US20180273482A1 - Compositions and methods for increasing, or preventing or reversing loss of, skin pigmentation in a mammalian subject - Google Patents

Compositions and methods for increasing, or preventing or reversing loss of, skin pigmentation in a mammalian subject Download PDF

Info

Publication number
US20180273482A1
US20180273482A1 US15/562,776 US201615562776A US2018273482A1 US 20180273482 A1 US20180273482 A1 US 20180273482A1 US 201615562776 A US201615562776 A US 201615562776A US 2018273482 A1 US2018273482 A1 US 2018273482A1
Authority
US
United States
Prior art keywords
group
alkyl
compound
och
bond
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US15/562,776
Other languages
English (en)
Inventor
Todd RIDKY
Chris NATALE
Jeffrey Winkler
Rosa Ellen COOKSON
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Pennsylvania Penn
Original Assignee
University of Pennsylvania Penn
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University of Pennsylvania Penn filed Critical University of Pennsylvania Penn
Priority to US15/562,776 priority Critical patent/US20180273482A1/en
Assigned to NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT reassignment NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT CONFIRMATORY LICENSE (SEE DOCUMENT FOR DETAILS). Assignors: UNIVERSITY OF PENNSYLVANIA
Publication of US20180273482A1 publication Critical patent/US20180273482A1/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61G—TRANSPORT, PERSONAL CONVEYANCES, OR ACCOMMODATION SPECIALLY ADAPTED FOR PATIENTS OR DISABLED PERSONS; OPERATING TABLES OR CHAIRS; CHAIRS FOR DENTISTRY; FUNERAL DEVICES
    • A61G17/00—Coffins; Funeral wrappings; Funeral urns
    • A61G17/04—Fittings for coffins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/357—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having two or more oxygen atoms in the same ring, e.g. crown ethers, guanadrel
    • A61K31/36—Compounds containing methylenedioxyphenyl groups, e.g. sesamin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/473—Quinolines; Isoquinolines ortho- or peri-condensed with carbocyclic ring systems, e.g. acridines, phenanthridines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/14—Drugs for dermatological disorders for baldness or alopecia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/16—Emollients or protectives, e.g. against radiation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/18—Antioxidants, e.g. antiradicals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D221/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00
    • C07D221/02—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00 condensed with carbocyclic rings or ring systems
    • C07D221/04—Ortho- or peri-condensed ring systems
    • C07D221/06—Ring systems of three rings
    • C07D221/16—Ring systems of three rings containing carbocyclic rings other than six-membered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C07D491/044—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C07D491/048—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring the oxygen-containing ring being five-membered

