pharmaceutical compositions suitable for administration to humans, such compositions are generally suitable for administration to various animals. Modifications to pharmaceutical compositions suitable for administration to humans are well understood, and a veterinary pharmacologist of ordinary skill can design and/or perform such modifications using ordinary experimentation.

The compounds provided herein are generally formulated in dosage unit form for ease of administration and uniformity of dosage. However, it should be understood that the total daily dosage of the compositions described herein will be determined by a physician within the scope of sound medical judgment. The specific therapeutically effective dosage level for any particular subject or organism will depend on a variety of factors, including the disease being treated and the severity of the disease; the activity of the specific active ingredient used; the specific composition employed; the age, weight, general health, sex, and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific active ingredient used; the duration of treatment; drugs used in combination or concomitantly with the specific active ingredient used; and similar factors well known in the medical field.

The present disclosure also includes kits (e.g., pharmaceutical packaging). The kits provided may include pharmaceutical compositions or compounds described herein and containers (e.g., vials, ampoules, bottles, syringes, and/or subpackages, or other suitable containers). In some embodiments, the kits provided may optionally further include a second container comprising a pharmaceutical excipient for diluting or suspending the pharmaceutical compositions or compounds described herein. In some embodiments, the pharmaceutical compositions or compounds described herein provided in the first container and the second container are combined to form a unit dosage form.

In certain embodiments, the kits described herein include a first container containing a compound or pharmaceutical composition as described herein. In certain embodiments, the kits described herein can be used to treat a proliferative disease (e.g., non-small cell lung cancer, small cell lung cancer, breast cancer, renal cell carcinoma, bladder cancer, head and neck cancer, ovarian cancer, brain cancer, gastrointestinal cancer, liver cancer, pancreatic cancer, melanoma, leukemia, lymphoma, etc.) in a subject in need thereof, and/or to prevent a proliferative disease in a subject in need thereof.

In certain embodiments, the kits described herein further include instructions for use of the compound or pharmaceutical composition contained in the kit. Kits described herein may also include information required by regulatory agencies such as the U.S. Food and Drug Administration (FDA). In certain embodiments, the information contained in the kit is prescription information. In certain embodiments, the kit and instructions are provided for treating a proliferative disease in a subject in need thereof, and/or preventing a proliferative disease in a subject in need thereof. Kits described herein may include one or more additional agents as described herein as additional compositions.

Treatment

As shown in the following examples, the exemplary IDO inhibitory compounds described herein successfully demonstrated in vitro efficacy and in vivo efficacy. The compounds described herein can be used to treat and/or prevent proliferative diseases (e.g., cancer) by inhibiting IDO and inhibiting tryptophan catabolism, resulting in reduced kynurenine levels. In addition, compared to other IDO inhibitors known in the art (including INCB-24360 and other compounds disclosed in WO2014150677 and WO2014150646), these compounds show lower human liver clearance. Therefore, the present disclosure provides methods for treating diseases associated with IDO with one or more IDO inhibitory compounds described herein. Diseases associated with IDO include, but are not limited to, cancer, infectious diseases, and Alzheimer's disease. In certain embodiments, the infectious disease is a viral infection.

Accordingly, the present disclosure provides methods of treating a proliferative disease in a subject in need thereof, comprising administering to the subject an effective amount (eg, a therapeutically effective amount) of a compound described herein or a pharmaceutical composition thereof.

Another aspect of the present disclosure relates to methods of preventing a proliferative disease in a subject in need thereof, comprising administering to the subject an effective amount (eg, a prophylactically effective amount) of a compound described herein or a pharmaceutical composition thereof.

Compounds and pharmaceutical compositions as described herein can be used to treat and/or prevent proliferative diseases. In certain embodiments, the proliferative disease is cancer (e.g., lung cancer (non-small cell lung cancer, small cell lung cancer), breast cancer, prostate cancer, ovarian cancer, endometrial cancer, cervical cancer, bladder cancer, head and neck cancer, renal cell carcinoma, esophageal cancer, pancreatic cancer, brain cancer, gastrointestinal cancer, liver cancer, leukemia, lymphoma, melanoma, multiple myeloma, Ewing's sarcoma or osteosarcoma). In certain embodiments, the proliferative disease is an inflammatory disease. In certain embodiments, the proliferative disease is an immune-related disease.

In certain embodiments, the methods described herein further include administering another agent to the subject. In certain embodiments, the methods described herein further include contacting the biological sample with another agent. In certain embodiments, the methods described herein further include contacting the tissue with another agent. In certain embodiments, the methods described herein further include treating the subject in need of treatment with a second anticancer therapy (e.g., chemotherapy, immunotherapy (e.g., anti-PD-1 or anti-PD-L1 antibodies), cell therapy (e.g., CAR-T cell therapy), surgery and/or transplantation (e.g., bone marrow transplantation)). In some instances, the second anticancer therapy includes the use of one or more anticancer agents, such as anticancer agents known in the art, including anticancer drugs in clinical use or clinical trials.

The compounds and compositions provided herein can be administered by any route, including enteral (e.g., oral), parenteral and/or intravenous. Specific considerations include oral administration, intravenous administration (e.g., systemic intravenous injection), regional administration by blood and/or lymphatic supply, and/or administration directly to the affected area. Typically, the most appropriate route of administration will depend on various factors, including the properties of the drug (e.g., its stability in a gastrointestinal environment), and/or the condition of the subject (e.g., whether the subject can tolerate oral administration).

The exact amount of compound required to reach an effective amount will vary from subject to subject, depending on, for example, the species, age, and general condition of the subject, the severity of the side effects or disorder, the characteristics of the specific compound, the mode of administration, etc. An effective amount may be included in a single dose (e.g., a single oral dose) or multiple doses (e.g., multiple oral doses). In certain embodiments, when multiple doses are administered to a subject or applied to a biological sample, tissue, or cell, any two doses of the multiple doses include different or substantially the same amount of a compound as described herein. In certain embodiments, when multiple doses are administered to a subject or applied to a biological sample, tissue, or cell, the frequency of administering multiple doses to the subject or applying multiple doses to the tissue or cell is two doses a day, one dose a day, one dose every other day, one dose every three days, one dose a week, one dose every two weeks, one dose every three weeks, or one dose every four weeks. In certain embodiments, the frequency of administering multiple doses to the subject or applying multiple doses to the tissue or cell is one dose a day. In certain embodiments, the frequency of administering multiple doses to the subject or applying multiple doses to the tissue or cell is two doses a day. In certain embodiments, the frequency of applying multiple doses to a subject or applying multiple doses to a tissue or cell is three doses per day. In certain embodiments, when multiple doses are applied to a subject or applied to a biological sample, tissue or cell, the duration between the first dose of multiple doses and the last dose is half a day, one day, two days, four days, one week, two weeks, three weeks, one month, two months, three months, four months, six months, nine months, one year, two years, three years, four years, five years, seven years, ten years, fifteen years, twenty years or the life span of the subject, biological sample, tissue or cell. In certain embodiments, the duration between the first dose of multiple doses and the last dose is three months, six months or one year. In certain embodiments, the duration between the first dose of multiple doses and the last dose is the life span of the subject, biological sample, tissue or cell. In certain embodiments, the doses described herein (e.g., a single dose, or any dose in a multiple dose) independently include between 0.1 μg and 1 μg, between 0.001 mg and 0.01 mg, between 0.01 mg and 0.1 mg, between 0.1 mg and 1 mg, between 1 mg and 3 mg, between 3 mg and 10 mg, between 10 mg and 30 mg, between 30 mg and 100 mg, between 100 mg and 300 mg, between 300 mg and 1,000 mg, or between 1 g and 10 mg (including end values) of a compound described herein. In certain embodiments, the doses described herein independently include between 3 mg and 10 mg (including end values) of a compound described herein. In certain embodiments, the doses described herein independently include between 10 mg and 30 mg (including end values) of a compound described herein. In certain embodiments, the doses described herein independently include between 30 mg and 100 mg (including end values) of a compound described herein. In certain embodiments, the dosages described herein independently include a compound described herein between 100 mg and 300 mg (including end values). In certain embodiments, the dosages described herein independently include a compound described herein between 300 mg and 1000 mg (including end values).

The dosage ranges described herein provide guidance for administering the provided pharmaceutical compositions to adults. For example, the amount administered to a child or adolescent can be determined by a physician or person skilled in the art and can be lower than or equal to the amount administered to an adult.

Compound as herein described or composition can be used for treating and/or preventing one or more other medicaments (for example, treatment and/or prevention active agent) combination administration for proliferative disease.Described compound or composition can be used for treating and/or preventing one or more other medicaments (for example, treatment and/or prevention active agent) combination administration for proliferative disease.Described compound or composition can be used for treating and/or preventing one or more other medicaments (for example, treatment and/or prevention active agent) combination administration for proliferative disease for the subject of these compounds or compositions, described other medicament improves the activity (for example, treatment has the proliferative disease of the subject in need and/or prevention has the activity (for example, effect and/or efficacy) of the proliferative disease of the subject in need), improves bioavailability, improves safety, reduces drug resistance, reduces and/or changes metabolism, suppresses excretion and/or changes the distribution in subject, biological sample, tissue or cell.It will also be understood that the therapy adopted can realize the desired effect to the same illness, and/or it can realize different effects.In certain embodiments, the pharmaceutical composition as herein described comprising compound as herein described and other medicament is presented at the synergistic effect that is not present in the pharmaceutical composition comprising one of described compound and other medicament and not comprising both.

The compound or composition can be used simultaneously with one or more other medicaments, and is used before or after it, and the one or more other medicaments can be used for example for the conjoint therapy of treating and/or preventing proliferative diseases. Medicament includes therapeutic active agent. Medicament also includes preventive active agent. Medicament includes small organic molecules, such as pharmaceutical compounds (compounds approved for human or veterinary use by the U.S. Food and Drug Administration as specified in the U.S. Federal Regulations (CFR)), peptides, proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucins, lipoproteins, synthetic polypeptides or proteins, small molecules connected to proteins, glycoproteins, steroids, nucleic acids, DNA, RNA, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins and cells. In certain embodiments, the other medicament is an agent that can be used to treat proliferative diseases. In certain embodiments, the other medicament is an agent that can be used to prevent proliferative diseases. In certain embodiments, the other medicament is an agent approved for treating and/or preventing proliferative diseases by management agencies (e.g., U.S. FDA). In some embodiments, the combination of the present invention can be used in a dosage and/or timetable determined for the agent. The other agent can also be used together with each other and/or with the compounds or compositions described herein in a single dose or separately in different doses. The specific combination used in the scheme will take into account the compatibility of the compounds described herein with the other agent and/or the desired treatment and/or preventive effect to be achieved. In general, it is expected that the use level of the other agent in the combination will not exceed the level at which they are used alone. In some embodiments, the level used in the combination will be lower than the level used alone.

In certain embodiments, the other medicament is an antiproliferative agent (such as an anticancer agent). In certain embodiments, the other medicament is an anticancer agent, an anti-angiogenic agent, an anti-inflammatory agent, an immunosuppressant, an antibacterial agent, an antiviral agent, a cardiovascular agent, a cholesterol-lowering agent, an antidiabetic agent, an antiallergic agent, an analgesic or a combination thereof. In certain embodiments, the compound as described herein or pharmaceutical composition can be administered in combination with anticancer therapy, and the anticancer therapy includes but is not limited to transplantation (such as bone marrow transplantation, stem cell transplantation), surgery, radiotherapy, cell therapy (such as CAR-T cell therapy), immunotherapy (such as anti-PD-1 or anti-PD-L1 antibodies or cancer vaccines) and chemotherapy. Treatment with IDO inhibition compounds can be performed before, simultaneously or afterwards in another therapy.

When any of the IDO inhibitory compounds described herein are used to treat a viral infection, they can be used in conjunction with a second antiviral agent, which can be different from any of the IDO inhibitory compounds described herein. In some examples, the antiviral agent is an antiviral vaccine. The second antiviral agent can be administered before, simultaneously with, or after administration of the IDO inhibitory compound.

As known in the art, other combination therapies involving IDO inhibitors are also within the scope of the present disclosure. See, for example, WO2015006520, the relevant disclosure of which is incorporated by reference for the purposes or subject matter cited herein.

Without further elaboration, it is believed that one skilled in the art can utilize the present invention to its fullest extent based on the above description. Therefore, the following detailed description is to be construed as merely illustrative and not limiting in any way the remainder of the present disclosure. All publications cited herein are incorporated herein by reference for the purposes or subject matter cited herein.

Example

In order that the present disclosure may be more fully understood, the following examples are set forth.The synthetic and biological examples described in this application are provided to illustrate the compounds, pharmaceutical compositions and methods provided herein and should not be construed as limiting the scope thereof in any way.

Example 1

Step 1. Synthesis of 1-1

A solution of 2-(4-fluorophenyl)acetonitrile (5 g, 37.00 mmol) in sulfuric acid (50 mL) was placed in a 100 mL three-necked round-bottom flask. Potassium nitrate (5.6 g) was then added portionwise at 0°C over 10 minutes. The resulting solution was stirred at room temperature overnight. The reaction mixture was poured into 150 mL of water/ice. The resulting solution was extracted with 3×100 mL of ethyl acetate. The combined organic layers were washed with 3×100 mL of brine, dried over anhydrous sodium sulfate and concentrated in vacuo to give 2-(4-fluoro-3-nitrophenyl)acetamide (6 g, 82% yield).

Step 2. Synthesis of 1-2

A solution of 2-(4-fluoro-3-nitrophenyl)acetamide (6 g, 30.28 mmol) in DMSO (60 mL), bis(2-methylpropyl)amine (5.86 g, 45.34 mmol), and DIEA (7.81 g, 60.66 mmol) were placed in a 250 mL three-necked round-bottom flask. The resulting solution was heated to 100°C and stirred at the same temperature overnight. The reaction mixture was cooled to room temperature and then quenched by the addition of 50 mL of water. The resulting solution was extracted with 3 × 100 mL of ethyl acetate. The combined organic layers were washed with 3 × 100 mL of brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:10 to 1:3) to give 2-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]acetamide (7 g, 75% yield).

Step 3. Synthesis of 1-3

A solution of 2-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]acetamide (7 g, 22.77 mmol) in dioxane (70 mL), TFAA (7 mL) and triethylamine (3 mL) were placed in a 250 mL three-necked round-bottom flask. The resulting solution was heated to 100 ° C and stirred at the same temperature overnight. The reaction mixture was cooled to room temperature. 50 mL of water was then added to quench the reaction. The resulting solution was extracted with 3×100 mL of ethyl acetate. The combined organic layers were washed with 3×100 mL of brine, dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:20 to 1:3) to give 2-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]acetonitrile (6.1 g, 93% yield).

Step 4. Synthesis of 1-4

A solution of 2-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]acetonitrile (3 g, 10.37 mmol) in ethanol/ _H₂O (30/10 mL), Fe (3.49 g), and _NH₄Cl (380 mg, 7.10 mmol) were placed in a 100 mL round-bottom flask. The resulting solution was stirred at 80°C for 2 hours. The reaction mixture was cooled to room temperature. The resulting mixture was concentrated in vacuo. The resulting solution was extracted with 3 x 50 mL of ethyl acetate. The combined organic layers were washed with 3 x 50 mL of brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:10-1:1) to provide 2-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]acetonitrile (1.1 g, 41% yield).

Step 5. Synthesis of 1-5

A solution of 2-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]acetonitrile (1.1 g, 4.24 mmol) in tetrahydrofuran (50 mL), 2,4-difluoro-1-isocyanatophenyl ester (790 mg, 5.09 mmol), and triethylamine (860 mg, 8.50 mmol) were placed in a 100-mL three-necked round-bottom flask. The resulting solution was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:10 to 1:3) to provide 3-[2-[bis(2-methylpropyl)amino]-5-(cyanomethyl)phenyl]-1-(2,4-difluorophenyl)urea (700 mg, 40% yield).

Step 6. Synthesis of 1

A solution of 1-[2-[bis(2-methylpropyl)amino]-5-(cyanomethyl)phenyl]-3-(2,4-difluorophenyl)urea (300 mg, 0.72 mmol) in ethanol/ _H₂O (5/1 mL) and sodium hydroxide (15% in water) (1 mL) were placed in an 8 mL sealed tube. The resulting solution was stirred at 60°C overnight. The reaction mixture was cooled to room temperature and concentrated in vacuo. The pH of the solution was adjusted to 6 with hydrogen chloride ( ₁ N). The resulting mixture was concentrated in vacuo. The crude product was purified by preparative HPLC using the following conditions: column, Waters X-bridge RP18, 19 x 150 mm, 5 μm; mobile phase, ACN/water (0.05% _NH₃H₂O ) 20% to 38% over 5.6 minutes, flow rate: 20 mL/min; detector, 254 nm. Yielded 72.2 mg (23% yield) of 2-[4-[bis(2-methylpropyl)amino]-3-[[(2,4-difluorophenyl)carbamoyl]amino]phenyl]acetic acid as an off-white solid. LCMS(ES,m/z):434[M+H] ⁺ .HNMR(300MHz,DMSO-d ₆ ,ppm): δ9.27(s,1H),8.06(s,1H),7.96-7.88(m,1H),7.78(d,J＝1.8Hz,1H),7.33-7.25(m,1H),7.12(d,J＝8.1Hz,1H),7.0 7-7.00(m,1H),6.86(dd,J＝8.4,2.1Hz,1H),3.38(s,2H),2.66(d,J＝6.9Hz,4H),1.69-1.61(m,2H),0.84(d,J＝6.6Hz,12H).

Example 2

Step 1. Synthesis of 2-1

To a solution of 2-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]acetonitrile (2 g, 6.91 mmol) in tetrahydrofuran (20 mL) was added sodium hydride (250 mg, 6.25 mmol) in portions at 0°C. After stirring at room temperature for 1 hour, the reaction was cooled to 0°C and iodomethane (1.18 g, 8.31 mmol) was added dropwise. The reaction mixture was then stirred at room temperature for another 2 hours and then quenched by the addition of saturated ammonium chloride solution (20 mL). The mixture was extracted with ethyl acetate (50 mL×3) and washed with brine (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by Flash-Prep-HPLC [column: C18; mobile phase A: water (0.05% ammonium bicarbonate), mobile phase B: acetonitrile; gradient: 35% acetonitrile-78% acetonitrile in 30 minutes] to give the desired product (660 mg, 31% yield).

Step 2. Synthesis of 2-2

To a solution of 2-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]propionitrile (600 mg, 1.98 mmol) in ethanol (20 mL) was added palladium on carbon (300 mg). The reaction was stirred at room temperature under a balloon _{of H2} for 2 hours. The solid was filtered off and the filtrate was concentrated in vacuo to give the desired product (530 mg, 72% yield).

Step 3. Synthesis of 2-3

To a solution of 2-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]propionitrile (546 mg, 2.00 mmol) in tetrahydrofuran (20 mL) was added triethylamine (610 mg, 6.03 mmol) and 2,4-difluoro-1-isocyanatophenyl ester (456 mg, 2.94 mmol). After stirring at room temperature for 3 hours, the reaction was concentrated in vacuo and the residue was purified by Flash-Prep-HPLC [column: C18; mobile phase A: water (0.05% ammonium bicarbonate), mobile phase B: acetonitrile; gradient: 45% acetonitrile to 85% acetonitrile over 30 minutes] to give the desired product (450 mg, 40% yield).

Step 4. Synthesis of 2

To a solution of 3-[2-[bis(2-methylpropyl)amino]-5-(1-cyanoethyl)phenyl]-1-(2,4-difluorophenyl)urea (60 mg, 0.14 mmol) in ethanol (4 mL) and water (1 mL) was added sodium hydroxide (400 mg, 10.00 mmol). After stirring at 60 ° C for 16 hours, the reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was dissolved in water (10 mL) and the pH was adjusted to 4 using hydrogen chloride (4N). The mixture was then extracted with ethyl acetate (20 mL × 3). The organic phase was washed with brine (20 mL × 3), dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by preparative HPLC [column: Waters X-bridge C18, 19×150 mm; mobile phase A: water (0.05% trifluoroacetic acid), mobile phase B: acetonitrile; gradient: 25% acetonitrile to 60% acetonitrile; 8 minutes; detector: 254 nm] to give the desired product (15.8 mg, 25% yield). LCMS(ES,m/z):448.3[M+H] ⁺ ; ¹ HNMR: (300MHz, DMSO-d ₆ ,ppm): δ9.29(s,1H),8.06(s,1H),7.93-7.91(m,1H),7.84(d,J＝1.8Hz,1H),7.35-7.25(m,1H),7.17-7.11(m,1H),7.06-6.97(m,1 H),6.91-6.88(m,1H),3.58-3.51(m,1H),2.66(d,J＝6.9Hz,4H),1.69-1.61(m,2H),1.32(d,J＝7.2Hz,3H),0.84(d,J＝6.6Hz,12H).

Example 3

Step 1. Synthesis of 3-1

At 0 ℃, sodium hydride (3.91g, 97.75mmol) was added to a solution of 2-(4-fluorophenyl)acetonitrile (11g, 81.40mmol) in tetrahydrofuran (250mL). The mixture was then heated to room temperature and stirred for 30 minutes. The reaction mixture was cooled to 0 ℃ again and bromoethane (9.76g, 89.57mmol) was added. The reaction was then stirred at room temperature overnight. Water (100mL) was added and the mixture was extracted with ethyl acetate (100mL×2). The organic layer was dried over anhydrous sodium sulfate and concentrated in vacuo. Ethyl acetate/petroleum ether (1/10) was used as eluent to pass through a silica gel column purification residue to obtain 2-(4-fluorophenyl)butyronitrile (9g, 83% yield).

Step 2: Synthesis of 3-2

At 0 ° C, potassium nitrate (8.28 g, 82.82 mmol) was added to a solution of 2- (4-fluorophenyl) butyronitrile (9 g, 55.21 mmol) in sulfuric acid (200 mL). The reaction was then stirred at 25 ° C for 1 hour. Water/ice (600 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (100 mL × 3). The organic layer was washed with saturated sodium bicarbonate aqueous solution (30 mL × 2), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/4) as eluent to obtain 2- (4-fluoro-3-nitrophenyl) butyronitrile (5 g, 44% yield).

Step 3. Synthesis of 3-3

To a solution of 2-(4-fluoro-3-nitrophenyl)butyronitrile (5 g, 24 mmol) and diisopropylethylamine (6.2 g, 48 mmol) in dimethyl sulfoxide (100 mL) was added bis(2-methylpropyl)amine (3.7 g, 28.8 mmol). The mixture was then heated to 100° C. and stirred for 1 hour. After cooling to room temperature, ethyl acetate (200 mL) was added and the mixture was washed with water (100 mL×2). The organic layer was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/10) to give 2-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]butyronitrile (4 g, 53% yield).

Step 4. Synthesis of 3-4

Under hydrogen atmosphere (1 atmosphere), at 25 ℃, a mixture of 2-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]butyronitrile (4g, 12.60mmol) and palladium on carbon (0.4g) in methanol (20mL) was stirred overnight. The reactant was filtered through diatomaceous earth and the filtrate was concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/10) to give 2-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]butyronitrile (3g, 83% yield).

Step 5. Synthesis of 3-5

At 100 ° C, a mixture of 2-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]butyronitrile (1.16 g, 4.04 mmol) and potassium hydroxide (1.13 g, 20.14 mmol) in ethanol (10 mL) and water (2 mL) was stirred in a sealed tube for 3 days. The pH value of the solution was adjusted to 7 with hydrogen chloride (2N). Water (50 mL) was added and the mixture was extracted with ethyl acetate (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give 4-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]tetrahydropyran-4-carboxylic acid (1.16 g, 94% yield).

