Azetidinyl-acetamides as CXCR7 inhibitors

Abstract

Formula I: TIFF2024518145000123.tif29170 or a pharma- ceutically acceptable salt, hydrate or N-oxide thereof are provided, which are useful for binding to CXCR7 and treating diseases dependent at least in part on CXCR7 activity. Thus, in a further aspect, the invention provides compositions comprising one or more of the above compounds in admixture with a pharma- ceutically acceptable excipient.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority under 35 U.S.C. §119(e) to U.S. Provisional Patent Application No. 63 / 176,451, filed April 19, 2021, the entire disclosure of which is incorporated herein by reference.

[0002] STATEMENT REGARDING RIGHTS TO INVETIONS MADE UNDER FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT Not applicable.

[0003] Reference to a "Sequence Listing," a table, or a computer program listing appendix submitted on a compact disc. Not applicable.

[0004] The present invention is directed to novel compounds and pharmaceutical compositions that inhibit the binding of the SDF-1 chemokine (also known as the CXCL12 chemokine) or I-TAC (also known as CXCL11) to the chemokine receptor CXCR7 (also known as ACKR3). These compounds are useful in preventing tumor cell proliferation, tumorigenesis, tumor angiogenesis, metastasis, inflammatory diseases including, but not limited to, arthritis, renal inflammatory disorders and multiple sclerosis, conditions of inappropriate angiogenesis including, but not limited to, wound healing, treatment of HIV infection, and treatment of stem cell differentiation and mobilization disorders, acute renal failure, hemolytic uremic syndrome, ischemia-reperfusion injury, opioid addiction, and neuropathic pain (see also co-pending U.S. Patent Application Publication Nos. 10 / 912,638, 11 / 407,729, and 11 / 050,345). [Background technology]

[0005] Chemokines are a superfamily of small cytokine-like proteins that can induce cytoskeletal rearrangements, firm adhesion of leukocytes to endothelial cells, degranulation and directional migration of leukocytes, and result in cell activation and proliferation. Chemokines act in concert with cell surface proteins to induce specific homing of various subsets of cells to specific anatomical sites.

[0006] Early research efforts by many groups have demonstrated the role of the chemokine receptor CXCR4 in metastasis and tumor growth. Muller, et al., "Involvement of Chemokine Receptors in Breast Cancer Metastasis," Nature, 410:50-56 (2001) demonstrated that breast cancer cells use chemokine-mediated mechanisms, such as those regulating leukocyte trafficking, in the process of metastasis. Tumor cells express distinct, nonrandom patterns of functionally active chemokine receptors. Signaling through CXCR4 mediates actin polymerization and pseudopodia formation in breast cancer cells, inducing chemotactic and invasive responses. Furthermore, organs that represent the primary sites of breast cancer metastasis (e.g., lymph nodes, bone marrow, and lungs) are the most abundant sources of ligands for the CXCR4 receptor.

[0007] Using immunodeficient mice, Muller et al. were able to reduce metastasis of injected human breast cancer cells by treating the mice with an antibody known to bind CXCR4, a finding that suggests that breast cancer metastasis may be reduced by treating patients with CXCR4 antagonists.

[0008] Bertolini, et al., "CXCR4 Neutralization, a Novel Therapeutic Approach for Non-Hodgkin's Lymphoma," Cancer Research, 62:3106-3112 (2002) demonstrated reduced tumor volume and increased survival in immunodeficient mice injected with human lymphoma cells treated with anti-CXCR4 antibodies. Bertolini et al. interpreted their findings to mean that tumor volume could be reduced by treating patients with CXCR4 antagonists.

[0009] More recent studies have suggested that another chemokine receptor, CXCR7, may also be a target in cancer therapy. CXCR7 is preferentially expressed in transformed cells over normal cells, and expression is detectable in many human cancers. In vitro studies show that proliferation of CXCR7-expressing cells can be inhibited by antagonists of CXCR7. In vivo studies in mice show that CXCR7 antagonists can inhibit tumor formation and tumor growth (reviewed in Morian, D. et al., Front Immunol (2020) 11:952-971).

[0010] Specific CXCR7 antagonists can prevent the growth and spread of cancer, and expression patterns show a restricted tissue distribution of the CXCR7 receptor that correlates with tumor formation (Luo, Y. et al., Int. J. Cancer, (2018) 142: 2163-2174).

[0011] Additionally, CXCR7 can serve as a coreceptor for certain genetically divergent human immunodeficiency viruses (HIV) and simian immunodeficiency viruses (SIV), particularly HIV-2-ROD, an X4-tropic isolate (Shimizu, N. et al., J. Virol., (2000) 74:619-626; Balabian, K., et al., J. Biol. Chem., (2005) 280:35760-35766; D'huys, T., et al., Heliyon, (2018) 4:e00557).

[0012] Furthermore, SDF-1 has been described to have a role in the mobilization of hematopoietic progenitor and stem cells, in particular the mobilization of cells bearing the CXCR4 receptor from certain hematopoietic tissues, including bone marrow (Hattori, K., et al., Blood, (2000) 97:3354-3360; WO 2005 / 000333, the disclosure of which is incorporated herein by reference). More recent studies suggest that the CXCR7 receptor may also play a role in stem cell mobilization processes (Melo, RD et al., Stem Cell Res Ther, (2018) 9:34-38).

[0013] In addition to the original discovery of the role of CXCR7 in cancer and stem cell mobilization, modulating this receptor has shown promise in preventing HIV infection, Shiga toxin-associated hemolytic uremic syndrome, ischemia-reperfusion injury in cardiac tissue and kidney transplants, and reducing myosclerotic lesions. The recent discovery that CXCR7 binds opioids suggests that modulating this receptor could potentially modulate pain and treat opioid addiction. [Prior art documents] [Patent documents]

[0014] [Patent Document 1] US Patent Application Publication No. 10 / 912,638 [Patent Document 2] US Patent Application Publication No. 11 / 407,729 [Patent Document 3] U.S. Patent Application Publication No. 11 / 050,345 [Patent Document 4] International Publication No. 2005 / 000333 Brochure [Non-patent literature]

[0015] [Non-Patent Document 1] Muller, et al., “Involvement of Chemokine Receptors in Breast Cancer Metastasis,” Nature, 410:50 - 56(2001) [Non - Patent Document 2] Bertolini, et al., “CXCR4 Neutralization, a Novel Therapeutic Approach for NonHodgkin’s Lymphoma,” Cancer Research, 62:3106 - 3112(2002) [Non - Patent Document 3] Morian, D. et al., Front Immunol(2020)11:952 - 971 [Non - Patent Document 4] Luo, Y. et al., Int.J.Cancer, (2018)142:2163 - 2174 [Non - Patent Document 5] Shimizu, N. et al., J.Virol., (2000)74:619 - 626 [Non - Patent Document 6] Balabanian, K., et al., J.Biol.Chem., (2005)280:35760 - 35766 [Non - Patent Document 7] D’huys, T., et al., Heliyon, (2018)4:e00557 [Non - Patent Document 8] Hattori, K., et al., Blood, (2000)97:3354 - 3360 [Non - Patent Document 9] Melo, RD et al., Stem Cell Res Ther, (2018)9:34 - 38 [Summary of the Invention] [Means for Solving the Problems]

[0016] In view of the above, it is apparent that compounds capable of specific binding to the CXCR7 receptor are useful in the treatment of diseases and other biological conditions that may benefit from such an interaction. The present invention provides such compounds, as well as pharmaceutical compositions and related therapeutic methods.

[0017] The present invention, in one aspect, comprises a compound of formula I [ka] or a pharma- ceutically acceptable salt, hydrate, or N-oxide thereof. 1 , R 2 , R 3 , R 4 , R 4a , R 5 , R 5a , R 6 , R 7 , Ar 1 , Ar 2 , HAr and subscripts m, n, p and q) are described in the detailed description of the invention.

[0018] The compounds provided herein are useful for treating diseases that depend at least in part on CXCR7 binding and CXCR7 activity. Thus, in a further aspect, the invention provides compositions comprising one or more of the above compounds in admixture with a pharma- ceutically acceptable excipient.

[0019] In yet another aspect, the present invention provides methods of treating various diseases discussed in more detail herein, comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound of the formula above for a period of time sufficient to treat the disease.

[0020] In yet another aspect, the present invention provides methods for diagnosing disease in an individual. In these methods, a compound provided herein is administered to a subject in a labeled form, followed by imaging to determine the presence or absence of CXCR7. In a related aspect, the method for diagnosing disease is carried out by contacting a tissue or blood sample with a labeled compound provided herein, and determining the presence, absence, or amount of CXCR7 in the sample.

[0021] In some embodiments, an amount of a chemotherapeutic agent or radiation is administered to the subject before, after, or in combination with a compound of the invention, in some embodiments, when only a chemotherapeutic agent or radiation is administered, the amount is sub-therapeutic.

[0022] BRIEF DESCRIPTION OF THE DRAWINGS Not applicable. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0023] I. Abbreviations and Definitions Unless otherwise stated, the term "alkyl," by itself or as part of another substituent, means a straight or branched chain hydrocarbon group having the specified number of carbon atoms (i.e., C1-8 means 1 to 8 carbons). Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. The term "alkenyl" refers to unsaturated alkyl groups having one or more double bonds. Similarly, the term "alkynyl" refers to unsaturated alkyl groups having one or more triple bonds. Examples of such unsaturated alkyl groups include vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and the higher homologs and isomers.

[0024] The term "cycloalkyl" refers to a saturated or partially unsaturated hydrocarbon ring having the indicated number of ring atoms (e.g., C3~6 Cycloalkyl). Cycloalkyl is C 3~6 , C 4~6 , C 5~6 , C 3~8 , C 4~8 , C 5~8 , C 6~8 , C 3~9 and C 3~10 It can contain any number of carbons, such as cycloalkyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclooctyl. Partially unsaturated cycloalkyl groups have one or more double or triple bonds in the ring, but the cycloalkyl group is not aromatic. Saturated monocyclic cycloalkyl rings include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclooctyl.

[0025] The term "cycloalkyloxy" refers to a cycloalkyl group having an oxygen atom connecting the cycloalkyl group to the point of attachment: cycloalkyl-O-. Cycloalkyl groups are as defined herein.

[0026] The term "bridged cyclyl" or "bridged cycloalkyl" refers to a ring in which two non-adjacent ring atoms are linked together (CRR'). n R refers to cycloalkyl rings (having 4-8 ring vertices) linked by groups, where n is 1-3, and each R is independently H or methyl (which may also be referred to herein as a "bridged" group). A bridged cycloalkyl group does not have any heteroatoms as ring vertices. Additionally, C 5~8 refers to a bridged cycloalkyl group having 5 to 8 ring members. Examples include, but are not limited to, bicyclo[1.1.1]pentane, bicyclo[2.2.2]octane, bicyclo[2.2.1]heptane, and the like.

[0027] The term "spirocyclyl" or "spirocycloalkyl" refers to a saturated or partially unsaturated bicyclic ring having 6-12 ring atoms, with the two rings being linked through a single carbon atom (aka a spiro atom). Partially unsaturated spirocycloalkyl groups have one or more double or triple bonds in the ring, but the spirocycloalkyl group is not aromatic. Representative examples include, but are not limited to, spiro[3.3]heptane, spiro[4.4]nonane, spiro[3.4]octane, and the like.

[0028] The term "heterocycloalkyl" refers to a saturated or partially unsaturated monocyclic ring having the indicated number of ring vertices (e.g., 3-7 membered ring) and having 1-5 heteroatoms selected from N, O, and S as ring vertices. Partially unsaturated heterocycloalkyl groups have one or more double or triple bonds in the ring, but the heterocycloalkyl group is not aromatic. Heterocycloalkyl groups can contain any number of ring atoms, for example, 3-6, 4-6, 5-6, 3-7, 4-7, or 5-7 ring members. Any suitable number of heteroatoms can be included in the heterocycloalkyl group, such as 1, 2, 3, or 4, or 1-2, 1-3, 1-4, 2-3, 2-4, or 3-4. Non-limiting examples of heterocycloalkyl groups include pyrrolidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrahydrothiophene, quinuclidine, etc. Heterocycloalkyl groups can be attached to the remainder of the molecule through a ring carbon or heteroatom.

[0029] The term "bicyclic heterocycloalkyl" or "bicyclic heterocyclyl" refers to a saturated or partially unsaturated fused bicyclic ring having the indicated number of ring vertices (e.g., 6-12 membered ring) and having 1-5 heteroatoms selected from N, O, and S as ring vertices. Partially unsaturated bicyclic heterocycloalkyl groups have one or more double or triple bonds in the ring, but the bicyclic heterocycloalkyl group is not aromatic. Bicyclic heterocycloalkyl groups can contain any number of ring atoms, such as 6-8, 6-9, 6-10, 6-11, or 6-12 ring members. Any suitable number of heteroatoms can be included in the heterocycloalkyl group, such as 1, 2, 3, or 4, or 1-2, 1-3, 1-4, 2-3, 2-4, or 3-4. Non-limiting examples of bicyclic heterocycloalkyl groups include decahydro-1,5-naphthyridine, octahydropyrrolo[1,2-a]pyrazine, and the like.

[0030] The term "bridged heterocyclyl" or "bridged heterocycloalkyl" refers to a heterocyclic ring system in which two non-adjacent ring atoms are linked together (CRR'). n R refers to heterocycloalkyl rings (having 5-7 ring vertices) linked by a C-R group, where n is 1-3, and each R is independently H or methyl (which may also be referred to herein as a "bridged" group). Bridged heterocyclyl groups have 1-5 heteroatoms selected from N, O, and S as ring vertices. The heteroatom ring vertex may be within the heterocycloalkyl ring portion and simultaneously within the bridging group. When within the bridging group, the heteroatom replaces a CRR' group. Examples include, but are not limited to, 2-azabicyclo[2.2.2]octane, quinuclidine, 7-oxabicyclo[2.2.1]heptane, and the like.

[0031] The term "spiroheterocyclyl" or "spiroheterocycloalkyl" refers to a saturated or partially unsaturated bicyclic ring having 6 to 12 ring atoms, with the two rings connected through a single carbon atom (also known as a spiro atom). Spiroheterocyclyl groups have 1 to 5 heteroatoms selected from N, O, and S as ring vertices, with the nitrogen atom optionally being quaternized. Partially unsaturated spiroheterocycloalkyl groups have one or more double or triple bonds in the ring, but spiroheterocycloalkyl groups are not aromatic. Representative examples include, but are not limited to, 2,6-diazaspiro[3.3]heptane, 2,6-diazaspiro[3.4]octane, 2-azaspiro[3.4]octane, 2-azaspiro[3.5]-nonane, 2,7-diazaspiro[4.4]nonane, and the like.

[0032] The term "alkylene" by itself or as part of another substituent means a divalent group derived from an alkane, as exemplified by -CH2CH2CH2CH2-. Typically, an alkyl (or alkylene) group will have from 1 to 24 carbon atoms, with groups having 10 or fewer carbon atoms being preferred in the present invention. A "lower alkyl" or "lower alkylene" is a shorter chain alkyl or alkylene group, generally having four or fewer carbon atoms. Similarly, "alkenylene" and "alkynylene" refer to the unsaturated forms of "alkylene" having a double or triple bond, respectively.

[0033] As used herein, in any chemical structure depicted herein, a wavy line "" crossing a single bond, double bond, or triple bond is [ka] " represents a point attachment of a single, double or triple bond to the remainder of the molecule.

[0034] The terms "alkoxy," "alkylamino," and "alkylthio" (or thioalkoxy) are used in their conventional sense to refer to an alkyl group attached to the remainder of the molecule via an oxygen atom, an amino group, or a sulfur atom, respectively. Additionally, for dialkylamino groups, the alkyl portions may be the same or different and may also form a 3- to 7-membered ring in combination with the nitrogen atom to which each is attached. Thus, -NR a R b A group represented as: is meant to include piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl, and the like.

[0035] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as "haloalkyl" are meant to include monohaloalkyl and polyhaloalkyl. For example, "C 1~4 The term "haloalkyl" is meant to include trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0036] The term "hydroxyalkyl" is meant to refer to an alkyl group, as defined above, having one or two hydroxyl groups as substituents. For example, "C 1~6 The term "hydroxyalkyl" is meant to include 2-hydroxyethyl and 2,4-dihydroxybutyl.

[0037] The term "aryl" refers to a polyunsaturated, typically aromatic, hydrocarbon group that may be a single ring or multiple rings (up to three rings) that are fused or covalently linked together, unless otherwise stated. The term "heteroaryl" ring refers to an aryl group (or aryl ring) that contains from one to five heteroatoms selected from N, O, and S, where the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom is optionally quaternized. A heteroaryl group can be attached to the remainder of the molecule through a heteroatom. Non-limiting examples of aryl groups include phenyl, naphthyl, and biphenyl, while non-limiting examples of heteroaryl groups include pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, triazinyl, quinolinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, benzotriazinyl, purinyl, benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl, isobenzofuryl, isoindolyl, Examples include indolizinyl, benzotriazinyl, thienopyridinyl, thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridine, benzothiaxolyl, benzofuranyl, benzothienyl, indolyl, quinolyl, isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl, thiazolyl, furyl, thienyl, etc. Substituents for each of the above noted aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below.

[0038] The term "arylalkyl" is meant to include radicals in which an aryl group is attached to an alkyl group (e.g., benzyl, phenethyl, etc.). Similarly, the term "heteroaryl-alkyl" is meant to include radicals in which a heteroaryl group is attached to an alkyl group (e.g., pyridylmethyl, thiazolylethyl, etc.).

[0039] The above terms (e.g., "alkyl," "aryl" and "heteroaryl"), in some embodiments, include both substituted and unsubstituted forms of the indicated radical. Preferred substituents for each type of radical are provided below.

[0040] Substituents for alkyl radicals (including groups often referred to as alkylene, alkenyl, alkynyl, and cycloalkyl) can be a variety of groups selected from -halogen, -OR', -NR'R'', -SR', -SiR'R''R''', -OC(O)R', -C(O)R', -COR', -CONR'R'', -OC(O)NR'R'', -NR''C(O)R', -NR'C(O)NR''R'''', -NR''C(O)2R', -NHC(NH2)=NH, -NR'C(NH2)=NH, -NHC(NH2)=NR', -S(O)R', -S(O)2R', -S(O)2NR'R'', -NR'S(O)2R'', -CN, and -NO2 in numbers ranging from zero to (2m'+1), where m' is the total number of carbon atoms in such radical. R', R'' and R''' are each independently hydrogen, unsubstituted C 1~8 Alkyl, unsubstituted aryl, aryl substituted with 1 to 3 halogens, unsubstituted C 1~8 Alkyl, C 1~8 Alkoxy or C 1~8 Thioalkoxy group or unsubstituted aryl-C 1~4 It refers to an alkyl group. When R' and R" are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring. For example, -NR'R" is meant to include 1-pyrrolidinyl and 4-morpholinyl.

[0041] Similarly, substituents for the aryl and heteroaryl groups are varied and are generally selected from -halogen, -OR', -OC(O)R', -NR'R'', -SR', -R', -CN, -NO2, -CO2R', -CONR'R'', -C(O)R', -OC(O)NR'R'', -NR''C(O)R', -NR''C(O)2R', -NR'C(O)NR''R''', -NHC(NH2)=NH, -NR'C(NH2)=NH, -NHC(NH2)=NR', -S(O)R', -S(O)2R', -S(O)2NR'R'', -NR'S(O)2R'', -N3, perfluoro(C1-C4)alkoxy, and (C1-C4)alkyl, where R', R'' and R''' are independently selected from hydrogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 3~6 Cycloalkyl, C 2~8 Alkenyl, C 2~8 Selected from alkynyl, unsubstituted aryl and heteroaryl, (unsubstituted aryl)-C1-4 alkyl and unsubstituted aryloxy-C1-4 alkyl. Other suitable substituents include each of the above aryl substituents attached to a ring atom by an alkylene tether of 1 to 4 carbon atoms.

[0042] Two of the substituents on adjacent atoms of the aryl or heteroaryl ring are optionally represented by the formula -TC(O)-(CH2) q -U-, where T and U are independently -NH-, -O-, -CH2- or a single bond, and q is an integer from 0 to 2. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula A-(CH2) r A and B may be replaced with a substituent of formula -B-, where A and B are independently -CH2-, -O-, -NH-, -S-, -S(O)-, -S(O)2-, -S(O)2NR'- or a single bond, and r is an integer from 1 to 3. One of the single bonds of the new ring thus formed may optionally be replaced with a double bond. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of formula -(CH2)s -X-(CH2) t where s and t are independently integers from 0 to 3, and X is -O-, -NR'-, -S-, -S(O)-, -S(O)2-, or -S(O)2NR'-. The substituent R' in -NR'- and -S(O)2NR'- is selected from hydrogen or unsubstituted C1-6 alkyl.

[0043] As used herein, the term "heteroatom" is meant to include oxygen (O), nitrogen (N), sulfur (S) and silicon (Si).

[0044] The term "pharmaceutical acceptable salts" is intended to include salts of active compounds prepared with relatively non-toxic acids or bases, depending on the specific substituents contained in the compounds described herein. When the compounds of the present invention contain relatively highly acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutical acceptable salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, zinc, and the like. Salts derived from pharma- ceutically acceptable organic bases include salts of primary, secondary, and tertiary amines, including substituted, cyclic, and naturally occurring amines, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like. When a compound of the present invention contains a relatively highly basic functional group, an acid addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired acid, neat or in a suitable inert solvent. Examples of pharma- ceutically acceptable acid addition salts include those derived from inorganic acids, such as the hydrochloride, hydrobromide, nitrate, carbonate, monohydrogencarbonate, phosphate, monohydrogenphosphate, dihydrogenphosphate, sulfate, monohydrogensulfate, hydroiodide or phosphite salts, and the like, as well as salts derived from relatively non-toxic organic acids, such as acetate, propionate, isobutyrate, malonate, benzoate, succinate, suberate, fumarate, mandelate, phthalate, benzenesulfonate, p-tolylsulfonate, citrate, tartrate and methanesulfonate salts.Also included are salts of amino acids, such as arginates, and salts of organic acids, such as glucuronic acid or galacturonic acid (see, e.g., Berge, SM, et al, "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19).Certain compounds of the present invention contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0045] The neutral forms of the compounds can be regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but otherwise is equivalent to the parent form of the compound for purposes of the present invention.

[0046] In addition to salt forms, the present invention provides compounds that are in prodrug form. Prodrugs of the compounds described herein are compounds that readily undergo chemical changes under physiological conditions to become the compounds of the present invention. In addition, prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to the compounds of the present invention when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

[0047] Certain compounds of the present invention can exist in solvated forms, including unsolvated forms and hydrated forms.In general, solvated forms are equivalent to unsolvated forms and are intended to be included within the scope of the present invention.Certain compounds of the present invention can exist in multiple crystalline or amorphous forms.In general, all physical forms are equivalent for the use contemplated by the present invention and are intended to be within the scope of the present invention.

