2-Substituted 3,4a,5,7,8,8a-hexahydro-4H-thiopyrano[4,3-d]pyrimidin-4-ones for wound treatment

Abstract

Provided herein are compounds, pharmaceutical compositions containing the compounds, methods for making the compounds, and methods for using the compounds and compositions.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is a PCT international application claiming the benefit of U.S. Provisional Application No. 63 / 417,257, filed October 18, 2022, U.S. Provisional Application No. 63 / 418,947, filed October 24, 2022, and U.S. Provisional Application No. 63 / 418,956, filed October 24, 2022, the contents of each of which are incorporated herein by reference in their entirety.

[0002] Field Provided herein are compounds, pharmaceutical compositions containing the compounds, methods for making the compounds, and methods for using the compounds and compositions in treating wounds, particularly enhancing tissue regeneration after wound healing. Also provided herein are methods for treating a condition associated with Wnt transcripts or Wnt signaling pathway activity in a mammal, comprising administering a therapeutically effective amount of the compound or composition to the mammal. [Background technology]

[0003] background The Wnt pathway has been shown to play an important role in skin fibrosis and scarring. The Wnt pathway is an evolutionarily conserved pathway that controls important aspects of cell fate determination, cell polarity, cell migration, neural patterning, and organogenesis during embryonic development. This pathway contributes to proper tissue development in embryos and tissue maintenance in adults. Wnt signaling is involved in the early stages of skin development, where, after blastocyst formation, ectodermal and mesodermal germ cells differentiate to form the epidermis and dermis, respectively.

[0004] There are at least three distinct Wnt signaling pathways, but the canonical (or β-catenin-dependent) Wnt pathway is the most well-understood. β-catenin is the primary effector molecule resulting from canonical Wnt pathway signaling, and its protein levels are regulated via the "destruction complex." In the absence of Wnt signaling, the transcriptional activator β-catenin is actively degraded within the cell by the action of a protein complex called the "destruction complex." Within this complex, Axin-1 and -2 form a scaffold with adenomatous polypsis coli, which promotes the phosphorylation of β-catenin by casein kinase 19a and glycogen synthase kinase 3β. The phosphorylated β-catenin is recognized, ubiquitinated, and undergoes proteosomal degradation. Tankryases I and II (TK1 and TK2) are poly(ADP-ribose) polymerases (PARPs) that parsylate and destabilize Axin-1 and -2 proteins, destabilizing the β-catenin degradation complex. Shortly after the degradation complex is destabilized, β-catenin is dephosphorylated and subsequently stabilized, accumulating in the cytoplasm and entering the nucleus, where it interacts with members of the Tcf / Lef family. β-catenin converts Tcf proteins into potent transcriptional activators by recruiting coactivator proteins, ensuring efficient activation of Wnt target genes. Shortly after activation by the Wnt family of natural ligands, the Wnt pathway upregulates TNK1 and 2, which help destabilize the degradation complex. Studies have shown that TNK1 and 2 are key regulators of canonical Wnt signaling.

[0005] XAV939 demonstrated an IC 50 Selectively inhibits Wnt / β-catenin-mediated transcription through TK1 and TK2 inhibition at 11 nM / 4 nM, modulating axin levels and CRE, F- KIt is a small molecule that does not affect TGF-β or TGF-β. Recently, topical application of XAV939 in a mouse ear punch assay demonstrated that XAV939 significantly increased wound closure rates while reducing fibrosis (scarring) (Bastakoty, D. et al. FASEB J., 2015, 29(12): 4881-4892). However, XAV939 was only used as a "research tool" compound because it is soluble in dimethyl sulfoxide (DMSO) and has very low water solubility (<1 μg / mL). The problem with this approach is that humans cannot tolerate DMSO.

[0006] Matrix components including graphene oxide (GO) and hyaluronic acid (HA) have been shown to be useful in providing a support matrix for XAV939, enabling its use as a therapeutic agent for wound healing in humans and animals. See, e.g., US20210000959, where XAV939 in a GO-HA matrix substantially improves the quality of wound healing; specifically, it induces tissue to limit scarring, following the fibrotic wound healing pathway. Furthermore, increased cartilage regeneration and healing after acute injury has been observed, for example, in 2 mm biopsy punch wounds made in the center of the cartilage region of XAV939-treated C57Bl / CJ mice (Bastakoty, D. et al. FASEB J., 2015, 29(12): 4881-4892 and WO2023 / 039298).

[0007] Although the therapeutic utility of XAV939 has been demonstrated, XAV939 has poor solubility in aqueous solutions. Thus, there is a need for Wnt inhibitors with improved / different properties, such as increased aqueous solubility, increased Wnt inhibition, increased bioavailability, and / or modified biological stability. Summary of the Invention

[0008] overview In a first aspect, provided herein is a compound of formula (I), or any prodrug, pharmaceutically acceptable salt, metabolite, polymorph, solvate, hydrate, stereoisomer, and / or tautomer thereof:

[0009] [ka]

[0010] (In the formula, R 1 is hydrogen, deuterium, C1-C3 alkyl, —OH, —O—C1-C3 alkyl, —CH2OH, or —B(OH)2; R 1a is hydrogen, deuterium, or C1-C3 alkyl; R 2 teeth, (a)R 3 and optionally 1, 2 or 3 R 3a phenyl optionally substituted by a group; (b) phenyl substituted with a 5- or 6-membered heteroaryl, wherein the 5- or 6-membered heteroaryl is R 3 and optionally 1, 2 or 3 R 3a groups, and the phenyl may further optionally be substituted with one, two or three R 3a optionally substituted with a group, (c) optionally 1, 2, or 3 R 3a and may be substituted with -phenyl-R 3 phenyl substituted with -phenyl-R 3 The phenyl in the group may optionally be substituted with one, two or three R 3a optionally substituted with a group); (d)R 3 and optionally 1, 2 or 3 R 3a 5- or 6-membered heteroaryl optionally substituted by a group; (e) optionally 1, 2, or 3 R 3a and may be substituted with -phenyl-R 35- or 6-membered heteroaryl substituted with 1, 2, or 3 R 3a optionally substituted with a group); (f) optionally 1, 2, or 3 R 3a and may be substituted with a -(5- or 6-membered heteroaryl)-R 3 5- or 6-membered heteroaryl substituted with -(5- or 6-membered heteroaryl)-R 3 The 5- or 6-membered heteroaryl in 3a optionally substituted with a group); (g)R 3 and optionally one or two R 3a C3-C6 cycloalkyl optionally substituted by a group; (h) C-C cycloalkyl substituted with 5- or 6-membered heteroaryl, wherein the 5- or 6-membered heteroaryl is R 3 and optionally 1, 2 or 3 R 3a groups, and the cycloalkyl may optionally be substituted with one or two R 3a optionally substituted with a group); (i) C-C cycloalkyl substituted with phenyl, wherein the phenyl is R 3 and optionally 1, 2 or 3 R 3a groups, and the cycloalkyl may optionally be substituted with one or two R 3a optionally substituted with a group); (j) 3- to 8-membered heterocycloalkyl substituted with phenyl or 5- or 6-membered heteroaryl, wherein the phenyl and the 5- to 6-membered heteroaryl are R 3 and optionally 1, 2 or 3 R 3a optionally substituted with a group); (k) -CH=CH-R 5 (where R 5is phenyl or 5- or 6-membered heteroaryl, and said phenyl and said 5- or 6-membered heteroaryl are R 3 and optionally 1, 2 or 3 R 3a (which may be substituted with a group) and; R 3 are independently -B(OH)2, cyano, halo, halo-C1-C6 alkyl, -(C0-C6 alkylene)-OR 4 or a 5- to 10-membered heterocycle, wherein the 5- to 10-membered heterocycle is optionally substituted with cyano; or R 2 When (a) is true, R 3 and one R 3a When on adjacent carbons, they are combined with the carbon to which they are attached.

[0011] [ka]

[0012] where the * indicates a carbon atom shared with the phenyl ring, and any remaining R 3a is as defined below, and each R 7a are independently hydrogen or C1-C6 alkyl Forming; Each R 3a is independently selected from cyano, halo, —OH, C-C alkyl, halo-C-C alkyl, and C-C alkoxy; R 4 is hydroxy-C1-C6 alkyl, C1-C6 alkoxy-C1-C6 alkyl, or C1-C6 alkoxycarbonyl-NH-C1-C6 alkyl); (However, the compound is 2-(4-(trifluoromethyl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(4-(4-methoxyphenyl)piperazin-1-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(4-(3-methoxyphenyl)piperazin-1-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(4-(2-methoxyphenyl)piperazin-1-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(4-chlorophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(5-(trifluoromethyl)pyridin-2-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(3-(trifluoromethyl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(3-(trifluoromethyl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(5-chlorothiophen-3-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof. nor is it); In some embodiments, provided is a compound of Formula (I) (or any embodiment thereof), or a single stereoisomer or mixture of stereoisomers thereof; a single tautomer or mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof.

[0013] In a second aspect, provided herein is a pharmaceutical composition comprising a compound of Formula (I) (or any embodiment thereof), or any prodrug, pharmaceutically acceptable salt, metabolite, polymorph, solvate, hydrate, stereoisomer, and / or tautomer thereof; and a pharmaceutically acceptable carrier. In one or more embodiments, provided is a compound of Formula (I) (or any embodiment thereof), or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

[0014] In a third aspect, there is provided a method of inhibiting Wnt transcription or Wnt signaling pathway activity in a subject, comprising contacting an effective amount of a compound of Formula (I) (or any embodiment thereof), or any prodrug, pharmaceutically acceptable salt, metabolite, polymorph, solvate, hydrate, stereoisomer, and / or tautomer thereof. In some embodiments, the compound of Formula (I) (or any embodiment thereof) is provided as a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof.

[0015] In a fourth aspect, provided herein are methods for treating a disease, disorder, or condition associated with Wnt transcription or Wnt signaling pathway activity in a mammal in need thereof, comprising administering a compound of Formula (I) (or any embodiment thereof) or a pharmaceutical composition comprising a compound of Formula (I), or any prodrug, pharmaceutically acceptable salt, metabolite, polymorph, solvate, hydrate, stereoisomer, and / or tautomer thereof. In some embodiments, the compound of Formula (I) (or any embodiment thereof) is provided as a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof. In some embodiments, the method is for stimulating tissue regeneration at a wound in a mammal in need thereof.

[0016] In a fifth aspect, there is provided a method of inducing bacteriostasis associated with Wnt transcription or Wnt signaling pathway activity in a mammal in need thereof, comprising administering to a mammal in need thereof XAV939 or a tautomer thereof and / or a pharmaceutically acceptable salt thereof, optionally as a pharmaceutical composition thereof; administering a compound of formula (I) (or any embodiment thereof), or a single stereoisomer or mixture of stereoisomers thereof, single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof; or administering a pharmaceutical composition of the second aspect (or any embodiment thereof).

[0017] In a sixth aspect, the compound of formula:

[0018] [ka]

[0019] (In the formula, LG 1 is a leaving group such as fluoro, chloro, bromo, iodo, triflate, mesylate, triazole, pyrazole, boronic acid, boronic ester, or allyltrifluoroborate; R 1 is hydrogen, deuterium, C1-C3 alkyl, —OH, —O—C1-C3 alkyl, —CH2OH, or —B(OH)2; R 1a is hydrogen, deuterium, or C1-C3 alkyl; R 20 is alkyl, preferably methyl or ethyl, or CD3; R 2 'teeth, (b1) phenyl substituted with 5- or 6-membered heteroaryl, wherein the 5- or 6-membered heteroaryl is LG 1 and optionally 1, 2 or 3 R 3a groups, and the phenyl may further optionally be substituted with one, two or three R 3a optionally substituted with a group, (c1) optionally 1, 2 or 3 R 3a and may be substituted with -phenyl-LG 1 phenyl substituted with -phenyl-LG 1 The phenyl in the group may optionally be substituted with one, two or three R 3a optionally substituted with a group); (e1) optionally 1, 2 or 3 R 3a and may be substituted with -phenyl-LG 1 5- or 6-membered heteroaryl substituted with 1, 2, or 3 R 3aoptionally substituted with a group); (f1) optionally 1, 2 or 3 R 3a and may be substituted with a -(5- or 6-membered heteroaryl)-LG 1 5- or 6-membered heteroaryl substituted with -(5- or 6-membered heteroaryl)-LG 1 The 5- or 6-membered heteroaryl in 3a optionally substituted with a group); (h1) substituted with NH2 or OH and optionally further substituted with one or two R 3a C3-C6 cycloalkyl optionally substituted with a group; or (i1) C3-C6 cycloalkyl substituted with phenyl (wherein the phenyl is LG 1 and said phenyl is further optionally substituted with one, two or three R 3a groups, and the cycloalkyl may optionally be substituted with one or two R 3a (which may be substituted with a group) is) or a salt thereof, and / or a stereoisomer or mixture of stereoisomers; (However, the compound is: Methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate

[0020] [ka]

[0021] or its salts and / or its stereoisomers or mixtures of stereoisomers) is provided.

[0022] In a seventh aspect, a) Formula (A):

[0023] [ka]

[0024] The compound of R 2 contacting with -C(O)H; or b) Formula (B):

[0025] [ka]

[0026] The compound of R 2 -C(NH)NH2, where R 20 is Me or CD3); or c) Formula (C):

[0027] [ka]

[0028] The compound R 2 -H, wherein LG 1 is fluoro, chloro, bromo, iodo, triflate, mesylate, triazole, pyrazole, boronic acid, boronic ester, or allyltrifluoroborate; and optionally isolating the compound of formula (I). In one or more embodiments, the contacting is performed under basic conditions (in the presence of a base). [Brief explanation of the drawings]

[0029] [Figure 1] FIG. 1 shows the inhibition of Wnt transcriptional signaling pathway activity by a compound of formula (I) (Compound 1) compared to a compound of formula (I) and GO-HA (GO-HA + Compound 1) (Biological Example 3). [Figure 2] FIG. 2 shows the inhibition of Wnt transcriptional signaling pathway activity by a compound of formula (I) (compound 7) compared to a compound of formula (I) and GO-HA (GO-HA + compound 7) (Biological Example 3). [Figure 3]FIG. 3 shows the inhibition of Wnt transcriptional signaling pathway activity by a compound of formula (I) (compound 8) compared to a compound of formula (I) and GO-HA (GO-HA + compound 8) (Biological Example 3). [Figure 4] Figure 4 shows the results of a rabbit ear study (Biological Example 5), comparing eight injury sites (L1, L2, L3, L4, R5, R6, R7, and R8) of three specimens at day 0 (top) and day 23 (bottom). Specimens were treated with saline as a control and a compound of formula (I) (Compound 8 or Compound 18). Saline was applied by spray, Compound 8 (1 mg / mL) was applied by phospholipid spray, and Compound 18 (1 mg / mL) was applied by phospholipid spray (Biological Example 5). [Figure 5A] Figure 5A shows the rate of ear closure on days 1-21 (Study 1) compared to saline control and compounds of formula (I) (Compound 8 and Compound 18) (Biological Example 5). [Figure 5B] Figure 5B shows the rate of ear closure on days 14-39 (Study 1) compared to saline control and compounds of formula (I) (Compound 8 and Compound 18) (Biological Example 5). [Figure 6] Figure 6 shows the rate of ear closure on days 14-39 (Study 2) compared to saline control and compounds of formula (I) (Compound 8 and Compound 18). [Figure 7] Figure 7 (top) shows the slide and the location of the implant cut across the center of the healed wound for the purpose of processing the tissue after cartilage regeneration testing (studies in rabbit ears, see Biological Example 5). Figure 7 (bottom) shows a cross section of the sectioned tissue at the center of the wound. The resulting tissue sections, placed on the slide, are shown in the images in Figures 9-11. [Figure 8] 8 shows the mean distance (mm) between opposing cartilage endplates after 45 days in a cartilage regeneration assay (rabbit ear study). Measurements were taken from Safranin O-stained cross sections of healed ear punch wounds 45 days after treatment with saline (control) or a compound of formula (I) (Compound 8 or Compound 18). [Figure 9]Figure 9 shows a representative sample of a cross section taken on day 45 from a cartilage regeneration study (Biological Example 5) after treatment with saline (control). Analysis is shown 9-10 mm from the wound edge. The gray line represents the wound edge. The top image is at 0.3x magnification, the middle image is at 2.5x magnification, and the bottom image is at 10x magnification. The gray square in the middle image represents the image shown in the bottom image. The gray arrow in the bottom image indicates the area of ​​cartilage regeneration. [Figure 10] Figure 10 shows a representative sample of a cross section taken on day 45 from a cartilage regeneration study (Biological Example 5) after treatment with a compound of Formula (I) (Compound 8). Analysis is shown 9-10 mm from the wound edge. The gray line represents the wound edge. The top image is at 0.3x magnification, the middle image is at 2.5x magnification, and the bottom image is at 10x magnification. The gray square in the middle image represents the image shown in the bottom image. The gray arrow in the bottom image indicates the area of ​​cartilage regeneration. [Figure 11] Figure 11 shows a representative sample of a cross section taken on day 45 from a cartilage regeneration study (see Biological Example 5) after treatment with a compound of Formula (I) (Compound 18). Analysis is shown 9-10 mm from the wound edge. The gray line represents the wound edge; the top image is at 0.3x magnification, the middle image is at 2.5x magnification, and the bottom image is at 10x magnification. The gray square in the middle image represents the image shown in the bottom image. The gray arrow in the bottom image indicates the area of ​​cartilage regeneration. [Figure 12] FIG. 12 is a schematic diagram of the location of the wound site in a mammal (pig) in a full thickness excisional wound healing study. [Figure 13] Figure 13 (top) shows a suture formation with simple interrupted closure used in a full-thickness excision wound healing study (see Biological Example 6), showing a portion of a wound closed with a first throw 3401, a second throw 3403, and an interrupted suture 3405. Figure 13 (bottom) shows a cross-section of a portion of a wound closed with interrupted suture 3405. [Figure 14A] FIG. 14A shows the results of full-thickness open excision wound healing, epidermal ridge formation, in the compound of formula (I) and GO-HA (Compound 8 + GO-HA), saline control, and intact skin. [Figure 14B] FIG. 14B shows the results of epidermal protrusion formation (highest outliers excluded) of full-thickness open excision wound healing for compounds of formula (I) (Compound 7, Compound 8), and compounds of formula (I) and GO-HA (Compound 7 + GO-HA, Compound 8 + GO-HA), saline control, and GO-HA alone (p*>0.05; p**>0.01; and p***>0.001). [Figure 15] Figure 15 shows the results of a third-degree burn healing study (Biological Example 6) comparing a compound of Formula (I) (Compound 8) to a saline control. Wound area (cm2) was measured at intervals of 2 to 6 days. Results are shown from days 16 to 22. [Figure 16] Figure 16 shows the results of a third-degree burn healing study (Biological Example 6) comparing a compound of Formula (I) (Compound 7) to a saline control. Wound area (cm2) was measured at intervals of 2 to 6 days. Results are shown from days 16 to 22. [Figure 17] Figure 17 shows the results of a third-degree burn healing study (Biological Example 6) comparing a compound of formula (I) and GO-HA (Compound 8 + GO-HA) with a saline control. Wound area (cm2) was measured at intervals of 2 to 6 days. Results from days 16 to 22 are shown. [Figure 18] Figure 18 shows the results of a third-degree burn healing study (Biological Example 6) comparing a compound of formula (I) and GO-HA (Compound 7 + GO-HA) with a saline control. Wound area (cm2) was measured at intervals of 2 to 6 days. Results from days 16 to 22 are shown. [Figure 19] Figure 19 shows the results of a closed excision wound study (Biological Example 6): saline control, serum control (also known as serum formulation control), and GO-HA control, days 1 through 21. Sutures were removed on day 13. [Figure 20]Figure 20 shows the results of a closed excision wound study (Biological Example 6): Compounds of Formula (I) (Compound 1, Compound 8, and Compound 10), Days 1 to 21. Sutures were removed on Day 13. [Figure 21] Figure 21 shows the results of a closed excision wound study (Biological Example 6): Compounds of Formula (I) and GO-HA (Compound 1 + GO-HA, Compound 8 + GO-HA, and Compound 10 + GO-HA), days 1 to 21. Sutures were removed on day 13. [Figure 22] Figure 22 shows the results of a closed excision wound study (Biological Example 6): cross-sectional tissue samples stained with Trichrome Blue comparing wound sites treated with a saline control, a serum formulation control, a GO-HA control, a compound of Formula (I) (Compound 1, Compound 8, Compound 10), and a compound of Formula (I) and GO-HA (Compound 1 + GO-HA, Compound 8 + GO-HA, and Compound 10 + GO-HA). [Figure 23] Figure 23 shows the results of a closed excision wound study (see Biological Example 6): cross-sectioned tissue samples under polarized light microscopy comparing wound sites treated with saline control, serum formulation control, GO-HA control, compounds of formula (I) (Compound 1, Compound 8, Compound 10), and compounds of formula (I) and GO-HA (Compound 1 + GO-HA, Compound 8 + GO-HA, and Compound 10 + GO-HA). [Figure 24] Figure 24 shows the results of collagen infiltration (see Biological Example 6) of polarized images of closed excision wounds. The number of wounds showing collagen infiltration into the scar is shown for A - saline (also known as saline control); B - serum (also known as serum formulation control); C - GO-HA (also known as GO-HA control); D - Compound 1; E - Compound 1 + GO-HA; F - Compound 8; G - Compound 8 + GO-HA; H - Compound 10; and J - Compound 10 + GO-HA. [Figure 25]Figure 25 shows the results of a third-degree burn healing study (Biological Example 6) comparing compounds of formula (I) (Compound 7, Compound 7 + GO-HA, Compound 8, Compound 8 + GO-HA) with saline and GO-HA controls. Histology showed that the compounds promoted tissue regeneration (improved organized reticular collagen and epidermal ridge formation) and reduced scar formation compared to saline and GO-HA controls. DETAILED DESCRIPTION OF THE INVENTION

[0030] Detailed Description The present disclosure provides novel compounds, compositions, and methods of administration thereof for inducing wound healing, burn healing (including first, second, and third degree burns), or healing of lesions (including lesions caused by viruses selected from HPV and / or the Poxviridae family); treating inflammatory skin diseases, cartilage diseases, bone diseases, organ fibrosis, or cancer; inducing tissue regeneration (including, but not limited to, regeneration of damaged elastic cartilage); inducing bacteriostasis; inducing bacterial growth inhibition; maintaining bacteriostasis; inducing antifungal activity; inducing angiogenesis (in tissues in need); inducing reinnervation (in parts of the body that have lost nerves); inhibiting osteoclast differentiation; enhancing osteoblast differentiation; and / or inhibiting bone destruction associated with breast cancer.

[0031] The wound may include, but is not limited to, one or more selected from the group consisting of chronic wounds, acute wounds, and alkali-burned corneal wounds. The inflammatory skin disease may include, but is not limited to, one or more selected from the group consisting of acne, psoriasis, rosacea, and scleroderma. The cartilage disease may include, but is not limited to, one or more selected from the group consisting of osteoarthritis, rheumatoid arthritis, internal joint derangements, and degenerative cartilage disease. The bone disease may include, but is not limited to, bone diseases associated with bone formation disorders, such as osteoporosis.

[0032] Organ fibrosis may include, but is not limited to, one or more selected from the group consisting of pulmonary fibrosis, cardiac fibrosis, liver fibrosis, and kidney fibrosis.

[0033] The cancer may include, but is not limited to, melanoma, breast cancer, and / or prostate cancer.

[0034] Further provided herein are compounds, pharmaceutical compositions comprising the compounds, methods for making the compounds, and methods for using the compounds and compositions in treating wounds, particularly enhancing tissue regeneration after wound treatment. Also provided herein are methods for treating a condition associated with Wnt transcripts or Wnt signaling pathway activity in a mammal, comprising administering a therapeutically effective amount of the compound or composition to the mammal. In one or more embodiments, the mammal is a human.

[0035] definition When referring to the compounds provided herein, the following terms have the following meanings unless otherwise indicated. 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. If there are multiple definitions of a term herein, those in this section take precedence unless otherwise stated. Unless otherwise specified, when a term is defined as substituted, the group in the list of substituents is itself unsubstituted. For example, a substituted alkyl group can be substituted with, for example, a cycloalkyl group, and the cycloalkyl group is not further substituted unless otherwise specified.

[0036] Reference herein to "about" a value or parameter includes (and describes) a variation toward that value or parameter itself. For example, a statement referring to "about X" includes the statement "X." As used herein, and unless otherwise specified, the terms "about" and "approximately," when used in connection with a temperature, dose, amount, or weight percent of a component of a composition or dosage form, refer to a dose, amount, or weight percent that would be recognized by one of ordinary skill in the art as providing a pharmacological effect equivalent to that obtained from the specified dose, amount, or weight percent. Specifically, the terms "about" and "approximately," when used in this context, contemplate a dose, amount, or weight percent that is within 15%, 10%, 5%, 4%, 3%, 2%, 1%, or 0.5% of the specified dose, amount, or weight percent.

[0037] The terms "a" or "an," as used herein, mean one or more unless the context clearly dictates otherwise. For example, a "pharmaceutically acceptable carrier" includes one or more of the ingredients provided herein.

[0038] As used herein with respect to the GO-HA linker, "alkyl" refers to a straight-chain or branched-chain hydrocarbon. The alkyl may be straight-chain, branched-chain, cyclic, or a combination thereof and may contain, for example, 1 to 60 carbon atoms, and in some embodiments, 2 to 25 carbon atoms. Examples of alkyl groups include, but are not limited to, ethyl, propyl, isopropyl, cyclopropyl, butyl isomers (e.g., n-butyl, isobutyl, tert-butyl, etc.), cyclobutyl isomers (e.g., cyclobutyl, methylcyclopropyl, etc.), pentyl isomers, cyclopentyl isomers, hexyl isomers, cyclohexyl isomers, and the like.

[0039] The term "alkyl," as used herein with respect to compounds of Formula (I), unless otherwise specified, refers to a saturated, straight- or branched-chain monovalent hydrocarbon. In one or more embodiments, an alkyl group is a primary, secondary, or tertiary hydrocarbon. An alkyl may be straight- or branched-chain and may contain, for example, 1 to 8 carbon atoms. In one or more embodiments, an alkyl group has 1 to 6 carbon atoms, i.e., a C1-C6 alkyl. In one or more embodiments, an alkyl is a C 1-3 and alkyl. In one or more embodiments, the alkyl group is selected from the group consisting of methyl, ethyl, propyl, isopropyl, butyl, isobutyl, secbutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, isohexyl, 3-methylpentyl, 2,2-dimethylbutyl, and 2,3-dimethylbutyl. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl isomers (such as n-butyl, isobutyl, and tert-butyl), pentyl isomers, hexyl isomers, and the like.

[0040] As used herein with respect to the GO-HA linker, the term "straight chain alkyl" refers to a chain of carbon and hydrogen atoms (e.g., ethane, propane, butane, pentane, hexane, etc.).

[0041] As used herein with respect to the GO-HA linker, the term "branched alkyl" refers to a chain of carbon and hydrogen atoms with no double or triple bonds, and containing a fork, branch, and / or split in the chain. "Branching" refers to the divergence of the carbon chain, and "substitution" refers to the presence of a non-carbon / non-hydrogen atom in the moiety.

[0042] The term "alkylene," as used herein, unless otherwise specified, refers to a divalent alkyl group as defined herein for the GO-HA linker or, where applicable, for the compounds of formula (I).

[0043] As used herein with respect to the GO-HA linker, the term "cycloalkyl" refers to a fully saturated monocyclic or polycyclic hydrocarbon ring system. When composed of two or more rings, the rings may be joined together in a fused, bridged, or spiro-linked manner. Cycloalkyl groups may be unsubstituted, substituted, branched, and / or unbranched. Typical cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like. If substituted, the substituent(s) may be alkyl (but not substituted alkyl) or may be selected from those set forth above with respect to alkyl group substitution, unless otherwise specified. Unless otherwise specified (e.g., substituted cycloalkyl group, heterocyclyl, cycloalkoxy group, halocycloalkyl, cycloalkylamine, thiocycloalkyl, etc.), alkyl groups contain only carbon and hydrogen atoms. In some or all embodiments, cycloalkyl groups include cycloalkyls of 3 to 6 carbon atoms, i.e., C3 to C6. In some or all embodiments, cycloalkyls include cycloalkyls of 3, 4, 5, or 6 carbon atoms (C 3-6 ); 3, 4, or 5 (C 3-5 ); 3 or 4 (C 3-4 ; 3 (C3); 4 (C4); or 5 (C5) carbon atoms. In some or all embodiments, the cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. In some or all embodiments, the cycloalkyl group is cyclopropyl, cyclobutyl, or cyclopentyl. In one or more embodiments, the cycloalkyl group is cyclobutyl.

[0044] As used herein with respect to compounds of formula (I), the term "C3-C 10"-cycloalkyl" refers to a monovalent saturated monocyclic hydrocarbon or bicyclic (fused, bridged, or spirocyclic) ring. In some or all embodiments, the terms "fused cycloalkyl" and "spirocycloalkyl" are embodiments of cycloalkyl groups. In some or all embodiments, cycloalkyl groups include cycloalkyls of 3 to 6 carbon atoms, i.e., C3-C6. In some or all embodiments, cycloalkyls include cycloalkyls of 3, 4, or 5 carbon atoms (C 3-5 ); 3 or 4 (C 3-4 ; 3 (C3); 4 (C4); or 5 (C5) carbon atoms. In some or all embodiments, the cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. In some or all embodiments, the cycloalkyl group is cyclopropyl, cyclobutyl, or cyclopentyl. In some or all embodiments, the cycloalkyl group is cyclopropyl. In some or all embodiments, the cycloalkyl group is cyclobutyl. In some or all embodiments, the cycloalkyl group is cyclopentyl. In some or all embodiments, the cycloalkyl group is bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl, bicyclo[3.3.1]nonyl, bicyclo[3.3.2.]decyl, bicyclo[2.2.2]octyl, or adamantyl. In one or more embodiments, the cycloalkyl group is cyclobutyl.

[0045] The terms "alkoxy" and "alkyloxy," as used herein, and unless otherwise specified, refer to the group --OR', where R' is alkyl. In one or more embodiments, alkoxy is a group selected from the group consisting of C 1-6 Alkoxy. In one or more embodiments, alkoxy is C1-C3 alkoxy. Alkoxy and alkyloxy groups, in one or more embodiments, include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexyloxy, 1,2-dimethylbutoxy, and the like.

[0046] The term "alkoxyalkyl," as used herein with respect to compounds of Formula (I), and unless otherwise specified, refers to an alkyl group, as defined herein, substituted with one or two -OR' groups, where each R' is an independently selected alkyl, as defined herein. In some or all embodiments, the alkoxyalkyl is a C-C alkoxy-C-C alkyl.

[0047] The term "alkoxycarbonyl-NH-alkyl," as used herein with respect to compounds of Formula (I), and unless otherwise specified, refers to an alkyl group substituted with -NH-C(O)O(alkyl), where alkyl is as defined herein. In some or all embodiments, the alkoxycarbonyl-NH-alkyl is C-C alkoxycarbonyl-NH-C-C alkyl.

[0048] As used herein, "alkenyl," as used herein with respect to the GO-HA linker, means a straight or branched chain hydrocarbon having at least two carbon atoms containing at least one carbon-carbon double bond.

[0049] As used herein, "alkynyl," as used herein with respect to the GO-HA linker, means a straight or branched chain hydrocarbon having at least two carbon atoms containing at least one carbon-carbon triple bond.

[0050] As used herein, "amine" or "amino," as used herein for the GO-HA linker, is represented by the formula -NA1A2, where A1 and A2 are independently hydrogen or an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group, each of which groups is as described herein for the GO-HA linker. In one or more embodiments, amine (or amino) refers to any of NH2, NH(alkyl), NH(aryl), N(alkyl), N(alkyl)(aryl), and N(aryl).

[0051] The term "aryl," as used herein, and unless otherwise specified, refers to a monovalent C-C alkyl group containing at least one aromatic ring. 15 It refers to a carbocyclic ring system, where the aryl ring system is mono-, di-, or tricyclic. Aryl may be attached to the main structure through any of its rings, i.e., aromatic or non-aromatic. In some or all embodiments, aryl groups may be bridged (where chemically feasible) or unbridged, spirocyclic (where chemically feasible) or non-spirocyclic, and / or fused or non-fused polycyclic groups. In some or all embodiments, aryl is a C6-C 10 It is aryl. In some or all embodiments, the aryl is C6 aryl, i.e., phenyl. In some or all embodiments, the aryl is phenyl, naphthyl, indanyl, fluorenyl, 6,7,8,9-tetrahydro-5H-benzo[7]annulenyl, or tetrahydronaphthyl. When the aryl is substituted, it can be substituted on any ring, i.e., any aromatic or non-aromatic ring constituted by the aryl.

[0052] The term "haloalkyl," as used herein, and unless otherwise specified, refers to an alkyl group substituted with 1, 2, 3, 4, or 5 halo groups. In some or all embodiments, haloalkyl is halo-C 1-6In some or all embodiments, haloalkyl is —CF 3 , —CH 2 F, —CHF 2 , or —CH 2 CF 3 .

[0053] The terms "halogen" and "halo," as used herein, and unless otherwise specified, are synonymous and refer to chloro, bromo, fluoro, or iodo.

[0054] The term "heteroaryl," as used herein, and unless otherwise specified, refers to a heteroaryl group in which one or more (in some or all embodiments 1, 2, 3, or 4) ring atoms are O, S(O), 0-2

[0023] The term "heteroaryl" refers to a monocyclic or polycyclic aromatic ring system in which each heteroatom is independently selected from , NH, and N, the remaining ring atoms are carbon atoms, and the ring may be optionally substituted as described herein. A heteroaryl group is attached to the remainder of the molecule through any atom in the ring system, valence rules permitting. In some or all embodiments, each ring of a heteroaryl group can contain 1 or 2 O atoms, 1 or 2 S atoms, and / or 1 to 4 N atoms, or a combination thereof, provided that the total number of heteroatoms in each ring is 4 or less and each ring contains at least 1 carbon atom. In some or all embodiments, a heteroaryl has 5 to 20, 5 to 15, 5 to 6, or 5 to 10 ring atoms. When a heteroaryl is substituted, it may be substituted on any ring. In one or more embodiments, a heteroaryl is a 5- to 10-membered heteroaryl. In one or more embodiments, a heteroaryl is a 5- or 6-membered heteroaryl. In one or more embodiments, heteroaryl is a 6-membered heteroaryl. In one or more embodiments, heteroaryl is

[0055] [ka]

[0056] (In the formula,

[0057] [ka]

[0058] indicates the point of attachment of the heteroaryl to the rest of the molecule) is.

[0059] In some or all embodiments, monocyclic heteroaryl groups include, but are not limited to, furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl, and triazolyl. In some or all embodiments, bicyclic heteroaryl groups include, but are not limited to, benzofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzisothiazolyl, benzothienyl, benzotriazolyl, furopyridyl, thienopyridyl, imidazopyridinyl, imidazothiazolyl, indolizinyl, indolyl, indazolyl, isobenzofuranyl, isobenzothienyl, isoindolyl, isoquinolinyl, isothiazolyl, naphthyridinyl, oxazolopyridinyl, phthalazinyl, pteridinyl, purinyl, pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalinyl, or quinazolinyl. In some or all embodiments, tricyclic heteroaryl groups include, but are not limited to, acridinyl, benzoindolyl, carbazolyl, dibenzofuranyl, perimidinyl, phenanthrolinyl, phenanthridinyl, and phenazinyl. In some or all embodiments, heteroaryl is indolyl, furanyl, pyridinyl, pyrimidinyl, imidazolyl, or pyrazolyl; each of which is as defined throughout this specification and in some embodiments, C 1-6 Alkyl, hydroxy, halo, halo-C 1-6 Alkyl, C1-6 It may be optionally substituted with 1, 2, 3, or 4 groups, including group(s) independently selected from alkoxy, cyano, or phenyl.

[0060] The term "heterocyclic," as used herein, and unless otherwise specified, refers to a monovalent, monocyclic, non-aromatic ring system or a monovalent polycyclic ring system containing at least one non-aromatic ring; wherein one or more (in some or all embodiments, 1, 2, 3, or 4) of the monocyclic non-aromatic ring atoms are O, S(O), 0-2 and N, and the remaining ring atoms are carbon atoms; and any one or more (in some or all embodiments 1, 2, 3, or 4) of the ring atoms in the polycyclic ring system are O, S(O), 0-2、and N, and the remaining ring atoms are carbon. The term "heterocycle" does not include fully aromatic ring(s), i.e., it does not include imidazole, pyrimidine, pyridine, etc. In some or all embodiments, a heterocycle contains one or two heteroatom(s) independently selected from nitrogen and oxygen. In some or all embodiments, a heterocycle contains one or two heteroatom(s) that are oxygen. In some or all embodiments, a heterocycle contains one or two heteroatom(s) that are nitrogen (wherein the nitrogen is substituted as described in any aspect or embodiment described herein). In some or all embodiments, a heterocycle is polycyclic and contains one heteroatom in a non-aromatic ring, one heteroatom in an aromatic ring, two heteroatoms in an aromatic ring, or two heteroatoms, one in an aromatic ring and one in a non-aromatic ring. In some or all embodiments, a heterocyclic group has 3 to 20, 3 to 15, 3 to 10, 3 to 8, 4 to 7, or 5 to 6 ring atoms. In one or more embodiments, the heterocycle is a 4- to 10-membered heterocycle. In one or more embodiments, the heterocycle is a 5- to 10-membered heterocycle. In some or all embodiments, the heterocycle is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system. In some or all embodiments, the heterocyclic group can be a bridged or unbridged, spirocyclic or non-spirocyclic, and / or fused or non-fused polycyclic group. One or more nitrogen and sulfur atoms may be optionally oxidized, one or more nitrogen atoms may be optionally quaternized, and one or more carbon atoms may be optionally substituted.