Step 6. Synthesis of 3-6

A mixture of 2-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]butanoic acid (1.16 g, 3.79 mmol), 2,4-difluoro-1-isocyanatophenyl ester (707 mg, 4.56 mmol), and triethylamine (768 mg, 7.59 mmol) in tetrahydrofuran (10 mL) was stirred overnight at 25° C. The mixture was concentrated and the residue was purified by preparative HPLC to give the desired product in racemic form. (Column: X Bridge RP, 5 μm, 19×150 mm; Mobile phase A: 10 mmol/L NH ₄ HCO ₃ in water; Mobile phase B: ACN; Flow rate: 30 mL/min; Gradient: 45% B to 85% B over 8 minutes; Detector: 254 nm).

Step 7. Chiral separation

The racemic product from step 6 was separated by chiral preparative HPLC under the following conditions to give Examples 3A and 3B (column: IA, 20 mmd x 250 mmd; mobile phase: 3% ethanol in hexane; flow rate: 15 mL/min; detector: 254 nm).

Example 3A: Retention time: 30.07 minutes. LRMS:(ES,m/z):462.4[M+H] ⁺ . ¹ HNMR:(300MHz,DMSO-d ₆ ): δ12.22(s,1H),9.30(s,1H),8.07(s,1H),7.97-7.89(m,1H),7.84(s,1H),7.33-7.25(m,1H),7.14(d,J＝8.4,1H),7.06-6.99( m,1H),6.88(d,J＝6.9Hz,1H),3.30(t,J＝7.8,1H),2.65(d,J＝6.3,4H),1.97–1.85(m,1H),1.70–1.55(m,3H),0.83–0.78(m,15H).

Example 3B: Retention time: 38.62 minutes. LRMS(ES,m/z):462.4[M+H] ⁺ . ¹ HNMR:(300MHz,DMSO-d ₆ ): δ12.20(s,1H),9.30(s,1H),8.07(s,1H),7.97-7.89(m,1H),7.83(s,1H),7.35–7.27(m,1H),7.15(d,J＝8.4,1H),7.07–7.0 1(m,1H),6.90–6.87(m,1H),3.33–3.27(m,1H),2.66(d,J＝6.6,4H),1.97–1.87(m,1H),1.69–1.57(m,3H),0.85–0.80(m,15H).

Example 4

Step 1. Synthesis of 4-1

At 0 ° C, sodium hydride (250 mg, 10.37 mmol) was added in tetrahydrofuran (7 mL) solution of 2-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]acetonitrile (0.7 g, 3.46 mmol) in batches. The mixture was then stirred at 0 ° C for 1 hour and iodomethane (0.858 g, 6.05 mmol) was added. After stirring at room temperature for another 16 hours, the reaction was quenched by adding saturated ammonium chloride solution. The mixture was extracted with ethyl acetate (50 mL × 2) and washed with brine (50 mL × 2). The combined organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column using ethyl acetate/petroleum ether (1/15) as eluent to obtain the desired product (300 mg, 39% yield).

Compound 4 was synthesized according to the similar procedure as in Example 2.

Example 4: LRMS (ES, m/z): 462.40 [M+H] ⁺ ; ¹ H-NMR: (300MHz, DMSO-d ₆ ,ppm): δ12.05(brs,1H),9.27(s,1H),8.06(s,1H),7.96-7.88(m,2H),7.33.7.26(m,1H),7.15-7.12(m ,1H),7.07-6.94(m,2H),2.65(d,J＝6.9Hz,4H),1.69-1.60(m,2H),1.42(s,6H),0.84(d,J＝6.6Hz,12H)

Example 5

Step 1. Synthesis of 5

To a solution of azidotrimethylsilane (185 mg, 1.61 mmol) in toluene (15 mL) was added a solution of chlorodiethylalumane (194 mg, 1.61 mmol) in toluene (1.8 mL). After stirring at room temperature for 6 hours, a solution of 3-[2-[bis(2-methylpropyl)amino]-5-(1-cyanoethyl)phenyl]-1-(2,4-difluorophenyl)urea (230 mg, 0.54 mmol) in toluene (20 mL) was added. The resulting mixture was stirred at 110 ° C for another 16 hours. The reactants were cooled to room temperature and concentrated in vacuo. The residue was dissolved in ethyl acetate (50 mL), and the mixture was washed with brine (30 mL×3). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by preparative HPLC under the following conditions [column: Waters X-bridge C18, 19×150 nm; mobile phase A: water (0.05% ammonia), mobile phase B: acetonitrile; gradient: 25% acetonitrile to 52% acetonitrile; 6.8 minutes, 25 mL/min; detector, 254 nm]. The collected fractions were combined and concentrated in vacuo to give the desired racemic product (60 mg, 24% yield) as a white solid.

Step 2. Chiral separation

The racemate (60 mg) was separated by chiral-preparative HPLC under the following conditions [column: AD-H; mobile phase A: 5% hexane, mobile phase B: ethanol; gradient: 5% ethanol; 25 mL/min; detector, 254 nm]. The collected fractions were combined and concentrated in vacuo to give the desired products 5A (15.3 mg, 26% yield, RT = 4.33 min) and 5B (13.3 mg, 22% yield, RT = 5.45 min) as white solids.

Compound 5A: LCMS (ES, m/z): 472.5[M+H] ⁺ . ¹ HNMR (300MHz, DMSO-d ₆ ): δ9.30(s,1H),8.06(s,1H),7.95-7.86(m,1H),7.80(d,J＝1.8Hz,1H),7.33-7.28(m,1H),7.16-7.14(m,1H),7.07-7.01 (m,1H),6.85(dd,J＝8.4,1.8Hz,1H),4.45-4.42(m,1H),2.65(d,J＝6.6Hz,4H),1.70-1.59(m,5H),0.83(d,J＝6.6Hz,12H).

Compound 5B: LCMS (ES, m/z): 472.5[M+H] ⁺ . ¹ HNMR (300MHz, DMSO-d ₆ ): δ9.29(s,1H),8.05(s,1H),7.95-7.87(m,1H),7.80(d,J＝2.1Hz,1H),7.33-7.26(m,1H),7.15-7.12(m,1H),7.07-7.01 (m,1H),6.85(dd,J＝8.4,1.8Hz,1H),4.44-4.38(m,1H),2.64(d,J＝6.6Hz,4H),1.67-1.56(m,5H),0.83(d,J＝6.6Hz,12H).

Example 6

Step 1. Synthesis of 6-1

To a solution of 2-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]acetonitrile (2.0 g, 6.91 mmol) in acetonitrile (30 mL) was added selective fluorine reagent (Selectfluor) (4.9 g, 13.84 mmol) in portions. The resulting mixture was stirred at room temperature for 6 hours. The reactants were then concentrated in vacuo. The residue was purified by silica gel column using ethyl acetate/petroleum ether (1/10) as eluent to give the desired product (0.55 g, 26%).

Step 2. Synthesis of 6-2

To a solution of 2-[4-[bis(2-methylpropyl)amino]-3-fluoro-5-nitrophenyl]acetonitrile (550 mg, 1.79 mmol) in acetic acid (5 mL) was added iron (1.0 g, 17.86 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was filtered through diatomaceous earth and the filtrate was concentrated in vacuo. The residue was dissolved in ethyl acetate (50 mL) and washed with saturated sodium bicarbonate aqueous solution (20 mL) and water (20 mL). The organic phase was concentrated in vacuo and the residue was purified by silica gel column purification using ethyl acetate/petroleum ether (1/10) as eluent to obtain the desired product (0.4 g, 81%).

Step 3. Synthesis of 6-3

To a solution of 2-[3-amino-4-[bis(2-methylpropyl)amino]-5-fluorophenyl]acetonitrile (190 mg, 0.68 mmol) in tetrahydrofuran (3 mL) was added 2,4-difluoro-1-isocyanatophenyl ester (160 mg, 1.03 mmol) and triethylamine (0.21 g, 2.04 mmol). The mixture was then stirred at room temperature for 3 hours. The reactant was diluted with ethyl acetate (20 mL) and then washed with water (10 mL × 2). The organic phase was concentrated in vacuo. The residue was purified by silica gel column using ethyl acetate/petroleum ether (1/2) as eluent to obtain the desired product (0.14 g, 47%).

Step 4. Synthesis of 6

A solution of 3-[2-[bis(2-methylpropyl)amino]-5-(cyanomethyl)-3-fluorophenyl]-1-(2,4-difluorophenyl)urea (140 mg, 0.32 mmol), trimethylsilyl azide (300 mg, 2.60 mmol), and tetrabutylammonium fluoride (500 mg, 1.91 mmol) in toluene (3 mL) was stirred at 90° C. for 1 hour. The mixture was cooled to room temperature and diluted with ethyl acetate (20 mL). The mixture was washed with water (10 mL x 2). The organic phase was concentrated in vacuo. The residue (420 mg) was purified by preparative HPLC under the following conditions: [column, X Bridge Shield RP18 OBD column, 5 um, 19×150 mm; mobile phase, water (0.05% TFA) and ACN/MeOH (15%-60.0% in 8 minutes); detector, UV 254 nm] to give the desired product (36.2 mg, 24%) as an off-white solid. LCMS(ES,m/z):476.4[M+H] ⁺ ; ¹ HNMR: (300MHz, DMSO-d ₆ ,ppm): δ9.43(s,1H),8.34(s,1H),7.93-7.84(m,1H),7.79(s,1H),7.73-7.68(m,1H),7.34-7.26(m,1H),7.07-7 .05(m,1H),6.67(dd,J＝11.4Hz,1.5Hz,1H),4.11(s,2H),3.51-3.42(m,2H),2.79(m,4H),0.83(d,J＝4.5Hz,12H).

Example 7

Step 1. Synthesis of 7-2

To a solution of 2-(4-fluorophenyl)acetonitrile (8 g, 59.20 mmol) in toluene (160 mL) was added 1,2-dibromoethane (11.28 g, 60.04 mmol), potassium hydroxide (27 g, 481.20 mmol, 8.00 equivalents), tetrabutylammonium bromide (200 mg, 0.62 mmol) and water (8 mL) at 0 ° C. The reaction mixture was heated to 100 ° C and stirred for 1.5 hours. The reaction was cooled, quenched with water (100 mL), and extracted with ethyl acetate (100 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/10) as eluent to give 1-(4-fluorophenyl)cyclopropane-1-carbonitrile (5 g, 52%).

Step 2. Synthesis of 7-3

At 0 ℃, potassium nitrate (4.7g, 46.49mmol) was added to a solution of 1-(4-fluorophenyl)cyclopropane-1-carbonitrile (5g, 31.02mmol) in sulfuric acid (100mL). The mixture was then heated to room temperature and stirred for 40 minutes. The reaction was quenched with water/ice (300mL) and the mixture was extracted with ethyl acetate (100mL×3). The organic layer was washed with saturated sodium bicarbonate aqueous solution (30mL×3), dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/4) as eluent to obtain 1-(4-fluoro-3-nitrophenyl)cyclopropane-1-carbonitrile (1.6g, 25%).

Step 3. Synthesis of 7-4

To a solution of 1-(4-fluoro-3-nitrophenyl)cyclopropane-1-carbonitrile (1.6 g, 7.76 mmol) and N,N-diisopropylethylamine (2 g, 15.48 mmol) in dimethyl sulfoxide (50 mL) was added bis(2-methylpropyl)amine (1.2 g, 9.28 mmol). The mixture was stirred at 100 ° C for 2.5 hours. The reaction was cooled, quenched with water (150 mL), and extracted with ethyl acetate (100 mL×3). The organic layer was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/8) to give 1-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]cyclopropane-1-carbonitrile (1.94 g, 79%).

Step 4. Synthesis of 7-5

Under hydrogen atmosphere (1 atmosphere), at room temperature, a mixture of 1-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]cyclopropane-1-carbonitrile (1.9 g, 6.02 mmol) and palladium on carbon (0.19 g) in methanol (20 mL) was stirred for 2 hours. The mixture was filtered through celite and the filtrate was concentrated in vacuo. The residue was purified by silica gel column purification with ethyl acetate/petroleum ether (1/8) to give 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]cyclopropane-1-carbonitrile (1.0 g, 58%).

Step 5. Synthesis of 7-6

A mixture of 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]cyclopropane-1-carbonitrile (500 mg, 1.75 mmol) and potassium hydroxide (5 mL, 3 M in _H₂O ) in ethanol (10 mL) was stirred in a sealed tube overnight at 100°C. After cooling to room temperature, the pH of the solution was adjusted to 7 with aqueous hydrogen chloride (2N), and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]cyclopropane-1-carboxylic acid (530 mg, 99%).

Step 6. Synthesis of 7

To a solution of 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]cyclopropane-1-carboxylic acid (530 mg, 1.74 mmol) and 2,4-difluoro-1-isocyanatophenyl ester (324 mg, 2.09 mmol) in tetrahydrofuran (353 mg, 4.89 mmol) was added triethylamine (5 mL) and the reaction mixture was stirred at 25° C. for 3 hours. The resulting mixture was concentrated in vacuo and then purified by preparative HPLC (column: XBridge Shield RP18 OBD column, 5 μm, 19×150 mm; mobile phase A: water containing 0.05% NH ₄ HCO ₃ , mobile phase B: ACN; flow rate: 25 mL/min; gradient: 25% B-85% B in 8 minutes; detector: UV 254 nm) to give 1-[4-[bis(2-methylpropyl)amino]-3-[[(2,4-difluorophenyl)carbamoyl]amino]phenyl]cyclopropane-1-carboxylic acid (146.8 mg, 18% yield). LRMS: (ES, m/z): [M+H] ⁺ =460.3. ¹ H NMR (300 MHz, DMSO-d ₆ ): δ12.25(s,br,1H),9.29(s,1H),8.04(s,1H),7.96-7.85(m,1H),7.84(s,1H),7.34-7.26(m,1H),7.10(d,J＝8.1Hz,1H),7.09-6.93( m,1H),6.91(d,J＝2.1Hz,1H),2.65(d,J＝6.9Hz,4H),1.70-1.59(m,2H),1.42-1.39(m,2H),1.09-1.05(m,2H),0.86(d,J＝6.6Hz,12H).

Example 8

Step 1. Synthesis of 8-2

At 0 ° C, sodium hydride (3.7 g, 154 mmol) was added to a solution of 2-(4-fluorophenyl)acetonitrile (5 g, 37 mmol) in N, N-dimethylformamide (20 mL). Then heated to room temperature and stirred for 30 minutes. The reaction mixture was cooled to 0 ° C again and 1-bromo-2-(2-bromoethoxy)ethane (8.59 g, 37 mmol) was added. Then the reaction was stirred at room temperature overnight. Water (100 mL) was added and the mixture was extracted with ethyl acetate (50 mL × 3). The organic layer was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/9) as eluent to obtain 4-(4-fluorophenyl)tetrahydropyran-4-carbonitrile (3.69 g, 49%).

Step 2. Synthesis of 8-3

At 0 ° C, potassium nitrate (2.73 g, 27.00 mmol) was added to a solution of 4-(4-fluorophenyl)tetrahydropyran-4-carbonitrile (3.69 g, 17.98 mmol) in sulfuric acid (30 mL). The reaction mixture was then stirred at 25 ° C for 1 hour. Water/ice (250 mL) was added to quench the reaction and the mixture was extracted with ethyl acetate (50 mL×3). The organic layer was washed with saturated sodium bicarbonate aqueous solution (20 mL×2), dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/9) as eluent to give 4-(4-fluoro-3-nitrophenyl)tetrahydropyran-4-carbonitrile (2.38 g, 53%).

Step 3. Synthesis of 8-4

To a solution of 4-(4-fluoro-3-nitrophenyl)tetrahydropyran-4-carbonitrile (2.30 g, 9.19 mmol) and diisopropylethylamine (2.37 g, 18.37 mmol) in dimethyl sulfoxide (10 mL) was added bis(2-methylpropyl)amine (1.42 g, 11.02 mmol). The reaction mixture was then heated to 100° C. and stirred for 4.5 hours. Water (100 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (100 mL×3). The organic layer was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/5) to give 4-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]tetrahydropyran-4-carbonitrile (2.7 g, 82%).

Step 4. Synthesis of 8-5

Under a hydrogen atmosphere (1 atm), a mixture of 4-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]tetrahydropyran-4-carbonitrile (2.7 g, 7.51 mmol) and palladium on carbon (0.3 g) in methanol (50 mL) was stirred for 4.5 hours at 25° C. The reaction mixture was then filtered through celite and the filtrate was concentrated in vacuo. The residue was purified on a silica gel column with ethyl acetate/petroleum ether (1/4) to give 4-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]tetrahydropyran-4-carbonitrile (1.62 g, 65%).

Step 5. Synthesis of 8-6

A mixture of 4-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]tetrahydropyran-4-carbonitrile (500 mg, 1.52 mmol) and potassium hydroxide (3 M in H ₂ O, 5 mL) in ethanol (10 mL) was stirred in a sealed tube at 100° C. for 2 days. Water (50 mL) was added and the mixture was extracted with ethyl acetate (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give 4-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]tetrahydropyran-4-carboxylic acid (520 mg, 98%).

Step 6. Synthesis of 8

A mixture of 4-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]tetrahydropyran-4-carboxylic acid (520 mg, 1.49 mmol), 2,4-difluoro-1-isocyanatophenyl ester (279 mg, 1.80 mmol) and triethylamine (303 mg, 2.99 mmol) in tetrahydrofuran (5 mL) was stirred for 3 hours at 25° C. Water (20 mL) was added and the mixture was extracted with ethyl acetate (50 mL×3). The organic layer was concentrated and the residue was purified by preparative HPLC (column: X Bridge Shield RP18 OBD column, 5 um, 19×150 mm; mobile phase A: water containing 0.05% NH ₄ HCO ₃ , mobile phase B: ACN; flow rate: 25 mL/min; gradient: 25% B to 85% B in 8 minutes; detector: UV; 254 nm) to give 4-[4-[bis(2-methylpropyl)amino]-3-[[(2,4-difluorophenyl)carbamoyl]amino]phenyl]tetrahydropyran-4-carboxylic acid (88.56 mg, 12%) as a white solid. LRMS(ES,m/z):[M+H] ⁺ =504.5. ¹ HNMR(300MHz,DMSO-d ₆ )δ9.30(s,1H),8.04(s,1H),7.97-7.89(m,2H),7.34-7.26(m,1H),7.15(d,J＝8.4Hz,1H),7.07-6.98(m,2H),3.80(d,J＝ 11.7,2H),3.54-3.41(m,2H),2.66(d,J＝6.9Hz,4H),2.34(d,J＝13.2Hz,2H),1.75-1.60,(m,4H),0.84(d,J＝6.6Hz,12H).

Example 9

Step 1. Synthesis of 9-1

A solution of 2-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]acetonitrile (5 g, 17.28 mmol) in DMSO (100 mL) was placed in a 250 mL three-necked round-bottom flask, and sodium hydride (2.77 g, 69.25 mmol) was added portionwise at 10 ° C. The resulting mixture was stirred at room temperature for 1 hour. 1,4-dibromobutane (7.47 g, 34.60 mmol) was added dropwise with stirring at 10 ° C. over 5 minutes. The resulting solution was stirred at room temperature overnight. The reaction was then quenched by adding 50 mL of water and extracted with 3×100 mL of ethyl acetate. The combined organic layers were washed with 3×100 mL of brine. The mixture was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:10 to 1:3) to give 1-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]cyclopentane-1-carbonitrile (2.5 g, 42% yield).

Step 2. Synthesis of 9-2

Methanol (10 mL), 1-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]cyclopentane-1-carbonitrile (1 g, 2.91 mmol) and sulfuric acid (5 mL) were placed in a 50 mL round-bottom flask. The resulting solution was heated at reflux for 48 hours. The reaction was cooled to room temperature. The pH of the solution was adjusted to 8 with sodium carbonate (10% aqueous solution). The resulting mixture was extracted with 3×50 mL of ethyl acetate. The combined organic phases were washed with 3×50 mL of brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:20 to 1:5) to give 1-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]cyclopentane-1-carboxylic acid methyl ester (450 mg, 41% yield).

Step 3. Synthesis of 9-3

To 1-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]cyclopentane-1-carboxylic acid methyl esters (450mg, 1.20mmol) in methanol (5mL) solution, palladium carbon (500mg) is added. The flask is evacuated and flushed with nitrogen three times, then flushed with hydrogen. The mixture is stirred for 3 hours under a hydrogen atmosphere (balloon) at room temperature. The solid is filtered off. The resulting mixture is concentrated in vacuo. With ethyl acetate/petroleum ether (1:10-1:1), the residue is applied to a silica gel column to obtain 1-(3-amino-4-(diisobutylamino)phenyl)cyclopentanecarboxylic acid methyl esters (350mg, 85% yield).

Step 4. Synthesis of 9-4

Tetrahydrofuran (5 mL), methyl 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]cyclopentane-1-carboxylate (350 mg, 1.01 mmol), triethylamine (202 mg, 2.00 mmol) and phenyl 2,4-difluoro-1-isocyanate (188 mg, 1.21 mmol) were placed in a 25 mL round-bottom flask. The resulting solution was stirred at room temperature for 4 hours. The reaction was concentrated under vacuum. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:10-1:3) to give methyl 1-[4-[bis(2-methylpropyl)amino]-3-[[(2,4-(difluorophenyl)carbamoyl]amino]phenyl]cyclopentane-1-carboxylate (250 mg, 49% yield).

Step 5. Synthesis of 9

A solution of methyl 1-[4-[bis(2-methylpropyl)amino]-3-[[(2,4-difluorophenyl)carbamoyl]amino]phenyl]cyclopentane-1-carboxylate (250 mg, 0.50 mmol) in methanol (5 mL) and sodium hydroxide (1 mL, 15% aqueous solution) were placed in a 25 mL round-bottom flask. The resulting solution was stirred at room temperature for 4 hours. The resulting mixture was concentrated under vacuum. The pH of the solution was adjusted to 6 with hydrogen chloride (1N). The resulting solution was extracted with 3×10 mL of ethyl acetate. The combined organic layers were washed with 3×10 mL of brine and concentrated under vacuum. The crude product was purified by Flash-Prep-HPLC under the following conditions: column: SunFire C18 5um 19×150mm; mobile phase; CH ₃ CN/water (0.05% NH ₃ H ₂ O; gradient: 51%-65% in 7 minutes; flow rate: 20 mL/min; detector, UV 254 nm) to give 79.5 mg (33% yield) of 1-[4-[bis(2-methylpropyl)amino]-3-[[(2,4-difluorophenyl)carbamoyl]amino]phenyl]cyclopentane-1-carboxylic acid as an off-white solid. LCMS:(ES,m/z):488.3[M+H] ⁺ .HNMR:(300MHz,DMSO-d ₆ ,ppm): δ12.10(s,1H),9.25(s,1H),8.03(s,1H),7.95-7.87(m,2H),7.31(t,J＝2.7Hz,1H),7.12(d,J＝8.4Hz,1H),7.07-7.0 0(m,1H),6.94(dd,J＝8.4,2.4Hz,1H),2.65(d,J＝6.9Hz,4H),2.49-2.45(m,2H),1.76-1.58(m,8H),0.84(d,J＝6.6Hz,12H).

Example 10

Step 1. Synthesis of 10-1

A solution of 1-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]cyclopentane-1-carbonitrile (1.5 g, 4.37 mmol) in ethanol (30 mL), Fe (1.47 g, 26.25 mmol), and _NH₄Cl (162 mg, 3.03 mmol) were placed in a 100 mL round-bottom flask. The resulting solution was stirred at 80°C for 3 hours. The reaction mixture was cooled to room temperature. The solid was filtered off, and the filtrate was concentrated in vacuo. The residue was dissolved in 150 mL of ethyl acetate, washed with water (50 mL) and brine (3 x 50 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:10-1:1) to provide 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]cyclopentane-1-carbonitrile (600 mg, 44% yield).

