Malassezin and analogs thereof as skin brightening agents

Abstract

To provide compounds, compositions, and methods for brightening skin.SOLUTION: The present invention relates to compounds, compositions, and methods for brightening skin. The compounds, compositions, and methods of the present invention generally involve compounds produced by a Malassezia yeast, and chemical analogs thereof. In addition to skin brightening applications, the compounds, compositions, and methods of the present invention may be used to modulate melanocyte activity, induce melanocyte apoptosis, agonize an arylhydrocarbon receptor (AhR), improve hyperpigmentation caused by a hyperpigmentation disorder, and modulate melanin production, melanosome biogenesis, and melanosome transfer.SELECTED DRAWING: Figure 2C

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 62 / 306,468, filed March 10, 2016, the entire contents of which are incorporated by reference.

[0002] The present invention relates to chemical analogs of compounds produced by Malassezia yeast. The present invention includes compounds produced by Malassezia yeast as well as compositions containing chemical analogs of compounds produced by Malassezia yeast. Methods of using the compounds (including analogs thereof) and compositions of the present invention are also contemplated. [Background technology]

[0003] People around the world use skin lightening agents to achieve several cosmetic goals, such as obtaining anti-aging benefits, repairing sun damage, and meeting certain cultural standards of beauty. Many commercially available skin lightening products, while effective to varying degrees, contain harmful ingredients, some of which have been linked to cancer. Thus, there is a need for new skin lightening agents and formulations that exhibit a higher level of safety and / or efficacy than skin lightening agents currently on the market.

[0004] Malassezia is a genus of lipophilic yeasts commonly found in the normal flora of human skin and is responsible for several skin disorders, including black lichen (tinea versicolor), seborrheic dermatitis, and atopic dermatitis.

[0005] Black catfish is a non-contagious skin disease caused by Malassezia overgrowth, which locally alters pigmentation levels. Malassezia yeasts have two metabolic pathways for synthesizing melanin and tryptophan-derived indole pigments. The indole pigments contain malassedin, a tryptophan metabolite of Malassezia, which induces apoptosis of melanocytes and contributes to the depigmentation characteristic of Malassezia overgrowth.

[0006] The invention disclosed herein utilizes compounds produced by Malassezia yeast, including malassedins, and chemical analogs thereof, as the basis of safe and effective skin lightening compositions. Summary of the Invention [Means for solving the problem]

[0007] One embodiment of the present invention is a compound for skin whitening, which is a chemical analogue of the compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0008] Another embodiment of the present invention is a compound for inducing apoptosis of melanocytes, the compound being a chemical analogue of the compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0009] A further embodiment of the present invention is a compound for modulating melanocyte activity, which is a chemical analogue of the compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0010] An additional embodiment of the present invention is a compound for agonizing the aryl hydrocarbon receptor (AhR), which is a chemical analog of a compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0011] Another embodiment of the present invention is a compound for improving hyperpigmentation caused by hyperpigmentation disorders, the compound being a chemical analogue of the compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0012] A further embodiment of the present invention is a compound for modulating melanin production, which is a chemical analogue of the compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0013] An additional embodiment of the present invention is a compound for modulating melanosome biogenesis, which is a chemical analogue of the compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0014] Another embodiment of the present invention is a compound for modulating melanosome transport, which is a chemical analogue of the compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0015] A further embodiment of the invention is a composition comprising Malassezia yeast and a cosmetically or pharma- ceutically acceptable vehicle, diluent, or carrier.

[0016] An additional embodiment of the invention is a composition comprising a compound isolated or isolatable from Malassezia yeast and a cosmetically or pharma- ceutically acceptable vehicle, diluent, or carrier.

[0017] Another embodiment of the invention is a composition comprising any of the compounds disclosed herein, including analogs, and a cosmetically or pharma- ceutically acceptable vehicle, diluent, or carrier.

[0018] A further embodiment of the invention is a method for lightening the skin in a subject, the method comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0019] An additional embodiment of the invention is a method for inducing apoptosis of melanocytes in a subject, comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0020] Another embodiment of the invention is a method for modulating melanocyte activity in a subject, comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0021] A further embodiment of the invention is a method for agonizing the aryl hydrocarbon receptor (AhR) in a subject, comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0022] Additional embodiments of the present invention relate to hyperpigmentation caused by hyperpigmentation disorders. A method for improving hyperpigmentation caused by a hyperpigmentation disorder in a subject in need of such improvement, comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0023] Another embodiment of the invention is a method for modulating melanin production in a subject, comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0024] A further embodiment of the invention is a method for modulating melanosome biogenesis in a subject, comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0025] An additional embodiment of the invention is a method for modulating melanosome transport in a subject, comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0026] Another embodiment of the present invention is a compound having the formula (II): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl, and at least one of R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 is methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0027] A further embodiment of the present invention is a compound having the formula (III): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl; , at least one of R8, R9, and R10 is methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0028] An additional embodiment of the present invention is a compound for whitening the skin, the compound having the formula (II): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0029] Another embodiment of the present invention is a compound for skin whitening, the compound having the formula (III): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0030] A further embodiment of the present invention is a compound for inducing apoptosis of melanocytes, the compound having formula (II): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0031] An additional embodiment of the present invention is a compound for inducing apoptosis of melanocytes, the compound having the formula (III): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0032] Another embodiment of the present invention is a compound for agonizing the aryl hydrocarbon receptor (AhR), the compound having the formula (II): [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form or hydrate thereof; or a cosmetically or pharma- ceutically acceptable salt thereof.

[0033] A further embodiment of the present invention is a compound for agonizing the aryl hydrocarbon receptor (AhR), the compound having the formula (III): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0034] An additional embodiment of the present invention is a composition comprising a compound of formula (II): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl), or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof, and a cosmetically or pharma- ceutically acceptable vehicle, diluent, or carrier.

[0035] Another embodiment of the present invention is a composition comprising a compound of formula (III): [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof, and a cosmetically or pharma- ceutically acceptable vehicle, diluent, or carrier.

[0036] A further embodiment of the present invention is a method for whitening the skin in a subject, comprising administering to a subject a compound of formula (II): [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, contacting a subject with a compound having the structure:

[0037] An additional embodiment of the present invention is a method for lightening the skin in a subject, comprising: [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, to a subject.

[0038] Another embodiment of the present invention is a method for inducing apoptosis of melanocytes in a subject, the method comprising administering to a subject a compound of formula (II): [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, contacting a subject with a compound having the structure:

[0039] A further embodiment of the invention is a method for inducing apoptosis of melanocytes in a subject, comprising administering to a subject a compound of formula (III): [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, to a subject.

[0040] Another embodiment of the present invention is a method for agonizing the aryl hydrocarbon receptor (AhR) in a subject, the method comprising administering to a subject a compound of formula (II): [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, contacting a subject with a compound having the structure:

[0041] A further embodiment of the invention is a method for agonizing the aryl hydrocarbon receptor (AhR) in a subject, comprising administering to a subject a compound of formula (III): [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, to a subject.

[0042] The patent or patent application file contains at least one drawing executed in color. Copies of the patent or patent application publication with that drawing executed in color will be provided by the Office upon request and payment of the necessary fee. [Brief description of the drawings]