[0061] [ka]

[0062] Some rings may be partially or fully saturated, or may be aromatic, provided the heterocycle is not fully aromatic. Monocyclic and polycyclic heterocycles may be attached to the main structure at any heteroatom or carbon atom that results in a stable compound. Polycyclic heterocycles may be attached to the main structure through any of their rings, including any aromatic or non-aromatic ring, regardless of whether the ring contains a heteroatom. In some or all embodiments, the heterocycle is a "heterocycloalkyl," which is 1) a saturated monovalent monocyclic group containing at least one ring heteroatom, as described herein, or 2) a saturated monovalent bicyclic or tricyclic group, as described herein, in which at least one ring contains at least one heteroatom. In some or all embodiments, the heterocycle is a 3- to 6-membered heterocycloalkyl. In some or all embodiments, the heterocycle is a 3- to 8-membered heterocycloalkyl. In some or all embodiments, the heterocycle is a 3- to 9-membered heterocycloalkyl. When heterocyclyl and heterocycloalkyl are substituted, they can be substituted on any ring, i.e., any aromatic or non-aromatic ring contained within the heterocyclyl and heterocycloalkyl. In some or all embodiments, such heterocycles include azepinyl, benzodioxanyl, benzodioxolyl, 3,4-dihydro-2H-benzo[b][1,4]oxazinyl, 3,4-dihydro-2H-benzo[b][1,4]dioxepinyl, 1,3-dihydroisobenzofuranyl, benzofuranonyl, benzopyranonyl, benzopyranyl, dihydrobenzofuranyl, benzotetrahydrothienyl, 2,2-dioxo-1,3-dihydrobenzo[c]thienyl, benzothiopyranyl, benzoxazinyl, β-carbolinyl, chromanyl, chromonyl, cinnolinyl, coumarinyl, decahydroquinolinyl, decahydroisoquinolinyl, dihydrobenzimidazolonyl (2-oxo-1,3-dihydro-2H-benzo[d]imidazol-1-yl), dihydrobenzisothiazinyl, dihydrobenzisoxazinyl, dihydrofuryl, dihydroisoindolyl, dihydropyranyl, dihydropyrazolyl, dihydropyrazinyl, dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl, dioxolanyl, 1,4-dithianyl, furanonyl, imidazolidinyl, 2,4-dioxo-imidazolidinyl, imidazolinyl, Indolinyl, 2-oxo-indolinyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothienyl, isochromanyl, isocoumarinyl, isoindolinyl, 1-oxo-isoindolinyl, 1,3-dioxo-isoindolinyl, isothiazolidinyl, isoxazolidinyl, 3-oxo-isoxazolidinyl, morpholinyl, 3,5-dioxo-morpholinyl, octahydroindolyl, octahydroisoindolyl, 1-oxo-octahydroisoindolyl aryl, 1,3-dioxo-hexahydroisoindolyl, oxo-oxadiazolyl (including but not limited to 5-oxo-1,2,4-oxadiazol-3-yl), oxazolidinonyl, oxazolidinyl, oxiranyl, piperazinyl, 2,6-dioxo-piperazinyl, piperidinyl, 2,6-dioxo-piperidinyl, 4-piperidonyl, pyrazolidinyl, pyrazolinyl, pyrrolidinyl, pyrrolinyl, 2-oxopyrrolidinyl, 2,5-dioxopyrrolidinyl In some or all embodiments, the heterocycle includes, but is not limited to, lysinyl, quinuclidinyl, tetrahydrofuryl, tetrahydroisoquinolinyl, tetrahydropyranyl, tetrahydrothienyl, thiamorpholinyl, thiomorpholinyl, 3,5-dioxothiomorpholinyl, thiazolidinyl, 2,4-dioxo-thiazolidinyl, tetrahydroquinolinyl, phenothiazinyl, phenoxazinyl, xanthenyl, 1,3,5-trithianyl, or 1,3-dihydro-imidazopyridin-2-onyl. In some or all embodiments, the heterocycle includes, but is not limited to, benzo-1,4-dioxanyl, benzodioxolyl, indolinyl, 2-oxo-indolinyl, pyrrolidinyl, piperidinyl, 2,3-dihydrobenzofuranyl, decahydroquinolinyl, dihydrocyclopentapyridyl, dihydropyranopyridyl, tetrahydronaphthyridyl, 2,In some embodiments, the heterocycloalkyl is 2-dioxo-3,4-dihydro-thiopyrano-pyridyl, dihydrofuropyridyl, dihydropyrrolopyridyl, 2,2-dioxo-1,3-dihydro-thieno-pyridyl, or tetrahydrocyclopropacyclopentapyridyl; each of which may be optionally substituted with 1, 2, 3, or 4 groups, as defined throughout this specification, including, in some or all embodiments, group(s) independently selected from halo, alkyl, and phenyl. In some embodiments, the heterocycloalkyl is pyrrolidinyl. In some embodiments, the heterocycloalkyl is an N-linked heterocycloalkyl.

[0063] The term "hydroxyalkyl," as used herein, unless otherwise specified, refers to an alkyl, as defined herein, substituted with one, two, or three hydroxy groups. In one or more embodiments, the hydroxy group is a primary, secondary, or tertiary alcohol. In one or more embodiments, the hydroxyalkyl group is a hydroxy group having from 1 to 10 carbons, i.e., C1 to C6. 10 In one or more embodiments, the hydroxyalkyl group contains one or two alcohol (hydroxy) groups, provided that they are not on the same carbon. In one or more embodiments, the hydroxyalkyl group contains a hydroxy C 1-6 In one or more embodiments, the hydroxyalkyl group is a hydroxyC 1-3 In one or more embodiments, the hydroxyalkyl group is selected from the group consisting of hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, hydroxypentyl, and hydroxyhexyl. In one or more embodiments, the hydroxyalkyl group is selected from the group consisting of C 1-6 In one or more embodiments, the hydroxyalkyl group is selected from the group consisting of hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 1-hydroxypropan-2-yl, and 2-hydroxypropan-2-yl.

[0064] The term "oxo," as used herein and unless otherwise specified, refers to a keto group (C=O). An oxo group that is a substituent of a non-aromatic carbon results in the conversion of -CH2- to -C=O. An oxo group that is a substituent of an aromatic carbon results in the conversion of -CH- to -C=O. When a substituent is oxo, two hydrogens on the atom are replaced. When an oxo group replaces an aromatic moiety, the corresponding partially unsaturated ring replaces the aromatic ring. For example, a pyridyl group substituted with an oxo group is a pyridone. Those skilled in the art will understand that in some embodiments, such groups, e.g., pyridone and 2,4(1H,3H)-dioxo-pyrimidinyl, can exist in their tautomeric forms, e.g., hydroxypyridine and 2,4-dihydroxypyrimidinyl, respectively.

[0065] As used herein, "regeneration" means the renewal or growth of destroyed or devitalized tissue from remaining tissue. It is the body's attempt to repair itself, and in the context of wounds, refers to the migration, differentiation, replication, or transformation of cells from progenitor cells into tissue-appropriate cell types (which may include sebocytes, hair follicles, nerve cells, and chondrocytes).

[0066] As used herein, "wound" refers to scrapes, cuts, abrasions, tissue or skin damage caused by surgical procedures (e.g., caused by minimally invasive surgery, laparoscopic surgery, robotic surgery, open biopsy, general surgery, and cosmetic surgery), stripped skin, burns, ulcers (e.g., diabetic ulcers, ulcers from vascular insufficiency, pressure sores, and burns), or other skin problems (e.g., allergies). Wounds can range from superficial (e.g., those affecting only the epidermis) to more traumatic (e.g., lesions affecting deeper skin or tissue layers beneath the epidermis). Wounds can be of any length or shape; for example, in some embodiments, wounds are straight, jagged, or curved.

[0067] In some embodiments, the term "pharmaceutically acceptable carrier" includes any and all, and / or one or more, solvents, cosolvents, complexing agents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, excipients, diluents, disintegrants, lubricants, adjuvants, etc. that are not biologically or otherwise undesirable. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the composition. In addition, various adjuvants commonly used in the art may be included. These and other such compounds are described in the literature, e.g., The Merck Index (Merck & Company, Rahway, NJ), and considerations for including various ingredients in pharmaceutical compositions are described (e.g., Gilman et al. (eds.), 2010, Goodman and Gilman's: The Pharmacological Basis of Therapeutics, 12th Edition, The McGraw-Hill Companies).

[0068] In some embodiments, the term "pharmaceutically acceptable salts" refers to salts that retain the biological effectiveness and properties of the compounds provided herein and are not biologically or otherwise undesirable. In many cases, the compounds provided herein may form acid and / or base salts due to the presence of amino and / or carboxyl groups or groups similar thereto. Inorganic and organic acids may be used to form pharmaceutically acceptable acid addition salts. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Inorganic and organic bases may be used to form pharmaceutically acceptable base addition salts. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, etc.; particularly preferred are ammonium, potassium, sodium, calcium, and magnesium salts. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, etc., specifically, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. Many of these salts are known in the art, as described in WO 87 / 05297.

[0069] In some embodiments, the term "pharmaceutically acceptable salt," as used herein, and unless otherwise specified, refers to any salt of a compound provided herein that retains its biological properties and is not toxic or otherwise desirable for pharmaceutical use. Such salts can be derived from a variety of organic and inorganic counterions well known in the art.Such salts include (1) hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, sulfamic acid, acetic acid, trifluoroacetic acid, trichloroacetic acid, propionic acid, hexanoic acid, cyclopentylpropionic acid, glycolic acid, glutaric acid, pyruvic acid, lactic acid, malonic acid, succinic acid, sorbic acid, ascorbic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, picric acid, cinnamic acid, mandelic acid, phthalic acid, lauric acid, methanesulfonic acid, ethanesulfonic acid, 1,2-dimethyl-2,3 ... -Ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphoric acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfuric acid, gluconic acid, benzoic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, cyclohexylsulfamic acid, chitosan, and (2) base addition salts formed when an acidic proton present in the parent compound is replaced by (a) a metal ion, e.g., an alkali metal ion, an alkaline earth metal ion, an aluminum ion, or an alkali metal or alkaline earth metal hydroxide, e.g., sodium, potassium, calcium, magnesium, aluminum, lithium, zinc, and barium hydroxide, ammonia, or (b) an organic base, e.g., an aliphatic, alicyclic, or aromatic organic amine, e.g., ammonia, methylamine, dimethylamine, diethylamine, picoline, ethanolamine, diethanolamine, triethanolamine, ethylenediamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylene-diamine, chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine, N-methylglucamine tris(hydroxymethyl)aminomethane, piperazine, tetramethylammonium hydroxide, and the like.

[0070] Pharmaceutically acceptable salts in some embodiments further include, in some or all embodiments, and without limitation, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium salts, etc. When the compound contains a basic functional group, salts of non-toxic organic or inorganic acids, such as hydrohalides, e.g., hydrochloride and hydrobromide, sulfate, phosphate, sulfamate, nitrate, acetate, trifluoroacetate, trichloroacetate, propionate, hexanoate, cyclopentylpropionate, glycolate, glutarate, pyruvate, lactate, malonate, succinate, sorbate, ascorbate, malate, maleate, fumarate, tartrate, citrate, benzoate, 3-(4-hydroxybenzoyl)benzoate, picrate, cinnamate, etc. , mandelate, phthalate, laurate, methanesulfonate (mesylate), ethanesulfonate, 1,2-ethanedisulfonate, 2-hydroxyethanesulfonate, benzenesulfonate (besylate), 4-chlorobenzenesulfonate, 2-naphthalenesulfonate, 4-toluenesulfonate, camphorate, camphorsulfonate, methylbicyclo[2.2.2]-oct-2-ene-1-carboxylate, glucoheptonate, 3-phenylpropionate, trimethylacetate, tert-butylacetate, lauryl sulfate, gluconate, benzoate, glutamate, hydroxynaphthoate, salicylate, stearate, cyclohexylsulfamate, quinate, muconate, etc.

[0071] In some embodiments, a "therapeutically effective amount" or "pharmaceutically effective amount" of a compound provided herein is an amount sufficient to achieve a desired effect, which may vary depending on the nature and severity of the disease state and the potency of the compound. A "therapeutically effective amount" is also intended to include one or more compositions of the present disclosure to result in increased regeneration of injured tissue. The combination of compounds is preferably a synergistic combination. Synergy occurs when the effect of compounds administered in combination is greater than the additive effect of the compounds administered alone as single agents, as described in the art (e.g., Chou, 2010, Canc. Res. 70(2):440-446). Generally, synergistic effects are most clearly demonstrated at suboptimal concentrations of the compounds. This amount may further depend on the patient's height, weight, sex, age, and medical history.

[0072] The term "mammal" specifically includes humans, cows, horses, dogs, and cats, but also includes many other mammalian species such as pigs, rats, mice, primates (e.g., monkeys such as cynomolgous monkeys, chimpanzees, and humans). In some embodiments, the mammal is a human.

[0073] The term "subject" refers to a mammal, and a cell or biological sample as provided herein.

[0074] The term "substantially free of" stereoisomers, with respect to a composition, refers to a composition that contains at least 85 or 90%, and in some or all embodiments 95%, 98%, 99%, or 100% by weight of a specified stereoisomer of a compound in the composition. In some or all embodiments, in the methods and compounds provided herein, the compound is substantially free of stereoisomers.

[0075] Similarly, with respect to a composition, the term "isolated" refers to a composition that contains at least 85, 90%, 95%, 98%, 99% to 100% by weight of the specified compound, with the remainder containing other species or stereoisomers.

[0076] The term "isotopic composition," as used herein, and unless otherwise specified, refers to the amount of each isotope present for a given atom, and "natural isotopic composition" refers to the naturally occurring isotopic composition or abundance for a given atom. Atoms containing natural isotopic compositions are also referred to herein as "non-enriched" atoms. Unless otherwise specified, atoms of compounds referred to herein are intended to represent all stable isotopes of that atom. For example, unless otherwise specified, if a position is specifically designated as "H" or "hydrogen," it is understood that the position has hydrogen at its natural isotopic composition.

[0077] The term "isotopic enrichment," as used herein and unless otherwise specified, refers to the proportion of a given atom in a molecule that incorporates a specific amount of isotope, instead of the natural isotopic abundance of that atom. In some or all embodiments, a deuterium enrichment of 1% at a given position means that 1% of the molecules in a given sample contain deuterium at that specified position. Since the distribution of naturally occurring deuterium is approximately 0.0156%, the deuterium enrichment at any position of a compound synthesized using non-enriched starting materials is approximately 0.0156%. The isotopic enrichment of the compounds provided herein can be determined using conventional analytical methods known to those skilled in the art, including mass spectrometry and nuclear magnetic resonance spectroscopy.

[0078] The term "isotopically enriched," as used herein, and unless otherwise specified, refers to an atom having an isotopic composition other than the natural isotopic composition of that atom. "Isotopically enriched" can also refer to a compound containing at least one atom having an isotopic composition other than the natural isotopic composition of that atom.

[0079] As used herein, and unless otherwise specified, the term "IC 50 " refers to the amount, concentration, or dosage of a particular test compound that achieves 50% inhibition of a maximal response in an assay that measures such response.

[0080] In some embodiments, the terms "therapeutic agent" and "therapeutic agents" refer to any drug or drugs that can be used in the treatment or prevention of a disorder, or one or more symptoms thereof. In some or all embodiments, the term "therapeutic agent" includes the compounds provided herein. In some or all embodiments, a therapeutic agent is an agent that is known to be useful, has been used, or is currently being used, in the treatment or prevention of a disorder, or one or more symptoms thereof.

[0081] In some embodiments, "treating" or "treatment" of any condition or disorder refers, in some or all embodiments, to improving the condition or disorder present in a subject. In other embodiments, "treating" or "treatment" includes improving at least one physical parameter, which may be indiscernible by the subject. In yet other embodiments, "treating" or "treatment" includes modulating the condition or disorder, either physically (e.g., stabilization of discernible symptoms) or physiologically (e.g., stabilization of physical parameters), or both. In yet other embodiments, "treating" or "treatment" includes delaying the onset of the condition or disorder. In yet other embodiments, "treating" or "treatment" includes alleviating or eliminating the condition, or one or more symptoms of the condition, or slowing the progression of the condition, or one or more symptoms of the condition, or alleviating the signs of the condition, or one or more symptoms of the condition.

[0082] compound Provided herein are compounds that can induce improved wound healing and tissue regeneration. Provided herein are compounds that can treat wounds and / or conditions associated with Wnt transcripts or Wnt signaling pathway activity, and in particular, can enhance tissue regeneration after wound treatment.

[0083] Further provided herein are compounds that can modulate the activity of the Wnt signaling pathway or Wnt transcripts, which can be formulated as described herein and used to treat conditions associated with Wnt transcripts or Wnt signaling pathway activity. In one or more embodiments, the condition associated with Wnt transcripts or Wnt signaling pathway activity is a chronic wound, an acute wound, an alkali-burned corneal wound, a burn, a lesion (including lesions caused by a virus selected from HPV and / or the Poxviridae family of viruses), an inflammatory skin disease (including acne, psoriasis, rosacea, and scleroderma), a cartilage disease (including osteoarthritis, rheumatoid arthritis, internal joint derangements, and degenerative cartilage diseases), a bone disease (including osteoporosis), an organ fibrosis (including pulmonary fibrosis, cardiac fibrosis, liver fibrosis, and kidney fibrosis), a cancer (including melanoma, breast cancer, and prostate cancer), a denervated body part in need of reinnervation, a tissue in need of regeneration (including damaged elastic cartilage), bacterial growth in need of inhibition, fungal growth in need of inhibition, a tissue in need of angiogenesis, osteoclast differentiation in need of inhibition, impaired osteoblast differentiation (where inhibition of osteoblast differentiation is required), and / or bone destruction associated with breast cancer.

[0084] The present disclosure arises from the novel and unexpected discovery of significant Wnt inhibition by compounds of Formula (I) (or any embodiment thereof, including in some embodiments Compound 1, Compound 7, or Compound 8), and with several improved properties compared to prior art Wnt inhibitors, such as XAV939. Some improved properties may include improved wound healing and regeneration of wounded tissue in mammals, both in the amount of regeneration and the quality of tissue regeneration and regrowth.

[0085] The aspects and embodiments described herein include the compounds mentioned and their pharmaceutically acceptable salts and / or isomers thereof. For example, the aspects and embodiments described herein include a single stereoisomer of a mixture of stereoisomers thereof, and / or a pharmaceutically acceptable salt thereof.

[0086] All stereoisomers of the compounds, including diastereomers and enantiomers, are included herein, where chemically possible. Also included are mixtures of possible stereoisomers in any ratio, including, but not limited to, racemic mixtures. Unless stereochemistry is explicitly stated in a structure at a particular atom, the structure is intended to encompass all possible stereoisomers of the depicted compound. When stereochemistry is explicitly stated for one or more portions of a molecule but not for another portion or portions of a molecule, the structure is intended to encompass all possible stereoisomers of the portion or portions where stereochemistry is not explicitly stated. It is clear that a structure refers to a specific stereochemistry at a particular atom.

[0087] 40. A composition for inhibiting Wnt transcription or Wnt signaling pathway activity, comprising: a matrix component comprising a conjugate of graphene oxide (GO) and hyaluronic acid (HA) (GO-HA), wherein GO and HA are covalently bonded via a linker; polyethylene glycol (PEG), wherein the PEG is optional; a thickening agent, wherein the thickening agent is optional; the compound of any one of claims 1-39; and water, wherein the compound optionally constitutes from about 0.001 wt% to about 5 wt% of the total composition.

[0088] In one or more embodiments, the compounds of formula (I) are potent inhibitors of the Wnt pathway.

[0089] Embodiment 1: In one or more embodiments of Formula (I), the compound is a potent inhibitor of the Wnt pathway (Compound 1), whose chemical name is (4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)boronic acid. The structure of Compound 1 is:

[0090] [ka]

[0091] In one or more embodiments, the compound of formula (I) is Compound 1, or any prodrug, pharmaceutically acceptable salt, metabolite, polymorph, solvate, hydrate, stereoisomer, or tautomer thereof.

[0092] Embodiment 2: In one or more embodiments of Formula (I), the compound is a potent inhibitor of the Wnt pathway having the chemical name 2-(4-(6-bromopyridin-3-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (Compound 7). The structure of Compound 7 is:

[0093] [ka]

[0094] In one or more embodiments, the compound of formula (I) is compound 7, or any prodrug, pharmaceutically acceptable salt, metabolite, polymorph, solvate, hydrate, stereoisomer, or tautomer thereof.

[0095] Embodiment 3: In one or more embodiments of Formula (I), the compound is a potent inhibitor of the Wnt pathway having the chemical name 2-(4-(2-(2-hydroxyethoxy)propan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (Compound 8). The structure of Compound 8 is:

[0096] [ka]

[0097] In one or more embodiments, the compound of formula (I) is compound 8, or any prodrug, pharmaceutically acceptable salt, metabolite, polymorph, solvate, hydrate, stereoisomer, or tautomer thereof.

[0098] Embodiment A In one or more embodiments, R 1 and R 1a is independently selected from hydrogen and deuterium.

[0099] Embodiment B In one or more embodiments, including any of the above embodiments, Ring A is

[0100] [ka]

[0101] (In the formula,

[0102] [ka]

[0103] indicates the bond to the remainder of the compound of formula (I) Compounds of formula (I) are provided, wherein:

[0104] The embodiment where R2 is (a) In one or more embodiments, including embodiments A and B, R 2 But R 3 and optionally 1, 2 or 3 R 3a In another aspect, a compound of formula (I) is provided, wherein R is phenyl, optionally substituted with a group.

[0105] In one or more embodiments, including embodiments A and B, R 3 is attached to the para position of the phenyl ring; or R 3 and one R 3a are located on adjacent carbon atoms, they form a ring (a-1) together with the carbon atoms to which they are attached, and The phenyl moiety may optionally be joined to the remaining R 3a optionally substituted with a group, Compounds of formula (I) are provided:

[0106] In one or more embodiments, including embodiments A and B, R 2 When (a) is true, R 3 is not halo or haloalkyl.

[0107] In one or more embodiments, including embodiments A and B, R 2 When (a) is true, R 3 is B(O)2 or -(C0-C6 alkylene)-OR 4 Compounds of formula (I) are provided, wherein:

[0108] In one or more embodiments, including embodiments A and B, R 2 but,

[0109] [ka]

[0110] In one or more embodiments, R 3 is not halo or haloalkyl. In one or more embodiments, R 3 is B(O)2 or -(C0-C6 alkylene)-OR 4 is.

[0111] In one or more embodiments, including embodiments A and B, R 2 but,

[0112] [ka]

[0113] In one or more embodiments, R 2 teeth,

[0114] [ka]

[0115] isn't it.

[0116] In one or more embodiments, including embodiments A and B, R 2 but,

[0117] [ka]

[0118] In one or more embodiments, R 2 teeth

[0119] [ka]

[0120] isn't it.

[0121] The embodiment where R2 is (b) In one or more embodiments, including embodiments A and B, R 2 is phenyl substituted with a 5- or 6-membered heteroaryl, wherein said 5- or 6-membered heteroaryl is R 3 and optionally 1, 2 or 3 R 3a groups, and the phenyl may further optionally be substituted with one, two or three R 3a The present invention provides a compound of formula (I), which may be substituted with a group:

[0122] In one or more embodiments, including embodiments A and B, R 2 is phenyl substituted at the para position with a 5- or 6-membered heteroaryl, wherein the 5- or 6-membered heteroaryl is R 3 and optionally 1, 2 or 3 R 3agroups, and the phenyl may further optionally be substituted with one, two or three R 3a and when said 5- or 6-membered heteroaryl is a 6-membered heteroaryl, R 3 is substituted at the para position of said 6-membered heteroaryl.

[0123] In one or more embodiments, including embodiments A and B, R 2 but,

[0124] [ka]

[0125] There is provided a compound of formula (I), wherein the compound is selected from the group consisting of:

[0126] In one or more embodiments, including embodiments A and B, R 2 but,

[0127] [ka]

[0128] There is provided a compound of formula (I), wherein the compound is selected from the group consisting of:

[0129] R 2 is (c). In one or more embodiments, including embodiments A and B, R 2 optionally 1, 2 or 3 R 3a and may be substituted with -phenyl-R 3 (wherein the -phenyl-R 3 The phenyl may have one, two or three R 3a The present invention provides a compound of formula (I), optionally substituted with a group:

[0130] In one or more embodiments, including embodiments A and B, R 2 optionally 1, 2 or 3 R 3a group, and further at the para position, 3 (wherein the -phenyl-R 3 The phenyl may optionally be substituted with one, two or three R 3a and R 3 is the phenyl-R 3 wherein the phenyl is in the para position of the phenyl.

[0131] In one or more embodiments, including embodiments A and B, R 2 but,

[0132] [ka]

[0133] There is provided a compound of formula (I), wherein the compound is selected from the group consisting of:

[0134] In one or more embodiments, including embodiments A and B, R 2 but,

[0135] [ka]

[0136] There is provided a compound of formula (I), wherein the compound is selected from the group consisting of:

[0137] R 2 is (d). In one or more embodiments, including embodiments A and B, R 2 But R 3 and optionally 1, 2 or 3 R 3a and 5-6 membered heteroaryl optionally substituted with a group (wherein R 3is in the para position of said 6-membered heteroaryl.

[0138] In one or more embodiments, including embodiments A and B, R 2 When (d) is true, R 3 is not halo or haloalkyl.

[0139] In one or more embodiments, including embodiments A and B, R 2 When (d) is true, R 3 is B(O)2 or -(C0-C6 alkylene)-OR 4 Compounds of formula (I) are provided, wherein:

[0140] In one or more embodiments, including embodiments A and B, R 2 When (d) is true, R 3 is B(O)2 or -(C1-C6 alkylene)-OR 4 Compounds of formula (I) are provided, wherein:

[0141] The embodiment where R2 is (e) In one or more embodiments, including embodiments A and B, R 2 optionally 1, 2 or 3 R 3a and may be substituted with -phenyl-R 3 wherein said phenyl is further optionally substituted with one, two or three R 3a optionally substituted with the -phenyl-R 3 is in the para position of said 6-membered heteroaryl; and optionally R 3 is in the para position of the phenyl.

[0142] The embodiment where R2 is (f) In one or more embodiments, including embodiments A and B, R 2is a 5- or 6-membered heteroaryl, said 5- or 6-membered heteroaryl optionally containing 1, 2, or 3 R 3a and may be substituted with a -(5- or 6-membered heteroaryl)-R 3 wherein the -(5- or 6-membered heteroaryl)-R 3 The 5- or 6-membered heteroaryl in 3a group; optionally, the -(6-membered heteroaryl)-R 3 is in the para position of the first 6-membered heteroaryl; and optionally R 3 is in the para position of the 6-membered heteroaryl to which it is attached.

[0143] In one or more embodiments, including embodiments A and B, R 2 but,

[0144] [ka]

[0145] Compounds of formula (I) are provided, wherein:

[0146] The embodiment where R2 is (g) In one or more embodiments, including embodiments A and B, R 2 But R 3 and optionally one or two R 3a The compound of formula (I) is provided wherein the C3-C6 cycloalkyl optionally substituted with a group.

[0147] In one or more embodiments, including embodiments A and B, R 2 but,

[0148] [ka]

[0149] (where R 3 is a 5-10 membered heterocycle optionally substituted with cyano.

[0150] The embodiment where R2 is (h) In one or more embodiments, including embodiments A and B, R 2 is a C3-C6 cycloalkyl substituted with a 5- or 6-membered heteroaryl, wherein the 5- or 6-membered heteroaryl is R 3 and optionally 1, 2 or 3 R 3a groups, and the cycloalkyl may optionally be substituted with one or two R 3a The present invention provides a compound of formula (I), which may be substituted with a group:

[0151] R 2 is (i) In one or more embodiments, including embodiments A and B, R 2 is a C3-C6 cycloalkyl substituted with phenyl, wherein said phenyl is R 3 and optionally 1, 2 or 3 R 3a groups, and the cycloalkyl may optionally be substituted with one or two R 3a The present invention provides a compound of formula (I), which may be substituted with a group:

[0152] The embodiment where R2 is (j) In one or more embodiments, including embodiments A and B, R 2 is a 3- to 8-membered heterocycloalkyl substituted with phenyl or a 5- or 6-membered heteroaryl, wherein the phenyl and the 5- to 6-membered heteroaryl are R 3 and optionally 1, 2 or 3 R 3a The present invention provides a compound of formula (I), which may be substituted with a group:

[0153] In one or more embodiments, including embodiments A and B, R 2 but

[0154] [ka]

[0155] There is provided a compound of formula (I), wherein the compound is selected from the group consisting of:

[0156] The embodiment where R2 is (k) In one or more embodiments, including embodiments A and B, R 2 But -CH=CH-R 5 (where R 5 is phenyl or 5- or 6-membered heteroaryl, and said phenyl and said 5- or 6-membered heteroaryl are R 3 and optionally 1, 2 or 3 R 3a The present invention provides a compound of formula (I), which may be substituted with a group:

[0157] In one or more embodiments, including embodiments A and B, R 2 but,

[0158] [ka]

[0159] There is provided a compound of formula (I), wherein the compound is selected from the group consisting of:

[0160] Additional Embodiments Including embodiments A and B, and R 2 In one or more embodiments, including any of the embodiments provided above when is (a) through (k), R 3 is cyano, -B(OH)2, or -(C0-C6 alkylene)-OR 4 In one or more embodiments, compounds of formula (I) are provided wherein R 4is hydroxy-C1-C6 alkyl (in some embodiments, hydroxyethyl). 4 In one or more embodiments, compounds of formula (I) are provided wherein R is C-C alkoxy-C-C alkyl (in some embodiments, C-C alkoxy-C alkyl). 4 is C1-C6 alkoxycarbonyl-NH-C1-C6 alkyl (in some embodiments, C1-C6 alkoxycarbonyl-NH-C2 alkyl).

[0161] Including embodiments A and B, and R 2 In one or more embodiments, including any of the embodiments provided above when is (a) through (k), R 3 is cyano.

[0162] Including embodiments A and B, and R 2 In one or more embodiments, including any of the embodiments provided above when is (a) through (k), R 3 is —B(OH) 2 .

[0163] Including embodiments A and B, and R 2 In one or more embodiments, including any of the embodiments provided above when is (a) through (k), R 3 -(C0-C6 alkylene)-OR 4 In one or more embodiments, compounds of formula (I) are provided wherein R 4 is hydroxy-C1-C6 alkyl (in some embodiments, hydroxyethyl). 4 In one or more embodiments, compounds of formula (I) are provided wherein R is C-C alkoxy-C-C alkyl (in some embodiments, C-C alkoxy-C alkyl).4 is C1-C6 alkoxycarbonyl-NH-C1-C6 alkyl (in some embodiments, C1-C6 alkoxycarbonyl-NH-C2 alkyl).

[0164] Including embodiments A and B, and R 2 In one or more embodiments, including any of the embodiments provided above when is (a) through (k), R 3 However, -(C 1-6 alkylene)-OR 4 In one or more embodiments, compounds of formula (I) are provided wherein R 4 is hydroxy-C1-C6 alkyl (in some embodiments, hydroxyethyl). 4 In one or more embodiments, compounds of formula (I) are provided wherein R is C-C alkoxy-C-C alkyl (in some embodiments, C-C alkoxy-C alkyl). 4 is C1-C6 alkoxycarbonyl-NH-C1-C6 alkyl (in some embodiments, C1-C6 alkoxycarbonyl-NH-C2 alkyl).

[0165] Including embodiments A and B, and R 2 In one or more embodiments, including any of the embodiments provided above when R is (b), (c), (e), or (f), 3 is halo, cyano, -B(OH)2, or -(C0-C6 alkylene)-OR 4 In one or more embodiments, compounds of formula (I) are provided wherein R 4 is hydroxy-C1-C6 alkyl (in some embodiments, hydroxyethyl). 4In one or more embodiments, compounds of formula (I) are provided wherein R is C-C alkoxy-C-C alkyl (in some embodiments, C-C alkoxy-C alkyl). 4 is C1-C6 alkoxycarbonyl-NH-C1-C6 alkyl (in some embodiments, C1-C6 alkoxycarbonyl-NH-C2 alkyl).

[0166] In one or more embodiments, including embodiments A and B, and including any embodiment provided above, R 2 is (b), (c), (e) or (f), then R 3 is halo, cyano, -B(OH)2, or -(C0-C6 alkylene)-OR 4 or a compound of formula (I) is provided, wherein R 2 is (b), (c), (e) or (f), then R 3 is cyano, -B(OH)2, or -(C0-C6 alkylene)-OR 4 Compounds of formula (I) are provided, wherein: In one or more embodiments, R 4 is hydroxy-C1-C6 alkyl (in some embodiments, hydroxyethyl). 4 In one or more embodiments, compounds of formula (I) are provided wherein R is C-C alkoxy-C-C alkyl (in some embodiments, C-C alkoxy-C alkyl). 4 is C1-C6 alkoxycarbonyl-NH-C1-C6 alkyl (in some embodiments, C1-C6 alkoxycarbonyl-NH-C2 alkyl).

[0167] In one or more embodiments, a compound of Formula (I) is provided that is selected from the group consisting of compounds 1-44 provided in Table 1.

[0168] In one or more embodiments, a pharmaceutical composition is provided comprising a compound of Formula (I); and a pharmaceutically acceptable carrier. In one or more embodiments, a pharmaceutical composition is provided comprising a compound of Formula (I), or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

[0169] In one or more embodiments, the pharmaceutical compositions herein comprise a matrix component comprising a conjugate of graphene oxide (GO) and hyaluronic acid (HA) (GO-HA), where GO and HA are covalently bonded via a linker; polyethylene glycol (PEG), where said PEG is optional; a thickening agent, where said thickening agent is optional; and water, optionally wherein said compounds optionally comprise from about 0.001 wt % to about 5 wt % of the total composition.

[0170] In one or more embodiments, provided is a method of inhibiting Wnt transcription or Wnt signaling pathway activity in a subject, comprising contacting the subject with an effective amount of a compound of Formula (I), or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof.

[0171] In one or more embodiments, provided is a method of treating a disease, disorder, or condition associated with Wnt transcription or Wnt signaling pathway activity in a mammal, comprising administering to a subject in need thereof a compound of Formula (I), or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof; or a pharmaceutical composition according to one or more embodiments herein.

[0172] In one or more embodiments, provided is a method of stimulating tissue regeneration at a wound in a subject in need thereof, comprising contacting the wound with an effective amount of a compound of formula (I) (or a single or mixture of stereoisomers thereof, a single or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition according to one or more embodiments herein.

[0173] In one or more embodiments, the disease, disorder, or condition (treated by one or more embodiments of the methods herein) is selected from chronic wounds, acute wounds, alkali-burned corneal wounds, burns, lesions (including lesions caused by viruses selected from HPV and / or Poxviridae), inflammatory skin diseases (including acne, psoriasis, rosacea, and scleroderma), cartilage diseases (including osteoarthritis, rheumatoid arthritis, internal joint derangements, and degenerative cartilage diseases), bone diseases (including osteoporosis), organ fibrosis (including pulmonary fibrosis, cardiac fibrosis, liver fibrosis, and kidney fibrosis), cancer (including melanoma, breast cancer, and prostate cancer), denervated body parts in need of reinnervation, tissues in need of regeneration (including damaged elastic cartilage), bacterial growth in need of inhibition, fungal growth in need of inhibition, tissues in need of angiogenesis, osteoclast differentiation in need of inhibition, impaired osteoblast differentiation (where inhibition of osteoblast differentiation is necessary), and / or bone destruction associated with breast cancer.

[0174] In one or more embodiments, provided are methods of inducing bacteriostasis associated with Wnt transcripts or Wnt signaling pathway activity, comprising administering to a mammal in need thereof XAV939, or a tautomer and / or a pharmaceutically acceptable salt thereof, optionally as a pharmaceutical composition thereof; administering a compound of Formula (I), or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof; or administering a pharmaceutical composition according to one or more embodiments herein.

[0175] In one or more embodiments, the compound has the formula:

[0176] [ka]

[0177] (In the formula, LG 1 is a leaving group such as fluoro, chloro, bromo, iodo, triflate, mesylate, triazole, pyrazole, boronic acid, boronic ester, or allyltrifluoroborate; R 1 is hydrogen, deuterium, C1-C3 alkyl, —OH, —O—C1-C3 alkyl, —CH2OH, or —B(OH)2; R 1a is hydrogen, deuterium, or C1-C3 alkyl; R 20 is alkyl, preferably methyl or ethyl, or CD; R 2 (b1) phenyl substituted with 5- or 6-membered heteroaryl, wherein the 5- or 6-membered heteroaryl is LG 1 and optionally 1, 2 or 3 R 3a groups, and the phenyl may further optionally be substituted with one, two or three R 3a (c1) optionally substituted with 1, 2 or 3 R 3a and may be substituted with -phenyl-LG 1 phenyl substituted with -phenyl-LG 1 The phenyl in the group may optionally be substituted with one, two or three R 3a (e1) optionally substituted with 1, 2 or 3 R 3a and may be substituted with -phenyl-LG 1 5- or 6-membered heteroaryl substituted with 1, 2, or 3 R 3a (f1) optionally substituted with 1, 2 or 3 R 3a and may be substituted with a -(5- or 6-membered heteroaryl)-LG 1 5- or 6-membered heteroaryl substituted with -(5- or 6-membered heteroaryl)-LG 1The 5- or 6-membered heteroaryl in 3a (h1) substituted with NH or OH and optionally further substituted with one or two R 3a or (i1) C3-C6 cycloalkyl substituted with phenyl (wherein the phenyl is selected from the group consisting of LG, ... 1 and said phenyl is further optionally substituted with one, two or three R 3a groups, and the cycloalkyl may optionally be substituted with one or two R 3a and optionally substituted with a group. or a salt thereof, and / or a stereoisomer or mixture of stereoisomers, provided that said compound is methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate

[0178] [ka]

[0179] or a salt thereof, and / or a stereoisomer or mixture of stereoisomers thereof) is provided.