Step 2. Synthesis of 10-2

Tetrahydrofuran (10 mL), triethylamine (387 mg, 3.82 mmol), and 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]cyclopentane-1-carbonitrile (600 mg, 1.91 mmol) were placed in a 50 mL three-necked round-bottom flask. 2,4-difluoro-1-phenylisocyanate (356 mg, 2.30 mmol) was then added dropwise with stirring at 0°C. The resulting solution was stirred at room temperature for 3 hours. The reaction was concentrated in vacuo. The residue was applied to a silica gel column using ethyl acetate/petroleum ether (1:20 to 1:3) to provide 3-[2-[bis(2-methylpropyl)amino]-5-(1-cyanocyclopentyl)phenyl]-1-(2,4-difluorophenyl)urea (500 mg, 56% yield).

Step 3. Synthesis of 10

Sodium azide (62 mg, 0.95 mmol) and toluene (3 mL) are placed in a 50 mL 2-neck round-bottom flask purged and maintained with a nitrogen inert atmosphere. Subsequently, diethylaluminum chloride (0.9 M) (1.07 mL) is added dropwise under stirring at 0 ° C. The resulting solution is stirred at room temperature for 7 hours. A toluene (1 mL) solution of 1- [2- [bis (2- methylpropyl) amino] -5- (1- cyanocyclopentyl) phenyl] -3- (2,4- difluorophenyl) urea (300 mg, 0.64 mmol) is added thereto. The resulting solution is heated and stirred at 120 ° C overnight. The reactant is cooled to room temperature and 10 mL of water is added to quench it. The resulting solution is extracted with 3 × 50 mL of ethyl acetate. The combined organic layer is washed with 3 × 50 mL of brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The crude product was purified by preparative HPLC under the following conditions: column, Waters Sunfire C18 19×150 mm 5 μm; mobile phase, CH ₃ CN/water (0.05% TFA), 65%-74% (7 minutes); detector, 254 nm. This gave 60 mg (15% yield) of 3-[2-[bis(2-methylpropyl)amino]-5-[1-(1H-1,2,3,4-tetrazol-5-yl)cyclopentyl]phenyl]-1-(2,4-difluorophenyl)urea as a trifluoroacetic acid salt. LCMS.(ES,m/z):512[M+H-CF ₃ COOH] ⁺ .HNMR(300MHz,DMSO-d ₆ ,ppm)δ9.29(s,1H),8.02(s,1H),7.94-7.86(m,2H),7.30(t,J＝6.0Hz,1H),7.27(d,J＝6.3Hz,1H),7.11-7.02(m,1H),6.85(dd,J＝8.4 ,2.1Hz,1H),2.73-2.63(m,6H),2.18-2.13(m,2H),1.76-1.66(m,2H),1.66-1.60(m,2H),1.58-1.49(m,2H),0.82(d,J＝6.6Hz,12H).

Example 11

Step 1. Synthesis of 11-1

At 0 ° C, sodium hydride (17.8 g, 440 mmol) was added portionwise to a solution of 2- (4-fluorophenyl) acetonitrile (20.0 g, 148 mmol) in tetrahydrofuran (200 mL). The mixture was then stirred at 0 ° C for 30 minutes. A solution of 1,4-dibromobutane (38.4 g, 178 mmol) in tetrahydrofuran (100 mL) was added dropwise, and the reaction mixture was stirred at room temperature for another 4 hours. The reactant was then cooled to 0 ° C and water (10 mL) was added to quench the reaction. The resulting mixture was diluted with ethyl acetate (1 L) and washed with water (600 mL) and brine (600 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate / petroleum ether (1/8) as eluent to obtain the desired product (22.1 g, 71% yield).

Step 2. Synthesis of 11-2

At 0 ° C, potassium nitrate (17.7 g, 175 mmol) was added in batches to a solution of 1-(4-fluorophenyl)cyclopentanecarbonitrile (22.1 g, 117 mmol) in concentrated sulfuric acid (200 mL). The resulting mixture was then stirred at 0 ° C for 10 minutes. The reactant was carefully poured into ice water (1 L). The mixture was extracted with ethyl acetate (600 mL × 2), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/6) as eluent to obtain the desired product (15.2 g, 55% yield).

Step 3. Synthesis of 11-3

At 0 ℃, 2-methylpropanoyl chloride (5g, 46.93mmol) was added dropwise to a solution of cyclohexylamine (5g, 50.42mmol) and triethylamine (8mL) in tetrahydrofuran (45mL). The mixture was then stirred at room temperature for 3 days. The reaction was quenched by adding saturated ammonium chloride (150mL) and extracted with dichloromethane (150mL). The combined organic phases were washed with salt water (200mL×2), dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain the desired product (7.5g, 88% yield).

Step 4. Synthesis of 11-4

At 0 ℃, lithium aluminum tetrahydride (6.5 g, 171.28 mmol) was added to a solution of N-cyclohexyl-2-methylpropionamide (7 g, 41.36 mmol) in tetrahydrofuran (140 mL) several times. The mixture was then stirred at 70 ℃ overnight. The mixture was cooled to 0 ℃ and quenched by adding saturated ammonium chloride (250 mL). The solid was filtered off and the filter cake was washed with ethyl acetate (200 mL × 2). The combined organic phase was separated and washed with brine (300 mL × 2), dried over anhydrous sodium sulfate, and concentrated in vacuo to give the desired product (4 g, 62% yield).

Step 5. Synthesis of 11-5

To a solution of 1-(4-fluoro-3-nitrophenyl)cyclopentane-1-carbonitrile (1g, 4.27mmol) in dimethyl sulfoxide (20mL) was added N-(2-methylpropyl)cyclohexylamine (1g, 6.44mmol) and N,N-diisopropylethylamine (1.65g, 12.77mmol) in sequence. The mixture was then stirred at 100°C for 16 hours. The reactant was cooled to room temperature and diluted with water (100mL). The mixture was extracted with ethyl acetate (100mL×3) and washed with brine (100mL×5). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/99) to obtain the desired product (1g, 63% yield).

Step 6. Synthesis of 11-6

To 1-[4-[cyclohexyl(2-methylpropyl)amino]-3-nitrophenyl]cyclopentane-1-carbonitrile (1g, 2.71mmol) in acetic acid (10mL) solution was added iron (1.5g, 26.86mmol). The mixture was then stirred at room temperature for 1 hour. Ethyl acetate (150mL) was added and the solid was filtered out. The filtrate was diluted with water (100mL) and the pH value was adjusted to 9 by adding saturated sodium bicarbonate aqueous solution. The organic phase was separated and washed with brine (100mL×2), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/25-1/20) to obtain the desired product (597mg, 65% yield).

Step 7. Synthesis of 11-7

To a solution of 1-[3-amino-4-[cyclohexyl(2-methylpropyl)amino]phenyl]cyclopentane-1-carbonitrile (200 mg, 0.589 mmol) in ethanol (10 mL) and water (2 mL) was added potassium hydroxide (3 g, 53.47 mmol). The mixture was then stirred at 100°C for 3 days. The mixture was cooled to room temperature and concentrated in vacuo. The residue was dissolved in water (50 mL) and the pH was adjusted to 6 with hydrogen chloride (1 N). The mixture was extracted with dichloromethane (50 mL x 2) and washed with brine (30 mL x 2). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by Flash-Prep-HPLC under the following conditions: [column, C18 silica gel; mobile phase (A: MeCN, B: H ₂ O (0.1% TFA), MeCN = 50%; detector, UV 254 nm) to give the desired product (85 mg, 40% yield).

Step 8. Synthesis of 11

To a solution of 1-[3-amino-4-[cyclohexyl(2-methylpropyl)amino]phenyl]cyclopentane-1-carboxylic acid (85 mg, 0.24 mmol) in tetrahydrofuran (50 mL) were added 2,4-difluoro-1-isocyanatophenyl ester (51 mg, 0.33 mmol) and triethylamine (111 mg, 1.10 mmol) in sequence. The reaction was then stirred at room temperature for 2 hours. The mixture was concentrated in vacuo. The residue was redissolved in water (50 mL) and the pH was adjusted to 6 with HCl (1N). The mixture was extracted with dichloromethane (50 mL × 2) and washed with brine (30 mL × 2). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by Flash-Prep-HPLC under the following conditions: [column, C18 silica gel; mobile phase (A: MeCN, B: H ₂ O), MeCN = 96%; detector, UV 254 nm] to give the desired product (26.6 mg, 22% yield) as a white solid. LCMS (ES, m/z): 514.50 [M+1] ⁺ ; ¹ H NMR (300 MHz, DMSO-D ₆ ,ppm): δ12.23(s,1H),9.37(t,J＝4.6Hz,1H),8.14(s,1H),7.99(d,J＝2.2Hz,1H),7.90(td,J＝9.2,6.2Hz,1H),7.32(td,J＝8.8,4.5Hz,1H),7 .22–6.99(m,2H),6.99–6.85(m,1H),2.76(d,J＝7.3Hz,2H),1.90-1.80(m,2H),1.77–1.47(m,9H),1.40–1.00(m,9H),0.82(d,J＝6.5Hz,6H).

Example 12

Step 1. Synthesis of 12-1

At 100 ℃, 1-(4-fluoro-3-nitrophenyl) cyclopentane-1-carbonitrile (1.5g, 6.40mmol), 4,4-difluorocyclohexane-1-amine (950mg, 7.03mmol) and N, N-diisopropylethylamine (1.65g) in dimethyl sulfoxide (20mL) solution was stirred overnight. The mixture was cooled to room temperature, diluted with ethyl acetate (80mL), and washed with water (60mL) and brine (60mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. Ethyl acetate/petroleum ether (1/10) was used as eluent to pass through a silica gel column purification residue to obtain the desired product (2.1g, 94% yield).

Step 2. Synthesis of 12-2

At 0 ° C, sodium hydride (720 mg, 18.00 mmol) was added in batches to a solution of 1-[4-[(4,4-difluorocyclohexyl)amino]-3-nitrophenyl]cyclopentane-1-carbonitrile (2.1 g, 6.01 mmol) in tetrahydrofuran (25 mL). The mixture was then stirred at 0 ° C for 30 minutes, followed by a solution of 3-bromo-2-methylprop-1-ene (1.22 g, 9.04 mmol) in tetrahydrofuran (5 mL). The reaction was then stirred at room temperature overnight. The mixture was quenched by the addition of saturated ammonium chloride solution (5 mL) at 0 ° C and then diluted with 80 mL of ethyl acetate. The organic phase was washed with water (60 mL) and brine (60 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/15) as eluent to obtain the desired product (1.0 g, 41% yield).

Step 3. Synthesis of 12-3

To a solution of 1-[4-[(4,4-difluorocyclohexyl)(2-methylprop-2-ene-1-yl)amino]-3-nitrophenyl]cyclopentane-1-carbonitrile (1.0 g, 2.48 mmol) in acetic acid (10 mL) was added iron (690 mg, 12.32 mmol). The mixture was then stirred at room temperature for 30 minutes. Ethyl acetate (60 mL) was added and the solid was filtered out. The reaction mixture was washed with saturated aqueous sodium bicarbonate solution (30 mL × 3), water (30 mL) and brine (30 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/8) as eluent to obtain the desired product (0.8 g, 86% yield).

Step 4. Synthesis of 12-4

To 1-[3-amino-4-[(4,4-difluorocyclohexyl)(2-methylprop-2-ene-1-yl)amino]phenyl]cyclopentane-1-carbonitrile (280mg, 0.75mmol) in ethyl acetate (3mL) solution, add palladium carbon (28mg) and triethylamine (0.3mL).Then reactant was stirred at room temperature for 30 minutes under hydrogen balloon.The mixture was filtered through diatomite and the filtrate was concentrated in vacuo.With ethyl acetate/petroleum ether (1/8) by silica gel column purification residue, obtain desired product (0.16g, 57% yield).

Step 5. Synthesis of 12-5

To a solution of 1-[3-amino-4-[(4,4-difluorocyclohexyl)(2-methylpropyl)amino]phenyl]cyclopentane-1-carbonitrile (160 mg, 0.43 mmol) in ethanol (4 mL) and water (1 mL) was added potassium hydroxide (1.12 g, 19.96 mmol). The mixture was then stirred at 100 ° C for 2 days. The mixture was cooled to room temperature and diluted with water (10 mL), extracted with ethyl acetate (10 mL×2), and washed with brine (10 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give the desired product (90 mg, 54% yield).

Step 6. Synthesis of 12

A solution of 1-[3-amino-4-[(4,4-difluorocyclohexyl)(2-methylpropyl)amino]phenyl]cyclopentane-1-carboxylic acid (90 mg, 0.23 mmol), 2,4-difluoro-1-isocyanatophenyl ester (53 mg, 0.34 mmol), and triethylamine (69 mg, 0.69 mmol) in tetrahydrofuran (3 mL) was stirred at room temperature for 0.5 hours. Ethyl acetate (20 mL) was then added, and the organic phase was washed with water (10 mL) and brine (10 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by preparative HPLC under the following conditions: [column: X Bridge Shield RP18 OBD column, 5 μm, 19×150 mm; mobile phase, water (10 MMOL/L NH ₄ HCO ₃ ) and ACN (ACN from 35.0% to 70.0% in 8 minutes); detector, 254 nm] to give the desired product (52.1 mg, 42% yield) as an off-white solid. LCMS(ES,m/z):550.50[M+H] ⁺ ； ¹ HNMR(300MHz,DMSO-d ₆ ,ppm): δ9.40(s,1H),8.24(s,1H),8.09(d,J＝2.1Hz,1H),7.96-7.88(m,1H),7.35-7.27( m,1H),7.15-6.94(m,3H),2.89-2.76(m,3H),2.08-1.24(m,17H),0.82(d,J=6.6Hz,6H).

Example 13

Step 1. Synthesis of 13-2

At 0 ℃, to a solution of lithium aluminum tetrahydride (3.0 g, 88.44 mmol) in tetrahydrofuran (30 mL) was added dropwise a solution of 4-(trifluoromethoxy)benzonitrile (5.0 g, 26.72 mmol) in tetrahydrofuran (30 mL). The reaction was then stirred at room temperature for 3 hours. The mixture was diluted with tetrahydrofuran (50 mL) at 0 ℃, and then water (3 mL), 15% sodium hydroxide (3 mL) and water (3 mL) were added to quench the mixture. The mixture was stirred at room temperature for 15 minutes. The mixture was filtered through diatomaceous earth, and the filtrate was dried over anhydrous magnesium sulfate and concentrated in vacuo to obtain the desired product (3.1 g, 61% yield).

Step 2. Synthesis of 13-3

At 0 ° C, HATU (9.25 g) was added in batches to a solution of 3,3,3-trifluoropropionic acid (2.28 g, 17.81 mmol) and N,N-diisopropylethylamine (4.2 g) in N,N-dimethylformamide (25 mL). The mixture was then stirred at 0 ° C for 5 minutes, followed by the addition of a solution of [4-(trifluoromethoxy)phenyl]methylamine (3.1 g, 16.22 mmol) in N,N-dimethylformamide (25 mL). The reaction was heated to room temperature and stirred for another 3 hours. Water (300 mL) was added, and the mixture was extracted with ethyl acetate (200 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/2) as eluent to give the desired product (2.4 g, 49% yield).

Step 3. Synthesis of 13-4

At 60 ° C, a solution of 3,3,3-trifluoro-N-[[4-(trifluoromethoxy)phenyl]methyl]propionamide (2.3 g, 7.64 mmol) in borane-tetrahydrofuran complex (10 mL, 1 M) was stirred for 1 hour. The reaction was quenched by adding methanol (10 mL) and concentrated HCl (3 mL) and stirred for another hour at 60 ° C. The mixture was concentrated in vacuo, then diluted with water (20 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/3) as eluent to obtain the desired product (1.0 g, 46% yield).

Step 4. Synthesis of 13-5

At 130 ° C, a solution of [4-(trifluoromethoxy)phenyl]methyl](3,3,3-trifluoropropyl)amine (720 mg, 2.51 mmol), 1-(4-fluoro-3-nitrophenyl)cyclopentane-1-carbonitrile (590 mg, 2.52 mmol) and N,N-diisopropylethylamine (0.65 g) in dimethyl sulfoxide (10 mL) was stirred overnight. The mixture was cooled to room temperature, diluted with ethyl acetate (60 mL), and washed with water (30 mL × 2) and brine (30 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/10) as eluent to give the desired product (0.47 g, 37% yield).

Step 5. Synthesis of 13-6

To a solution of 1-[3-nitro-4-([[4-(trifluoromethoxy)phenyl]methyl](3,3,3-trifluoropropyl)amino)phenyl]cyclopentane-1-carbonitrile (470 mg, 0.94 mmol) in acetic acid (5 mL) was added iron (0.26 g). The mixture was then stirred at room temperature for 0.5 hours. Ethyl acetate (30 mL) was added and the mixture was filtered through celite. The filtrate was concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/10) to give the desired product (0.33 g, 75% yield).

Step 6. Synthesis of 13-7

To a solution of 1-[3-amino-4-([[4-(trifluoromethoxy)phenyl]methyl](3,3,3-trifluoropropyl)amino)phenyl]cyclopentane-1-carbonitrile (330 mg, 0.70 mmol) in water (1 mL) and ethanol (4 mL) was added KOH (1.12 g, 19.96 mmol). The mixture was then stirred at 100 ° C overnight. The reactant was cooled to room temperature, diluted with water (10 mL), extracted with ethyl acetate (20 mL×2), and washed with brine (10 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give the desired product (160 mg, 47% yield).

Step 7. Synthesis of 13

To a solution of phenyl 2,4-difluoro-1-isocyanate (38 mg, 0.25 mmol) in tetrahydrofuran (3 mL) was added 1-[3-amino-4-([[4-(trifluoromethoxy)phenyl]methyl](3,3,3-trifluoropropyl)amino)phenyl]cyclopentane-1-carboxylic acid (80 mg, 0.16 mmol) and triethylamine (48 mg). The mixture was stirred at room temperature for 0.5 hours, then ethyl acetate (20 mL) was added. The resulting mixture was washed with water (10 mL) and brine (10 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by preparative HPLC under the following conditions: [column: X Bridge Prep C18 OBD column, 30×50 mm, 5 μm, 13 nm; mobile phase, water (0.1% FA) and ACN (ACN 60.0%-90.0% in 8 minutes, maintained at 90.0% in 2 minutes); detector, UV 254 nm] to give the desired product (47.4 mg, 45% yield) as an off-white solid. LCMS C ₃₀ H ₂₇ F ₈ N ₃ O ₄ (ES, m/z): 646.4[M+H] ⁺ ; ¹ HNMR (300MHz, DMSO-d ₆ ): δ12.23(brs,1H),9.34(s,1H),8.55(s,1H),8.11-8.01(m,2H),7.44(d,J＝8.7Hz,2H),7.37-7.27(m,3H),7.24-7.1 5(m,1H),7.08-7.02(m,1H),6.92-6.88(m,1H),4.18(s,2H),3.21-3.09(m,2H),2.73-2.50(m,4H),1.74-1.61(m,6H).

Example 14

Step 1. Synthesis of 14

A solution of 1-[3-amino-4-([[4-(trifluoromethoxy)phenyl]methyl](3,3,3-trifluoropropyl)amino)phenyl]cyclopentane-1-carboxylic acid (80 mg, 0.16 mmol), 1-isocyanato-4-methylbenzene (32 mg, 0.24 mmol), and triethylamine (48 mg) in tetrahydrofuran (3 mL) was stirred at room temperature for 0.5 hours. Ethyl acetate (20 mL) was added, and the resulting mixture was washed with water (10 mL) and brine (10 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by preparative HPLC under the following conditions: [column: X Bridge Prep C18 OBD column, 19×150 mm 5 um; mobile phase, water (0.1% FA) and ACN (ACN 50.0%-80.0% in 8 minutes, maintained at 80.0% in 2 minutes); detector, UV 254 nm] to give the desired product (31.5 mg, 32% yield) as an off-white solid. LCMS(ES,m/z):624.30[M+H] ⁺ ； ¹ HNMR(300MHz,DMSO-d ₆ ): δ9.39(s,1H),8.23(s,1H),8.18(d,J＝2.1Hz,1H),7.46-7.36(m,4H),7.26(d,J＝8.1Hz,2H),7.18-7.12(m ,3H),6.90-6.87(m,1H),4.17(s,2H),3.13-3.08(m,2H),2.50-2.45(m,4H),2.26(s,3H),1.75-1.62(m,6H).

Example 15

Step 1. Synthesis of 15-1

To a solution of 2-methylpropane-1-amine (480 mg, 6.56 mmol) in dichloromethane (10 mL) was added tetrahydropyran-4-one (723 mg, 7.22 mmol) and acetic acid (0.05 mL). The resulting mixture was stirred at room temperature for 0.5 hours, and then sodium cyanoborohydride (1.66 g, 26.42 mmol) was added. After stirring at room temperature for another 3 hours, the mixture was quenched by adding ammonium chloride solution (50 mL) and extracted with dichloromethane (50 mL × 2). The organic phase was washed with salt water (50 mL × 2), dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain the desired product (0.80 g, 78% yield).

Step 2. Synthesis of 15-2

At room temperature, to a solution of 1-(4-fluoro-3-nitrophenyl)cyclopentane-1-carbonitrile (1.0 g, 4.274 mmol) in dimethyl sulfoxide (30 mL) was added N,N-diisopropylethylamine (1.1 g, 8.548 mmol) and N-(2-methylpropyl)tetrahydropyran-4-amine (1.0 g, 6.411 mmol). The resulting solution was stirred at 130 ° C overnight. Water (30 mL) was added to quench the reaction and the mixture was extracted with ethyl acetate (30 mL×3). The combined organic layer was washed with brine (40×3), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:25) as eluent to give the desired product (0.30 g, 19% yield).

Step 3. Synthesis of 15-3

Under N ₂ , to a solution of 1- [4- [(2-methylpropyl) (tetrahydropyran-4-yl) amino] -3-nitrophenyl] cyclopentane -1- carbonitrile (0.3 g, 0.808 mmol) in methanol (10 mL) was added palladium on carbon. The suspension was degassed in vacuo and purged with H ₂ several times. The resulting mixture was stirred overnight at 25 ° C under a H ₂ balloon. The solid was filtered off and the filter cake was washed with methanol (10 mL × 3). The filtrate was concentrated in vacuo and applied to a silica gel column with ethyl acetate / petroleum ether (2: 7) as eluent to give the desired product (0.18 g, 65% yield).

Step 4. Synthesis of 15-4

To a solution of 1-[3-amino-4-[(2-methylpropyl)(tetrahydropyran-4-yl)amino]phenyl]cyclopentane-1-carbonitrile (0.18 g, 0.528 mmol) in ethanol (10 mL) was added potassium hydroxide (0.29 g, 5.28 mmol) and water (6 mL). The resulting solution was stirred at 130 ° C for 3 days. The reaction mixture was cooled to room temperature with water/ice bath. The pH value of the solution was adjusted to 5 with hydrochloric acid (1 N), and the mixture was extracted with ethyl acetate (20 mL×3). The combined organic layers were washed with brine (30 mL×3), dried over anhydrous sodium sulfate, and concentrated in vacuo to give the desired product (0.16 g, 84% yield).

Step 5. Synthesis of 15

Under _N2 , N,N-diisopropylethylamine (0.21 g, 1.663 mmol) and triphosgene (0.12 mg, 0.41 mmol) were added to a solution of 3-methyl-1,2-oxazol-5-amine (0.12 g, 1.22 mmol) in dichloromethane (12 mL). The resulting solution was stirred at room temperature for 20 minutes, followed by the addition of 1-[3-amino-4-[(2-methylpropyl)(tetrahydropyran-4-yl)amino]phenyl]cyclopentane-1-carboxylic acid (0.11 g, 0.31 mmol, 1.00 equiv), followed by the addition of triethylamine (0.19 g, 1.83 mmol, 6.00 equiv). The reaction was stirred at room temperature for another 2 hours. Methanol (8 mL) and water (20 mL) were added to quench the reaction, and the mixture was extracted with dichloromethane (20 mL × 3). The combined organic layers were washed with brine (30 mL x 3), dried over anhydrous sodium sulfate, and concentrated in vacuo. The crude product was purified by preparative HPLC [column, Xbridge, RP18, 19 x 150 mm; mobile phase, A: formic acid (aqueous solution) (0.1%), B: acetonitrile (35%-75% in 8 minutes); rate, 25 mL/min; detector, 254 nm] to give the product (72.3 mg, 49% yield) as a white solid. LCMS:(ES,m/z):[M+H]+485.4. ¹ HNMR(300MHz,CD ₃ OD,ppm)δ8.25(d,J＝2.1Hz,1H),7.22(d,J＝8.4Hz,1H),7.09(dd,J＝2.1Hz,J＝8.4Hz,1H),6.06(s,1H),3.94-3.89(m,2H),3.31(s,2H),2.9 2-2.83(m,3H),2.65-2.61(m,2H),2.24(s,3H),1.93-1.87(m,2H),1.78(s,6H),1.76-1.57(m,2H),1.48-1.32(m,1H),0.85(d,J＝6Hz,6H).