[0043] [Figure 1A] FIG. 1 is a schematic diagram of the component layers of skin, with the inset showing the cellular structure that comprises the epidermis and dermis. [Figure 1B] FIG. 1 is a schematic showing the potential mechanism of action of agents that cause depigmentation. [Figure 2A] 1 is a series of synthetic schemes for malasedin and malasedin derivatives (malasedin and indolo[3,2-b]carbazole). [Figure 2B] 1 is a series of synthetic schemes for malassezin derivatives (compounds I and IV). [Figure 2C] 1 is a series of synthesis schemes for a malassezin derivative (compound II). [Figure 3A] Figure 3A is a summary table showing EC50 values ​​for Annexin V induction for certain compounds of the invention in MeWo and WM115 cells, and Figures 3B-3M are line graphs showing the percentage of MeWo (Figures 3B-3G) or WM115 (Figures 3H-3M) cells labeled with Annexin V after exposure to various concentrations of exemplary compounds. [Figure 3B-D] Figure 3A is a summary table showing EC50 values ​​for Annexin V induction for certain compounds of the invention in MeWo and WM115 cells, and Figures 3B-3M are line graphs showing the percentage of MeWo (Figures 3B-3G) or WM115 (Figures 3H-3M) cells labeled with Annexin V after exposure to various concentrations of exemplary compounds. [Figure 3E-G] Figure 3A is a summary table showing EC50 values ​​for Annexin V induction for certain compounds of the invention in MeWo and WM115 cells, and Figures 3B-3M are line graphs showing the percentage of MeWo (Figures 3B-3G) or WM115 (Figures 3H-3M) cells labeled with Annexin V after exposure to various concentrations of exemplary compounds. [Figure 3H-J] Figure 3A is a summary table showing EC50 values ​​for Annexin V induction for certain compounds of the invention in MeWo and WM115 cells, and Figures 3B-3M are line graphs showing the percentage of MeWo (Figures 3B-3G) or WM115 (Figures 3H-3M) cells labeled with Annexin V after exposure to various concentrations of exemplary compounds. [Figure 3K-M] Figure 3A is a summary table showing EC50 values ​​for Annexin V induction for certain compounds of the invention in MeWo and WM115 cells, and Figures 3B-3M are line graphs showing the percentage of MeWo (Figures 3B-3G) or WM115 (Figures 3H-3M) cells labeled with Annexin V after exposure to various concentrations of exemplary compounds. [Figure 4A] Figures 4A-4D are tables showing relative annexin V levels (%) in MeWo and WM115 cells after 6, 24, 48, and 72 hours of exposure to various concentrations of exemplary compounds. Figures 4E-4J are histograms showing the results from Figures 4A-4D. Figures 4K and 4L are histograms showing the percentage of MeWo (Figure 4K) and WM115 (Figure 4L) cells labeled with annexin V after 6 hours of exposure to the indicated concentrations of exemplary compounds. [Figure 4B] Figures 4A-4D are tables showing relative annexin V levels (%) in MeWo and WM115 cells after 6, 24, 48, and 72 hours of exposure to various concentrations of exemplary compounds. Figures 4E-4J are histograms showing the results from Figures 4A-4D. Figures 4K and 4L are histograms showing the percentage of MeWo (Figure 4K) and WM115 (Figure 4L) cells labeled with annexin V after 6 hours of exposure to the indicated concentrations of exemplary compounds. [Figure 4C]Figures 4A-4D are tables showing relative annexin V levels (%) in MeWo and WM115 cells after 6, 24, 48, and 72 hours of exposure to various concentrations of exemplary compounds. Figures 4E-4J are histograms showing the results from Figures 4A-4D. Figures 4K and 4L are histograms showing the percentage of MeWo (Figure 4K) and WM115 (Figure 4L) cells labeled with annexin V after 6 hours of exposure to the indicated concentrations of exemplary compounds. [Figure 4D] Figures 4A-4D are tables showing relative annexin V levels (%) in MeWo and WM115 cells after 6, 24, 48, and 72 hours of exposure to various concentrations of exemplary compounds. Figures 4E-4J are histograms showing the results from Figures 4A-4D. Figures 4K and 4L are histograms showing the percentage of MeWo (Figure 4K) and WM115 (Figure 4L) cells labeled with annexin V after 6 hours of exposure to the indicated concentrations of exemplary compounds. [Figure 4E-G] Figures 4A-4D are tables showing relative annexin V levels (%) in MeWo and WM115 cells after 6, 24, 48, and 72 hours of exposure to various concentrations of exemplary compounds. Figures 4E-4J are histograms showing the results from Figures 4A-4D. Figures 4K and 4L are histograms showing the percentage of MeWo (Figure 4K) and WM115 (Figure 4L) cells labeled with annexin V after 6 hours of exposure to the indicated concentrations of exemplary compounds. [Figure 4H-J] Figures 4A-4D are tables showing relative annexin V levels (%) in MeWo and WM115 cells after 6, 24, 48, and 72 hours of exposure to various concentrations of exemplary compounds. Figures 4E-4J are histograms showing the results from Figures 4A-4D. Figures 4K and 4L are histograms showing the percentage of MeWo (Figure 4K) and WM115 (Figure 4L) cells labeled with annexin V after 6 hours of exposure to the indicated concentrations of exemplary compounds. [Figure 4K]Figures 4A-4D are tables showing relative annexin V levels (%) in MeWo and WM115 cells after 6, 24, 48, and 72 hours of exposure to various concentrations of exemplary compounds. Figures 4E-4J are histograms showing the results from Figures 4A-4D. Figures 4K and 4L are histograms showing the percentage of MeWo (Figure 4K) and WM115 (Figure 4L) cells labeled with annexin V after 6 hours of exposure to the indicated concentrations of exemplary compounds. [Figure 4L] Figures 4A-4D are tables showing relative annexin V levels (%) in MeWo and WM115 cells after 6, 24, 48, and 72 hours of exposure to various concentrations of exemplary compounds. Figures 4E-4J are histograms showing the results from Figures 4A-4D. Figures 4K and 4L are histograms showing the percentage of MeWo (Figure 4K) and WM115 (Figure 4L) cells labeled with annexin V after 6 hours of exposure to the indicated concentrations of exemplary compounds. [Diagram 5] 5A-5K are photomicrographs showing MeWo cell morphology after treatment with various concentrations of CV-8684, CV-8685, CV-8688, DMSO, and staurosporine for 6 hours. [Figure 6] 6A-6K are photomicrographs showing MeWo cell morphology after treatment with various concentrations of CV-8684, CV-8685, CV-8688, DMSO, and staurosporine for 24 hours. [Figure 7] 7A-7K are photomicrographs showing MeWo cell morphology after treatment with various concentrations of CV-8684, CV-8685, CV-8688, DMSO, and staurosporine for 48 hours. [Figure 8] 8A-8K are photomicrographs showing MeWo cell morphology after treatment with various concentrations of CV-8684, CV-8685, CV-8688, DMSO, and staurosporine for 72 hours. [Figure 9] 9A-9K are photomicrographs showing WM115 cell morphology following treatment with various concentrations of CV-8684, CV-8685, CV-8688, DMSO, and staurosporine for 6 hours. [Figure 10]10A-10K are photomicrographs showing WM115 cell morphology following treatment with various concentrations of CV-8684, CV-8685, CV-8688, DMSO, and staurosporine for 24 hours. [Figure 11] 11A-11K are photomicrographs showing WM115 cell morphology following treatment with various concentrations of CV-8684, CV-8685, CV-8688, DMSO, and staurosporine for 48 hours. [Figure 12] 12A-12K are photomicrographs showing WM115 cell morphology following treatment with various concentrations of CV-8684, CV-8685, CV-8688, DMSO, and staurosporine for 72 hours. [Figure 13A] 13A-13D are tables showing the percentage of MeWo and WM115 viable cells remaining after treatment with various concentrations of CV-8684 (FIG. 13A), CV-8685 (FIG. 13B), CV-8688 (FIG. 13C), or staurosporine (FIG. 13D) for 6, 24, 48, and 72 hours. Cell viability was assayed using CellTiter-Glo®. FIGs. 13E-13J are histograms showing the results from FIGs. 13A-13D. FIG. 13K is a summary table comparing the percentage of MeWo and WM115 viable cells after exposure to exemplary concentrations of malassezin, indolocarbazole, compound II, and staurosporine for 24, 48, and 72 hours. [Figure 13B] 13A-13D are tables showing the percentage of MeWo and WM115 viable cells remaining after treatment with various concentrations of CV-8684 (FIG. 13A), CV-8685 (FIG. 13B), CV-8688 (FIG. 13C), or staurosporine (FIG. 13D) for 6, 24, 48, and 72 hours. Cell viability was assayed using CellTiter-Glo®. FIGs. 13E-13J are histograms showing the results from FIGs. 13A-13D. FIG. 13K is a summary table comparing the percentage of MeWo and WM115 viable cells after exposure to exemplary concentrations of malassezin, indolocarbazole, compound II, and staurosporine for 24, 48, and 72 hours. [Figure 13C]13A-13D are tables showing the percentage of MeWo and WM115 viable cells remaining after treatment with various concentrations of CV-8684 (FIG. 13A), CV-8685 (FIG. 13B), CV-8688 (FIG. 13C), or staurosporine (FIG. 13D) for 6, 24, 48, and 72 hours. Cell viability was assayed using CellTiter-Glo®. FIGs. 13E-13J are histograms showing the results from FIGs. 13A-13D. FIG. 13K is a summary table comparing the percentage of MeWo and WM115 viable cells after exposure to exemplary concentrations of malassezin, indolocarbazole, compound II, and staurosporine for 24, 48, and 72 hours. [Figure 13D] 13A-13D are tables showing the percentage of MeWo and WM115 viable cells remaining after treatment with various concentrations of CV-8684 (FIG. 13A), CV-8685 (FIG. 13B), CV-8688 (FIG. 13C), or staurosporine (FIG. 13D) for 6, 24, 48, and 72 hours. Cell viability was assayed using CellTiter-Glo®. FIGs. 13E-13J are histograms showing the results from FIGs. 13A-13D. FIG. 13K is a summary table comparing the percentage of MeWo and WM115 viable cells after exposure to exemplary concentrations of malassezin, indolocarbazole, compound II, and staurosporine for 24, 48, and 72 hours. [Fig. 13E-G] 13A-13D are tables showing the percentage of MeWo and WM115 viable cells remaining after treatment with various concentrations of CV-8684 (FIG. 13A), CV-8685 (FIG. 13B), CV-8688 (FIG. 13C), or staurosporine (FIG. 13D) for 6, 24, 48, and 72 hours. Cell viability was assayed using CellTiter-Glo®. FIGs. 13E-13J are histograms showing the results from FIGs. 13A-13D. FIG. 13K is a summary table comparing the percentage of MeWo and WM115 viable cells after exposure to exemplary concentrations of malassezin, indolocarbazole, compound II, and staurosporine for 24, 48, and 72 hours. [Figure 13H-J]13A-13D are tables showing the percentage of MeWo and WM115 viable cells remaining after treatment with various concentrations of CV-8684 (FIG. 13A), CV-8685 (FIG. 13B), CV-8688 (FIG. 13C), or staurosporine (FIG. 13D) for 6, 24, 48, and 72 hours. Cell viability was assayed using CellTiter-Glo®. FIGs. 13E-13J are histograms showing the results from FIGs. 13A-13D. FIG. 13K is a summary table comparing the percentage of MeWo and WM115 viable cells after exposure to exemplary concentrations of malassezin, indolocarbazole, compound II, and staurosporine for 24, 48, and 72 hours. [Figure 13K] 13A-13D are tables showing the percentage of MeWo and WM115 viable cells remaining after treatment with various concentrations of CV-8684 (FIG. 13A), CV-8685 (FIG. 13B), CV-8688 (FIG. 13C), or staurosporine (FIG. 13D) for 6, 24, 48, and 72 hours. Cell viability was assayed using CellTiter-Glo®. FIGs. 13E-13J are histograms showing the results from FIGs. 13A-13D. FIG. 13K is a summary table comparing the percentage of MeWo and WM115 viable cells after exposure to exemplary concentrations of malassezin, indolocarbazole, compound II, and staurosporine for 24, 48, and 72 hours. [Figure 14A] 14A-14D are tables showing lactate dehydrogenase ("LDH") release levels from MeWo and WM115 cells after treatment with various concentrations of CV-8684 (FIG. 14A), CV-8685 (FIG. 14B), CV-8688 (FIG. 14C), or staurosporine (FIG. 14D) for 6, 24, 48, and 72 hours. FIGs. 14E-14J are histograms showing the results from FIGs. 14A-14D. FIGs. 14K and 14L are histograms showing lactate dehydrogenase levels after exposure of MeWo (FIG. 14K) and WM115 (FIG. 14L) cells to exemplary concentrations of malassezin, carbazole, compound II, and staurosporine for 24 hours. [Figure 14B]14A-14D are tables showing lactate dehydrogenase ("LDH") release levels from MeWo and WM115 cells after treatment with various concentrations of CV-8684 (FIG. 14A), CV-8685 (FIG. 14B), CV-8688 (FIG. 14C), or staurosporine (FIG. 14D) for 6, 24, 48, and 72 hours. FIGs. 14E-14J are histograms showing the results from FIGs. 14A-14D. FIGs. 14K and 14L are histograms showing lactate dehydrogenase levels after exposure of MeWo (FIG. 14K) and WM115 (FIG. 14L) cells to exemplary concentrations of malassezin, carbazole, compound II, and staurosporine for 24 hours. [Figure 14C] 14A-14D are tables showing lactate dehydrogenase ("LDH") release levels from MeWo and WM115 cells after treatment with various concentrations of CV-8684 (FIG. 14A), CV-8685 (FIG. 14B), CV-8688 (FIG. 14C), or staurosporine (FIG. 14D) for 6, 24, 48, and 72 hours. FIGs. 14E-14J are histograms showing the results from FIGs. 14A-14D. FIGs. 14K and 14L are histograms showing lactate dehydrogenase levels after exposure of MeWo (FIG. 14K) and WM115 (FIG. 14L) cells to exemplary concentrations of malassezin, carbazole, compound II, and staurosporine for 24 hours. [Figure 14D] 14A-14D are tables showing lactate dehydrogenase ("LDH") release levels from MeWo and WM115 cells after treatment with various concentrations of CV-8684 (FIG. 14A), CV-8685 (FIG. 14B), CV-8688 (FIG. 14C), or staurosporine (FIG. 14D) for 6, 24, 48, and 72 hours. FIGs. 14E-14J are histograms showing the results from FIGs. 14A-14D. FIGs. 14K and 14L are histograms showing lactate dehydrogenase levels after exposure of MeWo (FIG. 14K) and WM115 (FIG. 14L) cells to exemplary concentrations of malassezin, carbazole, compound II, and staurosporine for 24 hours. [Fig. 14E-G]14A-14D are tables showing lactate dehydrogenase ("LDH") release levels from MeWo and WM115 cells after treatment with various concentrations of CV-8684 (FIG. 14A), CV-8685 (FIG. 14B), CV-8688 (FIG. 14C), or staurosporine (FIG. 14D) for 6, 24, 48, and 72 hours. FIGs. 14E-14J are histograms showing the results from FIGs. 14A-14D. FIGs. 14K and 14L are histograms showing lactate dehydrogenase levels after exposure of MeWo (FIG. 14K) and WM115 (FIG. 14L) cells to exemplary concentrations of malassezin, carbazole, compound II, and staurosporine for 24 hours. [Fig. 14H-J] 14A-14D are tables showing lactate dehydrogenase ("LDH") release levels from MeWo and WM115 cells after treatment with various concentrations of CV-8684 (FIG. 14A), CV-8685 (FIG. 14B), CV-8688 (FIG. 14C), or staurosporine (FIG. 14D) for 6, 24, 48, and 72 hours. FIGs. 14E-14J are histograms showing the results from FIGs. 14A-14D. FIGs. 14K and 14L are histograms showing lactate dehydrogenase levels after exposure of MeWo (FIG. 14K) and WM115 (FIG. 14L) cells to exemplary concentrations of malassezin, carbazole, compound II, and staurosporine for 24 hours. [Figure 14K] 14A-14D are tables showing lactate dehydrogenase ("LDH") release levels from MeWo and WM115 cells after treatment with various concentrations of CV-8684 (FIG. 14A), CV-8685 (FIG. 14B), CV-8688 (FIG. 14C), or staurosporine (FIG. 14D) for 6, 24, 48, and 72 hours. FIGs. 14E-14J are histograms showing the results from FIGs. 14A-14D. FIGs. 14K and 14L are histograms showing lactate dehydrogenase levels after exposure of MeWo (FIG. 14K) and WM115 (FIG. 14L) cells to exemplary concentrations of malassezin, carbazole, compound II, and staurosporine for 24 hours. [Figure 14L]14A-14D are tables showing lactate dehydrogenase ("LDH") release levels from MeWo and WM115 cells after treatment with various concentrations of CV-8684 (FIG. 14A), CV-8685 (FIG. 14B), CV-8688 (FIG. 14C), or staurosporine (FIG. 14D) for 6, 24, 48, and 72 hours. FIGs. 14E-14J are histograms showing the results from FIGs. 14A-14D. FIGs. 14K and 14L are histograms showing lactate dehydrogenase levels after exposure of MeWo (FIG. 14K) and WM115 (FIG. 14L) cells to exemplary concentrations of malassezin, carbazole, compound II, and staurosporine for 24 hours. [Figure 15A] FIG. 1 shows raw data and line graphs of aryl hydrocarbon receptor ("AhR") activation in HepG2 cells stably transfected with an AhR-responsive luciferase reporter gene plasmid upon exposure to various concentrations of omeprazole. [Figure 15B] FIG. 1 shows raw data and a line graph of aryl hydrocarbon receptor ("AhR") activation in HepG2 cells stably transfected with an AhR-responsive luciferase reporter gene plasmid upon exposure to various concentrations of CV-8684. [Figure 15C] FIG. 1 shows raw data and a line graph of aryl hydrocarbon receptor ("AhR") activation in HepG2 cells stably transfected with an AhR-responsive luciferase reporter gene plasmid upon exposure to various concentrations of CV-8685. [Figure 15D] FIG. 1 shows raw data and a line graph of aryl hydrocarbon receptor ("AhR") activation in HepG2 cells stably transfected with an AhR-responsive luciferase reporter gene plasmid upon exposure to various concentrations of CV-8686. [Figure 15E] FIG. 1 shows raw data and line graphs of aryl hydrocarbon receptor ("AhR") activation in HepG2 cells stably transfected with an AhR-responsive luciferase reporter gene plasmid upon exposure to various concentrations of CV-8688. [Figure 15F] The EC50 value for each test compound is shown. [Figure 16A-E] 16A-16K are photographs of MelanoDerm™ matrix either 0 or 7 days after exposure to untreated (FIG. 16A), sterile deionized water (FIG. 16B), 1% kojic acid (FIG. 16C), 0.2% DMSO (FIG. 16D), 0.05% DMSO (FIG. 16E), 200 μM CV-8684 (FIG. 16F), 50 μM CV-8684 (FIG. 16G), 200 μM CV-8686 (FIG. 16H), 50 μM CV-8686 (FIG. 16I), 200 μM CV-8688 (FIG. 16J), and 50 μM CV-8688 (FIG. 16K). [Fig. 16F-K] 16A-16K are photographs of MelanoDerm™ matrix either 0 or 7 days after exposure to untreated (FIG. 16A), sterile deionized water (FIG. 16B), 1% kojic acid (FIG. 16C), 0.2% DMSO (FIG. 16D), 0.05% DMSO (FIG. 16E), 200 μM CV-8684 (FIG. 16F), 50 μM CV-8684 (FIG. 16G), 200 μM CV-8686 (FIG. 16H), 50 μM CV-8686 (FIG. 16I), 200 μM CV-8688 (FIG. 16J), and 50 μM CV-8688 (FIG. 16K). [Figure 17A-E] 17A-17K are 15x photomicrographs of MelanoDerm™ Matrix either 0 or 7 days after exposure to untreated (FIG. 17A), sterile deionized water (FIG. 17B), 1% kojic acid (FIG. 17C), 0.2% DMSO (FIG. 17D), 0.05% DMSO (FIG. 17E), 200 μM CV-8684 (FIG. 17F), 50 μM CV-8684 (FIG. 17G), 200 μM CV-8686 (FIG. 17H), 50 μM CV-8686 (FIG. 17I), 200 μM CV-8688 (FIG. 17J), and 50 μM CV-8688 (FIG. 17K). [Fig. 17F-K]17A-17K are 15x photomicrographs of MelanoDerm™ Matrix either 0 or 7 days after exposure to untreated (FIG. 17A), sterile deionized water (FIG. 17B), 1% kojic acid (FIG. 17C), 0.2% DMSO (FIG. 17D), 0.05% DMSO (FIG. 17E), 200 μM CV-8684 (FIG. 17F), 50 μM CV-8684 (FIG. 17G), 200 μM CV-8686 (FIG. 17H), 50 μM CV-8686 (FIG. 17I), 200 μM CV-8688 (FIG. 17J), and 50 μM CV-8688 (FIG. 17K). [Figure 18] 18A-18F are photographs of zebrafish exposed to no treatment (FIG. 18A), DMSO (FIG. 18B), phenylthiourea ("PTU") (FIG. 18C), and 2.5 μM Compound II (FIG. 18D), 5 μM Compound II (FIG. 18E), and 10 μM Compound II (FIG. 18F). Red arrows indicate normal melanocytes. [Figure 19] 19A-19F are photographs of zebrafish exposed to untreated (FIG. 19A), DMSO (FIG. 19B), phenylthiourea ("PTU") (FIG. 19C), and 0.3 μM Compound II (FIG. 19D), 1 μM Compound II (FIG. 19E), and 3 μM Compound II (FIG. 19F). Red arrows indicate normal melanocytes. Yellow arrows indicate abnormally small melanocytes. [Figure 20] 1 is a summary table showing the number and percentage (%) of zebrafish that exhibited reduced skin pigmentation after exposure to exemplary conditions. The last six columns show the effect of various concentrations of Compound II. [Figure 21] 21A-21E are photographs of zebrafish that were untreated (FIG. 21A), treated with DMSO (FIG. 21B), PTU (FIG. 21C), 0.5 μM (FIG. 21D), and 1.5 μM (FIG. 21E). The lower panels contain areas with inverted color schemes. [Figure 22A] 21A-21E are histograms showing pigmentation density as measured by pigmented pixels / mm3 from photographs of zebrafish embryos illustrated in FIGS. 21A-21E. [Figure 22B]21A-21E are histograms showing total pixel counts from the photographs of zebrafish embryos illustrated in FIGS. 21A-21E. [Figure 23A] 1 is a mass spectrum of CV-8684 in DMSO. [Figure 23B] 1 is a mass spectrum of CV-8684 in RPMI medium. [Figure 23C] 1 is a mass spectrum of CV-8684 in DMEM. [Figure 23D] 1 is a mass spectrum of CV-8686 in DMSO. [Figure 23E] 1 is a mass spectrum of CV-8686 in RPMI medium. [Figure 23F] 1 is a mass spectrum of CV-8686 in DMEM. [Figure 23G] 1 is a mass spectrum of CV-8688 in DMSO. [Figure 23H] 1 is a mass spectrum of CV-868 in RPMI medium. [Figure 23I] 1 is a mass spectrum of CV-8688 in DMEM. [Figure 23J] 1 is a summary table showing the percentage of test compound remaining in the exemplary medium after 2 hours of incubation. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0044] One embodiment of the present invention is a compound for skin whitening, which is a chemical analogue of the compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0045] As used herein, the term "compound" refers to two or more atoms linked by one or more chemical bonds. In the present invention, chemical bonds include, but are not limited to, covalent bonds, ionic bonds, hydrogen bonds, and van der Waals interactions. Covalent bonds in the present invention include single bonds, double bonds, and triple bonds. Compounds of the present invention include, but are not limited to, organic molecules.

[0046] The organic compounds / molecules of the present invention include linear, branched, and cyclic hydrocarbons with or without functional groups. x~y The term "alkyl," when used with a chemical moiety such as alkyl, alkenyl, alkynyl, or alkoxy, is meant to include groups containing x to y carbons in the chain. For example, "C x~y The term "alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups including straight-chain and branched-chain alkyl groups containing x to y carbons in the chain, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl. x~y alkenyl" and "C x~y The term "alkynyl" refers to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but which contain at least one double or triple bond respectively.

[0047] The term "aliphatic," as used herein, means a group composed of carbon and hydrogen atoms that does not contain an aromatic ring. Thus, aliphatic groups include alkyl, alkenyl, alkynyl, and carbocyclyl groups.

[0048] The term "alkyl" refers to groups that are saturated aliphatic groups not having a ring structure, including straight-chain and branched-chain alkyl groups.

[0049] The term "alkenyl," as used herein, means an aliphatic group containing at least one double bond.

[0050] The term "alkynyl," as used herein, refers to an aliphatic group containing at least one triple bond.

[0051] As used herein, an "aromatic compound", "aromatic", or compound containing an "aromatic ring" is an aryl or heteroaryl compound. The term "aryl" as used herein includes substituted or unsubstituted monocyclic aromatic groups in which each atom of the ring is carbon. Preferably, the ring is 3-8 membered, more preferably 6 membered. 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 which is aromatic, e.g., the other cyclic ring may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like. The term "heteroaryl" includes substituted or unsubstituted aromatic monocyclic structures, preferably 3-8 membered rings, more preferably 5-7 membered rings, and even more preferably 5-6 membered rings, containing at least one heteroatom, preferably 1-4 heteroatoms, more preferably 1-2 heteroatoms. The term "heteroaryl" 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 which is heteroaromatic, e.g., the other cyclic ring may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heteroaryl groups include, for example, pyrrole, furan, thiophene, indole, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine. Preferably, certain compounds of the invention contain at least one, preferably two, indole groups, and at least one aldehyde group.