Definitions

  • Melanin controls the amount of UV radiation from the sun that penetrates the skin by absorption. While UV radiation can assist in the production of vitamin D, excessive exposure to UV can damage health. Melanin is produced within the skin in cells called melanocytes, and is the main determinant of the skin color of darker-skinned humans. Both the amount and type of melanin produced is controlled by a number of genes that operate under incomplete dominance. Melanocytes residing in the basal epidermis control human skin color through synthesis of melanin (melanogenesis), a complex process thought to be primarily regulated by alpha-melanocyte stimulating hormone ( ⁇ MSH).
  • ⁇ MSH alpha-melanocyte stimulating hormone
  • the ⁇ MSH peptide is secreted centrally by the pituitary, and locally by surrounding keratinocytes in response to UV damage.
  • M1R melanocortin receptor 1
  • GPCR G protein-coupled receptor
  • This secondary messenger activates a cascade of downstream transcriptional events leading to expression of genes required for melanin synthesis.
  • melanosomes Melanogenesis takes place within small membrane-bound packages called melanosomes. As the melanosomes become full of melanin, they move into the slender arms of melanocytes, from where they are transferred to the keratinocytes. Under normal conditions, melanosomes cover the upper part of the keratinocytes and protect them from genetic damage.
  • melanocyte supplies melanin to about 36 keranocytes according to signals from the keranocytes. They also regulate melanin production and replication of melanocytes. People have different skin colors mainly because their melanocytes produce different amount and kinds of melanin.
  • the genetic mechanism behind human skin color is mainly regulated by the enzyme tyrosinase, which creates the color of the skin, eyes, and hair shades. Differences in skin color are also attributed to differences in size and distribution of melanosomes in the skin.
  • Uneven pigmentation of some sort affects most people, regardless of bioethnic background or skin color. Skin may appear lighter or darker than normal, or lack pigmentation at all. There may be blotchy and uneven areas, patches of brown to gray discoloration, or freckling. Apart from blood-related conditions such as jaundice, carotenosis, or argyria, skin pigmentation disorders generally occur because the body produces either too much or too little melanin.
  • Increased melanin production can be caused by distinct phenomena, such as melasma (darkening of the skin), chloasma (skin discolorations caused by hormones, which are usually associated with pregnancy, birth control pills or estrogen replacement therapy), and solar lentigo (also known as “liver spots” or “senile freckles,” which are darkened spots on the skin caused by aging and the sun). Liver spots are quite common in adults with a long history of unprotected sun exposure. Aside from sun exposure and hormones, hyperpigmentation can be caused by skin damage, such as remnants of blemishes, wounds or rashes.
  • compositions and methods for increasing, or preventing or reversing loss of, skin pigmentation in a mammalian subject, such as a human Such compositions and methods would be useful, in non-limiting embodiments, for treating or preventing skin disorders or diseases associated with hypopigmentation.
  • the present invention fulfills this need.
  • the invention provides a compound, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof.
  • the invention further provides a pharmaceutical composition comprising at least one pharmaceutically acceptable carrier and at least one compound of the invention, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof.
  • the invention further provides a method of increasing, or preventing or reversing loss of, skin pigmentation in a mammalian subject.
  • the invention further provides a kit for increasing, or preventing or reversing loss of, skin pigmentation in a mammalian subject.
  • the compound of the invention is selected from the group consisting of:
  • R 1 is selected from the group consisting of ⁇ O, ⁇ N—OH, ⁇ N—NHC( ⁇ O)(p-methoxy phenyl), ⁇ N—NHC( ⁇ O)CH(OMe)phenyl, and ⁇ N—NH(5-iodo-pyrid-2-yl);
  • R 2 is selected from the group consisting of C 1 -C 4 alkyl and C 1 -C 4 haloalkyl:
  • bond a is a single or double bond, such that: if bond a is a double bond, R 3 and R 4 are H, and if bond a is a single bond, R 3 is selected from the group consisting of H, —OH, —OAc, and halo; R 4 is selected from the group consisting of H, —OH, —OAc, and —S(o-nitrophenyl); or R 3 and R 4 combine to form a diradical selected from the group consisting of —CH 2 —, —OCH 2 O—, —OCH(CH 3 )O—, and —OC(CH 3 ) 2 O—;
  • R 5 is selected from the group consisting of H, benzyl, C 1 -C 4 alkyl, and acetyl;
  • R 6 is selected from the group consisting of H, halo, —NO 2 , C 1 -C 4 alkyl, —C ⁇ CH, —C ⁇ C—Si(CH 3 ) 3 (or —C ⁇ C-TMS), —O-benzyl, —OH, —OAc, C 1 -C 4 alkoxy, —COOH, and —COO(C 1 -C 4 alkyl);
  • R 7 is selected from the group consisting of H, halo, —NO 2 , C 1 -C 4 alkyl, —OH, —OAc, and C 1 -C 4 alkoxy;
  • R 8 is selected from the group consisting of H, halo, —NO 2 , C 1 -C 4 alkyl, —O-benzyl, —N(R)(R), —SR, —COOH, —COO(C 1 -C 4 alkyl), —OH, —OAc, C 1 -C 4 alkoxy, 3-thietyl-methoxy, —SO 2 (morpholino), and —OCH 2 CH ⁇ CH 2 , wherein each occurrence of R is independently selected from the group consisting of H and C 1 -C 4 alkyl;
  • R 9 is selected from the group consisting of H, halo, —NO 2 , C 1 -C 4 alkyl, —OH, —OAc, and C 1 -C 4 alkoxy,
  • R 10 is selected from the group consisting of H, C 1 -C 4 alkyl, and halo;
  • each occurrence of benzyl is independently optionally substituted with at least one group selected from the group consisting of C 1 -C 4 alkyl, —OH, C 1 -C 4 alkoxy, halo, and —NO 2 ; and a molecule of formula (II):
  • R 1 is selected from the group consisting of ⁇ O and ⁇ N—OH;
  • R 2 is C 1 -C 4 alkyl
  • R 5 is selected from the group consisting of H, benzyl and C 1 -C 4 alkyl:
  • R 8 and R 9 are independently selected from the group consisting of H and C 1 -C 4 alkoxy, or R 8 and R 9 combine to form a diradical selected from the group consisting of —OCH 2 O—, —OCH(CH 3 )O— and —OC(CH 3 ) 2 O—.
  • the compound is not G-1 (or G1; rel-1-[4-(6-bromo-1,3-benzodioxol-5-yl)-3aR,4S,5,9bS-tetrahydro-3H-cyclopenta[c]quinolin-8-yl]-ethanone):
  • the compound is G-1.
  • the compound is not selected from the group consisting of A1-A107. In other embodiments, the compound is at least one selected from the group consisting of A1-A107. In yet other embodiments, the compound is not selected from the group consisting of CMPD1-CMPD12. In yet other embodiments, the compound is selected from the group consisting of CMPD1-CMPD12.
  • the compound, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof is selected from the group consisting of:
  • R 1 is selected from the group consisting of ⁇ O and ⁇ N—OH;
  • R 2 is C 1 -C 4 alkyl
  • bond a is a single or double bond, such that: if bond a is a double bond, R 3 and R 4 are H, and if bond a is a single bond, R 3 and R 4 are independently selected from the group consisting of H and —OH, or R 3 and R 4 combine to form a diradical selected from the group consisting of —OCH 2 O—, —OCH(CH 3 )O— and —OC(CH 3 ) 2 O—;
  • R 5 is selected from the group consisting of H, benzyl and C 1 -C 4 alkyl;
  • R 6 is selected from the group consisting of H and halo
  • R 8 and R 9 are independently selected from the group consisting of H and C 1 -C 4 alkoxy, or R 8 and R 9 combine to form a diradical selected from the group consisting of —OCH 2 O—, —OCH(CH 3 )O— and —OC(CH 3 ) 2 O—;
  • R 1 is selected from the group consisting of ⁇ O and ⁇ N—OH;
  • R 2 is C 1 -C 4 alkyl
  • R 5 is selected from the group consisting of H, benzyl and C 1 -C 4 alkyl;
  • R 8 and R 9 are independently selected from the group consisting of H and C 1 -C 4 alkoxy, or R 8 and R 9 combine to form a diradical selected from the group consisting of —OCH 2 O—, —OCH(CH 3 )O— and —OC(CH 3 ) 2 O—.
  • the compound is not G-1.
  • R 1 is ⁇ O.
  • R 1 is ⁇ N—OH.
  • R 2 is selected from the group consisting of methyl, trifluoromethyl, ethyl, 1-propyl, and 2-propyl.
  • bond a is a double bond
  • R 3 and R 4 are H.
  • bond a is a single bond
  • R 3 and R 4 are independently selected from the group consisting of H and —OH.
  • bond a is a single bond, and R 3 and R 4 are H.
  • bond a is a single bond, and R 3 and R 4 are —OH.
  • R 3 and R 4 are —OH and cis to each other.
  • bond a is a single bond, and R 3 and R 4 combine to form —OC(CH 3 ) 2 O—.
  • R 5 is selected from the group consisting of H, benzyl, methyl, ethyl, 1-propyl and 2-propyl.
  • R 6 is selected from the group consisting of H, F, Cl, Br and I.
  • R 8 and R 9 are independently selected from the group consisting of H, methoxy, ethoxy, 1-propoxy and 2-propoxy.
  • R 8 and R 9 combine to form a diradical selected from the group consisting of —OCH 2 O—, —OCH(CH 3 )O— and —OC(CH 3 ) 2 O—.
  • (1) is at least one selected from the group consisting of:
  • the compound is at least one selected from the group consisting of: CMPD1 (rel-1-((3aS,4R,9bR)-4-(benzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one); CMPD2 (rel-1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-5-methyl-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one); CMPD3 (rel-1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-2,3,3a,4,5,9b-hexa
  • CMPD4 (rel-1-((3aS,4R,9bR)-5-benzyl-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one);
  • CMPD5 (rel-1-((3aS,4R,9bR)-4-(2-bromophenyl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one);
  • CMPD6 (rel-1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one oxi
  • the pharmaceutical composition is formulated for topical or transdermal administration.
  • the pharmaceutical composition does not comprise G-1. In other embodiments, the pharmaceutical composition comprises at least one compound selected from the group consisting of G-1, CMPD1-CMPD12, and A1-A107. In yet other embodiments, the pharmaceutical composition comprises at least one compound selected from the group consisting of CMPD1-CMPD12. In other embodiments, the pharmaceutical composition comprises at least one compound selected from the group consisting of G-11 and CMPD1-CMPD12.
  • the pharmaceutical composition further comprises at least one sun-blocking agent. In certain embodiments, the pharmaceutical composition further comprises at least sunscreen lotion.
  • the method comprises administering to the subject a therapeutically effective amount of a compound, wherein the compound is a GPER agonist and does not bind to a canonical nuclear estrogen receptor.
  • the subject is human.
  • the canonical nuclear estrogen receptor comprises at least one selected from the group consisting of ER ⁇ and ER ⁇ .
  • the compound is a GPER1 or GPR30 agonist.
  • the subject is suffering from at least one condition selected from the group consisting of pigmentary changes associated with oral contraceptive use, pregnancy, and endogenous estrogens in females; solar lentigo; acne; eczema; chemical, sun, and thermal burn scars; and lupus.
  • the compound is administered topically or transdermally to the subject.
  • the compound is formulated as a pharmaceutical composition for topical or transdermal administration.
  • the pharmaceutical composition further comprises at least one sun-blocking agent.
  • the pharmaceutical composition comprises a sunscreen lotion.
  • the pharmaceutical composition is essentially free of a skin bleaching agent.
  • the subject is not further submitted to UV radiation as part of the treatment to increase, or prevent or reverse loss of, skin pigmentation.
  • the compound is at least one selected from the group consisting of G1, A1-A107 and CMPD1-CMPD12.
  • the kit comprises at least one compound or pharmaceutical composition of the invention. In other embodiments, the kit comprises instructions for topically or transdermally administering the at least one pharmaceutical composition to the subject.
  • FIG. 1 comprises a set of images illustrating pregnancy-associated pigmentation disorder.
  • Images A-C depict pigmentation disorder of patients with melasma.
  • Image D depicts a patient with pigmentary demarcation lines.
  • Images E and F depict typical linea nigra on a mother.
  • Image G depicts typical linea nigra on a child.
  • FIGS. 2A-2F comprise a set of scheme and bar graphs illustrating melanin production in human melanocytes.
  • FIG. 2A comprises a schematic illustration of classical regulation of human melanocyte homeostasis.
  • FIG. 2B comprises a bar graph illustrating melanin production in response to ⁇ MSH in a dose dependent manner.
  • FIG. 2C comprises a bar graph illustrating melanin production in response to estrogen in a dose dependent manner.
  • FIG. 2D comprises a bar graph illustrating melanin production by melanocytes treated with vehicle, ⁇ MSH, or estrogen.
  • FIG. 2E comprises a bar graph illustrating melanin production in response to estrogen using iPS-derived female melanocytes.
  • FIGS. 3A-3C comprise a set of images and bar graphs illustrating the finding that estrogen increases melanin synthesis.
  • FIG. 3A comprises a bar graph illustrating melanin content of primary human melanocytes treated with estrogen (E2), compared to vehicle-treated controls.
  • FIG. 3B comprises a bar graph and images illustrating Fontana-Masson (melanin) staining of organotypic skin treated with vehicle or estrogen. Relative melanin content is illustrated in the bar graph at the bottom of the Figure.
  • FIGS. 4A-4D comprise a set of images and bar graphs illustrating the finding that estrogen accesses the classical melanin production pathway through nonclassical receptors.
  • FIG. 4A comprises a bar graph illustrating cAMP ELISA from estrogen-treated melanocytes.
  • FIG. 4B comprises a set of western blot images demonstrating changes in classical melanin pathway regulators after estrogen treatment.
  • FIG. 4C comprises a set of western blot images for estrogen receptors in MCF7 cells and melanocytes.
  • FIGS. 5A-5F comprises a set of bar graphs and table illustrating hormone receptors in melanocytes.
  • FIG. 5A comprises a bar graph illustrating relative gene expression of classical hormone receptors in MCF7 cells and melanocytes, as determined by qRT-PCR. Ct values were normalized to actin, and set relative to the expression of androgen receptor (AR) in MCF7 cells.
  • FIG. 5B comprises a table illustrating RPKM values for classical and nonclassical estrogen receptors in human melanocytes, by convention, RPKM values >1 indicate the gene is expressed.
  • FIG. 5C comprises a bar graph illustrating expression of GPER displayed as 1/Ct value.
  • 5D comprises a bar graph illustrating relative expression of transcripts in melanocytes, fibroblasts, and keratinocytes, as determined by qRT-PCR, displayed relative to the expression level in melanocytes.
  • FIG. 5E comprises a bar graph illustrating qRT-PCR showing mRNA knockdown efficiency of the two hairpins targeting GPER.