Example 16

Step 1. Synthesis of 16-1

At 0 ° C, sodium hydride (410 mg, 10.25 mmol) was added in batches to a solution of 2-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]acetonitrile (1.0 g, 3.46 mmol) in tetrahydrofuran (10 mL). At the same temperature, the mixture was stirred for 30 minutes, and then a solution of 1,3-dibromopropane (840 mg, 4.16 mmol) in THF (2 mL) was added. The resulting mixture was stirred for another 3 hours at room temperature. Water (2 mL) was then added to quench the reaction. The mixture was diluted with ethyl acetate (60 mL) and washed with water (30 mL) and brine (30 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/10) as eluent to obtain the desired product (0.4 g, 35% yield).

Step 2. Synthesis of 16-2

To 1-[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]cyclobutane-1-carbonitrile (400mg, 1.21mmol) in acetic acid (10mL) solution, add iron (680mg, 12.14mmol). The resulting mixture was stirred at room temperature for 1 hour. The reactant was filtered through diatomite and the filtrate was diluted with ethyl acetate (50mL), and washed with saturated aqueous sodium carbonate solution (20mL) and water (20mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. Ethyl acetate/petroleum ether (1/10) was used as eluent by silica gel column purification residue to obtain the desired product (0.198g, 54% yield).

Step 3. Synthesis of 16-3

To a solution of 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]cyclobutane-1-carbonitrile (198 mg, 0.66 mmol) in ethanol (4 mL) and water (2 mL) was added potassium hydroxide (110 mg, 1.96 mmol) at room temperature. The reaction was then stirred at 100 ° C overnight. The mixture was cooled to room temperature, diluted with water (20 mL) and extracted with ethyl acetate (20 mL×3). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with dichloromethane/methanol (10/1) as eluent to give the desired product (110 mg, 52% yield).

Step 4. Synthesis of 16

To a solution of 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]cyclobutane-1-carboxylic acid (82 mg, 0.26 mmol) in tetrahydrofuran (3 mL) were added triethylamine (53 mg, 0.52 mmol) and 2,4-difluoro-1-isocyanatophenyl ester (60 mg, 0.39 mmol). The reaction was then stirred at room temperature for 1 hour. The mixture was diluted with ethyl acetate (20 mL) and washed with water (10 mL x 2). The organic phase was concentrated in vacuo. The residue was purified by preparative HPLC using the following conditions: [column, X BridgeShield RP18 OBD column, 5 μm, 19 x 150 mm; mobile phase, water (10 mmol/L _{NH₄HCO₃ )} and ACN (ACN 15.0% to 65.0% over 8 minutes); detector, UV 254; 220 nm] to give the desired product (44.7 mg, 37% yield). LCMS(ES,m/z):474.50[M+H] ⁺ ; ¹ HNMR: (300MHz, DMSO-D ₆ ,ppm): δ9.23(s,1H),8.01-7.88(m,2H),7.77(s,1H),7.31-7.25(m,1H),7.08-7.00(m,2H),6.86( d,J＝6.9Hz,1H),2.90-2.61(m,6H),2.30-2.20(m,2H),1.82-1.62(m,4H),0.83(d,J＝6.6Hz,12H).

Example 17

Step 1. Synthesis of 17

To a solution of 3-methyl-1,2-oxazol-5-amine (98.1 mg, 1.00 mmol) and N,N-diisopropylethylamine (175 mg, 1.35 mmol) in tetrahydrofuran (3 mL) was added a solution of bis(trichloromethyl) carbonate (101 mg, 0.34 mmol) in THF (3 mL) at room temperature. The reaction was stirred for 15 minutes. Triethylamine (152 mg, 1.50 mmol) and 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]cyclobutane-1-carboxylic acid (80 mg, 0.25 mmol) were added, and the resulting mixture was stirred at room temperature for an additional 2 hours. The reaction was concentrated in vacuo. The residue was dissolved in methanol (4 mL) and purified by preparative HPLC under the following conditions: [column: X bridge, C18, 19×50 mm; mobile phase, H ₂ O (0.05% NH ₄ HCO ₃ )/MeCN, 35%-55% in 8 minutes; rate: 25 mL/min; detector, 254 nm] to give the desired product (27.5 mg, 6% yield) as a white solid. LCMS(ES,m/z):443.5[M+H] ⁺ ; ¹ HNMR: (300MHz, DMSO-d ₆ ,ppm): δ8.26(s,1H),7.88(d,J＝2.1Hz,1H),7.17(d,J＝8.3Hz,1H),6.91(dd,J＝8.2,2.2Hz,1H),5.99(s,1H),2.75-2.60( m,6H),2.40-2.25(m,2H),2.16(s,3H),1.91-1.80(m,1H),1.77-1.70(m,1H),1.68-1.55(m,2H),0.82(d,J＝6.5Hz,12H).

Example 18

Step 1. Synthesis of 18

At room temperature, to a solution of pyrimidine-5-amine (95.1 mg, 1.00 mmol) and N,N-diisopropylethylamine (175 mg, 1.35 mmol) in tetrahydrofuran (3 mL) was added dropwise a solution of di(trichloromethyl) carbonate (101 mg, 0.34 mmol) in THF (3 mL). After stirring for 15 minutes, triethylamine (152 mg, 1.50 mmol) and 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]cyclobutane-1-carboxylic acid (80 mg, 0.25 mmol) were added. The resulting mixture was stirred at room temperature for another 2 hours. The reactants were then concentrated in vacuo. The residue was purified by preparative HPLC under the following conditions: [column: X bridge, C18, 19×50 mm; mobile phase, H ₂ O (0.05% NH ₄ HCO ₃ )/MeCN, 35%-55% in 8 minutes; rate: 25 mL/min; detector, 254 nm] to give the desired product (72.7 mg, 17% yield) as a white solid. LCMS(ES,m/z):440.5[M+H] ⁺ . ¹ HNMR(300MHz,DMSO-d ₆ , ppm): δ10.01(s,1H),8.93(s,2H),8.81(s,1H),8.20(s,1H),7.90(d,J＝2.1Hz,1H),7.19(d,J＝8.3Hz,1H),6. 92(dd,J＝8.3,2.2Hz,1H),2.68(d,J＝6.9Hz,6H),2.42-2.26(m,2H),1.94-1.50(m,4H),0.86(d,J＝6.5Hz,12H).

Example 19

Step 1. Synthesis of 19-1

To a solution of 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]cyclobutane-1-carbonitrile (270mg, 0.90mmol) in tetrahydrofuran (5mL) was added 2,4-difluoro-1-phenylisocyanate (210mg, 1.35mmol) and triethylamine (182mg, 1.80mmol). The mixture was then stirred at room temperature for 2 hours. The reactant was diluted with ethyl acetate (20mL) and washed with water (10mL) and brine (10mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column purification using ethyl acetate/petroleum ether (1/1) as eluent to obtain the desired product (230mg, 56% yield).

Step 2. Synthesis of 19

At 90 ° C, a solution of 3- [2- [bis (2-methylpropyl) amino] -5- (1-cyanocyclobutyl) phenyl] -1- (2,4-difluorophenyl) urea (120 mg, 0.26 mmol), trimethylsilyl azide (152 mg, 1.32 mmol) and tetrabutylammonium fluoride (345 mg, 1.32 mmol) was stirred for 1 hour. The mixture was cooled to room temperature, diluted with ethyl acetate (10 mL), and washed with water (10 mL × 2). The organic phase was concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate / petroleum ether (5/1) as eluent to give the desired product (43.4 mg, 33% yield) as a white solid. LCMS(ES,m/z):498.5[M+H] ⁺ . ¹ HNMR:(300MHz,DMSO-d ₆ ,ppm): δ9.28(s,1H),8.03(s,1H),7.82-7.81(m,1H),7.80(s,1H),7.29(m,2H),7.14-7.12(m,1H),6.8 7(m,1H),2.81-2.79(m,2H),2.65-2.62(m,6H),1.89(m,2H),1.63-1.61(m,2H),0.83(d,J＝6.6Hz,12H).

Example 20

Step 1. Synthesis of 20-2

At 0 ° C, sodium hydride (10.66 g, 444.17 mmol) was added in batches to a solution of 2-(4-fluorophenyl)acetonitrile (20 g, 148.00 mmol) in tetrahydrofuran (200 mL). The mixture was stirred at 0 ° C for 30 minutes, followed by the addition of 1,3-dibromopropane (32.58 g, 161.38 mmol). The resulting mixture was stirred at room temperature overnight. The reaction was quenched with saturated ammonium chloride (50 mL) and extracted with ethyl acetate (300 mL × 3). The organic phase was washed with brine (300 mL × 2), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate / petroleum ether (1/20) as eluent to give the desired product (13.6 g, 52% yield).

Step 2. Synthesis of 20-3

At 0 ℃, potassium nitrate (11.6 g, 114.7 mmol) was added in batches to a solution of 1-(4-fluorophenyl)cyclobutane-1-carbonitrile (13.6 g, 77.6 mmol) in sulfuric acid (136 mL). The reaction was then stirred at room temperature overnight. The reaction was quenched with water (500 mL) and extracted with ethyl acetate (300 mL × 3). The organic phase was washed with brine (200 mL × 2), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column purification using ethyl acetate/petroleum ether (2/3) as eluent to obtain the desired product (12.6 g, 39% yield).

Step 3. Synthesis of 20-4

To a solution of 1-(4-fluoro-3-nitrophenyl)cyclobutane-1-carboxamide (12.6 g, 52.89 mmol) in 1,4-dioxane (126 mL) was added trifluoroacetic anhydride (16 mL) and triethylamine (6.7 mL). The resulting mixture was then heated to 100 ° C overnight. The reactants were cooled to room temperature, diluted with water (100 mL), and extracted with ethyl acetate (160 mL × 3). The organic phase was washed with brine (100 mL × 5), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/9) as eluent to obtain the desired product (10.5 g, 90% yield).

Step 4. Synthesis of 20-6

To a solution of 1-(4-fluoro-3-nitrophenyl)cyclobutane-1-carbonitrile (600 mg, 2.72 mmol) and N-ethylcyclohexylamine (416 mg, 3.27 mmol) in dimethyl sulfoxide (6 mL) was added N,N-diisopropylethylamine (1057.1 mg, 8.18 mmol). The resulting mixture was stirred at 100 ° C overnight. The reactants were cooled to room temperature, diluted with ethyl acetate (100 mL), and washed with water (100 mL×2) and brine (100 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by Flash-Prep-HPLC under the following conditions (IntelFlash-1): [column: silica gel column; mobile phase: methanol/dichloromethane increased from 0% to 8% in 20 minutes; detector, UV 254 nm] to give the desired product (700 mg, 78% yield).

Step 5. Synthesis of 20-7

To a solution of 1-[4-[cyclohexyl(ethyl)amino]-3-nitrophenyl]cyclobutane-1-carbonitrile (700 mg, 2.14 mmol) in acetic acid (7 mL) was added iron (2.39 g, 42.79 mmol). The mixture was stirred at room temperature for 30 minutes, then water (100 mL) was added. The pH value of the mixture was adjusted to 9 with aqueous sodium carbonate. The solid was filtered off, and the filtrate was extracted with ethyl acetate (100 mL × 3) and washed with brine (200 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give the desired product (600 mg, 94% yield).

Step 6. Synthesis of 20-8

To a solution of 1-[3-amino-4-[cyclohexyl(ethyl)amino]phenyl]cyclobutane-1-carbonitrile (550mg, 1.85mmol) in ethanol (9mL) and water (3mL) was added potassium hydroxide (1.56g, 27.76mmol). The resulting mixture was stirred at 100°C for 23 hours. The reactant was cooled to room temperature and diluted with water (100mL). The pH value of the mixture was adjusted to 4 with hydrogen chloride (1N), then extracted with ethyl acetate (100mL×3). The organic phase was washed with brine (100mL), dried over anhydrous sodium sulfate, and concentrated in vacuo to give the desired product (450mg, 77% yield).

Step 7. Synthesis of 20

To a solution of 1-[3-amino-4-[cyclohexyl(ethyl)amino]phenyl]cyclobutane-1-carboxylic acid (210 mg, 0.66 mmol) and 2,4-difluoro-1-isocyanatophenyl ester (154.7 mg, 1.00 mmol) in tetrahydrofuran (5 mL) was added triethylamine (200.9 mg, 1.99 mmol). The reaction was stirred at room temperature for 2.5 hours, after which ethyl acetate (50 mL) was added. The mixture was washed with water (50 mL x 2) and brine (50 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by preparative HPLC under the following conditions: [column, X Bridge Prep C18 OBD column, 19×150 mm 5 μm; mobile phase, water (10 mmol/L NH ₄ HCO ₃ )/CH ₃ CN; MeCN 25.0%-55.0% in 8 minutes; detector, UV 245 nm] to give the desired product (80.4 mg, 26% yield) as a white solid. LCMS(ES,m/z):472.5[M+H] ⁺ ; ¹ HNMR: (300MHz, DMSO-d ₆ ,ppm): δ9.39(s,1H),8.74(s,1H),8.19-7.94(m,2H),7.33-7.26(m,1H),7.22-6.97(m,2H),6.91-6.81(m,1H),3.00 (q,J＝7.0Hz,2H),2.75-2.60(m,3H),2.42-2.25(m,2H),1.97-1.53(m,7H),1.20-1.00(m,5H),0.82(t,J＝7.0Hz,3H).

Example 21

Step 1. Synthesis of 21-1

To a solution of 1-(4-fluoro-3-nitrophenyl)cyclobutane-1-carbonitrile (1 g, 4.54 mmol) in dimethyl sulfoxide (10 mL) was added N,N-diisopropylethylamine (1.76 g, 13.62 mmol) followed by N-(2-methylpropyl)cyclohexylamine (777 mg, 5.00 mmol). The mixture was stirred at 100° C. for 16 hours. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was purified by Flash-Prep-HPLC using the following conditions: [column: C18 silica gel; mobile phase A: water (0.05% TFA)), mobile phase B: ACN; gradient: 45%-100% ACN; detector: UV 254 nm] to give the desired product (0.8 g, 50% yield).

Step 2. Synthesis of 21-2

To a solution of 1-[4-[cyclohexyl(2-methylpropyl)amino]-3-nitrophenyl]cyclobutane-1-carbonitrile (800mg, 2.25mmol) in ethyl acetate (5mL) and acetic acid (5mL) was added iron (1.26g, 22.56mmol). The reaction was then stirred at room temperature for 0.5 hour. The mixture was diluted with ethyl acetate (1500mL) and the solid was filtered out. The pH value of the filtrate was adjusted to 9 with sodium carbonate. The organic phase was washed with brine (50mL×2), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column purification using ethyl acetate/petroleum ether (1/6) as eluent to obtain the desired product (0.5g, 68% yield).

Step 3. Synthesis of 21-3

To a solution of 1-[3-amino-4-[cyclohexyl(2-methylpropyl)amino]phenyl]cyclobutane-1-carbonitrile (300mg, 0.92mmol)) in ethanol (6mL) and water (1.5mL) was added potassium hydroxide (900mg, 16.04mmol). The reaction was then stirred at 95°C for 16 hours. After cooling to room temperature, the mixture was concentrated in vacuo and the residue was dissolved in water (50mL). The pH value of the mixture was adjusted to 4 with hydrogen chloride (1N). The mixture was extracted with ethyl acetate (50mL×2) and washed with saturated brine (50mL×2). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give the desired product (0.3g, 94% yield).

Step 4. Synthesis of 21

To a solution of 1-[3-amino-4-[cyclohexyl(2-methylpropyl)amino]phenyl]cyclobutane-1-carboxylic acid (300 mg, 0.87 mmol) in tetrahydrofuran (6 mL) was added triethylamine (264 mg, 2.61 mmol) and 2,4-difluoro-1-isocyanatophenyl ester (149 mg, 0.96 mmol). The reaction was then stirred at room temperature for 1.5 hours. The mixture was concentrated in vacuo and the residue was purified by preparative HPLC under the following conditions: [column: Waters X-bridge C18, 5 μm, 19×150 mm; mobile phase A: water (0.05% NH ₄ HCO ₃ ), mobile phase B: ACN; gradient: 25% ACN to 50% ACN over 8 minutes; detector: UV 254 nm] to give the desired product (62.5 mg, 14% yield) as a white solid. LCMS:(ES,m/z):500.30[M+H] ⁺ . ¹ H NMR(300MHz,DMSO-d ₆ ,ppm): δ9.31(s,1H),8.09(s,1H),7.87-7.79(m,2H),7.27-7.21(m,1H),7.05-6.95(m,2H),6.81-6.78(m,1H),2.8 3-2.52(m,4H),2.28-2.12(m,2H),1.93-1.53(m,6H),1.52-1.38(m,1H),1.37-0.86(m,6H),0.75(d,J＝6.5Hz,6H).

Example 22

Step 1. Synthesis of 22-1

To a solution of 1-(4-fluoro-3-nitrophenyl)cyclobutane-1-carbonitrile (1 g, 4.54 mmol) in dimethyl sulfoxide (10 mL) was added N-(2-methoxyethyl)cyclohexylamine (1.16 g, 7.38 mmol) and N,N-diisopropylethylamine (1.74 g). The mixture was then stirred at 100 ° C overnight. The reactants were cooled to room temperature, quenched with water (50 mL), and extracted with ethyl acetate (300 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give the desired product (0.7 g, 43% yield).

Compound 22 was synthesized according to steps 2-4 similar to those in Example 21.

Example 22: LCMS (ES, m/z): 502.4 [M+H] ⁺ ; ¹ HNMR (300 MHz, DMSO-d ₆ ): δ9.34(s,1H),8.61(s,1H),8.06(d,J＝1.8Hz,1H),8.01-7.93(m,1H),7.33-7.26(m,1H),7.16(d,J＝8.1Hz,1H),7.06-7.00(m,1H), 6.87-6.84(m,1H),3.24-3.07(m,7H),2.78-2.62(m,3H),2.31-2.26(m,2H),1.89-1.66(m,6H),1.53-1.49(m,1H),1.23-1.00(m,4H).

Example 23

Step 1. Synthesis of 23-1

To a solution of 1-(4-fluoro-3-nitrophenyl)cyclobutane-1-carbonitrile (500 mg, 2.27 mmol) in dimethyl sulfoxide (10 mL) was added 2-methyl-1-[(2-methylpropyl)amino]propane-2-ol (330 mg, 2.27 mmol) and N,N-diisopropylethylamine (354 mg, 2.72 mmol). The resulting mixture was stirred at 100 ° C for 12 hours. After cooling to room temperature, the mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL×3). The organic matter was washed with brine (50 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo to give the desired product (350 mg, 45%).

Compound 23 was synthesized according to steps 2-4 similar to those in Example 21.

Example 23: LCMS (ES, m/z): 490.3 [M+H] ⁺ ; ¹ HNMR (300MHz, CD ₃ OD,ppm): δ7.98(s,1H),7.93-7.85(m,1H),7.25(d,J＝8.4Hz,1H),7.05-6.90(m,3H),3.02(s,2H),2.87(d,J＝6.9Hz,2H),2.82 -2.74(m,2H),2.52-2.42(m,2H),2.01-1.92(m,1H),1.92-1.83(m,1H),1.60-1.55(m,1H),1.14(s,6H),0.87(d,J＝6.6Hz,6H).

Example 24

Step 1. Synthesis of 24-1

To a solution of 1-(4-fluoro-3-nitrophenyl)cyclobutane-1-carbonitrile (500 mg, 2.27 mmol) in dimethyl sulfoxide (6 mL) was added N-ethyltetrahydropyran-4-amine (350 mg, 2.71 mmol) and N,N-diisopropylethylamine (870 mg). The resulting mixture was then stirred at 100 ° C overnight. The reactants were cooled to room temperature, diluted with water (10 mL), and extracted with ethyl acetate (30 mL × 3). The organic phase was washed with brine (30 mL × 5), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (3/2) as eluent to obtain the desired product (480 mg, 64% yield).

Compound 24 was synthesized according to similar steps 2-4 in Example 21.

Example 24: LCMS (ES, m/z): 474.3 [M+H] ⁺ ; ¹ H NMR (300 MHz, DMSO-d ₆ , ppm): δ 9.42 (s, 1H), 8.80 (s, 1H), 8.14-8.13 (d, J=2.1 Hz, 1H), 8.07-7.99 (m, 1H), 7.34-7.26 (m, 1H), 7.17 (d, J=8.2 Hz, 1H), 7.06-7.00 (m, 1H), 6.87 (dd, J=8.1, 2.1 Hz, 1H),3.84-3.81(m,2H),3.27-3.23(m,2H),3.19-2.95(m,3H),2.72-2.63(d,J＝4.5H z,2H),2.39-2.29(m,2H),1.97-1.69(m,4H),1.42-1.37(m,2H),0.84-0.79(m,3H).

Example 25

Step 1. Synthesis of 25-1

At 100 ° C, a solution of 1-(4-fluoro-3-nitrophenyl)cyclobutane-1-carbonitrile (1 g, 4.54 mmol), N-(2-methylpropyl)tetrahydropyran-4-amine (1.43 g, 9.09 mmol) and N,N-diisopropylethylamine (2.34 g, 18.11 mmol) in DMSO (20 mL) was stirred for 16 hours. The mixture was then cooled to room temperature, diluted with water (200 mL), and extracted with ethyl acetate (100 mL × 2). The organic phase was washed with water (100 mL × 5) and brine (30 mL × 2), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/10) as eluent to give the desired product (0.4 g, 19% yield).

Compound 25 was synthesized according to similar steps 2-4 in Example 21.

Example 25: LCMS (ES, m/z): 502.4 [M+H] ⁺ ; ¹ H NMR: (300 MHz, DMSO-d ₆ , ppm): δ11.8 (brs, 1H), 9.42 (s, 1H), 8.38 (s, 1H), 8.02 (d, J=1.8, 1H), 8.00-7.82 (m, 1H), 7.34-7.30 (m, 1H), 7.29-7.16 (m, 1H), 7.10-6.90 (m, 1H), 6.93-6.80 (m, 1H),3.85-3.81(m,2H),3.25-3.15(m,2H),2.95-2.59(m,5H),2.42-2.12(m,2H) ,2.00-1.62(m,4H),1.61-1.37(m,2H),1.36-1.17(m,1H),0.82(d,J＝6.6Hz,6H).

Example 26

Step 1. Synthesis of 26-1

To a solution of tetrahydrothiopyran-4-one (4.77 g, 41.06 mmol) and 2-methylpropane-1-amine (2 g, 27.35 mmol) in dichloromethane (60 mL) was added acetic acid (0.1 mL). The reaction was then stirred at room temperature for 0.5 hours. Sodium cyanoborohydride (6.87 g, 109.33 mmol) was added and the reaction was then stirred at room temperature for 16 hours. The mixture was diluted with ethyl acetate (200 mL), washed with brine (30 mL × 2), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column purification using ethyl acetate/petroleum ether (1/7) as eluent to obtain the desired product (1.4 g, 30% yield).