[0052] The term "substituted" means a moiety having at least one substituent replacing a hydrogen atom on one or more carbons of the backbone. It is understood that "substituted" or "substituted with" includes the implicit proviso that such substitution is subject to the permissible valences of the substituted atom and substituent, as well as that the substitution results in a stable compound, e.g., it does not spontaneously result in transformations such as rearrangement, cyclization, elimination, and the like. Permissible substituents can be one or more and the same or different for appropriate organic compounds.

[0053] As used herein, "skin whitening" and grammatical variations thereof generally refer to the reduction of all skin pigmentation, actual or perceived. Skin whitening methods have been used to reduce hyperpigmentation in hyperpigmented areas of the skin resulting from age, sun exposure, or hyperpigmentation disorders. The compounds and compositions of the present invention, when applied, for example, to the skin of a subject, can reduce pigmentation, resulting in the skin appearing lighter or whiter than before said application. Skin pigmentation can be assessed in a number of ways, including, for example, the von Luschan color scale (von Luschan These include, but are not limited to, visual assessment using the chromatic scale, the Fitzpatrick skin classification test (Fitzpatrick et al., 1988), and the Taylor Hyperpigmentation Scale (Taylor et al., 2005), as well as reflectance spectrophotometry (Zonios, et al., 2001). For example, the Fitzpatrick skin classification test includes six skin types (I-VI), and as the term "whitened" is used herein, type VI skin that falls below type V is "whitened". As discussed further below, skin whitening can occur through a number of phenomena. These phenomena include, but are not limited to, modulation of melanocyte activity, induction of melanocyte apoptosis, agonism of the aryl hydrocarbon receptor (AhR), or modulation of melanin production, melanosome biogenesis, or melanosome transfer.

[0054] Certain compounds of the invention are produced by, isolated from, or isolatable from Malassezia yeasts. Malassezia yeasts are yeasts of the genus Malassezia, which include, but are not limited to, Malassezia globosa, Malassezia restricta, Malassezia furfur, Malassezia sympodialis, Malassezia slooffiae, Malassezia obtusa, Malassezia pachydermatis, Malassezia dermatis, Malassezia Malassezia yeasts include, but are not limited to, Malassezia japonica, Malassezia nana, Malassezia yamatoensis, Malassezia equine, Malassezia caprae, and Malassezia cuniculi. (Gueho, et al., 1996; Gaitanis, et al., 2013). Malassezia yeasts are part of the normal human skin flora and typically do not produce pathogenic effects. However, Malassezia yeasts can cause a number of diseases, including, but not limited to, tinea versicolor (both tinea nigricans and tinea alba varieties), seborrheic dermatitis, dandruff, atopic dermatitis, Malassezia folliculitis, psoriasis, and confluent reticular papillomatosis. (Gaitanis, et al., 2013).

[0055] As used herein, the term "chemical analog" refers to a compound that is structurally related to a parent compound and contains different functional groups or substituents. For example, the parent compound of the present invention is malassezin, and a chemical analog of malassezin contains certain functional groups and substituents that are different from malassezin. The chemical analogs of the present invention can have significant advantages over a given parent compound, including a pharmacokinetic profile suitable for use as a cosmetic. In some embodiments, the chemical analogs are produced from the parent molecule by one or more chemical reactions. In other embodiments, the chemical analogs of the present invention can be produced using alternative synthetic schemes that are not derived from the parent compound.

[0056] A compound of the invention is "produced by a Malassezia yeast" if the yeast synthesizes, secretes, accumulates, or otherwise produces the compound of the invention during its life cycle under the proper growth conditions. Malassezia yeast secretes different compounds depending on what is supplemented in its growth medium. (Nazzaro-Porro, et al., 1978). The invention includes any compound produced by a Malassezia yeast under any growth conditions, although preferred compounds include, for example, malassedins and their chemical analogs.

[0057] In one aspect of this embodiment, the compound produced by the Malassezia yeast has the structure of Formula (I): [ka]

[0058] In another aspect of this embodiment, the compound is a chemical analogue of malassezin.

[0059] Malassedin is an example of a compound produced by the Malassezia yeast of the present invention. Malassedin, also known as 2-(1H-indol-3-ylmethyl)-1H-indole-3-carbaldehyde, is a tryptophan metabolite originally isolated from Malassezia furfur. Malassedin is a known agonist of the aryl hydrocarbon receptor (AhR), a receptor involved in cell proliferation, differentiation, and gene expression. (Wille et al., 2001). Malassedin also induces apoptosis in primary human melanocytes. (Kramer, et al., 2005). Recently, certain chemical analogs of malassedin have been synthesized by Winston-McPherson and coworkers, who have investigated the AhR agonist activity of the analogs. (Winston-McPherson, et al., 2014).

[0060] Another embodiment of the present invention is a compound for inducing apoptosis of melanocytes, the compound being a chemical analogue of the compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0061] As used herein, the term "melanocyte" generally refers to dendritic cells of the epidermis that synthesize tyrosinase and synthesize the pigment melanin within melanosomes. The melanocytes of the present invention exhibit upregulation of certain genes. These genes include, but are not limited to, the following: tyrosinase (oculocutaneous albinism IA), microphthalmia-associated transcription factor, α-2-macroglobulin, tyrosinase-related protein 1, solute carrier family 16, GS3955 protein, v-kit Hardy-Zuckerman4 feline sarcoma virus, oculocutaneous albinism II, deleted in esophageal cancer 1, melan-A, SRY-box 10, ATPase, class V, type 10C, matrix metalloprotease 1, latent transforming growth factor βb, ATP-binding cassette, subfamily C, hydroxysteroid dehydrogenase 15, transmembrane 7 superfamily member 1, glutaminylpeptide cyclotransferase, and one or more of the other genes identified by Lee and co-workers (Lee, et al., 2013).

[0062] Melanocytes, like many other cell types, undergo programmed cell death, i.e., apoptosis. The apoptotic pathway of melanocytes is known to those skilled in the art (Wang, et al., 2014), and the apoptotic pathway has been generally reviewed by Elmore (Elmore, 2007). The compounds or compositions of the present invention "induce" apoptosis of melanocytes, for example, by causing activation of certain apoptosis-promoting signaling pathways or causing inhibition of certain anti-apoptotic pathways in melanocytes. It is envisioned that the compounds or compositions of the present invention can directly activate / inhibit pathways related to apoptosis by directly interacting with the signaling molecules of the pathway or indirectly interacting with the molecules of the pathway through direct interaction with one or more intermediate molecules that do not typically function within the pathway.

[0063] In one aspect of this embodiment, the compound produced by the Malassezia yeast has the structure of Formula (I): [ka]

[0064] In another aspect of this embodiment, the compound is a chemical analogue of malassezin.

[0065] A further embodiment of the present invention is a compound for modulating melanocyte activity, which is a chemical analogue of the compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0066] Melanocyte activity can be modulated in a number of ways contemplated in the present invention, including, but not limited to, inducing apoptosis of melanocytes or altering melanocyte gene expression, cell motility, cell proliferation, melanin production, melanosome biogenesis, or melanosome transport.

[0067] In one aspect of this embodiment, the compound produced by the Malassezia yeast has the structure of Formula (I): [ka]

[0068] In another aspect of this embodiment, the compound is a chemical analogue of malassegin.

[0069] An additional embodiment of the present invention is a compound for agonizing the aryl hydrocarbon receptor (AhR), which is a chemical analog of a compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0070] As used herein, the terms "agonist," "agonize," and grammatical variations thereof refer to a molecule that triggers (e.g., initiates or promotes), partially or fully enhances, stimulates, or activates one or more biological activities. Agonists of the present invention include naturally occurring as well as synthetic substances.

[0071] The aryl hydrocarbon receptor (AhR) of the present invention is any aryl hydrocarbon receptor naturally occurring in a subject as described herein. Aryl hydrocarbon receptors are known to those skilled in the art. (Noakes, 2015). Agonists of the aryl hydrocarbon receptor include tryptophan-related compounds such as kynurenine, kynurenic acid, cinnabarinic acid, and 6-formylindolo[3,2-b]carbazole (FICZ), among others. Not limited thereto. Malassezin is also known as an agonist of the aryl hydrocarbon receptor. (Wille, et al., 2001).

[0072] In one aspect of this embodiment, the compound produced by the Malassezia yeast has the structure of Formula (I): [ka]

[0073] In another aspect of this embodiment, the compound is a chemical analogue of malassegin.

[0074] Another embodiment of the present invention is a compound for improving hyperpigmentation caused by hyperpigmentation disorders, the compound being a chemical analogue of the compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0075] As used herein, the compounds, compositions and methods of the present invention can be used to improve the hyperpigmentation caused by hyperpigmentation disorder, for example, by reducing the level of hyperpigmentation in the area affected by hyperpigmentation disorder, delaying further hyperpigmentation, or preventing further hyperpigmentation from occurring.However, improving the hyperpigmentation caused by hyperpigmentation disorder does not require that the desired physiological response or outcome is achieved in each and every subject or subject group, since not every subject may respond to a particular dosing protocol, regimen, or process.Therefore, a given subject or subject group may not respond or respond inappropriately to dosing, while other subjects or subject groups may respond, and thus feel the improvement of their hyperpigmentation disorder.

[0076] As used herein, the term "hyperpigmentation" refers to a real or perceived excessively dark skin disorder. This skin disorder may be realized, for example, due to age, excessive sun exposure, or a disease or condition that results in dark skin areas. The dark skin areas may be in the form of dark spots, birthmarks, or relatively large areas. This skin disorder may be, for example, due to the perception by an individual that their skin tone is perceived as being too dark. An individual may have a cosmetic desire to lighten their skin tone.

[0077] Hyperpigmentation disorders include not only disorders in which hyperpigmentation is a primary symptom, but also disorders in which hyperpigmentation occurs as a secondary symptom. The hyperpigmentation disorders of the present invention include, but are not limited to, congenital hyperpigmentation disorders and acquired hyperpigmentation disorders. The congenital hyperpigmentation disorders of the present invention include epidermal hyperpigmentation (nevus cell nevus, Spitz nevus, and flat nevus), dermal hyperpigmentation (blue nevus, nevus of Ota, dermal melanocytosis, nevus of Ito, and Mongolian spot), freckles, reticulate acropigmentation of Kitamura, acral pigmentation / acral lentiginosis, and multiple lentigines syndrome (generalized lentigines syndrome, Leopard syndrome, inherited patterned lentiginosis, Carney complex, Peutz-Jeghers syndrome, Laugier-Hunziker-Baran syndrome, and Cronkhite Acquired hyperpigmentation disorders of the present invention include, but are not limited to, those involving senile lentigo / lentigines, melasma / chloasma, Lille melanosis, melanoma of the lips, penile / vulvar melanosis, facial follicular erythematous melanosis (Kitamura), UV-induced hyperpigmentation (sunburn and actinic petaloid pigmentation), post-inflammatory hyperpigmentation (frictional melanosis and dyschromatosis perstans), chemical / drug-induced hyperpigmentation (polychlorinated biphenyls, arsenic, 5-FU, bleomycin, cyclophosphamide, methotrexate, chlorpromazine, phenytoin, tetracycline, and chloroquine), pigment demarcation striae, and foreign body deposits (such as carotene, silver, gold, mercury, bismuth, and tattoos). Hyperpigmentation associated with systemic disorders includes metabolic / enzymatic disorders (hemochromatosis, Wilson's disease, Gaucher's disease, Niemann-Pick disease, amyloidosis, histochromia, acanthosis nigricans, and porphyria cutanea tarda), endocrine disorders (Addison's disease, Cushing's syndrome, and hyperthyroidism), nutritional disorders (pellagra, vitamin B12 deficiency, folate deficiency, vagrant disease, and prurigo pigmentosa), mastocytosis, collagen diseases, hepatic dysfunction, and renal dysfunction. Hyperpigmentation can also be associated with infectious diseases (measles, syphilis, and Malassezia furfur) and syndromes (von Recklinghausen's disease, Sotos syndrome, POEMS syndrome, Naegeli syndrome, Cantu syndrome, McCune-Albright syndrome, Watson syndrome, and Bloom syndrome) (Yamaguchi, et al., 2014).

[0078] In one aspect of this embodiment, the compound produced by the Malassezia yeast has the structure of Formula (I): [ka]

[0079] In another aspect of this embodiment, the compound is a chemical analogue of malassezin.

[0080] A further embodiment of the present invention is a compound for modulating melanin production, which is a chemical analogue of the compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0081] Melanin is a naturally occurring pigment that gives skin and hair its color. A schematic diagram of skin is shown in FIG. 1A. Melanin is produced by melanocytes in organelles known as melanosomes. The compounds or compositions of the present invention modulate melanin production in a subject, for example, by modulating melanosome biogenesis and directly or indirectly inhibiting melanin synthesis at the enzyme level.

[0082] Melanosome biogenesis occurs in four stages. Stage I is characterized by premelanosomes, which are essentially non-pigmented vacuoles. In stage II, the premelanosomes give rise to striations where melanin is deposited in stage III. Stage IV results in mature melanosomes that are rich in melanin content. The compounds and compositions of the present invention modulate melanosome biogenesis by inhibiting or attenuating the biological processes that normally drive any or all of these stages. (Wasmeier, et al., 2008).

[0083] Melanin synthesis primarily requires three enzymes: tyrosinase, tyrosinase-related protein 1, and dopachrome isomerase. Additional factors that affect the intracellular trafficking of these enzymes include, but are not limited to, BLOC-1, OA1, and SLC45A2. The compounds and compositions of the present invention can modulate melanin production, for example, by inhibiting or attenuating the activity of any of these enzymes or factors. (Yamaguchi, et al., 2014).

[0084] Once melanosomes are formed and melanin is synthesized, they must be transferred from epidermal melanocytes to skin and hair keratinocytes. Melanosomes originate near the nucleus of melanocytes and are transported along microtubules and actin fibers to the periphery of the melanocyte. The compounds and compositions of the present invention regulate melanosome transfer by interfering with any of the biological processes that lead to the transport of melanosomes from the perinuclear region to the periphery of the melanocyte and into adjacent keratinocytes. A schematic diagram of melanin synthesis, melanin transport, and melanocyte apoptosis is shown in Figure 1B.

[0085] In one aspect of this embodiment, the compound produced by the Malassezia yeast has the structure of Formula (I): [ka]

[0086] In another aspect of this embodiment, the compound is a chemical analogue of malassezin.

[0087] An additional embodiment of the present invention is a compound for modulating melanosome biogenesis, which is a chemical analogue of the compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0088] In one aspect of this embodiment, the compound produced by the Malassezia yeast has the structure of Formula (I): [ka]

[0089] In another aspect of this embodiment, the compound is a chemical analogue of malassezin.

[0090] Another embodiment of the present invention is a compound for modulating melanosome transport, which is a chemical analogue of the compound produced by Malassezia yeast, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof.

[0091] In one aspect of this embodiment, the compound produced by the Malassezia yeast has the structure of Formula (I): [ka]

[0092] In another aspect of this embodiment, the compound is a chemical analogue of malassezin.

[0093] A further embodiment of the invention is a composition comprising Malassezia yeast and a cosmetically or pharma- ceutically acceptable vehicle, diluent, or carrier.

[0094] An additional embodiment of the invention is a composition comprising a compound isolated or isolatable from Malassezia yeast and a cosmetically or pharma- ceutically acceptable vehicle, diluent, or carrier.

[0095] The compounds isolated from the Malassezia yeast of the present invention are necessarily present in the Malassezia yeast before isolation or are produced by the Malassezia yeast.Therefore, the compounds isolated from the Malassezia yeast are obtained from actual yeast cells.Standard protocols for extracting compounds from cell material are known to those skilled in the art.

[0096] Compounds that are isolable from Malassezia yeast do not necessarily have to be obtained from actual yeast cells. Instead, synthetic reactions can be used to generate compounds that are produced in yeast without the intervention of actual yeast cells. Organic synthetic reactions are well known to those skilled in the art and can be used in this regard.

[0097] Another embodiment of the invention is a composition comprising any of the compounds disclosed herein, including analogs, and a cosmetically or pharma- ceutically acceptable vehicle, diluent, or carrier.

[0098] A further embodiment of the invention is a method of lightening the skin in a subject, comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0099] As used herein, the term "contact" and grammatical variations thereof refer to bringing two or more materials into close enough proximity so that they can interact. Thus, for illustrative purposes only, the compounds of the present invention can contact melanocytes, for example, by interacting with receptors on the surface of the melanocytes. Similarly, the compositions of the present invention can contact a human subject, for example, by being applied directly to the skin of the subject.