  • FIG. 5F comprises a set of bar graphs illustrating melanin content of melanocytes transduced with LentiCRISPRV2 with guide RNA targeting GFP or GPER. Cells were treated with either vehicle or estrogen. Error bars denote ⁇ s.d., *p ⁇ 0.05.
  • FIGS. 6A-6B comprise a set of bar graphs illustrating the finding that melanin production is altered by sex steroid analogs-GPER agonists currently in clinical use.
  • FIG. 6A is a bar graph illustrating melanin production in response to tamoxifen (TMX).
  • FIG. 6B is a bar graph illustrating melanin production in response to ethinyl estradiol (EE2).
  • N 3 biologic replicates for each experiment. Error bars denote ⁇ s.d., *p ⁇ 0.05.
  • FIGS. 7A-7B comprise a set of bar graphs illustrating the effects of specific GPER agonists on melanin production.
  • FIG. 7A comprises a bar graph illustrating melanin production in response to G-1, a specific GPER agonist.
  • FIG. 7B comprises a bar graph illustrating melanin assay performed on melanocytes lentivirally transduced with control shRNA or shRNA targeting GPER. These cells were treated with either vehicle or G-1.
  • FIGS. 8A-8C comprise a set of images and bar graphs illustrating the finding that GPER signaling is sufficient to alter melanin production in organotypic human tissue.
  • FIG. 8A comprises an image illustrating organotypic skin treated with vehicle (left) or G-1 (right).
  • FIG. 8B comprises images and a bar graph illustrating Fontana-Masson (melanin) staining of organotypic skin treated with vehicle or G-1. Quantification of melanin content is shown on the right.
  • FIGS. 9A-9D comprise a set of bar graphs and images illustrating the finding that topical GPER agonists increase pigmentation in vivo.
  • FIG. 9A comprises an image illustrating mouse ear skin treated for 3 weeks with vehicle only on the left ear, and 2% G-1 on the right ear.
  • FIG. 9B comprises a bar graph illustrating melanin assay on whole ear tissue that was treated with either vehicle or 2% G-1 for 3 weeks.
  • FIG. 9C comprises a set of images illustrating Fontana-Masson (melanin) staining of tissue sections from ears treated with either vehicle or 2% G-1, quantification of staining on right.
  • FIG. 10 comprises a set of western blot images illustrating the finding that compounds of the invention increase levels of pCREB in human primary melanocytes.
  • the actin control is illustrated at the bottom of the images.
  • the present invention relates, in one aspect, to the unexpected discovery that compounds of the invention modulate skin pigmentation in a mammalian subject, such as a human.
  • the compounds of the invention treat or prevent skin hypopigmentation in a subject.
  • the compounds of the invention treat or prevent wrinkles and/or discolorations of premature aging.
  • the compositions of the invention modulate skin color for esthetic benefit without exposure to DNA-damaging UV radiation.
  • the canonical nuclear ER comprises ER ⁇ and/or ER ⁇ .
  • GPER agonists such as but not limited to G-1
  • G-1 represent a useful, novel class of therapeutics for normalizing disorders of epidermal pigmentation.
  • Myriad genetic and acquired conditions including common afflictions such as acne, eczema, vitiligo, ultraviolet (UV) radiation exposure, traumatic injury, and pregnancy are associated with alterations in skin pigmentation that can be extensive and long-lasting.
  • a population that could potentially benefit from modulating skin pigment are people with naturally light skin, especially those with red hair, who have a markedly decreased ability to synthesize UV-protective brown eumelanin as a result of inactivating mutations in MC1R.
  • This large population is especially susceptible to photodamage, sunburns, and has an increased lifetime risk of keratinocyte and melanocyte-derived skin cancers.
  • the only method currently available to increase skin melanin is UV exposure, which is effective at darkening skin but promotes DNA damage, which leads to premature aging, wrinkles, and skin cancer.
  • the specific activation of GPER alternatively activates cAMP signaling, bypassing MC1R, to stimulate melanin synthesis, and can thus be especially useful in this sun-vulnerable population.
  • Selective GPER activation in skin can potentially be a safe alternative to intentional UV radiation exposure (via natural sunlight or tanning beds) for individuals seeking what they perceive as an esthetically desirable tan.
  • targeting the receptor(s) described herein can protect skin from wrinkles, and discolorations of premature aging, and also modulate skin color for esthetic benefit without exposure to DNA-damaging UV radiation or potentially toxic skin bleaching agents.
  • the invention should not be construed to be limited to the estrogen derivatives, or any analog thereof, recited herein.
  • the present invention encompasses any estrogen derivatives, or analogs thereof, that act as agonists towards GPER and do not bind to the canonical nuclear estrogen receptor.
  • the articles “a” and “an” refer to one or to more than one (i.e., to at least one) of the grammatical object of the article.
  • an element means one element or more than one element.
  • ⁇ MSH alpha-melanocyte stimulating hormone
  • the term “about” will be understood by persons of ordinary skill in the art and will vary to some extent on the context in which it is used. As used herein when referring to a measurable value such as an amount, a temporal duration, and the like, the term “about” is meant to encompass variations of ⁇ 20% or ⁇ 10%, ⁇ 5%, ⁇ 1%, or ⁇ 0.1% from the specified value, as such variations are appropriate to perform the disclosed methods.
  • alkoxy employed alone or in combination with other terms means, unless otherwise stated, an alkyl group having the designated number of carbon atoms, as defined elsewhere herein, connected to the rest of the molecule via an oxygen atom, such as, for example, methoxy, ethoxy, 1-propoxy, 2-propoxy (or isopropoxy) and the higher homologs and isomers.
  • oxygen atom such as, for example, methoxy, ethoxy, 1-propoxy, 2-propoxy (or isopropoxy) and the higher homologs and isomers.
  • a specific example is (C 1 -C 3 )alkoxy, such as, but not limited to, ethoxy and methoxy.
  • alkyl by itself or as part of another substituent means, unless otherwise stated, a straight or branched chain hydrocarbon having the number of carbon atoms designated (i.e., C 1 -C 10 means one to ten carbon atoms) and includes straight, branched chain, or cyclic substituent groups. Examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl, hexyl, and cyclopropylmethyl.
  • a specific embodiment is (C 1 -C 4 )alkyl, such as, but not limited to, ethyl, methyl, isopropyl, n-butyl, isobutyl, t-butyl, and cyclopropylmethyl.
  • AR refers to androgen receptor
  • cAMP refers to cyclic adenosine monophosphate
  • CREB refers to cAMP response element-binding protein
  • a “disease” is a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated, the animal's health continues to deteriorate.
  • a “disorder” in an animal is a state of health in which the animal is able to maintain homeostasis, but in which the animal's state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a further decrease in the animal's state of health.
  • E2 refers to estrogen
  • EE2 refers to ethinyl estradiol.
  • the terms “effective amount” or “therapeutically effective amount” or “pharmaceutically effective amount” of a compound are used interchangeably to refer to the amount of the compound sufficient to provide a beneficial effect to the subject to which the compound is administered.
  • ER refers to estrogen receptor
  • G1 or “G-1” refers to rel-1-[4-(6-bromo-1,3-benzodioxol-5-yl)-3aR,4S,5,9bS-tetrahydro-3H-cyclopenta[c]quinolin-8-yl]-ethanone), or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof.
  • GPCR refers to a G protein-coupled receptor
  • GPR30 receptor also known as GPER1; CEPR; CMKRL2; DRY12; FEG-1; GPCR-Br; GPER; LERGU; LERGU2; LyGPR; or mER
  • 7TM 7-transmembrane 7TM G protein-coupled receptor that mediate estrogen-dependent signal transduction.
  • GPR30 is an intracellular protein, found in the endoplasmic reticulum, which binds estrogen with high affinity (K d of 6 nM) and mediates rapid cellular responses including calcium mobilization and phosphatidylinositol 3,4,5-trisphosphate production in the nucleus.
  • GPR30 receptor refers to all types of GPR30 receptor, regardless of the tissue in which such receptor is found and refers to any variant thereof, including receptors of mammals (such as humans and domesticated mammals where veterinary applications are relevant) and variants thereof.
  • Other names which have been used for GPR30 include CMKRL2, DRY12, FEG-1. GPCR-Br, LERGU, LERGU2, LyGPR. CEPR and MGC99678, among others.
  • halo or “halogen” alone or as part of another substituent refers to, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.
  • an “instructional material” includes a publication, a recording, a diagram, or any other medium of expression that can be used to communicate the usefulness of a compound, composition, assay or method of the invention in a kit for suppressing or reducing systemic immune response in a subject.
  • the instructional material of the kit of the invention can, for example, be affixed to a container which contains the identified compound, composition, assay, or methods of the invention or be shipped together with a container that contains the identified compound, composition, assay, or method.
  • the instructional material can be shipped separately from the container with the intention that the instructional material and the compound, composition, assay, or method be used cooperatively by the recipient.
  • M1R melanocortin receptor 1.
  • MTF microphthalmia-associated transcription factor
  • modulate means, with respect to disease states or conditions associated with binding of a compound of the present invention to a receptor contemplated in the present invention, to produce, either directly or indirectly, an improvement or lessening of a condition or disease state which was, prior to administration of a compound according to the present invention, sub-optimal and in many cases, debilitating and even life threatening. Modulation may occur by virtue of agonist activity, antagonist activity or mixed agonist/antagonist activity (depending on the receptor site).
  • Parenteral administration of a composition includes, e.g., subcutaneous (s.c.), intravenous (i.v.), intramuscular (i.m.), or intrasternal injection, or infusion techniques.
  • composition refers to a mixture of at least one compound useful within the invention with other chemical components, such as carriers, stabilizers, diluents, dispersing agents, suspending agents, thickening agents, and/or excipients.
  • the pharmaceutical composition facilitates administration of the compound to an organism. Multiple techniques of administering a compound exist in the art including, but not limited to: intravenous, oral, aerosol, parenteral, ophthalmic, pulmonary, intracranial, transdermal and topical administration. In certain embodiments, the administration comprises topical administration.
  • the term “pharmaceutically acceptable” refers to a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the composition, and is relatively non-toxic, i.e., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.
  • the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound useful within the invention within or to the subject such that it may perform its intended function.
  • a pharmaceutically acceptable material, composition or carrier such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound useful within the invention within or to the subject such that it may perform its intended function.
  • Such constructs are carried or transported from one organ, or portion of the body, to another organ, or portion of the body.
  • Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, including the compound useful within the invention, and not injurious to the subject.
  • materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic saline
  • “pharmaceutically acceptable carrier” also includes any and all coatings, antibacterial and antifungal agents, and absorption delaying agents, and the like that are compatible with the activity of the compound useful within the invention, and are physiologically acceptable to the subject. Supplementary active compounds may also be incorporated into the compositions.
  • the “pharmaceutically acceptable carrier” may further include a pharmaceutically acceptable salt of the compound useful within the invention.
  • Other additional ingredients that may be included in the pharmaceutical compositions used in the practice of the invention are known in the art and described, for example in Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, Pa.), which is incorporated herein by reference.
  • pharmaceutically acceptable salt refers to a salt of the administered compound prepared from pharmaceutically acceptable non-toxic acids and/or bases, including inorganic acids, inorganic bases, organic acids, inorganic bases, solvates (including hydrates) and clathrates thereof.
  • a “pharmaceutically effective amount,” “therapeutically effective amount” or “effective amount” of a compound is that amount of compound that is sufficient to provide a beneficial effect to the subject to which the compound is administered.
  • prevent means avoiding or delaying the onset of symptoms associated with a disease or condition in a subject that has not developed such symptoms at the time the administering of an agent or compound commences.
  • Disease, condition and disorder are used interchangeably herein.
  • the terms “subject” and “individual” and “patient” can be used interchangeably and may refer to a human or non-human mammal or a bird.
  • Non-human mammals include, for example, livestock and pets, such as ovine, bovine, porcine, canine, feline and murine mammals.
  • the subject is human.
  • TMX refers to tamoxifen
  • topical administration or “topical application” refers to a medication applied to body surfaces such as the skin or mucous membranes.
  • treat means reducing the frequency with which symptoms are experienced by a patient or subject or administering an agent or compound to reduce the severity with which symptoms are experienced.
  • An appropriate therapeutic amount in any individual case may be determined by one of ordinary skill in the art using routine experimentation.
  • treatment is defined as the application or administration of a therapeutic agent, i.e., a composition useful within the invention (alone or in combination with another pharmaceutical agent), to a subject, or application or administration of a therapeutic agent to an isolated tissue or cell line from a subject (e.g., for diagnosis or ex vivo applications), who has a disease or disorder, a symptom of a disease or disorder or the potential to develop a disease or disorder, with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve or affect the disease or disorder, the symptoms of the disease or disorder or the potential to develop the disease or disorder.