Step 2. Synthesis of 26-2

To a solution of 1-(4-fluoro-3-nitrophenyl)cyclobutane-1-carbonitrile (1.04 g, 4.72 mmol) and N-(2-methylpropyl)tetrahydrothiopyran-4-amine (750 mg, 4.33 mmol) in dimethyl sulfoxide (10 mL) was added N,N-diisopropylethylamine (835 mg, 6.46 mmol). The reaction was then stirred at 100° C. for 2 days. The mixture was cooled to room temperature and diluted with water (100 mL). The mixture was extracted with ethyl acetate (100 mL×2) and washed with water (100 mL×3) and brine (100 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by Flash-Prep-HPLC under the following conditions: [column: C18 silica gel; mobile phase A: water (0.05% TFA), mobile phase B: ACN; gradient: 55% ACN-95% ACN; detector: UV 254 nm] to give the desired product (370 mg, 21% yield).

Step 3. Synthesis of 26-3

At 0 ℃, to 1-[4-[(2-methylpropyl) (tetrahydrothiopyran-4-yl) amino] -3-nitrophenyl] cyclobutane-1-carbonitrile (340mg, 0.91mmol) in dichloromethane (10mL) solution, add 3-chlorobenzene-1-carbon peroxy acid (240mg, 1.39mmol).Then the mixture was stirred at 0 ℃ for 0.5 hour, and at room temperature for another 1.5 hours. The solid was filtered off and the filtrate was concentrated in vacuo. With ethyl acetate/petroleum ether (1/2) as eluent, the desired product (380mg, crude product) was obtained by silica gel column purification residue.

Step 4. Synthesis of 26-4

To the mixture of 26-3 (330mg, 0.81mmol) in ethyl acetate (8mL) and methanol (8mL), nickel (200mg, 3.41mmol) is added. The suspension is vacuum degassed and purged three times with _H. The mixture is stirred at room temperature for 30 minutes under _H balloon. Solid is filtered off, and the filtrate is concentrated in vacuo to obtain the desired product (265mg, 87% yield).

Step 5. Synthesis of 26-5

To a solution of ethanol (6 mL) and water (1.5 mL) of 26-4 (265 mg, 0.71 mmol) was added sodium hydroxide (1.2 g, 30.00 mmol). The mixture was then stirred at 90 ° C for 16 hours. The reactant was cooled to room temperature and diluted with ethyl acetate (100 mL) and water (100 mL). The pH value of the mixture was adjusted to 4 with hydrogen chloride (1 N). The mixture was extracted with ethyl acetate (100 mL × 2) and washed with brine (100 mL × 2). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to obtain the desired product (180 mg, 65% yield).

Step 6. Synthesis of 26

To a solution of 26-4 (180 mg, 0.46 mmol) in tetrahydrofuran (5 mL) were added 2,4-difluoro-1-phenylisocyanate (78 mg, 0.50 mmol) and triethylamine (69 mg, 0.68 mmol) in sequence. The mixture was then stirred at room temperature for 2 hours. The mixture was concentrated in vacuo, and the residue was purified by preparative HPLC under the following conditions: [column: X bridge, C18, 5 μm, 19×150 mm; mobile phase A: water (0.05% NH ₄ HCO ₃ ), mobile phase B: ACN; gradient: 35% ACN to 60% ACN over 8 minutes; detector: UV 254 nm] to give the desired product (92.2 mg, 37% yield) as a white solid. LCMS: (ES, m/z): 550.1 [M+H] ⁺ . ¹ H-NMR: (300 MHz, DMSO-d ₆ ,ppm): δ9.44(s,1H),8.21(s,1H),8.05(s,1H),7.99-7.91(m,1H),7.35-7. 27(m,1H),7.16(d,J＝8.1Hz,1H),7.11-6.98(m,1H),6.89-6.85(m,1H),3.25 -3.10(m,2H),3.09-2.92(m,4H),2.84-2.60(m,4H),2.39-2.19(m,4H),2.0 2-1.82(m,2H),1.81-1.68(m,1H),1.41-1.21(m,1H),0.83(d,J＝6.6Hz,6H).

Example 27

Step 1. Synthesis of 27-1

To a solution of 1-[4-[(2-methylpropyl)(tetrahydropyran-4-yl)amino]-3-nitrophenyl]cyclobutane-1-carbonitrile (250 mg, 0.70 mmol) in acetic acid (2.5 mL) was added iron (392 mg). The resulting solution was stirred at room temperature for 0.5 hours. The reactant was diluted with ethyl acetate (50 mL) and water (50 mL), and the pH value of the mixture was adjusted to 9 with aqueous sodium carbonate. The solid was filtered off and the filtrate was extracted with ethyl acetate (50 mL × 2). The organic phase was washed with brine (50 mL × 2) and water (60 mL × 2), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/2) as eluent to obtain the desired product (80 mg, 35% yield).

Step 2. Synthesis of 27-2

To a solution of 1-[3-amino-4-[(2-methylpropyl)(tetrahydropyran-4-yl)amino]phenyl]cyclobutane-1-carbonitrile (80 mg, 0.24 mmol) in ethanol (3 mL) and water (1 mL) was added potassium hydroxide (449 mg, 8.00 mmol). The resulting solution was stirred at 95 ° C for 16 hours. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was dissolved in water (50 mL) and the pH value of the solution was adjusted to 4 with hydrogen chloride (1 N). The resulting mixture was extracted with ethyl acetate (50 mL×2). The organic phase was washed with brine (50 mL×2), dried over anhydrous sodium sulfate, and concentrated in vacuo to give the desired product (70 mg, 83% yield).

Step 3. Synthesis of 27

To a solution of 3-methyl-1,2-oxazol-5-amine (57 mg, 0.58 mmol) in tetrahydrofuran (1.5 mL) was added N,N-diisopropylethylamine (101 mg, 0.78 mmol), followed by a solution of bis(trichloromethyl) carbonate (58 mg, 0.20 mmol) in tetrahydrofuran (1.5 mL). The resulting solution was stirred at room temperature for 10 minutes. A solution of 1-[3-amino-4-[(2-methylpropyl)(tetrahydropyran-4-yl)amino]phenyl]cyclobutane-1-carboxylic acid (70 mg, 0.20 mmol) in tetrahydrofuran (1.5 mL) and triethylamine (87 mg, 0.86 mmol) were added, and the reaction was stirred at room temperature for an additional hour. The resulting solution was then concentrated in vacuo and the residue was purified by Flash-Prep-HPLC under the following conditions [column: C18 silica gel; mobile phase A: ACN, mobile phase B: water (0.05% FA)/; gradient: 28% ACN-55% ACN over 8 minutes; detector, UV 254 nm] to give the desired product (10.5 mg) as a white solid. LCMS (ES, m/z): 471.3 [M+H] ⁺ . ¹ H NMR: (300 MHz, DMSO-d ₆ ,ppm): δ11.28(s,1H),8.51(s,1H),8.10(s,1H),8.23(d,J＝8.1Hz,1H), 6.92(dd,J＝2.1,8.1Hz,1H),6.00(s,1H),3.90-3.78(m,2H),3.24-3.16( m,2H),2.86-2.62(m,5H),2.43-2.30(m,2H),2.21-2.13(m,3H),2.00-2 .66(m,4H),1.60-1.47(m,2H),1.37-1.21(m,1H),0.82(d,J＝6.0Hz,6H).

Example 28

Step 1. Synthesis of 28-1

To a solution of 1-methylpiperidin-4-one (3.06 g, 27.07 mmol) and 2-methylpropane-1-amine (1.8 g, 24.61 mmol) in dichloromethane (40 mL) was added acetic acid (0.1 mL, catalyst) at 0°C. Sodium cyanoborohydride (6.2 g, 98.51 mmol, 4.00 equivalents) was added and the reaction was stirred at room temperature for 2.5 hours. The reaction mixture was diluted with ethyl acetate (400 mL) and washed with water (400 mL x 2) and brine (400 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was dissolved in methanol (3 mL) and a solution of oxalic acid dihydrate (2.2 g) in methanol (10 mL) was added. The solid was collected by filtration and redissolved in water (50 mL). The pH of the solution was adjusted to 9 with aqueous sodium hydroxide solution (15%). The resulting mixture was extracted with dichloromethane (400 mL x 4). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give the desired product (650 mg, 16% yield).

Step 2. Synthesis of 28-2

To a solution of 1-methyl-N-(2-methylpropyl)piperidin-4-amine (441.4 mg, 2.59 mmol) and 1-(4-fluoro-3-nitrophenyl)cyclobutane-1-carbonitrile (474 mg, 2.15 mmol) in dimethyl sulfoxide (6 mL) was added N,N-diisopropylethylamine (883.1 mg, 6.83 mmol). The reaction was then stirred at 100°C for 16 hours. The reaction mixture was cooled to room temperature and diluted with ethyl acetate (100 mL). The mixture was washed with water (100 mL x 2) and brine (100 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by Flash-Prep-HPLC under the following conditions: [column: silica gel; mobile phase A: dichloromethane, mobile phase B: methanol; gradient: 0% methanol to 8% methanol over 20 minutes; detector: UV 254 nm] to obtain the desired product (600 mg, 75% yield).

28 was synthesized according to similar steps 2-4 in Example 21.

Example 28: LRMS: (ES, m/z): 515.3[M+H] ⁺ . ¹ HNMR (300MHz, DMSO-d ₆ ,ppm): δ9.41(s,1H),8.31(s,1H),7.97-7.88(m,2H),7.34-7.26(m,1H),7.15(d,J＝8.1Hz,1H),7.07-7.04(m,1H),6.87(dd,J＝8 .1,2.4Hz,1H),2.77-2.63(m,5H),2.39-2.27(m,2H),2.08(s,3H),1.87-1.73(m,6H),1.52-1.21(m,5H),0.82(d,J＝6.6Hz,6H).

Example 29

Step 1. Synthesis of 29-2

At -78 ℃, to 2-(4-fluorophenyl) acetonitrile (1.35g, 9.99mmol) in tetrahydrofuran (10mL) solution, add MeLi (10mL, 1M) dropwise. The gained mixture is stirred 30 minutes at the same temperature, then 2-(bromomethyl) oxirane (1.37g, 10.00mmol) and methylmagnesium iodide (4mL) are added successively. The mixture obtained is warming up to room temperature and stirred for another 12 hours. Add water/ice (200mL) to quench the reaction, and extract with ethyl acetate (50mL×3). The organic phase is dried over anhydrous sodium sulfate and concentrated in vacuo. Use ethyl acetate/petroleum ether (1/20) as eluent by silica gel column purification residue, obtain desired product (1.3g, 68% yield).

Step 2. Synthesis of 29-3

At 0 ° C, to a solution of 1- (4- fluorophenyl) -3- hydroxycyclobutane -1- nitrile (750mg, 3.92mmol) in dichloromethane (10mL) was added Dess-Martin periodinane (2.5g, 0.01mmol) in batches. The resulting mixture was then stirred at room temperature for 12 hours and then quenched by adding water/ice (100mL). The solid was filtered off and washed with dichloromethane (50mL×2). The filtrate was then extracted with dichloromethane (50mL×2). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/20) as eluent to obtain the desired product (550mg, 74% yield).

Step 3. Synthesis of 29-4

At 0 ℃, diethylamino sulfur trifluoride (563mg, 3.49mmol) was added dropwise to a solution of 1-(4-fluorophenyl)-3-oxocyclobutane-1-carbonitrile (210mg, 1.11mmol) in dichloromethane (2mL). The mixture was then stirred at room temperature for 12 hours. Water/ice (20mL) was added to quench the reaction, extracted with ethyl acetate (10mL×3), and washed with brine (60mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. Ethyl acetate/petroleum ether (1/10) was used as eluent to pass through a silica gel column purification residue to obtain the desired product (160mg, 68% yield).

Step 4. Synthesis of 29-5

At 0 ° C, potassium nitrate (92 mg) was added in batches to a solution of 3,3-difluoro-1-(4-fluorophenyl)cyclobutane-1-carbonitrile (160 mg, 0.76 mmol) in concentrated sulfuric acid (2 mL). The mixture was then stirred at room temperature for 12 hours. Water/ice (20 mL) was added to quench the reaction, extracted with ethyl acetate (20 mL × 3), and washed with brine (60 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/30) as eluent to obtain the desired product (150 mg, 72% yield).

Step 5. Synthesis of 29-6

To a solution of 3,3-difluoro-1-(4-fluoro-3-nitrophenyl)cyclobutane-1-carboxamide (150 mg, 0.55 mmol) in 1,4-dioxane (5 mL) was added trifluoroacetic anhydride (0.5 mL) and triethylamine (1.1 mL). The resulting mixture was stirred at 120 ° C for 12 hours. The reactant was cooled to room temperature, water/ice (20 mL) was added to quench the reaction, extracted with ethyl acetate (20 mL × 3), and washed with brine (60 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/20) as eluent to obtain the desired product (100 mg, 71% yield).

Step 6. Synthesis of 29-7

To a solution of 3,3-difluoro-1-(4-fluoro-3-nitrophenyl)cyclobutane-1-carbonitrile (100 mg, 0.39 mmol, 1.00 equiv) and N,N-diisopropylethylamine (76 mg, 0.59 mmol) in dimethyl sulfoxide (2 mL) was added bis(2-methylpropyl)amine (60 mg, 0.46 mmol). The mixture was then stirred at 90 ° C for 12 hours. The reactants were cooled to room temperature, quenched with water/ice (50 mL), extracted with ethyl acetate (50 mL × 3), and washed with brine (20 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/20) as eluent to give the desired product (60 mg, 42% yield).

Step 7. Synthesis of 29-8

To a solution of 3,3-difluoro-1-(4-fluoro-3-nitrophenyl)cyclobutane-1-carbonitrile (100 mg, 0.39 mmol) and N,N-diisopropylethylamine (76 mg, 0.59 mmol) in dimethyl sulfoxide (2 mL) was added bis(2-methylpropyl)amine (60 mg, 0.46 mmol). The resulting mixture was then stirred at 90 ° C for 12 hours. The reactants were cooled to room temperature, quenched with water/ice (50 mL), extracted with ethyl acetate (50 mL × 3), and washed with brine (20 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/20) as eluent to obtain the desired product (60 mg, 42% yield).

Step 8. Synthesis of 29-9

To a solution of 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]-3,3-difluorocyclobutane-1-carbonitrile (40 mg, 0.12 mmol) in ethanol (2 mL) and water (1 mL) was added potassium hydroxide (10 mg, 0.18 mmol). The resulting mixture was then stirred at room temperature for 36 hours. Water (10 mL) was added and the mixture was extracted with ethyl acetate (10 mL × 2). The combined organic phases were washed with brine (10 mLx3), dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by Pre-TLC with ethyl acetate/petroleum ether (1/10) to give the desired product (40 mg, 88% yield).

Step 9. Synthesis of 29

To a solution of 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]-3,3-difluorocyclobutane-1-carboxylic acid (40 mg, 0.11 mmol) and triethylamine (17 mg, 0.17 mmol) in tetrahydrofuran (4 mL) was added 2,4-difluoro-1-phenylisocyanate (21 mg, 0.14 mmol). The resulting mixture was then stirred at room temperature for 12 hours. Water/ice (20 mL) was added to quench the reaction and extracted with ethyl acetate (10 mL × 3). The organic phase was washed with brine (30 mL × 2), dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by Pre-TLC to give the desired product (15.7 mg, 27% yield). LCMS(ES,m/z):510.5[M+H] ⁺ ; ¹ HNMR: (300MHz, DMSO-d ₆ ,ppm): δ13.01(brs,1H),9.31(s,1H),8.1(s,1H),7.97-7.85(m,2H),7.31(t,J＝6Hz,1H),7.17(d,J＝6Hz,1H),7.06(t, J＝6Hz,1H),6.93(t,J＝6Hz,1H),3.25(s,1H),3.00-2.86(m,3H),2.67(d,J＝6Hz,4H),1.70-1.61(m,2H),0.91(s,12H).

Example 30

Step 1. Synthesis of 30-1

To a solution of 1-[3-amino-4-[bis(2-methylpropyl)amino]phenyl]-3,3-difluorocyclobutane-1-carbonitrile (500mg, 1.49mmol) and triethylamine (196mg, 1.94mmol) in dichloromethane (5mL) was added 2,4-difluoro-1-phenylisocyanate (254mg, 1.64mmol). The resulting mixture was then stirred at room temperature for 12 hours. Water (50mL) was added to quench the reaction, extracted with ethyl acetate (30mL×3), and washed with brine (50mL×2). The organic layer was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/20) as eluent to obtain the desired product (700mg, 96% yield).

Step 2. Synthesis of 30

A solution of 1-[2-[bis(2-methylpropyl)amino]-5-(1-cyano-3,3-difluorocyclobutyl)phenyl]-3-(2,4-difluorophenyl)urea (500 g, 1.02 mol), trimethylsilyl azide (1.2 g, 10.42 mmol) and tetrabutylammonium fluoride (2.7 g, 10.33 mmol) was stirred at 85 ° C for 12 hours. The reactants were then cooled to room temperature and quenched with water (100 mL). The mixture was extracted with ethyl acetate (100 mL × 3) and washed with brine (50 mL × 2). The organic layer was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by preparative HPLC under the following conditions: [column: X Bridge Prep C18 OBD, 19x150nm 5um; mobile phase water (0.05% TFA) and ACN/MEOH (15%-60.0% in 8 minutes); detector, UV 254nm] to give the desired product (132.9mg, 24% yield) as a white solid. LCMS(ES,m/z):534.2[M+H] ⁺ . ¹ HNMR:(300MHz,DMSO-d ₆ , ppm): δ9.33(s,1H),8.06(s,1H),7.96-7.87(m,2H),7.31(t,J＝6Hz,1H),7.19(d,J＝8.4Hz,1H),7. 08-6.95(m,2H),3.44-3.35(m,4H),2.66(d,J＝6.9Hz,4H),1.68-1.59(m,2H),0.83(d,J＝6Hz,12H).

Example 31

Step 1. Synthesis of 31-2

A solution of 4-fluoro-3-nitrophenol (1g, 6.37mmol) and potassium carbonate (1.76g, 12.73mmol) in N,N-dimethylformamide (15mL) was cooled to 0°C. Methyl 2-bromoacetate (1.17g, 7.65mmol) was added dropwise. The mixture was stirred at room temperature overnight. Water (15mL) was then added to quench the reaction and extracted with ethyl acetate (50mL×3). The organic layer was washed with brine (50mL×3), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column using ethyl acetate/petroleum ether (1:10-1:5) as eluent to obtain the desired product (800mg, 55% yield).

Step 2. Synthesis of 31-3

At 60 ℃, 2-(4-fluoro-3-nitrophenoxy) methyl acetate (800mg, 3.49mmol), bis(2-methylpropyl) amine (676mg, 5.23mmol) and N-ethyl-N-isopropylpropane-2-amine (1.35g, 10.45mmol) in dimethyl sulfoxide (10mL) solution was stirred for 4 hours. The reactant was then cooled to room temperature and diluted with water (10mL). The mixture was extracted with ethyl acetate (50mL×3). The organic layer was washed with brine (50mL×3), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column purification using ethyl acetate/petroleum ether (1:20-1:5) as eluent to obtain the desired product (800mg, 68% yield).

Step 3. Synthesis of 31-4

To a solution of 2-[4-[bis(2-methylpropyl)amino]-3-nitrophenoxy]methyl acetate (800 mg, 2.36 mmol) in ethyl acetate (10 mL) and methanol (1 mL) was added palladium on carbon (500 mg). The mixture was stirred at room temperature for 2 hours under a hydrogen balloon. The solid was filtered off and washed with methanol (10 mL × 3). The filtrate was concentrated in vacuo and the residue was purified by silica gel column using ethyl acetate/petroleum ether (1:10 to 1:3) as eluent to obtain the desired product (400 mg, 55% yield).

Step 4. Synthesis of 31-5

To a solution of 2-[3-amino-4-[bis(2-methylpropyl)amino]phenoxy]methyl acetate (300 mg, 0.97 mmol) in tetrahydrofuran (5 mL) was added triethylamine (147 mg, 1.45 mmol) followed by 2,4-difluoro-1-phenylisocyanate (181 mg, 1.17 mmol). The mixture was stirred at room temperature for 2 hours and then concentrated in vacuo. The residue was purified by silica gel column using ethyl acetate/petroleum ether (1:10-1:3) as eluent to give the desired product (180 mg, 40% yield).

Step 5. Synthesis of 31

To a solution of methyl 2-[4-[bis(2-methylpropyl)amino]-3-[[(2,4-difluorophenyl)carbamoyl]amino]phenoxy]acetate (150 mg, 0.32 mmol) in tetrahydrofuran (5 mL) and methanol (1 mL) was added sodium hydroxide (0.3 mL, 15% aqueous solution). The reaction was stirred at room temperature for 2 hours. The mixture was concentrated in vacuo and the residue was purified by preparative HPLC using the following conditions: [column, Waters X-bridge RP18, 19×150 mm, 5 μm; mobile phase, ACN/water (0.05% TFA) 17%-43% in 7 minutes, flow rate: 20 mL/min; detector, 254 nm] to give the desired product (30.7 mg, 21% yield) as an off-white solid. LCMS (ES, m/z): 450.2[M+H] ⁺ ; 1HNMR: (300MHz, DMSO-d ₆ , ppm): δ12.90(s,1H),9.33(s,1H),8.23(s,1H),7.91-7.82(m,1H),7.59(d,J＝2.7Hz,1H),7.33-7.25(m,1H),7.12(d,J＝8.7Hz,1 H),7.08-7.02(m,1H),6.51(dd,J＝8.7,2.7Hz,1H),4.54(s,2H),2.61(d,J＝6.9Hz,4H),1.64-1.55(m,2H),0.83(d,J＝6.6Hz,12H).

Example 32

Step 1. Synthesis of 32-1

To a solution of 4-fluoro-3-nitrophenol (10 g, 63.65 mmol) and potassium carbonate (17 g, 123.00 mmol) in N,N-dimethylformamide (40 mL) was added 2-bromo-2-methylpropionic acid methyl ester (23 g, 127.05 mmol). After stirring at 60 ° C for 2.5 hours, water (150 mL) was added to quench the reaction and the mixture was extracted with ethyl acetate. The organic phase was washed with salt water, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column purification using ethyl acetate/petroleum ether (1/50) as eluent to obtain the desired product (11 g, 67% yield).

Step 2. Synthesis of 32-2

A solution of methyl 2-(4-fluoro-3-nitrophenoxy)-2-methylpropanoate (10 g, 38.88 mmol), diisopropylethylamine (10 g, 77.38 mmol) and bis(2-methylpropyl)amine (7 g, 54.16 mmol) in dimethyl sulfoxide (40 mL) was stirred overnight at 100°C. After cooling to room temperature, the reaction was quenched with water (100 mL) and extracted with ethyl acetate. The organic phase was washed with water and brine, dried over anhydrous sodium sulfate, and concentrated in vacuo to give the desired product (11 g, 77% yield).

Step 3. Synthesis of 32-3

Under hydrogen balloon room temperature, by 2-[4-[bis(2-methylpropyl)amino]-3-nitrophenoxy]-2-methylpropionic acid methyl ester (6g, 16.37mmol) and palladium carbon (0.9g) in methanol (20mL) stirring overnight.Through diatomaceous earth filtering mixture and filtrate vacuum concentration.With ethyl acetate/petroleum ether (1/20) as eluent, pass through silica gel column purification residue, obtain desired product (3.1g, 56% yield).

Step 4. Synthesis of 32-4

To a solution of 2-[3-amino-4-[bis(2-methylpropyl)amino]phenoxy]-2-methylpropanoic acid methyl ester (500mg, 1.49mmol) in tetrahydrofuran (10mL) was added triethylamine (451mg, 4.46mmol) and then 2,4-difluoro-1-phenylisocyanate (346mg, 2.23mmol). The resulting mixture was then stirred at room temperature for 3 hours. Water was added to quench the reaction and extracted with ethyl acetate. The organic layer was washed with water and brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/25) to obtain the desired product (570mg, 78% yield).