[0100] As used herein, "subject" refers to a mammalian cell, tissue, organism, or population thereof. Subjects of the present invention are preferably human, including human cells, human tissue, and humans, but also include primates, farm animals, livestock, laboratory animals, and the like. Some examples of agricultural animals include cows, pigs, horses, goats, and the like. Some livestock animals include Examples of laboratory animals include dogs, cats, etc. Some examples of laboratory animals include primates, rats, mice, rabbits, guinea pigs, etc.

[0101] An additional embodiment of the invention is a method for inducing apoptosis of melanocytes in a subject, comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0102] Another embodiment of the invention is a method for modulating melanocyte activity in a subject, comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0103] A further embodiment of the invention is a method for agonizing the aryl hydrocarbon receptor (AhR), comprising contacting a subject with any of the compounds or compositions disclosed herein.

[0104] An additional embodiment of the invention is a method for ameliorating hyperpigmentation caused by a hyperpigmentation disorder in a subject in need thereof, comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0105] As used herein, a subject "in need" of amelioration of hyperpigmentation caused by a hyperpigmentation disorder includes a subject who actually needs amelioration or a subject who perceives the need for amelioration.

[0106] Another embodiment of the invention is a method for modulating melanin production in a subject, comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0107] A further embodiment of the invention is a method for modulating melanosome biogenesis in a subject, comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0108] An additional embodiment of the invention is a method for modulating melanosome transport in a subject, comprising contacting the subject with any of the compounds or compositions disclosed herein.

[0109] Another embodiment of the present invention is a compound having the formula (II): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl, and at least one of R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 is methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0110] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] and [ka]

[0111] A further embodiment of the present invention is a compound having the formula (III): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl, and at least one of R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 is methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0112] In one aspect of this embodiment, the compound is: [ka]

[0113] An additional embodiment of the present invention is a compound for whitening the skin, the compound having the formula (II): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0114] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] , [ka] and [ka]

[0115] Another embodiment of the present invention is a compound for skin whitening, the compound having the formula (III): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0116] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] and [ka]

[0117] A further embodiment of the present invention is a compound for inducing apoptosis of melanocytes, the compound having formula (II): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0118] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] , [ka] and [ka]

[0119] Further embodiments of the present invention are directed to compounds for inducing apoptosis of melanocytes. The compound has the formula (III): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0120] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] and [ka]

[0121] Another embodiment of the present invention is a compound for agonizing the aryl hydrocarbon receptor (AhR), the compound having the formula (II): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0122] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] , [ka] and [ka]

[0123] A further embodiment of the present invention is a compound for agonizing the aryl hydrocarbon receptor (AhR), the compound having the formula (III): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl), or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof.

[0124] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] and [ka]

[0125] An additional embodiment of the present invention is a composition comprising a compound of formula (II): [ka] (wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl), or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof, and a cosmetically or pharma- ceutically acceptable vehicle, diluent, or carrier.

[0126] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] , [ka] and [ka]

[0127] Another embodiment of the present invention is a composition comprising a compound of formula (III): [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof, and a cosmetically or pharma- ceutically acceptable vehicle, diluent, or carrier.

[0128] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] and [ka]

[0129] A further embodiment of the present invention is a method for whitening the skin in a subject, comprising administering to a subject a compound of formula (II): [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, contacting a subject with a compound having the structure:

[0130] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] , [ka] and [ka]

[0131] An additional embodiment of the present invention is a method for lightening the skin in a subject, comprising: [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, to a subject.

[0132] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] and [ka]

[0133] Another embodiment of the present invention is a method for inducing apoptosis of melanocytes in a subject, the method comprising administering to a subject a compound of formula (II): [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, contacting a subject with a compound having the structure:

[0134] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] , [ka] and [ka]

[0135] A further embodiment of the invention is a method for inducing apoptosis of melanocytes in a subject, comprising administering to a subject a compound of formula (III): [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, to a subject.

[0136] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] and [ka]

[0137] An additional embodiment of the invention is a method for agonizing the aryl hydrocarbon receptor (AhR) in a subject, the method comprising administering to a subject a compound of formula (II): [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, contacting a subject with a compound having the structure:

[0138] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] , [ka] and [ka]

[0139] Another embodiment of the present invention is a method for agonizing the aryl hydrocarbon receptor (AhR) in a subject, comprising administering to a subject a compound of formula (III): [ka] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl, or a crystalline form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, to a subject.

[0140] In one aspect of this embodiment, the compound is selected from the group consisting of: [ka] and [ka]

[0141] As used herein, the term "composition" refers not only to an entity that contains the compound of the present invention, but also to any entity that is obtained directly or indirectly by combining the compound of the present invention with other ingredients. The composition of the present invention can be used, for example, as a research reagent in vitro or in vivo. The composition of the present invention can also be applied directly to the skin of a human or non-human subject for cosmetic effects.

[0142] The compositions of the present invention may be administered in any desired effective manner, for oral ingestion or parenteral, or any other appropriate mode of administration, such as intraperitoneal, subcutaneous, topical, intradermal, inhalation, pulmonary, rectal, vaginal, sublingual, intramuscular, intravenous, intraarterial, intrathecal, or intralymphatic administration. Additionally, the compositions of the present invention may be administered together with other compositions. The compositions of the present invention may be administered in any desired effective manner, for oral ingestion or parenteral, or any other appropriate mode of administration, such as intraperitoneal, subcutaneous, topical, intradermal, inhalation, pulmonary, rectal, vaginal, sublingual, intramuscular, intravenous, intraarterial, intrathecal, or intralymphatic administration. , if desired, may be encapsulated or otherwise protected from gastric or other secretions.

[0143] The compositions of the present invention comprise one or more active ingredients in admixture with one or more cosmetically or pharma- ceutically acceptable carriers, and optionally one or more other compounds, ingredients, and / or materials. Regardless of the route of administration selected, the compounds and compositions of the present invention are formulated into cosmetically or pharma- ceutically acceptable dosage forms in the conventional manner known to those skilled in the art.

[0144] Cosmetically or pharma- ceutically acceptable vehicles, diluents, and carriers are well known in the art and include materials suitable for contact with human and non-human tissues without undue toxicity, incompatibility, instability, irritation, allergic reactions, and the like. Cosmetically or pharma-ceutically acceptable vehicles, diluents, and carriers include, for example, any substantially non-toxic substances that can be conventionally used for topical, oral, peritoneal, or subcutaneous administration of cosmetics or pharmaceuticals, and that remain stable and bioavailable when the compounds and compositions of the present invention are applied, ingested, injected, or otherwise administered to a human or non-human subject. Cosmetically or pharma-ceutically acceptable carriers suitable for topical administration are known to those skilled in the art and include cosmetically or pharma-ceutically acceptable liquids, creams, oils, lotions, ointments, gels, or solids, such as conventional cosmetic night creams, foundation creams, sunscreen lotions, sunscreens, hand lotions, facial cosmetics and facial makeup primers, masks, and the like. Carriers suitable for a selected dosage form and intended route of administration are well known in the art, and acceptable carriers for a selected dosage form and method of administration can be determined using ordinary skill in the art.

[0145] The compositions of the present invention may contain other ingredients conventional in cosmetics, including fragrances, estrogens, vitamins A, C, and E, alpha-hydroxy acids or alpha-keto acids, such as pyruvic acid, lactic acid, or glycolic acid, lanolin, petrolatum, aloe vera, methyl or propyl paraben, dyes, etc. Non-limiting cosmetically or pharma- ceutically acceptable vehicles, diluents, and carriers of the present invention include sugars (e.g., lactose, sucrose, mannitol, and sorbitol), starches, cellulose preparations, calcium phosphates (e.g., dicalcium phosphate, tricalcium phosphate, and calcium hydrogen phosphate), sodium citrate, water, aqueous solutions (e.g., saline, sodium chloride injection, Ringer's solution, dextrose injection, dextrose and sodium chloride injection, lactated Ringer's solution), alcohols (e.g., ethyl alcohol, propyl alcohol, and benzaldehyde injection), and the like. alcohols), polyols (e.g., glycerol, propylene glycol, and polyethylene glycol), organic esters (e.g., ethyl oleate, and triglycerides), biodegradable polymers (e.g., polylactide-polyglycolide, poly(orthoesters), and poly(anhydrides)), elastic matrices, liposomes, microspheres, oils (e.g., corn oil, germ oil, olive oil, castor oil, sesame oil, cottonseed oil, and peanut oil), cocoa butter, waxes (e.g., suppository waxes), paraffin, silicones, talc, salicylates, and the like.

[0146] The compositions of the present invention may optionally contain additional ingredients and / or materials commonly used in cosmetic compositions. These ingredients and materials are well known in the art and include, for example, (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; (2) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, hydroxypropylmethylcellulose, sucrose, and gum arabic; (3) humectants, such as glycerol; (4) disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, sodium starch glycolate, crosslinked carboxymethylcellulose, and the like. (5) solution retarding agents, such as paraffin; (6) absorption enhancers, 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, and sodium lauryl sulfate; (10) suspending agents, such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, fine (11) buffering agents; (12) excipients, such as lactose, milk sugar, polyethylene glycol, animal and vegetable fats, oils, waxes, paraffin, cocoa butter, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicones, bentonite, silicic acid, talc, salicylates, zinc oxide, aluminum hydroxide, calcium silicate, and polyamide powders; (13) inert diluents, such as water or other solvents; (14) antibacterial agents, such as cellulose acetate, cellulose acetate phosphate ... phosphate, cellulose acetate phosphate phosphate phosphate phosphate, cellulose acetate phosphate phosphate phosphate phosphate, cellulose acetate phosphate phosphate phosphate phosphate phosphate, cellulose acetate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate phosphate (15) surfactants; (16) dispersing agents; (17) controlled release or absorption retarding agents, such as hydroxypropyl methylcellulose, other polymer matrices, biodegradable polymers, liposomes, microspheres, aluminum monostearate, gelatin, and waxes; (18) opacifying agents; (19) adjuvants; (20) wetting agents; (21) emulsifying and suspending agents; (22) solubilizing and emulsifying agents, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, proline, tert-butyl ether ... propylene glycol, 1,3-butylene glycol, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan; (23) propellants, such as chlorofluorohydrocarbons, and volatile unsubstituted hydrocarbons, such as butane and propane; (24) antioxidants; (25) agents to render the formulation osmotic pressure the same as the blood of the intended recipient, such as sugars and sodium chloride; (26) thickening agents;(27) coating materials, e.g., lecithin; and (28) sweeteners, flavorings, colorings, perfumes, and preservatives. Each such ingredient or material must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the subject. Ingredients and materials suitable for a selected dosage form and intended route of administration are well known in the art, and ingredients and materials acceptable for a selected dosage form and method of administration can be determined using ordinary skill in the art.

[0147] Compositions of the invention suitable for oral administration may be in the form of a capsule, cachet, pill, tablet, powder, granule, liquid, or suspension in an aqueous or non-aqueous liquid, an oil-in-water or water-in-oil liquid emulsion, an elixir or syrup, a pastille, a bolus, a electuary, or a paste. These formulations can be prepared by methods known in the art, for example, by conventional pan-coating, mixing, granulating, or lyophilizing processes.

[0148] Solid dosage forms for oral administration (capsules, tablets, pills, dragees, powders, granules, etc.) can be prepared, for example, by mixing the active ingredient(s) with one or more cosmetically or pharma- ceutical acceptable carriers, and optionally one or more fillers, extenders, binders, humectants, disintegrants, dissolution retarders, absorption accelerators, wetting agents, absorbents, lubricants, and / or colorants. Solid compositions of a similar type may also be used as fillers in soft and hard-filled gelatin capsules, using suitable excipients. Tablets may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using suitable binders, lubricants, inert diluents, preservatives, disintegrants, surfactants, or dispersing agents. Molded tablets may be made by molding in a suitable machine. Tablets, as well as other solid dosage forms such as capsules, pills, and granules, can optionally be scored or enterically coated and other suitable formulation methods well known in the cosmetic formulation art. They may be prepared with coatings and shells such as coatings. They may also be formulated to provide sustained or controlled release of the active ingredient therein. They may be sterilized, for example, by filtration through a bacteria-retaining filter. These compositions may also optionally contain opacifying agents and may be of a composition such that they release the active ingredient only, or preferentially, in a certain part of the gastrointestinal tract, optionally in a delayed manner. The active ingredient may also be in microencapsulated form.

[0149] Liquid dosage forms for oral administration include cosmetic or pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.Liquid dosage forms may contain suitable inert diluents that are commonly used in the art.Besides inert diluents, oral compositions may also contain adjuvants, such as wetting agents, emulsifying and suspending agents, sweeteners, flavoring agents, coloring agents, perfumes and preservatives.Suspensions may contain suspending agents.

[0150] The compositions of the present invention for rectal or vaginal administration may be provided in suppositories, which may be prepared by mixing one or more active ingredient(s) with one or more suitable non-irritating carriers that are solid at room temperature but liquid at body temperature and thus melt in the rectal or vaginal cavity and release the active compound. Compositions of the present invention suitable for vaginal administration also include suppositories, tampons, creams, gels, pastes, foams, or sprays containing cosmetic or pharma- ceutically acceptable carriers as known in the art as appropriate.

[0151] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, drops, emulsions, suspensions, aerosols, and inhalants. Any desired conventional vehicles, adjuvants, and optionally further active ingredients may be added to the formulations.

[0152] Preferred auxiliaries are from the group comprising preservatives, antioxidants, stabilizers, solubilizers, vitamins, colorants, odour improvers, film-formers, thickeners and moisturizers.

[0153] The solutions and emulsions can contain conventional vehicles such as solvents, solubilizers, and emulsifiers, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol, oils, particularly cottonseed oil, peanut oil, maize (corn) oil, olive oil, castor oil, and sesame oil, glycerol fatty acid esters, polyethylene glycols, and fatty acid esters of sorbitan, or mixtures of these substances.

[0154] Emulsions may exist in various forms: they can be, for example, water-in-oil (W / O) or oil-in-water (O / W) emulsions, or multiple emulsions, for example water-in-oil-in-water (W / O / W) emulsions or microemulsions.

[0155] The compositions according to the invention may also be in the form of emulsifier-free dispersed preparations. They may be, for example, aqueous dispersions or Pickering emulsions.

[0156] Suspensions may contain conventional vehicles, such as liquid diluents (e.g., water, ethanol, or propylene glycol), suspension media (e.g., ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, and tragacanth), or mixtures of these substances.

[0157] The pastes, ointments, gels, and creams may contain customary vehicles, such as animal and vegetable fats, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc, and zinc oxide, or mixtures of these substances.

[0158] Face and body oils may contain conventional vehicles such as synthetic oils (fatty acid esters, fatty alcohols, silicone oils, etc.), natural oils (vegetable oils and oily plant extracts, paraffin oil, lanolin oil, etc.), or mixtures of these substances.

[0159] Sprays can contain customary propellants, such as chlorofluorocarbons, propane / butane, or dimethyl ether.

[0160] The compositions of the present invention suitable for parenteral administration include one or more compounds in combination with one or more cosmetic or pharma- ceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions, or sterile powders that can be reconstituted immediately before use into sterile injectable solutions or dispersions, which may contain suitable antioxidants, buffers, solutes that give the formulation the same osmotic pressure as the blood of the intended recipient, or suspending or thickening agents. Proper fluidity can be maintained, for example, by the use of coating materials, by maintaining the required particle size in the case of dispersions, and by the use of surfactants. These compositions may also contain suitable adjuvants, such as wetting agents, emulsifying agents, and dispersing agents. It may be desirable to include an isotonic agent. In addition, sustained absorption of the injectable cosmetic dosage form may be achieved by including an agent that delays absorption.

[0161] In some cases, in order to prolong the effect, it is desirable to slow the absorption from subcutaneous or intramuscular injection This can be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility.

[0162] The rate of absorption of the active agent / drug then depends on its dissolution rate, which may further depend on the crystal size and crystalline form. Alternatively, delayed absorption of parenterally administered compositions may be achieved by dissolving or suspending the active composition in an oil vehicle. Injectable depot dosage forms may be prepared by forming microencapsule matrices of the active ingredient in biodegradable polymers. Depending on the ratio of active ingredient to polymer and the nature of the particular polymer used, the release rate of the active ingredient can be controlled. Depot injectable formulations can also be prepared by encapsulating the drug in liposomes or microemulsions that are compatible with body tissues. Injectable materials can be sterilized, for example, by filtration through a bacteria-retaining filter.

[0163] The compositions of the invention may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a freeze-dried condition requiring only the addition of the sterile liquid carrier, for example, water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets of the type described above.