[0048] Certain compounds of the present invention have asymmetric carbon atoms (optical centers) or double bonds, and all racemates, diastereomers, geometric isomers, positional isomers, and individual isomers (e.g., separated enantiomers) are intended to be encompassed within the scope of the present invention. In some embodiments, the compounds of the present invention exist in enantiomerically enriched forms, and the enantiomeric excess for a particular enantiomer is calculated by known methods. The preparation of enantiomerically enriched forms is also well known in the art, and can be achieved, for example, using chiral resolution via chromatography or chiral salt formation. Furthermore, various conformers are contemplated by the present invention, as well as various rotamers. Conformers are stereoisomers that may differ in rotation about one or more σ-bonds. Rotamers are conformers that differ in rotation about only a single σ-bond. Still further, the compounds of the present invention may also contain unnatural proportions of isotopes at one or more of the atoms that constitute such compounds. Thus, in some embodiments, the compounds of the invention are present in isotopically enriched form. The non-natural proportion of an isotope may be defined as a range from the amount found in nature of the atom in question to an amount consisting of 100%. For example, the compounds may contain, for example, tritium ( 3 H), iodine-125( 125 I) or carbon-14 ( 14 Radioisotopes such as C or deuterium ( 2 H) or carbon-13 ( 13 Non-radioactive isotopes such as C) may be incorporated. Such isotopic variations may provide additional utility to those described elsewhere in this application. For example, isotopic variants of the compounds of the invention may find additional utility, including but not limited to, as diagnostic and / or imaging reagents or cytotoxic / radiotoxic therapeutic agents. Furthermore, isotopic variants of the compounds of the invention may have altered pharmacokinetic and pharmacodynamic properties that may contribute to increased safety, tolerability, or efficacy during treatment. All isotopic variations of the compounds of the invention, whether radioactive or not, are intended to be encompassed within the scope of the invention.

[0049] "CXCR7" is also referred to as "RDC1" or "CCXCKR2" refers to a seven-transmembrane domain putative G-protein coupled receptor (GPCR). The CXCR7 canine orthologue was first identified in 1991. See Libert et al. Science 244:569-572 (1989). The canine sequence is described in Libert et al., Nuc. Acids Res. 18(7):1917 (1990). The mouse sequence is described, for example, in Heesen et al., Immunogenetics 47:364-370 (1998). The human sequence is described, for example, in Sreedharan et al., Proc. Natl. Acad. Sci. USA 88:4986-4990 (1991), which erroneously describes the protein as a receptor for vasoactive intestinal peptide.

[0050] II. Overview The compounds of the present invention can inhibit the binding of ligands to the CXCR7 receptor and are useful for treating various diseases, such as cancer, particularly solid tumor cancer and lymphoma.Recently, it has been found that the inhibition of ligand binding to CXCR7 reduces the severity of rheumatoid arthritis in animal models.

[0051] Those skilled in the art will understand that agents that modulate CCX-CKR2 activity (CXCR7 activity) can be combined with other anti-angiogenic agents and / or chemotherapeutic agents or radiation and / or other anti-arthritic agents in a treatment regimen. In some cases, the amount of chemotherapeutic agent or radiation is sub-therapeutic when provided without combination with an anti-angiogenic agent. Those skilled in the art will understand that "combination" can include combination in a treatment (i.e., two or more drugs are administered as a mixture or at least simultaneously, or at least introduced into a subject at different times, but both are introduced so that they are simultaneously present in the subject's bloodstream). Furthermore, the compositions of the present invention can be administered before or after a second treatment regimen, such as before or after administration of chemotherapy or radiation therapy.

[0052] III. EMBODIMENTS OF THE PRESENTINVENTION A. Compound The present invention, in one aspect, comprises a compound of formula I [ka] or a pharma- ceutically acceptable salt, hydrate, N-oxide, isotopically or enantiomerically enriched version, or rotamer thereof, wherein HAr is a 5-membered heteroaryl ring; Ar 1 is selected from the group consisting of phenyl, pyridyl, pyrimidinyl and pyrazinyl; Ar 2 are each independently a monocyclic or fused bicyclic aryl or heteroaryl; The subscript m is 0, 1 or 2; The subscript n is 0, 1, 2 or 3; the subscript p is 0, 1, 2 or 3; the subscript q is 0, 1, 2, 3 or 4; Each R 1 are independently halogen, CN, C 1~4 Alkyl, C 1~4 Haloalkyl, -NR a R b , -OR a , -CO2R a and -C(O)NR a R b is a member selected from the group consisting of Each R 2 are independently halogen, CN, C 1~4 Alkyl, C 1~4 Haloalkyl, -NR a R b , -OR a , -CO2R a and -C(O)NR a R b is a member selected from the group consisting of Each R 3 is C 1~4 Alkyl, C 1~4 Haloalkyl, C 1~4Hydroxyalkyl, -CO2R a , -X-CO2R a , -C(O)NR a R b and -XC(O)NR a R b is a member selected from the group consisting of R 4a and R 5a Each of the groups is independently H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Hydroxyalkyl, -X-OR a , -CO2R a , -X-CO2R a , -X-NR a R b , -C(O)NR a R b and -XC(O)NR a R b is a member selected from the group consisting of R 4 and R 5 Each of the groups is independently H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Hydroxyalkyl, -X-OR a , -CO2R a , -X-CO2R a , -X-NR a R b , -C(O)NR a R b and -XC(O)NR a R b or R 4 and R 5 are combined to form a 3- to 5-membered ring having 0 or 1 heteroatom ring vertex selected from O, S, or N, the 3- to 5-membered ring being unsubstituted or substituted with halogen, CN, C 1~4 Alkyl, C 1~4 Haloalkyl, C 1~4 Alkoxy and C 1~4 substituted with 1 to 4 substituents independently selected from the group consisting of haloalkoxy; Each R 6are independently halogen, CN, -X-CN, C 1~4 Alkyl, C 1~4 Haloalkyl, C 3~6 Cycloalkyl, C 1~4 Hydroxyalkyl, -OR a , -CO2R a , -X-CO2R a , -NR a R b , -X-NR a R b , -C(O)NR a R b and -XC(O)NR a R b is a member selected from the group consisting of Each R 7 is C 1~8 Alkyl, C 3~8 Hydroxyalkyl, C 1~4 Alkoxy-C 2~4 Alkyl, -C(O)NH-C 1~8 Alkyl, -C(O)-C 1~8 Alkyl, -S(O)2-C 1~8 Alkyl, C 3~8 Cycloalkyl, -XC 3~8 Cycloalkyl, C 6~9 Spirocycloalkyl, -XC 6~9 is a member selected from the group consisting of spirocycloalkyl, 4- to 7-membered heterocycloalkyl, -X-4- to 7-membered heterocycloalkyl, 7- to 11-membered spiroheterocycloalkyl, and -X-7- to 11-membered spiroheterocycloalkyl, 7 is substituted with 0 to 4 substituents independently selected from the group consisting of hydroxy, methyl, ethyl, hydroxymethyl, fluoro, chloro, methoxy, ethoxy, and cyclopropyl; Each R a and R b are independently H, C 1~4 Alkyl, C 1~4 Haloalkyl, C 3~6 Cycloalkyl and C 3~6 Cycloalkyl-C 1~4 is selected from the group consisting of alkyl, Each X is C 1~4is an alkylene linking group, any of the methylene moieties of X is unsubstituted or substituted with one or two methyl groups.

[0053] In one group of embodiments, compounds having formula I are provided, wherein HAr is selected from the group consisting of isoxazole, isothiazole, imidazole, pyrazole, thiazole, 1,2,4-oxadiazole, 1,3,4-oxadiazole, 1,2,4-thiadiazole, 1,3,4-thiadiazole, 1,2,3-triazole, and 1,2,4-triazole. In another group of embodiments, HAr is selected from isoxazole and thiadiazole.

[0054] In one group of embodiments, compounds are provided having formula I, wherein Ar 1 is phenyl.

[0055] In one group of embodiments, in addition to the additional embodiments above, there are provided compounds having formula I, wherein subscript q is 1, 2, or 3, and each R 1 are independently halogen, CN, C 1~4 Alkyl and C 1~4 haloalkyl.

[0056] In another group of embodiments, in addition to the additional embodiments above, there are provided compounds having Formula I, wherein Ar 2 is selected from the group consisting of pyridyl, pyrimidinyl, pyrazinyl, phenyl, indolyl, thiazolyl, pyrazolyl, indazolyl, and pyrrolopyridinyl. 2 is selected from pyrimidinyl, pyridyl and phenyl.

[0057] In another group of embodiments, in addition to the additional embodiments above, there are provided compounds having Formula I, wherein Ar 2 is selected from the group consisting of 2-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 2-thiazolyl, 4-pyrazolyl, phenyl, and indolyl, and each R 6are independently halogen, CN, C 1~4 Alkyl, C 1~4 Haloalkyl, C 3~6 Cycloalkyl and C 1~4 alkoxy is selected from the group consisting of

[0058] In another group of embodiments, in addition to the additional embodiments above, there are provided compounds having Formula I, wherein -Ar 2 -(R 6 ) p is selected from the group consisting of: [ka]

[0059] In another group of embodiments, in addition to the additional embodiments above, there are provided compounds having formula I, wherein R 7 is selected from the group consisting of: [ka]

[0060] In another group of embodiments, further to the additional embodiments above, there are provided compounds having Formula I, wherein subscript m is 0.

[0061] In another group of embodiments, further to the additional embodiments above, there are provided compounds having formula I, wherein subscript n is 0.

[0062] In another group of embodiments, further to the additional embodiments above, there are provided compounds having formula I, wherein subscript p is 0, 1 or 2.

[0063] In another group of embodiments, further to the additional embodiments above, there are provided compounds having formula I, wherein subscript q is 1 or 2.

[0064] In another group of embodiments, the compound of formula (Ia): [ka] or a pharma- ceutically acceptable salt thereof, wherein the variables have the meanings provided in Formula I or any of the above embodiments. In some selected embodiments, HAr is isoxazole or thiadiazole.

[0065] In another group of embodiments, the compound of formula (Ia1): [ka] or a pharma- ceutically acceptable salt thereof, wherein the variables have the meanings provided in Formula I or any of the above embodiments. In some selected embodiments, HAr is isoxazole or thiadiazole.

[0066] In another group of embodiments, the compound of formula (Ia2): [ka] or a pharma- ceutically acceptable salt thereof, wherein the variables have the meanings provided in Formula I or any of the above embodiments. In some selected embodiments, HAr is isoxazole or thiadiazole.

[0067] In another group of embodiments, compounds of formula (Ib), (Ic) or (Id): [ka] or a pharma- ceutically acceptable salt thereof, wherein the variables have the meanings provided in Formula I or any of the above embodiments. In some selected embodiments, HAr is isoxazole or thiadiazole.

[0068] In another group of embodiments, compounds of formula (Ib1), (Ic1) or (Id1): [ka] or a pharma- ceutically acceptable salt thereof, wherein the variables have the meanings provided in Formula I or any of the above embodiments. In some selected embodiments, HAr is isoxazole or thiadiazole.

[0069] In another group of embodiments, in addition to the additional formulas and embodiments above, there are provided compounds having formula I, wherein R 7 is C 1~8 Alkyl, C 3~8 Hydroxyalkyl, C 1~4 Alkoxy-C 2~4 Alkyl, C 3~8 Cycloalkyl, C 6~9 is a member selected from the group consisting of spirocycloalkyl, 4- to 7-membered heterocycloalkyl, and 7- to 11-membered spiroheterocycloalkyl; 7 is substituted with 0 to 4 substituents independently selected from the group consisting of hydroxy, methyl, ethyl, hydroxymethyl, fluoro, chloro, methoxy, ethoxy, and cyclopropyl.

[0070] In another group of embodiments, in addition to the additional formulas and embodiments above, there are provided compounds having formula I, wherein R 7 -XC 3~8 Cycloalkyl, -XC 6~9 is a member selected from the group consisting of spirocycloalkyl, -X-4 to 7 membered heterocycloalkyl, and -X-7 to 11 membered spiroheterocycloalkyl, 7 is substituted with 0 to 4 substituents independently selected from the group consisting of hydroxy, methyl, ethyl, hydroxymethyl, fluoro, chloro, methoxy, ethoxy, and cyclopropyl.

[0071] In another group of embodiments, in addition to the additional formulas and embodiments above, there are provided compounds having formula I, wherein R 7is a member selected from the group consisting of cyclohexyl, cyclopentyl, piperidinyl, tetrahydropyranyl, and tetrahydrofuranyl, each substituted with 0-2 substituents independently selected from the group consisting of hydroxy, methyl, ethyl, hydroxymethyl, fluoro, chloro, methoxy, and ethoxy.

[0072] In one selected group of embodiments, the compound is selected from those listed in the Examples or Table 1 below.

[0073] In other selected embodiments, the described compounds may be present in a pharma- ceutically acceptable salt or hydrate form.

[0074] Still further, with respect to the compounds shown above without stereochemistry, the present invention also covers chiral forms of each compound and enantiomerically enriched forms of the described compounds. Enantiomerically enriched forms can be prepared using chiral chromatography according to well-known methods practiced in the art or by chiral resolution, for example, using chiral salt forms. In some embodiments, the enantiomeric excess of the enantiomerically enriched form is at least 10%, 20%, 30%, 40%, 50%, 60% or more. In still other embodiments, enantiomerically enriched forms of at least 70%, 80%, 90%, 95% or more are provided.

[0075] Preparation of compounds The compounds of the invention can be prepared according to the methods described in the Examples section herein. In addition, the synthesis of certain intermediate compounds useful in the preparation of the compounds of the invention is also described.

[0076] B. Composition In addition to the compounds provided above, compositions for modulating CXCR7 activity in humans and animals will usually contain a pharmaceutical carrier or diluent.

[0077] The term "composition," as used herein, is intended to encompass any product containing the specified ingredients in the specified amounts, and any product that results directly or indirectly from the combination of the specified ingredients in the specified amounts. "Pharmaceutically acceptable" means the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

[0078] The pharmaceutical composition for administering the compound of the present invention can be conveniently provided in unit dosage form and can be prepared by any method known in the art of pharmacy and drug delivery.All methods include the step of associating the active ingredient with the carrier that constitutes one or more auxiliary ingredients.In general, the pharmaceutical composition can be prepared by homogenizing and finely dividing the active ingredient with liquid carriers or finely divided solid carriers or both, and then shaping the product into desired formulations as necessary.In this pharmaceutical composition, the active target compound is included in an amount sufficient to produce the desired effect depending on the process or condition of the disease.

[0079] The pharmaceutical composition containing the active ingredient may be in a form suitable for oral use, such as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions and self-emulsifying formulations as described in US Patent Publication No. 2002 / 0012680, hard or soft capsules, syrups, elixirs, liquids, buccal patches, oral gels, chewing gums, chewable tablets, effervescent powders and effervescent tablets. Compositions intended for oral use may be prepared by any method known in the art for producing pharmaceutical compositions, and such compositions may contain one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents, antioxidants and preservatives to provide pharma-ceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharma-ceutically acceptable excipients suitable for the manufacture of tablets. These excipients can be, for example, inert diluents such as cellulose, silicon dioxide, aluminum oxide, calcium carbonate, sodium carbonate, glucose, mannitol, sorbitol, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents such as corn starch or alginic acid; binders such as PVP, cellulose, PEG, starch, gelatin or acacia; and lubricants such as magnesium stearate, stearic acid or talc. The tablets can be uncoated or enteric or otherwise coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a longer period of time. For example, a time-delay material such as glyceryl monostearate or glyceryl distearate can be used. They can also be coated by the techniques described in U.S. Pat. Nos. 4,256,108, 4,166,452 and 4,265,874 to form osmotic therapeutic tablets for controlled release.

[0080] Formulations for oral use may also be provided as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin or olive oil.Furthermore, emulsions may be prepared with non-water-miscible ingredients, such as oils, and stabilized with surfactants, such as mono- or diglycerides, PEG esters, etc.

[0081] Aqueous suspensions contain the active substance mixed with excipients suitable for the manufacture of aqueous suspensions.Such excipients are suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia;dispersing agents or wetting agents can be natural phosphatides, such as lecithin, or condensation products of alkylene oxides with fatty acids, such as polyoxyethylene stearate, or condensation products of ethylene oxide with long-chain aliphatic alcohols, such as heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with fatty acids and partial esters derived from hexitols, such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with fatty acids and partial esters derived from hexitol anhydrides, such as polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives, for example ethyl or n-propyl, p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.

[0082] Oily suspensions can be formulated by suspending the active ingredient in a vegetable oil, such as peanut oil, olive oil, sesame oil or coconut oil, or in a mineral oil, such as liquid paraffin. Oily suspensions can contain a thickening agent, such as beeswax, hard paraffin or cetyl alcohol. Sweetening and flavoring agents, such as those described above, can be added to provide a palatable oral preparation. These compositions can be preserved by the addition of an antioxidant, such as ascorbic acid.

[0083] Dispersible powders or granules suitable for preparing aqueous suspension by adding water are provided in the mixture of active ingredient mixed with dispersing or wetting agent, suspending agent and one or more preservatives.Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above.Additional excipients, such as sweeteners, flavoring agents and coloring agents, may also be present.

[0084] The pharmaceutical composition of the present invention may also be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil, such as olive oil or peanut oil, a mineral oil, such as liquid paraffin, or a mixture thereof. Suitable emulsifiers may be natural gums, such as gum acacia or gum tragacanth, natural phosphatides, such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate and the condensation products of said partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners and flavoring agents.

[0085] Syrups and elixirs can be formulated with sweetening agents, such as glycerol, propylene glycol, sorbitol or sucrose. Such formulations can also contain demulcents, preservatives, flavoring agents and coloring agents. Oral liquid preparations can be prepared in combination with, for example, cyclodextrin, PEG and surfactants.

[0086] The pharmaceutical composition may be in the form of a sterile injectable aqueous or oily suspension. This suspension may be formulated by known techniques using suitable dispersing or wetting agents and suspending agents as mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, a solution in 1,3-butanediol. As acceptable vehicles and solvents that may be used, water, Ringer's solution and isotonic sodium chloride solution may be used. In addition, sterile, fixed oils are conventionally used as a solvent or suspending medium. For this purpose, any sterile, fixed oil may be used, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

[0087] The compounds of the present invention can also be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature, and therefore melts in the rectum to release the drug. Such materials include cocoa butter and polyethylene glycol. In addition, the compounds can be administered by intraocular delivery by using solutions or ointments. In addition, the compounds of interest can be delivered transdermally by using iontophoretic patches and the like. For topical use, creams, ointments, jellies, solutions or suspensions containing the compounds of the present invention are used. As used herein, topical application is also intended to include the use of mouthwashes and gargles.

[0088] The compounds of the present invention can also be coupled to carriers that are suitable polymers as targetable drug carriers. Such polymers include polyvinylpyrrolidone, pyran copolymers, polyhydroxy-propyl-methacrylamide-phenol, polyhydroxyethyl-aspartamide-phenol, or polyethylene oxide-polylysine substituted with palmitoyl residues. In addition, the compounds of the present invention can be coupled to carriers that are a class of biodegradable polymers useful for achieving controlled release of drugs, such as polylactic acid, polyglycolic acid, copolymers of polylactic acid and polyglycolic acid, polyepsilon caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates, and crosslinked or amphiphilic block copolymers of hydrogels. Polymers and semipermeable polymer matrices can be formed into shaped articles such as valves, stents, tubes, prostheses, etc.

[0089] C.How to use Without being bound to any particular theory, it is believed that the compounds and compositions of the present invention provide a therapeutic effect by inhibiting the binding of SDF-1 and / or I-TAC to the CXCR7 receptor. Thus, the compounds and compositions of the present invention can be used to treat or prevent diseases or disorders in mammals in which inhibition of the binding of SDF-1 and / or I-TAC to the CXCR7 receptor provides a therapeutic effect.

[0090] In one embodiment, a preferred method of inhibiting binding of the chemokines SDF-1 and / or I-TAC to the CXCR7 receptor comprises contacting a cell expressing the CXCR7 receptor with one or more of the aforementioned compounds for a period of time sufficient to inhibit binding of these chemokines to the CXCR7 receptor.

[0091] In some embodiments, the compounds and compositions of the invention are administered to a subject with cancer. In some cases, the CXCR7 modulators are used to treat cancer, e.g., carcinoma, glioma, mesothelioma, melanoma, lymphoma, leukemia (including acute lymphocytic leukemia), adenocarcinoma, breast cancer, ovarian cancer, cervical cancer, glioblastoma, leukemia, lymphoma, prostate cancer and Burkitt's lymphoma, head and neck cancer, colon cancer, colorectal cancer, non-small cell lung cancer, small cell lung cancer, esophageal cancer, gastric cancer, pancreatic cancer, hepatobiliary cancer, gallbladder cancer, small intestine cancer, rectal cancer, kidney cancer, renal cancer, bladder cancer, prostate cancer, penile cancer, urethral cancer, testicular cancer, cervical cancer, vaginal cancer, uterine cancer, ovarian cancer, thyroid cancer, parathyroid cancer, adrenal cancer, pancreatic endocrine cancer, carcinoid cancer, osteosarcoma, skin cancer, retinoblastoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma (for additional cancers, see Cancer: Principles and Practice (DeVita, VT et al. eds 1997); and brain and nerve dysfunction such as Alzheimer's disease, multiple sclerosis and demyelinating diseases; hypertensive disorders such as pulmonary arterial hypertension; kidney dysfunction; renal dysfunction; rheumatoid arthritis; allograft rejection; atherosclerosis (and elevated cholesterol levels); asthma; glomerulonephritis; contact dermatitis; inflammatory bowel disease; colitis; psoriasis; reperfusion injury; and other disorders and diseases described herein. In some embodiments, the subject does not have Kaposi's sarcoma, multicentric Castleman's disease, or AIDS-related primary effusion lymphoma.

[0092] The present invention also encompasses the administration of the compounds and compositions of the present invention to reduce angiogenesis in any subject in need thereof. For example, contacting CXCR7 with the compounds of the present invention to reduce CXCR7 activity and thereby reduce angiogenesis is useful for inhibiting the formation, growth and / or metastasis of tumors, particularly solid tumors. Descriptions of embodiments relating to regulated CXCR7 and angiogenesis are described, for example, in US Patent Application Publication No. 11 / 050,345.