Step 5. Synthesis of 32

At room temperature, a mixture of 32-4 (650 mg, 1.32 mmol), lithium hydroxide (64 mg, 2.67 mmol) in tetrahydrofuran (8 ml) and water (4 mL) was stirred for 20 hours. The pH value of the solution was adjusted to 7 with aqueous hydrogen chloride solution (2N). The mixture was then extracted with ethyl acetate. The organic phase was washed with brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by preparative HPLC (column: X Bridge Shield RP18 OBD column, 5 μm, 19×150 mm; mobile phase A: water containing 0.05% ammonium bicarbonate, mobile phase B: acetonitrile; flow rate: 25 mL/min; gradient: 25% B-85% B in 8 minutes, detector: UV 254 nm). The collected fractions were concentrated to give the desired product (95.3 mg, 14% yield) as a white solid. LRMS(ES,m/z):478.3[M+H] ⁺ . ¹ HNMR(300MHz,DMSO-D ₆ ,ppm)δ9.29(s,1H),8.11(s,1H),7.92-7.84(m,1H),7.44(d,J＝3Hz,1H),7.34-7.26(m,1H),7.08–7.02(m,2H), 6.48(dd,J＝8.4Hz,J＝2.4Hz,1H),2.58(d,J＝6.9Hz,4H),1.63-1.58(m,2H),1.42(s,6H),0.83(d,J＝6.6Hz,12H).

Example 33

Step 1. Synthesis of 33-1

To the dichloromethane (12mL) solution of pyrimidine-5-amine (565mg, 5.94mmol) and diisopropylethylamine (1.046g, 8.09mmol) was added the dichloromethane (6mL) solution of carbonic acid bis(trichloromethyl) ester (601mg, 2.03mmol). The obtained mixture was stirred at room temperature for 15 minutes, and 32-3 (500mg, 1.49mmol) and triethylamine (902mg, 8.91mmol, 6.00 equivalents) were added successively. The reaction mixture was stirred at room temperature for 5 hours, then quenched with water by adding methanol. The reactant was extracted with dichloromethane, washed with water and brine, and dried over anhydrous sodium sulfate. After concentration, ethyl acetate/petroleum ether (1/5) was used as eluent by silica gel column purification residue to obtain the desired product (570mg, 84% yield).

Step 2. Synthesis of 33

A mixture of 33-1 (500 mg, 1.09 mmol), lithium hydroxide (53 mg, 2.21 mmol) in tetrahydrofuran (6 mL) and water (4 mL) was stirred overnight at room temperature. The pH of the solution was adjusted to 7 with aqueous hydrogen chloride (2N). The product was extracted with ethyl acetate, washed with brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by preparative HPLC (column: X Bridge Shield RP18 OBD column, 5 μm, 19×150 mm; mobile phase A: water containing 0.05% NH ₄ HCO ₃ , mobile phase B: ACN; flow rate: 25 mL/min; gradient: 25% B to 85% B in 8 minutes, detector: UV 254 nm). The collected fractions were concentrated to give the desired product (122.9 mg, 25% yield) as a white solid. LRMS(ES,m/z):444.4[M+H] ⁺ . ¹ HNMR(300MHz,DMSO-D ₆ ,ppm)δ9.92(s,1H),8.91(s,2H),8.82(s,1H),8.20(s,1H),7.58(d,J＝2.4Hz,1H),7.12(d,J＝8.4Hz, 1H), 6.49 (d, J = 8.4Hz, 1H), 2.62 (d, J = 6.6Hz, 4H), 1.62 (m, 2H), 1.47 (s, 6H), 0.85 (d, J = 6.6Hz, 12H).

Example 34

Step 1. Synthesis of 34-1

To a dichloromethane solution (12 mL) of 3-methyl-1,2-oxazole-5-amine (583 mg, 5.94 mmol) and diisopropylethylamine (1.04 g, 8.08 mmol) was added a dichloromethane (6 mL) solution of dichloromethyl carbonate (601 mg, 2.03 mmol). The resulting mixture was stirred at room temperature for 20 minutes. A dichloromethane (2 mL) solution of 32-3 (500 mg, 1.49 mmol) and triethylamine (902 mg, 8.91 mmol) were added, and the reaction mixture was stirred at room temperature for another 5 hours. Water and dichloromethane were added. The organic phase was separated, washed with brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column purification using ethyl acetate/petroleum ether (1:20) as eluent to obtain the desired product (580 mg, 85% yield).

Step 2. Synthesis of 34

At room temperature, 34-1 (500 mg, 1.09 mmol), lithium hydroxide (52 mg, 2.17 mmol) were stirred in a mixture of tetrahydrofuran (6 mL) and water (4 mL) overnight. The pH value of the solution was adjusted to 7 with aqueous hydrogen chloride solution (2N). The product was extracted with ethyl acetate, washed with brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by preparative HPLC (column: XBridge Shield RP18 OBD column, 5 μm, 19×150 mm; mobile phase A: water containing 0.05% ammonium bicarbonate, mobile phase B: acetonitrile; flow rate: 25 mL/min; gradient: 25% B-85% B in 8 minutes, detector: UV 254 nm). The collected fractions were concentrated to give the desired product (58.8 mg, 12% yield) as a white solid. LRMS(ES,m/z):447.3[M+H] ⁺ . ¹ HNMR:(300MHz,DMSO-D ₆ ,ppm)δ11.23(s,br,1H),8.25(s,1H),7.53(d,J＝2.4Hz,1H),7.10(d,J＝8.7Hz,1H),6.50(dd,J＝8.7Hz,J＝2.4Hz ,1H),5.95(s,1H),2.58(d,J＝6.6Hz,4H),2.17(s,3H),1.62-1.51(m,2H),1.46(s,6H),0.84(d,J＝6.3Hz,12H).

Example 35

Step 1. Synthesis of 35-1

At 110 ℃, 2-(4-fluoro-3-nitrophenoxy)-2-methylpropionic acid methyl ester (500mg, 1.95mmol), N-isobutylcyclohexylamine (450mg, 2.93mmol) and N, N-diisopropylethylamine (750mg, 5.85mmol) in dimethyl sulfoxide (10mL) solution was stirred overnight. The reactant was cooled to room temperature and diluted with ethyl acetate (100mL). The mixture was washed with water (60mL) and salt solution (60mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. Ethyl acetate/petroleum ether (1/8) was used as eluent to pass through a silica gel column purification residue to obtain the desired product (250mg, 33% yield).

Compound 35 was synthesized according to similar steps 3-5 in Example 32.

Example 35: LCMS (ES, m/z): 504.4 [M+H] ⁺ ; ¹ H NMR (300 MHz, DMSO-d ₆ , ppm): δ 9.38 (s, 1H), 8.27 (s, 1H), 7.91-7.83 (m, 1H), 7.55 (s, 1H), 7.34-7.25 (m, 1H), 7.08-6.98 (m, 2H), 6.44 (d, J=9.0 Hz, 1H), 2.83-2.62 (m, 3H), 1.96-1.81 (m, 2H), 1.78-1.61 (m, 2H), 1.58-1.26 (m, 8H), 1.23-1.11 (m, 5H), 0.82 (d, J=6.6 Hz, 6H).

Example 36

Step 1. Synthesis of 36-1

A 50 mL three-necked round-bottom flask was purged and maintained with an inert atmosphere of nitrogen and chlorosulfonic acid (19.6 g, 168.21 mmol) was placed therein. 1-Fluoro-2-nitrobenzene (10 g, 70.87 mmol) was then added dropwise at 65 ° C with stirring over 5 minutes. The resulting solution was stirred at 90 ° C overnight. The reaction was cooled to room temperature and then poured into 50 mL of water/ice. The mixture was extracted with 3 × 50 mL of dichloromethane. The organic layer was washed with 100 mL of saturated sodium bicarbonate, then with 2 × 100 mL of brine, dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain the desired product (9 g, 53% yield).

Step 2. Synthesis of 36-2

A 250 mL three-necked round-bottom flask was purged and maintained with an inert atmosphere of nitrogen and a solution of 4-fluoro-3-nitrobenzene-1-sulfonyl chloride (9 g, 37.56 mmol) in toluene (90 mL) was placed therein. PPh ₃ (29.5 g, 112.47 mmol) was then added in several batches over 60 minutes (exothermic). The resulting solution was stirred for 1 hour. Water (50 mL) was carefully added thereto while maintaining the reaction temperature below 45° C. The resulting solution was stirred at room temperature for 1 hour. The reaction mixture was extracted with 3×100 mL of dichloromethane. The combined organic layers were washed with 3×200 mL of brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:10-1:3) to give the desired product (4 g, 61% yield).

Step 3. Synthesis of 36-3

A 100 mL three-necked round-bottom flask was purged and maintained with an inert atmosphere of nitrogen. A solution of 4-fluoro-3-nitrobenzene-1-thiol (4 g, 23.10 mmol) and methyl 2-bromoacetate (4.24 g, 27.72 mmol) in N,N-dimethylformamide (50 mL) was placed. DIEA (5.97 g, 46.19 mmol) was then added dropwise over 10 minutes with stirring at 0°C. The resulting solution was stirred at 50°C overnight. The reaction mixture was cooled to room temperature and diluted with 50 mL _{of H₂O} . The reaction mixture was extracted with 3 x 50 mL of ethyl acetate. The combined organic layers were washed with 3 x 50 mL of brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:10-1:3) to obtain the desired product (4 g, 71% yield).

Step 4. Synthesis of 36-4

A 100 mL three-necked round-bottom flask, purged and maintained with an inert atmosphere of nitrogen, was charged with a solution of methyl 2-[(4-fluoro-3-nitrophenyl)thio]acetate (4 g, 16.31 mmol), bis(2-methylpropyl)amine (3.16 g, 24.45 mmol, 1.5 equivalents), and DIEA (4.2 g, 32.50 mmol, 2.0 equivalents) in DMSO (40 mL). The reaction was stirred at 80°C overnight. The mixture was cooled to room temperature. The resulting solution was diluted with 40 mL of _H₂O and extracted with 3 × 50 mL of ethyl acetate. The combined organic layers were washed with 3 × 50 mL of brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:10-1:3) to give the desired product (3.1 g, 54% yield).

Step 5. Synthesis of 36-5

In a 100mL round-bottom flask was placed a solution of 2-([4-[bis(2-methylpropyl)amino]-3-nitrophenyl]thio)methyl acetate (3 g, 8.46 mmol) and palladium on carbon (100 mg) in ethyl acetate (30 mL) and MeOH (5 mL). The flask was evacuated and flushed with nitrogen three times, then with hydrogen. The mixture was stirred at room temperature under a hydrogen atmosphere (balloon) for 3 hours. The solid was filtered off. The filtrate was concentrated in vacuo to give the desired product (2 g, 73% yield).

Step 6. Synthesis of 36-6

In a 100 mL three-necked round-bottom flask was placed a solution of methyl 2-([3-amino-4-bis(2-methylpropyl)amino]phenyl]thio)acetate (2 g, 6.16 mmol), 2,4-difluoro-1-isocyanatophenyl ester (1.15 g, 7.41 mmol) and triethylamine (1.25 g, 12.35 mmol) in tetrahydrofuran (50 mL). The reaction was stirred at room temperature overnight. The resulting mixture was concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:10-1:1) to give the desired product (1.5 g, 51% yield).

Step 7. Synthesis of 36

To a solution of methyl 2-([4-[bis(2-methylpropyl)amino]-3-[[(2,4-difluorophenyl)carbamoyl]amino]phenyl]thio)acetate (250 mg, 0.52 mmol) in ethanol (3 mL) and _H₂O (0.5 mL) was added sodium hydroxide (15% in water) (0.5 mL). The resulting solution was stirred at room temperature for 3 hours. The pH of the solution was adjusted to 6 with hydrogen chloride (1 N). The resulting mixture was concentrated in vacuo. The crude product was purified by preparative HPLC using the following conditions: column, Waters X-bridge RP 18, 19×150 mm, 5 μm; mobile phase, ACN/water (0.05% _NH₃H₂O ) 15%-40% over 6.5 minutes, flow rate: 20 mL/min; detector _, 254 nm. The desired product (60 mg, 25% yield) was obtained as a white solid. LCMS(ES,m/z):466[M+H] ⁺ .HNMR(300MHz,DMSO-d ₆ ,ppm): δ9.33(s,1H),8.10(s,1H),7.97-7.88(m,2H),7.30(t,J＝6.6Hz,1H),7.28(d,J＝6.0Hz,1H),7.17-7.02(m ,1H),6.94(dd,J＝8.4,2.4Hz,1H),3.38(s,2H),2.67(d,J＝6.9Hz,4H),1.69-1.60(m,2H),0.83(d,J＝6.6Hz,12H).

Example 37

Step 1. Synthesis of 37-1

To a solution of 4-bromo-1-fluoro-2-nitrobenzene (5 g, 22.73 mmol) in dimethyl sulfoxide (50 mL) was added bis(2-methylpropyl)amine (3.53 g, 27.31 mmol) and then N,N-diisopropylethylamine (3.53 g, 27.36 mmol). The reaction was then stirred at 100 ° C for 12 hours. After cooling to room temperature, the mixture was diluted with water (200 mL) and extracted with ethyl acetate (200 mL × 3). The organic phase was washed with brine (200 mL) and concentrated in vacuo to give the desired product (7 g, 94% yield).

Step 2. Synthesis of 37-2

A solution of 4-bromo-N,N-bis(2-methylpropyl)-2-nitroaniline (7 g, 21.26 mmol), benzylmercaptan (3.125 g, 25.16 mmol), _Pd2 (dba) _{3CHCl3 (} 2.2 g, 2.13 mmol), XantPhos (1.23 g, 2.12 mmol) and triethylamine (4.31 g, 42.67 mmol) in dioxane (100 mL) was stirred at 100°C for 2 hours. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/15-1/10) as eluent to give the desired product (4 g, 51% yield).

Step 3. Synthesis of 37-3

To a solution of 4-(benzylthio)-N,N-bis(2-methylpropyl)-2-nitroaniline (1 g, 2.68 mmol) in ethanol (20 mL) and water (2 mL) was added iron (750 mg, 13.43 mmol) followed by ammonium chloride (710 mg, 13.40 mmol). The reaction was then stirred at 80 ° C for 1 hour. After cooling to room temperature, the mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL × 3). The organic phase was concentrated in vacuo and the residue was purified by silica gel column with ethyl acetate/petroleum ether (1/10-1/3) as eluent to obtain the desired product (340 mg, 37% yield).

Step 4. Synthesis of 37-4

To a solution of 4-(benzylthio)-1-N,1-N-bis(2-methylpropyl)benzene-1,2-diamine (200 mg, 0.58 mmol) and triethylamine (71 mg, 0.70 mmol) in tetrahydrofuran (10 mL) was added 2,4-difluoro-1-phenylisocyanate (109 mg, 0.70 mmol). The reaction was then stirred at room temperature for 30 minutes. Water (10 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (15 mL × 3). The combined organic layers were washed with 3 × 50 mL of brine, dried over anhydrous sodium sulfate, and concentrated in vacuo to give the desired product (150 mg, 52% yield).

Step 5. Synthesis of 37-5

To a solution of 3-[5-(benzylthio)-2-[bis(2-methylpropyl)amino]phenyl]-1-(2,4-difluorophenyl)urea (150 mg, 0.30 mmol) in toluene (5 mL) was added aluminum chloride (398 mg, 2.98 mmol) in portions. The resulting mixture was then stirred at room temperature for 2 hours. The reaction was diluted with water (10 mL) and extracted with ethyl acetate (15 mL × 3). The combined organic layers were washed with 3 × 50 mL of brine, dried over anhydrous sodium sulfate, and concentrated in vacuo to give the desired product (70 mg, 57% yield).

Step 6. Synthesis of 37-6

To a solution of 3-[2-[bis(2-methylpropyl)amino]-5-thiophenyl]-1-(2,4-difluorophenyl)urea (70 mg, 0.17 mmol) and N,N-diisopropylethylamine (33.5 mg, 0.26 mmol) in dimethyl sulfoxide (1 mL) was added ethyl 2-bromo-2-methylpropanoate (36.7 mg, 0.19 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was diluted with water (5 mL) and extracted with ethyl acetate (5 mL × 3). The organic phase was washed with brine (5 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/5-1/3) as eluent to obtain the desired product (60 mg, 67% yield).

Step 7. Synthesis of 37

To a solution of ethyl 2-([4-[bis(2-methylpropyl)amino]-3-[[(2,4-difluorophenyl)carbamoyl]amino]phenyl]thio)-2-methylpropanoate (60 mg, 0.12 mmol) in tetrahydrofuran (1 mL) and water (0.2 mL) was added lithium hydroxide monohydrate (6.9 mg, 0.29 mmol). The reaction was stirred at 60°C for 12 hours. After cooling to room temperature, the mixture was concentrated in vacuo and the residue was purified by Flash-Prep-HPLC [column: C18 Waters X-bridge, 5 μm, 19×150 mm; mobile phase A: water (0.05% NH ₄ HCO ₃ ), mobile phase B: ACN; gradient: 50% ACN to 90% ACN over 10 minutes; detector: UV 254 nm] to give the desired product (12 mg, 21% yield) as a white solid. LCMS:(ES,m/z):494.2[M+H] ⁺ . ¹ HNMR:(300MHz,CD ₃ OD): δ8.03(s,1H),7.84-7.76(m,1H),7.20-7.12(m,2H),7.06-6.91(m,2H),2 .74(d,J＝14.1Hz,4H),1.78-1.69(m,2H),1.46(s,6H),0.87(d,J＝6.6Hz,12H).

Example 38

Step 1. Synthesis of 38-1

To a solution of 4-(benzylthio)-N1,N1-diisobutylbenzene-1,2-diamine (600 mg, 1.75 mmol) in tetrahydrofuran (10 mL) was added 5-isocyanatopyrimidine (862 mg, 7.12 mmol). Triethylamine (930 mg, 9.19 mmol) was then added. The reaction was stirred at room temperature for 2 hours. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL × 3). The organic phase was washed with brine (50 mL × 2), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (2/3) as eluent to obtain the desired product (600 mg, 74% yield).

Compound 38 was synthesized according to similar steps 5-7 in Example 37.

Example 38: LCMS (ES, m/z): 460.2 [M+H] ⁺ . 1H NMR (300 MHz, CD ₃ OD): δ 8.97 (s, 2H), 8.79 (s, 1H), 8.15 (s, 1H), 7.20 (s, 2H), 2.75 (d, J=6.9 Hz, 4H), 1.79-1.70 (m, 2H), 1.45 (s, 6H), 0.91 (d, J=6.6 Hz, 12H).

Example 39

Step 1. Synthesis of 39-1

To a solution of 4-(benzylthio)-N1,N1-diisobutylbenzene-1,2-diamine (175 mg, 0.51 mmol) in tetrahydrofuran (10 mL) was added 5-isocyanato-3-methylisoxazole (253 mg, 2.04 mmol) and then triethylamine (310 mg, 3.07 mmol). The reaction was then stirred at room temperature overnight, stirred at 45 ° C for 12 hours, and then stirred at 75 ° C for 36 hours. The mixture was cooled to room temperature and quenched by adding water/ice (50 mL). The mixture was extracted with ethyl acetate (20 mL × 3) and washed with brine (20 mL × 2). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give the desired product (100 mg, 42% yield).

Compound 39 was synthesized according to similar steps 5-7 in Example 37.

Example 39: LCMS (ES, m/z): 463.6 [M+H] ⁺ . ¹ H NMR: (300 MHz, CH ₃ OD, ppm): δ 8.15 (s, 1H), 7.20 (s, 2H), 6.06 (s, 1H), 2.73 (d, J=6.9 Hz, 4H), 2.24 (s, 3H), 1.77-1.66 (m, 2H), 1.45 (s, 6H), 0.89 (d, J=6.6 Hz, 12H).

Example 40

Step 1. Synthesis of 40-1

Cyclohexylamine (2 g, 20.17 mmol), 2,2-dimethyloxirane (1.45 g, 20.11 mmol) and ethanol (10 mL) were added to a 50 mL sealed tube. The resulting solution was stirred at 100 ° C for 2 days. After cooling to room temperature, petroleum ether (20 mL) was added and the solid was filtered out. The filtrate was concentrated in vacuo and the crude product was purified by silica gel column using methanol and dichloromethane (1:10) as eluent to obtain the desired product (2.4 g, 69% yield).

Step 2. Synthesis of 40-2

To a solution of 4-bromo-1-fluoro-2-nitrobenzene (1.45 g, 6.57 mmol) and diisopropylethylamine (1.70 g, 13.15 mmol) in dimethyl sulfoxide (10 mL) was added 40-1 (1.35 g, 7.88 mmol). The reaction mixture was stirred at 120 ° C for 1 day. Water (100 mL) was added to quench the reaction and the mixture was extracted with ethyl acetate (50 mL × 3). The combined organic layer was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column using ethyl acetate/petroleum ether (1/10) as eluent to obtain the desired product (1.42 g, 58% yield).

Compound 40 was synthesized according to similar steps 2-7 in Example 37.

Example 40: LRMS (ES, m/z) 536.4 [M+H] ^{+ .1} HNMR(300MHz,MeOD,ppm)8.14(s,1H),8.14-7.93(m,1H),7.27(d,J＝8.4Hz,1H),7.25-7.14(m,1H),7.07-6.92(m,2H),4.62(br,1H),3.14(br ,2H),2.61-2.60(m,1H),1.93(d,J＝10.8Hz,2H),1.74(d,J＝10.4Hz,2H ),1.57(d,J＝11.1Hz,1H),1.44(s,6H),1.36-1.03(m,5H),1.00(s,6H).

Example 41

Step 1. Synthesis of 41-1

At 110 ° C, a solution of N-isobutylcyclohexylamine (1.0 g, 6.45 mmol), 4-bromo-1-fluoro-2-nitrobenzene (1.42 g, 6.45 mmol) and N, N-diisopropylethylamine (1.66 g, 12.9 mmol) in dimethyl sulfoxide (20 mL) was stirred overnight. The reactants were cooled to room temperature and diluted with ethyl acetate (100 mL). The organic phase was washed with water (60 mL) and brine (60 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/10) as eluent to obtain the desired product (1.34 g, 59% yield).

Compound 41 was synthesized according to similar steps 2-7 in Example 37.

Example 41: LCMS (ES, m/z): 520.4 [M+H] ⁺ ; ¹ HNMR (300 MHz, DMSO-d ₆ ,ppm): δ12.72(brs,1H),9.40(s,1H),8.09(s,1H),8.02(s,1H),8.01-7.88 (m,1H),7.36-7.28(m,1H),7.17-7.14(m,1H),7.06-7.04(m,2H),2.79(d,J ＝6.0Hz,2H),2.62-2.55(m,1H),1.86-1.83(m,2H),1.78-1.66(m,2H),1.58 -1.48(m,1H),1.42-1.23(m,8H),1.21-0.98(m,4H),0.82(d,J=6.6Hz,6H).

Example 42

Step 1. Synthesis of 42-1

To a solution of 4-bromo-1-fluoro-2-nitrobenzene (6.1 g, 27.73 mmol) in DMSO (2 mL) was added diisopropylethylamine (7.2 g, 55.81 mmol) followed by N-ethyltetrahydropyran-4-amine (2.4 g, 18.58 mmol). After stirring overnight at 140° C., the reaction mixture was cooled to room temperature and water (60 mL) was added. The reaction mixture was extracted with ethyl acetate (60 mL×3). The combined organic layers were washed with brine (60 mL×2), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/40) to give N-(4-bromo-2-nitrophenyl)-N-ethyltetrahydropyran-4-amine (5.5 g, 60% yield).

42 was synthesized according to similar steps 2-7 in Example 37.