[0164] In the present invention, the term "crystal form" refers to the crystal structure of a compound. A compound may exist in one or more crystal forms, which may have different structural, physical, pharmacological, or chemical properties. Differences in nucleation, growth kinetics, aggregation, and breakage can be used to obtain different crystal forms. Nucleation occurs when a phase transition energy barrier is overcome, thereby allowing particles to form from a supersaturated solution. Crystal growth is the enlargement of crystal particles caused by the deposition of chemicals on the existing surface of the crystal. The relative rates of nucleation and growth determine the size distribution of the crystals formed. Thermodynamic driving forces of nucleation and growth Both are supersaturated, which is defined as a departure from thermodynamic equilibrium. Agglomeration is the formation of larger particles by two or more particles (e.g., crystals) sticking together to form a larger crystalline structure.

[0165] The term "hydrate" as used herein means a solid or semi-solid form of a chemical that contains water in a molecular complex. The water is generally in a stoichiometric amount to the chemical.

[0166] As used herein, "cosmetic or pharmaceutically acceptable salt" refers to a derivative of the compound disclosed herein, which is modified by making an acid or base salt thereof. Cosmetic or pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines, alkali or organic salts of acid residues such as carboxylic acids, and the like.For example, such salts include ammonia, L-arginine, betaine, benethamine, benzathine, calcium hydroxide, choline, deanol, diethanolamine (2,2'-iminobis(ethanol)), diethylamine, 2-(diethylamino)-ethanol, 2-aminoethanol, ethylenediamine, N-ethyl-glucamine, hydrabamine, 1H-imidazole, lysine, magnesium hydroxide, 4-(2-hydroxyethyl)-morpholine, piperazine, potassium hydroxide, 1-(2-hydroxyethyl)-pyrrolidine, sodium hydroxide, triethanolamine (2,2',2"-nitrilotris(ethanol)), tromethamine, , zinc hydroxide, acetic acid, 2.2-dichloro-acetic acid, adipic acid, alginic acid, ascorbic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid, 2,5-dihydroxybenzoic acid, 4-acetamido-benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, decanoic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxy-ethanesulfonic acid, ethylenediaminetetraacetic acid, formic acid, fumaric acid, galactacrucic acid, gentisic acid, D-glucoheptonic acid, D-gluconic acid, D-glucuronic acid, glutamic acid, glutantic acid acid), glutaric acid, 2-oxo-glutaric acid, glycero-phosphoric acid, glycine, glycolic acid, hexanoic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, DL-lactic acid, lactobionic acid, lauric acid, lysine, maleic acid, (-)-L-malic acid, malonic acid, DL-mandelic acid, methanesulfonic acid, galactaric acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy- Included are salts from ci-2-naphthoic acid, nicotinic acid, nitric acid, octanoic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid (embonic acid), phosphoric acid, propionic acid, (-)-L-pyroglutamic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, and undecylenic acid.Additional cosmetically or pharma- ceutical acceptable salts can be formed with cations from metals such as aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, and the like.

[0167] The cosmetic or pharma- ceutically acceptable salts of the present invention can be synthesized from the compounds disclosed herein that contain a basic or acidic moiety by conventional chemical methods. In general, such salts can be prepared by reacting the compounds in their free acid or free base form with a sufficient amount of the appropriate base or acid in water or in an organic diluent such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile, or a mixture thereof.

[0168] It is contemplated that the compounds and compositions of the present invention may be included in cosmetic or pharmaceutical compositions.

[0169] The terminology used herein is for the purpose of describing particular embodiments only. As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0170] When numerical ranges are recited herein, it is expressly intended that each numerical value exists therebetween with the same precision. For example, in the range of 6 to 9, the numerical values ​​of 7 and 8 are expressly intended in addition to 6 and 9, and in the range of 6.0 to 7.0, the numerical values ​​of 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are expressly intended.

[0171] The following examples are provided to further illustrate the method of the present invention. These examples are illustrative only and are not intended to limit the scope of the invention in any way. EXAMPLES

[0172] Example 1 Materials and Methods Isolation of compounds produced by Malassezia Malassegins are isolated using procedures outlined, for example, in Wille et al. 2001. The protocol is briefly outlined below.

[0173] Culture medium Sterilize medium consisting of Tween 80 (30 mL), cycloheximide (0.5 g), chloramphenicol (0.05 g), agar (20 g), and enough water to make a 1000 mL mixture with sterile filtered 0.3% L-tryptophan at a concentration of 0.3 g% at 50° C. Pour 10 mL into 10 cm Petri dishes and adjust the pH to 5.5 using 0.1 M HCl.

[0174] Cultivation of Malassezia furfur and isolation of compounds produced by M. furfur. Malassezia furfur is swabbed onto the above medium and incubated at 30°C for 14 days. The contents of the Petri dish are pureed and extracted with ethyl acetate for 12 hours. The extract is filtered through glass wool, evaporated to dryness and dissolved in methanol. The extract is then fractionated by chromatography on Sephadex LH-20 using methanol as eluent. Further separation is performed by preparative thin layer chromatography using toluene:ethyl formate:formic acid (10:5:3). The main zone is partitioned between water and ethyl acetate. The fractions are analyzed for the activity of interest. The compound is isolated from the fraction of interest by HPLC.

[0175] Synthesis of malassedin and chemical analogues of malassedin. Malassedin is synthesized according to the protocol described in Wille et al. 2001. Chemical analogs of malassedin are synthesized according to novel synthetic protocols as well as protocols described in Winston-McPherson, et al. 2014.

[0176] Screening Protocol Both screening protocols known to those skilled in the art and novel screening methods are used to evaluate effective skin lightening compounds. For example, malassezins and their chemical analogs are evaluated by the tyrosinase bioassay as described below. Other screening protocols include both in vitro cell models and in vivo tissue models, such as the aryl hydrocarbon receptor (AhR) binding assay.

[0177] Tyrosinase bioassay Tyrosinase bioassay is carried out as described in Wille et al. 2001. Briefly, L-DOPA is mixed with tyrosinase enzyme. The absorbance is measured over 1 minute to indicate the formation of dopaquinone. For example, when using the fractions discussed above, these fractions are dissolved in DMSO and added directly to the tyrosinase reaction, with pure DMSO as the control. Tyrosinase inhibitory activity is measured as the reduction in the increase in absorbance compared to the control.

[0178] Aryl hydrocarbon receptor binding assay AhR binding assays are performed according to the protocol described, for example, in Song, et al., 2002. Briefly, human and mouse AhR are expressed in vitro, for example, using the TnT Quick-coupled Reticulocyte Lysate Systems reaction (Promega, Madison, WI). Receptor-ligand binding studies utilize sucrose density gradient velocity sedimentation as described in Karchner, et al., 1999.

[0179] EROD assay The compounds, compositions and formulations of the invention are also evaluated using the ethoxyresorufin-O-deethylase (EROD) assay known to those of skill in the art (Donato, et al., 1993; Whyte, et al., 2000; Wille et al., 2001).

[0180] Melanocyte apoptosis assay Candidate compounds are evaluated for apoptosis-inducing activity in melanocytes. Human epidermal melanocytes are cultured in Medium 254 (Thermo-Fisher Scientific, Waltham, MA) or Dermal Cell Basal Medium (ATCC, Manassas, VA) supplemented with Human Melanocyte Growth Supplement (HMGS). Additional components of human melanocyte growth medium may include, but are not limited to, insulin (5 μg / mL), ascorbic acid (50 μg / mL), L-glutamine (6 mM), epinephrine (1.0 μM), and calcium chloride (0.2 mM). Human melanocyte cultures are maintained at 4°C for 24 hours at 37°C (28°F) in a 5% CO 2 The incubation was continued at 37°C.

[0181] Candidate compounds are diluted in DMSO and mixed directly into melanocyte cultures. The same amount of pure DMSO is used as a control. Cytotoxicity assays known to those skilled in the art are performed according to the manufacturer's instructions. Cytotoxicity assays used in the present invention include, but are not limited to, CellTox™ Green Cytotoxicity Assay, Apo-ONE fluorescent caspase assay, ApoTox-Glo™ assay, and Caspase-Glo® assay (Promega, Madison, WI). Fluorescence detection is performed using standard FACS or microscopic assays known to those skilled in the art, including those described in Kramer, et al., 2005.

[0182] Additional means of assessing apoptosis are used, including FACS analysis for Annexin V, and Western blot for caspase-9 expression. Western blots are performed according to methods known to those of skill in the art.

[0183] Mouse xenograft assay Mouse xenograft models of human skin are generated according to protocols known in the art (Black, et al., 1985; Manning et al., 1973; Reed, et al., 1973; Plenat, et al., 1992; Scott et al., 1998; Otulakowski, et al., 1994). After establishing a mouse xenograft model, it is exposed to the compound of the present invention and observed for changes in pigmentation compared to the control. Changes in skin pigmentation are evaluated using various pigmentation scales known to those skilled in the art. These include, but are not limited to, the Fitzpatrick skin classification test and the Taylor hyperpigmentation scale (Taylor, et al., 2005).

[0184] Human Assays The compounds, compositions and formulations of the present invention are applied to humans, for example to human skin, and compared to a control substance. Changes in skin pigmentation are assessed using various pigmentation scales known to those skilled in the art, including, but not limited to, the Fitzpatrick skin classification test and the Taylor hyperpigmentation scale.

[0185] Example 2 Biochemical targets of malassezin and its analogues. The compounds and compositions of the present invention are expected to exhibit tyrosinase inhibition and AhR agonist activity comparable to, for example, malassezin.The compounds and compositions of the present invention are expected to exhibit, for example, more potent tyrosinase inhibition and more potent AhR agonism compared to malassezin.Similarly, some of the compounds and compositions of the present invention are expected to be less effective tyrosinase inhibitors and AhR agonists than malassezin.Such compounds, compositions and formulations can have more favorable toxicity profiles compared to more potent compounds.

[0186] Example 3 In vitro efficacy The compounds and compositions of the present invention are expected to induce melanocyte apoptosis and modulate melanocyte activity, melanin production, melanosome biogenesis and / or melanosome transport with at least as high potency as malassezins. It is also contemplated that some of the compounds and compositions of the present invention will have less potent effects on these biological processes than malassezins. Such compounds and compositions may have a more favorable toxicity profile compared to more potent species.

[0187] Example 4 In vivo efficacy The compounds and compositions of the present invention are expected to be at least as effective as malassezin in lightening skin and improving hyperpigmentation caused by hyperpigmentation disorders.Furthermore, the compounds and compositions of the present invention are expected to show favorable pharmacokinetic profiles, for example, in terms of half-life and absorption.Some compounds will show long half-lives, while others will show short half-lives.Similarly, some compounds are expected to show different absorption profiles, with some taking longer to be completely absorbed and others taking less time to be completely absorbed.

[0188] Example 5 Synthesis of malasedin and malasedin derivatives. Malasedin ("CV-8684") and its cyclized derivative, indolo[3,2-b]carbazole ("CV-8685"), were synthesized according to the scheme shown in Figure 2A.

[0189] Synthesis of tert-butyl (2-iodo-phenyl)carbamate, compound 1 To a solution of 2-iodo-aniline (25.0 g, 0.114 mol) in tetrahydrofuran (250 mL) at 0° C., LiHMDS (251.0 mL, 1 M in THF, 0.251 mol) was added slowly over 40 min while maintaining the internal temperature below 5° C. After stirring at 0° C. for 30 min, a solution of BOC anhydride (27.0 g, 0.125 mol) in THF (50 mL) was added slowly over 40 min, maintaining the internal temperature below 5° C. The reaction mixture was allowed to warm to ambient temperature and stirred for 1 h. Saturated NH 4 The reaction was quenched by the addition of Cl (250 mL). The organic layer was separated and washed with water (150 mL). The combined aqueous layers were extracted with ethyl acetate (2×150 mL) and the layers were separated. The ethyl acetate layer was combined with the first organic layer and concentrated under reduced pressure to give a brown oil. The crude compound was purified by column chromatography (0-5% ethyl acetate / hexanes). Compound 1 was obtained as a pale yellow liquid (29.0 g, 80%).

[0190] Synthesis of compound 2 Copper iodide (0.95 g, 10% mol) and PdCl 2 (PPh 3 ) 4 (1.75 g, 5% mol) was added to a solution of compound 1 (16.0 g, 0.05 mol) and propargyl methyl ether (4.25 g, 0.06 mol) in degassed triethylamine (200 mL) at ambient temperature. After stirring at ambient temperature for 2 h, the reaction was complete (monitored by TLC using 10% ethyl acetate / hexane). The reaction mixture was diluted with ethyl acetate (300 mL) and the reaction mixture was washed with water, saturated NaCl, and Na 2 SO 4 The mixture was dried over 1000 ml of ethyl acetate and concentrated under reduced pressure to give a brown oil. The crude compound was purified by column chromatography (10% ethyl acetate / hexanes). Compound 2 was obtained as a pale yellow liquid (13.0 g, 99%).

[0191] Synthesis of compound 3 In an oven-dried flask, add PtCl 2 (0.26g, 0.001mol), Na 2 CO 3(1.6 g, 0.015 mol), indole (2.32 g, 0.02 mol), and compound 2 (2.6 g, 0.01 mol) in dioxane (120 mL) were added. The flask was degassed with nitrogen, sealed, and heated to 100° C. overnight. After the reaction was complete (monitored by TLC using 10% ethyl acetate / hexane), the solvent was removed under reduced pressure. The reaction mixture was diluted with ethyl acetate (200 mL) and the reaction mixture was washed with water, saturated NaCl, and Na 2 SO 4 The solvent was filtered and concentrated in vacuo to give a brown oil.

[0192] The reaction was repeated using another batch of compound 2 (2.6 g, 0.01 mol). The crude compounds from both batches were combined and purified by column chromatography (10% ethyl acetate / hexane). Compound 3 was obtained as a light brown solid (3.8 g, 55%).

[0193] Synthesis of compound 4 To a solution of compound 3 (3.8 g, 0.0109 mol) in a mixture of methanol (150 mL) and water (50 mL) was added potassium carbonate (4.6 g, 0.0329 mol) at ambient temperature. The resulting suspension was heated to reflux overnight. After the reaction was complete (monitored by TLC using 20% ​​ethyl acetate / hexane), the reaction mixture was cooled to ambient temperature and the solvent was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (200 mL), washed with water and brine, then dried (sodium sulfate), filtered, and the solvent was concentrated under reduced pressure to give a brown solid. The crude compound was purified by column chromatography (20% ethyl acetate / hexane). Compound 4 was obtained as an orange solid (2.2 g, 81%).

[0194] Synthesis of the compound Malasedin (CV-8684) To a dry 100 mL two-neck round bottom flask under argon at 0 °C was added dimethylformamide (20 mL). POCl was added while maintaining the internal temperature below 5 °C. 3(0.75 g, 0.0048 mol) was added slowly over 10 min. After stirring at 0° C. for 30 min, a solution of compound 4 (1.0 g, 0.004 mol) in dimethylformamide (5 mL) was added slowly over 10 min, maintaining the internal temperature below 5° C. The resulting mixture was stirred at ambient temperature overnight. After the reaction was complete (monitored by TLC using 20% ​​ethyl acetate / hexanes), the reaction mixture was poured into saturated aqueous sodium bicarbonate (150 mL) and stirred for 1 h. The resulting mixture was extracted with ethyl acetate (2×100 mL). The organic layers were combined and washed with water, saturated NaCl, and sodium bicarbonate. 2 SO 4 The crude compound was purified by column chromatography (0-20% ethyl acetate / hexanes). The compound Malasedin (CV-8684) was obtained as a pale pink solid (0.82 g, 74%).

[0195] HPLC purity: 97.8% (area %). 1 H-NMR, 13 C spectrum is consistent with the structure. ESI-MS:C 18 H 15 N 2 O (M + H) + Calculated value: 275, measured value: 275.2

[0196] Synthesis of the compound indolo[3,2-b]carbazole (CV-8685) Concentrated hydrochloric acid (0.25 mL) was added to a solution of malasedin (0.75 g) in tetrahydrofuran (120 mL) at ambient temperature. The resulting mixture was heated to reflux overnight. After the reaction was complete (monitored by TLC using 40% ethyl acetate / hexane), the reaction mixture was cooled to ambient temperature and stirred for 1 h. The solid was filtered, washed with tetrahydrofuran (20 mL), and dried to give a pale yellow solid of indolo[3,2-b]carbazole (CV-8685) (0.55 g, 78%).

[0197] HPLC purity: 96.22% (area %). 1 H-NMR, 13 C spectrum is consistent with the structure. ESI-MS:C 18 H 13 N 2 (M + H) + Calculated value: 257, measured value: 257.5.

[0198] Compound I ("CV-8686") and compound IV ("CV-8687") were synthesized according to the scheme shown in Figure 2B.

