Modulators of chemokine receptors

Abstract

Compounds having the formula (A) are provided as chemotactic factor inhibitors:

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Application No. 62 / 257,389, filed November 19, 2015, and U.S. Provisional Application No. 62 / 277,711, filed January 12, 2016, pursuant to 35 U.S. SC §119(e), the entirety of which is incorporated herein by reference.

[0003] Claims of ownership for inventions made under federally funded research and development

[0004] not applicable

[0005] References include "sequence lists," tables, or computer program listing attachments submitted on CD-ROM.

[0006] not applicable Background of the Invention

[0008] Chemokines are chemokines released by various cells to attract macrophages, lymphocytes, eosinophils, basophils, and neutrophils to sites of inflammation (reviewed in Schall, Cytokine, 3:165-183 (1991), Schall et al., Curr Opin. Immunol. 6:865-873 (1994), and Murphy, Rev. Immun. 12:593-633 (1994)). In addition to stimulating chemotaxis, chemokines can selectively induce other changes in responding cells, including changes in cell shape and the release of intracellular free calcium ions ([Ca2+]). 2+ The transient increase in inflammatory concentrations, granule exocytosis, integrin upregulation, formation of bioactive lipids (such as leukotrienes), and respiratory burst are all related to leukocyte activation. Therefore, chemokines are early triggers of inflammatory responses, causing the release of inflammatory mediators, chemotaxis, and extravasation to sites of infection or inflammation.

[0009] There are two main classes of chemokines, CXC (α) and CC (β), depending on whether the first two cysteine ​​residues are separated by a single amino acid (CXC) or adjacent to each other (CC). α-chemokines such as CXCL1 (GROα) and CXCL8 (interleukin-8, IL-8) are primarily chemotactic towards neutrophils, while β-chemokines such as CCL5 (RANTES) and CCL20 (LARC, MIP-3α) are chemotactic towards T cells, B cells, macrophages, eosinophils, and basophils (Deng et al., Nature, 381:661-666 (1996)). Chemokines bind to specific cell surface receptors belonging to the G protein-coupled seven-transmembrane domain protein family (reviewed in Horuk, Trends Pharm.Sci., 15:159-165 (1994)), which are referred to as "chemokine receptors".

[0010] Upon binding to their homologous ligands, chemokine receptors transpose intracellular signals via associated trimeric G proteins, leading to a rapid increase in intracellular calcium concentration. At least 11 human chemokine receptors bind to or respond to β-chemokines, and at least 7 bind α-chemokines. Furthermore, CX3CR1 (fractalkine receptor) can bind fractal chemokines, distinguished by a series of three amino acids between the first two cysteine ​​residues. Chemokine receptors are important mediators of inflammatory and immune regulatory disorders and diseases, including asthma and allergic diseases, as well as autoimmune diseases such as rheumatoid arthritis and atherosclerosis.

[0011] The chemokine receptor CCR6 is known to be expressed by memory (but not primary) CD4 T cells, IL-17-secreting αβ T cells, IL-17-secreting γδ T cells, regulatory T cells, B cells, and dendritic cells. Its only known ligand is CCL20 (MIP-3α, LARC), which shows strong binding to others. It is expressed on approximately 30–60% of adult peripheral blood effector / memory CD4+ T cells. CCR6 is involved in leukocyte homing to inflamed tissues, particularly the skin, lungs, and intestines; and is co-expressed on a subset of T cells with a skin-homing phenotype (i.e., T cells expressing skin lymphocyte antigen (CLA) and CCR4). Therefore, CCR6 may be an important player in leukocyte involvement in skin lesions.

[0012] CCR6 expression is associated with psoriasis. In humans, most skin-homing CD4 T cells in peripheral blood that express IL17 express CCR6 (Homey et al., JI, 2000). In several inflammatory diseases, IL17-secreting cells act as central mediators. T cells, such as γδ T cells and TH17 T cells, produce IL17 upon activation. The pathogenic effects of IL17 are associated with human diseases such as rheumatoid arthritis (Patel DD et al., Ann Rheum Dis 2013), multiple sclerosis (Zepp J, Wu L, and X Li, Trends Immunol 2011), and psoriasis (Martin DA et al., J. Invest Dermatol 2012). Evidence for a strong link between IL17 and psoriasis includes genome-wide association studies that have shown a close association between psoriasis and upstream genes (IL-23) or downstream genes (NFb) of the IL17 signaling pathway, as well as a strong correlation with the efficacy of targeting IL17 in a clinical setting (Martin DA et al., J. Invest Dermat. 2012; Papp et al., NEJM, 2012; Papp et al., NEJM, 2012). In addition to enhanced CCL20-mediated chemotaxis, CCR6+ T cells isolated from psoriasis patients preferentially secrete IL-17A, IL22, and TNFα compared to healthy controls (Kagami et al., J. Invest. Dermatol., 2010). Finally, CCL20 mRNA expression is upregulated in focal psoriatic skin samples (Homey et al., JI, 2000; Dieu-Nosjean et al., JEM, 2000). In mice, CCR6 knockout mice are protected from IL-23-driven psoriasis (Hedrick MN et al. JCI, 2009). Therefore, substantial evidence from mice and humans suggests the protective role of CCR6 blockade in psoriasis and psoriasis-like models.

[0013] CCR6 is also expressed by dendritic cells during key stages of their development and is important for their migration through tissues (Sozzani et al., J Leuk Biol, 66:1, 1999). Dendritic cells are responsible for presenting antigens to T cells within lymph nodes; therefore, inhibiting dendritic cell transport can suppress T cell-mediated inflammatory responses (Banchereau and Steinman, Nature, 392:245, 1998).

[0014] CCR6 is expressed by B cells, and CCR6-mediated B cell migration has recently been shown to be essential for B cell memory responses to soluble antigens (Elgueta et al., J Immunol, 194:505, 2015). Therefore, inhibiting this B cell migration by blocking CCR6 may potentially suppress B cell-mediated (and therefore antibody-mediated) inflammatory responses in diseases such as lupus, rheumatoid arthritis, and pemphigus.

[0015] CCR6 is commonly expressed by colorectal cancer (CRC) cells. High expression of this receptor is associated with poor outcomes in CRC patients, and it has been proposed that CCR6 itself contributes to the migration of CRC cells that lead to metastasis (Liu J. et al., Plosone 20149(6):e101137).

[0016] Chemokines that bind to the receptor CXCR2 alone promote the accumulation and activation of neutrophils. These chemokines are involved in a wide range of acute and chronic inflammatory diseases, such as psoriasis, rheumatoid arthritis, radiation-induced fibrotic lung disease, autoimmune bullous skin disease (AIBD), chronic obstructive pulmonary disease (COPD), and ozone-induced airway inflammation (see Baggiolini et al., Febs Lett. 307: 97 (1992); Miller et al., Crit. Rev. Immunol. 12: 17 (1992); Oppenheim et al., annu. Rev. Immunol. 9: 617 (1991); Seitz et al., J. Clin. Invest. 87: 463 (1991); Miller et al., Ann. Rev. Respir. Dis. 146: 427 (1992); and Donnely et al., Lancet 341: 643 (1993), Fox & Haston, Radiation Oncology, 85:215 (2013), Hirose et al., J. Genet. Syndr. Genet. Ther. S3:005 (2013), Miller et al., Eur. J. Drug Metab. Pharmacokinet. 39:173 (2014), Lazaar et al., Br. J. Clin. Pharmacol., 72:282 (2011)).

[0017] Besides inflammatory diseases, several CXCR2 ligand chemokines, including CXCL1, CXCL2, CXCL3, and CXCL5, are also involved in inducing tumor angiogenesis (Strieter et al. JBC 270:27348-27357 (1995)). During ischemic stroke, some CXCR2 ligand chemokines are aggravating agents (Connell et al. Neurosci. Lett., 15:30111 (2015). Their angiogenic activity may be due to the activation of CXCR2 expressed on the surface of vascular endothelial cells (ECs) of peripheral blood vessels by these chemokines.

[0018] Many types of tumors are known to produce CXCR2 ligand chemokines. The production of these chemokines is associated with a more aggressive phenotype (Inoue et al., Clin Cancer Res 6: 2104-2119 (2000)) and poor prognosis (Yoneda et al., J Nat Cancer Inst 90: 447-454 (1998)). Since these chemokines are potent chemokines for EC chemotaxis, they may play a role in inducing endothelial cells to migrate to their production sites within the tumor. This could be a key step in inducing tumor angiogenesis. CXCR2 inhibitors will suppress the angiogenic activity of ELR-CXC chemokines and thus block tumor growth. The antitumor activity of antibodies against CXCL8 (Arenberg et al., J Clin Invest 97: 2792-2802 (1996)), ENA-78 (Arenberg et al., J Clin Invest 102: 465-72 (1998)) and CXCL1 (Haghnegahdar et al., J. Leukoc Biology 67: 53-62 (2000)) has been confirmed.

[0019] Many tumor cells express CXCR2, and these cells can stimulate their own growth by secreting the ELR-CXC chemokine. Therefore, in addition to reducing angiogenesis within tumors, CXCR2 inhibitors can directly inhibit tumor cell growth.

[0020] CXCR2 is typically expressed by myeloid-derived suppressor cells (MDSCs) in the tumor microenvironment. MDSCs are involved in suppressing the tumor immune response, and the migration of CXCR2 ligand chemokines by MDSCs is most likely responsible for attracting these cells to the tumor (Marvel and Gabrilovich, J. Clin. Invest. 13:1 (2015) and Mackall et al., Sci. Trans. Med. 6:237 (2014)). Therefore, CXCR2 inhibitors can reverse the suppressive process and thereby enable immune cells to reject tumors more effectively. In fact, blocking the activation of CXC-chemokine receptors has been shown to be used in combination with checkpoint inhibitors to suppress tumor growth, suggesting that CXCR2 blockade can also be used in combination with other antitumor therapies (including but not limited to vaccines or conventional cytotoxic chemotherapy) to enhance tumor rejection (see Highfill et al., Science Translational Medicine, 6:237 (2014)).

[0021] The activities of CCR6 and CXCR2 are each associated with adverse outcomes in certain cancer types, including CRC, but they may exert their effects through different potential complementary mechanisms (Nandi et al., PLoS One, 9: e97566, 2014; Liu et al., PLoS One, 9: e101137, 2014; Chelvappa, Int J Colorectal Dis, 29: 1181, 2014; Zhang, Biomed Pharmacother. 69: 242, 2014; Lee et al., Int J Cancer, 135: 232, 2014; Wang and DuBois, Oncoimmunology, 29: e28581, 2014; Wu et al., Int J Clin Exp Med, 8: 5883, 2015).

[0022] Given the clinical importance of CCR6 and CXCR2, identifying compounds that regulate the function of one or both of these receptors represents an attractive avenue for developing novel therapeutics. This article provides information on these compounds and their methods of use. Summary of the Invention

[0023] This article describes compounds having formula (A):

[0024]

[0025] Where R 3 R 4 R 5a R 5b R 6a R6b R 7 B and the subscript n have the meanings provided in the specific description below. This compound can be used to treat diseases or conditions that are at least partially regulated by CCR6, and also to treat diseases or conditions that are at least partially regulated by CXCR2.

[0026] Pharmaceutical compositions of the compound of formula (A) are also provided.

[0027] This disclosure further provides a method for preparing the synthetic compound (A), and a selection of intermediates for the preparation. Attached Figure Description

[0028] Figure 1A-1AJ The structure and biological activities of the compounds described in this article are provided.

[0029] Figure 2 Compound 1.023 was shown in an IL-23-induced ear swelling model.

[0030] Figure 3 PASI scores are provided in mice treated with compound 1.129 in an imiquimod-induced psoriasis model.

[0031] Figure 4 The study demonstrated that mice treated with compound 1.129 had thickness, erythema, and desquamation scores ≥3 compared to mice treated with the vector in an imiquimod-induced psoriasis model. Invention Details

[0033] Before further describing the invention, it should be understood that the invention is not limited to the specific embodiments set forth herein, and it should also be understood that the terminology used herein is for the purpose of describing specific embodiments only and is not intended to be limiting.

[0034] Where a range of values ​​is provided, it should be understood that the invention includes every intermediate value between the upper and lower limits of that range, as well as any other specified or intermediate value within that range, accurate to one-tenth of the lower limit unit unless the context explicitly specifies otherwise. The upper and lower limits of these smaller ranges may be independently included within the smaller range and also within the invention, subject to any specifically excluded limitations within the range. Where the range includes one or both limitations, the range excluding any one or both of those included limitations is also included in the invention. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0035] It must be noted that, unless the context clearly indicates otherwise, the singular forms “a,” “an,” and “the” as used herein and in the appended claims include plural indicators. It should be further noted that claims may be drafted to exclude any optional elements. Therefore, this statement is intended to serve as a preliminary basis for the use of exclusive terms such as “merely,” “only,” or the use of a “negative” limitation.

[0036] The publications discussed herein are provided only because they were published prior to the filing date of this application. Furthermore, the publication dates provided may differ from the actual publication dates and may require independent verification.

[0037] Overview

[0038] This invention derives from the discovery of compounds of formulas (A), (A1), (A2), (I), and (Ia1) as potent antagonists of the CCR6 receptor and / or CXCR2 receptor. These compounds possess in vivo anti-inflammatory activity and exhibit excellent pharmacokinetic properties. Therefore, the compounds provided herein can be used in pharmaceutical compositions, methods for treating CCR6-mediated and / or CXCR2-mediated diseases, and as controls in assays for identifying CCR6 and / or CXCR2 antagonists.

[0039] Abbreviations and Definitions

[0040] Unless otherwise stated, the following terms are intended to have the meanings described below. Other terms are defined elsewhere throughout the specification.

[0041] Unless otherwise stated, the term "alkyl" on its own or as part of another substituent refers to having a specified number of carbon atoms (i.e., C40, C50, C6 ... 1-8 Alkyl groups are straight-chain or branched hydrocarbon groups (representing 1-8 carbons). Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, etc.

[0042] The term "cycloalkyl" refers to a ring with a specified number of ring atoms (e.g., C10, C20, C30, C40, C50, C60, C7 ... 3-6 Cycloalkyl) and a hydrocarbon ring that is fully saturated or has no more than one double bond between the ring vertices. “Cycloalkyl” also refers to bicyclic and polycyclic hydrocarbon rings, such as bicyclic [2.2.1]heptane, bicyclic [2.2.2]octane, etc.

[0043] The term "cycloheteroalkyl" refers to a cycloalkyl ring having a specified number of ring vertices (or members) and 1-5 heteroatoms selected from N, O, and S, wherein the heteroatoms substitute for 1-5 carbon vertices, and wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom is optionally quaternized. Cycloheteroalkyl groups can be monocyclic, bicyclic, or polycyclic. Non-limiting examples of cycloheteroalkyl groups include pyrrolidine, imidazoline, pyrazolidine, butyrolactam, valeron, imidazolidinone, hydantoin, dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S, S-oxide, piperazine, pyran, pyridone, 3-pyrrolidine, thiaran, pyranone, tetrahydrofuran, tetrahydrothiophene, quinine ring, etc. Cycloheteroalkyl groups can be attached to the rest of the molecule via a ring carbon or heteroatom.

[0044] As used herein, in any chemical structure described herein, a wavy line running across a single, double, or triple bond... This indicates the point of connection between the single, double, or triple bond and the rest of the molecule. Furthermore, a bond extending to the center of the ring (e.g., a benzene ring) signifies a connection at any available ring vertices. Those skilled in the art will understand that multiple substituents shown as being attached to the ring will occupy ring vertices that provide stability to the compound and are otherwise spatially compatible. For divalent components, this designation signifies including either orientation (forward or reverse). For example, the group “-C(O)NH” signifies including a connection of either orientation: -C(O)NH- or –NHC(O)-, and similarly, “-O-CH2CH2-” is intended to include both -O-CH2CH2- and -CH2CH2-O-.

[0045] The terms "alkoxy," "alkylamino," and "alkathio" (or thioalkoxy) are used in their conventional sense to refer to alkyl groups attached to the remainder of a molecule via an oxygen, amino, or sulfur atom, respectively. Furthermore, for dialkylamino groups, the alkyl moiety can be the same or different, and can combine with the nitrogen atom attached to each alkyl group to form a 3-7 membered ring. Therefore, dialkylamino or -NR a R b The groups shown represent piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl, etc.

[0046] Unless otherwise stated, the terms "halogenated" or "halogen" themselves, or as part of another substituent, refer to a fluorine, chlorine, bromine, or iodine atom. Furthermore, terms such as "halogenated alkyl" indicate the inclusion of monohalogenated or polyhalogenated alkyl groups. For example, the term "C 1-4 "Halogenated alkyl" indicates that it includes trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, etc.

[0047] Unless otherwise stated, the term "aryl" refers to a polyunsaturated (usually aromatic) hydrocarbon group, which can be a monocyclic or fused together or covalently linked polycyclic (up to three rings). Non-limiting examples of aryl groups include phenyl, naphthyl, and biphenyl.

[0048] The term "heteroaryl" refers to an aryl group (or ring) containing 1-5 heteroatoms selected from N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and one or more nitrogen atoms are optionally quaternized. Heteroaryl groups can be attached to the rest of the molecule via heteroatoms. Non-limiting examples of heteroaryl groups include pyridinyl, pyrazinyl, pyrazinyl, pyrimidinyl, triazinyl, quinolinyl, quinoxalinyl, quinazolinyl, terpineyl, phthalazinyl, benzotriazinyl, purine, benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzoisoxazolyl, isobenzofuryl, isoindoleyl, and indoleyl (a nitrogen-containing group). (indolizinyl), benzotriazinyl, thienopyridyl, thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridine, benzothiazolyl, benzofuranyl, benzothiaphenyl, indolyl, quinolinyl, isoquinolinyl, isothiazolyl, pyrazolyl, indazoleyl, pteridinyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrroleyl, thiazolyl, furanyl, thiopheneyl, etc. The substituents of the heteroaryl ring may be selected from the acceptable substituents described below.

[0049] In some embodiments, the terms mentioned above (e.g., "alkyl", "aryl", and "heteroaryl") may be optionally substituted. Substituents for each type of group are provided below.

[0050] Alkyl groups (including those commonly referred to as alkylene, alkenyl, ynyl, and cycloalkyl) may optionally be substituents selected from a variety of groups from the group consisting of: halogen, -OR', -NR'R”, -SR', -SIR'R”R”', -OC(O)R', -C(O)R', -CO2R', -CONR'R”, -OC(O)NR'R”, -NR”C(O)R', -NR'-C(O)NR”R”', -NR” C(O)₂R', -NH-C(NH₂)=NH, -NR'C(NH₂)=NH, -NH-C(NH₂)=NR', -S(O)R', -S(O)₂R', -S(O)₂NR'R”, -NR'S(O)₂R”, -CN, and -NO₂, in quantities ranging from zero to (2m'+1), where m' is the total number of carbon atoms in this group. R', R”, and R”’ each independently represent hydrogen, unsubstituted C, and unsubstituted C. 1-8 Alkyl, unsubstituted aryl, aryl substituted with 1-3 halogens, unsubstituted C 1-8 Alkyl, C 1-8 Alkoxy or C 1-8Thioalkoxy or unsubstituted aryl-C 1-4 Alkyl groups. When R' and R” are attached to the same nitrogen atom, they can combine with the nitrogen atom to form 3-, 4-, 5-, 6-, or 7-membered rings. For example, -NR'R” refers to a ring that includes 1-pyrrolidinyl and 4-morpholinyl.

[0051] Similarly, the optional substituents for aryl and heteroaryl groups are diverse and are typically 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”', -NH -C(NH2)=NH, -NR'C(NH2)=NH, -NH-C(NH2)=NR', -S(O)R', -S(O)2R', -S(O)2NR'R”, -NR'S(O)2R”, -N3, perfluoro(C1-C4)alkoxy and perfluoro(C1-C4)alkyl, in numbers ranging from zero to the total number of open valences on the aromatic ring system; wherein R', R” and R”' are independently selected from hydrogen, C 1-8 Alkyl, C 1-8 Halogenated, C 3-6 cycloalkyl, C 2-8 alkenyl and C 2-8 Alkyne group. Other suitable substituents include each of the above aryl substituents attached to the ring atom via an alkylene chain of 1-4 carbon atoms.

[0052] The two substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be of the formula -TC(O)-(CH2). q The -U- group is substituted, where T and U are independently -NH-, -O-, -CH2-, or a single bond, and q is an integer from 0 to 2. Alternatively, the two substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced by the formula -A-(CH2). r -B-, where A and B are independently -CH2-, -O-, -NH-, -S-, -S(O)-, -S(O)2-, -S(O)2NR'- or single bonds, and r is an integer from 1 to 3. One single bond in the resulting new ring may optionally be replaced by a double bond. Alternatively, two substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced by the formula -(CH2). s -X-(CH2) t The substituents are substituted, where s and t are independent 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 C.1-6 alkyl.

[0053] As used in this article, the term “heteroatoms” is intended to include oxygen (O), nitrogen (N), sulfur (S), and silicon (Si).

[0054] When the variable (e.g., R) 1 or R a When a substituent appears more than once in any compound, its definition for each occurrence is independent of the definitions in other cases. Furthermore, combinations of substituents and / or variables are only permitted if such combinations produce stable compounds.

[0055] The term "pharmaceutically acceptable salt" is intended to include salts of active compounds prepared with relatively non-toxic acids or bases, depending on the specific substituents of the compounds described herein. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds in their neutral form with a sufficient amount of the desired base (pure or in a suitable inert solvent). Examples of salts derived from pharmaceutically acceptable inorganic bases include aluminum, ammonium, calcium, copper, iron, ferrous, lithium, magnesium, manganese, manganese, potassium, sodium, zinc, etc. Salts derived from pharmaceutically acceptable organic bases include salts of primary, secondary, and tertiary amines, including substituted amines, cyclic amines, naturally occurring amines, etc., 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, methylglucosamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds in neutral form with a sufficient amount of the desired acid (pure or in a suitable inert solvent). Pharmaceutically acceptable examples of acid addition salts include those derived from inorganic acids, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrocarbonic acid, phosphoric acid, monohydrophosphoric acid, dihydrophosphoric acid, sulfuric acid, monohydrosulfuric acid, hydroiodic acid, or phosphorous acid, etc.; and salts derived from relatively non-toxic organic acids, such as acetic acid, propionic acid, isobutyric acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, etc. Also included are salts of amino acids, such as arginine salts, and salts of organic acids, such as glucuronic acid or galactunoric acid (see, for example, Berge, SM, et al., “Pharmaceutical Salts”, Journal of Pharmaceutical Science, 1977, 66, 1-19). Some specific compounds of the present invention contain both basic and acidic functional groups, thereby enabling the compounds to be converted into basic addition salts or acid addition salts.

[0056] The neutral form of a compound can be regenerated by contacting the salt with a base or acid and separating the parent compound in a conventional manner. The parent form of the compound differs from the various salt forms in some physical properties (e.g., solubility in polar solvents), but otherwise, for the purposes of this invention, those salts are equivalent to the parent form of the compound.

[0057] In addition to salt forms, the present invention provides compounds in prodrug form. The prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of the present invention. Furthermore, prodrugs can be converted into the compounds of the present invention in an in vitro environment by chemical or biochemical methods. For example, when placed in a transdermal patch reservoir containing suitable enzymes or chemical reagents, the prodrug can be slowly converted into the compounds of the present invention.

[0058] Some compounds of the present invention may exist in both solvated and aqueous forms, including aqueous forms. The solvated forms are generally equivalent to the aqueous forms and should be included within the scope of the present invention. Some compounds of the present invention may exist in polymorphic or amorphous forms. For the purposes contemplated by the present invention, all physical forms are generally equivalent and should be included within the scope of the present invention.

[0059] Certain compounds of the present invention possess asymmetric carbon atoms (optical centers) or double bonds; racemates, diastereomers, geometric isomers, regioisomers, and individual isomers (e.g., isolated enantiomers) should all be included within the scope of the present invention. When a stereochemical description is shown, it means a compound in which one isomer is present and which is substantially free of the other isomer. “Substantially free” of the other isomer means a ratio of at least 80 / 20 between the two isomers, more preferably 90 / 10 or 95 / 5 or more. In some embodiments, one of the isomers will be present in an amount of at least 99%.

[0060] The compounds of the present invention may also contain, at one or more atoms constituting such compounds, atomic isotopes in non-natural proportions. A non-natural proportion of an isotope can be defined as ranging from the amount of the atom in question that is naturally found to 100% of that atom. For example, the compounds may be doped with radioactive isotopes, such as tritium (…). 3 H), Iodine-125 ( 125 I) or carbon-14 ( 14 C), or non-radioactive isotopes, such as deuterium (C). 2 H) or carbon-13 ( 13C). In addition to the uses described in this application, such isotopic variants may provide additional uses. For example, isotopic variants of the compounds of the present invention may have additional uses, including but not limited to, as diagnostic and / or imaging agents, or as cytotoxic / radiotoxic therapeutic agents. Furthermore, isotopic variants of the compounds of the present invention may have altered pharmacokinetic and pharmacodynamic characteristics, thereby contributing to increased safety, tolerability, or efficacy during treatment. All isotopic variants of the compounds of the present invention, regardless of radioactivity, should be included within the scope of this invention.

[0061] The terms “patient” or “subject” are used interchangeably to refer to humans or non-human animals (e.g., mammals).

[0062] When applied to subjects, cells, tissues, organs, or biological fluids, the terms "administration," "giving," etc., refer to the contact between a CCR6 and / or CXCR2 antagonist, a pharmaceutical composition comprising it, or a diagnostic agent and a subject, cell, tissue, organ, or biological fluid. In the case of cells, administration includes contacting the reagent with cells (e.g., in vitro or ex vivo) and contacting the reagent with a fluid, wherein the fluid contacts the cells.

[0063] The terms "treat," "treating," and "treatment" refer to a process of action initiated after a disease, condition, or symptom, or its symptoms, has been diagnosed or observed (e.g., administration of an antagonist of CCR6 and / or CXCR2 or a pharmaceutical composition containing such antagonists) to temporarily or permanently eliminate, alleviate, suppress, slow down, or improve the disease, condition, or symptom, or the symptom that afflicts the subject, or at least one underlying cause of a symptom associated with the disease, condition, or symptom that afflicts the subject. Therefore, treatment includes suppressing (e.g., preventing the development or further progression of a disease, condition, or symptom, or its associated clinical symptoms) an active disease.

[0064] As used in this article, "needs treatment" refers to a judgment made by a physician or other caregiver that an individual needs or will benefit from treatment. This judgment is based on a variety of factors within the physician's or caregiver's professional field.

[0065] The terms “prevent,” “preventing,” and “prevention” refer to a process of action initiated in a certain way (e.g., prior to the onset of a disease, condition, symptom, or its symptoms) such as the administration of a CCR6 and / or CXCR2 antagonist or a pharmaceutical composition containing thereto, thereby temporarily or permanently preventing, inhibiting, suppressing, or reducing the risk of a patient developing a disease, condition, or symptom (as determined by the absence of clinical symptoms), or delaying its onset, typically in cases where the subject is predisposed to a particular disease, condition, or symptom. In some cases, these terms also refer to slowing the progression of a disease, condition, or symptom or inhibiting its development into a harmful or other undesirable state.

[0066] As used in this article, "need for prevention" refers to a judgment made by a physician or other caregiver that a subject needs or will benefit from preventive care. This judgment is based on a variety of factors within the physician's or caregiver's professional field.

[0067] The phrase "therapeutic effective dose" refers to the amount of a drug, administered to a subject alone or as part of a pharmaceutical composition, and administered to the subject as a single dose or as part of a series of doses, that has any detectable, positive effect on any symptom, aspect, or condition of the disease, illness, or ailment. Therapeutic effective doses can be determined by measuring the relevant physiological effects and can be adjusted in conjunction with diagnostic analysis of the subject's condition and the dosing regimen. For example, measuring serum levels of CCR6 and / or CXCR2 antagonists (or, for example, their metabolites) at a specific time after administration can indicate whether a therapeutic effective dose has been used.

[0068] The phrase "a quantity sufficient to achieve a change" means a detectable difference between the level of an indicator measured before (e.g., baseline level) and after the administration of a particular therapy. Indicators include any objective parameter (e.g., serum concentration) or subjective parameter (e.g., subject's well-being).

[0069] The term "small molecule" refers to chemical compounds with a molecular weight of less than about 10 kDa, less than about 2 kDa, or less than about 1 kDa. Small molecules include, but are not limited to: inorganic molecules, organic molecules, organic molecules containing inorganic components, molecules containing radioactive atoms, and synthetic molecules. In therapeutic applications, small molecules may be more permeable to cells, less prone to degradation, and less likely to trigger an immune response than large molecules.

[0070] The terms “inhibitor” and “antagonist” or “activator” and “agonist” refer, respectively, to molecules that inhibit or activate, for example, ligands, receptors, cofactors, genes, cells, tissues, or organs. An inhibitor is a molecule that reduces, blocks, prevents, delays, inactivates, desensitizes, or downregulates, for example, genes, proteins, ligands, receptors, or cells. An activator is a molecule that increases, activates, promotes, enhances, sensitizes, or upregulates, for example, genes, proteins, ligands, receptors, or cells. An inhibitor can also be defined as a molecule that reduces, blocks, or inactivates constitutive activity. An “agonist” is a molecule that interacts with a target to cause or promote an increase in target activation. An “antagonist” is a molecule that opposes the action of an agonist. Antagonists prevent, reduce, inhibit, or neutralize the activity of agonists, and antagonists can also prevent, inhibit, or reduce the constitutive activity of a target, such as a target receptor, even in the absence of an identified agonist.

[0071] The terms "modulate" and "modulation" refer to the ability of a molecule (e.g., an activator or inhibitor) to directly or indirectly increase or decrease the function or activity of CCR6 and / or CXCR2. Modulators can act alone, or they can use cofactors such as proteins, metal ions, or small molecules.

[0072] The "activity" of a molecule can describe or refer to the binding of the molecule to the receptor; catalytic activity; the ability to stimulate gene expression or cell signal transduction, differentiation or maturation; antigenic activity; and the regulation of the activity of other molecules, etc.

[0073] As used herein, terms such as “comparable,” “comparable activity,” “comparable activity with,” “comparable effect,” and “comparable effect with” are relative terms that can be observed quantitatively and / or qualitatively. The meaning of these terms generally depends on their context. For example, from a qualitative perspective, two drugs that activate receptors may be considered to have comparable effects, but from a quantitative perspective, such as in assays accepted in the field (e.g., dose-response assays) or in animal models accepted in the field, if one drug achieves only 20% of the activity of the other drug, the two drugs may be considered to lack comparable effects. When comparing one outcome with another (e.g., one outcome with a reference standard), “comparable” often (though not always) means that a result deviates from a reference standard by less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 7%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%. In a specific implementation, a result is considered comparable to the reference standard if its deviation from the reference standard is less than 15%, less than 10%, or less than 5%. By way of example and not limitation, activity or effect can refer to potency, stability, solubility, or immunogenicity.

[0074] "Substantially pure" means that a component constitutes more than about 50% of the total composition, and typically more than about 60%. More typically, "substantially pure" means a composition in which at least 75%, at least 85%, at least 90% or more of the total composition is the target component. In some cases, the target component will constitute more than about 90% or about 95% of the total composition.

[0075] compound

[0076] This article provides compounds having formula (I):

[0077]

[0078] Or any salt, solvate, hydrate, N-oxide, tautomer or rotational isomer thereof,

[0079] in,

[0080] B is selected from the group consisting of furanyl, oxazolyl, phenyl, pyridyl, pyrimidinyl, and pyrazinyl, each optionally modified by R. 1a R 1b and R 2 Replace, R 1a R 1b and R 2 Independently selected from the following groups: halogens, CN, C 1-4 Alkyl, C 1-4Alkoxy and C 1-4 Halogenated alkyl groups;

[0081] R 3 They are members selected from H and D;

[0082] R 4 It is selected from H and C 1-8 Alkyl and Y members; wherein, C 1-8 Alkyl groups are optionally halogenated, -CN, or -CO2R. a -CONR a R b -C(O)R a OC(O)NR a R b -NR a C(O)R b -NR a C(O)2R c -NR a C(O)NR a R b -NR a R b -OR a -S(O)2NR a R b -NR a S(O)2R b Or Y replaces, where each R a and R b Independently selected from: hydrogen, C 1-4 Alkyl, C 1-4 Hydroxyalkyl and C 1-4 Haloalkyl, R c Selected from: C 1-4 Alkyl, C 1-4 Hydroxyalkyl and C 1-4 Haloalkyl; and Y is a 5- or 6-membered aryl or heteroaryl group, optionally selected from 1 to 4 halogens, -CN, -C 1-4 Alkyl, -C 1-4 Alkoxy, -C 1-4 Hydroxyalkyl, -C 1-4 Halogenated alkyl groups, OCF3, -CO2R a -CONR a R b -C(O)R a -OC(O)NR a R b -NR a C(O)R b -CH2CO2R a Substituents of the substituents;

[0083] R 5a and R 5b Each element is independently selected from H, halogen, and C. 1-4 Alkyl, C 1-4 Members of alkoxy, CO2H, and CN;

[0084] R 6a and R 6b They are each independently selected from H and C 1-4 Alkyl, C 1-4 Hydroxyalkyl and C 1-4 Members of haloalkyl groups; or optionally R 6a and R 6b Together they form an oxygen group (=O); and

[0085] The subscript n is 1 or 2.

[0086] In some embodiments, the compound of formula I is a compound wherein B is a furanyl or oxazolyl group, which is optionally converted by R. 1a and R 1b Instead, the R 1a and R 1b Independently selected from the following groups: halogens, CN, C 1-4 Alkyl, C 1-4 Alkoxy and C 1-4 Halogenated alkyl groups. In this set of embodiments, certain selected embodiments are those in which R... 1a Those are CH3. In other alternative implementations, R3 is H. In other alternative implementations, R 5a and R 5b Each of these is independently selected from H, Cl, and F. In other alternative implementations, R 6a and R 6b Each of them is independently selected from H and C. 1-4 Alkyl. In other alternative embodiments, R 4 It is Y. In other alternative implementation schemes, R 4 Selected from H and optionally substituted aryl or heteroaryl groups. In a specific embodiment, R 1a Selected from CH3 and Cl; and R 1b It does not exist or is CH3.

[0087] In a selected set of embodiments, compounds having formula (Ia1) are provided:

[0088]

[0089] Or its pharmaceutically acceptable salt, solvate or hydrate, wherein R 1a Selected from CH3 and Cl; R 1b It does not exist (replaced by H) or CH3; R3 It is H or D; R 4 It is H or optionally substituted aryl or heteroaryl; R 5a and R 5b Each is independently selected from H, F, Cl, Br, and CH3; and R 6a and R 6b Each is independently selected from H and CH3.

[0090] In some embodiments, a compound of formula (Ia1), or a pharmaceutically acceptable salt, solvate, or hydrate thereof, is provided, wherein R 1a It is CH3; R 1b R3 is H or D; R4 is H or an optionally substituted aryl or heteroaryl group; R 5a It is H, Cl, or Br; R 5b It is H or F; and R 6a and R 6b Each is represented by H.

[0091] In some selected embodiments, compounds of formula (I) selected from those compounds in Figure 1 are provided.

[0092] This document also provides compounds having formula (A), or any pharmaceutically acceptable salt, solvate, hydrate, N-oxide, tautomer, or rotational isomer thereof:

[0093]

[0094] in,

[0095] B is selected from the group consisting of furanyl, thiophene, oxazolyl, phenyl, pyridyl, pyrimidinyl, and pyrazinyl, each optionally modified by R. 1a R 1b and R 2 Replace, R 1a R 1b and R 2 Independently selected from the following groups: halogens, CN, C 1-4 Alkyl, C 1-4 Alkoxy and C 1-4 Halogenated alkyl groups;

[0096] R 3 They are members selected from H and D;

[0097] R 4 It is selected from H and C 1-8 Alkyl, OH, -NR a R b -C 1-4 Members of alkoxy and Y; among which, C 1-8Alkyl groups are optionally halogenated, -CN, or -CO2R. a -CONR a R b -C(O)R a OC(O)NR a R b -NR a C(O)R b -NR a C(O)2R c -NR a C(O)NR a R b -NR a R b -OR a -S(O)2NR a R b -NR a S(O)2R b The substituted Y is a 4- to 8-membered cyclohexaalkyl, a 3- to 8-membered cycloalkyl, a 5- or 6-membered aryl, or a heteroaryl, any one of which is optionally replaced by 1 to 4 ions selected from halogen, oxy, -CN, -C 1-6 Alkyl, -C 1-6 Alkoxy, -C 1-6 Hydroxyalkyl, -C 1-6 Halogenated alkyl, OC 1-6 Halogenated alkyl, -C 1-4 Alkyl-OC 1-4 Alkyl, -C 1-6 Alkyl-NR a R b -C 1-6 Alkyl -CO2H, -C 1-6 Alkyl-CO2R a -C 1-6 Alkyl-CONR a R b -C 1-6 Alkyl-C(O)R a -C 1-6 Alkyl-OC(O)NR a R b -C 1-6 Alkyl-NR a C(O)R b -C 1-6 Alkyl-NR a C(O)2R c -C 1-6 Alkyl-NR a C(O)NR a R b -C 1-6 Alkyl-ORa -C 1-6 Alkyl-S(O)2NR a R b -C 1-6 Alkyl-NR a S(O)2R b -CO2R a -CONR a R b -C(O)R a -OC(O)NR a R b -NR a C(O)R b -NR a C(O)2R c -NR a C(O)NR a R b -NR a R b -OR a -S(O)2NR a R b -NR a S(O)2R b -CH2CO2R a Substituents of each R group; a and R b Independently selected from hydrogen and C 1-4 Alkyl, C 1-4 Hydroxyalkyl and C 1-4 Halogenated alkyl groups, and R c Selected from C 1-4 Alkyl, C 1-4 Hydroxyalkyl and C 1-4 Halogenated alkyl; and wherein the 4- to 8-membered heteroalkyl and 3- to 8-membered cycloalkyl groups may optionally be substituted with oxygen groups;

[0098] R 5a and R 5b Each element is independently selected from H, halogen, and C. 1-4 Alkyl, -C 1-4 Halogenated alkyl, OC 1-4 Haloalkyl, C 1-4 Members of alkoxy, CO2H, and CN;

[0099] R 6a and R 6b They are each independently selected from H and C 1-4 Alkyl, C 1-4 Hydroxyalkyl and C 1-4 Members of haloalkyl groups; or optionally R 6a and R6b Together they form an oxygen group (=O) or a 4-6 membered cyclohexaalkyl or a 3-6 membered cycloalkyl;

[0100] R 7 It is selected from methyl, ethyl and C 1-2 Members of haloalkyl groups; and

[0101] The subscript n is 1 or 2.

[0102] In some implementation schemes, B is selected from the following group:

[0103]

[0104] In some implementation schemes, B is selected from the following group:

[0105]

[0106] In some embodiments, B is a furanyl or oxazolyl group, each optionally and independently selected from halogen, CN, C. 1-4 Alkyl, C 1-4 Alkoxy and C 1-4 R of haloalkyl 1a and R 1b replace.

[0107] In some implementations, B is R that is CH3 or Cl. 1a R that replaces and is optionally CH3 1b Substituted furanyl group.

[0108] In some implementation schemes, R 3 It is H.

[0109] In some implementation schemes, R 5a and R 5b Each of them is independently selected from H, CH3, Cl and F.

[0110] In some implementation schemes, Selected from the following group:

[0111]

[0112] In some implementation schemes, Selected from the following group:

[0113]

[0114] In some implementations, each R 6a and R 6b Independently selected from H and C 1-2 alkyl.

[0115] In some implementation schemes, Select independently from the following group

[0116]

[0117] In some implementation schemes, Select independently from the following group

[0118]

[0119] In some implementation schemes, R 4 For H, C 1-3 Alkyl or Y, wherein C 1-3 The alkyl group is replaced by a tetrazolium or tetrazolone group, wherein the tetrazolium or tetrazolone group is optionally C-substituted. 1-6 Alkyl, C 1-6 Hydroxyalkyl or C 1-4 Alkyl-OC 1-4 Alkyl substitution, wherein Y is selected from the group consisting of pyridyl, pyrazolyl, and phenyl, wherein pyridyl, pyrazolyl, and phenyl have 1-3 independently selected from -C. 1-4 Alkyl, -C 1-4 Substituents of alkoxy groups and -CO2H groups.

[0120] In some implementation schemes, R 4 It is H.

[0121] In some implementation schemes, R 4 C 1-6 Alkyl, wherein C 1-6 The alkyl group is replaced by a tetrazolium or tetrazolone group, wherein the tetrazolium or tetrazolone group is optionally -C 1-6 Alkyl, -C 1-6 Alkoxy, -C 1-6 Hydroxyalkyl, -C 1-6 Halogenated alkyl, OC 1-6 Halogenated alkyl, -C 1-4 Alkyl-OC 1-4 Alkyl, -C 1-6 Alkyl-NR a R b -C 1-6 Alkyl -CO2H, -C 1-6 Alkyl-CO2R a -C 1-6 Alkyl-CONR a R b -C 1-6 Alkyl-C(O)R a -C 1-6 Alkyl-OC(O)NR a R b -C 1-6 Alkyl-NRa C(O)R b -C 1-6 Alkyl-NR a C(O)2R c -C 1-6 Alkyl-NR a C(O)NR a R b -C 1-6 Alkyl-OR a -C 1-6 Alkyl-S(O)2NR a R b or -C 1-6 Alkyl-NR a S(O)2R b .

[0122] In some implementation schemes, R 4 C 1-3 Alkyl, wherein C 1-3 The alkyl group is replaced by a tetrazolium or tetrazolone group, wherein the tetrazolium or tetrazolone group is optionally -C 1-6 Alkyl, -C 1-6 Alkoxy, -C 1-6 Hydroxyalkyl, -C 1-6 Halogenated alkyl, -C 1-4 Alkyl-OC 1-4 Alkyl, -C 1-6 Alkyl-NR a R b or -C 1-6 Alkyl-CO2H.

[0123] In some implementation schemes, R 4 C 1-3 Alkyl, wherein C 1-3 The alkyl group is replaced by a tetrazolium or tetrazolone group, wherein the tetrazolium or tetrazolone group is optionally C-substituted. 1-3 Alkyl, C 1-3 Hydroxyalkyl or C 1-3 Alkyl-OC 1-3 Alkyl substitution.

[0124] In some implementation schemes, R 4 Selected from pyridyl, pyrazolyl, and phenyl, wherein pyridyl, pyrazolyl, and phenyl have 1-3 independently selected from -C 1-4 Alkyl, -C 1-4 Substituents of alkoxy groups and -CO2H groups.

[0125] In some implementation schemes, R 4 C is substituted with a tetrazolium or tetrazolone group. 1-3 Alkyl group, wherein the tetrazolium or tetrazolone group is optionally C1-3 Alkyl substitution.

[0126] In some implementation schemes, R 4 Selected from the following group:

[0127]

[0128] In some implementation schemes, R 7 Selected from methyl, ethyl, and CF3. In some embodiments, R 7 It is a methyl group.

[0129] In some embodiments, a compound of formula (A1), or a pharmaceutically acceptable salt, solvate, or hydrate thereof, is provided:

[0130]

[0131] Where R 1a Selected from CH3 and Cl; R 1b It does not exist or is CH3; R 3 It is H or D; R 4 It is H or Y; R 5a and R 5b Each is independently selected from H, F, Cl, Br, and CH3; R 6a and R 6b Each is independently selected from H and CH3; and R 7 It is methyl or ethyl.

[0132] In some implementation schemes, R 1a It is CH3; R 1b It does not exist or is CH3; R 3 It is H or D; R 4 It is H; R 5a It is H, F, Me, or Cl or Br; R 5b It is H or F; R 6a and R 6b Each is H; and R 7 It is methyl or ethyl; or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0133] In some embodiments, the compound carries R 3 The carbon atoms are essentially free of other isomers.

[0134] In some implementation schemes, R 4 It's Y.

[0135] In some embodiments, a compound of formula (A2), or a pharmaceutically acceptable salt, solvate, or hydrate thereof, is provided:

[0136]

[0137] Where R 1a Selected from CH3 and Cl; R 1b It is H or CH3; R 3 It is H or D; R 4a and R 4b Independently selected from halogens, -CN, -C 1-4 Alkyl, -C 1-4 Alkoxy, -C 1-4 Hydroxyalkyl, -C 1-4 Halogenated alkyl groups, OCF3,-CO2R a -CONR a R b -C(O)R a -OC(O)NR a R b -NR a C(O)R b -CH2CO2R a , and R a and R b Independently selected from hydrogen and C 1-4 Alkyl, C 1-4 Hydroxyalkyl and C 1-4 Halogenated alkyl; R 5a and R 5b Each is independently selected from H, F, Cl, Br, and CH3; R 6a and R 6b Each is independently selected from H and CH3; and R 7 Selected from methyl, ethyl and C 1-2 Halogenated alkyl groups.

[0138] In some embodiments, compounds selected from the group consisting of or pharmaceutically acceptable salts thereof are provided:

[0139]

[0140] In some embodiments, compounds selected from the group consisting of or pharmaceutically acceptable salts thereof are provided:

[0141]

[0142]

[0143] In some embodiments, compounds selected from the group consisting of or pharmaceutically acceptable salts thereof are provided:

[0144]

[0145] Preparation of compounds

[0146] The embodiments described below provide certain synthetic routes that can be followed to obtain certain compounds of the present invention. Other routes or modifications to the routes described below will be apparent to those skilled in the art and are all within the scope of protection of the present invention.

[0147] Pharmaceutical Composition

[0148] In addition to the compounds provided above, compositions for modulating CCR6 and / or CXCR2 activity in humans and animals typically contain a drug carrier or diluent.

[0149] As used herein, the term "composition" shall include products containing specific amounts of specific ingredients, and any products produced directly or indirectly from combinations of specific amounts of specific ingredients. "Pharmaceutically acceptable" means that the carrier, diluent, or excipient must be compatible with the other components of the formulation and harmless to its recipients.

[0150] Pharmaceutical compositions for administering the compounds of the present invention can be conveniently available in unit dosage forms and can be prepared by any method well known in the medical and pharmaceutical delivery fields. All methods involve the step of binding the active ingredient with a carrier comprising one or more auxiliary components. Pharmaceutical compositions are generally prepared by uniformly and tightly binding the active ingredient with a liquid carrier or a subdivided solid carrier, or both, and then, if desired, forming the product into the desired formulation. In the pharmaceutical composition, the amount of the active target compound is sufficient to produce the desired effect on the course or condition of a disease.

[0151] Pharmaceutical compositions containing an active ingredient can be in forms suitable for oral administration, such as tablets, lozenges, gummies, aqueous or oily suspensions, dispersible powders or granules, emulsions and self-emulsifying solutions (as described in U.S. Patent Application 6,451,339), hard or soft capsules, syrups, elixirs, solutions, oral patches, oral gels, chewable gums, chewable tablets, effervescent powders, and effervescent tablets. Compositions for oral use can be prepared according to any method known in the field of pharmaceutical composition preparation. Such compositions may contain one or more agents selected from sweeteners, flavoring agents, coloring agents, antioxidants, and preservatives to provide a pharmaceutically superior and palatable product. Tablets contain the active ingredient mixed with non-toxic, pharmaceutically acceptable excipients suitable for tablet preparation. These excipients can be, for example, inert diluents such as cellulose, silica, alumina, calcium carbonate, sodium carbonate, glucose, mannitol, sorbitol, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrants such as corn starch or alginic acid; binders such as PVP, cellulose, PEG, starch, gelatin, or gum arabic; and lubricants such as magnesium stearate, stearic acid, or talc. These tablets can be uncoated or they can be enteric- or otherwise coated using known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing sustained action over a longer period. For example, delaying materials such as glyceryl monostearate or glyceryl distearate can be used. They can also be coated using the techniques described in U.S. Patent Nos. 4,256,108; 4,166,452 and 4,265,874 to form osmotic therapeutic tablets for controlled release.

[0152] Oral formulations can also be hard gelatin capsules, in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin, or soft gelatin capsules, in which the active ingredient is mixed with water or an oily medium, such as peanut oil, liquid paraffin, or olive oil. Furthermore, emulsions can be prepared using non-aqueous miscible ingredients, such as oils, and stabilized with surfactants such as monoglycerides, diglycerides, PEG esters, etc.

[0153] Aqueous suspensions contain active ingredients mixed with excipients suitable for preparing aqueous suspensions. Such excipients are suspending agents, such as sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and gum arabic; dispersants or wetting agents may be naturally occurring phospholipids, such as lecithin, or condensation products of alkyl esters and fatty acids, such as polyoxyethylene stearate, or condensation products of ethylene oxide and long-chain fatty alcohols, such as heptadecaethyleneoxycetanol, or condensation products of ethylene oxide and esters derived from fatty acids and hexitols, such as polyoxyethylene sorbitan monooleate, or condensation products of ethylene oxide and esters derived from fatty acids and hexitol anhydrides, such as polyvinyl dehydrated sorbitan monooleate. Aqueous suspensions may also contain one or more preservatives, such as ethylparaben or n-propylparaben, one or more colorants, one or more flavorings, and one or more sweeteners, such as sucrose or saccharin.

[0154] Oily suspensions are formulated by suspending active ingredients in vegetable oils such as peanut oil, olive oil, sesame oil, or coconut oil, or in mineral oils such as liquid paraffin. Oily suspensions may contain thickeners such as beeswax, hard paraffin, or cetyl alcohol. Sweeteners and flavorings, as described above, may also be added to provide palatable oral products. These compositions may be preserved by adding antioxidants such as ascorbic acid.

[0155] Dispersible powders and granules suitable for preparing aqueous suspensions by adding water provide an active ingredient that can be mixed with a dispersant or wetting agent, a suspending agent, and one or more preservatives. Examples of suitable dispersants or lubricants and suspending agents are those mentioned above. Other excipients, such as sweeteners, flavoring agents, and coloring agents, may also be present.

[0156] The pharmaceutical compositions 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, or a mineral oil, such as liquid paraffin or a mixture thereof. Suitable emulsifiers may be naturally occurring gums, such as gum arabic or tragacanth gum, naturally occurring phospholipids, such as soybean, lecithin, and esters or metaesters derived from fatty acids and hexitan anhydrides, such as sorbitan monooleate, and condensates of said metaesters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners and flavoring agents.

[0157] Syrups and elixirs can be formulated with sweeteners such as glycerin, propylene glycol, sorbitol, or sucrose. These formulations may also contain modifiers, preservatives, flavoring agents, and / or coloring agents. Oral solutions can be prepared in combination with, for example, cyclodextrin, PEG, and surfactants.

[0158] Pharmaceutical compositions may be in the form of sterile injectable aqueous or oily suspensions. Such suspensions may be formulated using suitable dispersants or wetting agents and suspending agents as described above, according to known techniques. Sterile injectable products may also be sterile injectable solutions or suspensions formulated with non-toxic, parenteral-acceptable diluents or solvents, such as 1,3-butanediol solutions. Water, Ringer's solution, and isotonic sodium chloride solution may be used among acceptable carriers and solvents. Furthermore, sterile, non-volatile oils are routinely used as solvents or suspension media. For this purpose, any mild, non-volatile oil may be used, including synthetic mono- or diglycerides. Additionally, fatty acids, such as oleic acid, may be used in the preparation of injectable formulations.

[0159] The compounds of the present invention can also be administered rectally in suppository form. 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, thereby melting and releasing the drug within the rectum. Such substances include coconut oil and polyethylene glycol. Furthermore, the compounds can be delivered via ocular delivery using solutions or ointments. Transdermal administration of the compounds of the present invention can also be achieved using iontophoresis patches, etc. For topical use, creams, ointments, gels, solutions, or suspensions containing the compounds of the present invention can be utilized. As used herein, topical application is also intended to include the use of mouthwashes and mouthwashes.

[0160] The compounds of this invention can be formulated within a medical device, which may include any variety of conventional grafts, stents, including covered stents, catheters, balloons, baskets, or other devices that can be configured or permanently implanted into a body cavity. As in a particular embodiment, it is desirable to have a device or method capable of delivering the compounds of this invention to a body site that has already undergone interventional treatment.

[0161] In an exemplary embodiment, the inhibitor of the present invention may be placed in a medical device, such as a stent, and delivered to a treatment site in a part of the body for treatment.

[0162] Stents have been used as carriers for delivering therapeutic agents (i.e., drugs). Intravascular stents are typically permanently implanted in coronary or peripheral blood vessels. Stent designs include those in U.S. Patent Nos. 4,733,655 (Palmaz), 4,800,882 (Gianturco), or 4,886,062 (Wiktor). These designs include metallic and polymer stents, as well as self-expanding and balloon-expanding stents. The stents can also be used to deliver drugs at sites of contact with blood vessels, as disclosed in U.S. Patent Nos. 5,102,417 (Palmaz) and international patent applications WO91 / 12779 (Medtronic, Inc.) and WO90 / 13332 (Cedars-Sanai Medical Center), U.S. Patent Nos. 5,419,760 (Narciso, Jr.) and 5,429,634 (Narciso, Jr.). The scaffold is also used to deliver viruses to the inner wall of the cavity for gene delivery, as disclosed in U.S. Patent 5,833,651 (Donovan et al.).

[0163] In one embodiment, the inhibitor may be incorporated into the polymer composition during the formation of a biocompatible coating on a medical device (e.g., a stent). The coatings obtained from these components are typically uniform and can be used to coat some implantable devices.

[0164] The polymer can be non-degradable or bioabsorbable, depending on the desired release rate or desired polymer stability. However, bioabsorbable polymers are preferred because, unlike non-degradable polymers, they do not persist long after implantation, thus avoiding any adverse reactions or chronic local reactions. Usable bioabsorbable polymers include, but are not limited to: poly(L-lactic acid), polycaprolactone, polyglycolic acid (PGA), poly(lactide-co-glycolic acid) (PLLA / PGA), poly(hydroxybutyrate), poly(hydroxybutyrate-co-valerate), polydioxanone, polyorthoester, polyanhydride, poly(glycolic acid), poly(D-lactic acid), poly(L-lactic acid), poly(D,L-lactic acid), poly(D,L-lactide) (PLA), poly(L-lactide) (PLLA), poly(glycolic acid-co-trimethylene carbonate) (PGA / PTMC), and polyethylene oxide (PEO). Poly(dioxanone) (PDS), polyphosphates, polyphosphate urethane, poly(amino acids), cyanoacrylates, poly(trimethylene carbonate), poly(imino carbonate), copoly(ether-ester) (e.g., PEO / PLA), polyalkylene oxalates, polyphosphazenes and biomolecules (e.g., fibrin, fibrinogen, cellulose, starch, collagen and hyaluronic acid), poly(ε-caprolactone), polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyran, polycyanoacrylates, crosslinked or amphiphilic block copolymers of hydrogels, and other suitable bioabsorbable polymers known in the art. In addition, non-degradable polymers with relatively low chronic tissue reactions, such as polyurethanes, silicones, and polyesters, can be used; other polymers can also be used if they can be dissolved, cured, or polymerized on the medical device, such as polyolefins, polyisobutylene and ethylene-α-olefin copolymers, acrylic polymers and copolymers, vinyl halide polymers and copolymers, such as polyvinyl chloride; polyvinylpyrrolidone; polyvinyl ethers, such as polyvinyl methyl ether; polyvinylidene halides, such as polyvinylidene fluoride and polyvinylidene chloride; polyacrylonitrile, polyvinyl ketone; polyvinyl aromatic hydrocarbons, such as polystyrene, polyethylene esters, such as polyvinyl acetate; and copolymers of vinyl monomers. And, olefins, such as ethylene-methyl methacrylate copolymers, acrylonitrile-styrene copolymers, ABS resins and ethylene-vinyl acetate copolymers; pyran copolymers, polyhydroxypropyl methacrylamide-phenol; polyhydroxyethyl-asparagine-phenol; palmitoyl residue-substituted polyethylene oxide-polylysine; polyamides, such as nylon 66 and polycaprolactam; alkyd resins, polycarbonates; polyoxymethylene; polyimide; polyether; epoxy resins, polyurethane; synthetic fibers; synthetic fiber-triacetate; cellulose, cellulose acetate, cellulose butyrate; cellulose acetate butyrate; cellophane; nitrocellulose; cellulose propionate; cellulose ethers; and carboxymethyl cellulose.

[0165] Polymer and semi-permeable polymer matrices can be processed into molded products, such as valves, stents, catheters, and prostheses.

[0166] In one embodiment of the invention, the inhibitor of the invention is coupled to a polymer or a semi-permeable polymer matrix, the semi-permeable polymer matrix being formed into a scaffold or a scaffold-type graft device.

[0167] Typically, polymers can be applied to the surface of implantable devices by spin coating, dipping, or spraying. Other methods known in the art can also be used for this purpose. Spraying methods include conventional methods as well as microdeposition techniques with inkjet dispensers. Additionally, the polymer can be deposited onto the implantable device using image graphics, such that the polymer is placed only on specific portions of the device. The coating on the device provides a uniform layer surrounding the device, taking into account the enhanced diffusion of different analytes through the device coating.

[0168] In a preferred embodiment of the invention, the inhibitor is formulated for release from the polymer coating into the environment in which the medical device is placed. Preferably, the inhibitor is released in a controlled manner over an extended time period (e.g., months) by using at least one of several known techniques involving polymer carriers or coatings for controlled elution. Some of these techniques have previously been described in U.S. Patent Application 20040243225A1.

[0169] Furthermore, as described in U.S. Patent 6,770,729, the reagents and reaction conditions of the polymer composition are controllable to control the release of inhibitors from the polymer coating. For example, the diffusion coefficients of one or more polymer coatings can be adjusted to control the release of inhibitors from the polymer coating. In variations of this subject matter, the diffusion coefficients of one or more polymer coatings can be controlled to adjust the levels of analytes present in the environment in which the medical device is placed (e.g., analytes that promote the decomposition or hydrolysis of a portion of the polymer) to obtain one or more components of the polymer composition (e.g., thereby modulating the release of inhibitors from the polymer coating). However, another embodiment of the invention includes a device having multiple polymer coatings, each coating having multiple diffusion coefficients. In such embodiments of the invention, the release of inhibitors from the polymer coatings can be achieved through multiple polymer coatings.

[0170] In another embodiment of the invention, the release of the inhibitor from the polymer coating is controlled by adjusting one or more properties of the polymer composition, such as the presence of one or more endogenous or exogenous compounds, or the pH of the polymer composition. For example, certain polymer compositions may be designed to release the inhibitor in response to a decrease in the pH of the polymer composition. Alternatively, certain polymer compositions may be designed to release the inhibitor in response to the presence of hydroperoxide.

[0171] In some embodiments, pharmaceutical compositions comprising the compounds of the present invention are provided. In some embodiments, the pharmaceutical composition further comprises one or more additional therapeutic agents. In some embodiments, the one or more additional therapeutic agents are selected from cytotoxic chemotherapy, anticancer or antitumor vaccines, anti-immunocytokine therapy, immunocytokine therapy, chimeric antigen receptor (CAR) T-cell receptors, gene transfer therapy, checkpoint inhibitors, corticosteroids, retinoid-like agents, antitumor drugs, and interferon analogs. In some embodiments, the one or more additional therapeutic agents are selected from the group consisting of: TNFα ligand inhibitors, TNF binders, IL-1 ligand inhibitors, IL-6 ligand inhibitors, IL-8 ligand inhibitors; IL-17 antagonists, calcineurin inhibitors, TNF antagonists, retinoic acid receptor γ antagonists, IL-17A ligand inhibitors; IL-17F ligand inhibitors, RIP-1 kinase inhibitors, sphingosine-1-phosphate receptor-1 antagonists, sphingosine-1-phosphate receptor-1 modulators, Rho-associated protein kinase 2 inhibitors, IL-12 antagonists; IL-23 antagonists, type II TNF receptor modulators, IL-23A inhibitors, PDE4 inhibitors, JAK tyrosine kinase inhibitors, Jak1 tyrosine kinase inhibitors; Jak3 tyrosine kinase inhibitors, histamine H1 receptor antagonists, retinoic acid receptor agonists, copper amine oxidase inhibitors, PI3K modulators, and phosphatidylinositol-3 kinase δ inhibitors. Agents, mitochondrial 10kDa heat shock protein stimulators, adenosine A3 receptor agonists, galactolectin-3 inhibitors, F1F0 ATP synthase modulators, GM-CSF ligand inhibitors, vitamin D3 receptor agonists, glucocorticoid agonists, histamine H4 receptor antagonists, CCR3 chemokine antagonists, eosinophil activation chemokine ligand inhibitors, sphingosine-1-phosphate receptor-1 modulators, phospholipase A2 inhibitors, PDE4 inhibitors, albumin modulators, TLR-7 antagonists, TLR-8 antagonists, TLR-9 antagonists, CD40 ligand receptor antagonists, Src tyrosine kinase inhibitors, tubulin binders, interleukin-1α ligand inhibitors, histone deacetylase-1 inhibitors, histone deacetylase-2 inhibitors, histone deacetylase-3 inhibitors, histone deacetylase-6 inhibitors, nucleoside reverse transcriptase inhibitors, nuclear factor κB inhibitors, STAT-3 inhibitors, parathyroid hormone ligand inhibitors;Vitamin D3 receptor agonists, T cell surface glycoprotein CD28 stimulators, histamine H4 receptor antagonists, TGFβ agonists, P-selectin glycoprotein ligand-1 stimulators, DHFR inhibitors, retinoic acid receptor γ modulators, cytoplasmic phospholipase A2 inhibitors, visual pigment X receptor modulators, β-catenin inhibitors, CREB-binding protein inhibitors, TrkA receptor antagonists, T cell differentiation antigen CD6 inhibitors, ADP-ribosylcyclase-1 inhibitors, interleukin-1β ligand modulators; insulin receptor substrate-1 inhibitors, DHFR inhibitors, IL-8 antagonists, CTLA-4 (CD15) blockers 2) Drugs containing PD-1 (CD279), PDL-1 (CD274), TIM-3, LAG-3 (CD223), VISTA, KIR, NKG2A, BTLA, PD-1H, TIGIT, CD96, 4-1BB (CD137), 4-1BBL (CD137L), GARP, CSF-1R, A2AR, CD73, CD47, tryptophan 2,3-dioxygenase (TDO) or indoleamine 2,3-dioxygenase (IDO) activity, and agonists of OX40, GITR, 4-1BB, ICOS, STING, or CD40.

[0172] Treatment of diseases regulated by CCR6 and / or CXCR2

[0173] On one hand, the present invention provides a method for treating or preventing CCR6-mediated symptoms or diseases and / or CXCR2-mediated symptoms or diseases, said method administering a therapeutically effective amount of any compound of the present invention to a subject having such symptoms or diseases. Preferred compounds for use in this method are those compounds described herein as preferred embodiments, as well as compounds specifically listed in the examples below; and compounds with specific structures provided herein. Hereinafter, "subject" is defined to include animals, such as mammals, including but not limited to primates (e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, and the like. In a preferred embodiment, the subject is a human.

[0174] As used herein, the phrase "CCR6-mediated condition or disease" and related phrases and terms refer to a condition or disease characterized by inappropriate (e.g., less than or greater than normal) CCR6 functional activity. Inappropriate CCR6 functional activity can result from the expression of CCR6 by cells that normally do not express CCR6, increased CCR6 expression (leading to, for example, inflammatory and immune regulatory disorders and diseases), or decreased CCR6 expression. Inappropriate CCR6 functional activity can also result from the secretion of CCL20 by cells that normally do not secrete CCL20, increased CCL20 expression (leading to, for example, inflammatory and immune regulatory disorders and diseases), or decreased CCL20 expression. Inappropriate CCR6 functional activity can mediate CCR6-mediated conditions or diseases entirely or partially. However, CCR6-mediated conditions or diseases are those that modulate CCR6 to produce some therapeutic effect on an underlying condition or disease (e.g., CCR6 antagonists result in some improvement in patient comfort in at least some patients).

[0175] Similarly, the phrase "CXCR2-mediated symptom or disease" and related phrases and terms refer to a symptom or disease characterized by inappropriate, such as less or more than normal, CXCR2 functional activity. Inappropriate CXCR2 functional activity may result from CXCR2 expression in cells that normally do not express CXCR2, increased CXCR2 expression (leading to, for example, inflammation and immune dysregulation and disease), or decreased CXCR2 expression. CXCR2-mediated symptom or disease may be mediated entirely or partially by inappropriate CXCR2 functional activity. However, CXCR2-mediated symptom or disease is one in which CXCR2 modulates to produce some therapeutic effect on an underlying symptom or disease (e.g., CXCR2 antagonists result in some improvement in patient comfort in at least some patients).

[0176] The term "therapeutic effective dose" refers to the amount of a target compound that will elicit a biological or medical response in a tissue, system, animal, or human being sought by researchers, veterinarians, physicians, or other clinicians.

[0177] Diseases and conditions associated with inflammation, infection, and cancer can be treated or prevented using the compounds and compositions of the present invention. In one set of embodiments, diseases or conditions in humans or other species, including chronic diseases, can be treated with inhibitors of CCR6 function. These diseases or conditions include: (1) allergic diseases, such as systemic anaphylactic or hypersensitivity reactions, drug allergies, insect bite allergies, and food allergies; (2) inflammatory bowel diseases, such as Crohn's disease, ulcerative colitis, ileitis, and enteritis; (3) vaginitis; (4) psoriasis and inflammatory skin diseases, such as dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, urticaria and pruritus, vitiligo; (5) vasculitis; (6) spondyloarthritis; (7) scleroderma; (8) asthma and respiratory allergic diseases, such as allergic asthma, allergic rhinitis, hypersensitivity lung disease, etc.; and (9) autoimmune diseases. Diseases such as arthritis (including rheumatoid and psoriatic arthritis), and such as Hashimoto's thyroiditis and Graves' disease, multiple sclerosis, systemic lupus erythematosus, type I diabetes, glomerulonephritis, etc., (10) graft rejection (including allogeneic graft rejection and graft-versus-host disease), (11) other diseases requiring suppression of adverse inflammation, such as atherosclerosis, myositis, neurodegenerative diseases (e.g., Alzheimer's disease), encephalitis, meningitis, hepatitis, nephritis, sepsis, sarcoidosis, allergic conjunctivitis, otitis, chronic obstructive pulmonary disease, sinusitis, Behcet's syndrome and gout.

[0178] Preferably, the method relates to the treatment of diseases or conditions selected from the group consisting of: allergic diseases, psoriasis, skin conditions (such as atopic dermatitis), and asthma and scleroderma.

[0179] In another set of embodiments, CCR6-dependent regulatory T-cell transport can be modulated to treat diseases or conditions including: cancer, infectious diseases (viral infections, such as HIV infection, and bacterial infections), and immunosuppressive diseases, such as organ transplantation status and skin transplantation status. The term "organ transplantation status" is intended to include bone marrow transplantation status and solid organ (e.g., kidney, liver, lung, heart, pancreas, or combinations thereof) transplantation status.

[0180] Given that they inhibit the binding of CXCR2, the compounds of the present invention can be used to treat CXCR2-mediated conditions or diseases, such as inflammatory or allergic conditions or diseases, particularly chronic obstructive pulmonary airway or lung diseases (COPD, COAD, or COLD), including chronic bronchitis or related dyspnea, emphysema, bronchiolitis obliterans syndrome, and severe asthma.

[0181] The compounds of this invention can also be used to treat various diseases, such as cancers, including colorectal cancer, ovarian cancer, prostate cancer, melanoma including metastatic melanoma, lung cancer (e.g., non-small cell lung cancer), renal cell carcinoma; tumor angiogenesis, ischemia / reperfusion injury, delayed graft function, osteoarthritis, myelofibrosis, adenomyosis, contact allergies (skin), and wound healing. Treatments according to the invention can be symptomatic or preventative.

[0182] The preventative effects of treating chronic bronchitis or COPD will be demonstrated by reducing the frequency or severity of symptoms, providing symptom relief and slowing disease progression, and improving lung function. This can be further demonstrated by reducing the need for other symptomatic treatments (i.e., treatments intended to limit or discontinue symptom episodes, such as anti-inflammatory drugs (e.g., corticosteroids) or bronchodilators).

[0183] Other inflammatory or obstructive airway diseases and conditions to which this invention is applicable include acute lung injury (ALI), acute / adult respiratory distress syndrome (ARDS), idiopathic pulmonary fibrosis, fibrotic lung, airway hyperresponsiveness, dyspnea, pulmonary fibrosis, allergic airway inflammation, small airway disease, lung cancer, acute chest syndrome in patients with sickle cell disease and pulmonary hypertension, and exacerbations of airway hyperresponsiveness due to other drug treatments (especially other inhaled drug treatments). This invention is also applicable to the treatment of bronchitis of any type or cause, including, for example, acute, peanut inhalational, catarrhal, croupus, chronic, or tuberculous bronchitis. Further inflammatory or obstructive airway diseases to which this invention applies include pneumoconiosis of any type or cause (inflammatory, often occupational lung disease, whether chronic or acute, often accompanied by airway obstruction and caused by repeated inhalation of dust), including, for example, alumina pneumoconiosis, carbon monoxide pneumoconiosis, asbestosis, stone dust pneumoconiosis, ostrich feather pneumoconiosis, pulmonary iron dust pneumoconiosis, silicosis, smoke pneumoconiosis, and cotton dust pneumoconiosis.

[0184] The compounds of this invention can also be used to treat respiratory viral infections that exacerbate underlying chronic conditions such as asthma, chronic bronchitis, COPD, otitis media, and sinusitis. The treated respiratory viral infections may be associated with secondary bacterial infections, such as otitis media, sinusitis, or pneumonia.

[0185] The compounds of this invention can also be used to treat inflammatory skin conditions such as psoriasis, atopic dermatitis, lupus erythematosus, and other inflammatory or allergic skin conditions.

[0186] The compounds of this invention can also be used to treat other diseases or conditions, particularly those with inflammatory components, such as diseases affecting the nose (including allergic rhinitis, such as atrophic, chronic, or seasonal rhinitis), inflammatory conditions of the gastrointestinal tract (such as inflammatory bowel diseases, such as ulcerative colitis and Crohn's disease), bone and joint diseases (including rheumatoid arthritis, psoriatic arthritis), and other diseases such as atherosclerosis, multiple sclerosis, and acute and chronic allogeneic transplant rejection (e.g., after heart, kidney, liver, lung, or bone marrow transplantation).

[0187] The compounds of this invention can also be used to treat endotoxic shock, glomerulonephritis, cerebral and cardiac ischemia, Alzheimer's disease, cystic fibrosis, viral infections and their associated exacerbations, acquired immunodeficiency syndrome (AIDS), multiple sclerosis (MS), Helicobacter pylori-associated gastritis, and cancer (especially the growth of ovarian cancer).

[0188] The compounds of this invention can also be used to treat symptoms caused by viral infections in humans, including those caused by human rhinovirus, other enteroviruses, coronaviruses, herpesviruses, influenza viruses, parainfluenza viruses, respiratory syncytial viruses, or adenoviruses. The compounds of this invention can also be used to treat pancreatitis.

[0189] The effectiveness of the compounds of the present invention in inhibiting inflammatory conditions such as inflammatory airway diseases can be demonstrated in animal models of airway inflammation or other inflammatory conditions (e.g., mouse, rat, or rabbit models), as described, for example, by Wada et al., J. Exp. Med 180: 1135-40 (1994); Sekido et al., Nature 365: 654-57 (1993); Modelska et al., Am. J. Respir. Crit. Care. Med 160: 1450-56 (1999); and Laffon et al., Am. J. Respir. Crit. Care. Med. 160: 1443-49 (1999).

[0190] In some embodiments, this document provides methods for treating CXCR2-mediated conditions or diseases, such as inflammatory or allergic conditions, particularly inflammatory or obstructive airway diseases, comprising administering to a desired subject (particularly a human) an effective amount of a compound of formula (A), (A1), (A2), (I), or (Ia1) in its free form or pharmaceutically acceptable salt form as described above. In another aspect, the invention provides the use of compounds of formula (A), (A1), (A2), (I), or (Ia1) as described above, in their free form or pharmaceutically acceptable salt form, for the preparation of medicaments for treating CXCR2-mediated conditions or diseases (e.g., inflammatory or allergic conditions or diseases, particularly inflammatory or obstructive airway diseases).

[0191] The compounds of formulas (A), (A1), (A2), (I), and (Ia1) described herein can also be used as co-therapeutic compounds in combination with other drugs (e.g., anti-inflammatory drugs, bronchodilators, antihistamines, or antitussives), particularly for treating obstructive or inflammatory airway diseases such as those mentioned above, for example, as enhancers of the therapeutic activity of such drugs or as a means of reducing the required dosage or potential side effects of these drugs. The compounds of the present invention can be mixed with other drugs in a fixed pharmaceutical composition, or they can be administered alone before, simultaneously with, or after another drug.

[0192] Depending on the disease to be treated and the condition of the patient, the compounds of the present invention can be administered orally, parenterally (e.g., intramuscular, intraperitoneal, intravenous, ICV, intracisional injection or infusion, subcutaneous injection or implantation), by inhalation, nasal, transthelial, rectal, sublingual, or local administration routes. They can be formulated alone or together with suitable dosage units containing conventional, non-toxic, pharmaceutically acceptable carriers, adjuvants, and delivery vehicles suitable for each route of administration. The present invention also contemplates the use of long-acting formulations to administer the compounds of the present invention.

[0193] Those skilled in the art will understand that agents modulating CCR6 activity can be combined with other therapeutic agents and / or chemotherapeutic agents or radiation in a treatment regimen. In some cases, the dosage of chemotherapeutic agents or radiation will be subtherapeutic if not provided with the compositions of the present invention. Those skilled in the art will recognize that "combination" can refer to a combination of treatment methods (i.e., two or more drugs administered as a mixture, or introduced into the subject at least simultaneously or at least at different times, but such that both are simultaneously in the subject's bloodstream). Additionally, the compositions of the present invention can be administered before or after a second treatment regimen, for example, before or after a dose of chemotherapy or radiation.

[0194] The compounds of this invention can be used accordingly for the prevention and treatment of various inflammatory and immune dysregulation disorders and diseases.

[0195] In the treatment or prevention of conditions requiring chemokine receptor regulation, appropriate dosage levels are typically about 0.001 to 100 mg / kg of patient body weight per day, which may be administered in one or more doses. Preferably, the dosage level is about 0.01 to about 25 mg / kg per day; more preferably, about 0.05 to about 10 mg / kg per day. Suitable dosage levels may be about 0.01 to 25 mg / kg per day, about 0.05 to 10 mg / kg per day, or about 0.1 to 5 mg / kg per day. Within this range, the dosage may be 0.005 to 0.05, 0.05 to 0.5, or 0.5 to 5.0 mg / kg per day. For oral administration, the composition is preferably given in tablet form, the tablets containing 1.0 to 1000 mg 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 mg of active ingredient, for dose adjustment to the condition of the patient being treated. The composition can be administered in a regimen of 1 to 4 times daily, preferably once or twice daily.

[0196] However, it should be understood that the specific dose level and frequency for any particular patient can vary depending on a variety of factors, including the activity of the specific compound used, the metabolic stability and duration of action of the compound, the subject's age, weight, genetic characteristics, general health status, sex and diet, as well as the route and timing of administration, excretion rate, drug combination and the severity of the specific condition of the subject being treated.

[0197] Diseases and conditions related to inflammation, immune disorders, infections, and cancer can be treated or prevented using the compounds, compositions, and methods of the present invention.

[0198] The compounds and compositions described in this invention can be used in combination with other compounds and compositions having relevant uses to prevent and treat conditions and diseases of interest, such as inflammation or autoimmune disorders, conditions and diseases including inflammatory bowel disease, rheumatoid arthritis, osteoarthritis, psoriatic arthritis, polyarthritis, multiple sclerosis, allergic diseases, psoriasis, atopic dermatitis and asthma, as well as those conditions mentioned above.

[0199] For example, in the treatment or prevention of inflammatory or autoimmune diseases, or arthritis such as osteoporosis-related arthritis, the compounds and compositions of the present invention may be used with anti-inflammatory agents or analgesics (such as opioid agonists, lipoxygenase inhibitors (such as 5-lipoxygenase inhibitors), cyclooxygenase inhibitors (such as cyclooxygenase-2 inhibitors), interleukin inhibitors (such as interleukin-1 inhibitors), NMDA antagonists, nitric oxide inhibitors or nitric oxide synthesis inhibitors, nonsteroidal anti-inflammatory agents, or cytokine-inhibiting anti-inflammatory agents), such as with compounds (such as acetaminophen, aspirin, codeine, fentanyl, ibuprofen, indomethacin, ketorolac, morphine, naproxen, phenacetin, piroxicam, steroidal analgesics, sufentanil, sullinic acid, tenidap, etc.). Similarly, the compounds and compositions of the present invention can be administered with the analgesics listed above; with synergists such as caffeine, H2 antagonists (e.g., ranitidine), dimethicone, aluminum hydroxide, or magnesium hydroxide; with decongestants such as phenylephrine, phenylpropanolamine, pseudoephedrine, oxymetazoline, epinephrine, naphazoline, xylometazoline, hexahydrodeoxyephedrine, or levonorgestrel; with antitussives such as codeine, dihydrocodeinone, dextromethorphan, dextromethorphan, or dextromethorphan; with diuretics; and with sedative or non-sedative antihistamines.

[0200] Similarly, the compounds and compositions of the present invention can be used in combination with other pharmaceutical products used to treat, prevent, inhibit, or improve diseases or conditions for which the compounds and compositions of the present invention are useful. Such other pharmaceutical products can be administered simultaneously or sequentially with the compounds or compositions of the present invention via the usual route and dosage. When the compounds or compositions of the present invention are used simultaneously with one or more other pharmaceutical products, pharmaceutical compositions containing such other pharmaceutical products in addition to the compounds or compositions of the present invention are preferred. Therefore, pharmaceutical compositions of the present invention include those containing 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 can be used in combination with the compounds or compositions of the present invention, administered separately or in the same pharmaceutical composition, include, but are not limited to: (a) VLA4 antagonists; (b) corticosteroids, such as clotrimazole, prednisolone, betamethasone, prednisone, dexamethasone, fluticasone, hydrocortisone, budesonide, triamcinolone, salmeterol, salmeterol, salmeterol, salbutamol, and formoterol; (c) immunosuppressants, such as cyclosporine (cyclosporine A, cyclosporine B, cyclosporine C, cyclosporine D ... ), Tacrolimus (FK506, ), rapamycin (sirolimus, Tofacitinib Other FK-506 immunosuppressants, and mycophenolate mofetil, such as ethyl mycophenolate mofetil. (d) Antihistamines (H1-histamine antagonists), such as brombutinamide, clofenac, dextrochlorpheniramine, triprolidine, clomastine, diphenhydramine, diphenhydramine, triprolidine, hydroxyzine, methylpyrrolizine, promethazine, trimethoprim, piperhein, cyproheptadine, phenacetin, pheniramine pyramine, astemizole, terfenadine, loratadine, cetirizine, fexofenadine, decarboethoxyloratadine, etc.; (e) Nonsteroidal anti-asthmatic agents (e.g., terbutaline, orsinol, fenoterol, isotretinoin, salbutamol, etc.). (f) Toltero and pirbuterol), theophylline, sodium cromoglycate, atropine, ipratropium bromide, leukotriene antagonists (e.g., zafmlukast, montelukast, prancetta, irastrasil, pobibist, and SKB-106, 203), leukotriene biosynthesis inhibitors (ziluton, BAY-1005); (f) nonsteroidal anti-inflammatory drugs (NSAIDs), such as propionic acid derivatives (e.g., aminolofen, benzoxalofen, buccolic acid, carprofen, fenbufen, fenofen, fluprofen, fluorobiphenyl). Propionic acid, isobutylphenylpropionic acid, indoprofen, ketoprofen, niroprofen, naproxen, oxaprazin, pyrrolifene, pranoprofen, sulprofen, thiaprofen acid, and thioprofen), acetic acid derivatives (e.g., indomethacin, acemetacin, alclofenac, cyclochloroinic acid, diclofenac, fenclofenac, fenclofenac, fenclofenac, furofenic acid, isobutylfenac, isochoric acid, oxpinac, sulinac, thiophenic acid, tometidine, zidomexin, and zolemic acid), fenamic acid derivatives (e.g., fenamic acid derivatives). (g) Flufenamic acid, meclofenamic acid, niflunic acid, and tofenamic acid; biphenyl carboxylic acid derivatives (e.g., diflunisal and flubensal), oxacins (e.g., isoxicam, piroxicam, sudoxicam, and tenoxicam), salicylates (e.g., acetylsalicylic acid and sulfasalazine), and pyrazolone esters (e.g., azaprozin, bezpiperylon, phenprozin, mofiboxin, hydroxybutazine, and phenylbutazone); and cyclooxygenase-2 (COX-2) inhibitors, such as celecoxib. and roficoxib (h) Inhibitors of phosphodiesterase type IV (PDEIV); (i) Gold compounds, such as auronoxine and auroxodil; (j) Etanercept. (k) Antibody therapies, such as octoclopramide (OKT3) and dazumab. Baliximab Inflixi Adalimumab Galimab Rituximab Tocilizumab (l) Other antagonists of chemokine receptors (especially CCR5, CXCR2, CXCR3, CCR2, CCR3, CCR4, CCR7, CX3CR1, and CXCR6); (m) Lubricants or softeners, such as petrolatum and lanolin; (n) keratolytic agents (e.g., tazolotine); (o) Vitamin D3 derivatives, such as calcipotriene or calcipotriol. (p)PUVA; (q)Dianthracene (r) etretinate And isotretinoin; and(s) multiple sclerosis treatment agents, such as interferon beta-1β Interferon (β-1α) Imidazopurine Grammer acetate Glucocorticoids (e.g., prednisolone) and cyclophosphamide; (t) DMARDs, such as methotrexate and leflunomide; (u) other compounds, such as 5-aminosalicylic acid and its prodrugs; hydroxychloroquine; D-penicillamine; antimetabolites, such as imidazoline, 6-mercaptopurine, and methotrexate; DNA synthesis inhibitors, such as hydroxyurea; microtubule disruptors, such as colchicine; and proteasome inhibitors, such as bortezomib. (v) Antibodies against CTLA-4, PD1, or PD-L1. The weight ratio of the compound of the present invention to the second active ingredient can be varied depending on the effective dose of each ingredient. Generally, the effective dose of each ingredient is 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 is typically 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 are also generally within the above range, but in each case, the effective dose of each active ingredient should be used.

[0201] In some embodiments, a method is provided for treating CXCR2 and / or CCR6-mediated diseases or conditions in subjects in need, the method comprising administering an effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of the present disclosure to the subject. In some embodiments, the disease or condition is an acute or chronic inflammatory condition. In some embodiments, the acute or chronic inflammatory condition is psoriasis, dry eye disease, atherosclerosis, discoid lupus erythematosus, rheumatoid arthritis, lupus, radiation-induced fibrotic lung disease, autoimmune bullous skin disease (AIBD), chronic obstructive pulmonary disease, or ozone-induced airway inflammation. In some embodiments, the acute or chronic inflammation is psoriasis.

[0202] In some implementations, the disease is cancer. In some implementations, the cancer is selected from the group consisting of: cutaneous T-cell lymphoma, non-Hodgkin's lymphoma, mycosis fungoides, Paget's reticulosis, Cezary syndrome, granulomatous skin laxity, lymphomatoid papulosis, chronic lichenoid pityriasis, acute lichen pustulosis, CD30+ cutaneous T-cell lymphoma, secondary cutaneous CD30+ large cell lymphoma, non-mycosis fungoides CD30 cutaneous large T-cell lymphoma, pleomorphic T-cell lymphoma, Lennatet's lymphoma, subcutaneous T-cell lymphoma, angiocentric lymphoma, and blastic NK-cell lymphoma. Lymphoma, B-cell lymphoma, Hodgkin's lymphoma (HL), head and neck tumors; squamous cell carcinoma, rhabdomyosarcoma, Lewis lung cancer (LLC), non-small cell lung cancer, esophageal squamous cell carcinoma, esophageal adenocarcinoma, renal cell carcinoma (RCC), colorectal cancer (CRC), acute myeloid leukemia (AML), breast cancer, gastric cancer, small cell neuroendocrine prostate carcinoma (SCNC), liver cancer, glioblastoma, liver cancer, oral squamous cell carcinoma, pancreatic cancer, papillary thyroid carcinoma, intrahepatic cholangiocarcinoma, hepatocellular carcinoma, bone cancer, metastatic and nasopharyngeal carcinoma. In some embodiments, the disease is colorectal cancer. In some embodiments, the disease is cutaneous T-cell lymphoma.

[0203] In some embodiments, the compound is used alone or in combination with one or more other anticancer therapies. In some embodiments, the compound is used in combination with one or more of cytotoxic chemotherapy, anticancer vaccines, antitumor vaccines, anti-immunocytokine therapy, immunocytokine therapy, checkpoint inhibitors and chimeric antigen receptor (CAR) T-cell receptors, and gene transfer therapy. In some embodiments, the compound is used in combination with at least one checkpoint inhibitor. In some embodiments, the compound is used in combination with one or more compounds that block the activity of CTLA-4 (CD152), PD-1 (CD279), PDL-1 (CD274), TIM-3, LAG-3 (CD223), VISTA, KIR, NKG2A, BTLA, PD-1H, TIGIT, CD96, 4-1BB (CD137), 4-1BBL (CD137L), GARP, CSF-1R, A2AR, CD73, CD47, tryptophan 2,3-dioxygenase (TDO), or indoleamine 2,3-dioxygenase (IDO). In some embodiments, the compound is used in combination with one or more agonists of OX40, GITR, 4-1BB, ICOS, STING, or CD40.

[0204] In some embodiments, the compounds of the present invention or their pharmaceutically acceptable salts and / or prodrugs or compositions of the present invention are administered to treat colorectal cancer, metastatic, advanced cutaneous T-cell lymphoma, pancreatic cancer, non-Hodgkin lymphoma, mycosis fungoides, Paget's reticulocytosis, Cezary syndrome, granulomatous skin laxity, lymphomatoid papulosis, chronic lichenoid pityriasis, acute lichen pustulosis, CD30+ cutaneous T-cell lymphoma, secondary cutaneous CD30+ large cell lymphoma, non-mycosis fungoides CD30- cutaneous large T-cell lymphoma, polymorphic T-cell lymphoma, Lennart lymphoma, subcutaneous T-cell lymphoma, angiocentric lymphoma, blastic NK-cell lymphoma, B-cell lymphoma, Hodgkin lymphoma (HL), dry eye disease, atherosclerosis, or discoid lupus erythematosus.

[0205] In some embodiments, the compounds of the present invention or their pharmaceutically acceptable salts and / or their prodrugs or compositions disclosed herein are administered to treat insulin-dependent diabetes mellitus, cystitis, islet cell transplant rejection; kidney transplant rejection; liver transplant rejection; lung transplant rejection, COPD, or influenza.

[0206] In some embodiments, the compounds of the present invention or their pharmaceutically acceptable salts and / or their prodrugs or compositions of the present invention are administered to treat melanoma, glioblastoma, esophageal tumors, nasopharyngeal carcinoma, uveal melanoma, lymphoma, lymphocytic lymphoma, primary CNS lymphoma, T-cell lymphoma, diffuse large B-cell lymphoma, primary mediastinal large B-cell lymphoma, prostate cancer, castration-resistant prostate cancer, chronic myeloid leukemia, Kaposi's sarcoma, fibrosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, angiosarcoma, lymphangiosarcoma, synovial tumors. Tumors, meningiomas, leiomyosarcomas, rhabdomyosarcomas, soft tissue sarcomas, sarcomas, sepsis, bile duct tumors, basal cell carcinoma, thymoma, thyroid cancer, parathyroid carcinoma, uterine cancer, adrenal cancer, liver infection, Merkel cell carcinoma, neurotumors, follicular center lymphoma, colon cancer, Hodgkin's disease, non-Hodgkin's lymphoma, leukemia, chronic or acute leukemia (including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia), multiple myeloma, ovarian tumors, myelodysplastic syndromes, skin diseases. Malignant melanoma of the skin or eye, renal cell carcinoma, small cell lung cancer, lung cancer, mesothelioma, breast cancer, squamous non-small cell lung cancer (SCLC), non-squamous NSCLC, colorectal cancer, ovarian cancer, gastric cancer, hepatocellular carcinoma, pancreatic cancer, pancreatic duct adenocarcinoma, head and neck squamous cell carcinoma, head and neck cancer, gastrointestinal cancer, stomach cancer, bone cancer, skin cancer, rectal cancer, anal cancer, testicular cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, esophageal cancer, small intestine cancer, endocrine system cancers, urethral cancer, penile cancer, bladder cancer, kidney cancer, ureteral cancer Renal pelvis cancer, central nervous system (CNS) tumors, tumor angiogenesis, spinal tumors, brainstem gliomas, pituitary adenomas, epidermoid carcinomas, asbestosis, carcinomas, adenocarcinomas, papillary carcinomas, cystadenocarcinomas, bronchial carcinomas, renal cell carcinomas, transitional cell carcinomas, choriocarcinomas, seminomas, embryonal carcinomas, Wilms' tumors, pleomorphic adenomas, hepatocellular papillary tumors, renal tubular adenomas, cystadenomas, papillomas, adenomas, leiomyomas, rhabdomyomas, hemangiomas, lymphangiomas, osteomas, chondromas, lipomas, and / or fibromas.

[0207] combination therapy

[0208] The compounds of this invention can be supplied alone or in combination with one or more other drugs. Possible combination partners may include additional anti-angiogenic factors and / or chemotherapeutic agents (e.g., cytotoxic agents) or radiation, cancer vaccines, immunomodulators, checkpoint inhibitors, anti-angiogenic agents, signal transduction inhibitors, antiproliferative agents, apoptosis inducers, alkylating agents, nitrosoureas, antimetabolites, anticancer antibiotics, plant-derived alkaloids, topoisomerase inhibitors, hormonal drugs, hormone antagonists, aromatase inhibitors, P-glycoprotein inhibitors, platinum complex derivatives, antifibrotic agents, radiotherapy, radiotherapy agents, and gene expression regulators.

[0209] Examples of other therapeutic agents (administered alone or in the same pharmaceutical composition) that can be used in combination with the compounds or compositions of the present invention include, but are not limited to, modifiers of CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCR10, CCR11, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, CXCR7, CX3CR1, ChemR23, C5aR, C5a and C5, or any combination thereof. In some embodiments, the modifier is an antagonist.

[0210] Examples of other therapeutic agents (administered alone or in the same pharmaceutical composition) that can be combined with the compounds or compositions of the present invention include, but are not limited to: therapeutic antibodies, bispecific antibodies, and "antibody-like" therapeutic proteins (such as...). Fab derivatives), antibody-drug conjugates (ADCs), viruses, oncolytic viruses, gene-modifying agents or editing agents such as CRISPR (including CRISPRCas9), zinc finger nucleases or synthetic nucleases (TALEN), CAR (chimeric antigen receptor) T-cell immunotherapeutic agents, cytokines, vaccines, vaccine adjuvants, GM-CSF, M-CSF, G-CSF, interferon-α, β or γ, IL-1, IL-2, IL-3, IL-12, poly(I:C), CPG, cyclophosphamide, cyclophosphamide analogs, anti-TGF and imatinib (Gleevac), mitotic inhibitors, kinase inhibitors, paclitaxel, sunitinib (Sutent), anti-angiogenic agents, aromatase inhibitors Agents, letrozole, A2a adenosine receptor (A2AR) antagonists, adenosine receptor modulators, A3 adenosine receptor modulators, angiogenesis inhibitors, anthracyclines, oxaliplatin, doxorubicin, TLR4 antagonists, IL-18 antagonists, Btk tyrosine kinase inhibitors, Erbb2 tyrosine kinase receptor inhibitors; Erbb4 tyrosine kinase receptor inhibitors, mTOR inhibitors, thymidine synthase inhibitors, EGFR tyrosine kinase receptor inhibitors, epidermal growth factor antagonists, Fyn tyrosine kinase inhibitors, Kit tyrosine kinase inhibitors, Lyn tyrosine kinase inhibitors, NK cell receptor modulators, PDGF receptor antagonists, PARP inhibitors, poly-ADP ribose polymerase inhibitors, poly-ADP Ribose polymerase 1 inhibitors, poly-ADP-ribose polymerase 2 inhibitors, poly-ADP-ribose polymerase 3 inhibitors, galactosyltransferase modulators, dihydropyrimidine dehydrogenase inhibitors, orotate phosphoribosyltransferase inhibitors, telomerase modulators, mucin 1 inhibitors, mucin inhibitors, secretin agonists, TNF-related apoptosis-inducing ligand modulators, IL-17 gene stimulators, interleukin-17E ligands, neurokinin receptor agonists, cyclin G1 inhibitors, checkpoint inhibitors, PD-1 inhibitors, PD-L1 inhibitors, CTLA4 inhibitors, topoisomerase I inhibitors, Alk-5 protein kinase inhibitors, connective tissue growth factor ligand inhibitors, Notch-2 receptor antagonists, Notc h-3 receptor antagonists, hyaluronidase stimulants, MEK-1 protein kinase inhibitors, phosphoinositol-3 kinase inhibitors, MEK-2 protein kinase inhibitors, GM-CSF receptor modulators; TNFα ligand modulators, mesothelin modulators, asparaginase stimulants, CSF2 gene stimulants, caspase-3 stimulants; caspase-9 stimulants, PKN3 gene inhibitors, hedgehog protein inhibitors, smooth receptor antagonists, AKT1 gene inhibitors, DHFR inhibitors, thymidine kinase stimulants, CD29 modulators, fibronectin modulators, interleukin-2 ligands, serine protease inhibitors, D40LG gene stimulants; TNFSF9 gene stimulants, 2-ketoglutarate dehydrogenase inhibitors.TGF-β type II receptor antagonists, Erbb3 tyrosine kinase receptor inhibitors, cholecystokinin (CCK2) receptor antagonists, Wilms tumor protein modulators, RasGTPase modulators, histone deacetylase inhibitors, RafB protein kinase inhibitors, cyclin-dependent kinase 4 inhibitors, estrogen receptor β modulators, 4-1BB inhibitors, 4-1BBL inhibitors, PD-L2 inhibitors, B7-H3 inhibitors, B7-H4 inhibitors, BTLA inhibitors, HVEM inhibitors, TIM3 inhibitors, TIGIT inhibitors, NKG2A inhibitors, GAL9 inhibitors, LAG3 inhibitors, PD-1H inhibitors, PD96 inhibitors, VISTA Inhibitors, KIR inhibitors, 2B4 inhibitors, CD160 inhibitors, CD66e modulators, angiotensin II receptor antagonists, connective tissue growth factor ligand inhibitors, Jak1 tyrosine kinase inhibitors, Jak2 tyrosine kinase inhibitors, dual Jak1 / Jak2 tyrosine kinase inhibitors, angiotensin-converting enzyme 2 stimulators, growth hormone receptor antagonists, galactoglobulin-3 inhibitors, checkpoint kinase 2 modulators, sodium-glucose transporter-2 inhibitors, endothelin ET-A antagonists, mineralocorticoid receptor antagonists, endothelin ET-B antagonists, higher glycosylation product receptor antagonists, adrenocorticotropic hormone ligands, farnesoid X receptor agonists, G proteins Coupled with bile acid receptor 1 agonists, aldose reductase inhibitors, xanthine oxidase inhibitors, PPARγ agonists, prostaglandin receptor antagonists, FGF receptor antagonists, PDGF receptor antagonists, TGFβ antagonists, P3 protein modulators, p38MAP kinase inhibitors, VEGF-1 receptor antagonists, protein tyrosine phosphatase β inhibitors, Tek tyrosine kinase receptor stimulators, PDE5 inhibitors, mineralocorticoid receptor antagonists, ACE inhibitors, I-κB kinase inhibitors, NFE2L2 gene stimulators, nuclear factor κB inhibitors, STAT3 gene inhibitors, NADPH oxidase 1 inhibitors, NADPH oxidase 4 inhibitors, PDE4 inhibitors, renin inhibitors, and FURI. N gene inhibitors, MEKK-5 protein kinase inhibitors, membrane copper amine oxidase inhibitors, integrin α-V / β-3 antagonists, insulin sensitizers, kallikrein 1 modulators, cyclooxygenase inhibitors, complement C3 modulators, microtubule binding agents, macrophage mannose receptor 1 modulators, phenylalanine hydroxylase stimulators, diphenhydramine, bexarotine, vorinostat, romidesin, pralatrexate, prednisone, prednisolone, CCX354, CCX9588, CCX140, CCX872, CCX598, CCX6239, CCX9664, CCX2553, CCX2991, CCX282, CCX025, CCX507, CCX430, CCX765.CCX224, CCX662, CCX650, CCX832, CCX168, CCX168-M1, Bavitimab, IMM-101, CAP1-6D, Rexin-G, Genistein, CVac, MM-D37K, PCI-27483, TG-01, Mocetinostat, LOAd-703, CPI-613, Upamostat, CRS-207, NovaCaps, Trametinib, Atu-027, Sonidegib, GRASPA, Trabedsen Abedersen, Nastorazepide, Dendritic Cell Immunotherapy (Vaccell), Ogovovomab, Istiratumab, Refametinib, Regrafenib, Lapatinib, Selmetinib, Recaprapab, Pelareorep, Tarextumab, Pelylated Hyaluronidase, Varlitinib, Aglatimagenebesadenovec, GBS-01, GI-4000, WF-10, Galuniser tib), afatinib, RX-0201, FG-3019, pertuzumab, DCVax-Direct, selinexor, glucosamine, virulencerin, yttrium (90Y) civatuumab, clivatuzumab, brovudine, nimotuzumab, algenpantucel-L, tegafur + gemmester + oxazine potassium + leucovorin, olaparib, ibrutinib, pirarubicin, Rh-Apo2L, tertomotide, tegafur + gemmester + oteracis potassium, tegafur + gemmester + oteracis potassium, masitinib nib), Lexycin-G, Mitomycin, Erlotinib, Doxorubicin, Dexamethasone, Vincristine, Cyclophosphamide, Fluorouracil, Topotecan, Paclitaxel, Interferon, Platinum Derivatives, Taxane, Paclitaxel, Vincristine, Anthracyclines, Doxorubicin, Epipodophyllotoxin, Etoposide, Cisplatin, Rapamycin, Methotrexate, Actinomycin D, Dolastati 10, Colchicine, Emetine, Trimethoprim, Chlorfenapyridine, Cyclosporine, Daunorubicin, Teniposide, Amphotericin B, Alkylating Agents, Chlorobutazone, 5-Fluorouracil, Camptothecin, Cisplatin, Metronidazole, Gleevec, Avastin, Panitumumab, AbaricillinInterleukin, alenzab, retinoic acid, allopurinol, hexamethylmelamine, amifostine, anastrozole, arsenic trioxide, asparaginase, azacitidine, AZD9291, live BCG vaccine, bevacizumab, fluorouracil, besalodin, bleomycin, bortezomib, busulfan, capprotestone, capecitabine, camptothecin, carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cladribine, clofarabine, cyclophosphamide, cytarabine, daunorubicin, afadabetine, daunorubicin, denileukin, dexrazosen, docetaxel, doxorubicin (neutral), doxorubicin hydrochloride, drotaldoxin propionate, epirubicin, epibasine, etoposide, etoposide phosphate, etoposide, etoposide Simestane, Filgrastim, Fluorouracil Fludarabine, Fulvestrant, Gefitinib, Gemcitabine, Gemtuzumab, Goserelin Acetate, Histamine Relin Acetate, Hydroxyurea, Teimimobazide, Idarubicin, Ifosfamide, Imatinib Mesylate, Interferon Alpha-2a, Interferon Alpha-2b, Irinotecan, Lenalidomide, Letrozole, Calcium Leucovorin, Leuprorelin Acetate, Levamisole, Lomustine, Medroxyprogesterone Acetate, Melphalan, Mercaptopurine, 6-MP, Mesna, Methotrexate, Methoxam, Mitomycin C, Mitotan, Mitoxantrone, Nandrolone, Nerapine, Nofetumomab, Olepiridone Interleukin, Oxaliplatin, Albumin-Bound Paclitaxel, Palivmin, Pamidronate Disodium, Pergamase, Aspartate Aminoaminase, Pefepristone, Pemetrexed disodium, Pentostatin, Piperbromide, Propofol, Porphyrin, Procarbazine, Adipine, Raburicase, Rituximab, Lociletinib, Saxaglastine, Sorafenib, Streptozotocin, Sunitinib Maleate, Talc, Tamoxifen, Temozolomide, Teniposide, VM-26, Testrolide, Thioguanine, 6-TG, Thiotepa, Topotecan, Toremifene, Tosimomab, Trastuzumab, Retinoic Acid, ATRA, Uracil Mustard, Penrubicin, Vincristine, Vincristine, Vinorelbine, Zoledronic Acid, Zoledronic Acid, Pembrolizumab, Nivolumab, IBI-308, mDX-400, BGB-108, MEDI-0680, SH R-1210, PF-06801591, PDR-001, GB-226, STI-1110, Dvalumab, Atezolizumab, Acitumab, BMS-936559, ALN-PDL, TSR-042, KD-033, CA-170, STI-1014, FOLFIRINOX, KY-1003, Olmesartan Medoxomil, Candesartan, PBI-4050, Baricitinib, GSK-2586881, Losartan, Dapagliflozin Propylene Glycol, Pevisomylene, GR-MD-02, Canagliflozin, Irbesartan, FG-3019, Atrasentan, Finerenone, SparsentanBosentan, defibrinogen polynucleotide, femasartan, azeliragon, pyridoxine, adrenocorticotropic hormone, INT-767, epalrestat, topirostat, SER-150-DN, pirfenidone, VEGFR-1mAb, AKB-9778, PF-489791, SHP-627, CS-3150, imidapril, perindopril, captopril, enalapril, lisinopril, zolfenpril, lisinopril, quinapril, benazepril, trandopril, cilazapril, fosinopril, ramipril, methylbardosolon, irbesartan + hyperlipidemia, GKT-831, MT-3995, TAK-648, TAK-272, GS-4997. DW-1029M, ASP-8232, VPI-2690B, DM-199, rhein, PHN-033, GLY-230, sapropterin, sulodexix, lirilumab, IPH-4102, IPH-2101, IMP-321, BMS-986016, MGD-013, LAG-525, durvalumab, monalizumab, MCLA-134, MBG-453, CA-170, AUPM-170, AUPM-327, resminostat, ipilimumab, B GB-A317, tremelimumab, REGN-2810, AZD-5069, masitinib, binimetinib, trametinib, ruxolitinib, dabrafenib, linaclotide, ipilimumab, apatinib, nintedanib, cabozantinib, pazopanib, belinostat, panitumumab, guardixetine Guadecitabine, vismodegib, vemurafenib, dasatinib, trimemumab, bevacizumab, oxaliplatin, aflibercept, vandetanib, everolimus, thalidomide, veliparib, encorafenib, napabucasin, alpelisib, and axitinib.Cedilanib, Necitumumab, Ramucirumab, Irofulven, Trifluridine + Tipiracil, Donafenib, Pacritinib, Pexastimogene devacirepvec, tivantinib, GNR-011, talaporfin, piclidenoson, decitabine, ganitumab, panobinostat, rintatolimod, polmacoxib, levofolinate, famitinib, votumumab, tivozanib, entinostat, plitidepsin, lefitolimod, OSE-2101, and vetivantinib. Vitespen, TroVax, bromocriptine, midostaurin, fosbretabulin, fruquintinib, ganetspib, brivanib, anlotinib, L19-TNF-α, racotumomab, Novaferon, raltitrexed, enzastaurin, GM-CT-01, acitumomab, denosumab, bexarotin, vorinostat, romidesin, pralatrexate, prednisone, prednisolone, or any combination thereof.

[0211] Examples of other therapeutic agents (administered alone or in the same pharmaceutical composition) that can be combined with the compounds or compositions of the present invention include, but are not limited to: MP-1032, secukinumab, betamethasone, cyclosporine, certolizumab, and pegylated certolizumab. pegol), VTP-43742, bimekizumab, GSK-2982772, amiselimod, KD-025, ustekinumab, etanercept, guselkumab, apremilast, dimethyl fumarate + calcium monoethyl fumarate + magnesium monoethyl fumarate - zinc monoethyl fumarate, infliximab, risankizumab, ixekizumab, mometasone, brodamarumab, adalimumab, tofacitinib, olopatadine, tazarotene, dimethyl fumarate, Trichuris suis oocystsOVA), BTT-1023, voclosporin, seletalisib, INV-103, piclidenoson, GR-MD-02, PRX-167700, LYC-30937EC, namilumab, LY-3074828, LEO-32731, acitretin, calcipotriol, WBI-1001, clobetasol propionate, betamethasone, ZPL-389, bertilimumab, AKP-11, ZPL-521, crisaborole, CLS-0 08, IMO-8400, bleselumab, calcipotriol, tildrakizumab, KX-01, 18C3, DSXS-1411, DLX-105, remetinostat, prurisol, S-414114, GLG-801, inecalcitol, masalcitol + betamethasone, TAB-08, alefacept, ubetasol, toreforant, calcipotriol, betamethasone dipropionate, tregalizumab CJM-112, Neihulizumab, Betamethasone Valerate, P-3072, P-3073, Methotrexate, GSK2981278A, Calcipotriol + Betamethasone Dipropionate, LEO-124249, AVX-001, Calcipotriol + Betamethasone Dipropionate, Dimethyl Fumarate, Halometasol Propionate + Tazarotene, Calcipotriol, Calcipotriol + Betamethasone, Arivir A, DFD-06, Bengal Rose Sodium, C-82, TU-2100, CT-327, Paricalcitol, Fluocinolone Acetate, Clobetasol Propionate + Retinoic Acid, GK-664-S, Tazarotene + Betamethasone, Irimethasone Itolizumab, betamethasone valerate, IMO-3100, PUR-0110, LEO-29102, orilotimod, maxacalcitol, IR-502, myristyl nicotinate, aganirsen, methotrexate, mometasone furoate, BCG polysaccharide + nucleic acid injection, lithium succinate, orilotimod, LAS-41004, calcitriol, GMDP, mometasone furoate, MOL-4249, aminopterin, tacacool, dithranol, halometasone, anapos, osimertinib, and AGEN-1884.

[0212] reagent kits and packaging

[0213] The terms "kit" and "pharmaceutical kit" refer to commercial kits or packages that contain one or more pharmaceutical compositions and instructions for use in one or more suitable containers. In one embodiment, a kit containing a compound of formula (A), (A1), (A2), (I), or (Ia1) or a pharmaceutically acceptable salt thereof, along with instructions for administration, is provided. In another embodiment, a kit containing compounds of formulas (A), (A1), (A2), (I), and (Ia1) or a pharmaceutically acceptable salt thereof, along with one or more additional therapeutic agents (e.g., one, two, three, one or two, or one to three), along with instructions for administration, is provided.

[0214] In one embodiment, the compound of the invention is formulated into a dosing unit packaged in a single package. The single package includes, but is not limited to, bottles, child-safe bottles, ampoules, and tubes. In one embodiment, the compound of the invention and optionally additional therapeutic agents are formulated into a dosing unit, and each single dosing unit is packaged separately in a single package. Such individually packaged units may contain any form of pharmaceutical composition, including but not limited to liquid, solid, powder, granule, effervescent powder or tablet, hard or soft capsule, emulsion, suspension, syrup, suppository, tablet, lozenge, diamond-shaped lozenge, solution, oral patch, film, oral gel, chewable tablet, chewing gum, and disposable syringe. Such individually packaged units may be combined in a package made of one or more types of paper, cardboard, paperboard, metal foil, and plastic foil, such as blister packs. One or more dosing units may be administered once or several times daily. One or more dosing units may be taken three times a day. One or more dosing units may be administered twice a day. One or more dosing units may be administered on the first day, and one or more dosing units may be administered on subsequent days.

[0215] General synthesis procedure

[0216] The implementation plan also relates to methods and intermediates that can be used to prepare the subject compound or its pharmaceutically acceptable salts.

[0217] Exemplary chemical entities that can be used in implementing the methods will now be described with reference to the illustrative synthetic schemes of the general preparation described herein and the following specific examples. Those skilled in the art will recognize that, in order to obtain the various compounds described herein, starting materials can be suitably selected such that the final desired substitution will be carried out by the reaction scheme, with or without protection, to produce the desired product. Alternatively, at the position of the final desired substituent, suitable groups may be required or desired, which can be carried out by the reaction scheme and suitably replaced with the desired substituent. Furthermore, those skilled in the art will recognize that the transformations shown in the schemes below can be performed in any order compatible with the function of the particular side groups.

[0218] Representative syntheses of the compounds disclosed herein are described in the following schemes and specific examples. Schemes 1 and 2 are provided as further embodiments of the present disclosure, and a general method for preparing the compounds of the present disclosure, including compounds of formulas (A), (A1), (A2), (I), and (Ia1), is illustrated, and this method can be used to prepare additional compounds of formulas (A), (A1), (A2), (I), and (Ia1). This method is compatible with various functionalities.

[0219] Option 1

[0220]

[0221] The amino group of B1 can react with 3,4-dimethoxycyclobut-3-ene-1,2-dione to give B2. B2 can then react with the amino group of B3 to provide B4.

[0222] Option 2

[0223]

[0224] B7 can be obtained by reducing the cyano group in B6, for example by hydrogenation, followed by cyclization. Alternatively, B5 (where X represents a leaving group, such as a halogen or toluenesulfonate, and where R is an alkyl group) can react with NH3 to form the cyclized product B7. B7 can be reacted with HNO3 to introduce a nitro group in the presence of an acid such as sulfuric acid, yielding B8. B9 can be obtained by, for example, hydrogenation followed by reduction of the nitro group in B8. Example

[0225] The following embodiments are provided to illustrate, but do not limit, the claimed invention.

[0226] The reagents and solvents used can be obtained from commercial sources such as Aldridge Chemical Co., Ltd. (Milwaukee, Wisconsin, USA). Recorded on a Varian Mercury 400MHz NMR spectrometer. 1¹H-NMR. Peaks are presented significantly relative to TMS and listed in the following order: multiplicity (s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet) and number of protons. Mass spectrometry results are reported as mass-to-charge ratios, followed by the relative abundance of each ion (as an interpolation component). In the table, individual m / e values ​​for M+H (or, as recorded, MH) ions containing the most common atomic isotopes are reported. Isotopic patterns correspond to the expected formulas in all cases. Electrospray ionization (ESI) mass spectrometry analysis was performed on a Hewlett-Packard MSD ESI mass spectrometer using an HP1100 HPLC equipped with an Agilent ZORBAX column (SB-C18, 2.1 x 50 mm, 5 μm column) for sample delivery. Typically, the analyte was dissolved in methanol at 0.1 mg / mL, and 1 μL was injected into the mass spectrometer along with the delivery solvent, which scanned from 100 to 1500 Daltons. All compounds can be analyzed in ESI positive ion mode using acetonitrile / water containing 1% formic acid as the delivery solvent. The compounds listed below can also be analyzed in ESI negative ion mode using acetonitrile / water with 2 mM NH4OAc as the delivery system.

[0227] The following abbreviations are used in the embodiments and throughout the description of the invention:

[0228] HPLC, high-performance liquid chromatography; DMF, dimethylformamide; TFA, trifluoroacetic acid; THF, tetrahydrofuran; EtOAc, ethyl acetate; BOC2O, di-tert-butyl dicarbonate or BOC anhydride; HPLC, high-performance liquid chromatography; DIPEA, diisopropylethylamine; HBTU, O-(benzotriazol-1-yl)-N,N,N,N-tetramethylurea hexafluorophosphate; dppf, 1,1'-bis(diphenylphosphine)ferrocene; Pd2(dba)3, tris(dibenzylideneacetone)dipalladium(O); DIPEA, diisopropylethylamine; DMP, dimethyl phthalate; Me, methyl; Et, ethyl; DCM, dichloromethane.

[0229] Compounds within the scope of this invention can be synthesized using various reactions known to those skilled in the art. Those skilled in the art will also recognize that alternative methods can be employed to synthesize the target compounds of this invention, and that the methods described herein are not exhaustive, but do provide a broad and practical approach to the synthesis of the target compounds.

[0230] Certain molecules claimed in this patent may exist in different enantiomeric and diastereomeric forms, and all such variants of these compounds are claimed.

[0231] In this paper, the detailed description of the experimental steps used to synthesize key compounds leads to the description of molecules by physical data that identify them and structural descriptions associated with them.

[0232] Those skilled in the art will also recognize that acids and bases are frequently used in standard post-processing in organic chemistry. In the experimental steps described in this patent, salts of the parent compound may sometimes be generated if they possess the necessary intrinsic acidity or basicity.

[0233] Example 1: Synthesis of 3-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]-4-ethoxy-cyclobut-3-ene-1,2-dione

[0234]

[0235] Step a: In a 5L three-necked round-bottom flask equipped with a mechanical stirrer, (R)-2-methylpropane-2-sulfinamide (100g, 0.825mol), 2,2-dimethylpropanal (78.2g, 0.907mol), and tetraethoxytitanium (414.1g, 1.815mol) were added to dichloromethane (1.5L). The reaction mixture was stirred at room temperature for 12 hours, then sodium sulfate decahydrate (260g) was added, followed by diatomaceous earth (500g). The mixture was stirred at room temperature for 5 hours, filtered through diatomaceous earth, and washed with dichloromethane (1L). The filtrate was concentrated under vacuum and dried under vacuum overnight to give a brown oily [N(E),S(R)]-N-(2,2-dimethylpropane)-2-methyl-propane-2-sulfinamide (150g, 96%), which was used for the next step without further purification. 1 HNMR (400MHz, CD3OD) δ7.80 (s, 1H), 1.10 (s, 9H), 1.08 (s, 9H); MS: C 19 H 19 The calculated value of NOS (ES) m / z is 190.1, and the measured value is also 190.1.

[0236] Step b: Add 2-methylfuran (31.1 mL, 345.1 mmol, 1.5 equivalents) and anhydrous Et₂O (300 mL) to a 1 L three-necked flask equipped with a feeding funnel, and then cool in an ice bath. Add n-BuLi (2.5 M, 120 mL, 299 mmol, 1.3 equivalents) dropwise in hexane over approximately 35 minutes. Stir the mixture at 0 °C for 30 minutes, then at room temperature for 40 minutes, and then cool again to 0 °C. Add solid MgBr₂xEt₂O (77.2 g, 299.1 mmol, 1.3 equivalents) and stir the mixture at 0 °C for 30 minutes, then at room temperature for 20 minutes.

[0237] In a 5L three-necked flask equipped with a mechanical stirrer and an internal thermometer, the imine (43.5 g, 230.1 mmol) from step a was dissolved in anhydrous toluene (1.2 L) and cooled to an internal temperature of -70 °C. The lithium salt solution from the previous section was added over 56 minutes, maintaining the internal temperature between -70 and -67.8 °C. After the addition was complete, the reaction mixture was stirred at -70 °C for 1 hour, then overnight at room temperature. The reaction mixture was slowly quenched with saturated NH4Cl solution (400 mL) and water (400 mL), then stirred at room temperature for 15 minutes. The organic layer was separated and washed with brine (200 mL). The combined aqueous layers were extracted with ethyl acetate (300 mL). The organic phase was dried over MgSO4, filtered, and evaporated to give an orange oil. The crude product was dissolved in hexane (500 mL) and allowed to crystallize overnight at -20 °C to give a yellow solid. The solid was filtered, the mother liquor was evaporated, and crystallized again from hexane (50 mL) to give the product (51.9 g, 83%), which was a pure diastereomer. MS: C 14 H 26 NO2S[M+H] + The calculated value of (ES)m / z is 272.2, and the measured value is also 272.2.

[0238] Step c: N-[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]-2-methyl-propane-2-sulfinamide (51.9 g, 191.5 mmol) from the previous step was dissolved in methanol (100 mL) and cooled in an ice bath, then 2 M HCl in diethyl ether (191.5 mL, 383.0 mmol, 2 equivalents) was added. The cooling bath was removed and the reaction mixture was stirred at room temperature for 2 hours. The solvent was removed under vacuum, and anhydrous diethyl ether (300 mL) was added to the residue. The resulting mixture was filtered. Water (100 mL) and 1 M NaOH aqueous solution (200 mL) were added to the solid. The product was extracted with dichloromethane (3 x 100 mL), the combined organic layers were dried over MgSO4, filtered, and evaporated to give a yellow oil (27.2 g, 85%). MS: C 10 H 15 O[(M–NH3)+H] + The calculated value of (ES)m / z is 151.1, and the measured value is 151.1.

[0239] Step d: 3,4-Diethoxycyclobut-3-en-1,2-dione (15.9 g, 93.5 mmol, 1.05 equivalents) was dissolved in anhydrous ethanol (150 mL) and cooled in an ice bath. Then, a solution of (1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propane-1-amine (14.9 g, 89.0 mmol) in anhydrous ethanol (50 mL) was added dropwise, and the reaction mixture was stirred overnight at room temperature. Excess solvent was evaporated, and the residue was stirred with hexane (500 mL) until a solid precipitate formed. The solid was filtered, washed with hexane (100 mL), and dried under high vacuum to give the title compound (24.4 g, 94%). MS: C 16 H 22 NO4[M+H] + The calculated value of (ES)m / z is 292.1, and the measured value is also 292.1.

[0240] Example 2: Synthesis of 2-[4-chloro-7-[[2-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]benzoic acid

[0241]

[0242] Step a: Cool a 4L Erlenmeyer flask containing 100.0g (0.586mole) of concentrated H₂SO₄ (500mL) in an ice bath. Add dropwise 70% HNO₃ (45.2mL, 0.703mole, 1.2 equivalents), stir the reaction mixture at 0°C for 2 hours, then carefully quench with ice and dilute to 4L with cold water. Filter the white solid, wash with water, and dry under high vacuum (127g, quantitative) to give a mixture of 3-chloro-2-methyl-6-nitrobenzoic acid and 3-chloro-2-methyl-5-nitrobenzoic acid in a 3:1 ratio. MS: C₈H₅ClNO₄[MH] - The calculated value of (ES)m / z is 214.0, and the measured value is also 214.0.

[0243] Step b: The isomeric acid mixture from the previous step (50 g, 232.0 mmol) was dissolved in anhydrous DMF (200 mL), and anhydrous Na₂CO₃ (27.0 g, 255.2 mmol, 1.1 equivalents) was added. The reaction mixture was stirred at room temperature for 30 min. Methane iodine (15.9 mL, 255.2 mmol, 1.1 equivalents) was added, and stirring was continued at room temperature for 3 h. The reaction mixture was diluted with water (1.2 L), and the product was extracted with Et₂O (3 × 250 mL). The combined organic layers were washed with brine (4 x 100 mL), dried over MgSO₄, filtered, and evaporated to give a yellow oil (49.7 g, 93%).

[0244] Step c: The mixture of isomeric esters from the previous step (49.7 g, 216.5 mmol) was dissolved in CCl4 (400 mL) and N-bromosuccinimide (57.8 g, 324.7 mmol, 1.5 equivalents) was added, followed by benzoyl peroxide (10.4 g, 43.2 mmol, 0.20 equivalents). The reaction mixture was refluxed and stirred overnight, then cooled to room temperature and filtered. The filtrate was evaporated, and the residue was purified by silica gel chromatography (100:0 to 9:1 hexane:ethyl acetate) to give a yellow monomeric solid (44.1 g, 66%). 1 H NMR (400MHz, CDCl3) δ8.07 (d, J = 9.2 Hz, 1H), 7.65 (d, J = 9.2 Hz, 1H), 4.63 (s, 2H), 4.01 (s, 3H).

[0245] Step d: A suspension of the product from the previous step (616 mg, 2 mmol), methyl anthranilate (302 mg, 2 mmol), and K₂CO₃ (553 mg, 4 mmol) dissolved in anhydrous acetonitrile was heated to 85 °C overnight in a 40 mL sealed reaction flask. The reaction mixture was then cooled to room temperature, diluted with ethyl acetate, and filtered. The filtrate was concentrated to give an uncyclized crude product (800 mg). This crude product was dissolved in acetic acid (5 mL) and heated to 120 °C overnight to give the cyclized product. The cyclized product was diluted with ethyl acetate, washed with water and a saturated aqueous solution of NaHCO₃, dried (Na₂SO₄), filtered, and concentrated. The residue was adsorbed onto silica and purified by silica gel chromatography (0-50% ethyl acetate in hexane) to give the desired product (350 mg, 50%). MS: C 16 H 11 ClN2O[M+H] + The calculated value of (ES)m / z is 347.0, and the measured value is also 347.0.

[0246] Step e: Iron powder (224 mg, 4 mmol) was added to a stirred mixture of methyl 2-(4-chloro-7-nitro-1-oxo-isoindoline-2-yl)benzoate (347 mg, 1 mmol) in ethanol at room temperature, followed by the addition of a 4 M HCl solution in dioxane (2 mL, 8 mmol). The reaction mixture was stirred at room temperature for 1 hour, then concentrated under vacuum. The residue was diluted with ethyl acetate and neutralized with saturated sodium bicarbonate solution and extracted with ethyl acetate (2 × 5 mL). The combined organic layers were dried (Na₂SO₄), filtered, and concentrated under vacuum. The crude product was purified by silica gel chromatography (0–100% ethyl acetate in hexane) to give methyl 2-(7-amino-4-chloro-1-oxo-isoindoline-2-yl)benzoate (200 mg, 0.63 mmol, 63%) as a yellow powder. MS: C 16 H 13 ClN2O3[M+H] + The calculated value of (ES)m / z is 317.0, and the measured value is also 317.0.

[0247] Step f: A dichloromethane solution of methyl 2-(7-amino-4-chloro-1-oxo-isoindoline-2-yl)benzoate (109 mg, 0.34 mmol) and 3-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]-4-ethoxy-cyclobut-3-ene-1,2-dione (100 mg, 0.34 mmol) was stirred at 0 °C, and a 2 M trimethylaluminum solution in toluene (0.68 mL, 1.36 mmol) was added. The solution was stirred at 0 °C for 1 hour, then warmed to room temperature and stirred for another hour. The reaction mixture was cooled to 0 °C, quenched with 5% hydrochloric acid and diluted with water, and then extracted with ethyl acetate (2 × 5 mL). The organic layer was dried (Na₂SO₄), filtered, and concentrated under vacuum. The crude product was purified by HPLC to obtain methyl 2-[4-chloro-7-[[2-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]benzoate (65 mg, 0.12 mmol, 34%). MS: C 30 H 28 ClN3O6[MH] + The calculated value of (ES)m / z is 560.0, and the measured value is also 560.0.

[0248] Step g: Excess lithium hydroxide was added to a solution of methyl 2-[4-chloro-7-[[2-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]benzoate (56 mg, 0.1 mmol) in tetrahydrofuran (1 mL), methanol (0.1 mL), and water (0.1 mL). The resulting mixture was stirred at room temperature for 1 hour. The reaction was acidified with 5% hydrochloric acid solution and extracted with ethyl acetate. The organic layer was dried (Na₂SO₄), filtered, and concentrated under vacuum. The crude product was purified by HPLC to obtain a yellow solid 2-[4-chloro-7-[[2-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]benzoic acid solid (30 mg, 0.05 mmol, 50%). 1 H NMR(400MHz, DMSO-d6)δ9.89(s,1H),9.06(d,J=10Hz,1H),7.85(dd,J=7.6,1.6Hz,1H),7.65–7.41(m,5H),6.10(d,J =2.6Hz,1H),5.95(d,J=2.6Hz,1H),5.03(d,J=10.4Hz,1H),4.80(dd,J=20,10Hz,2H),2.20(s,3H),0.87(s,9H).MS: C 29 H 26 The calculated (ES) m / z value of ClN3O6[MH]- is 546.0, and the measured value is also 546.0.

[0249] Example 3: 3-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]-4-[(5-fluoro-3-oxo-isoindoline-4-yl)amino]cyclobut-3-ene-1,2-dione

[0250]

[0251] Step a: Add methyl 2-bromo-5-fluorobenzoate (48 g, 206 mmol), cuprous cyanide (37 g, 412 mmol), and DMF (200 mL) to a 500 mL round-bottom flask. Heat the mixture at 110 °C overnight, then cool to room temperature. Add diethyl ether (1.5 L) and diatomaceous earth (100 g), and stir the mixture at room temperature for 30 minutes. Filter the solid, wash the filtrate with brine (3 × 200 mL), and dry with MgSO4. Evaporate the solvent under reduced pressure to give the desired product as a colorless solid (31 g, 84%). MS: C9H7FNO2[M+H]+ The calculated value of (ES)m / z is 180.1, and the measured value is 180.1.

[0252] Step b: 10% Pd-C (1.0 g) was added to a methanol (200 mL) solution of methyl 2-cyano-5-fluorobenzoate (10 g, 56 mmol) at room temperature. The resulting mixture was stirred overnight under a hydrogen (50 psi) atmosphere. The reaction mixture was filtered through diatomaceous earth and the filtrate was concentrated under reduced pressure to give the desired product as a colorless solid (8.0 g, 90%). MS: C8H7FNO[M+H + The calculated value of (ES)m / z is 152, and the measured value is also 152.

[0253] Step c: A pre-cooled mixture of concentrated H₂SO₄ (26 mL) and nitric acid (6 mL) was added dropwise to a suspension of 6-fluoroisoindolin-1-one (8.0 g, 5.3 mmol) in concentrated H₂SO₄ at 0 °C, while keeping the reaction mixture below 5 °C. After the addition was complete, the reaction mixture was slowly warmed to room temperature overnight. Ice (50 g) was added to the mixture, the solid was collected and dried, and then washed with MTBE (50 mL) and ethyl acetate (50 mL) to give the desired product, which was a pale yellow solid (5.1 g, 50%). MS: C₈H₆FN₂O₃[M+H] + The calculated value of (ES)m / z is 197.2, and the measured value is also 197.2.

[0254] Step d: A solution of 6-fluoro-7-nitroisoindoline-1-one (11.3 g, 57 mmol) and 10% Pd / C (50% humidity, 6.2 g, 2.9 mmol, 0.05 equivalents) in THF (300 mL) was stirred overnight under a hydrogen atmosphere (balloon). The solid was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a colorless solid. This solid was purified by silica gel chromatography (100% ethyl acetate) to give the desired product as a white solid (6.4 g, 67%). MS: C8H9FN2O[M+H] + The calculated value of (ES)m / z is 168.1, and the measured value is 168.1.

[0255] Step e: A mixture of 7-amino-6-difluoro-isoindolin-1-one (4.4 g, 26 mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (7.4 g, 52 mmol) in anhydrous methanol (30 mL) was stirred overnight at 60 °C, followed by stirring at 80 °C for 5 hours. The reaction mixture was evaporated, and the residue was stirred in ethyl acetate (200 mL) at 50 °C for 30 minutes, then cooled to room temperature. The mixture was filtered and dried to give a pale yellow solid (5.0 g, 70%). MS: C 13 H 10FN₂O₄[M+H] + The calculated value of (ES)m / z is 277.2, and the measured value is also 277.2.

[0256] Step f: Anhydrous ethanol (10 mL) was added to a mixture of 3-[(7-fluoro-3-oxo-isoindoline-4-yl)amino]-4-methoxy-cyclobut-3-ene-1,2-dione (1.5 g, 5.4 mmol) and (1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propane-1-amine (1.1 g, 6.5 mmol), and the mixture was stirred overnight at 60 °C. The reactants were cooled to room temperature, dissolved in a minimal amount of dichloromethane, and adsorbed onto silica gel. The product was purified by silica gel chromatography (40% ethyl acetate in dichloromethane) to give a white solid (800 mg, 45%). 1 H NMR (400MHz, DMSO-d6) δ9.59 (s, 1H), 8.65 (s, 1H), 8.35 (d, J = 10.4Hz, 1H), 7.41 (dd, J = 11.6, 8.4Hz, 1H), 6.18 (dd, J = 4.0, 8.4Hz, 1H), 6.12 (d, J = 3.2Hz, 1H), 5.98 (d, J = 2.0Hz, 1H), 4.97 (d, J = 4.10Hz, 1H), 4.26 (s, 2H), 2.22 (s, 3H), 0.90 (s, 9H). MS:C 22 H 22 The calculated (ES) m / z value of FN3O4[MH]- is 410.0, and the measured value is also 410.0.

[0257] Example 4: Synthesis of 3-[(5,7-difluoro-3-oxo-isoindoline-4-yl)amino]-4-[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]cyclobut-3-ene-1,2-dione

[0258]

[0259] Step a: 3,5-Difluoro-2-methylbenzoic acid (5.2 g, 30.2 mmol) was dissolved in anhydrous DMF (30 mL). Anhydrous Na₂CO₃ (3.5 g, 33.2 mmol, 1.1 equivalents) was added and the reaction was stirred at room temperature for 30 minutes. Iodomethane (2.1 mL, 33.2 mmol, 1.1 equivalents) was added and the mixture was stirred at room temperature for 4 hours. The reaction mixture was then diluted with water (200 mL) and the product was extracted with Et₂O (3 × 50 mL). The combined organic layers were washed with brine (4 × 30 mL), dried over MgSO₄, filtered, and evaporated to give a yellow oil (5.4 g, 96%).

[0260] Step b: The product from step a (5.4 g, 29.0 mmol) was dissolved in carbon tetrachloride (60 mL), and N-bromosuccinimide (7.7 g, 43.5 mmol, 1.5 equivalents) was added, followed by benzoyl peroxide (1.4 g, 5.8 mmol, 0.20 equivalents). The reaction mixture was refluxed and stirred overnight, then cooled to room temperature and filtered. The filtrate was evaporated, and the residue was purified by column chromatography (silica gel, 100% hexane to 9:1 hexane:ethyl acetate) to give a yellow oil (7.4 g, 96%).

[0261] Step c: Cool NH3 (7M, 45 mL, 6.4 mmol) in methanol to 0 °C, and add the product from step b (6 g, 22.6 mmol). Stir the reaction mixture at 0 °C for 10 minutes, then leave it at room temperature overnight. Evaporate excess solvent, and dilute the residue with water (50 mL). Filter the resulting solid, wash with water (2 × 20 mL), and then with hexane (20 mL) to give the product (3.4 g, 89%). MS: C8H6F2NO[M+H] + The calculated value of (ES)m / z is 170.0, and the measured value is 170.3.

[0262] Step d: Dissolve 3.4 g (20.1 mmol) of 4,6-difluoroisoindoline-1-one from step c in concentrated H₂SO₄ (40 mL) and cool to 0 °C. Add dropwise 70% HNO₃ (1.5 mL, 24.1 mmol, 1.2 equivalences) and stir the reaction mixture at 0 °C for 10 min, then warm to room temperature for 1 hour and stir overnight. Add ice and dilute the mixture with cold water (100 mL). Filter the resulting yellow solid, wash with water (2 × 50 mL), wash with hexane (50 mL), and dry under vacuum (3.4 g, 79%). MS: C₈H₅F₂N₂O₃[M+H] + The calculated value of (ES)m / z is 215.0, and the measured value is 215.2.

[0263] Step e: 3.4 g (15.9 mmol) of 4,6-difluoro-7-nitro-isoindoline-1-one obtained from step d was added to 10% Pd / C (1.7 g, 0.8 mmol, 5% mmol) at 50% humidity under a nitrogen atmosphere with THF (50 mL). The reaction mixture was vigorously stirred for 1 day at room temperature under a H2 (balloon) atmosphere, then filtered through diatomaceous earth and evaporated to give a solid product (2.7 g, 92%). MS: C8H7F2N2O[M+H] + The calculated value of (ES)m / z is 185.1, and the measured value is 185.3.

[0264] Step f: The mixture of 7-amino-4,6-difluoro-isoindoline-1-one (2.3 g, 12.5 mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (3.5 g, 25.0 mmol, 2.0 equivalent) obtained from step e in anhydrous MeOH (15 mL) was stirred overnight at 60 °C. The reaction mixture was evaporated and the residue was diluted with MTBE:EtOAc (1:1, 200 mL) and stirred at 50 °C for 30 min, then cooled to room temperature. The solid product was filtered, washed with MTBE, dissolved in MeOH:DCM (1:1, 200 mL), and filtered through diatomaceous earth. The filtrate was evaporated to give a gray solid (2.0 g, 54%). MS: C 13 H9F2N2O4[M+H] + The calculated value of (ES)m / z is 295.1, and the measured value is 295.2.

[0265] Step g: Anhydrous methanol (30 mL) was added to a mixture of 3-[(5,7-difluoro-3-oxo-isoindoline-4-yl)amino]-4-methoxy-cyclobut-3-ene-1,2-dione (1.5 g, 5.1 mmol) and (1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propane-1-amine (852 mg, 5.1 mmol) obtained from step f, and the mixture was stirred at 60 °C for 1 day. The reactants were cooled to room temperature, dissolved in a minimal amount of dichloromethane, and adsorbed onto silica gel. The product was purified by silica gel chromatography (100:0 to 50:50 dichloromethane:ethyl acetate) to give a brown solid (1.4 g, 64%). 1 H NMR (400MHz, DMSO-d6) δ9.59(s,1H),8.92(s,1H),8.37(d,J=10.2Hz,1H),7.62(dd,J=10.9,8.6Hz,1H),6. 18(d,J=3.1Hz,1H),6.04(d,J=3.1Hz,1H),5.01(d,J=10.2Hz,1H),4.41(s,2H),2.27(s,3H),0.96(s,9H). MS:C 22 H 21 F2N3O4[MH] - The calculated value of (ES)m / z is 428.1, and the measured value is also 428.1.

[0266] Example 5: 2-[4-chloro-7-[2-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]-4-methoxy-benzoic acid

[0267]

[0268] Step a: A 1 L round-bottom flask containing 50 mL of concentrated H₂SO₄ (25.0 g, 0.149 mole) of 4-chloroisoindolin-1-one was cooled in an ice bath. A mixture of concentrated H₂SO₄ (50 mL) and 70% HNO₃ (10 mL, 0.16 mole, 0.16 equivalent) was added dropwise, and the reaction mixture was stirred at 0 °C for 2 h. The mixture was then carefully quenched with ice and diluted to 1 L with cold water. The solid was filtered, washed with water, and dried under high vacuum to give 23 g, 73% of 4-chloro-7-nitro-isoindolin-1-one. MS: C₈H₅ClN₂O₃[MH] - The calculated value of (ES)m / z is 212.0, and the measured value is also 212.0.

[0269] Step b: At room temperature, iron powder (18.2 g, 324 mmol) was added to a stirred mixture of 4-chloro-7-nitro-isoindolin-1-one (23 g, 108 mmol) and ethanol, followed by dioxane (162 mL, 648 mmol) in 4 M HCl. The reaction mixture was stirred at room temperature for 1 hour, then concentrated under vacuum. The residue was diluted with ethyl acetate, neutralized with saturated sodium bicarbonate solution, and extracted with ethyl acetate (2 × 500 mL). The combined organic layers were dried (Na₂SO₄), filtered, and concentrated under vacuum to give 7-amino-4-chloro-isoindolin-1-one (16.5 g, 72%). MS: C₈H₇ClN₂O[M+H] + The calculated value of (ES)m / z is 183.2, and the measured value is also 183.2.

[0270] Step c: To a reaction flask containing 10 mL of dioxane containing 250 mg (1.37 mmol) of 7-amino-4-chloro-isoindoline-1-one, methyl 2-bromo-5-methoxybenzoate (502 mg, 2.05 mmol), cesium carbonate (893 mg, 2.74 mmol), cuprous iodide (104 mg, 0.55 mmol), and (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (156 mg, 1.1 mmol) were added. The mixture was purged with nitrogen and then heated to 110 °C. The reaction was stirred at 110 °C for 1 hour and monitored by LC-MS. After completion, the reaction was cooled, filtered through diatomaceous earth, and washed with ethyl acetate. The crude product was purified by silica gel chromatography (0-50% ethyl acetate / hexane) to give methyl 2-(7-amino-4-chloro-1-oxo-isoindoline-2-yl)-5-methoxy-benzoate, which was a white solid (284 mg, 60%). MS: C 17 H 15ClN2O4[M+H] + The calculated value of (ES)m / zC is 347.1, and the measured value is also 347.1.

[0271] Step d: A mixture of methyl 2-(7-amino-4-chloro-1-oxo-isoindoline-2-yl)-5-methoxy-benzoate (160 mg, 0.46 mmol) and 3,4-dimethoxycyclobutane-1,2-dione (131 mg, 0.92 mmol) in anhydrous methanol (5 mL) was stirred overnight at 60 °C. The reaction mixture was evaporated, and the residue was stirred in ethyl acetate (5 mL) at 50 °C for 30 minutes, then cooled to room temperature. The mixture was filtered and dried to give the product methyl 2-[4-chloro-7-[(2-methoxy-3,4-dioxo-cyclobutyl)amino]-1-oxo-isoindoline-2-yl]-5-methoxy-benzoate, a pale yellow solid (170 mg, 81%). MS: C 22 H 17 ClN2O7[M+H] + The calculated value of (ES)m / z is 457.1, and the measured value is also 457.1.

[0272] Step e: Add 10 mL of anhydrous methanol to a mixture of methyl 2-[4-chloro-7-[(2-methoxy-3,4-dioxo-cyclobutyl)amino]-1-oxo-isoindoline-2-yl]-5-methoxy-benzoate (170 mg, 0.37 mmol) and (1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propane-1-amine (62 mg, 0.37 mmol), and stir the mixture overnight at 60 °C. The reactants were then concentrated, and the crude methyl 2-[4-chloro-7-[[2-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]-4-methoxybenzoate (218 mg, 0.37 mmol) was used for the next step without further purification.

[0273] Step f: Lithium hydroxide (78 mg, 1.85 mmol) was added to a solution of methyl 2-[4-chloro-7-[[2-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]-4-methoxybenzoate (218 mg, 0.37 mmol) in tetrahydrofuran (4.0 mL), methanol (0.5 mL), and water (0.5 mL). The resulting mixture was stirred at 60 °C for 6 hours. The reaction was acidified with 5% hydrochloric acid solution and extracted with ethyl acetate. The organic layer was dried (Na₂SO₄), filtered, and concentrated under vacuum. The crude product was purified by reversed-phase chromatography to obtain a yellow solid of 2-[4-chloro-7-[[2-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]-4-methoxybenzoic acid (37 mg, 17%). 1 H NMR (400MHz, DMSO-d6) δ9.94 (s, 1H), 9.12 (d, J = 10Hz, 1H), 7.90 (d, J = 9.2, 1H),7.66(d,J=8.8,1H),7.49(d,J=9.2,1H),7.19(d,J=2.4,1H),7.04(dd, J=8.8,2.4,1H),6.16(d,J=3.2Hz,1H),6.02(d,J=1.6Hz,1H),5.09(d,J=10 Hz, 1H), 4.78 (dd, J = 23, 5.6Hz, 2H), 3.83 (s, 3H), 2.24 (s, 3H), 0.87 (s, 9H). MS:C 30 H 28 The calculated (ES) m / z value of ClN3O7[MH]- is 576.0, and the measured value is also 576.0.

[0274] Example 6: 2-[4-chloro-7-[[2-[[(1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]-4-methoxy-benzoic acid

[0275]

[0276] Step a: Add 30.0 g (312.5 mmol, 1.3 equivalents) of 2,3-dimethylfuran and 300 mL of anhydrous Et₂O to a 1 L three-necked flask equipped with a feeding funnel, and then cool in an ice bath. Over approximately 35 minutes, add dropwise n-BuLi hexane (2.5 M, 125 mL, 312.5 mmol, 1.3 equivalents). Stir the mixture at 0 °C for 30 minutes, then at room temperature for 40 minutes, and then cool again to 0 °C. Add solid MgBr₂x Et₂O (80.6 g, 312.5 mmol, 1.3 equivalents) and stir the mixture at 0 °C for 30 minutes, then at room temperature for 20 minutes.

[0277] In a 5 L three-necked flask equipped with a mechanical stirrer and an internal thermometer, imine (45.4 g, 240.4 mmol) was dissolved in anhydrous toluene (1.2 L) and cooled to an internal temperature of -70 °C. The lithium salt solution from the previous section was added over 56 minutes, maintaining the internal temperature between -70 and -67.8 °C. After the addition was complete, the reaction mixture was stirred at -70 °C for 1 hour, then overnight at room temperature. The reaction mixture was slowly quenched with a saturated aqueous solution of NH₄Cl (400 mL) and water (400 mL), then stirred at room temperature for 15 minutes. The organic layer was then separated and washed with brine (200 mL). The combined aqueous layers were extracted with ethyl acetate (300 mL). The organic matter was dried over MgSO₄, filtered, and concentrated to give a yellow oil. The crude product was purified by silica gel chromatography (0–10% MTBE / DCM) to give the product as a single diastereomer (18.0 g, 26%). MS: C 15 H 28 NO2S[M+H] + The calculated value of (ES)m / z is 286.1, and the measured value is also 286.1.

[0278] Step b: N-[(1R)-2,2-dimethyl-1-(4,5-dimethyl-2-furanyl)propyl]-2-methyl-propane-2-sulfinamide (18 g, 63.1 mmol) from the previous step was dissolved in methanol (200 mL) and cooled in an ice bath, then a solution of 2 M HCl in diethyl ether (31.5 mL, 126.2 mmol, 2 equivalents) was added. The cooling bath was removed and the reaction mixture was stirred at room temperature for 2 hours. The solvent was removed under vacuum, and anhydrous diethyl ether (100 mL) was added to the residue. The resulting mixture was filtered. Water (100 mL) and 1 M NaOH aqueous solution (100 mL) were added to the solid. The product was extracted with dichloromethane (3 × 100 mL), and the combined organic layers were dried over MgSO4, filtered, and evaporated to give (1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propane-1-amine, which was a yellow oil (9.7 g, 85%). MS: C 11 H 17 O[(M–NH3)+H] + The calculated value of (ES)m / z is 165.1, and the measured value is also 165.1.

[0279] Step c: Anhydrous methanol (1 mL) was added to a mixture of methyl 2-[4-chloro-7-[(2-methoxy-3,4-dioxo-cyclobutyl)amino]-1-oxo-isoindoline-2-yl]-5-methoxybenzoate (60 mg, 0.13 mmol) and (1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propane-1-amine (24 mg, 0.13 mmol). The mixture was stirred at 60 °C for 3 hours. The reaction was concentrated to dryness, and the crude methyl 2-[4-chloro-7-[[2-[(1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]-4-methoxy-benzoate (78 mg, 0.13 mmol) was used in the next step without further purification.

[0280] Step d: Lithium hydroxide (27 mg, 0.65 mmol) was added to a solution of methyl 2-[4-chloro-7-[[2-[[(1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]-4-methoxybenzoate (78 mg, 0.13 mmol) in tetrahydrofuran (1.0 mL), methanol (0.1 mL), and water (0.1 mL). The resulting mixture was stirred overnight at room temperature. The reaction was acidified with 5% hydrochloric acid solution and extracted with ethyl acetate. The organic layer was dried (Na₂SO₄), filtered, and concentrated under vacuum. The crude product was purified by reversed-phase chromatography to obtain a yellow solid of 2-[4-chloro-7-[[2-[[(1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]-4-methoxybenzoic acid (12 mg, 15%). 1 HNMR (400MHz, DMSO-d6) δ9.93 (s, 1H), 9.09 (d, J = 10Hz, 1H), 7.90 (d, J = 9.2 ,1H),7.66(d,J=8.8,1H),7.49(d,J=9.2,1H),7.19(d,J=2.4,1H),7.04(d d,J=8.8,2.4,1H),6.06(d,J=3.2Hz,1H),5.04(d,J=10Hz,1H),4.83(dd,J =23,5.6Hz,2H),3.83(s,3H),2.15(s,3H),1.85(s,3H),0.87(s,9H).MS:C 31 H 30 ClN3O7[MH] - The calculated value of (ES)m / z is 590.2, and the measured value is also 590.2.

[0281] Example 7: 2-[4-chloro-7-[[2-[[(1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]-4-methyl-benzoic acid

[0282]

[0283] Step a: To a reaction vial containing 10 mL of dioxane containing 7-amino-4-chloro-isoindolin-1-one (305 mg, 1.67 mmol), methyl 2-bromo-5-methylbenzoate (575 mg, 2.51 mmol), cesium carbonate (1.63 g, 5 mmol), cuprous iodide (190 mg, 1.0 mmol), and (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (285 mg, 2.0 mmol) were added. The mixture was purged with nitrogen and then heated to 110 °C. The reaction was stirred at 110 °C for 1 hour and monitored by LC-MS. After completion, the reaction was cooled, filtered through diatomaceous earth, and washed with EtOAc. The crude product was purified by silica gel chromatography (0-50% ethyl acetate / hexane) to give methyl 2-(7-amino-4-chloro-1-oxo-isoindoline-2-yl)-5-methylbenzoate, which was a white solid (345 mg, 62%). MS: C 17 H 15 ClN2O3[M+H] + The calculated value of (ES)m / z C is 331.1, and the measured value is also 331.1.

[0284] Step b: Lithium hydroxide (874 mg, 20.83 mmol) was added to a solution of methyl 2-(7-amino-4-chloro-1-oxo-isoindoline-2-yl)-5-methylbenzoate (689 mg, 2.08 mmol) in tetrahydrofuran (10 mL), methanol (1 mL), and water (1 mL). The resulting mixture was stirred overnight at 60 °C. The reaction was then cooled, acidified to pH 5 with 1 N hydrochloric acid solution, and extracted with ethyl acetate / MeOH (10:1). The organic layer was dried (Na₂SO₄), filtered, and concentrated under vacuum. Hexane was added to the crude product, and the resulting solid was filtered and washed with hexane to give a yellow solid of 2-(7-amino-4-chloro-1-oxo-isoindoline-2-yl)-4-methylbenzoic acid (572 mg, 87%).

[0285] Step c: A solution of 2-(7-amino-4-chloro-1-oxo-isoindoline-2-yl)-4-methylbenzoic acid (570 mg, 1.80 mmol) and 3,4-dimethoxycyclobutane-1,2-dione (307 mg, 2.16 mmol) in anhydrous methanol (5 mL) was stirred overnight at 60 °C. The reaction mixture was then cooled to room temperature and filtered. The solid was then washed with EtOAc and dried to give a yellow solid of 2-[4-chloro-7-[(2-methoxy-3,4-dioxo-cyclobuten-1-yl)amino]-1-oxo-isoindoline-2-yl]-4-methylbenzoic acid (565 mg, 71%). MS: C 21 H 15 ClN2O6[M+H]+ The calculated value of (ES)m / z is 427.1, and the measured value is also 427.1.

[0286] Step d: Add anhydrous methanol (2 mL) to a mixture of 2-[4-chloro-7-[(2-methoxy-3,4-dioxo-cyclobuten-1-yl)amino]-1-oxo-isoindoline-2-yl]-4-methylbenzoic acid (60 mg, 0.14 mmol) and (1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propane-1-amine (27 mg, 0.15 mmol), and stir the mixture overnight at 60 °C. The reactants were then concentrated and the crude product was purified by reversed-phase chromatography to obtain 2-[4-chloro-7-[[2-[[(1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]-4-methylbenzoic acid (30 mg, 37%). 1 H NMR (400MHz, DMSO-d6) δ9.94 (s, 1H), 9.09 (d, J = 10Hz, 1H), 7.81 (d, J = 9.2 ,1H),7.66(d,J=8.8,1H),7.49(d,J=9.2,1H),7.42(d,J=2.4,1H),7.30(d d,J=8.8,2.4,1H),6.07(d,J=3.2Hz,1H),5.04(d,J=10Hz,1H),4.83(dd,J =23,5.6Hz,2H),2.38(s,3H),2.15(s,3H),1.85(s,3H),0.87(s,9H).MS:C 31 H 30 The calculated (ES) m / z value of ClN3O6[MH]- is 574.0, and the measured value is also 574.0.

[0287] Example 8: Synthesis of 3-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)butyl]amino]-4-[(5-fluoro-3-oxo-isoindoline-4-yl)amino]cyclobut-3-ene-1,2-dione

[0288]

[0289] Step a: Dissolve 2,2-dimethylbutyraldehyde (5.0 g, 50 mmol) and (R)-tert-butylsulfinamide (6.36 g, 52.5 mmol) in CH2Cl2 (100 mL) and add Ti(OEt)4 (85-95%, 22.81 g). ～90 mmol). The reaction mixture was stirred overnight at room temperature. The reactants were then diluted with CH2Cl2 (200 mL), and diatomaceous earth and H2O (90 mL) were added with vigorous stirring. The mixture was stirred for 5 hours, then filtered through diatomaceous earth, and the filter cake was washed with CH2Cl2. The filtrate was concentrated and purified with silica gel (1% to 30% hexane of EtOAc) to give the product.

[0290] Step b: 2-Methylfuran (5.06 mL, 56.2 mmol) in Et₂O was cooled on ice. N-BuLi (2.5 M, 22.5 mL, 56.3 mmol) was added dropwise, and the reaction mixture was stirred on ice for 15 minutes. The bath was then removed, and stirring continued at room temperature for 1 hour. The reaction mixture was then cooled on ice again, and MgBr₂ (14.5 g, 56.2 mmol) was added dropwise. The reaction mixture was stirred on ice for 20 minutes, then the bath was removed, and stirring continued at room temperature for 50 minutes. The reaction mixture was then cooled in a -78 °C bath, and (R,E)-N-(2,2-dimethylbutenyl)-2-methyl-propane-2-sulfinamide (7.6 g, 37.4 mmol) in Et₂O was added dropwise. The reaction mixture was slowly warmed to room temperature overnight. The mixture was quenched with a saturated aqueous NH₄Cl solution, stirred vigorously, diluted with H₂O, and extracted with EtOAc (3 × 150 mL). The combined organic layers were dried with MgSO4, filtered, and concentrated to obtain a crude product. This crude product was then purified on silica gel (5% to 40% hexane containing EtOAc) to obtain the pure isomer.

[0291] Step c: Dioxane (4M, 21 mL, 84 mmol) of MeOH and HCl was added to N-[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)butyl]-2-methyl-propane-2-sulfinamide (6.0 g, 21 mmol). The mixture was stirred at room temperature for 45 minutes. The reaction mixture was then concentrated and dried under vacuum to give the product.

[0292] Step d: Et3N (0.072 mL, 0.52 mmol) was added to a mixture of cyclobutene (72 mg, 0.26 mmol) and (1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)butane-1-amine hydrochloride (57 mg, 0.26 mmol) in MeOH (1.3 mL). The reaction was stirred at 60 °C for 4 hours, then overnight at room temperature. Silica gel was added to the reaction mixture, the mixture was concentrated, and the product was purified by silica gel chromatography (1% to 10% MeOH in CH2Cl2) to obtain the product. 1H NMR (400MHz, DMSO-d6) δ9.65 (s, 1H), 8.71 (s, 1H), 8.39 (d, J = 10.2Hz, 1H), 7.47 ( dd,J=11.2,8.2Hz,1H),7.33(dd,J=8.3,3.8Hz,1H),6.17(d,J=3.1Hz,1H),6.06–

[0293] 6.02(m,1H),5.11(d,J=10.2Hz,1H),4.32(s,2H),2.27(s,3H),1.36-1.21(m,2H),0.94(s,3H),0.88(s,3H),0.83(t,J=7.5Hz,

[0294] 3H). MS: C 23 H 25 FN3O4[M+H] + The calculated value of (ES)m / z C is 426.2, and the measured value is also 426.2.

[0295] Example 9: Synthesis of 3-[[(1R)-1-(5-chloro-2-furanyl)-2,2-dimethyl-propyl]amino]-4-[(7-chloro-3-oxo-isoindoline-4-yl)amino]cyclobut-3-ene-1,2-dione

[0296]

[0297] Step a: 5-Chlorofuran-2-carboxaldehyde (5.0 g, 38 mmol) and (R)-tert-butylsulfinamide (4.2 g, 35 mmol) were dissolved in CH₂Cl₂ (75 mL), and Ti(OEt)₄ (85-95%, 17.6 g, 77 mmol) was added. The reaction mixture was stirred overnight at room temperature. The reactants were then diluted with CH₂Cl₂ (150 mL), and Na₂SO₄·10H₂O (100 g) was added. The mixture was stirred for 90 minutes. The mixture was then filtered through diatomaceous earth, and the filter cake was washed with CH₂Cl₂ (200 mL). The filtrate was concentrated to give the product.

[0298] Step b: (R,E)-N-[(5-chloro-2-furanyl)methylene]-2-methyl-alkane-2-sulfinamide (7.65 g, 32.7 mmol) was dissolved in CH2Cl2 (131 mL) and cooled in a -78 °C bath under nitrogen atmosphere. t-BuMgCl2 (2 M Et2O, 33 mL, 66 mmol) was added via a feeding funnel over 30 minutes, and the reaction mixture was stirred for 4 hours. A saturated aqueous solution of NH4Cl was added, and the mixture was warmed to room temperature. H2O (50 mL) was then added, and the mixture was extracted with CH2Cl2 (2x), dried over Na2SO4, filtered, and concentrated to give a mixture of diastereomers. The crude product was adsorbed onto silica gel and purified by column chromatography (CH2Cl2 with 10% methyl tert-butyl ether). The early eluted diastereomers were collected and concentrated to give the product.

[0299] Step c: N-[(1R)-1-(5-chloro-2-furanyl)-2,2-dimethyl-propyl]-2-methyl-propane-2-sulfinamide (0.98 g, 3.4 mmol) was dissolved in MeOH (3.4 mL), and HCl (2 M HCl in Et₂O, 3.4 mL, 6.8 mmol) was added. The reaction was stirred overnight and then concentrated. Et₂O (25 mL) was added and the mixture was stirred for 30 minutes and then filtered. The solid was washed with Et₂O (2x), and then KOH aqueous solution (3 M, 5 mL) was added. The product was extracted with CH₂Cl₂ (3x). The combined organic layers were washed twice with KOH aqueous solution (1.5 M), dried over Na₂SO₄, filtered, and concentrated to give the product.

[0300] Step d: 3-[(7-chloro-3-oxo-isoindoline-4-yl)amino]-4-methoxy-cyclobut-3-ene-1,2-dione (59 mg, 0.2 mmol) and (1R)-1-(5-chloro-2-furanyl)-2,2-dimethyl-propane-1-amine (38 mg, 0.2 mmol) were combined in MeOH (0.2 mL) and the mixture was stirred overnight at room temperature. The reaction was concentrated and then purified by reversed-phase chromatography (MeCN: H2O containing 0.1% TFA as eluent) to give the product. 1 H NMR(400MHz, DMSO-d6)δ9.96(s,1H),9.17(d,J=10.0Hz,1H),8.95(s,1H),7.61(d,J=8.7Hz,1H),7.45( d,J=8.7Hz,1H),6.53–6.43(m,2H),5.17(d,J=9.9Hz,1H),4.38(s,2H),3.17(s,1H),0.99(s,9H).MS:C 21 H20 Cl2N3O4[M+H] + The calculated value of (ES)m / z is 448.1, and the measured value is also 448.1.

[0301] Example 10: (R)-3-(2,2-dimethyl-1-(5-methylfuran-2-yl)propyl)amino)-4-((5-fluoro-7-methyl-3-oxoisoindoline-4-yl)amino)cyclobut-3-ene-1,2-dione

[0302]

[0303] Step a: At room temperature, N-iodosuccinimide (4.55 g, 20.2 mmol) was added fractionally to AcOH (30 mL) of 7-amino-6-fluoroisoindoline-1-one (2.4 g, 14.4 mmol) in a water bath. The resulting mixture was stirred in a water bath for 30 min, quenched with water (20 mL), and extracted with ethyl acetate (100 mL). The organic layer was washed with brine (100 mL) and dried over MgSO4. The solvent was evaporated under reduced pressure to give a brown solid, which was purified by silica gel chromatography (0-60% ethyl acetate in hexane) to give the product. MS: C8H6FIN2O[M+H] + The calculated value of (ES)m / z is 293.0, and the measured value is 293.0.

[0304] Step b: CsF (4.57 g, 30.1 mmol), 2,4,6-trimethyl-1,3,5,2,4,6-trioxaborane (1.35 g, 22.6 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium(II) dichloride (551 mg, 0.753 mmol) were added to a dioxane solution of 7-amino-6-fluoro-4-iodoindolin-1-one (2.2 g, 7.53 mmol) in 44 mL. The resulting mixture was stirred overnight at 80 °C. The reaction mixture was then partitioned between water (100 mL) and ethyl acetate (100 mL), and the organic layer was washed with brine (80 mL) and dried over MgSO4. The solvent was evaporated under reduced pressure to give a brown solid, which was purified by silica gel chromatography (0-80% ethyl acetate in hexane) to give the product. MS: C9H9FN2O[M+H] + The calculated value of (ES)m / z is 181.1, and the measured value is also 181.1.

[0305] Step c: A mixture of 7-amino-6-fluoro-4-methylisoindolin-1-one (200 mg, 1.11 mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (189.3 mg, 1.33 mmol) in anhydrous methanol (3 mL) was stirred overnight at 60 °C, followed by stirring at 80 °C for 5 hours. The reaction mixture was evaporated and purified by silica gel chromatography (0-100% ethyl acetate in hexane) to give the product. MS: C 14 H 11 FN₂O₄[M+H] + The calculated value of (ES)m / z is 291.1, and the measured value is also 291.1.

[0306] Step d: Anhydrous methanol (2 mL) was added to a mixture of 3-((5-fluoro-7-methyl-3-oxoisoindoline-4-yl)amino)-4-methoxycyclobut-3-ene-1,2-dione (95 mg, 0.327 mmol) and (R)-2,2-dimethyl-1-(5-methylfuran-2-yl)propane-1-amine (55 mg, 0.329 mmol), and the mixture was stirred overnight at 60 °C. The reaction mixture was cooled to room temperature, dissolved in a minimal amount of dichloromethane, and adsorbed onto silica gel. The product was purified by silica gel chromatography (40% ethyl acetate in dichloromethane) to obtain the product. 1 H NMR (400MHz, DMSO-d6) δ9.46 (s, 1H), 8.61 (s, 1H), 8.24 (d, J = 10.4Hz, 1H), 7.21 (d, J = 11.6Hz, 1H), 6.07(d,J=2.4Hz,1H),5.94(d,J=2.4Hz,1H),4.90(s,2H),2.16(s,3H),2.15(s,3H),0.90(s,9H). MS:C 23 H 24 FN3O4[MH] - The calculated value of (ES)m / z is 426.2, and the measured value is also 426.2.

[0307] Example 11: (R)-3-((1-(4,5-dimethylfuran-2-yl-2,2-dimethylbutyl)amino)-4-((5-fluoro-1,1,7-trimethyl-3-oxo-isoindoline-4-yl)amino)cyclobut-3-ene-1,2-dione

[0308]

[0309] Step a: At 0°C, 4-methoxybenzylamine (34.7 g, 253 mmol) was slowly added to a 500 mL round-bottom flask containing methyl 2-(bromomethyl)-5-fluorobenzoate (25 g, 101 mmol) and THF (300 mL). The mixture was warmed to room temperature overnight. The reaction mixture was poured into a 2 L separatory funnel with ethyl acetate (300 mL) and HCl (1 N aqueous solution, 200 mL). The organic layer was washed with brine (2 × 200 mL), dried over MgSO4, filtered, concentrated, and purified by silica gel chromatography (0-30% ethyl acetate in hexane) to give 6-fluoro-2-(4-methoxybenzyl)isoindolin-1-one. MS: C 16 H 14 FNO2[M+H] + The calculated value of (ES)m / z is 272.1, and the measured value is also 272.1.

[0310] Step b: At 0 °C, NaH (7.4 g, 184.5 mmol) was added to a THF (50 mL) solution of 10 g (36.9 mmol) of 6-fluoro-2-(4-methoxybenzyl)isoindolin-1-one. The resulting mixture was stirred under nitrogen for 30 min. At 0 °C, methyl iodine (31.4 g, 221.2 mmol) was added to the reaction mixture, which was then heated overnight at 70 °C, cooled to room temperature, quenched with water (40 mL), and extracted with ethyl acetate (100 mL). The organic layer was washed with brine (100 mL) and dried over MgSO4. The solvent was concentrated to give the crude product, which was purified by silica gel chromatography (0-80% ethyl acetate in hexane) to give 6-fluoro-2-(4-methoxybenzyl)-3,3-dimethylisoindolin-1-one. MS: C 18 H 18 FNO2[M+H] + The calculated value of (ES)m / z is 300.1, and the measured value is also 300.1.

[0311] Step c: A solution of 6-fluoro-2-(4-methoxybenzyl)-3,3-dimethylisoindolin-1-one (5 g, 16.7 mmol) in TFA (25 mL) and anisole (5 mL) was heated overnight at 100 °C. The reaction mixture was poured into ice (20 g), neutralized with a saturated aqueous solution of NaHCO3 (50 mL), and extracted with ethyl acetate (100 mL). The organic layer was washed with brine (100 mL) and dried over MgSO4. The solvent was evaporated under reduced pressure to give the crude product, which was purified by silica gel chromatography (0-100% ethyl acetate in hexane) to give 6-fluoro-3,3-dimethylisoindolin-1-one. MS: C 10 H 10 FNO[M+H] +The calculated value of (ES)m / z is 180.1, and the measured value is also 180.1.

[0312] Step d: Nitric acid (1.34 mL) was added dropwise to a suspension of 6-fluoro-3,3-dimethylisoindolin-1-one (3.1 g, 17.3 mmol) in concentrated H₂SO₄ (12 mL) at 0 °C, while keeping the reaction mixture below 5 °C. After the addition was complete, the reaction mixture was slowly warmed to room temperature overnight. Ice (20 g) was added to the mixture, the solid was filtered off, and then washed with MTBE (50 mL) and ethyl acetate (50 mL) to give 6-fluoro-3,3-dimethyl-7-nitroisoindolin-1-one. MS: C 10 H9FN2O3[M+H] + The calculated value of (ES)m / z is 225.1, and the measured value is also 225.1.

[0313] Step e: A solution of 6-fluoro-3,3-dimethyl-7-nitroisoindolin-1-one (2.0 g, 8.93 mmol) and 10% Pd / C (50% humidity, 0.89 g, 0.45 mmol, 0.05 equivalents) in MeOH (50 mL) was shaken for 2 hours under a hydrogen atmosphere (35 psi). The solid was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The product was purified by silica gel chromatography (100% ethyl acetate) to give 7-amino-6-fluoro-3,3-dimethylisoindolin-1-one. MS: C 10 H 11 FN2O[M+H] + The calculated value of (ES)m / z is 195.1, and the measured value is 195.1.

[0314] Step f: At room temperature, N-iodosuccinimide (244 mg, 1.08 mmol) was added fractionally to a solution of 7-amino-6-fluoro-3,3-dimethylisoindolin-1-one (150 mg, 0.77 mmol) in AcOH (2 mL). The resulting mixture was stirred in a water bath for 30 minutes, quenched with water (1 mL), and extracted with ethyl acetate (10 mL). The organic layer was washed with brine (10 mL) and dried over MgSO4. The solvent was evaporated under reduced pressure to give the crude product, which was purified by silica gel chromatography (0-60% ethyl acetate in hexane) to give 7-amino-6-fluoro-4-iodo-3,3-dimethylisoindolin-1-one. MS: C 10 H 10 FIN2O[M+H] + The calculated value of (ES)m / z is 321.0, and the measured value is also 321.0.

[0315] Step g: To a solution of 7-amino-6-fluoro-4-iodo-3,3-dimethylisoindolin-1-one (370 mg, 1.16 mmol) in dioxane (12 mL), CsF (705 mg, 4.64 mmol), 2,4,6-trimethyl-1,3,5,2,4,6-trioxaborane (435 mg, 3.47 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium(II) dichloride (95 mg, 0.116 mmol) were added. The resulting mixture was stirred overnight at 80 °C and then cooled to room temperature. The reaction mixture was partitioned between water (20 mL) and ethyl acetate (30 mL). The organic layer was washed with brine (20 mL), dried over MgSO4, filtered, and concentrated to obtain the crude product, which was purified by silica gel chromatography (0-80% ethyl acetate in hexane) to give 7-amino-6-fluoro-3,3,4-trimethylisoindololin-1-one. MS: C 11 H 13 FN2O[M+H] + The calculated value of (ES)m / z is 209.1, and the measured value is 209.1.

[0316] Step h: A mixture of 7-amino-6-fluoro-3,3,4-trimethylisoindolin-1-one (129 mg, 0.62 mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (176.3 mg, 1.24 mmol) in anhydrous methanol (2.5 mL) was stirred overnight at 60 °C, followed by stirring at 80 °C for 5 hours. The reaction mixture was concentrated, and the crude product was purified by silica gel chromatography (0-100% ethyl acetate in hexane) to give 3-((5-fluoro-1,1,7-trimethyl-3-oxoisoindolin-4-yl)amino)-4-methoxycyclobut-3-ene-1,2-dione. MS: C 16 H 15 FN₂O₄[M+H] + The calculated value of (ES)m / z is 319.1, and the measured value is also 319.1.

[0317] Step i: Anhydrous methanol (2 mL) was added to a mixture of 3-((5-fluoro-1,1,7-trimethyl-3-oxoisoindoline-4-yl)amino)-4-methoxycyclobut-3-ene-1,2-dione (22 mg, 0.07 mmol) and (R)-1-(4,5-dimethylfuran-2-yl)-2,2-dimethylbut-1-amine (15 mg, 0.077 mmol), and the mixture was stirred overnight at 60 °C. The reaction was cooled to room temperature, dissolved in a minimal amount of dichloromethane, and adsorbed onto silica gel. It was purified by silica gel chromatography (40% ethyl acetate in dichloromethane) to give the title compound. 1H NMR (400MHz, Cd3OD) δ7.21(d,J=12Hz,1H),6.04(s,1H),5.16(d,J=4.10Hz,1H),2.47(s,3H),2.19(s, 3H), 1.92 (s, 3H), 1.60 (s, 6H), 1.40 (q, J = 7.6Hz, 2H), 1.03 (s, 3H), 0.97 (s, 3H), 0.91 (t, J = 7.6Hz, 3H). MS:C 27 H 32 FN3O4[MH] - The calculated value of (ES)m / z is 482.2, and the measured value is also 482.2.

[0318] Example 12: Synthesis of (R)-3-((7-chloro-2-(3-methyl-1H-pyrazol-5-yl)-3-oxoisoindoline-4-yl)amino)-4-((2,2-dimethyl-1-(5-methylfuran-2-yl)propyl)amino)cyclobut-3-ene-1,2-dione.

[0319]

[0320] Step a: Triethylamine (0.5 mL, 3.56 mmol) was added to an anhydrous tetrahydrofuran solution (3 mL) of methyl 2-(bromomethyl)-3-chloro-6-nitrobenzene (500 mg, 1.62 mmol) and 3-methyl-1H-pyrazole-5-amine (158 mg, 1.62 mmol). The resulting reaction solution was heated to 60 °C for 1 hour in a sealed 40 mL reaction flask. A solid precipitate formed during the reaction. The reaction mixture was then cooled to room temperature and filtered. The solid was washed with dichloromethane to give the product. MS: C 12 H9ClN4O3[M+H] + The calculated value of (ES)m / z is 293.0, and the measured value is also 293.0.

[0321] Step b: At room temperature, iron powder (203 mg, 3.60 mmol) was added to a stirred mixture of 4-chloro-2-(3-methyl-1H-pyrazol-5-yl)-7-nitroisoindoline-1-one (266 mg, 0.91 mmol) and ethanol, followed by dioxane (0.91 mL, 3.64 mmol) containing 4 M HCl. The reaction mixture was stirred at room temperature for 1 hour and then concentrated to dryness. The residue was diluted with ethyl acetate and neutralized with saturated sodium bicarbonate solution, and extracted with ethyl acetate (2 × 5 mL). The combined organic layers were dried (Na₂SO₄), filtered, and concentrated under vacuum. The crude product was purified by silica gel column chromatography (0–100% ethyl acetate in hexane) to give the product. MS: C 12 H11 ClN4O[M+H] + The calculated value of (ES)m / z is 263.0, and the measured value is also 263.0.

[0322] Step c: To a methanol (1 mL) suspension of 7-amino-4-chloro-2-(3-methyl-1H-pyrazol-5-yl)isoindolin-1-one (200 mg, 0.76 mmol) and 3,4-dimethoxycyclobut-3-en-1-one (130 mg, 0.91 mmol), dioxane (0.19 mL, 0.76 mmol) in 4 M HCl was added. The reaction mixture was heated to 60 °C and stirred for 1 hour. It was then cooled to room temperature, filtered, and washed with methanol to give the product. MS: C 17 H 13 ClN4O4[M+H] + The calculated value of (ES)m / z is 373.0, and the measured value is also 373.0.

[0323] Step d: To a methanol (1 mL) suspension of 3-((7-chloro-2-(3-methyl-1H-pyrazol-5-yl)-3-oxoisoindoline-4-yl)amino)-4-methoxycyclobut-3-ene-1,2-dione (100 mg, 0.27 mmol) and (R)-2,2-dimethyl-1-(5-methylfuran-2-yl)propane-1-amine (45 mg, 0.27 mmol) and triethylamine (0.04 mL, 0.27 mmol), (R)-2,2-dimethyl-1-(5-methylfuran-2-yl)propane-1-amine (45 mg, 0.27 mmol) and (0.04 mL, 0.27 mmol), (R)-2,2-dimethyl-1-(5-methylfuran-2-yl)propane-1-amine ... 1 H NMR (400MHz, DMSO-d6) δ12.33(s,1H),10.00(s,1H),9.13(d,J=10Hz,1H),7.63(d,J=10Hz,1H),7.46(d,J=10Hz,1H),6.56(s ,1H),6.20(d,J=3.2Hz,1H),6.04(d,J=3.2Hz,1H),5.12(d,J=10Hz,1H),4.85(s,2H),2.28(s,3H),2.25(s,3H),0.97(s,9H). MS:C 26 H 26 The calculated (ES) m / z value of ClN5O4[MH]- is 506.1, and the measured value is also 506.1.

[0324] Example 13: Synthesis of (R)-3-((7-chloro-5-fluoro-1,1-dimethyl-3-oxoisoindoline-4-yl)amino)-4-((2,2-dimethyl-1-(5-methylfuran-2-yl)propyl)amino)cyclobut-3-ene-1,2-dione

[0325]

[0326] Step a: A mixture of methyl 2-bromo-5-fluorobenzoate (5.00 g, 21.5 mmol) and cuprous cyanide (2.12 g, 23.6 mmol) in DMF was heated at 90 °C for 1 day, then cooled to room temperature, diluted with ethyl acetate (300 mL), and filtered. The filtrate was washed with brine (5 × 50 mL) and then with a saturated aqueous solution of NaHCO3 (50 mL). The organic layer was dried over MgSO4, filtered, and concentrated under vacuum. The product was used in the next step without further purification. MS: C9H6FNO2[M+H] + The calculated value of (ES)m / z is 180.0, and the measured value is also 180.0.

[0327] Step b: At 0 °C, lithium hydroxide monohydrate (1.11 g, 26.5 mmol) was added to a stirred solution of methyl 2-cyano-5-fluorobenzoate (3.85 g, 21.5 mmol) in tetrahydrofuran (30 mL) and water (3 mL). The reaction was heated to room temperature and stirred for 1 hour. The solvent was then evaporated, and the residue was diluted with water (100 mL) and 2M HCl aqueous solution (20 mL). The solid was collected by filtration and dried under vacuum to obtain the desired product. MS: C8H4FNO2[M+H] + The calculated value of (ES)m / z is 166.0, and the measured value is also 166.0.

[0328] Step c: At -78 °C, a solution of diethyl ether containing 1.6 M lithium (25.74 mL, 41.2 mmol) was added dropwise to an anhydrous tetrahydrofuran (105 mL) stirred solution of 2-cyano-5-fluorobenzoic acid (1.70 g, 10.3 mmol). The mixture was stirred at -78 °C for 1 hour, then slowly warmed to room temperature, quenched with a saturated aqueous solution of ammonium chloride, and extracted with ethyl acetate. The organic layer was purified by silica gel chromatography (0–100% ethyl acetate in hexane) to give 6-fluoro-3,3-dimethylisoindoline-1-one. MS: C 10 H 10 FNO[M+H] + The calculated value of (ES)m / z is 180.0, and the measured value is also 180.0.

[0329] Step d: Cool a reaction vial containing 1 mL of concentrated H₂SO₄ (620 mg, 3.46 mmol) in an ice bath. Add dropwise a mixture of 1 mL of concentrated H₂SO₄ (0.25 mL, 3.8 mmol) of 70% HNO₃, and stir the reaction mixture at 0 °C for 2 h. Then carefully quench the mixture with ice and dilute to 10 mL with cold water. Filter the solid, wash with water, and dry under vacuum to give 6-fluoro-3,3-dimethyl-7-nitroisoindolin-1-one. MS: C 10 H9FN2O3[M+H] + The calculated value of (ES)m / z is 225.0, and the measured value is also 225.0.

[0330] Step e: Iron powder (0.58 g, 10.38 mmol) and ammonium chloride (1.90 g, 34.6 mmol) were added to a solution of 6-fluoro-3,3-dimethyl-7-nitroisoindolin-1-one (0.56 g, 2.50 mmol) in ethanol (10 mL) and water (1 mL) at room temperature. The reaction mixture was heated to 90 °C and stirred for 1 hour. The reaction was then cooled to room temperature, filtered through diatomaceous earth, and washed with methanol (20 mL). The filtrate was concentrated to dryness, and the residue was diluted with ethyl acetate and washed with water and brine. The combined organic layers were dried (Na₂SO₄), filtered, concentrated under vacuum, and purified by silica gel chromatography (0-100% ethyl acetate in hexane) to give 7-amino-6-fluoro-3,3-dimethylisoindolin-1-one. MS: C 10 H 11 FN2O[M+H] + The calculated value of (ES)m / z is 195.0, and the measured value is 195.0.

[0331] Step f: N-chlorosuccinimide (80 mg, 0.59 mmol) was added to a solution of 7-amino-6-fluoro-3,3-dimethylisoindolin-1-one (116 mg, 0.59 mmol) in acetic acid (1 mL) at room temperature. The reaction mixture was warmed to 45 °C and stirred overnight. It was then cooled to room temperature, diluted with ethyl acetate, and washed with water and brine. The combined organic layers were dried (Na₂SO₄), filtered, and concentrated under vacuum. The crude product was purified by silica gel chromatography (0–30% ethyl acetate in hexane) to give 7-amino-4-chloro-6-fluoro-3,3-dimethylisoindolin-1-one. MS: C 10 H 10 ClFN2O[M+H] + The calculated value of (ES)m / z is 229.0, and the measured value is also 229.0.

[0332] Step g: To a methanol (3 mL) suspension of 7-amino-4-chloro-6-fluoro-3,3-dimethylisoindolin-1-one (73 mg, 0.32 mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (540.38 mmol), dioxane (0.08 mL, 0.32 mmol) in 4 M HCl was added. The reaction mixture was warmed to 60 °C and stirred for 1 hour. It was then cooled to room temperature and diluted with dichloromethane (2 mL) to produce a clear solution. This solution was concentrated under vacuum. The crude product was purified by silica gel chromatography (0–10% methanol in dichloromethane) to provide the desired product. MS: C 15 H 12 ClFN2O4[M+H] + The calculated value of (ES)m / z is 338.0, and the measured value is also 338.0.

[0333] Step h: To a methanol (2 mL) suspension of 3-((7-chloro-5-fluoro-1,1-dimethyl-3-oxoisoindoline-4-yl)amino)-4-methoxycyclobut-3-en-1,2-dione (71 mg, 0.21 mmol), (R)-2,2-dimethyl-1-(5-methylfuran-2-yl)propane-1-amine (35 mg, 0.21 mmol) and triethylamine (0.03 mL, 0.21 mmol) were added. The resulting mixture was stirred overnight at room temperature and then diluted with dichloromethane. Purification was then performed by silica gel chromatography (0–10% methanol in dichloromethane) to give the title compound. 1 H NMR (400MHz, DMSO-d6) δ9.61(s,1H),8.97(s,1H),8.40(d,J=10Hz,1H),7.62(d,J=10Hz,1H),6.10( d,J=2.6Hz,1H),5.95(d,J=2.6Hz,1H),4.92(d,J=10Hz,1H),2.19(s,3H),1.46(s,6H),0.87(s,9H). MS:C 24 H 25 The calculated (ES) m / z value of ClFN3O4[MH]- is 472.0, and the measured value is also 472.0.

[0334] Example 14: Synthesis of 3-[(7-chloro-5-fluoro-1,1-dimethyl-3-oxo-isoindoline-4-yl)amino]-4-[[(1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propyl]amino]cyclobut-3-ene-1,2-dione

[0335]

[0336] To a methanol (2 mL) suspension of 3-((7-chloro-5-fluoro-1,1-dimethyl-3-oxoisoindoline-4-yl)amino)-4-methoxycyclobut-3-ene-1,2-dione (15 mg, 0.04 mmol), (R)-1-(4,5-dimethylfuran-2-yl)-2,2-dimethylpropane-1-amine (11 mg, 0.05 mmol) and triethylamine (0.01 mL, 0.05 mmol) were added. The resulting mixture was stirred overnight at room temperature and concentrated to dryness. The crude product was purified by reversed-phase chromatography to obtain the final product. 1 H NMR (400MHz, DMSO-d6) δ9.68(s,1H),9.06(s,1H),8.44(d,J=10Hz,1H),7.70(d,J=10Hz,1 H), 6.08 (s, 1H), 4.94 (d, J = 10Hz, 1H), 2.17 (s, 3H), 1.87 (s, 3H), 1.54 (s, 6H), 0.94 (s, 9H). MS:C 25 H 27 ClFN3O4[MH] - The calculated value of (ES)m / z is 486.0, and the measured value is also 486.0.

[0337] Example 15: Synthesis of 3-[[7-chloro-3-oxo-2-[2-(5-oxo-1H-tetrazol-4-yl)ethyl]isoindoline-4-yl]amino]-4-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]cyclobut-3-ene-1,2-dione

[0338]

[0339] Step a: Triethylamine (10 ml, 71.3 mmol) was added to a mixture of methyl 2-(bromomethyl)-3-chloro-6-nitrobenzoate (10.0 g, 32.4 mmol) and ethyl 3-aminopropionate hydrochloride (5.5 g, 35.6 mmol) in tetrahydrofuran (120 ml). The reaction mixture was stirred overnight at room temperature. The reaction mixture was then diluted with ethyl acetate and washed with water and brine. The combined organic layers were dried over (Na₂SO₄), filtered, and concentrated under vacuum to give the product, which was ready for use without further purification. MS: C 13 H 13 ClN2O5[M+H] + The calculated value of (ES)m / z is 313.0, and the measured value is also 313.0.

[0340] Step b: At room temperature, iron powder (6.0 g, 97.2 mmol) and ammonium chloride (9.0 g, 162 mmol) were added to a solution of ethyl 3-(4-chloro-7-nitro-1-oxoisoindoline-2-yl)propionate (10.1 g, 32.4 mmol) in ethanol (90 mL) and water (10 mL). The reaction mixture was heated to 90 °C and stirred for 1 hour. It was then cooled to room temperature, filtered through diatomaceous earth, and washed with methanol (120 mL). The filtrate was concentrated to dryness, and the residue was diluted with ethyl acetate, washed with water, and then washed with brine. The combined organic layers were dried (Na₂SO₄), filtered, and concentrated under vacuum to give the product, which was ready for use without further purification. MS: C 13 H 15 ClN2O3[M+H] + The calculated value of (ES)m / z is 283.0, and the measured value is also 283.0.

[0341] Step c: At room temperature, di-tert-butyl dicarbonate (12.67 g, 58.0 mmol) and 4-di(methylamino)pyridine (142 mg, 1.16 mmol) were added to a solution of ethyl 3-(7-amino-4-chloro-1-oxoisoindoline-2-yl)propionate (6.60 g, 23.2 mmol) in tetrahydrofuran (40 mL). The reaction mixture was warmed to 100 °C and stirred overnight. It was then cooled to room temperature, diluted with saturated aqueous NaHCO3 solution (100 mL), and stirred for 20 min. The reaction mixture was then diluted with ethyl acetate and washed successively with water and brine. The combined organic layers were dried (Na2SO4), filtered, and concentrated under vacuum. The crude product was purified by silica gel chromatography (0–40% ethyl acetate in hexane) to provide the product. MS: C 23 H 31 ClN2O7[M+Na] + The calculated value of (ES)m / z is 505.0, and the measured value is also 505.0.

[0342] Step d: Lithium hydroxide monohydrate (1.9 g, 46.2 mmol) was added to a solution of ethyl 3-[7-[bis(tert-butoxycarbonyl)amino]-4-chloro-1-oxo-isoindoline-2-yl]propionate (7.43 g, 15.4 mmol) in tetrahydrofuran (40 mL), methanol (4 mL), and water (4 mL). The reaction mixture was stirred overnight and then concentrated to dryness. The residue was acidified to pH 4 with 1 M HCl. The mixture was then extracted with ethyl acetate and washed with water and brine. The combined organic layers were dried (Na₂SO₄), filtered, and concentrated under vacuum to give the product, which was ready for use without further purification. MS: C 21 H 27 ClN2O7[M+H] +The calculated value of (ES)m / z is 455.0, and the measured value is also 455.0.

[0343] Step e: At 0 °C, oxaloyl chloride (1.3 mL, 15.4 mmol) was added dropwise to a suspension of 3-[7-[bis(tert-butoxycarbonyl)amino]-4-chloro-1-oxo-isoindoline-2-yl]propionic acid (4.66 g, 10.2 mmol) in dichloromethane (40 mL). After the addition was complete, two drops of DMF were added. The reaction mixture was stirred at 0 °C for 10 minutes, then warmed to room temperature for 3 hours. The resulting solution was concentrated to dryness. The residue was dissolved in dichloromethane (40 mL) and concentrated again to dryness to remove excess oxaloyl chloride. The crude product was used in the next step without further purification.

[0344] Step f: At room temperature, azidotrimethylsilane was added in a single step to the above acyl chloride under a nitrogen atmosphere. The mixture was heated to 100°C for 2 hours, then cooled to room temperature. The mixture was then concentrated to dryness to remove excess azidotrimethylsilane. The crude product was diluted with ethyl acetate and acidified to pH 3 with 1M HCl aqueous solution. The organic layer was washed with water, then with brine. The combined organic layers were dried (Na₂SO₄), filtered, and concentrated under vacuum. The crude product was purified by silica gel chromatography (0-100% ethyl acetate in hexane) to provide the product. MS: C 21 H 27 ClN6O6[M+Na] + The calculated value of (ES)m / z is 517.0, and the measured value is also 517.0.

[0345] Step g: At room temperature, trifluoroacetic acid (0.25 ml) was added to a solution of N-tert-butoxycarbonyl-N-[7-chloro-3-oxo-2-[2-(5-oxo-1H-tetrazol-4-yl)ethyl]isoindoline-4-yl]carbamate tert-butyl (135 mg, 0.27 mmol) in dichloromethane (1 ml). The reaction mixture was stirred at room temperature for 1 hour and then neutralized with a saturated aqueous solution of NaHCO3. The mixture was extracted with dichloromethane and the organic layer was washed with water and brine. The combined organic layers were dried (Na2SO4), filtered, and concentrated under vacuum. The crude product was purified by silica gel chromatography (0-100% ethyl acetate in hexane) to provide the product. MS: C 11 H 11 ClN6O2[M+H] + The calculated value of (ES)m / z is 295.0, and the measured value is also 295.0.

[0346] Step h: To a methanol (1 mL) suspension of 7-amino-4-chloro-2-(2-(5-oxo-4,5-dihydro-1H-tetrazol-1-yl)ethyl)isoindolin-1-one (60 mg, 0.21 mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (44 mg, 0.31 mmol), dioxane (0.05 mL, 0.21 mmol) containing 4 M HCl was added. The resulting clear solution was warmed to 60 °C and stirred for 1 hour, during which time a solid precipitated. The reaction mixture was cooled to room temperature, the solid was filtered off, and the mixture was washed with ethyl acetate (2 mL) to provide the product. MS: C 16 H 13 ClN6O5[MH] - The calculated value of (ES)m / z is 403.0, and the measured value is also 403.0.

[0347] Step i: To a methanol (2 mL) suspension of 3-((7-chloro-3-oxo-2-(2-(5-oxo-4,5-dihydro-1H-tetrazol-1-yl)ethyl)isoindoline-4-yl)amino)-4-methoxycyclobut-3-ene-1,2-dione (40 mg, 0.10 mmol) and (R)-2,2-dimethyl-1-(5-methylfuran-2-yl)propane-1-amine (20 mg, 0.12 mmol) and one drop of triethylamine were added. The resulting mixture was stirred overnight at room temperature and then concentrated to dryness. The crude product was purified by silica gel chromatography (0-100% ethyl acetate in hexane) to give the product. 1 H NMR (400MHz, DMSO-d6) δ9.84(s,1H),9.11(d,J=10Hz,1H),7.58(d,J=8Hz,1H),7.42(d,J=8Hz,1H),6.18(d,J=2.6Hz,1H),6.03(d,J =2.6Hz, 1H), 5.10 (d, J = 10.4Hz, 1H), 4.54 (s, 2H), 4.22 (t, J = 5.6, 5.6Hz, 2H), 3.85 (t, J = 5.6, 5.6Hz, 2H), 2.27 (s, 3H), 0.95 (s, 9H). MS:C 25 H 26 The calculated (ES) m / z value of ClN7O5[MH]- is 538.0, and the measured value is also 538.0.

[0348] Example 16: Synthesis of (R)-3-((7-chloro-2-(2-(4-methyl-5-oxo-4,5-dihydro-1H-tetrazol-1-yl)ethyl)-3-oxoisoindoline-4-yl)amino)-4-((2,2-dimethyl-1-(5-methylfuran-2-yl)propyl)amino)cyclobut-3-ene-1,2-dione

[0349]

[0350] Step a: At room temperature, potassium carbonate (70 mg, 0.51 mmol) and iodomethane were added to a DMF (1 mL) solution of tert-butyl N-tert-butoxycarbonyl-N-[7-chloro-3-oxo-2-[2-(-oxo-1H-tetrazol-4-yl)ethyl]isoindoline-4-yl] (100 mg, 0.20 mmol). The reaction mixture was stirred at room temperature for 2 hours, then quenched with water. The solid precipitate was collected and filtered, then washed with water and hexane. The collected solid was dried under vacuum to provide the product. MS: C 22 H 29 ClN6O6[M+H] + The calculated value of (ES)m / z is 509.0, and the measured value is also 509.0.

[0351] Step b: At room temperature, trifluoroacetic acid (0.25 ml) was added to a solution of N-tert-butoxycarbonyl-N-[7-chloro-2-[2-(4-methyl-5-oxo-tetrazol-1-yl)ethyl]-3-oxo-isoindoline-4-yl]carbamate (80 mg, 0.16 mmol) in dichloromethane (1 ml). The reaction mixture was stirred at room temperature for 1 hour and then neutralized with a saturated aqueous solution of NaHCO3. The mixture was extracted with dichloromethane, and the organic layer was washed with water and brine. The combined organic layers were dried (Na2SO4), filtered, and concentrated under vacuum. The crude product was purified by silica gel chromatography (0-100% ethyl acetate in hexane) to provide the product. MS: C 12 H 13 ClN6O2[M+H] + The calculated value of (ES)m / z is 309.0, and the measured value is also 309.0.

[0352] Step c: To a methanol (1 mL) suspension of 7-amino-4-chloro-2-(2-(4-methyl-5-oxo-4,5-dihydro-1H-tetrazol-1-yl)ethyl)isoindolin-1-one (31 mg, 0.10 mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (21 mg, 0.15 mmol), dioxane (0.025 mL, 0.10 mmol) in 4 M HCl was added. The resulting clear solution was warmed to 60 °C and stirred for 1 hour. The reaction mixture was cooled to room temperature, the solid precipitate was filtered off, and the mixture was washed with ethyl acetate (2 mL) to provide the product. MS: C 17 H 15 ClN6O5[MH] - The calculated value of (ES)m / z is 417.0, and the measured value is also 417.0.

[0353] Step d: To a methanol (2 mL) suspension of 3-((7-chloro-2-(2-(4-methyl-5-oxo-4,5-dihydro-1H-tetrazol-1-yl)ethyl)-3-oxoisoindoline-4-yl)amino)-4-methoxycyclobut-3-ene-1,2-dione (32 mg, 0.076 mmol) and one drop of triethylamine, (R)-2,2-dimethyl-1-(5-methylfuran-2-yl)propane-1-amine (15 mg, 0.09 mmol). The resulting mixture was stirred overnight at room temperature and concentrated to dryness. The crude product was purified by silica gel chromatography (0-100% ethyl acetate in hexane) to give the product. 1 H NMR (400MHz, DMSO-d6) δ9.85 (s, 1H), 9.10 (d, J = 10Hz, 1H), 7.59 (d, J = 8.8Hz, 1H), 7.44 (d, J = 8.8Hz, 1H), 6.18 (d, J = 2.8Hz, 1H), 6.03 (d ,J=2.8Hz,1H),5.10(d,J=10.4Hz,1H),4.54(s,2H),4.22(t,J=5.6,5.6Hz,2H),3.86(t,J=5.6,5.6Hz,2H),2.27(s,3H),0.95(s,9H). MS:C 26 H 28 ClN7O5[MH] - The calculated value of (ES)m / z is 552.0, and the measured value is also 552.0.

[0354] Example 17: Synthesis of 2-[4-chloro-7-[[2-[[(1R)-1-(5-chloro-2-furanyl)-2,2-dimethyl-propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]-6-methoxy-pyridine-3-carboxylic acid

[0355]

[0356] Step a: To a reaction vial containing 2.0 mL of dioxane containing 365 mg (2.0 mmol) of 7-amino-4-chloro-isoindoline-1-one, methyl 2-chloro-6-methoxynicotinate (603 mg, 3.0 mmol), cesium carbonate (1.3 g, 4.0 mmol), cuprous iodide (152 mg, 80 mmol), and (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (227 mg, 1.6 mmol) were added. The mixture was purged with nitrogen and then heated to 110 °C. The reaction was stirred at 110 °C and monitored by LC-MS. After completion, the reaction was cooled and filtered through diatomaceous earth and washed with ethyl acetate. The crude product was purified by silica gel chromatography (0–50% ethyl acetate / hexane) to give the product.

[0357] Step b: Lithium hydroxide (533 mg, 12.7 mmol) was added to a solution of methyl 2-(7-amino-4-chloro-1-oxo-isoindoline-2-yl)-6-methoxy-pyridine-3-carboxylic acid (440 mg, 1.27 mmol) in tetrahydrofuran (5.0 mL), methanol (0.5 mL), and water (0.5 mL). The resulting mixture was stirred at room temperature. After completion, the reaction was acidified to pH 5–7 with 1N aqueous HCl and extracted with ethyl acetate. The organic layer was washed with brine, dried (Na₂SO₄), filtered, and concentrated under vacuum to give the product.

[0358] Step c: A mixture of 2-(7-amino-4-chloro-1-oxo-isoindoline-2-yl)-6-methoxy-pyridine-3-carboxylic acid (334 mg, 1.00 mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (156 mg, 1.10 mmol) in anhydrous methanol (5 mL) was stirred at 60 °C for 3 hours. The reaction mixture was then filtered, and the solid was washed with ethyl acetate and dried to give the product.

[0359] Step d: Et3N (0.08 mL, 0.6 mmol) was added to a mixture of 2-[4-chloro-7-[(2-methoxy-3,4-dioxo-cyclobuten-1-yl)amino]-1-oxo-isoindoline-2-yl]-6-methoxy-pyridine-3-carboxylic acid (97 mg, 0.22 mmol) and (1R)-1-(5-chloro-2-furanyl)-2,2-dimethyl-propane-1-amine (45 mg, 0.24 mmol) in MeOH (3.0 mL). The reaction was stirred at 60 °C for 4 hours, and then concentrated. The crude product was purified by reversed-phase chromatography (MeCN: H2O containing 0.1% TFA as eluent) to obtain the product. 1H NMR (400MHz, methanol-d4) δ8.20(d,J=8.5Hz,1H),7.94(d,J=8.9Hz,1H),7.62(d,J=8.8,1H),6.81(d,J=8 .6Hz,1H),6.39(s,1H),6.25(d,J=3.5Hz,1H),5.28(s,1H),5.17(s,2H),4.02(s,3H),1.06(s,9H). MS:C 28 H 23 The calculated (ES) m / z value of Cl2N4O7[MH]- is 597.1, and the measured value is also 597.1.

[0360] Example 18: Synthesis of 2-[4-chloro-7-[[2-[[(1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propyl]amino]-3,4-dioxo-cyclobuten-1-yl]amino]-1-oxo-isoindoline-2-yl]-6-methoxy-pyridine-3-carboxylic acid

[0361]

[0362] Et3N (0.08 mL, 0.6 mmol) was added to a mixture of 2-[4-chloro-7-[(2-methoxy-3,4-dioxo-cyclobuten-1-yl)amino]-1-oxo-isoindoline-2-yl]-6-methoxy-pyridine-3-carboxylic acid (90 mg, 0.15 mmol) and (1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propane-1-amine hydrochloride (45 mg, 0.23 mmol) in MeOH (3.0 mL). The reaction was stirred at 60 °C for 4 hours, and then concentrated. The crude product was purified by reversed-phase chromatography (MeCN: H2O containing 0.1% TFA as eluent) to obtain the product. 1 H NMR (400MHz, methanol-d4) δ8.19(d,J=8.5Hz,1H),7.92(d,J=9.1Hz,1H),7.61(d,J=8.8Hz,1H),6.80(d, J=8.6Hz,1H),6.06(s,1H),5.24–5.09(m,3H),4.02(s,3H),2.18(s,3H),1.91(s,3H),1.04(s,9H). MS:C 30 H 28 The calculated (ES) m / z value of ClN4O7[MH]- is 591.2, and the measured value is 591.1.

[0363] Example 19: Synthesis of 3-[[(1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propyl]amino]-4-[(5-fluoro-3-oxo-isoindoline-4-yl)amino]cyclobut-3-ene-1,2-dione

[0364]

[0365] Et3N (0.08 mL, 0.6 mmol) was added to a mixture of 3-[(5-fluoro-3-oxo-isoindoline-4-yl)amino]-4-methoxy-cyclobut-3-ene-1,2-dione (81 mg, 0.30 mmol) and (1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)butane-1-amine hydrochloride (72 mg, 0.33 mmol) in MeOH. The reaction was stirred at 60 °C for 4 hours. The reaction was concentrated and then purified by reversed-phase chromatography (MeCN: H2O containing 0.1% TFA as eluent) to give the product. 1 H NMR (400MHz, DMSO-d6) δ9.64(s,1H),8.72(s,1H),8.39(d,J=10.2Hz,1H),7.47(dd,J=11.2,8.2Hz,1H),7.33(d d,J=8.3,3.8Hz,1H),6.07(s,1H),4.97(d,J=10.2Hz,1H),4.32(s,2H),2.18(s,3H),1.87(s,3H),0.95(s,9H).

[0366] Example 20: Synthesis of 3-[(7'-chloro-5'-fluoro-3'-oxo-spiro[cyclopentane-1,1'-isoindoline]-4'-yl)amino]-4-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]cyclobut-3-en-1,2-dione

[0367]

[0368] Step a: A mixture of methyl 2-bromo-5-fluorobenzoate (2.5 g, 10.8 mmol), 2-cyclopentenyl-4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane (2.3 g, 11.9 mmol), and K₂CO₃ (3.7 g, 27.0 mmol) in 1,2-dimethoxyethane (27 mL) and H₂O (3.0 mL) was purged with N₂ for 2 min. Then, Pd(PPh₃)₄ (0.62 g, 0.54 mmol) was added at room temperature. The resulting mixture was heated to 95 °C for 14 h. After the reaction was complete, the mixture was cooled to room temperature. The reaction mixture was diluted with EtOAc (100 mL), the organic layer was washed with H₂O, then with brine, dried over Na₂SO₄, filtered, and concentrated. The crude compound was purified by silica gel chromatography (0-20% ethyl acetate in hexane) to obtain methyl 2-cyclopentenyl-5-fluorobenzoate.

[0369] Step b: Add PtO2 (448 mg, 2.0 mmol) to a solution of methyl 2-cyclopentenyl-5-fluorobenzoate (2.2 g, 10.0 mmol) in MeOH (25 mL). Shake at H2 (40 psi) for 3 hours. Filter the mixture through diatomaceous earth and wash with MeOH (40 mL), then concentrate the filtrate under reduced pressure. The crude product can be used directly for the next step without further purification.

[0370] Step c: At room temperature, LiOH·2H2O (1.89 g, 61.0 mmol) was added to a stirred solution of methyl 2-cyclopentyl-5-fluorobenzoate (2.0 g, 14.9 mmol) in THF / H2O (20:6 mL). The reaction mixture was stirred for 16 hours. After completion, the reaction was quenched with 2N HCl aqueous solution (4 mL) to adjust the pH to 7. The aqueous solution was extracted with ethyl acetate (2 × 75 mL), and the combined organic layers were washed with brine and subjected to Na2SO4. 4, Dry, filter, and concentrate. The crude product can be used directly for the next step without further purification. MS: C 12 H 13 FO2[M+H] + The calculated value of (ES)m / z is 209.1, and the measured value is also 209.1.

[0371] Step d: DMF (2 drops) was added to a stirred solution of 2-cyclopentyl-5-fluorobenzoic acid (3.0 g, 14.4 mmol) in dichloromethane (30 mL). Then, oxaloyl chloride (2.27 g, 18.0 mmol) was added dropwise over 5 minutes, and the reaction mixture was stirred at room temperature for 16 hours. After the reaction was complete, the solvent was removed under reduced pressure, and the residue was dried under vacuum for 2 hours to obtain 2-cyclopentyl-5-fluorobenzoyl chloride.

[0372] At 0 °C, 5 mL of ethyl acetate containing 2-cyclopentyl-5-fluorobenzoyl chloride (3.0 g, 13.2 mmol) obtained above was added to a cold solution of O-methylhydroxylamine hydrochloride (1.32 g, 15.8 mmol) and K₂CO₃ (3.6 g, 26.4 mmol) in EtOAc and H₂O (32:10 mL). The reaction mixture was then stirred at room temperature for 14 hours. After the reaction was complete, the mixture was extracted with ethyl acetate (2 × 50 mL), the combined organic layers were washed with aqueous brine, dried over Na₂SO₄, filtered, and concentrated under vacuum. The crude compound was purified by silica gel chromatography (10–40% ethyl acetate in hexane) to give 2-cyclopentyl-5-fluoro-N-methoxybenzamide. MS: C 13 H 16 FNO2[M+H] + The calculated value of (ES)m / z C is 238.2, and the measured value is 238.1.

[0373] Step e: At room temperature, 2-iodobiphenyl (468 mg, 1.68 mmol) was added to a mixture of 2-cyclopentyl-5-fluoro-N-methoxybenzamide (2.0 g, 8.4 mmol) and mCPBA (2.16 g, 12.6 mmol) in hexafluoropropanol (3.5 mL). The reaction mixture was stirred at room temperature for 1 hour. After completion, the reaction mixture was quenched with a saturated aqueous solution of NaHCO3 and diluted with ethyl acetate (100 mL). The organic layer was washed with H2O, then with a brine solution, dried over Na2SO4, filtered, and concentrated. The crude compound was purified by silica gel chromatography (10–60% ethyl acetate in hexane) to give 5'-fluoro-2'-methoxyspiro[cyclopentane-1,1'-isoindoline]-3'-one. MS: C 13 H 14 FNO2[M+H] + The calculated value of (ES)m / z C is 236.1, and the measured value is 236.0.

[0374] Step f: At room temperature, 60% NaH (185 mg, 8.08 mmol) was added to a stirred solution of 5'-fluoro-2'-methoxyspiro-[cyclopentane-1,1'-isoindoline]-3'-one (0.95 g, 4.04 mmol) in DMF (3.5 mL). The resulting mixture was heated to 95 °C for 3 hours, and then cooled to room temperature. The reaction mixture was diluted with ethyl acetate (75 mL), and the organic layer was washed with H2O, then with brine, dried over Na2SO4, filtered, and concentrated. The crude product was used directly for the next step without further purification.

[0375] Step g: Dissolve 0.75 g (3.65 mmol) of 5'-fluorospiro[cyclopentane-1,1'-isoindoline]-3'-one from step f in concentrated H₂SO₄ (5 mL) and cool to 0 °C. Add dropwise 70% HNO₃ (0.46 g, 7.31 mmol, 2.0 equivalence) and stir the reaction mixture at 0 °C for 10 min, then warm to room temperature and stir overnight. Add ice and dilute the mixture with cold water (10 mL). Extract the reaction mixture with EtOAc (2 × 25 mL), wash with H₂O, then wash with brine, dry with Na₂SO₄, filter, and concentrate. The crude product (0.55 g) can be used directly for the next step without further purification. MS: C 12 H 11 FN2O3[M+H] + The calculated value of (ES)m / z is 251.1, and the measured value is 251.0.

[0376] Step h: 5'-fluoro-4'-nitrospiro[cyclopentane-1,1'-isoindoline]-3'-one (0.55 g, 1.32 mmol) and 20 mL of MeOH at 10% Pd / C (50% humidity, 200 mg) were stirred for 1 hour under a hydrogen atmosphere (40 psi). The mixture was filtered through diatomaceous earth and washed with MeOH (40 mL). The filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel chromatography (20-100% ethyl acetate in hexane) to give 4'-amino-5'-fluorospiro[cyclopentane-1,1'-isoindoline]-3'-one (0.45 g, 56%). MS: C 12 H 13 FN2O[M+H] + The calculated value of (ES)m / z is 221.0, and the measured value is also 221.0.

[0377] Step i: At room temperature, N-chlorosuccinimide (89 mg, 0.67 mmol) was added to a stirred solution of 7'-amino-6'-fluoro-spiro[cyclopentane-1,3'-isoindoline]-1'-one (135 mg, 0.61 mmol) in AcOH (1.5 mL). The resulting mixture was heated to 45 °C for 16 hours, then cooled to room temperature. The reaction mixture was diluted with EtOAc (50 mL). The organic layer was washed with H2O, then with a brine solution, dried over Na2SO4, filtered, and concentrated. The crude product was used directly for the next step without further purification.

[0378] Step j: At room temperature, dioxane (0.122 μl, 0.490 mmol) of 4N HCl was added to a mixture of 4'-amino-7'-chloro-5'-fluorospiro[cyclopentan-1,1'-isoindoline]-3'-one (125 mg, 0.490 mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (104 mg, 0.735 mmol) in anhydrous methanol (2 mL). The reaction mixture was stirred at 60 °C for 3 hours and then concentrated. Ethyl acetate (5 mL) was added to the residue and stirred at 50 °C for 10 minutes, then cooled to room temperature. The mixture was filtered and dried to give 3-((7'-chloro-5'-fluoro-3'-oxospiro-[cyclopentan-1,1'-isoindoline]-4'-yl)amino)-4-methoxycyclobut-3-ene-1,2-dione. MS: C 17 H 14 ClFN2O4[M+H] + The calculated value of (ES)m / z is 365.1, and the measured value is 365.0.

[0379] Step k: At room temperature, triethylamine (114 mg, 0.41 mmol, 2.0 mmol) was added to a methanol mixture of 3-((7'-chloro-5'-fluoro-3'-oxospiro[cyclopentane-1,1'-isoindoline]-4'-yl)amino)-4-methoxycyclobut-3-ene-1,2-dione (70 mg, 0.205 mmol) and (1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propane-1-amine (32 mg, 0.205 mmol) from step i. The mixture was stirred at 60 °C for 3 hours and then cooled to room temperature. The solvent was removed under reduced pressure, and the crude compound was purified by silica gel chromatography (hexane in 20-100% ethyl acetate) to give (R)-3-((7'-chloro-5'-fluoro-3'-oxospiro[cyclopentan-1,1'-isoindoline]-4'-yl)amino)-4-((2,2-dimethyl-1-(5-methylfuran-2-yl)propyl)amino)cyclobut-3-en-1,2-dione). 1 H NMR (400MHz, DMSO-d6) δ9.71(s,1H),9.42(s,1H),8.50(d,J=10.2Hz,1H),7.70(d,J=8.6Hz,1H),6.19(d,J=4.2Hz ,1H),6.03-6.01(m,1H),5.02(d,J=10.2Hz,1H),2.27(s,3H),1.95-1.80(m,6H),1.70-1.80(m,2H),0.95(s,9H). MS:C 26 H 27 ClFN3O4[M+H] +The calculated value of (ES)m / z is 500.2, and the measured value is also 500.2.

[0380] Example 21: Synthesis of 3-[(7-chloro-2-hydroxy-3-oxo-isoindoline-4-yl)amino]-4-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propyl]amino]cyclobut-3-ene-1,2-dione

[0381]

[0382] Step a: Triethylamine (983 mg, 9.71 mmol) was added to a mixture of methyl 2-(bromomethyl)-3-chloro-6-nitrobenzoate (1.5 g, 4.87 mmol) and tert-butyl N-hydroxycarbamate (710 mg, 5.35 mmol) in THF (10 mL), and the mixture was heated to 65 °C for 16 hours. After completion, the reaction mixture was diluted with EtOAc and washed with H2O (3x). The organic layer was dried with Na2SO4, filtered, and concentrated to obtain the product, which was immediately used for the next step.

[0383] Step b: Add 24 mL of an 8:2 mixture of EtOH and H₂O to the crude 4-chloro-2-hydroxy-7-nitro-isoindoline-1-one product from the previous step. Add NH₄Cl (2.67 g, 49.9 mmol) and iron powder (800 mg, 14.3 mmol) to the solution. Heat the reaction to 85 °C. Once complete, concentrate the reaction to remove EtOH and add EtOAc and H₂O. Filter the mixture to remove iron, then wash with H₂O, dry with Na₂SO₄, filter and concentrate. Add 2.0 mL of MeOH containing HCl to the crude product, filter to collect the solid, and give the product.

[0384] Step c: A mixture of 3,4-dimethoxycyclobut-3-ene-1,2-dione (80 mg, 0.56 mmol) in anhydrous methanol (2.5 mL) (75 mg, 0.38 mmol) was stirred overnight at 60 °C. The reaction mixture was filtered, the solid was washed with MeOH, and then dried under vacuum to give the crude product.

[0385] Step d: The crude 3-[(7-chloro-2-hydroxy-3-oxo-isoindoline-4-yl)amino]-4-methoxy-cyclobut-3-ene-1,2-dione and (1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propane-1-amine hydrochloride (66 mg, 0.40 mmol) obtained from the previous step were combined in MeOH (2.0 mL) and triethylamine (76 mg, 0.76 mmol) was added. The mixture was stirred overnight at room temperature. The reaction was concentrated and purified sequentially by silica gel chromatography and reversed-phase chromatography (MeCN: H2O containing 0.1% TFA as eluent) to give the product. 1 H NMR (400MHz, DMSO-d6) δ11.76(s,1H),10.42(s,1H),9.19(d,J=9.7Hz,1H),7.65(d,J=9.0Hz,1H),7.3 9(d,J=9.0Hz,1H),6.22-6.18(m,1H),6.05-6.03(m,1H),5.09-5.15(m,3H),2.26(s,3H),0.95(s,9H). MS:C 22 H 22 ClN3O5[M+Na] + The calculated value of (ES)m / z is 464.1, and the measured value is 464.0.

[0386] Example 22: Synthesis of 3-[(2-amino-7-chloro-3-oxo-isoindoline-4-yl)amino]-4-[[(1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propyl]amino]cyclobut-3-ene-1,2-dione

[0387]

[0388] Step a: A mixture of methyl 2-(bromomethyl)-3-chloro-6-nitrobenzoate (1.5 g, 4.87 mmol) and tert-butyl N-carbamate (670 mg, 5.11 mmol) in THF (10 mL) was heated to 65 °C for 3 hours. After completion, the reaction mixture was diluted with EtOAc and washed with H2O (3x). The organic layer was dried over Na2SO4, filtered, and concentrated to obtain the crude product, which was then purified by silica gel chromatography to obtain the final product.

[0389] Step b: Add MeOH (15 mL) to N-(4-chloro-7-nitro-1-oxo-isoindoline-2-yl)carbamate tert-butyl ester (1.6 g, 4.9 mmol) from the previous step. Add PtO2 (221 mg, 0.97 mmol) to the solution, and shake the mixture in a hydrogenation apparatus under H2 (30 psi). After the reaction is complete, filter the mixture and dry it under vacuum to obtain the product.

[0390] Step c: A mixture of N-(7-amino-4-chloro-1-oxo-isoindoline-2-yl)carbamate tert-butyl ester (420 mg, 1.84 mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (392 mg, 2.7 mmol) in anhydrous methanol (5.0 mL) was stirred at 60 °C for 12 hours. The reaction mixture was filtered, and the solid was purified by silica gel chromatography to give the product.

[0391] Step d: N-[4-chloro-7-[(2-methoxy-3,4-dioxo-cyclobuten-1-yl)amino]-1-oxo-isoindoline-2-yl]carbamate tert-butyl (50 mg, 0.12 mmol) and (1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propane-1-amine hydrochloride (27 mg, 0.13 mmol) were combined in MeOH (4.0 mL), and triethylamine (24 mg, 0.244 mmol) was added. The mixture was stirred overnight at 65 °C. The reaction was then concentrated to give a crude product, which was used for the next step without further purification.

[0392] Step e: Dissolve the crude product from the previous step in MeOH (2.0 mL), add dioxane (4 M, 10 drops) containing HCl, and stir the reaction at room temperature. After completion, concentrate the reaction and purify it by reversed-phase chromatography (MeCN: H2O containing 0.1% TFA as eluent) to obtain the product. 1 H NMR (400MHz, DMSO-d6) δ9.87(s,1H),9.01(d,J=10.2Hz,1H),8.05(d,J=6.0Hz,2H),7.43(d,J=9.0Hz,1H),7.23(d,J=9 .0Hz,1H),6.1(d,J=3.2Hz,1H),5.04(d,J=4.2Hz,1H),4.40(d,J=11.2Hz,2H),2.09(s,3H),1.79(s,3H),0.86(s,9H). MS:C 23 H 25 ClN4O4[M+H] + The calculated value of (ES)m / z is 457.2, while the measured value is 457.0.

[0393] Example 23: Synthesis of 3-[(7-chloro-2-methoxy-3-oxo-isoindoline-4-yl)amino]-4-[[(1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propyl]amino]cyclobut-3-ene-1,2-dione

[0394]

[0395] Step a: A mixture of methyl 2-(bromomethyl)-3-chloro-6-nitrobenzoate (5.0 g, 16.2 mmol) and O-methylhydroxylamine hydrochloride (2.12 g, 17.9 mmol) in THF (30 mL) was heated to 65 °C for 2 hours. After completion, the reaction mixture was diluted with EtOAc and washed with H2O (3x). The organic layer was dried over Na2SO4, filtered, and concentrated to give the product, which was used for the next step without further purification.

[0396] Step b: Add 26 mL of an EtOH:H2O mixture in an 8:2 ratio to the product from the previous step. Add NH4Cl (9.0 g, 170 mmol) and iron powder (2.27 g, 40.6 mmol) to this solution and heat the mixture to 85 °C. Once complete, concentrate the reaction mixture to remove EtOH, then add EtOAc and H2O. Filter the mixture to remove iron, then wash with H2O, dry with Na2SO4, filter and concentrate. Purify the crude product to obtain the final product by silica gel chromatography (10% to 80% hexane of EtOAc).

[0397] Step c: A mixture of 7-amino-4-chloro-2-methoxy-isoindoline-1-one (1.0 g, 4.1 mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (0.88 g, 6.2 mmol) in anhydrous methanol (10 mL) was stirred overnight at 60 °C. The mixture was then filtered, and the solid was washed with MeOH and dried under vacuum to give the product.

[0398] Step d: 3-[(7-chloro-2-methoxy-3-oxo-isoindoline-4-yl)amino]-4-methoxy-cyclobut-3-ene-1,2-dione (125 mg, 0.39 mmol) and (1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propane-1-amine hydrochloride were combined in MeOH (4.0 mL), and triethylamine (117 mg, 1.15 mmol) was added. The mixture was stirred overnight at 60 °C. The reactants were then concentrated to give a crude product, which was purified by silica gel chromatography (CH2Cl2:MeOH) to obtain the final product. 1HNMR(400MHz,DMSO-d6)δ9.87(s,1H),9.01(d,J=10.2Hz,1H),7.63(d,J=9.0Hz,1H),7.42(d,J=9.0Hz,1H ), 6.09 (s, 1H), 5.04 (d, J = 10.1Hz, 1H), 4.73 (s, 2H), 3.86 (s, 3H), 2.18 (s, 3H), 1.87 (s, 3H), 0.95 (s, 9H). MS:C 24 H 26 The calculated (ES) m / z value of ClN3O5[MH]- is 470.2, while the measured value is 470.1.

[0399] Example 24: Synthesis of 3-(((R)-1-(4,5-dimethylfuran-2-yl)-2,2-dimethylpropyl)amino)-4-(((S)-5-fluoro-1-methyl-3-oxoisoindololin-4-yl)amino)cyclobut-3-en-1,2-dione and 3-(((R)-1-(4,5-dimethylfuran-2-yl)-2,2-dimethylpropyl)amino)-4-(((R)-5-fluoro-1-methyl-3-oxoisoindololin-4-yl))amino)cyclobut-3-en-1,2-dione

[0400]

[0401] Step a: At room temperature, triethylamine (16.72 mmol, 165.5 mmol, 21.8 mL), di-tert-butyl dicarbonate (17.3 g, 79.4 mmol), and catalytic DMAP (100 mg) were added to anhydrous dichloromethane (100 mL) containing 10 g of 6-fluoroisoindolin-1-one (66.2 mmol) under stirring. The reaction mixture was stirred at room temperature for 16 hours. After completion, it was diluted with CH2Cl2, washed with H2O, and then washed with a saturated aqueous solution of NaHCO3. The organic layer was dried over Na2SO4, filtered, and concentrated under vacuum. The crude compound was purified by silica gel chromatography (0-30% ethyl acetate in hexane) to give the product. MS: C 13 H 14 FNO3[M+H] + The calculated value of (ES)m / z is 252.3, and the measured value is also 252.3.

[0402] Step b: 1) Under a nitrogen atmosphere at -78°C, LiHMDS (21.89 mL, 21.89 mmol) was added dropwise to an anhydrous THF (40 mL) stirred solution of 6-fluoro-1-oxoisoindoline-2-carboxylic acid tert-butyl ester (5.0 g, 19.9 mmol). After stirring for 30 minutes, a solution of iodomethane (2.82 g, 19.92 mmol) in THF (5 mL) was added to the mixture. The reaction mixture was stirred at -78°C for 1 hour, then warmed to room temperature and stirred for 2 hours. After completion, the reaction mixture was quenched with a saturated NH4Cl aqueous solution, diluted with EtOAc (100 mL), and the organic layer was washed with H2O followed by a brine solution. The organic layer was then dried with Na2SO4, filtered, and concentrated under vacuum. The crude product was used directly for the next step without further purification. 2) Dioxane (79.6 mmol, 20 mL) containing 4N HCl was added to a stirred solution of 6.2 g (66.2 mmol) of 5-fluoro-1-methyl-3-oxoisoindoline-2-carboxylic acid tert-butyl ester in MeOH (60 mL). The mixture was stirred at room temperature for 3 hours. After the reaction was complete, the solvent was removed and the reaction mixture was diluted with EtOAc (3 × 50 mL). The mixture was washed with H2O, followed by a saturated aqueous solution of NaHCO3. The organic layer was then dried over Na2SO4, filtered, and concentrated. The crude compound was purified by silica gel chromatography (10–80% ethyl acetate in hexane) to give the product. MS: C9H8FNO[M+H] + The calculated value of (ES)m / z is 166.2, and the measured value is also 166.2.

[0403] Step c: 1) Under a nitrogen atmosphere, n-BuLi (6.64 mL, 16.61 mmol, 2.5 M hexane) was added dropwise to a stirred solution of 6-fluoro-3-methylisoindolin-1-one (2.5 g, 15.1 mmol) in anhydrous THF (25 mL). The reaction mixture was stirred at -78 °C for 30 min. Then, (1R,2S,5R)-2-isopropyl-5-methylcyclohexylcarboxylate (3.96 g, 18.18 mmol) in THF (5 mL) was added to the mixture, and the mixture was stirred at -78 °C for 30 min. The reaction mixture was then warmed to room temperature and stirred for 3 h. After the reaction was complete, the reaction mixture was quenched with a saturated aqueous NH4Cl solution, extracted with EtOAc (2 × 75 mL), and the combined organic layers were washed with H2O, then washed with a brine solution, dried with Na2SO4, filtered under vacuum, and concentrated. The crude compound was purified by silica gel chromatography to obtain (1S)-(1R,2S,5R)-2-isopropyl-5-methylcyclohexyl-5-fluoro-1-methyl-3-oxoisoindoline-2-carboxylate and (1R)-(1R,2S,5R)-2-isopropyl-5-methylcyclohexyl-5-fluoro-1-methyl-3-oxoisoindoline-2-carboxylate, respectively. 2) At room temperature, 17.2 mmol, 10 mL of MeOH containing Mg(OMe)2 (10-12 wt%) was added to a stirred solution of one of the diastereomers obtained above in 10 mL of MeOH. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, the solvent was removed, and the reaction mixture was quenched with saturated NH4Cl aqueous solution and extracted with EtOAc (2 × 75 mL). The combined organic layers were washed with H₂O, then with a brine solution, dried over Na₂SO₄, filtered, and concentrated under vacuum. The crude compound was purified by silica gel chromatography (20-60% ethyl acetate in hexane) to obtain the desired product. MS: C₁₈H₈FNO[M+H] + The calculated (ES)m / z value is 166.2, and the measured value is also 166.2. Another diastereomer was similarly treated to obtain another desired product.

[0404] Step d: 1) Dissolve one of the compounds obtained in step c (0.45 g, 2.72 mmol) in concentrated H₂SO₄ (5 mL) and cool to 0 °C. Add dropwise 70% HNO₃ (0.34 g, 24.1 mmol, 2.0 equivalence) and stir the reaction mixture at 0 °C for 10 minutes, then warm to room temperature and stir overnight. Add ice, then dilute the mixture with cold water (10 mL) and extract with EtOAc (2 × 25 mL). Wash the combined organic layers with H₂O, then with brine, then dry with Na₂SO₄, filter, and concentrate. The crude product can be used directly for the next step without further purification. MS: C₉H₇F₂N₂O₃[M+H] + The calculated (ES) m / z value was 211.0, and the measured value was 211.2. Another enantiomer was similarly treated to obtain another desired product. 2) Under a hydrogen atmosphere (40 psi), one of the above-obtained compounds (0.35 g, 1.32 mmol) and 10% Pd / C (50% humidity, 100 mg) were stirred in a MeOH (25 mL) solution for 1 hour. The mixture was filtered through diatomaceous earth and washed with MeOH (40 mL). The filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel chromatography (20-100% ethyl acetate / hexane) to obtain the desired product. MS: C9H9FN2O[M+H] + The calculated (ES)m / z value is 181.1, and the measured value is 181.2. Another enantiomer was similarly treated to obtain another desired product.

[0405] Step e: A mixture of one of the compounds obtained in step d (170 mg, 0.939 mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (200 mg, 1.40 mmol) in anhydrous methanol (4 mL) was stirred for 3 hours at 60 °C. The reaction mixture was evaporated, and the residue was stirred in ethyl acetate (10 mL) at 50 °C for 30 minutes, then cooled to room temperature. The mixture was filtered and dried to give the desired product. MS: C 14 H 11 FN₂O₄[M+H] + The calculated (ES)m / z value is 291.1, and the measured value is 291.2. Another enantiomer was similarly treated to obtain another desired product.

[0406] Step f: Triethylamine (75 mg, 0.687 mmol, 2.5 equivalents) was added to a methanol (2.5 mL) solution of (1R)-1-(4,5-dimethyl-2-furanyl)-2,2-dimethyl-propane-1-amine hydrochloride (62 mg, 0.288 mmol, 1.05 equivalents). The mixture was stirred at room temperature for 10 minutes to form a clear solution, and then one of the compounds obtained above was added at room temperature. The resulting solution was stirred at 60 °C for 3 hours. After completion, the reaction was cooled to room temperature. The solvent was removed under reduced pressure, and the crude product was purified by preparative HPLC (acetonitrile-water containing 0.1% TFA) to obtain the desired product. 1 H NMR (400MHz, DMSO-d6) δ9.64(s,1H),8.79(s,1H),8.38(d,J=10.2Hz,1H),7.46(dd,J=11.4,8.2Hz,1H),7.34(dd,J=3.9,8.2Hz ,1H),6.07(s,1H),4.95(d,J=10.1Hz,1H),4.58(q,J=6.6Hz,1H),2.18(s,3H),1.87(s,3H),1.33(d,J=6.6Hz,3H),0.95(s,9H). MS:C 24 H 26 The calculated (ES) m / z value for FN3O4[MH]- is 438.2, while the measured value is 438.0. The acquisition of the other diastereomer was similar. 1 H NMR (400MHz, DMSO-d6) δ9.64(s,1H),8.79(s,1H),8.37(d,J=10.2Hz,1H),7.44(dd,J=11.0,8.2Hz,1H),7.33(dd,J=3.9,8.8Hz ,1H),6.07(s,1H),4.95(d,J=10.1Hz,1H),4.56(q,J=6.3Hz,1H),2.18(s,3H),1.87(s,3H),1.33(d,J=6.6Hz,3H),0.95(s,9H). MS:C 24 H 26 The calculated (ES) m / z value of FN3O4[MH] is 438.2, while the measured value is 438.0.

[0407] Example 25: 3-(((R)-1-(4,5-dimethylfuran-2-yl)-2,2-dimethylbutyl)amino)-4-(((S)-5-fluoro-1,7-dimethyl-3-oxoisoindololin-4-yl)amino)cyclobut-3-ene-1,2-dione and 3-(((R)-1-(4,5-dimethylfuran-2-yl)-2,2-dimethylbutyl)amino)-4-(((R)-5-fluoro-1,7-dimethyl-3-oxoisoindololin-4-yl)amino)cyclobut-3-ene-1,2-dione

[0408]

[0409] Step a: At room temperature, N-iodosuccinimide (350 mg, 1.56 mmol) was added fractionally to a room-temperature water bath solution of AcOH (2.2 mL) of an enantiomeric form of 7-amino-6-fluoro-3-methylisoindolin-1-one (200 mg, 1.11 mmol). The resulting mixture was stirred in a water bath for 30 minutes, quenched with water (1 mL), and then extracted with ethyl acetate (10 mL). The organic layer was washed with brine (10 mL), dried over MgSO4, filtered, and concentrated. The crude product was purified by silica gel chromatography (0-60% ethyl acetate in hexane) to obtain the desired product. MS: C9H8FIN2O[M+H] + The calculated (ES) m / z value is 307.0, and the measured value is also 307.0. Another enantiomer was similarly treated to obtain another desired product.

[0410] Step b: To a solution of one of the compounds obtained in step a (248 mg, 0.81 mmol) in dioxane (8.1 mL), CsF (493 mg, 3.24 mmol), 2,4,6-trimethyl-1,3,5,2,4,6-trioxaborane (305 mg, 2.43 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]-palladium(II) dichloride (66 mg, 0.08 mmol) were added. The resulting mixture was stirred overnight at 80 °C. The reaction mixture was partitioned between water (20 mL) and ethyl acetate (30 mL). The organic layer was washed with brine (20 mL), dried over MgSO4, filtered, and concentrated. The crude product was purified by silica gel chromatography (0-80% ethyl acetate in hexane) to give the desired product. MS: C 10 H 11 FN2O[M+H] + The calculated (ES) m / z value is 195.1, and the measured value is also 195.1. Another enantiomer was similarly treated to obtain another desired product.

[0411] Step c: A mixture of one compound obtained from step b (127 mg, 0.65 mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (186 mg, 1.3 mmol) in anhydrous methanol (3 mL) was stirred overnight at 60 °C, followed by stirring at 80 °C for 5 hours. The reaction mixture was concentrated and purified by silica gel chromatography (0-100% ethyl acetate in hexane) to give the desired product. MS: C 15 H 13 FN₂O₄[M+H] + The calculated (ES) m / z value is 305.1, and the measured value is also 305.1. Another enantiomer was similarly treated to obtain another desired product.

[0412] Step d: Anhydrous methanol (2 mL) was added to a mixture of one compound obtained in step c (40 mg, 0.13 mmol) and (R)-1-(4,5-dimethylfuran-2-yl)-2,2-dimethylbutyl-1-amine (33.6 mg, 0.145 mmol), and the mixture was stirred overnight at 60 °C. The reactants were cooled to room temperature, dissolved in a minimal amount of dichloromethane, and adsorbed onto silica gel. The mixture was purified by silica gel chromatography (40% ethyl acetate in dichloromethane) to obtain the desired product. 1 H NMR (400MHz, DMSO-d6) δ9.55(s,1H),8.78(s,1H),8.32(d,J=10.0Hz,1H),7.29(d,J=11.6Hz,1H),6.06(s,1H),5.05(d,J=10.4Hz,1H),4.63(q,J= 6.8Hz,1H),2.31(s,3H),2.18(s,3H),1.87(s,3H),1.34(d,J=6.8Hz,3H) ,1.26(q,J=7.2Hz,2H),0.93(s,3H),0.88(s,3H),0.82(t,J=7.2Hz,3H). MS:C 26 H 30 The calculated (ES) m / z value for FN3O4[MH]- is 468.2, and the measured value is also 468.2. The acquisition of the other diastereomer was similar.

[0413] Example 26: 3-(((R)-1-(4,5-dimethylfuran-2-yl)-2,2-dimethylpropyl)amino)-4-(((S)-5-fluoro-1,7-dimethyl-3-oxoisoindololin-4-yl)amino)cyclobut-3-ene-1,2-dione and 3-(((R)-1-(4,5-dimethylfuran-2-yl)-2,2-dimethylpropyl)amino)-4-(((R)-5-fluoro-1,7-dimethyl-3-oxoisoindololin-4-yl)amino)cyclobut-3-ene-1,2-dione

[0414]

[0415] Anhydrous methanol (2 mL) was added to a mixture of an enantiomeric form of 3-((5-fluoro-1,7-dimethyl-3-oxoisoindoline-4-yl)amino)-4-methoxycyclobut-3-ene-1,2-dione (40 mg, 0.13 mmol) and (R)-1-(4,5-dimethylfuran-2-yl)-2,2-dimethylbutyl-1-amine (31.6 mg, 0.145 mmol), and the mixture was stirred overnight at 60 °C. The reaction mixture was cooled to room temperature, dissolved in a minimal amount of dichloromethane, and adsorbed onto silica gel. It was purified by silica gel chromatography (40% ethyl acetate in dichloromethane) to give the desired product. 1 H NMR (400MHz, DMSO-d6) δ9.55(s,1H),8.74(s,1H),8.34(d,J=10.4Hz,1H),7.28(d,J=11.6Hz,1H),6.06(s,1H),4.95 (d, J=10.0Hz, 1H), 4.61 (q, J=6.8Hz, 1H), 2.31 (s, 3H), 2.16 (s, 3H), 1.86 (s, 3H), 1.33 (d, J= 6.8Hz, 3H), 0.94 (s, 9H). MS:C 25 H 28 FN3O4[MH] - The calculated (ES)m / z value is 454.2, and the measured value is also 454.2. The other diastereomer was obtained in a similar manner.

[0416] Example 27: Synthesis of 3-(((R)-2,2-dimethyl-1-(5-methylfuran-2-yl)propyl)amino)-4-(((S)-5-fluoro-1-methyl-3-oxoisoindololin-4-yl)amino)cyclobut-3-en-1,2-dione and 3-(((R)-2,2-dimethyl-1-(5-methylfuran-2-yl)propyl)amino)-4-(((R)-5-fluoro-1-methyl-3-oxoisoindololin-4-yl)amino)cyclobut-3-en-1,2-dione

[0417]

[0418] At room temperature, triethylamine (43 mg, 0.42 mmol, 2.5 equivalences) was added to a mixture of an enantiomeric form of 3-((5-fluoro-1-methyl-3-oxoisoindoline-4-yl)amino)-4-methoxycyclobut-3-en-1,2-dione (50 mg, 0.171 mmol) and (1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)propane-1-amine (30 mg, 0.180 mmol) in methanol (3.0 mL). The mixture was stirred at 60 °C for 3 hours and then cooled to room temperature. The solvent was removed under reduced pressure, and the crude product was purified by preparative HPLC (acetonitrile-0.1% TFA in water) to obtain the desired product. 1 H NMR (400MHz, DMSO-d6) δ9.65(s,1H),8.80(s,1H),8.42(d,J=10.1Hz,1H),7.46(dd,J=3.1,8.4Hz,1H),7.36(dd,J=3.9,8.2Hz,1H),6.18 (d,J=3.2,Hz,1H),6.03(d,J=2.4Hz,1H),5.00(d,J=10.5Hz,1H),4.56(q,J=7.1Hz,1H),2.27(s,3H),1.33(d,J=6.6Hz,3H),0.95(s,9H). MS:C 23 H 24 The calculated (ES) m / z value for FN3O4[MH]- is 424.2, while the measured value is 424.0. The other diastereomer is similar. 1 H NMR (400MHz, DMSO-d6) δ9.64(s,1H),8.79(s,1H),8.40(d,J=10.5Hz,1H),7.46(dd,J=11.3,8.6Hz,1H),7.34(dd,J=3.9,8.2Hz,1H),6. 17(d,J=3.2,Hz,1H),6.04-6.03(m,1H),5.00(d,J=10.1Hz,1H),4.56(q,J=6.6Hz,1H),2.27(s,3H),1.33(d,J=6.6Hz,3H),0.95(s,9H). MS:C 23 H 24 FN3O4[M+H] + The calculated value of (ES)m / z is 426.2, and the measured value is 426.0.

[0419] Example 28: Synthesis of 3-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)butyl]amino]-4-[[(1S)-5-fluoro-1-methyl-3-oxo-isoindoline-4-yl]amino]cyclobut-3-ene-1,2-dione and 3-[[(1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)butyl]amino]-4-[[(1R)-5-fluoro-1-methyl-3-oxo-isoindoline-4-yl]amino]cyclobut-3-ene-1,2-dione

[0420]

[0421] Triethylamine (0.025 mL, 0.18 mmol) was added to a mixture of an enantiomeric form of 3-((5-fluoro-1-methyl-3-oxoisoindoline-4-yl)amino)-4-methoxycyclobut-3-ene-1,2-dione (26 mg, 0.09 mmol) and (1R)-2,2-dimethyl-1-(5-methyl-2-furanyl)butane-1-amine hydrochloride (20 mg, 0.09 mmol) in MeOH (1.0 mL). The reaction was stirred at room temperature for 18 hours. Silica gel was then added to the reaction mixture, which was concentrated and purified by silica gel chromatography (CH2Cl2 containing 1% to 10% MeOH). The product was then purified by reversed-phase chromatography (MeCN: H2O containing 0.1% TFA as eluent) to give the final product. 1 H NMR (400MHz, DMSO-d6) δ9.65 (s, 1H), 8.79 (s, 1H), 8.40 (d, J = 10.2Hz, 1H), 7.47 (d d,J=11.1,8.2Hz,1H),7.35(dd,J=8.3,3.8Hz,1H),6.17(d,J=3.1Hz,1H),6.06–6 .01(m,1H),5.10(d,J=10.2Hz,1H),4.58(q,J=6.6Hz,1H),2.27(s,3H),1.34(d,J =6.7Hz,3H),1.32–1.24(m,2H),0.94(s,3H),0.88(s,3H),0.83(t,J=7.4Hz,3H). MS:C 24 H 27 FN3O4[M+H] + The calculated (ES)m / z value is 440.2, and the measured value is 440.4. The other diastereomer is similar.

[0422] The following compounds were prepared using similar synthetic methods and appropriate reagents as described herein, and characterized by MS (mass spectrometry) and / or NMR as shown in Table 1.

[0423] Table 1. Characterization of compounds

[0424]

[0425]

[0426]

[0427]

[0428]

[0429]

[0430]

[0431]

[0432]

[0433]

[0434]

[0435]

[0436]

[0437]

[0438]

[0439]

[0440]

[0441]

[0442]

[0443]

[0444]

[0445]

[0446]

[0447]

[0448]

[0449] Bioactivity

[0450] Biological Example 1: CXCR2 Active Ligand Binding Assay

[0451] Ligand binding assays were used to determine the ability of potential CXCR2 antagonists to block the interaction between CXCR2 and any of its ligands. HEK-293 cells stably expressing CXCR2 or human neutrophils expressing CXCR2 were centrifuged in assay buffer (20 mM HEPES pH 7.1, 140 mM NaCl, 1 mM CaCl2, 5 mM MgCl2, 0.1% sodium azide, and 0.1% bovine serum albumin) and resuspended to 5 × 10⁻⁶. 5 The concentration was [number of cells / mL]. The assay setup was as follows: the compounds used for screening were serially diluted from a maximum of 20 μM, and 0.1 mL containing 5 x 10 [units of concentration] were [dissolved / concentrated]. 4 1 cell (for HEK-293 cells) or 3 x 10 4 One cell line (for human neutrophils) was added to each well containing the compound. Then, 0.1 mL of the compound was added and diluted in assay buffer to the specified concentration. The final concentration, producing / hole 125 I-labeled CXCL8 (obtained from PerkinElmer; Waltham, MAS, USA) plates were sealed and incubated on a shaker at 25°C for approximately 3 hours. Reactants were aspirated onto GF / B glass filters pre-soaked in 0.3% polyethyleneimine (PEI) solution using a vacuum cell collector (Packard Instruments; Meridon, Connecticut). Scintillation solution (50 μL; Microscint 20, Packard Instruments) was added to each well, the plates were sealed, and radioactivity was measured using a Top Count scintillation counter (Packard Instruments). Control wells containing only the diluent (total count) or 20 μM of the compound were used to calculate the total percentage of inhibition (IC). The Prism computer program from GraphPad Inc. (San Diego, CA) was used to calculate IC. 50 Value. IC 50 The value represents the concentration required to reduce the binding of labeled CXCR8 to the receptor by 50%. In Figure 1, during the binding assay, an IC50 concentration of less than 100 nM is used. 50 Compounds with a value of 100-1000 nM are labeled as (++); compounds with a value of 20 μM or less but higher than 1000 nM are labeled as (++); and compounds with a value of 20 μM or less but higher than 1000 nM are labeled as (+).

[0452] Biological Example 2: Migratory / Chemotropic Assay of CXCR2 Activity

[0453] Serum chemotaxis assays can be used to determine the efficacy of potential receptor antagonists in blocking migration mediated by chemokine receptors such as CXCR2. This assay typically uses a polycarbonate membrane with a 5 μm pore size. The microcompartment system was used. The assay began by collecting cells expressing chemokine receptors (neutrophils isolated from human whole blood in this example) by centrifugation at 400×g at room temperature, followed by resuscitation in human serum at 4 million / ml. The analyte was serially diluted from a maximum final concentration of 10 μM (or an equivalent volume of solvent (DMSO)) and then added to the cell / serum mixture. Separately, the analyte was analyzed at its EC... 50 Recombinant human CXCL5 (ENA-78) at a concentration (10 nM) was placed In the lower wells of the plate, place a 5-μm (pore size) polycarbonate membrane on the plate and transfer 20 μL of the cell / compound mixture to each well. Incubate the plate at 37°C for 45 minutes, then remove the polycarbonate membrane and add 5 μL of DNA inserter CyQUANT (Ingenieur, Carlsbad, California) to the lower wells. Measure the fluorescence intensity corresponding to the number of migrating cells using a Spectrafluor Plus plate reader (Teken, San Jose, California).

[0454] Biological Example 3: Migratory / Chemotropic Assay of CCR6 Activity

[0455] Serum chemotaxis assays are used to determine the efficacy of potential receptor antagonists in blocking migration mediated by chemokine receptors, such as CCR6. This assay typically uses a polycarbonate membrane with a 5 μm pore size. The assay was performed using a microchamber system. The assay began by collecting cells expressing chemokine receptors (KHYG-1 cells in this example, Yagida et al., Leukemia, 14:922, 2000) by centrifugation at 400 × g at room temperature, and then suspending them in human serum at 4 million / ml. The analyte was serially diluted from a maximum final concentration of 10 μM (or its equivalent volume in solvent (DMSO)) and then added to the cell / serum mixture. Separately, the analyte was analyzed on its EC... 50 Recombinant human CCL20 (MIP-3α / LARC) at a concentration (10 nM) was placed In the lower wells of the plate, a 5-μm (pore size) polycarbonate membrane was placed on the plate, and 20 μL of the cell / compound mixture was transferred to each well of the membrane. The plate was incubated at 37°C for 45 min, then the polycarbonate membrane was removed, and 5 μL of DNA inserter CyQUANT (Ingenieur, Carlsbad, California) was added to the lower wells. The fluorescence intensity corresponding to the number of migrating cells was measured using a SpectrafluorPlus plate reader (Teken, San Jose, California). In Figure 1, in chemotaxis assays, an IC50 value of less than 100 nM is observed. 50 Compounds with a value of 100-1000 nM are labeled as (++); those with a value of 20 μM or less but higher than 1000 nM are labeled as (+).

[0456] Biological Example 4: In vivo efficacy in an IL-23-induced psoriatic ear swelling model

[0457] Intradermal injection of IL-23 into the ear of mice induced CCR6-dependent ear swelling (Hendrick May et al., Journal of Clinical Investigations, 2009, 119: 2317-2329). C57Bl / 6 mice were injected intradermally with IL-23 in the right ear, and PBS was injected intradermally in the left ear as a control. Compound 1.023 (synthesized in Example 6) was administered subcutaneously. After three intradermal injections of IL-23 and the onset of moderate ear swelling, the compound was administered therapeutically. The degree of swelling was measured using calipers. Compound 1.023 completely inhibited IL-23-induced ear swelling and reduced swelling to baseline levels. Figure 2 ).

[0458] Biological Example 5: In vivo efficacy in an imiquimod-induced psoriasis-like model

[0459] Topical application of imiquimod cream to the shaved backs of mice resulted in the development of psoriatic lesions resembling those in humans, namely erythema, skin thickness, and desquamation. (Van der Fitz L. et al., 2009. Journal of Immunology 182: 5836-5845). Balb / c mice were treated with topical imiquimod cream on the shaved backs. Compound 1.129 was prophylactically administered orally to achieve adequate plasma concentrations throughout the study. The development of psoriatic lesions was determined in a blinded manner by measuring three aspects of skin disease: erythema severity, percentage of skin affected by desquamation, and skin thickness measured by calipers. Each measurement was assigned a disease score between 0 (no disease) and 4 (maximum disease) to calculate a cumulative PASI (Psoriasis Activity Severity Index) score, with a maximum score of 12. Compound 1.129 was able to reduce the severity of the cumulative PASI score by inhibiting erythema, desquamation, and skin thickness. Compared with vector-treated mice ( Figure 3 In the group treated with compound 1.129, the percentage of mice exhibiting severe symptoms (score ≥3 for each reading) was reduced.

[0460] Specific embodiments of the invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations of the disclosed embodiments may become apparent to those skilled in the art upon reading the foregoing description, and such variations are expected to be suitably adopted by those skilled in the art. Therefore, the invention is intended to be practiced in ways other than those specifically described herein, and the invention includes all modifications and equivalents of the subject matter described in the appended claims as permitted by applicable law. Furthermore, unless otherwise stated herein or clearly contradicted by the context, the invention covers any combination of the foregoing elements in all possible variations.

[0461] All publications, patent applications, accession numbers and other references cited in this specification are incorporated herein by reference as if each individual publication or patent application were specifically and individually indicated as incorporated by reference.

Claims

1. A compound having formula (A), or a pharmaceutically acceptable salt, tautomer, or rotational isomer thereof, （A） in, Selected from the following group: ; B is selected from the following group: ; R 3 They are members selected from H and D; R 4 It is selected from H and C 1-3 Alkyl or Y, wherein C 1-3 The alkyl group is replaced by a tetrazolonyl group, wherein the tetrazolonyl group is selected from C1 or C2 by 0 or 1. 1-6 Alkyl, C 1-6 Hydroxyalkyl or C 1-4 Alkyl-OC 1-4 Alkyl group substitution, wherein Y is selected from the group consisting of pyridyl, pyrazolyl, and phenyl, wherein pyridyl, pyrazolyl, and phenyl have 1-3 independently selected halogens, -C 1-4 Alkyl, -C 1-4 Alkyl groups and -CO2H substituents; R 5a and R 5b Each element is independently selected from H, halogen, and C. 1-4 Alkyl, -C 1-4 Halogenated alkyl, OC 1-4 Haloalkyl, C 1-4 Members of alkoxy, CO2H, and CN; R 7 It is selected from methyl, ethyl and C 1-2 Members of haloalkyl groups.

2. The compound of claim 1, or a pharmaceutically acceptable salt, tautomer, or rotational isomer thereof, wherein B is selected from the group consisting of: 。 3. The compound of claim 1, or a pharmaceutically acceptable salt, tautomer, or rotational isomer thereof, wherein R 3 For H.

4. The compound of claim 1, or a pharmaceutically acceptable salt, tautomer, or rotational isomer thereof, wherein each R 5a and R 5b Selected independently from the following groups: H, CH3, Cl, and F.

5. The compound of claim 1, or a pharmaceutically acceptable salt, tautomer, or rotational isomer thereof, wherein... Selected from the following group: 。 6. The compound of claim 1, or a pharmaceutically acceptable salt, tautomer, or rotational isomer thereof, wherein... Selected from the following group: 。 7. The compound of claim 1, or a pharmaceutically acceptable salt, tautomer, or rotational isomer thereof, wherein R 6a and R 6b Each is independently selected from H and C 1-2 alkyl.

8. The compound of claim 1, or a pharmaceutically acceptable salt, tautomer, or rotational isomer thereof, wherein R 4 Selected from the following group: 。 9. The compound of claim 1, or a pharmaceutically acceptable salt, tautomer or rotational isomer thereof, wherein R7 is selected from the group consisting of methyl, ethyl and CF3.

10. The compound of claim 1, or a pharmaceutically acceptable salt, tautomer, or rotational isomer thereof, wherein the compound carries R 3 The carbon atoms are essentially free of other isomers.

11. The compound of claim 1, or a pharmaceutically acceptable salt, tautomer, or rotational isomer thereof, wherein R 4 It's Y.

12. The compound of claim 1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of: and .

13. The compound of claim 1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of: 。 14. The compound of claim 1, or a pharmaceutically acceptable salt thereof, having the following structure: 。 15. The compound of claim 1, or a pharmaceutically acceptable salt thereof, having the following structure: 。 16. The compound of claim 1, or a pharmaceutically acceptable salt thereof, having the following structure: 。 17. The compound of claim 1, or a pharmaceutically acceptable salt thereof, having the following structure: 。 18. The compound of claim 1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of: 。 19. A pharmaceutical composition comprising a compound of any one of claims 1 to 18.

20. Use of a compound of any one of claims 1 to 18, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 19, characterized in that, This is used to prepare a drug for treating CXCR2 and / or CCR6-mediated diseases or conditions, wherein the CXCR2 and / or CCR6-mediated diseases or conditions are psoriasis.

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