  • Such treatments may be specifically tailored or modified, based on knowledge obtained from the field of pharmacogenomics.
  • UV refers to ultraviolet
  • range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.
  • the present invention encompasses an estrogen derivative or analog that binds to GPER, such as but not limited to GPER1, and does not bind to the canonical nuclear estrogen receptor (ER).
  • the estrogen derivative or analog is a GPER agonist.
  • the compound of the invention or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof, is at least one compound of formula (I):
  • R 1 is selected from the group consisting of ⁇ O, ⁇ N—OH, ⁇ N—NHC( ⁇ O)(p-methoxy phenyl), ⁇ N—NHC( ⁇ O)CH(OMe)phenyl, and ⁇ N—NH(5-iodo-pyrid-2-yl);
  • R 2 is selected from the group consisting of C 1 -C 4 alkyl and C 1 -C 4 haloalkyl (such as, but not limited to, —CF 3 );
  • bond a is a single or double bond, such that:
  • R 5 is selected from the group consisting of H, benzyl, C 1 -C 4 alkyl, and acetyl;
  • R 6 is selected from the group consisting of H, halo, —NO 2 , C 1 -C 4 alkyl, —C ⁇ CH, —C ⁇ C—Si(CH 3 ) 3 (or —C ⁇ C-TMS), —O-benzyl, —OH, —OAc, C 1 -C 4 alkoxy, —COOH, and —COO(C 1 -C 4 alkyl);
  • R 7 is selected from the group consisting of H, halo, —NO 2 , C 1 -C 4 alkyl, —OH, —OAc, and C 1 -C 4 alkoxy;
  • R 8 is selected from the group consisting of H, halo, —NO 2 , C 1 -C 4 alkyl, —O-benzyl, —N(R)(R), —SR, —COOH, —COO(C 1 -C 4 alkyl), —OH, —OAc, C 1 -C 4 alkoxy, 3-thietyl-methoxy, —SO 2 (morpholino), and —OCH 2 CH ⁇ CH 2 , wherein each occurrence of R is independently selected from the group consisting of H and C 1 -C 4 alkyl;
  • R 9 is selected from the group consisting of H, halo, —NO 2 , C 1 -C 4 alkyl, —OH, —OAc, and C 1 -C 4 alkoxy,
  • R 10 is selected from the group consisting of H, C 1 -C 4 alkyl, and halo;
  • each occurrence of benzyl is independently optionally substituted with at least one group selected from the group consisting of C 1 -C 4 alkyl, —OH, C 1 -C 4 alkoxy, halo, and —NO 2 .
  • the compound of the invention or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof, is at least one compound of formula (I-1):
  • R 1 is selected from the group consisting of ⁇ O and ⁇ N—OH;
  • R 2 is C 1 -C 4 alkyl
  • bond a is a single or double bond, such that:
  • R 5 is selected from the group consisting of H, benzyl and C 1 -C 4 alkyl;
  • R 6 is selected from the group consisting of H and halo
  • R 8 and R 9 are independently selected from the group consisting of H and C 1 -C 4 alkoxy, or R 8 and R 9 combine to form a diradical selected from the group consisting of —OCH 2 O—, —OCH(CH 3 )O— and —OC(CH 3 ) 2 O—.
  • the compound is not G-1. In other embodiments, the compound is G-1.
  • R 1 is ⁇ O. In other embodiments, R 1 is ⁇ N—OH.
  • R 2 is selected from the group consisting of methyl, trifluoromethyl, ethyl, 1-propyl, and 2-propyl. In other embodiments, R 2 is methyl or trifluoromethyl.
  • bond a is a double bond
  • R 3 and R 4 are H.
  • bond a is a single bond, and R 3 and R 4 are independently selected from the group consisting of H and —OH. In other embodiments, bond a is a single bond, and R 3 and R 4 are H. In yet other embodiments, bond a is a single bond, and R 3 and R 4 are —OH. In yet other embodiments, bond a is a single bond, and R 3 and R 4 are —OH and cis to each other. In yet other embodiments, bond a is a single bond, and R 3 and R 4 are —OH and trans to each other.
  • bond a is a single bond, and R 3 and R 4 combine to form —OC(CH 3 ) 2 O—.
  • R 5 is selected from the group consisting of H, benzyl, methyl, ethyl, 1-propyl and 2-propyl. In certain embodiments, R is selected from the group consisting of H, benzyl and methyl.
  • R 6 is selected from the group consisting of H, F, Cl, Br and I.
  • R 5 is selected from the group consisting of H, Cl and Br.
  • R 8 and R 9 are independently selected from the group consisting of H and C 1 -C 4 alkoxy. In other embodiments, R 8 and R 9 are independently selected from the group consisting of H, methoxy, ethoxy, 1-propoxy and 2-propoxy. In yet other embodiments, R 8 and R 9 are independently selected from the group consisting of H and methoxy.
  • R 8 and R 9 combine to form a diradical selected from the group consisting of —OCH 2 O—, —O(CH 2 ) 2 O—, —OCH(CH 3 )O— and —OC(CH 3 ) 2 O—.
  • the compound, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof is at least one selected from the group consisting of:
  • the compound, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof is at least one selected from the group consisting of:
  • the compound is A1. In other embodiments, the compound is A2. In yet other embodiments, the compound is A3. In yet other embodiments, the compound is A4. In yet other embodiments, the compound is A5. In yet other embodiments, the compound is A6. In yet other embodiments, the compound is A7. In yet other embodiments, the compound is A8. In yet other embodiments, the compound is A9. In yet other embodiments, the compound is A10. In yet other embodiments, the compound is A11. In yet other embodiments, the compound is A12. In yet other embodiments, the compound is A13. In yet other embodiments, the compound is A14. In yet other embodiments, the compound is A15.
  • the compound is not A1. In other embodiments, the compound is not A2. In yet other embodiments, the compound is not A3. In yet other embodiments, the compound is not A4. In yet other embodiments, the compound is not A5. In yet other embodiments, the compound is not A6. In yet other embodiments, the compound is not A7. In yet other embodiments, the compound is not A8. In yet other embodiments, the compound is not A9. In yet other embodiments, the compound is not A10. In yet other embodiments, the compound is not A11. In yet other embodiments, the compound is not A12. In yet other embodiments, the compound is not A13. In yet other embodiments, the compound is not A14. In yet other embodiments, the compound is not A15.
  • the compound, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof is at least one selected from the group consisting of:
  • the compound is A16. In other embodiments, the compound is A17. In yet other embodiments, the compound is A18. In yet other embodiments, the compound is A19. In yet other embodiments, the compound is A20. In yet other embodiments, the compound is A21. In yet other embodiments, the compound is A22. In yet other embodiments, the compound is A23. In yet other embodiments, the compound is A24. In yet other embodiments, the compound is A25. In yet other embodiments, the compound is A26. In yet other embodiments, the compound is A27. In yet other embodiments, the compound is A28. In yet other embodiments, the compound is A29. In yet other embodiments, the compound is A30. In yet other embodiments, the compound is A31.
  • the compound is A32. In yet other embodiments, the compound is A33. In yet other embodiments, the compound is A34. In yet other embodiments, the compound is A35. In yet other embodiments, the compound is A36. In yet other embodiments, the compound is A37. In yet other embodiments, the compound is A38. In yet other embodiments, the compound is A39. In yet other embodiments, the compound is A40. In yet other embodiments, the compound is A41. In yet other embodiments, the compound is A42. In yet other embodiments, the compound is A43. In yet other embodiments, the compound is A44. In yet other embodiments, the compound is A45. In yet other embodiments, the compound is A46. In yet other embodiments, the compound is A47.
  • the compound is A48. In yet other embodiments, the compound is A49. In yet other embodiments, the compound is A50. In yet other embodiments, the compound is A51. In yet other embodiments, the compound is A52. In yet other embodiments, the compound is A53. In yet other embodiments, the compound is A54. In yet other embodiments, the compound is A55. In yet other embodiments, the compound is A56. In yet other embodiments, the compound is A57. In yet other embodiments, the compound is A58. In yet other embodiments, the compound is A59. In yet other embodiments, the compound is A60. In yet other embodiments, the compound is A61. In yet other embodiments, the compound is A62. In yet other embodiments, the compound is A63.
  • the compound is A64. In yet other embodiments, the compound is A65. In yet other embodiments, the compound is A66. In yet other embodiments, the compound is A67. In yet other embodiments, the compound is A68. In yet other embodiments, the compound is A69. In yet other embodiments, the compound is A70. In yet other embodiments, the compound is A71. In yet other embodiments, the compound is A72. In yet other embodiments, the compound is A73. In yet other embodiments, the compound is A74. In yet other embodiments, the compound is A75. In yet other embodiments, the compound is A76. In yet other embodiments, the compound is A77. In yet other embodiments, the compound is A78. In yet other embodiments, the compound is A79.
  • the compound is A80. In yet other embodiments, the compound is A81. In yet other embodiments, the compound is A82. In yet other embodiments, the compound is A83. In yet other embodiments, the compound is A84. In yet other embodiments, the compound is A85. In yet other embodiments, the compound is A86. In yet other embodiments, the compound is A87. In yet other embodiments, the compound is A88. In yet other embodiments, the compound is A89. In yet other embodiments, the compound is A90. In yet other embodiments, the compound is A91. In yet other embodiments, the compound is A92. In yet other embodiments, the compound is A93. In yet other embodiments, the compound is A94. In yet other embodiments, the compound is A95.
  • the compound is A96. In yet other embodiments, the compound is A97. In yet other embodiments, the compound is A98. In yet other embodiments, the compound is A99. In yet other embodiments, the compound is A100. In yet other embodiments, the compound is A101. In yet other embodiments, the compound is A102. In yet other embodiments, the compound is A103. In yet other embodiments, the compound is A104.
  • the compound is not A16. In other embodiments, the compound is not A17. In yet other embodiments, the compound is not A18. In yet other embodiments, the compound is not A19. In yet other embodiments, the compound is not A20. In yet other embodiments, the compound is not A21. In yet other embodiments, the compound is not A22. In yet other embodiments, the compound is not A23. In yet other embodiments, the compound is not A24. In yet other embodiments, the compound is not A25. In yet other embodiments, the compound is not A26. In yet other embodiments, the compound is not A27. In yet other embodiments, the compound is not A28. In yet other embodiments, the compound is not A29.
  • the compound is not A30. In yet other embodiments, the compound is not A31. In yet other embodiments, the compound is not A32. In yet other embodiments, the compound is not A33. In yet other embodiments, the compound is not A34. In yet other embodiments, the compound is not A35. In yet other embodiments, the compound is not A36. In yet other embodiments, the compound is not A37. In yet other embodiments, the compound is not A38. In yet other embodiments, the compound is not A39. In yet other embodiments, the compound is not A40. In yet other embodiments, the compound is not A41. In yet other embodiments, the compound is not A42. In yet other embodiments, the compound is not A43.
  • the compound is not A44. In yet other embodiments, the compound is not A45. In yet other embodiments, the compound is not A46. In yet other embodiments, the compound is not A47. In yet other embodiments, the compound is not A48. In yet other embodiments, the compound is not A49. In yet other embodiments, the compound is not A50. In yet other embodiments, the compound is not A51. In yet other embodiments, the compound is not A52. In yet other embodiments, the compound is not A53. In yet other embodiments, the compound is not A54. In yet other embodiments, the compound is not A55. In yet other embodiments, the compound is not A56. In yet other embodiments, the compound is not A57.
  • the compound is not A58. In yet other embodiments, the compound is not A59. In yet other embodiments, the compound is not A60. In yet other embodiments, the compound is not A61. In yet other embodiments, the compound is not A62. In yet other embodiments, the compound is not A63. In yet other embodiments, the compound is not A64. In yet other embodiments, the compound is not A65. In yet other embodiments, the compound is not A66. In yet other embodiments, the compound is not A67. In yet other embodiments, the compound is not A68. In yet other embodiments, the compound is not A69. In yet other embodiments, the compound is not A70. In yet other embodiments, the compound is not A71.
  • the compound is not A72. In yet other embodiments, the compound is not A73. In yet other embodiments, the compound is not A74. In yet other embodiments, the compound is not A75. In yet other embodiments, the compound is not A76. In yet other embodiments, the compound is not A77. In yet other embodiments, the compound is not A78. In yet other embodiments, the compound is not A79. In yet other embodiments, the compound is not A80. In yet other embodiments, the compound is not A81. In yet other embodiments, the compound is not A82. In yet other embodiments, the compound is not A83. In yet other embodiments, the compound is not A84. In yet other embodiments, the compound is not A85.
  • the compound is not A86. In yet other embodiments, the compound is not A87. In yet other embodiments, the compound is not A88. In yet other embodiments, the compound is not A89. In yet other embodiments, the compound is not A90. In yet other embodiments, the compound is not A91. In yet other embodiments, the compound is not A92. In yet other embodiments, the compound is not A93. In yet other embodiments, the compound is not A94. In yet other embodiments, the compound is not A95. In yet other embodiments, the compound is not A96. In yet other embodiments, the compound is not A97. In yet other embodiments, the compound is not A98. In yet other embodiments, the compound is not A99.
  • the compound is not A100. In yet other embodiments, the compound is not A101. In yet other embodiments, the compound is not A102. In yet other embodiments, the compound is not A103. In yet other embodiments, the compound is not A104.
  • the compound of the invention is at least one compound selected from the group consisting of:
  • the compound is not A105. In other embodiments, the compound is not A106.
  • the compound of the invention or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof, is
  • the compound is not A107.
  • the compound is at least one selected from the group consisting of:
  • the compound of the invention or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof, is at least one compound of formula (II):
  • R 1 is selected from the group consisting of ⁇ O and ⁇ N—OH;
  • R 2 is C 1 -C 4 alkyl
  • R 5 is selected from the group consisting of H, benzyl and C 1 -C 4 alkyl;