[0093] Other disorders involving unwanted or problematic angiogenesis include rheumatoid arthritis; psoriasis; angiogenic diseases of the eye, such as diabetic retinopathy, retinopathy of prematurity, macular degeneration, corneal graft rejection, neovascular glaucoma, retrolental fibroplasia, rubeosis; Osler-Weber syndrome; myocardial angiogenesis; plaque angiogenesis; telangiectasia; hemophilic joints; angiofibromas; diseases of excessive or abnormal stimulation of endothelial cells, such as intestinal adhesions, Crohn's disease, skin diseases such as psoriasis, eczema and scleroderma, diabetes, diabetic retinopathy, retinopathy of prematurity, age-related macular degeneration, atherosclerosis, scleroderma, wound granulation and hypertrophic scars, i.e., keloids; diseases with angiogenesis as a pathological consequence, such as cat scratch disease and ulcers (Helicobacter pylori), may also be treated with the antibodies of the invention. Angiogenesis inhibitors can be used to prevent or inhibit adhesions, especially intraperitoneal or pelvic adhesions such as those resulting after laparotomy or laparoscopic surgery, and burn contractions.Other conditions that may be advantageously treated with angiogenesis inhibitors include transplant surgery, liver cirrhosis, pulmonary fibrosis after acute respiratory distress syndrome or other pulmonary fibrosis in newborns, temporary prosthetic implants, and prevention of scarring following postoperative adhesions between the brain and dura.Endometriosis, polyps, cardiac hypertrophy, and obesity can also be treated by inhibiting angiogenesis.These disorders may be accompanied by an increase in size or proliferation of other types of normal tissues, such as uterine fibroids, prostatic hypertrophy, and amyloidosis.The compounds and compositions of the present invention can be used prophylactically or therapeutically for any of the disorders or diseases described herein.

[0094] The reduction of CXCR7 activity using the compounds and compositions of the present invention can also be used to prevent neovascularization to effectively treat a host of disorders. Thus, for example, the reduction of angiogenesis can be used as part of the treatment for vascular disorders (e.g., hemangiomas and capillary proliferation in atherosclerotic plaques), muscle disorders (e.g., myocardial angiogenesis, myocardial infarction, or angiogenesis in smooth muscle), joints (e.g., arthritis, hemophilic joints, etc.), and other disorders involving angiogenesis. The promotion of angiogenesis can also be useful in the treatment of various physiological processes and diseases that require increased angiogenesis, such as the healing of wounds, fractures and burns, inflammatory diseases, ischemic heart, and peripheral vascular diseases. The compounds of the present invention can also provide benefits in conditions where normal blood flow is restricted, such as pulmonary hypertension.

[0095] The compounds and compositions of the present invention can also be used to promote wound healing. Although the present invention is not limited to a particular mechanism of action, antagonism of CXCR7 may instead allow endogenous ligands to bind to low affinity receptors, thereby inducing enhanced wound healing. For example, SDF-1 binds to both CXCR7 and CXCR4, but binds to CXCR4 with low affinity. Similarly, I-TAC binds to CXCR3 with lower affinity than I-TAC binds to CXCR7. By preventing the binding of these ligands to CXCR7, CXCR7 antagonists can allow the ligands to bind to other receptors, thereby promoting wound healing. Thus, antagonism of CXCR7 to promote wound healing may be mediated by a different mechanism than promoting wound healing by stimulating CXCR7 activity with agonists.

[0096] In addition to treating disorders and conditions associated with neovascularization, inhibition of angiogenesis can be used to regulate or prevent the occurrence of normal physiological conditions associated with neovascularization.Thus, for example, the compounds and compositions can be used as birth control.According to the present invention, reducing CXCR7 activity in the ovary or endometrium can attenuate the neovascularization associated with ovulation, embryo implantation, placentation, etc.

[0097] Inhibitors of angiogenesis also have other therapeutic uses. For example, the compounds and compositions of the present invention can be used to: (a) Panniculectomy and treatment of obesity. See, e.g., Kolonin et al., Nature Medicine 10(6):625-632 (2004). (b) Treatment of preeclampsia. See, e.g., Levine et al., N. Engl. J. Med. 350(7):672-683 (2004); Maynard, et al., J. Clin. Invest. 111(5):649-658 (2003). (c) Treatment of cardiovascular disease. See, e.g., March, et al., Am. J. Physiol. Heart Circ. Physiol. 287:H458-H463 (2004); Rehman et al., Circulation 109:1292-1298 (2004). (d) Treatment / prevention of ischemia-reperfusion injury by inhibiting the hypoxic prerequisites of mesenchymal stem cells and rescuing ischemic tissues, including but not limited to myocardium and transplanted kidney tissue. See, e.g., Liu, H., et al., PLoS One (2012) 7: e34608 and Zhang, S. et al., Biomed Pharmacother (2020) 127: 110168. (e) Treatment of Shiga toxin-associated hemolytic uremic syndrome. See, e.g., Petruzziello-Pellegrini, TNet al., J Clin Invest (2012) 122:759-776. (f) Treatment of pain, anxiety, stress, opioid addiction and other CNS conditions. See, e.g., Meyrath M. et al., Nat Commun (2020) 19:3033 and Zhu, Y. et al. (g) Treatment / prevention of acute renal failure by blocking deleterious migration of renal progenitor cells in the glomuleri. See, e.g., Mazzinghi, B, et al., J Exp Med (2008) 205:479-490 and Romaol, S. et al., Kidney Int (2018) 94:1111-1126. (h) Treatment of neuroinflammatory disorders, including but not limited to multiple sclerosis (MS). See, e.g., Williams, J Let al., Glia (2020) 68: 1361-1374.

[0098] Cancer Treatment More specifically, the present invention also provides a method for treating cancer. A preferred method for treating cancer comprises administering to a cancer patient a therapeutically effective amount of one or more of the above-mentioned compounds (or salts thereof) for a period of time sufficient to treat the cancer.

[0099] For therapeutic purposes, the compositions of the present invention may be administered orally, parenterally (e.g., intramuscular, intraperitoneal, intravenous, ICV, intracisternal injection or infusion, subcutaneous injection or implant), by inhalation spray, nasal, vaginal, rectal, sublingual or topical routes of administration, and may be formulated, alone or together, into suitable dosage unit formulations containing conventional non-toxic pharmacologic acceptable carriers, adjuvants and vehicles appropriate for each route of administration.

[0100] In some embodiments, the CXCR7 modulators of the present invention may be administered in combination with other suitable therapeutic agents, such as chemotherapeutic agents, radiation, and the like. It should be understood that such administration may occur before, after, or simultaneously with the second therapeutic agent, such that the therapeutic effect of the second therapeutic agent is enhanced compared to administering the second agent in the absence of the CXCR7 modulator. The selection of suitable agents for use in combination therapy may be performed by one of skill in the art according to conventional pharmaceutical principles. The combination of therapeutic agents may act synergistically to achieve treatment or prevention of various disorders, such as cancer, wounds, renal dysfunction, brain dysfunction, or neurological dysfunction. Using this approach, it may be possible to achieve therapeutic effects with lower dosages of each agent, thus reducing the possibility of adverse side effects.

[0101] In addition to primates, such as humans, various other mammals can be treated according to the method of the present invention. For example, mammals can be treated, including but not limited to, cows, sheep, goats, horses, dogs, cats, guinea pigs, rats, or other bovine, ovine, equine, canine, feline, rodent, or murine species. However, the method can also be practiced in other species, such as birds (e.g., chickens).

[0102] In the treatment or prevention of conditions requiring chemokine receptor modulation, suitable dosage levels are generally about 0.001-100 mg / kg of patient body weight per day, which may be administered in single or multiple doses. Preferably, dosage levels are about 0.01-25 mg / kg per day, more preferably about 0.05-10 mg / kg / day. Suitable dosage levels may be about 0.01-25 mg / kg per day, about 0.05-10 mg / kg per day, or about 0.1-5 mg / kg per day. Within this range, dosages may be 0.005-0.05, 0.05-0.5, or 0.5-5.0 mg / kg per day. For oral administration, the composition is preferably provided in the form of a tablet containing 1.0 to 1000 milligrams of active ingredient, particularly 1.0, 5.0, 10.0, 15.0, 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0, 400.0, 500.0, 600.0, 750.0, 800.0, 900.0 and 1000.0 milligrams of active ingredient to tailor the dosage to the patient being treated. The compound may be administered on a regimen of 1 to 4 times per day, preferably once or twice per day.

[0103] It will be understood, however, that the specific dosage level and frequency of administration for any particular patient may vary, and will depend on a variety of factors, including the activity of the specific compound used, the metabolic stability and length of action of that compound, the age, weight, genetic characteristics, general health, sex and diet of the subject, as well as the mode and time of administration, rate of excretion, drug combination, and the severity of the particular condition of the subject being treated.

[0104] The compounds and compositions of the present invention may be combined with other compounds and compositions that have utility in relation to the prevention and treatment of cancers and diseases or conditions associated with CXCR7 signaling. Such other drugs may be administered simultaneously or sequentially with the compounds or compositions of the present invention, by routes and amounts commonly used therefor. When the compounds or compositions of the present invention are used simultaneously with one or more other drugs, pharmaceutical compositions containing such other drugs in addition to the compounds or compositions of the present invention are preferred. Thus, pharmaceutical compositions of the present invention include those that also contain one or more other active ingredients or therapeutic agents in addition to the compounds or compositions of the present invention. Examples of other therapeutic agents that may be combined with the compounds or compositions of the present invention, administered separately, or in the same pharmaceutical composition include, but are not limited to, cisplatin, paclitaxel, methotrexate, cyclophosphamide, ifosfamide, chlorambucil, carmustine, carboplatin, vincristine, vinblastine, thiotepa, lomustine, semustine, 5-fluorouracil, and cytarabine. The weight ratio of the compounds of the present invention to the second active ingredient may vary and will vary depending on the effective dose of each ingredient. Generally, each effective dose is used.Therefore, for example, when the compound of the present invention is combined with a second anticancer drug, the weight ratio of the compound of the present invention to the second drug is generally within the range of about 1000:1 to about 1:1000, preferably about 200:1 to about 1:200.The combination of the compound of the present invention with other active ingredients is also generally within the above range, but in each case, the effective dose of each active ingredient is used.

[0105] Treatment of inflammation Additionally, the compounds and compositions of the present invention are useful for treating inflammation and can be combined with other compounds and compositions having therapeutic utility that may require treatment before, after, or simultaneously with the treatment of cancer or inflammation with the present compounds. Thus, combination methods and compositions are also components of the present invention for preventing and treating conditions or diseases of interest, such as inflammatory bowel disease, rheumatoid arthritis, osteoarthritis, psoriatic arthritis, polyarticular arthritis, multiple sclerosis, allergic diseases, psoriasis, atopic dermatitis, and asthma, as well as inflammatory or autoimmune disorders, conditions, and diseases including the above conditions.

[0106] For example, in the treatment or prevention of inflammation or autoimmunity or bone loss associated with, for example, arthritis, the compounds and compositions may be used in combination with compounds such as anti-inflammatory or analgesic agents, such as opiate agonists, lipoxygenase inhibitors, such as inhibitors of 5-lipoxygenase, cyclooxygenase inhibitors, such as cyclooxygenase-2 inhibitors, interleukin inhibitors, such as interleukin-1 inhibitors, NMDA antagonists, inhibitors of nitric oxide or inhibitors of nitric oxide synthesis, nonsteroidal anti-inflammatory agents or cytokine suppressive anti-inflammatory agents, such as acetaminophen, aspirin, codeine, fentanyl, ibuprofen, indomethacin, ketorolac, morphine, naproxen, phenacetin, piroxicam, steroidal analgesics, sufentanil, sulindac, tenidap, and the like. Similarly, the compounds and compositions may be administered with analgesics as listed above; enhancers such as caffeine, H2 antagonists (e.g., ranitidine), simethicone, aluminum hydroxide or magnesium hydroxide; decongestants such as phenylephrine, phenylpropanolamine, pseudoephedrine, oxymetazoline, epinephrine, naphazoline, xylometazoline, propylhexedrine or levodesoxyephedrine; cough suppressants such as codeine, hydrocodone, caramiphen, carbetapentane or dextromethorphan; diuretics; and sedating or non-sedating antihistamines.

[0107] As mentioned above, the compounds and compositions of the present invention can be used in combination with other drugs that are used to treat, prevent, suppress or improve the disease or condition for which the compounds and compositions of the present invention are useful. Such other drugs can be administered simultaneously or sequentially with the compounds or compositions of the present invention, by a route and in an amount generally used therefor. When the compounds or compositions of the present invention are used simultaneously with one or more other drugs, pharmaceutical compositions that contain such other drugs in addition to the compounds or compositions of the present invention are preferred. Thus, pharmaceutical compositions of the present invention include those that contain one or more other active ingredients or therapeutic agents in addition to the compounds or compositions of the present invention. Examples of other therapeutic agents that may be combined with the compounds or compositions of the invention, administered separately or in the same pharmaceutical composition, include (a) VLA-4 antagonists, (b) corticosteroids such as beclomethasone, methylprednisolone, betamethasone, prednisone, prenisolone, dexamethasone, fluticasone, hydrocortisone, budesonide, triamcinolone, salmeterol, salmeterol, salbutamol, formoterol; (c) cyclosporines (cyclosporine A, Sandimmune®, Neoral®), tacrolimus (FK-506, Prograf®), rapamycin (sirolimus), and the like. , Rapamune®), other FK-506 type immunosuppressants and mycophenolates, e.g. mycophenolate mofetil (CellCept®); (d) antihistamines (H1-histamine antagonists) such as brompheniramine, chlorpheniramine, dexchlorpheniramine, triprolidine, clemastine, diphenhydramine, diphenylpyraline, tripelennamine, hydroxyzine, methdilazine, promethazine, trimeprazine, azatadine, cyproheptadine, antazoline, pheniramine, pyrilamine, astemizole, terfenadine, loratadine, cetirizine, fexofenadine, descarboethoxyloratadine;(e) nonsteroidal antiasthmatics (e.g., terbutaline, metaproterenol, fenoterol, isoetharine, albuterol, bitolterol, and pirbuterol), theophylline, cromolyn sodium, atropine, ipratropium bromide, leukotriene antagonists (e.g., zafirlukast, montelukast, pranlukast, iralukast, povilkast, and SKB-106, 203), leukotriene biosynthesis inhibitors (zileuton, BAY-1005); (f) prolactins, propionic acid derivatives (e.g., aluminoprofen, benoxaprofen, bucloxic acid, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen, indoprofen, ketoprofen, miloprofen, naproxen, oxaprozin, pirprofen, pranoprofen, suprofen, tiaprofenic acid and tioxaprofen), acetic acid derivatives (e.g., indomethacin, acemetacin, alclofenac, clidanac, diclofenac, fenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac, isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin and zomepirac), fenamic acid derivatives (e.g., flufenamic acid, meclofenamic acid, mefenamic acid, niflumic acid and tolfenamic acid), biphenylcarboxylic acid derivatives (e.g., diflunisal and flufenisal), oxicams (e.g., isoxicam, piroxicam, sud ... (g) cyclooxygenase-2 (COX-2) inhibitors such as celecoxib (Celebrex®) and rofecoxib (Vioxx®); (h) inhibitors of phosphodiesterase type IV (PDE IV); (i) gold compounds such as auranofin and aurothioglucose; (j) etanercept (Enbrel®);(k) antibody therapeutics such as Orthoclone (OKT3), daclizumab (Zenapax®), basiliximab (Simulect®) and infliximab (Remicade®); (l) other antagonists of chemokine receptors, particularly CCR5, CXCR2, CXCR3, CCR2, CCR3, CCR4, CCR7, CX3CR1 and CXCR6; (m) lubricants or emollients such as petrolatum and lanolin; (n) keratolytic agents (e.g. tazarotene), (o) vitamins (p) PUVA; (q) anthralin (Drithrocreme®), (r) etretinate (Tegison®) and isotretinoin; (s) interferon beta-1β (Betaseron®), interferon beta-1α (Avonex®), azathioprine (Imurek®, Imuran®), glatiramer acetate (Cap (t) DMARDS such as methotrexate; (u) other compounds such as antimetabolites such as 5-aminosalicylic acid and its prodrugs, hydroxychloroquine, D-penicillamine, azathioprine, 6-mercaptopurine and methotrexate, DNA synthesis inhibitors such as hydroxyurea, and microtubule disrupting agents such as colchicine. The weight ratio of the compound of the present invention to the active ingredient of the present invention may vary and will vary depending on the effective dose of each ingredient. Generally, an effective dose of each will be used. Thus, for example, when the compound of the present invention is combined with an NSAID, the weight ratio of the compound of the present invention to the NSAID will generally range from about 1000:1 to about 1:1000, preferably from about 200:1 to about 1:200. Combinations of the compound of the present invention with other active ingredients will also generally be within the above range, but in each case an effective dose of each active ingredient will be used.

[0108] Methods for inducing progenitor / stem cell mobilization Still further, the compounds and compositions of the present invention may be useful for the treatment or amelioration of disorders or conditions in which progenitor / stem cell mobilization and thus progenitor / stem cell mobilization are effective or desirable, optionally using the compounds of the present invention, according to the procedures and protocols described in WO 05 / 000333, which is incorporated herein by reference in its entirety for all purposes. Conditions that may be improved or otherwise benefited include, for example, hematopoietic disorders, such as aplastic anemia, leukemia, drug-induced anemia, and hematopoietic disorders due to chemotherapy or radiotherapy. Still further, the compounds and compositions of the present invention may be used to provide enhanced transplant success during and following immunosuppressive treatment, as well as more effective wound healing and treatment of bacterial infections. Still further, the compounds and compositions of the present invention may be useful for the treatment or amelioration of disorders or conditions in which progenitor / stem cell mobilization and thus progenitor / stem cell mobilization are effective or desirable, optionally using the compounds of the present invention, according to the procedures and protocols described in WO 05 / 000333, which is incorporated herein by reference in its entirety for all purposes. Conditions that may be improved or otherwise benefited include, for example, hematopoietic disorders, such as aplastic anemia, leukemia, drug-induced anemia, and hematopoietic disorders due to chemotherapy or radiotherapy.Furthermore, the compounds and compositions of the present invention may be used during and following immunosuppressive treatment to enhance transplant success and provide more effective wound healing and treatment of bacterial infections.Optionally, after administration of the compounds of the present invention and mobilization of progenitor / stem cells, blood containing the mobilized cells is collected, and the mobilized cells are optionally purified, optionally expanded, and reintroduced into the same individual or a second individual (e.g., a matched donor) as needed.

[0109] Many different types of cells can be mobilized as needed. In some embodiments, hematopoietic progenitor cells (HSCs) are mobilized after administration of a compound or composition of the invention, and are optionally harvested and purified from other blood components. Optionally, mobilization of HSCs is induced by administering at least one compound of the invention together with one or more of granulocyte colony stimulating factor (G-CSF) or AMD3100 (1,1'-[1,4-phenylenebis(methylene)]bis[1,4,8,11-tetraazacyclotetradecane]octohydrobromide dihydrate) or a salt, racemate, or isomer thereof.

[0110] In some embodiments, endothelial progenitor cells (EPCs) are mobilized following administration of a compound or composition of the invention, and are optionally harvested and purified from other blood components. Optionally, mobilization of EPCs is induced by administering at least one compound of the invention together with one or more of vascular endothelial growth factor (VEGF), a VEGF agonist (including but not limited to a VEGF agonist antibody), or AMD3100, or a salt, racemate, or isomer thereof.

[0111] In some embodiments, mesenchymal stem cells (MSCs) or stromal progenitor cells (SPCs) are mobilized following administration of a compound or composition of the invention, and optionally harvested and purified from other blood components. Optionally, such mobilization is induced by administering at least one compound of the invention together with one or more of G-CSF, VEGF, a VEGF agonist (including but not limited to a VEGF agonist antibody), AMD3100, or a salt, racemate, or isomer thereof.

[0112] To immobilize progenitor or stem cells, suitable dosage levels are generally about 0.001-100 mg per kg of patient body weight per day, which can be administered in single or multiple doses. The compounds can be administered as a single dose, as doses over time, such as by iv or transdermal administration, or as multiple doses. The compounds of the invention can also be used in ex vivo treatment protocols to prepare cell cultures that are subsequently used to replenish the subject's blood cells. Ex vivo treatment can be performed on autologous cells harvested from peripheral blood or bone marrow from a matched donor or from an allograft.

[0113] The compounds of the present invention may be combined with other compounds and compositions that induce activation, proliferation or mobilization of progenitor / stem cells. In addition to the above, these include, but are not limited to, Fms-related tyrosine kinase 3 ligand (Flt3 ligand), interleukin 3 (IL-3), interleukin 7 (IL-7), interleukin 20 (IL-20), steel factor (SF) and granulocyte macrophage colony-stimulating factor (GM-CSF), which may provide therapeutic benefits before, after or at the same time as mobilization of progenitor / stem cells that require or may benefit from treatment. Thus, combination methods and compositions are also components of the present invention for preventing and treating conditions or diseases of interest. Additionally, the compounds of the present invention may provide benefits in conditions in which dysregulation of stem cell mobilization may play a role, such as heart disease and pulmonary hypertension.

[0114] Methods for diagnosing CXCR7-associated diseases and disorders Still further, the compounds and compositions of the present invention are useful for the diagnosis of diseases and disorders associated with CXCR7. Specifically, the compounds of the present invention can be prepared in a labeled form (e.g., radiolabeled) and used, for example, for the diagnosis of cancer. The labeled compounds of the present invention that bind to CXCR7 (e.g., antagonists or agonists) can be used to determine the level of CXCR7 in a mammalian subject. In some embodiments, a CXCR7 modulator is administered to a subject with cancer. In some cases, the labeled compound is directed to a cancer, e.g., carcinoma, glioma, mesothelioma, melanoma, lymphoma, leukemia, adenocarcinoma, breast cancer, ovarian cancer, cervical cancer, glioblastoma, leukemia, lymphoma, prostate cancer and Burkitt's lymphoma, head and neck cancer, colon cancer, colorectal cancer, non-small cell lung cancer, small cell lung cancer, esophageal cancer, gastric cancer, pancreatic cancer, hepatobiliary cancer, gallbladder cancer, small intestine cancer, rectal cancer, kidney cancer, bladder cancer, prostate cancer, penis cancer, urethral cancer, testicular cancer, cervical cancer, vaginal cancer, uterine cancer, ovarian cancer, thyroid cancer, parathyroid cancer, adrenal cancer, pancreatic endocrine cancer, carcinoid cancer, osteosarcoma, skin cancer, retinoblastoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma (for additional cancers, see Cancer: Principles and Practice (DeVita, VT et al. eds 1997) and to detect the onset of brain and nerve dysfunction, such as Alzheimer's disease and multiple sclerosis; renal dysfunction; rheumatoid arthritis; cardiac allograft rejection; atherosclerosis (and elevated cholesterol levels); asthma; glomerulonephritis; contact dermatitis; inflammatory bowel disease; colitis; psoriasis; reperfusion injury; and other disorders and diseases described herein. In some embodiments, the subject does not have Kaposi's sarcoma, multicentric Castleman's disease, or AIDS-related primary effusion lymphoma. Because CXCR7 is often expressed in cancer cells, but not in non-cancerous cells, it is typically desirable to administer an antagonist of CXCR7 to a subject at risk of having cancer.

