Compositions and methods for modulating hair growth

Abstract

To provide compounds that are capable of inhibiting the mitochondrial pyruvate carrier and promoting hair growth, pharmaceutical compositions, and methods of promoting hair growth.SOLUTION: Provided is a compound represented by formula I or II (where, each A is CH, CR4, or N; Y is carboxyl, ester, amide, or -P(=O)(OR)2; Z is CH, CR4, or N; R2 is CN or carboxyl; R3 is H, aryl, aralkyl, or aralkylacyl; R7 is H or alkyl; and n is 0-4).SELECTED DRAWING: Figure 7A

Description

Technical Field

[0001] Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 62 / 527,775, filed Jun. 30, 2017, and U.S. Provisional Patent Application No. 62 / 654,095, filed Apr. 6, 2018. The entire content of each of these applications is hereby incorporated by reference herein.

Background Art

[0002] Hair follicle stem cells (HFSCs) undergo a continuous repetition of a quiescent state (telogen) interrupted by short-term proliferation correlated with the onset of the hair cycle (telogen-anagen transition). The proliferation or activation of HFSCs is well known to be a prerequisite for the progression of the hair cycle. Despite advances in treatment options, alopecia and hair loss remain conditions that cannot be successfully treated in all individuals. Some of the existing treatments are inconvenient for the user, and others require surgical procedures or other invasive procedures. Additional therapies are needed.

Summary of the Invention

[0003] In certain aspects, the disclosure provides a compound of Formula I or II,

[0004]

Chemical Formula

[0005]

Chemical Formula

[0006] In certain embodiments, the disclosure is a compound of formula III,

[0007]

Chemical formula

[0008]

Chemical formula

[0009] In certain embodiments, the disclosure is a compound of formula V, VI, or VII,

[0010]

Chemical formula

[0011] In certain embodiments, the disclosure provides a compound of Formula Va, VIa, or VIIa,

[0012]

Chemical formula

[0013] In certain aspects, the disclosure provides a pharmaceutical composition comprising a compound of the disclosure and a pharmaceutically acceptable excipient.

[0014] In certain aspects, the disclosure provides a method of enhancing intracellular lactic acid production, the method comprising contacting a cell with a compound or composition of the disclosure.

[0015] In certain embodiments, the present disclosure provides a method of inhibiting mitochondrial pyruvate oxidation in a cell, the method comprising contacting the cell with a mitochondrial pyruvate oxidation (MPO) inhibitor such as a compound of the present disclosure. In certain embodiments, the MPO inhibitor is a mitochondrial pyruvate carrier (MPC) inhibitor. In certain embodiments, inhibition of mitochondrial pyruvate oxidation in the cell enhances intracellular lactate production and / or enhances the activity of intracellular LDH, and has the effect of promoting hair growth, as described herein.

[0016] In certain embodiments, the present disclosure provides a method of enhancing intracellular lactate production, the method comprising contacting the cell with an MPO inhibitor such as a compound of the present disclosure. In certain embodiments, the MPO inhibitor is a mitochondrial pyruvate carrier (MPC) inhibitor.

[0017] In certain embodiments, the present disclosure provides a method of enhancing the activity of intracellular LDH, the method comprising contacting the cell with an MPO inhibitor such as a compound of the present disclosure. In certain embodiments, the MPO inhibitor is a mitochondrial pyruvate carrier (MPC) inhibitor.

[0018] In certain embodiments, the present disclosure provides a method of enhancing the activity of intracellular lactate dehydrogenase (LDH), the method comprising contacting the cell with an MPO inhibitor such as a compound of the present disclosure. In certain embodiments, the MPO inhibitor is a mitochondrial pyruvate carrier (MPC) inhibitor. In certain embodiments, the present disclosure provides a method of promoting hair growth, or treating a hair disorder or condition such as alopecia or hair loss, the method comprising administering to a patient a compound or composition disclosed herein.

[0019] In certain embodiments, the present disclosure provides a method for promoting hair growth or treating hair disorders or conditions such as alopecia or hair loss, the method comprising administering to a patient an MPO inhibitor, such as a compound of the present disclosure, (e.g., topically, as in the case of a pharmaceutical composition formulated for topical administration) to the patient. In certain embodiments, the present disclosure provides a method for promoting hair growth or treating hair disorders or conditions such as alopecia or hair loss, the method comprising administering to a patient an MPC inhibitor, such as a compound of the present disclosure, (e.g., topically, as in the case of a pharmaceutical composition formulated for topical administration) to the patient. In certain embodiments, inhibition of mitochondrial pyruvate oxidation or mitochondrial pyruvate carrier within the cell enhances intracellular lactate production and / or enhances the activity of LDH and has the effect of promoting hair growth, as described herein.

Brief Description of the Drawings

[0020]

Figure 1A

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DETAILED DESCRIPTION OF THE INVENTION

[0021] In certain aspects, the disclosure provides a compound of Formula I or II, wherein

[0022]

CHEMICAL

[0023]

CHEMICAL

[0024] In certain embodiments, the compound is a compound of Formula I. In certain embodiments, the compound is a compound of Formula II.

[0025] In certain embodiments of Formula I or II, Z is CH or N.

[0026] In certain embodiments, the present disclosure is a compound of Formula III or IV,

[0027]

Chemical formula

[0028]

Chemical formula

[0029] In certain embodiments, the compound is a compound of Formula III.

[0030] In certain embodiments of Formulas I, II, and III, Y is, Y is,

[0031]

Chem.

[0032] In certain embodiments of Formula I, II, or III, R 11 is alkyl (e.g., methyl).

[0033] In certain embodiments, the present disclosure is a compound of Formula V, VI, or VII,

[0034]

Chem.

[0035] In certain embodiments, the compound is a compound of formula V. In certain embodiments, the compound is a compound of formula VI. In certain embodiments, the compound is a compound of formula VII.

[0036] In certain embodiments of formulae I, II, III, V, VI, or VII, at least one A is N, preferably no more than two occurrences of A are N. In certain preferred embodiments, exactly one A is N, preferably A, is NR 3 bonded to the same carbon as.

[0037] In certain embodiments, the present disclosure is a compound of formula Va, VIa, or VIIa, wherein

[0038] [Chemical formula] In the formula, X is NR 6 or O, and R 1 is H or lower alkyl, and R 2 is CN or carboxyl, or R 1 and R 2 together with the atoms separating them complete a heterocyclic ring, R 3 is H, phenyl, or benzyl, and is optionally substituted by one or more R 5 where each R 5 is independently selected from alkyl, alkoxy, or halo, each instance of R 4 is independently alkyl, carboxyl, halo, hydroxy, or CN, R 6 is selected from H, alkyl, or cycloalkyl, a compound, and pharmaceutically acceptable salts thereof are provided.

[0039] In certain embodiments, the compound is a compound of formula Va. In certain embodiments, the compound is a compound of formula VIa. In certain embodiments, the compound is a compound of formula VIIa.

[0040] In certain embodiments of formula V, VI, VII, Va, VIa, or VIIa, X is NH. In certain embodiments, X is O.

[0041] In certain embodiments of formula V, VI, VII, Va, VIa, or VIIa, R 1 is H. In certain embodiments, R 1 is lower alkyl. In certain embodiments, R 1 and R 6 together with the atoms separating them complete a heterocyclic ring (e.g., morpholinyl).

[0042] In certain embodiments of Formulas V, VI, VII, Va, VIa, or VIIa, R 6 is hydrogen.

[0043] In certain embodiments of Formulas V, VI, VII, Va, VIa, or VIIa, R 2 is CN. In certain embodiments, R 2 is carboxyl. In certain embodiments, R 1 and R 2 together with the atom separating them complete a heterocyclyl selected from thiazolidine-2,4-dione-5-ylidene or 2-iminothiazolidin-4-one-5-ylidene.

[0044] In certain embodiments of Formulas I, II, III, V, VI, VII, Va, VIa, or VIIa, R 3 is H. In certain embodiments, R 3 is phenyl. In certain embodiments, R 3 is phenyl and is substituted by one or more R 5 . In certain embodiments, R 3 is substituted by one R 5 , where R 5 is alkoxy. In certain embodiments, R 3 is aralkyl (e.g., benzyl or phenethyl). In certain embodiments, R 3 is aralkyl acyl (e.g., phenylacetyl). In certain embodiments, R 3 is benzyl. In certain embodiments, R 3 is benzyl and is substituted by one or more R 5 . In certain embodiments, R 3 is aralkyl (e.g., benzyl or phenethyl) and is substituted by one or more R 5 (preferably on the phenyl ring). In certain embodiments, R 3 is aralkyl acyl (e.g., phenylacetyl) and is substituted by one or more R 5is replaced by. In certain embodiments, R 3 is substituted by one or two R 5 each R 5 is independently selected from fluoroalkyl or fluoro. In certain embodiments, R 3 is substituted by two R 5 each R 5 is trifluoromethyl.

[0045] In certain embodiments of Formula Va, VIa, or VIIa, n is 0.

[0046] In certain preferred embodiments, the present disclosure provides a compound of Formula Vb.

[0047]

Chemical formula

[0048] In certain embodiments, the present disclosure provides a compound of Formula Vc.

[0049]

Chemical formula

[0050] In certain embodiments of Formula Va, VIa, VIIa, or Vb, n is 1.

[0051] In certain preferred embodiments, the present disclosure provides a compound of Formula Vd.

[0052]

Chemical formula

[0053] In certain embodiments, the present disclosure provides a compound of Formula Ve.

[0054]

Chemical formula

[0055] In certain embodiments of Formula Vb, Vd, or Ve, R 4 is selected from halo or haloalkyl. In certain preferred embodiments, R 4 is halo (e.g., chloro or bromo). In other preferred embodiments, R 4 is carboxyl or ester.

[0056] In certain embodiments of Formula VI or VIa, n is 0. In certain embodiments, n is 2 and R 4 is selected from halo or haloalkyl.

[0057] In certain embodiments of Formula I, II, III, V, VI, VII, Va, Vb, Vc, Vd, Ve, VIa, or VIIa, R 7 is hydrogen, hydroxyl, halo (e.g., chloro), or acyloxy (e.g., acetyloxy). In certain embodiments, R 7 is hydroxyl, halo (e.g., chloro), or acyloxy (e.g., acetyloxy).

[0058] In certain embodiments of Formula I, II, III, V, VI, VII, Va, Vb, Vc, Vd, Ve, VIa, or VIIa, the compound is not JXL001.

[0059] In certain aspects, the compounds of the present disclosure are the compounds of Table 1. Table 1: Exemplary Compounds of the Invention

[0060]

Chemical Formula

[0061] In certain aspects, the present disclosure provides a pharmaceutical composition comprising a compound of the present disclosure and a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition is formulated for topical administration.

[0062] In certain aspects, the present disclosure provides a method of enhancing intracellular lactate production, the method comprising contacting a cell with a compound or composition of the present disclosure. In certain embodiments, the cell is a hair follicle stem cell.

[0063] In certain aspects, the present disclosure provides a method of promoting hair growth or treating a hair disorder or condition such as alopecia or hair loss, the method comprising administering to a patient a compound or composition disclosed herein.

[0064] In certain aspects, the present disclosure provides a method of enhancing intracellular lactate production, the method comprising contacting a cell with a mitochondrial pyruvate carrier (MPC) inhibitor. In certain embodiments, the MPC is MPC1.

[0065] In certain aspects, the present disclosure provides a method of promoting hair growth or treating a hair disorder or condition such as alopecia or hair loss, the method comprising administering to a patient (e.g., topically, as in the case of a pharmaceutical composition formulated for topical administration) a mitochondrial pyruvate carrier (MPC) inhibitor. In certain embodiments, the MPC is MPC1.

[0066] In certain aspects, the present disclosure provides a method of inhibiting mitochondrial pyruvate oxidation in a cell, the method comprising contacting the cell with a mitochondrial pyruvate oxidase (MPO) inhibitor.

[0067] In certain aspects, the present disclosure provides a method for promoting hair growth or treating hair disorders or conditions such as alopecia or hair loss, the method comprising administering to a patient a mitochondrial pyruvate carrier (MPC) or mitochondrial pyruvate oxidation (MPO) inhibitor (e.g., topically, as in the case of a pharmaceutical composition formulated for topical administration).

[0068] In certain embodiments, the MPC or MPO inhibitor is a compound of the present disclosure.

[0069] Discussion Numerous studies have revealed gene expression characteristics specific to hair follicle stem cells (HFSCs) relative to other hair follicle or interfollicular epidermal cells. Many of these characteristics are controlled by transcription factors that are later shown to play important roles in HFSC homeostasis.