[0180] LG 1 may be, for example, a suitable leaving group in a suitable nucleophilic aromatic substitution or a suitable cross-coupling (but not limited to Suzuki-Miyaura coupling).

[0181] In one or more embodiments, a) a compound of formula (A):

[0182] [ka]

[0183] The compound of R 2 '-C(O)H; or b) contacting a compound of formula (B):

[0184] [ka]

[0185] The compound of R 2 '-C(NH)NH2, where R 20 is Me or CD3); or c) a compound of formula (C):

[0186] [ka]

[0187] The compound of R 2 '-H, where LG 1 is fluoro, chloro, bromo, iodo, triflate, mesylate, triazole, pyrazole, boronic acid, boronic ester, or allyl trifluoroborate; and optionally isolating the compound of formula (I); 2 (b1) phenyl substituted with 5- or 6-membered heteroaryl, wherein the 5- or 6-membered heteroaryl is LG 1 and optionally 1, 2 or 3 R 3a groups, and the phenyl may further optionally be substituted with one, two or three R 3a (c1) optionally substituted with 1, 2 or 3 R 3a and may be substituted with -phenyl-LG 1 phenyl substituted with -phenyl-LG 1 The phenyl in the group may optionally be substituted with one, two or three R 3a (e1) optionally substituted with 1, 2 or 3 R 3a and may be substituted with -phenyl-LG 1 5- or 6-membered heteroaryl substituted with 1, 2, or 3 R 3a (f1) optionally substituted with 1, 2 or 3 R 3a and may be substituted with a -(5- or 6-membered heteroaryl)-LG1 5- or 6-membered heteroaryl substituted with -(5- or 6-membered heteroaryl)-LG 1 The 5- or 6-membered heteroaryl in 3a (h1) substituted with NH or OH and optionally further substituted with one or two R 3a or (i1) C3-C6 cycloalkyl substituted with phenyl (wherein the phenyl is selected from the group consisting of LG, ... 1 and said phenyl is further optionally substituted with one, two or three R 3a groups, and the cycloalkyl may optionally be substituted with one or two R 3a and (b) a methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate group;

[0188] [ka]

[0189] or a salt thereof and / or a stereoisomer or mixture of stereoisomers thereof). In one or more embodiments, the method of making a compound of Formula (I) includes contacting under basic conditions.

[0190] In one or more embodiments of Formula (I), the compound is selected from any of compounds 1-44 from Table 1, or a pharmaceutically acceptable salt thereof. In one or more embodiments of Formula (I), the compound is selected from any of compounds 1-44 from Table 1, or an isomer thereof. In one or more embodiments of Formula (I), the compound is selected from any of compounds 1-44 from Table 1, or a single stereoisomer of a mixture of stereoisomers thereof, and / or a pharmaceutically acceptable salt thereof. In one or more embodiments of Formula (I), the compound is selected from any of compounds 1-44 from Table 1, or a prodrug, pharmaceutically acceptable salt, metabolite, polymorph, solvate, hydrate, stereoisomer, or tautomer thereof.

[0191] [Table 1]

[0192] JPEG2025535316000037.jpg212117

[0193] JPEG2025535316000038.jpg230116

[0194] JPEG2025535316000039.jpg229117

[0195] JPEG2025535316000040.jpg229116

[0196] JPEG2025535316000041.jpg221117

[0197] JPEG2025535316000042.jpg206117

[0198] JPEG2025535316000043.jpg141117

[0199] In one or more embodiments, the compound has the formula:

[0200] [ka]

[0201] A compound or a salt thereof according to any one of (In the formula, LG 1 is a leaving group such as fluoro, chloro, bromo, iodo, triflate, mesylate, triazole, pyrazole, boronic acid, boronic ester, or allyltrifluoroborate; R 1 is hydrogen, deuterium, C1-C3 alkyl, —OH, —O—C1-C3 alkyl, —CH2OH, or —B(OH)2; R 1a is hydrogen, deuterium, or C1-C3 alkyl; R 20 is alkyl, preferably methyl or ethyl, or CD3; R 2 'teeth, (b1) phenyl substituted with 5- or 6-membered heteroaryl, wherein the 5- or 6-membered heteroaryl is LG 1 and optionally 1, 2 or 3 R 3a groups, and the phenyl may further optionally be substituted with one, two or three R 3a optionally substituted with a group, (c1) optionally 1, 2 or 3 R 3a and may be substituted with -phenyl-LG 1 phenyl substituted with -phenyl-LG 1 The phenyl in the group may optionally be substituted with one, two or three R 3a optionally substituted with a group); (e1) optionally 1, 2 or 3 R 3a and may be substituted with -phenyl-LG 15- or 6-membered heteroaryl substituted with 1, 2, or 3 R 3a optionally substituted with a group); (f1) optionally 1, 2 or 3 R 3a and may be substituted with a -(5- or 6-membered heteroaryl)-LG 1 5- or 6-membered heteroaryl substituted with -(5- or 6-membered heteroaryl)-LG 1 The 5- or 6-membered heteroaryl in 3a optionally substituted with a group); (h1) substituted with NH2 or OH and optionally further substituted with one or two R 3a C3-C6 cycloalkyl optionally substituted with a group; or (i1) C3-C6 cycloalkyl substituted with phenyl (wherein the phenyl is LG 1 and said phenyl is further optionally substituted with one, two or three R 3a groups, and the cycloalkyl may optionally be substituted with one or two R 3a (which may be substituted with a group) is); (However, the compound is: Methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate

[0202] [ka]

[0203] or its salts or enantiomers) is provided herein.

[0204] In one or more embodiments, the compound is selected from any of the following compounds A1-A6 from Table 2, or a prodrug, pharmaceutically acceptable salt, metabolite, polymorph, solvate, hydrate, stereoisomer, or tautomer thereof, and the compound is useful for making at least a compound of Formula (I).

[0205] [Table 2]

[0206] In some or all embodiments: (a) compounds described herein, e.g., compounds of formula (I), and compounds 1-44, and pharmaceutically acceptable salts and compositions thereof; (b) a compound described herein, e.g., a compound of formula (I), and compounds 1-44, and pharmaceutically acceptable salts and compositions thereof, for use in stimulating wound regeneration in a mammal; (c) compounds described herein, e.g., compounds of formula (I), and compounds 1-44, and pharmaceutically acceptable salts and compositions thereof, for use in inhibiting Wnt transcripts or Wnt signaling pathway activity; (d) methods for preparing compounds of formula (I), as described herein, for example, compounds 1-44, as described in more detail elsewhere herein; (e) a pharmaceutical formulation comprising a compound described herein, e.g., a compound of Formula (I), and compounds 1-44, or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier (e.g., a diluent); (f) a method for treating a condition associated with Wnt transcript or Wnt signaling pathway activity in a mammal, comprising administering a therapeutically effective amount of a compound described herein, e.g., a compound of formula (I), and compounds 1-44, or a pharmaceutically acceptable salt or composition thereof; (g) a method for treating a wound in a mammal, comprising administering a therapeutically effective amount of a compound described herein, e.g., a compound of formula (I), and compounds 1-44, or a pharmaceutically acceptable salt or composition thereof; (h) Pharmaceutical formulations comprising a compound described herein, e.g., a compound of formula (I), and compounds 1-44, or a pharmaceutically acceptable salt thereof, together with one or more other active agents for treating wounds and / or conditions modulated by Wnt transcripts or Wnt signaling pathway activity, optionally in a pharmaceutically acceptable carrier (e.g., a diluent); (i) a method for treating a wound in a mammal, comprising administering a therapeutically effective amount of a compound described herein, e.g., a compound of formula (I), and compounds 1-44, or a pharmaceutically acceptable salt or composition thereof, in combination with and / or in place of one or more agents for treating wounds and / or conditions modulated by Wnt transcripts or Wnt signaling pathway activity; (j) a method for treating a condition associated with Wnt transcript or Wnt signaling pathway activity in a mammal, comprising administering a therapeutically effective amount of a compound described herein, e.g., a compound of formula (I), and compounds 1-44, or a pharmaceutically acceptable salt or composition thereof, in combination with and / or in place of one or more agents for treating wounds; and (k) Use of any compound described herein, e.g., compounds of formula (I), and compounds 1-44, or a composition comprising any compound described herein, e.g., compounds of formula (I), and compounds 1-44, or a pharmaceutically acceptable salt thereof, to treat a wound associated with Wnt transcript or Wnt signaling pathway activity as described herein, optionally in combination with and / or in place of one or more agents for treating the wound. is provided herein.

[0207] optically active compound It is understood that the compounds provided herein have several chiral centers and can exist and be isolated in optically active and racemic forms. It is understood that any racemic, optically active, diastereomeric, tautomeric, stereoisomeric, mixtures, or combinations thereof of the compounds provided herein that possess the useful properties described herein are within the scope of the present invention. Methods for preparing optically active forms (in some or all embodiments, by resolution of racemic forms by recrystallization techniques, synthesis from optically active starting materials, chiral synthesis, or chromatographic separation using chiral stationary phases) are well known in the art.

[0208] Methods for obtaining optically active materials in some or all embodiments are known in the art and include at least the following: i) Physical separation of crystals - a technique for manually separating macroscopic crystals of individual stereoisomers. This technique can be used when crystals of separate stereoisomers are present, i.e., when the material is a conglomerate and the crystals are visually distinct; ii) simultaneous crystallization - a technique for crystallizing individual stereoisomers separately from a solution of the racemate, possible only if the latter are solid-state aggregates; iii) Enzymatic separation - a technique that utilizes the different reaction rates of stereoisomers with enzymes to partially or completely separate the racemate; iv) enzymatic asymmetric synthesis - a synthetic technique in which at least one step of the synthesis uses an enzymatic reaction to obtain a stereomerically pure or enriched synthetic precursor of a desired stereoisomer; v) Chemical asymmetric synthesis - a synthetic technique in which a desired stereoisomer is synthesized from an achiral precursor under conditions that result in asymmetry (i.e., chirality) in the product, which may be achieved by using a chiral catalyst or chiral auxiliary; vi) Diastereomeric separation - a technique in which a racemic compound is reacted with an enantiomerically pure reagent (chiral auxiliary) to convert the individual enantiomers into diastereomers. The resulting diastereomers are separated by chromatography or crystallization due to their more distinct structural differences, and the chiral auxiliary is subsequently removed to give the desired enantiomer; vii) Primary and secondary asymmetric transformations - techniques in which the diastereomers from the racemate equilibrate to a point where the diastereomer derived from the desired enantiomer predominates in solution, or the preferential crystallization of the diastereomer derived from the desired enantiomer disrupts the equilibrium, ultimately converting in principle all of the material to the crystalline diastereomer derived from the desired enantiomer, which is then released from the diastereomer; viii) Kinetic resolution - this technique refers to achieving partial or complete resolution of a racemate (or further resolution of a partially resolved compound) by the unequal reaction rates of stereoisomers with chiral non-racemic reagents or catalysts under kinetic conditions; ix) stereospecific synthesis from non-racemic precursors - a synthetic technique in which the desired stereoisomer is obtained from non-chiral starting materials and in which the stereochemical integrity is not or only minimally compromised during the synthesis; x) Chiral liquid chromatography - a technique in which stereoisomers of a racemate are separated in a liquid mobile phase by their different interactions with the stationary phase. The stationary phase can be made from a chiral substance, or the mobile phase can contain additional chiral substances to cause different interactions; xi) Chiral gas chromatography - a technique in which the racemate is volatilized and stereoisomers are separated by differences in their interactions in the gaseous mobile phase using a column containing a fixed non-racemic chiral adsorbent phase; xii) Chiral solvent extraction - a technique for separating stereoisomers by preferentially dissolving one stereoisomer in a particular chiral solvent; xiii) Transport across chiral membranes - a technique in which a racemate is contacted with a thin membrane barrier. The barrier typically separates two miscible fluids (one containing the racemate), and a driving force such as concentration or pressure differential causes preferential transport across the membrane barrier. Separation occurs as a result of the non-racemic chiral nature of the membrane, which allows only one stereoisomer of the racemate to pass through.

[0209] In some or all embodiments, compositions of compounds are provided that comprise a substantially pure designated stereoisomer of a compound. In some or all of the methods and compounds, the compound is substantially free of other stereoisomers. In some or all embodiments, the composition comprises at least 85%, 90%, 95%, 98%, 99%, or 100% by weight of a compound that is the designated stereoisomer, with the remainder comprising other species or stereoisomers.

[0210] Isotopically enriched compounds Isotopically enriched compounds are also provided herein.

[0211] Isotopic enrichment (deuteration in some or all embodiments) of pharmaceuticals to improve pharmacokinetics ("PK"), pharmacodynamics ("PD"), and toxicity profiles has previously been demonstrated for several classes of drugs. For example, Lijinsky et. al., Food Cosmet. Toxicol., 20: 393 (1982); Lijinsky et. al., J. Nat. Cancer Inst., 69: 1127 (1982); Mangold et. al., Mutation Res. 308: 33 (1994); Gordon et. al., Drug Metab. Dispos., 15: 589 (1987); Zello et. al., Metabolism, 43: 487 (1994); Gately et. al., J. Nucl. Med., 27: 388 (1986); Wade D, Chem. Biol. Interact. 117: 191 (1999).

[0212] Isotopic enrichment of a drug may be used in some or all embodiments to (1) reduce or eliminate unwanted metabolites, (2) increase the half-life of the parent drug, (3) decrease the number of doses required to achieve a desired effect, (4) decrease the dose required to achieve a desired effect, (5) increase the formation of active metabolites, if formed, and / or (6) decrease the production of harmful metabolites in specific tissues, and / or to make the drug more effective and / or safer for combination therapy, whether or not the combination therapy is intended.

[0213] Replacing an atom with one of its isotopes often changes the reaction rate of a chemical reaction. This phenomenon is known as the kinetic isotope effect ("KIE"). For example, if a C-H bond is broken in the rate-dependent step of a chemical reaction (i.e., the step with the highest transition state energy), replacing the hydrogen with deuterium will reduce the reaction rate and slow down the process. This phenomenon is known as the deuterium kinetic isotope effect ("DKIE"). See, for example, Foster et al., Adv. Drug Res., vol. 14, pp. 1-36 (1985); Kushner et al., Can. J. Physiol. Pharmacol., vol. 77, pp. 79-88 (1999).

[0214] The magnitude of the DKIE can be expressed as the ratio between the rate of a given reaction in which a C-H bond is broken and the rate of the same reaction when hydrogen is replaced by deuterium. The DKIE can range from approximately 1 (no isotope effect) to very large values, such as over 50, which means that the reaction slows by more than 50 times when hydrogen is replaced by deuterium. High DKIE values ​​may be due in part to a phenomenon known as tunneling, a consequence of the uncertainty principle. Tunneling occurs because the small mass of the hydrogen atom allows transition states involving a proton to occasionally occur without the necessary activation energy. Because deuterium is more massive than hydrogen, this phenomenon is statistically much less likely to occur.

[0215] Tritium ("T") is a radioactive isotope of hydrogen used in research, fusion reactors, neutron generators, and radiopharmaceuticals. Tritium is a hydrogen atom with two neutrons in its nucleus and an atomic mass close to 3. It occurs naturally in the environment in very low concentrations, most commonly found as T2O. Tritium decays slowly (half-life = 12.3 years), emitting low-energy beta particles that cannot penetrate the outer layer of human skin. Internal exposure is the primary hazard associated with this isotope, but large amounts must be ingested to pose a significant health risk. Compared to deuterium, less tritium must be ingested to reach dangerous levels. Substituting tritium ("T") for hydrogen results in a stronger bond than deuterium, resulting in a numerically larger isotope effect. Similarly, the substitution of carbon for tritium ("T") results in a stronger bond than deuterium, resulting in a numerically larger isotope effect. 13 C or 14 C, sulfur 33 S, 34 S or 36 S, nitrogen 15 N and oxygen 17 O or 18 Substitution with isotopes of other elements, including but not limited to O, may result in similar kinetic isotope effects.

[0216] For example, DKIEs have been used to reduce the hepatotoxicity of halothane, presumably by limiting the production of reactive species such as trifluoroacetyl chloride. However, this method is not applicable to all drugs. For example, deuterium incorporation can lead to metabolic switching. The concept of metabolic switching proposes that when a xenogen is sequestered by a phase I enzyme, it may bind transiently and rebind in various conformations before chemical reaction (e.g., oxidation). This hypothesis is supported by the relatively large size of the binding pockets of many phase I enzymes and the promiscuous nature of many metabolic reactions. Metabolic switching can potentially lead to different ratios of known and entirely new metabolites. This new metabolic profile can result in greater or lesser toxicity.

[0217] In some embodiments, the compounds described herein may be used as radiopharmaceuticals, such as imaging agents. In one example, the radiopharmaceutical is a positron emission tomography (PET) imaging agent. In such embodiments, substitution of atoms in the compound with radionuclides (e.g., positron-emitting isotopes) allows for the synthesis of radiopharmaceuticals that can function as imaging agents. In some embodiments, radionuclides that can be substituted in the compounds described herein include: 18 F, 11 C. 13 N, 15 O. 76 Br, and 124 I. In some embodiments, the compound is isotopically enriched at one or more atoms, one atom, two atoms, or three atoms. In some embodiments, the compound is administered as an isotopic composition.

[0218] Animal bodies express a variety of enzymes for the purpose of eliminating foreign substances, such as therapeutic drugs, from their circulatory systems. In some or all embodiments, such enzymes include cytochrome P450 enzymes ("CYP"), esterases, proteases, reductases, dehydrogenases, and monoamine oxidases to react with these foreign substances and convert them into more polar intermediates or metabolites for renal excretion. Some of the most common metabolic reactions of pharmaceutical compounds involve the oxidation of carbon-hydrogen (CH) bonds to either carbon-oxygen (CO) bonds or carbon-carbon (CC) pi bonds. The resulting metabolites may be stable or unstable under physiological conditions and may have significantly different pharmacokinetic, pharmacodynamic, and acute and long-term toxicity profiles compared to the parent compound. For many drugs, such oxidation is rapid. As a result, these drugs often need to be administered multiple times daily or in high doses.

[0219] Thus, isotopic enrichment at certain positions in the compounds provided herein results in a detectable KIE and affects the pharmacokinetic, pharmacological, and / or toxicological profiles of the compounds provided herein compared to analogous compounds with natural isotopic composition.

[0220] Compound production The compounds provided herein can be prepared, isolated, or obtained by any method apparent to one skilled in the art. The compounds provided herein can be prepared according to the exemplary preparation schemes provided below. Reaction conditions, steps, and reactants not provided in the exemplary preparation schemes will be apparent and known to one skilled in the art.

[0221] Additional steps and reagents not provided in the exemplary preparation schemes will be known to those skilled in the art. For example, intermediates and compounds can be prepared using procedures known to those skilled in the art or as disclosed in U.S. Provisional Application Nos. 63 / 417,257; 63 / 418,947; and 63 / 418,956 (the synthetic methods disclosed therein are incorporated by reference in their entirety). Exemplary preparation methods are described in detail in the Examples herein.

[0222] In one or more embodiments, a) a compound of formula (A):

[0223] [ka]

[0224] The compound of R 2 '-C(O)H; or b) contacting a compound of formula (B):

[0225] [ka]

[0226] The compound of R 2 '-C(NH)NH2, where R 20 is Me or CD3); or c) a compound of formula (C):

[0227] [ka]

[0228] The compound of R 2 '-H, where LG 1 is fluoro, chloro, bromo, iodo, triflate, mesylate, triazole, pyrazole, boronic acid, boronic ester, or allyl trifluoroborate; and optionally isolating the compound of formula (I); 2(b1) phenyl substituted with 5- or 6-membered heteroaryl, wherein the 5- or 6-membered heteroaryl is LG 1 and optionally 1, 2 or 3 R 3a groups, and the phenyl may further optionally be substituted with one, two or three R 3a (c1) optionally substituted with 1, 2 or 3 R 3a and may be substituted with -phenyl-LG 1 phenyl substituted with -phenyl-LG 1 The phenyl in the group may optionally be substituted with one, two or three R 3a (e1) optionally substituted with 1, 2 or 3 R 3a and may be substituted with -phenyl-LG 1 5- or 6-membered heteroaryl substituted with 1, 2, or 3 R 3a (f1) optionally substituted with 1, 2 or 3 R 3a and may be substituted with a -(5- or 6-membered heteroaryl)-LG 1 5- or 6-membered heteroaryl substituted with -(5- or 6-membered heteroaryl)-LG 1 The 5- or 6-membered heteroaryl in 3a (h1) substituted with NH or OH and optionally further substituted with one or two R 3a or (i1) C3-C6 cycloalkyl substituted with phenyl (wherein the phenyl is selected from the group consisting of LG, ... 1 and said phenyl is further optionally substituted with one, two or three R 3a groups, and the cycloalkyl may optionally be substituted with one or two R 3a and optionally substituted with a group. 1. A method for preparing a compound of formula (I), comprising the step of:

[0229] [ka]

[0230] or a salt thereof and / or a stereoisomer or mixture of stereoisomers thereof) are provided. In one or more embodiments, the method for preparing a compound of Formula (I) comprises contacting under basic conditions; and all other groups are as defined herein in all embodiments; and optionally, R 1 and R 1a is independently hydrogen or alkyl.

[0231] Those skilled in the art will understand that the order of steps of any method described herein may be varied. Other variations will be apparent to those skilled in the art, and all such variations are contemplated within the scope of the embodiments presented herein.

[0232] Pharmaceutical compositions and methods of administration The compounds provided herein can be formulated into pharmaceutical compositions using methods available in the art and methods disclosed herein.Any of the compounds disclosed herein can be provided in a suitable pharmaceutical composition and administered by a suitable route.Provided herein is a pharmaceutical composition comprising a compound of formula (I) described in any and all embodiments herein and a pharmaceutically acceptable carrier.

[0233] In some embodiments, the composition is a topical composition.

[0234] The methods provided herein involve administering a pharmaceutical composition containing at least one compound described herein (including a compound of Formula (I) in salt form, where appropriate), alone or in combination with one or more compatible pharmaceutically acceptable carriers, such as diluents or adjuvants, or other medicinal agents, for the treatment of wounds and / or conditions modulated by Wnt transcripts or Wnt signaling pathway activity.

[0235] The compositions described herein (including GO-HA pharmaceutical compositions) can include other medicinal or therapeutic compounds in addition to the compound of Formula (I). In other words, the composition (including GO-HA pharmaceutical compositions) in which the compound of Formula (I) is present can function as a base dispersion medium into which other medicinal or therapeutic agents (particularly those that are hydrophobic) can be dispersed for topical administration, for example, to a wound. These agents include antifibrotic compounds such as pirfenidone, halofuginone, nintedanib, tocilizumab, rilonacept, anticancer agents, anti-inflammatory agents, analgesics, antibiotics, Wnt inhibitors, hedgehog pathway inhibitors, TGF-β inhibitors, LOX inhibitors, and the like.

[0236] In some embodiments, the compositions may include a second wound medication or treatment, including one or more of a corticosteroid, a cytotoxic drug, an antibiotic, an antiseptic, nicotine, an antiplatelet drug, an NSAID, colchicine, an anticoagulant, a vasoconstrictor or immunosuppressant, a growth factor, an antibody, a protease, a protease inhibitor, an antimicrobial peptide, an adhesion peptide, a hemostatic agent, a live cell, honey, or nitric oxide. These treatments may be delivered in a dosage form separate from the compositions described herein, or may be included as an additional component of the compositions described herein and thus delivered along with a compound of Formula (I) (or any embodiment thereof, including in some embodiments Compound 1, Compound 7, or Compound 8).

[0237] In some or all embodiments, a second drug may be formulated or packaged together with the compound provided herein. Of course, the second drug will be formulated with the compound provided herein only if, according to the judgment of one skilled in the art, such co-formulation does not interfere with the activity or administration method of either drug. In some or all embodiments, the compound provided herein and the second drug are formulated separately. They may be packaged together or separately, for the convenience of one skilled in the art.

[0238] In practice, the active agents provided herein may be administered by any conventional route, in particular parenterally, rectally, orally, by inhalation (eg, in the form of an aerosol), or topically.

[0239] The composition(s) of the present disclosure described herein may be administered by topically applying the composition(s) to a wound. When the composition is contained in a medical device described herein that includes a substrate such as a patch or pad, the medical device may be secured to the wound so that the composition contacts the wound.

[0240] As solid compositions for oral administration, tablets, pills, hard gelatin capsules, powders or granules can be used, in which the active product is mixed with one or more inert diluents or adjuvants, such as sucrose, lactose or starch.

[0241] These compositions may contain substances other than diluents, for example lubricants such as magnesium stearate, or coatings intended to control release.

[0242] As liquid compositions for oral administration, pharmaceutically acceptable solutions, suspensions, emulsions, syrups and elixirs containing inert diluents such as water or liquid paraffin may be used. These compositions may also contain substances other than diluents, in some or all embodiments, wetting, sweetening or flavoring products.

[0243] Compositions for parenteral administration may be emulsions or sterile solutions. As solvents or vehicles, propylene glycol, polyethylene glycol, vegetable oils, especially olive oil, or injectable organic esters, in some or all embodiments, ethyl oleate, may be used. These compositions may also contain adjuvants, especially wetting agents, isotonicity adjusting agents, emulsifying agents, dispersing agents, and stabilizing agents. Sterilization can be carried out in several ways, and in some or all embodiments, it can be carried out using a bacteriological filter, by irradiation, or by heat. They can also be produced in the form of sterile solid compositions, which can be dissolved in sterile water or any other injectable sterile medium at the time of use.

[0244] Compositions for rectal administration are suppositories or rectal capsules which contain, in addition to the active ingredient, excipients such as cocoa butter, semi-synthetic glycerides or polyethylene glycols.

[0245] The composition can also be an aerosol.When used in the form of a liquid aerosol or spray, the composition can be a stable sterile solution or a solid composition that is dissolved in non-pyrogenic sterile water, saline or other pharmaceutically acceptable vehicle when used.When used in the form of a dry aerosol intended for direct inhalation, the active ingredient is finely divided and combined with a water-soluble solid diluent or vehicle, in some or all embodiments, dextran, mannitol or lactose.In one or more embodiments, the pharmaceutical composition provided herein is a spray.

[0246] In some or all embodiments, the compositions provided herein are pharmaceutical compositions or single unit dosage forms. The pharmaceutical compositions and single unit dosage forms provided herein comprise a therapeutically effective amount of one or more therapeutic agents (e.g., a compound provided herein or other therapeutic agents) and typically one or more pharmaceutically acceptable carriers (e.g., excipients). In certain embodiments, and in this context, the term "pharmaceutically acceptable" means approved by a federal or state regulatory agency or listed in the United States Pharmacopoeia or other generally recognized pharmacopoeias for use in animals, particularly humans. In some embodiments, the term "carrier" includes a diluent, disintegrant, lubricant, adjuvant (e.g., Freund's adjuvant (complete and incomplete)), excipient, or vehicle with which the therapeutic agent is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils (including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc.). When pharmaceutical compositions are administered intravenously, water can be used as a carrier.Saline, aqueous glucose solution and aqueous glycerol solution can also be used as liquid carriers, particularly for injections.Examples of suitable pharmaceutical carriers are listed in Remington: The Science and Practice of Pharmacy; Pharmaceutical Press; 22nd Edition (September 15, 2012).

[0247] Typical pharmaceutical compositions and dosage forms contain one or more excipients. Suitable excipients are well known to those skilled in the art of pharmacy, and suitable excipients in some or all embodiments include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, etc. Whether a particular excipient is suitable for incorporation into a pharmaceutical composition or dosage form depends on various factors well known in the art, including, but not limited to, the manner in which the dosage form will be administered to a mammal and the specific active ingredients in the dosage form. The composition or single-unit dosage form may also contain minor amounts of wetting or emulsifying agents, or pH buffering agents, if desired.

[0248] The lactose-free compositions provided herein are well known in the art and may, in some or all embodiments, contain excipients listed in the United States Pharmacopoeia (USP 36-NF 31 S2). Generally, lactose-free compositions contain an active ingredient, a binder / filler, and a lubricant in pharmaceutically compatible and pharmaceutically acceptable amounts. An exemplary lactose-free dosage form contains an active ingredient, microcrystalline cellulose, pregelatinized starch, and magnesium stearate.

[0249] Because water can accelerate the decomposition of some compounds, anhydrous pharmaceutical compositions and dosage forms containing active ingredients are also encompassed herein. For example, the addition of water (e.g., 5%) is widely accepted in the field as a means of simulating long-term storage to determine properties such as shelf life or stability over time of a formulation. See, for example, Jens T. Carstensen, Drug Stability: Principles & Practice, 2nd Edition, Marcel Dekker, New York, 1995, pp. 379-80. In fact, water and heat accelerate the decomposition of some compounds. Therefore, the effect of water on a formulation can be very important, since moisture and / or humidity are commonly encountered during the manufacture, handling, packaging, storage, shipping, and use of formulations.

[0250] Anhydrous pharmaceutical compositions and dosage forms provided herein can be prepared using anhydrous or low moisture containing ingredients and low moisture or low humidity conditions. Pharmaceutical compositions and dosage forms comprising lactose and at least one active ingredient comprising a primary or secondary amine can be anhydrous if substantial contact with moisture and / or humidity is expected during manufacturing, packaging, and / or storage.

[0251] An anhydrous pharmaceutical composition should be prepared and stored such that its anhydrous nature is maintained. Accordingly, anhydrous compositions may be packaged using materials known to prevent exposure to water such that they can be included in suitable formulary kits. In some or all embodiments, suitable packaging includes, but is not limited to, hermetically sealed foils, plastics, unit dose containers (e.g., vials), blister packs, and strip packs.

[0252] Pharmaceutical compositions and dosage forms that comprise one or more compounds that reduce the rate at which the active ingredient decomposes are further provided. Such compounds, which are referred to herein as "stabilizers," include, but are not limited to, antioxidants such as ascorbic acid, pH buffers, or salt buffers.

[0253] Pharmaceutical compositions and dosage forms are further provided that include one or more chemical permeation enhancers, which in some or all embodiments include, but are not limited to, ethanol, amides (such as ozone and laurocapram), alkyl and benzoic acid esters, fatty acid esters (such as isopropyl myristate, propylene glycol monocaprylate, and propylene glycomonolaurate), Transcutol®, fatty acids (oleic acid), glycols, pyrrolidones (such as N-methyl-2-pyrrolidone and 2-pyrrolidone, dimethyl sulfoxide (DMSO), terpenes (such as essential oils containing terpenes), phospholipids, and / or cyclodextrins.

[0254] Pharmaceutical compositions and unit dosage forms can take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained-release formulations, and the like. Oral formulations may include standard carriers such as pharmaceutical grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, and the like. Such compositions and dosage forms contain a prophylactically or therapeutically effective amount of a prophylactic or therapeutic agent, in some or all embodiments, in purified form, together with an appropriate amount of carrier to provide a form for proper administration to a mammal. The formulation should suit the mode of administration. In some or all embodiments, the pharmaceutical composition or unit dosage form is sterile and in a form suitable for administration to mammals, and in some or all embodiments, humans.

[0255] A pharmaceutical composition is formulated to be compatible with its intended route of administration. In some or all embodiments, routes of administration include, but are not limited to, parenteral, e.g., intraspinal, epidural, local or regional for peripheral nerve block, intravenous, intradermal, subcutaneous, intramuscular, subcutaneous, oral, buccal, sublingual, inhalation, intranasal, transdermal, topical (including ophthalmic and, in some embodiments, corneal), transmucosal, intratumoral, intrasynovial, and rectal administration. In certain embodiments, the composition is routinely formulated as a pharmaceutical composition suitable for intravenous, subcutaneous, intramuscular, oral, intranasal, or topical (including ophthalmic and, in some embodiments, corneal) administration to humans. In certain embodiments, the pharmaceutical composition is routinely formulated for subcutaneous administration to humans. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. Optionally, the composition may also include a solubilizing agent and a local anesthetic, such as lignocamune, to ease pain at the injection site.

[0256] In some or all embodiments, dosage forms include, but are not limited to, sprays, tablets; caplets; capsules, such as soft elastic gelatin capsules; cachets; troches; lozenges; dispersions; suppositories; ointments; poultices (coulds); pastes; powders; dressings; creams; plasters; solutions; patches; aerosols (e.g., nasal sprays or inhalers); gels; liquid dosage forms suitable for oral or mucosal administration to a mammal, including suspensions (e.g., aqueous or non-aqueous suspensions, oil-in-water emulsions, or water-in-oil liquid emulsions), solutions, and elixirs; liquid dosage forms suitable for parenteral administration to a mammal; and sterile solids (e.g., crystalline or amorphous solids) that can be reconstituted to provide liquid dosage forms suitable for parenteral administration to a mammal.

[0257] The composition, shape, and type of dosage forms provided herein typically vary depending on their intended use. In some or all embodiments, a dosage form used for the initial treatment of a disease, disorder, or condition may contain larger amounts of one or more active ingredients than a dosage form used for the maintenance treatment of the same disease, disorder, or condition. Similarly, a parenteral dosage form may contain smaller amounts of one or more active ingredients than an oral dosage form used to treat the same disease or disorder. These and other ways in which specific dosage forms encompassed herein differ from one another will be readily apparent to those skilled in the art. See, e.g., Remington: The Science and Practice of Pharmacy; Pharmaceutical Press; 22nd Edition (September 15, 2012).

[0258] Generally, the ingredients of the composition are supplied, in some or all embodiments, either separately or mixed together in unit dosage form as a dry lyophilized powder or water-free concentrate in a hermetically sealed container such as an ampoule or sachet indicating the quantity of active agent. Where the composition is administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical-grade water or saline. Where the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients can be mixed prior to administration.

[0259] Typical dosage forms comprise a compound provided herein, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, in the range of about 0.1 mg to about 1000 mg per day, administered once daily in the morning or in divided doses throughout the day with meals. Particular dosage forms may have about 0.1, 0.2, 0.3, 0.4, 0.5, 1.0, 2.0, 2.5, 5.0, 10.0, 15.0, 20.0, 25.0, 50.0, 100, 200, 250, 500, or 1000 mg of active compound.

[0260] Oral dosage form Pharmaceutical compositions suitable for oral administration may be presented as discrete dosage forms, such as, but not limited to, tablets (e.g., chewable tablets), caplets, capsules, and liquids (e.g., flavored syrups). Such dosage forms contain a predetermined amount of active ingredient and can be prepared by methods of pharmacy well known to those skilled in the art. See generally, Remington: The Science and Practice of Pharmacy; Pharmaceutical Press; 22nd Edition (September 15, 2012).

[0261] In some or all embodiments, the oral dosage forms are solid and are manufactured under anhydrous conditions using anhydrous ingredients, as described in detail herein. However, the scope of the compositions provided herein extends beyond anhydrous solid oral dosage forms. As such, additional forms are described herein.

[0262] Typical oral dosage forms are prepared by intimately mixing the active ingredient with at least one excipient according to conventional pharmaceutical compounding techniques. The excipients can take a variety of forms depending on the dosage form desired for administration. In some or all embodiments, excipients suitable for use in oral liquid or aerosol dosage forms include, but are not limited to, water, glycols, oils, alcohols, flavoring agents, preservatives, and coloring agents. In some or all embodiments, excipients suitable for use in solid oral dosage forms (e.g., powders, tablets, capsules, and caplets) include, but are not limited to, starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, and disintegrants.

[0263] Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit forms, in which case solid excipients are employed. If desired, tablets may be coated by standard aqueous or nonaqueous techniques. Such dosage forms may be prepared by any of the methods of pharmacy. In general, pharmaceutical compositions and dosage forms are prepared by uniformly and intimately admixing the active ingredients with liquid carriers, finely divided solid carriers, or both, and then, if necessary, shaping the product into the desired presentation.

[0264] In some or all embodiments, tablets may be prepared by compression or molding. Compressed tablets may be prepared by compressing the active ingredient in a free-flowing form such as powder or granules, optionally mixed with an excipient, in a suitable machine. Molded tablets may be prepared by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

[0265] In some or all embodiments, one or more of the following may be used in the pharmaceutical composition: benzyl alcohol, butylparaben, butylated hydroxytoluene, calcium carbonate, candelilla wax, colloidal silicon dioxide, calcium stearate, calcium disodium EDTA, copolyvidone or copovidone, calcium hydrogen phosphate dihydrate, crospovidone, calcium phosphate (di- and tribasic), emollient (glyceryl monostearate), iron oxide ivy yellow, and iron.

[0266] In some or all embodiments, excipients that may be used in oral dosage forms include, but are not limited to, binders, fillers, disintegrants, and lubricants. Suitable binders for use in pharmaceutical compositions and dosage forms include, but are not limited to, copolyvidone or copovidone, corn starch, potato starch, or other starches, gelatin, natural and synthetic gums such as acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, calcium carboxymethylcellulose, sodium carboxymethylcellulose), polyvinylpyrrolidone, methylcellulose, pregelatinized starch, hydroxypropyl methylcellulose (e.g., Nos. 2208, 2906, 2910), microcrystalline cellulose, and mixtures thereof.

[0267] In some or all embodiments, fillers suitable for use in the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof. The binder or filler in a pharmaceutical composition is typically present in from about 50 to about 99 weight percent of the pharmaceutical composition or dosage form.