[0115] Various imaging and detection methods can be used to detect cancer. In some embodiments, direct methods such as magnetic resonance imaging ("MRI"), positron emission tomography ("PET") and single photon emission computed tomography ("SPECT") can be used to assess CXCR7 biodistribution in the body. Each of these methods can detect the distribution of appropriately labeled compounds (typically bound to CXCR7) in the body, if the compound contains atoms with appropriate nuclear properties. MRI detects paramagnetic atomic nuclei; PET and SPECT detect the emission of particles resulting from the decay of radioactive nuclei.

[0116] For procedures involving PET, it is necessary to incorporate a suitable positron-emitting radionuclide. There are a relatively small number of positron-emitting isotopes that are suitable for labeling therapeutic agents. Carbon isotopes 11 C has been used in PET, but has a short half-life of 20.5 minutes. Therefore, facilities for synthesis and use typically use precursor 11 C is another useful isotope close to the cyclotron where the starting material is produced. 18 F has a half-life of 110 minutes, allowing sufficient time for incorporation into a radiolabeled tracer, purification, and administration into a human or animal subject. Other isotopes have even shorter half-lives. 13 N has a half-life of 10 minutes, 15 O has a shorter half-life of 2 minutes. However, both emissions 11 C isotopes, and PET studies have been performed with these isotopes (see Clinical Positron Emission Tomography, Mosby Year Book, 1992, KF Hubner, et al., Chapter 2).

[0117] SPECT imaging uses isotopic tracers that are gamma-emitters. Although the range of useful isotopes is greater than PET, the three-dimensional resolution of SPECT imaging is low. However, in some cases, SPECT is used to obtain clinically significant information regarding compound binding, localization, and clearance rates. One useful isotope for SPECT imaging is γ-emitter, γ-emitter, with a half-life of 13.3 hours. 123 I. 123 Compounds labeled with I can be transported up to about 1000 miles from the manufacturing site, or the isotope itself can be transported for on-site synthesis. 85% of the isotope's emission is 159 KeV photons, which is easily measured by currently used SPECT instruments. Other halogen isotopes may also be useful for PET or SPECT imaging or conventional tracer labeling. These include, as having usable half-lives and emission characteristics: 75 Br, 76 Br, 77 Br and 82 Br is one example.

[0118] In view of the above, the present invention provides a method for imaging a tumor, organ or tissue, comprising: (a) administering to a subject in need of such imaging a compound of formula I in a radiolabeled or detectable form; (b) detecting the compound and determining where in the subject the compound is concentrated; The present invention provides a method comprising:

[0119] The present invention further provides a method for detecting elevated levels of CXCR7 in a sample, comprising the steps of: (a) contacting a sample suspected of having an elevated level of CXCR7 with a radiolabeled or detectable form of a compound of formula I; (b) determining the level of a compound bound to CXCR7 present in the sample to determine the level of CXCR7 present in the sample; (c) comparing the level determined in step (b) to a control sample to determine whether an elevated level of CXCR7 is present in the sample; The present invention provides a method comprising:

[0120] As with the therapeutic methods described herein, administration of the labeled compound may be by any of the routes commonly used to introduce a compound into ultimate contact with the tissue to be evaluated, which are well known to those skilled in the art. Although multiple routes may be used to administer a particular composition, a particular route may often provide a more rapid and effective diagnosis than another route.

[0121] Combination therapy The inhibitor of CXCR7 can be provided alone or in combination with one or more other drugs. Possible combination patterns can include, for example, additional antiangiogenic factors and / or chemotherapeutic agents (e.g., cytotoxic agents) or radiation, cancer vaccines, immunomodulators, antivascular agents, signal transduction inhibitors, antiproliferative agents or apoptosis inducers. EXAMPLES

[0122] The following examples are offered to illustrate, but not to limit, the claimed invention.

[0123] Reagents and solvents used below can be obtained from commercial sources such as Aldrich Chemical Co. (Milwaukee, Wisconsin, USA). 1H-NMR spectra were recorded on a Varian Mercury 400MHz NMR spectrometer. Important peaks are given relative to TMS and are listed in the following order: multiplicity (s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet) and proton number. Mass spectrometry results are reported as the ratio of mass to charge followed by the relative abundance of each ion (in parentheses). In this example, a single m / e value is reported for the M+H (or MH, where noted) ion containing the most common atomic isotopes. The isotope pattern corresponds to the expected formula in all cases. Electrospray ionization (ESI) mass spectrometry was performed on a Hewlett-Packard MSD electrospray mass spectrometer using an HP1100 HPLC for sample delivery. Typically, the analytes were dissolved at 0.1 mg / mL in methanol and 1 microliter was injected into the mass spectrometer with the delivery solvent, scanning from 100 to 1500 daltons. All of the compounds could be analyzed in positive ESI mode using acetonitrile / water with 1% formic acid as the delivery solvent. The compounds provided below could also be analyzed in negative ESI mode using acetonitrile / water with 2 mM NHOAc as the delivery system.

[0124] The following abbreviations are used in the examples and throughout the description of the invention: rt, room temperature; HPLC, high pressure liquid chromatography; TFA, trifluoroacetic acid; LC-MSD, liquid chromatography / mass selective detector; LC-MS, liquid chromatography / mass spectrometer; Pd2dba3, tris(dibenzylideneacetone)dipalladium; THF, tetrahydrofuran; DMF, dimethylformamide or N,N-dimethylformamide; DCM, dichloromethane; DMSO, dimethylsulfoxide; TLC, thin layer chromatography; KHMDS, potassium hexamethyldisilazane; ES, electrospray; sat., saturated.

[0125] Compounds within the scope of the present invention can be synthesized using a variety of reactions known to those skilled in the art, as described below. Those skilled in the art will also recognize that alternative methods may be employed to synthesize the target compounds of the present invention, and that the approaches described in the main body of this specification, while not exhaustive, provide widely applicable and practical routes to compounds of interest.

[0126] Certain molecules claimed in this patent can exist in different enantiomeric and diastereomeric forms, and all such variants of these compounds are claimed.

[0127] The detailed description of the experimental procedures used to synthesize compounds of interest herein results in molecules that are illustrated by physical data that identifies the compounds and structural diagrams that relate to the compounds.

[0128] Those skilled in the art will also recognize that acids and bases are frequently used during standard workup procedures in organic chemistry. Salts of parent compounds may also be prepared during the laboratory procedures described within this patent, where they have the required inherent acidity or basicity.

[0129] Example 1: 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2-((2-(pyridin-2-yl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] Step a: A mixture of tert-butyl 3-oxoazetidine-1-carboxylate (15 g, 88 mmol), ethyl hydrogen malonate (17 g, 130 mmol) and ammonium formate (20 g, 260 mmol) in 120 mL of EtOH was stirred at 85° C. for 3 h. The volatiles were removed in vacuo and the residue was diluted with Et2O and washed with saturated NaHCO3(aq). The organic layer was dried over MgSO4, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography to give tert-butyl 3-amino-3-(2-ethoxy-2-oxoethyl)azetidine-1-carboxylate. MS: (ES) m / z C 12 H 23 N2O4[M+H] + Calculated value: 259.2, measured value: 259.3.

[0130] Step b: To a solution of tert-butyl 3-amino-3-(2-ethoxy-2-oxoethyl)azetidine-1-carboxylate (1.8 g, 7.1 mmol), 5-(2,4-difluorophenyl)isoxazole-3-carboxylic acid (1.6 g, 7.1 mmol) and DIPEA (1.3 mL, 7.5 mmol) in 40 mL of DCM was added HATU (2.8 g, 7.4 mmol). The mixture was stirred at room temperature for 2 h. The reaction was quenched with saturated NH4Cl(aq) and the phases were separated. The aqueous phase was extracted with DCM and the combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The crude material was purified by silica gel column chromatography to give tert-butyl 3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-3-(2-ethoxy-2-oxoethyl)azetidine-1-carboxylate. MS: (ES) m / z C 22 H 26 F2N3O6[M+H] + Calculated value: 466.2, actual value: 466.2.

[0131] Step c: A mixture of tert-butyl 3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-3-(2-ethoxy-2-oxoethyl)azetidine-1-carboxylate (1.2 g, 3.3 mmol) in 4:1 DCM / TFA (20 mL) was stirred for 1 h. The contents were concentrated under reduced pressure to give ethyl acetate 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)azetidin-3-yl). MS: (ES) m / z C 17 H 18 F2N3O4[M+H] + Calculated value: 366.1, actual value: 366.1.

[0132] step d: To a mixture of ethyl acetate 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)azetidin-3-yl) (800 mg, 2.0 mmol), triethylamine (0.55 mL, 4.0 mmol) and 4-hydroxy-4-methylcyclohexan-1-one (510 mg, 4.0 mmol) in 4:1 DCM / MeOH (20 mL) was added NaBH(OAc)3 (840 mg, 4.0 mmol). The contents were stirred at room temperature for 1 h. The reaction was quenched with saturated NaHCO3(aq) and extracted with DCM. The combined organic layers were dried over MgSO4, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography to give ethyl acetate 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl) (mixture of cis / trans isomers). MS: (ES) m / z C 24 H 30 F2N3O5[M+H] + The calculated value was 478.2, and the actual value was 478.5.

[0133] Step e: To a solution of ethyl acetate 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl) (600 mg, 1.2 mmol) in 1:1 THF / H2O (20 mL) was added NaOH (300 mg, 7.5 mmol). The mixture was stirred at room temperature overnight. The reaction was quenched with 1N HCl (10 mL) and MeCN. The contents were concentrated under reduced pressure. The residue was triturated with acetone and filtered through Celite. The filtrate was concentrated to give 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (mixture of cis / trans isomers). MS: (ES) m / z C 22 H 26 F2N3O5[M+H] + Calculated value: 450.2, actual value: 450.2.

[0134] Step f: To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (100 mg, 0.21 mmol) in DCM (2 mL) was added 2-(pyridin-2-yl)propan-2-amine dihydrochloride (43 mg, 0.21 mmol), DIPEA (0.14 mL, 0.82 mmol) and HATU (82 mg, 0.22 mmol). The mixture was stirred for 1 h and then concentrated under reduced pressure. The residue was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2-((2-(pyridin-2-yl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0135] Isomer 1: 1H NMR(400MHz,DMSO-d6)δ 8.59-8.49(m,1H),8.22-8.12(m,1H),8.06(d,J=8.1Hz,1H),7.83-7.77(m,1H),7.67-7.58(m,1H),7.25(dd,J=20.8,9.8Hz,2H),7.15 -7.07(m,1H),4.58-4.27(m,4H),3.20-3.11(m,3H),1.83-1.73(m,2H),1.56-1.44(m,9H),1.37-1.15(m,4H),1.21(s,3H).MS:(ES)m / z C 30 H 36 F2N5O4[M+H] + Calculated value: 568.3, actual value: 568.3.

[0136] Isomer 2: 1 H NMR (400MHz, CD3OD) δ 8.54-8.48(m,1H),8.06(m,2H),7.71(d,J=8.0Hz,1H),7.56-7.50(m,1H),7 .32-7.17(m,2H),7.11(dd,J=10.4,3.5Hz,1H),4.52(d,J=11.8Hz,1H),4.49 -4.46(m,2H),4.37(d,J=11.7Hz,1H),3.16-3.10(m,3H),1.80-1.66(m,4H), 1.66(s,6H),1.62-1.52(m,2H),1.46-1.39(m,2H),1.20(s,3H).MS:(ES)m / z C 30 H 36 F2N5O4[M+H] + Calculated value: 568.3, measured value: 568.4.

[0137] Example 2: 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2-((2-phenylpropan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a solution of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-phenylpropan-2-amine (31 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The reaction mixture was stirred at room temperature for 1 h and then quenched with 3 drops of HO. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2((2-phenylpropan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0138] Isomer 1: 1 H NMR(400MHz,DMSO-d6)δ 9.61-9.48(m,1H),8.41-8.36(s,1H),8.19-8.07(m,1H),7.67-7.58( m,1H),7.42-7.32(m,1H),7.31-7.18(m,3H),7.16-7.02(m,3H),4.49- 4.27(m,4H),3.31-3.16(m,1H),3.14-3.04(m,2H),1.88-1.73(m,2H),1.61-1.46(m,8H),1.38-1.15(m,4H),1.12-1.02(m,3H).MS:(ES)m / z C 31 H 37 F2N4O4[M+H] + Calculated value: 567.3, actual value: 567.3.

[0139] Isomer 2: 1H NMR(400MHz,DMSO-d6)δ 9.50(s,1H),8.36-8.32(m,1H),8.17-8.07(m,1H),7.68-7.59(m,1H),7.41-7.34(m,1H),7.31-7.17(m,3H),7.14- 7.02(m,3H),4.46-4.26(m,4H),3.19-3.01(m,3H),1.73-1.37(m,12H),1.30-1.16(m,2H),1.10(s,3H).MS:(ES)m / z C 31 H 37 F2N4O4[M+H] + Calculated value: 567.3, actual value: 567.3.

[0140] Example 3: 5-(2,4-difluorophenyl)-N-3-(2-((2-(3-fluorophenyl)propan-2-yl)amino)-2-oxoethyl)-(1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a solution of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-(3-fluororophenyl)propan-2-amine (32 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The reaction mixture was stirred at room temperature for 1 h and then quenched with 3 drops of HO. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-3-(2-((2-(3-fluorophenyl)propan-2-yl)amino)-2-oxoethyl)-(1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0141] Isomer 1: 1H NMR (400MHz, DMSO-d6) δ 9.61-9.48(m,1H),8.41-8.36(s,1H),8.16-8.02(m,1H),7.65-7.56(m,1H),7. 39-7.30(m,1H),7.27-7.09(m,2H),7.08-7.01(m,1H),7.01-6.93(m,1H),6.92 -6.82(m,1H),4.50-4.25(m,4H),3.22(s,1H),3.18-3.08(m,2H),1.78(d,J=8. 5Hz,2H),1.58-1.42(m,8H),1.38-1.15(m,4H),1.11-1.01(m,3H).MS:(ES)m / z C 31 H 36 F3N4O4[M+H] + Calculated value: 585.3, actual value: 585.5.

[0142] Isomer 2: 1 H NMR(400MHz,DMSO-d6)δ 9.50(s,1H),8.40-8.35(m,1H),8.14-8.04(m,1H),7.67-7.55(m,1H),7 .35(dd,J=8.8,8.8Hz,1H),7.28-7.08(m,2H),7.08-7.01(m,1H),7.00-6 .82(m,2H),4.48-4.19(m,4H),3.10-3.01(m,3H),1.72-1.52(m,4H),1.4 8(d,J=2.4Hz,8H),1.28-1.13(m,2H),1.08(d,J=2.5Hz,3H).MS:(ES)m / z C 31 H 36 F3N4O4[M+H] + Calculated value: 585.3, actual value: 585.5.

[0143] Example 4: N-(3-(2-((2-(3-chlorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide [ka] Step a: To a solution of 3-chlorobenzonitrile (1.00 g, 7.30 mmol) in ether (100 mL) was added a 3.0 M solution of methylmagnesium bromide (7.30 mL, 21.9 mmol) dropwise, followed by titanium(IV) isopropoxide (2.16 mL, 7.30 mmol). The mixture was refluxed overnight under N2, then cooled to 0 °C and quenched with water, followed by 10% NaOH(aq). The contents were filtered through a plug of Celite and washed with DCM. The organic layer of the filtrate was collected, dried over Na2SO4, concentrated under reduced pressure and purified by silica gel column chromatography to give 2-(3-chlorophenyl)propan-2-amine. MS: (ES) m / z C9H 13 ClN[M+H] + The calculated value was 170.1 and the measured value was 170.2.

[0144] Step b: To a solution of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-(3-chlorophenyl)propan-2-amine (34 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The reaction mixture was stirred at room temperature for 1 h and then quenched with 3 drops of HO. The mixture was purified by preparative HPLC to give the two separated isomers of N-(3-(2-((2-(3-chlorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide.

[0145] Isomer 1: 1H NMR(400MHz,CD3OD)δ 8.58(s,1H),8.47(s,1H),8.12-7.96(m,1H),7.38-7.05(m,7H),4.58-4.35(m,4H),3.30-3.09(m,3H),2.03-1. 91(m,2H),1.74-1.64(m,2H),1.63-1.55(m,6H),1.55-1.45(m,2H),1.44-1.25(m,2H),1.20(s,3H).MS:(ES)m / z C 31 H 36 ClF2N4O4[M+H] + The calculated value was 601.2, and the measured value was 601.3.

[0146] Isomer 2: 1 H NMR(400MHz,CD3OD)δ 8.49(s,1H),8.47(s,1H),8.10-8.02(m,1H),7.32-7.07(m,7H),4.56-4.33(m,4H),3.30-3. 07(m,3H),1.86-1.70(m,4H),1.65-1.49(m,8H),1.48-1.35(m,2H),1.20(s,3H).MS:(ES)m / z C 31 H 36 ClF2N4O4[M+H] + The calculated value was 601.2, and the measured value was 601.3.

[0147] Example 5: 5-(2,4-difluorophenyl)-N-3-(2-((2-(2,5-difluorophenyl)propan-2-yl)amino)-2-oxoethyl)-(1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] Step a: To a solution of 2,5-difluorobenzonitrile (1.00 g, 7.19 mmol) in ether (100 mL) was added a 3.0 M solution of methylmagnesium bromide (7.30 mL, 21.9 mmol) followed by dropwise addition of titanium(IV) isopropoxide (2.16 mL, 7.30 mmol). The resulting mixture was refluxed overnight under N2, then cooled to 0 °C and quenched with water followed by 10% NaOH(aq). The contents were filtered through a plug of Celite and washed with DCM. The organic layer of the filtrate was collected, dried over Na2SO4, concentrated under reduced pressure and purified by silica gel column chromatography to give 2-(2,5-difluorophenyl)propan-2-amine. MS: (ES) m / z C9H 12 F2N[M+H] + The calculated value was 172.1, and the measured value was 172.2.

[0148] Step b: To a solution of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-(2,5-difluorophenyl)propan-2-amine (35 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The reaction mixture was stirred at room temperature for 1 h and then quenched with 3 drops of HO. The mixture was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(3-(2-((2-(2,5-difluorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0149] Isomer 1: 1H NMR(400MHz,CD3OD)δ 8.65(s,1H),8.55(s,1H),8.14-8.02(m,1H),7.32-7.18(m,2H),7.15-7.02(m,2H),6.98-6.74(m,2H),4.57- 4.33(m,4H),3.30-3.07(m,3H),1.97(s,2H),1.75-1.56(m,8H),1.55-1.30(m,4H),1.21(s,3H).MS:(ES)m / z C 31 H 35 F4N4O4[M+H] + Calculated value: 603.3, measured value: 603.5.

[0150] Isomer 2: 1 H NMR(400MHz,CD3OD)δ 8.57(s,1H),8.55(s,1H),8.13-8.03(m,1H),7.33-7.18(m,2H),7.14-7.02(m,2H),6.96-6.87(m,1H),6.86-6.75(m,1H) ,4.55-4.30(m,4H),3.15-3.08(m,3H),1.85-1.71(m,4H),1.66-1.51(m,8H),1.47-1.35(m,2H),1.20(s,3H).MS:(ES)m / z C 31 H 35 F4N4O4[M+H] + Calculated value: 603.3, measured value: 603.5.

[0151] Example 6: 5-(2,4-difluorophenyl)-N-(3-(2-((2-(3,5-difluorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] Step a: To a solution of 3,5-difluorobenzonitrile (1.00 g, 7.19 mmol) in ether (100 mL) was added a 3.0 M solution of methylmagnesium bromide (7.30 mL, 21.9 mmol) followed by dropwise addition of titanium(IV) isopropoxide (2.16 mL, 7.30 mmol). The resulting mixture was refluxed overnight under N2, then cooled to 0 °C and quenched with water followed by 10% NaOH(aq). The mixture was filtered through a plug of Celite and washed with DCM. The organic layer of the filtrate was collected, dried over Na2SO4, concentrated under reduced pressure and purified by silica gel column chromatography to give 2-(3,5-difluorophenyl)propan-2-amine. MS: (ES) m / z C9H 12 F2N[M+H] + The calculated value was 172.1, and the measured value was 172.2.

[0152] Step b: To a solution of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-(3,5-difluorophenyl)propan-2-amine (35 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The reaction mixture was stirred at room temperature for 1 h and then quenched with 3 drops of HO. The crude material was purified by reverse phase HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(3-(2-((2-(3,5-difluorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0153] Isomer 1: 1H NMR(400MHz,CD3OD)δ 8.58(s,1H),8.49(s,1H),8.11-8.01(m,1H),7.32-7.18(m,2H),7.13-7.06(m,1H),6.95-6.84(m,2H),6.76-6.65(m,1H),4.58-4. 35(m,4H),3.30-3.09(m,3H),2.05-1.92(m,2H),1.75-1.65(m,2H),1.64-1.46(m,8H),1.45-1.31(m,2H),1.21(s,3H).MS:(ES)m / z C 31 H 35 F4N4O4[M+H] + Calculated value: 603.3, measured value: 603.5.

[0154] Isomer 2: 1 H NMR(400MHz,CD3OD)δ 8.51(s,1H),8.48(s,1H),8.11-8.01(m,1H),7.31-7.18(m,2H),7.10(dd,J=10.1,3.5Hz,1H),6.92-6.83(m,2H),6.74-6.66(m ,1H),4.55-4.34(m,4H),3.30-3.11(m,3H),1.86-1.70(m,4H),1.66-1.50(m,8H),1.48-1.34(m,2H),1.20(s,3H).MS:(ES)m / z C 31 H 35 F4N4O4[M+H] + Calculated value: 603.3, measured value: 603.5.

[0155] Example 7: N-(3-(2-((2-(3-cyanophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide [ka] Step a: To a solution of tert-butyl (2-(3-bromophenyl)propan-2-yl)carbamate (1.00 g, 3.18 mmol) in 40 mL of dioxane was added Zn(CN)2 (0.747 g, 6.36 mmol). The contents were sparged with N2 for 10 min, followed by the addition of Pd(dppf)Cl2 (0.233 g, 0.318 mmol) and the resulting mixture was heated under N2 for 4 h. The reaction was cooled to room temperature and quenched with saturated NH4Cl(aq), followed by extraction with DCM. The organic layer was collected, dried over MgSO4, concentrated under reduced pressure and purified by silica gel column chromatography to give tert-butyl (2-(3-cyanophenyl)propan-2-yl)carbamate. MS: (ES) m / z C 15 H 21 N2O2[M+H] + Calculated value: 261.2, actual value: 261.2.

[0156] Step b: To a solution of tert-butyl (2-(3-cyanophenyl)propan-2-yl)carbamate (0.50 g, 1.9 mmol) in 8 mL of DCM was added trifluoroacetic acid. The reaction mixture was stirred at room temperature overnight and then quenched with saturated NaHCO3(aq). The contents were extracted with DCM and the organic layer was collected, dried over MgSO4, and concentrated under reduced pressure to give 3-(2-aminopropan-2-yl)benzonitrile. MS: (ES) m / z C 10 H 13 N2[M+H] + The calculated value was 161.1, and the actual value was 161.2.

[0157] Step c: To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 3-(2-aminopropan-2-yl)benzonitrile (32 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The contents were stirred at room temperature for 1 h and then quenched with 3 drops of HO. The mixture was purified by preparative HPLC to give the two separated isomers of N-(3-(2-((2-(3-cyanophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide.

[0158] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.61(s,1H),8.52(s,1H),8.12-8.02(m,1H),7.71-7.59(m,2H),7.56-7.49(m,1H),7.46-7.35(m,1H),7.30-7.17(m,2H),7.14-7. 07(m,1H),4.61-4.32(m,4H),3.30-3.09(m,2H),2.05-1.91(m,2H),1.77-1.45(m,8H),1.45-1.27(m,1H),1.20(s,3H).MS:(ES)m / z C 32 H 36 F2N5O4[M+H] + Calculated value: 592.3, actual value: 592.3.

[0159] Isomer 2: 1H NMR(400MHz,CD3OD)δ 8.55(s,1H),8.52(s,1H),8.11-8.03(m,1H),7.68-7.59(m,2H),7.55-7.49(m,1H),7.45-7.35(m,1H),7.31-7.17(m,2H),7.13-7. 07(m,1H),4.56-4.33(m,4H),3.30-3.11(m,3H),1.86-1.70(m,4H),1.66-1.50(m,8H),1.48-1.33(m,2H),1.20(s,3H).MS:(ES)m / z C 32 H 36 F2N5O4[M+H] + Calculated value: 592.3, actual value: 592.3.

[0160] Example 8: 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-((2-(3-methoxyphenyl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] Step a: To a solution of 3-methoxybenzonitrile (5.0 g, 29.8 mmol) in ether (120 mL) was added a 3.0 M solution of methylmagnesium bromide (29.8 mL, 89.5 mmol) dropwise, followed by titanium(IV) isopropoxide (8.82 mL, 29.8 mmol). The mixture was refluxed overnight under N2, then cooled to 0 °C and quenched with water followed by 10% NaOH(aq). The contents were filtered through a plug of Celite and washed with DCM. The organic layer of the filtrate was collected, dried over Na2SO4, concentrated under reduced pressure and purified by silica gel column chromatography to give 2-(3-methoxyphenyl)propan-2-amine. MS: (ES) m / z C 10 H 16 NO[M+H] + The calculated value was 166.1, and the actual value was 166.0.

[0161] Step b: To a solution of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-(3-methoxyphenyl)propan-2-amine (40 mg, 0.24 mmol), HATU (100 mg, 0.26 mmol) and DIPEA (0.57 mL, 0.82 mmol). The reaction mixture was stirred at room temperature for 1 h and then quenched with 3 drops of HO. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-((2-(3-methoxyphenyl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0162] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.50(s,1H),8.41(s,1H),8.10-8.02(m,1H),7.30-7.18(m,2H),7.14-7.06(m,1H),6.92-6.72(m,1H),6.69(d,J=8.0Hz, 1H),4.57-4.35(m,4H),3.70-3.64(m,3H),3.18-3.08(m,2H),1.95(bs,2H),1.73-1.30(m,13H),1.20(s,3H).MS:(ES)m / z C 32 H 39 F2N4O5[M+H] + Calculated value: 597.3, actual value: 597.5.

[0163] Isomer 2: 1H NMR(400MHz,CD3OD)δ 8.41(s,1H),8.12-8.03(m,1H),7.30-7.18(m,2H),7.14-7.06(m,2H),6.87(d,J=8.0Hz,1H),6.83(s,1H),6.69( d,J=7.6Hz,1H),4.52-4.34(m,3H),3.65(s,3H),3.12-3.09(m,3H),1.90-1.30(m,15H),1.20(s,3H).MS:(ES)m / z C 32 H 39 F2N4O5[M+H] + Calculated value: 597.3, actual value: 597.5.

[0164] Example 9: 5-(2,4-difluorophenyl)-N-(3-(2-((2-(5-fluoro-2-methoxyphenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] Step a: To a solution of 5-fluoro-2-methoxybenzonitrile (1.00 g, 5.46 mmol) in ether (100 mL) was added a 3.0 M solution of methylmagnesium bromide (5.55 mL, 16.7 mmol) dropwise, followed by titanium(IV) isopropoxide (1.65 mL, 5.46 mmol). The resulting mixture was refluxed overnight under N2, then cooled to 0 °C and quenched with water, followed by 10% NaOH(aq). The contents were filtered through a plug of Celite and washed with DCM. The organic layer of the filtrate was collected, dried over Na2SO4, concentrated under reduced pressure and purified by silica gel column chromatography to give 2-(5-fluoro-2-methoxyphenyl)propan-2-amine. MS: (ES) m / z C 10 H 15 FNO[M+H] + The calculated value was 184.1, and the actual value was 184.0.

[0165] Step b: To a solution of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-(5-fluoro-2-methoxyphenyl)propan-2-amine (37 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The reaction mixture was stirred at room temperature for 1 h and then quenched with 3 drops of HO. The mixture was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-((2-(5-fluoro-2-methoxyphenyl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0166] Isomer 1: 1 H NMR(400MHz,DMSO-d6)δ 9.66-9.47(m,1H),8.23-8.14(m,1H),8.12-8.03(m,1H),7.65-7.56(m,1 H),7.38-7.30(m,1H),7.24-7.16(m,1H),6.93-6.78(m,3H),4.46-4.22( m,4H),3.61(s,3H),3.30-3.22(m,1H),3.09-2.98(m,2H),1.85-1.71(m, 2H),1.61-1.46(m,8H),1.37-1.14(m,4H),1.09-1.00(m,3H).MS:(ES)m / z C 32 H 38 F3N4O5[M+H] + Calculated value: 615.3, measured value: 615.3.

[0167] Isomer 2: 1H NMR(400MHz,DMSO-d6)δ 9.54(s,1H),8.19-8.14(m,1H),8.13-8.02(m,1H),7.65-7.56(m,1H),7.39-7.30(m,1H),7.26-7.15(m,1H),6.94-6.78(m ,3H),4.43-4.21(m,4H),3.61(s,3H),3.15-2.96(m,3H),1.71-1.34(m,12H),1.27-1.12(m,2H),1.07(s,3H).MS:(ES)m / z C 32 H 38 F3N4O5[M+H] + Calculated value: 615.3, measured value: 615.3.

[0168] Example 10: N-(1-cyclohexyl-3-(2-((2-(3-fluorophenyl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide [ka] Step a: To a solution of methyl 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)azetidin-3-yl) acetate (1.4 g, 3.6 mmol) in 27 mL of DCM was added cyclohexanone (0.62 mL, 6.0 mmol) and DIPEA (2.1 mL, 12.0 mmol). After 30 min, NaBH(OAc)3 (1.68 g, 8.0 mmol) was added and the mixture was stirred at room temperature. Upon completion, the reaction was quenched with H2O and the aqueous layer was extracted with EtOAc. The organic layers were combined, dried over sodium sulfate, filtered and concentrated. The resulting residue was purified by silica gel column chromatography to give methyl 2-(1-cyclohexyl-3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)azetidin-3-yl) acetate. MS: (ES) m / z C 22 H 26 F2N3O4[M+H] + Calculated value: 434.2, measured value: 434.2.

[0169] Step b: To a solution of methyl 2-(1-cyclohexyl-3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)azetidin-3-yl)acetate (1.55 g, 3.9 mmol) in 10.7 mL of THF was added a solution of 1 M LiOH (10.7 mL, 10.7 mmol). The reaction mixture was stirred at room temperature and then quenched with 1 N HCl. The volatiles were removed in vacuo and the resulting solid was filtered, washed and collected to give 2-(1-cyclohexyl-3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)azetidin-3-yl)acetic acid. MS: (ES) m / z C 21 H 24 F2N3O4[M+H] + The calculated value was 420.2, and the measured value was 420.1.

[0170] Step c: To a solution of 2-(1-cyclohexyl-3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)azetidin-3-yl)acetic acid (75 mg, 0.18 mmol) in 1.2 mL of DMF was added 2-(3-fluorophenyl)propan-2-amine (41 mg, 0.30 mmol), DIPEA (0.16 mL, 0.92 mmol) and HATU (0.20 g, 0.53 mmol). The reaction mixture was stirred at room temperature for 2 h and then quenched with HO. The aqueous layer was extracted with EtOAc and the organic layers were combined, dried over sodium sulfate, filtered and concentrated. The crude residue was purified by silica gel column chromatography followed by preparative HPLC to give N-(1-cyclohexyl-3-(2-((2-(3-fluorophenyl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide. 1H NMR(400MHz,DMSO-d6)δ:9.53(d,J=26.2Hz,1H),8.38(d,J=2.9Hz,1H),8.08(dddd,J=8.8,8.8,6.4,2.5Hz,1H),7.60(ddd,J=10.2,10.2,2.0Hz 1H),7.34(ddd,J=8.4,8.4,2.4Hz,1H),7.23(dd,J=20.4,3.1Hz,1H),7.16-7.06(m,1 H),7.03(d,J=8.0Hz,1H),6.95(dddd,J=11.0,2.2,2.2,2.2Hz,1H),6.86(dddd,J=8. 3,8.3,5.8,2.4Hz,1H),4.53-4.03(m,4H),3.32(s,1H),3.18-3.08(m,1H),3.06(s,1 H),1.94-1.65(m,4H),1.65-1.52(m,1H),1.47(s,6H),1.36-0.90(m,5H).MS:(ES)m / z C 30 H 34 F3N4O3[M+H] + Calculated value: 555.3, actual value: 555.3.

[0171] Example 11: N-(1-(cyclopropylmethyl)-3-(2-oxo-2-((1-(pyrimidin-2-yl)cyclopropyl)amino)ethyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide [ka] Step a: To a solution of tert-butyl 3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-3-(2-ethoxy-2-oxoethyl)azetidine-1-carboxylate (600 mg, 1.2 mmol) in 4:1 THF / HO (5 mL) was added LiOH (240 mg, 6.0 mmol). The reaction mixture was stirred at room temperature overnight. The solvent was removed under reduced pressure and the residue was treated with 1N HCl. The mixture was then diluted with EtOAc and washed with HO. The organic layer was concentrated and the crude material was purified by silica gel column chromatography to give 2-(1-(tert-butoxycarbonyl)-3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)azetidin-3-yl)acetic acid. MS: (ES) m / z C 20 H 22 F2N3O6[M+H] + The calculated value was 438.1, and the measured value was 438.2.

[0172] Step b: To a solution of 2-(1-(tert-butoxycarbonyl)-3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)azetidin-3-yl)acetic acid (230 mg, 0.52 mmol) in DCM (2 mL) was added 1-(pyrimidin-2-yl)cyclopropan-1-amine dihydrochloride (110 mg, 0.52 mmol), DIPEA (0.36 mL, 2.08 mmol) and HATU (220 mg, 0.58 mmol). The reaction mixture was stirred at room temperature for 1 h. The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography to give tert-butyl 3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-3-(2-oxo-2-((1-(pyrimidin-2-yl)cyclopropyl)amino)ethyl)azetidine-1-carboxylate. MS: (ES) m / z C 27 H 29 F2N6O5[M+H] + Calculated value: 555.2, measured value: 555.2.

[0173] Step c: A mixture of tert-butyl 3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamide)-3-(2-oxo-2-((1-(pyrimidin-2-yl)cyclopropyl)amino)ethyl)azetidine-1-carboxylate (288 mg, 0.52 mmol) in 1:1 DCM / TFA (2 mL) was stirred at room temperature for 1 h. The solvent was removed under reduced pressure to give 5-(2,4-difluorophenyl)-N-(3-(2-oxo-2-((1-(pyrimidin-2-yl)cyclopropyl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0174] step d: To a mixture of 5-(2,4-difluorophenyl)-N-(3-(2-oxo-2-((1-(pyrimidin-2-yl)cyclopropyl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide (57 mg, 0.1 mmol), triethylamine (0.05 mL, 0.4 mmol) and cyclopropanecarbaldehyde (15 mg, 0.2 mmol) in 4:1 DCM / MeOH (2 mL) was added NaBH(OAc)3 (42 mg, 0.2 mmol). The contents were stirred at room temperature for 1 hour, then filtered and purified by preparative HPLC to give N-(1-(cyclopropylmethyl)-3-(2-oxo-2-((1-(pyrimidin-2-yl)cyclopropyl)amino)ethyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide. 1 H NMR(400MHz,CD3OD)δ 8.60(dd,J=5.1,5.1Hz,2H),8.05(dd,J=6.4,6.4Hz,1H),7.31-7.17(m,3H ),7.12(dd,J=11.9,3.5Hz,1H),4.69(d,J=10.6Hz,2H),4.56(d,J=12.4Hz, 2H),3.25(d,J=7.3Hz,1H),3.20-3.09(m,3H),1.66(q,J=4.4Hz,2H),1.31 (dd,J=3.9,3.9Hz,2H),1.04(s,1H),0.70(s,2H),0.41(s,2H).MS:(ES)m / z C 26 H 27 F2N6O3[M+H]+ Calculated value: 509.2, measured value: 509.2.

[0175] Example 12: 5-(2,4-difluorophenyl)-N-(1-((3-methyloxetan-3-yl)methyl)-3-(2-oxo-2-((2-(pyridin-2-yl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] Step a: To a mixture of ethyl acetate 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)azetidin-3-yl) (600 mg, 1.29 mmol), triethylamine (0.55 mL, 4.0 mmol) and 3-methyloxetane-3-carbaldehyde (260 mg, 2.0 mmol) in 4:1 DCM / MeOH (10 mL) was added NaBH(OAc) (551 mg, 2.0 mmol). After stirring at room temperature for 1 h, the reaction was quenched with saturated NaHCO(aq) and the aqueous layer was extracted with DCM. The organic layers were combined, dried over MgSO, filtered and concentrated in vacuo. The crude residue was purified by silica gel column chromatography to give ethyl acetate 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-((3-methyloxetan-3-yl)methyl)azetidin-3-yl). MS: (ES) m / z C 22 H 26 F2N3O5[M+H] + Calculated value: 450.2, actual value: 450.2.

[0176] Step b: To a mixture of ethyl acetate 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-((3-methyloxetan-3-yl)methyl)azetidin-3-yl) (400 mg, 0.89 mmol) in 4:1 THF / HO (5 mL) was added LiOH (224 mg, 5.34 mmol). The reaction mixture was stirred at room temperature overnight. The volatiles were removed in vacuo and the residue was treated with 1N HCl. The solid was filtered, washed with HO and collected to give 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-((3-methyloxetan-3-yl)methyl)azetidin-3-yl)acetic acid. MS: (ES) m / z C 20 H 22 F2N3O5[M+H] + Calculated value: 422.1, measured value: 422.1.

[0177] Step c: To a solution of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-((3-methyloxetan-3-yl)methyl)azetidin-3-yl)acetic acid (45 mg, 0.10 mmol) in DCM (2 mL) was added 2-(pyridin-2-yl)propan-2-amine dihydrochloride (23 mg, 0.10 mmol), DIPEA (0.07 mL, 0.40 mmol) and HATU (46 mg, 0.12 mmol). The reaction mixture was stirred for 1 h and then concentrated under reduced pressure. The crude residue was purified by preparative HPLC to give 5-(2,4-difluorophenyl)-N-(1-((3-methyloxetan-3-yl)methyl)-3-(2-oxo-2-((2-(pyridin-2-yl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide. 1H NMR(400MHz,CD3OD)δ 8.59(s,1H),8.27(s,1H),8.06(ddd,J=8.5,8.5,6.2Hz,1H),7.89(s,1H),7.70(s,1H),7.32-7.17(m,2H),7.12(s,1H),4.7 6-4.47(m,6H),4.36(d,J=6.3Hz,2H),3.68(d,J=20.0Hz,2H),3.22(d,J=20.0Hz,2H),1.70(s,6H),1.39(s,3H).MS:(ES)m / z C 28 H 32 F2N5O4[M+H] + The calculated value was 540.2, and the actual value was 540.3.

[0178] Example 13: 5-(2,4-difluorophenyl)-N-(3-(2-((2-(2,6-dimethylpyridin-4-yl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (60 mg, 0.13 mmol) in DMF (2 mL) was added 2-(2,6-dimethylpyridin-4-yl)propan-2-amine (22 mg, 0.13 mmol), DIPEA (0.06 mL, 0.33 mmol) and HATU (61 mg, 0.16 mmol). The contents were stirred for 1 h and then concentrated under reduced pressure. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(3-(2-((2-(2,6-dimethylpyridin-4-yl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0179] Isomer 1: 1H NMR(400MHz,CD3OD)δ 8.05-8.00(m,1H),7.63(s,2H),7.30-7.11(m,3H),4.61-4.38(m,4H),3.24-3.18(m ,2H),2.68(s,6H),2.01-1.96(m,2H),1.75-1.48(m,13H),1.22(s,3H).MS:(ES)m / z C 32 H 40 F2N5O4[M+H] + Calculated value: 596.3, actual value: 596.5.

[0180] Isomer 2: 1 H NMR(400MHz,CD3OD)δ 8.08-8.00(m,1H),7.63-7.61(m,2H),7.30-7.12(m,3H),4.58-4.37(m,4H),3.30-3.1 9(m,2H),2.67(s,6H),1.82-1.73(m,13H),1.48-1.38(m,2H),1.21(s,3H).MS:(ES)m / z C 32 H 40 F2N5O4[M+H] + Calculated value: 596.3, actual value: 596.5.

[0181] Example 14: 5-(2,4-difluorophenyl)-N-(1-isobutyl-3-(2-((2-(2-methylpyridin-4-yl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] Step a: To a mixture of ethyl acetate 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)azetidin-3-yl) (300 mg, 0.8 mmol), DIPEA (0.29 mL, 1.6 mmol), acetic acid (0.19 mL, 3.2 mmol) and isobutraldehyde (118 mg, 1.6 mmol) in 2:1 DCM / MeOH (9 mL) was added NaBH3CN (52 mg, 0.8 mmol). The contents were stirred at room temperature for 1 h and the reaction was quenched with saturated NaHCO3(aq) and extracted with DCM. The combined organic layers were dried over MgSO4, filtered and concentrated in vacuo. The crude residue was purified by silica gel column chromatography to give ethyl acetate 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-isobutylazetidin-3-yl). MS: (ES) m / z C 21 H 26 F2N3O4[M+H] + Calculated value: 422.2, measured value: 422.2.

[0182] Step b: To a mixture of ethyl acetate 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-isobutylazetidin-3-yl) (164 mg, 0.4 mmol) in 1:1:1 THF / HO / MeOH (9 mL) was added LiOH (49 mg, 1.2 mmol). The mixture was stirred at room temperature for 2 h, concentrated in vacuo, and subsequently treated with 1N HCl (10 mL) and concentrated in vacuo to give 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-isobutylazetidin-3-yl)acetic acid. MS: (ES) m / z C 19 H 22 F2N3O4[M+H] + Calculated value: 394.2, actual value: 394.2.

[0183] Step c: To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamide)-1-isobutylazetidin-3-yl)acetic acid (55 mg, 0.14 mmol) in DMF (2 mL) was added 2-(2-methylpyridin-4-yl)propan-2-amine (21 mg, 0.14 mmol), DIPEA (0.06 mL, 0.35 mmol) and HATU (64 mg, 0.17 mmol). The contents were stirred for 1 h, filtered and subsequently purified by preparative HPLC to give 5-(2,4-difluorophenyl)-N-(1-isobutyl-3-(2-((2-(2-methylpyridin-4-yl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide. 1 H NMR(400MHz,DMSO-d6)δ 9.56(d,J=4.6Hz,1H),8.72-8.69(m,1H),8.48-8.44(m,1H),8.15-8.07 (m,1H),7.65-7.53(m,2H),7.47-7.37(m,1H),7.35-7.28(m,1H),7.27-7 .23(m,1H),4.56-4.39(m,2H),4.35-4.22(m,2H),3.15-2.99(m,4H),2.5 1(s,3H),1.89-1.78(m,1H),1.51(s,6H),0.89-0.86(m,6H).MS:(ES)m / z C 28 H 34 F2N5O3[M+H] + Calculated value: 526.3, actual value: 526.3.

[0184] Example 15: N-(3-(2-((2-(3-chlorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-isobutylazetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide [ka] To a solution of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamide)-1-isobutylazetidin-3-yl)acetic acid (40 mg, 0.10 mmol) in DMF (2 mL) was added 2-(3-chlorophenyl)propan-2-amine (17 mg, 0.10 mmol), DIPEA (0.04 mL, 0.25 mmol) and HATU (46 mg, 0.12 mmol). The mixture was stirred for 1 h, filtered and subsequently purified by preparative HPLC to give N-(3-(2-((2-(3-chlorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-isobutylazetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide. MS: (ES) m / z C 28 H 32 ClF2N4O3[M+H] + Calculated value: 545.2, actual value: 545.2.

[0185] Example 16: 5-(2,4-difluorophenyl)-N-(3-(2-oxo-2-((2-(pyridin-2-yl)propan-2-yl)amino)ethyl)-1-(2-oxaspiro[3.5]nonan-7-yl)azetidin-3-yl)isoxazole-3-carboxamide [ka] Step a: To a mixture of 5-(2,4-difluorophenyl)-N-(3-(2-oxo-2-((2-(pyridin-2-yl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide (70 mg, 0.1 mmol), DIPEA (0.04 mL, 0.2 mmol), acetic acid (0.03 mL, 0.4 mmol) and 2-oxaspiro[3.5]nonan-7-one (30 mg, 0.2 mmol) in 2:1 DCM / MeOH (3 mL) was added NaBH3CN (7 mg, 0.1 mmol). The contents were stirred at room temperature for 1 h, then the reaction was quenched with saturated NaHCO3(aq) and extracted with DCM. The combined organic layers were dried over MgSO4, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography to give 5-(2,4-difluorophenyl)-N-(3-(2-oxo-2-((2-(pyridin-2-yl)propan-2-yl)amino)ethyl)-1-(2-oxaspiro[3.5]nonan-7-yl)azetidin-3-yl)isoxazole-3-carboxamide. 1 H NMR (400MHz, CD3OD) δ 8.39(dd,J=4.9,1.8Hz,1H),8.07-8.01(m,1H),7.69-7.64(m,1H),7.41(d,J=8 .1Hz,1H),7.30-7.14(m,3H),7.07(d,J=3.5Hz,1H),4.42(s,2H),4.34(s,2H), 3.77-3.66(m,4H),3.05(s,2H),2.39-2.30(m,1H),2.15(d,J=13.5Hz,2H),1.7 9-1.75(m,2H),1.60(s,6H),1.50-1.43(m,2H),1.05-0.95(m,2H).MS:(ES)m / z C 31 H 36 F2N5O4[M+H] + Calculated value: 580.3, actual value: 580.2.

[0186] Example 17: 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-((2-(2-methylpyridin-4-yl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a mixture of 5-(2,4-difluorophenyl)-N-(3-(2-((2-(2-methylpyridin-4-yl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide (65 mg, 0.10 mmol), EtN (0.05 mL, 0.4 mmol) and 4-hydroxy-4-methylcyclohexan-1-one (26 mg, 0.20 mmol) in 4:1 DCM / MeOH (2 mL) was added NaBH(OAc) (43 mg, 0.20 mmol). The contents were stirred at room temperature for 1 h and then concentrated under reduced pressure. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-((2-(2-methylpyridin-4-yl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0187] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.50(d,J=6.3Hz,1H),8.05(ddd,J=8.5,8.5,6.0Hz,1H),7.84(s,1H),7.77(d,J=5.6Hz,1H),7.24(ddd,J=22.5,8.7,8.7,2.6Hz,2H),7.11(d ,J=4.0Hz,1H),4.60-4.35(m,4H),3.30-3.10(m,3H),2.75(s,3H),1.98 (s,2H),1.74-1.61(m,8H),1.56-1.36(m,4H),1.21(s,3H).MS:(ES)m / z C 31 H 38 F2N5O4[M+H] + Calculated value: 582.3, actual value: 582.3.

[0188] Isomer 2: 1 H NMR(400MHz,CD3OD)δ 8.50(d,J=6.2Hz,1H),8.06(ddd,J=8.6,8.6,6.2Hz,1H),7.83(d,J=8.0Hz,1H),7.77(s,1H),7.32-7.17(m,2H),7.13(s,1H),4.57-4. 35(m,4H),3.38-3.10(m,3H),2.72(d,J=4.5Hz,3H),1.85-1.70(m,4H),1.68-1.50(m,8H),1.48-1.36(m,2H),1.20(s,3H).MS:(ES)m / z C 31 H 37 F2N5O4[M+H] + Calculated value: 582.3, actual value: 582.3.

[0189] Example 18: 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-((2-(4-methylpyrimidin-2-yl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a mixture of 5-(2,4-difluorophenyl)-N-(3-(2-((2-(4-methylpyrimidin-2-yl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide (70 mg, 0.14 mmol), EtN (0.05 mL, 0.4 mmol) and 4-hydroxy-4-methylcyclohexan-1-one (36 mg, 0.28 mmol) in 4:1 DCM / MeOH (2 mL) was added NaBH(OAc) (60 mg, 0.28 mmol). The contents were stirred at room temperature for 1 h and then concentrated in vacuo. The residue was purified by preparative HPLC to give 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-((2-(4-methylpyrimidin-2-yl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide. 1 H NMR(400MHz,CD3OD)δ 8.49(dd,J=11.3,5.2Hz,1H),8.05(s,1H),7.30-7.06(m,4H),4.59-4.38(m,4H),3.14-3.06(m,3H),2.55-2.42(m,3 H),1.98(s,2H),1.75-1.56(m,2H),1.63(s,3H),1.62(s,3H),1.56-1.30(m,4H),1.20(d,J=8.0Hz,3H).MS:(ES)m / z C 30 H 37 F2N6O4[M+H] + Calculated value: 583.3, actual value: 583.3.

[0190] Example 19: 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-((2-(2-methoxypyridin-4-yl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DCM (1 mL) was added 2-(2-methoxypyridin-4-yl)propan-2-amine (17 mg, 0.10 mmol), DIPEA (0.052 mL, 0.3 mmol) and HATU (40 mg, 0.11 mmol). The contents were stirred for 1 h and then concentrated under reduced pressure. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-((2-(2-methoxypyridin-4-yl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0191] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.08(d,J=8.4Hz,1H),7.95(dd,J=17.0,6.0Hz,1H),7.25(dt,J=17.1,9.1Hz,2H),7 .16-7.09(m,1H),6.96(dd,J=19.8,5.9Hz,1H),6.80(d,J=18.6Hz,1H),4.58-4.44( m,3H),4.39(d,J=11.3Hz,1H),3.86(s,3H),3.31-3.20(m,1H),3.16(d,J=10.7Hz,2 H),1.97(s,2H),1.57(s,6H),1.70-1.24(m,6H),1.20(d,J=1.6Hz,3H).MS:(ES)m / z C 31 H 38 F2N5O5[M+H] + Calculated value: 598.3, actual value: 598.3.

[0192] Isomer 2: 1H NMR(400MHz,CD3OD)δ 8.12-7.97(m,2H),7.31-7.07(m,4H),7.00(d,J=19.9Hz,1H),4.57-4.42(m,3H),4.37(d,J=11.7Hz,1H),3.97(s,3H),3. 30-3.10(m,3H),1.80-1.68(m,4H),1.65-1.50(m,2H),1.59(s,6H),1.50-1.36(m,2H),1.20(d,J=3.6Hz,3H).MS:(ES)m / z C 31 H 38 F2N5O5[M+H] + Calculated value: 598.3, actual value: 598.3.

[0193] Example 20: 5-(2,4-difluorophenyl)-N-(3-(2-((2-(4,6-dimethylpyrimidin-2-yl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] Step a: To a solution of 4,6-dimethylpyrimidine-2-carbonitrile (1.34 g, 10 mmol) in ether (35 mL) at room temperature was added a solution of 3.0 M methylmagnesium bromide (10 mL, 30 mmol) dropwise, followed by titanium(IV) isopropoxide (3.04 mL, 10 mmol). The reaction mixture was stirred at 40° C. overnight and then quenched with a solution of H2O followed by 10% NaOH(aq). The contents were diluted with DCM and filtered through a plug of Celite. The filtrate was concentrated and the crude material was purified by silica gel column chromatography to give 2-(4,6-dimethylpyrimidin-2-yl)propan-2-amine.

[0194] Step b: To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (60 mg, 0.12 mmol) in DCM (2 mL) was added 2-(4,6-dimethylpyrimidin-2-yl)propan-2-amine (21 mg, 0.12 mmol), DIPEA (0.04 mL, 0.24 mmol) and HATU (52 mg, 0.14 mmol). The contents were stirred for 1 h and then concentrated under reduced pressure. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(3-(2-((2-(4,6-dimethylpyrimidin-2-yl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0195] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.07-7.98(m,1H),7.23(ddd,J=18.7,9.3,9.3Hz,2H),7.13-7.06(m,1H),7.04(s,1H),5.49(s,1H),4.58-4.32(m,4H),3. 50-3.20(m,1H),3.11(d,J=9.8Hz,2H),2.40(s,6H),1.99(s,2H),1.75-1.32(m,6H),1.62(s,6H),1.20(s,3H).MS:(ES)m / z C 31 H 39 F2N6O4[M+H] + Calculated value: 597.3, actual value: 597.3.

[0196] Isomer 2: 1H NMR(400MHz,CD3OD)δ 8.07-7.98(m,1H),7.23(ddd,J=18.7,9.3,9.3Hz,2H),7.14-7.06(m,1H),7.04(s,1H),4.58-4.38(m,4H),3.30-3.10 (m,3H),2.40(s,6H),1.82-1.70(m,4H),1.62(s,6H),1.62-1.50(m,2H),1.48-1.36(m,2H),1.21(s,3H).MS:(ES)m / z C 31 H 39 F2N6O4[M+H] + Calculated value: 597.3, actual value: 597.3.

[0197] Example 21: 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2-((2-(6-(trifluoromethyl)pyridin-2-yl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (60 mg, 0.12 mmol) in DMF (1 mL) was added 2-(6-(trifluoromethyl)pyridin-2-yl)propan-2-amine (25 mg, 0.12 mmol), DIPEA (0.052 mL, 0.3 mmol) and HATU (52 mg, 0.14 mmol). The contents were stirred for 1 h and then concentrated under reduced pressure. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2-((2-(6-(trifluoromethyl)pyridin-2-yl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0198] Isomer 1: 1H NMR(400MHz,CD3OD)δ 8.06(d,J=8.1Hz,1H),7.87(ddd,J=15.4,7.9,7.9Hz,1H),7.68-7.54(m,2H),7.31-7.17(m,2H),7.14-7.06(m,1H),4.56-4. 30(m,4H),3.40-3.20(m,1H),3.15(d,J=13.0Hz,2H),1.96(s,2H),1.63(s,6H),1.74-1.30(m,6H),1.19(s,3H).MS:(ES)m / z C 31 H 35 F5N5O4[M+H] + Calculated value: 636.3, measured value: 636.3.

[0199] Isomer 2: 1 H NMR(400MHz,CD3OD)δ 8.06(d,J=8.5Hz,1H),7.87(ddd,J=8.4,8.4,8.4Hz,1H),7.69-7.52(m,2H),7.24(ddd,J=17.8,9.3,9.3Hz,2H),7.10(dd,J=10. 7,3.5Hz,1H),4.56-4.30(m,4H),3.40-3.10(m,3H),1.85-1.70(m,4H),1.62(s,6H),1.62-1.35(s,4H),1.19(s,3H).MS:(ES)m / z C 31 H 35 F5N5O4[M+H] + Calculated value: 636.3, measured value: 636.3.

[0200] Example 22: 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2-((2-(3-(trifluoromethoxy)phenyl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (45 mg, 0.093 mmol) in DMF (1.5 mL) was added 2-(3-(trifluoromethoxy)phenyl)propan-2-amine (65 mg, 0.30 mmol), HATU (57 mg, 0.15 mmol) and DIPEA (0.061 mL, 0.35 mmol). The contents were stirred for 1 h, quenched with water, and purified by preparative HPLC to give the two isomers of 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2-((2-(3-(trifluoromethoxy)phenyl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0201] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.58(s,1H),8.49(s,1H),8.02-8.10(m,1H),7.15-7.34(m,5H),7.03-7.14(m,2H),4.54(d,J=11.6Hz,1H),4.66(s,2H),4. 37(d,J=11.6Hz,1H),3.22-3.40(m,1H),3.15(d,J=12.4Hz,2H),1.96(bs,2H),1.30-1.56(m,12H),1.20(s,3H).MS:(ES)m / z C 32 H 36 F5N4O5[M+H] + Calculated value: 651.3, actual value: 651.5.

[0202] Isomer 2: 1H NMR(400MHz,CD3OD)δ 8.52(s,1H),8.49(s,1H),8.02-8.11(m,1H),7.15-7.40(m,5H),7.03-7.14(m,2H),4.42-4.54(m,3H),4.36(d,J= 12.4Hz,1H),3.10-3.35(m,3H),1.70-1.90(m,4H),1.50-1.62(m,8H),1.34-1.48(m,2H),1.20(s,3H).MS:(ES)m / z C 32 H 36 F5N4O5[M+H] + Calculated value: 651.3, actual value: 651.5.

[0203] Example 23: N-(3-(2-((2-(3,5-dichlorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide [ka] To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (30 mg, 0.062 mmol) in DMF (1.5 mL) was added 2-(3,5-dichlorophenyl)propan-2-amine (25 mg, 0.12 mmol), HATU (30 mg, 0.079 mmol) and EtN (0.040 mL, 0.28 mmol). The mixture was stirred for 0.5 h, quenched with water and purified by preparative HPLC to give N-(3-(2-((2-(3,5-dichlorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide. 1H NMR(400MHz,CD3OD)δ 8.61(s,1H),8.51(s,1H),8.00-8.10(m,1H),7.15-7.32(m,5H),7.06-7.13(m,1H),4.54(d,J=12.4Hz,1H),4.47(s,2H),4. 39(d,J=12.0Hz,1H),3.22-3.40(m,1H),3.15(d,J=12.4Hz,2H),1.97(bs,2H),1.30-1.75(m,12H),1.20(s,3H).MS:(ES)m / z C 31 H 35 Cl2F2N4O4[M+H] + Calculated value: 635.2, actual value: 635.5.

[0204] Example 24: 5-(2,4-difluorophenyl)-N-(3-(2-((2-(2-ethylpyridin-4-yl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] Step a: To a flask containing 2-ethylisonicotinonitrile (1.00 g, 7.56 mmol) in ether (50 mL) under N2 was added a 3.0 M solution of methylmagnesium bromide (7.56 mL, 22.7 mmol) and titanium(IV) isopropoxide (8.82 mL, 29.8 mmol). The mixture was stirred at 40° C. overnight, cooled to 0° C., quenched with water, and diluted with a solution of 10% NaOH(aq). The contents were filtered through Celite and rinsed with DCM. The organic layer of the filtrate was dried over NaSO4, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to give 2-(2-ethylpyridin-4-yl)propan-2-amine. MS: (ES) m / z C 10 H 17 N2[M+H] + The calculated value was 165.1, and the actual value was 165.0.

[0205] Step b: To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (45 mg, 0.093 mmol) in DMF (1.5 mL) was added 2-(2-ethylpyridin-4-yl)propan-2-amine (25 mg, 0.15 mmol), HATU (45 mg, 0.12 mmol) and EtN (0.052 mL, 0.37 mmol). The contents were stirred for 1 h, quenched with water, and purified by preparative HPLC to give the two isomers of 5-(2,4-difluorophenyl)-N-(3-(2-((2-(2-ethylpyridin-4-yl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0206] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.85(s,1H),8.83(s,1H),8.52(d,J=6.0Hz,1H),8.02-8.10(m,1H),7.86(s,1H) ,7.80(s,1H),7.18-7.31(m,2H),7.12(s,1H),4.36-4.63(m,4H),3.26-3.40(m, 1H),3.16-3.26(m,2H),2.97-3.08(m,2H),1.93-2.02(m,2H),1.66-1.74(m,2H) ,1.64(s,6H),1.40-1.56(m,4H),1.35(t,J=7.6Hz,3H),1.21(s,3H).MS:(ES)m / z C 32 H 40 F2N5O4[M+H] + Calculated value: 596.3, measured value: 596.4.

[0207] Isomer 2: 1H NMR(400MHz,CD3OD)δ 8.85(s,1H),8.81(s,1H),8.52(d,J=6.0Hz,1H),8.02-8.10(m,1H),7.7 4-7.87(m,2H),7.18-7.31(m,2H),7.12(s,1H),4.34-4.60(m,4H),3.16- 3.26(m,1H),3.18(s,2H),2.98-3.04(m,2H),1.72-1.84(m,4H),1.64(s ,6H),1.37-1.63(m,4H),1.34(t,J=7.4Hz,3H),1.20(s,3H).MS:(ES)m / z C 32 H 40 F2N5O4[M+H] + Calculated value: 596.3, measured value: 596.4.

[0208] Example 25: 5-(2,4-difluorophenyl)-N-(1-isobutyl-3-(2-oxo-2-((2-(pyridin-2-yl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a mixture of 5-(2,4-difluorophenyl)-N-(3-(2-oxo-2-((2-(pyridin-2-yl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide (35 mg, 0.071 mmol), isobutyraldehyde (30 mg, 0.042 mmol), EtN (0.050 mL, 0.35 mmol) in DCM (2 mL) was added NaBH(OAc) (50 mg, 0.23 mmol). The contents were stirred for 1 h, quenched with water, concentrated and purified by preparative HPLC to give 5-(2,4-difluorophenyl)-N-(1-isobutyl-3-(2-oxo-2-((2-(pyridin-2-yl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide. 1H NMR(400MHz,CD3OD)δ 8.64(ddd,J=7.6,2.0,1.2Hz,1H),8.43(dd,J=10.4,10.4Hz,1H),7.97-8.10(m,2H),7.83(ddd,J=8.8,8.8,1.6Hz,1H),7.16-7.29(m ,2H),7.09(d,J=4.8Hz,1H),4.25-4.75(m,4H),3.00-3.40(m,4H),1.85-2.00(m,1H),1.72(s,6H),0.97(d,J=9.2Hz,6H).MS:(ES)m / z C 27 H 32 F2N5O3[M+H] + Calculated value: 512.2, measured value: 512.4.

[0209] Example 26: N-(3-(2-((2-(1H-pyrrolo[2,3-b]pyridin-4-yl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide [ka] Step a: To a flask containing 1H-pyrrolo[2,3-b]pyridine-4-carbonitrile (1.00 g, 7.0 mmol) in ether (50 mL) and THF (50 mL) under N2 was added a solution of 3.0 M methylmagnesium bromide (7.0 mL, 21 mmol) followed by titanium(IV) isopropoxide (2.07 mL, 7.0 mmol). The mixture was stirred at 40° C. overnight, cooled to 0° C., quenched with water and diluted with a solution of 10% NaOH(aq). The contents were filtered through Celite and rinsed with DCM. The organic layer of the filtrate was dried over NaSO4, concentrated under reduced pressure and purified by silica gel column chromatography to give 2-(1H-pyrrolo[2,3-b]pyridin-4-yl)propan-2-amine. MS: (ES) m / z C 10 H 14 N3[M+H] + Calculated value: 176.1, actual value: 176.1.

[0210] Step b: To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (35 mg, 0.072 mmol) in DMF (1.5 mL) was added 2-(1H-pyrrolo[2,3-b]pyridin-4-yl)propan-2-amine (25 mg, 0.14 mmol), HATU (30 mg, 0.079 mmol) and EtN (0.040 mL, 0.28 mmol). The contents were stirred for 0.5 h, quenched with water and purified by preparative HPLC to give N-(3-(2-((2-(1H-pyrrolo[2,3-b]pyridin-4-yl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide. 1 H NMR(400MHz,CD3OD)δ 9.03(s,1H),8.94(s,1H),8.27(d,J=5.2Hz,1H),8.08(d,J=6.8Hz,1H),7.41(d,J=6.0Hz,1H),7.15-7.32(m,4H),7.07(s,1H),6. 81(d,J=14.8Hz,1H),4.22-4.49(m,4H),3.12-3.30(m,3H),1.89(s,2H),1.76(s,6H),1.25-1.71(m,6H),1.17(s,3H).MS:(ES)m / z C 32 H 37 F2N6O4[M+H] + Calculated value: 607.3, measured value: 607.5.

[0211] Example 27: 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-((2-(4-methylpyridin-2-yl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (0.075 g, 0.17 mmol) in DCM (1 mL) was added 2-(4-methylpyridin-2-yl)propan-2-amine (0.082 g, 0.55 mmol), DIPEA (0.09 mL, 0.50 mmol) and HATU (0.130 g, 0.33 mmol). The mixture was stirred at room temperature overnight and then concentrated under reduced pressure. The residue was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-((2-(4-methylpyridin-2-yl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0212] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.40(s,1H),8.05(m,1H),7.75(s,1H),7.59(d,J=6.0Hz,1H),7.34-7.16(m,2H),7.11(s,1H),4.58-4.34(m,3H),3.21-3.11(m,3H) ),2.55(s,3H),2.02-1.92(m,2H),1.82-1.72(m,1H),1.68(s,6H),1.58-1.45(m,5H),1.44-1.32(m,1H),1.21(s,3H).MS:(ES)m / z C 31 H 38 F2N5O4[M+H] + Calculated value: 582.3, actual value: 582.5.

[0213] Isomer 2: 1H NMR(400MHz,CD3OD)δ 8.40(s,1H),8.05(m,1H),7.76(s,1H),7.61(d,J=6.0Hz,1H),7.30-7.18(m,2H),7.13-7.08(m,1H),4.58-4.31(m,4H),3.2 2-3.07(m,3H),2.55(s,3H),1.85-1.70(m,3H),1.68(s,6H),1.63-1.49(m,2H),1.47-1.36(m,3H),1.20(s,3H).MS:(ES)m / z C 31 H 38 F2N5O4[M+H] + Calculated value: 582.3, actual value: 582.5.

[0214] Example 28: 5-(2,4-difluorophenyl)-N-(3-(2-((2-(4,6-dimethylpyridin-2-yl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a solution of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (0.075 g, 0.15 mmol) in DMF (1 mL) was added 2-(4,6-dimethylpyridin-2-yl)propan-2-amine (0.033 g, 0.20 mmol), HATU (0.129 g, 0.34 mmol) and DIPEA (0.09 mL, 0.50 mmol). The reaction mixture was stirred at room temperature for 5 h and concentrated under reduced pressure. The crude material was purified by preparative HPLC to give 5-(2,4-difluorophenyl)-N-(3-(2-((2-(4,6-dimethylpyridin-2-yl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide. 1H NMR(400MHz,CD3OD)δ 8.08-8.00(m,1H),7.69(s,1H),7.57(s,1H),7.34-7.16(m,2H),7.10(s,1H),4.65-4.35(m,4H),3.40-3.15(m,2H),2.70- 2.60(m,1H),2.57(s,6H),2.05-1.90(m,2H),1.75-1.70(m,1H),1.70(s,6H),1.57-1.33(m,5H),1.21(s,3H).MS:(ES)m / z C 32 H 40 F2N5O4[M+H] + Calculated value: 596.3, actual value: 596.5.

[0215] Example 29: 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2-((2-(pyrimidin-2-yl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a solution of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (0.06 g, 0.13 mmol) in DMF (1 mL) was added 2 2-(pyrimidin-2-yl)propan-2-amine dihydrochloride (0.06 g, 0.29 mmol), DIPEA (0.07 mL, 0.40 mmol) and HATU (0.10 g, 0.26 mmol). The reaction mixture was stirred at room temperature overnight and then concentrated under reduced pressure. The aqueous layer was extracted with EtOAc and the organic layers were combined, dried over sodium sulfate, filtered and concentrated. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2-((2-(pyrimidin-2-yl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0216] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.68(dd,J=9.2,4.9Hz,2H),8.15-7.96(m,1H),7.32-7.27(m,2H),7.27-7 .16(m,1H),7.11(dd,J=8.1,3.5Hz,1H),4.58-4.37(m,4H),3.27-3.20(m,1 H),3.09(d,J=16.3Hz,2H),1.97(bs,2H),1.75-1.67(m,2H),1.64(s,3H),1 .63(s,3H),1.55-1.45(m,2H),1.43-1.32(m,2H),1.21(s,3H).MS:(ES)m / z C 29 H 35 F2N6O4[M+H] + Calculated value: 569.3, measured value: 569.4.

[0217] Isomer 2: 1 H NMR(400MHz,CD3OD)δ 8.67(dd,J=7.5,4.9Hz,2H),8.05(m,1H),7.33-7.26(m,2H),7.26-7.16(m,1H),7.11(dd,J=10.7,3.4Hz,1H),4.56-4.36(m,4H) ,3.19-3.05(m,3H),1.87-1.70(m,4H),1.63(s,3H),1.62(s,3H),1.61-1.51(m,2H),1.48-1.35(m,2H),1.21(s,3H).MS:(ES)m / z C 29 H 35 F2N6O4[M+H] + Calculated value: 569.3, measured value: 569.4.

[0218] Example 30: 5-(2,4-difluorophenyl)-N-(1-(2,2-dimethylcyclohexyl)-3-(2-oxo-2-((2-(pyridin-2-yl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] Step a: To a mixture of ethyl acetate 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)azetidin-3-yl) (800 mg, 2.0 mmol), Et3N (0.55 mL, 4.0 mmol) and 2,2-dimethylcyclohexanone (505 mg, 4.0 mmol) in 4:1 DCM / MeOH (20 mL) was added NaBH(OAc)3 (840 mg, 4.0 mmol). The reaction mixture was stirred at room temperature for 1 h, then quenched with saturated NaHCO3(aq) and extracted with DCM. The combined organic layers were dried over MgSO4, filtered and concentrated in vacuo. The crude residue was purified by silica gel column chromatography to give ethyl acetate 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(2,2-dimethylcyclohexyl)azetidin-3-yl). MS: (ES) m / z C 25 H 32 F2N3O4[M+H] + The calculated value was 476.2, and the actual value was 476.5.

[0219] Step b: To a mixture of ethyl acetate 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(2,2-dimethylcyclohexyl)azetidin-3-yl) (600 mg, 1.2 mmol) in 1:1 THF / H2O (20 mL) was added NaOH (300 mg, 7.5 mmol). The mixture was stirred at room temperature overnight and then treated with 1N HCl and MeCN. The contents were concentrated under reduced pressure and the crude residue was triturated with acetone and filtered through Celite. The filtrate was concentrated to give ethyl 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(2,2-dimethylcyclohexyl)azetidin-3-yl)acetate. MS: (ES) m / z C 23 H 28 F2N3O4[M+H] + The calculated value was 448.2, and the measured value was 448.4.

[0220] Step c: To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(2,2-dimethylcyclohexyl)azetidin-3-yl)acetic acid (100 mg, 0.21 mmol) in DCM (2 mL) was added 2-(pyridin-2-yl)propan-2-amine dihydrochloride (43 mg, 0.21 mmol), DIPEA (0.14 mL, 0.82 mmol) and HATU (82 mg, 0.22 mmol). The contents were stirred for 1 h and then concentrated under reduced pressure. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(1-(2,2-methylcyclohexyl)-3-(2-oxo-2-((2-(pyridin-2-yl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0221] Isomer 1: 1 H NMR(400MHz,DMSO-d6)δ 9.81-9.50(m,2H),8.51-8.37(m,2H),8.17-8.06(m,1H),7.69-7.48(m,1H),7.41-7.10(m,4H),4.57-4.22(m,3 H),3.36-3.25(m,1H),3.17-3.01(m,2H),2.26-2.14(m,1H),1.76-1.10(m,14H),0.90-0.79(m,6H).MS:(ES)m / z C 31 H 38 F2N5O3[M+H] + Calculated value: 566.3, actual value: 566.3.

[0222] Isomer 2: 1H NMR(400MHz,DMSO-d6)δ 9.55(s,1H),9.40(s,1H),8.50-8.37(m,2H),8.18-8.03(m,1H),7.63-7.44(m,2H),7.42-7.10(m,3H),4.44-4.25(m,1H),3.18( MS:(ES)m / z C 31 H 38 F2N5O4[M+H] + Calculated value: 566.3, actual value: 566.3.

[0223] Example 31: 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2-((1-(pyrimidin-2-yl)cyclopropyl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 1-(pyrimidin-2-yl)cyclopropylamine dihydrochloride (42 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.105 mL, 0.60 mmol). The contents were stirred at room temperature for 1 h and then quenched with 3 drops of HO. The crude material was purified by preparative HPLC to give 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2-((1-(pyrimidin-2-yl)cyclopropyl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide. 1H NMR(400MHz,DMSO-d6)δ 9.57(s,1H),8.89(s,1H),8.57-8.50(m,2H),8.13-8.00(m,1H),7.65-7.54(m,1H),7.39-7.28(m,1H),7.25-7. 16(m,2H),4.48-4.35(m,4H),3.01-2.94(m,2H),1.79-1.38(m,9H),1.33-1.12(m,4H),1.08(s,3H).MS:(ES)m / z C 29 H 33 F2N6O4[M+H] + Calculated value: 567.3, actual value: 567.5.

[0224] Example 32: 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-((2-(2-methylphenyl)propan-2-yl)amino)-2-oxoethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-(o-tolyl)propan-2-amine (34 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The contents were stirred at room temperature for 1 h and then quenched with 3 drops of HO. The crude material was purified by preparative HPLC to give 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2-((2-(o-tolyl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide. 1H NMR(400MHz,CD3OD)δ 8.14-7.99(m,1H),7.34-6.88(m,7H),4.58-4.30(m,4H),3.25-3.03(m,3H),2.26-2.14( m,3H),1.90-1.68(m,4H),1.68-1.48(m,8H),1.48-1.31(m,2H),1.20(s,3H).MS:(ES)m / z C 32 H 39 F2N4O4[M+H] + Calculated value: 581.3, actual value: 581.3.

[0225] Example 33: N-(3-(2-((2-(2-chlorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide [ka] To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-(2-chlorophenyl)propan-2-amine (36 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The contents were stirred at room temperature for 1 h and then quenched with 3 drops of HO. The crude material was purified by preparative HPLC to give two separated isomers of N-(3-(2-((2-(2-chlorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide.

[0226] Isomer 1: 1H NMR(400MHz,CD3OD)δ 8.14-8.02(m,1H),7.56-7.48(m,1H),7.33-7.18(m,3H),7.18-7.04(m,3H),4.56-4.29(m,4H),3.27- 3.08(m,3H),2.00-1.88(m,2H),1.77-1.64(m,7H),1.59-1.26(m,5H),1.22-1.16(m,3H).MS:(ES)m / z C 31 H 36 ClF2N4O4[M+H] + The calculated value was 601.2, and the measured value was 601.3.

[0227] Isomer 2: 1 H NMR(400MHz,CD3OD)δ 8.13-8.02(m,1H),7.54-7.48(m,1H),7.32-7.18(m,3H),7.17-7.04(m,3H),4.58-4.25(m,4 H),3.24-3.04(m,3H),1.83-1.66(m,10H),1.63-1.33(m,4H),1.23-1.16(m,3H).MS:(ES)m / z C 31 H 36 ClF2N4O4[M+H] + The calculated value was 601.2, and the measured value was 601.3.

[0228] Example 34: 5-(2,4-difluorophenyl)-N-(3-(2-((2-(2-fluorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-(2-fluorophenyl)propan-2-amine (34 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The contents were stirred at room temperature for 1 h and then quenched with 3 drops of HO. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(3-(2-((2-(2-fluorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0229] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.13-8.03(m,1H),7.40-7.01(m,6H),6.89-6.73(m,1H),4.59-4.29(m,4H),3.27-3.07( m,3H),2.02-1.89(m,2H),1.74-1.55(m,8H),1.55-1.27(m,4H),1.20(s,3H).MS:(ES)m / z C 31 H 36 F3N4O4[M+H] + Calculated value: 585.3, actual value: 585.3.

[0230] Isomer 2: 1 H NMR(400MHz,CD3OD)δ 8.13-8.04(m,1H),7.38-7.01(m,6H),6.85-6.75(m,1H),4.54-4.30(m,4H),3.15-3.05(m,3H),1.88-1.32(m,14H),1.20(s,3H).MS:(ES)m / z C 31 H 36 F3N4O4[M+H] + Calculated value: 585.3, actual value: 585.3.

[0231] Example 35: 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2-((2-(m-tolyl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-(m-tolyl)propan-2-amine (34 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The contents were stirred at room temperature for 1 h and then quenched with 3 drops of HO. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(1-(4-hydroxy-4-methylcyclohexyl)-3-(2-oxo-2-((2-(m-tolyl)propan-2-yl)amino)ethyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0232] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.11-8.01(m,1H),7.32-7.18(m,2H),7.16-7.01(m,4H),6.98-6.90(m,1H),4.61-4.27(m,4H),3.39-3.18(m,1 H),3.16-3.07(m,2H),2.25-2.16(m,3H),2.02-1.88(m,2H),1.74-1.27(m,12H),1.24-1.16(m,3H).MS:(ES)m / z C 32 H 39 F2N4O4[M+H] + Calculated value: 581.3, actual value: 581.3.

[0233] Isomer 2: 1H NMR(400MHz,CD3OD)δ 8.14-7.99(m,1H),7.34-6.88(m,7H),4.58-4.30(m,4H),3.25-3.03(m,3H),2.26-2.14( m,3H),1.90-1.68(m,4H),1.68-1.48(m,8H),1.48-1.31(m,2H),1.20(s,3H).MS:(ES)m / z C 32 H 39 F2N4O4[M+H] + Calculated value: 581.3, actual value: 581.3.

[0234] Example 36: N-(3-(2-((2-(3-chloro-5-fluorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide [ka] Step a: To a solution of 3-chloro-5-fluorobenzonitrile (1.00 g, 6.37 mmol) in ether (100 mL) was added a 3.0 M solution of methylmagnesium bromide (7.30 mL, 21.9 mmol) dropwise, followed by titanium(IV) isopropoxide (2.16 mL, 7.30 mmol). The resulting mixture was refluxed under N2 overnight, then cooled to 0 °C and quenched with water, followed by a solution of 10% NaOH(aq). The contents were filtered through a plug of Celite and washed with DCM. The organic layer of the filtrate was collected, dried over Na2SO4, concentrated under reduced pressure and purified by silica gel column chromatography to give 2-(3-chloro-5-fluorophenyl)propan-2-amine. MS: (ES) m / z C9H 12 ClFN[M+H] + Calculated value: 188.1, actual value: 188.1.

[0235] Step b: To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-(3-chloro-5-fluorophenyl)propan-2-amine (38 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The contents were stirred at room temperature for 1 h and then quenched with 3 drops of HO. The crude residue was purified by preparative HPLC to give two separated isomers of N-(3-(2-((2-(3-chloro-5-fluorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-5-(2,4-difluorophenyl)isoxazole-3-carboxamide.

[0236] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.11-7.98(m,1H),7.30-7.07(m,4H),7.04-6.93(m,2H),4.58-4.33(m,4H),3.40-3.22(m,1H), 3.14(s,2H),2.04-1.91(m,2H),1.78-1.46(m,10H),1.46-1.29(m,2H),1.21(s,3H).MS:(ES)m / z C 31 H 35 ClF3N4O4[M+H] + Calculated value: 619.2, measured value: 619.2.

[0237] Isomer 2: 1 H NMR(400MHz,CD3OD)δ 8.10-7.99(m,1H),7.30-7.18(m,2H),7.15-7.07(m,2H),7.02-6.94(m,2H),4.57-4.32(m,4H),3. 15-3.10(m,3H),1.86-1.69(m,4H),1.59-1.55(m,8H),1.47-1.35(m,2H),1.20(s,3H).MS:(ES)m / z C 31 H 35 ClF3N4O4[M+H]+ Calculated value: 619.2, measured value: 619.2.

[0238] Example 37: 5-(2,4-difluorophenyl)-N-(3-(2-((2-(3-fluoro-5-methoxyphenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] Step a: To a solution of 3-fluoro-5-methoxybenzonitrile (1.00 g, 6.99 mmol) in ether (100 mL) was added a 3.0 M solution of methylmagnesium bromide (7.30 mL, 21.9 mmol) followed by dropwise addition of titanium(IV) isopropoxide (2.16 mL, 7.30 mmol). The resulting mixture was refluxed overnight under N2, then cooled to 0 °C and quenched with water followed by a solution of 10% NaOH(aq). The contents were filtered through a plug of Celite and washed with DCM. The organic layer of the filtrate was collected, dried over Na2SO4, concentrated under reduced pressure and purified by silica gel column chromatography to give 2-(3-fluoro-5-methoxyphenyl)propan-2-amine. MS: (ES) m / z C 10 H 15 FNO[M+H] + Calculated value: 184.1, actual value: 184.1.

[0239] Step b: To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-(3-fluoro-5-methoxyphenyl)propan-2-amine (38 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The contents were stirred at room temperature for 1 h and then quenched with 3 drops of HO. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(3-(2-((2-(3-fluoro-5-methoxyphenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0240] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.12-8.00(m,1H),7.31-7.17(m,2H),7.13-7.06(m,1H),6.65(s,1H),6.62-6.57(m,1H),6.49-6.44(m,1H),4.55-4.33(m,4 H),3.72-3.64(m,3H),3.26-3.05(m,3H),1.88-1.69(m,4H),1.66-1.48(m,8H),1.48-1.34(m,2H),1.20(s,3H).MS:(ES)m / z C 32 H 38 F3N4O5[M+H] + Calculated value: 615.3, measured value: 615.3.

[0241] Isomer 2: 1H NMR(400MHz,CD3OD)δ 8.12-8.00(m,1H),7.33-7.17(m,2H),7.14-7.06(m,1H),6.71-6.56(m,2H),6.52-6.43(m,1H),4.58-4.33(m,4H),3.7 2-3.64(m,3H),3.39-3.20(m,1H),3.17-3.08(m,2H),2.04-1.89(m,2H),1.74-1.28(m,12H),1.20(s,3H).MS:(ES)m / z C 32 H 38 F3N4O5[M+H] + Calculated value: 615.3, measured value: 615.3.

[0242] Example 38: 5-(2,4-difluorophenyl)-N-(3-(2-((2-(2-fluoro-5-methoxyphenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] Step a: To a solution of 2-fluoro-5-methoxybenzonitrile (1.00 g, 6.99 mmol) in ether (100 mL) was added a 3.0 M solution of methylmagnesium bromide (7.30 mL, 21.9 mmol) dropwise, followed by titanium(IV) isopropoxide (2.16 mL, 7.30 mmol). The resulting mixture was refluxed overnight under N2, then cooled to 0 °C and quenched with water, followed by a solution of 10% NaOH(aq). The mixture was filtered through a plug of Celite and washed with DCM. The organic layer of the filtrate was collected, dried over Na2SO4, concentrated under reduced pressure and purified by silica gel column chromatography to give 2-(2-fluoro-5-methoxyphenyl)propan-2-amine. MS: (ES) m / z C 10 H 15 FNO[M+H] + Calculated value: 184.1, actual value: 184.1.

[0243] Step b: To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-(2-fluoro-5-methoxyphenyl)propan-2-amine (38 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The reaction mixture was stirred at room temperature for 1 h and then quenched with 3 drops of HO. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(3-(2-((2-(2-fluoro-5-methoxyphenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0244] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.12-8.02(m,1H),7.32-7.18(m,2H),7.14-7.07(m,1H),6.88-6.81(m,1H),6.80-6.67(m,2H),4.57-4.3 2(m,4H),3.71(s,3H),3.28-3.02(m,3H),2.03-1.90(m,2H),1.75-1.27(m,12H),1.20(s,3H).MS:(ES)m / z C 32 H 38 F3N4O5[M+H] + Calculated value: 615.3, measured value: 615.3.

[0245] Isomer 2: 1 H NMR(400MHz,CD3OD)δ 8.12-8.03(m,1H),7.32-7.18(m,2H),7.14-7.07(m,1H),6.87-6.81(m,1H),6.74-6.67(m,2H),4.58-4.2 9(m,4H),3.70(s,3H),3.15-3.05(m,3H),1.85-1.70(m,4H),1.67-1.33(m,10H),1.20(s,3H).MS:(ES)m / z C 32H 38 F3N4O5[M+H] + Calculated value: 615.3, measured value: 615.3.

[0246] Example 39: 5-(2,4-difluorophenyl)-N-(3-(2-((2-(2-fluoro-5-methylphenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide [ka] Step a: To a solution of 2-fluoro-5-methylbenzonitrile (1.00 g, 7.87 mmol) in ether (100 mL) was added a 3.0 M solution of methylmagnesium bromide (7.30 mL, 21.9 mmol) dropwise, followed by titanium(IV) isopropoxide (2.16 mL, 7.30 mmol). The resulting mixture was refluxed overnight under N2, then cooled to 0 °C and quenched with water, followed by a solution of 10% NaOH(aq). The mixture was filtered through a plug of Celite and washed with DCM. The organic layer of the filtrate was collected, dried over Na2SO4, concentrated under reduced pressure and purified by silica gel column chromatography to give 2-(2-fluoro-5-methylphenyl)propan-2-amine. MS: (ES) m / z C 10 H 15 FN[M+H] + Calculated value: 168.1, actual value: 168.1.

[0247] Step b: To a mixture of 2-(3-(5-(2,4-difluorophenyl)isoxazole-3-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (50 mg, 0.10 mmol) in DMF (2 mL) was added 2-(2-fluoro-5-methylphenyl)propan-2-amine (38 mg, 0.20 mmol), HATU (77 mg, 0.20 mmol) and DIPEA (0.070 mL, 0.40 mmol). The contents were stirred at room temperature for 1 h and then quenched with 3 drops of HO. The crude material was purified by preparative HPLC to give the two separated isomers of 5-(2,4-difluorophenyl)-N-(3-(2-((2-(2-fluoro-5-methylphenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)isoxazole-3-carboxamide.

[0248] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.13-8.01(m,1H),7.35-7.17(m,2H),7.17-7.06(m,2H),7.05-6.91(m,1H),6.78-6.58(m,1H),4.57-4.30(m,4H),3.3 9-3.15(m,1H),3.15-2.96(m,2H),2.28-2.18(m,3H),2.02-1.88(m,2H),1.75-1.26(m,12H),1.20(s,3H).MS:(ES)m / z C 32 H 38 F3N4O4[M+H] + Calculated value: 599.3, actual value: 599.3.

[0249] Isomer 2: 1H NMR(400MHz,CD3OD)δ 8.14-8.00(m,1H),7.32-7.18(m,2H),7.16-7.08(m,2H),7.01-6.92(m,1H),6.70-6.60(m,1H),4.53-4.3 1(m,4H),3.15-3.05(m,3H),2.23(s,3H),1.85-1.69(m,4H),1.67-1.33(m,10H),1.20(s,3H).MS:(ES)m / z C 32 H 38 F3N4O4[M+H] + Calculated value: 599.3, actual value: 599.3.

[0250] Example 40: 5-(2,4-difluorophenyl)-N-(3-(2-((2-(4,6-dimethylpyrimidin-2-yl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-1,3,4-thiadiazole-2-carboxamide [ka] Step a: To a solution of tert-butyl 3-amino-3-(2-ethoxy-2-oxoethyl)azetidine-1-carboxylate (2.31 g, 8.96 mmol) and lithium 5-(2,4-difluorophenyl)-1,3,4-thiadiazole-2-carboxylate (2.22 g, 8.96 mmol) in 20 mL of DMF was added HATU (3.75 g, 9.86 mmol). The mixture was stirred at room temperature for 1 h and then quenched with H2O. The precipitated solid was filtered, rinsed with water, and dried in a vacuum oven to give tert-butyl 3-(5-(2,4-difluorophenyl)-1,3,4-thiadiazole-2-carboxamido)-3-(2-ethoxy-2-oxoethyl)azetidine-1-carboxylate. MS: (ES) m / z C 21 H 25 F2N4O5S[M+H] + The calculated value was 483.1, and the measured value was 483.2.

[0251] Step b: A mixture of tert-butyl 3-(5-(2,4-difluorophenyl)-1,3,4-thiadiazole-2-carboxamido)-3-(2-ethoxy-2-oxoethyl)azetidine-1-carboxylate (2.36 g, 4.89 mmol) in 4 M HCl in dioxane (20 mL) was stirred for 1 h. The contents were concentrated under reduced pressure to give ethyl acetate 2-(3-(5-(2,4-difluorophenyl)-1,3,4-thiadiazole-2-carboxamido)azetidin-3-yl). MS: (ES) m / z C 16 H 17 F2N4O3S[M+H] + Calculated value: 383.1, actual value: 383.1.

[0252] step c: A mixture of ethyl acetate 2-(3-(5-(2,4-difluorophenyl)-1,3,4-thiadiazole-2-carboxamido)azetidin-3-yl) (2.0 g, 5.23 mmol), pyridine (0.85 mL, 10.46 mmol) and 4-hydroxy-4-methylcyclohexan-1-one (1.0 g, 7.85 mmol) in 4:1 DCM / MeOH (50 mL) was stirred at room temperature for 1 h. To the contents was added NaBH(OAc)3 (2.22 g, 10.46 mmol). The reaction mixture was stirred at room temperature for an additional 1 h, then quenched with saturated NaHCO3(aq) and extracted with DCM. The combined organic layers were dried over MgSO4, filtered and concentrated in vacuo. The crude residue was purified by silica gel column chromatography to give two separated isomers of ethyl acetate 2-(3-(5-(2,4-difluorophenyl)-1,3,4-thiadiazole-2-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl). MS: (ES) m / z C 23 H 29 F2N4O4S[M+H] + Calculated value: 495.2, measured value: 495.2.

[0253] Step d: To a solution of ethyl acetate 2-(3-(5-(2,4-difluorophenyl)-1,3,4-thiadiazole-2-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl) (472 mg, 0.95 mmol) in 4:1 THF / HO (2 mL) was added LiOH (48 mg, 1.2 mmol). The mixture was stirred at room temperature overnight, then concentrated to dryness, acidified to pH 3-4, and extracted with a solution of CHCl3 / IPA (2:1). The organic layers were combined, dried, filtered, and concentrated to give 2-(3-(5-(2,4-difluorophenyl)-1,3,4-thiadiazole-2-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid. MS: (ES) m / z C 21 H 25 F2N4O4S[M+H] + Calculated value: 466.2, actual value: 466.2.

[0254] Step e: To a mixture of 2-(3-(5-(2,4-difluorophenyl)-1,3,4-thiadiazole-2-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (47 mg, 0.10 mmol) in DMF (2 mL) was added 2-(4,6-dimethylpyrimidin-2-yl)propan-2-amine (17 mg, 0.10 mmol), DIPEA (0.04 mL, 0.24 mmol) and HATU (42 mg, 0.11 mmol). The contents were stirred for 1 h and then purified by preparative HPLC to give the desired product, 5-(2,4-difluorophenyl)-N-(3-(2-((2-(4,6-dimethylpyrimidin-2-yl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-1,3,4-thiadiazole-2-carboxamide.

[0255] Isomer 1: 1H NMR(400MHz,CD3OD)δ 8.44(ddd,J=8.0,8.0,8.0Hz,1H),7.37-7.20(m,2H),7.05(s,1H),4.64-4.40(m,4H),3.43-3.22(m,1H), 3.13(d,J=9.3Hz,2H),2.40(s,6H),1.99(s,2H),1.75-1.35(m,6H),1.62(s,6H),1.21(s,3H).MS:(ES)m / z C 30 H 38 F2N7O3S[M+H] + Calculated value: 614.3, actual value: 614.0.

[0256] Isomer 2: 1 H NMR(400MHz,CD3OD)δ 8.50-8.40(m,1H),7.35-7.22(m,2H),7.05(s,1H),4.62-4.40(m,4H),3.39-3.22(m,3H), 2.39(s,6H),1.84-1.71(m,4H),1.61(s,6H),1.62-1.35(m,4H),1.21(s,3H).MS:(ES)m / z C 30 H 37 F2N7O3S[M+H] + Calculated value: 614.3, actual value: 614.0.

[0257] Example 41: N-(3-(2-((2-(3-chlorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-5-(2,4-difluorophenyl)thiazole-2-carboxamide [ka] Step a: To a solution containing (2,4-difluorophenyl)boronic acid (370 mg, 2.3 mmol) and ethyl 5-bromothiazole-2-carboxylate (500 mg, 2.1 mmol) in a 2:1 mixture of toluene / HO (4.2 mL / 2.1 mL) was added Pd(OAc) (48 mg, 0.21 mmol), Xantphos (98 mg, 0.21 mmol) and NMM (0.52 mL, 4.7 mmol). After 2 h, the mixture was extracted with EtOAc. The organic layers were combined, dried over sodium sulfate, filtered and concentrated. The crude material was purified by silica gel column chromatography to give ethyl 5-(2,4-difluorophenyl)thiazole-2-carboxylate. MS: (ES) m / z C 12 H 10 F2NO2S[M+H] + The calculated value was 270.0 and the measured value was 270.1.

[0258] Step b: To a solution of ethyl 5-(2,4-difluorophenyl)thiazole-2-carboxylate (488 mg, 1.8 mmol) in 5.4 mL of THF was added a solution of 1 M LiOH (5.4 mL). The reaction mixture was stirred at room temperature for 16 h and then quenched with 1 N HCl. The contents were filtered and the solid was collected and dried to give 5-(2,4-difluorophenyl)thiazole-2-carboxylic acid. MS: (ES) m / z C 10 H6F2NO2S[M+H] + Calculated values ​​are 242.0 and 241.9.

[0259] Step c: To a solution containing 5-(2,4-difluorophenyl)thiazole-2-carboxylic acid (450 mg, 1.9 mmol) and tert-butyl 3-amino-3-(2-ethoxy-2-oxoethyl)azetidine-1-carboxylate (440 mg, 1.7 mmol) in 6.8 mL of DMF was added DIPEA (0.59 mL, 3.4 mmol) followed by HATU (780 mg, 2.1 mmol). The contents were stirred at room temperature for 16 hours, then concentrated under reduced pressure and purified by silica gel column chromatography to give tert-butyl 3-(5-(2,4-difluorophenyl)thiazole-2-carboxamido)-3-(2-ethoxy-2-oxoethyl)azetidine-1-carboxylate. MS: (ES) m / z C 22 H 26 F2N3O5S[M+H] + Calculated value: 482.2, measured value: 482.2.

[0260] Step d: To a solution of tert-butyl 3-(5-(2,4-difluorophenyl)thiazole-2-carboxamido)-3-(2-ethoxy-2-oxoethyl)azetidine-1-carboxylate (820 mg, 1.7 mmol) in 3.6 mL of dioxane was added a solution of 4.0 M HCl in dioxane (3.6 mL, 14.2 mmol). The contents were stirred at room temperature for 16 h and then concentrated to give ethyl acetate 2-(3-(5-(2,4-difluorophenyl)thiazole-2-carboxamido)azetidin-3-yl). MS: (ES) m / z C 17 H 18 F2N3O3S[M+H] + The calculated value was 382.1, and the measured value was 382.2.

[0261] Step e: To a solution of ethyl acetate 2-(3-(5-(2,4-difluorophenyl)thiazole-2-carboxamido)azetidin-3-yl) (360 mg, 0.94 mmol) and 4-hydroxy-4-methylcyclohexan-1-one (165 mg, 1.3 mmol) in 3.4 mL of DCM was added pyridine (0.15 mL, 1.4 mmol). After stirring at room temperature for 10 min, NaBH(OAc) (360 mg, 1.7 mmol) was added and the contents were stirred for an additional 16 h. The reaction was quenched with H2O and extracted with EtOAc. The combined organic layers were dried over sodium sulfate, filtered and concentrated. The crude material was purified by silica gel column chromatography to give ethyl acetate 2-(3-(5-(2,4-difluorophenyl)thiazole-2-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl). MS: (ES) m / z C 24 H 30 F2N3O4S[M+H] + The calculated value was 494.2, and the measured value was 494.4.

[0262] Step f: To a solution of ethyl acetate 2-(3-(5-(2,4-difluorophenyl)thiazole-2-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl) (339 mg, 0.69 mmol) in 2 mL of THF was added a solution of 1 M LiOH (2 mL, 2.0 mmol). The contents were stirred at room temperature for 16 h, then quenched with 1 N HCl and concentrated to dryness to give 2-(3-(5-(2,4-difluorophenyl)thiazole-2-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid. MS: (ES) m / z C 22 H 26 F2N3O4S[M+H] + The calculated value was 466.2, and the measured value was 466.3.

[0263] Step g: To a solution of 2-(3-(5-(2,4-difluorophenyl)thiazole-2-carboxamido)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)acetic acid (160 mg, 0.35 mmol) and 2-(3-chlorophenyl)propan-2-amine (70 mg, 0.41 mmol) in 2 mL of DMF was added DIPEA (0.19 mL, 1.1 mmol) followed by HATU (245 mg, 0.65 mmol). The contents were stirred at room temperature for 4 h and then concentrated under reduced pressure. The crude material was purified by preparative HPLC to give two separated isomers of N-(3-(2-((2-(3-chlorophenyl)propan-2-yl)amino)-2-oxoethyl)-1-(4-hydroxy-4-methylcyclohexyl)azetidin-3-yl)-5-(2,4-difluorophenyl)thiazole-2-carboxamide.

[0264] Isomer 1: 1 H NMR(400MHz,CD3OD)δ 8.31(m,1H),7.88(ddd,J=8.1,8.1,8.1Hz,1H),7.38-7.05(m,6H),4.68-4.29(m,4H),3.13(m,2H),2.05-1.90(m, 2H),1.78-1.62(m,2H),1.59(s,3H),1.57(s,3H),1.55-1.44(m,3H),1.44-1.28(m,2H),1.20(s,3H).MS:(ES)m / z C 31 H 36 ClF2N4O3S[M+H] + Calculated value: 617.2, measured value: 617.4.

[0265] Isomer 2: 1H NMR (400MHz, CD3OD) δ 8.49(d,J=10.6Hz,1H),8.32(d,J=3.4Hz,1H),7.92-7.84(m,1H),7.30-7.27( m,1H),7.25-7.17(m,2H),7.17-7.10(m,4H),4.59-4.43(m,4H),4.37(d,J=11. 8Hz,1H),3.17-2.94(m,3H),1.85-1.70(m,3H),1.64-1.58(m,1H),1.57(s,3H) ),1.56(s,3H),1.55-1.49(m,1H),1.47-1.35(m,2H),1.20(s,3H).MS:(ES)m / z C 31 H 36 ClF2N4O3S[M+H] + Calculated value: 617.2, measured value: 617.4.

[0266] Biological Example 1 CXCR7 is fused in frame with a small enzyme donor fragment, ProLink, and co-expressed in CHO cells stably expressing a fusion protein of β-arrestin and a larger N-terminal deletion mutant of β-galactosidase (termed the enzyme acceptor or EA). Activation of CXCR7 stimulates the binding of β-arrestin to ProLink-tagged CXCR7, allowing complementation of the two enzyme fragments to form the active β-galactosidase enzyme. β-galactosidase enzyme activity is measured with a fluorogenic substrate.

[0267] CHO-CXCR7 cells (0.22×10 6 Cells were cultured in growth medium (Ham's F12 medium containing 10% fetal bovine serum (FBS)) and the transgene was maintained using hygromycin B (200 μg / mL) and G418 (250 μg / mL). The day before the assay, cells were detached from the culture dish using 0.25% trypsin-EDTA (Corning, catalog no. 25-053-CI) and plated in 96-well plates (2.2 × 10 5Cells / mL, 100 μL / well) were plated and incubated overnight at 37° C. with 5% CO2. On the day of the assay, cell medium was removed and 100 μL of assay buffer (PBS or FBS) was added to each well. 1 μL of compound serially diluted in DMSO was added to each well. 5 μL of human SDF-1a (Pepro Tech, Cat. No. 300-28A, at EC50 concentration previously determined on the same day) was subsequently added and mixed to induce CXCR7-mediated β-arrestin recruitment. Plates were incubated at 37° C. for 1.5 hours. Assay buffer was removed and 100 μL of substrate solution was added and the reaction was carried out at 37° C. for 30 minutes. Substrate solution was prepared by mixing 100 mL of phosphate buffer (1 M, Sigma, Cat. No. P3619-1GA), 100 mL of 10% Triton X 100 (Sigma, Cat. No. T8787), 5 mL of MgCl2 (1 M, Sigma, Cat. No. M1028), and 1.5 mL of β-mercaptoethanol (Gibco, Cat. No. 21985-023). 3-Carboxyumbelliferyl β-D-galactopyranoside (ThermoFisher Scientific, Cat. No. F2905) was added at 0.4 mM immediately before use. The reaction was subsequently stopped by adding 50 μL of stop solution (2.1% w / v Na2CO3) to each well. Fluorescence intensity was measured on a FlexStation3 microplate reader (Molecular Devices) using the following settings: excitation: 360 nm, emission: 465 nm, manual gain: 55. IC 50 Values ​​were calculated using three-parameter nonlinear regression in GraphPad Prism.

[0268] verification Compounds initially identified as being of interest by any of the above screening methods can be further tested to verify the apparent activity in vivo. Preferably, such studies are carried out in suitable animal models. The basic format of such methods involves administering lead compounds identified during initial screening to those that serve as human disease models and then determining whether the disease (e.g., cancer, myocardial infarction, wound healing, inflammatory disease, or other diseases associated with CXCR7) is in fact modulated and / or the disease or condition is ameliorated. The animal models used in validation studies are generally any type of mammal. Specific examples of suitable animals include, but are not limited to, primates, mice, rats, and zebrafish.

[0269] The compounds in the table below were prepared as described above. Where specific synthetic details are not provided, the compounds were prepared by the above method with minor variations and substitution of selected reagents. The respective activities are provided as follows: IC50<5nM (++++); 5nM to up to 100nM (+++); 100nM to up to 1000nM (++); and 1000nM to 20,000nM (+).

[0270] [Table 1]

[0271] [Table 2]

[0272] [Table 3]

[0273] [Table 4]

[0274]

Table 5

[0275]

Table 6

[0276]

Table 7

[0277]

Table 8

[0278]

Table 9

[0279]

Table 10

[0280]

Table 11

[0281]

Table 12

[0282]

Table 13

[0283]

Table 14

[0284]

Table 15

[0285]

Table 16

[0286]

Table 17

[0287]

Table 18

[0288]

Table 19

[0289]

Table 20

[0290]

Table 21

[0291]

Table 22

[0292]

Table 23

[0293]

Table 24

[0294]

Table 25

[0295]

Table 26

[0296]

Table 27

[0297]

Table 28

[0298]

Table 29

[0299]

Table 30

[0300]

Table 31

[0301]

Table 32

[0302]

Table 33

[0303]

Table 34

[0304]

Table 35

[0305]

Table 36

[0306]

Table 37

[0307]

Table 38

[0308]

Table 39

[0309]

Table 40

[0310]

Table 41

[0311]

Table 42

[0312]

Table 43

[0313]

Table 44

[0314]

Table 45

[0315]

Table 46

[0316]

Table 47

[0317]

Table 48

[0318]

Table 49

[0319]

Table 50

[0320]

Table 51

[0321]

Table 52

[0322]

Table 53

[0323]

Table 54

[0324]

Table 55

[0325]

Table 56

[0326]

Table 57

[0327]

Table 58

[0328]

Table 59

[0329]

Table 60

[0330]

Table 61

[0331]

Table 62

[0332]

Table 63

[0333]

Table 64

[0334]

Table 65

[0335]

Table 66

[0336]

Table 67

[0337]

Table 68

[0338]

Table 69

[0339]

Table 70

[0340]

Table 71

[0341]

Table 72

[0342]

Table 73

[0343]

Table 74

[0344]

Table 75

[0345]

Table 76

[0346]

Table 77

[0347]

Table 78

[0348]

Table 79

[0349]

Table 80

[0350]

Table 81

[0351]

Table 82

[0352]

Table 83

[0353]

Table 84

[0354]

Table 85

[0355]

Table 86

[0356]

Table 87

[0357]

Table 88

[0358] [Table 89]

[0359] [Table 90]

[0360] [Table 91]

[0361] Those skilled in the art will recognize from the description, figures and examples provided that modifications and variations can be made to the various embodiments of the present invention without departing from the scope of the present invention, which is defined by the following claims and their equivalents.

[0362] All patents, patent applications, publications and presentations cited herein are incorporated by reference in their entirety. Any conflict between any reference cited herein and the teachings of this specification shall be resolved in favor of the latter. Similarly, any conflict between an art-recognized definition of a term or phrase and a definition of a term or phrase provided herein shall be resolved in favor of the latter.

Claims

1. Formula (I): 【Chemistry 1】 (In the formula, HAr is a 5-membered heteroaryl ring; Ar １ is selected from the group consisting of phenyl, pyridyl, pyrimidinyl and pyrazinyl; Ar ２ are each independently a monocyclic or fused bicyclic aryl or heteroaryl; The subscript m is 0, 1, or 2; the subscript n is 0, 1, 2 or 3; the subscript p is 0, 1, 2 or 3; the subscript q is 0, 1, 2, 3 or 4; Each R １ are independently halogen, CN, C １～４ Alkyl, C １～４ Haloalkyl, -NR ａ R ｂ , -OR ａ , -CO ２ R ａ and -C(O)NR ａ R ｂ is a member selected from the group consisting of Each R ２ are independently halogen, CN, C １～４ Alkyl, C １～４ Haloalkyl, -NR ａ R ｂ , -OR ａ , -CO ２ R ａ and -C(O)NR ａ R ｂ is a member selected from the group consisting of Each R ３ is C １～４ Alkyl, C １～４ Haloalkyl, C １～４ Hydroxyalkyl, —CO ２ R ａ , -X-CO ２ R ａ , -C(O)NR ａ R ｂ and -X-C(O)NR ａ R ｂ is a member selected from the group consisting of R ４ａ and R ５ａ each independently represents H, C １～６ Alkyl, C １～６ Haloalkyl, C １～６ Hydroxyalkyl, -X-OR ａ , -CO ２ R ａ , -X-CO ２ R ａ , -X-NR ａ R ｂ , -C(O)NR ａ R ｂ and -X-C(O)NR ａ R ｂ is a member selected from the group consisting of R ４ and R ５ each independently represents H, C １～６ Alkyl, C １～６ Haloalkyl, C １～６ Hydroxyalkyl, -X-OR ａ , -CO ２ R ａ , -X-CO ２ R ａ , -X-NR ａ R ｂ , -C(O)NR ａ R ｂ and -X-C(O)NR ａ R ｂ or R ４ and R ５ are combined to form a 3-5 membered ring having 0 or 1 heteroatom ring vertex selected from O, S, or N, said 3-5 membered ring being unsubstituted or substituted with halogen, CN, C １～４ Alkyl, C １～４ Haloalkyl, C １～４ Alkoxy and C １～４ substituted with 1 to 4 substituents independently selected from the group consisting of haloalkoxy; Each R ６ are independently halogen, CN, -X-CN, C １～４ Alkyl, C １～４ Haloalkyl, C ３～６ Cycloalkyl, C １～４ Hydroxyalkyl, -OR ａ , -CO ２ R ａ , -X-CO ２ R ａ , -NR ａ R ｂ , -X-NR ａ R ｂ , -C(O)NR ａ R ｂ and -X-C(O)NR ａ R ｂ is a member selected from the group consisting of R ７ is C １～８ Alkyl, C ３～８ Hydroxyalkyl, C １～４ Alkoxy-C ２～４ Alkyl, -C(O)NH-C １～８ Alkyl, —C(O)—C １～８ Alkyl, -S(O) ２ -C １～８ Alkyl, C ３～８ Cycloalkyl, -X-C ３～８ Cycloalkyl, C ６～９ Spirocycloalkyl, -X-C ６～９ is a member selected from the group consisting of spirocycloalkyl, 4- to 7-membered heterocycloalkyl, —X-4- to 7-membered heterocycloalkyl, 7- to 11-membered spiroheterocycloalkyl, and —X-7- to 11-membered spiroheterocycloalkyl; ７ is substituted with 0 to 4 substituents independently selected from the group consisting of hydroxy, methyl, ethyl, hydroxymethyl, fluoro, chloro, methoxy, ethoxy, and cyclopropyl; Each R ａ and R ｂ are independently H, C １～４ Alkyl, C １～４ Haloalkyl, C ３～６ Cycloalkyl and C ３～６ Cycloalkyl-C １～４ is selected from the group consisting of alkyl, Each X is C １～４ is an alkylene linking group, and any of the methylene moieties of X is unsubstituted or substituted with one or two methyl groups. or a pharma- ceutically acceptable salt, hydrate, N-oxide, isotopically or enantiomerically enriched version, or rotamer thereof.

2. 2. The compound of claim 1, wherein HAr is selected from the group consisting of isoxazole, isothiazole, imidazole, pyrazole, thiazole, 1,2,4-oxadiazole, 1,3,4-oxadiazole, 1,2,4-thiadiazole, 1,3,4-thiadiazole, 1,2,3-triazole and 1,2,4-triazole.

3. 2. The compound of claim 1, wherein HAr is selected from the group consisting of isoxazole and thiadiazole.

4. Ar １ The compound of claim 1 , wherein is phenyl.

5. the subscript q is 1, 2, or 3, and each R １ are independently halogen, CN, C １～４ Alkyl and C １～４ 5. The compound of claim 4, which is a member selected from the group consisting of haloalkyl.

6. Ar ２ 2. The compound of claim 1, wherein is selected from the group consisting of pyridyl, pyrimidinyl, pyrazinyl, phenyl, indolyl, thiazolyl, pyrazolyl, indazolyl, and pyrrolopyridinyl.

7. Ar ２ The compound of claim 6, wherein is selected from the group consisting of pyrimidinyl, pyridyl and phenyl.

8. Ar ２ is selected from the group consisting of 2-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 2-thiazolyl, 4-pyrazolyl, phenyl, and indolyl, and each R ６ are independently halogen, CN, C １～４ Alkyl, C １～４ Haloalkyl, C ３～６ Cycloalkyl and C １～４ 2. The compound of claim 1, selected from the group consisting of alkoxy.

9. -Ar ２ - (R ６ ) ｐ teeth, 【Chemistry 2】 2. The compound of claim 1 selected from the group consisting of:

10. R ７ teeth, 【Chemistry 3】 2. The compound of claim 1 selected from the group consisting of:

11. The compound of claim 1 , wherein the subscript m is 0.

12. The compound of claim 1 , wherein the subscript n is 0.

13. 2. The compound of claim 1, wherein the subscript p is 0, 1 or 2.

14. The compound of claim 1 , wherein the subscript q is 1 or 2.

15. Formula (Ia): 【Chemistry 4】 2. The compound of claim 1 having the formula:

16. 16. The compound of claim 15, wherein HAr is selected from the group consisting of isoxazole and thiadiazole.

17. Formula (Ia1): 【Chemistry 5】 16. The compound of claim 15, having the formula:

18. Formula (Ia2): 【Chemistry 6】 18. The compound of claim 17, having the formula:

19. 20. The compound of claim 18, wherein HAr is selected from the group consisting of isoxazole and thiadiazole.

20. Formula (Ib), (Ic) or (Id): 【Chemistry 7】 2. The compound of claim 1 having the formula:

21. Formula (Ib1), (Ic1) or (Id1): 【Chemistry 8】 21. The compound of claim 20, having the formula:

22. 22. The compound of claim 21, wherein HAr is selected from the group consisting of isoxazole and thiadiazole.

23. R ７ is C １～８ Alkyl, C ３～８ Hydroxyalkyl, C １～４ Alkoxy-C ２～４ Alkyl, C ３～８ Cycloalkyl, C ６～９ is a member selected from the group consisting of spirocycloalkyl, 4- to 7-membered heterocycloalkyl, and 7- to 11-membered spiroheterocycloalkyl; ７ is substituted with 0 to 4 substituents independently selected from the group consisting of hydroxy, methyl, ethyl, hydroxymethyl, fluoro, chloro, methoxy, ethoxy, and cyclopropyl.

24. R ７ is -X-C ３～８ Cycloalkyl, -X-C ６～９ is a member selected from the group consisting of spirocycloalkyl, -X-4 to 7 membered heterocycloalkyl, and -X-7 to 11 membered spiroheterocycloalkyl, ７ is substituted with 0 to 4 substituents independently selected from the group consisting of hydroxy, methyl, ethyl, hydroxymethyl, fluoro, chloro, methoxy, ethoxy, and cyclopropyl.

25. R ７ is selected from the group consisting of cyclohexyl, cyclopentyl, piperidinyl, tetrahydropyranyl, and tetrahydrofuranyl, each of which is substituted with 0 to 2 substituents independently selected from the group consisting of hydroxy, methyl, ethyl, hydroxymethyl, fluoro, chloro, methoxy, and ethoxy.

26. 2. A compound according to claim 1 selected from the compounds of the examples.

27. 2. The compound of claim 1 selected from those in Table 1.

28. 13. A pharmaceutical composition comprising a compound of claim 1 and a pharma- ceutically acceptable excipient.

29. 29. The pharmaceutical composition of claim 28, wherein the compound is a compound of Table 1.

30. 1. A pharmaceutical composition for use in a method of treating a disease or disorder in a mammal, comprising: The method comprises administering to the subject a therapeutically effective amount of a compound of claim 1 for a period of time sufficient to treat the disease or disorder; and 13. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to claim 1.

31. 31. The pharmaceutical composition of claim 30, wherein the compound is a compound of Table 1.

32. 31. The pharmaceutical composition of claim 30, wherein the disease or disorder is selected from the group consisting of cancer, inflammation and neurological or progenitor / stem cell disorders.

33. 11. A method of inhibiting binding of the chemokine I-TAC or SDF-1 to a CXCR7 receptor, comprising contacting a cell expressing the CXCR7 receptor with a compound of claim 1 for a period of time sufficient to inhibit the binding of the chemokine to the CXCR7 receptor.

34. 34. The method of claim 33, wherein the compound is a compound of Table 1.

35. 1. A method for imaging a tumor, organ or tissue, comprising: (a) administering to a subject in need of said imaging a compound according to any one of claims 1 to 15 in a radiolabeled or detectable form; (b) detecting the compound and determining where the compound is concentrated in the subject; The method includes:

36. 36. The method of claim 35, wherein the compound is radiolabeled.

37. 1. A method for detecting elevated levels of CXCR7 in a sample, comprising: (a) contacting a sample suspected of having elevated levels of CXCR7 with a compound according to any one of claims 1 to 15 in radiolabelled or detectable form; (b) determining the level of a compound bound to CXCR7 present in the sample to determine the level of CXCR7 present in the sample; (c) comparing the level determined in step (b) to a control sample to determine whether an elevated level of CXCR7 is present in the sample; The method includes:

38. 38. The method of claim 37, wherein the compound is radiolabeled.

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