  • R 8 and R 9 are independently selected from the group consisting of H and C 1 -C 4 alkoxy, or R 8 and R 9 combine to form a diradical selected from the group consisting of —OCH 2 O—, —OCH(CH 3 )O— and —OC(CH 3 ) 2 O—.
  • the compound of formula (II) is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl
  • the compound useful within the methods of the invention, or a salt, solvate, enantiomer or diastereoisomer thereof is at least one GPER agonist recited in U.S. Patent Application Publications No. US 2008/0167334 and US 2011/0092533, all of which are incorporated herein in their entireties by reference:
  • X is ⁇ N—, O, S, or N—R, with the proviso that when X is N—R and R is a bond, N together with R 1 forms a 5- to 7-membered optionally substituted heterocyclic group;
  • R is a bond, H, —OH, —NO 2 , optionally substituted C 1 -C 6 hydrocarbyl (such as optionally substituted alkyl), optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted —C(O)—(C 1 -C 6 ) alkyl (amide), optionally substituted —C(O)—O—(C 1 -C 6 ) alkyl (urethane), optionally substituted —C(O)—NH(C 1 -C 6 ) alkyl (urea), optionally substituted —C(O)—N(C 1 -C 6 )dialkyl, optionally substituted —C(O)—NH(aryl), optionally substituted —C(O)—N(diaryl), optionally substituted —C(O)—NH(heteroaryl), optionally substituted —C(O)—N(diheteroaryl), optionally substituted
  • R 1 , R 2 and R 5 are each independently selected from H, —OH, —NO 2 , halogen, C 1 -C 6 optionally substituted carboxylic acid group, optionally substituted O—(C 1 -C 6 )alkyl, optionally substituted C 1 -C 6 hydrocarbyl optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted —C(O)—(C 1 -C 6 ) alkyl (ketone), optionally substituted —C(O)—O—(C 1 -C 6 ) alkyl (ester), optionally substituted O—C(O)—(C 1 -C 6 ) alkyl (ester), optionally substituted —C(O)—NH(C 1 -C 6 ) alkyl (urea), optionally substituted —C(O)—N(C 1 -C 6 )dialkyl, optionally substituted —C(O)—NH(aryl),
  • R 3 and R 4 are each independently selected from H, —OH, —NO 2 , halogen, C 1 -C 6 optionally substituted carboxylic acid group, optionally substituted O—(C 1 -C 6 )alkyl, optionally substituted C 1 -C 6 hydrocarbyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted —C(O)—(C 1 -C 6 ) alkyl (ketone), optionally substituted —C(O)—O—(C 1 -C 6 ) alkyl (ester), optionally substituted O—C(O)—(C 1 -C 6 ) alkyl (ester), optionally substituted —C(O)—NH(C 1 -C 6 ) alkyl (urea), optionally substituted —C(O)—N(C 1 -C 6 )dialkyl, optionally substituted —C(O)—NH(aryl), optionally substitute
  • R 6 and R 7 are each independently absent or are selected from H, —OH, —NO 2 , halogen, C 1 -C 6 optionally substituted carboxylic acid group, optionally substituted O—(C 1 -C 6 )alkyl, optionally substituted C 1 -C 6 hydrocarbyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted —C(O)—(C 1 -C 6 ) alkyl (ketone), optionally substituted —C(O)—O—(C 1 -C 6 ) alkyl (ester), optionally substituted O—C(O)—(C 1 -C 6 ) alkyl (ester), optionally substituted —C(O)—NH(C 1 -C 6 ) alkyl (urea), optionally substituted —C(O)—N(C 1 -C 6 )dialkyl, optionally substituted —C(O)—NH(aryl),
  • R 6′ is absent, H, C 1 -C 6 optionally substituted hydrocarbyl group (such as H, CH 3 or CH 2 CH 3 ) or together with R 6 forms ⁇ O;
  • R 7′ is absent, H, optionally substituted hydrocarbyl group (such as H, CH 3 or CH 2 CH 3 ), or together with R forms ⁇ O;
  • R 7′′ is absent, H, —OH, halogen, optionally substituted O—(C 1 -C 6 )alkyl, optionally substituted C 1 -C 6 hydrocarbyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted —C(O)—(C 1 -C 6 ) alkyl (ketone), optionally substituted —C(O)—O—(C 1 -C 6 ) alkyl (ester), optionally substituted O—C(O)—(C 1 -C 6 ) alkyl (ester), optionally substituted —C(O)—NH(C 1 -C 6 ) alkyl (urea), optionally substituted —C(O)—N(C 1 -C 6 )dialkyl, optionally substituted —C(O)—NH(aryl), optionally substituted —C(O)—N(diaryl), optionally substituted —C(O)—
  • R 8′ is absent (when the carbon to which R 8′ is attached and the carbon to which R 6 is attached form an optional double bond), H, CH 3 or CH 2 CH 3 ;
  • R 10 , R 11 , R 12 and R 13 are each independently H, —OH, —NO 2 , halogen, C 1 -C 6 optionally substituted carboxylic acid group, optionally substituted O—(C 1 -C 6 )alkyl, optionally substituted C 1 -C 6 hydrocarbyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted —C(O)—(C 1 -C 6 ) alkyl (ketone), optionally substituted —C(O)—O—(C 1 -C 6 ) alkyl (ester), optionally substituted O—C(O)—(C 1 -C 6 ) alkyl (ester), optionally substituted —C(O)—NH(C 1 -C 6 ) alkyl (urea), optionally substituted —C(O)—N(C 1 -C 6 )dialkyl, optionally substituted —C(O)—NH(
  • R 14 is H, —OH, —NO 2 , halogen, C 1 -C 6 optionally substituted carboxylic acid group, optionally substituted O—(C 1 -C 6 )alkyl, optionally substituted C 1 -C 6 hydrocarbyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted —C(O)—(C 1 -C 6 ) alkyl (ketone), optionally substituted —C(O)—O—(C 1 -C 6 ) alkyl (ester), optionally substituted O—C(O)—(C 1 -C 6 ) alkyl (ester), optionally substituted —C(O)—NH(C 1 -C 6 ) alkyl (urea), optionally substituted —C(O)—N(C 1 -C 6 )dialkyl, optionally substituted —C(O)—NH(aryl), optionally substituted —C(O)
  • Y is optionally substituted (CH 2 ) n group where n is 0, 1 or 2, optionally substituted ⁇ CH— group, a C ⁇ O group, O, S, optionally substituted N—(C 1 -C 6 )alkyl, optionally substituted N-aryl, optionally substituted N-heteroaryl, optionally substituted N-heterocycle, optionally substituted N—C(O)—(C 1 -C 6 )alkyl, optionally substituted N—C(O)-aryl, optionally substituted N—C(O)-heteroaryl, optionally substituted N—C(O)-heterocycle;
  • Z is optionally substituted (CH 2 ) n group where n is 1 or 2, optionally substituted ⁇ CH— group, a C ⁇ O group, O, S, optionally substituted N—(C 1 -C 6 )alkyl, optionally substituted N-aryl, optionally substituted N-heteroaryl, optionally substituted N-heterocycle, optionally substituted N—C(O)—(C 1 -C 6 )alkyl, optionally substituted N—C(O)-aryl, optionally substituted N—C(O)-heteroaryl, optionally substituted N—C(O)-heterocycle.
  • the compound of the invention is at least one GPER agonist recited in PCT Patent Application No. WO 2016/014847, which is incorporated herein in its entirety by reference:
  • Ring A is an aromatic or heteroaromatic five or six membered ring containing one or more heteroatom such as N, O, or S;
  • R 1 is independently selected from SO 2 NH 2 , SO 2 NR a R b , COOH, CONH 2 and CONR a R b , and H.
  • each occurrence of R a and R b is independently selected from H, alkyl (C 1 -C 6 ), alkenyl (C 2 -C 6 ), alkynyl (C 2 -C 6 ), alkoxy (C 2 -C 6 ), cycloalkyl (C 3 -C 7 ), alkylthio, alkylaryl, and aromatic and heteroaromatic rings.
  • the aromatic and heteroaromatic rings can be further substituted with electron withdrawing and donating groups.
  • R a and R b can form a cyclic ring (C 3 -C 7 ) or an aromatic ring optionally containing one or more heteroatoms.
  • aromatic rings can be further substituted with electron withdrawing groups such as halogens, —COOH, —CN, —NO 2 and the like, or electron donating groups such as alkyl groups;
  • R 2 is H, halogen, or a heteroatom such as N, O, or S;
  • Ring A is an aromatic or heteroaromatic ring (5 or 6 membered);
  • Ring B is a six membered saturated or aromatic ring containing N at the indicated position.
  • the nitrogen of ring B optionally can be substituted with an alkyl, aryl, or alkaryl substituent;
  • Ring C is an independently a substituted or unsubstituted carbocyclic ring, bicyclic ring, aromatic ring, fused aromatic rings, or a heteroaromatic ring. Additionally, when it is a carbocyclic ring, it may contain one or more double bonds and one or more heteroatoms such as N, O, or S. It may also have an ⁇ - ⁇ unsaturated ketone function;
  • R 3 is selected independently from H, halogen, —OH, CN, (C 1 -C 6 ) alkyl, (C 2 -C 6 ) alkenyl, (C 2 -C 6 ) alkynyl, (C 1 -C 6 ) alkoxy, (C 3 -C 7 ) cycloalkyl, (C 1 -C 6 ) alkylthio, NR a R b , R a R b or haloalkyl (e.g., CF 3 ).
  • each occurrence of R a and R b is independently hydrogen or (C 1 -C 6 ) alkyl, or R a and R b form a saturated or unsaturated heterocyclic ring containing from 3-7 ring atoms, which ring may optionally contain another heteroatom selected from N, O, and S, and may be optionally substituted by from 1-3 groups which may be the same or different and are selected from (C 1 -C 4 ) alkyl, phenyl, and benzyl; and m is 1-4:
  • Rings D and B is directly connected or connected through a spacer (C 1 -C 2 ). When connected directly, they can be cis or trans with respect to the fusion of Ring B and C.
  • Ring D is an aromatic or heteroaromatic ring containing one or more heteroatoms such as N, O, or S. It can be optionally substituted with R 5 and R 6 groups selected independently, or with —R 6 R 5 or R 5 R 6 ;
  • R 5 is independently selected from H, halogen, electron donating groups, and electron withdrawing groups such as alkyl, haloalkyl, alkoxy, —NO 2 , —SF 5 , —CN, and the like;
  • R c is alkyl, branched alkyl (C 1 -C 10 ), alkoxy, alkylamino, acyl, alkynyl (C 1 -C 8 ) or alkenyl (C 1 -C 6 ).
  • R 1 is selected from the group consisting of carboxyl, carboxamide, carboxyalkyl, carboxyaryl, cyano, nitro, hydroxyl, sulfonyl, sulfonamide, alkylsulfonamide, arylsulfonamide, alkylsulfonyl, aralkylsulfonamide, trifluoromethylsulfonamide, trifluoromethylsulfonyl carboxamide, and sulfonylcarbamide.
  • R 1 is sulfonamide, alkylsulfonamide, or arylsulfonamide.
  • R 2 can be a 3-, 4-, 5-, or 6-membered saturated or aromatic carbon ring or ring system optionally containing one or two heteroatoms selected from N, O, and S, the ring or ring system optionally substituted with one or more substituents selected from the group consisting of cyano, halo, acyl, acyloxy, alkyl, alkoxy, heteroalkyl, alkylester, alkylamido, alkylamino, aryl, aryloxy, arylalkyl, arylester, azido, alkylhalo, alkenyl, alkynyl, alkyl ether, nitro, thiohalo, and thiocyano.
  • substituents selected from the group consisting of cyano, halo, acyl, acyloxy, alkyl, alkoxy, heteroalkyl, alkylester, alkylamido, alkylamino, aryl, aryloxy, arylal
  • R 3 can be H, or a C 1 -C 5 alkyl or cycloalkyl group optionally substituted with one or more of cyano, nitro, and one or more aromatic or heteroaromatic groups containing N, O, or S.
  • a and D can be independently selected from CH, CH 2 , N, and O, and the bond joining them is a single or double bond as appropriate for the selected atoms.
  • X, Y, and Z are independently selected from no atom (i.e., they are absent), CH, C-halogen, N, O, and S;
  • R 2 is a substituent represented by —R d R e Rf or by —R d COR e Rf; wherein R d , R e , and R f are independently selected from a 3-, 4-, 5-, or 6-membered saturated or aromatic carbon ring or ring system optionally containing one or two heteroatoms selected from N, O, and S, the ring or ring system optionally substituted with one or more substituents selected from the group consisting of cyano, halo, acyl, acyloxy, alkyl, alkoxy, heteroalkyl, alkylester, alkylamido, alkylamino, aryl, aryloxy, arylalkyl, arylester, azido, alkylhalo, alkenyl, alkynyl, alkyl ether, nitro, thiohalo, and thiocyano;
  • the compounds of the invention may possess one or more stereocenters, and each stereocenter may exist independently in either the (R) or (S) configuration.
  • compounds described herein are present in optically active or racemic forms.
  • the compounds described herein encompass racemic, optically active, regioisomeric and stereoisomeric forms, or combinations thereof that possess the therapeutically useful properties described herein. Preparation of optically active forms is achieved in any suitable manner, including by way of non-limiting example, by resolution of the racemic form with recrystallization techniques, synthesis from optically active starting materials, chiral synthesis, or chromatographic separation using a chiral stationary phase.
  • a compound illustrated herein by the racemic formula further represents either of the two enantiomers or mixtures thereof, or in the case where two or more chiral center are present, all diastereomers or mixtures thereof.
  • the compounds of the invention exist as tautomers. All tautomers are included within the scope of the compounds recited herein.
  • Compounds described herein also include isotopically labeled compounds wherein one or more atoms is replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature.
  • isotopes suitable for inclusion in the compounds described herein include and are not limited to 2 H, 3 H, 11 C, 13 C, 14 C, 36 Cl, 18 F, 123 I, 125 I, 13 N, 15 N, 15 O, 17 O, 18 O, 32 P, and 35 S. In certain embodiments, substitution with heavier isotopes such as deuterium affords greater chemical stability.
  • Isotopically labeled compounds are prepared by any suitable method or by processes using an appropriate isotopically labeled reagent in place of the non-labeled reagent otherwise employed.
  • the compounds described herein are labeled by other means, including, but not limited to, the use of chromophores or fluorescent moieties, bioluminescent labels, or chemiluminescent labels.
  • the invention further provides pharmaceutical compositions comprising at least one compound of the invention and at least one pharmaceutically acceptable carrier.
  • the pharmaceutical composition is formulated for topical and/or transdermal application. Topical formulations of the compounds contemplated within the invention may be used for skin darkening, as well as for treating skin conditions or diseases associated with hypopigmentation. These conditions or diseases include, but are not limited to, acne, eczema, chemical, sun and thermal burn scars, and lupus.
  • the invention provides topical formulations comprising at least one sun-blocking agent and at least one compound of the invention.
  • the topical formulation comprises a formulated sunblock or sunscreen lotion and at least one compound of the invention.
  • the at least one sun-blocking agent allows for protection against UV light damage caused by the natural sun light, and the one or more compounds of the invention allow for darkening of the skin.
  • the melanin production triggered by the at least one compound of the invention further protects the skin against UV light damage caused by the natural sun light.
  • the topical formulation protects against UV-induced skin damage and/or aging.
  • kits useful within any of the methods of the invention described herein comprise components useful in any of the methods described herein, including for example, compositions and methods for modulating skin pigmentation in a mammalian subject, such as a human, one or more containers (e.g., test tube, cell culture dish, cell culture plate, cell culture flask, cell culture bag) for containing a component of any of the embodiments of the invention described elsewhere herein, and instructional materials.
  • a mammalian subject such as a human
  • containers e.g., test tube, cell culture dish, cell culture plate, cell culture flask, cell culture bag
  • salts embraces addition salts of free acids or bases that are useful within the methods of the invention.
  • pharmaceutically acceptable salt refers to salts that possess toxicity profiles within a range that affords utility in pharmaceutical applications.