[0268] In some or all embodiments, suitable forms of microcrystalline cellulose include, but are not limited to, materials sold as AVICEL PH 101, AVICEL PH 103, AVICEL RC 581, AVICEL PH 105 (FMC Corporation, American Viscose Division, Avicel Sales, Marcus Hook, PA), and mixtures thereof. A specific binder is a mixture of microcrystalline cellulose and sodium carboxymethylcellulose sold as AVICEL RC 581. Suitable anhydrous or low moisture excipients or additives include AVICEL PH 103, AVICEL PH 105, AVICEL RC 581, AVICEL PH 105 ... TM and Starch 1500 LM.

[0269] Disintegrants are used in compositions to provide tablets that disintegrate when exposed to an aqueous environment. Tablets containing too much disintegrant may disintegrate during storage, while those containing too little may not disintegrate at the desired rate or under the desired conditions. Therefore, to form a solid oral dosage form, a sufficient amount of disintegrant should be used, but not too much, nor too little, to adversely alter the release of the active ingredient. The amount of disintegrant used will vary depending on the type of formulation and is readily discernible to those skilled in the art. Typical pharmaceutical compositions contain about 0.5 to about 15% by weight of disintegrant, particularly about 1 to about 5% by weight.

[0270] Disintegrants that can be used in pharmaceutical compositions and dosage forms include, but are not limited to, crospovidone, agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, pregelatinized starch, other starches, clays, other algins, other celluloses, gums, and mixtures thereof.

[0271] Lubricants that may be used in pharmaceutical compositions and dosage forms include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, and mixtures thereof. Additional lubricants, in some or all embodiments, include syloid silica gel (AEROSIL 200, manufactured by W.R. Grace Co., Baltimore, Maryland), coagulated aerosol of synthetic silica (available from Degussa Co., Plano, Texas), CAB O SIL (a pyrogenic silicon dioxide product available from Cabot Co., Boston, Massachusetts), and mixtures thereof. If used at all, lubricants are typically used in an amount of less than about 1 weight percent of the pharmaceutical compositions or dosage forms into which they are incorporated.

[0272] GO-HA Formulation(s) (Including Compound(s) of Formula (I) and Any Embodiments Thereof) In one or more embodiments, a pharmaceutical composition (e.g., for treating a wound) is provided that includes a matrix component comprising a conjugate of graphene oxide (GO) and hyaluronic acid (HA) (wherein the GO and HA are covalently bonded via a linker); a compound of Formula (I) (or any embodiment thereof, including Compound 1, Compound 7, or Compound 8 in some embodiments); and water. The covalently bonded GO and HA are also referred to herein as GO-HA conjugates or simply GO-HA. GO-HA conjugates may be prepared according to procedures known to those skilled in the art, including those disclosed in US-2019-0105398-A1.

[0273] Graphene oxide (GO), as used herein, refers to the oxidized form of graphene, which is a single-layer form of graphite. GO can be obtained by treating graphite with a strong oxidizing agent. GO contains varying amounts of carbon, oxygen, and hydrogen depending on its production method. Its planar length can range from several hundred nanometers to several micrometers, and its thickness can be approximately 0.7-1.2 nm. When produced using sulfuric acid (e.g., the Hummers method), GO can contain various oxygen-containing moieties, such as oxygen epoxide groups, carboxylic acids (-COOH), and phenols. An example structure of GO is shown below.

[0274] [ka]

[0275] Hyaluronic acid (HA) is an anionic, highly hydrophilic, non-sulfated glycosaminoglycan that occurs naturally throughout the human body. It can be several thousand carbohydrate units long and can bind water to give a firm, viscous gel. An example structure of HA is shown below:

[0276] [ka]

[0277] In the compositions of the present disclosure, GO and HA are covalently linked to form a matrix component (or carrier) that serves to form a stable suspension of the compound of Formula (I) (e.g., Compound 1, Compound 7, or Compound 8), and may provide other concomitant benefits to wound healing. The covalent linkage may be achieved through the use of a linker or linker moiety ("GO-HA linker"). In one or more embodiments, the GO-HA linker may contain 2-25 carbons. In one or more embodiments, the GO-HA linker is linear. In one or more embodiments, the GO-HA linker is branched. The GO-HA linker may be saturated or unsaturated.

[0278] In one or more embodiments, the GO-HA linker is C2-C 25 It may include alkylene groups, where the carbon and hydrogen in the alkylene group may be replaced by oxygen or other atoms or groups such as hydroxy, carboxy, amino, alkyl, alkoxy, alkenyl, alkynyl, nitro, etc. In one or more embodiments, the GO-HA linker may include one or more CH2CH2O- units.

[0279] In one or more embodiments, the GO-HA linker is -R x -R s -R y -(In the formula, R x and R y are each independently selected from the group consisting of -CO-, -COO-, -H-, -HH-, -HH-CO-, -CS-, -S-, and -O-; and R s (also referred to herein as a spacer group) may be an unsubstituted or substituted, saturated or unsaturated, straight chain alkylene group having 2 to 20 backbone carbons. In one or more embodiments, R x and R y are *-HH-CO- (* indicates the end of the linker distal to Rs).

[0280] In one or more embodiments, the spacer group in the GO-HA linker can be an unsubstituted or substituted, saturated or unsaturated, linear alkylene group having 2 to 20 backbone carbons. By way of example and not limitation, HA can be substituted with the following spacer groups:

[0281] [ka]

[0282] (In the formula, R 101 and R 102may independently be -CONHNH-, -S-, -NH-, -O- or other nucleophiles, and n is an integer and can be, for example, 1 to 20, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc. In one or more embodiments, the HA is derivatized with a spacer group comprising a dihydrazide (e.g., -NHNHC(O)-alkylene-CONHNH-), such as adipic acid dihydrazide (-NHNHC(O)(CH2)4CONHNH-).

[0283] In one or more embodiments of the GO-HA pharmaceutical composition, the weight ratio of the compound of Formula (I) (e.g., Compound 1, Compound 7, or Compound 8) to GO-HA can be about 1:100 to 100:1, for example, about 1:2 to about 2:1. In one or more embodiments of the GO-HA conjugate, the weight ratio of GO:HA can be about 1:1 to about 1:20, or about 1:6 to about 1:10.

[0284] In one or more embodiments, the GO-HA pharmaceutical composition further comprises a pharmaceutical carrier (e.g., an excipient), compound, or material that allows the composition to be presented in the form of a semi-solid aqueous gel that can be topically administered. For example, carboxymethylcellulose may be used as a gel-forming agent. However, other cellulose derivatives such as crystalline cellulose, as well as polysaccharides such as alginate, agarose, tragacanth, guar gum, and xantham gum, are also suitable gel-forming agents. The gel may be made thicker and / or firmer, if necessary, by adding a relatively elastic gel-forming material such as a cross-linked fibrous protein, e.g., gelatin or collagen cross-linked with formaldehyde. In one or more embodiments, the GO-HA pharmaceutical composition may be in the form of a cream, which may include excipients suitable for cream formulations, such as paraffin oil, petrolatum, wax, organic esters such as cetyl palmitate, and the like.

[0285] In one or more embodiments, the GO-HA pharmaceutical composition of the present disclosure further comprises a thickener to achieve a desired viscosity of the composition for dermal delivery. For example, the thickener may include hydroxypropyl cellulose (HPC). HPC can form the GO-HA pharmaceutical composition into a smooth film that is easy to apply. It can also reduce evaporation, keeping wounds moist longer, a factor that has been shown to improve healing and reduce scarring. Different grades of UPC are available, depending on the molecular weight or viscosity of a given concentration of aqueous UPC solution.

[0286] In one or more embodiments of the GO-HA pharmaceutical composition, the compound of Formula (I) (e.g., Compound 1, Compound 7, or Compound 8) may comprise about 0.001 wt% to about 5 wt% of the total composition (including water). In one or more embodiments of the GO-HA pharmaceutical composition, the compound of Formula (I) (e.g., Compound 1, Compound 7, or Compound 8) may comprise about 0.01 wt% to about 2 wt%, about 0.02 wt% to about 1 wt%, or about 0.05 wt% to about 0.5 wt% of the total composition. In one or more embodiments, GO-HA comprises about 0.001 wt% to about 5 wt% of the total composition. In one or more embodiments, GO-HA may comprise from about 0.01 wt% to about 2 wt%, from about 0.02 wt% to about 1 wt%, or from about 0.05 wt% to about 0.5 wt% of the total composition.

[0287] Generally, the GO-HA pharmaceutical composition may appear as a slightly dark or black viscous liquid overall. The compound of Formula (I) (e.g., Compound 1, Compound 7, or Compound 8) is uniformly dispersed in the viscous suspension, which is stable at room temperature for several months. In one or more embodiments, the composition further comprises a surfactant that enhances the miscibility or solubility of hydrophobic substances in water. In one or more embodiments, the surfactant may be a non-ionic hydrophilic substance such as polyethylene glycol (PEG). The PEG can have a number average molecular weight of about 100 to about 10,000 daltons, or about 200 to about 4000 daltons, e.g., about 200 to about 1000, about 200 to about 800, about 200 to about 500, about 200 to about 400, about 300 to about 400, about 350 to about 450, about 200, about 250, about 300, about 350, about 400, about 450, about 500, about 550, about 600, about 650, about 700, about 750, about 800, about 850, about 900, about 950, about 1000 daltons, etc. In one or more embodiments, the PEG can be present in the composition in an amount of about 0.1 to about 20 wt% of the total composition. For example, PEG can be about 0.2 wt% to about 10 wt%, or about 0.5 wt% to about 10 wt%, or about 1 wt% to about 10 wt% of the total composition. Other non-ionic hydrophilic materials, such as copolymers of PEG and PPG (polypropylene glycol), such as poloxamers, can also be used. In one example, Poloxamer-188 (having an average molecular weight of about 8400 daltons) can be used.

[0288] The GO-HA pharmaceutical compositions described herein can include other medicinal or therapeutic compounds in addition to a compound of Formula (I) (e.g., Compound 1, Compound 7, or Compound 8). In other words, a GO-HA pharmaceutical composition in which a compound(s) of Formula (I) (e.g., Compound 1, Compound 7, or Compound 8) is present can also function as a base dispersion medium into which other medicinal or therapeutic agents (particularly those that are hydrophobic) can be dispersed for topical administration, for example, to a wound. These agents can include anti-fibrotic compounds (e.g., pirfenidone, halofuginone, nintedanib, tocilizumab, rilonacept, etc.), anti-cancer agents, anti-inflammatory agents, analgesics, antibiotics, Wnt inhibitors, hedgehog pathway inhibitors, TGF-β inhibitors, LOX inhibitors, etc.

[0289] Sustained-release dosage forms Active ingredients, such as the compounds provided herein, may be administered by controlled release means or by delivery devices known to those skilled in the art. In some or all embodiments, U.S. Patent Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123; 4,008,719; 5,674,533; 5,059,595; 5,591,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; 5,639,480; 5,733,566; 5,739,108; 5,891,474; 5,922,356; 5,972, 891; 5,980,945; 5,993,855; 6,045,830; 6,087,324; 6,113,943; 6,197,350; 6,248,363; 6,264,970; 6,267,981; 6,376,461; 6,419,961; 6,589,548; 6,613,358; and 6,699,500 (each of which is incorporated by reference herein in its entirety). In some or all embodiments, such dosage forms may use hydropropylmethylcellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres, or combinations thereof to provide sustained or controlled release of one or more active ingredients, and may be used to provide desired release profiles at various rates.Suitable controlled-release formulations known to those skilled in the art, including those described herein, can be easily selected for use with the active ingredients provided herein.Therefore, unit dosage forms suitable for oral administration, such as, but not limited to, tablets, capsules, gel caps, and caplets, adapted for controlled release, are encompassed herein.

[0290] All controlled-release formulations share a common goal of improving drug therapy over that achieved by their non-controlled counterparts. Ideally, the use of an optimally designed controlled-release formulation in a medical treatment is characterized by the use of a minimum amount of drug substance to cure or control a condition in a minimum amount of time. Advantages of controlled-release formulations include extended drug activity, reduced dosing frequency, and improved compliance. Additionally, controlled-release formulations can be used to affect the time of onset of action or other characteristics, such as blood levels of the drug, which can then affect the occurrence of side effects (e.g., adverse effects).

[0291] Most controlled-release formulations are designed to initially release an amount of drug (active ingredient) that quickly produces the desired therapeutic effect, and then gradually and continuously release another amount of drug to maintain this level of therapeutic or prophylactic effect over an extended period of time. To maintain this constant level of drug in the body, the drug must be released from the dosage form at a rate that will replace the amount of drug being metabolized and excreted from the body. Controlled-release of an active ingredient can be stimulated by various conditions, including, but not limited to, pH, temperature, enzymes, water, or other physiological conditions or compounds.

[0292] In some or all embodiments, the drug may be administered using intravenous infusion, an implantable osmotic pump, a transdermal patch, liposomes, or other modes of administration. In some or all embodiments, a pump may be used (see Sefton, CRC Crit. Ref. Biomed. Eng. 14:201 (1987); Buchwald et al., Surgery 88:507 (1980); Saudek et al., N. Engl. J. Med. 321:574 (1989)). In another embodiment, a polymeric material may be used. In yet another embodiment, a controlled release system may be placed in a mammal at an appropriate site determined by one skilled in the art, i.e., thus requiring only a fraction of the systemic dose (see, e.g., Goodson, Medical Applications of Controlled Release, vol. 2, pp. 115-138 (1984)). Other controlled release systems are discussed in the review by Langer (Science 249:1527-1533 (1990)).An outer polymeric membrane, such as polyethylene, polypropylene, ethylene / propylene copolymer, ethylene / ethyl acrylate copolymer, ethylene / vinyl acetate copolymer, silicone rubber, polydimethylsiloxane, neoprene rubber, chlorinated polyethylene, polyvinyl chloride, vinyl acetate, vinylidene chloride, vinyl chloride copolymers with ethylene and propylene, ionomer polyethylene terephthalate, butyl rubber epichlorohydrin rubber, ethylene / vinyl alcohol copolymer, ethylene / vinyl acetate / vinyl alcohol terpolymer, and ethylene / vinyloxyethanol copolymer, which are insoluble in body fluids. The active ingredient may be dispersed in a solid inner matrix surrounded by a conjugate, such as polymethyl methacrylate, polybutyl methacrylate, plasticized or unplasticized polyvinyl chloride, plasticized nylon, plasticized polyethylene terephthalate, natural rubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene, ethylene-vinyl acetate copolymer, silicone rubber, polydimethylsiloxane, silicone carbonate copolymer, hydrophilic polymers such as hydrogels of acrylic and methacrylic acid esters, collagen, cross-linked polyvinyl alcohol, and cross-linked partially hydrolyzed polyvinyl acetate. The active ingredient then diffuses through the outer polymeric membrane in a release rate-controlling step. The percentage of active ingredient in such parenteral compositions largely depends on the specific nature of the composition and the needs of the mammal.

[0293] Parenteral dosage forms In some or all embodiments, parenteral dosage forms are provided. Parenteral dosage forms can be administered to mammals by various routes, including, but not limited to, subcutaneous, intravenous (including bolus injection), intramuscular, and intraarterial. Because their administration typically bypasses the mammal's natural defenses against contaminants, parenteral dosage forms are typically sterile or capable of being sterilized before administration to a mammal. In some or all embodiments, parenteral dosage forms include, but are not limited to, ready-to-inject solutions, dry products ready to be dissolved or suspended in a pharmaceutically acceptable vehicle for injection, ready-to-inject suspensions, and emulsions.

[0294] Suitable vehicles that can be used to provide parenteral dosage forms are well known to those skilled in the art. In some or all embodiments, suitable vehicles include, but are not limited to, water for injection USP; aqueous vehicles such as, but not limited to, sodium chloride injection, Ringer's injection, dextrose injection, dextrose and sodium chloride injection, and lactated Ringer's injection; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol; non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

[0295] Compounds that increase the solubility of one or more of the active ingredients disclosed herein can also be incorporated into the parenteral dosage forms.

[0296] Transdermal, topical and mucosal dosage forms Transdermal, topical, and mucosal dosage forms are also provided. Transdermal, topical, and mucosal dosage forms include, but are not limited to, eye drops, sprays, aerosols, creams, lotions, ointments, gels, solutions, emulsions, suspensions, or other dosage forms known to those skilled in the art. See, for example, Remington: The Science and Practice of Pharmacy; Pharmaceutical Press; 22nd Edition (September 15, 2012); and Introduction to Pharmaceutical Dosage Forms, 4th Edition, Lea & Febiger, Philadelphia (1985). Dosage forms suitable for treating mucosal tissues in the oral cavity may be formulated as mouthwashes or oral gels. Further, transdermal dosage forms include "reservoir-type" or "matrix-type" patches, which are applied to the skin and worn for a specific period of time to allow penetration of a desired amount of active ingredient.

[0297] Suitable carriers (e.g., excipients and diluents) and other materials that can be used to provide the transdermal, topical, and mucosal dosage forms encompassed herein are well known to those skilled in the art of pharmacy and depend on the specific tissue to which a given pharmaceutical composition or dosage form is to be applied. With this in mind, typical excipients include, but are not limited to, water, acetone, ethanol, ethylene glycol, propylene glycol, butane 1,3 diol, isopropyl myristate, isopropyl palmitate, mineral oil, and mixtures thereof to form non-toxic, pharmaceutically acceptable lotions, tinctures, creams, emulsions, gels, or ointments. Moisturizers or humectants may also be added to pharmaceutical compositions and dosage forms as desired. Examples of such additional ingredients are well known in the art. See, for example, Remington: The Science and Practice of Pharmacy; Pharmaceutical Press; 22nd Edition (September 15, 2012).

[0298] Depending on the specific tissue to be treated, additional ingredients may be used before, in conjunction with, or after treatment with the provided active ingredient. In some or all embodiments, a penetration enhancer may be used to assist in delivering the active ingredient to the tissue. Suitable penetration enhancers include, but are not limited to: acetone; various alcohols such as ethanol, oleyl, and tetrahydrofuryl; alkyl sulfoxides such as dimethyl sulfoxide; dimethylacetamide; dimethylformamide; polyethylene glycol; pyrrolidones such as polyvinylpyrrolidone; Kollidon grade (povidone, polyvidone); urea; and various water-soluble or insoluble sugar esters such as Tween 80 (polysorbate 80) and Span 60 (sorbitan monostearate).

[0299] The pH of pharmaceutical compositions or dosage forms, or the pH of the tissue to which pharmaceutical compositions or dosage forms are applied, can also be adjusted to improve the delivery of one or more active ingredients.Similarly, the polarity of solvent carriers, their ionic strength, or tonicity can also be adjusted to improve delivery.Compounds such as stearates can also be added to pharmaceutical compositions or dosage forms to advantageously change the hydrophilicity or lipophilicity of one or more active ingredients, thereby improving delivery.In this regard, stearates can function as lipid vehicles for formulation, as emulsifiers or surfactants, and as delivery-enhancing or penetration-enhancing agents.Different salts, hydrates, or solvates of active ingredients can be used to further adjust the properties of the resulting composition.

[0300] Dosage and unit dosage form In treating humans, a physician will determine the posology that he or she deems most appropriate depending on the treatment required (e.g., preventative and curative), as well as the age, weight, stage of the disease, disorder, or condition, and other factors specific to the mammal being treated. In some or all embodiments, a topical dose is administered within 1 cm of the treatment site (e.g., wound). 2 It is listed as mg per cm, and ranges from about 0.001 to about 50 mg / cm 2 , or about 0.005 to about 50 mg / cm 2 , or about 0.01 to about 50 mg / cm 2 , or about 0.01 to about 40 mg / cm 2 , or about 0.01 to about 30 mg / cm 2 , or about 0.01 to about 20 mg / cm 2 , or about 0.01 to about 10 mg / cm 2 , or about 0.05 to about 10 mg / cm 2 , or about 0.05 to about 1 mg / cm 2 is.

[0301] In some or all embodiments, the topical and non-topical doses are about 1 to about 1000 mg per day for an adult, or about 5 to about 250 mg per day, or about 10 to about 50 mg per day for an adult. In some or all embodiments, the dose is about 5 to about 400 mg per day, or about 25 to about 200 mg per day per adult. In some or all embodiments, dose rates of about 50 to about 500 mg per day are also contemplated. In some or all embodiments, the subcutaneous dose is about 1 to about 50 mg per day, or about 1 to about 25 mg per day, or about 1 to about 10 mg per day, or about 1 to about 20 mg per day, or about 5 to about 25 mg per day, or about 5 mg to about 20 mg per day, or about 10 to about 20 mg per day. In some or all embodiments, the oral administration dose is about 0.01 mg to about 100 mg per day, about 0.01 to about 100 mg per day, or about 0.01 mg to about 50 mg per day, about 0.01 to about 25 mg per day, about 0.01 to about 15 mg per day, about 0.01 to about 10 mg per day, about 0.05 to about 1 ...25 mg per day, about 0.01 to about 15 mg per day, about 0.01 to about 25 mg per day The daily dose may be about 0.05 to about 5 mg per day, about 0.05 to about 1 mg per day, about 0.1 to about 100 mg per day, about 0.1 to about 50 mg per day, about 0.1 to about 25 mg per day, about 0.1 to about 15 mg per day, about 0.1 to about 10 mg per day, about 0.1 to about 5 mg per day, or about 0.5 mg to about 1 mg per day, or about 10 mg to about 200 mg per day. In some or all embodiments, including any of the above-mentioned embodiments, the daily dose may be administered once per day. In some or all embodiments, including any of the above-mentioned embodiments, the daily dose may be divided and administered twice per day. In some or all embodiments, including any of the above-mentioned embodiments, the daily dose may be divided and administered three times per day.

[0302] In some embodiments, the mg / day amount is for an adult. In a further aspect, provided is a method for treating a disease, disorder, or condition associated with Wnt signaling pathway activity in a mammal by administering to a mammal in need thereof a therapeutically or prophylactically effective amount of a compound provided herein, or a pharmaceutically acceptable salt thereof. The amount of a compound or composition that is therapeutically or prophylactically effective in treating a disease or one or more symptoms thereof will vary depending on the nature and severity of the disease or condition, and the route by which the active ingredient is administered. The frequency and dosage will also vary according to factors specific to each mammal, depending on the particular therapy (e.g., therapeutic or prophylactic agent) administered, the severity of the disorder, disease, or condition, the route of administration, and the mammal's age, physical condition, weight, response, and past medical history. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0303] Exemplary dosages of the compositions in some or all embodiments include mg or μg amounts of active compound per kg of mammal or sample weight (e.g., from about 10 μg / kg to about 50 mg / kg, from about 100 μg / kg to about 25 mg / kg, or from about 100 μg / kg to about 10 mg / kg). For the compositions provided herein, the dosage administered to a mammal in some or all embodiments is from 0.01 mg / kg to 3 mg / kg of the mammal's body weight, or from 0.10 mg / kg to 3 mg / kg of the mammal's body weight, based on the weight of the active compound. In some or all embodiments, the dosage administered to a mammal is between 0.20 mg / kg and 2.00 mg / kg, or between 0.30 mg / kg and 1.50 mg / kg of the mammal's body weight. In some embodiments, the dosage is administered subcutaneously to the mammal and is between about 0.01 mg / kg and 1 mg / kg (inclusive), or between about 0.03 mg / kg and 0.5 mg / kg (inclusive), of the mammal's body weight, based on the weight of the active compound. In some embodiments, the dosage is administered orally to the mammal and is between about 0.10 mg / kg and 5 mg / kg (inclusive), or between about 0.10 mg / kg and 2 mg / kg (inclusive), of the mammal's body weight, based on the weight of the active compound. In some or all embodiments, the recommended daily topical dosage range of the compositions provided herein for the conditions described herein is in the range of about 0.01 mg to about 100 mg per day, given as a single dose once daily or as divided doses (e.g., two or three doses) throughout the day.

[0304] In some or all embodiments, the recommended daily dose range of the compositions provided herein for the conditions described herein is in the range of about 0.1 mg to about 1000 mg per day, given as a single dose once daily or as divided doses throughout the day. In some or all embodiments, the daily dose is administered twice daily in equally divided doses. In some or all embodiments, the daily dose is administered three times daily in equally divided doses. In some or all embodiments, the daily dose range should be about 0.01 mg to about 400 mg per day, about 0.1 mg to about 250 mg per day, about 10 mg to about 200 mg per day, or in further embodiments, about 10 mg to about 150 mg per day, between about 25 mg and about 100 mg per day. As will be apparent to one of skill in the art, in some cases it may be necessary to use doses of the active ingredients outside the ranges disclosed herein. Furthermore, it should be noted that the clinical or treating physician will know how and when to interrupt, adjust, or terminate therapy in conjunction with the mammal's response.

[0305] Different therapeutically effective amounts may be applicable to different diseases and conditions, as will be readily known to those skilled in the art. Similarly, amounts sufficient to prevent, manage, treat, or ameliorate such disorders, but insufficient to cause or reduce side effects associated with the compositions provided herein, are also encompassed by the dosage and dose frequency schedules described herein. Furthermore, when a mammal is administered multiple doses of the compositions provided herein, not all doses need to be the same. In some or all embodiments, the dosage administered to a mammal may be increased to improve the prophylactic or therapeutic effect of the composition, or it may be decreased to reduce one or more side effects experienced by a particular mammal.

[0306] In some or all embodiments, the dosage of the composition provided herein, based on weight of active compound, administered to prevent, treat, manage, or ameliorate a disorder or one or more symptoms thereof in a mammal is about 0.01 mg / kg, about 0.1 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 10 mg / kg, or about 15 mg / kg or more of the mammal's body weight. In another embodiment, the dosage of the composition or a composition provided herein, administered to prevent, treat, manage, or ameliorate a disorder or one or more symptoms thereof in a mammal, is about 0.01 mg / kg, about 0.1 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 10 mg / kg, or about 15 mg / kg or more of the mammal's body weight. The dosage herein) is selected from about 0.01 mg / kg to about 100 mg / kg, selected from about 0.1 mg to about 200 mg, selected from about 0.1 mg to about 100 mg, selected from about 0.1 mg to about 50 mg, selected from about 0.1 mg to about 25 mg, selected from about 0.1 mg to about 20 mg, selected from about 0.1 mg to about 15 mg, selected from about 0.1 mg to about 10 mg, selected from about 0.1 mg to about 7.5 mg, selected from about 0.1 mg to about 5 mg, selected from 0.1 to about 2.5 mg, selected from about 0.25 mg to about 20 mg. The unit dose is selected from about 0.25 to about 15 mg, selected from about 0.25 mg to 12 mg, selected from about 0.25 to about 10 mg, selected from about 0.25 mg to about 7.5 mg, selected from about 0.25 mg to about 5 mg, selected from about 0.5 mg to about 2.5 mg, selected from 1 mg to about 20 mg, selected from about 1 mg to about 15 mg, selected from about 1 mg to about 12 mg, selected from 1 mg to about 10 mg, selected from about 1 mg to about 7.5 mg, selected from about 1 mg to about 5 mg, or selected from about 1 mg to about 2.5 mg.

[0307] In some or all embodiments, a dose of a compound or composition provided herein can be administered to achieve a steady-state concentration of the active ingredient in the blood or serum of a mammal. The steady-state concentration can be determined by measurement according to techniques available to those skilled in the art, or can be determined based on the mammal's physical characteristics, such as height, weight, and age. In some or all embodiments, repeated administrations of the same composition can be administered, and the administrations can be separated by at least 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75 days, 3 months, or 6 months. In other embodiments, repeated administrations of the same prophylactic or therapeutic agent can be administered, and the administrations can be separated by at least 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75 days, 3 months, or 6 months.

[0308] In some or all embodiments, provided herein are unit dosages comprising the compound or a pharmaceutically acceptable salt thereof in a form suitable for administration. Such forms are described in detail herein. In some or all embodiments, the unit dosage contains 1-1000 mg, 1-100 mg, or 10-50 mg of active ingredient. In certain embodiments, the unit dosage contains about 1, 5, 10, 25, 50, 100, 125, 250, 500, or 1000 mg of active ingredient. Such unit dosages may be prepared according to techniques well known to those skilled in the art.

[0309] In some or all embodiments, dosages of the second agent used in the combination therapy are provided herein. In some or all embodiments, lower dosages than those that have been or are currently used to treat the disease, disorder, or condition are used in the combination therapy provided herein. Recommended dosages of the second agent can be obtained from the knowledge of those skilled in the art. For second agents approved for clinical use, recommended dosages are found, for example, in Hardman et al. (eds.), 1996, Goodman & Gilman's The Pharmacological Basis of Therapeutics, 9th Edition, McGraw-Hill, New York; Physician's Desk Reference (PDR), 57th Edition, 2003, Medical Economics Co., Inc., Montvale, NJ (which are incorporated herein by reference in their entireties).

[0310] In various embodiments, the therapies (e.g., a compound provided herein and a second agent) are administered less than 5 minutes apart, less than 30 minutes apart, 1 hour apart, about 1 hour apart, about 1-2 hours apart, about 2 to about 3 hours apart, about 3 to about 4 hours apart, about 4 to about 5 hours apart, about 5 to about 6 hours apart, about 6 to about 7 hours apart, about 7 to about 8 hours apart, about 8 to about 9 hours apart, about 9 to about 10 hours apart, about 10 to about 11 hours apart, about 11 to about 12 hours apart, about 12 to 18 hours apart, 18 to 24 hours apart, 24 to 36 hours apart, 36 to 48 hours apart, 48 to 52 hours apart, 52 to 60 hours apart, 60 to 72 hours apart, 72 to 84 hours apart, 84 to 96 hours apart, or 96 to 120 hours apart. In various embodiments, the therapies are administered within 24 hours or within 48 hours. In some or all embodiments, two or more therapies are administered within the same patient visit. In other embodiments, a compound provided herein and a second agent are administered simultaneously.

[0311] In other embodiments, the compound provided herein and the second agent are administered about 2-3 days apart, 2-4 days apart, about 4-6 days apart, about 1 week apart, about 1-2 weeks apart, or more than 2 weeks apart.

[0312] In some or all embodiments, the administration of the same agent can be repeated and the administrations can be separated by at least 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75 days, 3 months, or 6 months. In other embodiments, the administration of the same agent can be repeated and the administrations can be separated by at least 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75 days, 3 months, or 6 months.

[0313] In some or all embodiments, a compound provided herein and a second active agent are administered to a patient, a mammal such as a human in some or all embodiments, in an order and within a time interval such that the compound provided herein can act together with the other agent to provide an enhanced benefit over the other agent when administered in other ways. In some or all embodiments, the second active agent can be administered simultaneously or sequentially at different times in any order, but if not administered simultaneously, they should be administered sufficiently close in time to provide the desired therapeutic or prophylactic effect. In some or all embodiments, the compound provided herein and the second active agent exert their effects at overlapping times. Each second active agent can be administered separately in any suitable form and by any suitable route. In other embodiments, the compound provided herein is administered before, simultaneously with, or after the administration of the second active agent.

[0314] In some or all embodiments, a compound provided herein and a second agent are cyclically administered to a patient. Cycling therapy involves administering a first agent (e.g., a first prophylactic or therapeutic agent) for a period of time, followed by administration of a second agent and / or a third agent (e.g., a second and / or third prophylactic or therapeutic agent) for a period of time, and repeating this sequential administration. Cycling therapy may prevent the development of resistance to one or more therapies, avoid or reduce side effects of one therapy, and / or improve the efficacy of the treatments.

[0315] In some or all embodiments, the compound provided herein and the second active agent are administered in cycles of less than about 3 weeks, about once every 2 weeks, about once every 10 days, or about once a week. One cycle can include administering the compound provided herein and the second agent by infusion over about 90 minutes per cycle, about 1 hour per cycle, or about 45 minutes per cycle. Each cycle can include at least one week of rest, at least two weeks of rest, or at least three weeks of rest. The number of cycles administered is about 1 to about 12 cycles, more typically about 2 to about 10 cycles, and more typically about 2 to about 8 cycles.

[0316] In other embodiments, the treatment courses are administered to the patient simultaneously, i.e., individual doses of the second agent are administered separately but within a time interval that allows the compounds provided herein to act in concert with the second active agent. In some or all embodiments, one component may be administered once a week, with the other component being administered once every two weeks or once every three weeks. In other words, the dosing regimen is simultaneous even if the therapeutic agents are not administered at the same time or within the same day.

[0317] The second agent may act additively or synergistically with the compound provided herein. In some or all embodiments, the compound provided herein is administered simultaneously with one or more second agents in the same pharmaceutical composition. In another embodiment, the compound provided herein is administered simultaneously with one or more second agents in separate pharmaceutical compositions. In yet another embodiment, the compound provided herein is administered before or after administration of the second agent. Also contemplated are administration of the compound provided herein and the second agent by the same or different routes of administration, e.g., oral and parenteral. In some or all embodiments, when the compound provided herein is administered simultaneously with a second agent that may produce adverse side effects, including but not limited to toxicity, the second active agent may advantageously be administered at a dose below the threshold at which adverse side effects occur.

[0318] kit Kits for use in methods of treating diseases, disorders, or conditions associated with Wnt signaling pathway activity are also provided. The kits may include a compound or composition provided herein, a second agent or composition, and instructions providing a healthcare provider with information regarding use to treat a disease, disorder, or condition associated with Wnt signaling pathway activity. The instructions may be provided in printed form, or in the form of an electronic medium such as a floppy disk, CD, or DVD, or in the form of a website address where such instructions can be obtained. A unit dose of a compound or composition provided herein, or a second agent or composition, may comprise a dose that, when administered to a mammal, can maintain a therapeutically or prophylactically effective plasma level of the compound or composition in the mammal for at least one day. In some or all embodiments, the compound or composition may be included as a sterile aqueous pharmaceutical composition or a dry powder (e.g., lyophilized) composition.

[0319] In some embodiments, suitable packaging is provided. As used herein, "packaging" includes solid matrices or materials conventionally used in systems and capable of retaining, within certain limits, the compounds provided herein and / or a second agent suitable for administration to a mammal. Such materials include glass and plastic (e.g., polyethylene, polypropylene, and polycarbonate) bottles, vials, paper, plastic, and plastic-foil laminate envelopes, and the like. When electron beam sterilization techniques are employed, the packaging should have a sufficiently low density to allow sterilization of the contents.

[0320] How to use Provided herein is a method for inhibiting the Wnt transcriptional signaling pathway in a mammal, comprising contacting administration of an effective amount of a compound of formula (I) (including a single stereoisomer or a mixture of stereoisomers thereof; and / or a pharmaceutically acceptable salt thereof).

[0321] Provided herein is a method for treating a disease, disorder, or condition associated with Wnt transcription or Wnt signaling pathway activity in a mammal, comprising administering a therapeutically or prophylactically effective amount of a compound of Formula (I) described herein or a pharmaceutical composition described herein. In one or more embodiments, the method is for treating a disease, disorder, or condition associated with Wnt transcription or Wnt signaling pathway activity and comprises administering a compound of Formula (I) (or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition according to one or more embodiments to a mammal in need thereof.

[0322] In one or more embodiments, the method is for stimulating tissue regeneration at a wound in a mammal in need thereof. In one or more embodiments, the method comprises contacting the wound with an effective amount of a compound of Formula (I) (or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition according to one or more embodiments. In some embodiments, the mammal is a human. In one group of embodiments, the disease, disorder, or condition is a wound.

[0323] In one group of embodiments, the disease, disorder, or condition is selected from chronic wounds, acute wounds, alkali-burned corneal wounds, burns, lesions (including lesions caused by viruses selected from HPV and / or the Poxviridae family of viruses), inflammatory skin diseases (including acne, psoriasis, rosacea, and scleroderma), cartilage diseases (including osteoarthritis, rheumatoid arthritis, internal joint derangements, and degenerative cartilage diseases), bone diseases (including osteoporosis), organ fibrosis (including pulmonary fibrosis, cardiac fibrosis, liver fibrosis, and kidney fibrosis), cancer (including melanoma, breast cancer, and prostate cancer), denervated body parts in need of reinnervation, tissues in need of regeneration (including damaged elastic cartilage), bacterial growth in need of inhibition, fungal growth in need of inhibition, tissues in need of angiogenesis, osteoclast differentiation in need of inhibition, impaired osteoblast differentiation (where inhibition of osteoblast differentiation is required), and / or bone destruction associated with breast cancer.

[0324] In some or all embodiments, provided herein are methods for treating a disease, disorder, or condition associated with Wnt transcription or Wnt signaling pathway activity in a mammal in need thereof. In some or all embodiments, the method comprises administering to a subject in need thereof a therapeutically or prophylactically effective amount of a compound effective to treat a disease, disorder, or condition associated with Wnt transcription or Wnt signaling pathway activity in combination with a second agent effective to treat the condition associated with Wnt transcription or Wnt signaling pathway activity. The compound can be any compound described herein, and the second agent can be any second agent described in the art or herein. In some or all embodiments, the compound is in the form of a pharmaceutical composition or dosage form, as described elsewhere herein.

[0325] In some or all embodiments, provided herein are methods of inhibiting Wnt transcription or Wnt signaling pathway activity, comprising contacting Wnt with a compound of formula (I) or a compound selected from compounds 1-44.

[0326] In one or more embodiments, the present disclosure provides a method for improving wound healing, comprising contacting a wound with an effective amount of a composition of the present disclosure. Injured wounds are contemplated to include, but are not limited to, those resulting from surgical wounds (such as lacerations, abrasions, cuts, scrapes, or punctures caused by knives, scalpels, bullets, or other sharp or blunt objects) caused by physical impact that disrupts the structure and function of the skin. The present disclosure is contemplated for use on wounds caused by excessive (low or high) temperatures, such as burns, ionizing radiation, chemotherapy, or unexpected acute injuries resulting from accidents or mishaps. The present disclosure is contemplated for use on chronic wounds resulting from underlying conditions, such as diabetic ulcers.

[0327] The composition(s) of the present disclosure described herein may be administered by topically applying the composition(s) to the wound. When the composition is contained in a medical device described herein that includes a substrate such as a patch or pad, the medical device may be secured to the wound so that the composition contacts the wound.

[0328] In the methods of making the compositions used in the methods of the present disclosure, the spacer group can be an unsubstituted or substituted, saturated or unsaturated, linear alkylene group having 2 to 20 backbone carbons. By way of example and not limitation, reagents for derivatizing HA include the following:

[0329] [ka]

[0330] (In the formula, R 101 and R 102 can independently be -CONHNH, -SH, -NH, -OH, or other nucleophiles, and n is an integer and can be, for example, 1 to 20, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc. In one or more embodiments, the reagent for derivatizing HA can be a dihydrazide (e.g., -NHNHC(O)-alkylene-CONHNH2), such as adipic acid dihydrazide (e.g., -NHNHC(O)(CH2)4CONHNH2).

[0331] In one or more embodiments, a method of making a composition of the present disclosure includes obtaining GO-HA (e.g., by a method herein), adding or dissolving the GO-HA conjugate in water to obtain an aqueous GO-HA solution, and adding a compound of formula (I) (or any embodiment thereof, in some embodiments including compound 1, compound 7, or compound 8) to the aqueous GO-HA solution to form a mixture (GO-HA + a compound of formula (I) or any embodiment thereof, in some embodiments including GO-HA + compound 1, GO-HA + compound 7, or GO-HA + compound 8). In some instances, this is accomplished by first adding or dissolving a compound of formula (I) (or any embodiment thereof, in some embodiments including Compound 1, Compound 7, or Compound 8) in a non-ionic hydrophilic polymer, such as PEG-400 (or PEG400 having an average molar mass of about 400), and then adding the solution of a compound of formula (I) (or any embodiment thereof, in some embodiments including Compound 1, Compound 7, or Compound 8) to an aqueous solution of GO-HA conjugate to produce GO-HA + a compound of formula (I) (or any embodiment thereof, in some embodiments including GO-HA + Compound 1, GO-HA + Compound 7, or GO-HA / Compound 8).

[0332] In some or all embodiments, the wound to be improved, treated, repaired, or healed is selected from one or more of the group consisting of acute wounds, chronic wounds, tear wounds, abrasions, laceration wounds, puncture wounds, avulsion wounds, skin incisions, surgical wounds, thermal wounds, burn wounds, ulcers, chemical wounds, bite wounds, stab wounds, gunshot wounds, other penetrating high velocity projectile wounds, puncture wounds, electrical wounds, amputation wounds, crush wounds, poison wounds, radiation wounds, scalping wounds, penetrating wounds, incision wounds, blunt force trauma wounds, skin lacerations, internal wounds, open wounds, closed wounds, excision wounds, infected wounds, exuding wounds, non-healing wounds, wounds associated with dressing changes, amputations, necrotizing fasciitis wounds, osteomyelitis wounds, and post-traumatic wounds.

[0333] The present disclosure contemplates use in chronic wounds that arise as a result of an underlying condition, such as a diabetic ulcer.

[0334] In some or all embodiments, the wound to be improved, treated, repaired, or healed is a wound in an acute care setting, including post-surgery. In some or all embodiments, the wound to be improved, treated, repaired, or healed is a wound in an acute care setting, including post-surgery, and the compound is administered intravenously. In some or all embodiments, the wound to be improved, treated, repaired, or healed is a surgical wound. In some or all embodiments, the wound to be improved, treated, repaired, or healed is a surgical wound, and the compound is administered topically (e.g., as a spray). In some or all embodiments, the wound to be improved, treated, repaired, or healed is an acute or chronic wound. In some or all embodiments, the wound to be improved, treated, repaired, or healed is an acute or chronic wound, and the compound is administered subcutaneously. In some or all embodiments, the wound to be improved, treated, repaired, or healed is an acute or chronic wound, and the compound is administered orally.

[0335] In one group of embodiments, the wound to be ameliorated, treated, repaired, or healed is a burn. In one or more embodiments, the burn is a thermal burn. In one or more embodiments, the burn is a chemical burn. In one or more embodiments, the burn is an electrical burn. In one or more embodiments, the burn is a thermal burn. In one or more embodiments, the wound is a radiation burn. In one or more embodiments, the wound is a first-degree burn. In one or more embodiments, the wound is a second-degree burn. In one or more embodiments, the wound is a third-degree burn.

[0336] In some or all embodiments, the compounds described herein are used to delay the onset of wounds or reduce the severity or duration of wounds. In some or all embodiments, the compounds described herein are used to reduce the severity or duration of wounds associated with Wnt transcripts or Wnt signaling pathway activity. In some embodiments, the compounds described herein are used to delay or prevent the onset of wounds.

[0337] In some or all embodiments, the compounds described herein are used for the prevention of wounds or conditions associated with Wnt transcripts or Wnt signaling pathway activity.

[0338] In some or all embodiments, the compounds described herein are used for the treatment of wounds or conditions associated with Wnt transcripts or Wnt signaling pathway activity.

[0339] Assay Method Compounds may be assayed for effectiveness in treating a disease, disorder, or condition associated with Wnt signaling pathway activity according to any assay known to those skilled in the art. Exemplary assays are provided elsewhere herein.

[0340] Second treatment In some or all embodiments, the compounds and compositions provided herein are useful in methods of treating wounds and / or conditions associated with Wnt transcripts and / or Wnt signaling pathway activity, which methods further comprise administering a second agent effective in treating wounds and / or Wnt transcription-related disorders and / or conditions associated with Wnt transcripts and / or Wnt signaling pathway activity. The second agent used in the treatment methods can be any agent known to those skilled in the art to be effective in treating wounds and / or Wnt transcription-related disorders and / or conditions associated with Wnt transcripts and / or Wnt signaling pathway activity, including those currently approved by the U.S. Food and Drug Administration or other similar agencies in countries other than the United States. The second pharmaceutical agent (second agent) is one previously described herein and can be used in the treatment methods. In some or all embodiments, the second agent is a PARP inhibitor, an anti-inflammatory drug, or a Notch inhibitor.

[0341] In one or more embodiments, the second agent is one or more of a corticosteroid, a cytotoxic drug, an antibiotic, an antiseptic, nicotine, an antiplatelet drug, an NSAID, colchicine, an anticoagulant, a vasoconstrictor or immunosuppressant, a growth factor, an antibody, a protease, a protease inhibitor, an antimicrobial peptide, an adhesion peptide, a hemostatic agent, a live cell, honey, nitric oxide, an antifibrotic compound (such as pirfenidone, halofuginone, nintedanib, tocilizumab, rilonacept), an anticancer agent, an anti-inflammatory agent, an analgesic, a Wnt inhibitor, a Hedgehog pathway inhibitor, a TGF-β inhibitor, and / or an LOX inhibitor, etc.

[0342] In some or all embodiments, the compounds provided herein are administered in combination with one second agent. In further embodiments, the compounds provided herein are administered in combination with two second agents. In further embodiments, the compounds provided herein are administered in combination with two or more second agents.

[0343] As used herein, the term "in combination" includes the use of more than one therapies (e.g., one or more prophylactic and / or therapeutic agents). The use of the term "in combination" does not restrict the order in which therapies (e.g., prophylactic and / or therapeutic agents) are administered to a mammal with a disorder. A first therapy (e.g., a prophylactic or therapeutic agent such as a compound provided herein) can be administered to a mammal having a disorder prior to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), concomitantly with, or after (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) administration of a second therapy (e.g., a prophylactic or therapeutic agent).

[0344] As used herein, the term "synergistic" includes a combination of a compound provided herein and another therapy (e.g., a prophylactic or therapeutic agent) that has been used or is currently being used to prevent, manage, or treat a disorder, which is more effective than the additive effect of the therapies. A synergistic effect of a combination of therapies (e.g., a combination of prophylactic or therapeutic agents) may allow for the use of lower dosages of one or more therapies and / or less frequent administration of the therapies to a mammal with a disorder. The ability to utilize lower dosages of a therapy (e.g., a prophylactic or therapeutic agent) and / or administer the therapy less frequently reduces the toxicity associated with administering the therapy to a mammal without reducing the effectiveness of the therapy in preventing or treating the disorder. Furthermore, a synergistic effect may improve the efficacy of agents in preventing or treating a disorder. Finally, a synergistic effect of a combination of therapies (e.g., a combination of prophylactic or therapeutic agents) may avoid or reduce adverse or unwanted side effects associated with the use of either therapy alone.

[0345] The active compounds provided herein may be administered in combination with or in place of another therapeutic agent, particularly an agent effective in treating wounds and / or Wnt transcription-associated disorders and / or conditions associated with Wnt transcripts and / or Wnt signaling pathway activity. Combination therapy involves administering effective dosages of two or more agents together, while alternation or sequential-step therapy involves administering effective dosages of each agent serially or sequentially. The dosage given will depend on the rate of absorption, inactivation, and excretion of the drug, as well as other factors known to those skilled in the art. It should be noted that dosage values ​​will also vary depending on the severity of the wound or Wnt transcription-associated disorder being alleviated. Furthermore, it should be understood that for any particular mammal, specific dosing regimens and schedules should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions. [Example]

[0346] Example The following examples are provided for the purpose of illustrating certain aspects of the present disclosure and should not be construed as limiting the disclosure in any way.

[0347] As used herein, the symbols and conventions used in these processes, schemes, and examples are consistent with those used in modern scientific literature, e.g., the Journal of the American Chemical Society or the Journal of Biological Chemistry, regardless of whether a particular abbreviation is specifically defined. Specifically, but not by way of limitation, the following abbreviations may be used in the examples and throughout the specification: g (gram); mg (milligram); mL (milliliter); μL (microliter); mM (millimolar); μM (micromolar); Hz (Hertz); MHz (Megahertz); M (Molar); M+1 (MS peak, presence of carbon-13 isotope in molecular ion + 1 peak); mmol (millimolar); m / z (mass per charge); h, hr, or hrs (hours); min (minutes); eq (equivalent); RT, RT, or rt (room temperature); R t or Rt (retention time); R f or Rf (retention factor); v (V) or vol (volume); E (cis); Z (trans); MS (mass spectrometry); ESI (electrospray ionization); TLC (thin layer chromatography); HPLC (high pressure liquid chromatography); LC-MS (liquid chromatography-mass spectrometry); 1H NMR (proton nuclear magnetic resonance); ACN or CH3CN (acetonitrile); Ac2O (acetic anhydride); AcOH (acetic acid); BPin (bis(pinacolato)); BBr3 (boron tribromide); CDCl3 (deuterated chloroform); CH2Cl2 or DCM (dichloromethane); CuBr2 (copper(II) bromide); CN (cyanide or cyano); Cs2CO3 (cesium carbonate); DCM (dichloromethane); DMF (dimethylformamide); DMSO (dimethyl sulfoxide); DMSO-d6 (deuterated dimethyl sulfoxide); EtOAc (ethyl acetate); FA (formic acid); H2 (hydrogen gas); HCl (hydrochloride or hydrochloric acid); I2 (iodine); K2CO3 (potassium carbonate); KOAc (potassium acetate); LDA (lithium diisopropyl ether) propylamide; LAH (lithium aluminum hydride); LHMDS or LiHMDS (lithium bis(trimethylsilyl)amide); MeOH (methanol); MeOD (methanol-D); MeMgBr (methylmagnesium bromide); N2 (nitrogen); NaH (sodium hydride); NH2OH (hydroxylamine); Na2SO4 (sodium sulfate); NaHCO3 (sodium bicarbonate); NaHMDS (sodium bis(trimethylsilyl)amide); NaOMe (sodium methoxide); NH3 (ammonia); NH4Cl (ammonium chloride); NH4HCO3 (ammonium bicarbonate); NMP (n-methyl-2-pyrrolidone); OMe (methoxy); Pd / C (palladium on carbon); PE (petroleum ether); Ph (phenyl); -Si(tert-Bu)(Ph)2) and -Si t BuPh2 (tert-butyl-diphenylsilyl); SiO2 (silicon dioxide); THF (tetrahydrofuran); TFA (trifluoroacetic acid); tBuONO (t-butyl nitrite); dppf (diphenylphosphino); TMS (trimethylsilyl); GO-HA (graphene oxide / hyaluronic acid); MWCO (molecular weight cut off); RPM or rpm (revolutions per minute); N (normality) or N (newtons); and CFU (colony forming units).

[0348] Synthesis Example compound 1 Synthesis of (4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)boronic acid (compound 1)

[0349] [ka]

[0350] Synthesis of 2-(4-bromophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0351] [ka]

[0352] Scheme 1A

[0353] [ka]

[0354] A mixture of 4-bromobenzimidamide hydrochloride 1 (22.0 g, 93.41 mmol, 1.0 eq), methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (1.1 g, 102.76 mmol, 1.1 eq), and KCO (38.73 g, 280.23 mmol, 3.0 eq) in MeOH (660 mL) was stirred at 80 °C for 16 h. The mixture was cooled and concentrated in vacuo. The solid was triturated with water (500 mL), then filtered and dried to give 2-(4-bromophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one 2 (26.4 g, 81.7 mmol). LC-MS: C 13 H 11 Calculated BrN2OS: 323.2; Found: 325.0.

[0355] Synthesis of 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0356] [ka]

[0357] Scheme 1B

[0358] [ka]

[0359] A degassed mixture of 2-(4-bromophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (2) (20 g, 61.88 mmol, 1.0 eq), (BPin) (31.43 g, 123.76 mmol, 2.0 eq), Pd(dppf)Cl (2.26 g, 3.09 mmol, 0.05 eq), and KOAc (30.36 g, 309.40 mmol, 5.0 eq) in dioxane (200 mL) was stirred under argon at 100 °C for 16 h. The mixture was concentrated in vacuo. The crude product was purified by silica gel column chromatography (PE: EtOAc = 5:1 to 1:1) to give 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (3) (21.73 g, 58.72 mmol). LC-MS: C 19 H 23 Calculated for BN2O3S: 370.1; Measured: 371.1.

[0360] Scheme 1C

[0361] [ka]

[0362] A solution of 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (3) (10.0 g, 370.27 mmol, 1.0 eq) in 3 M HCl / MeOH (100 mL) was stirred at 25° C. for 16 h. The mixture was concentrated in vacuo to give the crude product, which was triturated with THF (150 mL) under reflux and filtered to give (4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)boronic acid (compound 1). LC-MS: C 13 H 13 Calculated for BN2O3S: 288.1; Measured: 289.1. 1 H NMR (400 MHz, MeOD): δ 7.95 (s, 4H), 3.67 (s, 2H), 3.10 (m, 2H), 3.02 (m, 2H).

[0363] compound 2 Synthesis of 2-oxo-1-((1r,3r)-3-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)cyclobutyl)-2,3-dihydro-1H-benzo[d]imidazole-5-carbonitrile (Compound 2)

[0364] [ka]

[0365] Synthesis of (Z)-tert-butyl (3-hydroxycyclobutyl)carbamate

[0366] [ka]

[0367] Scheme 2A

[0368] [ka]

[0369] A solution of tert-butyl (3-oxocyclobutyl)carbamate (1) (2.5 g, 13.5 mmol, 1.0 eq) in 80 mL of THF was cooled to -78 °C and treated with a 1 N solution of L-selectride in THF (16.2 mL, 16.2 mmol, 1.2 eq). After stirring for 1 h, the reaction was quenched with 5 mL of water and warmed to rt. The reaction mixture was concentrated and purified by silica gel chromatography (EA:PE = 0-50%) to give (Z)-tert-butyl (3-hydroxycyclobutyl)carbamate (2) (1.8 g). 1 H NMR (400 MHz, CDC13): δ(ppm): 4.68 (brs, 1H), 4.05-3.98 (m, 1H), 3.67-3.65 (m, 1H), 2.78-2.75 (m, 2H), 2.08 (brs, 1H), 1.81-1.78 (m, 2H), 1.44 (s, 9H).

[0370] Synthesis of (Z)-3-((tert-butoxycarbonyl)amino)cyclobutyl methanesulfonate

[0371] [ka]

[0372] Scheme 2B

[0373] [ka]

[0374] Methanesulfonyl chloride (1.3 g, 11.6 mmol, 1.2 eq) was added dropwise to a solution of (Z)-tert-butyl (3-hydroxycyclobutyl)carbamate (2) (1.8 g, 9.6 mmol, 1.0 eq) and TEA (1.5 g, 14.8 mmol, 1.5 eq) in dichloromethane (60 mL) at −70° C. The resulting solution was stirred at −70° C. for 2 h, and the mixture was diluted with 100 mL of water. The resulting solution was extracted with dichloromethane (3 × 60 mL), and the organic layers were combined. The resulting mixture was washed with brine (100 mL), dried over anhydrous sodium sulfate, and concentrated to give (Z)-3-((tert-butoxycarbonyl)amino)cyclobutyl methanesulfonate (3) (2.5 g). 1 H NMR (400 MHz, CDC13): δ(ppm):4.75-4.67 (m, 2H), 3.85-3.82 (m, 1H), 2.98 (s, 3H), 2.93-2.88 (m, 2H), 2.22-2.14 (m, 2H), 1.44 (s, 9H).

[0375] Synthesis of (E)-tert-butyl (3-cyanocyclobutyl)carbamate

[0376] [ka]

[0377] Scheme 2C

[0378] [ka]

[0379] (Z)-3-((tert-butoxycarbonyl)amino)cyclobutyl methanesulfonate 3 (1.2 g, 4.5 mmol, 1.0 eq) in DMF (30 mL) was treated with NaCN (665.7 mg, 13.6 mmol, 3.0 eq), the reaction was heated at 120 °C for 15 h, and the mixture was diluted with 50 mL of water. The resulting solution was extracted with EA (2 × 60 mL), and the organic layers were combined. The resulting mixture was washed with brine (60 mL), dried over anhydrous sodium sulfate, and concentrated to give (E)-tert-butyl (3-cyanocyclobutyl)carbamate 4 (730.0 mg). 1 H NMR (400 MHz, CDCl3): δ(ppm): 4.77 (br s, 1H), 4.41-4.39 (m, 1H), 3.06-3.01 (m, 1H), 2.74-2.41 (m, 2H), 2.27-2.24 (m, 2H), 1.45 (s, 9H).

[0380] Synthesis of (E)-tert-butyl (3-(N-hydroxycarbamimidoyl)cyclobutyl)carbamate

[0381] [ka]

[0382] Scheme 2D

[0383] [ka]

[0384] To a solution of (E)-tert-butyl (3-cyanocyclobutyl)carbamate (4) (730.0 mg, 3.7 mmol, 1.0 eq) in EtOH (25 mL) was added aqueous hydroxylamine (2.2 g / 2.0 mL, 33.3 mmol, 9.0 eq) under N. The mixture was heated to 80 °C and stirred for 15 h. The reaction mixture was concentrated and purified by Prep-HPLC to give (E)-tert-butyl (3-(N-hydroxycarbamimidoyl)cyclobutyl)carbamate (5) (520.0 mg). LCMS (ESI): m / z 230.2 [M+H] + .

[0385] Synthesis of (E)-tert-butyl (3-carbamimidoylcyclobutyl)carbamate

[0386] [ka]

[0387] Scheme 2E

[0388] [ka]

[0389] To a solution of (E)-tert-butyl (3-(N-hydroxycarbamimidoyl)cyclobutyl)carbamate (5) (520.0 mg, 2.3 mmol, 1.0 eq) in MeOH (100 mL) was added Raney Ni (200 mg, 2.3 mmol, 1.0 eq) under N. The suspension was degassed under vacuum and purged with H several times. The mixture was stirred under H at 0 °C for 8 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give (E)-tert-butyl (3-carbamimidoylcyclobutyl)carbamate (6) (480.0 mg). LCMS (ESI): m / z 214.2 [M+H] + .

[0390] Synthesis of (E)-tert-butyl ((1r,3r)-3-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)cyclobutyl)carbamate

[0391] [ka]

[0392] Scheme 2F

[0393] [ka]

[0394] To a solution of (E)-tert-butyl (3-carbamimidoylcyclobutyl)carbamate (6) (480.0 mg, 2.3 mmol, 1.0 eq) and methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (7) (431.3 mg, 2.5 mmol, 1.1 eq) in t-BuOH (30 mL), TEA (1.3 g, 12.8 mmol, 5.6 eq) was added in one portion at 15 °C under N. The mixture was heated to 100 °C and stirred for 15 h. The reaction was concentrated in vacuo. The residue was purified by silica gel column chromatography (DCM / MeOH = 100% to 95%) to give (E)-tert-butyl ((1r,3r)-3-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)cyclobutyl)carbamate (8) (630.1 mg). LCMS (ESI): m / z 338.1 [M+H] + .

[0395] Synthesis of (E)-2-((1r,3r)-3-aminocyclobutyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0396] [ka]

[0397] Scheme 2G

[0398] [ka]

[0399] To a stirred solution of 8 (630 mg, 1.9 mmol, 1.0 eq) in DCM (30.0 mL) was added TFA (3.0 mL, 39.5 mmol, 20.8 eq) at 25 °C. The reaction mixture was stirred at 25 °C under a N atmosphere for 5 h. The reaction mixture was concentrated in vacuo to give (E)-2-((1r,3r)-3-aminocyclobutyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one 9 (656.5 mg). LCMS (ESI): m / z 238.0 [M+H] + . 1 H-NMR(400MHz, DMSO-d6): δ(ppm): 8.41 (brs, 3H), 3.75-3.71 (m, 1H), 3.47 (s, 2H), 3.43-3.80 (m, 1H), 2.90-2.87 (m, 4H), 2.66-2.53 (m, 4H).

[0400] Synthesis of (E)-3-nitro-4-(((1r,3r)-3-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)cyclobutyl)amino)benzonitrile

[0401] [ka]

[0402] Scheme 2H

[0403] [ka]

[0404] To a solution of 9 (443.1 mg, 1.9 mmol, 1.0 eq) and 4-fluoro-3-nitrobenzonitrile 10 (248.3 mg, 1.5 mmol, 0.8 eq) in DMF (30 mL) was added CsCO (1.5 g, 4.6 mmol, 2.4 eq) in one portion under N at 15 °C. The reaction mixture was stirred at 15 °C under N for 15 h, and the mixture was diluted with 50 mL of water. The resulting solution was extracted with EA (2 × 60 mL), and the organic layers were combined. The resulting mixture was washed with brine (60 mL), dried over anhydrous sodium sulfate, and concentrated to give (E)-3-nitro-4-(((1r,3r)-3-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)cyclobutyl)amino)benzonitrile (11) (500.9 mg). LCMS (ESI): m / z 384.0 [M+H] + .

[0405] Synthesis of (E)-3-amino-4-(((1r,3r)-3-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)cyclobutyl)amino)benzonitrile

[0406] [ka]

[0407] Scheme 2J

[0408] [ka]

[0409] To a solution of 11 (200.0 mg, 0.52 mmol, 1.0 eq) and TEA (1.0 mL, 7.2 mmol, 13.8 eq) in MeOH (200 mL) was added Pd / C (200 mg) under N. The suspension was degassed under vacuum and purged with H several times. The mixture was stirred under H at 15 °C for 5 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give (E)-3-amino-4-(((1r,3r)-3-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)cyclobutyl)amino)benzonitrile 12 (180.0 mg). LCMS (ESI): m / z 354.2 [M+H] + .

[0410] Scheme 2K

[0411] [ka]

[0412] (E)-3-amino-4-(((1r,3r)-3-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)cyclobutyl)amino)benzonitrile (12) (180.0 mg, 0.51 mmol, 1.0 eq) and TEA (3 mL, 21.7 mmol, 42.6 eq) in DCM (30 mL) were treated with CDI (918.3 mg, 5.7 mmol, 11.2 eq), the reaction was heated to 50 °C for 15 h, and the mixture was diluted with 50 mL of water. The resulting solution was extracted with EA (2 × 60 mL), and the organic layers were combined. The resulting mixture was washed with brine (60 mL), dried over anhydrous sodium sulfate, and the organic layer was concentrated and purified by Prep-HPLC to give (E)-2-oxo-1-((1r,3r)-3-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)cyclobutyl)-2,3-dihydro-1H-benzo[d]imidazole-5-carbonitrile (Compound 2) (102.0 mg). LCMS (ESI): m / z 380.0 [M+H] + . 1 H-NMR(400MHz, DMSO-d6): δ(ppm): 11.40 (s, 1H), 8.05 (d, J=8.4 Hz, 1H), 7.58 (dd, J=8.0 Hz, J=1.2 Hz, 1H), 7.40 (d, J=1.2 Hz, 1H), 5.02-4.92 (m, 1H), 3.47 (s, 2H), 3.31-3.24 (m, 1H), 3.10-3.02 (m, 2H), 2.93-2.85 (m, 4H), 2.71-2.63 (m, 2H).

[0413] スキーム2L

[0414]

change

[0415] (Compound 2) (150 mg) was purified by HPLC, (Compound 2-P1) and (Compound 2-P2) were obtained. LCMS (ESI): m / z 380.0 [M+H] + . 1 H-NMR (Compound 2-P1, 400 MHz, DMSO-d6): δ (ppm): 7.49 (s, 2H), 7.38 (s, 1H), 5.10-5.04 (m, 1H), 3.58-3.50 (m, 1H), 3.47 (s, 2H), 3.17-3.09 (m, 2H), 2.93-2.85 (m, 4H), 2.71-2.63 (m, 2H). 1H-NMR (Compound 2-P2, 400MHz, DMSO-d6): δ(ppm): 12.49 (s, 1H), 11.39 (s, 1H), 8.09 (d, J=8.4 Hz, 1H), 7.58 (dd, J=8.4 Hz, J=1.6 Hz, 1H), 7.40 (d, J=1.6 Hz, 1H), 5.02-4.92 (m, 1H), 3.47 (s, 2H), 3.31-3.24 (m, 1H), 3.10-3.01 (m, 2H), 2.93-2.90 (m, 4H), 2.71-2.63 (m, 2H).

[0416] compound 3 Synthesis of 2-(1-hydroxy-1,3-dihydrobenzo[c][1,2]oxaborol-5-yl)-7,8-dihydro-3H-thiopyrano[4,3-d]pyrimidin-4(5H)-one (Compound 3)

[0417] [ka]

[0418] Synthesis of methyl 2-bromo-5-cyanobenzoate

[0419] [ka]

[0420] Scheme 3A

[0421] [ka]

[0422] A mixture of CuBr (3.04 g, 13.6 mmol, 1.2 eq) in HCl (50 mL) was added to tBuONO (1.64 g, 15.9 mmol, 1.4 eq) at 0 °C. The mixture was stirred for 5 min. Methyl 2-amino-5-cyanobenzoate 1 (2.0 g, 11.4 mmol, 1 eq) was added in portions. The mixture was stirred at room temperature for 16 h and acidified (pH = 2) by the addition of 1 M HCl. The mixture was extracted with EtOAc (3 × 80 mL), and the combined organic extracts were dried over NaSO, filtered, and evaporated to give methyl 2-bromo-5-cyanobenzoate 2 (2.7 g, crude).

[0423] Synthesis of methyl 2-bromo-5-carbamimidoylbenzoate

[0424] [ka]

[0425] Scheme 3B

[0426] [ka]

[0427] A mixture of methyl 2-bromo-5-cyanobenzoate 2 (2.50 g, 10.4 mmol, 1.0 eq), ammonium chloride (0.38 g, 5.4 mmol, 2.5 eq), and sodium methoxide (0.22 g, 5.4 mmol, 2.5 eq) in MeOH (50 mL) was stirred for 16 h at 40° C. The mixture was concentrated to give methyl 2-bromo-5-carbamimidoylbenzoate 3 (crude). LC-MS (ESI) m / z C9H9BrN2O2+H + Calculated value: 257.1; Measured value: 256.9.

[0428] Synthesis of methyl 2-bromo-5-(4-oxo-4,5,7,8-tetrahydro-3H-thiopyrano[4,3-d]pyrimidin-2-yl)benzoate

[0429] [ka]

[0430] Scheme 3C

[0431] [ka]

[0432] A mixture of methyl 2-bromo-5-carbamimidoylbenzoate (3) (2.5 g, 9.7 mmol, 1.0 eq), methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (1.7 g, 9.7 mmol, 1.0 eq), and KCO (4.0 g, 29.2 mmol, 3.0 eq) in MeOH (50 mL) was stirred at 70 °C for 16 h. The mixture was cooled and filtered. The filtrate was added to water (300 mL). The solid was filtered and dried to give methyl 2-bromo-5-(4-oxo-4,5,7,8-tetrahydro-3H-thiopyrano[4,3-d]pyrimidin-2-yl)benzoate (4) (1.5 g, 3.9 mmol). LC-MS (ESI) m / z C 15 H 13 BrN2O3S+H + Calculated value: 382.2; Measured value: 383.0.

[0433] Synthesis of methyl 5-(4-oxo-4,5,7,8-tetrahydro-3H-thiopyrano[4,3-d]pyrimidin-2-yl)-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate

[0434] [ka]

[0435] Scheme 3D

[0436] [ka]

[0437] A mixture of methyl 2-bromo-5-(4-oxo-4,5,7,8-tetrahydro-3H-thiopyrano[4,3-d]pyrimidin-2-yl)benzoate (4) (0.60 g, 1.57 mmol, 1.0 eq), (BPin) (0.80 g, 3.15 mmol, 2.0 eq), Pd(dppf)Cl (115 mg, 0.16 mmol, 0.1 eq), and KOAc (462 mg, 4.72 mmol, 3.0 eq) in dioxane (15 mL) was stirred at 110 °C for 16 h. The mixture was diluted with EtOAc (30 mL), washed with water (15 mL), dried over NaSO, and concentrated. The crude product was purified by silica gel column chromatography (PE: EtOAc = 5:1 to 1:1) to give methyl 5-(4-oxo-4,5,7,8-tetrahydro-3H-thiopyrano[4,3-d]pyrimidin-2-yl)-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (5) (340 mg, 0.79 mmol). LC-MS (ESI) m / z C 21 H 25 BN2O5S+H + Calculated value: 429.3; Measured value: 429.1.

[0438] Scheme 3E

[0439] [ka]

[0440] To a solution of methyl 5-(4-oxo-4,5,7,8-tetrahydro-3H-thiopyrano[4,3-d]pyrimidin-2-yl)-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (5) (0.34 g, 0.79 mmol, 1.0 eq) in THF (15 mL) was added LAH (60 mg, 1.60 mmol, 2.0 eq). The mixture was stirred at 0 °C for 1 h. The mixture was added to HO, filtered, and concentrated in vacuo to give the crude product, which was purified by flash chromatography (HO:CHCN = 90:10 to 50:50) to give 2-(1-hydroxy-1,3-dihydrobenzo[c][1,2]oxaborol-5-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 3). LC-MS (ESI) m / z C 14 H 13 BN2O3S +H + Calculated value: 301.1; Measured value: 301.0. 1 H NMR (400 MHz, MeOD): δ 8.09 (s, 1H), 8.03 (d, J = 7.6 Hz, 1H), 7.84 (d, J = 7.6 Hz, 1H), 3.54 (s, 2H), 5.07 (s, 2H), 2.95-2.85 (m, 4H).

[0441] Compound 4 and Compound 43 Synthesis of 3-(4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)-1,2,4-oxadiazol-5(4H)-one (Compound 4)

[0442] [ka]

[0443] Synthesis of 4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)benzonitrile (Compound 43)

[0444] [ka]

[0445] Synthesis of 2-(4-bromophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0446] [ka]

[0447] Scheme 4A

[0448] [ka]

[0449] To a solution of methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (1) (5.00 g, 28.7 mmol, 1.00 eq) and 4-bromobenzamidine hydrochloride (6.76 g, 28.7 mmol, 1.00 eq) in EtOH (50 mL) was added KCO (7.93 g, 57.4 mmol, 2 eq) at 20 °C. The mixture was stirred at 80 °C for 16 h. LC-MS showed that 1 was completely consumed. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue, which was suspended in water (100 mL) and then stirred at 20 °C for 4 h. The mixture was filtered, and the filter cake was dried under reduced pressure to give 2-(4-bromophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (2) (6.60 g, crude), which was used directly in the next step without further purification. 1 H NMR (400 MHz, DMSO-d6): δ 8.08 - 8.02 (m, 2H), 7.72 - 7.67 (m, 2H), 3.51 (s, 2H), 2.87 (qd, J = 4.4, 8.4 Hz, 4H). LC-MS: 324.3 + bromo isomer (M+1).

[0450] Synthesis of 4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)benzonitrile (Compound 43)

[0451] [ka]

[0452] Scheme 4B

[0453] [ka]

[0454] To a solution of 2-(4-bromophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (2) (4.50 g, 13.9 mmol, 1 eq) in NMP (45 mL) was added Zn(CN) (981 mg, 8.35 mmol, 530 μL, 0.6 eq) and Pd(PPh) (1.61 g, 1.39 mmol, 0.1 eq) in portions at 20 °C under N. The mixture was stirred at 100 °C for 2 h. LC-MS showed that 2 was completely consumed. The resulting mixture was cooled to 20 °C and diluted with saturated NaCO (120 mL) at 20 °C. The mixture was then extracted with EtOAc (40 mL × 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue, which was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1) to give 4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)benzonitrile (Compound 43) (1.80 g, 6.68 mmol). 1H NMR (400 MHz, DMSO-d6): δ 13.07 - 12.77 (m, 1H), 8.25 (br d, J = 7.5 Hz, 2H), 8.00 (d, J = 8.4 Hz, 2H), 3.55 (s, 2H), 2.91 (s, 4H). LC-MS: 270.1 (M+1).

[0455] Synthesis of (Z)-N'-hydroxy-4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)benzimidamide

[0456] [ka]

[0457] Scheme 4C

[0458] [ka]

[0459] To a solution of 4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)benzonitrile (compound 43) (500 mg, 1.86 mmol, 1 eq) in pyridine (5 mL) was added NHOH.HCl (258 mg, 3.71 mmol, 2 eq) and KCO (641 mg, 4.64 mmol, 2.50 eq) at 20 °C. The mixture was stirred at 100 °C for 13 h. LC-MS showed that (compound 43) was consumed. After cooling to 20 °C, the reaction mixture was filtered, and the filter cake was dried and concentrated under reduced pressure to give a residue. This residue was suspended in water (5.00 mL) and stirred at 20 °C for 4 h. The mixture was filtered, and the filter cake was dried and concentrated under reduced pressure to give (Z)-N'-hydroxy-4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)benzimidamide (4) (333 mg, crude), which was used in the next step without further purification. LC-MS: 302.9 (M+1).

[0460] Scheme 4D

[0461] [ka]

[0462] To a solution of (Z)-N'-hydroxy-4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)benzimidamide (4) (333 mg, 1.10 mmol, 1 eq) in pyridine (3.30 mL) was added CDI (268 mg, 1.65 mmol, 1.50 eq) at 20 °C. The mixture was stirred at 110 °C for 3 h. LC-MS showed that 4 was completely consumed. After cooling to 20 °C, the mixture was filtered, and the filter cake was dried and concentrated under reduced pressure to give a residue, which was stirred in DCM (2 mL) at RT for 6 h. The mixture was filtered, and the filter cake was dried and concentrated under reduced pressure to give 3-(4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)-1,2,4-oxadiazol-5(4H)-one (compound 4). 1 H NMR (400 MHz, DMSO-d6): δ 13.26 - 12.46 (m, 1H), 8.23 ​​(d, J = 8.5 Hz, 2H), 7.94 (d, J = 8.5 Hz, 2H), 7.18 (s, 1H), 3.55 (s, 2H), 2.91 (s, 4H). LC-MS: 329.1 (M+1).

[0463] compound 5 Synthesis of (E)-(5-(2-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)vinyl)-2-(trifluoromethyl)phenyl)boronic acid (compound 5)

[0464] [ka]

[0465] Synthesis of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(trifluoromethyl)benzaldehyde

[0466] [ka]

[0467] Scheme 5A

[0468] [ka]

[0469] To a solution of 3-bromo-4-(trifluoromethyl)benzaldehyde (1) (0.50 g, 1.98 mmol, 1.00 eq) and Pin2B2 (753 mg, 2.96 mmol, 1.50 eq) in DMSO (10.0 mL) was added KOAc (1.36 g, 13.8 mmol, 7.00 eq) and Pd(dppf)Cl.CHCl (161 mg, 198 μmol, 0.10 eq) under N at 25 °C. The suspension was degassed in vacuo and purged with N several times. The mixture was then warmed to 80 °C and stirred at 80 °C for 2 h. Thin-layer chromatography (TLC; petroleum ether / ethyl acetate = 20 / 1) showed that 3-bromo-4-(trifluoromethyl)benzaldehyde (1) was completely consumed. The mixture was cooled to 25°C and then diluted with EtOAc (5.00 mL). The mixture was filtered, and the filtrate was washed with water (5.00 mL x 2) and brine (5.00 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO2, petroleum ether / ethyl acetate = 10 / 1). 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(trifluoromethyl)benzaldehyde (2) (0.20 g, crude) was obtained, which was used in the next step without further purification.

[0470] Synthesis of 2-methyl-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0471] [ka]

[0472] Scheme 5B

[0473] [ka]

[0474] To a solution of acetamidine hydrochloride 4 (814 mg, 8.61 mmol, 1.50 eq) in MeOH (10.0 mL) was added KCO (1.98 g, 14.4 mmol, 2.50 eq) and methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate 3 (1.00 g, 5.74 mmol, 1.00 eq). The mixture was stirred at 20 °C for 12 h. TLC (petroleum ether / ethyl acetate = 8 / 1) showed that methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate 3 was completely consumed. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. 2-Methyl-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (1.00 g, crude) was obtained, which was used in the next step without further purification. 1 Confirmed by 1 H NMR.

[0475] Synthesis of (E)-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(trifluoromethyl)styryl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0476] [ka]

[0477] Scheme 5C

[0478] [ka]

[0479] To a solution of 2-methyl-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one 5 (0.20 g, 666 μmol, 1.21 eq) in AcO (1.00 mL) was added 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(trifluoromethyl)benzaldehyde 2 (0.10 g, 549 μmol, 1.10 g, 549 μmol, 1.00 eq), ZnCl (150 mg, 1.10 mmol, 51.4 μL, 2.00 eq), and AcOH (65.9 mg, 1.10 mmol, 62.8 μL, 2.00 eq) under N at 25 °C. The mixture was stirred at 120 °C for 4 h. LC-MS (ET48116-9-P1A) showed that 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(trifluoromethyl)-benzaldehyde (2) was completely consumed, and one main peak with the desired m / z was detected. The mixture was cooled to 25 °C and then concentrated under reduced pressure to remove most of the AcO. The resulting residue was dissolved in EtOAc (5.00 mL) and then washed with saturated NaHCO (2.00 mL × 2). The organic layer was separated and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO, petroleum ether / ethyl acetate = 1 / 1). (E)-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(trifluoromethyl)styryl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (6) (0.25 g, crude) was obtained.

[0480] Scheme 5D

[0481] [ka]

[0482] A mixture of (E)-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(trifluoromethyl)styryl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (6) (0.25 g, 538.44 μmol, 1 eq) in HCl / HO (4 M, 2.50 mL) was stirred at 80 °C for 2 h. LC-MS showed that starting material (6) was completely consumed. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by prep-HPLC (column: Phenomenex Luna C18 80 × 40 mm × 3 μm; mobile phase: [water (HCl)-ACN]; B%: 18% to 48%, 7 min). (E)-(5-(2-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)vinyl)-2-(trifluoromethyl)phenyl)boronic acid (compound 5) was obtained. LC-MS: 383.0 (M+1). 1 H NMR (400MHz, DMSO-d6): δ= 7.89 (br d, J=16.4 Hz, 1H), 7.80 - 7.66 (m, 3H), 7.04 (br d, J=16.3 Hz, 1H), 3.51 (br s, 2H), 2.93 - 2.81 (m, 4H).

[0483] compound 6 Synthesis of (E)-2-(4-(trifluoromethyl)styryl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (Compound 6)

[0484] [ka]

[0485] Synthesis of 2-methyl-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0486] [ka]

[0487] Scheme 6A

[0488] [ka]

[0489] To a solution of acetamidine hydrochloride (814 mg, 8.61 mmol, 1.50 eq) in MeOH (10.0 mL) were added K2CO3 (1.98 g, 14.4 mmol, 2.50 eq) and methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate 1 (1.00 g, 5.74 mmol, 1.00 eq). The mixture was stirred at 20 °C for 12 h. TLC (petroleum ether / ethyl acetate = 8 / 1) showed that starting material 1 was completely consumed. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. 2-Methyl-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one 2 (1.00 g, crude) was obtained, which was used in the next step without further purification.

[0490] Scheme 6B

[0491] [ka]

[0492] To a solution of 2-methyl-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (2) (0.25 g, 1.37 mmol, 1.00 eq) in AcO (2.50 mL) was added 4-(trifluoromethyl)benzaldehyde (244 mg, 1.40 mmol, 187 μL, 1.02 eq) at 20 °C. The mixture was stirred at 110 °C for 12 h. The mixture was cooled to 20 °C and concentrated under reduced pressure to give a residue. The mixture was diluted with H2O (2.00 mL) and extracted with EtOAc (2.00 mL × 3). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give a residue. A portion of the residue was purified by prep-HPLC (Waters Xbridge BEH C18 100 × 30 mm × 10 μm column; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 40% to 65%, 10 min) to give (E)-2-(4-(trifluoromethyl)styryl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 6). LCMS: 339.1 (M+1). 1 H NMR (400MHz, DMSO-d6): δ= 12.62 - 12.51 (m, 1H), 7.91 - 7.73 (m, 5H), 7.04 - 6.92 (m, 1H), 3.52 - 3.45 (m, 2H), 2.89 - 2.79 (m, 4H).

[0493] compound 7 Synthesis of 2-(4-(6-bromopyridin-3-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 7)

[0494] [ka]

[0495] Synthesis of 4-(6-bromopyridin-3-yl)benzonitrile

[0496] [ka]

[0497] Scheme 7A

[0498] [ka]

[0499] A mixture of (4-cyanophenyl)boronic acid 1 (50.0 g, 340.0 mmol, 1.0 eq), 2-bromo-5-iodopyridine (96.6 g, 340.2 mmol, 1.0 eq), and KCO (141.0 g, 1.02 mol, 3.0 eq) in dioxane (3.2 L) and HO (800 mL) was stirred at 60 °C for 16 h. The mixture was filtered and concentrated. The crude residue was purified on a silica gel column (PE: EtOAc = 10:1 to 1:1) to give 4-(6-bromopyridin-3-yl)benzonitrile 2 (63.0 g). LC-MS (ESI) m / z C 12 H7BrN2+H + Calculated value: 261.0; Measured value: 261.1 ( 81 isotope of Br). This reaction is shown in Scheme 7A.

[0500] Synthesis of 4-(6-bromopyridin-3-yl)-N-hydroxybenzimidamide

[0501] [ka]

[0502] Scheme 7B

[0503] [ka]

[0504] A mixture of 4-(6-bromopyridin-3-yl)benzonitrile 2 (60 g, 231.6 mmol, 1.0 eq), NHOH HCl (40.2 g, 579 mmol, 2.5 eq), and NaOH (23.16 g, 579 mmol, 2.5 eq) in EtOH (500 mL) was stirred at 80 °C for 3 h. The mixture was diluted with EtOH (100 mL) and filtered. The filtrate was concentrated to give 4-(6-bromopyridin-3-yl)-N-hydroxybenzimidamide 3 (40 g, crude). LC-MS (ESI) m / z C 12 H 10 BrNO+H + Calculated value: 294.0; Measured value: 294.1 ( 81 isotope of Br). This reaction is shown in Scheme 7B.

[0505] Synthesis of 4-(6-bromopyridin-3-yl)benzimidamide

[0506] [ka]

[0507] Scheme 7C

[0508] [ka]

[0509] A mixture of 4-(6-bromopyridin-3-yl)-N-hydroxybenzimidamide (3) (40.0 g, 136.8 mmol, 1.0 eq), NH4Cl (146.48 g, 3.460 mol, 20 eq), and Fe (230.04 g, 2.74 mol, 20 eq) in EtOH (600 mL) was stirred at 80 °C for 48 h. The mixture was cooled and filtered. The solvent was removed in vacuo to give 4-(6-bromopyridin-3-yl)benzimidamide (4) (30.0 g, crude). LC-MS (ESI) m / z C 12 H 10 BrN3+H+ Calculated value: 276.0; Measured value: 276.1 ( 79 isotope of Br). This reaction is shown in Scheme 7C.

[0510] Scheme 7D

[0511] [ka]

[0512] A mixture of 4-(6-bromopyridin-3-yl)benzimidamide (4) (30.0 g, 108.6 mmol, 1.0 eq), methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (37.8 g, 217.2 mmol, 2 eq), and KCO (45.0 g, 325.8 mmol, 3.0 eq) in MeOH (500 mL) was stirred at 70 °C for 4 h. The mixture was concentrated under reduced pressure. The residue was washed with HO (200 mL) and filtered. The solid was triturated in MeOH (150 mL), EtOAc (150 mL), and acetone (100 mL), respectively, to remove most of the impurities. The mixture was filtered. The crude solid was triturated in aqueous NaOH (1% wt, 20 mL) and filtered. The solid was washed with water (50 mL) and acetone (100 mL) and dried under vacuum to give 2-(4-(6-bromopyridin-3-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 7). LC-MS (ESI) m / z C 18 H 14 BrN3OS+H + Calculated value: 402.0; Measured value: 401.9 ( 81 isotope of Br). 1H NMR (400 MHz, DMSO): δ 12.82 (brs, 1H), 8.81 (d, J = 2.4 Hz, 1H), 8.22 (d, J = 8.3 Hz, 2H), 8.14 (dd, J = 8.4, 2.4 Hz, 1H), 7.91 (d, J = 8.3 Hz, 2H), 7.77 (d, J = 8.4 Hz, 1H), 3.54 (s, 2H), 2.91 (brs, 4H). This reaction is shown in Scheme 7D.

[0513] compound 8 Synthesis of 2-(4-(2-(2-hydroxyethoxy)propan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 8)

[0514] [ka]

[0515] Synthesis of methyl 4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)benzoate

[0516] [ka]

[0517] Scheme 8A

[0518] [ka]

[0519] To a solution of methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (1) (2 g, 11.48 mmol, 1 eq) in MeOH (20 mL) was added methyl 4-carbamimidoylbenzoate hydrochloride (1a) (3.20 g, 14.91 mmol, 1.30 eq) and KCO (4.00 g, 28.94 mmol, 2.52 eq) at 25 °C. The mixture was stirred at 25 °C for 12 h. The mixture was filtered, and the filter cake was concentrated under reduced pressure to give a residue. The residue was suspended in HO (40 mL) and stirred at 25 °C for 12 h. The mixture was filtered, and the filter cake was dried in vacuo to give methyl 4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)benzoate (2) (1.5 g, crude). The crude product was used directly in the next step without further purification. This reaction is shown in Scheme 8A.

[0520] Synthesis of 2-(4-(2-hydroxypropan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0521] [ka]

[0522] Scheme 8B

[0523] [ka]

[0524] To a solution of methyl 4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)benzoate (2) (0.3 g, 992.24 μmol, 1 eq) in THF (3 mL) was added MeMgBr (3 M, 992.24 μL, 3 eq) dropwise at 0 °C under N. The mixture was stirred at 25 °C for 3 h. The reaction mixture was quenched by adding NH.sub.4Cl (5 mL), and the aqueous phase was extracted with 10 mL of DCM (5 mL × 2). The combined organic layers were concentrated under reduced pressure to give a residue. 2-(4-(2-hydroxypropan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (3) (2 g, crude) was obtained, which was used directly in the next step without further purification. 1 H NMR: ET47629-1-P1A (400 MHz, CDCl3): δ 12.54 - 12.27 (m, 1H), 8.07 (d, J = 8.5 Hz, 2H), 7.56 (d, J = 8.5 Hz, 2H), 3.62 (br s, 2H), 3.03 - 2.94 (m, 2H), 2.91 - 2.82 (m, 2H), 1.55 (s, 6H). This reaction is shown in Scheme 8B.

[0525] Scheme 8C

[0526] [ka]

[0527] To a solution of 2-(4-(2-hydroxypropan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (3) (0.12 g, 396.84 μmol, 1 eq) in ethylene glycol (5 mL) was added TosOH (82.00 mg, 476.21 μmol, 1.2 eq) at 20 °C. The mixture was stirred at 20 °C for 1 h, then warmed to 80 °C and stirred at 80 °C for 12 h. The mixture was directly purified by prep-HPLC (neutral conditions, column: Waters Xbridge BEH C18 100 × 30 mm × 10 μm; mobile phase: [water (NH4HCO3)-ACN]; B%: 10% to 50%, 10 min). 2-(4-(2-(2-hydroxyethoxy)propan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 8) was obtained. LC-MS: 347.2 (M +1). 1 H NMR: ET47430-4-P1A1 (400 MHz, CDCl3): δ 8.14 (d, J = 8.2 Hz, 2H), 7.60 (d, J = 8.3 Hz, 2H), 3.80 - 3.69 (m, 4H), 3.34 (t, J = 4.6 Hz, 2H), 3.14 - 3.06 (m, 2H), 3.01 - 2.93 (m, 2H), 1.62 (s, 6H). This reaction is shown in Scheme 8C.

[0528] compound 9 Synthesis of 2-(4'-bromo-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 9)

[0529] [ka]

[0530] Synthesis of 4'-bromo-[1,1'-biphenyl]-4-carboximidamide

[0531] [ka]

[0532] Scheme 9A

[0533] [ka]

[0534] To a mixture of 4'-bromo-[1,1'-biphenyl]-4-carbonitrile (1) (4 g, 15.5 mmol, 1 eq) in THF (40 mL) was added dropwise NaHMDS (1 M, 18.60 mL, 1.2 eq) under N2 at 25 °C. The mixture was stirred at 25 °C for 1 h to give a liquid. The reaction mixture was quenched by adding water (30 mL) at 25 °C. The pH of the mixture was adjusted to 2-3 by adding 1 M HCl. The aqueous layer was separated and evaporated to approximately 15% of its original volume, allowing the desired HCl salt to crystallize. The mixture was filtered, and the filter cake was dried in vacuo to give 4'-bromo-[1,1'-biphenyl]-4-carboximidamide (2) (4.53 g, 14.5 mmol, HCl). The crude product was used in the next step without further purification. 1 H NMR: (400 MHz, DMSO-d6): δ 9.44 (s, 2H), 9.19 (s, 2H), 7.94 (s, 4H), 7.81 - 7.65 (m, 4H).

[0535] Scheme 9B

[0536] [ka]

[0537] To a solution of 4'-bromo-[1,1'-biphenyl]-4-carboximidamide (2) (0.066 g, 378.8 μmol, 1.1 eq) in MeOH (1.8 mL) was added methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (3) (107.3 mg, 344.4 μmol, 1 eq, HCl) and KCO (95.2 mg, 688.8 μmol, 2 eq) at 25 °C. The mixture was stirred at 25 °C for 16 h. The reaction mixture was filtered, and the filter cake was dried in vacuo to give a residue. The crude product was purified by precipitation from DMSO (3 mL) to give 2-(4'-bromo-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 9). LC-MS: 399.19 (M+1). 1 H NMR: ET48394-6-P1A (400 MHz, DMSO-d6): δ 12.86 - 12.62 (m, 1H), 8.21 (d, J = 8.3 Hz, 2H), 7.86 - 7.81 (m, 2H), 7.75 - 7.68 (m, 4H), 3.55 (s, 2H), 2.91 (s, 4H).

[0538] compound 10 Synthesis of (4'-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1'-biphenyl]-4-yl)boronic acid (compound 10)

[0539] [ka]

[0540] Synthesis of 2-(4'-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0541] [ka]

[0542] Scheme 10A

[0543] [ka]

[0544] To a mixture of 2-(4'-bromo-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 9) (0.31 g, 776.4 μmol, 1 eq) and B2Pin2 (394.3 mg, 1.55 mmol, 2 eq) in dioxane (6 mL), KOAc (152.4 mg, 1.55 mmol, 2 eq) and Pd(dppf)Cl2.CHCl2 (63.4 mg, 77.6 μmol, 0.1 eq) were added in portions under N2 at 25 °C. The mixture was degassed under vacuum and purged with N2 three times. The reaction mixture was then heated to 100 °C and stirred for 16 h. A suspension was obtained. The crude product was triturated with HO (10 mL) at 25 °C for 2 h. The mixture was filtered, and the filter cake was washed with DCM (5 mL) and MeOH (5 mL). The filter cake was dried under reduced pressure to give 2-(4'-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (0.18 g), which was used in the next step without further purification.

[0545] Scheme 10B

[0546] [ka]

[0547] To a round-bottom flask was added 2-(4'-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (0.18 g, 403.2 μmol, 1 eq) and HCl (4 M, 4 mL, 39.7 eq) at 25 °C. The reaction mixture was stirred at 80 °C for 4 h. The mixture was filtered, and the filter cake was washed with DCM (5 mL) and MeOH (5 mL). The filter cake was dried in vacuo to give (4'-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1'-biphenyl]-4-yl)boronic acid (compound 10). LC-MS: 365.1 (M+1). 1 H NMR: ET48394-20 (400 MHz, DMSO-d6): δ 8.20 (d, J = 8.5 Hz, 2H), 7.89 (dd, J = 8.3, 19.4 Hz, 4H), 7.74 (d, J = 8.2 Hz, 2H), 3.55 (s, 2H), 2.92 (s, 4H).

[0548] compound 11 Synthesis of 2-(4-(2-(2-hydroxyethoxy)pyrimidin-5-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (Compound 11)

[0549] [ka]

[0550] Synthesis of 4-(2-chloropyrimidin-5-yl)benzonitrile

[0551] [ka]

[0552] Scheme 11A

[0553] [ka]

[0554] To a mixture of (2-chloropyrimidin-5-yl)boronic acid (2) (9.57 g, 60.43 mmol, 1.10 eq) and 4-bromobenzonitrile (1) (10.0 g, 54.9 mmol, 1.00 eq) in dioxane (200 mL) and HO (20.0 mL), KCO (15.2 g, 110 mmol, 2.00 eq) and Pd(dppf)Cl.CHCl (4.49 g, 5.49 mmol, 0.10 eq) were added in one portion under N at 25 °C. The mixture was degassed with N and then stirred at 100 °C for 16 h. A liquid was obtained. The reaction mixture was diluted with water (200 mL) and extracted with 600 mL of EtOAc (3 x 200 mL). The combined organic layers were concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (ISCO®; 200 g SepaFlash® silica flash column, eluent: 0–20% ethyl acetate / petroleum ether gradient at 100 mL / min, R f =0.25) to give 4-(2-chloropyrimidin-5-yl)benzonitrile (3) (1.1 g), which was used crude in the next step.

[0555] Synthesis of 4-(2-(2-hydroxyethoxy)pyrimidin-5-yl)benzonitrile

[0556] [ka]

[0557] Scheme 11B

[0558] [ka]

[0559] To a mixture of 4-(2-chloropyrimidin-5-yl)benzonitrile (3) (0.70 g, 3.25 mmol, 1.00 eq) in THF (4.00 mL) and ethylene glycol (4.00 mL) was added KCO (897 mg, 6.49 mmol, 2.00 eq) in one portion at 25 °C under N. The mixture was stirred at 70 °C for 16 h. A suspension was obtained. The reaction mixture was diluted with water (20 mL) and extracted with 30 mL of EtOAc (10 mL × 3). The combined organic layers were concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® silica flash column, eluting with a 0–50% ethyl acetate / petroleum ether gradient at 40 mL / min, R f =0.27) to give 4-(2-(2-hydroxyethoxy)pyrimidin-5-yl)benzonitrile (4) (540 mg, 2.24 mmol), which was used crude in the next step.

[0560] Synthesis of (Z)-N'-hydroxy-4-(2-(2-hydroxyethoxy)pyrimidin-5-yl)benzimidamide

[0561] [ka]

[0562] Scheme 11C

[0563] [ka]

[0564] To a mixture of 4-(2-(2-hydroxyethoxy)pyrimidin-5-yl)benzonitrile 4 (0.54 g, 2.24 mmol, 1.00 eq) in MeOH (5.00 mL) was added NHOH.HCl (171 mg, 2.46 mmol, 1.10 eq) and NaHCO (207 mg, 2.46 mmol, 95.8 μL, 1.10 eq) in one portion at 25 °C under N. The mixture was stirred at 65 °C for 16 h. A liquid was obtained. The reaction mixture was filtered and concentrated under reduced pressure to give (Z)-N'-hydroxy-4-(2-(2-hydroxyethoxy)pyrimidin-5-yl)benzimidamide 5 (500 mg, crude), which was used in the next step without further purification.

[0565] Synthesis of 4-(2-(2-hydroxyethoxy)pyrimidin-5-yl)benzimidamide

[0566] [ka]

[0567] Scheme 11D

[0568] [ka]

[0569] To a mixture of (Z)-N'-hydroxy-4-(2-(2-hydroxyethoxy)pyrimidin-5-yl)benzimidamide (5) (0.50 g, 1.82 mmol, 1.00 eq) in AcOH (2.62 g, 43.7 mmol, 2.50 mL) was added AcO (744 mg, 7.29 mmol, 683 μL, 4.00 eq) in one portion at 25 °C under N. The mixture was stirred at 25 °C for 30 min. Then MeOH (20 mL) and Pd / C (0.2 g, 218.76 μmol) were added. The resulting mixture was degassed three times with H and stirred at 25 °C under H (15 psi) for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give 4-(2-(2-hydroxyethoxy)pyrimidin-5-yl)benzimidamide (6) (500 mg, crude), which was used in the next step without further purification.

[0570] Scheme 11E

[0571] [ka]

[0572] To a mixture of 4-(2-(2-hydroxyethoxy)pyrimidin-5-yl)benzimidamide (6) (500 mg, 1.94 mmol, 1.00 eq) and methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (7) (438 mg, 2.52 mmol, 1.30 eq) in MeOH (4.00 mL), KCO (535 mg, 3.87 mmol, 2.00 eq) was added in one portion at 25 °C under N. The mixture was stirred at 25 °C for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue, which was purified by prep-HPLC (Column: Waters Xbridge Prep OBD C18 150 × 40 mm × 10 μm; Mobile phase: [water (NHHCO)-ACN]; B%: 10% to 40%, 8 min). 2-(4-(2-(2-hydroxyethoxy)pyrimidin-5-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 11) was obtained. 1H NMR (400 MHz, DMSO-d6) δ= 13.15 - 12.49 (m, 1H), 9.03 (s, 2H), 8.21 (br d, J = 8.2 Hz, 2H), 7.91 (d, J = 8.6 Hz, 2H), 4.93 (t, J = 5.5 Hz, 1H), 4.42 - 4.36 (m, 2H), 3.76 (q, J = 5.4 Hz, 2H), 3.54 (s, 2H), 2.95 - 2.86 (m, 4H). LCMS: 383 (M+1).

[0573] compound 12 Synthesis of 2-(4'-(2-(2-hydroxyethoxy)propan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (Compound 12)

[0574] [ka]

[0575] Synthesis of 2-(4-bromophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0576] [ka]

[0577] Scheme 12A

[0578] [ka]

[0579] To a mixture of methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate 1 (8.00 g, 45.9 mmol, 1.00 eq) and 4-bromobenzimidamide 3 (11.9 g, 50.5 mmol, 1.10 eq, HCl) in EtOH (100 mL) was added KCO (12.7 g, 91.8 mmol, 2.00 eq) in one portion at 25 °C under N. The mixture was stirred at 85 °C for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give 2-(4-bromophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one 2 (15.6 g, crude), which was used in the next step without further purification. 1 H NMR: (400 MHz, DMSO-d6): δ 8.19 - 8.13 (m, 2H), 7.53 - 7.46 (m, 2H), 3.42 (s, 2H), 2.81 - 2.75 (m, 2H), 2.74 - 2.67 (m, 2H).

[0580] Synthesis of 2-(4'-(2-hydroxypropan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0581] [ka]

[0582] Scheme 12B

[0583] [ka]

[0584] To a mixture of 2-(4-bromophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (2) (4.00 g, 12.4 mmol, 1.00 eq) and (4-(2-hydroxypropan-2-yl)phenyl)boronic acid (4) (2.67 g, 14.9 mmol, 1.20 eq) in dioxane (40.0 mL) and HO (5.00 mL) was added KCO (3.42 g, 24.8 mmol, 2.00 eq) and Pd(dppf)Cl.CHCl (1.01 g, 1.24 mmol, 0.10 eq) in one portion under N at 25 °C. The mixture was degassed under vacuum and purged with N three times, then heated to 100 °C and stirred for 16 h. The reaction mixture was diluted with EtOAc (20.0 mL) and then filtered. The filter cake was washed with water (10 mL) and MeOH (10 mL) to give 2-(4'-(2-hydroxypropan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (2.10 g, 5.55 mmol, crude), which was used in the next step without further purification. 1 H NMR: (400 MHz, DMSO-d6): δ= 13.13 - 12.31 (m, 1H), 8.51 - 7.75 (m, 4H), 7.75 - 7.15 (m, 4H), 5.07 (br d, J = 3.2 Hz, 1H), 3.54 (br s, 2H), 2.90 (br s, 4H), 1.78 - 0.88 (m, 6H).

[0585] Scheme 12C

[0586] [ka]

[0587] To a mixture of 2-(4'-(2-hydroxypropan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (1.50 g, 3.96 mmol, 1.00 eq) in DMF (10.0 mL) was added TosOH (819 mg, 4.76 mmol, 1.20 eq) and ethylene glycol (5.55 g, 89.4 mmol, 5.00 mL, 22.6 eq) in one portion at 25 °C under N. The mixture was stirred at 85 °C for 16 h. The mixture was cooled to RT and filtered. The filter cake was dissolved in DMSO and directly purified by prep-HPLC (column: Phenomenex Luna 80 × 30 mm × 3 μm; mobile phase: [water (HCl)-MEOH]; B%: 40%–65%, 8 min) to give 2-(4′-(2-(2-hydroxyethoxy)propan-2-yl)-[1,1′-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 12). LCMS: 423.2 (M+1). 1 H NMR: (400 MHz, chloroform-d): δ 11.47 - 10.94 (m, 1H), 8.14 (d, J = 8.4 Hz, 2H), 7.76 (d, J = 8.5 Hz, 2H), 7.67 - 7.61 (m, 2H), 7.54 (d, J = 8.4 Hz, 2H), 3.80 - 3.69 (m, 4H), 3.41 - 3.31 (m, 2H), 3.10 - 3.03 (m, 2H), 2.99 - 2.91 (m, 2H), 1.66 - 1.62 (m, 6H).

[0588] compound 13 Synthesis of 2-(4-(trifluoromethyl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one-8,8-d2 (Compound 13)

[0589] [ka]

[0590] Synthesis of methyl-d3 4-oxotetrahydro-2H-thiopyran-3-carboxylate-3,5,5-d3

[0591] [ka]

[0592] Scheme 13A

[0593] [ka]

[0594] A mixture of methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (1) (0.9 g, 5.16 mmol, 1.0 eq) and NaH (2.1 mg, 0.051 mmol, 0.01 eq) in MeOD (13.5 mL) was stirred at 65 °C for 2 h. Methyl-d3 4-oxotetrahydro-2H-thiopyran-3-carboxylate-3,5,5-d3 (2) was obtained as a crude product in MeOD solution. 1 H NMR (400 MHz, MeOD): δ 3.23-3.07 (m, 1H), 2.95 (s, 1H), 2.76 (s, 1H), 2.73 (s, 2H).

[0595] Scheme 13B

[0596] [ka]

[0597] A mixture of 2 (0.90 g, 5.07 mmol, 1.11 eq) and 4-(trifluoromethyl)benzimidamide (0.861 g, 4.56 mmol, 1.0 eq) in MeOD (12 mL) was stirred at 65 °C for 1 h. The mixture was concentrated to half its volume and then added to HO (50 mL). The precipitate was filtered, washed with HO (30 mL) and MeOH / HO (20 mL, 1:1, v / v), and dried in vacuo to give 2-(4-(trifluoromethyl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one-8,8-d (compound 13). LC-MS (ESI) m / z C 14 H9D2F3N2OS+H + Calculated value: 315.1; Measured value: 315.1. 1 H NMR (400 MHz, DMSO-d6): δ 8.29 (d, J = 8 Hz, 2H), 7.88 (d, J = 8 Hz, 2H), 3.54 (s, 2H), 2.89 (s, 2H). 19 F NMR (375 MHz, DMSO-d6): δ 61.33 (s, 3F).

[0598] compound 14 Synthesis of (2,6-difluoro-4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)boronic acid (compound 14)

[0599] [ka]

[0600] Synthesis of 3,5-difluorobenzimidamide

[0601] [ka]

[0602] Scheme 14A

[0603] [ka]

[0604] To a solution of 3,5-difluorobenzonitrile (1) (5.0 g, 36.0 mmol, 1.0 eq) in MeOH (50 mL) was added MeONa (3.9 g, 71.9 mmol, 2.0 eq), and the mixture was stirred at 20 °C for 4 h. To the mixture was added NHCl (9 g, 71.9 mmol, 2.0 eq), and the mixture was stirred at 40 °C for 16 h. It was cooled to RT, filtered, and the filtrate was concentrated to give 3,5-difluorobenzimidamide (2) (7 g, crude). LC-MS 157.4 [M+H] + .

[0605] Synthesis of 2-(3,5-difluorophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0606] [ka]

[0607] Scheme 14B

[0608] [ka]

[0609] To a mixture of 3,5-difluorobenzimidamide 2 (7 g, 44.9 mmol, 1.0 eq) and methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate 3 (6.2 g, 35.9 mmol, 0.8 eq) in MeOH (50 mL) was added KCO (12.4 g, 89.7 mmol, 2.0 eq) under N. The mixture was stirred at 75 °C for 2 h. The reaction mixture was concentrated to give 2-(3,5-difluorophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one 4, which was used directly in the next step. LC-MS 281.1 [M+H] + .

[0610] Synthesis of 2-(3,5-difluorophenyl)-4-((2-methoxyethoxy)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine

[0611] [ka]

[0612] Scheme 14C

[0613] [ka]

[0614] To a mixture of 2-(3,5-difluorophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (4) (crude, 1.0 eq) in DMA (50 mL) was added 1-(chloromethoxy)-2-methoxyethane (11.2 g, 89.7 mmol, 2.0 eq). The reaction mixture was stirred at RT for 3 h, then poured into HO (300 mL) and extracted with EtOAc (2 × 100 mL). The combined organic layers were washed with brine (300 mL), dried over NaSO, filtered, and concentrated. The residue was purified by column chromatography on silica gel eluting with 0–30% EtOAc / hexane to give 2-(3,5-difluorophenyl)-4-((2-methoxyethoxy)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine (5) (6 g). LC-MS: 369.2 [M+H] + .

[0615] Synthesis of (2,6-difluoro-4-(4-((2-methoxyethoxy)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)boronic acid

[0616] [ka]

[0617] Scheme 14D

[0618] [ka]

[0619] To a mixture of 2-(3,5-difluorophenyl)-4-((2-methoxyethoxy)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine (5) (400 mg, 1.4 mmol, 1.0 eq) in dry THF (10 mL) cooled to −78 °C, LDA (2 M, 1.1 mL, 1.5 eq) was added dropwise, and the mixture was stirred at −78 °C for 1 h. Trimethyl borate was added (191 mg, 1.9 mmol, 1.3 eq), and then the mixture was stirred at −78 °C for 3 h. The mixture was quenched with NH4Cl solution (30 mL) and extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by reverse-phase chromatography (0 to 50% acetonitrile / 0.05% formic acid) to give (2,6-difluoro-4-(4-((2-methoxyethoxy)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)boronic acid (6) (200 mg). LC-MS: 413.4 [M+H] + .

[0620] Scheme 14E

[0621] [ka]

[0622] A mixture of (2,6-difluoro-4-(4-((2-methoxyethoxy)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)boronic acid (6) (200 mg, 0.48 mmol, 1.0 eq) in FA (3 mL) was stirred at RT for 2 h. It was then concentrated and purified by Prep-HPLC to give (2,6-difluoro-4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)boronic acid (compound 14). LC-MS 325.1 [M+H] + . 1H NMR (400MHz, DMSO-d6): δ 12.85 (s, 1H), 8.86 (s, 2H), 7.72 (d, J = 7.2 Hz, 2H), 3.54 (s, 2H), 2.90 (s, 4H).

[0623] compound 15 Synthesis of 2-(3,5-difluoro-4-(2-(2-hydroxyethoxy)propan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (Compound 15)

[0624] [ka]

[0625] Synthesis of methyl 4-cyano-2,6-difluorobenzoate

[0626] [ka]

[0627] Scheme 15A

[0628] [ka]

[0629] To a solution of 4-cyano-2,6-difluorobenzoic acid (1) (5.00 g, 27.31 mmol, 1.0 eq) in THF (50 mL) / MeOH (50 mL) was added TMSCHN (2 M, 20.48 mL, 1.5 eq) dropwise at 25 °C. The mixture was stirred at 25 °C for 13 h. TLC (petroleum ether / ethyl acetate = 5 / 1, R f Cpd.1 = 0.33) indicated that 1 was completely consumed. The reaction mixture was concentrated under reduced pressure to give a residue. Methyl 4-cyano-2,6-difluorobenzoate 2 (4.1 g, crude) was obtained.

[0630] Synthesis of methyl 4-carbamimidoyl-2,6-difluorobenzoate

[0631] [ka]

[0632] Scheme 15B

[0633] [ka]

[0634] To a solution of methyl 4-cyano-2,6-difluorobenzoate (2) (2.00 g, 10.15 mmol, 1.0 eq) in THF (20 mL) was added LiHMDS (1 M, 15.22 mL, 1.5 eq) at 0 °C. The mixture was stirred at 20 °C for 16 h. LC-MS (ET68120-10-P1A1) showed that (2) was completely consumed, and the desired mass was detected. The reaction mixture was quenched by adding HCl / dioxane (4 mol / L, 8 mL) at 0 °C and then concentrated under reduced pressure to give a residue. The residue was triturated with MeOH (40 mL), and the mixture was stirred at 20 °C for 1 h. The mixture was then filtered, and the filter cake was dried under reduced pressure. Methyl 4-carbamimidoyl-2,6-difluorobenzoate (3) (2.6 g, crude, HCl) was used directly in the next step without further purification.

[0635] Synthesis of methyl 2,6-difluoro-4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)benzoate

[0636] [ka]

[0637] Scheme 15C

[0638] [ka]

[0639] To a solution of methyl 4-carbamimidoyl-2,6-difluorobenzoate (3) (470 mg, 1.88 mmol, 1.0 eq, HCl) and methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (3a) (327 mg, 1.88 mmol, 1.0 eq) in MeOH (4.7 mL) was added KCO (778 mg, 5.63 mmol, 3.0 eq) at 25 °C. The mixture was stirred at 25 °C for 16 h. LC-MS showed that 3 was completely consumed, with one peak corresponding to the desired mass. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (neutral conditions; column: Waters Xbridge Prep OBD C18 150 × 40 mm × 10 μm; mobile phase: [water (NH4HCO3)-ACN]; B%: 10% to 40%, 8 min) to give methyl 2,6-difluoro-4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)benzoate (4) (180 mg).

[0640] Synthesis of 2-(3,5-difluoro-4-(2-hydroxypropan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0641] [ka]

[0642] Scheme 15D

[0643] [ka]

[0644] To a solution of methyl 2,6-difluoro-4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)benzoate (4) (300 mg, 887 μmol, 1.0 eq) in THF (5.0 mL) was added MeMgBr (3 M, 1.03 mL, 3.5 eq) at 0 °C. The mixture was stirred at 25 °C for 1 h. TLC (petroleum ether / ethyl acetate = 1 / 1, R f Cpd.4 = 0.60) indicated that 4 was completely consumed, and two new spots were formed. LC-MS (ET65158-9-P1A1) showed that 4 was completely consumed, with one main peak of the desired MS. The reaction mixture was quenched by adding NH4Cl (15 mL) at 0 °C and then extracted with 45 mL of EtOAc (15 mL × 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (petroleum ether / ethyl acetate = 1 / 1). 2-(3,5-difluoro-4-(2-hydroxypropan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (150 mg, crude) was obtained, which was used immediately in the next step. LC-MS (ET65158-9-P1A1, Product: R t = 0.575 mins).

[0645] Scheme 15E

[0646] [ka]

[0647] To a solution of 2-(3,5-difluoro-4-(2-hydroxypropan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (20.0 mg, 59.11 μmol, 1.0 eq) in ethylene glycol (1.59 g, 25.62 mmol, 1.43 mL, 433.41 eq), TosOH (12.7 mg, 73.88 μmol, 1.25 eq) was added at 25 °C. The mixture was stirred at 80 °C for 12 h. TLC (petroleum ether / ethyl acetate = 0 / 1, R f The Cpd.5 (Cpd.5) = 0.53) indicated the complete consumption of 5, and two new spots formed. After cooling to 25 °C, the mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO2, petroleum ether / ethyl acetate = 0 / 1) to give 2-(3,5-difluoro-4-(2-(2-hydroxyethoxy)propan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 15). LC-MS: 383.1 (M+1). 1 H NMR (400 MHz, DMSO-d6): δ= 7.78 - 7.70 (m, 2H), 4.49 - 4.45 (m, 1H) 3.47 - 3.43 (m, 4H), 3.21 - 3.17 (m, 2H), 2.85~2.75 (m, 4H), 1.637 (s, 6H).

[0648] compound 16 Synthesis of 2-(3,5-difluoro-4-(2-(2-methoxyethoxy)propan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (Compound 16)

[0649] [ka]

[0650] Scheme 16A

[0651] [ka]

[0652] The synthesis of 2-(3,5-difluoro-4-(2-hydroxypropan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) is shown in Schemes 15A-15D.

[0653] To a solution of 2-(3,5-difluoro-4-(2-hydroxypropan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (50.0 mg, 147 μmol, 1.0 eq) in 2-methoxyethanol (0.5 mL) was added TosOH (31.6 mg, 183 μmol, 1.24 eq) at 25 °C, then heated to 85 °C and stirred for 13 h. LC-MS showed that ∼17% of (5) remained. Several new peaks were detected by LC-MS, indicating the desired compound. After cooling to 25 °C, the reaction mixture was purified by prep-HPLC (neutral conditions; column: Waters Xbridge BEH C18 100 × 30 mm × 10 μm; mobile phase: [water (NH4HCO3))-can]; B%: 25% to 55%, 8 min) to give 2-(3,5-difluoro-4-(2-(2-methoxyethoxy)propan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 16). LC-MS: 397.1 (M+1). 1 H NMR (400 MHz, DMSO-d6): δ= 7.81~7.75 (m, 2H), 3.549 (s, 2H) 3.53~3.38 (m, 2H), 3.35~3.25 (m, 2H), 3.22 (s, 3H), 2.906 (m, 4H), 1.653 (s, 6H).

[0654] compound 17 Synthesis of 2-(4-(2-methoxyethoxy)propan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (Compound 17)

[0655] [ka]

[0656] Synthesis of 1-bromo-4-(2-(2-methoxyethoxy)propan-2-yl)benzene

[0657] [ka]

[0658] Scheme 17A

[0659] [ka]

[0660] To a solution of 2-(4-bromophenyl)propan-2-ol (1) (2.6 g, 12.2 mmol, 1.0 eq) in 2-methoxyethan-1-ol (20 mL) was added TsOH·HO (2.3 g, 12.2 mmol, 1 eq). The mixture was stirred at RT for 2 days, then quenched with water (100 mL) and extracted with EtOAc (3 × 60 mL). The organic layer was concentrated, and the residue was purified by column chromatography on silica gel eluting with 0–30% EtOAc / hexane to give 1-bromo-4-(2-(2-methoxyethoxy)propan-2-yl)benzene (2) (2.7 g). 1 H NMR (400MHz, DMSO-d6): δ 7.54-7.51 (m, 2H), 7.38-7.35 (m, 2H), 3.42-3.40 (m, 2H), 3.24-3.22 (m, 5H), 1.44 (s, 6H).

[0661] Synthesis of 4-(2-(2-methoxyethoxy)propan-2-yl)benzonitrile

[0662] [ka]

[0663] Scheme 17B

[0664] [ka]

[0665] To a solution of 1-bromo-4-(2-(2-methoxyethoxy)propan-2-yl)benzene (2) (1.5 g, 3.7 mmol, 1 eq) in NMP (10 mL) was added Zn(CN) (260 mg, 2.2 mmol, 0.6 eq) and Pd(PPh) (430 mg, 0.37 mmol, 0.1 eq). The resulting solution was stirred at 140 °C under Ar for 2 h. The resulting mixture was cooled to RT, diluted with water (50 mL), and extracted with EtOAc (3 × 60 mL). The organic layers were combined, washed with brine (100 mL), dried over anhydrous sodium sulfate, and concentrated. The residue was purified by column chromatography on silica gel eluting with 0–40% EtOAc / hexane to give 4-(2-(2-methoxyethoxy)propan-2-yl)benzonitrile (3) (600 mg). 1 H NMR (400MHz, DMSO-d6): δ 7.81 (d, J = 8.4 Hz, 2H), 7.62 (d, J = 8.4 Hz, 2H), 3.44 (t, J = 5.2 Hz, 2H), 3.28-3.25 (m, 5H), 1.47 (s, 6H).

[0666] Synthesis of 4-(2-(2-methoxyethoxy)propan-2-yl)benzimidamide

[0667] [ka]

[0668] Scheme 17C

[0669] [ka]

[0670] To a solution of 4-(2-(2-methoxyethoxy)propan-2-yl)benzonitrile (3) (0.6 g, 2.7 mmol, 1.0 eq) in MeOH (5 mL) was added MeONa (295 mg, 5.5 mmol, 2.0 eq), and the mixture was stirred at RT for 4 h. NHCl (287 mg, 5.5 mmol, 2.0 eq) was added, and the mixture was stirred at 40 °C for 16 h. The mixture was cooled to RT, filtered, and the filtrate was concentrated to give 4-(2-(2-methoxyethoxy)propan-2-yl)benzimidamide (4) (285 mg, crude). LC-MS: 237.2 [M+H] + .

[0671] Scheme 17D

[0672] [ka]

[0673] To a solution of 4-(2-(2-methoxyethoxy)propan-2-yl)benzimidamide (4) (150 mg, 0.7 mmol, 1.0 eq) in MeOH (5 mL) was added KCO (180 mg, 1.3 mmol, 2.0 eq) and methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (80 mg, 0.5 mmol, 0.7 eq) under N. The mixture was stirred at 70 °C for 2 h. It was then concentrated and purified by prep-HPLC to give 2-(4-(2-methoxyethoxy)propan-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 17). LC-MS: 361.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ 12.70 (s, 1H), 8.06 (d, J = 8.6 Hz, 2H), 7.54 (d, J = 8.6 Hz, 2H), 3.52 (s, 2H), 3.46-3.41 (m, 2H), 3.25 (d, J = 5.0 Hz, 5H), 2.88 (dd, J = 7.6, 4.0 Hz, 4H), 1.48 (s, 6H).

[0674] compound 18 Synthesis of methyl (2-((2-(4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)propan-2-yl)oxy)ethyl)carbamate (Compound 18)

[0675] [ka]

[0676] Synthesis of 1-bromo-4-(2-(2-nitroethoxy)propan-2-yl)benzene

[0677] [ka]

[0678] Scheme 18A

[0679] [ka]

[0680] To a solution of 2-(4-bromophenyl)propan-2-ol 1 (2.6 g, 12.2 mmol, 1.0 eq) in DCM (20 mL) was added TFA (2.3 g, 12.2 mmol, 1.0 eq) and 2-nitroethan-1-ol 2 (10 mL). The mixture was stirred at RT for 3 days. The reaction was quenched with water (100 mL) and extracted with EtOAc (3 × 60 mL). The organic layer was concentrated, and the residue was purified by column chromatography on silica gel eluting with 0–30% EtOAc / hexane to give 1-bromo-4-(2-(2-nitroethoxy)propan-2-yl)benzene 3 (2.7 g). 1 H NMR (400 MHz, CDCl3): δ 7.80 (d, J = 8.8 Hz, 2H), 7.31 (d, J = 8.4 Hz, 2H), 4.55 (t, J = 4.8 Hz, 2H), 3.69 (t, J = 4.8 Hz, 2H), 1.51 (s, 6H).

[0681] Synthesis of 2-((2-(4-bromophenyl)propan-2-yl)oxy)ethan-1-amine

[0682] [ka]

[0683] Scheme 18B

[0684] [ka]

[0685] To a solution of 1-bromo-4-(2-(2-nitroethoxy)propan-2-yl)benzene 3 (1.5 g, 3.7 mmol, 1.0 eq) in EtOH / HO (10 mL / 2 mL) was added Zn (1.2 g, 18.5 mmol, 5.0 eq) and NHCl (1.0 g, 18.5 mmol, 5.0 eq). The resulting mixture was stirred at 80 °C for 12 h under Ar. The reaction mixture was cooled to RT, diluted with water (50 mL), and extracted with EtOAc (3 × 60 mL). The organic layers were combined, washed with brine (100 mL), dried over anhydrous sodium sulfate, and concentrated to give 2-((2-(4-bromophenyl)propan-2-yl)oxy)ethan-1-amine 4 (1.2 g, crude). LC-MS: 258.0, 260.0 [M+H] + .

[0686] Synthesis of methyl (2-((2-(4-bromophenyl)propan-2-yl)oxy)ethyl)carbamate

[0687] [ka]

[0688] Scheme 18C

[0689] [ka]

[0690] To a solution of 2-((2-(4-bromophenyl)propan-2-yl)oxy)ethan-1-amine (4) (1.2 g, 4.7 mmol, 1.0 eq) in DCM (10 mL) was added TEA (707 mg, 7.0 mmol, 1.5 eq), followed by methyl carbonochloridate (5) (395 mg, 4.2 mmol, 0.9 eq). The mixture was stirred at RT for 1 day. The reaction was concentrated, and the residue was purified by column chromatography on silica gel eluting with 0–30% EtOAc / hexane to give methyl (2-((2-(4-bromophenyl)propan-2-yl)oxy)ethyl)carbamate (6) (1.0 g). LC-MS: 357.0, 359.0 [M+H+MeCN] + .

[0691] Synthesis of methyl (2-((2-(4-cyanophenyl)propan-2-yl)oxy)ethyl)carbamate

[0692] [ka]

[0693] Scheme 18D

[0694] [ka]

[0695] To a solution of methyl (2-((2-(4-bromophenyl)propan-2-yl)oxy)ethyl)carbamate (6) (1.0 g, 3.7 mmol, 1 eq) in NMP (10 mL) was added Zn(CN) (260 mg, 2.2 mmol, 0.6 eq), followed by Pd(PPh) (430 mg, 0.37 mmol, 0.1 eq). The resulting solution was stirred at 140 °C under Ar for 2 h. The resulting mixture was cooled to RT, diluted with water (50 mL), and extracted with EtOAc (3 × 20 mL). The organic layers were combined, washed with brine (100 mL), dried over anhydrous sodium sulfate, and concentrated. The residue was purified by column chromatography on silica gel eluted with 0-40% EtOAc / hexane to give methyl (2-((2-(4-cyanophenyl)propan-2-yl)oxy)ethyl)carbamate (7) (600 mg). 1 H NMR (400 MHz, DMSO): δ 7.86 - 7.76 (m, 2H), 7.64 - 7.58 (m, 2H), 7.25 - 7.11 (m, 1H), 3.33 (s, 3H), 3.15 - 3.04 (m, 4H), 1.46 (s, 6H).

[0696] Synthesis of methyl (2-((2-(4-carbamimidoylphenyl)propan-2-yl)oxy)ethyl)carbamate

[0697] [ka]

[0698] Scheme 18E

[0699] [ka]

[0700] To a solution of methyl (2-((2-(4-cyanophenyl)propan-2-yl)oxy)ethyl)carbamate 7 (0.6 g, 2.7 mmol, 1.0 eq) in MeOH (5 mL) was added MeONa (295 mg, 5.5 mmol, 2.0 eq) and the mixture was stirred at RT for 4 h. NHCl (287 mg, 5.5 mmol, 2.0 eq) was added and the mixture was stirred at 40 °C for 16 h. The mixture was cooled to RT, filtered, and the filtrate was concentrated to give methyl (2-((2-(4-carbamimidoylphenyl)propan-2-yl)oxy)ethyl)carbamate 8 (285 mg, crude). LC-MS: 280.2 [M+H] + .

[0701] Scheme 18F

[0702] [ka]

[0703] To a solution of methyl (2-((2-(4-carbamimidoylphenyl)propan-2-yl)oxy)ethyl)carbamate (8) (285 mg, 0.7 mmol, 1.0 eq) in MeOH (5 mL) was added KCO (180 mg, 1.3 mmol, 2.0 eq) and methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (80 mg, 0.5 mmol, 0.7 eq) under N. The mixture was stirred at 70 °C for 2 h, then concentrated and purified by prep-HPLC to give methyl (2-((2-(4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)propan-2-yl)oxy)ethyl)carbamate (compound 18). LC-MS: 404.2 [M+H] + . 1H NMR (400 MHz, DMSO): δ 12.71 (s, 1H), 8.05 (d, J = 8.2 Hz, 2H), 7.56 (d, J = 8.4 Hz, 2H), 7.16 (s, 1H), 3.51 (m, 5H), 3.12 (dd, J = 12.5, 5.0 Hz, 4H), 2.89 (d, J = 4.0 Hz, 4H), 1.47 (s, 6H).

[0704] compound 19 Synthesis of 2-(6'-bromo-[2,3'-bipyridin]-5-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (Compound 19)

[0705] [ka]

[0706] Synthesis of 6'-bromo-[2,3'-bipyridine]-5-carbonitrile

[0707] [ka]

[0708] Scheme 19A

[0709] [ka]

[0710] To a solution of 6-bromonicotinonitrile (1) (2.00 g, 10.93 mmol, 1.0 eq) and (6-bromopyridin-3-yl)boronic acid (1a) (2.00 g, 9.91 mmol, 9.07 e-1 eq) in dioxane (20 mL) / HO (4 mL) was added KCO (3.02 g, 21.86 mmol, 2.0 eq) and Pd(dppf)Cl (799.65 mg, 1.09 mmol, 0.1 eq) at 20 °C under N. The mixture was stirred at 100 °C for 16 h under N. After cooling to 25 °C, the mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue, which was purified by column chromatography (SiO, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1). 6'-Bromo-[2,3'-bipyridine]-5-carbonitrile (2) (730 mg) was obtained. 1 H NMR: (400 MHz, CDCl3): δ= 8.92 (dd, J = 1.8, 12.3 Hz, 2H), 8.19 (dd, J = 2.5, 8.3 Hz, 1H), 8.00 (dd, J = 2.0, 8.3 Hz, 1H), 7.81 (d, J = 8.3 Hz, 1H), 7.58 (d, J = 8.3 Hz, 1H)

[0711] Synthesis of 6'-bromo-[2,3'-bipyridine]-5-carboximidamide

[0712] [ka]

[0713] Scheme 19B

[0714] [ka]

[0715] To a solution of 6'-bromo-[2,3'-bipyridine]-5-carbonitrile (2) (280 mg, 1.08 mmol, 1.0 eq) in THF (3 mL) was added LiHMDS (1 M, 2.69 mL, 2.5 eq) under N at 0 °C. The mixture was warmed to 20 °C and stirred at 20 °C for 12 h. The reaction mixture was quenched by the addition of HCl / dioxane (4 mol / L, 1 mL) at 0 °C and then concentrated under reduced pressure to give a residue. The residue was suspended in MeOH (7 mL), and the mixture was stirred at 20 °C for 1 h. The mixture was then filtered, and the filter cake was dried under reduced pressure to give 6'-bromo-[2,3'-bipyridine]-5-carboximidamide (3) (350 mg, crude, HCl), which was used immediately in the next step.

[0716] Scheme 19C

[0717] [ka]

[0718] To a solution of 6'-bromo-[2,3'-bipyridine]-5-carboximidamide (3) (350 mg, 1.12 mmol, 1.0 eq, HCl) in MeOH (3.5 mL) was added methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (3a) (194.45 mg, 1.12 mmol, 1.0 eq) and K2CO3 (462.79 mg, 3.35 mmol, 3.0 eq) at 20 °C. The mixture was stirred at 60 °C for 16 h. The mixture was filtered and concentrated under reduced pressure to give a residue, which was purified by prep-HPLC (neutral conditions; column: Phenomenex C18 75 × 30 mm × 3 μm; mobile phase: [water (NH4HCO3)-ACN]; B%: 25% to 40%, 10 min). 2-(6'-Bromo-[2,3'-bipyridin]-5-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 19) was obtained. LC-MS: 401 (M+1). 1H NMR: (400 MHz, DMSO-d6): δ= 9.27 (d, J = 1.3 Hz, 1H), 8.94 (d, J = 2.2 Hz, 1H), 8.43 (dd, J = 1.9, 8.3 Hz, 1H), 8.27 (dd, J = 2.3, 8.4 Hz, 1H), 7.90 (d, J = 8.3 Hz, 1H), 7.61 (d, J = 8.4 Hz, 1H), 3.28 (s, 2H), 2.61 (br dd, J = 5.0, 15.4 Hz, 4H).

[0719] compound 20 Synthesis of 2-(4-(6-bromopyridin-3-yl)-3-fluorophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 20)

[0720] [ka]

[0721] Synthesis of 4-(6-bromopyridin-3-yl)-3-fluorobenzonitrile

[0722] [ka]

[0723] Scheme 20A

[0724] [ka]

[0725] To a solution of (6-bromopyridin-3-yl)boronic acid (1a) (408.52 mg, 2.02 mmol, 1.0 eq) in dioxane (5 mL) / HO (1 mL) was added 3-fluoro-4-iodobenzonitrile (1) (0.5 g, 2.02 mmol, 1.0 eq), KCO (559.52 mg, 4.05 mmol, 2.0 eq), and Pd(dppf)Cl (148.11 mg, 202.42 μmol, 0.1 eq) at 20 °C under N. The mixture was stirred at 100 °C for 16 h under N. After cooling to 20 °C, the mixture was filtered and concentrated under reduced pressure to give a residue, which was purified by column chromatography (SiO, petroleum ether / ethyl acetate = 20 / 1 to 0 / 1). 4-(6-bromopyridin-3-yl)-3-fluorobenzonitrile (2) (200 mg, crude) was obtained, which was used directly in the next step. 1 H NMR: (400 MHz, CDCl3): δ= 8.48 (s, 1H), 7.68 (td, J = 1.9, 8.3 Hz, 1H), 7.56 (s, 1H), 7.50 (br d, J = 7.5 Hz, 2H), 7.47 - 7.45 (m, 1H).

[0726] Synthesis of 4-(6-bromopyridin-3-yl)-3-fluorobenzimidamide

[0727] [ka]

[0728] Scheme 20B

[0729] [ka]

[0730] To a solution of 4-(6-bromopyridin-3-yl)-3-fluorobenzonitrile 2 (200 mg, 721.78 μmol, 1.0 eq) in THF (2 mL) was added LiHMDS (1 M, 1.80 mL, 2.5 eq) under N at 0 °C. The mixture was warmed to 20 °C and stirred at 20 °C for 16 h. The reaction mixture was quenched by the addition of HCl / dioxane (4 mol / L, 0.6 mL) at 0 °C and then concentrated under reduced pressure to give a residue. This residue was suspended in MeOH (3 mL), and the mixture was stirred at 20 °C for 1 h. The mixture was then filtered, and the filter cake was dried under reduced pressure to give 4-(6-bromopyridin-3-yl)-3-fluorobenzimidamide 3 (0.11 g, crude), which was used in the next step without further purification.

[0731] Scheme 20C

[0732] [ka]

[0733] To a solution of methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (3a) (68.75 mg, 394.62 μmol, 1.06 eq) and 4-(6-bromopyridin-3-yl)-3-fluorobenzimidamide (3) (0.11 g, 373.99 μmol, 1.0 eq) in MeOH (2 mL) was added KCO (183.33 mg, 1.33 mmol, 3.55 eq) at 20° C. The mixture was stirred at 60° C. for 16 h. After cooling to 20 °C, the mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. This residue was purified by prep-HPLC (neutral conditions; column: Waters Xbridge Prep OBD C18 150 × 40 mm × 10 μm; mobile phase: [water (NH3H2O ​​+ NH4HCO3)-ACN]; B%: 30% to 60%, 8 min) to give 2-(4-(6-bromopyridin-3-yl)-3-fluorophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 20). LC-MS: 417 + bromo isomer (M+1). 1 H NMR: (400 MHz, DMSO-d6): δ 8.66 (s, 1H), 8.13 - 8.06 (m, 2H), 8.06 - 8.00 (m, 1H), 7.86 - 7.77 (m, 2H), 3.55 (s, 2H), 2.91 (s, 4H).

[0734] compound 21 Synthesis of 2-(4-(6-bromopyridin-3-yl)-3,5-difluorophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (Compound 21)

[0735] [ka]

[0736] Synthesis of 3,5-difluoro-4-iodobenzonitrile

[0737] [ka]

[0738] Scheme 21A

[0739] [ka]

[0740] To a solution of 3,5-difluorobenzonitrile (1) (2.00 g, 14.07 mmol, 1.0 eq) in THF (20 mL) was added LDA (2 M, 7.86 mL, 1.12 eq) dropwise at −70° C. Then, I (3.76 g, 14.80 mmol, 2.98 mL, 1.05 eq) in THF (10 mL) was added at −70° C. The reaction mixture was slowly warmed to 15° C. and then stirred at 15° C. for 1 h. The mixture was quenched with 10% sodium thiosulfite solution (15 mL). The reaction mixture was extracted with a 1 / 1 mixture of ethyl acetate / hexane (7 mL × 3). The combined organic phases were dried over NaSO and concentrated in vacuo. The residue was purified by prep-HPLC (neutral conditions; column: Welch Xtimate C18 250 × 70 mm #10 μm; mobile phase: [water (NH4HCO3)-ACN]; B%: 40% to 70%, 20 min) to give 3,5-difluoro-4-iodobenzonitrile (2) (2.00 g). 1 H NMR: (400 MHz, CDCl3): δ= 7.23 - 7.18 (m, 2H).

[0741] Synthesis of 4-(6-bromopyridin-3-yl)-3,5-difluorobenzonitrile

[0742] [ka]

[0743] Scheme 21B

[0744] [ka]

[0745] To a solution of (6-bromopyridin-3-yl)boronic acid (2a) (1.60 g, 7.93 mmol, 1.05 eq) and 3,5-difluoro-4-iodobenzonitrile (2) (2.00 g, 11.32 mmol, 1.0 eq) in dioxane (20 mL) / HO (4 mL) was added KCO (2.09 g, 15.09 mmol, 2.0 eq) and Pd(dppf)Cl (552.23 mg, 0.75 mmol, 0.1 eq) at 15 °C under N. The mixture was heated to 100 °C for 16 h under N. After cooling to 15 °C, the mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1). 4-(6-Bromopyridin-3-yl)-3,5-difluorobenzonitrile (3) (620 mg, 210 mmol) was obtained. 1 H NMR: (400 MHz, CDCl3): δ= 8.43 (s, 1H), 7.63 - 7.55 (m, 2H), 7.34 - 7.26 (m, 2H).

[0746] Synthesis of 4-(6-bromopyridin-3-yl)-3,5-difluorobenzimidamide

[0747] [ka]

[0748] Scheme 21C

[0749] [ka]

[0750] To a solution of 4-(6-bromopyridin-3-yl)-3,5-difluorobenzonitrile (3) (500 mg, 1.69 mmol, 1.0 eq) in THF (5 mL) was added LiHMDS (1 M, 4.24 mL, 2.5 eq) under N2 at 0 °C. The reaction mixture was quenched by adding HCl / dioxane (4 mol / L, 2.0 mL) at 0 °C and then concentrated under reduced pressure to obtain a residue. The residue was suspended in MeOH (8 ml), and the mixture was stirred at 20 °C for 1 hour. The mixture was then filtered, and the filtrate was concentrated under reduced pressure to obtain 4-(6-bromopyridin-3-yl)-3,5-difluorobenzimidamide (4) (540 mg, crude, HCl). This compound was used immediately in the next step.

[0751] Scheme 21D

[0752] [ka]

[0753] To a solution of 4-(6-bromopyridin-3-yl)-3,5-difluorobenzimidamide (4) (440 mg, 1.41 mmol, 1.0 eq) in MeOH (4.4 mL), methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (4a) (245.60 mg, 1.41 mmol, 1.0 eq) and KCO (584.52 mg, 4.23 mmol, 3.0 eq) were added at 20 °C. The mixture was stirred at 20 °C for 16 h. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (neutral conditions; column: NP-1; mobile phase: [heptane-EtOH]; B%: 10% to 70%, 10 min). 2-(4-(6-Bromopyridin-3-yl)-3,5-difluorophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 21) was obtained. LC-MS: 438 + bromo isomer (M+1). 1H NMR: (400 MHz, DMSO-d6): δ= 13.06 - 12.89 (m, 1H), 8.64 (s, 1H), 8.10 - 8.00 (m, 3H), 7.93 (d, J = 8.4 Hz, 1H), 3.62 (s, 2H), 2.98 (s, 4H).

[0754] compound 22 Synthesis of 6-(4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)pyridin-3-yl)boronic acid (compound 22)

[0755] [ka]

[0756] Synthesis of 4-(5-bromopyridin-2-yl)benzonitrile

[0757] [ka]

[0758] Scheme 22A

[0759] [ka]

[0760] To a solution of 5-bromo-2-iodopyridine (1) (0.5 g, 1.76 mmol, 1.0 eq) and (4-cyanophenyl)boronic acid (1a) (388.19 mg, 2.64 mmol, 1.5 eq) in dioxane (5 mL) / HO (1 mL), Pd(dppf)Cl (128.87 mg, 176.12 μmol, 0.1 eq) and KCO (486.83 mg, 3.52 mmol, 2.0 eq) were added under N at 20 °C. The mixture was stirred at 100 °C for 13 h. After cooling to 20 °C, the mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1). 4-(5-bromopyridin-2-yl)benzonitrile (2) (400 mg, crude) was obtained, which was used immediately in the next step. 1 H NMR: (400 MHz, CDCl3): δ= 8.71 (d, J = 2.1 Hz, 1H), 8.06 - 8.00 (m, 2H), 7.87.

[0761] Synthesis of 4-(5-bromopyridin-2-yl)benzimidamide

[0762] [ka]

[0763] Scheme 22B

[0764] [ka]

[0765] To a solution of 4-(5-bromopyridin-2-yl)benzonitrile 2 (100 mg, 385.95 μmol, 1.0 eq) in THF (1 mL) was added LiHMDS (1 M, 964.87 μL, 2.5 eq) under N at 0 °C. The mixture was warmed to 20 °C and stirred for 16 h. The reaction mixture was quenched by the addition of HCl / dioxane (4 mol / L, 0.3 mL) at 0 °C and then concentrated under reduced pressure to give a residue. The residue was suspended in MeOH (2 ml), and the mixture was stirred at 20 °C for 1 h. The mixture was then filtered, and the filtrate was concentrated under reduced pressure to give a residue. 4-(5-bromopyridin-2-yl)benzimidamide 3 (110 mg, crude, HCl salt) was obtained, which was used in the next step without further purification.

[0766] Synthesis of 2-(4-(5-bromopyridin-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0767] [ka]

[0768] Scheme 22C

[0769] [ka]

[0770] To a solution of 4-(5-bromopyridin-2-yl)benzimidamide 3 (100 mg, 362.15 μmol, 1.0 eq) in MeOH (1 mL) was added methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate 3a (126.18 mg, 724.29 μmol, 2.0 eq) and KCO (100.10 mg, 724.29 μmol, 2.0 eq) at 20 °C. The mixture was stirred at 60 °C for 16 h. The mixture was filtered, and the filter cake was dried under reduced pressure to give 2-(4-(5-bromopyridin-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one 4 (238 mg, crude), which was used immediately in the next step. 1 H NMR: (400 MHz, CDCl3): δ= 8.75 - 8.69 (m, 1H), 8.14 - 8.04 (m, 4H), 7.90 - 7.83 (m, 1H), 7.68 - 7.61 (m, 1H), 3.65 - 3.62 (m, 2H), 3.04 - 2.97 (m, 2H), 2.92 - 2.86 (m, 2H).

[0771] Scheme 22D

[0772] [ka]

[0773] To a solution of 2-(4-(5-bromopyridin-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (4) (238 mg, 594.57 μmol, 1.0 eq) in dioxane (3 mL), B2Pin2 (181.18 mg, 713.48 μmol, 1.2 eq), KOAc (175.06 mg, 1.78 mmol, 3.0 eq), and Pd(dppf)Cl.DCM (48.55 mg, 59.46 μmol, 0.1 eq) were added under N2. The mixture was stirred at 80 °C for 13 h. After cooling to 25 °C, the mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (neutral conditions; column: Waters Xbridge Prep OBD C18 150 × 40 mm × 10 μm; mobile phase: [water (NH4HCO3)-ACN]; B%: 15% to 45%, 8 min) to give (6-(4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)pyridin-3-yl)boronic acid (compound 22). LC-MS: 366.1 (M+1). 1 H NMR: (400 MHz, DMSO-d6): δ= 12.83 (br s, 1H), 9.01 (s, 1H), 8.41 (s, 2H), 8.30 - 8.20 (m, 5H), 8.06 (d, J = 8.0 Hz, 1H), 3.55 (s, 2H), 2.92 (s, 4H).

[0774] compound 23 Synthesis of 2-(4-(2-bromopyridin-4-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 23)

[0775] [ka]

[0776] Synthesis of 4-(2-bromopyridin-4-yl)benzonitrile

[0777] [ka]

[0778] Scheme 23A

[0779] [ka]

[0780] To a solution of 2-bromo-4-iodopyridine 1 (2.50 g, 8.81 mmol, 1.00 eq) and (4-cyanophenyl)boronic acid 1a (1.42 g, 9.69 mmol, 1.10 eq) in dioxane (25.0 mL) and HO (5.00 mL) was added KCO (2.43 g, 17.6 mmol, 2.00 eq) at 20 °C. The suspension was degassed and purged with N three times. Pd(dppf)Cl (644 mg, 881 μmol, 0.10 eq) was then added to the mixture under N at 20 °C. The suspension was degassed and purged with N three times. The reaction mixture was stirred at 100 °C for 12 h under N. The reaction mixture was diluted with H2O (10.0 mL) and extracted with 60.0 mL of EtOAc (20.0 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 1 / 1). 4-(2-bromopyridin-4-yl)benzonitrile (2) (950 mg) was obtained. 1 H NMR: (400 MHz, DMSO-d6): δ= 8.57 (d, J = 5.3 Hz, 1H), 8.17 - 8.03 (m, 5H), 7.91 (dd, J = 1.7, 5.2 Hz, 1H).

[0781] Synthesis of 4-(2-bromopyridin-4-yl)benzimidamide

[0782] [ka]

[0783] Scheme 23B

[0784] [ka]

[0785] To a solution of 4-(2-bromopyridin-4-yl)benzonitrile 2 (400 mg, 1.54 mmol, 1.00 eq) in THF (4.00 mL) was added LiHMDS (1 M, 3.09 mL, 2.00 eq) dropwise at 0 °C. The mixture was stirred at 20 °C for 16 h. The reaction mixture was diluted with 4 N HCl (4.00 mL). The combined organic layers were concentrated under reduced pressure to give a residue. The residue was suspended in MeOH (20.0 mL) and stirred at 20 °C for 1 h. The mixture was then filtered. The filtrate was concentrated under reduced pressure to give a residue. 4-(2-bromopyridin-4-yl)benzimidamide 3 (500 mg, crude) was obtained, which was used in the next step without further purification.

[0786] Scheme 23C

[0787] [ka]

[0788] To a solution of 4-(2-bromopyridin-4-yl)benzimidamide (3) (250 mg, 905 μmol, 1.00 eq) and methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (3a) (166 mg, 951 μmol, 1.05 eq) in MeOH (2.50 mL) was added KCO (313 mg, 2.26 mmol, 2.50 eq) at 20 °C. The mixture was stirred at 60 °C for 16 h. The reaction mixture was filtered. The filter cake was slurried with MeOH (0.50 mL) and HO (1.00 mL) and then filtered. The filter cake was dried in vacuo to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 80 × 40 mm × 3 μm; mobile phase: [water (HCl)-ACN]; B%: 35% to 65%, 7 min) to give 2-(4-(2-bromopyridin-4-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (compound 23). 1 H NMR (400 MHz, DMSO-d6): δ= 8.48 (d, J = 5.3 Hz, 1H), 8.27 (d, J = 8.4 Hz, 2H), 8.07 (s, 1H), 7.99 (d, J = 8.4 Hz, 2H), 7.87 (dd, J = 1.2, 5.2 Hz, 1H), 3.54 (s, 2H), 2.89 (s, 4H). LC-MS: 404.1 (M+1).

[0789] compound 24 Synthesis of 2-(4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)pyrimidin-5-yl)boronic acid (compound 24)

[0790] [ka]

[0791] Synthesis of 4-(5-bromopyrimidin-2-yl)benzonitrile

[0792] [ka]

[0793] Scheme 24A

[0794] [ka]

[0795] To a solution of 5-bromo-2-iodopyrimidine (1) (2.50 g, 8.78 mmol, 1.00 eq) and (4-cyanophenyl)boronic acid (1a) (1.42 g, 9.65 mmol, 1.10 eq) in dioxane (25.0 mL) and HO (5.00 mL) was added KCO (2.43 g, 17.6 mmol, 2.00 eq) at 15 °C. The suspension was degassed and purged with N three times. To the mixture was then added Pd(dppf)Cl (642 mg, 878 μmol, 0.10 eq) under N at 15 °C. The suspension was degassed and purged with N three times. The mixture was stirred at 100 °C under N for 16 h. The reaction mixture was diluted with HO (10.0 mL) and filtered. The filtrate was extracted with EtOAc (10 mL × 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. The residue was suspended in MTBE (10.0 mL), stirred at 15 °C for 16 h, and then filtered. The filter cake was dried under reduced pressure to obtain a residue. 4-(5-bromopyrimidin-2-yl)benzonitrile (2) (2.00 g, crude) was obtained, which was immediately used in the next step. 1 H NMR (400 MHz, DMSO-d6): δ= 9.16 (s, 2H), 8.49 (br d, J = 8.3 Hz, 2H), 8.01 (br d, J = 8.3 Hz, 2H).

[0796] Synthesis of 4-(5-bromopyrimidin-2-yl)benzimidamide

[0797] [ka]

[0798] Scheme 24B

[0799] [ka]

[0800] To a solution of 4-(5-bromopyrimidin-2-yl)benzonitrile 2 (0.80 g, 3.08 mmol, 1.00 eq) in THF (8.00 mL) was added LiHMDS (1 M, 6.15 mL, 2.00 eq) at 0 °C. The mixture was stirred at 15 °C for 16 h. The reaction mixture was diluted with 4 N HCl / dioxane (6.00 mL). The combined organic layers were concentrated under reduced pressure to give a residue. The residue was suspended in MeOH (8.00 mL), stirred at 15 °C for 2 h, and then filtered. The filtrate was concentrated under reduced pressure to give a residue. 4-(5-bromopyrimidin-2-yl)benzimidamide 3 (0.8 g, crude) was obtained, which was used in the next step without further purification.

[0801] Synthesis of 2-(4-(5-bromopyrimidin-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0802] [ka]

[0803] Scheme 24C

[0804] [ka]

[0805] To a solution of 4-(5-bromopyrimidin-2-yl)benzimidamide (3) (0.80 g, 2.89 mmol, 1.00 eq) and methyl 4-oxotetrahydrothiopyran-3-carboxylate (6) (528 mg, 3.03 mmol, 1.05 eq) in MeOH (16.0 mL) was added KCO (1.20 g, 8.66 mmol, 3.00 eq) at 25 °C. The mixture was stirred at 60 °C for 16 h. The reaction mixture was filtered. The filtrate was concentrated under reduced pressure. The resulting solid was suspended in water (5 mL) and stirred at 20 °C for 3 h. The mixture was then filtered, and the filter cake was dried in vacuo to give 2-(4-(5-bromopyrimidin-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (4) (300 mg, crude). The crude product was used directly in the next step without further purification.

[0806] Scheme 24D

[0807] [ka]

[0808] To a solution of 2-(4-(5-bromopyrimidin-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (4) (0.1 g, 249.20 μmol, 1 eq) in dioxane (2 mL) was added 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (69.61 mg, 274.12 μmol, 1.1 eq), KOAc (48.91 mg, 498.41 μmol, 2 eq), and Pd(dppf)Cl.CHCl (20.35 mg, 24.92 μmol, 0.1 eq) under N at 20 °C. The mixture was stirred at 80 °C for 12 h under N. The mixture was concentrated under reduced pressure to give a residue. The residue was suspended in MeCN / HO (1:1, 3 mL) and stirred at 20 °C for 2 h. The mixture was then filtered, and the filter cake was dried in vacuo to give 2-(4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-yl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (50 mg, crude). The crude product was suspended in HCl / HO (4 N, 3 mL) and stirred at 20 °C for 12 h. The mixture was then diluted with DMSO (5 mL) and purified by prep-HPLC (column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: [water (HCl)-ACN]; B%: 15% to 45%, 10 min) to give (2-(4-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)phenyl)pyrimidin-5-yl)boronic acid (compound 24). LCMS: 367 (M+1). 1 H NMR: (400 MHz, DMSO-d6): δ= 9.16 (s, 2H), 8.54 (d, J = 8.5 Hz, 2H), 8.25 (d, J = 8.5 Hz, 2H), 3.55 (s, 2H), 2.92 (s, 4H).

[0809] compound 25 Synthesis of (3',5'-difluoro-4'-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1'-biphenyl]-4-yl)boronic acid (compound 25)

[0810] [ka]

[0811] Synthesis of 4'-bromo-3,5-difluoro-[1,1'-biphenyl]-4-carbonitrile

[0812] [ka]

[0813] Scheme 25A

[0814] [ka]

[0815] To a mixture of 2,6-difluoro-4-iodobenzonitrile (1) (5 g, 22.94 mmol, 1 eq) and 2-(4-bromophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1a) (6.49 g, 22.94 mmol, 1 eq) in dioxane (100 mL) and HO (10 mL) was added KCO (6.34 g, 45.87 mmol, 2 eq) and Pd(PPh)Cl (1.61 g, 2.29 mmol, 0.1 eq) under N at 20 °C. The mixture was degassed and purged with N three times, then the mixture was stirred at 110 °C under N for 16 h. The mixture was cooled to 20 °C and then diluted with water (100 mL). The mixture was then extracted with 300 mL of EtOAc (100 mL × 3). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue, which was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1). 4'-Bromo-3,5-difluoro-[1,1'-biphenyl]-4-carbonitrile (2) (1.8 g) was obtained. 1 H NMR: (400 MHz, CDCl3): δ= 7.57 (d, J = 8.6 Hz, 2H), 7.39 - 7.33 (m, 2H), 7.20 - 7.17 (m, 2H).

[0816] Synthesis of 4'-bromo-3,5-difluoro-[1,1'-biphenyl]-4-carboximidamide

[0817] [ka]

[0818] Scheme 25B

[0819] [ka]

[0820] To a solution of 4'-bromo-3,5-difluoro-[1,1'-biphenyl]-4-carbonitrile (2) (0.5 g, 1.70 mmol, 1 eq) in THF (5 mL) was added LiHMDS (1 M, 4.25 mL, 2.5 eq) dropwise at 0 °C. The mixture was stirred at 15 °C for 16 h. The reaction mixture was quenched by the dropwise addition of 4 N HCl / dioxane (10 mL). The mixture was concentrated under reduced pressure to give a residue. The residue was suspended in MeOH (10 mL) and stirred at 20 °C for 1 h. The mixture was then filtered, and the filtrate was concentrated under reduced pressure to give a residue. 4'-Bromo-3,5-difluoro-[1,1'-biphenyl]-4-carboximidamide (3) (528 mg, crude) was obtained, which was used in the next step without further purification.

[0821] Synthesis of 2-(4'-bromo-3,5-difluoro-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0822] [ka]

[0823] Scheme 25C

[0824] [ka]

[0825] To a mixture of 4'-bromo-3,5-difluoro-[1,1'-biphenyl]-4-carboximidamide-HCl (3) (528 mg, 1.52 mmol, 1 eq) in MeOH (10 mL) was added KCO (419.88 mg, 3.04 mmol, 2 eq) and methyl 4-oxotetrahydrothiopyran-3-carboxylate (396.97 mg, 2.28 mmol, 1.5 eq) at 25 °C under N. The mixture was then stirred at 25 °C for 16 h under N. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was triturated with water (10 mL) at 25 °C for 1 h and then filtered. The filter cake was dried in vacuo to give 2-(4'-bromo-3,5-difluoro-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (4) (0.9 g, crude), which was used directly in the next step.

[0826] Synthesis of 2-(3,5-difluoro-4'-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0827] [ka]

[0828] Scheme 25D

[0829] [ka]

[0830] To a mixture of 2-(4'-bromo-3,5-difluoro-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (4) (150 mg, 344.60 μmol, 1 eq) in dioxane (3 mL) was added B2Pin2 (131.26 mg, 516.90 μmol, 1.5 eq) and KOAc (101.46 mg, 1.03 mmol, 3 eq) at 25 °C. The mixture was degassed and purged with N three times, then Pd(dppf)Cl (28.14 mg, 34.46 μmol, 0.1 eq) was added under N at 25 °C. The mixture was stirred at 100 °C for 16 h under N atmosphere. After cooling to 20 °C, the reaction mixture was filtered and concentrated under reduced pressure to give a residue. The crude product was triturated with water (2 mL) and acetonitrile (2 mL) at 25 °C for 1 hour. The mixture was filtered, and the filter cake was dried in vacuo to give 2-(3,5-difluoro-4'-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (120 mg).

[0831] Scheme 25E

[0832] [ka]

[0833] To a stirred solution of HCl / HO (4 M, 2 mL), 2-(3,5-difluoro-4'-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (120 mg, 248.78 μmol, 1 eq) was added in portions at 20 °C under N. The mixture was then stirred at 50 °C under N for 2 h. The mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Waters Xbridge BEH C18 100 × 30 mm × 10 μm; mobile phase: [water (NHHCO)-ACN]; B%: 25% to 55%, 8 min). (3',5'-Difluoro-4'-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1'-biphenyl]-4-yl)boronic acid (compound 25) was obtained. LCMS: 401 (M+1). 1 H NMR: (400 MHz, DMSO-d6): δ= 13.30 - 12.89 (m, 1H), 8.19 (s, 2H), 7.98 - 7.86 (m, 2H), 7.84 - 7.77 (m, 2H), 7.73 - 7.65 (m, 2H), 3.55 (br s, 2H), 2.97 - 2.81 (m, 4H).

[0834] compound 26 Synthesis of (3'-hydroxy-4'-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1'-biphenyl]-4-yl)boronic acid (compound 26)

[0835] [ka]

[0836] Synthesis of 4'-bromo-3-methoxy-[1,1'-biphenyl]-4-carbonitrile

[0837] [ka]

[0838] Scheme 26A

[0839] [ka]

[0840] To a solution of 4-bromo-2-methoxybenzonitrile (1) (10.0 g, 47.2 mmol, 1.00 eq) and (4-bromophenyl)boronic acid (10.4 g, 51.9 mmol, 1.10 eq) in dioxane (50.0 mL) and HO (10.0 mL) was added KCO (13.0 g, 94.3 mmol, 2.00 eq) and Pd(dppf)Cl (3.45 g, 4.72 mmol, 0.10 eq) at 20 °C. The mixture was heated to 75 °C for 16 h. TLC (petroleum ether / ethyl acetate = 5 / 1, R f =0.53) indicated that 1 was completely consumed. The reaction mixture was diluted with EtOAc (50.0 mL) and HO (20.0 mL). The organic layer was separated from the mixture, and the aqueous phase was extracted with EtOAc (50.0 mL × 3). The combined organic layers were dried over NaSO, filtered, and concentrated under reduced pressure to give a residue, which was purified by column chromatography (SiO, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to give 4'-bromo-3-methoxy-[1,1'-biphenyl]-4-carbonitrile (2) (3.30 g, 8.02 mmol). The compound was used without further purification. 1 H NMR (400 MHz, DMSO-d6): δ= 7.86 (d, J = 8.1 Hz, 1H), 7.84 - 7.72 (m, 4H), 7.52 (d, J = 1.1 Hz, 1H), 7.45 (dd, J = 1.4, 8.0 Hz, 1H), 4.08 (s, 3H).

[0841] Synthesis of 4'-bromo-3-methoxy-[1,1'-biphenyl]-4-carboximidamide

[0842] [ka]

[0843] Scheme 26B

[0844] [ka]

[0845] To a solution of 4'-bromo-3-methoxy-[1,1'-biphenyl]-4-carbonitrile (2) (2.00 g, 6.94 mmol, 1.00 eq) in THF (20.0 mL) was added LiHMDS (1 M, 13.9 mL, 2.00 eq) dropwise at 0 °C, then warmed to 20 °C and stirred for 12 h. The reaction mixture was quenched with HCl / dioxane (13.0 mL) at 0 °C and then concentrated under reduced pressure to give a residue. The residue was triturated with MeOH (10 mL) and filtered. The filter cake was dried in vacuo to give a residue. 4'-Bromo-3-methoxy-[1,1'-biphenyl]-4-carboximidamide (3) (3.00 g, crude) was obtained. The crude product was used in the next step without further purification. LC-MS: 306 (M+1)

[0846] Synthesis of 2-(4'-bromo-3-methoxy-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0847] [ka]

[0848] Scheme 26C

[0849] [ka]

[0850] To a solution of 4'-bromo-3-methoxy-[1,1'-biphenyl]-4-carboximidamide (3) (3.00 g, 9.83 mmol, 1.00 eq) and methyl 4-oxotetrahydrothiopyran-3-carboxylate (3.43 g, 19.6 mmol, 2.00 eq) in MeOH (45.0 mL), KCO (5.43 g, 39.3 mmol, 4.00 eq) was added at 25 °C, then heated to 50 °C and stirred for 16 h. LC-MS showed that 3 was consumed. The reaction mixture was filtered, and the filter cake was dried under reduced pressure to obtain a residue. The residue was triturated with HO (30.0 mL) and stirred at 20 °C for 16 h. The mixture was then filtered, and the filter cake was dried under reduced pressure to give 2-(4'-bromo-3-methoxy-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (4) (1.5 g, crude), which was used in the next step without further purification. LC-MS: 430.2 (and bromo isomer) (M+1).

[0851] Synthesis of 2-(3-methoxy-4'-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0852] [ka]

[0853] Scheme 26D

[0854] [ka]

[0855] To a solution of 2-(4'-bromo-3-methoxy-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (4) (1.00 g, 2.33 mmol, 1.00 eq) and Pin2B2 (650 mg, 2.56 mmol, 1.10 eq) in dioxane (50.0 mL) was added KOAc (457 mg, 4.66 mmol, 2.00 eq) and Pd(dppf)Cl.CHCl (190 mg, 233 μmol, 0.10 eq) at 20 °C. The mixture was heated to 80 °C and stirred for 16 h. LC-MS showed that 4 was completely consumed, with one main peak of the desired mass being detected. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give 2-(3-methoxy-4'-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (1.60 g, crude). The crude product was used in the next step without further purification. LC-MS: 477.2 (M+1).

[0856] Synthesis of (3'-methoxy-4'-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1'-biphenyl]-4-yl)boronic acid

[0857] [ka]

[0858] Scheme 26E

[0859] [ka]

[0860] A mixture of 2-(3-methoxy-4'-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (1.60 g, 3.36 mmol, 1.00 eq) in HCl (8 M, 32.0 mL, 76.2 eq) was heated to 80 °C and stirred for 16 h. HCl (12 M, 8.00 mL, 28.6 eq) was added to the mixture at 20 °C, followed by heating to 80 °C for 4 h. LC-MS showed that 5 was completely consumed, with one main peak of the desired mass being detected. The reaction mixture was filtered, and the filter cake was dried under reduced pressure to give (3'-methoxy-4'-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1'-biphenyl]-4-yl)boronic acid (6) (710 mg, crude). The crude product was used in the next step without further purification. LC-MS: 395.2 (M+1).

[0861] Scheme 26F

[0862] [ka]

[0863] To a solution of (3'-methoxy-4'-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1'-biphenyl]-4-yl)boronic acid (6) (710 mg, 1.80 mmol, 1.00 eq) in DCM (21.6 mL) was added BBr3 (1 M in DCM, 9.00 mL, 5.00 eq) dropwise at -20 °C. The mixture was heated to 20 °C and stirred for 1 h. LC-MS showed that (6) was completely consumed, with one peak of the desired mass detected. The reaction mixture was quenched by adding ice water (20.0 mL) at 0 °C. The mixture was filtered, and the filter cake was dried under reduced pressure to give a residue, which was purified by prep-HPLC (column: Phenomenex Gemini NXC18 (75 × 30 mm × 3 um); mobile phase: [HO (0.05% NHHO + 10 mM NHHCO)-ACN]; gradient: 15% to 65% B over 8.0 min) to give (3′-hydroxy-4′-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1′-biphenyl]-4-yl)boronic acid (compound 26). LC-MS: 381.2 (M+1). 1 H NMR (400 MHz, DMSO-d6): δ= 8.27 (d, J = 8.1 Hz, 1H), 8.13 (s, 2H), 7.90 (d, J = 8.0 Hz, 2H), 7.71 (d, J = 8.0 Hz, 2H), 7.32 - 7.19 (m, 2H), 3.55 (s, 2H), 2.93 (s, 4H).

[0864] compound 27 Synthesis of (3-hydroxy-4'-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1'-biphenyl]-4-yl)boronic acid (compound 27)

[0865] [ka]

[0866] Synthesis of 4'-bromo-3'-methoxy-[1,1'-biphenyl]-4-carbonitrile

[0867] [ka]

[0868] Scheme 27A

[0869] [ka]

[0870] To a solution of 1-bromo-4-iodo-2-methoxybenzene (1) (8 g, 25.56 mmol, 1 eq) and (4-cyanophenyl)boronic acid (1a) (4.51 g, 30.7 mmol, 1.20 eq) in DMF (160 mL) was added CsCO (16.7 g, 51.1 mmol, 2.00 eq) and Pd(PPh) (1.48 g, 1.28 mmol, 0.05 eq) at 25 °C. The mixture was heated to 80 °C and stirred under N for 16 h. LC-MS showed that 1 was completely consumed. The reaction mixture was diluted with EtOAc and H2O. The organic layer was separated from the mixture, and the aqueous phase was extracted with EtOAc. The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to give 4'-bromo-3'-methoxy-[1,1'-biphenyl]-4-carbonitrile (2) (4.50 g, 15.6 mmol). 1 H NMR (400 MHz, DMSO-d6): δ= 8.00 (s, 4H), 7.74 (d, J = 8.2 Hz, 1H), 7.46 (d, J = 2.0 Hz, 1H), 7.31 (dd, J = 2.1, 8.2 Hz, 1H), 4.02 (s, 3H).

[0871] Synthesis of 4'-bromo-3'-methoxy-[1,1'-biphenyl]-4-carboximidamide

[0872] [ka]

[0873] Scheme 27B

[0874] [ka]

[0875] To a solution of 4'-bromo-3'-methoxy-[1,1'-biphenyl]-4-carbonitrile (2) (2.00 g, 6.94 mmol, 1.00 eq) in THF (20.0 mL) was added LiHMDS (1 M, 13.9 mL, 2.00 eq) dropwise at 0 °C. The mixture was warmed to 25 °C and stirred for 12 h. LC-MS showed that (2) was completely consumed. The reaction mixture was quenched by the dropwise addition of HCl / dioxane (4 M, 8.00 mL) at 0 °C and then concentrated under reduced pressure to give a residue. The residue was suspended in MeOH (20.0 mL), and the mixture was stirred at 25 °C for 1 h. The mixture was then filtered, and the filtrate was concentrated under reduced pressure to give 4'-bromo-3'-methoxy-[1,1'-biphenyl]-4-carboximidamide (3) (3.00 g, crude), which was used directly in the next step without further purification. LC-MS: 306.1 + bromo isomer (M+1).

[0876] Synthesis of 2-(4'-bromo-3'-methoxy-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0877] [ka]

[0878] Scheme 27C

[0879] [ka]

[0880] To a solution of 4'-bromo-3'-methoxy-[1,1'-biphenyl]-4-carboximidamide (3) (1.00 g, 3.28 mmol, 1.00 eq) and methyl 4-oxotetrahydrothiopyran-3-carboxylate (1.14 g, 6.55 mmol, 2 eq) in MeOH (15.0 mL) was added K2CO3 (1.81 g, 13.1 mmol, 4.00 eq) at 25 °C. The mixture was heated to 50 °C and stirred for 16 h. LC-MS showed that 3 was completely consumed. The reaction mixture was filtered, and the filter cake was dried under reduced pressure to obtain a residue. HO (10.0 mL) was added to the residue to form a slurry, which was stirred at 20 °C for 16 h. The mixture was then filtered, and the filter cake was dried under reduced pressure to give 2-(4'-bromo-3'-methoxy-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (4) (1.50 g, crude). LC-MS: 430.2 and bromo isomer (M+1). 1 H NMR (400 MHz, DMSO-d6): δ= 12.76 (br s, 1H), 8.26 - 8.20 (m, J = 8.4 Hz, 2H), 7.90 - 7.82 (m, J = 8.4 Hz, 2H), 7.68 (d, J = 8.3 Hz, 1H), 7.42 (d, J = 1.9 Hz, 1H), 7.27 (dd, J = 2.0, 8.3 Hz, 1H), 3.98 (s, 3H), 3.53 (s, 2H), 2.89 (s, 4H).

[0881] Synthesis of 2-(3'-methoxy-4'-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0882] [ka]

[0883] Scheme 27D

[0884] [ka]

[0885] To a solution of 2-(4'-bromo-3'-methoxy-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (4) (1.00 g, 2.33 mmol, 1.00 eq) and B2Pin2 (651 mg, 2.56 mmol, 1.10 eq) in dioxane (30.0 mL) was added KOAc (457 mg, 4.66 mmol, 2.00 eq) and Pd(dppf)Cl.DCM (190 mg, 233 μmol, 0.10 eq) at 25 °C. The mixture was heated to 80 °C and stirred under N for 16 h. LC-MS showed that 4 was completely consumed. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. 2-(3'-Methoxy-4'-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (1.00 g, crude) was obtained, which was used directly in the next step without further purification. LC-MS: 477.1 (M+1).

[0886] Synthesis of (3-methoxy-4'-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1'-biphenyl]-4-yl)boronic acid

[0887] [ka]

[0888] Scheme 27E

[0889] [ka]

[0890] 2-(3'-Methoxy-4'-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one (5) (1.00 g, 2.10 mmol, 1.00 eq) was added portionwise to a stirred solution of HCl / HO (4 M, 20.0 mL, 38.1 eq) at 20 °C. The mixture was stirred at 25 °C for 5 h. LC-MS showed that (5) was consumed. The reaction mixture was filtered, and the filter cake was dried under reduced pressure to give (3-methoxy-4'-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1'-biphenyl]-4-yl)boronic acid (6) (250 mg, crude), which was used directly in the next step. LC-MS: 395.2 (M+1).

[0891] Scheme 27F

[0892] [ka]

[0893] To a solution of (3-methoxy-4'-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1'-biphenyl]-4-yl)boronic acid (6) (250 mg, 634 μmol, 1.00 eq) in DCM (7.50 mL) was added BBr3 (1 M, 3.17 mL, 5.00 eq) dropwise at -20 °C. The mixture was stirred at 20 °C for 2 h. LC-MS showed that (6) was completely consumed. The reaction mixture was cooled to 0 °C, quenched with ice water (10.0 mL), and stirred at 0 °C for 10 min. The mixture was filtered, and the filter cake was dried under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex luna C18 100 × 40 mm × 5 μm; mobile phase: [HO (0.04% HCl)-ACN]; gradient: 10% to 50% B over 8.0 min) to give (3-hydroxy-4′-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1′-biphenyl]-4-yl)boronic acid (compound 27). LC-MS: 381.2 (M+1). 1 H NMR (400 MHz, DMSO-d6): δ= 8.24 - 8.07 (m, 2H), 7.94 - 7.84 (m, 1H), 7.83 - 7.74 (m, 2H), 7.37 - 7.04 (m, 2H), 3.55 (s, 2H), 2.92 (s, 4H).

[0894] compound 28 Synthesis of (2-hydroxy-4'-(4-oxo-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-2-yl)-[1,1'-biphenyl]-4-yl)boronic acid (compound 28)

[0895] [ka]

[0896] Synthesis of 4'-bromo-2'-methoxy-[1,1'-biphenyl]-4-carbonitrile

[0897] [ka]

[0898] Scheme 28A

[0899] [ka]

[0900] To a solution of 4-bromo-1-iodo-2-methoxybenzene (1) (8.00 g, 25.6 mmol, 1.00 eq) and (4-cyanophenyl)boronic acid (4.51 g, 30.7 mmol, 1.20 eq) in DMF (80.0 mL) was added CsCO (16.7 g, 51.1 mmol, 2.00 eq) and Pd(PPh) (1.48 g, 1.28 mmol, 0.05 eq) under N at 20 °C. The mixture was stirred at 80 °C under N for 14 h. TLC (petroleum ether / ethyl acetate = 8 / 1, R f = 0.54), indicating that 1 was completely consumed. The reaction mixture was diluted with EtOAc (50.0 mL) and HO (150 mL). The layers were separated, and the aqueous phase was extracted with EtOAc (50.0 mL × 3). The combined organic layers were dried over NaSO, filtered, and the filtrate was concentrated under reduced pressure to give a residue, which was purified by column chromatography (SiO, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to give 4'-bromo-2'-methoxy-[1,1'-biphenyl]-4-carbonitrile (2) (6.10 g, 21.2 mmol). 1 H NMR (400 MHz, DMSO-d6): δ= 7.93 (d, J = 8.4 Hz, 2H), 7.73 (d, J = 8.4 Hz, 2H), 7.42 (d, J = 1.4 Hz, 1H), 7.38 - 7.30 (m, 2H), 3.92 - 3.83 (m, 3H).

[0901] Synthesis of 4'-bromo-2'-methoxy-[1,1'-biphenyl]-4-carboximidamide

[0902] [ka]

[0903] Scheme 28B

[0904] [ka]

[0905] To a solution of 4'-bromo-2'-methoxy-[1,1'-biphenyl]-4-carbonitrile (2) (1.00 g, 3.47 mmol, 1.00 eq) in THF (10.0 mL) was added LiHMDS (1 M, 6.94 mL, 2.00 eq) dropwise at 0 °C. The mixture was stirred at 25 °C for 12 h. LC-MS showed that (2) was completely consumed. The reaction mixture was quenched by the dropwise addition of HCl / dioxane (6 mL) at 0 °C and then concentrated under reduced pressure to give a residue. This residue was suspended in MeOH (20 mL), and the mixture was stirred at 25 °C for 1 h. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give 4'-bromo-2'-methoxy-[1,1'-biphenyl]-4-carboximidamide (3) (2.00 g, crude), which was used in the next step without further purification. LC-MS: 306.2 + bromo isomer (M+1).

[0906] Synthesis of 2-(4'-bromo-2'-methoxy-[1,1'-biphenyl]-4-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one

[0907] [ka]

[0908] Scheme 28C

[0909] [ka]

[0910] To a solution of 4'-bromo-2'-methoxy-[1,1'-biphenyl]-4-carboximidamide (3) (1.00 g, 3.28 mmol, 1.00 eq) and methyl 4-oxotetrahydrothiopyran-3-carboxylate (1....

Claims

1. Formula (I): 【Chemical 1】 or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; (In the formula, R １ is hydrogen, deuterium, C １ -C ３ Alkyl, —OH, —O—C １ -C ３ Alkyl, —CH ２ OH, or -B(OH) ２ and R １ａ is hydrogen, deuterium, or C １ -C ３ is alkyl; R ２ teeth, (a) R ３ and optionally 1, 2 or 3 R ３ａ phenyl optionally substituted by a group; (b) phenyl substituted with a 5- or 6-membered heteroaryl, wherein the 5- or 6-membered heteroaryl is R ３ and optionally 1, 2 or 3 R ３ａ groups, and said phenyl may further optionally be substituted with one, two or three R ３ａ optionally substituted with a group), (c) optionally 1, 2, or 3 R ３ａ and may be substituted with a -phenyl-R ３ phenyl substituted with -phenyl-R ３ The phenyl in the group may optionally be substituted with one, two or three R ３ａ optionally substituted with a group; (d) R ３ and optionally 1, 2 or 3 R ３ａ 5- or 6-membered heteroaryl optionally substituted by a group; (e) optionally 1, 2, or 3 R ３ａ and may be substituted with a -phenyl-R ３ 5- or 6-membered heteroaryl substituted with 1, 2, or 3 R ３ａ optionally substituted with a group; (f) optionally 1, 2, or 3 R ３ａ and may be substituted with a -(5- or 6-membered heteroaryl)-R ３ 5- or 6-membered heteroaryl substituted with -(5- or 6-membered heteroaryl)-R ３ The 5- or 6-membered heteroaryl in ３ａ optionally substituted with a group; (g) R ３ and optionally one or two R ３ａ C optionally substituted with a group ３ -C ６ cycloalkyl; (h) C substituted with 5- or 6-membered heteroaryl ３ -C ６ cycloalkyl (wherein the 5- or 6-membered heteroaryl is R ３ and optionally 1, 2 or 3 R ３ａ groups, and the cycloalkyl may optionally be substituted with one or two R ３ａ optionally substituted with a group; (i) C substituted with phenyl ３ -C ６ cycloalkyl (wherein the phenyl is R ３ and optionally 1, 2 or 3 R ３ａ groups, and the cycloalkyl may optionally be substituted with one or two R ３ａ optionally substituted with a group; (j) 3- to 8-membered heterocycloalkyl substituted with phenyl or 5- or 6-membered heteroaryl, wherein the phenyl and the 5- to 6-membered heteroaryl are R ３ and optionally 1, 2 or 3 R ３ａ optionally substituted with a group; (k)-CH=CH-R ５ (Here, R ５ is phenyl or 5- or 6-membered heteroaryl, and said phenyl and said 5- or 6-membered heteroaryl are R ３ and optionally 1, 2 or 3 R ３ａ may be substituted with a group) and R ３ are independently -B(OH) ２ , cyano, halo, halo-C １ -C ６ Alkyl, -(C ０ -C ６ alkylene)-O-R ４ or a 5- to 10-membered heterocycle, wherein said 5- to 10-membered heterocycle is optionally substituted with cyano; or R ２ When is (a), R ３ and one R ３ａ When on adjacent carbons, they are combined with the carbon to which they are attached. 【Chemistry 2】 wherein the * indicates a carbon atom shared with the phenyl ring, and any remaining R ３ａ is as defined below, and each R ７ａ are independently hydrogen or C １ -C ６ alkyl) Forming; Each R ３ａ are independently cyano, halo, —OH, C １ -C ６ Alkyl, halo-C １ -C ６ Alkyl, and C １ -C ６ alkoxy; R ４ is hydroxy-C １ -C ６ Alkyl, C １ -C ６ Alkoxy-C １ -C ６ Alkyl, or C １ -C ６ Alkoxycarbonyl-NH-C １ -C ６ alkyl); (However, the compound is 2-(4-(trifluoromethyl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(4-(4-methoxyphenyl)piperazin-1-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(4-(3-methoxyphenyl)piperazin-1-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(4-(2-methoxyphenyl)piperazin-1-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(4-chlorophenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(5-(trifluoromethyl)pyridin-2-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(3-(trifluoromethyl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(3-(trifluoromethyl)phenyl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof; 2-(5-chlorothiophen-3-yl)-3,5,7,8-tetrahydro-4H-thiopyrano[4,3-d]pyrimidin-4-one, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof. (But not either).

2. R １ and R １ａ are independently selected from hydrogen and deuterium, or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof.

3. Ring A is 【Chemistry 3】 (In the formula, 【Chemistry 4】 indicates the bond to the remainder of the compound of formula (I) 3. The compound of claim 1 or 2, wherein:

4. R ２ But, R ３ and optionally 1, 2 or 3 R ３ａ 4. The compound of any one of claims 1 to 3, wherein R is phenyl optionally substituted by a group, or a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof.

5. R ３ is attached to the para position of the phenyl ring; or R ３ and one R ３ａ are located on adjacent carbon atoms, they form a ring (a-1) together with the carbon atoms to which they are attached, and The phenyl moiety may optionally be joined to the remaining R ３ａ optionally substituted with a group, 5. A compound of claim 4, or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof.

6. R ２ but, 【Chemistry 5】 or a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

7. R ２ but, 【Chemistry 6】 or a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

8. R ２ but, 【Chemistry 7】 or a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

9. R ２ is phenyl substituted with a 5- or 6-membered heteroaryl, wherein the 5- or 6-membered heteroaryl is R ３ and optionally 1, 2 or 3 R ３ａ groups, and said phenyl may further optionally be substituted with one, two or three R ３ａ 4. The compound of claim 1, wherein the compound is a methyl group, ...

10. R ２ is phenyl substituted at the para position with a 5- or 6-membered heteroaryl, wherein the 5- or 6-membered heteroaryl is R ３ and optionally 1, 2 or 3 R ３ａ groups, and said phenyl may further optionally be substituted with one, two or three R ３ａ and when said 5- or 6-membered heteroaryl is a 6-membered heteroaryl, R ３ is substituted at the para position of said 6-membered heteroaryl), or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof.

11. R ２ but, 【Chemistry 8】 11. The compound of any one of claims 1 to 3, 9, and 10, or a single stereoisomer or a mixture of stereoisomers thereof, and / or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

12. R ２ but, 【Chemistry 9】 or a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

13. R ２ optionally 1, 2 or 3 R ３ａ and may be substituted with a -phenyl-R ３ (wherein the -phenyl-R ３ The phenyl in ３ａ 4. The compound of claim 1, wherein the compound is a methyl group, ...

14. R ２ optionally 1, 2, or 3 R ３ａ group, and further at the para position, ３ (wherein the -phenyl-R ３ The phenyl may optionally be substituted with one, two or three R ３ａ may be substituted with a group, and R ３ is the -phenyl-R ３ 14. The compound of claim 13, or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof.

15. R ２ but, 【Chemistry 10】 or a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

16. R ２ but, 【Chemistry 11】 16. The compound of any one of claims 1 to 3 and 13 to 15, or a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

17. R ２ But, R ３ and optionally 1, 2 or 3 R ３ａ and optionally substituted with a 5- to 6-membered heteroaryl group (wherein R ３ is at the para position of said 6-membered heteroaryl), a compound of any one of claims 1 to 3, or a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof.

18. R ２ optionally 1, 2 or 3 R ３ａ and may be substituted with a -phenyl-R ３ wherein the phenyl is optionally substituted with 1, 2, or 3 R ３ａ optionally substituted with the -phenyl-R ３ is in the para position of said 6-membered heteroaryl; and optionally R ３ is in the para position of said phenyl), the compound of any one of claims 1 to 3, or a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof.

19. R ２ is a 5- or 6-membered heteroaryl, said 5- or 6-membered heteroaryl optionally containing 1, 2, or 3 R ３ａ and may be substituted with a -(5- or 6-membered heteroaryl)-R ３ wherein the -(5- or 6-membered heteroaryl)-R ３ The 5- or 6-membered heteroaryl in ３ａ group; optionally, the -(6-membered heteroaryl)-R ３ is in the para position of the first 6-membered heteroaryl; and optionally R ３ is in the para position of the 6-membered heteroaryl to which it is attached), a compound of any one of claims 1 to 3, or a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof.

20. R ２ but, 【Chemistry 12】 20. The compound of any one of claims 1 to 3 and 19, wherein:

21. R ２ But, R ３ and optionally one or two R ３ａ C optionally substituted with a group ３ -C ６ 4. The compound of any one of claims 1 to 3, wherein the compound is cycloalkyl, or a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof.

22. R ２ but, 【Chemistry 13】 (Optionally, where R ３ 22. The compound of any one of claims 1 to 3 and 21, or a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof.

23. R ２ is substituted with a 5- or 6-membered heteroaryl ３ -C ６ cycloalkyl (wherein the 5- or 6-membered heteroaryl is R ３ and optionally 1, 2 or 3 R ３ａ groups, and the cycloalkyl may optionally be substituted with one or two R ３ａ 4. The compound of claim 1, wherein the compound is a methyl group, ...

24. R ２ is substituted with phenyl ３ -C ６ cycloalkyl (wherein the phenyl is R ３ and optionally 1, 2 or 3 R ３ａ groups, and the cycloalkyl may optionally be substituted with one or two R ３ａ 4. The compound of claim 1, wherein the compound is a methyl group, ...

25. R ２ is a 3- to 8-membered heterocycloalkyl substituted with phenyl or a 5- or 6-membered heteroaryl, wherein said phenyl and said 5- to 6-membered heteroaryl are substituted with R ３ and optionally 1, 2 or 3 R ３ａ 4. The compound of claim 1, wherein the compound is a methyl group, ...

26. R ２ but, 【Chemistry 14】 26. The compound of any one of claims 1 to 3 and 25, selected from the group consisting of: or a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof.

27. R ２ is -CH=CH-R ５ (where R ５ is phenyl or 5- or 6-membered heteroaryl, and said phenyl and said 5- or 6-membered heteroaryl are R ３ and optionally 1, 2 or 3 R ３ａ 4. The compound of claim 1, wherein the compound is a methyl group, ...

28. R ２ but, 【Chemistry 15】 28. The compound of any one of claims 1 to 3 and 27, selected from the group consisting of: or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof.

29. R ３ But cyano, -B(OH) ２ , or -(C ０ -C ６ alkylene)-O-R ４ 29. The compound of any one of claims 1 to 28, wherein:

30. R ３ is cyano, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof.

31. R ３ But -B(OH) ２ or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof.

32. R ３ But -(C ０ -C ６ alkylene)-O-R ４ or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof.

33. R ３ But -(C １－６ alkylene)-O-R ４ or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof.

34. R ４ But hydroxy-C １ -C ６ 34. The compound of any one of claims 1 to 7, 9 to 11, 13 to 29, 32 and 33, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof.

35. R ４ But C １ -C ６ Alkoxy-C １ -C ６ 34. The compound of any one of claims 1 to 7, 9 to 11, 13 to 29, 32 and 33, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof.

36. R ４ But C １ -C ６ Alkoxycarbonyl-NH-C １ -C ６ 34. The compound of any one of claims 1 to 7, 9 to 11, 13 to 29, 32 and 33, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof.

37. R ２ is (b), (c), (e), or (f), and R ３ Halo, cyano, -B(OH) ２ , or -(C ０ -C ６ alkylene)-O-R ４ or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof.

38. 10. The compound of claim 1, selected from the group consisting of compounds 1-44 provided in Table 1, or a single stereoisomer or a mixture of stereoisomers thereof; a single tautomer or a mixture of tautomers thereof; and / or a pharmaceutically acceptable salt thereof.

39. 39. A pharmaceutical composition comprising a compound of any one of claims 1 to 38, or a single stereoisomer or a mixture of stereoisomers thereof, a single tautomer or a mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

40. The pharmaceutical carrier is a matrix component comprising a conjugate of graphene oxide (GO) and hyaluronic acid (HA) (GO-HA), where GO and HA are covalently bonded via a linker; Polyethylene glycol (PEG), wherein said PEG is optional; a thickening agent, wherein said thickening agent is optional; and water Including, 40. The pharmaceutical composition of claim 39, wherein optionally said compound comprises from about 0.001 wt% to about 5 wt% of the total composition.

41. 40. A method of inhibiting Wnt signaling pathway activity in a subject, comprising contacting the subject with an effective amount of a compound of any one of claims 1 to 38, or a stereoisomer or a single mixture of stereoisomers thereof, a tautomer or a single mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof.

42. 41. A method of treating a disease, disorder, or condition associated with Wnt signaling pathway activity, comprising administering to a mammal in need thereof a compound of any one of claims 1 to 38, or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof; or administering a pharmaceutical composition of claim 39 or 40.

43. 43. The method of claim 42, wherein the method is for stimulating tissue regeneration in a wound in a mammal in need thereof, and the wound is contacted with an effective amount of the compound (or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof) or the pharmaceutical composition.

44. 43. The method of claim 42, wherein the disease, disorder, or condition is selected from chronic wounds, acute wounds, alkali-burned corneal wounds, burns, lesions (including lesions caused by viruses selected from HPV and / or Poxviridae), inflammatory skin diseases (including acne, psoriasis, rosacea, and scleroderma), cartilage diseases (including osteoarthritis, rheumatoid arthritis, internal joint derangements, and degenerative cartilage diseases), bone diseases (including osteoporosis), organ fibrosis (including pulmonary fibrosis, cardiac fibrosis, liver fibrosis, kidney fibrosis), cancer (including melanoma, breast cancer, prostate cancer), denervated body parts in need of reinnervation, tissues in need of regeneration (including damaged elastic cartilage), bacterial growth in need of inhibition, fungal growth in need of inhibition, tissues in need of angiogenesis, osteoclast differentiation in need of inhibition, impaired osteoblast differentiation (where inhibition of osteoblast differentiation is necessary), and / or bone destruction associated with breast cancer.

45. 41. A method of inducing bacteriostasis associated with Wnt signaling pathway activity, comprising administering to a mammal in need thereof XAV939, or a tautomer and / or a pharmaceutically acceptable salt thereof, optionally in a pharmaceutically acceptable carrier; administering a compound of any one of claims 1 to 38, or a single stereoisomer or mixture of stereoisomers thereof, a single tautomer or mixture of tautomers thereof, and / or a pharmaceutically acceptable salt thereof; or administering a pharmaceutical composition of claim 39 or 40.

46. The following formula: 【Chemistry 16】 (In the formula, LG １ is a leaving group such as fluoro, chloro, bromo, iodo, triflate, mesylate, triazole, pyrazole, boronic acid, boronic ester, or allyltrifluoroborate; R １ is hydrogen, deuterium, C １ -C ３ Alkyl, —OH, —O—C １ -C ３ Alkyl, —CH ２ OH, or -B(OH) ２ and R １ａ is hydrogen, deuterium, or C １ -C ３ is alkyl; R ２０ is alkyl, preferably methyl or ethyl, or CD ３ and R ２ 'teeth, (b1) phenyl substituted with 5- or 6-membered heteroaryl, wherein the 5- or 6-membered heteroaryl is LG １ and optionally 1, 2 or 3 R ３ａ groups, and said phenyl may further optionally be substituted with one, two or three R ３ａ optionally substituted with a group), (c1) optionally 1, 2 or 3 R ３ａ and may be substituted with -phenyl-LG １ phenyl substituted with -phenyl-LG １ The phenyl in the group may optionally be substituted with one, two or three R ３ａ optionally substituted with a group; (e1) optionally 1, 2 or 3 R ３ａ and may be substituted with -phenyl-LG １ 5- or 6-membered heteroaryl substituted with 1, 2, or 3 R ３ａ optionally substituted with a group; (f1) optionally 1, 2 or 3 R ３ａ and may be substituted with a -(5- or 6-membered heteroaryl)-LG １ 5- or 6-membered heteroaryl substituted with -(5- or 6-membered heteroaryl)-LG １ The 5- or 6-membered heteroaryl in ３ａ optionally substituted with a group; (h1) NH ２ or OH, and optionally one or two R ３ａ C optionally substituted with a group ３ -C ６ cycloalkyl; or (i1) C substituted with phenyl ３ -C ６ cycloalkyl (wherein the phenyl is LG １ and said phenyl is further optionally substituted with 1, 2 or 3 R ３ａ groups, and the cycloalkyl may optionally be substituted with one or two R ３ａ may be substituted with a group) is) or a salt thereof, and / or a stereoisomer or mixture of stereoisomers; (However, the compound is: Methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate 【Chemistry 17】 or a salt thereof, and / or a stereoisomer or mixture of stereoisomers thereof).

46. a) Formula (A): 【Chemistry 18】 The compound of R ２ contacting with —C(O)H; or b) Formula (B): 【Chemistry 19】 The compound of R ２ '-C(NH)NH ２ contacting the compound with ２０ is Me or CD ３ or c) Formula (C): 【Chemistry 20】 The compound of R ２ contacting the compound with —H, １ is fluoro, chloro, bromo, iodo, triflate, mesylate, triazole, pyrazole, boronic acid, boronic ester, or allyltrifluoroborate; and optionally isolating the compound of formula (I); (In the formula, R ２ 'teeth, (b1) phenyl substituted with 5- or 6-membered heteroaryl, wherein the 5- or 6-membered heteroaryl is LG １ and optionally 1, 2 or 3 R ３ａ groups, and said phenyl may further optionally be substituted with one, two or three R ３ａ optionally substituted with a group), (c1) optionally 1, 2 or 3 R ３ａ and may be substituted with -phenyl-LG １ phenyl substituted with -phenyl-LG １ The phenyl in the group may optionally be substituted with one, two or three R ３ａ optionally substituted with a group; (e1) optionally 1, 2 or 3 R ３ａ and may be substituted with -phenyl-LG １ 5- or 6-membered heteroaryl substituted with 1, 2, or 3 R ３ａ optionally substituted with a group; (f1) optionally 1, 2 or 3 R ３ａ and may be substituted with a -(5- or 6-membered heteroaryl)-LG １ 5- or 6-membered heteroaryl substituted with -(5- or 6-membered heteroaryl)-LG １ The 5- or 6-membered heteroaryl in ３ａ optionally substituted with a group; (h1) NH ２ or OH, and optionally one or two R ３ａ C optionally substituted with a group ３ -C ６ cycloalkyl; or (i1) C substituted with phenyl ３ -C ６ cycloalkyl (wherein the phenyl is LG １ and said phenyl is further optionally substituted with 1, 2 or 3 R ３ａ groups, and the cycloalkyl may optionally be substituted with one or two R ３ａ may be substituted with a group) is) A process for preparing a compound of formula (I) according to any one of claims 1 to 37, comprising: (However, the compound Methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate 【Chemical Formula 21】 or a salt thereof and / or a stereoisomer or mixture of stereoisomers thereof).

47. 47. The method of claim 46, wherein said contacting is carried out under basic conditions.

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