Example 42: LC-MS (ES, m/z): [M+H] ⁺ 494.4. ¹ HNMR (300MHz, DMSO-d ₆ ,ppm): δ9.43(s,1H),8.78(s,1H),8.27(d,J＝2.1Hz,1H),8.06-7.98(m,1H),7.34-7.26(m,1H),7.20(d,J＝8.1Hz,1H),7.07-7.00(m ,2H),3.82(d,J＝1.2Hz,2H),3.27-3.18(m,2H),3.03-2.95(m,3H),1.69(d,J＝6.6Hz,2H),1.45-1.22(m,8H),0.79(t,J＝6.9Hz,3H).

Example 43

Step 1. Synthesis of 43-1

To a solution of tetrahydropyran-4-one (7.5 g, 75 mmol) in tetrahydrofuran (60 mL) was added 2-methylpropane-1-amine (4.96 g, 68 mmol) at room temperature, followed by acetic acid (1.5 mL). The resulting solution was stirred at room temperature for 1 hour. NaBH(OAc) ₃ (28.83 g) was added thereto. After stirring at room temperature overnight, the reaction was quenched by the addition of water (50 mL), and the mixture was extracted with ethyl acetate (60 mL×3). The combined organic layers were washed with brine (60×3), dried over anhydrous sodium sulfate, and concentrated in vacuo to give N-(2-methylpropyl)tetrahydropyran-4-amine (6.2 g, crude product).

Compound 43 was synthesized according to similar steps 2-7 in Example 37.

Example 43: LC-MS (ES, m/z): 522.4 [M+H] ⁺ . ¹ H NMR (300 MHz, CDCl ₃ -d ₆ , ppm) δ 8.53 (s, 1H), 8.03 (s, 1H), 7.64-7.47 (m, 1H), 7.13-7.05 (m, 1H), 6.91 (t, 2H), 3.91 (d, J=11.1 Hz, 2H), 3.25-3.17 (m, 2H), 2.72-2.63 (m, 2H), 1.70-1.20 (m, 12H), 0.70 (d, J=6.3 Hz, 6H).

Example 44

Step 1. Synthesis of 44-1

To a solution of 2-methylpropane-1-amine (5.76 g, 78.76 mmol) and 1-methylpiperidin-4-one (9.80 g, 86.60 mmol) in dichloromethane (130 mL) was added sodium cyanoborohydride (50.1 g, 236.39 mmol) in portions. The reaction was then stirred at room temperature for 16 hours. The solid was filtered off and the mixture was diluted with water (500 mL). The pH value of the mixture was adjusted to 9 with sodium bicarbonate. The mixture was extracted with dichloromethane (500 mL × 4). The organic phase was washed with brine (1000 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain the desired product (6 g, 45% yield).

Step 2. Synthesis of 44-2

At 100 ° C, a solution of 1-methyl-N-(2-methylpropyl)piperidin-4-amine (1.98 g, 11.64 mmol), 4-bromo-1-fluoro-2-nitrobenzene (1.7 g, 7.73 mmol) and N,N-diisopropylethylamine (3.01 g, 23.30 mmol) in dimethyl sulfoxide (20 mL) was stirred for 17 hours. After cooling to room temperature, the mixture was diluted with water (200 mL) and extracted with ethyl acetate (200 mL × 3). The organic phase was washed with brine (500 mL × 2), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by Flash-Prep-HPLC under the following conditions: [column: silica gel; mobile phase A: petroleum ether, mobile phase B: ethyl acetate; gradient: 0% ethyl acetate-100% ethyl acetate in 25 minutes; detector: UV 254 nm] to obtain the desired product (1.2 g, 42% yield).

Compound 44 was synthesized according to similar steps 2-7 in Example 37.

Example 44: LCMS (ES, m/z): 535.4 [M+H] ⁺ ; ¹ H NMR (300 MHz, DMSO-d ₆ ,ppm): δ9.45(s,1H),8.29(s,1H),8.12(d,J＝2.1Hz,1H),7.98-7.90(m,1H),7.35-7.28(m,1H),7.20(d,J＝8.1Hz,1H),7.09-7.02(m,2 H),2.80-2.72(m,4H),2.66-2.58(m,1H),2.11(s,3H),1.85-1.73(m,4H),1.57-1.46(m,2H),1.36-1.29(m,7H),0.82(d,J＝6.6Hz,6H).

Example 45

Step 1. Synthesis of 45-1

To a dichloromethane (75mL) solution of cyclohexanone (7.8g, 79.48mmol) and 2-methoxyethane-1-amine (5g, 66.57mmol) was added acetic acid (0.1mL). The reaction was stirred at 25°C for 0.5 hour. Sodium cyanoborohydride (16.7g, 265.75mmol) was added and the mixture was stirred for another 5 hours at 25°C. Saturated ammonium chloride solution (50mL) was added to quench the reaction. The resulting mixture was extracted with dichloromethane (50mL×2) and washed with brine (50mL×2). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. Ethyl acetate/petroleum ether (1/4-100/1) was used as eluent to pass through a silica gel column purification residue to obtain the desired product (6g, 57% yield).

Step 2. Synthesis of 45-2

To a solution of N-(2-methoxyethyl)cyclohexylamine (5 g, 31.80 mmol) and 4-bromo-1-fluoro-2-nitrobenzene (8.4 g, 38.18 mmol) in dimethyl sulfoxide (30 mL) was added N,N-diisopropylethylamine (6.2 g, 47.97 mmol). The reaction was then stirred at 100°C for 16 hours. The reaction was cooled to room temperature, diluted with water (200 mL), and extracted with ethyl acetate (200 mL x 2). The organic phase was washed with water (100 mL x 2) and brine (100 mL x 2), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by Flash-Prep-HPLC under the following conditions: [column: C18 silica gel; mobile phase A: water (0.05% TFA), mobile phase B: ACN, gradient: 55% ACN-100% ACN; detector: UV 254 nm] to give the desired product (8 g, 70% yield).

Compound 45 was synthesized according to similar steps 2-7 as in Example 37.

Example 45: LCMS (ES, m/z): 522.2 [M+H] ⁺ . ¹ H NMR: (300 MHz, DMSO-d ₆ , ppm): δ 9.40 (s, 1H), 8.62 (s, 1H), 8.19 (d, J=1.8 Hz, 1H), 8.01-7.93 (m, 1H), 7.35-7.19 (m, 2H), 7.08-7.01 (m, 2H), 3.25-3.14 (m, 4H), 3.12 (s, 3H), 2.62-2.80 (m, 1H), 1.80-1.98 (m, 2H), 1.79-1.60 (m, 2H), 1.59-1.43 (m, 1H), 1.32 (s, 6H), 1.20-0.90 (m, 5H).

Example 46

Step 1. Synthesis of 46-1

At 110 ° C, a solution of N-(2-methylpropyl)tetrahydrothiopyran-4-amine (3.1 g, 17.89 mmol), 4-bromo-1-fluoro-2-nitrobenzene (3.94 g, 17.91 mmol) and N, N-diisopropylethylamine (4.62 g, 35.75 mmol) in dimethyl sulfoxide (30 mL) was stirred overnight. After cooling to room temperature, the reaction was diluted with ethyl acetate (200 mL). The organic phase was washed with water (80 mL) and brine (80 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/10) as eluent to obtain the desired product (1.5 g, 22% yield).

Step 2. Synthesis of 46-2

To N-(4-bromo-2-nitrophenyl)-N-(2-methylpropyl) tetrahydrothiopyran-4-amine (1.5g, 4.02mmol) in dichloromethane (20mL) solution, add 3-chloroperbenzoic acid (2.1g, 12.17mmol).The reaction was stirred at room temperature for 2 hours, and then the mixture was diluted with dichloromethane (50mL).The reactant was washed with saturated sodium bisulfite solution (30mL), water (30mL) and salt solution (30mL).The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo.With ethyl acetate/petroleum ether (1/5) as eluent, the desired product (0.6g, 37% yield) was obtained by silica gel column purification residue.

Step 3. Synthesis of 46-3

A solution of 46-2 (200 mg, 0.49 mmol), phenylmethylthiol (68 mg, 0.55 mmol), Pd ₂ (dba) ₃ .CHCl ₃ (26 mg, 0.03 mmol), XantPhos (30 mg, 0.05 mmol), and triethylamine (76 mg) in 1,4-dioxane (3 mL) was stirred at 100° C. for 1 hour. The mixture was then diluted with ethyl acetate (30 mL), and the solid was filtered off. The filtrate was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column using ethyl acetate/petroleum ether (1/5) as eluent to give the desired product (150 mg, 68% yield).

Step 4. Synthesis of 46-4

To 46-3 (150mg, 0.33mmol) acetic acid (5mL) solution, add iron (187mg, 3.35mmol), then at room temperature stirring reaction 0.5 hour. Dilute the mixture with ethyl acetate (50mL). Filter out solid, filtrate is washed with saturated sodium carbonate solution (30mL) and salt solution (30mL). Organic phase is dried over anhydrous sodium sulfate and concentrated in vacuo. Use ethyl acetate/petroleum ether (1/5) as eluent by silica gel column purification residue, obtain desired product (130mg, 93% yield).

Step 5. Synthesis of 46-5

At room temperature, a tetrahydrofuran (3 mL) solution of 46-4 (130 mg, 0.31 mmol), 2,4-difluoro-1-phenyl isocyanate (73 mg, 0.47 mmol) and triethylamine (95 mg, 0.94 mmol) was stirred for 30 minutes. The mixture was diluted with ethyl acetate (30 mL) and washed with water (20 mL) and salt solution (20 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. Ethyl acetate/petroleum ether (1/2) was used as eluent to obtain the desired product (120 mg, 67% yield).

Step 6. Synthesis of 46-6

To a solution of 46-5 (120 mg, 0.21 mmol) in toluene (5 mL) was added _AlCl₃ (240 mg, 1.80 mmol) and the reaction was stirred at room temperature for 30 minutes. The mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL×2). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give the desired product (80 mg, 79% yield).

Step 7. Synthesis of 46-7

At room temperature, 2-bromo-2-methylpropionic acid ethyl ester (51mg, 0.26mmol), 46-6 (80mg, 0.17mmol) and N, N-diisopropylethylamine (44mg, 0.34mmol) in dimethyl sulfoxide (3mL) solution was stirred overnight. The reactant was diluted with ethyl acetate (20mL) and washed with water (10mL) and salt solution (10mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. Ethyl acetate/petroleum ether (1/5) was used as eluent by silica gel column purification residue to obtain the desired product (70mg, 71% yield).

Step 8. Synthesis of 46

To a solution of 46-7 (70 mg, 0.12 mmol) in ethanol (1.5 mL) and water (0.5 mL) was added LiOH (160 mg, 6.68 mmol). The resulting mixture was stirred at 75°C for 1 hour. The reaction was then cooled to room temperature and diluted with water (20 mL). The mixture was extracted with ethyl acetate (30 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by preparative HPLC using the following conditions: [column: X Bridge Shield RP18 OBD, 5 μm, 19×150 mm; mobile phase A: water (10 mmol/L NH ₄ HCO ₃ ), mobile phase B: ACN; gradient: 15% ACN to 40% over 8 minutes; detector: UV 254 nm] to give the desired product (27.6 mg, 41% yield). LCMS: (ES, m/z):570.2[M+H] ⁺ ; ¹ H NMR (300MHz, DMSO-d ₆ ):ppmδ9.48(s,1H),8.18(s,2H),8.00-7.92(m,1H),7.36-7.28(m,1H),7.21-7.18(m,1H),7.09-7.03(m,2H),3.24-3.15(m ,2H),3.10-2.93(m,3H),2.90-2.73(m,2H),2.26-2.21(m,2H),2.01-1.89(m,2H),1.39-1.33(m,7H),0.82(d,J＝6.6Hz,6H).

Example 47

Step 1. Synthesis of 47-1

At room temperature, potassium carbonate (552 mg, 4 mmol) was added to a solution of 9-3 (346 mg, 1 mmol) in chloroform (12 mL). After the reaction was cooled to 0 ° C, a solution of thiophosgene (230 mg, 2 mmol) in chloroform (8 mL) was added dropwise. The reaction mixture was stirred at 0 ° C for 3 hours. The solid was filtered and the filtrate was concentrated. The crude product was purified by preparative TLC plate (petroleum ether/ethyl acetate=10/1) to obtain the desired product (180 mg, 46% yield).

Step 2. Synthesis of 47-2

A solution of 47-1 (194 mg, 0.5 mmol) in acetonitrile (10 mL) and 1,2-diaminobenzene (54 mg, 0.5 mmol) was stirred at room temperature for 16 hours. N,N-diisopropylethylamine (129 mg, 1 mmol) and HATU (285 mg, 0.75 mmol) were added and the reaction was stirred at room temperature for another hour. The reaction was diluted with ethyl acetate (50 mL). The organic layer was washed with water (50 mL) and brine (50 mL), dried over sodium sulfate, filtered and concentrated. The crude product was purified by reverse phase HPLC (acetonitrile/0.05% TFA. = 10%-95%) to give the desired product (110 mg, 47% yield).

Step 3. Synthesis of 47

At 60 ℃, by 47-2 (110mg, 0.24mmol) and lithium hydroxide (0.7mL, 0.7mmol, the 1M aqueous solution) solution in tetrahydrofuran/methanol (1.4mL, v/v=1/1) stirred 24 hours.Reactant is cooled and with 1N HCl the pH value of solution is adjusted to 6.Dilute reactant with ethyl acetate (50mL).Organic layer is washed with water (50mL) and salt solution (50mL), dried over sodium sulfate, filtered and concentrated.By reversed-phase HPLC (acetonitrile/0.02% ammonium hydroxide aqueous solution=10%-95%) purifying residue, obtain desired product (11.73mg, 11% productive rate). LCMS(ES,m/z):449.2[M+H] ⁺ .HNMR(400MHz,DMSO-d ₆ ,ppm): δ12.15-12.08(m,1H),11.73(s,1H),8.64(s,1H),8.23(s,1H),7.30-7.16(m,3H),6.97-6.87(m 3H),2.59-2.48(m,6H),1.78-1.56(m,8H),0.86(d,J＝6.4Hz,12H).

Example 48

Step 1. Synthesis of 48-1

A solution of 9-3 (121 mg, 0.35 mmol), iodobenzene (143 mg, 0.7 mmol), Pd ₂ (dba) ₃ (16 mg, 0.018 mmol), PCy ₃ (10 mg, 0.035 mmol) and sodium tert-butoxide (50 mg, 0.5 mmol) in toluene (5 mL) was stirred at 120° C. under microwave conditions for 3 hours. Ethyl acetate (50 mL) was added and the reactants were washed with water (50 mL) and brine (50 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The residue was purified by pre-TLC (petroleum ether/ethyl acetate = 10/1) to give the desired product (53 mg, 36% yield).

Step 2. Synthesis of 48

At room temperature, lithium hydroxide (0.5mL, 0.5mmol, 1N) is added to a solution of 48-1 (53mg, 0.125mmol) in tetrahydrofuran/methanol (1mL, v/v=1/1). The reaction mixture is stirred at 60°C for 24 hours. The reaction is cooled and the pH of the solution is adjusted to 6 with 1NHCl. The reactant is diluted with ethyl acetate (50mL). The organic layer is washed with water and brine, dried over sodium sulfate, filtered and concentrated. By pre-TLC (petroleum ether/ethyl acetate=7/1) purification of residue, the desired product (40mg, 78% yield) is obtained. LCMS(ES,m/z):409.2[M+H] ⁺ .HNMR(400MHz,CD ₃ OD,ppm): δ7.35(d,J＝2.0Hz,1H),7.24-7.20(m,2H),7.10-7.05(m,3H),6.87-6.84(m 2H),2.58-2.53(m,6H),1.83-1.80(m,2H),1.71-1.67(m,6H),0.87(d,J＝6.4Hz,12H).

Example 49

Step 1. Synthesis of 49-1

A solution of 9-3 (277 mg, 0.8 mmol), 2-bromopyrimidine (254 mg, 1.6 mmol), Pd ₂ (dba) ₃ (37 mg, 0.04 mmol), PCy ₃ (22 mg, 0.08 mmol), and sodium tert-butoxide (115 mg, 1.2 mmol) in toluene (5 mL) was stirred at 150° C. under microwave conditions for 3 hours. Ethyl acetate (50 mL) was added, and the reactants were washed with water (50 mL) and brine (50 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated. The residue was purified by preparative TLC (petroleum ether/ethyl acetate = 10/1) to give the desired product.

Step 2. Synthesis of 49

At room temperature, lithium hydroxide (2.4mL, 2.4mmol, 1N) is added in tetrahydrofuran/methanol (4.8mL, v/v=1/1) solution of 49-1 (40mg, 0.8mmol).Reaction mixture is stirred at 60 ℃ for 24 hours.Reactant is cooled and the pH value of solution is adjusted to 6 with 1N HCl.The reactant is diluted with ethyl acetate (50mL).The organic layer is washed with water and salt water, dried over sodium sulfate, filtered and concentrated.By HPLC purification of residue, desired product (8.92mg, 19% yield) is obtained. LCMS(ES,m/z):411.2[M+H] ⁺ .HNMR(400MHz,CD ₃ OD,ppm): δ8.59(s,1H),8.44(d,J＝4.8Hz,2H),7.19(d,J＝8.4Hz,1H),7.01(d,J＝8.0Hz,1H),6.79( t,J＝4.8Hz,1H),2.65-2.58(m,6H),1.93-1.91(m,2H),1.74-1.65(m,6H),0.91(d,J＝6.4Hz,12H).

Example 50

Step 1. Synthesis of 50-1

At 0 ℃, sulfuric acid (2mL) is added to the methanol (4mL) solution of 25-2 (500mg, 1.4mmol).The reaction is stirred at 80 ℃ for 16 hours.The mixture is extracted with ethyl acetate (50mL) and washed with sodium bicarbonate solution (100mL).The organic layer is dried over sodium sulfate, filtered and concentrated.The residue is purified by silica gel column chromatography (petroleum ether/ethyl acetate=4/1) to obtain the desired product (130mg, 23% yield).

Step 2. Synthesis of 50-2

To a solution of 50-1 (80 mg, 0.2 mmol) in methanol (5 mL) was added palladium carbon (40 mg). The reaction was stirred under hydrogen at room temperature for 2 hours. The mixture was filtered and the filtrate was concentrated to give the desired product (50 mg, yield 69%).

Step 3. Synthesis of 50-3

To a solution of 50-2 (50 mg, 0.14 mmol) in dichloromethane (5 mL) was added triethylamine (43 mg, 0.42 mmol) and then isobutyryl chloride (30 mg, 0.28 mmol). The reaction was stirred at room temperature for 16 hours. The mixture was extracted with ethyl acetate (50 mL) and washed with water (50 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The residue was purified by pre-TLC (petroleum ether/ethyl acetate=4/1) to obtain the desired product (30 mg, 50% yield).

Step 4. Synthesis of 50

To a solution of 50-3 (30 mg, 0.07 mmol) in methanol (1 mL) and tetrahydrofuran (1 mL) was added lithium hydroxide solution (1 mL, 1 N). The reaction was stirred at 60 ° C for 16 hours. The mixture was acidified with hydrochloric acid (1 N) and then extracted with ethyl acetate (50 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The residue was purified by preparative HPLC to obtain the desired product as a white solid (10 mg, 34% yield). LCMS (ES, m / z): 417.2 [M + H] ⁺ . HNMR (400 MHz, DMSO-d ₆ ,ppm): δ8.83(s,1H),8.24(d,J＝2.0Hz,1H),7.24(d,J＝8.4Hz,1H),6.92(dd,J1＝8.4Hz,J2＝ 2.0Hz,1H),3.82-3.79(m,2H),3.17(t,J＝11.2Hz,2H),2.80-2.73(m,3H),2.68-2.61(m,2H ),2.59-2.54(m,1H),2.37-2.29(m,2H),1.88-1.86(m,1H),1.77-1.73(m,1H),1.62-1.59( m,2H),1.50-1.42(m,2H),1.28-1.23(m,1H),1.11(d,J＝6.8Hz,6H),0.79(d,J＝6.4Hz,6H).

Example 51

Step 1. Synthesis of 78-1

To a stirred solution of 1,2-dichloroethane (5 mL) of 50-2 (105 mg, 0.29 mmol) was added cyclopentanone (90 mg, 1.07 mmol) and trifluoroacetic acid (90 mg, 0.79 mmol). The mixture was stirred at room temperature for 1 hour, and then tetramethylammonium triacetoxyborohydride (150 mg, 0.57 mmol) was added. The mixture was stirred at 60 ° C for 16 hours. After cooling, the reaction mixture was extracted with ethyl acetate (50 mL). The organic phase was washed with NaHCO ₃ aqueous solution (20 mL) and brine (10 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by pre-TLC (eluent: petroleum ether: ethyl acetate = 4: 1) to obtain the desired product (50 mg, 40% yield).

Step 2. Synthesis of 78

To a solution of 78-1 (50 mg, 0.12 mmol) in tetrahydrofuran (1.5 mL) and methanol (1.5 mL) was added lithium hydroxide (1 M, 1.5 mL, 1.5 mmol). The mixture was stirred at 60 ° C for 3 hours. After cooling, the reactant was acidified to pH = 4 with hydrochloric acid (1 N) and extracted with EtOAc. The organic phase was washed with brine (10 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo to give the desired product (22 mg, 44% yield). LCMS(ES,m/z):415.3[M+H] ⁺ . ¹ HNMR:(400MHz,CD ₃ OD,ppm): δ7.00(d,J＝8.0Hz,1H),6.59-6.54(m,2H),3.91(d,J＝11.2Hz,2H),3.79-3.77(m,1H),3.32-3.26(m,1H),3.26-3.21(m ,1H),2.98(d,J＝9.6Hz,1H),2.78-2.71(m,3H),2.56-2.41(m,3H),2.00-1.92(m,3H),1.86-1.37(m,12H),0.82(dd,J1＝24.8Hz,

Example 52

Step 1. Synthesis of 83-1

At 0 ° C, sodium hydride (8.6 g, 216 mmol, 60% oil solution) was added to a solution of 2-chloro-5-pyridineacetonitrile (14.3 g, 94 mmol) in dimethylformamide (120 mL) in batches over 20 minutes. The mixture was stirred for another 20 minutes and 1,3-dibromopropane (20 g, 98.7 mmol) was added. The reaction mixture was heated to room temperature and stirred for 2 hours. The reaction was quenched with water (50 mL). Ethyl acetate (200 mL) was added and the organic phase was washed with water (100 mL) and brine (100 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether-petroleum ether/ethyl acetate=3/1) to obtain the desired product (9.4 g, 52% yield).

Step 2. Synthesis of 83-2

At room temperature, sodium methoxide (3.6 g, 20 mmol, 30% methanol solution) and 83-1 (960 mg, 5 mmol) were added to a solution of potassium hydroxide (840 mg, 15 mmol) in water/methanol (6 mL, v/v=1/2). The mixture was then heated to 100° C. and reacted for 48 hours. After cooling to 0° C., the mixture was adjusted to pH~5 with 1N HCl. The mixture was diluted with water (50 mL) and extracted with dichloromethane (50 mL×4). The organic layer was dried over sodium sulfate, filtered and concentrated to give the desired product (950 mg, 92% yield).

Step 3. Synthesis of 83-3

At 0 ° C, concentrated nitric acid (1 mL) was added dropwise to a solution of 83-2 (350 mg, 1.7 mmol) in concentrated sulfuric acid (2 mL). The reaction was heated to 50 ° C for 16 hours. After cooling to room temperature, the mixture was poured into ice water and the pH value of the mixture was adjusted to 4 with 50% sodium hydroxide at 0 ° C. The mixture was extracted with dichloromethane (50 mL × 3). The organic layer was dried over sodium sulfate, filtered and concentrated. The residue was purified by reverse phase HPLC (MeCN/0.05% TFA aqueous solution = 5% to 95%) to obtain the desired product (180 mg, 42% yield).

