Compositions and methods for regulating hair growth

Abstract

This application relates to compositions and methods for regulating hair growth. This disclosure relates to compounds capable of promoting hair growth or pharmaceutically acceptable salts thereof. This disclosure also relates to methods for promoting hair growth or treating disorders or conditions affecting hair growth, such as baldness or hair loss.

Description

[0001] This application is a divisional application of Chinese patent application No. 201880043696.2, entitled "Composition and Method for Regulating Hair Growth", which was filed on June 29, 2018, as PCT international application PCT / US2018 / 040385 and entered the Chinese national phase on December 27, 2019.

[0002] Related applications

[0003] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 527,775, filed June 30, 2017, and U.S. Provisional Patent Application No. 62 / 654,095, filed April 6, 2018. The contents of each of these applications are hereby incorporated by reference in their entirety. Technical Field

[0004] This application relates to methods for promoting hair growth or treating diseases or conditions that affect hair growth, such as baldness or hair loss. Background Technology

[0005] Hair follicle stem cells (HFSCs) undergo several successive quiescent phases (resting phases), interspersed with brief proliferative phases (telogen-anagen transitions) associated with the onset of the hair cycle. It is well known that the proliferation or activation of HFSCs is a prerequisite for the progression of the hair cycle. Despite advancements in treatment options, baldness and hair loss remain conditions that cannot be successfully treated in all individuals. Some existing treatments are inconvenient for users, while others require surgical intervention or other invasive procedures. Alternative therapies are needed. Summary of the Invention

[0006] In some respects, this disclosure provides compounds of formula I or II:

[0007]

[0008] Or its pharmaceutically acceptable salt.

[0009] in:

[0010] Each A is independently CH, CR 4 Or N;

[0011] Y is a carboxyl group, ester, amide, or...

[0012] Z represents CH and CR. 4 Or N.

[0013] R 2 It is CN or carboxyl group;

[0014] R 3It is H, aryl, aralkyl, or aralkylacyl, and optionally surrounded by one or more R groups. 5 Replace, where each R 5 Independently selected from alkyl, alkoxy, or halogen groups;

[0015] R 4 Each instance is independently an alkyl, carboxyl, halogen, hydroxyl, ester, or CN group;

[0016] R 6 Derived from H, alkyl, or cycloalkyl;

[0017] R 7 It is hydrogen, alkyl, halogen, hydroxyl, alkoxy, or acyloxy;

[0018] R 10 It is hydrogen or alkyl; and

[0019] n is 0-4.

[0020] In some embodiments, this disclosure provides compounds of formula III:

[0021]

[0022] Or its pharmaceutically acceptable salt.

[0023] in:

[0024] Y is a carboxyl group, ester, amide, or...

[0025] R 2 It is CN or carboxyl group;

[0026] R 3 It is H, aryl, aralkyl, or aralkylacyl, and optionally surrounded by one or more R groups. 5 Replace, where each R 5 Independently selected from alkyl, alkoxy, or halogen groups;

[0027] R 4 Each instance is independently an alkyl, carboxyl, halogen, hydroxyl, ester, or CN group;

[0028] R 6 Derived from H, alkyl or cycloalkyl; and

[0029] R 7 It is hydrogen, alkyl, halogen, hydroxyl, alkoxy, or acyloxy;

[0030] R 10 It is hydrogen or alkyl;

[0031] R 11 It is hydrogen or alkyl; and

[0032] n is 0-4.

[0033] In some embodiments, this disclosure provides compounds of formula V, VI, or VII:

[0034]

[0035] Or its pharmaceutically acceptable salt.

[0036] in:

[0037] Each A is independently CH, CR 4 Or N;

[0038] X is NR 6 Or O;

[0039] R 1 It is H or a lower alkyl group; or R 1 and R 6 Or R 1 and R 2 Together with the atoms that separate them, they complete a heterocycle;

[0040] R 2 It is CN or carboxyl group;

[0041] R 3 It is H, aryl, aralkyl, or aralkylacyl, and optionally surrounded by one or more R groups. 5 Replace, where each R 5 Independently selected from alkyl, alkoxy, and halogen groups;

[0042] R 4 Each instance is independently alkyl, carboxyl, halogen, hydroxyl, or CN;

[0043] R 6 Derived from H, alkyl or cycloalkyl; and

[0044] R 7 It is hydrogen, alkyl, halogen, hydroxyl, alkoxy, or acyloxy.

[0045] In some embodiments, this disclosure provides compounds of formula Va, VIa, or VIIa:

[0046]

[0047] and its pharmaceutically acceptable salt

[0048] in:

[0049] X is NR 6 Or O;

[0050] R 1 It is H or a lower alkyl group;

[0051] R 2 It is CN or carboxyl group; or R. 1 and R 2 Together with the atoms that separate them, they complete a heterocycle;

[0052] R 3 It is H, phenyl, or benzyl, and optionally surrounded by one or more R 5 Replace, where each R 5 Independently selected from alkyl, alkoxy, or halogen groups;

[0053] R 4 Each instance is independently selected from alkyl, carboxyl, halogen, hydroxyl, or CN groups; and

[0054] R 6 Selected from H, alkyl or cycloalkyl.

[0055] In some respects, this disclosure provides pharmaceutical compositions comprising compounds of this disclosure and pharmaceutically acceptable excipients.

[0056] In some respects, this disclosure provides a method for enhancing lactate production in cells, including contacting cells with compounds or compositions of this disclosure.

[0057] In some aspects, this disclosure provides a method for inhibiting mitochondrial pyruvate oxidation in cells, comprising contacting cells with mitochondrial pyruvate oxidation (MPO) inhibitors such as compounds disclosed herein. In some embodiments, the MPO inhibitor is a mitochondrial pyruvate carrier (MPC) inhibitor. In some embodiments, as described herein, inhibiting mitochondrial pyruvate oxidation in cells has the effect of enhancing lactate production in cells and / or enhancing LDH activity in cells and promoting hair growth.

[0058] In some aspects, this disclosure provides methods for enhancing lactate production in cells, including contacting cells with MPO inhibitors such as compounds disclosed herein. In some embodiments, the MPO inhibitor is a mitochondrial pyruvate carrier (MPC) inhibitor.

[0059] In some aspects, this disclosure provides methods for enhancing LDH activity in cells, including contacting cells with MPO inhibitors such as compounds disclosed herein. In some embodiments, the MPO inhibitor is a mitochondrial pyruvate carrier (MPC) inhibitor.

[0060] In some aspects, this disclosure provides methods for enhancing lactate dehydrogenase (LDH) activity in cells, including contacting cells with MPO inhibitors such as compounds disclosed herein. In some embodiments, the MPO inhibitor is a mitochondrial pyruvate carrier (MPC) inhibitor. In some aspects, this disclosure provides methods for promoting hair growth or treating hair growth disorders or conditions such as baldness or alopecia, including administering to a patient compounds or compositions as disclosed herein.

[0061] In some aspects, this disclosure provides methods for promoting hair growth or treating hair growth disorders or conditions such as baldness or alopecia, including administering to a patient an MPO inhibitor such as a compound of this disclosure (e.g., topical application, such as administration with a pharmaceutical composition formulated for topical application). In some embodiments, this disclosure provides methods for promoting hair growth or treating hair growth disorders or conditions such as baldness or alopecia, including administering to a patient an MPC inhibitor such as a compound of this disclosure (e.g., topical application, such as administration with a pharmaceutical composition formulated for topical application). In some embodiments, as described herein, inhibiting mitochondrial pyruvate oxidation or mitochondrial pyruvate carriers in cells has the effect of enhancing lactate production and / or enhancing LDH activity in cells and promoting hair growth.

[0062] In some respects, this disclosure provides compounds Or its pharmaceutically acceptable salt.

[0063] In some respects, this disclosure provides cosmetic methods for promoting hair growth, including administering compounds such as those disclosed herein to a subject.

[0064] In some respects, this disclosure provides cosmetic methods for treating diseases or conditions that affect hair growth, including administering compounds such as those disclosed to a subject.

[0065] In some respects, the ailment or condition described in this disclosure is baldness or hair loss.

[0066] In some respects, this disclosure provides for the use of compounds such as those disclosed in the manufacture of medicaments for promoting hair growth.

[0067] In some respects, this disclosure provides for the use of compounds such as those disclosed herein in the manufacture of medicaments for treating diseases or conditions affecting hair growth. Attached Figure Description

[0068] Figure 1A-1E The activity of lactate dehydrogenase was enriched in HFSC. Figure 1AIHC staining of Ldha expression throughout the hair cycle showed that Ldha protein was confined to the HFSC niche (raised area), indicated by brackets. IHC staining of Sox9 on serial sections delineated the HFSC population. Scale bar indicates 20 micrometers. Figure 1B Immunoblotting of FACS-separated HFSC populations (α6low / Cd34+ and α6hiCd34+) and total epidermis (Epi) revealed differential expression of Ldha in the stem cell niche. Sox9 is a biomarker for HFSC, and β-actin was used as a loading control. Figure 1C Colorimetric assays of Ldhase activity in the epidermis showed the highest activity in the raised areas (brackets) and the subcutaneous muscle layer (brackets). This activity was enriched in the raised areas at different stages of the hair cycle. Activity is indicated by purple; pink is nuclear counterstaining. It should also be noted that in stained mice, developing hair shafts showed the same intense melanin deposition as observed here. Hair shafts never showed any purple spots indicative of Ldh activity. Scale bar indicates 50 micrometers. Figure 1D Ldh activity in sorted cell populations, measured using a plate reader-based assay, also showed the highest Ldh activity in two separate HFSC populations (α6hi / Cd34 and α6low / Cd34) compared to epidermal cells (Epi) and fibroblasts (FBs). Each bar represents the mean signal for each cell type, where n = 9 mice were collected from 3 independent experiments. Results are shown as mean ± SEM. Paired t-tests were performed, with p < 0.05 for each cell type compared to epidermal cells. Figure 1E During the resting period (day 50), HFSCs and epidermal cells were isolated by FACS, and metabolites were extracted and analyzed by LC-MS. The heatmap shows the relative levels of glycolytic metabolites and TCA-circulating metabolites in cells isolated from different mice in independent experiments, with cells from three animals in each experiment. An asterisk indicates a significant difference in metabolite levels between epidermal cells and HFSCs. Paired t-tests were performed on e; * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001, ns indicates p > 0.05, n = 9 mice collected from 3 independent experiments.

[0069] Figure 2A-2C The validation of key reagents and assays is demonstrated. Figure 2A IHC, antibody specifically recognizes Ldha (and) Figure 1A (The same is used). Bottom image, IHC, antibody recognizes multiple isoforms of the Ldh protein. Scale bar indicates 20 micrometers. Figure 2B A classification strategy used to separate two cell populations from a protrusion. This special classification is used to separate, for example,... Figure 1B The protein sample is shown in the protein blot. Figure 2C Validation of the colorimetric assay for Ldh enzyme activity. The highest Ldh enzyme activity was observed in HFSC protrusions and muscle. Activity is indicated by purple staining; pink is nuclear solid red counterstaining. No detectable activity was observed in the absence of substrate lactate (purple staining). Right panel, further validation of the colorimetric assay for Ldh enzyme activity. Treatment of skin with HCl before adding the staining solution containing substrate lactate inhibited enzyme activity. No Ldh activity was detected (purple staining). Skin where enzyme activity was not inhibited by hydrochloric acid (HCl) showed the highest Ldh enzyme activity in HFSC protrusions and muscle. Scale bar indicates 50 micrometers.

[0070] Figures 3A-3E This indicates that Ldh activity increases during HFSC activation. Figure 3A GSEA analysis of RNA-seq transcriptome data from HFSC and total epidermis showed that glycolysis-related genes were enriched in HFSC (NES = 1.72). Figure 3B GSEA analysis of microarray transcriptome data from HFSC and total epidermis showed enrichment of glycolysis-related genes in HFSC (NES = 1.45). Results were generated from three mice in each case. Figure 3C RNA-seq data from HFSCs sorted during the resting or rest-growth phase transition showed that Ldha 21 The data represent the average values ​​of three independent animals at each time point. Figure 3D Ldh activity in sorted stem cell populations was measured using a plate reader-based assay. Results showed elevated Ldh activity due to stem cell activation during the telogen-ana phase transition. Each bar represents the mean signal for each condition, where n = 9 mice were collected from 3 independent experiments. Results are shown as mean ± SEM. Paired t-tests were performed, and p < 0.05 was observed. Figure 3E : A heatmap showing the relative levels of glycolytic metabolites and TCA-circulating metabolites extracted from HFSCs in a quiescent state (resting phase, day 50), an activated state (resting-growth phase, day 70), and a state restored to quiescence (growth phase, day 90). The data shown were generated from n = 3 animals at each time point in 3 independent experiments.

[0071] Figures 4A-4B This demonstrates the validation of hair cycle phase measurements. Figure 4ARNA-seq data were analyzed to verify that HFSCs in the telogen-growth phase transition were indeed in this transition. The telogen-growth phase transition is known to be driven by Shh (targeting Gli factor) and Wnt (targeting Lef1, Axin, and Ccnd1) signaling and is associated with increased proliferation (Ki67 and Pcna). Additionally, Sox4 has previously been identified as a regulator of the telogen-growth phase transition. n = 3 mice at each time point. Data are shown as mean ± SEM. Paired t-tests were performed, p < 0.05. Figure 4B Ki-67 staining marks dividing cells at various stages of the hair cycle. Parentheses indicate HFSC niches. Scale bar indicates 100 micrometers.

[0072] Figure 5A-5G The absence of Mpc1 showed that lactate production and HFSC activation were increased. Figure 5A Mpc1fl / fl animals exhibit pigment deposition and hair growth, consistent with entering the growth cycle at week 8.5, while Mpc1+ / + animals do not exhibit dorsal pigment deposition and hair growth at this early stage. The animals shown represent at least 12 animals of each genotype. Figure 5B HFSC protrusions in Mpc1+ / + and Mpc1fl / fl mice were isolated using FACS, followed by Western blotting, which showed successful deletion of Mpc1 protein in the stem cell niche. β-actin was used as a loading control. Figure 5C Ldh activity in the sorted HFSC population was measured using a plate reader, showing increased activity in Mpc1fl / flHFSC compared to Mpc1+ / +HFSC. Each bar represents the mean signal for each genotype, with n = 9 mice collected from 3 independent experiments. Results are shown as mean ± SEM. Paired t-tests were performed, and p < 0.05 was observed. Figure 5D Histological studies of WT and Mpc1-deficient skin showed that the absence of Mpc1 induced an anagen phase. Scale bar indicates 100 micrometers. Quantitative analysis of the right-hand phenotype shows the percentage of dorsal hair follicles in the resting phase, the transition from resting to anagen phase, and the percentage of anagen dorsal hair follicles in Mpc1+ / + and Mpc1fl / fl mice (n = 250 follicles, 3 mice from each genotype). Shown as mean ± SEM. Paired t-test performed, p < 0.05. Figure 5EImmunohistochemical staining for Ki-67 (a proliferation marker active only in HFSCs at the onset of a new hair cycle) showed its presence only in Mpc1fl / fl HFSCs at week 8.5, consistent with their accelerated entry into a new hair cycle. Phospo-S6, another marker active only in HFSCs at the onset of a new hair cycle, was present only in Mpc1fl / fl HFSCs. Staining for Sox9 showed its presence in HFSCs in the Mpc1-deficient niche. Images were taken at 60x magnification. Figure 5F Mpc1 deletion in mice carrying the Lgr5CreER allele showed strong induction of the hair cycle. Note the red boxes indicating the areas of new hair growth. Results represent at least 9 animals for each genotype. Figure 5G : Quantify pigment deposition in a specified genotype in three independent litters of pups (n = 5 mice per genotype).

[0073] Figures 6A-6D This demonstrates the impact of long-term Mpc1 deletion in HFSC. Figure 6A Six months after the onset of Mpc1 deletion in HFSC (K15CrePR; Mpc1fl / fl), mice lacking Mpc1 showed no detrimental effects, as measured by hair cycle (left), pathology (middle, H and E), or HFSC staining (right, Sox9). The scale bar indicates 100 micrometers in the middle image and 50 micrometers in the right image. Images represent at least 12 animals per genotype. Figure 6B To demonstrate that Mpc1 deletion specifically promotes HFSC proliferation, we used K15CrePR carrying the lox-stop-lox-Tomato allele; Ldha fl / fl Mice were used to observe K15+HFSC and proliferation in the presence and absence of Mpc1 deletion (left). Furthermore, we used ires-GFP from the Lgr5CreER allele to target Ki-67 and GFP staining and look for colocalization in the presence and absence of Mpc1 deletion (right). White brackets indicate raised areas. Scale bar indicates 20 micrometers. Figure 6C Mpc1 deletion in mice carrying the Lgr6CreER allele showed no premature induction of the hair cycle. Figure 6D Ldh activity was measured in sorted HFSCs from control or Lgr6CreER-mediated Mpc1 deletion mice, showing increased activity in Mpc1-deficient cells. n = 6 mice per genotype were collected from two independent experiments. Results are shown as mean ± SEM. Paired t-tests were performed, p < 0.05.

[0074] Figures 7A-7D The pharmacological inhibition of Mpc1 showed that it promoted HFSC activation. Figure 7AAnimals treated topically with UK-5099 (20 μM) showed pigmentation and hair growth after 8 days of treatment, indicating entry into the anagen phase. After 14 days of treatment, the anagen phase was achieved, indicated by complete hair coverage. Mice treated topically with the mediator control showed no pigmentation or hair growth even after 12 days of treatment. The right image shows dermatopathology indicating that the UK-5099 animals entered an accelerated anagen phase at week 8, characterized by downward follicular growth and subcutaneous thickening, while the mediator control animals showed neither and remained in the telogen phase. The images shown represent at least 14 mice from 7 independent experiments. The scale bar indicates 100 micrometers. Figure 7B : A graph showing the time to phenotype observation in mice treated with the medium and UK-5099. n = 6 mice per condition. Shown as mean ± SEM. Figure 7C Ldhase activity assays in the epidermis showed strong activity in HFSCs in both the mediator control and UK-5099-treated animals. Ldhase activity was also observed in the interfollicular epidermis of UK-5099-treated animals. Ldh activity was indicated by purple staining; pink was nuclear solid red counterstaining. Scale bar indicates 50 micrometers. Figure 7D 48-hour lactate metabolomics analysis was performed on HFSCs isolated from UK-5099-treated skin; each bar represents the mean signal for each condition, where n = 9 mice were collected from 3 independent experiments. Results are shown as mean ± SEM. Paired t-tests were performed, p < 0.05.

[0075] Figure 8 This demonstrates the effects of certain Mpc1 inhibitors described in this article on lactate production.

[0076] Figure 9 This demonstrates the effects of certain Mpc1 inhibitors described in this article on lactate production.

[0077] Figure 10 The EC50 calculations for UK5099 and JXL020 are shown.

[0078] Figure 11 The Mpc1 inhibitor of the present invention is shown to induce hair growth.

[0079] Figure 12 This demonstrates the effects of certain Mpc1 inhibitors described in this article on lactate production.

[0080] Figure 13 The effects of certain Mpc1 inhibitors described in this paper on total cell counts, normalized to DMSO treatment, are shown.

[0081] Figure 14The effects of certain Mpc1 inhibitors described in this paper on cellular lactate production, normalized to DMSO treatment, are shown.

[0082] Figure 15 The effects of certain Mpc1 inhibitors described in this paper on total cell counts, normalized to DMSO treatment, are shown.

[0083] Figure 16 The effects of certain Mpc1 inhibitors described in this paper on cellular lactate production, normalized to DMSO treatment, are shown.

[0084] Figure 17 The effects of certain Mpc1 inhibitors described in this paper on total cell counts, normalized to DMSO treatment, are shown.

[0085] Figure 18 The effects of certain Mpc1 inhibitors described in this paper on cellular lactate production, normalized to DMSO treatment, are shown.

[0086] Figure 19 The effects of certain Mpc1 inhibitors described in this paper on total cell counts, normalized to DMSO treatment, are shown.

[0087] Figure 20 This demonstrates the role of MPC in the oxidation of pyruvate to acetyl-CoA. Detailed Implementation

[0088] In some respects, this disclosure provides compounds of formula I or II:

[0089]

[0090] Or its pharmaceutically acceptable salt.

[0091] in:

[0092] Each A is independently CH, CR 4 Or N;

[0093] Y is a carboxyl group, ester, amide, or...

[0094] Z represents CH and CR. 4 Or N.

[0095] R 2 It is CN or carboxyl group;

[0096] R 3 It is H, aryl, aralkyl, or aralkylacyl, and optionally surrounded by one or more R groups. 5 Replace, where each R 5 Independently selected from alkyl, alkoxy, or halogen groups;

[0097] R 4 Each instance is independently an alkyl, carboxyl, halogen, hydroxyl, ester, or CN group;

[0098] R 6 Derived from H, alkyl, or cycloalkyl;

[0099] R 7 It is hydrogen, alkyl, halogen, hydroxyl, alkoxy, or acyloxy;

[0100] R 10 It is hydrogen or alkyl; and

[0101] n is 0-4.

[0102] In some embodiments, the compound is a compound of formula I. In some embodiments, the compound is a compound of formula II.

[0103] In some embodiments of Formula I or II, Z is CH or N.

[0104] In some embodiments, this disclosure provides compounds of formula III or IV:

[0105]

[0106] Or its pharmaceutically acceptable salt.

[0107] in,

[0108] Y is a carboxyl group, ester, amide, or...

[0109] R 2 It is CN or carboxyl group;

[0110] R 3 It is H, aryl, aralkyl, or aralkylacyl, and optionally surrounded by one or more R groups. 5 Replace, where each R 5 Independently selected from alkyl, alkoxy, or halogen groups;

[0111] R 4 Each instance is independently an alkyl, carboxyl, halogen, hydroxyl, ester, or CN group;

[0112] R 6 Derived from H, alkyl or cycloalkyl; and

[0113] R 7 It is hydrogen, alkyl, halogen, hydroxyl, alkoxy, or acyloxy;

[0114] R 10 It is hydrogen or alkyl;

[0115] R 11 It is hydrogen or alkyl; and

[0116] n is 0-4.

[0117] In some embodiments, the compound is a compound of formula III.

[0118] In some embodiments of Formulas I, II, and III, Y is where Y is In some implementations, R 10 It is H. In some implementations, R 10 It is an alkyl group (e.g., ethyl). In some embodiments, Y is an ester or amide group.

[0119] In some embodiments of Formula I, II, or III, R 11 It is an alkyl group (e.g., methyl).

[0120] In some embodiments, this disclosure provides compounds of formula V, VI, or VII:

[0121]

[0122] Or its pharmaceutically acceptable salt.

[0123] in:

[0124] Each A is independently CH, CR 4 Or N;

[0125] X is NR 6 Or O;

[0126] R 1 It is H or a lower alkyl group; or R 1 and R 6 Or R 1 and R 2 Together with the atoms that separate them, they complete a heterocycle;

[0127] R 2 It is CN or carboxyl group;

[0128] R 3 It is H, aryl, aralkyl, or aralkylacyl, and optionally surrounded by one or more R groups. 5 Replace, where each R 5 Independently selected from alkyl, alkoxy, and halogen groups;

[0129] R 4 Each instance is independently alkyl, carboxyl, halogen, hydroxyl, or CN;

[0130] R 6 Derived from H, alkyl, or cycloalkyl;

[0131] R 7It is hydrogen, alkyl, halogen, hydroxyl, alkoxy, or acyloxy; and

[0132] n is 0-4.

[0133] In some embodiments, the compound is a compound of formula V. In some embodiments, the compound is a compound of formula VI. In some embodiments, the compound is a compound of formula VII.

[0134] In some embodiments of formulas I, II, III, V, VI, or VII, at least one A is N, preferably, no more than two A's are N. In some preferred embodiments, exactly one A is N, preferably, A and NR are N. 3 They bind to the same carbon.

[0135] In some embodiments, this disclosure provides compounds of formula Va, VIa, or VIIa:

[0136]

[0137] and its pharmaceutically acceptable salt

[0138] in:

[0139] X is NR 6 Or O;

[0140] R 1 It is H or a lower alkyl group;

[0141] R 2 It is CN or carboxyl group; or R. 1 and R 2 Together with the atoms that separate them, they complete a heterocycle;

[0142] R 3 It is H, phenyl, or benzyl, and optionally surrounded by one or more R 5 Replace, where each R 5 Independently selected from alkyl, alkoxy, or halogen groups;

[0143] R 4 Each instance is independently selected from alkyl, carboxyl, halogen, hydroxyl, or CN groups;

[0144] R 6 Selected from H, alkyl, or cycloalkyl; and

[0145] n is 0-4.

[0146] In some embodiments, the compound is a compound of formula Va. In some embodiments, the compound is a compound of formula VIa. In some embodiments, the compound is a compound of formula VIIa.

[0147] In some embodiments of formulas V, VI, VII, Va, VIa, or VIIa, X is NH. In some embodiments, X is O.

[0148] In some embodiments of formulas V, VI, VII, Va, VIa, or VIIa, R 1 It is H. In some implementations, R 1 It is a lower alkyl group. In some embodiments, R 1 and R 6 Together with the atoms that separate them, they complete a heterocycle (e.g., morpholino).

[0149] In some embodiments of formulas V, VI, VII, Va, VIa, or VIIa, R 6 It is hydrogen.

[0150] In some embodiments of formulas V, VI, VII, Va, VIa, or VIIa, R 2 It is CN. In some implementations, R 2 It is a carboxyl group. In some embodiments, R 1 and R 2 Together with the atoms that separate them, a heterocyclic group selected from thiazoline-2,4-dione-5-ylidene or 2-iminothiazoline-4-one-5-ylidene is formed.

[0151] In certain embodiments of formulas I, II, III, V, VI, VII, Va, VIa, or VIIa, R 3 It is H. In some implementations, R 3 It is phenyl. In some embodiments, R 3 It is a phenyl group and is affected by one or more R groups. 5 Replacement. In some implementations, R 3 By an R 5 Replace, and where R 5 It is an alkoxy group. In some embodiments, R 3 It is an aralkyl group (e.g., benzyl or phenylethyl). In some embodiments, R 3 It is an aralkyl acyl group (e.g., phenylacetyl). In some embodiments, R 3 It is benzyl. In some embodiments, R 3 It is benzyl and is reacted with one or more R 5 Replacement. In some implementations, R 3 It is an aralkyl group (e.g., benzyl or phenethyl) and is bound by one or more R 5 Substitution (preferably on the benzene ring). In some embodiments, R 3 It is an aralkyl acyl group (e.g., phenylacetyl) and is bound by one or more R 5Substitution (preferably on the benzene ring). In some embodiments, R 3 By one or two R 5 Replace, where each R 5 Independently selected from fluoroalkyl or fluorine. In some embodiments, R 3 By two R 5 Replace, and each of R 5 It is trifluoromethyl.

[0152] In some implementations of formula Va, VIa, or VIIa, n is 0.

[0153] In some preferred embodiments, this disclosure provides compounds of formula Vb.

[0154]

[0155] In some embodiments, this disclosure provides compounds of formula Vc:

[0156]

[0157] In some implementations of formula Va, VIa, VIIa or Vb, n is 1.

[0158] In some preferred embodiments, this disclosure provides compounds of formula Vd.

[0159]

[0160] In some embodiments, this disclosure provides compounds of formula Ve:

[0161]

[0162] In some implementations of formula Vb, Vd, or Ve, R 4 Selected from halogenated or haloalkyl groups. In some preferred embodiments, R 4 It is a halogen group (e.g., chlorine or bromine). In other preferred embodiments, R 4 It is a carboxyl group or an ester.

[0163] In some embodiments of formula VI or VIa, n is 0. In some embodiments, n is 2, and R 4 Selected from halogenated or haloalkyl groups.

[0164] In certain embodiments of formulas I, II, III, V, VI, VII, Va, Vb, Vc, Vd, Ve, VIa, or VIIa, R 7 It is hydrogen, hydroxyl, halogen (e.g., chlorine), or acyloxy (e.g., acetoxy). In some embodiments, R 7It is a hydroxyl group, a halogen group (e.g., chlorine), or an acyl group (e.g., acetoxy).

[0165] In some embodiments of formulas I, II, III, V, VI, VII, Va, Vb, Vc, Vd, Ve, VIa, or VIIa, the compound is not JXL001.

[0166] In some respects, the compounds disclosed herein are the compounds listed in Table 1.

[0167] Table 1: Exemplary Compounds of the Invention

[0168]

[0169]

[0170]

[0171]

[0172]

[0173]

[0174]

[0175]

[0176] In some aspects, this disclosure provides pharmaceutical compositions comprising compounds of this disclosure and pharmaceutically acceptable excipients. In some embodiments, the pharmaceutical composition is formulated for topical application.

[0177] In some aspects, this disclosure provides methods for enhancing lactate production in cells, including contacting the cells with compounds or compositions of this disclosure. In some embodiments, the cells are hair follicle stem cells.

[0178] In some respects, this disclosure provides methods for promoting hair growth or treating hair growth disorders or conditions such as baldness or hair loss, including administering to a patient a compound or composition as disclosed herein.

[0179] In some aspects, this disclosure provides a method for enhancing lactate production in cells, including contacting cells with a mitochondrial pyruvate carrier (MPC) inhibitor. In some embodiments, the MPC is MPC1.

[0180] In some aspects, this disclosure provides methods for promoting hair growth or treating hair growth disorders or conditions such as baldness or alopecia, including administering a mitochondrial pyruvate carrier (MPC) inhibitor to a patient (e.g., topical application, such as with a pharmaceutical composition formulated for topical application). In some embodiments, the MPC is MPC1.

[0181] In some respects, this disclosure provides a method for inhibiting mitochondrial pyruvate oxidation in cells, including contacting cells with a mitochondrial oxidation (MPO) inhibitor.

[0182] In some respects, this disclosure provides methods for promoting hair growth or treating hair growth disorders or conditions such as baldness or hair loss, including administering a mitochondrial oxidation (MPO) inhibitor to a patient (e.g., topical application, such as with a pharmaceutical composition formulated for topical application).

[0183] In some embodiments, the MPC or MPO inhibitor is a compound disclosed herein.

[0184] discuss

[0185] Numerous studies have revealed that hair follicle stem cells (HFSCs) possess unique gene expression characteristics compared to other hair follicle cells or interfollicular epidermal cells. Many of these characteristics are regulated by transcription factors, which have later been shown to play important roles in HFSC homeostasis.

[0186] Lactate dehydrogenase is most commonly encoded by the Ldha and dh genes in mammals. Its protein product forms a homo- or hetero-tetramer to catalyze the reduction of pyruvate to lactate in a NADH-dependent manner and to lactate in an NAD-dependent manner. + Lactic acid is oxidized to pyruvate. Immunostaining revealed that Ldha was enriched in situ in static HFSCs (resting phase). Figure 1A Immunohistochemical staining (IHC) was performed using antibodies that recognize both Ldha and Ldhb, and the results showed that only Ldha appeared to be located in the HFSC niche. Figure 2A ).

[0187] IHC analysis also showed that Ldha expression was enriched in HFSC(Sox9+) during all three stages of the hair cycle. Figure 1A Consistently, immunoblotting of lysates from sorted cells showed that, relative to total epidermis ( Figure 1B Strong expression of Ldha in basal HFSC (α6HiCD34+) and basal (α6LoCD34+) HFSC populations (sorting strategy in) Figure 2B (Overview in Chinese).

[0188] To determine whether Ldha expression patterns are related to Ldh enzyme activity, a colorimetric enzyme assay was used to assess Ldh activity in situ. Ldh activity assays are typically performed on protein lysates or aliquots using a plate reader, and are well-suited for in situ measurement on frozen tissue sections. It is important to note that because both in situ and in vitro Ldh activity assays use excess substrate (lactic acid), the results reflect the potential activity of Ldh, rather than its steady-state activity.

[0189] Applying this assay to skin samples demonstrated that Ldh activity was significantly higher in HFSCs, consistent with the expression pattern of Ldha. Figure 1C Furthermore, Ldh activity is enriched in HFSC throughout the hair cycle. Figure 1C As a control, assays performed in the absence of enzyme substrate (lactic acid) or on acid-treated tissues produced zero activity. Figure 2C To further validate these results, we sorted the epidermal population, generated cell lysates on the sorted cells, and performed similar colorimetric-based enzyme assays on the sorted cell lysates, which also showed increased Ldh activity in HFSCs. Figure 1D To better characterize the metabolism of HFSCs, we performed metabolomics analysis on a sorted population from mouse skin using liquid chromatography-mass spectrometry (LC-MS) (Figure 1e). In three independent experiments (isolated from different mice on different days), several glycolytic metabolites (including glucose / fructose-6-phosphate, fructose-bisphosphate, dihydroxyacetone phosphate, 3-phosphoglycerate, and lactate) were generally higher in HFSCs relative to total epidermis. Conversely, most TCA circulating metabolites did not show consistent differences between epidermis and HFSCs. Figure 1E These results collectively indicate that, although all cells in the epidermis extensively utilize the TCA cycle to generate energy, Ldha expression, Ldh activity, and glycolytic metabolism are also increased in HFSCs.

[0190] Therefore, measuring metabolism throughout the hair cycle will capture any dynamic changes in the HFSC associated with activation or quiescence. Analysis of RNA-seq data from HFSCs isolated during the telogen or telogen-anagen transition demonstrated that Ldha is not only the major Ldh isoform expressed in HFSCs (Figure 3), but is also induced during the telogen-anagen transition. Figure 3A and 3B(NIHGEOGSE67404 and GSE51635). To confirm that cells analyzed by RNA-seq were indeed in the resting phase or transitioning from resting to growth phase, key biomarkers of this transition were evaluated, including the Shh and Wnt pathways (Gli1, 2, 3; Lef1, Axin1, Axin2, Ccnd1) and proliferation markers (Ki-67, Pcna, and Sox4). Figure 4A ).

[0191] In vitro Ldh activity assays of sorted HFSC lysates revealed that moderate induction of Ldh activity was associated with the transition from the resting phase to the growth phase. Figure 3D Hair cycle staging was verified using Ki-67 immunostaining to determine HFSC activation. Figure 4B Furthermore, measurements of steady-state metabolites extracted from sorted HFSCs showed that lactate in HFSCs increased as they transitioned from the resting phase to the rest-growth phase, and then decreased again during the growth phase as HFSCs returned to quiescence. Figure 3E ).

[0192] To determine whether lactate production induction affects HFSC activation or the hair cycle, we hybridized K15CrePR animals with those of fluxed mitochondrial pyruvate carrier 1 (Mpc1) (K15CrePR; Mpc1). fl / fl Mpc1 and Mpc2 form a heterodimer to form the mitochondrial pyruvate carrier MPC, a transport protein on the inner mitochondrial membrane required for pyruvate entry into the mitochondria. Loss of function of Mpc1 has been shown to drive lactate production by enhancing the conversion of pyruvate to lactate via Ldh. Furthermore, inhibition of MPC leads to a reduction in mitochondrial pyruvate oxidation (MPO) to acetyl-CoA. Figure 20 ).

[0193] In animals lacking Mpc1 in the HFSC, we observed a strong acceleration of the hair cycle on the abdomen and back, exhibiting all the typical features of the telogen-anagen transition. Figure 5A (n = 12 littermates). By day 70, mifepristone-treated K15CrePR; MPC1 fl / fl The animal was the only one to show signs of dorsal growth. Western blotting of sorted HFSCs confirmed the loss of Mpc1 protein. Figure 5B Importantly, purified HFSC lacking Mpc1 showed a strong induction of Ldh activity. Figure 5C Quantitative analysis of the dorsal hair cycle in three littermates revealed strong growth phase induction in the dorsal skin lacking Mpc1. Figure 5D(right), and histological examination showed that the growth period induction was outwardly normal, with typical subcutaneous dilatation ( Figure 5D Immunostaining revealed that Mpc1-null HFSCs induced multiple hair cycle activation markers such as Ki-67 and pS6, while Sox9 expression remained unaffected. Figure 5E Pathological examination and Sox9 staining showed that long-term loss of Mpc1 did not lead to abnormal hair follicles or depletion of HFSCs. Figure 6A Furthermore, the deletion of Mpc1 with Lgr5CreER exhibited a phenotype very similar to that with K15CrePR. Figure 5F and 5G This confirmed that the deletion of this protein in HFSCs led to its activation (n=12 pairs of littermates). Finally, immunofluorescence of Ires-GF and Ki-67 in the PLgr5CreER transgene and K15CrePR;Mpc1 fl / fl Lineage tracing in lsl-Tomato mice also demonstrated that HFSCs did indeed proliferate after Mpc1 deletion induced by tamoxifen or mifepristone. Figure 6B ).

[0194] UK-5099 (also referred to as JXL001 in this paper) is a recognized pharmacological inhibitor of mitochondrial pyruvate carriers, known to promote lactate production under various conditions. UK-5099 has the following structure:

[0195]

[0196] Topical treatment with UK-5099 in animals during the resting phase (day 50) resulted in a strong acceleration of the hair cycle and less proliferation of the interfollicular epithelium. Figure 7A Quantification of the hair cycle in at least six pairs of animals (mediate comparison UK-5099) showed a strong acceleration of the hair cycle within a short period of 6–9 days. Figure 7B Similar to Mpc1 gene deletion, 48-hour pharmacological blockade of mitochondrial pyruvate carriers by UK-5099 during the resting phase promoted increased Ldh activity in HFSCs and interfollicular epidermis, consistent with increased lactate production capacity. Figure 7C Finally, metabolomics analysis showed that topical application of UK-5099 increased the total lactate level in sorted HFSCs. Figure 7D ).

[0197] The synthesized compounds can locally promote an increase in lactic acid levels, thereby driving the hair cycle.

[0198] This compound is typically prepared by reacting the corresponding aldehyde (e.g., 1-phenylindole-3-carboxaldehyde for JXL001) with ethyl cyanoacetate in the presence of 40% aqueous L-proline solution to uniquely produce the E-isomer of ethyl 2-cyano-3-(1-phenylindole-3-yl)propionate, such as JXL004. Mild hydrolysis of the ester with lithium hydroxide yields the E-isomer of the acid, such as JXL001. All other compounds are prepared using specific aldehydes via a similar method. The two heterocyclic compounds, JXL023 and JXL024, are prepared by the condensation of 1-phenyl-indole-3-carboxaldehyde with thiazolidin-2,4-dione and 2-iminothiazolidin-4-one. The structures of all compounds were determined using conventional organic chemistry methods, particularly high-field proton, carbon, and fluorine NMR spectroscopy. 3 J C-H Coupling measurements revealed that these compounds all possess E-stereochemistry regarding the key carbon-carbon double bond.

[0199] To determine whether these compounds promote cellular lactate production, we treated cultured epithelial cells with the compounds and measured lactate levels in the culture medium using the Nova Biomedical BioProfile Basic Analyzer. Briefly, cultured epithelial cells were treated with DMSO, UK-5099 (also known as JXL001), or certain exemplary compounds disclosed herein for 24–30 hours, and then the lactate level in the culture medium was measured and normalized to cell number and experimental duration to obtain the cellular lactate production rate (nmol lactate, million cells, hours). Results are shown in… Figure 8 and 9 middle.

[0200] The rate of lactate production in the treated cells is shown in Figure 8 In the middle. As expected, since they are UK-5099 analogs, most of the new compounds showed an increase in lactic acid production. Figure 10 As shown, individual measurements were performed to calculate the EC values ​​of some compounds. 50 .

[0201] To determine the efficacy of these compounds on the hair cycle, mice were shaved on day 50 after birth and treated topically every other day with the compounds disclosed herein suspended in a shampoo solution for 3 weeks. Figure 11 As shown, in in vitro assays, all analogues that promote lactic acid production were also shown to stimulate hair growth over a 2-week period.

[0202] Pharmaceutical Composition

[0203] The compositions and methods of the present invention can be used to treat individuals in need. In some embodiments, the individual is a mammal, such as a human or a non-human mammal. When administered to an animal (such as a human), the composition or compound is preferably administered as a pharmaceutical composition comprising, for example, a compound of the present invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are known in the art and include, for example, aqueous solutions (such as water or physiologically buffered saline) or other solvents or media (such as glycols, glycerols, oils (such as olive oil) or injectable organic esters). In a preferred embodiment, when such a pharmaceutical composition is administered to a human, particularly via invasive routes of administration (i.e., bypassing routes of transport or diffusion across the epithelial barrier, such as injection or implantation), the aqueous solution is pyrogen-free or substantially pyrogen-free. Excipients may be selected, for example, to achieve delayed release of the agent or to selectively target one or more cells, tissues, or organs. The pharmaceutical composition may be in the form of dosage units such as tablets, capsules (including dispersible capsules and gelatin capsules), granules, hydrophilic colloids for reconstitution, powders, solutions, syrups, suppositories, injections, etc. The composition can also be present in transdermal delivery systems, such as skin patches. The composition can also be present in solutions suitable for topical application, such as lotions, creams, or ointments.

[0204] Pharmaceutically acceptable carriers may contain physiologically acceptable agents that, for example, stabilize, increase solubility, or enhance the absorption of compounds (such as those of the present invention). Such physiologically acceptable agents include, for example, carbohydrates (such as glucose, sucrose, or dextran), antioxidants (such as ascorbic acid or glutathione), chelating agents, low molecular weight proteins, or other stabilizers or excipients. The choice of pharmaceutically acceptable carrier (including physiologically acceptable agents) depends, for example, on the route of administration of the composition. The formulation or pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (formulation) may also be a liposome or other polymer matrix in which compounds of the present invention may be incorporated. For example, liposomes containing phospholipids or other lipids are non-toxic, physiologically acceptable, and metabolizable carriers that are relatively easy to prepare and administer.

[0205] The term “pharmaceutically acceptable” as used in this article refers to compounds, materials, compositions, and / or dosage forms that, within a reasonable medical judgment, are suitable for contact with tissues in humans and animals without excessive toxicity, irritation, allergic reactions, or other problems or complications, and that are commensurate with a reasonable benefit / risk ratio.

[0206] The phrase “pharmaceutically acceptable carrier” as used herein refers to pharmaceutically acceptable materials, compositions, or media, such as liquid or solid fillers, diluents, excipients, solvents, or encapsulating materials. Each carrier must be “acceptable” in the sense that it is compatible with other components of the formulation and does not harm the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth gum; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, and corn oil. (10) Soybean oil; (11) Diols, such as propylene glycol; (12) Polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (13) Esters, such as ethyl oleate and ethyl laurate; (14) Agar; (15) Buffers, such as magnesium hydroxide and aluminum hydroxide; (16) Alginate; (17) Atherless water; (18) Isotonic saline; (19) Ringer's solution; (20) Ethanol; (21) Phosphate buffer solution; and (22) Other non-toxic and compatible substances used in pharmaceutical preparations.

[0207] The pharmaceutical composition (formulation) may be administered to a subject via any of a number of routes of administration, including, for example, oral (e.g., drenches of aqueous or non-aqueous solutions or suspensions applied to the tongue, tablets, capsules (including dispersible capsules and gelatin capsules), granules, powders, pellets, pastes); absorption through the oral mucosa (e.g., sublingual); subcutaneous; transdermal (e.g., as a patch applied to the skin); and topical (e.g., as a cream, ointment, or spray applied to the skin). The compound may also be formulated for inhalation. In some embodiments, the compound may be dissolved or suspended only in sterile water. Details of suitable routes of administration and compositions suitable therefor can be found, for example, in U.S. Patent Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970, and 4,172,896, and the patents referenced therein.

[0208] The formulation can be conveniently provided in unit dosage forms and can be prepared by any method known in the pharmaceutical field. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the host being treated and the specific route of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form is generally the amount of the compound that produces the therapeutic effect. Typically, this amount ranges from about 1% to about 99% of the active ingredient, preferably from about 5% to about 70%, and most preferably from about 10% to about 30%, in percentage terms.

[0209] Methods for preparing these formulations or compositions include the step of associating an active compound (such as the compound of the present invention) with a carrier and optionally one or more auxiliary components. Generally, formulations are prepared by homogeneously and closely associating the compound of the present invention with a liquid carrier or a finely powdered solid carrier, or both, and then shaping the product as needed.

[0210] Formulations of the present invention suitable for oral administration may be in the form of capsules (including dispersible capsules and gelatin capsules), flat capsules, pills, tablets, lozenges (using a flavoring matrix, typically sucrose and gum arabic or tragacanth gum), hydrophilic colloids, powders, granules, or as solutions or suspensions in aqueous or non-aqueous liquids, or as oil-in-water or water-in-oil emulsions, or as elixirs or syrups, or as soft lozenges (using an inert matrix, such as gelatin and glycerin, or sucrose and gum arabic), and / or as oral washes, each containing a predetermined amount of the compound of the present invention as an active ingredient. The compositions or compounds may also be administered in the form of large pills, syrups, or pastes.

[0211] To prepare solid dosage forms (capsules (including dispersible capsules and gelatin capsules), tablets, pills, sugar tablets, powders, granules, etc.) for oral administration, the active ingredient is mixed with one or more pharmaceutically acceptable carriers (such as sodium citrate or dicalcium phosphate) and / or any of the following substances: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and / or silica; (2) binders, such as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and / or gum arabic; (3) humectants, such as... (3) Glycerin; (4) Disintegrants, such as agar, calcium carbonate, potato or cassava starch, alginate, certain silicates and sodium carbonate; (5) Solution retarders, such as paraffin; (6) Absorption enhancers, such as quaternary ammonium compounds; (7) Wetting agents, such as cetyl alcohol and glyceryl monostearate; (8) Adsorbents, such as kaolin and bentonite; (9) Lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate and mixtures thereof; (10) Complexing agents, such as modified and unmodified cyclodextrins; and (11) Colorants. In the case of capsules (including dispersible capsules and gelatin capsules), tablets and pills, the pharmaceutical composition may also contain a buffer. Similar types of solid compositions may also be used as fillers in soft-filled and hard-filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol.

[0212] Tablets can be prepared by compression or molding, optionally with one or more excipients. Compressed tablets can be prepared using binders (e.g., gelatin or hydroxypropyl methylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium glycolate starch or croscarmellose sodium), surfactants, or dispersants. Molded tablets can be prepared by molding a mixture of powdered compounds wetted with an inert liquid diluent in a suitable machine.

[0213] Tablets and other solid dosage forms of pharmaceutical compositions, such as sugar tablets, capsules (including dispersible capsules and gelatin capsules), pills, and granules, may optionally be scored or prepared with coatings and shells (e.g., enteric coatings and other coatings known in the field of pharmaceutical formulation). They may also be formulated to allow for the slow or controlled release of the active ingredient therein using, for example, hydroxypropyl methylcellulose (in varying proportions to provide a desired release profile), other polymer matrices, liposomes, and / or microspheres. They may be sterilized, for example, by filtration through a bacterial trap filter or by incorporation with a sterilizing agent in the form of a sterile solid composition, which may be dissolved in sterile water or some other sterile injectable medium prior to use. These compositions may also optionally contain a light-blocking agent and may also be compositions that optionally release the active ingredient in a delayed manner only or preferably in a portion of the gastrointestinal tract. Examples of encapsulation compositions that may be used include polymeric substances and waxes. Where appropriate, the active ingredient may also be in the form of microcapsules having one or more of the excipients described above.

[0214] Liquid dosage forms suitable for oral administration include pharmaceutically acceptable emulsions, hydrophilic colloids for reconstitution, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, liquid dosage forms may also contain inert diluents commonly used in the art, such as water or other solvents, cyclodextrins and their derivatives, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitol, as well as mixtures thereof.

[0215] In addition to inert diluents, oral compositions may also contain adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, coloring agents, aroma agents and preservatives.

[0216] In addition to active compounds, suspensions may also contain suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum hydroxide, bentonite, agar and tragacanth gum, and mixtures thereof.

[0217] Dosage forms for topical or transdermal application include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalers. Active compounds can be mixed under aseptic conditions with pharmaceutically acceptable carriers and any necessary preservatives, buffers, or propellants.

[0218] In addition to active compounds, ointments, pastes, creams and gels may also contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starches, tragacanth gum, cellulose derivatives, polyethylene glycol, silicones, bentonite, silicic acid, talc and zinc oxide, or mixtures thereof.

[0219] In addition to the active compound, powders and sprays may also contain excipients such as lactose, talc, silica, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures of these substances. Sprays may also contain commonly used propellants such as chlorofluorocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

[0220] Transdermal patches offer the added advantage of controlled delivery of the compounds of this invention into the body. Such dosage forms can be prepared by dissolving or dispensing the active compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of this flux can be controlled by providing a rate-controlled membrane or by dispersing the compound in a polymer matrix or gel.

[0221] As used herein, the phrases “parenteral administration” and “via parenteral administration” refer to modes of administration other than enteral and local administration, typically by injection, and including but not limited to intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, tracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration comprise one or more active compounds combined with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersants, suspensions, emulsions, or sterile powders, which can be reconstituted into sterile injectable solutions or dispersions just before use. These injectable solutions or dispersions may contain antioxidants, buffers, antibacterial agents, solutes that make the formulation isotonic with the blood of the intended recipient, or suspending agents or thickeners.

[0222] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. For example, appropriate flowability can be maintained by using coating materials (such as lecithin), by maintaining the desired particle size (in the case of dispersions), and by using surfactants.

[0223] These compositions may also contain adjuvants, such as preservatives, wetting agents, emulsifiers, and dispersants. By including various antibacterial and antifungal agents (e.g., parabens, chlorobutanol, phenolic sorbic acid, etc.), protection against microbial activity can be ensured. It is also desirable to include isotonic agents in the composition, such as sugars and sodium chloride. Additionally, by including agents that delay absorption, such as aluminum monostearate and gelatin, the absorption of injectable drug forms can be prolonged.

[0224] In some cases, to prolong the action of a drug, it is often desirable to slow the absorption of subcutaneously or intramuscularly injected drugs. This can be achieved by using liquid suspensions of crystalline or amorphous materials with poor water solubility. Therefore, the absorption rate of a drug depends on its dissolution rate, which in turn depends on the crystal size and crystal form. Alternatively, delayed absorption of parenteral drug forms can be achieved by dissolving or suspending the drug in an oil-based medium.

[0225] Injectable reservoir formulations are prepared by microencapsulating the subject compound in a biodegradable polymer such as polylactide-polyglycolic acid. The drug release rate can be controlled based on the drug-to-polymer ratio and the properties of the specific polymer used. Other examples of biodegradable polymers include poly(orthoester) and poly(anhydride). Reservoir-type injectable formulations are also prepared by encapsulating the drug in liposomes or microemulsions that are compatible with body tissues.

[0226] For use in the methods of the present invention, the active compound may be provided as is or in the form of a pharmaceutical composition containing, for example, 0.1% to 99.5% (more preferably 0.5% to 90%) of the active ingredient and a pharmaceutically acceptable carrier.

[0227] Introduction methods can also be provided via rechargeable or biodegradable devices. In recent years, various sustained-release polymer devices have been developed and tested in vivo for the controlled delivery of drugs, including protein biopharmaceuticals. A variety of biocompatible polymers, including hydrogels, both biodegradable and non-degradable, can be used to form implants for the sustained release of compounds at specific target sites.

[0228] The actual dose level of the active ingredient in a pharmaceutical composition can be varied to obtain an amount of active ingredient that can effectively achieve the desired therapeutic response for a specific patient, composition, and administration method without causing toxicity to the patient.

[0229] The selected dose level depends on a variety of factors, including the activity of the specific compound or combination of compounds or its esters, salts or amides used, the route of administration, the time of administration, the excretion rate of the specific compound used, the duration of treatment, other drugs, compounds and / or materials used in combination with the specific composition used, the age, sex, weight, condition, general health status and prior medical history of the patient being treated, and similar factors known in the medical field.

[0230] A physician or veterinarian with ordinary skills in the art can readily determine and prescribe the desired therapeutically effective amount of a pharmaceutical composition. For example, a physician or veterinarian may start with a dose of the pharmaceutical composition or compound below the required dose level to achieve the desired therapeutic effect and gradually increase the dose until the desired effect is achieved. The term "therapeutically effective amount" means the amount of compound sufficient to elicit the desired therapeutic effect. Generally, it should be understood that the effective amount of a compound will vary depending on the subject's weight, sex, age, and medical history. Other factors affecting the effective amount may include, but are not limited to, the severity of the patient's condition, the condition being treated, the stability of the compound, and, if necessary, another class of therapeutic agents administered with the compound of the present invention. A larger total dose may be delivered by multiple administrations of the agent. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996), Harrison's Principles of Internal Medicine, 13th edition, 1814-1882, incorporated herein by reference).

[0231] Generally speaking, the appropriate daily dose of the active compound used in the compositions and methods of the present invention will be the minimum amount of compound that effectively produces a therapeutic effect. This effective dose usually depends on the factors mentioned above.

[0232] If desired, the effective daily dose of the active compound can be administered as one, two, three, four, five, six, or more sub-dose administered at appropriate intervals throughout the day, optionally in unit dosage form. In some embodiments of the invention, the active compound may be administered two or three times daily. In a preferred embodiment, the active compound will be administered once daily.

[0233] Patients receiving this treatment are any animals in need, including primates, especially humans, as well as other mammals such as horses, cattle, pigs, sheep, cats, and dogs; poultry; and pets in general.

[0234] In some embodiments, the compounds of the present invention may be used alone or in combination with another type of therapeutic agent.

[0235] This disclosure includes the use of pharmaceutically acceptable salts of the compounds of the present invention in the compositions and methods of the present invention. In some embodiments, the intended salts of the present invention include, but are not limited to, alkyl, dialkyl, trialkyl, or tetraalkylammonium salts. In some embodiments, the intended salts of the present invention include, but are not limited to, L-arginine, benzylbenzylamine, benzathine penicillin, betaine, calcium hydroxide, choline, tannin, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucosamine, hydrabamine, 1H-imidazolium, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In some embodiments, the intended salts of the present invention include, but are not limited to, Na, Ca, K, Mg, Zn, or other metal salts. In some embodiments, the intended salts of the present invention include, but are not limited to, 1-hydroxy-2-naphtholic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetaminobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, L-ascorbic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, capric acid, caproic acid, caproic acid, octanoic acid, carbonic acid, cinnamic acid, citric acid, and cyclamic acid. (acid), dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactosidic acid, gentian acid, d-glucoheponic acid, d-gluconic acid, d-glucuronic acid, glutamic acid, glutamate, glutaric acid, glycerophosphate, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, L-malic acid, malonic acid, mandelic acid, methanesulfonic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pyruvic acid, phosphoric acid, propionic acid, L-pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, L-tartaric acid, thiocyanate, p-toluenesulfonic acid, trifluoroacetic acid, and undecanoic acid salt.

[0236] Pharmaceutically acceptable acid addition salts can also exist in various solvate forms, such as with water, methanol, ethanol, dimethylformamide, etc. Mixtures of such solvates can also be prepared. These solvates can originate from crystallization solvents, be inherent in the preparation or crystallization solvent, or be insoluble in such solvents.

[0237] Wetting agents, emulsifiers and lubricants (such as sodium lauryl sulfate and magnesium stearate), as well as colorants, release agents, coating agents, sweeteners, flavorings and aromas, preservatives and antioxidants may also be present in the composition.

[0238] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and (3) metal chelating agents, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0239] definition

[0240] Unless otherwise defined herein, the scientific and technical terms used in this application will have the meanings commonly understood by one of ordinary skill in the art. Generally, the nomenclature and techniques used in conjunction with those described herein in the fields of chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics, and protein and nucleic acid chemistry are those well-known and commonly used in the art.

[0241] Unless otherwise stated, the methods and techniques of this disclosure are generally performed according to conventional methods well known in the art and as described in the various general and more specific references cited and discussed throughout this specification. See, for example, “Principles of Neural Science”, McGraw-Hill Medical, New York, NY (2000); Motulsky, “Intuitive Biostatistics”, Oxford University Press, Inc. (1995); Lodish et al., “Molecular Cell Biology, 4th Edition”, WHFreeman & Co., New York (2000); Griffiths et al., “Introduction to Genetic Analysis, 7th Edition”, WHFreeman & Co., NY (1999); and Gilbert et al., “Developmental Biology, 6th Edition”, Sinauer Associates, Inc., Sunderland, MA (2000).

[0242] Unless otherwise stated herein, chemical terms used herein are used in accordance with their conventional usage in the art, as illustrated in “The McGraw-Hill Dictionary of Chemical Terms”, edited by Parker S, McGraw-Hill, San Francisco, CA (1985).

[0243] All the foregoing above, as well as any other publications, patents, and published patent applications mentioned in this application, are expressly incorporated herein by reference. In case of any conflict, this specification, including its specific definitions, shall prevail.

[0244] As used herein, the term "agent" refers to a compound (such as an organic or inorganic compound, a mixture of compounds), a biological macromolecule (such as nucleic acids, antibodies, including portions thereof, as well as humanized, chimeric, and human antibodies and monoclonal antibodies, proteins or portions thereof, such as peptides, lipids, carbohydrates), or an extract made from biological material such as bacteria, plants, fungi, or animal (especially mammalian) cells or tissues. Agents include, for example, agents with known structures and agents with unknown structures. The ability of such agents to inhibit or promote AR degradation makes them suitable as "therapeutic agents" in the methods and compositions disclosed herein.

[0245] The terms “patient,” “subject,” or “individual” are used interchangeably and refer to humans or non-human animals. These terms include mammals such as humans, primates, livestock (including cattle, pigs, etc.), companion animals (e.g., dogs, cats, etc.), and rodents (e.g., mice and rats).

[0246] “Treatment” of a disease or patient refers to taking measures to obtain a beneficial or desired outcome, including clinical outcomes. As used herein and as is well known in the art, “treatment” is a means of obtaining a beneficial or desired outcome (including clinical outcomes). Beneficial or desired clinical outcomes may include, but are not limited to, relief or improvement of one or more symptoms or disorders, reduction of disease severity, stabilization of disease status (i.e., no worsening), prevention of disease spread, delay or slowing of disease progression, improvement or mitigation of disease status, and remission (whether partial or complete), whether detectable or undetectable. “Treatment” may also mean prolonged survival compared to expected survival without treatment.

[0247] The term "prevention" is well known in the art and is well understood in the art when used in relation to conditions such as local recurrence (e.g., pain), diseases such as cancer, symptoms such as heart failure, or any other medical condition, and includes the administration of a composition that, relative to a subject not receiving the composition, reduces the frequency of symptoms of the subject's medical condition or delays its onset. Thus, cancer prevention includes, for example, reducing, in statistically and / or clinically significant amounts, the number of detectable cancerous growths in a patient population receiving prophylactic treatment relative to an untreated control population, and / or delaying the occurrence of detectable cancerous growths in the treated population relative to the untreated control population.

[0248] The administration of a substance, compound, or agent to a subject may be carried out using one of a variety of methods known to those skilled in the art. For example, the compound or agent may be administered intravenously, intra-arterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, via the eyes, sublingually, orally (by ingestion), nasally (by inhalation), intraspinally, intracerebrally, and percutaneously (by absorption, e.g., via a skin tube). The compound or agent may also be suitably introduced via a rechargeable or biodegradable polymer device or other means such as patches and pumps, or by a formulation that provides a prolonged, slow, or controlled release of the compound or agent. Administration may also be performed, for example, once, multiple times, and / or over one or more extended time periods.

[0249] The appropriate method of administering a substance, compound, or agent to a subject will also depend on factors such as the subject's age and / or physical condition, and the chemical and biological properties of the compound or agent (e.g., solubility, digestibility, bioavailability, stability, and toxicity). In some embodiments, the compound or agent is administered orally, for example, by ingestion. In some embodiments, the orally administered compound or agent is in a prolonged-release or slow-release formulation, or is administered using a device for such slow or prolonged release.

[0250] As used herein, the phrase “combined administration” refers to the administration of any form of two or more different therapeutic agents such that the second agent is administered while the previously administered agent is still effective in the body (e.g., both agents are effective in the patient simultaneously, which may include a synergistic effect between the two agents). For example, different therapeutic compounds may be administered simultaneously or sequentially in the same formulation or as separate formulations. Thus, individuals receiving such treatment may benefit from the combined effect of different therapeutic agents.

[0251] The "therapeutic effective amount" or "therapeutic dose" of a drug or agent refers to the amount of drug or agent that will have the expected therapeutic effect when administered to a subject. A complete therapeutic effect may not necessarily occur with a single dose, but may only occur after a series of doses. Therefore, a therapeutic effective amount can be administered in one or multiple doses. The precise effective amount required by the subject will depend on factors such as the subject's size, health, and age, as well as the nature and extent of the condition being treated (such as cancer or MDS). Skilled technicians can easily determine the effective amount under given conditions through routine experiments.

[0252] The term "acyl" is well known in the art and refers to a group represented by the general formula hydrocarbon C(O)-, preferably alkyl C(O)-.

[0253] The term "acylamino" is well known in the art and refers to an amino group that has been substituted with an acyl group, and can be represented, for example, by the alkyl group C(O)NH-.

[0254] The term "acyloxy group" is well known in the art and refers to a group represented by the general formula hydrocarbon C(O)O-, preferably alkyl C(O)O-.

[0255] The term "alkoxy group" refers to an alkyl group with an oxygen atom attached to it. Representative alkoxy groups include methoxy, ethoxy, propoxy, and tert-butoxy.

[0256] The term "alkoxyalkyl" refers to an alkyl group that has been substituted with an alkoxy group, and can be represented by the general formula alkyl-O-alkyl.

[0257] The term "alkyl" refers to a saturated aliphatic group, including straight-chain alkyl, branched-chain alkyl, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl, and cycloalkyl-substituted alkyl. In a preferred embodiment, the straight-chain or branched alkyl group has 30 or fewer carbon atoms in its main chain (e.g., a straight chain of C16). 1-30 The branch is C 3-30 ), and more preferably 20 or fewer.

[0258] Furthermore, the term "alkyl" as used throughout the specification, examples, and claims is intended to include both unsubstituted and substituted alkyl groups, the latter referring to an alkyl moiety having a substituent of substituted hydrogen on one or more carbon atoms of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl.

[0259] Term "C" x-y "or "C x -C y"When used in conjunction with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, it is intended to include groups containing x to y carbons in the chain. C0 alkyl indicates a hydrogen atom at the terminal position, and if internal, it indicates a bond. For example, C..." 1-6 Alkyl groups contain 1 to 6 carbon atoms in the chain.

[0260] As used herein, the term "alkylamino" refers to an amino group that is substituted with at least one alkyl group.

[0261] As used herein, the term "alkylthio" refers to a thiol group substituted with an alkyl group and can be represented by the general formula alkylS-.

[0262] As used herein, the term "amide" refers to a group.

[0263]

[0264] Where R 9 and R 10 Each can independently represent a hydrogen or hydrocarbon group, or R 9 and R 10 Together with the N atoms they are attached to, they complete heterocycles with 4 to 8 atoms in the ring structure.

[0265] The terms "amine" and "amino" are well known in the art and refer to unsubstituted and substituted amines and their salts, such as those represented by the following portions.

[0266]

[0267] Where R 9 R 10 and R 10 Each can independently represent a hydrogen or hydrocarbon group, or R 9 and R 10 Together with the N atoms they are attached to, they complete heterocycles with 4 to 8 atoms in the ring structure.

[0268] As used herein, the term "aminoalkyl" refers to an alkyl group that has been substituted with an amino group.

[0269] As used herein, the term "aralkyl" refers to an alkyl group that has been substituted with an aryl group.

[0270] As used herein, the term "aryl" includes a substituted or unsubstituted monocyclic aromatic group, wherein each atom of the ring is a carbon. Preferably, the ring is a 5- to 7-membered ring, more preferably a 6-membered ring. The term "aryl" also includes a polycyclic system having two or more rings, wherein two or more carbons are shared by two adjacent rings, and wherein at least one ring is aromatic; for example, the other rings may be cycloalkyl, cycloalkenyl, cycloynyl, aryl, heteroaryl, and / or heterocyclic. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, etc.

[0271] The term "carbamate" is well known in the art and refers to a group...

[0272]

[0273] Where R 9 and R 10 Independently represents either hydrogen or hydrocarbon groups.

[0274] As used herein, the term "carbocyclic alkyl" refers to an alkyl group that has been substituted with a carbocyclic group.

[0275] As used herein, the terms "carbocycle," "carbocyclic group," and "carbocyclic ring" refer to a non-aromatic saturated or unsaturated ring in which each atom of the ring is carbon. Preferably, the carbocycle contains 3 to 10 atoms, more preferably 5 to 7 atoms.

[0276] As used herein, the term "carbocyclic alkyl" refers to an alkyl group that has been substituted with a carbocyclic group.

[0277] The term "carbonate" is well known in the art and refers to the group -OCO2-.

[0278] As used herein, the term "carboxyl" refers to a group represented by the formula -CO2H.

[0279] As used herein, the term "ester" refers to the group -C(O)OR 9 , where R 9 It indicates a hydrocarbon group.

[0280] As used herein, the term "ether" refers to a hydrocarbon group that is attached to another hydrocarbon group via oxygen. Therefore, the ether substituent of a hydrocarbon group can be alkyl-O-. Ethers can be symmetrical or asymmetrical. Examples of ethers include, but are not limited to, heterocyclic-O-heterocycles and aryl-O-heterocycles. Ethers include "alkoxyalkyl," which can be represented by the general formula alkyl-O-alkyl.

[0281] As used herein, the terms “halogen” and “halogen” refer to halogens and include chlorine, fluorine, bromine, and iodine.

[0282] As used herein, the terms “heteroalkyl” and “heteroarylalkyl” refer to alkyl groups that have been substituted with heteroaryl groups.

[0283] The term "heteroaryl" includes substituted or unsubstituted aromatic monocyclic structures, preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings, whose ring structure includes at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The term "heteroaryl" also includes polycyclic systems having two or more rings, wherein two or more carbons are shared by two adjacent rings, wherein at least one ring is heteroaromatic; for example, the other rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclic groups. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine.

[0284] As used herein, the term "heteroatom" refers to an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0285] As used herein, the term "heterocyclic alkyl" refers to an alkyl group that has been substituted with a heterocyclic group.

[0286] The terms "heterocyclic group," "heterocyclic," and "heterocyclic" refer to substituted or unsubstituted non-aromatic ring structures, preferably 3 to 10-membered rings, more preferably 3 to 7-membered rings, whose ring structure includes at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heterocyclic group" and "heterocyclic" also include polycyclic systems having two or more rings, wherein two or more carbons are shared by two adjacent rings, and wherein at least one ring is heterocyclic; for example, the other rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclic groups. Heterocyclic groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactone, lactam, etc.

[0287] As used herein, the term "hydrocarbon group" refers to a group bonded by carbon atoms without =O or =S substituents and typically has at least one carbon-hydrogen bond and a predominant carbon backbone, but may optionally include heteroatoms. Therefore, for the purposes of this application, groups such as methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered hydrocarbon groups, but substituents such as acetyl (which has a =O substituent on the linked carbon) and ethoxy (which is linked by oxygen rather than carbon) are not. Hydrocarbon groups include, but are not limited to, aryl, heteroaryl, carbocyclic, heterocyclic, alkyl, alkenyl, ynyl, and combinations thereof.

[0288] As used herein, the term "hydroxyalkyl" refers to an alkyl group that has been substituted with a hydroxyl group.

[0289] When used in conjunction with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, the term "lower" means a group comprising ten or fewer atoms, preferably six or fewer, of which the substituents. For example, "lower alkyl" refers to an alkyl group containing ten or fewer, preferably six or fewer carbon atoms. In some embodiments, the acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents as defined herein are lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, whether they appear alone or in combination with other substituents, such as hydroxyalkyl and aralkyl in the description (in which case, for example, atoms within the aryl group are not counted when calculating the carbon atoms in the alkyl substituent).

[0290] The terms "polycyclic," "polycyclic," and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclic) in which two or more atoms are shared by two adjacent rings; for example, these rings are "fused rings." Each ring of a polycyclic compound may be substituted or unsubstituted. In some embodiments, each ring of the polycyclic compound contains 3 to 10 atoms, preferably 5 to 7 atoms.

[0291] The term “sulfate” is well known in the art and refers to the group -OSO3H or its pharmaceutically acceptable salt.

[0292] The term "sulfonamide" is well known in the art and refers to a group represented by the following general formula.

[0293]

[0294] Where R 9 and R 10 Independently represents either hydrogen or hydrocarbon groups.

[0295] The term "sulfoxide" is well known in the art and refers to the group -S(O)-.

[0296] The term "sulfonate" is well known in the art and refers to the group SO3H or its pharmaceutically acceptable salt.

[0297] The term "sulfone" is well known in the art and refers to the group –S(O)2-.

[0298] The term "substituted" refers to a portion having a substituent that replaces hydrogen on one or more carbons of the main chain. It should be understood that "substituted" or "replaced by" includes the implicit condition that such substitution meets the permissible valence of the substituted atom and the substituent, and that the substitution produces a stable compound, such as a compound that is not spontaneously transformed by rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is contemplated to include all permissible substituents in organic compounds. In a broad sense, permissible substituents include acyclic and cyclic, branched or unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents in organic compounds. For suitable organic compounds, permissible substituents can be one or more identical or different substituents. For the purposes of this invention, heteroatoms (such as nitrogen) may have hydrogen substituents and / or any permissible substituent in the organic compounds described herein that satisfies the valence of the heteroatom. Substituents may include any substituents described herein, such as halogens, hydroxyl groups, carbonyl groups (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl groups (such as thioesters, thioacetic acids, or thiocarbamates), alkoxy groups, phosphoryl groups, phosphate groups, phosphonate groups, hypophosphonate groups, amino groups, amide groups, amidine groups, imine groups, cyano groups, nitro groups, azide groups, mercapto groups, alkylthio groups, sulfate groups, sulfonate groups, aminosulfonyl groups, sulfonamide groups, sulfonyl groups, heterocyclic groups, aralkyl groups, or aromatic or heteroaromatic moieties. Those skilled in the art will understand that, where appropriate, the substituted portion of the hydrocarbon chain may itself be substituted.

[0299] As used herein, the term "thioalkyl" refers to an alkyl group that has been substituted with a thiol group.

[0300] As used herein, the term "thioester" refers to the group -C(O)SR. 9 Or –SC(O)R 9

[0301] Where R 9 It indicates a hydrocarbon group.

[0302] As used in this article, the term "thioether" is equivalent to ether, in which oxygen is replaced by sulfur.

[0303] The term "urea" is well known in the art and can be represented by the following general formula.

[0304]

[0305] Where R 9 and R 10 Independently represents either hydrogen or hydrocarbon groups.

[0306] As used herein, the term “regulation” includes both inhibition / suppression of function or activity (such as cell proliferation) and enhancement of function or activity.

[0307] The phrase "pharmaceutically acceptable" is well known in the art. In some embodiments, the term includes, within the bounds of reasonable medical judgment, compositions, excipients, adjuvants, polymers, and other materials and / or dosage forms that come into contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, and in proportion to a reasonable benefit / risk ratio.

[0308] "Pharmaceutically acceptable salt" or "salt" as used herein refers to an acid addition salt or base addition salt that is suitable for or compatible with the treatment of a patient.

[0309] As used herein, the term "pharmaceutically acceptable acid addition salt" means any non-toxic organic or inorganic salt of any base compound represented by Formula I or II. Exemplary inorganic acids that form suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid, as well as metal salts such as sodium orthophosphate and potassium hydrogen sulfate. Exemplary organic acids that form suitable salts include monocarboxylic acids, dicarboxylic acids, and tricarboxylic acids, such as glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, benzoic acid, phenylacetic acid, cinnamic acid, and salicylic acid, as well as sulfonic acids such as p-toluenesulfonic acid and methanesulfonic acid. Monocarboxylic acid salts or dicarboxylic acid salts can be formed, and such salts can exist in hydrated, solvated, or substantially anhydrous forms. Generally, acid addition salts of compounds of Formula I or II are more soluble in water and a variety of hydrophilic organic solvents and typically exhibit higher melting points compared to their free base forms. Suitable salt selection is known to those skilled in the art. Other non-pharmaceutical acceptable salts, such as oxalates, can be used, for example, for the separation of compounds of formula I or II for laboratory use, or subsequently converted into pharmaceutically acceptable acid addition salts.

[0310] As used herein, the term "pharmaceutically acceptable base addition salt" means any non-toxic organic or inorganic base addition salt of any acid compound or any intermediate thereof represented by Formula I or II. Exemplary inorganic bases that form suitable salts include lithium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, or barium hydroxide. Exemplary organic bases that form suitable salts include aliphatic, alicyclic, or aromatic organic amines, such as methylamine, trimethylamine, and methylpyridine or ammonia. The selection of suitable salts is known to those skilled in the art.

[0311] Many compounds that can be used in the methods and compositions of this disclosure have at least one stereocenter in their structure. This stereocenter may be present in an R or S configuration, and the R and S symbols are used in accordance with the rules described in Pure Appl. Chem. (1976), 45, 11-30. This disclosure covers all stereoisomers of compounds, salts, prodrugs, or mixtures thereof, such as enantiomers and diastereomers (including all possible mixtures of stereoisomers). See, for example, WO 01 / 062726.

[0312] Furthermore, certain alkenyl-containing compounds may exist as Z (zusammen) or E (entgegen) isomers. In each case, this disclosure includes both mixtures and individual isomers.

[0313] Some compounds may also exist in tautomeric forms. Although not explicitly indicated in the formulas described herein, such forms are intended to be included within the scope of this disclosure.

[0314] A “prodrug” or “pharmaceutically acceptable prodrug” is a compound that, upon administration, is metabolized in a host, for example, by hydrolysis or oxidation, to form a compound of this disclosure (e.g., a compound of formula I or II). Typical examples of prodrugs include compounds having a biologically unstable or cleavable (protective) group on the functional moiety of an active compound. Prodrugs include compounds that can be oxidized, reduced, amination, deamination, hydroxylation, dehydroxylation, hydrolysis, dehydrolysis, alkylation, dealkylation, acylation, deacylation, phosphorylation, or dephosphorylation to produce an active compound. Examples of prodrugs using esters or aminophosphates as biologically unstable or cleavable (protective) groups are disclosed in U.S. Patents 6,875,751, 7,585,851, and 7,964,580, the disclosure of which is incorporated herein by reference. The prodrugs of this disclosure are metabolized to produce compounds of formula I or II. This disclosure includes prodrugs of the compounds described herein within its scope. For example, the routine procedure for selecting and preparing a suitable prodrug is described in “Design of Prodrugs” edited by H. Bundgaard, Elsevier, 1985.

[0315] As used herein, the phrase “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition, or medium suitable for the formulation of a medicine for medical or therapeutic use, such as a liquid or solid filter, diluent, excipient, solvent, or encapsulating material.

[0316] As used herein, the terms “logarithm of solubility,” “LogS,” or “logS” are used in the art to quantify the water solubility of a compound. The water solubility of a compound significantly affects its absorption and distribution characteristics. Low solubility is often accompanied by poor absorption. The LogS value is the unit stripping logarithm (base 10) of solubility measured in moles per liter.

[0317] Example

[0318] The invention will now be generally described, and will be more readily understood by referring to the following embodiments, which are included merely to illustrate certain aspects and embodiments of the invention and are not intended to limit the invention.

[0319] Example 1: Preparation of exemplary compounds

[0320]

[0321] To a dry DMF (6 mL) solution of indole-3-carboxaldehyde (2.8 mmol, 411 mg), Cu₂O (0.3 equivalence, 0.84 mmol, 120 mg), K₂CO₃ (2.0 equivalence, 5.6 mmol, 774 mg), and iodobenzene (2.0 equivalence, 5.6 mmol, 624 μL) were added sequentially. The reaction mixture was stirred and refluxed for 24 h, at which point TLC indicated completion. After cooling to 21 °C, the reaction mixture was filtered through a diatomaceous earth filter and eluted with ethyl acetate. The filtrate was washed with saturated NaCl solution, and the organic phase was dried over sodium sulfate and concentrated. The residue was purified by rapid column chromatography (hexane / EtOAc = 8:1) to obtain the desired product. Yield: 89%, 550.7 mg.

[0322] To a solution of 1-phenyl-indole-3-carboxaldehyde (1 mmol, 221 mg) in ethanol (1 mL), ethyl 2-cyanoacetate (1.3 equivalents, 1.3 mmol, 140 μL) and L-proline (40 mol%, 0.4 mmol, 58 mg) were added. The reaction mixture was stirred at 21 °C for 12 h, and a yellow solid gradually precipitated. After the reaction was complete, ice-cold water (2 mL) was added to the reaction flask. The solid was separated by filtration through a Buchner funnel, washed with water (2 mL × 3), and dried to obtain the desired product. Yield: 95%, 300 mg.

[0323] To a solution of (E)-2-cyano-3-(1-phenyl-1H-indol-3-yl)acrylate (0.32 mmol, 100 mg) in THF (2 mL), 0.5 N LiOH solution (3 equivalents, 0.6 mmol, 1.2 mL) was added. The reaction mixture was stirred at 21 °C for 1 h. After the reaction was complete as indicated by TLC, the THF was evaporated. Concentrated HCl was added dropwise to acidify the reaction mixture until the pH was below 1, and a yellow solid precipitated. Ice-cold water (5 mL) was added to the reaction mixture, and the solid was separated by filtration through a Buchner funnel and washed with water (5 mL × 3). After vacuum drying, the solid was washed 5 to 10 times with a solvent mixture of 2 mL (hexane / EtOAc = 5:1) and monitored by TLC until the nonpolar impurities disappeared (the nonpolar compounds were retrograde aldol condensation products and could be recovered from the filtrate). Finally, the purity of the product was checked by NMR. Yield: 65%, 60 mg.

[0324] (E)-2-Cyano-3-(1-Pheny-1H-Indole-3-yl)Acrylic Acid (JXL001 / UK5099)

[0325] 1 H NMR (500MHz, DMSO-d6) δ8.59(s,1H),8.56(s,1H),8.06(m,1H),7.65(m,4H),7.53(m,2H),7.34(m,2H).

[0326] 13 C NMR (126MHz, DMSO-d6) δ164.5,145.6,137.7,136.3,133.6,130.5,128.9,128.0,125.0,124.9,123.3,119.9,118.4,111.9,96.7.

[0327]

[0328] The following compounds were synthesized via a route similar to that described for JXL001: JXL002, JXL003, JXL004, JXL005, JXL006, JXL007, JXL012, JXL013, JXL014, JXL021, JXL025, JXL026, JXL027, JXL028, JXL029, JXL035, and JXL093.

[0329]

[0330] (E)-2-Cyano-3-(1H-Indole-3-yl)acrylic acid (JXL002)

[0331] 1 H NMR (500MHz, DMSO-d6) δ12.48(s,1H),8.51(s,1H),8.49(s,1H),7.91(d,J=6.5Hz,1H),7.53(d,J=7.0Hz,1H),7.23(m,2H).

[0332] 13 C NMR (126MHz, DMSO-d6) δ165.0,146.5,136.5,132.4,127.3,123.9,122.4,118.9,118.8,113.2,110.2,94.0.

[0333]

[0334] (E)-2-cyano-3-(1H-indol-3-yl)ethyl acrylate

[0335] 1 H NMR (500MHz, CDCl3) δ12.55(s,1H),8.53(s,1H),8.52(s,1H),7.92(d,J=7.6Hz,1H),7.53(d,J=7.8Hz,1 H), 7.26 (app.t, J = 7.4Hz, 1H), 7.22 (app.t, J = 7.4Hz, 1H), 4.24 (q, J = 7.0Hz, 2H), 1.26 (t, J = 7.0Hz, 3H).

[0336] 13 C NMR (126MHz, CDCl3) δ163.6,147.0,136.6,133.0,127.3,124.0,122.5,118.9,118.4,113.3,110.3,92.6,61.8,14.5.

[0337]

[0338] (E)-2-cyano-3-(1-phenyl-1H-indol-3-yl)ethyl acrylate (JXL004)

[0339] 1 H NMR (500MHz, CDCl3) δ8.71 (s, 1H), 8.66 (s, 1H), 7.90 (d, J = 7.2Hz, 1H), 7.54 (m, 6H), 7.36 (m, 2H), 4.39 (q, J = 7.1Hz, 2H), 1.42 (t, J = 7.1Hz, 3H).

[0340] 13C NMR (126MHz, CDCl3) δ163.6,145.6,137.8,136.4,133.2,129.9,128.5,124.8,124.4,123.0,118.5,117.9,111.6,111.5,95.4,62.0,14.3.

[0341]

[0342] (E)-3-(6-chloro-1-phenyl-1H-indol-3-yl)-2-cyanoacrylate (JXL005)

[0343] 1 H NMR (500MHz, CDCl3) δ8.67 (s, 1H), 8.58 (s, 1H), 7.81 (d, J = 8.5Hz, 1H), 7.60 (m, 2H) ,7.52(m,4H),7.34(d,J=8.4Hz,1H),4.39(q,J=7.1Hz,2H),1.41(t,J=7.1Hz,3H).

[0344] 13 C NMR (126MHz, CDCl3) δ163.4,145.1,137.3,136.8,133.5,130.5,130.0,128.8,126.8,124.8,123.6,119.5,117.5,111.6,111.4,96.4,62.1,14.2.

[0345]

[0346] (E)-2-cyano-3-(1-(2-methoxyphenyl)-1H-indol-3-yl)ethyl acrylate (JXL006)

[0347] 1 H NMR (500MHz, CDCl3) δ8.67(s,1H),8.66(s,1H),7.89(d,J=7.8Hz,1H),7.49(app.t,J=8.6Hz,1H),7.41(d,J=7.6Hz,1H),7.35(app.t ,J=7.3Hz,1H),7.30(t,J=7.5Hz,1H),7.23(d,J=8.1Hz,1H),7.13(m,2H),4.39(q,J=7.1Hz,2H),3.81(s,3H),1.41(t,J=7.1Hz,3H).

[0348] 13C NMR (126MHz, CDCl3) δ163.8,154.2,146.0,137.2,135.2,130.3,127.8,126.0,1 24.0,122.6,120.8,118.2,118.0,112.3,111.8,111.0,94.7,61.8,55.7,14.3.

[0349]

[0350] (E)-2-cyano-3-(1-(4-methoxyphenyl)-1H-indol-3-yl)ethyl acrylate (JXL007)

[0351] 1 H NMR (500MHz, CDCl3) δ8.65 (s, 1H), 8.64 (s, 1H), 7.89 (d, J = 7.2Hz, 1H), 7.44 (m, 3H), 7.35 (m,2H),7.07(d,J=8.8Hz,2H),4.38(q,J=7.1Hz,2H),3.90(s,3H),1.41(t,J=7.1Hz,3H).

[0352] 13 C NMR (126MHz, CDCl3) δ163.8,159.5,145.6,136.8,133.5,130.6,128.3,126. 2,124.2,122.9,118.4,118.0,115.0,111.5,111.1,94.9,61.9,55.6,14.2.

[0353]

[0354] (E)-2-cyano-3-(1-(4-methoxyphenyl)-1H-indol-3-yl)acrylic acid (JXL012)

[0355] 1 H NMR (500MHz, DMSO-d6) δ8.54 (s, 1H), 8.52 (s, 1H), 8.05 (d, J = 7.7Hz, 1H), 7.58 (app. d,J=8.7Hz,2H),7.44(m,1H),7.33(m,2H),7.16(app.d,J=8.7Hz,2H),3.82(s,3H).

[0356] 13C NMR (126MHz, DMSO-d6) δ164.7,160.5,136.7,133.9,130.5,127.8,126.6,124.8,123.2,120.0,119.5,118.6,115.6,115.4,111.9,110.9,55.9.

[0357]

[0358] (E)-3-(6-chloro-1-phenyl-1H-indol-3-yl)-2-cyanoacrylate (JXL013)

[0359] 1 H NMR (500MHz, DMSO-d6) δ13.58(br.s,1H),8.59(s,1H),8.54(s,1H),8.11(d,J=7.5Hz,1H),7.67(m,4H),7.53(m,2H),7.35(d,J=7.5Hz,1H).

[0360] 13 C NMR (126MHz, DMSO-d6) δ164.3,145.4,137.3,136.7,134.4,130.6,129.6,129.1,126.7,125.1,123.5,121.5,118.2,111.6,111.3,97.9.

[0361]

[0362] (E)-2-cyano-3-(1-(2-methoxyphenyl)-1H-indol-3-yl)acrylic acid (JXL014)

[0363] 1 H NMR (500MHz, DMSO-d6) δ8.55(s,1H),8.47(s,1H),8.02(d,J=7.4Hz,1H),7.54(m,2H),7.31(m,3H),7.15(m,2H),3.74(s,3H).

[0364] 13 C NMR (126MHz, DMSO-d6) δ164.6,154.2,145.8,137.3,135.1,131.1,128.2,127. 3,125.7,124.6,123.0,121.5,119.3,118.5,113.6,112.2,110.8,96.1,56.2.

[0365]

[0366] 2-((1-Phenylacetyl-1H-indol-3-yl)methylene)malononitrile (JXL021)

[0367] 1 H NMR (500MHz, CDCl3) δ8.62 (s, 1H), 8.13 (s, 1H), 7.80 (d, J = 7.4Hz, 1H), 7.61 (m, 2H), 7.53 (m, 4H), 7.40 (m, 2H).

[0368] 13 C NMR (126MHz, CDCl3) δ149.8,137.3,136.5,133.6,130.1,128.9,127.7,124.9,124.8,123.7,118.2,115.1,115.0,111.9,111.8,73.7.

[0369]

[0370] 2-(ethoxycarbonyl)-3-(1-phenyl-1H-indol-3-yl)acrylic acid (a mixture of E / Z isomers, 1:1 ratio) (JXL025)

[0371] 1 H NMR (500MHz, DMSO-d6) δ13.08(br.s,1H),7.87(m,3H),7.61(m,4H),7.52(m,2H),7.30(m,2H),4.26(m,2H),1.23(m,3H).

[0372] 13 C NMR (126MHz, DMSO-d6) δ168.9,167.8,166.1,164.9,138.3,136.1,132.9,132.1,131.1,130 .9,130.8,128.5,124.9,124.4,122.6,122.2,119.5,111.6,111.0,61.7,61.3,14.7,14.4.

[0373]

[0374] (E)-2-cyano-3-(4-fluoro-1-phenyl-1H-indole-3-yl)acrylic acid (JXL026)

[0375] 1H NMR (500MHz, DMSO-d6) δ8.65(s,1H),8.61(s,1H),7.67(m,4H),7.57(m,1H),7.36(m,2H),7.19(m,1H).

[0376] 13 C NMR(126MHz,DMSO-d6)δ164.4,156.8(d,J c-f =245.6Hz),146.4,138.7,137.6,133.4,130.7,129.5,125.9,125.4,118.2,116.3,116.2,109.5(d,J c-f =34.5Hz), 109.2(d,J c-f =23.2Hz), 98.0.

[0377]

[0378] (E)-2-cyano-3-(6-fluoro-1-phenyl-1H-indole-3-yl)acrylic acid (JXL027)

[0379] 1 H NMR (500MHz, DMSO-d6) δ13.59(br.s,1H),8.62(s,1H),8.59(s,1H),8.16(m,1H),7.66(m,4H),7.56(m,1H),7.36(d,J=9.2Hz,1H),7.25m,1H).

[0380] 13 C NMR(126MHz,DMSO-d6)δ164.6,160.8(d,J c-f =240.0Hz),145.7,137.6,136.6,134.4,130.8,129.1,125.0,124.7,121.6,118.4,111.8(d,J c-f =24.2Hz), 111.5, 98.7 (d, J) c-f =26.2Hz), 97.7.

[0381]

[0382] (E)-2-cyano-3-(7-fluoro-1-phenyl-1H-indole-3-yl)acrylic acid (JXL028)

[0383] 1H NMR (500MHz, DMSO-d6) δ13.62(br.s,1H),8.56(s,1H),8.47(s,1H),7.89(br.s,1H),7.61(m,5H),7.30(br.s,1H),7.17(br.s,1H).

[0384] 13 C NMR(126MHz,DMSO-d6)δ164.4,149.7(d,J c-f =247.5Hz),145.5,139.0,135.5,131.9,129.9,129.3,126.2,124.3,124.0,118.3,115.9,111.8,110.9(d,J c-f =17.4Hz), 98.1.

[0385]

[0386] (E)-2-cyano-3-(5-fluoro-1-phenyl-1H-indole-3-yl)acrylic acid (JXL029)

[0387] 1 H NMR (500MHz, DMSO-d6) δ13.57(br.s,1H),8.67(s,1H),8.60(s,1H),8.01(d,J=9.0Hz,1H),7.70(m,4H),7.58(m,2H),7.24(m,1H).

[0388] 13 C NMR(126MHz,DMSO-d6)δ164.6,159.4(d,J c-f =237.8Hz),145.8,137.7,135.1,133.1,130.7,129.2,129.1,125.2,118.5,113.7,113.2(d,J c-f =26.5Hz), 111.4, 105.7 (d, J) c-f =24.2Hz), 97.2.

[0389]

[0390] (E)-3-(4-chloro-1-phenyl-1H-indol-3-yl)-2-cyanoacrylate (JXL035)

[0391] 1H NMR (500MHz, DMSO-d6) δ9.22 (s, 1H), 8.68 (s, 1H), 7.64 (m, 3H), 7.59 (m, 2H), 7.48 (d, J = 7.6Hz, 1H), 7.38 (m, 1H), 7.32 (m, 1H).

[0392] 13 C NMR (126MHz, DMSO-d6) δ164.5,146.4,138.0,137.4,134.5,130.7,130.5,129.5,125.7,125.3,124.9,123.7,118.4,111.7,110.9,97.5.

[0393]

[0394] (E)-3-(4-chloro-1-phenyl-1H-indol-3-yl)-2-cyanoacrylate (JXL093)

[0395] 1 H NMR (500MHz, CDCl3) δ9.51 (s, 1H), 8.78 (s, 1H), 7.59 (m, 2H), 7.51 (m, 3H), 7.40 (dd, J = 8.3, 0.8Hz, 1H) ,7.32(dd,J=7.7,0.8Hz,1H),7.22(app.t,J=8.0Hz,1H),4.38(q,J=7.1Hz,2H),1.40(t,J=7.1Hz,3H).

[0396] 13 C NMR (126MHz, CDCl3) δ163.6,147.6,138.1,137.4,134.2,130.1,129.0,126.9,125.3,124.6,124.5,124.3,118.0,111.8,110.6,96.0,62.1,14.3.

[0397] Experimental details of JXL020 synthesis

[0398]

[0399] At 0 °C, 3,5-bis(trifluoromethyl)benzyl bromide (1.2 equivalents, 3.6 mmol, 660 μL) and KOH (1.2 equivalents, 3.6 mmol, 200 mg) were added to a dry DMF (6 mL) solution of indole-3-carboxaldehyde (3 mmol, 435 mg). The reaction mixture was stirred at 21 °C for 2 h. After the reaction was complete as indicated by TLC, water (6 mL) was added to a reaction vial. The reaction mixture was extracted with dichloromethane (15 mL × 3). The combined organic layers were dried over sodium sulfate and concentrated. The residue was purified by rapid column chromatography (hexane / EtOAc = 8:1) to give the desired product. Yield: 90%, 1001.7 mg.

[0400] To a solution of 1-(3,5-bis(trifluoromethyl)benzyl)-1H-indole-3-carboxaldehyde (1 mmol, 371 mg) in ethanol (1 mL), ethyl 2-cyanoacetate (1.3 equivalents, 1.3 mmol, 140 μL) and L-proline (40 mol%, 0.4 mmol, 58 mg) were added. The reaction mixture was stirred at 21 °C for 12 h, and a yellow solid gradually precipitated. After the reaction was complete, ice-cold water (2 mL) was added to the reaction flask. The solid was separated by filtration through a Buchner funnel, washed with water (2 mL × 3), and dried to obtain the desired product. Yield: 93%, 433 mg.

[0401] Add 0.5N LiOH solution (3 equivalents, 0.4 mmol, 0.8 mL) to a THF (2 mL) solution of (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyanoacrylate (0.21 mmol, 100 mg). Stir the reaction mixture at 21 °C for 1 h. After the reaction is complete as indicated by TLC, evaporate the THF. Acidify the reaction mixture by adding concentrated HCl dropwise until the pH is below 1, precipitating a yellow solid. Add ice-cold water (5 mL) to the reaction mixture, separate the solid by filtration through a Buchner funnel, and wash with water (5 mL × 3). After vacuum drying, wash the solid 5 to 10 times with a solvent mixture of 2 mL (hexane / EtOAc = 5:1) and monitor by TLC until the nonpolar impurities disappear (the nonpolar compounds are retrograde aldol condensation products and can be recovered from the filtrate). Finally, check the purity of the product by NMR. Yield: 55%, 52 mg.

[0402] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyanoacrylate (JXL020)

[0403] 1H NMR (500MHz, DMSO-d6) δ13.37(br.s,1H),8.75(s,1H),8.48(s,1H),7.99(m,4H),7.65(s,1H),7.28(m,2H),5.83(s,2H).

[0404] 13 C NMR(126MHz,DMSO-d6)δ164.7,145.7,140.3,136.3,134.8,131.1,130.8(q,J=31.1Hz),1 28.9,128.7,127.9,124.8,124.3,122.9(q,J=273.4Hz),122.2,119.3,118.3,95.6,49.2.

[0405] The following compounds were synthesized via a route similar to that described for JXL020: JXL008, JXL009, JXL010, JXL011, JXL015, JXL016, JXL017, JXL018, JXL019, JXL036, JXL037, JXL038, JXL039, JXL040, JXL041, JXL050, JXL051, JXL052, JXL053, JXL054, JXL0 55. JXL56, JXL057, JXL058, JXL059, JXL060, JXL061, JXL062, JXL063, JXL064, JXL065, JXL066, JXL068 , JXL069, JXL072, JXL073, JXL076, JXL077, JXL078, JXL081, JXL082, JXL087, JXL089, JXL090, JXL091.

[0406]

[0407] (E)-2-cyano-3-(1-(4-fluorobenzyl)-1H-indol-3-yl)ethyl acrylate (JXL008)

[0408] 1 H NMR(500MHz, CDCl3)δ8.60(app.s,2H),7.85(d,J=6.8Hz,1H),7.32(m,3H),7.15(m ,2H),7.03(app.t,2H),5.39(s,2H),4.37(q,J=7.1Hz,2H),1.40(t,J=7.1Hz,3H).

[0409] 13C NMR(126MHz,CDCl3)δ163.7,162.5(d,J c-f =247.7Hz),145.7,136.1,133.8,130.9,128.6,128.5,124.0,122.7,118.6,118.0,116.0(d,J c-f =21.9Hz),110.9,110.4,94.6,61.9,50.7,14.2.

[0410]

[0411] (E)-2-cyano-3-(1-(3,4-difluorobenzyl)-1H-indol-3-yl)ethyl acrylate (JXL009)

[0412] 1 H NMR (500MHz, CDCl3) δ8.60 (s, 1H), 8.59 (s, 1H), 7.86 (d, J = 7.8Hz, 1H), 7.33 (m, 2H), 7.28 (s, 1H), 7.13(m,1H),6.95(m,1H),6.89(m,1H),5.39(s,2H),4.38(q,J=7.1Hz,2H),1.40(t,J=7.1Hz,3H).

[0413] 13 C NMR (126MHz, CDCl3) δ163.7,150.7(dd,J=251.2,13.2Hz),150.2(dd,J=250.4,12.6Hz),145.7,136.1,133.7,132.3,128 .6,124.3,122.9,122.7,120.0,118.8,118.0(d,J=17.5Hz),115.9(d,J=18.0Hz),110.8,110.6,95.2,62.1,50.4,14.4.

[0414]

[0415] (E)-2-cyano-3-(1-(3,5-difluorobenzyl)-1H-indol-3-yl)ethyl acrylate (JXL010)

[0416] 1H NMR (500MHz, CDCl3) δ8.61(s,1H),8.59(s,1H),7.87(d,J=7.1Hz,1H),7.33(m,2H),7.26(d,J=7.2Hz,1H),6.7 5(app.t,J=8.7Hz,1H),6.64(app.d,J=5.7Hz,2H),5.41(s,2H),4.38(q,J=7.1Hz,2H),1.40(t,J=7.1Hz,3H).

[0417] 13 C NMR(126MHz,CDCl3)δ163.6,163.3(dd,J c-f =251.0,12.5Hz),145.5,139.2,136.0,133.6,128.4,124.3,122.9,118.7,118.0,110.7,110.6,109.5(dd,J c-f =19.9,6.4Hz),103.8(t,J) c-f =25.2Hz),95.3,62.0,50.4,14.2.

[0418]

[0419] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyanoacrylate (JXL011)

[0420] 1 H NMR (500MHz, CDCl3) δ8.62(s,1H),8.61(s,1H),7.90(d,J=7.7Hz,1H),7.85(s,1H),7.57(s,2H) ,7.35(m,2H),7.23(d,J=7.8Hz,1H),5.56(s,2H),4.38(q,J=7.1Hz,2H),1.40(t,J=7.1Hz,3H).

[0421] 13 C NMR (126MHz, CDCl3) δ163.6,145.6,138.1,136.0,133.4,132.7(q,J=33.8Hz),128.6,126.8,1 24.7,123.2,122.9(q,J=273.4Hz),122.6,119.0,118.0,111.2,110.4,95.9,62.1,50.5,14.3.

[0422]

[0423] (E)-2-cyano-3-(1-(4-fluorobenzyl)-1H-indol-3-yl)acrylic acid (JXL015)

[0424] 1 H NMR (500MHz, DMSO-d6) δ8.64(s,1H),8.46(s,1H),7.93(d,J=7.1Hz,1H),7.61(d,J=7.3Hz,1H),7.33(m,2H),7.26(m,2H),7.16(m,2H),5.60(s,2H).

[0425] 13 C NMR(126MHz,DMSO-d6)δ164.8,162.0(d,J c-f =244.3Hz),145.6,136.4,134.6,133.1,130.0,128.0,124.1,122.8,119.2,118.5,116.0(d,J c-f =21.7Hz),112.0,109.8,95.0,49.6.

[0426]

[0427] (E)-2-cyano-3-(1-(3,4-difluorobenzyl)-1H-indole-3-yl)acrylic acid (JXL016)

[0428] 1 H NMR(500MHz,DMSO-d6)δ13.34(br.s,1H),8.62(s,1H),8.47(s,1H),7.94(d,J=7.3Hz ,1H),7.61(d,J=7.6Hz,1H),7.40(m,2H),7.27(m,2H),7.10(br.s,1H),5.61(s,2H).

[0429] 13 C NMR(126MHz,DMSO-d6)δ164.8,149.7(dd,J c-f =253.3,13.6Hz),149.4(dd,J c-f =246.3,11.6Hz),145.7,136.4,134.7,134.5,128.0,124.7(dd,J c-f =5.9,3.0Hz),124.2,122.8,119.2,118.4,118.3(d,J c-f=17.0Hz), 117.1(d,J c-f =17.6Hz),112.0,109.9,95.0,49.3.

[0430]

[0431] (E)-2-cyano-3-(1-(3,5-difluorobenzyl)-1H-indole-3-yl)acrylic acid (JXL017)

[0432] 1 H NMR (500MHz, DMSO-d6) δ8.67 (s, 1H), 8.48 (s, 1H), 7.95 (d, J = 4.8Hz, 1H), 7. 59(d,J=4.4Hz,1H),7.27(m,2H),7.15(s,1H),6.98(br.s,2H),5.65(s,2H).

[0433] 13 C NMR(126MHz,DMSO-d6)δ164.8,162.9(dd,J c-f =247.0,12.8Hz),145.6,141.4,136.4,134.8,128.0,124.2,122.9,119.3,118.4,111.9,111.0(d,J c-f =26.1Hz), 110.8, 103.8(t,J) c-f =26.5Hz), 95.5, 49.5.

[0434]

[0435] (E)-3-(1-Benzyl-1H-indole-3-yl)-2-cyanoacrylate (JXL018)

[0436] 1 H NMR (500MHz, CDCl3) δ8.61 (s, 1H), 8.60 (s, 1H), 7.85 (d, J = 7.9Hz, 1H), 7.33 (m ,6H),7.17(m,2H),5.42(s,2H),4.37(q,J=7.1Hz,2H),1.40(t,J=7.1Hz,3H).

[0437] 13C NMR (126MHz, CDCl3) δ163.8,145.7,136.2,135.1,134.0,129.0,128.5,128. 2,126.8,124.0,122.6,118.5,118.1,111.0,110.3,94.3,61.8,51.4,14.2.

[0438]

[0439] (E)-3-(1-Benzyl-1H-indole-3-yl)-2-cyanoacrylate (JXL019)

[0440] 1 H NMR (500MHz, DMSO-d6) δ13.34(br.s,1H),8.65(s,1H),8.48(s,1H),7.93(d,J=6.9Hz,1H),7.60(d,J=6.8Hz,1H),7.25(m,7H),5.62(s,2H).

[0441] 13 C NMR (126MHz, DMSO-d6) δ164.8,145.7,136.8,136.5,134.7,129.1,128.2,127.6,124.0,122.7,120.0,119.2,118.5,112.1,109.7,94.7,50.4.

[0442]

[0443] (E)-2-cyano-3-(1-(3,4-difluorobenzyl)-5-fluoro-1H-indole-3-yl)acrylic acid (JXL036)

[0444] 1 H NMR (500MHz, DMSO-d6) δ8.69(s,1H),8.47(s,1H),7.84(d,J=9.6Hz,1H),7.63(dd,J=8.9,4.3Hz,1H),7.41(m,2H),7.14(m,2H),5.61(s,2H).

[0445] 13 C NMR(126MHz,DMSO-d6)δ164.9,159.4(d,J c-f =237.3Hz), 149.8(dd,J c-f =247.1,12.7Hz),149.5(dd,J c-f=246.6,12.3Hz),146.0,136.1,134.5,133.1,129.1,125.0,118.5,118.4,117.4,113.6,112.5(d,J c-f =26.2Hz), 110.1, 105.2 (d, J) c-f =25.1Hz), 95.5, 49.7.

[0446]

[0447] (E)-2-cyano-3-(1-(3,5-difluorobenzyl)-5-fluoro-1H-indole-3-yl)acrylic acid (JXL037)

[0448] 1 H NMR(500MHz,DMSO-d6)δ8.71(s,1H),8.48(s,1H),7.85(dd,J=9.6,2.0Hz,1H),7 .62(dd,J=8.9,4.3Hz,1H),7.16(m,2H),7.00(app.d,J=6.2Hz,2H),5.65(s,2H).

[0449] 13 C NMR(126MHz,DMSO-d6)δ164.9,163.0(d,J c-f =247.7Hz), 159.3(d,J) c-f =237.6Hz),145.9,141.4,136.3,133.2,129.1,118.5,113.6,112.6(d,J c-f =26.3Hz),111.3,110.2,105.3(d,J c-f =24.9Hz), 104.0(t,J c-f =25.2Hz), 95.9, 49.8.

[0450]

[0451] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-5-fluoro-1H-indole-3-yl)-2-cyanoacrylate (JXL038)

[0452] 1H NMR(500MHz,DMSO-d6)δ8.80(s,1H),8.49(s,1H),8.06(s,1H),8.04(s,2H),7.86(dd,J =9.6, 2.1Hz, 1H), 7.69 (dd, J = 8.9, 4.3Hz, 1H), 7.17 (dt, J = 9.0, 2.2Hz, 1H), 5.83 (s, 2H).

[0453] 13 C NMR(126MHz,DMSO-d6)δ164.8,159.3(d,J c-f =237.5Hz),145.9,140.3,136.2,133.1,131.1(q,J c-f =33.1Hz), 129.1, 123.6(q,J) c-f =272.2Hz),118.4,113.5,112.7,112.5,110.4,105.5,105.3,96.1,49.6.

[0454]

[0455] (E)-2-cyano-3-(1-(3,4-difluorobenzyl)-6-fluoro-1H-indole-3-yl)acrylic acid (JXL039)

[0456] 1 H NMR (500MHz, DMSO-d6) δ8.64(s,1H),8.46(s,1H),7.99(dd,J=8.7,5.1Hz,1H),7.58(dd,J=9.8,1.8Hz,1H),7.43(m,2H),7.12(m,2H),5.57(s,2H).

[0457] 13 C NMR(126MHz,DMSO-d6)δ164.8,160.3(d,J c-f =239.3Hz), 149.8(dd,J c-f =239.3,25.2Hz),149.6(dd,J c-f =246.3,25.2Hz),145.8,136.8,135.3,134.5,125.1,125.0,124.6,121.1,118.5,117.5,111.3(d,J c-f =23.9Hz), 110.2, 98.7 (d, J) c-f =26.5Hz), 96.2, 49.4.

[0458]

[0459] (E)-2-cyano-3-(1-(3,5-difluorobenzyl)-6-fluoro-1H-indole-3-yl)acrylic acid (JXL040)

[0460] 1 H NMR(500MHz,DMSO-d6)δ8.65(s,1H),8.46(s,1H),8.00(dd,J=8.6,5.2Hz,1H ),7.57(d,J=9.7Hz,1H),7.15(m,2H),7.03(s,1H),7.02(s,1H),5.62(s,2H).

[0461] 13 C NMR(126MHz,DMSO-d6)δ164.8,163.0(d,J c-f =239.4Hz), 162.9(d,J) c-f =248.6Hz), 160.4(d,J) c-f =239.4Hz),145.5,141.3,136.8,135.2,124.6,121.1,118.4,111.3,111.1,110.3,104.0(t,J c-f =25.2Hz), 98.7(d,J c-f =26.5Hz), 96.9, 49.6.

[0462]

[0463] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-6-fluoro-1H-indole-3-yl)-2-cyanoacrylate (JXL041)

[0464] 1 H NMR(500MHz,DMSO-d6)δ8.74(s,1H),8.48(s,1H),8.06(app.s,3H),8.01(dd,J=8.7 ,5.1Hz,1H),7.66(dd,J=9.8,2.0Hz,1H),7.13(dt,J=9.3,2.1Hz,1H),5.78(s,2H).

[0465] 13 C NMR(126MHz,DMSO-d6)δ164.7,159.9(d,J c-f =264.6Hz),145.8,140.2,136.8,135.3,131.1(q,Jc-f =33.3Hz),129.1,124.5,123.6(q,J c-f =273.7Hz),122.5,121.2,118.4,111.4(d,J c-f =25.2Hz), 110.4, 98.6 (d, J) c-f =27.2Hz), 96.7, 49.4.

[0466]

[0467] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-4-chloro-1H-indol-3-yl)-2-cyanoacrylate (JXL050)

[0468] 1 H NMR (500MHz, DMSO-d6) δ9.20 (s, 1H), 8.90 (s, 1H), 8.06 (m, 3H), 7.72 (d, J = 7.5Hz, 1H), 7.32 (m, 2H), 5.86 (s, 2H).

[0469] 13 C NMR(126MHz,DMSO-d6)δ164.7,146.7,140.0,138.0,135.6,131.1(q,J c-f =33.1Hz),129.1,125.6,125.1,124.7,124.5,123.7,123.6(q,J c-f =273.7Hz),122.5,111.6,109.9,96.7,49.6.

[0470]

[0471] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-4-bromo-1H-indole-3-yl)-2-cyanoacrylate (JXL051)

[0472] 1 H NMR (500MHz, DMSO-d6) δ9.37(s,1H),8.91(s,1H),8.07(app.s,3H),7.77(d,J=8.2Hz,1H),7.51(d,J=7.6Hz,1H),7.23(t,J=8.0Hz,1H),5.85(s,2H).

[0473] 13C NMR(126MHz,DMSO-d6)δ164.7,146.3,140.0,138.0,135.8,131.0(q,J c-f =33.1Hz),129.1,127.9,125.4,124.9,123.7(q,J c-f =273.2Hz),122.5,118.2,113.6,122.1,110.2,96.6,49.5.

[0474]

[0475] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylate (JXL052)

[0476] 1 H NMR(500MHz,DMSO-d6)δ8.80(s,1H),8.58(s,1H),8.05(app.s,3H),7.54(d,J= 8.2Hz, 1H), 7.29 (d, J=12.9Hz, 1H), 7.09 (dd, J=11.1, 8.2Hz, 1H), 5.85 (s, 2H).

[0477] 13 C NMR(126MHz,DMSO-d6)δ164.5,155.7(d,J c-f =239.4Hz),146.5,140.1,138.8,134.5,131.1(q,J c-f =33.0Hz), 129.0, 125.1 (d, J) c-f =7.6Hz), 124.7, 123.6(q,J) c-f =273.5Hz),122.5,118.1,116.1(d,J c-f =18.5Hz),108.8,108.5,97.2,49.7.

[0478]

[0479] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-7-fluoro-1H-indole-3-yl)-2-cyanoacrylate (JXL053)

[0480] 1H NMR(500MHz,DMSO-d6)δ8.73(br.s,1H),8.48(br.s,1H),8.06(br.s,1H),7.89(b r.s,2H),7.78(br.s,J=7.4Hz,1H),7.22(br.s,1H),7.10(br.s,1H),5.89(s,2H).

[0481] 13 C NMR(126MHz,DMSO-d6)δ164.6,159.7(d,J c-f =245.7Hz),145.6,141.0,136.2,132.1,130.0(q,J c-f =33.0Hz),128.4,123.9,123.8,123.6(q,J c-f =273.3Hz), 122.4(d,J c-f =18.9Hz),118.1,115.8,110.6,110.2(d,J c-f =18.9Hz), 97.3, 52.0.

[0482]

[0483] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-5-chloro-1H-indol-3-yl)-2-cyanoacrylate (JXL054)

[0484] 1 H NMR (500MHz, DMSO-d6) δ8.78(s,1H),8.51(s,1H),8.12(s,1H),8.06(s,1H),8.03(s,2H),7.70(d,J=7.4Hz,1H),7.33(d,J=7.0Hz,1H),5.83(s,2H).

[0485] 13 C NMR(126MHz,DMSO-d6)δ164.7,145.7,140.2,135.9,135.1,131.1(q,J c-f =33.0Hz),129.3,129.0,127.9,124.4,123.6(q,J c-f =274.0Hz),122.5,119.5,118.4,113.6,109.9,96.9,49.5.

[0486]

[0487] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-4-cyano-1H-indole-3-yl)-2-cyanoacrylate (JXL055)

[0488] 1 H NMR (500MHz, DMSO-d6) δ9.03 (s, 1H), 9.00 (s, 1H), 8.09 (m, 4H), 7.80 (d, J = 7.3Hz, 1H), 7.47 (t, J = 7.8Hz, 1H), 5.90 (s, 2H).

[0489] 13 C NMR(126MHz,DMSO-d6)δ164.4,144.4,139.9,136.7,131.1(q,J c-f =33.0Hz),129.7,129.2,126.6,124.3,123.6(q,J c-f =273.7Hz),122.7,118.6,117.9,117.8,109.3,101.7,98.1,49.5.

[0490]

[0491] (E)-1-(3,5-bis(trifluoromethyl)benzyl)-3-(2-carboxy-2-cyanovinyl)-1H-indole-4-carboxylic acid (JXL056)

[0492] 1 H NMR(500MHz,DMSO-d6)δ9.26(br.s,1H),8.88(br.s,1H),8.05(app.s,3H),7.94(br.s,1H),7.76(br.s,1H),7.37(br.s,1H),5.85(s,2H).

[0493] 13 C NMR(126MHz,DMSO-d6)δ169.3,165.0,150.6,140.2,137.7,136.0,131.1(q,J c-f =33.0Hz),129.0,125.9,125.2,124.7,123.6(q,J c-f =273.7Hz),123.5,122.5,118.3,116.1,110.1,96.4,49.3.

[0494]

[0495] (E)-3-(4-(benzyloxy)-1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyanoacrylate (JXL057)

[0496] 1 H NMR(500MHz,DMSO-d6)δ9.15(s,1H),8.72(s,1H),8.04(s,1H),7.99(s,2H),7.54 (br.s,3H),7.37(br.s,2H),7.28(m,2H),6.95(s,1H),5.81(s,2H),5.28(s,2H).

[0497] 13 C NMR(126MHz,DMSO-d6)δ165.0,153.7,148.6,140.5,137.9,133.4,131.2(q,J c-f =32.8Hz),128.9,128.8,128.1,127.5,125.4,124.7,123.6(q,J c-f =273.7Hz),122.3,118.5,117.0,110.5,105.6,105.2,95.5,70.0,49.5.

[0498]

[0499] (E)-3-(6-(benzyloxy)-1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyanoacrylate (JXL058)

[0500] 1 H NMR(500MHz,DMSO-d6)δ8.63(s,1H),8.43(s,1H),8.05(s,1H),8.03(s,2H),7.8 6(d,J=8.5Hz,1H),7.32(m,6H),6.97(d,J=8.3Hz,1H),5.77(s,2H),5.09(s,2H).

[0501] 13 C NMR (126MHz, DMSO-d6) δ165.0,156.7,146.1,140.5,137.5,137.3,134.3,131.0(q,J c-f =32.8Hz),128.9,128.8,128.3,128.2,126.9,123.6(q,J c-f=273.4Hz),122.4,121.9,118.5,113.3,110.5,96.8,95.6,70.2,49.2.

[0502]

[0503] (E)-3-(7-(benzyloxy)-1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyanoacrylate (JXL059)

[0504] 1 H NMR(500MHz,DMSO-d6)δ8.61(br.s,1H),8.45(br.s,1H),7.97(br.s,1H),7.60(br.s,2H),7 .51(br.s,1H),7.25(br.s,2H),7.16(br.s,2H),6.91(br.s,1H),5.94(s,2H),5.13(s,2H).

[0505] 13 C NMR (126MHz, DMSO-d6) δ164.8,146.6,145.7,142.3,136.6,135.4,130.7(q,J c-f =32.8Hz),130.5,128.8,128.4,128.0,127.5,125.7,124.1,123.6(q,J c-f =273.7Hz),121.8,118.3,111.8,110.3,107.1,96.2,70.3,52.4.

[0506]

[0507] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-4-methoxy-1H-indol-3-yl)-2-cyanoacrylate (JXL060)

[0508] 1 H NMR (500MHz, DMSO-d6) δ8.99(s,1H),8.71(s,1H),8.05(s,1H),8.00(s,2H),7.24(app.s,2H),6.82(d,J=6.0Hz,1H),5.81(s,2H),3.92(s,3H).

[0509] 13C NMR(126MHz,DMSO-d6)δ165.0,154.8,148.6,140.5,137.8,133.3,131.0(q,J c-f =32.8Hz),128.9,125.4,123.6(q,J c-f =273.7Hz),122.5,118.5,116.8,110.5,105.0,104.3,95.3,56.2,49.5.

[0510]

[0511] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-5-bromo-1H-indol-3-yl)-2-cyanoacrylate (JXL061)

[0512] 1 H NMR (500MHz, DMSO-d6) δ8.75(s,1H),8.49(s,1H),8.25(s,1H),8.06(s,1H),8.02(m,3H),7.64(app.s,1H),7.44(app.s,1H),5.82(s,2H).

[0513] 13 C NMR(126MHz,DMSO-d6)δ164.7,145.3,140.3,135.5,135.3,131.1(q,J c-f =32.8Hz),129.8,128.9,127.0,124.7,123.6(q,J c-f =274.0Hz),122.5,122.3,118.5,113.9,109.9,97.7,49.5.

[0514]

[0515] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-6-bromo-1H-indole-3-yl)-2-cyanoacrylate (JXL062)

[0516] 1 H NMR (500MHz, DMSO-d6) δ8.72(s,1H),8.47(s,1H),8.06(s,2H),8.03(s,1H),7.94(d,J=8.2Hz,1H),7.39(d,J=7.9Hz,1H),5.81(s,2H).

[0517] 13C NMR(126MHz,DMSO-d6)δ164.7,145.6,140.2,137.4,135.3,131.1(q,J c-f =32.8Hz),129.1,127.0,125.9,123.6(q,J c-f =274.0Hz),122.5,121.5,118.2,117.2,114.8,110.3,96.9,49.3.

[0518]

[0519] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-6-chloro-1H-indol-3-yl)-2-cyanoacrylate (JXL063)

[0520] 1 H NMR (500MHz, DMSO-d6) δ8.75(s,1H),8.47(s,1H),8.05(m,3H),8.00(d,J=7.4Hz,1H),7.89(s,1H),7.28(d,J=6.5Hz,1H),5.81(s,2H).

[0521] 13 C NMR(126MHz,DMSO-d6)δ164.7,145.6,140.2,137.0,135.4,131.1(q,J c-f =32.8Hz),129.2,129.0,126.7,123.6(q,J c-f =274.0Hz),123.3,122.5,121.2,118.2,111.9,110.3,97.0,49.3.

[0522]

[0523] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-7-chloro-1H-indol-3-yl)-2-cyanoacrylate (JXL064)

[0524] 1 H NMR (500MHz, DMSO-d6) δ8.69(s,1H),8.50(s,1H),8.03(s,1H),7.97(d,J=7.5Hz,1H),7.73(s,2H),7.32(d,J=7.1Hz,1H),7.25(m,1H),6.14(s,2H).

[0525] 13C NMR(126MHz,DMSO-d6)δ164.5,145.1,142.1,136.9,131.5,131.2(q,J c-f =32.8Hz),127.6,126.1,124.1,123.6(q,J c-f =274.0Hz),122.5,122.0,118.8,118.0,116.9,110.3,97.9,51.8.

[0526]

[0527] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-4-bromo-1H-indole-3-yl)-2-cyanoacrylate (JXL065)

[0528] 1 H NMR(500MHz,DMSO-d6)δ9.67(s,1H),8.71(s,1H),7.86(s,1H),7.56(s,2H),7.52(d, J=7.4Hz,1H),7.17(m,2H),5.55(s,2H),4.37(q,J=7.1Hz,2H),1.40(t,J=7.1Hz,3H).

[0529] 13 C NMR(126MHz,DMSO-d6)δ163.4,146.9,137.6,137.4,134.3,132.7(q,J c-f =33.9Hz),128.2,126.7,125.6,125.1,122.8(q,J c-f =273.4Hz),122.7,118.0,114.9,111.8,109.9,96.3,62.2,50.6,14.3.

[0530]

[0531] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-4-fluoro-1H-indole-3-yl)-2-cyanoacrylate (JXL066)

[0532] 1H NMR(500MHz,DMSO-d6)δ8.86(s,1H),8.61(s,1H),7.86(s,1H),7.56(s,2H),7.2 3(m,1H),7.01(m,2H),5.54(s,2H),4.37(q,J=7.1Hz,2H),1.39(t,J=7.1Hz,3H).

[0533] 13 C NMR (126MHz, DMSO-d6) δ163.1,157.4(d,J=243.9Hz),147.4,138.2(d,J=10.1Hz),137.8,133.2,132.7(q,J c-f =33.9Hz),126.7,125.3,122.8(q,J c-f =273.4Hz),122.7,117.9,116.9(d,J c-f =17.6Hz), 110.0, 109.0 (d, J) c-f =19.5Hz),106.6,97.1,62.2,50.7,14.3.

[0534]

[0535] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyanoacrylate tert-butyl ester (JXL068)

[0536] 1 H NMR (500MHz, CDCl3) δ8.57 (s, 1H), 8.54 (s, 1H), 7.86 (d, J = 7.6Hz, 1H), 7.84 (s, 1H),7.56(s,2H),7.33(m,2H),7.22(d,J=7.4Hz,1H),5.55(s,2H),1.59(s,9H).

[0537] 13 C NMR (126MHz, CDCl3) δ162.4,144.7,138.2,135.9,133.0,132.7(q,J c-f =32.8Hz),128.6,126.7,124.5,123.0,122.9(q,J c-f =277.2Hz),122.6,119.0,118.2,111.1,110.4,97.7,82.9,50.4,28.1.

[0538]

[0539] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-cyanoacrylate (JXL069)

[0540] 1 H NMR (500MHz, DMSO-d6) δ8.85(s,1H),8.47(m,3H),8.09(s,2H),8.04(s,1H),7.35(dd,J=7.1,4.6Hz,1H),5.84(s,2H).

[0541] 13 C NMR(126MHz,DMSO-d6)δ185.9,164.6,147.8,146.1,145.5,141.0,135.2,131.0(q,J c-f =32.8Hz),129.4,126.9,123.6(q,J c-f =274.0Hz),122.3,120.1,119.1,118.2,97.1,47.8.

[0542]

[0543] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-pyrrolo[3,2-b]pyridin-3-yl)-2-cyanoacrylate (JXL072)

[0544] 1 H NMR (500MHz, DMSO-d6) δ8.94(s,1H),8.68(s,1H),8.54(s,1H),8.11(m,4H),7.35(s,1H),5.87(s,2H).

[0545] 13 C NMR(126MHz,DMSO-d6)δ164.5,145.9,144.2,140.1,135.9,131.1(q,J c-f =32.8Hz),129.6,129.2,124.7,123.6(q,J c-f =274.0Hz),122.5,120.0,119.5,117.9,110.1,97.2,49.8.

[0546]

[0547] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-pyrrolo[2,3-c]pyridin-3-yl)-2-cyanoacrylate (JXL073)

[0548] 1 H NMR (500MHz, DMSO-d6) δ9.14(s,1H),8.96(s,1H),8.55(s,1H),8.40(d,J=5.1Hz,1H),8.19(s,2H),8.11(app.s,2H),5.95(s,2H).

[0549] 13 C NMR (126MHz, DMSO-d6) δ164.5,145.5,140.7,139.9,138.1,134.5,133.6,131.1(q,J c-f =32.8Hz),129.4,127.2,123.6(q,J c-f =274.0Hz),122.7,118.0,114.6,109.7,97.9,49.8.

[0550]

[0551] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-4-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-cyanoacrylate (JXL076)

[0552] 1 H NMR (500MHz, DMSO-d6) δ9.00(s,1H),8.98(s,1H),8.36(s,1H),8.12(s,2H),8.04(br.s,1H),7.47(br.s,1H),5.85(s,2H).

[0553] 13 C NMR (126MHz, DMSO-d6) δ164.4,148.7,145.8,145.7,140.2,136.1,135.3,131.0(q,J c-f =32.8Hz), 129.5, 123.6(q,J) c-f =274.0Hz),122.4,120.4,117.7,116.8,108.2,98.1,48.2.

[0554]

[0555] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-4-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-cyanoacrylate (JXL077)

[0556] 1 H NMR (500MHz, DMSO-d6) δ9.17(s,1H),9.02(s,1H),8.27(d,J=5.0Hz,1H),8.12(s,2H),8.05(s,1H),7.63(d,J=5.0Hz,1H),5.85(s,2H).

[0557] 13 C NMR(126MHz,DMSO-d6)δ164.4,148.2,145.5,145.3,140.2,135.4,131.0(q,J c-f =32.8Hz),129.6,124.9,123.7,123.6(q,J c-f =274.0Hz),122.4,118.4,117.7,108.6,97.9,48.2.

[0558]

[0559] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyanoacrylate (JXL078)

[0560] 1 H NMR (500MHz, CDCl3) δ8.62 (s, 1H), 8.61 (s, 1H), 7.89 (d, J = 7.7Hz, 1H), 7.8 4(s,1H),7.57(s,2H),7.35(m,2H),7.24(m,1H),5.55(s,2H),3.92(s,3H).

[0561] 13 C NMR (126MHz, CDCl3) δ164.5,145.8,138.1,136.0,133.5,132.7(q,J c-f =32.8Hz),128.6,126.8,124.7,123.9,123.6(q,J c-f =274.0Hz),123.2,119.1,118.0,111.2,110.5,95.4,53.0,50.5.

[0562]

[0563] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-(morpholin-4-carbonyl)acrylonitrile (JXL081)

[0564] 1 H NMR (500MHz, CDCl3) δ8.48(s,1H),8.34(s,1H),7.84(s,2H),7.56(app.s,2H),7.32(m,2H),7.22(m,1H),5.54(s,2H),3.77(br.s,8H).

[0565] 13 C NMR (126MHz, CDCl3) δ164.0,145.2,138.4,135.8,132.7(q,J c-f =34.0Hz),131.8,128.4,126.8,125.0,124.5,123.1(q,J c-f =273.3Hz),122.8,121.8,119.0,118.6,111.4,110.3,98.1,66.7,50.3,50.0.

[0566]

[0567] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-cyanoacrylate (JXL082)

[0568] 1 H NMR (500MHz, CDCl3) δ8.64(s,1H),8.51(s,1H),8.48(dd,J=4.6,1.2Hz,1H),8.21(dd,J=7.9,1.2Hz,1H),7.8 3(s,1H),7.77(s,2H),7.34(dd,J=7.9,4.6Hz,1H),5.69(s,2H),4.38(q,J=7.1Hz,2H),1.40(t,J=7.1Hz,3H).

[0569] 13 C NMR (126MHz, CDCl3) δ163.3,147.6,145.7,145.0,138.5,132.7,132.3(q,J c-f =33.6Hz),127.9,127.7,123.1(q,J c-f=273.3Hz),122.5,120.3,119.0,117.6,109.4,97.0,62.3,48.3,14.3.

[0570]

[0571] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-4-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-cyanoacrylate (JXL087)

[0572] 1 H NMR (500MHz, CDCl3) δ9.22 (s, 1H), 8.77 (s, 1H), 8.32 (d, J = 5.2Hz, 1H), 7.83 (s, 1H), 7.77 (s,2H),7.31(d,J=5.2Hz,1H),5.67(s,2H),4.37(q,J=7.1Hz,2H),1.39(t,J=7.1Hz,3H).

[0573] 13 C NMR (126MHz, CDCl3) δ163.0,148.5,146.2,145.4,138.1,137.4,133.1,132.4(q,J c-f =33.7Hz), 128.0, 122.9(q,J) c-f =273.4Hz),122.6,121.8,117.6,117.2,109.7,97.8,62.4,48.6,14.3.

[0574]

[0575] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-4-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-cyanoacrylate (JXL088)

[0576] 1 H NMR (500MHz, CDCl3) δ9.40 (s, 1H), 8.78 (s, 1H), 8.22 (d, J = 5.1Hz, 1H), 7.83 (s, 1H), 7.77 (s,2H),7.49(d,J=5.1Hz,1H),5.67(s,2H),4.37(q,J=7.1Hz,2H),1.39(t,J=7.1Hz,3H).

[0577] 13C NMR (126MHz, CDCl3) δ163.1,148.1,145.7,145.1,138.1,137.4,133.4,132.4(q,J c-f =33.7Hz), 123.7, 122.9(q,J) c-f =273.4Hz),122.6,119.7,117.6,110.0,101.4,97.4,62.4,48.6,14.3.

[0578]

[0579] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-5-chloro-1H-indol-3-yl)-2-cyanoacrylate (JXL089)

[0580] 1 H NMR(500MHz, CDCl3)δ8.60(s,1H),8.50(s,1H),7.85(s,1H),7.83(s,1H),7.55(s,2H),7.28(dd,J =8.7,1.7Hz,1H),7.14(d,J=8.7Hz,1H),5.54(s,2H),4.38(q,J=7.1Hz,2H),1.40(t,J=7.1Hz,3H).

[0581] 13 C NMR (126MHz, CDCl3) δ163.2,144.8,137.7,134.3,134.1,132.8(q,J c-f =33.7Hz),129.7,129.3,126.7,125.1,122.8(q,J c-f =273.4Hz),122.7,118.9,117.7,111.6,110.6,96.9,62.3,50.6,14.3.

[0582]

[0583] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-4-cyano-1H-indole-3-yl)-2-cyanoacrylate (JXL090)

[0584] 1H NMR (500MHz, CDCl3) δ9.29 (s, 1H), 8.78 (s, 1H), 7.87 (s, 1H), 7.68 (dd, J = 7.4, 0.8Hz, 1H), 7.56 (s, 2H), 7.49 ( dd,J=8.4,0.8Hz,1H),7.38(dd,J=8.4,7.4Hz,1H),5.62(s,2H),4.38(q,J=7.1Hz,2H),1.40(t,J=7.1Hz,3H).

[0585] 13 C NMR (126MHz, CDCl3) δ162.6,144.3,137.3,136.2,135.1,132.9(q,J c-f =33.7Hz),129.4,127.3,126.7,124.1,123.0,122.8(q,J c-f =273.4Hz),117.9,117.5,115.4,110.7,103.5,98.6,62.4,50.6,14.3.

[0586]

[0587] (E)-1-(3,5-bis(trifluoromethyl)benzyl)-3-(2-cyano-3-ethoxy-3-oxoprop-1-en-1-yl)-1H-indole-4-carboxylic acid methyl ester (JXL091)

[0588] 1 H NMR (500MHz, CDCl3) δ9.36 (s, 1H), 8.71 (s, 1H), 7.93 (dd, 1H, J = 7.4, 1.1Hz, 1H), 7.85 (s, 1H), 7.55 (s, 2H), 7.40 (dd, J =8.3,1.1Hz,1H),7.34(dd,J=8.3,7.4Hz,1H),5.58(s,2H),4.37(q,J=7.1Hz,2H),4.04(s,3H),1.40(t,J=7.1Hz,3H).

[0589] 13 C NMR (126MHz, CDCl3) δ167.8,163.5,150.1,137.8,137.2,134.9,132.8(q,J c-f =33.7Hz),126.7,125.8,125.1,123.6,122.8(q,J c-f=273.4Hz),122.7,121.7,118.0,114.6,111.3,96.4,62.1,52.6,50.5,14.3.

[0590] Experimental details of JXL024 synthesis

[0591]

[0592] Thiazolidine-2,4-dione (1 equivalent, 0.4 mmol, 46.8 mg) and NaOAc (3 equivalents, 98 mg) were added to a solution of 1-phenyl-1H-indole-3-carboxaldehyde (0.4 mmol, 90 mg) in AcOH (3 mL). The reaction mixture was refluxed and stirred for 24 hours. After cooling to 21 °C, the reaction mixture was filtered through a vacuum filter and washed with AcOH (3 mL × 3) and water (5 mL × 3). After vacuum drying, the desired product was obtained. Yield: 34%, 44 mg.

[0593] (Z)-5-((1-Phenylacetyl-1H-indol-3-yl)methylene)thiazolidin-2,4-dione (JXL024)

[0594] 1 H NMR(500MHz DMSO-d6) δ7.98(m,2H),7.79(s,1H),7.66(app.d,J=7.7Hz,2H),7.62(app.t,J=7.7Hz,2H),7.54(d,J=8.0Hz,1H),7.49(t,J=7.1Hz,1H),7.30(m,2H).

[0595] 13 C NMR (126MHz, DMSO-d6) δ172.5,169.5,138.4,136.2,130.5,129.9,128.3,128.2,125.0,124.6,122.4,121.5,121.4,119.6,113.0,111.5.

[0596] The following compounds were synthesized via a route similar to that described for JXL024: JXL067, JXL070, JXL072, JXL074, and JXL075.

[0597]

[0598] (Z)-2-Imine-5-((1-Pheny-1H-indol-3-yl)methylene)thiazolidin-4-one (JXL023)

[0599] 1H NMR (500MHz, DMSO-d6) δ11.94(br.s,1H),9.33(br.s,1H),8.97(s,1H),7.64(m,10H).

[0600] 13 C NMR (126MHz, DMSO-d6) δ180.9,174.8,172.5,138.6,136.2,130.5,129.3,128.1,126.3,124.8,124.5,122.2,120.3,119.7,113.5,111.5.

[0601]

[0602] (Z)-5-((1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)methylene)-2-iminothiazolidin-4-one (JXL067)

[0603] 1 H NMR(500MHz,DMSO-d6)δ9.21(s,1H),9.01(s,1H),8.03(s,1H),7.94(br.s,3H),7.86(d, J=7.7Hz,1H),7.81(s,1H),7.58(d,J=7.9Hz,1H),7.24(m,1H),7.19(m,1H),5.76(s,2H).

[0604] 13 C NMR(126MHz,DMSO-d6)δ180.0,174.9,141.4,136.5,131.0(q,J c-f =32.8Hz),130.4,128.5,127.9,125.1,123.8,123.6(q,J c-f =274.0Hz),122.1,121.7,120.8,119.3,111.8,111.3,48.9.

[0605]

[0606] (Z)-5-((4-fluoro-1-phenyl-1H-indol-3-yl)methylene)thiazolidin-2,4-dione (JXL070)

[0607] 1H NMR (500MHz, DMSO-d6) δ12.45(br.s,1H),8.11(s,1H),7.83(s,1H),7.65(m,4H),7.52(s,1H),7.32(m,2H),7.12(m,1H).

[0608] 13 C NMR(126MHz,DMSO-d6)δ168.0,167.7,156.9(d,J c-f =245.7Hz), 138.7(d,J) c-f =10.1Hz),137.9,130.8,130.5,128.8,125.5,125.4,124.3,120.0,116.2(d,J c-f =18.9Hz),110.7,108.5,108.1(d,J c-f =18.9Hz).

[0609]

[0610] (Z)-5-((6-fluoro-1-phenyl-1H-indol-3-yl)methylene)thiazolidin-2,4-dione (JXL071)

[0611] 1 H NMR(500MHz,DMSO-d6)δ12.42(s,1H),8.06(app.s,2H),7.85(s,1H),7.67(m ,2H),7.62(m,2H),7.50(m,1H),7.31(d,J=9.6Hz,1H),7.16(t,J=8.5Hz,1H).

[0612] 13 C NMR(126MHz,DMSO-d6)δ168.0,167.7,160.6(d,J c-f =245.7Hz), 138.0, 136.3 (d, J) c-f =12.6Hz),131.0,130.6,128.5,125.0,124.8,123.5,121.3(d,J c-f =10.0Hz),120.0,112.6,111.1(d,J c-f =18.9Hz), 98.2(d,J c-f =18.9Hz).

[0613]

[0614] (Z)-5-Benzylthiazolidin-2,4-dione (JXL074)

[0615] 1 H NMR (500MHz, DMSO-d6) δ12.60(br.s,1H),7.77(s,1H),7.58(app.d,J=7.3Hz,2H),7.51(app.t,J=7.4Hz,2H),7.46(m,1H).

[0616] 13 C NMR (126MHz, DMSO-d6) δ168.4,167.8,133.5,132.3,130.9,130.5,129.8,124.0.

[0617]

[0618] (Z)-5-((1H-indol-3-yl)methylene)thiazolidin-2,4-dione (JXL075)

[0619] 1 H NMR(500MHz,DMSO-d6)δ12.28(s,1H),12.11(s,1H),8.03(s,1H),7.87(d,J =7.3Hz,1H),7.72(s,1H),7.48(d,J=7.6Hz,1H),7.23(m,1H),7.18(m,1H).

[0620] 13 C NMR (126MHz, DMSO-d6) δ168.2,167.8,136.7,129.1,127.3,125.0,123.5,121.5,118.8,116.7,112.9,110.9.

[0621] Experimental details of JXL022 synthesis

[0622]

[0623] To a solution of 4-pyridinecarboxaldehyde (1 mmol, 107 mg) in ethanol (1 mL), ethyl 2-cyanoacetate (1.3 equivalent, 1.3 mmol, 140 μL) and L-proline (40 mol%, 0.4 mmol, 58 mg) were added. The reaction mixture was stirred at 21 °C for 12 h, and a yellow solid gradually precipitated. After the reaction was complete, ice-cold water (2 mL) was added to the reaction flask. The solid was separated by filtration through a Buchner funnel, washed with water (2 mL × 3), and dried to obtain the desired product, ethyl (E)-2-cyano-3-(pyridin-4-yl)acrylate, which could be used in the next step without further purification.

[0624] To a THF (2 mL) solution of (E)-2-cyano-3-(pyridin-4-yl)acrylate (0.21 mmol, 42.4 mg), 0.5 N LiOH solution (3 equivalents, 0.4 mmol, 0.8 mL) was added. The reaction mixture was stirred at 21 °C for 1 h. After the reaction was complete as indicated by TLC, the THF was evaporated. Concentrated HCl was added dropwise to acidify the reaction mixture until the pH was below 1, precipitating a yellow solid. Ice-cold water (5 mL) was added to the reaction mixture, and the solid was separated by filtration through a Buchner funnel and washed with water (5 mL × 3). After vacuum drying, the solid was washed 5 to 10 times with a solvent mixture of 2 mL (hexane / EtOAc = 5:1) and monitored by TLC until nonpolar impurities disappeared. Finally, the purity of the product was checked by NMR. Yield: 64%, 23.4 mg.

[0625] (E)-2-cyano-3-(pyridin-4-yl)ethyl acrylate (JXL022)

[0626] 1 H NMR (500MHz, CDCl3) δ8.81(d,J=5.2Hz,2H),8.18(s,1H),7.74(d,J=5.2Hz,2H),4.41(q,J=7.1Hz,2H),1.41(t,J=7.1Hz,3H).

[0627] 13 C NMR (126MHz, CDCl3) δ161.2,152.0,151.0,137.9,123.2,114.2,108.2,63.2,14.0.

[0628] The following compounds were synthesized via a route similar to that described for JXL022: JXL030, JXL031, JXL032, JXL033, JXL034, JXL042, JXL43, JXL044, JXL045, JXL046, JXL047, JXL048, and JXL049.

[0629]

[0630] (E)-2-Cyano-3-(2-Fluorophenyl)acrylic acid (JXL030)

[0631] 1 H NMR (500MHz, DMSO-d6) δ8.49(s,1H),8.31(t,J=7.4Hz,1H),7.63(m,1H),7.36(t,J=7.4Hz,1H),7.29(m,1H).

[0632] 13 C NMR(126MHz,DMSO-d6)δ162.9,161.5(d,J c-f =256.2Hz), 145.4(d,J) c-f =7.8Hz), 135.0(d,J c-f =9.2Hz),128.7,124.7,119.8(d,J c-f =10.9Hz), 115.8(d,J) c-f =21.9Hz), 114.9, 105.9.

[0633]

[0634] (E)-2-cyano-3-(4-fluorophenyl)acrylic acid (JXL032)

[0635] 1 H NMR (500MHz, DMSO-d6) δ8.30(s,1H),8.10(m,2H),7.29(m,2H).

[0636] 13 C NMR(126MHz,DMSO-d6)δ165.2(d,J c-f =255.2Hz),163.5,153.1,133.3(d,J c-f =9.3Hz), 128.3, 116.0 (d, J) c-f =22.4Hz), 115.3, 103.2.

[0637]

[0638] (E)-2-cyano-3-(3-(trifluoromethyl)phenyl)acrylic acid (JXL033)

[0639] 1H NMR (500MHz, CD3OD) δ8.24 (s, 1H), 8.17 (m, 2H), 7.80 (d, J = 7.8Hz, 1H), 7.70 (app.t, J = 7.8Hz, 1H).

[0640]

[0641] (E)-2-cyano-3-(4-(trifluoromethyl)phenyl)acrylic acid (JXL034)

[0642] 1 H NMR (500MHz, DMSO-d6) δ8.39 (s, 1H), 8.17 (d, J = 7.7Hz, 2H), 7.84 (d, J = 7.7Hz, 2H).

[0643] 13 C NMR(126MHz,DMSO-d6)δ162.8,152.6,135.4,133.1(q,J c-f =32.9Hz),131.0,125.7,123.7(q,J c-f =272.2Hz), 114.8, 106.7.

[0644]

[0645] (E)-2-cyano-3-(3-fluoro-4-methylphenyl)acrylic acid (JXL042)

[0646] 1 H NMR (500MHz, DMSO-d6) δ8.28 (s, 1H), 7.78 (m, 2H), 7.49 (t, J = 8.0Hz, 1H), 2.30 (s, 3H).

[0647] 13 C NMR(126MHz,DMSO-d6)δ163.6,160.9(d,J c-f =244.6Hz),153.5,133.0,131.6(d,J c-f =7.5Hz), 130.8(d,J c-f =17.6Hz), 127.2, 117.9 (d, J) c-f =23.9Hz), 116.4, 104.5, 15.0.

[0648]

[0649] (E)-2-cyano-3-(3,4-difluorophenyl)acrylic acid (JXL043)

[0650] 1 H NMR (500MHz, DMSO-d6) δ8.32(s,1H),8.09(d,J=8.0Hz,1H),7.94(br.s,1H),7.67(d,J=8.8Hz,1H).

[0651] 13 C NMR(126MHz,DMSO-d6)δ163.3,152.3,152.2(dd,J c-f =255.4,12.6Hz),149.9(dd,J c-f =248.2,12.6Hz),129.8,128.8,120.1(d,J c-f =17.6Hz), 119.1(d,J c-f =17.6Hz), 116.3, 105.8.

[0652]

[0653] (E)-2-cyano-3-(2,4-difluorophenyl)acrylic acid (JXL044)

[0654] 1 H NMR (500MHz, DMSO-d6) δ14.2(br.s,1H),8.27(s,1H),8.24(m,1H),7.52(m,1H),7.35(dt,J=8.6,2.2Hz,1H).

[0655] 13 C NMR(126MHz,DMSO-d6)δ165.5(dd,J c-f =255.8,12.6Hz),163.1,161.9(dd,J c-f =270.5,12.6Hz),145.1,131.1,117.0,115.9,113.5(d,J c-f =22.7Hz), 106.8, 105.6(t,J) c-f =26.5Hz).

[0656]

[0657] (E)-3-(3,5-bis(trifluoromethyl)phenyl)-2-cyanoacrylate (JXL045)

[0658] 1H NMR (500MHz, CD3OD) δ8.59(s,2H),8.48(s,1H),8.19(s,1H).

[0659]

[0660] (E)-3-(2-chloro-3-(trifluoromethyl)phenyl)-2-cyanoacrylate (JXL046)

[0661] 1 H NMR (500MHz, DMSO-d6) δ8.50 (s, 1H), 8.23 ​​(d, J = 7.7Hz, 1H), 8.05 (d, J = 7.4Hz, 1H), 7.75 (app.t, J = 7.9Hz, 1H).

[0662] 13 C NMR (126MHz, DMSO-d6) δ162.6,150.5,134.4,133.5,132.1,131.2,128.9,128.3(q,J c-f =30.2Hz), 123.1(q,J c-f =274.1Hz)115.1,111.1.

[0663]

[0664] (E)-2-cyano-3-(4-fluoro-3-(trifluoromethyl)phenyl)acrylic acid (JXL047)

[0665] 1 H NMR (500MHz, DMSO-d6) δ8.46 (dd, J = 7.1, 1.8Hz, 1H), 8.44 (s, 1H), 8.40 (m, 1H), 7.75 (m, 1H).

[0666] 13 C NMR(126MHz,DMSO-d6)δ163.3,161.1(d,J c-f =262.1Hz), 152.2, 137.6 (d, J) c-f =10.1Hz),130.5,129.3,122.6(q,J c-f =272.8Hz), 118.9(d,J) c-f =21.2Hz), 118.2(qd,J c-f =32.9, 12.6 Hz), 116.2, 106.3.

[0667]

[0668] (E)-2-Cyano-3-Phenylacetic Acid (JXL048)

[0669] 1 H NMR (500MHz, DMSO-d6) δ8.33(s,1H),8.02(m,2H),7.59(m,3H).

[0670] 13 C NMR (126MHz, DMSO-d6) δ163.7,154.9,133.6,132.0,131.1,129.8,116.5,104.3.

[0671]

[0672] (E)-2-cyano-3-(4-hydroxyphenyl)acrylic acid (JXL049)

[0673] 1 H NMR (500MHz, DMSO-d6) δ8.16 (s, 1H), 7.95 (d, J = 8.8Hz, 2H), 6.92 (d, J = 8.8Hz, 2H).

[0674] 13 C NMR (126MHz, DMSO-d6) δ164.2,163.0,153.7,133.6,122.8,117.2,116.3,99.2.

[0675] Experimental details of JXL079 synthesis

[0676]

[0677] A flask containing Mg powder (10 mmol, 240 mg) and a stir bar was sealed, evacuated, and then purged with argon three times. Anhydrous diethyl ether (32 mL) and bis(trifluoromethyl)benzyl bromide (8 mmol, 1.46 mL) were added to the reaction flask. The reaction mixture was stirred and refluxed for 30 minutes, then ground and dried acetic acid powder (5 g) was added to the reaction flask. After 1 h, the reaction was complete as shown by TLC. Excess Mg powder was filtered off, and the solvent was evaporated under vacuum. 1 NHCl (20 mL) was added to the residue, and the precipitate was filtered and dried to provide the desired carboxylic acid.

[0678] The flask containing the stir bar was sealed, evacuated, and then purged with argon three times. Anhydrous dichloromethane (20 mL) and DIBAL (1 M in hexane, 6 mmol, 6 mL) were added to the flask. At -78 °C, crude carboxylic acid (2 mmol, 544 mg) dissolved in dry dichloromethane (10 mL) was added to the reaction flask. After 2 h, the reaction was complete as shown by TLC, and then quenched by adding saturated ammonium chloride (10 mL). The resulting mixture was extracted with dichloromethane (20 mL × 3), the organic phases were combined, and evaporated on a rotary evaporator. The residue was purified by rapid column chromatography (hexane:ethyl acetate = 10:1) to give the desired product 2-(3,5-bis(trifluoromethyl)phenyl)ethane-1-ol (yield: 90%, 464 mg).

[0679] At 0 °C, triethylamine (0.22 mmol, 31 μL) and methanesulfonyl chloride (MsCl, 0.2 mmol, 17 μL) were added to a solution of 2-(3,5-bis(trifluoromethyl)phenyl)ethyl-1-ol (0.2 mmol, 51.6 mg) in dichloromethane (2 mL). After stirring for 1 h, the reaction was complete as shown by TLC. The solvent was removed by passing air through an open flask. The residue was purified by rapid column chromatography (hexane:ethyl acetate = 10:1) to give the desired product, 3,5-bis(trifluoromethyl)phenylethyl methanesulfonate (yield: 82%, 55.1 mg).

[0680] A flask containing NaH (60%, 0.22 mmol, 8.8 mg) and a stir bar was sealed, evacuated, and then re-primed with argon three times. Anhydrous THF (3 mL) and a THF solution of (E)-2-cyano-3-(1H-indol-3-yl)acrylate (0.2 mmol, 53.6 mg) in 2 mL of THF (2 mL) were added to the reaction flask. The reaction mixture was stirred for 30 minutes, and then 3,5-bis(trifluoromethyl)phenylethyl methanesulfonate (0.164 mmol, 55.1 mg) in 2 mL of THF was added. The reaction mixture was stirred for 24 h and quenched with saturated NH4Cl solution. The resulting mixture was extracted with dichloromethane (4 mL × 3), the organic phases were combined, and evaporated on a rotary evaporator. The residue was purified by rapid column chromatography (hexane:ethyl acetate = 10:1) to give the desired product (E)-3-(1-(3,5-bis(trifluoromethyl)phenethyl)-1H-indol-3-yl)-2-cyanoacrylate tert-butyl ester (yield: 68%, 69 mg).

[0681] To a solution of (E)-3-(1-(3,5-bis(trifluoromethyl)phenethyl)-1H-indol-3-yl)-2-cyanoacrylate methyl tert-butyl ester (0.1 mmol, 50.8 mg) in dichloromethane (2 mL), trifluoroacetic acid (3 equivalents, 0.3 mmol, 34 μL) was added. The reaction mixture was stirred at 21 °C for 30 min, and a yellow solid precipitated. After the reaction was complete as indicated by TLC, the reaction solvent was evaporated by passing air through an open flask. The solid was washed 5 to 10 times with a solvent mixture (hexane / EtOAc = 5:1) and monitored by TLC until all nonpolar impurities disappeared. Finally, the purity of the product was checked by NMR. Yield: 87%, 39 mg.

[0682] (E)-3-(1-(3,5-bis(trifluoromethyl)phenethyl)-1H-indol-3-yl)-2-cyanoacrylate (JXL079)

[0683] 1 H NMR(500MHz,CD3OD)δ8.51(s,1H),8.14(s,1H),7.83(d,J=7.3Hz,1H),7.74(s ,1H),7.52(m,3H),7.31(m,2H),4.64(t,J=6.4Hz,2H),3.35(t,J=6.3Hz,2H).

[0684] 13 C NMR (126MHz, CD3OD) δ165.3,145.3,141.1,136.1,133.6,131.3(q,J c-f =32.8Hz),129.3,128.3,123.7,123.3(q,J c-f =272.5Hz),122.3,120.3,118.1,117.6,110.6,109.6,94.1,47.8,35.0.

[0685] Experimental details of JXL080 synthesis

[0686]

[0687] At 0 °C, 2-(3,5-bis(trifluoromethyl)phenyl)acetic acid (0.55 mmol, 150 mg), DMAP (catalytic amount, 6 mg), and DCC (0.5 mmol, 103 mg) were added to a solution of (E)-2-cyano-3-(1H-indol-3-yl)acrylate (0.5 mmol, 112 mg) in dichloromethane (5 mL). The mixture was brought to 21 °C and stirred overnight. The white precipitate was filtered off, and the resulting solution was concentrated under vacuum. The solid was purified by rapid column chromatography (hexane:ethyl acetate = 10:1) to give the desired product (yield: 78%, 192.6 mg).

[0688] (E)-3-(1-(2-(3,5-bis(trifluoromethyl)phenyl)acetyl)-1H-indol-3-yl)-2-cyanoacrylate (JXL080)

[0689] 1 H NMR (500MHz, CDCl3) δ8.90 (s, 1H), 8.52 (s, 1H), 8.49 (d, J = 8.1Hz, 1H), 7.88 (s, 1H), 7.86 (s, 2H) ,7.78(d,J=7.5Hz,1H),7.47(m,2H),4.50(s,2H),4.41(q,J=7.1Hz,2H),1.43(t,J=7.1Hz,3H).

[0690] 13 C NMR (126MHz, CDCl3) δ168.0,162.2,144.2,135.6,134.4,132.3(q,J c-f =33.6Hz),130.1,129.8,128.8,127.8,127.3,125.5,123.1(q,J c-f =273.3Hz),118.3,117.1,117.0,115.9,101.8,62.8,41.9,14.3.

[0691] Experimental details of JXL083 synthesis

[0692]

[0693] To a solution of 1-(3,5-bis(trifluoromethyl)benzyl)-1H-indole-3-carboxaldehyde (10 mmol, 3.71 g) in acetone (60 mL), 2-methyl-2-butene (9 mL), NaH₂PO₄ (3 equivalents, 4.4 g), and NaClO₂ (6.6 mmol, 6 g) in 6 mL of water were added. The reaction mixture was stirred at 21 °C for 24 h. After the reaction was complete as indicated by TLC, the reaction solvent was evaporated on a rotary evaporator. The crude product was dissolved in ethyl acetate (30 mL) and water (30 mL), and extracted with ethyl acetate (30 mL × 3). The organic phases were combined, dried over sodium sulfate, and evaporated on a rotary evaporator. The solid was purified by rapid column chromatography (hexane:ethyl acetate = 2:1) to give the desired product 1-(3,5-bis(trifluoromethyl)benzyl)-1H-indole-3-carboxylic acid (yield: 89%, 3.44 g).

[0694] Seal a 100 mL round-bottom flask equipped with a stir bar, evacuate it, and then refill it with argon three times. The flask contains the carboxylic acid (5 mmol, 1935 mg) from the previous step. Add 50 mL of dichloromethane and oxalyl chloride (25 mmol, 2.1 mL) dropwise to the flask. Stir the reaction mixture at 21 °C for 1.5 h. Evaporate the reaction solvent under vacuum, and use the resulting compound in the next step.

[0695] A 100 mL round-bottom flask equipped with a stir bar was sealed, evacuated, and then re-primed with argon three times. Diisopropylamine (5.5 mmol, 765 μL) and THF (10 mL) were added to the flask, and the mixture was cooled to 78 °C. nBuLi (2.5 M in hexane, 5 mmol, 2 mL) was slowly added to the flask. After stirring the mixture for 30 min, a THF (10 mL) solution of ethyl 2-cyanoacetate (5 mmol, 590 μL) was slowly added to the flask. After stirring the mixture for 1 h, a THF (5 mL) solution of acyl chloride (from the previous step, 5 mmol) was slowly added to the reaction mixture. After 1 h, the reaction was quenched by adding 1 M HCl aqueous solution (10 mL) and extracted with ethyl acetate (10 mL × 3). The organic phases were combined, dried over sodium sulfate, and evaporated using a rotary evaporator. The solid was purified by rapid column chromatography (hexane:ethyl acetate = 10:1) to obtain the desired product (Z)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyano-3-hydroxy ethyl acrylate (JXL083) (yield: 80%, 1.93 g).

[0696] 1H NMR (500MHz, CDCl3) δ14.61 (s, 1H), 8.67 (s, 1H), 8.31 (dd, J = 7.0 1.3Hz,1H),7.84(s,1H),7.57(s,2H),7.33(m,2H),7.21(d,J=7.4Hz,1H),5.53(s,2H),4.40(q,J=7.1Hz,2H),1.41(t,J=7.1Hz,3H).

[0697] 13 C NMR (126MHz, CDCl3) δ179.1,172.2,138.1,136.0,135.7,132.7(q,J c-f =33.6Hz),126.9,124.5,123.7,123.6,122.6,122.9(q,J c-f =273.4Hz),121.8,118.3,110.1,109.6,73.1,62.3,50.4,14.3.

[0698] Experimental details of JXL084 synthesis

[0699]

[0700] To a solution of JXL083 (0.5 mmol, 240 mg) in dichloromethane (10 mL), pyridine (0.5 mmol, 40 μL) and acetyl chloride (1.0 mmol, 84 μL) were added. The reaction mixture was stirred for 1 h, and TLC indicated that the reaction was complete. The reaction solvent was evaporated by passing air through an open flask. The residue was purified by rapid column chromatography (hexane:ethyl acetate = 10:1) to give the desired product (Z)-3-acetoxy-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyanoacrylate (JXL084) (yield: 86%, 225 mg).

[0701] 1 H NMR(500MHz, CDCl3)δ8.56(s,1H),7.90(m,1H),7.85(s,1H),7.61(s,2H),7.32(m,2H) ,7.24(m,1H),5.51(s,2H),4.29(q,J=7.1Hz,2H),2.48(s,3H),1.36(t,J=7.1Hz,3H).

[0702] 13 C NMR (126MHz, CDCl3) δ166.9,161.4,137.7,136.2,135.5,132.8(q,Jc-f =33.9Hz),127.0,126.7,124.6,123.6,122.8(q,J c-f =273.4Hz),122.7,121.8,117.7,110.7,109.8,89.3,61.8,50.5,29.7,21.3,14.2.

[0703] Experimental details of JXL085 synthesis

[0704]

[0705] Triethylamine (1.0 mmol, 139.5 μL) and phosphoryl chloride (0.55 mmol, 520 μL) were added to a solution of JXL083 (0.5 mmol, 240 mg) in dichloromethane (10 mL). The reaction mixture was stirred under reflux for 1 h, and TLC indicated that the reaction was complete. The reaction solvent was evaporated by passing air through an open flask. The residue was purified by rapid column chromatography (hexane:ethyl acetate = 10:1) to give the desired product (Z)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-3-chloro-2-cyanoacrylate (JXL085) (yield: 84%, 210 mg).

[0706] 1 H NMR(500MHz, CDCl3)δ7.91(s,1H),7.85(s,1H),7.77(m,1H),7.65(s,2H),7.63 (s,1H),7.30(m,2H),5.48(s,2H),4.18(q,J=7.1Hz,2H),1.18(t,J=7.1Hz,3H).

[0707] 13 C NMR (126MHz, CDCl3) δ161.0,155.6,138.1,136.2,135.4,133.4,132.7(q,J c-f =33.7Hz),127.1,126.6,124.5,124.3,122.8(q,J c-f =273.4Hz),121.5,115.8,122.1,110.4,102.5,62.4,50.0,13.9.

[0708] Experimental details of JXL086 synthesis

[0709]

[0710] (E)-(2-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-1-cyanovinyl)phosphonate diethyl ester (JXL086)

[0711] To a solution of 1-(3,5-bis(trifluoromethyl)benzyl)-1H-indole-3-carboxaldehyde (1 mmol, 371 mg) in ethanol (3 mL), diethyl cyanomethyl phosphate (1.3 equivalent, 1.3 mmol, 204 μL) and L-proline (40 mol%, 0.4 mmol, 58 mg) were added. The reaction mixture was stirred at 50 °C for 24 h. After the reaction was indicated by TLC, the reaction solvent was evaporated by passing air through an open flask. The solid was purified by rapid column chromatography (hexane:ethyl acetate = 2:1) to give the desired product JXL086 (yield: 90%, 477 mg).

[0712] 1 H NMR (500MHz, CDCl3) δ8.55(s,1H),8.33(d,J=19.7Hz,1H),7.86(d,J=7.9Hz,1H),7.83(s,1 H),7.58(s,2H),7.31(m,2H),7.22(m,1H),5.54(s,2H),4.21(m,4H),1.39(t,J=7.0Hz,6H).

[0713] 13 C NMR (126MHz, CDCl3) δ149.8,138.3,135.8,132.7(q,J c-f =33.7Hz),132.6,128.2,126.8,124.5,123.0,122.9(q,J c-f =273.4Hz),122.5,119.0,117.8(d,J c-p =11.3Hz), 112.2(d,J c-p =18.9Hz), 110.3, 91.6(d,J) c-p =207.9Hz), 63.2, 50.4, 16.3.

[0714] The synthetic route of JXL095 is similar to that of JXL086.

[0715]

[0716] (E)-(2-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-1-cyanovinyl)phosphonate diethyl ester (JXL095)

[0717] 1H NMR (500MHz, CDCl3) δ8.58 (s, 1H), 8.46 (dd, J = 4.7, 1.4Hz, 1H), 8.24 (d, J = 19.5Hz 1H),8.20(dd,J=8.0,1.4Hz,1H),7.82(s,1H),7.77(s,2H),7.32(dd,J=8.0,4.7Hz,1H),5.67(s,2H),4.22(m,4H),1.40(t,J=7.1Hz,6H).

[0718] 13 C NMR(126MHz,CDCl3)δ149.2(d,J c-p =8.2Hz),147.4,145.6,138.6,132.4(q,J c-f =33.7Hz),132.0,127.9,127.8,123.0(q,J c-f =273.4Hz),122.4,119.9,118.9,117.4(d,J c-p =11.3Hz), 110.5(d,J c-p =19.5Hz), 92.8(d,J c-p =205.1Hz), 63.4, 48.3, 16.3.

[0719] Experimental details of JXL096 synthesis

[0720]

[0721] (E)-(2-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-1-cyanovinyl)phosphonic acid (JXL096)

[0722] A solution of JXL086 (30 mg, 0.057 mmol) in dichloromethane (2 mL) was cooled to 0 °C, and trimethylbromosilane (40 μL, 0.3 mmol) was added dropwise under argon. The mixture was heated to 21 °C and stirred for 12 h. The solvent was evaporated under vacuum, and the resulting residue was then dissolved in methanol (2 mL). The mixture was stirred at 21 °C for 2 h. All volatiles were evaporated under vacuum to give JXL096 phosphoric acid (yield: 92%, 25 mg).

[0723] 1H NMR (500MHz, CD3OD) δ8.58(s,1H),8.25(d,J=19.6Hz,1H),7.90(s,1H),7.87(m,1H),7.76(s,2H),7.45(m,1H),7.31(m,2H),5.75(s,2H).

[0724] 13 C NMR (126MHz, CD3OD) δ 146.7 (J c-p =7.2Hz),140.2,136.1,132.1,131.9(q,J c-f =33.7Hz),128.0,127.2,123.8,122.3,123.2(q,J c-f =273.4Hz),121.4,118.2,117.4(d,J c-p =11.3Hz), 111.5(J) c-p =18.4Hz), 110.5, 94.6 (d, J) c-p =201.2Hz), 49.1.

[0725] Experimental details of JXL092 synthesis

[0726]

[0727] To a 3 mL ethanol solution (1 mmol, 429 mg) of methyl 1-(3,5-bis(trifluoromethyl)benzyl)-3-formyl-1H-indole-4-carboxylic acid, tert-butyl 2-cyanoacetate (1.3 equivalent, 1.3 mmol, 183 μL) and L-proline (40 mol%, 0.4 mmol, 58 mg) were added. The reaction mixture was stirred at 21 °C for 12 h, and a yellow solid gradually precipitated. After the reaction was complete, ice-cold water (2 mL) was added to the reaction mixture. The solid was separated by filtration through a Buchner funnel, washed with water (2 mL × 3), and dried to obtain the desired product. Yield: 95%, 524 mg.

[0728] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-4-(methoxycarbonyl)-1H-indol-3-yl)-2-cyanoacrylate (JXL092)

[0729] To a solution of (E)-1-(3,5-bis(trifluoromethyl)benzyl)-3-(3-(tert-butoxy)-2-cyano-3-oxoprop-1-en-1-yl)-1H-indole-4-carboxylic acid methyl ester (0.5 mmol, 276 mg) in dichloromethane (2 mL), trifluoroacetic acid (3 equivalents, 1.5 mmol, 0.2 mL) was added. The reaction mixture was stirred at 21 °C for 30 min, and a yellow solid precipitated. After the reaction was complete as indicated by TLC, the reaction solvent was evaporated by passing air through an open flask. The solid was washed 5 to 10 times with a solvent mixture (hexane / EtOAc = 5:1) and monitored by TLC until all nonpolar impurities disappeared. Finally, the purity of the product was checked by NMR. Yield: 90%, 223 mg.

[0730] 1 H NMR (500MHz, CDCl3) δ9.09 (s, 1H), 8.58 (s, 1H), 7.75 (d, J = 7.5Hz, 1H), 7.68 (s, 1H), 7. 47(s,2H),7.34(d,J=8.2Hz,1H),7.20(app.t,J=7.9Hz,1H),5.51(s,2H),3.87(s,3H).

[0731] 13 C NMR (126MHz, CDCl3) δ167.8,165.1,149.5,138.2,137.2,134.8,132.3(q,J c-f =33.7Hz),126.8,126.2,125.5,124.8,123.3,122.8(q,J c-f =273.4Hz),122.3 118.2,114.7,110.9,97.2,52.4,50.2.

[0732] Experimental details of JXL094 synthesis

[0733]

[0734] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyanoacrylamide (JXL094)

[0735] A mixture of (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyanoacrylate JXL001 (0.1 mmol, 43.8 mg) and thionyl chloride (0.5 mL) was refluxed for 1 h. After vacuum concentration, the resulting acyl chloride was dissolved in 1 mL of benzene, and 30% ammonia (1 mL) was added. The reaction mixture was stirred at 21 °C for 24 h. After the reaction was indicated by TLC to be complete, the reaction solvent was evaporated under vacuum. The solid was purified by rapid column chromatography (hexane:ethyl acetate = 2:1) to give the desired product JXL094 (yield: 85%, 37 mg).

[0736] 1 H NMR (500MHz, CDCl3) δ8.70 (s, 1H), 8.47 (s, 1H), 7.92 (d, J = 7.1Hz, 1H), 7.85 (s, 1H), 7.58 (s, 2H), 7.34 (m, 2H), 7.23 (m, 1H), 5.55 (s, 2H).

[0737] 13 C NMR (126MHz, CDCl3) δ163.0,144.8,138.1,136.0,132.7(q,J c-f =33.7Hz),132.5,128.6,126.8,126.1,124.6,123.1,122.9(q,J c-f =273.4Hz),122.6,119.3,111.3,110.3,96.1,50.4.

[0738] Other exemplary compounds of the present invention can be prepared by methods similar to those described above.

[0739] Example 2: Treatment of epithelial cells with exemplary compounds

[0740] To determine whether these compounds promote cellular lactate production, we treated cultured epithelial cells with the compounds and measured lactate levels in the culture medium using the Nova Biomedical BioProfile Basic Analyzer. In short, cultured epithelial cells were treated with DMSO, UK-5099 (also known as JXL001), or certain exemplary compounds disclosed herein for 24–30 hours, and then the lactate levels in the culture medium were measured and normalized to cell number and experimental duration to obtain the cellular lactate production rate (nmol lactate, million cells, hours).

[0741] The rate of lactate production in the treated cells is shown in Figure 8 , Figure 9 and Figure 12 In, as anticipated based on this disclosure, most of the new compounds showed increased lactate production because they are UK-5099 analogs. Furthermore, the total cell count after treatment with the UK-5099 analogs is shown in... Figure 13 In China, most compounds are tolerated by cells. For example... Figure 10 As shown, individual measurements were performed to calculate the EC50 of some compounds.

[0742] Example 3: Exemplary compounds in vivo test

[0743] To determine the efficacy of the compound on the hair cycle, mice were shaved on day 50 after birth and treated topically every other day with the compound disclosed herein suspended in a shampoo solution for two weeks, followed by photography. Figure 11 As shown, in in vitro assays, all analogues that promote lactic acid production were also shown to stimulate hair growth over a 2-week period.

[0744] By incorporating references

[0745] All publications and patents mentioned herein are hereby incorporated by reference in their entirety, as if each individual publication or patent were specifically and individually indicated to be incorporated by reference. In case of any conflict, this application (including any definitions herein) shall prevail.

[0746] equivalent

[0747] While specific embodiments of the invention have been discussed, the foregoing description is illustrative rather than restrictive. Many variations of the invention will become apparent to those skilled in the art upon review of this description and the following claims. The full scope of the invention should be determined by reference to the full scope of the claims and their equivalents, the description, and such variations.

Claims

1. A cosmetic method of promoting hair growth comprising administering to a subject a compound, wherein the compound is or a pharmaceutically acceptable salt thereof.

2. The cosmetic method of claim 1, wherein the compound is 3. The cosmetic method of claim 1, wherein the compound is a pharmaceutically acceptable salt of .

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