  • the salts are pharmaceutically acceptable salts.
  • Pharmaceutically unacceptable salts may nonetheless possess properties such as high crystallinity, which have utility in the practice of the present invention, such as for example utility in process of synthesis, purification or formulation of compounds useful within the methods of the invention.
  • Suitable pharmaceutically acceptable acid addition salts may be prepared from an inorganic acid or from an organic acid.
  • inorganic acids include sulfate, hydrogen sulfate, hydrochloric, hydrobromic, hydriodic, nitric, carbonic, sulfuric, and phosphoric acids (including hydrogen phosphate and dihydrogen phosphate).
  • organic acids may be selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which include formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, 4-hydroxybenzoic, phenylacetic, mandelic, embonic (or pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic, sulfanilic, 2-hydroxyethanesulfonic, trifluoromethanesulfonic, p-toluenesulfonic, cyclohexylaminosulfonic, stearic, alginic, (3-hydroxybutyric, salicy
  • Suitable pharmaceutically acceptable base addition salts of compounds of the invention include, for example, ammonium salts and metallic salts including alkali metal, alkaline earth metal and transition metal salts such as, for example, calcium, magnesium, potassium, sodium and zinc salts.
  • Pharmaceutically acceptable base addition salts also include organic salts made from basic amines such as, for example, N,N′-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (or N-methylglucamine) and procaine. All of these salts may be prepared from the corresponding compound by reacting, for example, the appropriate acid or base with the compound.
  • the invention includes a method of modulating skin pigmentation in a mammalian subject, such as a human.
  • the method of the invention treats or prevents skin hypopigmentation in the subject.
  • the compound is administered topically or transdermally to the subject.
  • the method comprises administering to the subject a therapeutically effective amount of a compound that binds to a non-classical hormone receptor present on a melanocyte and does not bind to a canonical sex steroid hormone receptor, such as the estrogen receptor.
  • the non-classical hormone receptor comprises GPER.
  • the non-classical hormone receptor comprises GPER1.
  • the compound is a GPER agonist and increases, or prevents further loss of, skin pigmentation.
  • the subject is suffering from a condition comprising at least one selected from the group consisting of pigmentary changes associated with oral contraceptive use, pregnancy, and endogenous estrogens in females (such as melasma and chloasma); solar lentigo; acne; eczema; chemical, sun, and thermal burn scars; and lupus.
  • a condition comprising at least one selected from the group consisting of pigmentary changes associated with oral contraceptive use, pregnancy, and endogenous estrogens in females (such as melasma and chloasma); solar lentigo; acne; eczema; chemical, sun, and thermal burn scars; and lupus.
  • compositions of the invention will vary, depending upon the identity, size, and condition of the subject treated.
  • the composition may comprise between about 0.005% and about 100% (w/w) of the active agent, or any fractions or multiples thereof.
  • compositions are principally directed to pharmaceutical compositions that are suitable for ethical administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and perform such modification with merely ordinary, if any, experimentation. Subjects to which administration of the pharmaceutical compositions of the invention is contemplated include, but are not limited to, humans and other primates, mammals including commercially relevant mammals such as cattle, pigs, horses, sheep, cats, and dogs.
  • compositions comprising a compound contemplated within the invention are formulated using one or more pharmaceutically acceptable excipients or carriers.
  • the pharmaceutical compositions of the invention comprise a therapeutically effective amount of at least one compound contemplated within the invention and a pharmaceutically acceptable carrier.
  • Pharmaceutically acceptable carriers include, but are not limited to, glycerol, water, saline, ethanol and other pharmaceutically acceptable salt solutions such as phosphates and salts of organic acids. Examples of these and other pharmaceutically acceptable carriers are described in Remington's Pharmaceutical Sciences (1991, Mack Publication Co., New Jersey).
  • the carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils.
  • the proper fluidity may be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • Prevention of the action of microorganisms may be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like.
  • isotonic agents for example, sugars, sodium chloride, or polyalcohols such as mannitol and sorbitol, are included in the composition.
  • Prolonged absorption of the injectable compositions may be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate or gelatin.
  • Formulations may be employed in admixtures with conventional excipients, i.e., pharmaceutically acceptable organic or inorganic carrier substances suitable for oral, parenteral, nasal, intravenous, subcutaneous, enteral, or any other suitable mode of administration, known to the art.
  • the administration comprises topical administration.
  • the pharmaceutical preparations may be sterilized and if desired mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure buffers, coloring, flavoring and/or aromatic substances and the like. They may also be combined where desired with other active agents, e.g., other analgesic agents.
  • additional ingredients include, but are not limited to, one or more of the following: excipients; surface active agents; dispersing agents; inert diluents; granulating and disintegrating agents; binding agents; lubricating agents; sweetening agents; flavoring agents; coloring agents; preservatives; physiologically degradable compositions such as gelatin; aqueous vehicles and solvents; oily vehicles and solvents; suspending agents; dispersing or wetting agents; emulsifying agents, demulcents; buffers; salts; thickening agents; fillers; emulsifying agents; antioxidants; antibiotics; antifungal agents; stabilizing agents; and pharmaceutically acceptable polymeric or hydrophobic materials.
  • compositions of the invention are known in the art and described, for example in Genaro, ed. (1985, Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.), which is incorporated herein by reference.
  • the composition of the invention may comprise a preservative from about 0.005% to 2.0% by total weight of the composition.
  • the preservative is used to prevent spoilage in the case of exposure to contaminants in the environment.
  • Examples of preservatives useful in accordance with the invention included but are not limited to those selected from the group consisting of benzyl alcohol, sorbic acid, parabens, imidurea and combinations thereof.
  • An illustrative preservative is a combination of about 0.5% to 2.0% benzyl alcohol and 0.05% to 0.5% sorbic acid.
  • the composition may include an antioxidant and a chelating agent that inhibit the degradation of the compound.
  • Selected antioxidants for some compounds are BHT, BHA, ⁇ -tocopherol and ascorbic acid in the exemplary range of about 0.01% to 0.3%, such as BHT in the range of 0.03% to 0.1% by weight by total weight of the composition.
  • the chelating agent is present in an amount of from 0.01% to 0.5% by weight by total weight of the composition.
  • Illustrative chelating agents include edetate salts (e.g. disodium edetate) and citric acid in the weight range of about 0.01% to 0.20%, or in the range of 0.02% to 0.10% by weight by total weight of the composition.
  • the chelating agent is useful for chelating metal ions in the composition which may be detrimental to the shelf life of the formulation. While BHT and disodium edetate are illustrative antioxidant and chelating agent respectively for some compounds, other suitable and equivalent antioxidants and chelating agents may be substituted as would be known to those skilled in the art.
  • composition comprising a compound contemplated within the invention can be administered to a mammal as frequently as several times daily, or it may be administered less frequently, such as once a day, once a week, once every two weeks, once a month, or even less frequently, such as once every several months or even once a year or less.
  • the amount of compound dosed per day may be administered, in non-limiting examples, every day, every other day, every 2 days, every 3 days, every 4 days, or every 5 days.
  • a 0.5-5 mg per day dose may be initiated on Monday with a first subsequent 0.5-5 mg per day dose administered on Wednesday, a second subsequent 0.5-5 mg per day dose administered on Friday, and so on.
  • the frequency of the dose will be readily apparent to the skilled artisan and will depend upon any number of factors, such as, but not limited to, the type and severity of the disease being treated, the type and age of the animal, and so forth.
  • stratum corneum layer of the epidermis An obstacle for topical administration of pharmaceuticals is the stratum corneum layer of the epidermis.
  • the stratum corneum is a highly resistant layer comprised of protein, cholesterol, sphingolipids, free fatty acids and various other lipids, and includes cornified and living cells.
  • One of the factors that limit the penetration rate (flux) of a compound through the stratum corneum is the amount of the active substance that can be loaded or applied onto the skin surface. The greater the amount of active substance applied per unit of area of the skin, the greater the concentration gradient between the skin surface and the lower layers of the skin, and in turn the greater the diffusion force of the active substance through the skin. Therefore, a formulation containing a greater concentration of the active substance is more likely to result in penetration of the active substance through the skin, and more of it, and at a more consistent rate, than a formulation having a lesser concentration, all other things being equal.
  • Formulations suitable for topical administration include, but are not limited to, liquid or semi-liquid preparations such as liniments, lotions, oil-in-water or water-in-oil emulsions such as creams, ointments or pastes, and solutions or suspensions.
  • Such formulations may be applied to the skin directly or through the use of swabs, applicators, spatulas and the like, as well as in the form of transdermal patches.
  • the patch minimizes loss of pharmaceuticals through washing, friction, scratching and/or rubbing of the skin.
  • the patch increases absorption of the pharmaceutical through the skin, while minimizing the exposure of the skin to the pharmaceutical.
  • Topically administrable formulations may, for example, comprise from about 0.005% to about 10% (w/w) active ingredient, although the concentration of the active ingredient may be as high as the solubility limit of the active ingredient in the solvent.
  • Formulations for topical administration may further comprise one or more of the additional ingredients described herein.
  • Enhancers of permeation may be used. These materials increase the rate of penetration of drugs across the skin. Typical enhancers in the art include ethanol, glycerol monolaurate, PGML (polyethylene glycol monolaurate), dimethylsulfoxide, and the like. Other enhancers include oleic acid, oleyl alcohol, ethoxydiglycol, laurocapram, alkanecarboxylic acids, dimethylsulfoxide, polar lipids, or N-methyl-2-pyrrolidone.
  • compositions of the invention may contain liposomes.
  • the composition of the liposomes and their use are known in the art (for example, U.S. Pat. No. 6,323,219).
  • the pharmaceutical composition of the invention may be optionally combined with other ingredients such as adjuvants, anti-oxidants, chelating agents, surfactants, foaming agents, wetting agents, emulsifying agents, viscosifiers, buffering agents, preservatives, and the like.
  • a permeation or penetration enhancer is included in the composition and is effective in improving the percutaneous penetration of the active ingredient into and through the stratum corneum with respect to a composition lacking the permeation enhancer.
  • compositions may further comprise a hydrotropic agent, which functions to increase disorder in the structure of the stratum corneum, and thus allows increased transport across the stratum corneum.
  • hydrotropic agents such as isopropyl alcohol, propylene glycol, or sodium xylene sulfonate, are known to those of skill in the art.
  • the topically active pharmaceutical composition of the invention should be applied in an amount effective to affect desired changes.
  • amount effective shall mean an amount sufficient to cover the region of skin surface where a change is desired.
  • An active compound should be present in the amount of from about 0.0001% to about 15% by weight volume of the composition; or in an amount from about 0.0005% to about 5% of the composition; or in an amount of from about 0.005% to about 1% of the composition.
  • Such compounds may be synthetically-or naturally derived.
  • Controlled- or sustained-release formulations of a pharmaceutical composition of the invention may be made using conventional technology.
  • the dosage forms to be used can be provided as slow or controlled-release of one or more active ingredients therein using, for example, hydropropylmethyl cellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, or microspheres or a combination thereof to provide the desired release profile in varying proportions.
  • Suitable controlled-release formulations known to those of ordinary skill in the art, including those described herein, can be readily selected for use with the pharmaceutical compositions of the invention.
  • single unit dosage forms suitable for topical administration such as liniments, lotions, oil-in-water or water-in-oil emulsions such as creams, ointments or pastes, transdermal patches, and solutions or suspensions that are adapted for controlled-release are encompassed by the present invention.
  • controlled-release pharmaceutical products have a common goal of improving drug therapy over that achieved by their non-controlled counterparts.
  • the use of an optimally designed controlled-release preparation in medical treatment is characterized by a minimum of drug substance being employed to cure or control the condition in a minimum amount of time.
  • Advantages of controlled-release formulations include extended activity of the drug, reduced dosage frequency, and increased patient compliance.
  • controlled-release formulations can be used to affect the time of onset of action or other characteristics, such as blood level of the drug, and thus can affect the occurrence of side effects.