Step 4. Synthesis of 83-4

At 0 ° C, to a solution of 83-3 (504 mg, 2 mmol) in thionyl chloride (3 mL) was added dimethylformamide (292 mg, 4 mmol) dropwise. The reaction was heated to 80 ° C for 16 hours and then concentrated. The residue was dissolved in dichloromethane (5 mL) and then methanol (1 mL) was added at 0 ° C. The mixture was stirred at room temperature for 0.5 hours. The mixture was concentrated and purified by reverse phase HPLC (MeCN/0.05% TFA aqueous solution = 5% to 95%) to obtain the desired product (400 mg, 74% yield).

Step 5. Synthesis of 83-5

To a solution of 83-4 (135 mg, 0.5 mmol) in N-methyl-2-pyrrolidone (2 mL) was added N,N-diisopropylethylamine (97 mg, 0.75 mmol) and diisobutylamine (97 mg, 0.75 mmol). The reaction was stirred at 90° C. for 16 hours. The mixture was directly purified by reverse phase HPLC (MeCN/0.05% TFA aqueous solution = 5% to 95%) to give the desired product (168 mg, 93% yield).

Step 6. Synthesis of 83-6

To 83-5 (554mg, 1.5mmol) in methanol (10mL) solution, add 10% palladium carbon (110mg).Stirring reaction 2 hours under hydrogen room temperature.Filter mixture and concentrate filtrate.By silica gel column chromatography (petroleum ether-petroleum ether/ethyl acetate=5/1) purification of crude product, obtain desired product (400mg, 80% yield).

Step 7. Synthesis of 83-7

At 0 ° C, triethylamine (50 mg, 0.48 mmol) and 2,4-difluoro-1-phenyl isocyanate (75 mg, 0.48 mmol) were added to a solution of 83-6 (80 mg, 0.24 mmol) in tetrahydrofuran (10 mL). The reaction was stirred at room temperature for 2 hours. The mixture was extracted with ethyl acetate (50 mL) and washed with water (50 mL). The organic phase was concentrated and purified by Pre-TLC (petroleum ether/ethyl acetate = 6/1) to obtain the desired product (80 mg, 68% yield).

Step 8. Synthesis of 83

To a solution of 83-7 (80 mg, 0.16 mmol) in methanol (2 mL) and tetrahydrofuran (2 mL) was added lithium hydroxide solution (2 mL, 1 M, 2 mmol). The reaction was stirred at 60 ° C for 5 hours. The mixture was acidified with hydrochloric acid (1 N) and extracted with ethyl acetate (50 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The residue was purified by Pre-TLC (eluent: petroleum ether: ethyl acetate = 1: 1) to obtain the desired product (65.2 mg, white solid, 86% yield). LCMS(ES,m/z):475.3[M+H] ⁺ .HNMR (400MHz, DMSO-d6, ppm): δ12.41 (brs, 1H), 9.28 (s, 1H), 8.05-7.94 (m, 3H), 7.86 (d, J = 2.0Hz, 1H), 7.32-7.26 (m, 1H), 7.03-6.99 (m, 1H), 2.96(d,J＝6.8Hz,4H),2.67-2.60(m,2H),2.38-2.31(m,2H),1.95-1.8 6(m,1H),1.80-1.75(m,1H),1.73-1.66(m,2H),0.76(d,J＝6.4Hz,12H).

Example 53

Step 1. Synthesis of 120-1

To a solution of 83-4 (1.7 g, 6.3 mmol) in dimethyl sulfoxide (30 mL) was added N-ethyltetrahydropyran-4-amine (1.6 g, 12.6 mmol) and N,N-diisopropylethylamine (1.6 g, 12.6 mmol). The reaction was stirred at 90 ° C for 16 hours. After cooling, the reactant was diluted with ethyl acetate (100 mL) and washed with water (100 mL) and brine (100 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether-petroleum ether/ethyl acetate=4/1) to obtain the desired product (1.6 g, 69% yield) as a yellow oil.

Step 2. Synthesis of 120-2

To 120-1 (1.6g, 4.4mmol) in methanol (50mL) solution, add 10% palladium carbon (320mg).Stirring reaction 2 hours under hydrogen room temperature.Filter the mixture and concentrate.By silica gel column chromatography (petroleum ether-petroleum ether/ethyl acetate=1/1) purification of crude product, obtain desired product (950mg, 65% yield), be light grey oily thing.

Step 3. Synthesis of 120-3

To a mixture of 120-2 (80 mg, 0.24 mmol), 1-chloro-4-iodobenzene (114 mg, 0.48 mmol) and sodium tert-butoxide (46 mg, 0.48 mmol) in methylbenzene (4 mL) was added tris (dibenzylideneacetone) dipalladium (0) (11 mg, 0.012 mmol) and 2-dicyclohexylphosphino-2', 6'-diisopropoxybiphenyl (11 mg, 0.024 mmol). The mixture was stirred in microwaves at 120 ° C for 30 minutes under N _2. After the reaction was completed, water (10 mL) was added, and the reaction mixture was extracted with ethyl acetate (20 mL). The organic phase was washed with brine (10 mL), dried over sodium sulfate and concentrated. The crude product was purified by preparative TLC (eluent: petroleum ether/ethyl acetate=1/1) to obtain the desired product (45 mg, 42% yield) as a yellow gel.

Step 4. Synthesis of 120

To a mixture of 120-3 (45 mg, 0.1 mmol) in tetrahydrofuran (1.5 mL) and methanol (1.5 mL) was added lithium hydroxide (1 M, 1.5 mL), the mixture was stirred at 60 ° C for 3 h, and after the reaction was complete, the mixture was acidified to pH = 3 with hydrochloric acid (1 M) and extracted with ethyl acetate (20 mL). The organic phase was washed with brine (10 mL), dried over sodium sulfate and concentrated. The crude product was purified by preparative TLC (eluent: dichloromethane/methanol=10/1) to obtain the desired product (32 mg, 73% yield) as a white solid.

Example 54: Inhibition of Kynurenine Production in HeLa Cells

The compounds described herein were assayed for their inhibitory effects on kynurenine production in HeLa cells (derived from human cervical carcinoma). Exemplary results are shown in Table 2. HeLa cells were seeded at a density of 5 × 10 ³ cells/well in 96-well plates in RPMI1640/phenol red-free medium containing 2 mM L-glutamine and 10% fetal bovine serum (FBS, GIBCO) and grown overnight in a 5% CO ₂ , 37°C incubator. After 24 hours, human IFN-γ (GIBCO) (final concentration 50 ng/mL) and a test compound solution (serially diluted for each well) were added to each well, resulting in a final volume of 200 μL per well. After 48 hours of incubation with the compounds, 140 μL of supernatant was removed from each well and transferred to a new 96-well plate. 10 μL of 6.1N trichloroacetic acid was added to each well, mixed, and incubated at 50°C for 30 minutes. The reaction mixture was then centrifuged at 2500 rpm for 10 minutes, and 100 μL of the supernatant per well was transferred to another 96-well plate and mixed with 100 μL of a 2% (w/v) solution of p-dimethylaminobenzaldehyde in acetic acid. The yellow color derived from kynurenine was measured at 480 nm using a SPECTRAmax i3 reader. Each compound concentration was measured in triplicate, and IC50 values were calculated using nonlinear regression using Graphpad Prism 5.0.

The inhibitory activities of representative compounds described herein compared to another IDO inhibitor, such as INCB-24360, and controls 1 and 2 are shown in Table 2.

Table 2. Inhibitory activity of representative compounds

*: Control 1: 3-(3-(3-(2,4-difluorophenyl)ureido)-4-(diisobutylamino)phenyl)-4,4,4-trifluorobutanoic acid. See Example 30 of WO2014/150646

**: Control 2: (1R,2S)-2-(4-(diisobutylamino)-3-(3-(3-methylisoxazol-5-yl)ureido)phenyl)cyclopropanecarboxylic acid. See Example 30 of WO2014/150677

Example 55: Inhibition of Kynurenine Production in SKOV-3 Cells

The inhibitory effects of the compounds described herein on kynurenine production in SKOV-3 cells (derived from human ovarian carcinoma) were determined using similar experimental procedures as described in Example A. Exemplary results are shown in FIG1 .

The assay was performed in a similar manner as described in Example A, except that SKOV-3 cells were cultured in DMEM medium containing 10% FBS. FIG1 shows the _IC50 of INCB-24360 and Compound 9 measured at 10.2 nM and 7.4 nM, respectively. The structure of Compound 9 is shown below:

Example 56: Inhibition of LPS-induced plasma kynurenine in mice

The compounds described herein were tested for their inhibitory effects on lipopolysaccharide (LPS)-induced plasma kynurenine in mice. Exemplary results are shown in FIG2 . Female Balb/c mice (approximately 20 g, obtained from Vital River Laboratory Animals, Inc.) were treated intraperitoneally with either a vehicle control (normal saline) or 5 mg/kg LPS, followed by oral administration of 30 mpk of compound 9 within 5 minutes of LPS injection. Blood samples (500 μL) were collected via retro-orbital puncture into K ₂ EDTA tubes at 12 and 24 hours (terminal bleeding) after treatment with LPS or LPS plus compound 9. For the vehicle control group, plasma samples were collected 24 hours after administration. Blood samples were placed on ice and immediately centrifuged at 4° C. (2000 g, 5 minutes) to obtain plasma. Plasma kynurenine levels were determined by LC-MS/MS analysis. Each group contained 3 mice, and the average plasma kynurenine levels are plotted in FIG2 . Figure 2 shows LPS-induced plasma kynurenine levels in mice compared to baseline (vehicle control group), and that concomitant treatment with LPS and IDO inhibitor Compound 9 reduced plasma kynurenine to levels below baseline, with decreases of 72% and 85% at the 12 hour and 24 hour time points, respectively, compared to treatment with LPS alone.

Example 57: Human Hepatocyte Clearance Study

The in vitro human hepatocyte clearance of the compounds described herein was studied using harvested human hepatocytes purchased from Bioreclamation IVT (Westbury, NY, catalog number X008001, Lot # TQJ). The assay was performed according to the manufacturer's instructions. Briefly, 10 mM stocks of test compounds and positive control (verapamil) were prepared in 100% DMSO. Thaw medium (50 mL) used in the study contained: 31 mL Williams E medium (GIBCO catalog number 12551-032); 15 mL isotonic percoll (GE Healthcare catalog number 17-0891-09); 500 μL 100× GlutaMax (GIBCO catalog number 35050); 750 μL HEPES (GIBCO catalog number 15630-080); 2.5 mL FBS (Corning catalog number 35-076-CVR); 50 μL human insulin (GIBCO catalog number 12585-014) and 5 μL dexamethasone (NICPBP). The culture medium was made by supplementing Williams E medium with 1× GlutaMax. Thaw medium and culture medium (serum-free) were placed in a 37°C water bath for at least 15 minutes before use. Compound stocks were diluted to 100 μM by combining 198 μL of 50% acetonitrile/50% water and 2 μL of a 10 mM stock solution. Verapamil was used as a positive control in the assay. The vial containing the cryopreserved hepatocytes was removed and thawed in a 37°C water bath with gentle shaking. The contents of the vial were poured into a 50 mL conical tube of thawing medium. The vial was centrifuged at 100 g for 10 minutes at room temperature. The thawing medium was aspirated and the hepatocytes were resuspended in serum-free medium to produce approximately 1.5 × 10 ⁶ cells/mL. Hepatocyte viability and density were counted using trypan blue exclusion, and the cells were then diluted to a working cell density of 0.5 × 10 ⁶ viable cells/mL in serum-free medium. A portion of the 0.5 × 10 ⁶ viable cells/mL hepatocytes was then boiled for 5 minutes and added to the plate as a negative control to eliminate enzyme activity, so that little or no substrate conversion should be observed. The boiled hepatocytes were used to prepare the negative sample. 198 μL of hepatocytes were distributed in aliquots to each well of an uncoated 96-well plate. The plate was placed in an incubator on a 500 rpm orbital shaker for approximately 10 minutes. 2 μL of 100 μM test compound or positive control were added to each well of an uncoated 96-well plate to initiate the reaction. The assay was repeated. The plate was cultured in an incubator on a 500 rpm orbital shaker for the specified time point. 25 μL of contents were removed and mixed with 6 volumes (150 μL) of cold acetonitrile containing IS (200 nM imipramine, 200 nM labetalol, and 200 nM diclofenac) to terminate the reaction at 0, 15, 30, 60, 90, and 120 minutes. The sample was centrifuged at 3220 g for 25 minutes, and 150 μL of supernatant was used for LC-MS/MS analysis. For data analysis, all calculations were performed using Microsoft Excel. Peak areas were determined from extracted ion chromatograms. The in vitro half-life (t _1/2 ) of the parent compound was determined by regression analysis of the percent disappearance of the parent compound versus time. The in vitro half-life (in vitro t _1/2 ) was determined from the slope value: in vitro t _1/2 = 0.693/k. The in vitro t _1/2 (in minutes) was converted to the scaled unbound intrinsic clearance (scaled unbound CL _int , mL/min/kg) using the following equation (mean of replicates): scaled unbound CL _int = kV/N × scaling factor, where V = culture volume (0.5 mL); N = number of hepatocytes per well (0.25×10 ⁶ cells). The scaling factor for predicting in vivo intrinsic clearance using human hepatocytes is as follows: liver weight (g liver/kg body weight): 25.7; hepatocyte concentration (10 ⁶ cells/g liver): 99; scaling factor: 2544.3.

Table 3: Human hepatocyte clearance of exemplary compounds

*: Control 1: 3-(3-(3-(2,4-difluorophenyl)ureido)-4-(diisobutylamino)phenyl)-4,4,4-trifluorobutanoic acid. See Example 30 of WO2014/150646.

Example 58: Determination of IDO inhibition in human whole blood

About 50-80mL of human blood was collected from each healthy donor in a tube containing sodium heparin. The tube containing human blood was fixed on a rotator for use. The following solutions were prepared for measurement: 10×LPS (Sigma#L2630) solution at a concentration of 1000ng/ml in RPMI culture medium (with 10mM HEPES), 10×INF-γ (INF-gamma, R&D Systems#CA31639) solution in RPMI culture medium (with 10mM HEPES), and 75× compound/inhibitor solution in 100% DMSO. The entire contents of the human blood were poured from the tube into a storage vessel, and about 120uL of blood was transferred from the vessel to each well of a 96-well plate (polypropylene U-bottom transparent plate). Then 15μL each of 10×LPS and 10×INF-γ and 2μL of 75× compound solution were added to each well. The 96-well plate was gently rotated to mix the solution and then covered with a breathable membrane. The plates were transferred to a cell culture incubator (37°C, 5% _CO2 ). After 18 hours of incubation, the plates were spun at 1800 rpm for 10 minutes without braking to separate the plasma from the blood cells. 60 μL of plasma was gently removed without disturbing the cells in each well. Kynurenine and tryptophan in the plasma were analyzed by LC-MS.

Figure 3A is a graph showing the percent inhibition of the kynurenine/tryptophan ratio by compound 84 and compound INCB-24360, respectively, as a function of the concentration of each compound. Figure 3B is a graph showing the percent inhibition of kynurenine by compound 84 and compound INCB-24360, respectively, as a function of the concentration of each compound.

Example 59: T cell and HeLa cell co-culture assay

HeLa cells are seeded into 96-well plates (5000 cells per well, in 100 μL cell growth medium DMEM containing 10% FBS and 1% penicillin/streptomycin) and cultivated at 37°C with 5% CO _2. After incubation overnight, 200 μL of INF-γ (50 ng/ml in growth medium) are added to the plate and returned to the incubator for 48 hours. According to the manufacturer's instructions (Stemcell), SepMate ^TM -50 centrifuge tubes are used to separate PBMC from human donors. EasySep human T cell isolation kit (Stemcell) is then used to separate CD3 T cells from PBMC. 96-well plates are washed twice with 200 μL co-culture medium (RPMI-1640+10% FBS+1% penicillin/streptomycin). CD3 T cell density was adjusted to 5 × ¹⁰ cells/ml using high-dose anti-CD3/CD28 beads and co-culture medium (RPMI-1640 + 10% FBS + 1% penicillin/streptomycin) and seeded into 96-well plates at 200 μL/well. Compounds of varying concentrations were added to each well, and the plates were incubated for an additional 72 hours. The level of INF-γ in the co-culture medium (100 μL) was analyzed using the human IFN-γ ELISA Ready-SET-GO kit from eBioscience. The results obtained in this example are shown in Figure 4.

Equivalents and scope

In the claims, unless indicated to the contrary or obvious from the context, articles such as "a," "an," and "the" may mean one or more than one. Unless indicated to the contrary or obvious from the context, a claim or specification that includes "or" between one or more members of a group is deemed to satisfy that one, more than one, or all of the group members are present, employed, or associated with a given product or method. The invention includes embodiments in which exactly one member of the group is present, employed, or associated with a given product or method. The invention includes embodiments in which more than one or all of the group members are present, employed, or associated with a given product or method.

In addition, the present invention encompasses all variations, combinations and arrangements of one or more limitations, elements, clauses, descriptive terms, etc., introduced into another claim from one or more listed claims. For example, any claim attached to another claim may be modified to include one or more limitations found in any other claim attached to the same basic claim. When elements are provided in an enumerated form, for example, in the form of Markush groups, each subgroup of the elements is also disclosed, and any element can be removed from the group. It should be understood that, in general, when the present invention or aspects of the present invention are described as comprising specific elements and/or features, certain embodiments of the present invention or aspects of the present invention are composed of or substantially comprised of the elements and/or features. For simplicity, those embodiments in this regard are not specifically stated herein. It should also be noted that the terms "comprising" and "including" are intended to be open and allow the inclusion of other elements or steps. When a range is given, the endpoints are included. Furthermore, unless otherwise indicated or otherwise apparent from the context and/or understanding of one skilled in the art, values expressed in range format may be assumed to be any specific value or sub-range within the specified range in different embodiments of the present invention, to the tenth of the unit of the lower limit of the range unless otherwise expressly stated herein.

This application cites various granted patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. If there is a conflict between any incorporated reference and this specification, the specification shall prevail. In addition, any specific embodiment of the present invention that is in the prior art may be explicitly excluded from any one or more claims. Because such embodiments are considered to be known to those of ordinary skill in the art, they may be excluded even if such exclusion is not explicitly stated herein. Any specific embodiment of the present invention may be excluded from any claim for any reason, regardless of whether it is relevant to the existence of the prior art.

Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments described herein. The scope of the embodiments described herein is not intended to be limited to the foregoing description, but is set forth in the appended claims. Those skilled in the art will appreciate that various changes and modifications may be made to this description without departing from the spirit or scope of the invention, as defined in the following claims.

Claims (29)

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof: in: W is the key; Q is the key; Where the oxidation state allows, Y is -CR ⁸ = or -N =. ^R1 is -C(=O)OH, -C(=O)OR ¹⁰ , or ^R2 and ^R3 are each independently hydrogen, halogen, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, or ^R2 and ^R3 are connected to form a 3- to 8-membered carbon ring or a 3- to 8-membered heterocycle; ^R4 and ^R5 are each independently a _C1 - _C6 alkyl, _C2 - _C6 alkenyl, _C5 - _C8 cycloalkenyl, _C2 - _C10 alkynyl, _C1 - _C6 alkoxy, C3- _C8 cycloalkyl, 3- to ₁₂ -membered heterocyclic, 5- to 6-membered monocyclic heteroaryl, 8- to 10-membered bicyclic heteroaryl, _C6 - _C14 aryl, or _C6 - _C14 arylsulfonyl; or ^R4 and ^R5 are connected together with the N to which they are attached to form one of the following: ^R6 is a _C1 - _C6 alkyl, _C3 - _C8 cycloalkyl, C2- _C6 alkenyl, _C2 - _C6 ynyl, _C5 - _C8 cycloalkenyl, _C6 - _C14 aryl, 4- to 7 _- membered monocyclic heterocyclic, 7- to 10-membered bicyclic heterocyclic, 5- to 6-membered monocyclic heteroaryl, or 8- to 10-membered bicyclic heteroaryl; ^R8 is hydrogen, halogen, -CN, -OH, _C1 - _C6 alkyl, or _C1 - _C6 alkoxy; and ^R10 is a _C1 - _C6 alkyl or _C2 - _C6 alkenyl. 2. The compound of claim 1, wherein Y is -CH=. 3. The compound according to any one of claims 1 to 2, wherein ^R1 is –C(＝O)OH or 4. The compound according to any one of claims 1 to 2, wherein ^R2 and ^R3 are each independently hydrogen or _C1 - _C6 alkyl. 5. The compound of claim 4, wherein ^R2 and ^R3 are each independently methyl or ethyl. 6. The compound of claim 1, wherein ^R2 and ^R3 are connected to form a 3- to 6-membered carbon ring or a 3- to 8-membered heterocycle. 7. The compound according to any one of claims 1 to 2, wherein ^R2 and ^R3 are connected to form a cyclopropyl ring, a cyclobutyl ring, a cyclopentyl ring, or a cyclohexyl ring. 8. The compound of claim 7, wherein ^R2 and ^R3 are linked to form an unsubstituted cyclopropyl ring, an unsubstituted cyclobutyl ring, or an unsubstituted cyclopentyl ring. 9. The compound of claim 6, wherein ^R2 and ^R3 are linked to form a tetrahydropyran ring. 10. The compound according to any one of claims 1 to 2, wherein ^R4 and ^R5 are each independently a _C1 - _C6 alkyl, _C3 - _C8 cycloalkyl, or a 3- to 12-membered heterocyclic group. 11. The compound of claim 10, wherein ^R4 and ^R5 are each independently of the following formula: Where R is a _C1 - _C6 alkyl group. 12. The compound of claim 10, wherein ^R4 and ^R5 are each independently of the following formula: 13. The compound of claim 10, wherein ^R4 and ^R5 are each independently of the following formula: 14. The compound according to any one of claims 1 to 2, wherein R ⁶ is selected from benzyl, benzoyl, 5 to 7-membered monocyclic heterocyclic group, 5 to 6-membered monocyclic heteroaryl group and 7 to 10-membered bicyclic heterocyclic group. 15. The compound according to any one of claims 1 to 2, wherein R ⁶ is of the following formula: Wherein R ^6A is hydrogen, C ₁ -C ₆ alkyl, halogen, -CN, -OR ^6a , or sulfonyl; Wherein R ^6a is hydrogen, or a _C1 - _C6 alkyl group; and k can be 0, 1, or 2. 16. The compound according to any one of claims 1 to 2, wherein R ⁶ has the following formula: 17. The compound according to any one of claims 1 to 2, wherein R ⁶ has the following formula: 18. The compound according to any one of claims 1 to 2, wherein R ⁶ has the following formula: 19. The compound according to any one of claims 1 to 2, wherein R ⁹ and R ¹⁰ are each independently hydrogen. 20. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is: 21. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is: 22. A pharmaceutical composition comprising the compound of any one of claims 1 to 21, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. 23. Use of the compound of any one of claims 1 to 21 or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition of claim 22, in the preparation of a medicament for treating diseases associated with IDO. 24. The use as described in claim 23, wherein the disease associated with IDO is cancer or an infectious disease. 25. The use as described in claim 23, wherein the disease associated with IDO is cancer, selected from lung cancer, breast cancer, prostate cancer, ovarian cancer, endometrial cancer, cervical cancer, bladder cancer, head and neck cancer, renal cell carcinoma, esophageal cancer, pancreatic cancer, brain cancer, gastrointestinal cancer, liver cancer, leukemia, lymphoma, melanoma, multiple myeloma, Ewing's sarcoma, and osteosarcoma. 26. The use as described in claim 23, wherein the use is in combination with an anticancer agent. 27. The use as described in claim 23, wherein the IDO-related disease is an infectious disease. 28. The use as described in claim 27, wherein the use is in combination with another antiviral agent or antiviral vaccine. 29. A reagent kit comprising: The compound of any one of claims 1 to 21, a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 22; and Instructions for use of the compound or pharmaceutical composition.