[0070] Lactate dehydrogenase is most commonly encoded by the Ldha and Ldhb genes in mammals, and these protein products form homotetramers or heterotetramers that catalyze the NADH-dependent reduction of pyruvate to lactate and the NAD + -dependent oxidation of lactate to pyruvate. By immunostaining, Ldha has been found to be enriched within HFSCs in the in situ quiescent (telogen) state (Figure 1A), and performance of immunohistochemical staining (IHC) with an antibody that recognizes both Ldha and Ldhb indicated that only Ldha localizes within the HFSC niche (Figure 2A).

[0071] IHC analysis also showed that Ldha expression was enriched within HFSCs (Sox9+) at three stages of the hair cycle (Figure 1A). As a result, immunoblots of lysates from sorted cells showed expression of Ldha at enhanced levels within basal HFSCs (α6high CD34+) and supra-basal (α6low CD34+) HFSC populations compared to total epidermis (Figure 1B) (an overview of the sorting strategy is described in Figure 2B).

[0072] To determine whether the Ldha expression pattern correlates with the activity of the Ldh enzyme, an in situ Ldh activity assay was performed using a colorimetric-based enzyme assay to evaluate the Ldh activity capacity in situ. The Ldh activity assay is typically performed on protein lysates or aliquots using a plate reader, but was adapted to function on frozen tissue sections in situ. It should be noted that for both in situ and in vitro Ldh activity assays, since an excess amount of substrate (lactic acid) is used, the results from these assays reflect the capacity of Ldh activity rather than the activity at steady state.

[0073] By applying this assay to skin samples, it was demonstrated that, consistent with the expression pattern of Ldha, the capacity of Ldh activity was significantly higher within HFSCs (Figure 1C). Furthermore, Ldh activity was enriched within HFSCs over the hair cycle (Figure 1C). As a control, the activity resulting from assays performed without the enzyme substrate (lactic acid) or on acid-treated tissue was zero (Figure 2C). To further validate these results, we sorted epidermal populations, generated cell lysates on the sorted cells, and performed a similar colorimetric-based enzyme assay on the lysates of the sorted cells, which also showed an increase in Ldh activity within HFSCs (Figure 1D). To better characterize the metabolism of HFSCs, we performed metabolome analysis on sorted populations derived from mouse skin by liquid chromatography-mass spectrometry (LC-MS) (Figure 1E). Several glycolytic metabolites, including glucose / fructose-6-phosphate, fructose-diphosphate, dihydroxyacetone phosphate, 3-phosphoglyceric acid, and lactic acid, were always higher within HFSCs compared to the whole epidermis over three independent experiments (isolated from different mice on different days). Conversely, most TCA cycle metabolites did not consistently differ between the epidermis and HFSCs (Figure 1E). Collectively, these results suggest that while all cells within the epidermis extensively use the TCA cycle to generate energy, HFSCs also have increased Ldha expression, Ldh activity, and glycolytic metabolism.

[0074] Therefore, measuring metabolism over the hair cycle will capture any dynamic changes occurring within HFSCs that correlate with activation or quiescence. Analysis of RNA-seq data from HFSCs isolated either during quiescence or during the quiescence-to-growth transition demonstrated not only that Ldha is the dominant Ldh isoform expressed within HFSCs (Figure 3), but also that it is induced during the quiescence-to-growth transition (Figures 3A and 3B) (NIH GEO GSE67404 and GSE51635). To confirm that the cells analyzed by RNA-seq were indeed in either quiescence or the quiescence-to-growth transition, we evaluated key markers of this transition, including the Shh and Wnt pathways (Gli1, 2, 3; Lef1, Axin1, Axin2, Ccnd1), as well as proliferation markers (Ki-67, Pcna, and Sox4) (Figure 4A).

[0075] In vitro Ldh activity assays on lysates from sorted HFSCs revealed a moderate induction of Ldh activity correlating with the quiescence-to-growth transition (Figure 3D). We determined HFSC activation by validating hair cycle stage by Ki-67 immunostaining (Figure 4B). In addition, measurements of steady-state metabolites extracted from sorted HFSCs showed that lactate increased within HFSCs as they transitioned from quiescence to the quiescence-to-growth transition and then decreased again during growth as the HFSCs returned to the quiescent state (Figure 3E).

[0076] To determine whether induction of lactate production can affect HFSC activation or the hair cycle, we crossed K15CrePR animals to animals in which loxP was introduced for mitochondrial pyruvate carrier 1 (Mpc1) (K15CrePR;Mpc1 fl / fl) Mpc1, as a heterodimer with Mpc2, forms the mitochondrial pyruvate carrier MPC, a transporter on the inner mitochondrial membrane required for pyruvate entry into the mitochondria. Loss of Mpc1 function has been shown to drive lactate production through enhanced conversion of pyruvate to lactate by Ldh. Furthermore, inhibition of MPC results in a decrease in mitochondrial pyruvate oxidation (MPO) to acetyl coenzyme A (Figure 20).

[0077] In animals with Mpc1 deletion in HFSCs, we observed an acceleration in the intensity of abdominal and dorsal hair cycles, along with all typical features of the quiescence-to-growth transition (Figure 5A) (n = 12 littermate pairs). K15CrePR;Mpc1 animals treated with mifepristone fl / fl did not show any signs of dorsal anagen until day 70. Western blot of sorted HFSCs verified the loss of Mpc1 protein (Figure 5B). Importantly, purified HFSCs lacking Mpc1 showed induction of the intensity of Ldh activity (Figure 5C). Quantification of the dorsal hair cycles across three pairs of littermates showed induction of the intensity of anagen in the dorsal skin lacking Mpc1 (Figure 5D, right), and histology showed a normal appearance with typical subcutaneous tissue expansion due to induction of anagen (Figure 5D). Immunostaining demonstrated induction of various markers of hair cycle activation, such as Ki-67 and pS6, while Sox9 expression was unaffected, in Mpc1-deficient HFSCs (Figure 5E). Long-term deletion of Mpc1 did not result in abnormal hair follicle or HFSC depletion, as judged by pathology and Sox9 staining (Figure 6A). Furthermore, deletion of Mpc1 by Lgr5CreER showed a phenotype very similar to that by K15CrePR (Figure 5F and 5G), verifying the fact that deletion of this protein in HFSCs leads to their activation (n = 12 pairs of littermates). Finally, immunofluorescence of the Ires-GFP of the Lgr5CreER transgene together with Ki-67, and K15CrePR;Mpc1 fl / flLineage tracing with the lsl-Tomato mouse also demonstrated that HFSCs actually proliferate after induction of Mpc1 deletion by tamoxifen and mifepristone (Figure 6B).

[0078] UK-5099 (also referred to herein as JXL001) is an established pharmacological inhibitor of the mitochondrial pyruvate carrier and is known to promote lactate production as a result in various settings. UK-5099 has the following structure.

[0079]

Chemical formula

[0080] Local treatment of animals in telogen (day 50) with UK-5099 resulted in a robust acceleration of the hair cycle and a slight overgrowth of the interfollicular epidermis (Figure 7A). Quantification of the hair cycle over at least 6 pairs of animals (vehicle vs. UK-5099) showed an acceleration of the hair cycle intensity in about 6 - 9 days (Figure 7B). Similar to genetic deletion of Mpc1, pharmacological blockade of the mitochondrial pyruvate carrier with UK-5099 for 48 hours during telogen promoted an increase in Ldh activity within HFSCs and in the interfollicular epidermis, consistent with an increase in the ability to produce lactate (Figure 7C). Finally, metabolome analysis demonstrated that topical application of UK-5099 increased the total levels of lactate within sorted HFSCs (Figure 7D).

[0081] Compounds were synthesized that can locally promote an increase in lactate levels and thus drive the hair cycle.

[0082] This compound was generally prepared by reacting the corresponding aldehyde (e.g., 1-phenylindole-3-carboxaldehyde in the case of JXL001) with ethyl cyanoacetate in the presence of a 40% aqueous solution of L-proline to exclusively yield the E-isomer of ethyl 2-cyano-3-(1-phenylindol-3-yl)propenoic acid (e.g., JXL004). Hydrolysis of the ester with mild lithium hydroxide gave the E-isomer of the acid (e.g., JXL001). All of the other compounds were prepared by similar methods using specific aldehydes. Two heterocyclic compounds, JXL023 and JXL024, were prepared by condensing 1-phenyl-indole-3-carboxaldehyde with thiazolidine-2,4-dione and 2-iminothiazolidin-4-one. All of the structures of the compounds were determined using conventional organic chemistry methods, particularly high-field proton, carbon, and fluorine NMR spectra. Specifically, 3 J C-H Bond measurements demonstrated that all of the compounds had E-stereochemistry for the important carbon-carbon double bonds.

[0083] To determine whether these compounds were able to promote cellular lactate production, we treated cultured epithelial cells with the compounds and measured the lactate levels in the culture medium using a Nova Biomedical BioProfile Basic Analyzer. Briefly, cultured epithelial cells were treated with DMSO, UK-5099 (also called JXL001), or some of the exemplary compounds disclosed herein for 24 - 30 hours, the lactate levels in the medium were measured, and normalized against the cell number and duration of the experiment to obtain the cellular lactate production rate (nmol of lactate, per million cells, per hour). The results are shown in Figures 8 and 9.

[0084] The lactate production rates of the treated cells are shown in Figure 8. As expected, most of the novel compounds assayed increased lactate production since they are UK-5099 analogs. A separate assay was performed to calculate the EC 50 of some of the compounds shown in Figure 10.

[0085] To determine the effectiveness of the compounds against the hair cycle, mice were shaved on the 50th day after birth and topically treated every other day for 3 weeks with the compounds disclosed herein (suspended in lotion). As seen in Figure 11, all analogs that showed the ability to promote lactic acid production in the in vitro assay were also able to stimulate hair growth over a 2-week period.

[0086] Pharmaceutical composition Using the compositions and methods of the present invention, individuals in need of treatment can be treated. In certain embodiments, the individual is a mammal or non-human mammal, such as a human. 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 well known in the art and include, for example, aqueous solutions such as water or physiological buffered saline, or other solvents or vehicles such as glycols, glycerol, oils (such as olive oil), or injectable organic esters. In a preferred embodiment, when such a pharmaceutical composition is for human administration, particularly for invasive routes of administration (i.e., routes such as injection or implantation that avoid transport or diffusion through an epithelial barrier), the aqueous solution is free of pyrogens or substantially free of pyrogens. Excipients can be selected, for example, to provide for delayed release of the drug or to selectively target one or more cells, tissues, or organs. The pharmaceutical composition can be in unit dosage forms such as tablets, capsules (including sprinkle capsules and gelatin capsules), granules, lyophilized products for reconstitution, powders, solutions, syrups, suppositories, or injection solutions. The composition can also be present in a transdermal delivery system, such as a skin patch. The composition can also be present in a solution suitable for topical administration, such as a lotion, cream, or ointment.

[0087] A pharmaceutically acceptable carrier may contain a physiologically acceptable agent that acts, for example, to stabilize a compound such as a compound of the present invention, increase its solubility, or increase its absorption. 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 selection of a pharmaceutically acceptable carrier containing a physiologically acceptable agent depends, for example, on the route of administration of the composition. The preparation or pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) may also be a liposome or other polymeric matrix, and the compound of the present invention may be incorporated therein, for example. Liposomes, which may contain, for example, phospholipids or other lipids, are non-toxic, physiologically acceptable and metabolizable carriers for which preparation and administration are relatively simple.

[0088] As used herein, the term "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms suitable for use in contact with the tissues of humans and animals within the scope of sound medical judgment, without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0089] As used herein, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can function as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch and potato starch; (3) celluloses and their derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (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, corn oil, and soybean oil; (10) glycols such as propylene glycol; (11) polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances used in pharmaceutical formulations.

[0090] The pharmaceutical composition (preparation) can be administered to a subject by any of several routes of administration, including, for example, oral (such as aqueous or non-aqueous solutions, aqueous drugs such as those in aqueous or non-aqueous solutions, or suspensions, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue); absorption through the oral mucosa (such as sublingual); subcutaneous; transdermal (such as as a patch applied to the skin); and topical (such as as a cream, ointment, or spray applied to the skin). The compound may also be formulated for inhalation. In certain embodiments, the compound is simply dissolved or suspended in sterile water. Details regarding suitable routes of administration and the compositions suitable therefor can be found, for example, in U.S. Pat. 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 cited therein.

[0091] The formulations can conveniently be presented in unit dosage form and can be prepared by any method well known in the art of pharmacy. 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 to be treated and the particular mode 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 a therapeutic effect. Generally, out of 100 percent, this amount is an active ingredient in the range of about 1 percent to about 99 percent, preferably about 5 percent to about 70 percent, and most preferably about 10 percent to about 30 percent.

[0092] The methods of preparing these formulations and compositions include the step of associating an active compound, such as a compound of the invention, with a carrier and optionally one or more auxiliary components. Generally, the formulations are prepared by uniformly and intimately associating the compound of the invention with a liquid carrier or a micronized solid carrier or both, and then, if necessary, shaping the product.

[0093] The pharmaceutical preparations of the present invention suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (flavored bases, usually using sucrose and acacia or tragacanth), lyophilized products, powders, granules, or as solutions or suspensions in aqueous or non-aqueous liquids, or as water-in-oil or oil-in-water liquid emulsions, or as elixirs or syrups, or as troches (using inert bases such as gelatin and glycerin or sucrose and acacia), and / or as mouthwashes, etc., each containing a predetermined amount of the compound of the present invention as an active ingredient. The composition or compound may also be administered as a bolus, a lickable preparation, or a paste.

[0094] To prepare solid dosage forms for oral administration (including capsules (including sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, and granules, etc.), the active ingredient is one or more pharmaceutically acceptable carriers such as sodium citrate or dicalcium phosphate, and / or the following: (1) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders such as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants such as glycerol; (4) disintegrants such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retardants such as paraffin; (6) absorption promoters such as quaternary ammonium compounds; (7) wetting agents such as cetyl alcohol and glycerol monostearate; (8) absorbents such as kaolin and bentonite clay; (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) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets, and pills, the pharmaceutical composition may also contain a buffering agent. Solid compositions of the same type can also be used as fillers in soft and hard filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol.

[0095] Tablets can be made, optionally with one or more auxiliary components, by compression or molding. Compressed tablets can be prepared using binders (e.g., gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), surfactants, or dispersants. Molded tablets can be made by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0096] Tablets and other solid dosage forms of pharmaceutical compositions (such as dragees, capsules (including sprinkle capsules and gelatin capsules), pills, and granules) can optionally be scored or prepared using coatings and shells such as enteric coatings and other coatings well-known in the pharmaceutical formulation art. They can also be formulated to provide sustained or controlled release of the active ingredient therein, for example, using different proportions of hydroxypropyl methylcellulose, other polymer matrices, liposomes, and / or microparticles to provide a desired release profile. They can be sterilized, for example, by filtration through a bacteria-retaining filter or by incorporating a sterile agent in the form of a sterile solid composition that can be dissolved in sterile water or some other injectable sterile medium immediately before use. These compositions can also optionally contain an opacifying agent and can be of the type that releases the active ingredient(s) only to or preferentially to a specific part of the gastrointestinal tract in an optional delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient can also be in microencapsulated form, if appropriate, together with one or more of the excipients described above.

[0097] Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, lyophilized products for reconstitution, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, liquid dosage forms can contain, for example, inert diluents commonly used in the art (such as water or other solvents), cyclodextrins and their derivatives, solubilizing and emulsifying agents (such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (specifically, cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan), and mixtures thereof.

[0098] In addition to the inert diluent, the oral composition may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents, coloring agents, fragrances, and preservatives.

[0099] In addition to the active compound, the suspension may contain suspending agents (such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol, and sorbitan esters, etc.), microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, and tragacanth, and mixtures thereof.

[0100] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compound can be mixed under sterile conditions with a pharmaceutically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0101] Ointments, pastes, creams, and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof, in addition to the active compound.

[0102] Powders and sprays may contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures of these substances, in addition to the active compound. In addition, sprays may contain conventional propellants such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons (butane and propane, etc.).

[0103] The transdermal patch has the additional advantage of providing controlled delivery of the compounds of the present invention to the body. Such dosage forms can be prepared by dissolving or dispersing the active compound in a suitable medium. Absorption enhancers can also be used to increase the flow of the compound across the skin. The rate of such flow can be controlled either by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0104] As used herein, the terms "parenteral administration" and "administered parenterally" mean modes of administration other than enteral and topical administration usually by injection, including, without limitation, intravenous, intramuscular, intraarterial, intracavitary, intrasynovial, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration may contain one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that may be reconstituted into injectable sterile solutions or dispersions immediately before use, which may contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0105] Examples of suitable aqueous and non-aqueous carriers that may be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, and polyethylene glycol), and suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters (such as ethyl oleate). Suitable fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the particle size required in the case of dispersions, and by the use of surfactants.

[0106] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of the action of microorganisms can be ensured by including various antibacterial and antifungal agents, such as parabens, chlorobutanol, and phenolsorbic acid. It may also be desirable to include isotonic agents such as sugars and sodium chloride in the composition. In addition, the prolongation of the absorption of injectable pharmaceutical forms can be brought about by including agents that delay absorption, such as aluminum monostearate and gelatin.

[0107] In some cases, it is desirable to slow the absorption of drugs from subcutaneous or intramuscular injections in order to prolong the effect of the drug. This can be achieved by the use of liquid suspensions of crystalline or amorphous materials with poor water solubility. Thus, the rate of drug absorption depends on its dissolution rate, which in turn can depend on crystal size and crystal form. Alternatively, delayed absorption of parenterally administered drug forms is achieved by dissolving or suspending the drug in an oily vehicle.

[0108] Injectable depot forms are prepared by forming a microencapsulated matrix of the subject compound in a biodegradable polymer such as polylactic acid - polyglycolide. Depending on the ratio of drug to polymer and the nature of the particular polymer used, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Injectable depot formulations are also prepared by encapsulating the drug in liposomes or microemulsions that are compatible with body tissues.

[0109] For use in the methods of the present invention, the active compound can be provided, by itself or in combination with, for example, a pharmaceutically acceptable carrier, as a pharmaceutical composition containing from 0.1 to 99.5% (more preferably from 0.5 to 90%) active ingredient.

[0110] The delivery method can also be provided by rechargeable or biodegradable devices. In recent years, various sustained-release polymer devices for the controlled delivery of drugs, including proteinaceous biological agents, have been developed and tested in vivo. Various biocompatible polymers (including hydrogels), both biodegradable and non-biodegradable, can be used to form implants for the sustained release of compounds at specific target sites.

[0111] The actual dosage level of the active ingredient in the pharmaceutical composition can be varied to obtain an amount of the active ingredient that is effective in achieving the desired therapeutic response for a particular patient, composition, and mode of administration without being toxic to the patient.

[0112] The selection of the dosage level will depend on a variety of factors including the activity of the specific compound or combination of compounds, or esters, salts, or amides thereof, the route of administration, the time of administration, the rate of excretion of the specific compound(s) used, the duration of the treatment, other drugs, compounds, and / or materials used in combination with the specific compound(s) used, the age, sex, weight, medical condition, general health, and previous medical history of the patient being treated, as well as similar factors well known in the art of medicine.

[0113] A physician or veterinarian who is skilled in the art can readily determine and prescribe a therapeutically effective amount of the required pharmaceutical composition. For example, a physician or veterinarian can initiate the dosage of the pharmaceutical composition or compound at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. A "therapeutically effective amount" means an amount of the compound sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of a compound will vary depending on the weight, sex, age, and medical history of the subject. Other factors that can affect the effective amount can include, but are not limited to, the severity of the patient's medical condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent administered in combination with the compounds of the present invention. Larger total dosages may be delivered by multiple administrations of the agent. Methods for determining efficacy and dosage are known to those of skill in the art (Isselbacher et al. (1996) Harrison’s Principles of Internal Medicine 13ed.,1814-1882, which is incorporated herein by reference).

[0114] Generally, a suitable daily dosage of the active compound used in the compositions and methods of the present invention is the amount of the compound that is the lowest dosage effective to produce a therapeutic effect. Such effective dosages will generally depend on the factors described above.

[0115] If desired, the effective daily dosage of the active compound can be administered as one, two, three, four, five, six, or more sub-dosages, optionally in unit dosage forms, separately at appropriate intervals throughout the day. In certain embodiments of the present invention, the active compound can be administered one or two times a day. In a preferred embodiment, the active compound will be administered once a day.

[0116] The patient to be treated is any animal in need of treatment, including primates (specifically, humans), and other mammals (such as horses, cows, pigs, sheep, cats, and dogs), poultry, and common pets.

[0117] In certain embodiments, the compounds of the invention may be used alone or administered in combination with another type of therapeutic agent.

[0118] The present disclosure includes the use of pharmaceutically acceptable salts of the compounds of the invention in the compositions and methods of the invention. In certain embodiments, contemplated salts of the invention include, but are not limited to, alkyl, dialkyl, trialkyl, or tetra-alkylammonium salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, L-arginine, benethamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, Na, Ca, K, Mg, Zn, or other metal salts.In certain embodiments, the salts contemplated by the present invention include, but are not limited to, acid salts of 1-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, 1-ascorbic acid, 1-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d-glucoheptonic acid, d-gluconic acid, d-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, 1-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, pamoic acid, phosphoric acid, propionic acid, 1-pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, 1-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenic acid.

[0119] Pharmaceutically acceptable acid addition salts may also exist, for example, as various solvates with solvents such as water, methanol, ethanol, and dimethylformamide. Mixtures of such solvates may also be prepared. The source of such solvates may be derived from the crystallization solvent, inherent in the preparation or crystallization solvent, or accidentally present in such solvents.

[0120] Wetting agents, emulsifying agents, and lubricants (such as sodium lauryl sulfate and magnesium stearate), as well as coloring agents, releasing agents, coating agents, sweetening agents, flavoring agents and fragrances, preservatives, and antioxidants may also be present in the composition.

[0121] Examples of pharmaceutically acceptable antioxidants include (1) water-soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, and sodium sulfite; (2) oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, and alpha-tocopherol; and (3) metal-chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, and phosphoric acid.

[0122] Definitions Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings commonly understood by one of ordinary skill in the art. In general, the nomenclature and techniques described herein in connection with chemistry, cell and tissue culture, molecular biology, cell biology and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics, and protein and nucleic acid chemistry are well known and commonly used in the art.

[0123] Unless otherwise indicated, the methods and techniques of the present disclosure are generally carried out according to conventional methods well known in the art and as described in various general and more specific references cited and discussed throughout this specification. See, for example, “Principles of Neural Science”, McGraw-Hill Medical, New York, N.Y. (2000), Motulsky, “Intuitive Biostatistics”, Oxford University Press, Inc. (1995), Lodish et al., “Molecular Cell Biology, 4th ed.”, W.H. Freeman & Co., New York (2000), Griffiths et al., “Introduction to Genetic Analysis, 7th ed.”, W.H. Freeman & Co., N.Y. (1999), and Gilbert et al., “Developmental Biology, 6th ed.”, Sinauer Associates, Inc., Sunderland, MA (2000).

[0124] Unless otherwise defined herein, chemical terms used herein are used according to their conventional usage in the art as exemplified by “The McGraw-Hill Dictionary of Chemical Terms”, Parker S., Ed., McGraw-Hill, San Francisco, C.A. (1985).

[0125] All of the above, as well as any other publications, patents, and published patent applications referenced in this application, are specifically incorporated herein by reference. In case of conflict, the present specification, including its specific definitions, will control.

[0126] As used herein, the term "agent" refers to a chemical compound (organic or inorganic compound, mixture of chemical compounds, etc.), biological macromolecule (nucleic acid, antibody (including not only a part thereof but also humanized, chimeric, and human antibodies as well as monoclonal antibodies), protein or a part thereof (e.g., peptide, lipid, carbohydrate), or an extract made from biological materials (such as bacteria, plants, fungi, or animal (especially mammalian) cells or tissues). Agents include, for example, agents whose structure is known and agents whose structure is not known. By virtue of their ability to inhibit AR or promote AR degradation, such agents may be suitable as "therapeutic agents" in the methods and compositions of the present disclosure.

[0127] The terms "patient", "subject", or "individual" are used interchangeably and refer to either a human or a non-human animal. These terms include mammals such as humans, primates, livestock (including cows, pigs, etc.), companion animals (e.g., dogs, cats, etc.), and rodents (e.g., mice and rats).

[0128] "Treating" a condition or patient refers to taking steps to obtain a beneficial or desired result, including clinical outcomes. As used herein and as is well understood in the art, "treatment" is an approach to obtain a beneficial or desired result, including clinical outcomes. Beneficial or desired clinical outcomes can include, but are not limited to, reduction or alleviation of one or more symptoms or conditions, attenuation of the degree of the disease, a stabilized (i.e., not worsening) disease state, prevention of the spread of the disease, delay or slowing of disease progression, remission or alleviation of the disease state, and (whether partial or complete) cure. "Treatment" can also mean extending the survival period compared to the expected survival period if the treatment had not been received.

[0129] The term "preventing" is recognized in the art and, when used in connection with a condition such as local recurrence (e.g., pain), a disease such as cancer, a complex syndrome such as heart failure, or any other medical condition, is well understood in the art and includes administration of a composition that reduces the frequency of symptoms of the medical condition in a subject or delays its onset, as compared to a subject not receiving the composition. Thus, prevention of cancer, for example, includes reducing the number of detectable cancerous growths in a population of patients receiving preventive treatment, by a statistically and / or clinically significant amount, compared to an untreated control population, and / or delaying the appearance of detectable cancerous growths in the treated population, as compared to the untreated control population.

[0130] "Administering" or "administration" of a substance, compound, or agent to a subject can be effected using one of a variety of methods known to those of skill in the art. For example, a compound or agent can be administered intravenously, arterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, ocularly, sublingually, orally (by oral ingestion), intranasally (by inhalation), intraspinally, intracranially, and transdermally (e.g., by absorption through a skin conduit). A compound or agent can also be suitably introduced by a rechargeable or biodegradable polymer device or other device (e.g., a patch and a pump) or formulation that provides for extended release, sustained release, or slow release of the compound or agent. Administration can also be effected, for example, once, a plurality of times, and / or over one or more extended periods of time.

[0131] The appropriate method of administering a substance, compound, or agent to a subject will also depend, for example, on the age and / or physical condition of the subject, as well as the chemical and biological properties of the compound or agent (e.g., solubility, digestibility, bioavailability, stability, and toxicity). In some embodiments, a compound or agent is administered orally, for example, by oral ingestion to the subject. In some embodiments, a compound or agent administered orally is in an extended release or sustained release formulation or is administered using a device for such sustained release or extended release.

[0132] As used herein, the phrase "co - administration" refers to the administration of two or more different therapeutic agents in any form such that a second agent is administered while a previously administered therapeutic agent is still active in the body (e.g., the two agents are both active simultaneously in a patient, which may include a synergistic effect of the two agents). For example, different therapeutic compounds may be administered simultaneously or sequentially, either in the same formulation or in separate formulations. Thus, an individual receiving such treatment can benefit from the combined effect of the different therapeutic agents.

[0133] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or agent is the amount of the drug or agent that has the intended therapeutic effect upon administration to a subject. A complete therapeutic effect is not necessarily achieved by administration of a single dose and may occur only after administration of a series of doses. Thus, a therapeutically effective amount can be administered by one or more administrations. The exact effective amount required for a given subject will depend, for example, on the size, health, and age of the subject, as well as the nature and extent of the condition being treated (such as cancer or MDS). One of ordinary skill in the art can readily determine the effective amount for a given situation by conventional experimental methods.

[0134] The term "acyl" is recognized in the art and refers to a group represented by the general formula, hydrocarbyl C(O)-, preferably alkyl C(O)-.

[0135] The term "acylamino" is recognized in the art and refers to an amino group substituted with an acyl group and can be represented, for example, by the formula, hydrocarbyl C(O)NH-.

[0136] The term "acyloxy" is recognized in the art and refers to a group represented by the general formula, hydrocarbyl C(O)O-, preferably alkyl C(O)O-.

[0137] The term "alkoxy" refers to an alkyl group having an oxygen atom attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, and tert-butoxy.

[0138] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and can be represented by the general formula, alkyl-O-alkyl.

[0139] The term "alkyl" refers to a saturated aliphatic group including a straight-chain alkyl group, a branched-chain alkyl group, a cycloalkyl (alicyclic) group, an alkyl-substituted cycloalkyl group, and a cycloalkyl-substituted alkyl group. In a preferred embodiment, the straight-chain or branched-chain alkyl has 30 or fewer (e.g., C 1-30 , in the case of a branched chain C 3-30 ), and more preferably 20 or fewer carbon atoms in its backbone.

[0140] Furthermore, when used throughout this specification, examples, and claims, the term "alkyl" is intended to include both unsubstituted alkyl groups and substituted alkyl groups, the latter referring to an alkyl moiety having a substituent that replaces a hydrogen on one or more carbons of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl.

[0141] "C x-y " or "C x -C y ", when used in combination with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, means a group containing x to y carbons in the chain. C0 alkyl represents hydrogen when the group is in a terminal position and a bond when it is internal. A C 1-6 alkyl group contains, for example, 1 to 6 carbon atoms in the chain.

[0142] As used herein, the term "alkylamino" refers to an amino group substituted with at least one alkyl group.

[0143] 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-.

[0144] As used herein, the term "amide" refers to the group

[0145]

Chemical formula

[0146] The terms "amine" and "amino" are recognized in the art and include both unsubstituted and substituted amines, as well as their salts, for example,

[0147]

Chemical formula

[0148] As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group.

[0149] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group.

[0150] As used herein, the term "aryl" includes substituted or unsubstituted monocyclic aromatic groups in which each atom of the ring is carbon. Preferably, the ring is a 5- to 7-membered ring, more preferably a 6-membered ring. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, at least one of those rings being aromatic, for example, the other cyclic ring can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Aryl groups include benzene, naphthalene, phenanthrene, phenol, and aniline, among others.

[0151] The term "carbamate" is recognized in the art and refers to the group,

[0152] [Chemical formula] wherein R 9 and R 10 each independently represent hydrogen or a hydrocarbyl group.

[0153] As used herein, the term "carbocyclic alkyl" refers to an alkyl group substituted with a carbocyclic group.

[0154] As used herein, the terms "carbocyclic", "carbocyclyl", and "carbocyclic" refer to non-aromatic saturated or unsaturated rings in which each atom of the ring is carbon. Preferably, the carbocyclic ring contains 3 to 10 atoms, more preferably 5 to 7 atoms.

[0155] As used herein, the term "carbocyclic alkyl" refers to an alkyl group substituted with a carbocyclic group.

[0156] The term "carbonate" is recognized in the art and refers to the group, -OCO2-.

[0157] As used herein, the term "carboxy" refers to a group represented by the formula, -CO2H.

[0158] As used herein, the term "ester" refers to a group, -C(O)OR 9 wherein R 9 represents a hydrocarbyl group.

[0159] As used herein, the term "ether" refers to a hydrocarbyl group bonded through oxygen to another hydrocarbyl group. Thus, an ether substituent of a hydrocarbyl group can be hydrocarbyl-O-. The ether can be either symmetric or asymmetric. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. The ether includes an "alkoxyalkyl" group that can be represented by the general formula, alkyl-O-alkyl.

[0160] As used herein, the terms "halo" and "halogen" mean halogen and include chloro, fluoro, bromo, and iodo.

[0161] As used herein, the terms "heteroalkyl" and "heteroarylalkyl" refer to an alkyl group substituted with a heteroaryl group.

[0162] The terms "heteroaryl" and "heteroaryl" preferably include a substituted or unsubstituted aromatic monocyclic structure of 5- to 7-membered rings, more preferably 5- to 6-membered rings, and these ring structures contain at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heteroaryl" and "heteroaryl" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, and at least one of these rings is heteroaromatic. For example, the other cyclic ring can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine.

[0163] As used herein, the term "heteroatom" means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0164] As used herein, the term "heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclic group.

[0165] The terms "heterocyclyl", "heterocyclic", and "heterocyclic" preferably refer to a substituted or unsubstituted non-aromatic ring structure of 3- to 10-membered rings, more preferably 3- to 7-membered rings, and these ring structures contain at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, and at least one of these rings is heterocyclic. For example, the other cyclic ring can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactone, and lactam.

[0166] As used herein, the term "hydrocarbyl" refers to a group that is bonded through a carbon atom having no =O or =S substituents, typically having at least one carbon-hydrogen bond and primarily a carbon backbone, but optionally containing heteroatoms. Thus, even groups such as methyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered hydrocarbyl for the purposes of this application, while substituents such as acetyl (having an =O substituent on the bonded carbon) and ethoxy (bonded through oxygen rather than carbon) are not hydrocarbyl. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocyclic, heterocyclic, alkyl, alkenyl, alkynyl, and combinations thereof.

[0167] As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group.

[0168] When used in combination with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, the term "lower" means including groups having 10 or fewer, preferably 6 or fewer, atoms in the substituent. For example, "lower alkyl" refers to an alkyl group containing 10 or fewer, preferably 6 or fewer, carbon atoms. In certain embodiments, each of the acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents defined herein is a lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, whether they appear alone or in combination with other substituents, as in the listing of hydroxyalkyl and aralkyl (in which case, for example, when counting the carbon atoms in an alkyl substituent, the atoms in the aryl group are not counted).

[0169] The terms "policyclic", "polycyclic", and "polycyclic ring system" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl) where two or more atoms are common to two adjacent rings, e.g., the rings are "fused rings". Each of the rings of the polycyclic ring system may be substituted or unsubstituted. In certain embodiments, each ring of the polycyclic ring system contains from 3 to 10 atoms, preferably from 5 to 7 atoms, within the ring.

[0170] The term "sulfate" is recognized in the art and refers to the group, -OSO3H, or a pharmaceutically acceptable salt thereof.

[0171] The term "sulfonamide" is recognized in the art and has the general formula

[0172]

Chemical formula

[0173] The term "sulfoxide" is recognized in the art and refers to the group, -S(O)-.

[0174] The term "sulfonate" is recognized in the art and refers to the group, SO3H, or a pharmaceutically acceptable salt thereof.

[0175] The term "sulfone" is recognized in the art and refers to the group, -S(O)2-.

[0176] The term "substituted" refers to a moiety having a substituent that replaces a hydrogen on one or more carbons of a backbone. "Substitution" or "substituted with" is understood to include the implied conditions that such substitution follows the valences of the substituting atoms and substituents, and that the substitution results in a stable compound that does not readily undergo transformation, such as rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is intended to include all acceptable substituents of an organic compound. In a broad aspect, acceptable substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of an organic compound. Acceptable substituents may be one or more and may be the same or different for a given organic compound. For the purposes of the present invention, heteroatoms such as nitrogen may have hydrogen substituents and / or any acceptable substituents of the organic compounds described herein that satisfy the valences of the heteroatoms. Substituents may include any of the substituents described herein, for example, halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as thioester, thioacetate, or thiolate), alkoxyl, phosphoryl, phosphate, phosphonate, phosphinate, amino, amide, amidine, imine, cyano, nitro, azide, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamide, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. It will be understood by those skilled in the art that where appropriate, moieties substituted on a hydrocarbon chain may themselves be substituted.

[0177] As used herein, the term "thioalkyl" refers to an alkyl group substituted with a thiol group.

[0178] As used herein, the term "thioester" refers to the group, -C(O)SR 9 or -SC(O)R 9 and refers to wherein R 9 represents a hydrocarbyl.

[0179] As used herein, the term "thioether" corresponds to an ether in which oxygen is replaced by sulfur in the formula.

[0180] The term "urea" is recognized in the art and has the general formula

[0181] [Chemical formula] which can be represented by, wherein R 9 and R 10 each independently represent hydrogen or hydrocarbyl.

[0182] As used herein, the term "modulate" includes not only inhibiting or suppressing a function or activity (such as cell proliferation), but also enhancing a function or activity.

[0183] The phrase "pharmaceutically acceptable" is recognized in the art. In certain embodiments, this term refers to compositions, excipients, adjuvants, polymers, and other materials and / or dosage forms that are suitable for use in contact with human and animal tissues within the scope of sound medical judgment, without undue toxicity, irritation, allergic reaction, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0184] "Pharmaceutically acceptable salt" or "salt" is used herein to refer to acid addition salts or base addition salts that are suitable for or compatible with the treatment of a patient.

[0185] As used herein, "pharmaceutically acceptable acid addition salts" means non-toxic organic or inorganic salts of any basic 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 hydrogen 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). Either monoacid salts or diacid salts may be formed, and such salts can exist in hydrated, solvated, or substantially anhydrous forms. Generally, the acid addition salts of the compounds of Formula I or II are soluble in water and various hydrophilic organic solvents and generally demonstrate higher melting points compared to their free base forms. The selection of appropriate salts is known to those skilled in the art. For example, in the isolation of compounds of Formula I or II for use in the laboratory or for subsequent conversion to pharmaceutically acceptable acid addition salts, other pharmaceutically unacceptable salts, such as oxalic acid, may be used.

[0186] As used herein, "pharmaceutically acceptable base addition salts" means any non-toxic organic or inorganic base addition salts of any acid compound represented by Formula I or II, or any intermediate thereof. Exemplary inorganic bases that form suitable salts include lithium hydroxide, sodium, potassium, calcium, magnesium, or barium. Exemplary organic bases that form suitable salts include aliphatic, cycloaliphatic, or aromatic organic amines (such as methylamine, trimethylamine, and picoline), or ammonia. The selection of appropriate salts is known to those skilled in the art.