  • controlled-release formulations are designed to initially release an amount of drug that promptly produces the desired therapeutic effect, and gradually and continually release of other amounts of drug to maintain this level of therapeutic effect over an extended period of time.
  • the drug In order to maintain this constant level of drug in the body, the drug must be released from the dosage form at a rate that will replace the amount of drug being metabolized and excreted from the body.
  • Controlled-release of an active ingredient can be stimulated by various inducers, for example pH, temperature, enzymes, water, or other physiological conditions or compounds.
  • controlled-release component in the context of the present invention is defined herein as a compound or compounds, including, but not limited to, polymers, polymer matrices, gels, permeable membranes, liposomes, or microspheres or a combination thereof that facilitates the controlled-release of the active ingredient.
  • the formulations of the present invention may be, but are not limited to, short-term, rapid-offset, as well as controlled, for example, sustained release, delayed release and pulsatile release formulations.
  • sustained release is used in its conventional sense to refer to a drug formulation that provides for gradual release of a drug over an extended period of time, and that may, although not necessarily, result in substantially constant blood levels of a drug over an extended time period.
  • the period of time may be as long as a month or more and should be a release that is longer that the same amount of agent administered in bolus form.
  • the compounds may be formulated with a suitable polymer or hydrophobic material which provides sustained release properties to the compounds.
  • the compounds for use the method of the invention may be administered in the form of microparticles, for example, by injection or in the form of wafers or discs by implantation.
  • the compounds of the invention are administered to a patient, alone or in combination with another pharmaceutical agent, using a sustained release formulation.
  • delayed release is used herein in its conventional sense to refer to a drug formulation that provides for an initial release of the drug after some delay following drug administration and that may, although not necessarily, includes a delay of from about 10 minutes up to about 12 hours.
  • pulsatile release is used herein in its conventional sense to refer to a drug formulation that provides release of the drug in such a way as to produce pulsed plasma profiles of the drug after drug administration.
  • immediate release is used in its conventional sense to refer to a drug formulation that provides for release of the drug immediately after drug administration.
  • short-term refers to any period of time up to and including about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 40 minutes, about 20 minutes, or about 10 minutes and any or all whole or partial increments thereof after drug administration after drug administration.
  • rapid-offset refers to any period of time up to and including about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 40 minutes, about 20 minutes, or about 10 minutes, and any and all whole or partial increments thereof after drug administration.
  • reaction conditions including but not limited to reaction times, reaction size/volume, and experimental reagents, such as solvents, catalysts, pressures, atmospheric conditions, e.g., nitrogen atmosphere, and reducing/oxidizing agents, with art-recognized alternatives and using no more than routine experimentation, are within the scope of the present application.
  • cAMP ELISA was performed on primary human melanocytes using the Cyclic AMP XP Assay Kit (Cell Signaling Technology, #4339) following manufacturer instructions.
  • Adherent cells were treated with 1 ⁇ M doses of E2 overnight, washed once with DPBS, and lysed with 1% NP-40 buffer (150 mM NaCl, 50 mM Tris, pH 7.5, 1 mM EDTA, and 1% NP-40) containing 1 ⁇ protease inhibitors (Roche) and 1 ⁇ phosphatase inhibitors (Roche). Lysates were quantified (Bradford assay), normalized, reduced, denatured (95° C.) and resolved by SDS gel electrophoresis on 4-15% Tris/Glycine gels (Bio-Rad).
  • Resolved protein was transferred to PVDF membranes (Millipore) using a Semi-Dry Transfer Cell (Bio-Rad), blocked in 5% dry milk in TBS-T and probed with primary antibodies recognizing MITF (Cell Signaling Technology. #12590, 1:1000), pCREB (Cell Signaling Technology, #9198, 1:1000), CREB (Cell Signaling Technology, #9104, 1:1000), and ⁇ -Actin (Cell Signaling Technology, #3700, 1:4000). After incubation with the appropriate secondary antibody, proteins were detected using either Luminata Crescendo Western HRP Substrate (Millipore) or ECL Western Blotting Analysis System (GE Healthcare).
  • Formalin-fixed paraffin embedded tissue was sectioned at 5 uM and collected on superfrost plus slides (Fisher), and subjected to Fontana-Masson stain for melanin. Briefly, sections were deparaffinized, rehydrated, and incubated in the following solutions: 2.5% aqueous silver nitrate for 10 min, 0.1% aqueous gold chloride for 15 min, and 5% aqueous sodium thiosulfate for 5 min. Distilled deionized water was used for rinsing and incubations were done at room temperature except for silver nitrate at 60° C. Slides were counterstained with 0.1% nuclear fast red Kemechtrot for 5 min, dehydrated, cleared, and coverslipped using MM24 mounting media (Leica). All staining reagents were from Polyscientific R & D Corporation.
  • FFPE Formalin fixed paraffin embedded
  • Tissue sections from organotypic culture were stained using methods described elsewhere herein. In terms of quantitation, briefly, 20 ⁇ photomicrograph images of representative tissue sections were taken using the Zeiss Axiophot microscope. Tiff files of the images were saved and transferred to Adobe PHOTOSHOP® where pixels corresponding to Fontana-Masson staining and epidermal counter stain were selected using the color selection tool. Images corresponding to the single specific color were then analyzed using FIJI (Image J) to determine the number of pixels in each sample. The numbers of pixels representing Fontana-Masson staining were normalized to the total amount epidermal counter staining. Final ratios Fontana-Masson staining in the epidermis were set relative to amount of staining in vehicle treated controls.
  • FIJI Image J
  • mRNA was extracted from melanocytes according to the RNeasy Mini Kit protocol (Qiagen), and reverse transcribed to cDNA using the High Capacity RNA-to cDNA kit (Applied Biosystems). Quantitative PCR of the resulting cDNA was carried out using Power SYBR Green Master Mix (Applied Biosystems) and gene-specific primers, in triplicate, on a ViiA 7 Real-Time PCR System (Life Technologies).
  • B-Actin forward 5′-CAT GTA CGT TGC TAT CCA GGC-3′ (SEQ ID NO: 1).
  • B-Actin reverse 5′-CTC CTT AAT GTC ACG CAC GAT-3′ (SEQ ID NO:2);
  • ER-A forward 5′-AAA GGT GGG ATA CGA AAA GAC C-3′ (SEQ ID NO:3);
  • ER-A reverse 5′-AGC ATC CAA CAA GGC ACT GA-3′ (SEQ ID NO:4);
  • ER-B forward 5′-GGC TGC GAG AAA TAA CTG CC-3′ (SEQ ID NO:5);
  • ER-B reverse 5′-AAT GCG GAC ACG TGC TTT TC-3′ (SEQ ID NO:6);
  • PGR forward 5′-AGG TCT ACC CGC CCT ATC TC-3′ (SEQ ID NO:7);
  • PGR reverse 5′-AGT AGT TGT GCT GCC CTT
  • shRNAs were expressed from the GIPZ vector (Open Biosystems): shPAR7.3 (V3LHS_364596, 5′-TGT GGT AGA GAA GAG CTG G-3′; SEQ ID NO: 13), shGPER.1 (V2LHS_132008, 5′-TCC TTC TCC TCT TTA ACT C-3′; SEQ ID NO: 14), shGPER.3 (V3LHS_390319, 5′-TGA TGA AGT ACA GGT CGG G-3′; SEQ ID NO: 15).
  • Guide RNAs were designed using software tools provided on the website www dot genome-engineering dot org/.
  • Guide RNAs were subsequently cloned into lentiCRISPRv2 (Addgene #52961) according to the accompanying protocol.
  • Guide RNA sequences are as follows: lentiCRISPR GFP, 5′-GAA GTT CGA GGG CGA CAC CC-3′ (SEQ ID NO: 16); lentiCRISPR GPER.1, 5′-ACAGGCCGATCACGTACTGC-3′ (SEQ ID NO: 17); lentiCRISPR GPER.2, 5′-GAG CAC CAG CAG TAC GTG AT-3′ (SEQ ID NO: 18).
  • * denotes a P-value of less than 0.05 in an unpaired, two-tailed Students T-Test.
  • Organotypic skin grafts containing MCs were established using modifications to previously detailed methods (Ridky, et al., Nature Med. 2010, 16, 1450-1455; Chudnovsky, et al., Nature Gen. 2005, 37, 745-749).
  • KGM Keratinocyte Growth Media
  • MXSM Melanocyte Xenograft Seeding Media
  • MXSM is a 1:1 mixture of KGM, lacking cholera toxin, and Keratinocyte Media 50/50 (Gibco) containing 2% FBS, 1.2 mM calcium chloride, 100 nM Et-3 (endothelin 3), 10 ng/mL rhSCF (recombinant human stem cell factor), and 4.5 ng/mL r-basic FGF (recombinant basic fibroblast growth factor).
  • 1.5 ⁇ 10 5 melanocytes and 5.0 ⁇ 10 5 keratinocytes were suspended in 80 ⁇ L MXSM, seeded onto the dermis, and incubated at 37° C. for 8 days at the air-liquid interface.
  • melanocytes isolated from body sites other than foreskin also responded similarly to estrogen, melanocytes from adult facial skin were treated with this sex hormone; responses observed were similar to those observed with the foreskin melanocytes ( FIG. 2F ).
  • Example 3 Primary Human Melanocytes do not Express Nuclear Estrogen Receptor (ER), and Respond to Estrogen Via Altered cAMP Signaling
  • estrogen, ] and ⁇ -MSH converge on the canonical melanin production pathway at the level of adenylate cyclase to modulate melanin synthesis.
  • pregnancy-associated hyperpigmentation is not observed all over the body, but is characteristically limited to specific areas where melanocyte density or UV radiation exposure is highest, including the face, genital, and areolar regions.
  • GPCR G protein-coupled receptor
  • Tamoxifen an ER antagonist
  • GPER GABA-like steroid receptor
  • Tamoxifen an ER antagonist
  • tamoxifen blocks estrogen receptor function and thus might have been predicted to limit melanin synthesis.
  • tamoxifen acts as a GPER agonist. Tamoxifen treatment resulted in a dose-dependent increase in melanin production to levels comparable to those observed with native estrogen ( FIG. 7A ), consistent with the model in which GPER activation promotes melanin synthesis.
  • Most oral contraceptives incorporate an estrogen derivative (ethinyl estradiol) for the estrogen component.
  • ethinyl estradiol an estrogen derivative for the estrogen component.
  • Treatment of primary human MCs with ethinyl estradiol also resulted in a dose dependent increase in melanin production, indistinguishable from native estrogen at an equal concentration ( FIG. 7B ).
  • Example 7 Compounds of the Invention Induce pCREB in Human Primary Melanocytes
  • Solvents used for extraction and purification were HPLC grade from Fisher Scientific. Unless otherwise indicated, all reactions were run under an inert atmosphere of argon. Anhydrous tetrahydrofuran, dichloromethane and toluene were obtained via passage through an activated alumina column. Commercial reagents were used as received.
  • VWR pre-coated silica gel plates 250 ⁇ m, 60 F254 were used for analytical TLC. Spots were visualized using 254 nm ultraviolet light with either potassium permanganate or vanillin stains as visualizing agents. Chromatographic purifications were performed on Sorbent Technologies silica gel (particle size 32-63 microns).
  • Infrared spectra were recorded on a NaCl plate using a Perkin-Elmer 1600 series Fourier transform spectrometer.
  • G-1 (20 mg, 0.0486 mmol), sodium tertbutoxide (6.5 mg, 0.0680 mmol), JohnPhos (1.5 mg, 4.86 ⁇ mol) and tris(dibenzylideneacetone)dipalladium (2.2 mg, 2.43 ⁇ mol) were added to a vial which was evacuated and back-filled with argon 3 times. Toluene (0.5 mL) and benzylamine (6.0 ⁇ L, 0.0584 mmol) were then added and the mixture was heated to 110° C. for 24 h.
  • CMPD12 10 mg, 0.0300 mmol, 62%) as a yellow oil and a racemic mixture of enantiomers.
  • hydroxylamine hydrochloride 13 mg, 0.190 mmol was added to a solution of G-1 (60 mg, 0.146 mmol) and pyridine (15 mg, 0.190 mmol) in dioxane (1.5 mL) and the solution was heated to 80° C. for 16 h. The solution was cooled to rt and purification through preparative TLC (2% MeOH in CH 2 Cl 2 ) gave oxime CMPD6 (18 mg, 0.0422 mmol, 29%) as an off-white amorphous solid and a racemic mixture of enantiomers.
  • CMPD10 24 mg, 0.0530 mmol
  • 2,2-dimethoxypropane (0.12 mL, 0.971 mmol)
  • acetone 0.7 mL
  • p-Toluenesulfonic acid monohydrate 3.0 mg, 0.0162 mmol
  • CMPD10 24 mg, 0.0530 mmol
  • 2,2-dimethoxypropane (0.12 mL, 0.971 mmol)
  • acetone 0.7 mL
  • acetone 0.7 mL
  • the solution was concentrated in vacuo, redissolved in EtOAc (10 mL) and washed with sat. aqueous NaHCO 3 (10 mL).
  • the aqueous phase was extracted with EtOAc (2 ⁇ 10 mL) and the combined organic phases were dried (Na 2 SO 4 ).
  • Chromatography (1:2 hexanes/Et 2 O) gave ketal CMPD9 (20 mg, 0.0415 mmol,
  • Tissue generated elsewhere herein is orthotopically xenografted onto immunocompromised mice, allowing analysis of human skin in vivo. Hormones are administered subcutaneously or topically to demonstrate the necessity of GPER to mediate hormone-induced changes in pigmentation. The effects of the specific agonists are evaluated.
  • melanocytes are isolated from various body sites and donors of different age, sex, and race. 3-D organotypic cultures are established with melanocytes from these sources, and the necessity and sufficiency of GPER signaling is evaluated.
  • the G ⁇ subunit activated by the receptor is identified.
  • agonist-promoted [ 35 S]GTP ⁇ S exchange assays are used, followed by immuno-enrichment of specific G protein subgroups.
  • G protein-coupled receptor signaling drives guanine nucleotide exchange on the G ⁇ subunit, switching GDP for GTP.
  • [ 35 S]GTP ⁇ S is an isotopic, non-hydrolysable analogue of GTP.
  • Membrane preparations from primary human melanocytes are isolated and stimulated with GPER agonist, as well as vehicle controls, in the presence of [ 35 S]GTP ⁇ S.
  • Gs and Gi subunits are immunoprecipitated from the samples, and [ 35 S]GTP ⁇ S binding is measured using a scintillation spectrophotometer. Melanocytes with ablated GPER are used as negative controls in these assays.
  • adenylate cyclase activity which functions to produce cAMP.
  • adenylate cyclase activity is analyzed in the presence of estrogen using cAMP ELISA, relative to vehicle-treated controls.
  • Western analysis is used to examine downstream changes in pCREB and MITF relative to vehicle-treated controls.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Dermatology (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Epidemiology (AREA)
  • Biochemistry (AREA)
  • Toxicology (AREA)
  • Birds (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Plural Heterocyclic Compounds (AREA)
  • Cosmetics (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
US15/562,776 2015-04-06 2016-04-06 Compositions and methods for increasing, or preventing or reversing loss of, skin pigmentation in a mammalian subject Abandoned US20180273482A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US15/562,776 US20180273482A1 (en) 2015-04-06 2016-04-06 Compositions and methods for increasing, or preventing or reversing loss of, skin pigmentation in a mammalian subject