[0199] Synthesis of compound 5 In an oven-dried flask, add PtCl 2 (1.0g, 0.0038mol), Na 2 CO 3 (6.1 g, 0.057 mol), 6-methylindole (10.0 g, 0.076 mol), and compound 2 (10.0 g, 0.038 mol) in dioxane (250 mL) were added. The flask was degassed with nitrogen, sealed, and heated to 100° C. overnight. After the reaction was complete (monitored by TLC using 10% ethyl acetate / hexane), the solvent was removed under reduced pressure. The reaction mixture was diluted with ethyl acetate (400 mL) and the reaction mixture was washed with water, saturated NaCl, and Na 2 SO 4 The mixture was dried over 1000 ml of ethyl acetate and concentrated under reduced pressure to give a brown oil. The crude compound was purified by column chromatography (10% ethyl acetate / hexanes). Compound 5 was obtained as a light brown solid (6.5 g, 47%).

[0200] Synthesis of compound 6 To a solution of compound 5 (6.5 g, 0.018 mol) in a mixture of methanol (150 mL) and water (50 mL) was added potassium carbonate (7.4 g, 0.054 mol) at ambient temperature. The resulting suspension was heated to reflux overnight. After the reaction was complete (monitored by TLC using 20% ​​ethyl acetate / hexane), the reaction mixture was cooled to ambient temperature and the solvent was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (200 mL), washed with water and brine, then dried (sodium sulfate), filtered, and the solvent was concentrated under reduced pressure to give a brown solid. The crude compound was purified by column chromatography (20% ethyl acetate / hexane). Compound 6 was obtained as an orange solid (3.3 g, 72%).

[0201] Synthesis of compound I (CV-8686) To a dry 100 mL two-neck round bottom flask under argon at 0 °C was added dimethylformamide (20 mL). POCl was added while maintaining the internal temperature below 5 °C. 3 (0.6 g, 0.0038 mol) was added slowly over 10 min. After stirring at 0° C. for 30 min, a solution of compound 6 (1.0 g, 0.0038 mol) in dimethylformamide (5 mL) was added slowly over 10 min, while maintaining the internal temperature below 5° C. The resulting mixture was stirred at ambient temperature overnight. After the reaction was complete (monitored by TLC using 20% ​​ethyl acetate / hexane), the reaction mixture was poured into saturated aqueous sodium bicarbonate (150 mL) and stirred for 1 h. The resulting mixture was extracted with ethyl acetate (2×100 mL). The organic layers were combined, washed with water, saturated NaCl, and diluted with Na 2 SO 4 The mixture was dried over low heat. The solvent was filtered and concentrated under reduced pressure to give a brown solid. The crude compound was purified by column chromatography (0-20% ethyl acetate / hexane). Compound I (CV-8686) was obtained as a pale pink solid (0.84 g, 75%).

[0202] HPLC purity: 97.01% (area %). 1 H-NMR, 13C spectrum is consistent with the structure. ESI-MS:C 19 H 17 N 2 O (M + H) + Calculated value: 289, measured value: 289.1

[0203] Synthesis of compound IV (CV-8687) Concentrated hydrochloric acid (0.3 mL) was added to a solution of compound I (1.0 g) in tetrahydrofuran (125 mL) at ambient temperature. The resulting mixture was heated to reflux overnight. After the reaction was completed (monitored by TLC using 40% ethyl acetate / hexane), the reaction mixture was cooled to ambient temperature and stirred for 1 hour. The solid was filtered, washed with tetrahydrofuran (20 mL), and dried to give a pale yellow solid of compound IV (CV-8687) (0.84 g, 89%).

[0204] HPLC purity: 98.4% (area %). 1 H-NMR, 13 C spectrum is consistent with the structure. ESI-MS:C 19 H 15 N 2 (M + H) + Calculated value: 271, measured value: 271.3.

[0205] Compound II ("CV-8688") was synthesized according to the scheme shown in Figure 2C.

[0206] Synthesis of compound 7 Copper iodide (0.53 g, 10% mol) and PdCl 2 (PPh 3 ) 4 (1.0 g, 5% mol) was added to a solution of compound 1 (9.0 g, 0.03 mol) and 3-methoxy-1-butyne (2.8 g, 0.035 mol) in degassed triethylamine (150 mL) at ambient temperature. After stirring at ambient temperature for 2 h, the reaction was complete (monitored by TLC using 10% ethyl acetate / hexane). The reaction mixture was diluted with ethyl acetate (300 mL) and the reaction mixture was washed with water, saturated NaCl, and Na 2 SO4 The mixture was dried over 1000 ml of ethyl acetate and concentrated under reduced pressure to give a brown oil. The crude compound was purified by column chromatography (10% ethyl acetate / hexanes). Compound 7 was obtained as a pale yellow liquid (7.0 g, 90%).

[0207] Synthesis of compound 8 In an oven-dried flask, add PtCl 2 (0.68g, 0.0025mol), Na 2 CO 3 (4.0 g, 0.038 mol), indole (6.0 g, 0.05 mol), and compound 7 (10.0 g, 0.025 mol) in dioxane (250 mL) were added. The flask was degassed with nitrogen, sealed, and heated to 100° C. overnight. After the reaction was complete (monitored by TLC using 10% ethyl acetate / hexane), the solvent was removed under reduced pressure. The reaction mixture was diluted with ethyl acetate (400 mL), and the reaction mixture was washed with water, saturated NaCl, and Na 2 SO 4 The solvent was filtered and concentrated under reduced pressure to give , was obtained as a brown oil. The crude compound was purified by column chromatography (10% ethyl acetate / hexanes). Compound 8 was obtained as a light brown solid (3.5 g, 77%).

[0208] Synthesis of compound 9 To a solution of compound 8 (3.3 g, 0.0091 mol) in a mixture of methanol (75 mL) and water (25 mL) was added potassium carbonate (3.8 g, 0.027 mol) at ambient temperature. The resulting suspension was heated to reflux overnight. After the reaction was complete (monitored by TLC using 20% ​​ethyl acetate / hexane), the reaction mixture was cooled to ambient temperature and the solvent was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (200 mL), washed with water and brine, then dried (sodium sulfate), filtered, and the solvent was concentrated under reduced pressure to give a brown solid. The crude compound was purified by column chromatography (20% ethyl acetate / hexane). Compound 9 was obtained as an orange solid (2.1 g, 88%).

[0209] Synthesis of compound II (CV-8688) To a dry 100 mL two-neck round bottom flask under argon at 0 °C was added dimethylformamide (20 mL). POCl was added while maintaining the internal temperature below 5 °C. 3 (0.76 g, 0.005 mol) was added slowly over 10 min. After stirring at 0° C. for 30 min, a solution of compound 9 (1.3 g, 0.005 mol) in dimethylformamide (5 mL) was added slowly over 10 min, while maintaining the internal temperature below 5° C. The resulting mixture was stirred at ambient temperature overnight. After the reaction was complete (monitored by TLC using 20% ​​ethyl acetate / hexane), the reaction mixture was poured into saturated aqueous sodium bicarbonate (150 mL) and stirred for 1 h. The resulting mixture was extracted with ethyl acetate (2×100 mL). The organic layers were combined, washed with water, saturated NaCl, and diluted with Na 2 SO 4 The mixture was dried over 1000 ml of ...

[0210] HPLC purity: 98.94% (area %). 1 H-NMR, 13 C spectrum is consistent with the structure. ESI-MS:C 19 H 17 N 2 O (M + H) + Calculated value: 289, actual value: 289.0.

[0211] Example 6 Cell morphology Typical cell morphology after various treatments is shown in Figures 5A-5K, 6A-6K, 7A-7K, 8A-8K, 9A-9K, 10A-10K, 11A-11K, and 12A-12K. Both cell lines were significantly affected in morphology by 100 μM CV-8684 and CV-8688, as well as staurosporine for 6 hours. CV-8685 was found to only affect WM115 at 100 μM.

[0212] Example 7 Apoptosis-inducing activity of malassedin and malassedin derivatives-A preliminary annexin V assay Materials and Reagents Annexin V-FITC assay kit was purchased from Beyotime Biotechnology, RPMI1640 medium and Dulbecco's modified Eagle's medium ("DMEM") were purchased from Gibco, fetal bovine serum ("FBS") was purchased from Invitrogen, stabilized antibacterial antifungal solution (100x) was purchased from Sigma, 0.25% trypsin-EDTA (1x), phenol red were purchased from Invitrogen.

[0213] cell culture MeWo (ATCC® HTB-65™) and WM115 (ATCC® CRL-1675) cells were purchased from ATCC (Manassas, VA), and MeWo was maintained in DMEM supplemented with 10% FBS, and WM115 was maintained in RPMI 1640 supplemented with 10% FBS (10% FBS, 1% stabilized antibacterial-antimycotic solution).

[0214] Test Overview During intermediate stages of apoptosis, phosphatidylserine ("PS") is transferred from the inner to the outer leaflet of the plasma membrane, exposing PS to the extracellular environment where it can be detected. Highly fluorescent annexin V conjugates provide a rapid and reliable detection method to test for PS externalization.

[0215] During the first series of tests, both MeWo and WM115 cells were treated with 10 doses of test compounds prepared by 3-fold dilution from 100 μM. Staurosporine was used as a positive control. After 6 hours of treatment, cell apoptosis was evaluated using the Annexin V assay. The test compounds evaluated were CV-8684, CV-8685, CV-8686, CV-8687 and CV-8688.

[0216] Assay procedure For cell seeding, cells were harvested and cell number was determined using a Countess® cell counter. Cells were then diluted with culture medium to the desired density. 40 μL of cell suspension was added per well to the required number of wells of a 384-well plate (Corning 3712-clear bottom plate). The final cell density was 6,000 cells / well. After plating, the plates were incubated at 37°C and 5% CO 2 The mixture was incubated overnight at 4°C.

[0217] For preparation of compound source plate, each test compound was dissolved in DMSO to obtain 10 mM stock. 3-fold serial dilutions were performed using EVO200™ liquid handler (TECAN) to generate 10 concentrations of test compound. 0.1% DMSO was used as vehicle (negative) control. Compound source plate was then rotated at 1,000 RPM for 1 minute at room temperature and shaken for 2 minutes using a plate shaker.

[0218] For compound treatment, 40 nL of compound was transferred from the compound source plate to the 384-well culture plate using a liquid handler Echo550 (LabCyte Inc.). After 6 h of incubation, the plate was removed from the incubator for detection.

[0219] For preliminary Annexin V assay, the plates were removed from the incubator and equilibrated at room temperature for 15 minutes. The medium was then removed. 20 μL of premixed dye working solution of Annexin V-FITC and Hoechst33342 was added to each well. The cells were then incubated at room temperature for 20 minutes. The plates were sealed and centrifuged at 1,000 RPM for 1 minute to remove air bubbles. The plates were then read using an Acumen eX3 plate reader. Relative activity was calculated according to the following formula: Activity (%) = 100% x (Counts) アネキシンV / Count 全細胞 ).EC 50 was calculated using GraphPad Prism (v.5.01).

[0220] result In the above preliminary screening, CV-8688 significantly increased Annexin V staining in MeWo cells and inhibited EC 50 The positive control, staurosporine, significantly increased Annexin V staining in both cell lines (Figures 3A-3M).

[0221] Example 8 Apoptosis-inducing activity of malassedin and malassedin derivatives-Further evaluation using the Annexin V assay Test Overview To further investigate the effect of test compounds on apoptosis, multiple readouts encompassing various stages of apoptosis were performed in both MeWo and WM115 cells. Both cell types were treated with three doses of test compounds (100 μM, 10 μM and 1 μM). Staurosporine was used as a positive control. After the desired treatment period (6, 24, 48 or 72 hours), apoptosis was assessed by measuring the percentage of cells showing Annexin V binding after exposure to test compounds. The test compounds evaluated were CV-8684, CV-8685 and CV-8688.

[0222] Assay procedure Cell seeding was performed as discussed above with the following exceptions: the final cell density was 4,000 cells / well for 6 and 24 hour detection, whereas 2,000 cells / well was used for 48 and 72 hour detection. 384-well clear bottom plates (Corning 3712) and white bottom plates (Corning 3570) were prepared for each time point. Plates were incubated as discussed above.

[0223] For compound source plate preparation, each test compound was dissolved in DMSO to obtain a 10 mM stock. Two additional concentrations were generated by 10-fold dilution to 1 mM and 0.1 mM. Staurosporine was used as a positive control and 1% DMSO was used as a solvent (negative) control. The compound source plate was rotated at 1,000 RPM for 1 minute at room temperature and shaken for 2 minutes using a plate shaker.

[0224] An Echo550 liquid handler was used to transfer 400 nL of test compounds from the compound source plate to the 384-well culture plate. After 6, 24, 48 and 72 hours, the plates were removed from the incubator for detection.

[0225] For the Annexin V assay, plates were removed from the incubator and equilibrated at room temperature for 15 min. The medium was removed and cells were washed twice with PBS. 20 μL of premixed Annexin V-FITC dye working solution was added to each well. Cells were incubated at room temperature for 20 min. Plates were read using an Acumen eX3 and the number of FITC positive cells was counted. Relative activity was calculated according to the following formula: Relative activity (%) = 100% x (counts) 試料 / Count ビヒクル ).

[0226] result CV-8684 induced apoptosis in both MeWo and WM115 cells at the highest tested concentration after 6 hours of treatment. We found that CV-8685 required 48 hours to induce apoptosis in both cell types, whereas CV-8685 showed an inductive effect on WM115 cells after 24 hours of treatment. Finally, CV-8688 showed a dose-dependent inductive effect on both cell types within 6 hours of treatment (Figures 4A-4L).

[0227] Example 9 Cell viability following exposure to Malassedin and Malassedin derivatives - CellTiter-Glo® Assay Assay procedure The CellTiter-Glo® 2.0 assay was purchased from Promega. Cell seeding, preparation of compound source plates, and exposure of cells to test compounds were performed as described in Example 8.

[0228] For the CellTiter-Glo® assay, plates were removed from the incubator and equilibrated at room temperature for 15 minutes. CellTiter-Glo® reagent was thawed and equilibrated to room temperature prior to the experiment. Then, 40 μL of CellTiter-Glo® reagent was added to each well (at a 1:1 ratio to culture medium) for detection. Plates were then incubated at room temperature for 30 minutes and read using an EnSpire (PerkinElmer) plate reader. Residual activity was calculated by the following formula: Residual activity (%) = 100% x (brightness) 試料 -brightness バックグラウンド ) / (brightness 溶媒 -brightness バックグラウンド ).

[0229] result CV-8684 showed a dose-dependent inhibition of cell viability in both tested cell lines, but the inhibitory effect was found to be more potent in MeWo cells. CV-8685 showed a dose-dependent inhibitory effect on WM115 cell viability only after 24 hours of treatment. CV-8688 dose-dependently inhibited the viability of both cell types. The positive control, staurosporine, exerted 100% inhibition of cell viability in both cell lines after 24 hours of treatment. (Figures 13A-13K).

[0230] Example 10 Cytotoxicity of malassezin and malassezin derivatives - lactate dehydrogenase release assay Test Overview The LDH assay quantitatively measures lactate dehydrogenase ("LDH") released into the medium from damaged cells as a biomarker of cytotoxicity and cytolysis.

[0231] Assay procedure CytoTox-ONE™ Homogenous Membrane Integrity Assay was purchased from Promega. Cell seeding, preparation of compound source plates, and exposure of cells to test compounds were performed as described in Example 8.

[0232] For the LDH release assay, the plate was removed from the incubator and equilibrated at room temperature for 15 minutes. The plate was then centrifuged at 1,000 RPM for 1 minute. 20 μL of cell culture was transferred into a new 384-well black bottom plate. 20 μL of CytoTOX-ONE™ was then added into each well and incubated at room temperature for 10 minutes. After that, 10 μL of stop solution was added into each well, and the plate was shaken at 500 rpm for 1 minute. The plate was read on an EnSpire using an excitation wavelength of 560 nm and an emission wavelength of 590 nm. Relative activity was calculated according to the following formula: Relative Activity (%) = 100% x (Brightness) 試料 -brightness バックグラウンド ) / (brightness 溶媒 -brightness バックグラウンド ).

[0233] result CV-8684 did not induce significant release in either cell line after 72 h of incubation. CV-8685 showed a dose-dependent induction effect on LDH release from WM115 cells, but not MeWo cells, after 24 h of treatment. CV-8688 induced LDH release at the highest tested concentration (Figures 14A-14L).

[0234] Example 11 Aryl hydrocarbon receptor activation ability of malassezin and malassezin derivatives Assay procedure HepG2-AhR-Luc cells were purchased from Pharmaron, the One-Glo luciferase assay system was purchased from Promega, DMEM was purchased from Hyclone, and penicillin / streptomycin was purchased from Solabio.

[0235] Culture medium for stably transfected HepG2 cells was prepared by supplementing DMEM with high glucose and L-glutamine, and 10% FBS.

[0236] HepG2-AhR-Luc cells were cultured in T-75 flasks at 37°C and 5% CO 2 and 95% relative humidity. Cells were allowed to reach 80-90% confluence before detachment and splitting.

[0237] The cultured cells were rinsed with 5 mL of PBS. The PBS was removed by aspiration, and 1.5 mL of trypsin was added to the flask and the cells were incubated at 37° C. for approximately 5 minutes or until the cells detached and lifted. Trypsin was inactivated by adding excess serum-containing medium.

[0238] The cell suspension was transferred to a conical tube and centrifuged at 120 g for 10 min to pellet the cells. The cells were resuspended in seeding medium at a suitable density. 40 μL of cells were transferred to a 384-well culture plate (5 × 10 3 cells / well). The plates were placed in a 37° C. incubator for 24 hours.

[0239] Stock solutions of test compounds and omeprazole positive control were then prepared. 40 nL of compound solution was transferred into the assay plate using the Echo550. The plate was then placed back into the incubator for compound treatment.

[0240] Later, 24 hours after treatment, the plates were removed from the incubator and allowed to cool at ambient temperature. 30 μL of One-Glo reagent, equal to that of the culture medium, was added to each well. Cells were allowed to lyse for at least 3 minutes and then measured in a luminometer.

[0241] Dose-response was graphed using nonlinear regression analysis in XLfit, and EC 50 values ​​were also calculated.

[0242] result The results of the AhR-luciferase assay are shown in Figures 15A-15F.

[0243] Example 12 MelanoDerm™ Assay Test Overview The purpose of this study is to evaluate the potential skin irritancy of test articles on the MelanoDerm™ Skin Model after repeated exposure for dose selection in subsequent studies. Toxicity is determined by measuring the relative conversion of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) in tissues treated with the test article compared to tissues treated with a negative / solvent control.

[0244] The MelanoDerm™ Skin Model provided by MatTek Corporation (Ashland, MA) is used in this study. The MelanoDerm™ tissue consists of normal human-derived epidermal keratinocytes (NHEK) and melanocytes (NHM) cultured to form a highly differentiated, multi-layered model of human epidermis. The NHMs in the co-culture undergo spontaneous melanogenesis, resulting in various levels of pigmentation in the tissue. The cultures are grown at an air-liquid interface on cell culture inserts, allowing for topical administration of skin modulators. The MelanoDerm™ model exhibits morphological and ultrastructural characteristics similar to those in vivo. NHMs, localized in the basal cell layer of the MelanoDerm™ tissue, are dendritic and spontaneously produce melanin granules, which The melanin gradually occupies the layers of tissue, and thus the test system can be used to screen for materials that can inhibit or stimulate melanin production in comparison to a negative control.

[0245] The experimental design of this study consists of a determination of the pH of the undiluted test article (and / or dosing solution, if necessary), when possible, and a definitive assay to determine relative tissue viability after repeated exposure. The MelanoDerm™ Skin Model is exposed to the test article for a total of 7 days. The test article is applied topically to the MelanoDerm™ Skin Model every 48 hours (within a time period of 48±2 hours from the previous treatment). Test article toxicity is determined by NAD(P)H-dependent microsomal enzyme reduction of MTT (and, to a lesser extent, succinate dehydrogenase reduction of MTT) in control and test article-treated tissues. (Berridge et al., 1996). Data are presented in the form of relative survival (percentage of MTT conversion compared to negative control).

[0246] material MelanoDerm™ Maintenance Medium (EPI-100-LLMM) and MelanoDerm™ Skin Model (MEL-300-A) were supplied by Mat Tek Corporation. 1% Kojic Acid (prepared in sterile deionized water) and MTT (3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) were supplied by Sigma. Dulbecco's Modified Eagle Medium (DMEM) containing 2 mM L-glutamine (MTT supplemented medium) was supplied by Quality Biological. Isopropanol was supplied by Aldrich. Ca ++ and Mg ++ Sterile CaCl2-free Dulbecco's Phosphate Buffered Saline (CMF-DPBS) was supplied by Invitrogen (or equivalent). Sterile deionized water was supplied by Quality Biological (or equivalent). DMSO was supplied by CiVenti Chem.

[0247] Assay procedure Test articles are generally tested neat or as directed by the sponsor (see Protocol Appendix 1). Ten microliters (10 μL) or 25 μL of each test article is applied directly onto the tissue to cover the top surface. Depending on the nature of the test article (liquid, gel, cream, foam, etc.), it may be necessary to use an administration device, mesh or other aid capable of spreading the test article over the surface of the tissue.

[0248] During the dosing period, dilute each test article at least 200-fold using the appropriate amount of EPI-100-LLMM (or alternate solvent as determined during solubility testing). Fresh dilutions in EPI-100-LLMM will be prepared for each dosing. The final dilution to be performed for preparation of the dosing solution will be determined from the solubility evaluation above and recorded in the study manual.

[0249] DMSO diluted as 0.5% (v / v) in EPI-100-LLMM is used as a vehicle control and is dosed onto tissues (10 μL and 25 μL volumes) according to the same procedure used for the test articles and assay controls.

[0250] Test articles are applied topically to the MelanoDerm™ tissues every 48 hours (within 48±2 hours of the previous treatment) during the 7-day study. 10 and 25 microliters of each test article are applied to each tissue, respectively. 25 microliters of positive and negative controls are applied to each tissue, respectively.

[0251] When possible, determine the pH of the undiluted liquid test substance (and / or dosing solution, if appropriate). The pH is determined using pH paper (e.g., with a pH range of 0-14 for estimation and / or a pH range of 5-10 for determining more precise values). Typical pH increments for narrower range pH paper are approximately 0.3-0.5 pH units. The maximum increment for pH paper is 1.0 pH unit.

[0252] The definitive assay includes a negative control and a positive control. MelanoDerm™ tissue designated as the assay negative control is treated with 25 μL of sterile deionized water. Tissue designated as the assay positive control is dosed with 25 microliters of 1% Kojic Acid (prepared in sterile deionized water and filtered at the time of preparation). The 1% Kojic Acid is stored in a tube covered in aluminum foil within 2 hours of preparation until use. Exposure times for the negative and positive controls are the same as those used for the test article.

[0253] It is necessary to evaluate the ability of each test article to directly reduce MTT. A 1.0 mg / mL MTT solution is prepared in MTT-supplemented medium as described below. Approximately 25 μL of test article is added to 1 mL of MTT solution and the mixture is incubated for 1-3 hours at 37°C ± 1°C in the dark. A negative control of 25 μL of sterile deionized water is tested at the same time. If the color of the MTT solution changes to blue / purple, the test article is presumed to have reduced MTT. Water-insoluble test materials may only show direct reduction (darkening) at the test article-medium interface.

[0254] An MTT direct reduction assay may have been performed previously for the test article(s) in an unrelated study. In such cases, the results of that MTT direct reduction assay may be used in the study presented here, and the original study is referenced.

[0255] Tissue exposure: At least 16 hours after initiation of culture, photographs of the two MelanoDerm™ tissues (considered untreated at day 0) are taken using a digital camera to aid in visual assessment of the degree of tissue pigmentation at time zero of the assay. The exact procedure used to collect tissue images is specified in the study manual and report. MelanoDerm™ tissues are rinsed with CMF-DPBS and blotted dry on sterile absorbent paper to remove excess liquid. After rinsing, the MelanoDerm™ tissues are transferred to the appropriate MTT-containing wells and processed in the MTT assay as described in the MTT assay section.

[0256] At least 16 hours after the start of the culture, the tissues are transferred onto a new 6-well plate containing 0.9 mL of fresh pre-warmed EPI-100-LLMM. The test is carried out over a period of 7 days. The two tissues are treated topically with 10 and 25 microliters of each test article, respectively, on day 1 and every 48 hours (within a time period of 48±2 hours from the previous treatment). The medium is refreshed every day (within a time period of 24±2 hours from the previous refeeding). That is, the tissues are transferred onto a new 6-well plate containing 0.9 mL of fresh pre-warmed EPI-100-LLMM.

[0257] The two tissues are treated topically with 25 μL of positive and negative controls, respectively, on day 1 and every 48 hours (within a time frame of 48±2 hours from the previous treatment). The medium is refreshed every day (within a time frame of 24±2 hours from the previous refeeding). The tissues are transferred onto new 6-well plates containing 0.9 mL of fresh pre-warmed EPI-100-LLMM. The tissues are incubated at 37±1° C. (standard culture conditions) in a humidified atmosphere of 5±1% CO2 in air for the appropriate exposure time.

[0258] During the dosing period, the MelanoDerm™ tissues are first gently rinsed approximately three times with approximately 500 μL of CMF-DPBS to remove any remaining test article. The tissues are then transferred to new 6-well plates containing 0.9 mL of fresh pre-warmed EPI-100-LLMM. Transfer onto plates and dose tissues with appropriate test article, negative, or positive control. Incubate tissues at 37±1°C (standard culture conditions) in a humidified atmosphere of 5±1% CO2 in air for the appropriate exposure time. Record exact rinsing procedures in the study manual.

[0259] At the end of the 7-day study, photographs of the negative or positive control and each test article dosed MelanoDerm™ tissues are taken using a digital camera to aid in the visual assessment of the degree of tissue pigmentation at the end of the assay (day 7). The exact procedures used to collect tissue images are specified in the study manual and report. Tissue viability is then determined by MTT reduction as shown below.

[0260] MTT Assay: Thaw a 10x stock of MTT prepared in PBS (filtered at time of batch preparation) within 2 h prior to use and dilute with warm MTT supplemented medium to generate a 1.0 mg / mL solution. Add 300 µL of the MTT solution to each designated well of a pre-labeled 24-well plate.

[0261] After the exposure time, each MelanoDerm™ tissue designated for the MTT assay is rinsed with CMF-DPBS and blotted dry on sterile absorbent paper to remove excess liquid. After rinsing, the MelanoDerm™ tissue is transferred to the appropriate MTT-containing well. The 24-well plate is incubated at standard conditions for 3±0.1 hours.

[0262] After 3±0.1 hours, the MelanoDerm™ tissues are blotted dry on sterile absorbent paper, excess liquid is removed, and transferred to a pre-labeled 24-well plate containing 2.0 mL of isopropanol in each designated well. The plate is covered with parafilm and stored in a refrigerator (2-8°C) until the last exposure is collected. If necessary, the plate may be stored in the refrigerator overnight (or up to 24 hours after the last exposure is collected) before extracting the MTT. The plate is then shaken at room temperature for at least 2 hours. At the end of the extraction period, the liquid in the cell culture insert is decanted into the well from which the cell culture insert was removed. The extraction solution is mixed and 200 μL is transferred to the appropriate wells of the 96-well plate. 200 μL of isopropanol is added to the wells designated as blanks. The optical density at 550 nm (OD550) of each well is measured on a Molecular Devices Vmax plate reader with AUTOMIX function.

[0263] If a test article is shown to reduce MTT, then only test article that remains bound to tissue after rinsing will result in a false MTT reduction signal, which poses a problem. To demonstrate that any test article that may remain is not acting to directly reduce MTT, a functional check is performed in the definitive assay to show that the test material is not bound to tissue and does not produce a false MTT reduction signal.

[0264] To determine whether residual test substances act to directly reduce MTT, control tissues killed by freezing are used. Frozen-killed tissues are prepared at IIVS by placing untreated MelanoDerm™ / EpiDerm™ (Melanoderm™ without melanocytes) tissues in a -20°C freezer for at least overnight, thawing to room temperature, and then refreezing. After killing, tissues can be stored indefinitely in the freezer. Frozen-killed tissues can be obtained pre-prepared from MatTek Corporation and stored in a -20°C freezer until use. To test for residual test substance reduction, dead tissues are treated with test substances in the usual manner. All assay procedures are performed in the same way as for live tissues. At least one dead control treated with sterile deionized water (negative dead control) is tested in parallel, since a small amount of MTT reduction from residual NADH and related enzymes in dead tissues is expected.

[0265] If little or no MTT reduction is observed in the dead control treated with the test article, then any MTT reduction observed in the live tissue treated with the test article can be attributed to viable cells. If there is readily detectable MTT reduction in the treated dead control (relative to the amount of live tissue treated), then additional steps must be taken to account for the chemical reduction or the test article can be considered untestable with this system. OD550 values ​​from the dead control are analyzed as described below.

[0266] Raw absorbance data is captured, saved as print files, and imported into an Excel spreadsheet. The mean OD550 value of the blank wells is calculated. The corrected mean OD550 value of the negative control(s) is determined by subtracting the mean OD550 value of the blank wells from these mean OD550 values. The corrected OD550 values ​​of the individual test article exposed and positive control exposed are determined by subtracting the mean OD550 value of the blank wells from each. All calculations are performed using an Excel spreadsheet. The algorithms discussed are performed to calculate the final endpoint analysis at the treatment group level, but the same calculations can be applied to individual replicates. Corrected test article exposure OD550 = test article exposure OD550 - blank average OD550

[0267] If a killed control (KC) is used, the following additional calculation is performed to correct for the amount of MTT directly reduced by test article residues: The raw OD550 value of the killed control treated with the negative control is subtracted from the raw OD550 value of the killed controls treated with each test article to obtain the net OD550 value of the killed controls treated with the test articles. Net OD550 of each test object KC = Raw OD550 of test object KC - Raw OD550 of negative control KC

[0268] The net OD550 value represents the amount of MTT reduced due to direct reduction by the test article residue at a particular exposure time. In general, if the net OD550 value is greater than 0.150, this net amount of MTT reduction is subtracted from the corrected OD550 value of the treated viable tissue to obtain the final corrected OD550 value. These final corrected OD550 values ​​are then used to calculate the survival rate (%) in proportion to the control. Final corrected OD550 = corrected specimen OD550 (viable) - pure OD550 specimen (KC)

[0269] Finally, the following % of control calculations are performed: Viability (%) = [(final corrected OD550 of test article or positive control) / (corrected mean OD550 of negative control)] x 100

[0270] result The results of the MelanoDerm™ assay are shown in Figures 16A-16K. Tissues treated with malassezin, Compound I, and Compound II demonstrated reduced pigmentation on day 7 of the study. Figures 17A-17K show 15x magnification images of MelanoDerm™ samples exposed to exemplary treatments.

[0271] Example 13 Zebrafish assay Assay procedure Compounds: Compounds are provided by the Study Sponsor as master stock (MS) solutions at maximal soluble concentration in water / PBS or DMSO.

[0272] Standard procedure for harvesting embryos: Phylonix AB strain zebrafish are generated by natural mating or using the Mass Embryo Production System (MEPS, Aquatic Habitats). Approximately 50 zebrafish are generated per female zebrafish. Zebrafish are kept in a fish tank at 20°C for 24 hours. Maintain at 8° C. Clean the zebrafish (remove dead ones) and sort them by developmental stage. Zebrafish do not require feeding for 6 days post fertilization (dpf) as they receive nutrition from the attached yolk sac.

[0273] Solubility of compounds: Dilute the master stock (MS) (use the highest concentration) in pure DMSO to make sub-stock solutions (SS), i.e., 10, 50, 100, 200, 300 mM, etc. Use fish water supplied by Phylonix [200 mg of Instant Ocean Sea Salt (Aquarium Salt) per liter of deionized water]. Systems); maintain pH at 6.6-7.0 with 2.5 mg / L Neutral Regulator (Seachem Laboratories Inc.); conductivity 850-950 μS] is dispensed into test containers at 4 ml per container.

[0274] Add 4 μl of each SS directly to the fish water to generate the test compound solution (TS). Example: Adding 4 μl of 10 mM SS to fish water will generate 10 μM TS; final DMSO concentration is 0.1%. Alternatively, 10 μl of SS can be added to 10 ml / container of fish water to obtain the same final TS and DMSO concentrations. For assays that tolerate up to 1% DMSO, 40 μl of SS can be used to generate 100 μM TS. If 10 ml of fish water is used, the volume of SS must be increased proportionately to obtain the same final TS and DMSO concentrations. The solutions are incubated at 28° C. for the period specified for each assay and visually inspected daily for the presence of precipitate.

[0275] Maximum Tolerable Concentration (MTC): MTC(LC 10 ) is used as a standard measure of compound lethality and is determined using 10 compound concentrations. The 10 concentrations are selected by the sponsor after determining the maximum soluble concentration of the compound.

[0276] Thirty chorionated Phylonix wild-type AB zebrafish, approximately 2 dpf, are distributed into wells of a 6-well microplate containing 4 ml / well of fish water and DMSO at concentrations ranging from 0.1 to 1% depending on the solubility of the compound.

[0277] Ten concentrations are initially tested: 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 50, 100, and 500 μM (or as far as compound solubility will allow). If necessary, further higher (up to 2000 μM) or lower (down to 0.001 μM) concentrations are tested.

[0278] Zebrafish are incubated with each concentration of test compound at 28 °C for 3 days in the dark. Non-treated and 0.1-1% DMSO-treated zebrafish are used as assay and vehicle controls. To calculate the mortality rate (%), count and remove the number of dead zebrafish every day after treatment. At 5 dpf, calculate the mortality rate (%) by counting the dead animals (=total number of dead zebrafish / 30). Note that if dead zebrafish are collapsed, the number of dead zebrafish is inferred by counting the number of live zebrafish.

[0279] To estimate the MTC, a mortality curve is generated by plotting the % mortality against the concentration using EXCEL software. Experiments are performed in triplicate to obtain the mean and SD of the MTC.

[0280] Visually evaluate the effect of compounds on zebrafish skin pigmentation: Zebrafish skin pigment cells, including xanthophores, iridophores, and melanophores (melanocytes), are derived from neural crest cells. In zebrafish, differentiated skin pigment progenitor cells express pigment at approximately 24 hpf. The focus of this study is on the expression of the black pigment, melanin, on the surface of the skin. Melanocytes are melanocytes that first appear as small black spots in the dorsal head region. As the zebrafish develops, the spots increase in number and merge to form a band that extends to the tail region. In contrast, mutant albino zebrafish show sparse skin pigmentation. Compounds are administered at 2 dpf to assess whether they halt the continuous process of embryonic pigmentation that is complete by 5 dpf. Three concentrations are tested for each compound: MTC, 50% MTC, and 25% MTC.

[0281] Thirty self-hatched Phylonix wild type AB zebrafish at 2 dpf are treated with each compound concentration for 3 days. Untreated and 0.1% DMSO treated zebrafish are used as controls. Positive control: phenylthiourea (PTU, 0.03%).

[0282] Zebrafish are visually inspected daily using a surgical light microscope. Compound and PTU treated zebrafish are compared to untreated and vehicle treated control zebrafish. The number of zebrafish showing reduced pigmentation is counted daily and expressed as a percentage (%) of animals examined. Representative images are shown. To identify optimal compound concentrations and treatment times for reducing pigmentation, kinetic curves are generated by plotting the percentage (%) of zebrafish showing reduced skin deposition versus time (dpf). Fisher's exact test is used to determine if the effect of the compound is significant (P<0.05).

[0283] Additional visual evaluation of the effect of the compounds on zebrafish skin pigmentation is performed after treatment with 0.1, 1, and 3 μM. Thirty self-hatched Phylonix wild-type AB zebrafish at 2 dpf are treated with each compound concentration for 3 days. Untreated and 0.1% DMSO-treated zebrafish are used as controls. Positive control: phenylthiourea (PTU, 0.003%). Zebrafish are visually inspected daily using a surgical light microscope. Compound and PTU-treated zebrafish are compared to untreated and vehicle-treated control zebrafish.

[0284] At 5 dpf, the number of zebrafish showing reduced pigmentation is counted and expressed as a percentage of tested animals. Representative images are shown. To identify optimal compound concentrations and treatment times for reducing pigmentation, kinetic curves are generated by plotting the percentage of zebrafish showing reduced skin deposition versus concentration. Fisher's exact test is used to determine if the effect of the compound is significant (P<0.05).

[0285] Quantify the effect of compounds on zebrafish skin pigmentation: Based on the results from the visual assessment, quantify the effect of compounds on zebrafish skin pigmentation using optimal conditions (concentration, compound treatment time).

[0286] Twenty Phylonix wild type AB zebrafish at optimal stage as determined by results from visual assessment are treated with optimal compound concentrations. Untreated and 0.1% DMSO treated zebrafish are used as controls. Positive control: phenylthiourea (PTU, 0.03%).

[0287] A SPOT camera at 2x magnification is used to capture images of the whole zebrafish in a dorsal view. The dorsal head and trunk regions are defined as regions of interest (ROI) using the selection function in Adobe Photoshop. Black skin pigmentation within the ROI is highlighted using the highlight function in Photoshop. Total pigment signal (PS) (in pixels) is determined using the histogram function in Photoshop.

[0288] If the compound affects the growth of zebrafish, the body length (L) and trunk width ( W) is smaller, thereby affecting the ROI area and the final PS. Therefore, the final signal (FS) measurements are normalized using FS=PS / L×W.

[0289] Untreated and vehicle-treated zebrafish are expected to show similar FS, demonstrating that the vehicle has no effect. Zebrafish treated with PTU are expected to show low FS, validating the assay. Compound-treated zebrafish are compared to vehicle-treated control zebrafish.

[0290] To determine if the effect of a compound is significant (P<0.05), the mean FS of compound-treated zebrafish is compared to the mean FS of vehicle-treated zebrafish using Student's t-test.

[0291] Additional quantification of compound effects on zebrafish skin pigmentation is performed following treatment with compound concentrations of 0.5 and 1.5 μM.

[0292] Twenty Phylonix wild type AB zebrafish at 2 dpf are treated with compound concentrations of 0.5 and 1.5 μM. Untreated and 0.1% DMSO treated zebrafish are used as controls. Positive control: phenylthiourea (PTU, 0.003%).

[0293] A SPOT camera at 2x magnification is used to capture images of the whole zebrafish in a dorsal view. The dorsal head and trunk regions are defined as regions of interest (ROI) using the selection function in Adobe Photoshop. Black skin pigmentation within the ROI is highlighted using the highlight function in Photoshop. Total pigment signal (PS) (in pixels) is determined using the histogram function in Photoshop.

[0294] If a compound affects the growth of zebrafish, the body length (L) will be shorter and the trunk width (W) will be smaller, thereby affecting the ROI area and the final PS, so the final signal (FS) measurements are normalized using FS=PS / L×W.

[0295] Untreated and vehicle-treated zebrafish are expected to show similar FS, confirming that the vehicle has no effect. Zebrafish treated with PTU are expected to show low FS, validating the assay. Compound-treated zebrafish are compared to vehicle-treated control zebrafish.

[0296] To determine if the effect of a compound is significant (P<0.05), the mean FS of compound-treated zebrafish is compared to the mean FS of vehicle-treated zebrafish using Student's t-test.

[0297] result The results of the visual assessment of zebrafish exposed to Compound II are shown in Figures 18A-18F and 19A-19F. A table summarizing the results from the visual assessment portion of the study is shown in Figure 20.

[0298] Quantitative assessment of regions of interest and results for zebrafish exposed to Compound II are shown in Figures 21A-21E and Figures 22A-22B.

[0299] Example 14 Stability of malassedin and malassedin derivatives in DMSO and cell culture medium. Test compounds were prepared at 100 μM in DMSO and culture medium. The solutions were incubated at room temperature for 2 hours and analyzed using LC-MS. Peak areas were used to determine the fraction of the compounds remaining in the solvent. Compounds that exhibit this effect were evaluated.

[0300] result The LC-MS results are shown in Figures 23A-23J and indicate that the compounds are stable in the culture medium after 2 hours of incubation.

[0301] literature Berridge, MV, Tan, AS, McCoy, KD, Wang, R. The Biochemical and Cellular Basis of Cell Proliferation Assays That Use Tetrazolium Salts.Biochemica 4:14-19(1996). Black, et al. Athymic Nude Mice and Human Skin Grafting. In: Maibach, et al. (eds.). Models in Dermatology Vol. 1. Karger, Basel, 1985, 228-39. Costin, G.-E., Raabe, R.Optimized in vitro pigmentation screening assay using a reconstructed three dimensional human skin model.Rom.J.Biochem.50(1),15-27(2013). Donato,et al.A Microassay for Measuring Cytochrome P450IA1 and P450IIB1 Activities in Intact Human and Rat Hepatocytes Cultured on 96-Well Plates.Anal Biochem.1993;213(1):29-33. Elmore. Apoptosis:A Review of Programmed Cell Death.Toxicologic Pathology 2007;35:495-516. Fitzpatrick,et al.The Validity and Practicality of Sun-Reactive Skin Types I Through VI.Arch Dermatol.1988;124(6):869-871. Gaitanis,et al.Skin Diseases Associated With Malassezia Yeasts:Facts and Controversies.Clinics in Dermatology 2013;31:455-463. Gueho,et al.The Genus Malassezia With Description of Four New Species.Antonie Van Leeuwenhoek 1996;69:337-55. Karchner,et al.Identification and Functional Characterization of Two Highly Divergent Aryl Hydrocarbon Receptors(AHR1 and AHR2)in the Teleost Fundulus heteroclitus.The Journal of Biological Chemistry 1999;274(47):33814-24. Kramer,et al.Malassezin,A Novel Analyst of the Aryl Hydrocarbon Receptor From The Yeast Malassezia furfur,Induces Apoptosis in Primary Human Melanocytes.ChemBioChem 2005;6:860-5. Lee,et al.Comparison of Gene Expression Profiles Between Keratinocytes,Melanocytes and Fibroblasts.Ann Dermatol.2013;25(1):35-45. Manning,et al.Maintenance of Skin Xenografts of Widely Divergent Phylogenetic Origin on Congenitally Athymic (Nude) Mice.J Exp Med 1973;138:488-94. Nazzaro-Porro,et al.Identification of Tyrosinase Inhibitors in Cultures of Pityrosporum.The Journal of Investigative Dermatology 1978;71:205-208. Noakes.The Aryl Hydrocarbon Receptor:A Review of Its Role in the Physiology and Pathology of the Integument and Its Relationship to the Tryptophan Metabolism.Journal of Tryptophan Research 2015;8:17-18. Otulakowski,et al.Use of a Human Skin-Grafted Nude Mouse Model for the Evaluation of Topical Retinoic Acid Treatment.J Invest Dermatol 1994;102:515-8. Park,J.I.,Lee,H.Y.,Lee,J.E.,Myung,C.H.,Hwang,J.S.Inhibitory effect of 2-methyl-naphtho[1,2,3-de]quinolin-8-one on melanosome transport and skin pigmentation.Sci.Rep.Jul.6:6:29189.Doi:10.1038 / srep29189(2016). Plenat,et al.Host-Donor Interactions in Healing of Human Split-Thickness Skin Grafts Onto Nude Mice:In Situ Hybridization,Immunohistochemical and Histochemical Studies.Transplantation 1992;53:1002-10.Reed,et al.Long-Term Maintenance of Normal Human Skin on Congenitally Athymic(Nude) Mice.Proc Soc Exp Biol Med 1973;143:350-3. Scott,et al.The Permeability of Grafted Human Transplant Skin in Athymic Mice.J Pharm Pharmacol 1988;40:128-9. Song,et al.A Ligand For The Aryl Hydrocarbon Receptor Isolated From Lung.PNAS 2002;99(23):14694-9. Taylor,et al.The Taylor Hyperpigmentation Scale:a new visual assessment tool for the evaluation of skin color and pigmentation.Cutis.2005 Oct;76(4):270-4. Wang,et al.Stress-Induced RNASET2 Overexpression Mediates Melanocyte Apoptosis Via The TRAF2 Pathway In Vitro.Cell Death and Disease 2014;5:e1022 Wasmeier,et al.Melanosomes At A Glance.Journal of Cell Science 2008;121:3995-3999. Wille,et al.Malassezin-A Novel Agonist of the Arylhydrocarbon Receptor From The Yeast Malassezia furfur.Bioorganic&Medicinal Chemistry 2001;9:955-60. Winston-McPherson,et al.Synthesis and Biological Evaluation of 2,3’-diindolylmethanes as Agonists of Aryl Hydrocarbon Receptor.Bioorganic&Medicinal Chemistry Letters 2014;24:4023-4025. Whyte,et al.Ethoxyresorufin-O-deethylase(EROD) Activity in Fish As A Biomarker of Chemical Exposure.Critical Reviews in Toxicology 2000;30(4):347-570. Yamaguchi,et al.Melanocytes and Their Diseases.Cold Spring Harb Perspect Med 2014;4:a017046. Zonios,et al.Skin Melanin,Hemoglobin,and Light Scattering Properties can be Quantitatively Assessed In Vivo Using Diffuse Reflectance Spectroscopy.J Invest Dermatol.2001;117:1452-1457.

[0302] All documents cited in this application are hereby incorporated by reference as if fully set forth herein.

[0303] Although illustrative embodiments of the present invention have been described herein, it should be understood that the present invention is not limited to those described embodiments, and that various other changes or modifications may be made by those skilled in the art without departing from the scope or spirit of the present invention.

[0304] In certain embodiments, for example, the following are provided: (Item 1) Formula (II): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl; At least one of R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 is methyl. or a compound having the structure of a crystal form, a hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof. (Item 2) [ka] and [ka] The compound according to item 1, selected from the group consisting of: (Item 3) Formula (III): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl; At least one of R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 is methyl. or a compound having the structure of a crystalline form, a hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof. (Item 4) The compound according to item 3, which has the following formula: [ka] (Item 5) Formula (II): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl. or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof, A compound for whitening the skin. (Item 6) [ka] , [ka] and [ka] 6. The compound according to item 5, selected from the group consisting of: (Item 7) Formula (III): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl. or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof, A compound for whitening the skin. (Item 8) [ka] and [ka] 8. The compound according to item 7, selected from the group consisting of: (Item 9) Formula (II): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl. or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof, Compounds for inducing apoptosis of melanocytes. (Item 10) [ka] , [ka] and [ka] 10. The compound according to item 9, selected from the group consisting of: (Item 11) Formula (III): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl. or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof, Compounds for inducing apoptosis of melanocytes. (Item 12) [ka] and [ka] 12. The compound according to item 11, selected from the group consisting of: (Item 13) Formula (II): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl. or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof, A compound for agonizing the aryl hydrocarbon receptor (AhR). (Item 14) [ka] , [ka] and [ka] Item 14. The compound according to item 13, selected from the group consisting of: (Item 15) Formula (III): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl. or a crystalline form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof, A compound for agonizing the aryl hydrocarbon receptor (AhR). (Item 16) [ka] and [ka] 16. The compound according to item 15, selected from the group consisting of: (Item 17) Formula (II): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl. or a crystal form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof; and a cosmetically or pharma- ceutically acceptable vehicle, diluent, or carrier. (Item 18) The compound is [ka] , [ka] and [ka] Item 18. The composition according to item 17, selected from the group consisting of: (Item 19) Formula (III): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl. or a crystal form, hydrate, or cosmetically or pharma- ceutically acceptable salt thereof; and a cosmetically or pharma- ceutically acceptable vehicle, diluent, or carrier. (Item 20) The compound is [ka] and [ka] 20. The composition according to item 19, selected from the group consisting of: (Item 21) 1. A method for whitening the skin in a subject, comprising administering to a subject a compound of formula (II): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl. or a crystal form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, to said subject. (Item 22) The compound is [ka] , [ka] and [ka] 22. The method according to item 21, wherein the compound is selected from the group consisting of: (Item 23) 1. A method for whitening the skin in a subject, comprising administering to a subject a compound of formula (III): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl. or a crystal form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, to said subject. (Item 24) The compound is [ka] and [ka] 24. The method according to item 23, wherein the compound is selected from the group consisting of: (Item 25) A method for inducing apoptosis of melanocytes in a subject, comprising administering to a subject an antibody of formula (II): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl. or a crystal form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, to said subject. (Item 26) The compound is [ka] , [ka] and [ka] 26. The method of claim 25, wherein the compound is selected from the group consisting of: (Item 27) A method for inducing apoptosis of melanocytes in a subject, comprising administering to a subject a compound of formula (III): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl. or a crystal form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, to said subject. (Item 28) The compound is [ka] and [ka] 28. The method according to item 27, wherein the compound is selected from the group consisting of: (Item 29) A method for agonizing an aryl hydrocarbon receptor (AhR) in a subject, comprising administering to a subject a compound of formula (II): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, and R11 are independently selected from the group consisting of hydrogen and methyl. or a crystal form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, to said subject. (Item 30) The compound is [ka] , [ka] and [ka] 30. The method of claim 29, wherein the compound is selected from the group consisting of: (Item 31) A method for agonizing an aryl hydrocarbon receptor (AhR) in a subject, comprising administering to a subject a compound of formula (III): [ka] (In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 are independently selected from the group consisting of hydrogen and methyl. or a crystal form, hydrate, or a cosmetically or pharma- ceutically acceptable salt thereof, to said subject. (Item 32) The compound is [ka] and [ka] 32. The method according to claim 31, wherein the compound is selected from the group consisting of:

Claims

[Claim 1] A higher level of safety.

Images