[0187] Many of the compounds useful in the methods and compositions of the present disclosure have at least one chiral center within their structure. This chiral center can exist in the R or S configuration, and the R and S notations are used in accordance with the rules described in Pure Appl. Chem. (1976), 45, 11-30. The present disclosure contemplates all stereoisomeric forms, such as enantiomeric and diastereomeric forms, of the compounds, salts, prodrugs, or mixtures thereof (including mixtures of all possible stereoisomers). See, for example, WO01 / 062726.

[0188] Furthermore, certain compounds containing an alkenyl group can exist as Z (cis) or E (trans) isomers. In each instance, the present disclosure includes both mixtures and the separate and individual isomers.

[0189] Some of the compounds may also exist in tautomeric forms. Although not explicitly shown in the formulas described herein, such forms are intended to be included within the scope of the present disclosure.

[0190] "Prodrug" or "pharmaceutically acceptable prodrug" refers to a compound that, after administration, is metabolized in the host, for example, hydrolyzed or oxidized, to form a compound of the present disclosure (e.g., a compound of formula I or II). Typical examples of prodrugs include compounds having a biologically labile group or a cleavable group (protecting group) on a functional moiety of the active compound. Prodrugs include compounds that can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated, acylated, deacylated, phosphorylated, or dephosphorylated to produce the active compound. Examples of prodrugs using esters or phosphoramidates as biologically labile groups or cleavable groups (protecting groups) are disclosed in U.S. Pat. Nos. 6,875,751, 7,585,851, and 7,964,580, the disclosures of which are incorporated herein by reference. The prodrugs of the present disclosure are metabolized to produce compounds of formula I or II. The present disclosure includes within its scope prodrugs of the compounds described herein. Conventional procedures for the selection and preparation of suitable prodrugs are described, for example, in "Design of Prodrugs" Ed. H. Bundgaard, Elsevier, 1985.

[0191] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, useful in the formulation of drugs for pharmaceutical or therapeutic use.

[0192] 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 often accompanies poor absorption. The LogS value is the unit stripped logarithm (base 10) of solubility measured in moles per liter.

Examples

[0193] Now that the present invention has been generally described, it will be more readily understood by reference to the following examples, which are included for the purpose of merely exemplifying specific aspects and embodiments of the present invention and are not intended to limit the present invention.

[0194] Example 1: Preparation of Exemplary Compounds

Chemical formula

[0195] To an ethanol solution (1 mL) of 1-phenyl-indole-3-carbaldehyde (1 mmol, 221 mg), 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 hours, and a yellow solid gradually precipitated. After completion of the reaction, ice-cold water (2 mL) was added to the reaction vial. The solid was separated by Buchner funnel filtration, washed with water (2 mL × 3), and dried to obtain the desired product. Yield: 95%, 300 mg.

[0196] To a solution of (E)-ethyl 2-cyano-3-(1-phenyl-1H-indol-3-yl)acrylate (0.32 mmol, 100 mg) in THF (2 mL) was added 0.5 N LiOH solution (3 equivalents, 0.6 mmol, 1.2 mL). The reaction mixture was stirred at 21 °C for 1 hour. After completion of the reaction as indicated by TLC, THF was evaporated. Concentrated HCl was added dropwise to oxidize the reaction mixture until the pH was less than 1, during which a yellow solid precipitated. Ice-cold water (5 mL) was added to the reaction mixture, and the solid was separated by Buchner funnel filtration and washed with water (5 mL × 3). After drying under vacuum, the solid was washed 5 - 10 times with 2 mL of a solvent mixture (hexane / EtOAc = 5:1) and monitored by TLC until non-polar impurities disappeared (the non-polar compounds were retro-aldol condensation products that could be recovered from the filtrate). Finally, the purity of the product was confirmed by NMR. Yield: 65%, 60 mg.

[0197] (E)-2-Cyano-3-(1-phenyl-1H-indol-3-yl)acrylic acid (JXL001 / UK5099) 1 H NMR (500 MHz, DMSO-d6) δ 8.59 (s, 1H), 8.56 (s, 1H), 8.06 (m, 1H), 7.65 (m, 4H), 7.53 (m, 2H), 7.34 (m, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.5, 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.

[0198]

Chemical Structure

[0199] The following compounds, JXL002, JXL003, JXL004, JXL005, JXL006, JXL007, JXL012, JXL013, JXL014, JXL021, JXL025, JXL026, JXL027, JXL028, JXL029, JXL035, JXL093 were synthesized by a route similar to the route described for JXL001.

[0200] [Chemical formula] (E)-2-Cyano-3-(1H-indol-3-yl)acrylic acid (JXL002) 1 H NMR (500 MHz, DMSO-d6) δ 12.48 (s, 1H), 8.51 (s, 1H), 8.49 (s, 1H), 7.91 (d, J = 6.5 Hz, 1H), 7.53 (d, J = 7.0 Hz, 1H), 7.23 (m, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 165.0, 146.5, 136.5, 132.4, 127.3, 123.9, 122.4, 118.9, 118.8, 113.2, 110.2, 94.0.

[0201] [Chemical formula] Ethyl (E)-2-cyano-3-(1H-indol-3-yl)acrylate 1 H NMR (500 MHz, CDCl3) δ 12.55 (s, 1H), 8.53 (s, 1H), 8.52 (s, 1H), 7.92 (d, J = 7.6 Hz, 1H), 7.53 (d, J = 7.8 Hz, 1H), 7.26 (app.t, J = 7.4 Hz, 1H), 7.22 (app.t, J = 7.4 Hz, 1H), 4.24 (q, J = 7.0 Hz, 2H), 1.26 (t, J = 7.0 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ 163.6, 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.

[0202] [Chemistry] Ethyl (E)-2-cyano-3-(1-phenyl-1H-indol-3-yl)acrylate (JXL004) 1 H NMR (500 MHz, CDCl3) δ 8.71 (s, 1H), 8.66 (s, 1H), 7.90 (d, J = 7.2 Hz, 1H), 7.54 (m, 6H), 7.36 (m, 2H), 4.39 (q, J = 7.1 Hz, 2H), 1.42 (t, J = 7.1 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ 163.6, 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.

[0203] [Chemistry] Ethyl (E)-3-(6-chloro-1-phenyl-1H-indol-3-yl)-2-cyanoacrylate (JXL005) 1 H NMR (500 MHz, CDCl3) δ 8.67 (s, 1H), 8.58 (s, 1H), 7.81 (d, J = 8.5 Hz, 1H), 7.60 (m, 2H), 7.52 (m, 4H), 7.34 (d, J = 8.4 Hz, 1H), 4.39 (q, J = 7.1 Hz, 2H), 1.41 (t, J = 7.1 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ 163.4, 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.

[0204] [Chemistry] Ethyl (E)-2-cyano-3-(1-(2-methoxyphenyl)-1H-indol-3-yl)acrylate (JXL006) 1 H NMR (500 MHz, CDCl3) δ 8.67 (s, 1H), 8.66 (s, 1H), 7.89 (d, J = 7.8 Hz, 1H), 7.49 (app.t, J = 8.6 Hz, 1H), 7.41 (d, J = 7.6 Hz, 1H), 7.35 (app.t, J = 7.3 Hz, 1H), 7.30 (t, J = 7.5 Hz, 1H), 7.23 (d, J = 8.1 Hz, 1H), 7.13 (m, 2H), 4.39 (q, J = 7.1 Hz, 2H), 3.81 (s, 3H), 1.41 (t, J = 7.1 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ 163.8, 154.2, 146.0, 137.2, 135.2, 130.3, 127.8, 126.0, 124.0, 122.6, 120.8, 118.2, 118.0, 112.3, 111.8, 111.0, 94.7, 61.8, 55.7, 14.3.

[0205]

Chem.

[0206] [Chemistry] (E)-2-Cyano-3-(1-(4-methoxyphenyl)-1H-indol-3-yl)acrylic acid (JXL012) 1 H NMR (500 MHz, DMSO-d6) δ 8.54 (s, 1H), 8.52 (s, 1H), 8.05 (d, J = 7.7 Hz, 1H), 7.58 (app.d, J = 8.7 Hz, 2H), 7.44 (m, 1H), 7.33 (m, 2H), 7.16 (app.d, J = 8.7 Hz, 2H), 3.82 (s, 3H). 13 C NMR (126 MHz, DMSO-d6) δ 164.7, 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.

[0207] [Chemistry] (E)-3-(6-Chloro-1-phenyl-1H-indol-3-yl)-2-cyanoacrylic acid (JXL013) 1 H NMR (500 MHz, DMSO-d6) δ 13.58 (br.s, 1H), 8.59 (s, 1H), 8.54 (s, 1H), 8.11 (d, J = 7.5 Hz, 1H), 7.67 (m, 4H), 7.53 (m, 2H), 7.35 (d, J = 7.5 Hz, 1H). 13 C NMR (126 MHz, DMSO-d6) δ 164.3, 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.

[0208] [Chemistry] (E)-2-Cyano-3-(1-(2-methoxyphenyl)-1H-indol-3-yl)acrylic acid (JXL014) 1 H NMR (500 MHz, DMSO-d6) δ 8.55 (s, 1H), 8.47 (s, 1H), 8.02 (d, J = 7.4 Hz, 1H), 7.54 (m, 2H), 7.31 (m, 3H), 7.15 (m, 2H), 3.74 (s, 3H). 13 C NMR (126 MHz, DMSO-d6) δ 164.6, 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.

[0209]

Chem.

[0210]

Chem.

[0211]

Chem.

[0212]

Chem.

[0213]

Chem.

[0214]

Chem.

[0215]

Chem.

[0216]

Chem.

[0217] Details of the experiment for the synthesis of JXL020

Chemical formula

[0218] To a solution of 1-(3,5-bis(trifluoromethyl)benzyl)-1H-indole-3-carbaldehyde (1 mmol, 371 mg) in ethanol (1 mL) were added ethyl 2-cyanoacetate (1.3 equiv, 1.3 mmol, 140 μL) and L-proline (40 mol%, 0.4 mmol, 58 mg). The reaction mixture was stirred at 21 °C for 12 h, and a yellow solid gradually precipitated. After completion of the reaction, ice-cold water (2 mL) was added to the reaction vial. The solid was separated by filtration through a Buchner funnel, washed with water (2 mL × 3), and dried to give the desired product. Yield: 93%, 433 mg.

[0219] To a solution of (E)-ethyl 3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyanoacrylate (0.21 mmol, 100 mg) in THF (2 mL) was added 0.5 N LiOH solution (3 equiv, 0.4 mmol, 0.8 mL). The reaction mixture was stirred at 21 °C for 1 h. After completion of the reaction as indicated by TLC, THF was evaporated. Concentrated HCl was added dropwise to acidify the reaction mixture until the pH was less than 1, during which 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 drying under vacuum, the solid was washed 5 - 10 times with 2 mL of a solvent mixture (hexane / EtOAc = 5:1) and monitored by TLC until the nonpolar impurities disappeared (the nonpolar compounds were retro-aldol condensation products that could be recovered from the filtrate). Finally, the purity of the product was confirmed by NMR. Yield: 55%, 52 mg.

[0220] (E)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyanoacrylic acid (JXL020) 1 H NMR (500 MHz, DMSO-d6) δ 13.37 (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). 1313C NMR (126 MHz, DMSO-d6) δ 164.7, 145.7, 140.3, 136.3, 134.8, 131.1, 130.8 (q, J = 31.1 Hz), 128.9, 128.7, 127.9, 124.8, 124.3, 122.9 (q, J = 273.4 Hz), 122.2, 119.3, 118.3, 95.6, 49.2.

[0221] By a route similar to the route described for JXL020, the following compounds, JXL008, JXL009, JXL010, JXL011, JXL015, JXL016, JXL017, JXL018, JXL019, JXL036, JXL037, JXL038, JXL039, JXL040, JXL041, JXL050, JXL051, JXL052, JXL053, JXL054, JXL055, JXL56, JXL057, JXL058, JXL059, JXL060, JXL061, JXL062, JXL063, JXL064, JXL065, JXL066, JXL068, JXL069, JXL072, JXL073, JXL076, JXL077, JXL078, JXL081, JXL082, JXL087, JXL088, JXL089, JXL090, JXL091 were synthesized.

[0222] [Chemical formula] Ethyl (E)-2-cyano-3-(1-(4-fluorobenzyl)-1H-indol-3-yl)acrylate (JXL008) 1 1H NMR (500 MHz, CDCl3) δ 8.60 (app.s, 2H), 7.85 (d, J = 6.8 Hz, 1H), 7.32 (m, 3H), 7.15 (m, 2H), 7.03 (app.t, 2H), 5.39 (s, 2H), 4.37 (q, J = 7.1 Hz, 2H), 1.40 (t, J = 7.1 Hz, 3H). 13 13C NMR (126 MHz, CDCl3) δ 163.7, 162.5 (d, J c-f= 247.7 Hz), 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 = 21.9 Hz), 110.9, 110.4, 94.6, 61.9, 50.7, 14.2.

[0223]

Chem.

[0224]

Chem.

[0225]

Chemical

[0226] [Chemical formula] (E)-2-Cyano-3-(1-(4-fluorobenzyl)-1H-indol-3-yl)acrylic acid (JXL015) 1 H NMR (500 MHz, DMSO-d6) δ 8.64 (s, 1H), 8.46 (s, 1H), 7.93 (d, J = 7.1 Hz, 1H), 7.61 (d, J = 7.3 Hz, 1H), 7.33 (m, 2H), 7.26 (m, 2H), 7.16 (m, 2H), 5.60 (s, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.8, 162.0 (d, J c-f = 244.3 Hz), 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 = 21.7 Hz), 112.0, 109.8, 95.0, 49.6.

[0227] [Chemical formula] (E)-2-Cyano-3-(1-(3,4-difluorobenzyl)-1H-indol-3-yl)acrylic acid (JXL016) 1 H NMR (500 MHz, DMSO-d6) δ 13.34 (br.s, 1H), 8.62 (s, 1H), 8.47 (s, 1H), 7.94 (d, J = 7.3 Hz, 1H), 7.61 (d, J = 7.6 Hz, 1H), 7.40 (m, 2H), 7.27 (m, 2H), 7.10 (br.s, 1H), 5.61 (s, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.8, 149.7 (dd, J c-f = 253.3, 13.6 Hz), 149.4 (dd, J c-f = 246.3, 11.6 Hz), 145.7, 136.4, 134.7, 134.5, 128.0, 124.7 (dd, J c-f=5.9, 3.0 Hz), 124.2, 122.8, 119.2, 118.4, 118.3 (d, J c-f =17.0 Hz), 117.1 (d, J c-f =17.6 Hz), 112.0, 109.9, 95.0, 49.3.

[0228]

Chem.

[0229]

Chem.

[0230]

Chem.

[0231]

Chem.

[0232]

Chem.

[0233]

Chem.

[0234]

Chem.

[0235] [Chemical] (E)-2-Cyano-3-(1-(3,5-difluorobenzyl)-6-fluoro-1H-indol-3-yl)acrylic acid (JXL040) 1 H NMR (500 MHz, DMSO-d6) δ 8.65 (s, 1H), 8.46 (s, 1H), 8.00 (dd, J = 8.6, 5.2 Hz, 1H), 7.57 (d, J = 9.7 Hz, 1H), 7.15 (m, 2H), 7.03 (s, 1H), 7.02 (s, 1H), 5.62 (s, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.8, 163.0 (d, J c-f = 239.4 Hz), 162.9 (d, J c-f = 248.6 Hz), 160.4 (d, J c-f = 239.4 Hz), 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 = 25.2 Hz), 98.7 (d, J c-f = 26.5 Hz), 96.9, 49.6.

[0236] [Chemical] (E)-3-(1-(3,5-Bis(trifluoromethyl)benzyl)-6-fluoro-1H-indol-3-yl)-2-cyanoacrylic acid (JXL041) 1 H NMR (500 MHz, DMSO-d6) δ 8.74 (s, 1H), 8.48 (s, 1H), 8.06 (app.s, 3H), 8.01 (dd, J = 8.7, 5.1 Hz, 1H), 7.66 (dd, J = 9.8, 2.0 Hz, 1H), 7.13 (dt, J = 9.3, 2.1 Hz, 1H), 5.78 (s, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.7, 159.9 (d, J c-f=264.6 Hz), 145.8, 140.2, 136.8, 135.3, 131.1 (q, J c-f =33.3 Hz), 129.1, 124.5, 123.6 (q, J c-f =273.7 Hz), 122.5, 121.2, 118.4, 111.4 (d, J c-f =25.2 Hz), 110.4, 98.6 (d, J c-f =27.2 Hz), 96.7, 49.4.

[0237]

Chem.

[0238]

Chem.

[0239]

Chem.

[0240]

Chem.

[0241]

Chem.

[0242]

Chem.

[0243]

Chem.

[0244]

Chem.

[0245]

Chem.

[0246]

Chem.

[0247] [Chemical formula] (E)-3-(1-(3,5-Bis(trifluoromethyl)benzyl)-4-methoxy-1H-indol-3-yl)-2-cyanoacrylic acid (JXL060) 1 H NMR (500 MHz, DMSO-d6) δ 8.99 (s, 1H), 8.71 (s, 1H), 8.05 (s, 1H), 8.00 (s, 2H), 7.24 (app.s, 2H), 6.82 (d, J = 6.0 Hz, 1H), 5.81 (s, 2H), 3.92 (s, 3H). 13 C NMR (126 MHz, DMSO-d6) δ 165.0, 154.8, 148.6, 140.5, 137.8, 133.3, 131.0 (q, J c-f = 32.8 Hz), 128.9, 125.4, 123.6 (q, J c-f = 273.7 Hz), 122.5, 118.5, 116.8, 110.5, 105.0, 104.3, 95.3, 56.2, 49.5.

[0248] [Chemical formula] (E)-3-(1-(3,5-Bis(trifluoromethyl)benzyl)-5-bromo-1H-indol-3-yl)-2-cyanoacrylic acid (JXL061) 1 H NMR (500 MHz, DMSO-d6) δ 8.75 (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). 13 C NMR (126 MHz, DMSO-d6) δ 164.7, 145.3, 140.3, 135.5, 135.3, 131.1 (q, J c-f = 32.8 Hz), 129.8, 128.9, 127.0, 124.7, 123.6 (q, J c-f = 274.0 Hz), 122.5, 122.3, 118.5, 113.9, 109.9, 97.7, 49.5.

[0249] [Chemical formula] (E)-3-(1-(3,5-Bis(trifluoromethyl)benzyl)-6-bromo-1H-indol-3-yl)-2-cyanoacrylic acid (JXL062) 1 H NMR (500 MHz, DMSO-d6) δ 8.72 (s, 1H), 8.47 (s, 1H), 8.06 (s, 2H), 8.03 (s, 1H), 7.94 (d, J = 8.2 Hz, 1H), 7.39 (d, J = 7.9 Hz, 1H), 5.81 (s, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.7, 145.6, 140.2, 137.4, 135.3, 131.1 (q, J c-f = 32.8 Hz), 129.1, 127.0, 125.9, 123.6 (q, J c-f = 274.0 Hz), 122.5, 121.5, 118.2, 117.2, 114.8, 110.3, 96.9, 49.3.

[0250] [Chemical formula] (E)-3-(1-(3,5-Bis(trifluoromethyl)benzyl)-6-chloro-1H-indol-3-yl)-2-cyanoacrylic acid (JXL063) 1 H NMR (500 MHz, DMSO-d6) δ 8.75 (s, 1H), 8.47 (s, 1H), 8.05 (m, 3H), 8.00 (d, J = 7.4 Hz, 1H), 7.89 (s, 1H), 7.28 (d, J = 6.5 Hz, 1H), 5.81 (s, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.7, 145.6, 140.2, 137.0, 135.4, 131.1 (q, J c-f = 32.8 Hz), 129.2, 129.0, 126.7, 123.6 (q, J c-f=274.0 Hz), 123.3, 122.5, 121.2, 118.2, 111.9, 110.3, 97.0, 49.3.

[0251]

Chem.

[0252]

Chem.

[0253]

Chem.

[0254]

Chem.

[0255]

Chem.

[0256]

Chem.

[0257]

Chem.

[0258]

Chem.

[0259]

Chem.

[0260]

Chem.

[0261]

Chem.

[0262]

Chem.

[0263]

Chem.

[0264]

Chem.

[0265]

Chem.

[0266]

Chem.

[0267]

Chem.

[0268] Details of the experiment for the synthesis of JXL024

Chem.

[0269] (Z)-5-((1-Phenyl-1H-indol-3-yl)methylene)thiazolidine-2,4-dione (JXL024) 1 H NMR (500 MHz, DMSO-d6) δ 7.98 (m, 2H), 7.79 (s, 1H), 7.66 (app.d, J = 7.7 Hz, 2H), 7.62 (app.t, J = 7.7 Hz, 2H), 7.54 (d, J = 8.0 Hz, 1H), 7.49 (t, J = 7.1 Hz, 1H), 7.30 (m, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 172.5, 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.

[0270] The following compounds, JXL067, JXL070, JXL072, JXL074, JXL075, were synthesized by a route similar to the route described for JXL024.

[0271]

Chemical formula

[0272]

Chemical formula

[0273]

Chem.

[0274]

Chem.

[0275]

Chem.

[0276]

Chem.

[0277] Details of the experiment for the synthesis of JXL022 [Chemical formula] To a solution of 4-pyridinecarboxaldehyde (1 mmol, 107 mg) in ethanol (1 mL), ethyl 2-cyanoacetate (1.3 equiv, 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 completion of the reaction, ice-cold water (2 mL) was added to the reaction vial. 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 was used in the next step without further purification.

[0278] A solution of (E)-2-cyano-3-(pyridin-4-yl)acrylate (0.21 mmol, 42.4 mg) in THF (2 mL) was added to 0.5 N LiOH solution (3 equivalents, 0.4 mmol, 0.8 mL). The reaction mixture was stirred at 21 °C for 1 hour. After completion of the reaction as indicated by TLC, THF was evaporated. Concentrated HCl was added dropwise to oxidize the reaction mixture until the pH was less than 1, during which a yellow solid precipitated. Ice-cold water (5 mL) was added to the reaction mixture, and the solid was separated by Buchner funnel filtration and washed with water (5 mL × 3). After drying under vacuum, the solid was washed 5 - 10 times with 2 mL of a solvent mixture (hexane / EtOAc = 5:1) and monitored by TLC until non-polar impurities disappeared. Finally, the purity of the product was confirmed by NMR. Yield: 64%, 23.4 mg.

[0279] Ethyl (E)-2-cyano-3-(pyridin-4-yl)acrylate (JXL022) 1 H NMR (500 MHz, CDCl3) δ 8.81 (d, J = 5.2 Hz, 2H), 8.18 (s, 1H), 7.74 (d, J = 5.2 Hz, 2H), 4.41 (q, J = 7.1 Hz, 2H), 1.41 (t, J = 7.1 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ 161.2, 152.0, 151.0, 137.9, 123.2, 114.2, 108.2, 63.2, 14.0.

[0280] Compounds JXL030, JXL031, JXL032, JXL033, JXL034, JXL042, JXL43, JXL044, JXL045, JXL046, JXL047, JXL048, JXL049 were synthesized by a route similar to the route described for JXL022.

[0281]

Chemical Structure

[0282]

Chem.

[0283]

Chem.

[0284] [Chemical formula] (E)-2-Cyano-3-(4-(trifluoromethyl)phenyl)acrylic acid (JXL034) 1 H NMR (500 MHz, DMSO-d6) δ 8.39 (s, 1H), 8.17 (d, J = 7.7 Hz, 2H), 7.84 (d, J = 7.7 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 162.8, 152.6, 135.4, 133.1 (q, J c-f = 32.9 Hz), 131.0, 125.7, 123.7 (q, J c-f = 272.2 Hz), 114.8, 106.7.

[0285] [Chemical formula] (E)-2-Cyano-3-(3-fluoro-4-methylphenyl)acrylic acid (JXL042) 1 H NMR (500 MHz, DMSO-d6) δ 8.28 (s, 1H), 7.78 (m, 2H), 7.49 (t, J = 8.0 Hz, 1H), 2.30 (s, 3H). 13 C NMR (126 MHz, DMSO-d6) δ 163.6, 160.9 (d, J c-f = 244.6 Hz), 153.5, 133.0, 131.6 (d, J c-f = 7.5 Hz), 130.8 (d, J c-f = 17.6 Hz), 127.2, 117.9 (d, J c-f = 23.9 Hz), 116.4, 104.5, 15.0.

[0286] [Chemical formula] (E)-2-Cyano-3-(3,4-difluorophenyl)acrylic acid (JXL043) 11H NMR (500 MHz, DMSO-d6) δ 8.32 (s, 1H), 8.09 (d, J = 8.0 Hz, 1H), 7.94 (br.s, 1H), 7.67 (d, J = 8.8 Hz, 1H). 13 13C NMR (126 MHz, DMSO-d6) δ 163.3, 152.3, 152.2 (dd, J c-f = 255.4, 12.6 Hz), 149.9 (dd, J c-f = 248.2, 12.6 Hz), 129.8, 128.8, 120.1 (d, J c-f = 17.6 Hz), 119.1 (d, J c-f = 17.6 Hz), 116.3, 105.8.

[0287]

Chem.

[0288]

Chem.

[0289]

Chem.

[0290]

Chem.

[0291]

Chem.

[0292]

Chem.

[0293] Details of the experiment for the synthesis of JXL079

Chem.

[0294] A flask containing a stirring bar was sealed, evacuated, and refilled with argon three times. Anhydrous dichloromethane (20 mL) and DIBAL (1 M in hexane, 6 mmol, 6 mL) were added to the flask. The crude carboxylic acid (2 mmol, 544 mg) dissolved in dry dichloromethane (10 mL) was added to the reaction flask at -78 °C. After 2 hours, the reaction was complete as indicated by TLC, and the reaction was 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 flash column chromatography (hexane:ethyl acetate = 10:1) to obtain the desired product, 2-(3,5-bis(trifluoromethyl)phenyl)ethan-1-ol (yield: 90%, 464 mg).

[0295] To a dichloromethane solution (2 mL) of 2-(3,5-bis(trifluoromethyl)phenyl)ethan-1-ol (0.2 mmol, 51.6 mg), triethylamine (0.22 mmol, 31 μL) and methanesulfonyl chloride (MsCl, 0.2 mmol, 17 μL) were added at 0 °C. After stirring for 1 hour, the reaction was completed as indicated by TLC. The solvent was removed by passing air through the open flask. The residue was purified by flash column chromatography (hexane:ethyl acetate = 10:1) to obtain the desired product, 3,5-bis(trifluoromethyl)phenethyl methanesulfonate (yield: 82%, 55.1 mg).

[0296] A flask containing NaH (60%, 0.22 mmol, 8.8 mg) and a stir bar was sealed, evacuated, and refilled with argon three times. Anhydrous THF (3 mL) and a THF solution (2 mL) of tert-butyl (E)-2-cyano-3-(1H-indol-3-yl)acrylate (0.2 mmol, 53.6 mg) were added to the reaction flask. The reaction mixture was stirred for 30 minutes, and then 3,5-bis(trifluoromethyl)phenethyl methanesulfonate (0.164 mmol, 55.1 mg) in 2 mL of THF was added. The reaction was stirred for 24 hours 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 flash column chromatography (hexane:ethyl acetate = 10:1) to obtain the desired product, tert-butyl (E)-3-(1-(3,5-bis(trifluoromethyl)phenethyl)-1H-indol-3-yl)-2-cyanoacrylate (yield: 68%, 69 mg).

[0297] To a dichloromethane solution (2 mL) of methyl tert-butyl (E)-3-(1-(3,5-bis(trifluoromethyl)phenethyl)-1H-indol-3-yl)-2-cyanoacrylate (0.1 mmol, 50.8 mg), trifluoroacetic acid (3 equivalents, 0.3 mmol, 34 μL) was added. The reaction mixture was stirred at 21 °C for 30 minutes, and a yellow solid precipitated. After the reaction was completed as indicated by TLC, the reaction solvent was evaporated by passing air through the open flask. The solid was washed 5 - 10 times with 2 mL of a solvent mixture (hexane / EtOAc = 5:1) and monitored by TLC until all non-polar impurities disappeared. Finally, the purity of the product was confirmed by NMR. Yield: 87%, 39 mg.

[0298] (E)-3-(1-(3,5-Bis(trifluoromethyl)phenethyl)-1H-indol-3-yl)-2-cyanoacrylic acid (JXL079) 1 H NMR (500 MHz, CD3OD) δ 8.51 (s, 1H), 8.14 (s, 1H), 7.83 (d, J = 7.3 Hz, 1H), 7.74 (s, 1H), 7.52 (m, 3H), 7.31 (m, 2H), 4.64 (t, J = 6.4 Hz, 2H), 3.35 (t, J = 6.3 Hz, 2H). 13 C NMR (126 MHz, CD3OD) δ 165.3, 145.3, 141.1, 136.1, 133.6, 131.3 (q, J c-f = 32.8 Hz), 129.3, 128.3, 123.7, 123.3 (q, J c-f = 272.5 Hz), 122.3, 120.3, 118.1, 117.6, 110.6, 109.6, 94.1, 47.8, 35.0.

[0299] Experimental details of the synthesis of JXL080

Chemical Structure

[0300] Ethyl (E)-3-(1-(2-(3,5-bis(trifluoromethyl)phenyl)acetyl)-1H-indol-3-yl)-2-cyanoacrylate (JXL080) 1 H NMR (500 MHz, CDCl3) δ 8.90 (s, 1H), 8.52 (s, 1H), 8.49 (d, J = 8.1 Hz, 1H), 7.88 (s, 1H), 7.86 (s, 2H), 7.78 (d, J = 7.5 Hz, 1H), 7.47 (m, 2H), 4.50 (s, 2H), 4.41 (q, J = 7.1 Hz, 2H), 1.43 (t, J = 7.1 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ 168.0, 162.2, 144.2, 135.6, 134.4, 132.3 (q, J c-f = 33.6 Hz), 130.1, 129.8, 128.8, 127.8, 127.3, 125.5, 123.1 (q, J c-f = 273.3 Hz), 118.3, 117.1, 117.0, 115.9, 101.8, 62.8, 41.9, 14.3.

[0301] Details of the experiment for the synthesis of JXL083

Chemical formula

[0302] A 100 mL round-bottom flask equipped with a stir bar containing the carboxylic acid (5 mmol, 1935 mg) from the previous step was sealed, evacuated, and refilled with argon three times. To this flask was added dropwise 50 mL of dichloromethane and oxalyl chloride (25 mmol, 2.1 mL). The reaction mixture was stirred at 21 °C for 1.5 h. The reaction solvent was evaporated in vacuo and the resulting compound was used in the next step.

[0303] A 100 mL round-bottom flask equipped with a stir bar was sealed, evacuated, and refilled with argon three times. Diisopropylamine (5.5 mmol, 765 μL) and THF (10 mL) were added to the flask, and it was cooled to -78 °C. nBuLi (2.5 M in hexanes, 5 mmol, 2 mL) was slowly added to the flask. After stirring the mixture for 30 minutes, a THF solution (10 mL) of ethyl 2-cyanoacetate (5 mmol, 590 μL) was slowly added to the flask. After stirring the mixture for 1 hour, a THF solution (5 mL) of acyl chloride (5 mmol from the previous step) was slowly added to the reaction mixture. After 1 hour, the reaction was quenched by adding 1 M aqueous HCl (10 mL), and the mixture was extracted with ethyl acetate (10 mL × 3). The combined organic phases were dried over sodium sulfate and evaporated using a rotary evaporator. The solid was purified by flash column chromatography (hexanes:ethyl acetate = 10:1) to give the desired product, ethyl (Z)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyano-3-hydroxyacrylate (JXL083) (yield: 80%, 1.93 g). 1 H NMR (500 MHz, CDCl3) δ 14.61 (s, 1H), 8.67 (s, 1H), 8.31 (dd, J = 7.0 1.3 Hz, 1H), 7.84 (s, 1H), 7.57 (s, 2H), 7.33 (m, 2H), 7.21 (d, J = 7.4 Hz, 1H), 5.53 (s, 2H), 4.40 (q, J = 7.1 Hz, 2H), 1.41 (t, J = 7.1 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ 179.1, 172.2, 138.1, 136.0, 135.7, 132.7 (q, J c-f = 33.6 Hz), 126.9, 124.5, 123.7, 123.6, 122.6, 122.9 (q, J c-f = 273.4 Hz), 121.8, 118.3, 110.1, 109.6, 73.1, 62.3, 50.4, 14.3.

[0304] Details of the experiment for the synthesis of JXL084 [Chemistry] To a dichloromethane solution (10 mL) of JXL083 (0.5 mmol, 240 mg), pyridine (0.5 mmol, 40 μL) and acetyl chloride (1.0 mmol, 84 μL) were added. The reaction mixture was stirred for 1 hour, and TLC indicated that the reaction was complete. The reaction solvent was evaporated by passing air through the open flask. The residue was purified by flash column chromatography (hexane:ethyl acetate = 10:1) to obtain the desired product, ethyl (Z)-3-acetoxy-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-2-cyanoacrylate (JXL084) (yield: 86%, 225 mg). 1 H NMR (500 MHz, CDCl3) δ 8.56 (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 = 7.1 Hz, 2H), 2.48 (s, 3H), 1.36 (t, J = 7.1 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ 166.9, 161.4, 137.7, 136.2, 135.5, 132.8 (q, J c-f = 33.9 Hz), 127.0, 126.7, 124.6, 123.6, 122.8 (q, J c-f = 273.4 Hz), 122.7, 121.8, 117.7, 110.7, 109.8, 89.3, 61.8, 50.5, 29.7, 21.3, 14.2.

[0305] Details of the experiment for the synthesis of JXL085 [Chemistry] To a dichloromethane solution (10 mL) of JXL083 (0.5 mmol, 240 mg), triethylamine (1.0 mmol, 139.5 μL) and phosphoryl chloride (0.55 mmol, 520 μL) were added. The reaction mixture was stirred at reflux for 1 hour, and TLC indicated that the reaction was complete. The reaction solvent was evaporated by passing air through the open flask. The residue was purified by flash column chromatography (hexane:ethyl acetate = 10:1) to obtain the desired product, ethyl (Z)-3-(1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-3-chloro-2-cyanoacrylate (JXL085) (yield: 84%, 210 mg). 1 H NMR (500 MHz, CDCl3) δ 7.91 (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 = 7.1 Hz, 2H), 1.18 (t, J = 7.1 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ 161.0, 155.6, 138.1, 136.2, 135.4, 133.4, 132.7 (q, J c-f = 33.7 Hz), 127.1, 126.6, 124.5, 124.3, 122.8 (q, J c-f = 273.4 Hz), 121.5, 115.8, 122.1, 110.4, 102.5, 62.4, 50.0, 13.9.

[0306] Details of the experiment for the synthesis of JXL086

Chemical Structure

[0307] JXL095 was synthesized by a route similar to that of JXL086.

Chemical Structure

[0308] Details of the experiment for the synthesis of JXL096 [Chemical formula] (E)-(2-(1-(3,5-Bis(trifluoromethyl)benzyl)-1H-indol-3-yl)-1-cyanovinyl)phosphonic acid (JXL096) A dichloromethane solution (2 mL) of JXL086 (30 mg, 0.057 mmol) was cooled to 0 °C, and bromotrimethylsilane (40 μL, 0.3 mmol) was added dropwise under argon. The mixture was warmed to 21 °C and stirred for 12 hours. The solvent was evaporated under vacuum, and then the resulting residue was dissolved in methanol (2 mL). The mixture was stirred at 21 °C for 2 hours. Evaporation of all volatiles under vacuum gave phosphoric acid, JXL096 (yield: 92%, 25 mg). 11H NMR (500 MHz, CD3OD) δ 8.58 (s, 1H), 8.25 (d, J = 19.6 Hz, 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). 13 13C NMR (126 MHz, CD3OD) δ 146.7 (J c-p = 7.2 Hz), 140.2, 136.1, 132.1, 131.9 (q, J c-f = 33.7 Hz), 128.0, 127.2, 123.8, 122.3, 123.2 (q, J c-f = 273.4 Hz), 121.4, 118.2, 117.4 (d, J c-p = 11.3 Hz), 111.5 (J c-p = 18.4 Hz), 110.5, 94.6 (d, J c-p = 201.2 Hz), 49.1.

[0309] Details of the experiment for the synthesis of JXL092

Chemical Structure

[0310] (E)-3-(1-(3,5-Bis(trifluoromethyl)benzyl)-4-(methoxycarbonyl)-1H-indol-3-yl)-2-cyanoacrylic acid (JXL092) To a dichloromethane solution (2 mL) of methyl (E)-1-(3,5-bis(trifluoromethyl)benzyl)-3-(3-(tert-butoxy)-2-cyano-3-oxoprop-1-en-1-yl)-1H-indole-4-carboxylate (0.5 mmol, 276 mg), trifluoroacetic acid (3 eq, 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 completed as indicated by TLC, the reaction solvent was evaporated by passing air through the open flask. The solid was washed 5 - 10 times with 2 mL of a solvent mixture (hexane / EtOAc = 5:1) and monitored by TLC until all non-polar impurities disappeared. Finally, the purity of the product was confirmed by NMR. Yield: 90%, 223 mg. 1 H NMR (500 MHz, CDCl3) δ 9.09 (s, 1H), 8.58 (s, 1H), 7.75 (d, J = 7.5 Hz, 1H), 7.68 (s, 1H), 7.47 (s, 2H), 7.34 (d, J = 8.2 Hz, 1H), 7.20 (app.t, J = 7.9 Hz, 1H), 5.51 (s, 2H), 3.87 (s, 3H). 13 C NMR (126 MHz, CDCl3) δ 167.8, 165.1, 149.5, 138.2, 137.2, 134.8, 132.3 (q, J c-f = 33.7 Hz), 126.8, 126.2, 125.5, 124.8, 123.3, 122.8 (q, J c-f = 273.4 Hz), 122.3 118.2, 114.7, 110.9, 97.2, 52.4, 50.2.

[0311] Details of the experiment for the synthesis of JXL094

Chemical Structure

[0312] Additional exemplary compounds of the present invention can be prepared by a method similar to the above method.

[0313] Example 2: Treatment of epithelial cells with an exemplary compound To determine whether these compounds could promote cellular lactate production, we treated cultured epithelial cells with the compounds and measured the lactate levels in the culture medium using a Nova Biomedical BioProfile Basic Analyzer. Briefly, cultured epithelial cells were treated for 24 - 30 hours with DMSO, UK-5099 (also called JXL001), or some of the exemplary compounds disclosed herein, the lactate levels in the medium were measured, and normalized to cell number and duration of the experiment to obtain the cellular lactate production rate (nmol of lactate, per million cells, per hour).

[0314] The lactate production rates of the treated cells are shown in FIGS. 8, 9, and 12. As expected based on the present disclosure, since they are UK-5099 analogs, most of the novel compounds assayed increased lactate production. Further, the total cell numbers after treatment with UK-5099 analogs are shown in FIG. 13. For most of the compounds, the cells showed tolerance. Separate assays were performed to calculate the EC50 for some of the compounds shown in FIG. 10.

[0315] Example 3: In Vivo Testing of Exemplary Compounds To determine the effectiveness of the compounds on the hair cycle, mice were shaved on day 50 after birth and topically treated every other day for two weeks with the compounds disclosed herein (suspended in lotion), and photographs were taken. As seen in FIG. 11, all analogs that showed the ability to promote lactate production in the in vitro assay were also able to stimulate hair growth over two weeks.

[0316] Incorporation by Reference All publications and patents mentioned herein are hereby incorporated by reference as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.

[0317] Equivalents While specific embodiments of the invention are being considered, the above specification is illustrative and not restrictive. Many variations of the invention will be apparent to those skilled in the art upon consideration of this specification and the following claims. The full scope of the invention should be determined by reference to the claims, together with the full scope of their equivalents, and the specification in light of such variations. Examples of embodiments of the present invention can be cited as follows, for example. [Embodiment 1] A compound of formula I or II, [Chemical formula] wherein, each A is independently CH, CR 4 , or N, Y is carboxyl, ester, amide, or [Chemical formula] is, Z is CH, CR 4 , or N, R 2 is CN or carboxyl, R 3 is H, aryl, aralkyl, or aralkyl acyl, and is optionally substituted by one or more R 5 , each R 5 is independently selected from alkyl, alkoxy, or halo, R 4 each example of which is independently alkyl, carboxyl, halo, hydroxy, ester, or CN, R 7 is hydrogen, alkyl, halo, hydroxyl, alkoxy, or acyloxy, R 10 is hydrogen or alkyl, n is 0 to 4, said compound, or a pharmaceutically acceptable salt thereof. [Embodiment 2] The compound according to Embodiment 1, wherein Z is CH or N. [Embodiment 3] The compound is a compound of Formula III,

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Claims

1. A compound of formula I or II, 【Chemical 1】 wherein, Each A is independently CH, CR ４ , or N, and Y is carboxyl, ester, amide, or 【Chemical 2】 and, Z is CH, CR ４ , or N, and R ２ is CN or carboxyl, R ３ is H, aryl, aralkyl, or aralkylacyl and is optionally substituted by one or more R ５ each R ５ is independently selected from alkyl, alkoxy, or halo, R ４ each example of which is independently alkyl, carboxyl, halo, hydroxy, ester, or CN, R ７ is hydrogen, alkyl, halo, hydroxyl, alkoxy, or acyloxy, and R １０ is hydrogen or alkyl, and n is from 0 to 4, said compound, or a pharmaceutically acceptable salt thereof.

2. Z is CH or N, the compound according to Claim 1.

3. Said compound is a compound of formula III, [Chemical Formula 3] wherein, Y is carboxyl, ester, amide, or 【Chemical Formula 4】 and, R ２ is CN or carboxyl, R ３ is H, aryl, aralkyl, or aralkylacyl and is optionally substituted by one or more R ５ each R ５ is independently selected from alkyl, alkoxy, or halo, R ４ In each example of, independently, alkyl, carboxyl, halo, hydroxy, ester, or CN, R ６ is derived from H, alkyl, or cycloalkyl, R ７ is hydrogen, alkyl, halo, hydroxyl, alkoxy, or acyloxy, and R １０ is hydrogen or alkyl, R １１ is H or alkyl, n is from 0 to 4, said compound, or a pharmaceutically acceptable salt thereof, the compound according to Claim 1.

4. Y is 【Chemical Formula 5】 , the compound according to any one of Claims 1 to 3.

5. R １０ The compound according to claim 4, wherein R is H.

6. R １０ The compound according to claim 4, wherein R is alkyl (for example, ethyl).

7. Y is ester or amide, the compound according to any one of Claims 1 to 3.

8. R １１ The compound according to any one of claims 3 to 7, wherein R is alkyl (for example, methyl).

9. Said compound is a compound of formula V, VI, or VII, [Chemical Formula 6] wherein, Each A is independently CH, CR ４ , or N, and X is NR ６ or O, and R １ is H or lower alkyl, or R １ and R ６ or R １ and R ２ either together with the atoms separating them completes a heterocyclic ring, R ２ is CN or carboxyl, R ３ is H, aryl, aralkyl, or aralkylacyl and is optionally substituted by one or more R ５ each R ５ is independently selected from alkyl, alkoxy, or halo, R ４ each instance of which, independently, is alkyl, carboxyl, halo, hydroxy, or CN, R ６ is derived from H, alkyl, or cycloalkyl, R ７ is hydrogen, alkyl, halo, hydroxyl, alkoxy, or acyloxy, and n is from 0 to 4, said compound, or a pharmaceutically acceptable salt thereof, the compound according to Claim 1.

10. At least one A is N, the compound according to any one of Claims 1 to 9.

11. Exactly one A is N, the compound according to any one of Claims 1 to 10.

12. Said compound is a compound of formula Va, VIa, or VIIa, [Chemical Formula 7] wherein, X is NR ６ or O, and R １ is H or lower alkyl, R ２ is CN or carboxyl, or R １ and R ２ together with the atoms separating them complete a heterocyclic ring, R ３ is H, phenyl, or benzyl, and one or more R ５ are optionally substituted by, each R ５ is independently selected from alkyl, alkoxy, or halo, R ４ In each instance, is independently alkyl, carboxyl, halo, hydroxy, or CN, R ６ is selected from H, alkyl, or cycloalkyl, n is from 0 to 4, said compound, or a pharmaceutically acceptable salt thereof, the compound according to any one of Claims 1 to 11.

13. X is NH, the compound according to any one of Claims 8 to 12.

14. X is O, the compound according to any one of Claims 8 to 12.

15. R １ The compound according to any one of claims 9 to 12, wherein R is H.

16. R １ The compound according to any one of claims 9 to 12, wherein R is lower alkyl.

17. R １ and R ６ which together with the atoms separating them complete a heterocyclic ring (e.g., morpholinyl), a compound according to any one of claims 9 to 12.

18. R ６ The compound according to any one of claims 9 to 13, wherein R is hydrogen.

19. R ２ The compound according to any one of claims 9 to 18, wherein R is CN.

20. R ２ The compound according to any one of claims 9 to 18, wherein R is carboxyl.

21. R １ and R ２ together with the atoms separating them, complete a heterocyclyl selected from thiazolidine-2,4-dione-5-ylidene or 2-iminothiazolidin-4-one-5-ylidene, a compound according to any one of claims 9 to 13.

22. Said compound is a compound of formula Va, the compound according to any one of Claims 12 to 21.

23. Said compound is a compound of formula VIa, the compound according to any one of Claims 12 to 21.

24. R ３ The compound according to any one of claims 1 to 23, wherein R is H.

25. R ３ The compound according to any one of claims 1 to 23, wherein R is phenyl.

26. R ３ is phenyl and is substituted by one or more R ５ The compound according to any one of claims 1 to 23.

27. R ３ is replaced by one R ５ and R ５ is an alkoxy, the compound according to claim 26.

28. R ３ The compound according to claim 26 or 27, wherein R is aralkyl (for example, benzyl or phenethyl).

29. R ３ The compound according to claim 26 or 27, wherein R is aralkyl.

30. R ３ The compound according to claim 26 or 27, wherein R is aralkyl acyl (for example, phenylacetyl).

31. R ３ The compound according to claim 26 or 27, wherein R is benzyl.

32. R ３ is benzyl and is substituted by one or more R ５ groups, a compound according to claim 26 or 27

33. R ３ is aralkyl (e.g., benzyl or phenethyl) and is substituted by one or more R ５ as defined in claim 26 or 27.

34. R ３ wherein R is aralkyl acyl (for example, phenylacetyl) and is substituted by one or more R ５ (preferably on the phenyl ring), the compound according to claim 26 or 27.

35. R ３ is substituted by one or two R ５ and each R ５ is independently selected from fluoroalkyl or fluoro, the compound according to claim 32 or 33.

36. R ３ is replaced by two Rs ５ and each R ５ is trifluoromethyl, the compound according to claim 35.

37. Said compound is represented by formula Vb, the compound according to any one of Claims 9 to 36. 【Chemical 8】

38. n is 0, the compound according to any one of Claims 12 to 37.

39. The compound according to claim 38, wherein the compound is represented by formula Vc. 【Chemical Formula 9】

40. The compound according to any one of claims 12 to 37, wherein n is 1.

41. The compound according to claim 40, wherein the compound is represented by formula Vd. 【Chemical Formula 10】

42. The compound according to claim 40, wherein the compound is represented by formula Ve. 【Chemical 11】

43. R ４ The compound according to any one of claims 40 to 42, wherein R is selected from halo or haloalkyl.

44. R ４ The compound according to claim 43, wherein R is halo (e.g., chloro or bromo).

45. The compound according to any one of claims 9 to 21, wherein the compound is a compound of formula VI.

46. The compound according to any one of claims 9 to 21, wherein the compound is a compound of formula VIa.

47. The compound according to claim 45 or 46, wherein n is 0.

48. n is 2, and R ４ is selected from halo or haloalkyl, a compound according to claim 45 or 46.

49. The compound according to any one of claims 9 to 21, wherein the compound is a compound of formula VII.

50. The compound according to any one of claims 9 to 21, wherein the compound is a compound of formula VIIa.

51. R ７ The compound according to any one of the preceding claims, wherein R is hydrogen, hydroxyl, halo (for example, chloro), or acyloxy (for example, acetyloxy).

52. R ７ The compound according to claim 51, wherein R is hydroxyl, halo (e.g., chloro), or acyloxy (e.g., acetyloxy).

53. 【Fig. 12】 【Chem.】 【Chem.】 [Chemical] 【Chem.】 [Chemical] 【Chem.】 A compound selected from.

54. A pharmaceutical composition comprising the compound according to any one of the preceding claims and a pharmaceutically acceptable excipient.

55. The pharmaceutical composition according to claim 54, wherein the pharmaceutical composition is formulated for topical administration.

56. A method for enhancing intracellular lactic acid production, the method comprising contacting the cell with an MPO inhibitor such as an MPC inhibitor.

57. A method for enhancing intracellular lactic acid production, the method comprising contacting the cell with the compound or composition according to any one of claims 1 to 55.

58. The method according to claim 56 or 57, wherein the cell is a hair follicle stem cell.

59. A method for promoting hair growth, the method comprising administering to a patient the compound or composition according to any one of claims 1 to 55.

60. A method for treating a pathological condition or disorder affecting hair growth, the method comprising administering to a patient the compound or composition according to any one of claims 1 to 55.

61. The method according to claim 60, wherein the pathological condition or disorder is alopecia or hair loss.

62. A method for promoting hair growth, the method comprising administering (e.g., topically) an MPC inhibitor to a patient.

63. A method for treating a pathological condition or disorder affecting hair growth, the method comprising administering (e.g., topically) an MPC inhibitor to a patient.

64. The method according to claim 63, wherein the condition or disorder is alopecia or hair loss.

65. The method according to any one of claims 62 to 65, wherein the MPC inhibitor is the compound or composition according to any one of claims 1 to 55.

66. A method for promoting hair growth, comprising administering an MPO inhibitor to a patient (e.g., topically).

67. A method for treating a condition or disorder that affects hair growth, comprising administering an MPO inhibitor to a patient (e.g., topically).

68. The method according to claim 67, wherein the condition or disorder is alopecia or hair loss.

69. The method according to any one of claims 66 to 68, wherein the MPC inhibitor is the compound or composition according to any one of claims 1 to 55.

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