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201562143597P 2015-04-06 2015-04-06
US201562216792P 2015-09-10 2015-09-10
PCT/US2016/026199 WO2016164450A1 (en) 2015-04-06 2016-04-06 Compositions and methods for increasing, or preventing or reversing loss of, skin pigmentation in a mammalian subject
US15/562,776 US20180273482A1 (en) 2015-04-06 2016-04-06 Compositions and methods for increasing, or preventing or reversing loss of, skin pigmentation in a mammalian subject

Publications (1)

Publication Number Publication Date
US20180273482A1 true US20180273482A1 (en) 2018-09-27

Family

ID=57072935

Family Applications (3)

Application Number Title Priority Date Filing Date
US15/562,776 Abandoned US20180273482A1 (en) 2015-04-06 2016-04-06 Compositions and methods for increasing, or preventing or reversing loss of, skin pigmentation in a mammalian subject
US15/562,725 Active US11236074B2 (en) 2015-04-06 2016-04-06 Compositions and methods for decreasing, or preventing or reversing gain of, skin pigmentation in a mammalian subject
US17/480,948 Active US11987576B2 (en) 2015-04-06 2021-09-21 Compositions and methods for decreasing, or preventing or reversing gain of, skin pigmentation in a mammalian subject

Family Applications After (2)

Application Number Title Priority Date Filing Date
US15/562,725 Active US11236074B2 (en) 2015-04-06 2016-04-06 Compositions and methods for decreasing, or preventing or reversing gain of, skin pigmentation in a mammalian subject
US17/480,948 Active US11987576B2 (en) 2015-04-06 2021-09-21 Compositions and methods for decreasing, or preventing or reversing gain of, skin pigmentation in a mammalian subject

Country Status (8)

Country Link
US (3) US20180273482A1 (de)
EP (2) EP3280410A4 (de)
JP (1) JP2018517665A (de)
KR (1) KR20170134636A (de)
CN (1) CN107613971A (de)
AU (2) AU2016246688B2 (de)
CA (1) CA2981785A1 (de)
WO (2) WO2016164460A2 (de)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10934277B2 (en) 2018-07-21 2021-03-02 Linnaeus Therapeutics, Inc. Enantiomerically purified GPER agonist for use in treating disease states and conditions

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12037327B2 (en) 2018-05-04 2024-07-16 Saint Louis University Compounds and methods targeting GPER for treatment of diseases associated with calcium
US11883523B2 (en) 2018-09-18 2024-01-30 Mary Kay Inc. Plant extracts to reduce the appearance of hyperpigmented skin or to lighten skin

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5227459A (en) * 1990-05-18 1993-07-13 Yale University Synthetic melanin
WO2004072046A2 (en) * 2003-02-12 2004-08-26 Carex S.A. Quinoline derivatives and their use for modulation of lxr activity
US20080167334A1 (en) * 2005-08-04 2008-07-10 Prossnitz Eric R Compounds for binding to ERalpha/beta and GPR30, methods of treating disease states and conditions mediated through these receptors and identification thereof
US20090246156A1 (en) * 2008-03-31 2009-10-01 Audrey Kunin Anti-acne sunscreen composition
US20100183528A1 (en) * 2008-12-17 2010-07-22 Harmony Laboratories, Inc. Acne treatment powder foundation

Family Cites Families (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB1051693A (de) * 1962-07-09
US3275622A (en) * 1963-12-18 1966-09-27 Syntex Corp 19-alkylidene-delta4-3, 20-keto pregnanes
US6323219B1 (en) 1998-04-02 2001-11-27 Ortho-Mcneil Pharmaceutical, Inc. Methods for treating immunomediated inflammatory disorders
US8426457B2 (en) 2003-03-13 2013-04-23 Medicis Pharmaceutical Corporation Methods of improving skin quality
CA2578529A1 (en) 2004-08-18 2006-02-23 Ace Aps Cosmetic and pharmaceutical compositions comprising ace inhibitors and/or angiotensin ii receptor antagonists
WO2008121850A2 (en) 2007-03-30 2008-10-09 University Of Rochester Small-molecule modulators of melanin expression
KR101199672B1 (ko) 2008-02-01 2012-11-09 마리 케이 인코포레이티드 방향성 피부활성 성분을 사용하여 피부를 치료하는 방법
BRPI1008752B1 (pt) 2009-02-16 2020-02-11 Nogra Pharma Limited Compostos alquilamido, composições farmacêuticas e usos dos mesmos
WO2012122101A1 (en) 2011-03-04 2012-09-13 Fox Chase Cancer Center Methods for inhibiting proliferation and inducing apoptosis of cancer cells expressing the g protein-coupled estrogen receptor gpr30
CN104083368B (zh) 2014-05-19 2016-08-17 中山大学 G-1在制备基于g蛋白偶联受体30的三阴性乳腺癌靶向药物方面的应用
WO2016014847A1 (en) 2014-07-23 2016-01-28 Northeastern University Ligands for alpha-7 nicotinic acetylcholine receptors and methods of treating neurological and inflammatory conditions

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5227459A (en) * 1990-05-18 1993-07-13 Yale University Synthetic melanin
WO2004072046A2 (en) * 2003-02-12 2004-08-26 Carex S.A. Quinoline derivatives and their use for modulation of lxr activity
US20080167334A1 (en) * 2005-08-04 2008-07-10 Prossnitz Eric R Compounds for binding to ERalpha/beta and GPR30, methods of treating disease states and conditions mediated through these receptors and identification thereof
US20090246156A1 (en) * 2008-03-31 2009-10-01 Audrey Kunin Anti-acne sunscreen composition
US20100183528A1 (en) * 2008-12-17 2010-07-22 Harmony Laboratories, Inc. Acne treatment powder foundation

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10934277B2 (en) 2018-07-21 2021-03-02 Linnaeus Therapeutics, Inc. Enantiomerically purified GPER agonist for use in treating disease states and conditions
US11760749B2 (en) 2018-07-21 2023-09-19 Linnaeus Therapeutics, Inc. Enantiomerically purified GPER agonist for use in treating disease states and conditions

Also Published As

Publication number Publication date
EP3280410A4 (de) 2018-12-19
EP3280702A4 (de) 2018-12-12
US11987576B2 (en) 2024-05-21
US20180354935A1 (en) 2018-12-13
KR20170134636A (ko) 2017-12-06
AU2016246688A1 (en) 2017-10-26
CA2981785A1 (en) 2016-10-13
CN107613971A (zh) 2018-01-19
EP3280410A2 (de) 2018-02-14
US11236074B2 (en) 2022-02-01
WO2016164460A2 (en) 2016-10-13
JP2018517665A (ja) 2018-07-05
WO2016164460A3 (en) 2016-11-24
US20220017497A1 (en) 2022-01-20
WO2016164450A1 (en) 2016-10-13
EP3280702A1 (de) 2018-02-14
AU2016246688B2 (en) 2020-11-12
AU2016246681A1 (en) 2017-10-26

Similar Documents

Publication Publication Date Title
US11987576B2 (en) Compositions and methods for decreasing, or preventing or reversing gain of, skin pigmentation in a mammalian subject
EP3458454B1 (de) 2-hydroxy-n-(benzo[d]thiazol-2-yl)benzamid-derivate als mitochondrien-entkoppler zur behandlung von stoffwechselerkrankungen und krebs
US11369618B2 (en) Compounds, compositions and methods for prevention and/or treatment of cancer
US12102633B2 (en) Spirolactone compounds
JP2014529628A (ja) 皮膚色素沈着を調節するための方法
US9834525B2 (en) Inhibitors of paxillin function and related compositions and methods
WO2021162463A9 (ko) 플라본-레스베라트롤 접합체 화합물을 포함하는 아토피 피부염 예방, 치료 및 개선용 조성물
AU2015346022B2 (en) Methods and compositions for enhancing skin pigmentation
Wei et al. Discovery of cinnamamide-barbiturate hybrids as a novel class of Nrf2 activator against myocardial ischemia/reperfusion injury
US9078852B2 (en) Retinaldehyde in the treatment of obesity, diabetes and other conditions
KR101712184B1 (ko) Nrf2 활성화 효능의 세스퀴터핀 화합물 및 이를 유효성분으로 포함하는 약학조성물
WO2019118855A1 (en) Compounds and compositions for prevention and/or treatment of cancer

Legal Events

Date Code Title Description
AS Assignment

Owner name: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF

Free format text: CONFIRMATORY LICENSE;ASSIGNOR:UNIVERSITY OF PENNSYLVANIA;REEL/FRAME:045992/0745

Effective date: 20180409

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: FINAL REJECTION MAILED

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION