WO2022204305A2 - Peptides and methods for reducing skin pigmentation - Google Patents

Peptides and methods for reducing skin pigmentation Download PDF

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Publication number
WO2022204305A2
WO2022204305A2 PCT/US2022/021579 US2022021579W WO2022204305A2 WO 2022204305 A2 WO2022204305 A2 WO 2022204305A2 US 2022021579 W US2022021579 W US 2022021579W WO 2022204305 A2 WO2022204305 A2 WO 2022204305A2
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WIPO (PCT)
Prior art keywords
derivative
gly
val
ala
leu
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PCT/US2022/021579
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English (en)
French (fr)
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WO2022204305A3 (en
Inventor
Robert A. Love
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GLO Pharma Inc
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GLO Pharma Inc
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Priority to EP22776585.6A priority Critical patent/EP4313100A4/en
Priority to JP2023559735A priority patent/JP2024511214A/ja
Priority to MX2023011157A priority patent/MX2023011157A/es
Priority to AU2022242835A priority patent/AU2022242835A1/en
Priority to BR112023019249A priority patent/BR112023019249A2/pt
Priority to CA3213099A priority patent/CA3213099A1/en
Priority to KR1020237036137A priority patent/KR20240035939A/ko
Application filed by GLO Pharma Inc filed Critical GLO Pharma Inc
Publication of WO2022204305A2 publication Critical patent/WO2022204305A2/en
Publication of WO2022204305A3 publication Critical patent/WO2022204305A3/en
Priority to US18/473,073 priority patent/US12162956B2/en
Anticipated expiration legal-status Critical
Priority to US18/907,154 priority patent/US20250026787A1/en
Ceased legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
    • A61K8/14—Liposomes; Vesicles
    • 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
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • 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
    • 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
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
    • C07K1/1072—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups
    • C07K1/1077—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4702—Regulators; Modulating activity
    • C07K14/4703—Inhibitors; Suppressors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/72—Receptors; Cell surface antigens; Cell surface determinants for hormones
    • C07K14/723—G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor

Definitions

  • Skin pigmentation occurs by a pathway involving interactions between keratinocytesin the epidermis, and melanocytes in the basal layer of the epidermis. Exposure to UV light causes keratinocytes to release melanocyte-stimulating hormone (MSH). In humans MSH binds to and activates the melanocortin 1 receptor (MC1R) on the plasma membrane of melanocytes. This initiates the MClR-mediated signaling cascade, which raises melanocyte production of the darkly pigmented melanin known as eumelanin, and increases transfer of melanosomes (small secretable vesicles containing melanin) to keratinocytes. This can result in undesirable skin discoloration, for example, dark spots. There is a need for safe and effective compositions and methods of inhibiting the skin pigmentation pathway to prevent and/or reduce skin discoloration.
  • MSH melanocyte-stimulating hormone
  • the present invention includes compositions and methods relating to novel peptide antagonists that inhibit binding of melanocyte-stimulating hormone (MSH) to the melanocortin 1 receptor (MC1R).
  • the invention includes cosmetic compositions comprising a peptide antagonist of the invention.
  • the peptide antagonists of the invention are useful in topical cosmetic compositions for preventing, reducing, or both, the appearance of skin discoloration due to pigmentation.
  • the invention relates to methods of using a cosmetic composition comprising a peptide antagonist of the invention, e.g., to prevent or reduce skin discoloration due to pigmentation in an individual in need thereof.
  • the invention relates to methods for blocking or inhibiting MSH-MC1R interactions using a composition or method of the invention.
  • a Group A melanocortin 1 receptor (MC1R) peptide antagonist comprising an amino acid sequence: Xaal-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6- Xaa7-Xaa8-Xaa9, wherein: Xaal is absent or selected from: Cys, Met, Sec, Ser, Thr, D-Cys, D-Met, D-Sec, D-Ser, D-Thr, Ala, Gly, Val, Leu, lie, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, Phe, Trp, Tyr, D-Phe, D-Trp, D-Tyr, Asn, Asp, Gin, Glu, D-Asn, D-Asp, D-Gln, D-Glu, and a derivative of Cys, Met, Sec, Ser, Thr, D-Cys, D
  • Xaa8 is selected from: Pro, D-Pro, and a derivative of Pro or D-Pro
  • Xaa9 is selected from: Ala, Gly, Val, Leu, He, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, and a derivative of Ala, Gly, Val, Leu, He, D-Ala, D-Gly, D-Val, D-Leu, or D-Ile; wherein when Xaal is Met, Xaa2 is not Pro; the N-terminus is optionally modified; and the C-terminus is optionally modified.
  • Xaa2 of the Group A peptide antagonist is selected from: D-Pro, Ala,
  • Xaa2 of the Group A peptide antagonist is selected from: Ala, Gly, Val, Leu, He, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, and a derivative of Ala, Gly, Val, Leu, He, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, and a derivative of Ala, Gly, Val, Leu, He, D-Ala, D-Gly, D-Val, D-Leu, or D- Ile.
  • Xaal of the Group A peptide antagonist is absent or selected from: Cys, Sec, Ser, Thr, D-Cys, D-Met, D-Sec, D-Ser, D-Thr, Ala, Gly, Val, Leu, He, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, Phe, Trp, Tyr, D-Phe, D-Trp, D-Tyr, Asn, Asp, Gin, Glu, D- Asn, D-Asp, D-Gln, D-Glu, and a derivative of Cys, Sec, Ser, Thr, D-Cys, D-Met, D-Sec, D- Ser, D-Thr, Ala, Gly, Val, Leu, He, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, Phe, Trp, Tyr, D- Phe, D- Phe, D-Trp, D-Tyr, As
  • Xaal of the Group A peptide antagonist is absent or selected from: D-Cys, D- Met, D-Sec, D-Ser, D-Thr, Ala, Gly, Val, Leu, He, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, Phe, Trp, Tyr, D-Phe, D-Trp, D-Tyr, Asn, Asp, Gin, Glu, D-Asn, D-Asp, D-Gln, D-Glu, and a derivative of D-Cys, D-Met, D-Sec, D-Ser, D-Thr, Ala, Gly, Val, Leu, He, D-Ala, D-Gly, D- Val, D-Leu, D-Ile, Phe, Trp, Tyr, D-Phe, D-Trp, D-Tyr, Asn, Asp, Gin, Glu, D-Asn, D-Asp, D
  • Xaal of the Group A peptide antagonist is absent or selected from: Cys, Sec, D-Cys, D-Met, D-Sec, D-Ser, D-Thr, Gly, Val, He, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, D-Phe, D-Trp, D-Tyr, Asn, Asp, D-Asn, D-Asp, D-Gln, D-Glu, and a derivative of Cys, Sec, D-Cys, D-Met, D-Sec, D-Ser, D-Thr, Gly, Val, He, D-Ala, D-Gly, D- Val, D-Leu, D-Ile, D-Phe, D-Trp, D-Tyr, Asn, Asp, D-Asn, D-Asp, D-Gln, or D-Glu.
  • Xaa2 of the Group A peptide antagonist is selected from: D-Pro, Gly, Val, lie, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, and a derivative of D-Pro, Gly, Val, lie, D-Ala, D-Gly, D-Val, D-Leu, D-Ile.
  • Xaal of the Group A peptide antagonist is absent or selected from: Phe, Trp, Tyr, D-Phe, D-Trp, D-Tyr, and a derivative of Phe, Trp, Tyr, D- Phe, D-Trp, or D-Tyr;
  • Xaa2 is selected from: Ala, Gly, Val, Leu, He, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, and a derivative of Ala, Gly, Val, Leu, He, D-Ala, D-Gly, D-Val, D-Leu, or D- Ile;
  • Xaa3 is selected from: Phe, Trp, Tyr, D-Phe, D-Trp, D-Tyr, and a derivative of Phe, Trp, Tyr, D-Phe, D-Trp, or D-Tyr;
  • Xaa4 is selected from: Arg, His, Lys, D-Arg
  • a Group B melanocortin 1 receptor (MC1R) peptide antagonist comprising an amino acid sequence Xaal-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6- Xaa7 -Xaa8 -Xaa9 -Xaa 10 -Xaa 11 -Xaa 12-Xaa 13 -Xaa 14 -Xaa 15 -Xaa 16-Xaa 17 -Xaa 18 -Xaa 19- Xaa20-Xaa21-Xaa22-Xaa23-Xaa24-Xaa25-Xaa26, wherein: Xaal is absent or Xaal and Xaal 9 form a linkage Xaal-Xaal9; Xaa2 is absent or selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu,
  • Xaal4 is selected from: Arg, His, Lys, and a derivative of Arg, His, or Lys
  • Xaal 5 is selected from: Cys, Met, Sec, Ser, Thr, and a derivative of Cys, Met, Sec, Ser, Thr
  • Xaal6 is selected from: Ala, Gly, Val, Leu, lie, and a derivative of Ala, Gly, Val, Leu, or lie
  • Xaal8 is absent or selected from: Phe, Trp, Tyr, Cys, Met, Sec, Ser, Thr, and a derivative of Phe, Trp, Tyr, Cys, Met, Sec, Ser, or Thr
  • Xaal9 is absent or selected from: Ala, Gly, Val, Leu, He, Arg,
  • Xaa20 is absent or selected from: Arg, His, Lys, and a derivative of Arg, His, or Lys;
  • Xaa21 is absent or selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He;
  • Xaa22 is absent or selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He;
  • Xaa23 is absent or selected from: Cys, Met, Sec, Ser, Thr, and a
  • Xaal of the Group B antagonist is absent; Xaa2 is absent; Xaa3 is selected from: Asn, Asp, Gin, Glu, and a derivative of Asn, Asp, Gin, or Glu; Xaa4 is selected from: Pro and a derivative of Pro; Xaa5 and Xaa26 form a linkage Xaa5-Xaa26; Xaa6 is selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He; Xaa7 is selected from: Cys, Met, Sec, Ser, Thr, and a derivative of Cys, Met, Sec, Ser, or Thr; Xaa8 is selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He; Xaa9 is selected from: Phe, Trp, Tyr, and a derivative of Phe, Trp,
  • Xaa20 is selected from: Arg, His, Lys, and a derivative of Arg, His, or Lys
  • Xaa21 is selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He
  • Xaa22 is selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He
  • Xaa23 is selected from: Cys, Met, Sec, Ser, Thr, and a derivative of Cys, Met, Sec, Ser, Thr
  • Xaa24 is selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He
  • Xaa25 is selected from: Asn, Asp,
  • Xaal of the Group B peptide antagonist is absent; Xaa2 is absent; Xaa3 is selected from: Asn, Asp, Gin, Glu, and a derivative of Asn, Asp, Gin, or Glu; Xaa4 is selected from: Pro and a derivative of Pro; Xaa5 and Xaa26 form a linkage Xaa5-Xaa26; Xaa6 is selected from: Ala, Gly, Val, Leu, lie, and a derivative of Ala, Gly, Val, Leu, or lie; Xaa7 is selected from: Cys, Met, Sec, Ser, Thr, and a derivative of Cys, Met, Sec, Ser, or Thr; Xaa8 is selected from: Ala, Gly, Val, Leu, He, Arg, His, Lys, Asn, Asp, Gin, Glu, and a
  • Xaal of the Group B antagonist is absent; Xaa2 is absent; Xaa3 is selected from: Asn, Asp, Gin, Glu, and a derivative of Asn, Asp, Gin, or Glu; Xaa4 is selected from: Pro and a derivative of Pro; Xaa5 and Xaa26 form a linkage Xaa5 -Xaa26; Xaa6 is selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He; Xaa7 is selected from: Cys, Met, Sec, Ser, Thr, and a derivative of Cys, Met, Sec, Ser, or Thr; Xaa8 and Xaal9 form a linkage Xaa8-Xaal9; Xaa9 is selected from: Phe, Trp, Tyr, and a derivative of Phe, Trp, or Tyr; XaalO and Xaal
  • Xaal and Xaal9 of the Group B antagonist form a linkage Xaal-Xaal9;
  • Xaa2 is selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He;
  • Xaa3 is selected from: Asn, Asp, Gin, Glu, and a derivative of Asn, Asp, Gin, or Glu;
  • Xaa4 is selected from: Pro and a derivative of Pro;
  • Xaa5 and Xaa26 form a linkage Xaa5 -Xaa26;
  • Xaa6 is selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He;
  • Xaa7 selected from: Cys, Met, Sec, Ser, Thr, and a derivative of Cys, Met, Sec, Ser, or Thr;
  • Xaa8 selected from:
  • Xaa24 is selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He; and Xaa25 is selected from: Asn, Asp, Gin, Glu, and a derivative of Asn, Asp, Gin, or Glu.
  • Xaal of the Group B antagonist is absent; XaalO and Xaal7 form a linkage Xaal0-Xaal7;
  • Xaal 1 is selected from: Arg, His, Lys, and a derivative of Arg, His, or Lys;
  • Xaal2 is selected from: Phe, Trp, Tyr, and a derivative of Phe, Trp, or Tyr;
  • Xaal3 is selected from: Phe, Trp, Tyr, and a derivative of Phe, Trp, or Tyr;
  • Xaal4 is selected from: Arg, His, Lys, and a derivative of Arg, His, or Lys;
  • Xaal 5 is selected from: Cys, Met, Sec, Ser, Thr, and a derivative of Cys, Met, Sec, Ser, Thr;
  • Xaal 6 is selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He; and
  • Xaal-Xaa8 of the Group B antagonist are absent; Xaal-Xaa8 are absent; Xaa9 is selected from: Phe, Trp, Tyr, Asn, Asp, Gin, Glu, and a derivative of Phe,
  • Xaal 1 is selected from: Arg, His, Lys, and a derivative of Arg, His, or Lys;
  • Xaal2 is selected from: Phe, Trp, Tyr, and a derivative of Phe, Trp, or Tyr;
  • Xaal3 is selected from: Phe, Trp, Tyr, and a derivative of Phe, Trp, or Tyr;
  • Xaal4 is selected from: Arg, His, Lys, and a derivative of Arg, His, or Lys;
  • Xaal5 is selected from: Cys, Met, Sec, Ser, Thr, and a derivative of Cys, Met, Sec, Ser, Thr;
  • Xaal6 is selected from: Ala, Gly, Val, Leu, lie, and a derivative of Ala, Gly, Val, Leu,
  • Xaal-Xaa7 of the Group B antagonist are absent; Xaa8 and Xaal9 form a linkage Xaa8-Xaal9; Xaa9 is selected from: Phe, Trp, Tyr, and a derivative of Phe, Trp, or Tyr; XaalO and Xaal7 form a linkage Xaal0-Xaal7; Xaal 1 is selected from: Arg, His, Lys, and a derivative of Arg, His, or Lys; Xaal2 is selected from: Phe, Trp, Tyr, and a derivative of Phe, Trp, or Tyr; Xaal3 is selected from: Phe, Trp, Tyr, and a derivative of Phe, Trp, or Tyr; Xaal4 is selected from: Arg, His, Lys, and a derivative of Arg, His, or Lys; Xaal5 is selected from: Cys, Met, Sec, Ser, Thr, and a derivative
  • XaalO and Xaal7 form a linkage Xaal0-Xaal7;
  • Xaal l is selected from: Arg, His, Lys, and a derivative of Arg, His, or Lys;
  • Xaal2 is selected from: Phe, Trp, Tyr, and a derivative of Phe, Trp, or Tyr;
  • Xaal3 is selected from: Phe, Trp, Tyr, and a derivative of Phe, Trp, or Tyr;
  • Xaal4 is selected from: Arg, His, Lys, and a derivative of Arg, His, or Lys;
  • Xaal 5 is selected from: Cys, Met, Sec, Ser, Thr, and a derivative of Cys, Met, Sec, Ser, Thr;
  • Xaal 6 is selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He;
  • Xaal8 is absent or selected from
  • a melanocortin 1 receptor peptide antagonist comprising an amino acid sequence set forth as any of SEQ ID NOS: 4-90.
  • the N-terminal and C-terminal modifications are as shown for these peptides in Tables 1 and 2.
  • the N-terminal and C-terminal modifications are selected from any known in the art or described herein.
  • an MC1R peptide antagonist wherein the MC1R peptide antagonist amino acid sequence consists of a Group A or Group B MC1R peptide antagonist amino acid sequence as described herein, including but not limited to any one of SEQ ID NOS: 4-90.
  • an MC1R peptide antagonist wherein the MC1R peptide antagonist amino acid sequence consists of an isolated Group A or Group B MC1R peptide antagonist amino acid sequence as described herein, including but not limited to any one of SEQ ID NOS: 4-90.
  • each linkage in an MC1R peptide antagonist is independently selected from: a Cys-Cys linkage, a Sec-Sec linkage, a cystathionine linkage, a lactam bridge, a Gly-Gly linkage, a thioether linkage, a dicarba linkage; and a diproline linkage selected from Pro— Pro, D-Pro— D-Pro, D-Pro— Pro, and Pro— D-Pro, in the amino to carboxy -terminal direction.
  • a linkage has a spatial separation between the alpha carbons or the geometric center of each amino acid residue of about 3.5 to about 10 angstroms, wherein the linkage has a spatial separation between the alpha carbons or the geometric centers of each of residue of about 3.5 to about 10 angstroms, or both.
  • the N-terminus of an MC1R peptide antagonist is modified to Ci-C 6 acyl, Ci-C 8 alkyl, C 6 -Ci 2 aralkyl, C5-C10 aryl, C 4 -C 8 heteroaryl, formyl, or a lipid.
  • the C- terminus of an MC1R peptide antagonist is modified to comprise NH 2 , amino-acyl, amino-Ci- C 8 alkyl, amino-C 6 -Ci 2 -aralkyl, amino-C 5 -Cio aryl, amino-C 4 -C 8 heteroaryl, or 0-(Ci-C 8 alkyl).
  • the N-terminus is not modified with an amino acid or a derivative of an amino acid
  • the C-terminus is not modified with an amino acid or a derivative of an amino acid.
  • a lipid is covalently attached to a cysteine, serine, lysine, threonine or tyrosine of the MC1R peptide antagonist.
  • the lipid comprises a C 6 -C 20 alkyl, C 6 -C 20 alkenyl, C 6 -C 20 alkynyl, or C 6 -C 20 acyl group.
  • the lipid comprises a geranyl, farnesyl, or geranylgeranyl group.
  • the lipid comprises a undecyloyl, lauroyl, tridecyloyl, myristoyl, palmitoyl, or stearoyl group. In some embodiments, the lipid is a covalent modification of Cys added by palmitoylation or prenylation.
  • the MC1R receptor peptide antagonist comprises at least one derivative that is a non-canonical amino acid selected from the group consisting of: an aromatic side chain amino acid; a non-aromatic side chain amino acid; an aliphatic side chain amino acid; a side chain amide amino acid; a side chain ester amino acid; a heteroaromatic side chain amino acid; a side chain thiol amino acid; a beta amino acid; and a backbone-modified amino acid.
  • the aromatic side chain amino acid is a derivative of tyrosine, histidine, tryptophan, or phenylalanine.
  • the non-aromatic side chain amino acid is a derivative of serine, threonine, cysteine, methionine, arginine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, proline, glycine, alanine, valine, isoleucine, or leucine.
  • the at least one derivative is a non- canonical amino acid selected from the group consisting of: 2-aminoadipic acid; 3- aminoadipic acid; beta-alanine; beta-aminoproprionic acid; 2-aminobutyric acid; 4- aminobutyric acid; piperidinic acid; 6-aminocaproic acid; 2-aminoheptanoic acid; 2- aminoisobutyric acid; 3-aminoisobutyric acid; 2-aminopimelic acid; 2,4-diaminobutyric acid; desmosine; 2,2'-diaminopimelic acid; 2,3-diaminoproprionic acid; N-ethylglycine; N- ethylasparagine; hydroxylysine; allo-hydroxylysine; 3-hydroxyproline; 4-hydroxyproline; isodesmosine; allo-isoleucine; N-methylglycine; sarcosine; n-methylisoleucine; 6-
  • the melanocortin 1 receptor peptide antagonist inhibits a melanocortin 1 receptor.
  • the inhibition is selective.
  • the inhibition comprises blocking MSH binding to the melanocortin 1 receptor.
  • the IC50 is: about 1 millimolar to about 1 picomolar, less than about200 nM, less than about 150 nM, less than about 100 nM, less than about 75 nM, less than about 50 nM, or less than about 25 nM.
  • the melanocortin 1 receptor peptide antagonist inhibits a melanocortin 1 receptor with an increased activity as compared to the activity of a control.
  • the melanocortin 1 receptor peptide antagonist is a Group A MC1R peptide antagonist, and the control is a peptide having the amino acid sequence of SEQ ID NO: 1.
  • the melanocortin 1 receptor peptide antagonist has an amino acid sequence of any one of SEQ ID NOS: 4-43, and the control is a peptide having the amino acid sequence set forth as SEQ ID NO: 1.
  • the melanocortin 1 receptor peptide antagonist is a Group B MC1R peptide antagonist, and the control is a peptide having the amino acid sequence set forth as any one of SEQ ID NOS: 2, 3, 91, 92, 93 and 94.
  • the melanocortin 1 receptor peptide antagonist has an amino acid sequence of any one of SEQ ID NOS: 44-90, and the control is a peptide having the amino acid sequence set forth as any one of SEQ ID NOS: 2, 3, 91, 92, 93 and 94.
  • the melanocortin 1 receptor peptide antagonist is a Group A or Group B MC1R peptide antagonist, and the control is a peptide having the amino acid sequence set forth as any one of SEQ ID NOS: 1, 2, 3, 91, 92, 93 and 94.
  • the melanocortin 1 receptor peptide antagonist inhibits the melanocortin 1 receptor at an IC 50 of about 0.4 to about 0.8 times the control IC 50. In some embodiments, the melanocortin 1 receptor peptide antagonist activity is greater than the activity of the control by about 1.25 -fold to about 5 -fold. In some embodiments, the activity of the melanocortin 1 receptor peptide antagonist and the activity of the control are determined using the same assay method, in the same experiment, or both.
  • a composition comprising an MC1R peptide antagonist as described herein.
  • the composition is a cosmetic composition for prevention, reduction, and/or improvement of the appearance of skin discoloration in an individual in need thereof.
  • the cosmetic composition is formulated for topical use.
  • the skin discoloration can be caused by pigmentation.
  • the skin discoloration can be caused by hyperpigmentation.
  • the skin discoloration e.g., pigmentation or hyperpigmentation, comprises melanin hyperpigmentation, chloasma, melasma, age spots, freckles, or a combination thereof.
  • a method for preventing or improving skin discoloration in an individual comprising applying an effective amount of cosmetic composition comprising the MC1R peptide antagonist to the skin of the individual.
  • cosmetic composition comprises more than one active ingredient.
  • the cosmetic composition comprises more than oneMClR peptide antagonist of the present disclosure.
  • the cosmetic composition comprises one or more excipient selected from the group consisting of: water, a buffer, an absorption enhancer, a stability enhancer, diaminobutyroyl benzylamide, diacetate, glycerin, a gum, a hydrophilic colloid or derivative, a cellulosic derivative, an emulsifier, a fatty alcohol, an acrylic derivative, a mineral, a surfactant, a fat, an oil, a preservative, a monosaccharide, a disaccharide, a polysaccharide, a glycosaminoglycan, and a chelating agent.
  • excipient selected from the group consisting of: water, a buffer, an absorption enhancer, a stability enhancer, diaminobutyroyl benzylamide, diacetate, glycerin, a gum, a hydrophilic colloid or derivative, a cellulosic derivative, an emulsifier, a fatty
  • the absorption enhancer is selected from the group consisting of: a liposome delivery system, a transfersome delivery system, an ethosome delivery system, a short chain alcohol, a long chain alcohol, a polyalcohol, urea, an amino acid, an amino acid ester, an amine, an amide, l-dodecylazacycloheptan-2-one (AZONE ® ), a derivative of 1- dodecylazacycloheptan-2-one, a pyrrolidone, a pyrrolidone derivative, aterpene, a terpene derivative, a fatty acid, a fatty acid ester, a macrocyclic compound, atenside, a sulfoxide, lecithin vesicles, water surfactants, a polyol, a small molecule tri, tetra, penta, hexa, septa or octa peptide, isoceteth-20, e
  • the stability enhancer is a small molecule peptide.
  • the small molecule peptide is selected from the group consisting of: a tripeptide, a tetrapeptide, a pentapeptide, a hexapeptide, a septapeptide, an octapeptide, Acetyl Hexapeptide-3 Cosmetic Topical Peptide, Melanotan II, ACVR2B (ACE-031),
  • the cosmetic composition further comprises one or more other active ingredient.
  • the one or more other active ingredient is selected from the group consisting of: a second, different, MC1R peptide antagonist, an antioxidant, a retinoid, a growth factor, a collagen stimulating peptide, a carrier peptide, a peptide that inhibits tTAT-superoxide dismutase, a peptide that inhibits a proteinase, a peptide that stimulates hyaluronan synthase 2, and a keratin-based peptide.
  • the cosmetic composition comprises a liposome delivery system.
  • the cosmetic composition is a cream, balm, gel, solution, serum, cosmetic, liquid, lotion, ointment, emulsion, milk, spray, mask, or the like. In some embodiments, the cosmetic composition comprises about 0.01% to about 5% w/w of the MC1R receptor peptide antagonist.
  • a cosmetic composition comp rising the melanocortin 1 receptor peptide antagonist for use in preventing or improving skin discoloration in an individual.
  • the invention includes a method for preventing or temporarily improving the appearance in an individual of skin discoloration caused by pigmentation, comprising applying an effective amount of a cosmetic composition comprising the MC1R receptor peptide antagonist of the disclosure to the individual.
  • the MC 1R receptor peptide antagonist is applied in one or more doses of the cosmetic composition.
  • a single dose of the cosmetic composition is applied about once per hour to about once per 2 weeks.
  • a single dose of the cosmetic composition is applied about once per day.
  • the cosmetic composition can be applied indefinitely with no adverse effect.
  • the dose of the cosmetic composition is a subimmunological dose.
  • a lipid vesicle composition comprising: (a) lipid vesicles each comprising a lipid bilayer comprising vesicle forming lipids, (b) an oil-in-water emulsion entrapped in the lipid vesicles, and stabilized by one or more surfactants; (c) a peptide antagonist of a melanocortin 1 receptor entrapped in the lipid bilayer and/or the oil -in-water emulsion.
  • the peptide antagonist of a melanocortin 1 receptor comprises an amino acid sequence: Xaal-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-Xaa9 wherein: Xaal is absent or selected from: Cys, Met, Sec, Ser, Thr, D-Cys, D-Met, D-Sec, D-Ser, D-Thr, Ala, Gly, Val, Leu, lie, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, Phe, Trp, Tyr, D-Phe, D-Trp, D-Tyr, Asn,
  • Xaa2 is selected from: Pro, D-Pro, Ala, Gly, Val, Leu, lie, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, Phe, Trp, Tyr, D-Phe, D-Trp, D-Tyr, Asn, Asp, Gin, Glu, D-Asn, D-Asp, D-Gln, orD-Glu;
  • Xaa2 is selected from: Pro, D-Pro, Ala, Gly, Val, Leu, lie, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, and a derivative of Pro, D-Pro, Ala, Gly, Val, Leu, He, D-Ala, D-Gly, D-
  • the peptide antagonist of a melanocortin 1 receptor comprises an amino acid: Xaal-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-Xaa9-Xaal0-Xaall-Xaal2-Xaal3-Xaal4- Xaa 15 -Xaa 16-Xaal 7-Xaa 18 -Xaa 19 -Xaa20 -Xaa21 -Xaa22-Xaa23-Xaa24-Xaa25 -Xaa26 wherein: Xaal is absent or Xaal and Xaal 9 form a linkage Xaal -Xaal 9; Xaa2 is absent or selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He; Xaa3 is absent or selected from: Asn, Asp, Gin
  • Xaa7 is absent or selected from: Cys, Met, Sec, Ser, Thr, and a derivative of Cys, Met, Sec, Ser, or Thr
  • Xaa8 is absent or selected from: Ala, Gly, Val, Leu, He, Arg, His, Lys, Asn, Asp, Gin, Glu, and a derivative of Ala, Gly, Val, Leu, He, Arg, His, Lys, Asn, Asp, Gin, or Glu, and/or Xaa8 and Xaa 19 form a linkage Xaa8-Xaal9
  • Xaa9 is absent or selected from: Phe, Trp, Tyr, Asn, Asp, Gin, Glu, and a derivative of Phe, Trp, Tyr, Asn, Asp, Gin, or Glu
  • XaalO and Xaal7 form a linkage Xaal0-Xaal7
  • Xaal 1 is selected from: Arg, His,
  • Xaal 8 is absent or selected from: Phe, Trp, Tyr, Cys, Met, Sec, Ser, Thr, and a derivative of Phe, Trp, Tyr, Cys, Met, Sec,
  • Xaa21 is absent or selected from: Ala, Gly, Val, Leu, lie, and a derivative of Ala, Gly, Val, Leu, or He;
  • Xaa22 is absent or selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly,
  • the peptide antagonist of a melanocortin 1 receptor comprises an amino acid sequence as set forth as any one of SEQ ID NOS: 4-90. In some embodiments, the peptide antagonist of a melanocortin 1 receptor consists of an amino acid sequence of any one of the amino acid sequences described herein. In some embodiments, the peptide antagonist is present at a concentration of from about 0.1 mg/mL to about 10 mg/mL. In some embodiments, the lipid vesicle composition further comprises one or more penetration enhancing agents. In some embodiments, the one or more penetration enhancing agents comprises a non -ionic surfactant or a combination of non -ionic surfactants.
  • the non-ionic surfactant or combination of non-ionic surfactants is selected from polyethylene glycol ethers of fatty alcohols, sorbitan esters, polysorbates, sorbitan esters and polyethylene glycol fatty acid esters and combinations thereof.
  • the polyethylene glycol ethers of fatty alcohols comprise a C 8 -C 2 2 fatty alcohol and a polyethylene glycol group having from about 2 to about 8 ethylene glycol subunits.
  • the polyethylene glycol ethers of fatty alcohols comprise diethyleneglycol hexadecyl ether, 2-(2-octadecoxyethoxy)ethanol, diethyleneglycol monooleyl ether, polyoxyethylene (2) oleyl ether, polyoxyethylene (3) oleyl ether, or polyoxyethylene (5) oleyl ether, or any combination thereof.
  • the sorbitan esters comprise sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan monooleate, sorbitan trioleate, sorbitan sesquioleate, or sorbitan isostearate, or any combinations thereof.
  • the polyethylene glycol fatty acid ester comprises PEG-8 dilaurate, PEG-4 dilaurate, PEG-4 laurate, PEG-8 dioleate, PEG-8 distearate, PEG-8 distearate, PEG-7 glyceryl cocoate, and PEG-20 almond glycerides, or any combination thereof.
  • the polysorbate comprises polysorbate 20, polysorbate 21, polysorbate 40, polysorbate 60, polysorbate 80, polysorbate 85, or any combination thereof.
  • each of the non-ionic surfactants has a hydrophobic-lipophilic balance (HLB) of about 10 or less.
  • the non-ionic surfactant or combination of non-ionic surfactants is present in an amount of from about 0.5 % to about to about 10 % (w/w) of the composition.
  • at least one non-ionic surfactant is present in the oil-in-water emulsion.
  • at least one non-ionic surfactant is present in the lipid bilayer.
  • the one or more penetration enhancing agents comprises a combination of a sorbitan ester, a polysorbate, and a polyethylene glycol fatty acid esters. In some embodiments, the one or more penetration enhancing agents comprises a combination of a polyethylene glycol ether of a fatty alcohol, a sorbitan esters, and a polysorbate. In some embodiments, the one or more penetration enhancing agents comprises monolauroyllysine or dipalmitoyllysine, or a combination thereof.
  • the vesicle forming lipids comprise phospholipids, gly colipids, lecithins, ceramides, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, cardiolipin, phosphatidic acid, cerebroside, or any combination thereof.
  • the vesicle forming lipids comprise phospholipids.
  • the composition comprises vesicle forming lipids in an amount of from about 0.5 % to about 25 % (w/w) of the composition.
  • the composition comprises a cationic surfactant.
  • the cationic surfactant is a mono-cationic surfactant.
  • the cationic surfactant comprises an amino acid and a fatty acid.
  • the amino acid comprises lysine, arginine, or histidine.
  • the cationic surfactant is present in an amount of from about 1 % to about 20 % (w/w) of the composition.
  • the oil-in-water emulsion comprises a triglyceride in the oil component.
  • the triglyceride comprises a medium-chain triglyceride.
  • the triglyceride is present in an amount of from about 1 % to about 35 % (w/w) of the composition.
  • the composition comprises a sterol. In some embodiments, the sterol is present in an amount of from about 1 % to about 5 % (w/w) of the composition.
  • the composition comprises propylene glycol. In some embodiments, the propylene glycol is present in an amount of from about 1 % to about 25 % (w/w) of the composition. In some embodiments, the composition comprises one or more viscosity enhancing agents.
  • the one or more viscosity enhancing agents are present in an amount of from about 0.5 % to about 10 % (w/w) of the composition.
  • the composition further comprises one or more additional agents.
  • the additional agents comprise one or more of a thickener, a preservative, a moisturizer, an emollient, a humectant, an antimicrobial, or any combination thereof.
  • the composition is formulated for topical application to the skin of a subject. In some embodiments, the composition is formulated to deliver the peptide antagonist to a specified layer of the skin of a subject.
  • the composition is formulated as a cream, a lotion, a suspension, or an emulsion.
  • a method of preparing a lipid vesicle composition comprising: a) preparing an oil-in-water emulsion comprising the peptide antagonist of a melanocortin 1 receptor, by mixing oil components of the oil-in-water emulsion with aqueous components of the oil-in-water emulsion, wherein the aqueous components comprise the peptide antagonist of a melanocortin 1 receptor, wherein the oil components and/or the aqueous components of the oil -in-water emulsion comprises the one or more surfactants; b) solubilizing vesicle forming lipids in an acceptable solvent other than water; c) adding the oil -in water emulsion to the solubilized vesicle forming lipids; and d) mixing the
  • lipid vesicle composition comprises: (a) lipid vesicles each comprising a lipid bilayer comprising vesicle forming lipids, (b) an oil-in-water emulsion entrapped in the lipid vesicles, and stabilized by one or more surfactants; (c) a peptide antagonist of a melanocortin 1 receptor entrapped in the lipid bilayer and/or the oil-in-water emulsion.
  • the skin discoloration is melanin hyperpigmentation, chloasma, melasma, age spots, freckles, or a combination thereof.
  • FIG. 1 shows an exemplary pictorial workflow for the preparation of lipid vesicles provided herein.
  • FIG. 2 shows an exemplary workflow for the preparation of lipid vesicles comprising a peptide as provided herein.
  • FIG. 3 shows an exemplary result of a subject from a clinical study.
  • the term “comprise” or variations thereof such as “comprises” or “comprising” are to be read to indicate the inclusion of any recited feature but not the exclusion of any other features.
  • the term “comprising” is inclusive and does not exclude additional, unrecited features.
  • “comprising” may be replaced with “consisting essentially of or “consisting of. " The phrase “consisting essentially of is u sed herein to require the specified feature(s) as well as those which do not materially affect the character or function of the claimed invention.
  • the term “consisting” is used to indicate the presence of the recited feature alone.
  • Amino refers to the -NH2 radical.
  • Cyano refers to the -CN radical.
  • Niro refers to the -N0 2 radical.
  • Oxa refers to the -O- radical.
  • Alkyl refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to fifteen carbon atoms (e.g., C1-C15 alkyl).
  • an alkyl comprises one to thirteen carbon atoms (e.g., C1-C13 alkyl).
  • an alkyl comprises one to eight carbon atoms (e.g., Ci-Cg alkyl).
  • an alkyl comprises one to five carbon atoms (e.g., C 1 -C 5 alkyl).
  • an alkyl comprises one to four carbon atoms (e.g., C 1 -C 4 alkyl). In other embodiments, an alkyl comprises one to three carbon atoms (e.g., C 1 -C 3 alkyl). In other embodiments, an alkyl comprises one to two carbon atoms (e.g., Ci-C 2 alkyl). In other embodiments, an alkyl comprises one carbon atom ( e.g ., Ci alkyl). In other embodiments, an alkyl comprises five to fifteen carbon atoms ( e.g ., C 5 -C 15 alkyl). In other embodiments, an alkyl comprises five to eight carbon atoms (e.g., C 5 - C 8 alkyl).
  • an alkyl comprises two to five carbon atoms (e.g., C 2 -C 5 alkyl). In other embodiments, an alkyl comprises three to five carbon atoms (e.g., C 3 -C 5 alkyl).
  • the alkyl group is selected from methyl, ethyl, 1 -propyl (n- propyl), 1 -methylethyl (iso-propyl), 1 -butyl (n -butyl), 1 -methylpropyl (sec-butyl), 2- methylpropyl (iso-butyl), 1 , 1 -dimethylethyl (tent-butyl), 1 -pentyl (n-pentyl).
  • the alkyl is attached to the rest of the molecule by a single bond.
  • an alkyl group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, -OR a , -SR a - 0C(0)-R a , -N(R a ) 2 , -C(0)R a , -C(0)0R a , C(0)N(R a ) 2 , -N(R a )C(0)0R a , -0C(0)-N(R a ) 2 , - N(R a )C(0)R a , -N(R a )C(0) t R a (where t is 1 or 2), -S(0) t 0R a (where t is 1 or 2), -S(0) t R a (where t is 1 or 2) and -S(0) t N(
  • Alkoxy refers to a radical bonded through an oxygen atom of the formula -O-alkyl, where alkyl is an alkyl chain as defined above.
  • Alkenyl refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one carbon -carbon double bond, and having from two to twelve carbon atoms. In certain embodiments, an alkenyl comprises two to eight carbon atoms. In other embodiments, an alkenyl comprises two to four carbon atoms. The alkenyl is attached to the rest of the molecule by a single bond, for example, ethenyl (i.e., vinyl), prop-l-enyl (i.e., allyl), but-l-enyl, pent-l-enyl, penta-l,4-dienyl, and the like.
  • ethenyl i.e., vinyl
  • prop-l-enyl i.e., allyl
  • but-l-enyl pent-l-enyl, penta-l,4-dienyl, and the like.
  • an alkenyl group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, -OR a , -SR a -OC(0)-R a , -N(R a ) 2 , -
  • Alkynyl refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one carbon -carbon triple bond, having from two to twelve carbon atoms.
  • an alkynyl comprises two to eight carbon atoms.
  • an alkynyl comprises two to six carbon atoms.
  • an alkynyl comprises two to four carbon atoms.
  • the alkynyl is attached to the rest of the molecule by a single bond, for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like.
  • an alkynyl group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, -OR a , -SR a -OC(0)-R a , -N(R a ) 2 , -C(0)R a , - C(0)OR a , C(0)N(R a ) 2 , -N (R a )C(0)OR a , -OC(0)-N(R a ) 2 , -N(R a )C(0)R a , -N(R a )C(0) t R a (where t is 1 or 2), -S(0) t OR a (where t is 1 or 2), -S(0) t R a (where t is 1 or 2) and -S(0) t N(R a )
  • Alkylene or "alkylene chain” refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing no unsaturation and having from one to twelve carbon atoms, for example, methylene, ethylene, propylene, n-butylene, and the like.
  • the alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond.
  • the points of attachment of the alkylene chain to the rest of the molecule and to the radical group are through one carbon in the alkylene chain or through any two carbons within the chain.
  • an alkylene comprises one to eight carbon atoms (e.g., Ci-Cg alkylene). In other embodiments, an alkylene comprises one to five carbon atoms
  • an alkylene comprises one to four carbon atoms
  • an alkylene comprises one to three carbon atoms
  • an alkylene comprises one to two carbon atoms
  • an alkylene comprises one carbon atom (e.g., Ci alkylene). In other embodiments, an alkylene comprises five to eight carbon atoms (e.g., C 5 -C 8 alkylene). In other embodiments, an alkylene comprises two to five carbon atoms (e.g., C 2 -C 5 alkylene). In other embodiments, an alkylene comprises three to five carbon atoms (e.g., C3- C 5 alkylene).
  • an alkylene chain is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, -OR a , -SR a -0C(0)-R a , -N(R a ) 2 , -C(0)R a , -C(0)0R a , C(0)N(R a ) 2 , -N(R a )C(0)0R a , -0C(0)-N(R a ) 2 , -N(R a )C(0)R a , -N(R a )C(0) t R a (where t is 1 or 2), -S(0) t 0R a (where t is 1 or 2), -S(0) t R a (where t is 1 or 2) and -S(0) t N(R
  • Aryl refers to a radical derived from an aromatic monocyclic or multicyclic hydrocarbon ring system by removing a hydrogen atom from a ring carbon atom.
  • the aromatic monocyclic or multicyclic hydrocarbon ring system contains only hydrogen and carbon from five to eighteen carbon atoms, where at least one of the rings in the ring system is fully unsaturated, i.e., it contains a cyclic, delocalized (4n+2) 7r-electron system in accordance with the Hiickel theory.
  • the ring system from which aryl groups are derived include, but are not limited to, groups such as benzene, fluorene, indane, indene, tetralin and naphthalene.
  • aryl or the prefix “ar-” (such as in “aralkyl”) is meantto include aryl radicals optionally substituted by one or more substituents independently selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -R b -OR a , -R b -0C(0)- R a , -R b -0C(0)-0R a , -R b -0C(0)-N(R a
  • Aralkyl refers to a radical of the formula -R c -aryl where R c is an alkylene chain as defined above, for example, methylene, ethylene, and the like.
  • the alkylene chain part of the aralkyl radical is optionally substituted as described above for an alkylene chain.
  • the aryl part of the aralkyl radical is optionally substituted as described above for an aryl group.
  • Carbocyclyl or “cycloalkyl” refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which includes fused or bridged ring systems, having from three to fifteen carbon atoms.
  • a carbocyclyl comprises three to ten carbon atoms.
  • a carbocyclyl comprises five to seven carbon atoms.
  • the carbocyclyl is attached to the rest of the molecule by a single bond. Carbocyclyl is saturated (i.e., containing single C-C bonds only) or unsaturated (i.e., containing one or more double bonds or triple bonds).
  • a fully saturated carbocyclyl radical is also referred to as "cycloalkyl.”
  • monocyclic cycloalkyls include, e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
  • An unsaturated carbocyclyl is also referred to as "cycloalkenyl.”
  • Examples of monocyclic cycloalkenyls include, e.g., cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl.
  • Polycyclic carbocyclyl radicals include, for example, adamantyl, norbornyl (i.e., bicyclo[2.2.1]heptanyl), norbomenyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like.
  • carbocyclyl is meant to include carbocyclyl radicals that are optionally substituted by one or more substituents independently selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, - R b -OR a , -R b -0C(0)-R a , -R b -0C(0)-0R a , -R b -0C(0)-N(R a
  • Carbocyclylalkyl refers to a radical of the formula -R c -carbocyclyl where R c is an alkylene chain as defined above.
  • R c is an alkylene chain as defined above.
  • the alkylene chain and the carbocyclyl radical are optionally substituted as defined above.
  • Halo or "halogen” refers to bromo, chloro, fluoro or iodo substituents.
  • Fluoroalkyl refers to an alkyl radical, as defined above, that is substituted by one or more fluoro radicals, as defined above, for example, trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, l-fluoromethyl-2-fluoroethyl, and the like.
  • the alkyl part of the fluoroalkyl radical is optionally substituted as defined above for an alkyl group.
  • Heterocyclyl or “heterocycloalkyl” refers to a stable 3- to 18-membered non aromatic ring radical that comprises two to twelve carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sulfur. Unless stated otherwise specifically in the specification, the heterocyclyl radical is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which optionally includes fused or bridged ring systems. The heteroatoms in the heterocyclyl radical are optionally oxidized. One or more nitrogen atoms, if present, are optionally quaternized. The heterocyclyl radical is partially or fully saturated.
  • heterocyclyl is attached to the rest of the molecule through any atom of the ring(s).
  • heterocyclyl radicals include, but are not limited to, dioxolanyl, thienyl[l,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2- oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydro
  • heterocyclyl is meant to include heterocyclyl radicals as defined above that are optionally substituted by one or more substituents selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl , - R b -OR a , -R b -OC(0)-R a , -R b -0C(0)-0R a , -R b -OC(0)-N(R a ) 2 ,
  • N-heterocyclyl or “N-attachedheterocyclyl” refers to a heterocyclyl radical as defined above containing at least one nitrogen and where the point of attachment of the heterocyclyl radical to the rest of the molecule is through a nitrogen atom in the heterocyclyl radical.
  • An N- heterocyclyl radical is optionally substituted as described above for heterocyclyl radicals. Examples of such N-heterocyclyl radicals include, but are not limited to, 1 -morpholinyl, 1- piperidinyl, 1-piperazinyl, 1 -pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl.
  • C-heterocyclyl or “C-attached heterocyclyl” refers to a heterocyclyl radical as defined above containing at least one heteroatom and where the point of attachment of the heterocyclyl radical to the rest of the molecule is through a carbon atom in the heterocyclyl radical.
  • a C- heterocyclyl radical is optionally substituted as described above for heterocyclyl radicals. Examples of such C-heterocyclyl radicals include, but are not limited to, 2 -morpholinyl, 2- or 3- or 4-piperidinyl, 2-piperazinyl, 2- or 3 -pyrrolidinyl, and the like.
  • Heteroaryl refers to a radical derived from a 3 - to 18-membered aromatic ring radical that comprises two to seventeen carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sulfur.
  • the heteroaryl radical is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, wherein at least one of the rings in the ring system is fully unsaturated, i.e., it contains a cyclic, delocalized (4n+2) p- electron system in accordance with the Hiickel theory.
  • Heteroaryl includes fused or bridged ring systems.
  • the heteroatom(s) in the heteroaryl radical is optionally oxidized.
  • heteroaryl is attached to the rest of the molecule through any atom of the ring(s).
  • heteroaryls include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzooxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][l,4]dioxepinyl, benzo[b][l,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benz
  • heteroaryl is meant to include heteroaryl radicals as defined above which are optionally substituted by one or more substituents selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, haloalkenyl, haloalkynyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, - R b -OR a , -R b -OC(0)-R a , -R b -OC(0)-OR
  • N-heteroaryl refers to a heteroaryl radical as defined above containing at least one nitrogen and where the point of attachment of the heteroaryl radical to the rest of the molecule is through a nitrogen atom in the heteroaryl radical.
  • An N-heteroaryl radical is optionally substituted as described above for heteroaryl radicals.
  • C-heteroaryl refers to a heteroaryl radical as defined above and where the point of attachment of the heteroaryl radical to the rest of the molecule is through a carbon atom in the heteroaryl radical.
  • a C-heteroaryl radical is optionally substituted as described above for heteroaryl radicals.
  • the compounds disclosed herein in some embodiments, contain one or more asymmetric centers and thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that are defined in terms of absolute stereochemistry, as (R)- or (S)-. Unless stated otherwise, it is intended that all stereoisomeric forms of the compounds disclosed herein are contemplated by this disclosure. When the compounds described herein contain alkene double bonds, and unless specified otherwise, it is intended that this disclosure includes both E andZ geometric isomers (e.g., cis or trans.) Likewise, all possible isomers, as well as their racemic and optically pure forms, and all tautomeric forms are also intended to be included.
  • geometric isomer refers to E or Z geometric isomers (e.g., cis or trans) of an alkene double bond.
  • positional isomer refers to structural isomers around a central ring, such as ortho-, meta-, and para- isomers around a benzene ring.
  • a "tautomer” refers to a molecule wherein a proton shift from one atom of a molecule to another atom of the same molecule is possible.
  • the compounds disclosed herein are used in different enriched isotopic forms, e.g., enriched in the content of 2 H, 3 H, 13 C, and/or 14 C.
  • the compound is deuterated in at least one position.
  • deuterated forms can be made by the procedure described in U.S. Patent Nos. 5,846,514 and 6,334,997.
  • deuteration can improve the metabolic stability and or efficacy, thus increasing the duration of action of drugs.
  • structures depicted herein are intended to include compounds which differ only in the presence of one or more isotopically enriched atoms.
  • compounds having the present structures except for the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by 13 C- or 14 C-enriched carbon are within the scope of the present disclosure.
  • the compounds of the present disclosure optionally contain unnatural proportions of atomic isotopes at one or more atoms that constitute such compounds.
  • the compounds may be labeled with isotopes, such as for example, deuterium ( 2 H), tritium ( 3 H), iodine-125 ( 125 I) or carbon-14 ( 14 C) Isotopic substitution with 2 H U C 13 C 14 C 15 C 12 N 13 N 15 N 16 N 16 0 17 0 14 F 15 F 16 F 1T F 18 F 33 S , 34 S 35 S , 36 S , 35 C1, 37 C1, 79 Br, 81 Br, 125 I are all contemplated. All isotopic variations of the compounds of the present disclosure, whether radioactive or not, are encompassed within the scope of the present disclosure.
  • the compounds disclosed herein have some or all of the ⁇ atoms replaced with 2 H atoms.
  • the methods of synthesis for deuterium -containing compounds are known in the art and include, by way of non-limiting example only, the following synthetic methods.
  • Deuterium substituted compounds are synthesized using various methods such as described in: Dean, Dennis C.; Editor. Recent Advances in the Synthesis and Applications of Radiolabeled Compounds for Drug Discovery and Development. [In: Curr., Pharm. Des., 2000; 6(10)] 2000, 110 pp; George W.; Varma, Raj ender S. The Synthesis of Radiolabeled Compounds via Organometallic Intermediates, Tetrahedron, 1989, 45(21), 6601 -21; and Evans, E. Anthony. Synthesis of radiolabeled compounds, J. Radioanal. Chem., 1981, 64(1 - 2), 9-32.
  • Deuterated starting materials are readily available and are subjected to the synthetic methods described herein to provideforthe synthesis of deuterium -containing compounds.
  • Large numbers of deuterium-containing reagents and building blocks are available commercially from chemical vendors, such as Aldrich Chemical Co.
  • “Pharmaceutically acceptable salt” includes both acid and base addition salts.
  • a pharmaceutically acceptable salt of any one of the compounds described herein is intended to encompass any and all pharmaceutically suitable salt forms.
  • Preferred pharmaceutically acceptable salts of the compounds described herein are pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. In certain pharmaceutical embodiments of the present disclosure a “pharmaceutically acceptable salt” may be utilized.
  • “Pharmaceutically acceptable acid addition salt” refers to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid, hydrofluoric acid, phosphorous acid, and the like. Also included are salts that are formed with organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and. aromatic sulfonic acids, etc.
  • acetic acid trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonicacid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.
  • Exemplary salts thus include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, trifluoroacetates, propionates, caprylates, isobutyrates, oxalates, malonates, succinate sub erates, sebacates, fumarates, maleates, mandelates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, phthalates,benzenesulfonates, toluenesulfonates, phenylacetates, citrates, lactates, malates, tartrates, methanesulfonates, and the like.
  • Acid addition salts of basic compounds are, in some embodiments, prepared by contactingthe free base forms with a sufficient amount of the desired acid to produce the salt according to methods and techniques with which a skilled artisan is familiar.
  • a “pharmaceutically acceptable acid addition salt” maybe utilized.
  • “Pharmaceutically acceptable base addition salt” refers to those salts that retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from addition of an inorganic base or an organic base to the free acid. Pharmaceutically acceptable base addition salts are, in some embodiments, formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like.
  • Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, for example, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, N,N-dibenzylethylenediamine, chloroprocaine, hydrabamine, choline, betaine, ethylenediamine, ethylenedianiline, N-methylglucamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N- ethylpiperidine, polyamine resins and the like. See Berge et
  • treatment of or “treating,” “applying,” “palliating,” or “ameliorating” are used interchangeably. These terms refer to an approach for obtaining beneficial or desired results including but not limited to therapeutic benefit and/ora prophylactic benefit.
  • therapeutic benefit is meant eradication or amelioration of the underlying disorder or condition being treated. Also, a therapeutic benefit is achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder or condition such that an improvement is observed in the patient, notwithstanding that the patient is still afflicted with the underlying disorder or condition.
  • compositions are, in some embodiments, administered to a patient at risk of developing a particular disorder or condition, or to a patient reporting one or more of the physiological symptoms of a disorder or condition, even though a diagnosis of this disorder or condition has not been made.
  • treatment of or "treating" the terms "treatment of or "treating,”
  • prodrug is meant to indicate a compound that is, in some embodiments, converted under physiological conditions or by solvolysis to a biologically active compound described herein.
  • prodrug refers to a precursor of a biologically active compound that is pharmaceutically acceptable.
  • a prodrug is typically inactive when administered to a subject, but is converted in vivo to an active compound, for example, by hydrolysis.
  • the prodrug compound often offers advantages of solubility, tissue compatibility or delayed release in a mammalian organism (see, e.g., Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam).
  • the terms "prodrug” may be utilized.
  • prodrugs are provided in Higuchi, T., et al., "Pro -drugs as Novel Delivery Systems," A.C.S. Symposium Series, Vol. 14, and in Bioreversible Carriers in DrugDesign, ed. Edward B. Roche, American Pharmaceutical Association andPergamon Press, 1987.
  • prodrug is also meant to include any covalently bonded carriers, which release the active compound in vivo when such prodrug is administered to a mammalian subject.
  • Prodrugs of an active compound, as described herein are prepared by modifying functional groups present in the active compound in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent active compound.
  • Prodrugs include compounds wherein a hydroxy, amino or mercapto group is bonded to any group that, when the prodrug of the active compound is administered to a mammalian subject, cleaves to form a free hydroxy, free amino or free mercapto group, respectively.
  • Examples of prodrugs include, but are not limited to, acetate, formate and benzoate derivatives of alcohol or amine functional groups in the active compounds and the like.
  • “conservative substitution” means an exchange of one amino acid for another amino acid with similar properties, such as size, charge, and polarity.
  • the substitution can be for a natural or modified (e.g., unnatural amino acid).
  • Non-limiting of examples which can be interchanged in conservative substitutions include the following groupings: Large Hydrophobics (Valine, Leucine, Isoleucine, Phenylalanine, Tryptophan, Tyrosine, Methionine), Small Non-Polar (Alanine, Glycine), Polar (Serine, Threonine, Glutamine, Asparagine, Cysteine, histidine), Positively Charged (Lysine, Arginine), andNegatively Charged (Glutamate, Aspartate).
  • the term “penetration enhancing agents” and “penetration enhancers” are used herein interchangeably. As used herein, it refers to one or more ingredients which facilitate or increase the penetration of one or more active ingredients (e.g., anionic polymeric materials such as hyaluronic acid or peptide antagonists) through one or more layers of the skin of a subject.
  • the penetration enhancing agent is a surfactant, including, for example, non-ionic surfactants having a hydrophilic-lipophilic balance (HLB) of about 10 or less, a cationic group, or another agent such as a terpene, alkaloid, salicylate derivative, nicotinate derivative, or any combination thereof.
  • HLB hydrophilic-lipophilic balance
  • multisome refers lipid vesicle (such as a biphasic lipid vesicle) which comprises one or more penetration enhancers, which in preferred embodiments include multiple penetration enhancers which work in a synergistic fashion.
  • multisomes include vesicle whose central core compartments are occupied by an oil-in-water emulsion composed of an aqueous continuous phase and a dispersed hydrophobic, hydrophilic or oil phase.
  • the spaces between adjacent bilayers of lipid vesicles may also be occupied by the emulsion.
  • lipid vesicle composition refers to a composition which includes one or more lipid vesicles (e.g., multisomal lipid vesicles, lipid bilayer vesicles, etc.).
  • lipid vesicle composition is described as “comprising” one or more additional components (e.g., an anionic polymer material or a peptide antagonist provided herein)
  • additional components e.g., an anionic polymer material or a peptide antagonist provided herein
  • the composition includes the additional component in any manner within the composition (e.g., encapsulated within a lipid vesicle.
  • a lipid vesicle composition comprising an anionic polymer material can include the anionic polymer material encapsulated within a lipid bilayer of the lipid vesicle composition.
  • emulsion refers to a mixture of two immiscible substances.
  • bilayer refers to a structure composed of amphiphilic lipid molecules arranged in two molecular layers, with the hydrophobic tails on the interior and the polar head groups on the exterior surfaces.
  • the term “topical administration” or “topical delivery” as used herein means intradermal, transdermal and/or transmucosal delivery of a compound by administration of a composition comprising the compound or compounds to skin and/or a mucosal membrane.
  • gemini surfactant refers to a surfactant molecule which contains more than one hydrophobic tail, and each hydrophobic tail having a hydrophilic head wherein he hydrophobic tails or hydrophilic heads are linked together by a spacer moiety.
  • the hydrophobic tails can be identical or differ.
  • the hydrophilic heads can be identical or differ the hydrophilic heads may be anionic, cationic, or neutral.
  • HLB Hydrophilic-Lipophilic Balance
  • M1R Melanocortin 1 Receptor
  • the present disclosure relates to peptide antagonists of Melanocortin 1 Receptor, a 317 amino acid G protein coupled receptor found on melanocyte plasma membrane, also referred to as
  • MC1R has 7 a-helical transmembrane domains with a DRY (Asp-Arg-Tyr) motif at the junction of the third transmembrane domain, an intracellular C-terminus with a palmitoylation site, and an extracellular N-terminus with anN-linked glycosylation site. Three intracellular loops and three extracellular loops are formed by its insertion in the membrane. The extracellular and transmembrane domains interact with MC1R ligands. Intracellular and transmembrane domains regulate signaling, including adenylyl cyclase interactions. (See, e.g., Wolf Horrell, 2016, Frontiers in Genetics 7(95): 1 -16, incorporated herein by reference.)
  • MC1R binds pituitary hormones including Adrenocorticotropic hormone (ACTH) and alpha-MSH (cr-MSH), which is a non-selective agonist of melanocortin receptors (MCR-1, 3, 4, and 5).
  • ACTH Adrenocorticotropic hormone
  • cr-MSH alpha-MSH
  • MCR-1, 3, 4, and 5 melanocortin receptors
  • the cr-melanocyte-stimulating hormone ( a - MSH) is produced in response to UV radiation damage of epidermal keratinocytes.
  • the a- MSH binds to MC1R on the melanocyte surface, signaling an increase in melanin pigment synthesis.
  • cr-MSH agonism of MC1R raises melanocyte production of eumelanin, which is darkly pigmented, and increases transfer of melanosomes containing eumelanin to keratinocytes. While dark pigment can protect the individual from UV exposure and further damage, excessive pigmentation is often undesirable.
  • the peptide antagonists of the disclosure bind in the active site of MCI R, blocking the binding of cr-MSH to MC1R and ultimately resulting in decreased production of eumelanin and decreased melanin deposition in the epidermis.
  • a peptide antagonist of the disclosure has a desirable property, or an improved property relative to an MC1R antagonist known in the art.
  • a property can include, e.g., a pharmacokinetic property (including but not limited to absorption, bioavailability, distribution, metabolism, and excretion), a pharmacodynamic property (including but not limited to: receptor binding characteristics, e.g., binding half-life; postreceptor effects; and chemical interactions), enhanced activity (e.g., represented by IC 50 ), stability (e.g., represented by half-life), solubility (e.g., in a formulation), or permeability (e.g., permeability of the skin by a formulation containing the peptide antagonist).
  • a pharmacokinetic property including but not limited to absorption, bioavailability, distribution, metabolism, and excretion
  • a pharmacodynamic property including but not limited to: receptor binding characteristics, e.g., binding half-life; postreceptor effects; and chemical interactions
  • a formulation containing a peptide antagonist of the disclosure has a desirable property, or an improved property relative to a formulation containing an MC1R antagonist known in the art.
  • a desirable or improved property of a formulation of the disclosure is a property relating to the use of the formulation for an indication as described elsewhere herein, e.g., use for reducing or improving the appearance of skin discoloration.
  • an MC1R peptide antagonist of the disclosure is a Group A MC1R peptide antagonist, also referred to herein as a Group A peptide antagonist.
  • a Group A MC1R peptide antagonist of the disclosure has 8-9 amino acid residues and comprises or consists of the amino acid sequence:
  • Xaal is absent or selected from: Cys, Met, Sec, Ser, Thr, D-Cys, D-Met, D-Sec, D- Ser, D-Thr, Ala, Gly, Val, Leu, lie, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, Phe, Trp, Tyr, D- Phe, D-Trp, D-Tyr, Asn, Asp, Gin, Glu, D-Asn, D-Asp, D-Gln, D-Glu, and a derivative of Cys, Met, Sec, Ser, Thr, D-Cys, D-Met, D-Sec, D-Ser, D-Thr, Ala, Gly, Val, Leu, lie, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, Phe, Trp, Tyr, D-Phe, D-Trp, D-Tyr, Asn, Asp,
  • Xaa2 is selected from: Pro, D-Pro, Ala, Gly, Val, Leu, He, D-Ala, D-Gly, D-Val, D- Leu, D-Ile, and a derivative of Pro, D-Pro, Ala, Gly, Val, Leu, He, D-Ala, D-Gly, D-Val, D- Leu, or D-Ile;
  • Xaa3 is selected from: Phe, Trp, Tyr, D-Phe, D-Trp, D-Tyr, and a derivative of Phe, Trp, Tyr, D-Phe, D-Trp, or D-Tyr;
  • Xaa4 is selected from: Arg, His, Lys, D-Arg, D-His, D-Lys, and a derivative of Arg, His, Lys, D-Arg, D-His, or D-Lys;
  • Xaa5 is selected from: Phe, Trp, Tyr, D-Phe, D-Trp, D-Tyr, and a derivative of Phe, Trp, Tyr, D-Phe, D-Trp, or D-Tyr;
  • Xaa6 is selected from: Phe, Trp, Tyr, D-Phe, D-Trp, D-Tyr, and a derivative of Phe, Trp, Tyr, D-Phe, D-Trp, or D-Tyr;
  • Xaa7 is selected from: Arg, His, Lys, D-Arg, D-His, D-Lys, and a derivative of Arg, His, Lys, D-Arg, D-His, or D-Lys;
  • Xaa8 is selected from: Pro, D-Pro, and a derivative of Pro or D-Pro;
  • Xaa9 is selected from: Ala, Gly, Val, Leu, lie, D-Ala, D-Gly, D-Val, D-Leu, D-Ile, and a derivative of Ala, Gly, Val, Leu, lie, D-Ala, D-Gly, D-Val, D-Leu, orD-Ile;
  • N-terminus is optionally modified; and [0094] the C-terminus is optionally modified.
  • the peptide antagonist of the disclosure does not consist of the following amino acid sequence:
  • the number of amino acid residues in a Group A peptide antagonist of the disclosure is not more than 8 or not more than 9 In embodiments, a Group A peptide antagonist of the disclosure consists of 8 or 9 amino acid residues.
  • Non-limiting examples of amino acid sequences of Group A peptide antagonists of the disclosure are shown in Table 1.
  • a Group A MC1R peptide antagonist of the disclosure comprises or consists of an amino acid sequence set forth in Table 1.
  • An MC1R peptide antagonist described herein, including those set forth in Table 1 and Table 2 may comprise or consist of the disclosed amino acid sequence having amino and/or carboxy terminal modifications other than those shown.
  • SEQ IDNOs 16 or 32 have a lower IC50 value using a competition assay compared to a reference (e.g., SEQ ID NO 1).
  • the IC 50 value is about 70 nMto about 160 nM.
  • the IC 50 value is about 70 nMto about 75 nM, about 70 nMto about 80 nM, about 70 nMto about 85 nM, about 70 nMto about 90 nM, about 70 nMto about 100 nM, about 70 nMto about 110 nM, about 70 nMto about 120 nM, about70 nMto about 130 nM, about70 nMto about 140 nM, about70 nMto about 150 nM, about 70 nMto about 160 nM, about 75 nMto about 80 nM, about 75 nMto about 85 nM, about 75 nM to about 90 nM, about 75 nM to about 100 nM, about 75 nM to about 110 nM, about75 nMto about 120 nM, about75 nMto about 130 nM, about75 nMto about 140 nM, about 75 nMto about 150 nM, about 75 nM, about 75
  • IC 50 value is about 70 nM, about 75 nM, about 80 nM, about 85 nM, about 90 nM, about 100 nM, about 110 nM, about 120 nM, about 130nM, about 140 nM, about 150 nM, or about 160 nM. In some embodiments, the IC 50 value is at least about 70 nM, about 75 nM, about 80 nM, about 85 nM, about 90 nM, about 100 nM, about 110 nM, about 120nM, about 130 nM, about 140 nM, or about 150 nM.
  • the IC 50 value is at most about 75 nM, about 80 nM, about 85 nM, about 90 nM, about 100 nM, about 110 nM, about 120 nM, about 130 nM, about 140 nM, about 150nM, or about 160nM.
  • an MC1R peptide antagonist of the disclosure is a Group B MC1R peptide antagonist, also referred to herein as a Group B peptide antagonist.
  • a Group B peptide antagonist of the disclosure has 8-26 amino acid residues and comprises the amino acid sequence:
  • Xaal is absent or Xaal and Xaal9 form a linkage Xaal-Xaal9;
  • Xaa2 is absent or selected from: Ala, Gly, Val, Leu, lie, and a derivative of Ala, Gly, Val, Leu, or lie;
  • Xaa3 is absent or selected from: Asn, Asp, Gin, Glu, and a derivative of Asn, Asp,
  • Xaa4 is absent or selected from: Pro and a derivative of Pro;
  • Xaa5 is absent or Xaa5 and Xaa26 form a linkage Xaa5-Xaa26;
  • Xaa6 is absent or selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He;
  • Xaa7 is absent or selected from: Cys, Met, Sec, Ser, Thr, and a derivative of Cys, Met, Sec, Ser, or Thr;
  • Xaa8 is absent or selected from: Ala, Gly, Val, Leu, He, Arg, His, Lys, Asn, Asp, Gin, Glu, and a derivative of Ala, Gly, Val, Leu, He, Arg, His, Lys, Asn, Asp, Gin, or Glu, and/or Xaa8 and Xaal 9 form a linkage Xaa8-Xaal9;
  • Xaa9 is absent or selected from: Phe, Trp, Tyr, Asn, Asp, Gin, Glu, and a derivative of Phe, Trp, Tyr, Asn, Asp, Gin, or Glu;
  • XaalO and Xaal7 form a linkage Xaal0-Xaal7;
  • Xaal l is selected from: Arg, His, Lys, and a derivative of Arg, His, or Lys;
  • Xaal2 is selected from: Phe, Trp, Tyr, and a derivative of Phe, Trp, or Tyr;
  • Xaal3 is selected from: Phe, Trp, Tyr, and a derivative of Phe, Trp, or Tyr;
  • Xaal4 is selected from: Arg, His, Lys, and a derivative of Arg, His, or Lys;
  • Xaal5 is selected from: Cys, Met, Sec, Ser, Thr, and a derivative of Cys, Met, Sec,
  • Xaal6 is selected from: Ala, Gly, Val, Leu, lie, and a derivative of Ala, Gly, Val, Leu, or lie;
  • Xaal8 is absent or selected from: Phe, Trp, Tyr, Cys, Met, Sec, Ser, Thr, and a derivative of Phe, Trp, Tyr, Cys, Met, Sec, Ser, or Thr;
  • Xaal9 is absent or selected from: Ala, Gly, Val, Leu, He, Arg, His, Lys, Asn, Asp,
  • Xaa20 is absent or selected from: Arg, His, Lys, and a derivative of Arg, His, or Lys;
  • Xaa21 is absent or selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He;
  • Xaa22 is absent or selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He;
  • Xaa23 is absent or selected from: Cys, Met, Sec, Ser, Thr, and a derivative of Cys, Met, Sec, Ser, Thr;
  • Xaa24 is absent or selected from: Ala, Gly, Val, Leu, He, and a derivative of Ala, Gly, Val, Leu, or He;
  • Xaa25 is absent or selected from: Asn, Asp, Gin, Glu, and a derivative of Asn, Asp, Gin, or Glu;
  • Xaa26 is absent or Xaa5 and Xaa26 form a linkage Xaa5-Xaa26;
  • the N-terminus is optionally modified; and [0130] the C-terminus is optionally modified.
  • the amino acid sequence of a peptide antagonist of the disclosure does not consist of: Cys-Cys-Asp-Pro-Cys-Ala-Ser-Cys-Gln-Cys-Arg-Phe-Phe- Arg-Ser-Ala-Cys-Ser- Cys-Arg-Val-Leu-Ser-Leu-Asn-Cys (SEQ ID NO: 2) (Agouti Signaling Protein or ASIP-subdomain, residues 107-132 of full-length ASIP (SEQ ID NO: 91)). See, e.g., Wolf Horrell, 2016, cited and incorporated by reference above.
  • the A SIP- sub domain contains linkages between residues 1 and 19, 5 and 26, and 10 and 17.
  • the amino acid sequence of a peptide antagonist of the disclosure does not consist of SEQ ID NO: 2 containing a linkage between residues 1 and 19 (also referred to herein in the context of a Group B peptide as "Xaal-Xaal9" linkage), 5 and 26 (also referred to herein in the context of a Group B peptide as "Xaa5-Xaa26” linkage) and/or 10 and 17 (Group B peptide "Xaal0-Xaal7" linkage).
  • the amino acid sequence of a peptide antagonist of the disclosure does not consist of the Agouti Signaling Protein amino acid sequence set forth as SEQ ID NO: 91. (See, e.g., UniProtKB - P42127, incorporated herein by reference.) In some embodiments, the amino acid sequence of a peptide antagonist of the disclosure does not consist of a portion of SEQ ID NO: 91 having: anN-terminus at any one of positions 1-106; a C-terminus at any one of positions 125-132, orboth.
  • the amino acid sequence of a peptide antagonist of the disclosure does not consist of a portion of SEQ ID NO: 91 having: an N-terminus at any one of positions 1 -106; a C-terminus at any one of positions 125-132, orboth, and containing a linkage between residues 107 and 125, 111 and 132 and/or between residues 116 and 123.
  • the amino acid sequence of a peptide antagonist of the disclosure does not consist of a portion of SEQ ID NO: 91 selected from: 23- 132 (mature ASIP, SEQ ID NO: 92), 80-132, 93-132, and 106-132, each having ornot having a linkage between residues 107 and 125, 111 and 132 and/or 116 and 123.
  • the amino acid sequence of a peptide antagonist of the disclosure does not consist of: Lys-Lys-Val-Val-Arg-Pro-Arg-Thr-Pro-Leu-Ser-Ala-Pro-Cys- Val-Ala-Thr-Arg-Asn-Ser-Cys-Lys-Pro-Pro-Ala-Pro-Ala-Cys-Cys-Asp-Pro-Cys-Ala-Ser-Cys- Tyr-Cys-Arg-Phe-Phe-Arg-Ser-Ala-Cys-Tyr-Cys-Arg-Val-Leu-Ser-Leu-Asn-Cys (ASIP-YY, SEQ ID NO: 93, which is ASIP 80-132, Q115Y, S124Y).
  • ASIP-YY is described by, e.g., McNulty, et al., 2005, J. Mol. Bio.346:1059-1070, incorporated herein by reference in its entirety.
  • the amino acid sequence of a peptide antagonist of the disclosure does not consist of a portion of SEQ ID NO: 93 having an N-terminus at any one of positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,
  • amino acid sequence of a peptide antagonist of the disclosure does not consist of a sequence selected from a portion of SEQ ID NO: 3 having an N-terminus selected from any one of positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,
  • amino acid sequence of a peptide antagonist of the disclosure does not consist of ASIP-YY subdomain residues 28-53 of SEQ ID NO: 94, having or not having a linkage between residues 15 and 33, 19 and 40 and/or 22 and 31.
  • the amino acid sequence of a peptide antagonist of the disclosure does not consist of: Cys-Cys-Asp-Pro-Cys-Ala-Ser-Cys-Tyr-Cys-Arg-Phe-Phe- Arg-Ser-Ala-Cys-Tyr-Cys-Arg-Val-Leu-Ser-Leu-Asn-Cys (SEQ ID NO: 3) (ASIP-YY subdomain, ASIP-YY residues 2853; corresponding to ASIP residues 107-132).
  • the amino acid sequence of a peptide antagonist of the disclosure does not consist of SEQ ID NO: 3 and having or not having a linkage between residues 1 and 19, 5 and 26 and/or 10 and 17.
  • a peptide antagonist of the disclosure does not consist of a sequence selected from a portion of SEQ ID NO: 3 having an N-terminus selected from any one of positions 1, 2, 3, 4, 5, 6, 7, 8, or 9.
  • a peptide antagonist of the disclosure does not consist of a sequence selected from a portion of SEQ ID NO: 3 having an N-terminus selected from any one of positions 1, 2, 3, 4, 5, 6, 7, 8, or 9, and having or not having a linkage between residues 1 and 19, 5 and 26 and/or 10 and 17.
  • the number of amino acid residues in a Group B peptide antagonist of the disclosure is 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or 26. In some embodiments, the number of amino acid residues in a Group B peptide antagonist of the disclosure is not more than 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or 26. In some embodiments, the number of amino acid residues in a Group B peptide antagonist of the disclosure is not more than 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or26.
  • Non-limiting examples of amino acid sequences of Group B peptide antagonists of the disclosure are shown in Table 2.
  • a Group B MCI R peptide antagonist of the disclosure comprises or consists of an amino acid sequence set forth in Table 2.
  • a peptide of the disclosure can comprise L-amino acids, D-amino acids, or a combination thereof.
  • L-amino acids are indicated by no additional designation, e.g., as in "Pro,” or by an upper or lower case L, with or without punctuation, e.g., "L-” as in “L-Pro", “(L)” as in “(L)Pro,” etc.
  • D-amino acids are indicatedby an upper or lower case D, with or without punctuation, e.g., "D-” as in “D-Pro", “(d)” as in “(d)Pro, “d” as in “dPro,” etc.
  • a substitution of an amino acid is made atone or more positions as desired.
  • Amino acids can be classified based on chemical and structural properties of their sidechains, for example, naturally -occurring amino acids can be classified as hydrophobic (norleucine, Met, Ala, Val, Leu, and He), neutral hydrophilic (Cys, Ser, Thr, Asn, and Gin), acidic (Asp and Glu), basic (His, Lys, and Arg), chain orienting (Gly and Pro), and aromatic (Trp, Tyr, and Phe).
  • a conservative amino acid substitution is made by substituting an amino acid of one of the above classes with a different member of that class.
  • conservative substitutions encompass non-naturally occurring amino acid residues, including peptidomimetics and other reversed or inverted forms of amino acid moieties.
  • a non-conservative substitution is made by substituting an amino acid of one of the above classes with a member of a different class.
  • substitution takes into accountthe hydropathic index of an amino acid (see, e.g., Kyte et al., 1982, J. Mol. Biol. 157: 105-131, incorporated herein by reference).
  • the hydropathic profile of a peptide can be calculated by giving each amino acid a numerical value, or hydropathy index, and repetitively averaging these values along the peptide chain.
  • each amino acid is assigned a hydropathic index based on hydrophobicity and charge characteristics.
  • the hydropathic indices used are: isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine/cystine (+2.5); methionine (+1.9); alanine (+1.8); glycine (-0.4); threonine (-0.7); serine (-0.8); tryptophan (-0.9); tyrosine (-1.3); proline (-1.6); histidine (-3.2); glutamate (-3.5); glutamine (- 3.5); aspartate (-3.5); asparagine (-3.5); lysine (-3.9); and arginine (-4.5).
  • an amino acid is substituted with a different amino acid having a hydropathic index within 0.1 to 0.5 of the original amino acid.
  • the hydropathic index is within 0.1, 0.2, 0.3, 0.4, or 0.5 of the original amino acid.
  • amino acid substitutions are be made based on hydrophilicity.
  • the hydrophilicity values used are: arginine (+3.0); lysine (+3.0); aspartate (+3.0.+-.1); glutamate (+3.0.+-.1); serine (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); threonine (-0.4); proline (-0.5.+-.1); alanine (-0.5); histidine (-0.5); cysteine (-1.0); methionine (1.3); valine (-1.5); leucine (-1.8); isoleucine (-1.8); tyrosine (- 2.3); phenylalanine (-2.5) and tryptophan (-3.4).
  • an amino acid is substituted with a different amino acid having a hydrophilicity value within 0.1 to 0.5 of the original amino acid.
  • the hydrophilicity value is within 0.1, 0.2, 0.3, 0.4, or 0.5 ofthe original amino acid.
  • an amino acid is substituted as shown in the table below.
  • an amino acid is replaced with a conservative substitution as set forth in Table 3(1), or a derivative (also referred to as an analog herein) of a conservative substitution.
  • an amino acid is replaced with an alternative substitution as set forth in Table 3(11), showing each full list of alternatives for each amino acid, or a derivative of an alternative substitution.
  • the peptide antagonist of the present disclosure comprises a constraining structure including, but not limited to, a linkage, bridge or any means of ligation between residues at two positions.
  • the peptide is constrained by its ends or at positions within the peptide, or both.
  • the constraining structure influences a peptide antagonist property, e.g., a pharmacokinetic property (including but not limited to absorption, bioavailability, distribution, metabolism, and excretion), a pharmacodynamic property (including but not limited to: receptor binding characteristics, e.g., binding half-life; postreceptor effects; and chemical interactions), enhanced activity (e.g., represented by IC 50 ), stability (e.g., represented by half-life), solubility (e.g., in a formulation), or permeability (e.g., permeability of the skin by a formulation containing the peptide antagonist).
  • the constraining structure enhances stability of the peptide antagonist.
  • the constraining structure enhances permeability through the skin of the peptide antagonist. In certain embodiments, the constraining structure enhances solubility in a formulation, e.g., a topical formulation, of the peptide antagonist.
  • a peptide antagonist that is constrained as described herein is referred to as a macrocyclic peptide or structure.
  • a macrocyclic peptide refers to a closed - ring structure of a linear peptide intramolecularly formed by linkage between two positions in the peptide, referred to as linkage amino acids, linkage amino acid derivatives, linkage molecule, linkage moiety, linkage residue, linkage entity, or the like, as appropriate.
  • linkage amino acids, linkage amino acid derivatives, linkage molecules, linkage moieties, linkage residues, or linkage entities are separated from each other by two or more amino acid residues, bound to each other directly, bound via a linker, or the like.
  • the macrocyclization may be formed by a bond between an N-terminal amino acid and a C-terminal amino acid of a peptide, by a bond between a terminal amino acid and a non-terminal amino acid, or by a bond between non-terminal amino acids.
  • references to a specific amino acid involved in a linkage can use the nomenclature for the unlinked amino acid (e.g., the structure it may have prior to formation of a linkage). It is also understood that certain linkages, e.g., synthetic linkages, may not be formed by connecting two amino acids or derivatives as commonly referenced in the art. Therefore, references to linked amino acids herein may use the most closely approximating language to describe each involved chemical entity at a given residue position in the peptide antagonist.
  • linked entities in the peptide sequence e.g., Group B amino acids Xaa5 and Xaa26, XaalO and Xaal7, Xaal and Xaal9, and Xaa8 and Xaal 9, may be referred to as linked amino acids, regardless of whether they are amino acids as commonly referenced in the art.
  • Xaa5, Xaa26, XaalO, Xaal7, Xaal, Xaal9, Xaa8, and Xaal9 when linked, e.g, to form linkages Xaa5-Xaa26, Xaal0-Xaal7, Xaal-Xaal9, and/or Xaa8-Xaal 9, can be referred to as linked (or linkage -forming) amino acids, linked (or linkage-forming) amino acid derivatives, linked (or linkage -forming) molecules, linked (or linkage-forming) moieties, linked (or linkage -forming) residues, or linked (or linkage forming) entities in the alternative.
  • linkage amino acids can be used to refer to amino acids, molecules, moieties, residues, or entities present at any of Xaa5, Xaa26, XaalO, Xaal 7, Xaal, Xaal 9, Xaa8, and Xaal 9, in the alternative, either when linked or unlinked.
  • two linkage amino acids also can be referred to as linked (or linkage -forming) amino acids, linked (or linkage-forming) amino acid derivatives, linked (or linkage -forming) molecules, linked (or linkage-forming) moieties, linked (or linkage -forming) residues, or linked (or linkage forming) entities in the alternative.
  • linkage amino acids When linked, two linkage amino acids can be referred to as linked (or linkage -forming) amino acids, linked (or linkage-forming) amino acid derivatives, linked (or linkage -forming) molecules, linked (or linkage -forming) moieties, linked (or linkage-forming) residues, or linked (or linkage -forming) entities, in the alternative.
  • two amino acids can be referred to as unlinked (or non-linkage forming) amino acids, unlinked (or non-linkage forming) amino acid derivatives, unlinked molecules, unlinked moieties, unlinked residues, or unlinked entities.
  • each residue at a non-linked amino acid position in a peptide antagonist of the disclosure can be referred to as an amino acid, amino acid derivative, molecule, moiety, residue or entity, or as an unlinked (or non-linkage forming) amino acid, unlinked (or non-linkage forming) amino acid derivative, unlinked (or non- linkage forming) molecule, unlinked (or non-linkage forming) moiety, unlinked (or non linkage forming) residue or unlinked (or non-linkage forming) entity.
  • constraining structure known to those of skill in the art is contemplated for linking the residues.
  • Examples of constraining structures and their respective linkage residues include, but are not limited to linkages or bridges selected from: a disulfide bridge (e.g., a Cys-Cys linkage, wherein each linkage amino acid is a Cys); a Sec-Sec linkage (selenocysteine linkage, wherein each linkage amino acid is a selenocysteine); a cystathionine linkage or bridge (e.g., Ser-Homocysteine linkage), also referred to herein as Cyt-Cyt (e.g., CH 2 -CH 2 -S-CH 2 ); a lactam bridge (e.g., Asp-Lys or Glu-Lys linkage), a thioether linkage (e.g., a lanthionine linkage, including but not limited to Cys — dehydroalan
  • a linkage is selected from: a disulfide bridge having linkage residues Cys-Cys; a selenocysteine linkage having linkage residues Sec-Sec; a cystathionine linkage having linkage residues Ser-Homocysteine; a lactam bridge having residues Asp-Lys or Glu-Lys; a lanthionine linkage having linkage residues Cys — dehydroalanine or a methyl variant, a diproline linkage having linkage residues L-proline, D-proline, or any combination thereof, a diglycine linkage having glycine linkage residues, and a dicarba linkage having linkage residues allyl glycine or prenyl glycine.
  • a linkage amino acid, linkage amino acid derivative, linkage molecule, linkage moiety, linkage residue, or linkage entity is selected from Cys, Sec, Ser, Homocysteine, Asp, Lys, Glu, dehydroalanine, D-Pro, L-Pro, Gly, or an olefin containing amino acid (e.g., allyl glycine or prenyl glycine).
  • the salt bridge forms between Lys and Glu (amino to carboxy terminal direction) or Glu and Lys (amino to carboxy terminal direction).
  • each linkage of a peptide antagonist of the disclosure is independently selected from any linkage including any described in the preceding paragraph, including a linkage selected from: a disulfide bridge formed by two Cys linkage residues, a Sec-Sec linkage formed by two selenocysteine linkage residues, a cystathionine linkage formed by Ser and homocysteine linkage residues, a lactam bridge formed by Asp and Lys linkage residues or Glu and Lys linkage residues, a thioether linkage that is a lanthionine linkage formed by Cys and dehydroalanine or methyl variant residues, a diproline linkage, a diglycine linkage,
  • a peptide antagonist of the disclosure comprises linkages that are the same as one another, or different.
  • a peptide antagonist of the disclosure can be a Group B peptide antagonist comprising one or more linkages between two terminal residues, two non -terminal residues, or between a terminal and a non-terminal residue.
  • a Group B peptide antagonist can comprise linkages (i) Xaa5-Xaa26 and Xaal0-Xaal7, (ii) Xaa5-Xaa26, Xaal0-Xaal7, and Xaa8-Xaal9, (iii) Xaa5-Xaa26, Xaal0-Xaal7, and Xaal-Xaal9, or (iv) Xaal0-Xaal7.
  • a Group B peptide antagonist is 24 amino acids in length and comprises linkages Xaa5-Xaa26 and Xaal0-Xaal7.
  • a Group B peptide antagonist is 24 amino acids in length, comprises linkages at Xaa5-Xaa26 and Xaal0-Xaal7, and Xaa8- Xaal9. In some embodiments, a Group B peptide antagonist is 24 amino acids in length, comprises linkages at Xaa5-Xaa26 and Xaal0-Xaal7, and forms a salt bridge at Xaa8-Xaal9. In some embodiments, the salt bridge forms between Lys and Glu (amino to carboxy terminal direction) or Glu and Lys (amino to carboxy terminal direction).
  • a Group B peptide antagonist is 24 amino acids in length and comprises linkages at Xaa5-Xaa26 and Xaal0-Xaal7, and Xaa8-Xaal9, wherein Xaa8-Xaal9 is a lactam bridge.
  • a Group B peptide antagonist is 24 amino acids in length, comprises disulfide linkages at Xaa5-Xaa26 and Xaal0-Xaal7, and forms a salt bridge or a lactam bridge at Xaa8- Xaal9.
  • a Group B peptide antagonist is 26 amino acids in length and comprises linkages at Xaa5-Xaa26 and Xaal0-Xaal7, and Xaal-Xaal9. In some embodiments, a Group B peptide antagonist is 26 amino acids in length and comprises disulfide linkages at Xaa5-Xaa26 and Xaal0-Xaal7, and Xaal-Xaal9. In some embodiments, a Group B peptide antagonist is 8 amino acids in length and comprises a linkage at XaalO- Xaal7. In some embodiments, a Group B peptide antagonist is 8 amino acids in length and comprises a disulfide linkage at Xaal0-Xaal7.
  • a Group B peptide antagonist is 8 amino acids in length and comprises a diproline linkage at Xaal0-Xaal7. In some embodiments, a Group B peptide antagonist is 10 amino acids in length and comprises a linkage at Xaal0-Xaal7. In some embodiments, a Group B peptide antagonist is 10 amino acids in length and comprises a disulfide linkage at Xaal0-Xaal7. In some embodiments, a Group B peptide antagonist is 12 amino acids in length and comprises linkages at XaalO- Xaal7 and Xaa8-Xaal9, wherein Xaa8 and Xaal9 are N-terminal and C-terminal residues, respectively.
  • a Group B peptide antagonist is 12 amino acids in length and comprises a disulfide linkage at Xaal0-Xaal7 and a linkage between N and C terminal residues Xaa8 and Xaal9. In some embodiments, a Group B peptide antagonist is 12 amino acids in length and comprises a disulfide linkage at Xaal0-Xaal7 and a diglycine linkage between N and C terminal residues Xaa8 and Xaal9. (See, e.g., Knerr et al., 2011, "Synthesis and activity of thioether-containing analogues of the complement inhibitor compstatin," ACS Chem Biol.
  • any appropriate constraining structure resulting from the use of linkage residues as known in the art is contemplated for use in a peptide antagonist of the disclosure.
  • a particular constraining structure is selected based on its resistance to degradation, e.g., degradation causedby the reduction of a disulfidebond constraining structure.
  • the peptide antagonist comprises a constraining structure that resists degradation by reduction.
  • a disulfide bond may be susceptible to degradation and a resulting loss of activity or other desired peptide antagonist property.
  • a cystathione linkage or a linkage of at least two Ci-C 6 heterocycloalkyl rings confers increased stability relative to a disulfide bond.
  • linkages comprise covalent bonds between canonical or non- canonical amino acids such as cystathionine linkages, lactam bridges, diproline bridges, diglycine bridges, or thioether bridges (e.g., a lanthionine linkage).
  • linkages comprise noncovalent bonds between canonical or non-canonical amino acids such as salt bridges.
  • a linkage comprises a dipeptide.
  • a linkage comprises covalent bonds between canonical or non-canonical acid amino acids such as lanthionine or methyllanthionine linkages.
  • a linkage comprises at least one aromatic or non-aromatic ring.
  • a linkage comprises at least one cycloalkyl ring.
  • a linkage comprises at least one heterocyclic ring.
  • a linkage comprises at least two heterocyclic rings.
  • a linkage comprises at least one nitrogen - containing heterocycloalkyl ring.
  • a linkage comprises the structure , wherein A and B are
  • a linkage comprises the structure , wherein
  • a and B are heterocyclic rings.
  • a linkage comprises pyrrolidine, piperidine, dehydropyrrolidine, dehydropiperidine, aziridine, azetidine, oxazolidine, or thiazolidine.
  • a linkage comprises two Ci-C 6 heterocycloalkyl rings.
  • a linkage comprises at least one five-membered heterocycloalkyl ring.
  • a linkage comprises at least one six-membered heterocycloalkyl ring.
  • a linkage comprises two five-membered heterocycloalkyl rings.
  • a linkage comprises at least one non-canonical amino (unnatural) acid residue.
  • a linkage comprises two amino acids (canonical or non-canonical), wherein a first amino acid has the (S) configuration at the alpha position, and the second amino acid has the (R) configuration at the alpha position.
  • a linkage comprises two amino acids (canonical or non-canonical) connected by a peptide bond.
  • a linkage comprises two proline residues (diproline linkage).
  • a linkage comprises two proline residues connected by a peptide bond.
  • a linkage comprises a D -proline and an L-proline (D-proline-L-proline or L-proline-D-proline), two D-prolines, or two L- prolines.
  • the linkage (from N-terminus to C-terminus) comprises L- proline-D-proline. In some embodiments, the linkage (from C-terminus to N-terminus) comprises L-proline-D-proline.
  • a linkage comprises a D-proline and an L-proline, or derivatives thereof. In some embodiments, a linkage comprises an L-proline and a proline derivative. In some embodiments, a linkage comprises an L-proline and a proline derivative, wherein the proline derivative has the R configuration. In some embodiments, a linkage comprises an L-proline and a proline derivative, wherein the proline derivative has the S configuration. In some embodiments, a linkage comprises D-proline-D-proline or derivatives thereof. In some embodiments, a linkage comprises L-proline-L-proline or derivatives thereof.
  • such derivatives comprise substitutions to the pyrrolidine ring of a proline.
  • a linkage comprises a non-canonical amino acid residue selected from 3-fluoroproline, 4-fluoroproline, 3-hydroxyproline, 4-hydroxyproline, 3- aminoproline, 4-aminoproline, 3,4-dehydroproline, aziridine-2-carboxylic acid, azetidine-2- carboxylic acid, pipecolic acid, 4-oxa-proline, 3-thiaproline, or 4-thiaproline.
  • a linkage comprises two amino acids selected from proline, 3-fluoroproline, 4- fluoroproline, 3-hydroxyproline, 4-hydroxyproline, 3-aminoproline, 4-aminoproline, 3,4- dehydroproline, aziridine-2-carboxylic acid, azetidine-2 -carboxylic acid, pipecolic acid, 4- oxa-proline, 3-thiaproline, or 4-thiaproline.
  • a linkage comprises a connection between a C-terminal residue and an N-terminal residue of a peptide to form a macrocyclic structure.
  • the connection between the C-terminal residue and the N-terminal residue of the peptide is a bond between the N-terminal amine of the N-terminal residue and the C-terminal carboxyl group of the C-terminal residue, a bond between a side chain of the N-terminal residue and the C-terminal carboxyl group of the C-terminal residue, a bond between the N-terminal amine of the N-terminal residue and a side chain of the C-terminal residue, or a bond between a side chain of the N-terminal residue and a side chain of the C-terminal residue.
  • the bond between the C-terminal residue and the N-terminal residue of the peptide is a bond between the N-terminal amine of the N-terminal residue and the C-terminal carboxyl group of the C-terminal residue. In some embodiments, the bond between the C- terminal residue and the N-terminal residue is a peptide bond. In some embodiments, both the N-terminal residue and the C-terminal residue are glycine.
  • the connection between the C-terminal residue and the N- terminal residue of the peptide comprises a linker between the C-terminal residue and the N- terminal residue, such as an alkylene or heteroalkylene linker.
  • the linker is attached to the N-terminal amine of the N-terminal residue, the C-terminal carboxyl group of the C-terminal residue, or a side chain of one or both of the N-terminal or C-terminal residue.
  • the linker is a polymeric linker, such as a polyethylene glycol (PEG) linker.
  • the C-terminal residue and the N-terminal residue of the peptide are separated by about2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 consecutive amino acid residues.
  • a linkage comprises covalent bonds between canonical or non - canonical amino acids lactam bridges.
  • a linkage comprises the structure:
  • the linkage comprises covalent bonds between canonical or non-canonical amino acids thioether bridges.
  • a linkage comprises the structure: [0164] These and similar constraining structures can be used to link residues at terminal and/or nonterminal positions in the peptide.
  • Xaa3 and Xaa8 of a peptide antagonist of the disclosure are linked.
  • Xaa4 and Xaal4 of a peptide antagonist of the disclosure are linked.
  • Xaa3 and Xaa8, and Xaa4 and Xaal4, of a peptide antagonist of the disclosure are linked.
  • a constraining structure as described herein is selected based on the resulting spatial separation between the constrained residues.
  • the spatial separation influences a peptide antagonist property as described above.
  • a peptide antagonist of the disclosure can comprise a constraining structure conferring a spatial separation of about 3.5 to about 10 Angstroms between alpha-carbons of the two linked amino acid residues, or between the geometrical centers of the two linked residues (e.g., amino acid derivatives).
  • the spatial separation between the alpha-carbons of the two linked amino acid residues, or the spatial separation between the geometrical centers of the two linked residues is about 3.5 Angstroms to about 10 Angstroms.
  • the spatial separation between the alpha-carbons of the two linked amino acid residues, or the spatial separation between the geometrical centers of the two linked residues is at least about 3.5 Angstroms. In some embodiments, the spatial separation between the alpha-carbons of the two linked amino acid residues, or the spatial separation between the geometrical centers of the two linked residues, is at most about 10 Angstroms.
  • the spatial separation between the alpha-carbons of the two linked amino acid residues, or the spatial separation between the geometrical centers of the two linked residues is about 3.5 Angstroms to about4.5 Angstroms, about 3.5 Angstroms to about 5 Angstroms, about 3.5 Angstroms to about 5.5 Angstroms, about 3.5 Angstroms to about 6 Angstroms, about 3.5 Angstroms to about 6.5 Angstroms, about 3.5 Angstroms to about 7 Angstroms, about 3.5 Angstroms to about 7.5 Angstroms, about 3.5 Angstroms to about 8 Angstroms, about 3.5 Angstroms to about 8.5 Angstroms, about 3.5 Angstroms to about 9 Angstroms, about 3.5 Angstroms to about 10 Angstroms, about 4.5 Angstroms to about 5 Angstroms, about 4.5 Angstroms to about 5.5 Angstroms, about 4.5 Angstroms to about 6 Angstroms, about 4.5 Angstroms to about 6.5 Angstroms, about 4.5 Angstroms to about 7 Angstroms,
  • the spatial separation between the alpha-carbons of the two linked amino acid residues, or the spatial separation between the geometrical centers of the two linked residues is about 3.5 Angstroms, about 4.5 Angstroms, about 5 Angstroms, about 5.5 Angstroms, about 6 Angstroms, about 6.5 Angstroms, about 7 Angstroms, about 7.5 Angstroms, about 8 Angstroms, about 8.5 Angstroms, about 9 Angstroms, or about 10 Angstroms.
  • a specific spatial separation is achieved using a linker or spacer molecule, as known in the art.
  • an amino acid derivative is a non -canonical amino acid.
  • a non-canonical amino acid has an (S) configuration at the alpha position.
  • a non-canonical amino acid has an (R) configuration at the alpha position.
  • a non-canonical amino acid is an alpha amino acid.
  • a non-canonical amino acid is a beta or gamma amino acid.
  • a non-canonical amino acid is selected from the group consisting of: an aromatic side chain amino acid; a non-aromatic side chain amino acid; an aliphatic side chain amino acid; a side chain amide amino acid; a side chain ester amino acid; a heteroaromatic side chain amino acid; a side chain thiol amino acid; a beta amino acid; and a backbone-modified amino acid.
  • a non-canonical amino acid is a derivative of tyrosine, histidine, tryptophan, or phenylalanine.
  • a derivative of an amino acid comprises an ester, amide, disulfide, carbamate, urea, phosphate, ether of the amino acid.
  • a non -aromatic side chain amino acid is a derivative of serine, threonine, cysteine, methionine, arginine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, proline, glycine, alanine, valine, isoleucine, or leucine.
  • a non-canonical amino acid is selected from the group consisting of 2-aminoadipic acid; 3-aminoadipic acid; beta-alanine; beta-aminoproprionic acid; 2-aminobutyric acid; 4-aminobutyric acid; piperidinic acid; 6-aminocaproic acid; 2- aminoheptanoic acid; 2-aminoisobutyric acid; 3-aminoisobutyric acid; 2-aminopimelic acid; 2,4-diaminobutyric acid; desmosine; 2,2'-diaminopimelic acid; 2,3-diaminoproprionic acid; N-ethylglycine; N-ethylasparagine; hydroxylysine; allo-hydroxylysine; 3-hydroxyproline; 4-hydroxyproline; isodesmosine; allo-isoleucine; N-methylglycine; sarcosine; n- methylisoleucine; 6-N-methyl
  • a non-canonical amino acid is a proline derivative.
  • a proline derivative is a hydroxyproline.
  • a proline derivative is 3 -fluoroproline, 4-fluoroproline, 3-hydroxyproline, 4-hydroxyproline, 3- aminoproline, 4-aminoproline, 3,4-dehydroproline, aziridine-2 -carboxylic acid, azetidine-2- carboxylic acid, pipecolic acid, 4-oxa-proline, 3-thiaproline, or 4-thiaproline.
  • a non-canonical amino acid comprises a lipid.
  • a peptide antagonist of the disclosure comprises one or more amino acid derivative or analog, e.g., as known to those of skill in the art and described in the literature or herein.
  • a peptide antagonist of the disclosure comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 1 -2, 1-3, 1-4, 1-5, 1-6, 1-7, 1-8, 1-9, 1-10, 1-11, 1-12, or 1-13 amino acid derivatives.
  • each amino acid derivative present in a peptide antagonist of the disclosure is a non-canonical amino acid independently selected from the group consisting of: an aromatic side chain amino acid; a non-aromatic side chain amino acid; an aliphatic side chain amino acid; a side chain amide amino acid; a side chain ester amino acid; a heteroaromatic side chain amino acid; a side chain thiol amino acid; a beta amino acid; and a backbone-modified amino acid, selected from e.g., the non-canonical amino acids described herein or known in the art and described in the published literature.
  • the peptide antagonist comprises one or more amino acids that have the D-amino acid configuration, and the remaining amino acids in the peptide have the L- amino acid configuration.
  • a non-canonical amino acid is a proline derivative.
  • a proline derivative comprises one or more substitutions on the pyrrolidine ring.
  • a proline derivative comprises one or more substitutions on the pyrrolidine ring, wherein the substitutions comprise halogen, alkoxy, amino, hydroxyl, alkyl (methyl, ethyl), thiol, or alkylthio.
  • a proline derivative comprises one or more substitutions on the pyrrolidine ring, wherein the substitutions comprise halogen, or alkyl (methyl, ethyl).
  • a proline derivative comprises one or more substitutions on the pyrrolidine ring, wherein the substitutions comprise halogen. In some embodiments, a proline derivative comprises one or more substitutions on the pyrrolidine ring, wherein the substitutions comprise alkoxy, hydroxyl, amino. In some embodiments, a proline derivative comprises one or more substitutions on the pyrrolidine ring, wherein the substitutions comprise halogen, alkoxy, alkyl (methyl, ethyl), thiol, or alkylthio.
  • a peptide antagonist of the present disclosure comprises a tracker amino acid derivative that facilitates tracking of the peptide antagonist.
  • Detection of a noncanonical tracker amino acid present in the peptide antagonist during an assay or following administration of a peptide composition to a subject or patient can provide useful information regarding a peptide antagonist property, e.g., a pharmacokinetic property (including but not limited to absorption, bioavailability, distribution, metabolism, and excretion), a pharmacodynamic property (including but not limited to: receptor binding characteristics, e.g., binding half-life; postreceptor effects; and chemical interactions), enhanced activity (e.g., represented by IC50), stability (e.g., represented by half-life), solubility (e.g., in a formulation), or permeability (e.g., permeability of the skin by a formulation containing the peptide antagonist).
  • a pharmacokinetic property including but not limited to absorption, bioavailability, distribution,
  • a peptide comprising a tracker amino acid derivative is detected using any assay appropriate for detecting the particular tracker amino acid derivative present in the peptide antagonist.
  • a tryptophan derivative is the tracker amino acid.
  • the assay comprises a spectroscopic or radiolabeling detection method.
  • the assay measures a pharmacokinetic or pharmacodynamic peptide antagonist property.
  • the N-terminus amino group of the peptide antagonist of the disclosure is modified (N-terminal modifications).
  • the N-terminus of the peptide antagonist is not modified with an additional amino acid or amino acid derivative.
  • an unmodified N terminus comprises hydrogen.
  • an N-terminal modification comprises Ci-Ce acyl, Ci-C 8 alkyl, C1-C12 aralkyl, C 5 -C 10 aryl, C -C 8 heteroaryl, formyl, or a lipid.
  • an N-terminal modification comprises Ci-C 6 aralkyl.
  • an N-terminal modification comprises Ci-C 6 acyl.
  • an N-terminal modification comprises acetyl (Ac). In some embodiments, an N-terminal modification comprises C1-C6 alkyl. In some embodiments, an N-terminal modification comprises methyl, ethyl, propyl, or tert -butyl. In some embodiments, an N-terminal modification comprises Ci-C 6 aralkyl. In some embodiments, an N-terminal modification comprises benzyl. In some embodiments, an N- terminal modification comprises formyl.
  • a peptide described herein e.g., any peptide having an amino acid sequence as listed in Table 1 and Table 2 (irrespective of the N-terminus shown in the table), has any of these N-terminal modifications, or an unmodified N-terminus.
  • the C-terminus acid group of the peptide antagonist of the disclosure is modified (C-terminal modifications).
  • the C-terminus is not modified with an additional amino acid or amino acid derivative.
  • the C- terminus is not modified with a glycine residue.
  • an unmodified C terminus comprises — OH.
  • a C-terminal modification comprises an amino group, wherein the amino group is optionally substituted.
  • a C-terminal modification comprises an amino group, wherein the amino group is unsubstituted (-NH2).
  • a C-terminal modification comprises an amino group, wherein the amino group is substituted.
  • a C-terminal modification comprises -NH2, - amino-acyl, -amino-Ci-C 8 alkyl, -amino-Ci-C6-aralkyl, -amino-Cs-Cioaryl, or -amino-C 4 -C 8 heteroaryl, -amino-C -C 8 heteroaryl, or -0-(Ci-C 8 alkyl).
  • a C-terminal modification comprises -amino-C6-C 12-aralkyl.
  • a C-terminal modification comprises -0-(Ci-C 8 alkyl).
  • a C-terminal modification comprises -amino-C 6 -C 12-aralkyl. In some embodiments, a C-terminal modification comprises -NH-CH 2 Phenyl. In some embodiments, a C-terminal modification comprises -OEt. In some embodiments, a C-terminal modification comprises -OMe. In some embodiments, a peptide described herein, e.g., any peptide having an amino acid sequence as listed in Table 1 and Table 2 (irrespective of the C-terminus shown in the table), has any of these C-terminal modifications or an unmodified C-terminus.
  • both the N-terminus amino group and the C-terminus acid group of the peptide antagonist of the disclosure are modified.
  • a peptide described herein e.g., any peptide having an amino acid sequence as listed in Table 1 and Table 2 (irrespective of the N- and C-termini shown in the table), has N- and C-termini independently selected from any described herein.
  • a peptide described herein e.g., any peptide having an amino acid sequence as listed in Table 1 and Table 2 (irrespective of the N- and C-termini shown in the table), has N- and C-termini independently selected from: Ac, NH 2 , and H.
  • a peptide antagonist of the present disclosure comprises a lipid moiety.
  • the lipid moiety is covalently attached to an amino acid in the peptide.
  • a lipid is attached to the N-terminus.
  • a lipid is attached to a cysteine, serine, lysine, threonine or tyrosine residue of the peptide antagonist (also referred to herein as "cys-lipid,” “ser-lipid,” “lys-lipid,” “thr-lipid,” or “tyr- lipid,” respectively).
  • the lipid is covalently attached to a cysteine or lysine residue of the peptide antagonist.
  • a lipid is attached to a non- canonical amino acid.
  • a lipid comprises a hydrophobic group.
  • a lipid comprises a fatty acid group.
  • a lipid comprises a C 6 -C 20 fatty acid group.
  • a lipid comprises a steroid.
  • a lipid comprises a wax.
  • a lipid comprises an alkyl group.
  • the lipid comprises a C 6 -C 2 o alkyl, C 6 -C 2 o alkenyl, C 6 -C 2 o alkynyl, or C 6 -C 2 o acyl group.
  • the lipid comprises one or more isoprenyl moieties. In some embodiments, the lipid comprises a geranyl, farnesyl, or geranylgeranyl group. In some embodiments, the lipid comprises an undecyloyl, lauroyl, tridecyloyl, myristoyl, palmitoyl, or stearoyl group. In some embodiments, a peptide described herein comprises an ester, amide, or thioester of a fatty acid.
  • the lipid is a covalent modification of cysteine addedby palmitoylation.
  • the lipid addedby palmitoylation is a C 6 -C 2 o alkyl or a palmitoyl group.
  • the lipid is a covalent modification of cysteine added by prenylation.
  • the lipid added by prenylation is a C 6 -C 2 o alkenyl, geranyl, farnesyl, or geranylgeranyl group.
  • a cys-lipid has Structure I or II.
  • a peptide antagonist of the disclosure is assayed to evaluate one ormore peptide properties including but not limited to a pharmacokinetic property (e.g., absorption, bioavailability, distribution, metabolism, excretion, and the like), a pharmacodynamic property (including: receptor binding characteristics, e.g., binding half-life; postreceptor effects; chemical interactions, and the like), enhanced activity (e.g., represented by IC50), stability (e.g., represented by half-life), solubility (e.g., in a formulation), or permeability (e.g., permeability of the skin by a formulation containing the peptide antagonist).
  • a pharmacokinetic property e.g., absorption, bioavailability, distribution, metabolism, excretion, and the like
  • a pharmacodynamic property including: receptor binding characteristics, e.g., binding half-life; postreceptor effects; chemical interactions, and the like
  • enhanced activity e.g., represented by IC
  • an assay is used to measure the inhibition of MC1R binding and/or activation by an MC1R agonist.
  • the agonist is cr-MSH or or b- MSH.
  • an in vivo or in vitro assay known in the art appropriate for testing a peptide antagonist's effect on a G-protein coupled receptor is contemplated for use in evaluating a peptide antagonist of the disclosure.
  • the assay is a competitive binding assay.
  • binding specificity is determined based on the ability of a peptide antagonist to compete with a-MSH for binding to MC1R.
  • the assay is a high-throughput assay.
  • an assay measures the apparent treatment effect in a subject e.g., assessment of a potential reduction or prevention of skin pigmentation or discoloration can be used to evaluate a peptide antagonist.
  • Melanocytes reside in the basal layer of epidermis where they form the epidermal melanin units as a result of the relationship between one melanocyte and 30-40 associated keratinocytes.
  • an assay used herein evaluates the ability of a topically administered MC1R peptide antagonist to reach the melanocytes, in the basal layer of epidermis.
  • Specific exemplary assays for evaluating a peptide antagonist of the disclosure are described in detail herein in the Examples. Any assay known in the art may be used.
  • skin lightening by an MCR1 peptide antagonist is screened in vitro using the MelanoDerm tissue model (MatTek, Inc.), a highly differentiated, three- dimensional tissue culture model of human epidermis containing normal human melanocytes and keratinocytes.
  • MelanoDerm tissue model (MatTek, Inc.), a highly differentiated, three- dimensional tissue culture model of human epidermis containing normal human melanocytes and keratinocytes.
  • MelanoDerm tissue model MelanoDerm tissue model
  • a highly differentiated, three- dimensional tissue culture model of human epidermis containing normal human melanocytes and keratinocytes See, e.g., Makino et al., 2013, J. Drugs Dermatol. 12(3 suppl I):sl6-s20, incorporated herein by reference in its entirety.
  • Pigmentation can be evaluated over the course of 2-3 weeks using a tristimulus chro
  • total melanin content of tissues is also quantified.
  • cultures become increasingly pigmented with retention of normal epithelial morphology with the expected pigmentation level of the donor tissue, i.e., Black>Asian>Caucasian when cultured in media containing alpha-MSH and beta- FGF.
  • negative control cultures become increasingly pigmented while tissues treated topically with cosmetic skin lightening agents containing tyosinase inhibitors such as kojic acid and magnesium ascorbyl phosphate remain distinctly lighter when compared to control cultures.
  • total melanin content has been found to inversely correlate with surface reflectance (L*). Two distinct endpoints, total melanin content and skin color measurement can be used to evaluate skin pigmentation by this method.
  • a property of a peptide antagonist measured is compared with the property of a suitable control.
  • the result can be expressed as a comparison to an appropriate control, e.g., a negative control or a positive control.
  • a negative control may be a non-specific binding protein, or no treatment.
  • a positive control can be any known MC1R antagonist, e.g., Melanostatine-5/Nonapeptide 1 (SEQ ID NO: 1), the 132-amino acid natural MC1R antagonist Agouti Signaling Protein (ASIP) (SEQ ID NO: 91), mature ASIP (SEQ ID NO: 92), a fragment of ASIP, e.g., ASIP 107-132 (SEQ ID NO: 2), ora variant of ASIP or fragment thereof, e.g., ASIP-YY (SEQ ID NO: 93), ASIP-YY 107-132 subdomain (SEQ ID NO: 3), or ASIP-YY 93-132 subdomain (SEQ ID NO: 94).
  • ASIP prevents a-MSH binding to MC1R and subsequent MC1R activation. See, e.g., Wolf Horrell, 2016.
  • a composition comprising an MCR1 peptide antagonist may be applied to one side of a subject's face, and a control applied to the other side.
  • a control may comprise a composition comprising a control as described herein, or a commercial skin lightening product, e.g., a hydroquinone or kojic acid -containing product.
  • This application may be repeated one or more times a day, e.g., in the morning and in the evening.
  • a sunscreen may be applied to the skin to minimize the effect of light exposure during the day.
  • Subjects' skin may be photographed at intervals and evaluated by experts and/or by self-assessment, e.g., by questionnaire.
  • Skin tone may be analyzed by taking measurements on the cheeks and forehead using Multi Probe Adapter (Courage-Khazaka Electronic GmbH, Cologne, Germany) the Mexameter® MX 18, or VISTA® Complexion Analysis system (Can -field Company, Parsippany, NJ, USA).
  • a composition comprising an MCR1 peptide antagonist prevents, reduces, and/or improves the appearance of skin discoloration.
  • the skin discoloration comprises pigmentation or hyperpigmentation. In some embodiments, the skin discoloration comprises brown spots, red spots, sun damage (photodamage) score, ruggedness of skin texture, bloodvessels, and/or any other appropriate parameter understood by those of skill in the art. In some embodiments the reduction of discoloration after about 1 week to about 12 weeks of treatment is about 5% to about 50%. In some embodiments the reduction of discoloration after about 1 week to about 12 weeks of treatment is about 5% to about 50%.
  • the reduction of discoloration after about 1 week to about 12 weeks of treatment is about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 35%, about 5% to about 40%, about 5% to about 45%, about 5% to about 50%, about 10% to about 15%, about 10% to about 20%, about 10% to about 25%, about 10% to about 30%, about 10% to about35%, about 10% to about 40%, about 10% to about 45%, about 10% to about 50%, about 15% to about 20%, about 15% to about 25%, about 15% to about 30%, about 15% to about 35%, about 15% to about 40%, about 15% to about 45%, about 15% to about50%, about20% to about 25%, about20% to about30%, about20% to about35%, about 20% to about 40%, about 20% to about 45%, about 20% to about 50%, about 25% to about 30%, about 25% to about 35%, about 25% to about 40%, about 25% to about 45%, about 25% to about 50%,
  • the reduction of discoloration after about 1 week to about 12 weeks of treatment is about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50%. In some embodiments the reduction of discoloration after about 1 week to about 12 weeks of treatment is at least about 5%, about 10%, about 15%, about20%, about25%, about 30%, about35%, about40%, or about45%. In some embodiments the reduction of discoloration after about 1 week to about 12 weeks of treatment is at most about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50%.
  • the reduction of discoloration is about 5% to 50% after about 1 week of treatment to about 12 weeks of treatment. In some embodiments the reduction of discoloration is about 5% to about 50% after about 1 week of treatment to about 2 weeks of treatment, about 1 week of treatment to about 3 weeks of treatment, about 1 week of treatment to about 4 weeks of treatment, about 1 week of treatment to about 5 weeks of treatment, about 1 week of treatment to about 6 weeks of treatment, about 1 week of treatment to about 7 weeks of treatment, about 1 week of treatment to about 8 weeks of treatment, about 1 week of treatment to about 9 weeks of treatment, about 1 week of treatment to about 10 weeks of treatment, about 1 week of treatment to about 11 weeks of treatment, about 1 week of treatment to about 12 weeks of treatment, about 2 weeks of treatment to about 3 weeks of treatment, about 2 weeks of treatment to about 4 weeks of treatment, about 2 weeks of treatment to about 5 weeks of treatment, about 2 weeks of treatment to about 6 weeks of treatment, about 2 weeks of treatment to about 7 weeks of treatment, about 2 weeks of treatment to about 8 weeks of treatment, about 2 weeks of
  • the reduction of discoloration is about 5% to 50% after about 1 week of treatment, about 2 weeks of treatment, about 3 weeks of treatment, about 4 weeks of treatment, about 5 weeks of treatment, about 6 weeks of treatment, about 7 weeks of treatment, about 8 weeks of treatment, about 9 weeks of treatment, about 10 weeks of treatment, about 11 weeks of treatment, or about 12 weeks of treatment. In some embodiments the reduction of discoloration is about 5% to 50% after at least about 1 week of treatment, about 2 weeks of treatment, about 3 weeks of treatment, about 4 weeks of treatment, about 5 weeks of treatment, about 6 weeks of treatment, about 7 weeks of treatment, about 8 weeks of treatment, about 9 weeks of treatment, about 10 weeks of treatment, or about 11 weeks of treatment.
  • the reduction of discoloration is about 5% to 50% after at most about 2 weeks of treatment, about 3 weeks of treatment, about 4 weeks of treatment, about 5 weeks of treatment, about 6 weeks of treatment, about 7 weeks of treatment, about 8 weeks of treatment, about 9 weeks of treatment, about 10 weeks of treatment, about 11 weeks of treatment, or about 12 weeks of treatment.
  • the reduction of discoloration after about 1 week to about 12 weeks of treatment is about 5% to about 50% in about 50% to about 100% of subjects.
  • the reduction of discoloration after about 1 week to about 12 weeks of treatment is about 5% to about 50% in about 50% of subjects to about 100% of subjects.
  • the reduction of discoloration after about 1 week to about 12 weeks of treatment is about 5% to about 50% in about 50% of subjects to about 55% of subjects, about 50% of subjects to about 60% of subjects, about 50% of subjects to about 65% of subjects, about 50% of subjects to about 70% of subjects, about 50% of subjects to about 75% of subjects, about 50% of subjects to about 80% of subjects, about 50% of subjects to about 85% of subjects, about 50% of subjects to about 90% of subjects, about 50% of subjects to about 95% of subjects, about 50% of subjects to about 100% of subjects, about 55% of subjects to about 60% of subjects, about 55% of subjects to about 65% of subjects, about 55% of subjects to about 70% of subjects, about 55% of subjects to about 75% of subjects, about 55% of subjects to about 80% of subjects, about 55% of subjects to about 85% of subjects, about 55% of subjects to about 90% of subjects, about 55% of subjects to about 95% of subjects, about 55% of subjects to about 100% of subjects, about 60% of subjects to about 65% of subjects, about 60% of subjects to about
  • the reduction of discoloration after about 1 week to about 12 weeks of treatment is about 5% to about 50% in about 50% of subjects, about 55% of subjects, about 60% of subjects, about 65% of subjects, about 70% of subjects, about 75% of subjects, about 80% of subjects, about 85% of subjects, about 90% of subjects, about 95% of subjects, or about 100% of subjects. In some embodiments the reduction of discoloration after about 1 week to about 12 weeks of treatment is about 5% to about 50% in at least about 50% of subjects, about 55% of subjects, about 60% of subjects, about 65% of subjects, about 70% of subjects, about 75% of subjects, about 80% of subjects, about 85% of subjects, about 90% of subjects, or about 95% of subjects.
  • the reduction of discoloration after about 1 week to about 12 weeks of treatment is about 5% to about 50% in at most about 55% of subjects, about 60% of subjects, about 65% of subjects, about 70% of subjects, about 75% of subjects, about 80% of subjects, about 85% of subjects, about 90% of subjects, about 95% of subjects, or about 100% of subjects.
  • a peptide antagonist of the present disclosure can be compared with that of a known antagonist, e.g., Melanostatine-5 (SEQ ID NO: 1), ASIP (SEQ ID NO: 2), or ASIP-YY (SEQ ID NO: 3).
  • a peptide antagonist is compared in an assay to a control.
  • the control is a negative control, e.g., a random peptide.
  • the control is positive control, e.g., any MC1R antagonist known in the art.
  • the MC1R peptide antagonist tested is a Group A peptide antagonist, e.g., having an amino acid sequence of any one of SEQ ID NOS: 4-43, and the control is a peptide having the amino acid sequence set forth as SEQ ID NO: 1.
  • the MC1R peptide antagonist is a Group B peptide antagonist, e.g., having an amino acid sequence of any one of SEQ ID NOS: 44-90
  • the control is a peptide having the amino acid sequence set forth as any one of SEQ ID NOS: 2, 3, and 91 -94.
  • the MC1R peptide antagonist is a Group B peptide antagonist, e.g., having an amino acid sequence of any one of SEQ ID NOS: 44-90, and the control is a peptide having the amino acid sequence set forth as SEQ ID NO: 2.
  • the MC1R peptide antagonist is a Group B peptide antagonist, e.g., having an amino acid sequence of any one of SEQ ID NOS: 44-90, and the control is a peptide having the amino acid sequence set forth as SEQ ID NO: 3.
  • the inhibition of MCI R activation by a peptide antagonist of the present disclosure is compared with that of a known antagonist using the same assay.
  • the inhibition of MCI R activation by a peptide antagonist of the present disclosure is compared with that of a known antagonist in the same experiment. In embodiments, the inhibition of MC1R activation by a peptide antagonist of the present disclosure is compared with that of a known antagonist in the same experiment, using the same assay.
  • a peptide antagonist of the disclosure inhibits MC1R at an IC 50 of about 1 millimolar to about 1 picomolar. In some embodiments, the peptide antagonist has an IC50 of about 1 millimolar to about 1 picomolar in a receptor binding assay.
  • the IC50 is about 1 millimolar to about 1 micromolar, about 1 millimolar to about 500 micromolar, about 500 micromolar to about 100 micromolar, about 100 micromolar to about 1 micromolar, about 1 micromolar to about 1 nanomolar, about 1 micromolar to about 1 nanomolar, about 1 micromolar to about 500 nanomolar, about 500 nanomolar to about 100 nanomolar, about 100 nanomolar to about 1 nanomolar, about 1 nanomolar to about 1 picomolar, about 1 nanomolar to about 500 picomolar, about 500 picomolar to about 100 picomolar, about 100 micromolar to about 1 picomolar, about or less than about 1 millimolar, about or less than about 500 millimolar, about or less than about 100 millimolar, about or less than about 1 millimolar, about or less than about 1 micromolar, about or less than about 500 micromolar, about or less than about 100 micromolar, about or less than about 50 micromolar, about or less than about 20 micromolar
  • the IC 50 is about20 nanomolarto about 125 nanomolar. In some embodiments, the IC50 is about 20 nanomolarto about 25 nanomolar, about 20 nanomolarto about 30 nanomolar, about 20 nanomolar to about 40 nanomolar, about 20 nanomolarto about 50 nanomolar, about 20 nanomolar to about 60 nanomolar, about 20 nanomolarto about 70 nanomolar, about 20 nanomolar to about 75 nanomolar, about 20 nanomolar to about 80 nanomolar, about 20 nanomolar to about 90 nanomolar, ab out 20 nanomolarto about 100 nanomolar, about20 nanomolarto about 125 nanomolar, about25 nanomolarto about 30 nanomolar, about 25 nanomolar to about 40 nanomolar, about 25 nanomolarto about 50 nanomolar, about 25 nanomolar to about 60 nanomolar, about 25 nanomolarto about 70 nanomolar, about 25 nanomolar to about 75 nanomolar, about 25 nanomolarto about 80 nanomolar, about 25 nanomolar
  • the IC 50 is about 20 nanomolar, about 25 nanomolar, about 30 nanomolar, about 40 nanomolar, about 50 nanomolar, about 60 nanomolar, about 70 nanomolar, about 75 nanomolar, about 80 nanomolar, about 90 nanomolar, about 100 nanomolar, or about 125 nanomolar. In some embodiments, the IC 50 is at least about 20 nanomolar, about 25 nanomolar, about 30 nanomolar, about 40 nanomolar, about 50 nanomolar, about 60 nanomolar, about 70 nanomolar, about 75 nanomolar, about 80 nanomolar, about 90 nanomolar, or about 100 nanomolar.
  • the IC 50 is at most about25 nanomolar, about 30 nanomolar, about 40 nanomolar, about 50 nanomolar, about 60 nanomolar, about 70 nanomolar, about 75 nanomolar, about 80 nanomolar, about 90 nanomolar, about 100 nanomolar, or about 125 nanomolar.
  • the IC 50 of the peptide antagonist is about 25 nanomolarto about 150 nanomolar. In some embodiments, the IC 50 of the peptide antagonist is at least about 25 nanomolar. In some embodiments, the IC 50 of the peptide antagonist is at most about 150 nanomolar.
  • the IC 50 of the peptide antagonist is about 25 nanomolarto about 35 nanomolar, about 25 nanomolarto about 40 nanomolar, about 25 nanomolarto about 45 nanomolar, about 25 nanomolarto about 50 nanomolar, about 25 nanomolarto about 60 nanomolar, about 25 nanomolarto about 70 nanomolar, about 25 nanomolarto about 75 nanomolar, about 25 nanomolar to about 100 nanomolar, about 25 nanomolar to about 110 nanomolar, about 25 nanomolar to about 125 nanomolar, about 25 nanomolar to about 150 nanomolar, about 35 nanomolar to about 40 nanomolar, about 35 nanomolarto about 45 nanomolar, about 35 nanomolar to about 50 nanomolar, about35 nanomolarto about 60 nanomolar, about 35 nanomolar to about 70 nanomolar, about 35 nanomolarto about 75 nanomolar, about 35 nanomolar to about 100 nanomolar, about 35 nanomolar to about 110 nanomolar, about 35 nanomolar to about 125 nanomolar, about
  • the IC50 of the peptide antagonist is about 25 nanomolar, about 35 nanomolar, about 40 nanomolar, about 45 nanomolar, about 50 nanomolar, about 60 nanomolar, about 70 nanomolar, about 75 nanomolar, about 100 nanomolar, about 110 nanomolar, about 125 nanomolar, or about 150 nanomolar.
  • the IC 50 is less than about 200 nM, less than about 150 nM, less than about 100 nM, less than about75 nM, less than about50 nM, or less than about25 nM.
  • the IC5o can be determined by any appropriate activity assay as described herein or known in the art.
  • the IC50 represents the concentration of peptide antagonist at which 50% of the MCI R binding to an MC1R agonist is blocked.
  • the IC 50 observed using a peptide antagonist of the disclosure is lower than that of a control (comparison) MC1R peptide antagonist.
  • a control MC1R peptide antagonist can be any MC1R peptide antagonist known in the art, e.g., Melanostatine- 5/Nonapeptide 1 (SEQ ID NO: 1), the 132-amino acid natural MC1R antagonist Agouti Signaling Protein (ASIP) (SEQ ID NO: 91), mature ASIP (SEQ ID NO: 92), a fragment of ASIP, e.g., ASIP 107-132 (SEQ ID NO: 2), or a variant of ASIP or fragment thereof, e.g., ASIP-YY (SEQ ID NO: 93), ASIP-YY 107-132 subdomain (SEQ ID NO: 3), or ASIP-YY 93- 132 subdomain (SEQ ID NO: 94).
  • the IC 50 observed using a peptide antagonist of the disclosure is about 0.4 to about 0.8 times that of the control antagonist. In some embodiments, the IC50 observed using a peptide antagonist of the disclosure is lower that of the control antagonist by about2-fold to about 10-fold, i.e., the IC 50 is about 10% to about 50% that observed with the control antagonist. In some embodiments, the IC50 observed using a peptide antagonist of the disclosure is lower than that of the control antagonist by about 2- fold to about 10-fold.
  • the IC 50 observed using a peptide antagonist of the disclosure is lower that of the control antagonist by about 2 -fold to about 3 -fold, about 2- foldto about 4-fold, about 2-fold to about 5-fold, about 2-fold to about 6-fold, about 2-fold to about 7-fold, about 2-fold to about 8-fold, about 2-fold to about 9-fold, about 2-fold to about 10-fold, about 3 -fold to about 4-fold, about 3 -fold to about 5-fold, about 3 -fold to about 6- fold, about 3-fold to about 7-fold, about 3-fold to about 8-fold, about 3 -fold to about 9-fold, about3-foldto about 10-fold, about4-fold to about 5-fold, about4-fold to about6-fold, about 4-fold to about7-fold, about 4-fold to about 8-fold, about 4-fold to about 9-fold, about4-fold to about 10-fold, about5-fold to about 6-fold, about5-fold to about7-fold, about 5-foldto about 8-fold, about 5-fold to about
  • the IC 50 observed using a peptide antagonist of the disclosure is lower than that of the control antagonist by about2-fold, about3-fold, about4-fold, about5-fold, about 6-fold, about 7-fold, about 8-fold, about 9-fold, or about 10-fold. In some embodiments, the IC50 observed using a peptide antagonist of the disclosure is lower than that of the control antagonist by at least about 2-fold, about 3 -fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about 8-fold, or about 9-fold.
  • the IC 50 observed using a peptide antagonist of the disclosure is lower than that of the control antagonist by at most about 3-fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about8-fold, about 9- fold, or about 10-fold.
  • the activity of a peptide antagonist of the disclosure activity is greater than the activity of a control by about 1.25 -fold to about 5-fold.
  • the activity of a peptide antagonist of the disclosure activity is greater than the activity of a control by about 1.25 -fold to about 5-fold.
  • the activity of a peptide antagonist of the disclosure activity is greater than the activity of a control by about 1.25-fold to about 1.5-fold, about 1.25-fold to about 1.75-fold, about 1.25- fold to about2-fold, about 1.25-fold to about 2.5 -fold, about 1.25-fold to about3-fold, about
  • the activity of a peptide antagonist of the disclosure activity is greater than the activity of a control by about
  • the activity of a peptide antagonist of the disclosure activity is greater than the activity of a control by at least about 1.25-fold, about 1.5-fold, about 1.75-fold, about2-fold, about 2.5-fold, about3-fold, about 3.5 -fold, about4-fold, or about 4.5 -fold. In some embodiments, the activity of a peptide antagonist of the disclosure activity is greater than the activity of a control by at most about
  • the IC 50 observed for a Group A peptide antagonist of the disclosure is about 10% to about 50% of the IC 50 observed fora control, e.g., SEQ ID NO: 1.
  • the observed for a Group B peptide antagonist of the disclosure is about 10% to about 50% of the IC50 observed for a control, e.g., SEQ ID NO: 2, 3, or 91-94.
  • the IC 50 observed for a Group A or B peptide antagonist of the disclosure is about 10% to about 50% of the IC50 observed fora control, e.g., SEQ ID NO: 1, 2, 3, or 91-94.
  • the MC1R peptide antagonists of the disclosure are contemplated for cosmetic uses in a subject in need thereof, for indications including but not limited to the prevention, reduction, and/or improvement of the appearance of undesirable skin discoloration.
  • the skin discoloration can be caused by pigmentation.
  • the skin discoloration can be caused by hyperpigmentation.
  • the skin discoloration e.g., pigmentation or hyperpigmentation, comprises melanin hyperpigmentation, post-inflammatory hyperpigmentation, chloasma, melasma, age spots (e.g., liver spots, senile lentigines, solar lentigines, sunspots), freckles, or a combination thereof.
  • the MC1R peptide antagonists of the disclosure can be used to treat skin discoloration resulting from a disorder, e.g., an adrenal disorder.
  • the MC1R peptide antagonists of the disclosure are used to treat skin discoloration caused by Addison's disease, in which MC1R agonists ACTH and MSH are overproduced.
  • the MC1R peptide antagonists of the disclosure are used to treat skin discoloration due to hyperpigmentation of any known cause, e.g., drug use (e.g., calcium antagonists), cyanic melasma, senile melasma, vitiligo, adverse sequelae following sclerotherapy, or postinflammatory or traumatic responses.
  • existing skin discoloration is reduced. In some embodiments, further skin discoloration is prevented. In some embodiments, the skin discoloration is improved in appearance, e.g., due to a change in the texture (ruggedness) of the skin.
  • the subject is a mammal.
  • the mammal is a human.
  • the human subject is a pediatric or adult subject, of any age.
  • peptide antagonists of the disclosure can be provided in a cosmetic or pharmaceutical composition.
  • cosmetic or pharmaceutical compositions of the disclosure are: formulated using excipients or carriers that are nottoxic to keratinous tissue, e.g., skin, and are cosmetically, pharmaceutically and/or dermatologically acceptable; and administered in treatments comprising a subimmunological dose of the composition.
  • a formulation of the disclosure e.g., a cosmetic or pharmaceutical composition
  • administration of a formulation of the disclosure is not expected to result in adverse effects, even when administered repeatedly and often, e.g., as described herein.
  • adverse effects are minor, few, or nonexistent.
  • Adverse effects of topical application can include, e.g., mild to severe skin pain, redness, burning, itching, irritation, or any other side effects commonly associated with topical compositions. More severe effects avoided by formulations of the disclosure can include side effects associated with injectable neurotoxins as described in product labeling, e.g., as referenced herein.
  • the present disclosure includes a cosmetic composition comprising a peptide antagonist of the disclosure.
  • the cosmetic composition is formulated for topical administration.
  • the cosmetic composition is formulated for topical administration as a cream, balm, gel, solution, serum, cosmetic, liquid, lotion, ointment, emulsion, milk, spray, mask, or the like.
  • a topical cosmetic or pharmaceutical composition comprises an excipient or carrier or a suitable combination of two, three, or more excipients or carriers.
  • any appropriate excipient or carrier or combination of multiple excipients and/or carriers is selected from excipients known to those of skill in the art and described in the literature, e.g., those useful in a topical formulation.
  • an excipient or carrier useful in a topical formulation is selected from the group consisting of: an inert excipient or carrier, a buffer, an absorption enhancer (penetrating agent), and a stability enhancer.
  • the inert excipient or carrier is water, isopropyi alcohol, gaseous fluorocarbons, ethyl alcohol, polyvinyl pyrrolidone, propylene glycol, a fragrance, a gel-producing material, stearyl alcohol, stearic acid, spermaceti, sorbitan monooleate, or methylcellulose.
  • the excipient is an inorganic compound.
  • an excipient is a carbohydrate.
  • the excipient is diaminobutyroyl benzylamide, diacetate, glycerin, a gum, a hydrophilic colloid or derivative, a cellulosic derivative, an emulsifier, a fatty alcohol, an acrylic derivative, a mineral, a surfactant, a fat, an oil, a preservative, a monosaccharide, a disaccharide, a polysaccharide, a glycosaminoglycan, or a chelating agent.
  • the absorption enhancer is selected from the group consisting of : a liposome delivery system, atransfersome delivery system, an ethosome delivery system, a short chain alcohol, a long chain alcohol, a polyalcohol, urea, an amino acid, an amino acid ester, an amine, an amide, an azacyclo compound (e.g., 1 -dodecylazacycloheptan-2-one (AZONE®) or a derivative of 1 -dodecylazacycloheptan-2-one) a pyrrolidone, a pyrrolidone derivative, a terpene, aterpene derivative, a fatty acid, a fatty acid ester, a macrocyclic compound, atenside, a sulfoxide, lecithin vesicles, water surfactants, a polyol, a small molecule tri, tetra, penta, hexa, sept
  • the absorption enhancer is selected to have a minimal allergic or irritating effect.
  • the stability enhancer is a small molecule peptide.
  • the small molecule peptide is selected from the group consisting of: a tripeptide, a tetrapeptide, a pentapeptide, a hexapeptide, a septapeptide, an octapeptide,
  • the cosmetic composition further comprises one or more additional active ingredient.
  • the one or more additional active ingredient is selected from the group consisting of: a second, different, MC1R peptide antagonist, another skin -lightening agent, an anti-wrinkle agent, a retinoid, an antioxidant, a growth factor, a collagen stimulating peptide, a carrier peptide, a peptide that inhibits tTAT- superoxide dismutase, a peptide that inhibits a proteinase, a peptide that stimulates hyaluronan synthase 2, and a keratin-based peptide.
  • the cosmetic composition comprises a liposome delivery system.
  • the liposome delivery system includes, e.g., oil-in-water emulsions, micelles, mixed micelles, or liposomes.
  • a colloidal system is a liposome or microsphere.
  • a composition is formulated as a poly(D,L)lactide microspheres.
  • the cosmetic composition is formulated to incorporate features as described below, for use in a pharmaceutical composition.
  • the excipient(s) and/or liposome delivery system are selected using methods known to those of skill in the art to achieve the desired degree of penetration, e.g., to achieve substantially local delivery. In some embodiments, the excipient(s) and/or liposome delivery system are selected to target melanocytes in the basal layer of the epidermis.
  • transdermal delivery of an MC1R peptide antagonist described herein is achieved using a liposome or lipid vesicle composition.
  • the lipid vesicle composition comprises lipid vesicles each comprising a lipid bilayer comprising vesicle forming lipids.
  • the lipid vesicle composition comprises an oil - in-water emulsion entrapped in the lipid vesicles.
  • the oil -in-water emulsion is stabilized by one or more surfactants.
  • the MC1R peptide antagonist is entrapped in the lipid bilayer and/or the oil -in-water emulsion. In some embodiments, the MC1R peptide antagonist is entrapped in the lipid bilayer. In some embodiments, the MC1R peptide antagonist is entrapped in the oil-in-water emulsion.
  • the lipid vesicle composition comprises: (a) lipid vesicles each comprising a lipid bilayer comprising vesicle forming lipids; (b) an oil -in-water emulsion entrapped in the lipid vesicles, and stabilized by one or more surfactants; and (c) an MC1R peptide antagonist entrapped in the lipid bilayer and/or the oil -in-water emulsion.
  • the present disclosure also includes pharmaceutical compositions comprising a peptide antagonist of the disclosure.
  • the pharmaceutical composition is formulated as described above for a cosmetic composition comprising a peptide antagonist of the disclosure.
  • a cosmetic or pharmaceutical composition of the disclosure is formulated using any excipient, carrier, or additive as appropriate.
  • compositions comprising one or more MC1R peptide antagonist as described herein, e.g., a Group A or Group B MC1R peptide antagonist having an amino acid sequence as set forth in Table 1 and Table 2, or as otherwise described herein.
  • the composition is a cosmetic or pharmaceutical composition.
  • the disclosure also relates to methods for blocking or inhibiting MSH-MC1R interactions using MC1R peptide antagonist composition, comprising contacting the MC1R peptide antagonist with the MC1R peptide antagonist composition.
  • the disclosure also relates to the use of an MC1R peptide antagonist or composition thereof to prevent or reduce skin discoloration in a subject.
  • the present disclosure also relates to methods for using cosmetic or pharmaceutical compositions comprising a peptide antagonist of the disclosure.
  • the disclosure relates to methods for using the cosmetic or pharmaceutical composition to prevent, reduce, and/or improve the appearance in a subject of skin discoloration, comprising applying an effective amount of the cosmetic or pharmaceutical composition to the skin of the subject.
  • the skin discoloration comprises melanin hyperpigmentation, post- inflammatory hyperpigmentation, chloasma, melasma, age spots (e.g., liver spots, senile lentigines, solar lentigines, sunspots), freckles, or a combination thereof.
  • the skin discoloration results from a disorder, e.g., an adrenal disorder.
  • the MC1R peptide antagonists of the disclosure are used to prevent or reduce skin discoloration caused by Addison's disease.
  • administration is topical.
  • administration is to the skin.
  • administration is topically applied to the skin.
  • a subject in the context of the present disclosure is a mammalian subject.
  • the mammal is a human.
  • the human subject is a pediatric, juvenile, or adult subject, of any age.
  • the subject has a disease or disorder that causes unwanted skin pigmentation.
  • the subject has Addison's disease.
  • the cosmetic or pharmaceutical composition is topically applied to a subject.
  • Topical application as referred to herein can refer to application onto one or more surface, e.g., keratinous tissue. Topical application may relate to direct application to the desired area.
  • a topical cosmetic or pharmaceutical composition or preparation can be applied by, e.g., pouring, dropping, or spraying, when present as a liquid or aerosol composition; smoothing, rubbing, spreading, and the like, when in ointment, lotion, cream, gel, or a like composition; dusting, when a powder; or by any other appropriate means.
  • a subject is treated with an effective amount of the topical cosmetic or pharmaceutical composition during a period between treatments with a skin peeling agent.
  • a subject is treated with a topical cosmetic or pharmaceutical composition of the disclosure periodically beginning on the day after, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26 weeks after a treatment with the skin peeling agent.
  • the subject is treated by topical application of an effective amount of the cosmetic or pharmaceutical composition one time or more during a course of treatment, e.g., 1 -3 times per day, 1-21 times per week, 1 time per day, 2 times per day, or 3 times per day.
  • a subject is treated with an effective amount of the cosmetic or pharmaceutical composition about 1 time per week to about 12 times per week.
  • a subject is treated with an effective amount of the cosmetic or pharmaceutical composition at least about 1 time per week.
  • a subject is treated with an effective amount of the cosmetic or pharmaceutical composition at most about 12 times per week.
  • a subject is treated with an effective amount of the cosmetic or pharmaceutical composition about 1 time per week to about 2 times per week, about 1 time per week to about 3 times per week, about 1 time per week to about 4 times per week, about 1 time per week to about 5 times per week, about 1 time per week to about 6 times per week, about 1 time per week to about 7 times per week, about 1 time per week to about 8 times per week, about 1 time per week to about 9 times per week, about 1 time per week to about 10 times per week, about 1 time per week to about 11 times per week, about 1 time per week to about 12 times per week, about 2 times per week to about 3 times per week, about 2 times per week to about 4 times per week, about 2 times per week to about 5 times per week, about 2 times per week to about 6 times per week, about 2 times per week to about 7 times per week, about 2 times per week to about 8 times per week, about 2 times per week to about 9 times per week, about 2 times per week to about 10 times per week, about 2 times per week to about 2 times per week
  • a subject is treated with an effective amount of the cosmetic or pharmaceutical composition about 1 time per week, about 2 times per week, about 3 times per week, about 4 times per week, about 5 times per week, about 6 times per week, about 7 times per week, about 8 times per week, about 9 times per week, about 10 times per week, about 11 times per week, about 12 times per week, about 13 times per week, or about 14 times per week.
  • a pharmaceutical composition of the disclosure is administered to a subject, for indications including but not limited to: preventing or reducing skin discoloration, comprising applying an effective amount of the cosmetic or pharmaceutical composition to the skin of the subject.
  • the skin discoloration comprises melanin hyperpigmentation, post- inflammatory hyperpigmentation, chloasma, melasma, age spots (e.g., liver spots, senile lentigines, solar lentigines, sunspots), freckles, or a combination thereof.
  • the skin discoloration results from a disorder, e.g., an adrenal disorder.
  • the MC1R peptide antagonists of the disclosure are used to treat skin discoloration caused by Addison's disease.
  • a topical cosmetic composition of the disclosure is self -applied or administered by a patient.
  • a cosmetic or pharmaceutical composition of the disclosure is applied or administered by a medical professional, e.g., in a medical office setting.
  • a lipid vesicle composition comprising a peptide antagonist of a melanocortin 1 receptor, such as those described herein (e.g., SEQ IDNOs: 1-90).
  • the lipid vesicle composition comprises lipid vesicles each comprising a lipid bilayer comprising vesicle forming lipids.
  • the lipid vesicle composition comprises an oil-in-water emulsion entrapped in the lipid vesicles.
  • the oil-in-water emulsion is stabilized by one or more surfactants.
  • the peptide antagonist is entrapped in the lipid bilayer and/or the oil-in-water emulsion. In some embodiments, the peptide antagonist is entrapped in the lipid bilayer. In some embodiments, the peptide antagonist is entrapped in the oil-in-water emulsion.
  • the peptide antagonist is present in the vesicle composition in an amount of about 0.1 mg/mL to about 50 mg/mL. In some embodiments, the peptide antagonist is present in the vesicle composition in an amount of about 0.1 mg/mL to about 0.5 mg/mL, about 0.1 mg/mL to about 1 mg/mL, about 0.1 mg/mL to about 2 mg/mL, about 0.1 mg/mL to about 3 mg/mL, about 0.1 mg/mL to about 4 mg/mL, about 0.1 mg/mL to about 5 mg/mL, about 0.1 mg/mL to about 10 mg/mL, about 0.1 mg/mL to about 20 mg/mL, about 0.1 mg/mL to about 50 mg/mL, about 0.5 mg/mL to about 1 mg/mL, about 0.5 mg/mL to about 2 mg/mL, about 0.5 mg/mL to about 3 mg/mL, about 0.5 mg/mL
  • the peptide antagonist is present in the vesicle composition in an amount of about 0.1 mg/mL, about 0.5 mg/mL, about 1 mg/mL, about 2 mg/mL, about 3 mg/mL, about 4 mg/mL, about 5 mg/mL, about 10 mg/mL, about 20 mg/mL, or about 50 mg/mL. In some embodiments, the peptide antagonist is present in the vesicle composition in an amount of at least about 0.1 mg/mL, about 0.5 mg/mL, about 1 mg/mL, about 2 mg/mL, about 3 mg/mL, about 4 mg/mL, about 5 mg/mL, about 10 mg/mL, or about 20 mg/mL.
  • the peptide antagonist is present in the vesicle composition in an amount of at most about 0.5 mg/mL, about 1 mg/mL, about 2 mg/mL, about 3 mg/mL, about 4 mg/mL, about 5 mg/mL, about 10 mg/mL, about 20 mg/mL, or about 50 mg/mL. In some embodiments, the peptide antagonist is present in the composition in an amount of about 1 mg/mL, about 2 mg/mL, about 3 mg/mL, about 4 mg/mL, or about 5 mg/mL.
  • the vesicle composition comprises one or more vesicle forming lipids.
  • the vesicle forming lipids act to encapsulate portions of the oil -in-water emulsions. In some embodiments, this allows the oil-in-water emulsion to remain stable fora period of time.
  • the vesicle forming lipids may be any suitable lipids for such a purpose.
  • the vesicle forming lipids comprise phospholipids, glycolipids, lecithins, ceramides, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, cardiolipin, phosphatidic acid, cerebroside, or any combination thereof.
  • the vesicle forming lipids comprise a combination of lipids.
  • the vesicle forming lipids comprise phospholipids. .
  • the phospholipids are naturally occurring, semi synthetic, or synthetically prepared, or a mixture thereof.
  • the phospholipids are one or more esters of glycerol with one or two (equal or different) residues of fatty adds and with phosphoric acid, wherein the phosphoric acid residue is in turn bound to a hydrophilic group, such as, for instance, choline (phosphatidylcholines— PC), serine (phosphatidylserines— PS), glycerol (phosphatidylglycerols— PG), ethanolamine (phosphatidylethanolamines— PE), or inositol (phosphatidylinositol).
  • choline phosphatidylcholines— PC
  • serine phosphatidylserines— PS
  • glycerol phosphatidylglycerols— PG
  • ethanolamine phosphati
  • Esters of phospholipids with only one residue of fatty acid are generally referred to in the art as the "lyso" forms of the phospholipid or "lysophospholipids".
  • Fatty acids residues present in the phospholipids are in general long chain aliphatic acids, typically containing 12 to 24 carbon atoms, or 14 to 22 carbon atoms; the aliphatic chain may contain one or more unsaturations or is completely saturated.
  • suitable fatty acids included in the phospholipids are, for instance, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, oleic acid, linoleic acid, and linolenic acid.
  • Saturated fatty acids such as myristic acid, palmitic acid, stearic acid and arachidic acid may be employed.
  • the phospholipid comprises one or more natural phospholipids. In some embodiments, the phospholipid comprises one or more semisynthetic phospholipids. In some embodiments, the semisynthetic phospholipids are the partially or fully hydrogenated derivatives of the naturally occurring lecithins. In some embodiments, the phospholipids include fatty acids di-esters of phosphatidylcholine, ethylphosphatidylcholine, phosphatidylglycerol, phosphatidic acid, phosphatidylethanolamine, phosphatidylserine or of sphingomyelin.
  • the phospholipids include hydrogenated phosphatidylcholine (e.g., Sunlipon 90H).
  • the phospholipids are, for instance, dilauroyl-phosphatidylcholine (DLPC), dimyristoyl-phosphatidylcholine (DMPC), dipalmitoyl-phosphatidylcholine (DPPC), diarachidoyl- phosphatidylcholine (DAPC), distearoyl-phosphatidylcholine (DSPC), dioleoyl-phosphatidylcholine (DOPC), l,2Distearoyl-sn-glycero-3-Ethylphosphocholine (Ethyl-DSPC), dipentadecanoyl- phosphatidylcholine (DPDPC), l-myristoyl-2-palmitoyl-phosphatidylcholine (MPPC), 1- palmitoyl-2- myristoyl-phosphati
  • the vesicle forming lipids are present in an amount of about 0.5 % to about 25 % (w/w) of the composition. In some embodiments, the vesicle forming lipids are present in an amount of about 0.5 % to about 2 %, about 0.5 % to about 5 %, about 0.5 % to about 8 %, about 0.5 % to about 10 %, about 0.5 % to about 12 %, about 0.5 % to about 15 %, about 0.5 % to about 20 %, about 0.5 % to about 25 %, about 2 % to about 5 %, about 2 % to about 8 %, about 2 % to about 10 %, about 2 % to about 12 %, about 2 % to about 15 %, about 2 % to about 20 %, about 2 % to about 25 %, about 5 % to about 8 %, about 5 % to about 10 %, about 5 % to about 10 %, about 5 % to about 12
  • the vesicle forming lipids are present in an amount of about 0.5 %, about 2 %, about 5 %, about8 %, about 10 %, about 12 %, about 15 %, about 20 %, or about25 %. In some embodiments, the vesicle forming lipids are present in an amount of at least about 0.5 %, about 2 %, about 5 %, about 8 %, about 10 %, about 12 %, about 15 %, or about 20 % (w/w) of the composition.
  • the vesicle forming lipids are present in an amount of at most about 2 %, about 5 %, about 8 %, about 10 %, about 12 %, about 15 %, about 20 %, or about 25 % (w/w) of the composition.
  • the vesicle forming lipids are present in an amount of about 5 % to about 15 % (w/w) of the composition. In some embodiments, the vesicle forming lipids are present in an amount of about 5 % to about 8 %, about 5 % to about 9 %, about 5 % to about 10 %, about 5 % to about 11 %, about 5 % to about 12 %, about 5 % to about 13 %, about 5 % to about 14 %, about 5 % to about 15 %, about 8 % to about 9 %, about 8 % to about 10 %, about 8 % to about 11 %, about 8 % to about 12 %, about 8 % to about 13 %, about 8 % to about 14 %, about 8 % to about 15 %, about 9 % to about 10 %, about 9 % to about 11 %, about 9 % to about 12 %, about 8 % to about 13 %, about 8 % to about 14
  • the vesicle forming lipids are present in an amount of about 5 %, about 8 %, about 9 %, about 10 %, about 11 %, about 12 %, about 13 %, about 14 %, or about 15 %(w/w) of the composition. In some embodiments, the vesicle forming lipids are present in an amount of at least about 5 %, about 8 %, about 9 %, about 10 %, about 11 %, about 12 %, about 13 %, or about 14 % (w/w) of the composition.
  • the vesicle forming lipids are present in an amount of at most about 8 %, about 9 %, about 10 %, about 11 %, about 12 %, about 13 %, about 14 %, or about 15 % (w/w) of the composition.
  • the composition comprises a short chain polyol.
  • the short chain polyol acts to enhance the stability of the resulting lipid vesicles.
  • the short chain polyol is a C 2 -C polyol comprising two or three alcohol groups.
  • the short chain polyol is propylene glycol.
  • the composition comprises propylene glycol.
  • the propylene glycol is present in an amount of about 0.5 % to about 25 % (w/w) of the composition. In some embodiments, the propylene glycol is present in an amount of about 0.5 % to about 2 %, about 0.5 % to about 5 %, about 0.5 % to about 8 %, about0.5 % to about 10 %, about0.5 % to about 12 %, about 0.5 % to about 15 %, about 0.5 % to about 20 %, about 0.5 % to about 25 %, about 2 % to about 5 %, about 2 % to about 8 %, about 2 % to about 10 %, about 2 % to about 12 %, about 2 % to about 15 %, about 2 % to about 20 %, about 2 % to about 25 %, about 5 % to about 8 %, about 5 % to about 10 %, about 5 % to about 12 %, about 5 % to about 15 %, about 5 % to about 12 %,
  • the propylene glycol is present in an amount of about 0.5 %, about 2 %, about 5 %, about 8 %, about 10 %, about 12 %, about 15 %, about 20 %, or about 25 %. In some embodiments, the propylene glycol is present in an amount of at least about 0.5 %, about 2 %, about 5 %, about 8 %, about 10 %, about 12 %, about 15 %, or about 20 %. In some embodiments, the propylene glycol is present in an amount of at most about 2 %, about 5 %, about 8 %, about 10 %, about 12 %, about 15 %, about 20 %, or about 25 %.
  • the propylene glycol is present in an amount of about 1 % to about 10 %. In some embodiments, the propylene glycol is present in an amount of about 1 % to about 2 %, about 1 % to about 4 %, about 1 % to about 6 %, about 1 % to about 8 %, about 1 % to about 10 %, about 2 % to about 4 %, about 2 % to about 6 %, about 2 % to about 8 %, about 2 % to about 10 %, about 4 % to about 6 %, about 4 % to about 8 %, about 4 % to about 10 %, about 6 % to about 8 %, about 6 % to about 10 %, or about 8 % to about 10 %.
  • the propylene glycol is present in an amount of about 1 %, about 2 %, about 4 %, about 6 %, about 8 %, or about 10 %. In some embodiments, the propylene glycol is present in an amount of at least about 1 %, about 2 %, about 4 %, about 6 %, or about 8 %. In some embodiments, the propylene glycol is present in an amount of at most about 2 %, about 4 %, about 6 %, about 8 %, or about 10 %. In some embodiments, propylene glycol is present in about the same amount as the vesicle forming lipid. In some embodiments, the ratio of propylene glycol to vesicle forming lipid in the composition is form about2:l to about 1 :2 (w/w).
  • the lipid vesicle compositions provided herein comprise an oil-in-water emulsion.
  • the oil component is selected such that the material is a liquid at operative temperatures (e.g., room temperature) and is non-miscible with water.
  • the oil phase comprises a naturally occurring oil.
  • the naturally occurring oil is derived from one or more plants or plant parts (e.g., seeds or nuts).
  • the oil is a naturally occurring oil such as olive oil, vegetable oil, sunflower oil, or other similar plant derived oil.
  • the oil phase is selected from the group consisting of vegetable oils, mono-, di-, and triglycerides, silicone fluids, mineral oils, and combinations thereof.
  • the oil comprises a silicon oil or derivative, such as dimethicone.
  • the oil silicon oil comprises a siloxane polymer.
  • the siloxane polymer comprises C 1 -C 3 substituents.
  • the siloxane is polydimethylsiloxane (PDMS).
  • the oil is a mixture which comprises a silicon oil (e.g., dimethicone) as a smaller component.
  • the silicon oil is incorporated in order to enhance the feel of the resulting composition or as a moisturizer.
  • the oil comprises a silicon oil in an amount of up to about 5 %, up to about 4%, up to about 3 %, up to about 2%, or up to about 1%.
  • the silicon oil is present in an amount of from about 0.1 % to about 2% (w/w) of the composition. In some embodiments, the silicon oil is present in an amount of about 0.5 %, about 0.6 %, about 0.7 %, about 0.8 %, about 0.9 % or about 1%.
  • the oils are present in an amount of about 1 % to about 35 % (w/w) of the composition. In some embodiments, the oils are present in an amount of about 1 % to about 5 %, about 1 % to about 10 %, about 1 % to about 15 %, about 1 % to about 20 %, about 1 % to about 25 %, about 1 % to about 30 %, about 1 % to about 35 %, about 5 % to about 10 %, about 5 % to about 15 %, about 5 % to about 20 %, about 5 % to about 25 %, about 5 % to about 30 %, about 5 % to about 35 %, about 10 % to about 15 %, about 10 % to about 20 %, about 10 % to about 25 %, about 10 % to about 30 %, about 10 % to about 35 %, about 15 % to about 20 %, about 15 % to about 25 %, about 10 % to about 30 %, about 10 % to
  • the oils are present in an amount of about 1 %, about 5 %, about 10 %, about 15 %, about 20 %, about 25 %, about 30 %, or about 35 %. In some embodiments, the oils are present in an amount of at least about 1 %, about 5 %, about 10 %, about 15 %, about 20 %, about 25 %, or about 30 %. In some embodiments, the oils are present in an amount of at most about 5 %, about 10 %, about 15 %, about 20 %, about 25 %, about 30 %, or about 35 %. In some embodiments, the oils are present in an amount of about 5 % to about 15 %.
  • the oils are present in an amount of about 5 % to about 8 %, about 5 % to about 9 %, about 5 % to about 10 %, about 5 % to about 11 %, about 5 % to about 12 %, about 5 % to about 13 %, about 5 % to about 14 %, about 5 % to about 15 %, about 8 % to about 9 %, about 8 % to about 10 %, about 8 % to about 1 1 %, about 8 % to about 12 %, about 8 % to about 13 %, about 8 % to about 14 %, about 8 % to about 15 %, about 9 % to about 10 %, about 9 % to about 11 %, about 9 % to about 12 %, about 9 % to about 13 %, about 9 % to about 14 %, about 9 % to about 15 %, about 10 % to about 11 %, about 10 % to about 12 %, about 10 % to about 13 %, about 9 % to
  • the oils are present in an amount of about 5 %, about 8 %, about 9 %, about 10 %, about 11 %, about 12 %, about 13 %, about 14 %, or about 15 %. In some embodiments, the oils are present in an amount of at least about 5 %, about 8 %, about 9 %, about 10 %, about 11 %, about 12 %, about 13 %, or about 14 %. In some embodiments, the oils are present in an amount of at most about 8 %, about 9 %, about 10 %, about 11 %, about 12 %, about 13 %, about 14 %, or about 15 %.
  • the oil comprises one or more triglycerides.
  • the triglyceride is a medium chain triglyceride.
  • the medium chain triglyceride comprises fatty acid esters having a chain length of C6-C12.
  • the triglyceride is present in an amount of about 1 % to about 35 % (w/w) of the composition. In some embodiments, the triglyceride is present in an amount of about 1 % to about 5 %, about 1 % to about 10 %, about 1 % to about 15 %, about 1 % to about 20 %, about 1 % to about 25 %, about 1 % to about 30 %, about 1 % to about 35 %, about 5 % to about 10 %, about 5 % to about 15 %, about 5 % to about 20 %, about 5 % to about 25 %, about 5 % to about 30 %, about 5 % to about 35 %, about 10 % to about 15 %, about 10 % to about 20 %, about 10 % to about 25 %, about 10 % to about 30 %, about 10 % to about 35 %, about 15 % to about 20 %, about 10 % to about 25 %, about 10 % to
  • the triglyceride is present in an amount of about 1 %, about 1.5 %, about2 %, about2.5 %, about3 %, about3.5 %, about4 %, about4.5 %, about 5 %, about 10 %, about 15 %, about 20 %, about 25 %, about 30 %, or about 35 %. In some embodiments, the triglyceride is present in an amount of at least about 1 %, about 1.5 %, about 2 %, about 2.5 %, about 3 %, about 3.5 %, about 4 %, about 4.5 %, about 5 %, about 10 %, about 15 %, about 20 %, about 25 %, or about 30 %.
  • the triglyceride is present in an amount of at most about 1.5 %, about 2 %, about 2.5 %, about 3 %, about3.5 %, about4 %, about4.5 %, about 5 %, about 10 %, about 15 %, about 20 %, about 25 %, about 30 %, or about 35 %.
  • the oil phase of the lipid vesicle and/or the lipid vesicle portion of the composition comprises a sterol.
  • the sterol is cholesterol.
  • the cholesterol may be plant-derived cholesterol.
  • the plant-derived cholesterol may be PhytoChol®, SyntheChol®, or any other plant-derived cholesterol (e.g., Av anti# 700100), or any combination thereof.
  • the sterol may be phytosterol or a derivative thereof.
  • the phytosterol or derivative thereof may be Phytosterol MM, AdvasterolTM 90 IP or 95 IP F, NET Sterol-ISO, canola sterols, sitosterol 700095, lanosterol-95, brassicasterol, or any combination thereof.
  • the sterol is present in an amount of about 1 % to about 5 %(w/w) of the composition.
  • the sterol is present in an amount of about 1 % to about 1.5 %, about 1 % to about 2 %, about 1 % to about 2.5 %, about 1 % to about 3 %, about 1 % to about 4 %, about 1 % to about 5 %, about 1.5 % to about 2 %, about 1.5 % to about 2.5 %, about 1.5 % to about3 %, about 1.5 % to about4 %, about 1.5 % to about 5 %, about 2 % to about 2.5 %, about 2 % to about 3 %, about 2 % to about 4 %, about 2 % to about 5 %, about 2.5 % to about 3 %, about 2.5 % to about 4 %, about 2.5 % to about 5 %, about 3 % to about 4 %, about 3 % to about 5 %, or about 4 % to about 5 % (w/w) of the composition.
  • the sterol is present in an amount of about 1 %, about 1.5 %, about 2 %, about 2.5 %, about 3 %, about 4 %, or about 5 %(w/w) of the composition. In some embodiments, the sterol is present in an amount of at least about 1 %, about 1.5 %, about 2 %, about 2.5 %, about 3 %, or about 4 % (w/w) of the composition. In some embodiments, the sterol is present in an amount of at most about 1.5 %, about 2 %, about 2.5 %, about 3 %, about 4 %, or about 5 % (w/w) of the composition.
  • the lipid vesicle compositions comprise one or more penetration enhancers.
  • Permeation enhancers act to increase the amount of penetration of an anionic polymer material, a peptide, or a combination thereof through one or more layers of skin when applied to the skin of an individual.
  • the penetration enhancer is included in the oil -in-water emulsion of the composition. In some embodiments, the penetration enhancer is included in the lipid bilayer of the composition.
  • the penetration enhancing agent comprising an ionic surfactant, a nonionic surfactant, or a combination thereof.
  • the penetration enhancing agent comprises a non-ionic surfactant or a combination of non-ionic surfactants.
  • the penetration enhancing agent is a single non-ionic surfactant.
  • the penetration enhancing agent is a combination of at least 2, 3, 4, or more non-ionic surfactants.
  • the penetration enhancing agent is a combination 2 non -ionic surfactants.
  • the penetration enhancing agent is a combination 3 non -ionic surfactants.
  • the non-ionic surfactant or combination of non-ionic surfactants is selected from polyethylene glycol ethers of fatty alcohols, sorbitan esters, polysorbates, and polyethylene glycol fatty acid esters and combinations thereof.
  • the combination of non-ionic surfactants is a combination of a polyethylene glycol ether of a fatty alcohol and a sorbitan ester. In some embodiments, the combination of non-ionic surfactants is a combination of a polyethylene glycol ethers of fatty alcohol and a polysorbate. In some embodiments, the combination of non-ionic surfactants is a combination of a polyethylene glycol ethers of fatty alcohol and a sorbitan ester. In some embodiments, the combination of non-ionic surfactants is a combination of a polyethylene glycol ethers of fatty alcohol and a polyethylene glycol fatty acid ester.
  • the combination of non-ionic surfactants is a combination of a polyethylene glycol ether of a fatty alcohol, a sorbitan ester, and a polysorbate. In some embodiments, the combination of non-ionic surfactants is a combination of a polyethylene glycol ether of a fatty alcohol, a sorbitan ester, and a polyethylene glycol fatty acid ester. In some embodiments, the combination of non-ionic surfactants is a combination of a polyethylene glycol ether of a fatty alcohol, a polysorbate, and a polyethylene glycol fatty acid ester.
  • the combination of non -ionic surfactants comprises a polyethylene glycol fatty acid ester and a sorbitan ester. In some embodiments, the combination of non-ionic surfactants comprises a polyethylene glycol fatty acid ester and a polysorbate. In some embodiments, the combination of non -ionic surfactants is a combination of a polyethylene glycol fatty acid ester, a polysorbate, and a sorbitan ester.
  • the non-ionic surfactant comprises a polyethylene glycol (PEG) ether of a fatty alcohol.
  • the PEG ether of the fatty alcohol comprises from about 2 to about 8 PEG groups and a C12-C22 fatty alcohol.
  • the polyethylene glycol ether of a fatty alcohol comprises diethylene glycol hexadecyl ether, 2-(2-octadecoxy ethoxy )ethanol, diethylene glycol monooleyl ether, polyoxyethylene (2) oleyl ether, polyoxyethylene (3) oleyl ether, or polyoxyethylene (5) oleyl ether, or any combination thereof.
  • the polyethylene glycol ether of a fatty alcohol comprises 2-(2-octadecoxyethoxy)ethanol.
  • the PEG ether of a fatty alcohol is super refined Brij® 02 or a derivative thereof.
  • the PEG ether of the fatty alcohol is present in an amount of from about 0.5 % to about 10 % (w/w) of the composition. In some embodiments, the PEG ether of the fatty alcohol is present in an amount of about 0.5 % to about 2.5 %.
  • the PEG ether of the fatty alcohol is present in an amount of about 0.5 % to about 0.8 %, about 0.5 % to about 1 %, about 0.5 % to about 1.2 %, about 0.5 % to about 1.5 %, about0.5 % to about2 %, about0.5 % to about2.5 %, about0.8 % to about 1 %, about 0.8 % to about 1.2 %, about 0.8 % to about 1.5 %, about 0.8 % to about 2 %, about 0.8 % to about 2.5 %, about 1 % to about 1.2 %, about 1 % to about 1.5 %, about 1 % to about 2 %, about 1 % to about 2.5 %, about 1.2 % to about 1.5 %, about 1.2 % to about2 %, about 1.2 % to about 2.5 %, about 1.5 % to about 2 %, about 1.5 % to about 2.5 %, or about 2 % to about 2.5 %.
  • the PEG ether of the fatty alcohol is present in an amount of about 0.5 %, about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, about 2 %, or about 2.5 %. In some embodiments, the PEG ether of the fatty alcohol is present in an amount of at least about 0.5 %, about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, or about 2 %. In some embodiments, the PEG ether of the fatty alcohol is present in an amount of at most about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, about 2 %, or about2.5 %.
  • the non-ionic surfactant comprises a fatty acid ester.
  • the non-ionic surfactant comprises PPG benzyl ether myristate, ethylhexyl stearate, isostearyl isostearate (e.g., Crodamol ISIS), myristyl myristate (e.g., Crodamol MM), glyceryl stearate (e.g., Crodamol GMS, Cithrol GMS).
  • the non ionic surfactant comprises a sorbitan ester.
  • the sorbitan ester is a fatty acid ester.
  • the sorbitan ester is a C12-C22 fatty acid ester. In some embodiments, the sorbitan ester comprises sorbitan monolaurate, sorbitan monopalm itate, sorbitan monostearate, sorbitan monooleate, sorbitan trioleate, sorbitan sesquioleate, or sorbitan isostearate, or any combinations thereof. In some embodiments, the sorbitan ester comprises sorbitan monolaurate. In some embodiments, the sorbitan ester comprises sorbitan monopalmitate. In some embodiments, the sorbitan ester comprises sorbitan monostearate.
  • the sorbitan ester comprises sorbitan monooleate. In some embodiments, the sorbitan ester comprises sorbitan trioleate. In some embodiments, the sorbitan ester comprises sorbitan sesquioleate. In some embodiments, the sorbitan ester comprises sorbitan isostearate.
  • the sorbitan ester is present in an amount of about 0. 1 % to about 2.5 % (w/w) of the composition. In some embodiments, the sorbitan ester is present in an amount of about 0.1 % to about 0.3 %, about 0.1 % to about 0.5 %, about 0.1 % to about 0.8 %, about 0.1 % to about 1 %, about 0.1 % to about 1.2 %, about 0.1 % to about 1.5 %, about 0.1 % to about 2 %, about 0.1 % to about 2.5 %, about 0.3% to about 0.5 %, about 0.3 % to about 0.8 %, about 0.3 % to about 1 %, about 0.3 % to about 1.2 %, about 0.3 % to about 1.5 %, about0.3 % to about2 %, about0.3 % to about2.5 %, about0.5 % to about0.8 %, about 0.5 % to about 1 %, about 0.5 % to about 1 %, about 0.5
  • the sorbitan ester is present in an amount of about 0.1%, about 0.3 %, about 0.5 %, about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, about 2 %, or about 2.5 %. In some embodiments, the sorbitan ester is present in an amount of at least about 0.1%, about 0.3 %, about 0.5 %, about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, or about2 %.
  • the sorbitan ester is present in an amount of at most about 0.1%, about 0.3%, about 0.5%, about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, about 2 %, or about 2.5 %.
  • the non-ionic surfactant comprises a polysorbate.
  • the polysorbate comprises polysorbate 20, polysorbate 21, polysorbate 40, polysorbate 60, polysorbate 80, polysorbate 85, or any combination thereof.
  • the polysorbate is polysorbate 80.
  • the polysorbate is polysorbate 20.
  • the polysorbate is present in an amount of about 0. 1 % to about
  • the polysorbate is present in an amount of about 0.1 % to about 0.2%, about 0.1 % to about 0.3 %, about 0.1 % to about 0.5 %, about 0.1 % to about 0.8 %, about 0.1 % to about 1 %, about 0.1 % to about 1.2 %, about 0.1 % to about 1.5 %, about 0.1 % to about 2 %, about 0.1 % to about 2.5 %, about 0.2 % to about 0.3 %, about 0.2 % to about 0.5 %, about 0.2 % to about 0.8 %, about 0.2 % to about 1 %, about 0.2 % to about 1.2 %, about 0.2 % to about 1.5 %, about 0.2 % to about 2 %, about 0.2 % to about 2.5 %, about 0.3 % to about 0.5 %, about 0.2 % to about 0.8 %, about 0.2 % to about 1 %, about 0.2 % to about 1.2 %, about 0.2
  • the polysorbate is present in an amount of about 0.1 %, about 0.2 %, about 0.3%, about 0.5 %, about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, about 2 %, or about 2.5 %.
  • the polysorbate is present in an amount of at least about 0.1 %, about 0.2 %, about 0.3%, about 0.5 %, about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, or about 2 %. In some embodiments, the polysorbate is present in an amount of at most about 0.2 %, about 0.3%, about 0.5%, about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, about 2 %, or about 2.5 %.
  • the non-ionic surfactant comprises a polyethylene glycol (PEG) fatty acid ester.
  • PEG polyethylene glycol
  • the PEG fatty acid ester is a PEG chain of about 2-8 subunits comprising C8-C22 fatty acids affixed to each terminal hydroxyl to form the fatty acid ester.
  • the PEG fatty acid ester comprises PEG-8 dilaurate, PEG-4 dilaurate, PEG-4 laurate, PEG-8 dioleate, PEG-8 distearate, PEG-8 distearate, PEG-7 glyceryl cocoate, and PEG-20 almond glycerides, or any combination thereof In some embodiments, the PEG fatty acid ester is PEG-4 dilaurate.
  • the PEG fatty acid ester is present in an amount of about 0.5 % to about 2.5 % (w/w) of the composition. In some embodiments, the PEG fatty acid ester is present in an amount of about 0.5 % to about 0.8 %, about 0.5 % to about 1 %, about 0.5 % to about 1.2 %, about0.5 % to about 1.5 %, about0.5 % to about2 %, about0.5 % to about2.5 %, about 0.8 % to about 1 %, about 0.8 % to about 1.2 %, about 0.8 % to about 1.5 %, about 0.8 % to about 2 %, about 0.8 % to about 2.5 %, about 1 % to about 1.2 %, about 1 % to about 1.5 %, about 1 % to about 2 %, about 1 % to about 2.5 %, about 1.2 % to about 1.5 %, about 1 % to about 2 %, about 1 % to about 2.5 %, about 1.2 % to about 1.5
  • the PEG fatty acid ester is present in an amount of about 0.5 %, about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, about 2 %, or about 2.5 %. In some embodiments, the PEG fatty ester is present in an amount of at least about 0.5 %, about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, or about 2 %. In some embodiments, the PEG fatty acid ester is present in an amount of at most about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, about2 %, or about2.5 %.
  • the non-ionic surfactant has a hydrophobic-lipophilic balance (HLB) of about 10 or less.
  • the non -ionic surfactant may be Cithrol GMS 40.
  • the composition comprises a plurality of non -ionic surfactants, each having an HLB of about 10 or less.
  • the non -ionic surfactant with an HLB of 10 or less is selected from the Table 1, or any combination thereof.
  • the non-ionic surfactant or combination of non-ionic surfactants are present in an amount of about 0.5 % to about 10 % (w/w) of the composition.
  • the non-ionic surfactant or combination of non-ionic surfactants are present in an amount of about 0.5 % to about 1 %, about 0.5 % to about 1.5 %, about 0.5 % to about 2 %, about 0.5 % to about 3 %, about 0.5 % to about 4 %, about 0.5 % to about 5 %, about 0.5 % to about 6 %, about 0.5 % to about 7 %, about 0.5 % to about 8 %, about 0.5 % to about 10 %, about 1 % to about 1.5 %, about 1 % to about 2 %, about 1 % to about 3 %, about 1 % to about 4 %, about 1 % to about 5 %, about 1 % to about 6 %, about 1 % to about 7 %, about 1 % to about 8 %, about 1 % to about 10 %, about 1.5 % to about 2 %, about 1.5 % to about 3 %, about 1.5 % to about % to about
  • the non -ionic surfactant or combination of non-ionic surfactants are present in an amount of about 0.5 %, about 1 %, about 1.5 %, about 2 %, about 3 %, about 4 %, about 5 %, about 6 %, about 7 %, about 8 %, or about 10 %. In some embodiments, the non -ionic surfactant or combination of non-ionic surfactants are present in an amount of at least about 0.5 %, about 1 %, about 1.5 %, about 2 %, about 3 %, about 4 %, about 5 %, about 6 %, about 7 %, or about 8 %.
  • the non-ionic surfactant or combination of non-ionic surfactants are present in an amount of at most about 1 %, about 1.5 %, about 2 %, about 3 %, about 4 %, about 5 %, about 6 %, about 7 %, about 8 %, or about 10 %.
  • the composition comprises a non-ionic surfactant in the oil-in water emulsion, the lipid bilayer, or both. In some embodiments, the composition comprises a non-ionic surfactant in the oil -in-water emulsion. In some embodiments, the composition comprises a non-ionic surfactant in the lipid bilayer. In some embodiments, the composition comprises a non-ionic surfactant in the oil-in-water emulsion and the lipid bilayer, wherein the composition comprises two or more different non-ionic surfactants.
  • the penetration enhancing agent comprises a salicylate ester or a nicotinate ester.
  • the ester is a C1-C6 alkyl ester or a benzyl ester.
  • the penetration enhancing agent comprises methyl salicylate or benzyl nicotinate.
  • the penetration enhancing agent is a nicotinate ester present in an amount of up to about 0.1 %, 0.5%, 1%, 2%, or 3% (w/w) of the composition.
  • the nicotinate ester is present in an amount of from about 0.1% to about 3%, about 0.1% to about 2%, or about 0.1% to about 1%.
  • the penetration enhancing agent comprises a fatty acid acylated amino acid.
  • the fatty acid acylated amino acid is lysine.
  • the lysine is mono-acylated with a fatty acid.
  • the penetration enhancing agent is monoloauryl lysine.
  • the lysine is di- acylated.
  • the penetration enhancing agent is dipalmitoyllysine.
  • the fatty acylated amino acid is present in an amount of about 1%, about 2%, about 3%, about 4%, or about 5% (w/w) of the composition.
  • the fatty acylated amino acid is present in an amount of at least about 1%, about 2%, about 3%, about 4%, or about 5% (w/w) of the composition. In some embodiments, the fatty acylated amino acid is present in an amount of up to about 1%, up to about 2%, up to about 3%, up to about 4%, or up to about 5% (w/w) of the composition.
  • the fatty acylated amino acid is present in an amount of from about 0.1% to about 5%, from about 0.1% to about 4%, from about 0.1% to about 3%, from about 0.1% to about 2%, from about 0.5% to about 5%, from about0.5% to about 4%, from about0.5% to about3%, from about 0.5% to about 2%, from about 1% to about 5%, from about 1% to about 4%, from about 1% to about 3%, from about 1% to about2%, or from about 1.5% to about2.5%.
  • the non-ionic surfactant has a hydrophobic-lipophilic balance (HLB) of about 10 or less.
  • the composition comprises a plurality of non-ionic surfactants, each having an HLB of about 10 or less.
  • the non-ionic surfactant with an HLB of 10 or less is selected from the Table A below, or any combination thereof.
  • the non-ionic surfactant has a hydrophobic-lipophilic balance (HLB) of about 10 or more.
  • the composition comprises a plurality of non-ionic surfactants, each having an HLB of about 10 or more.
  • the non-ionic surfactant or combination of non-ionic surfactants are present in an amount of about 0.5 % to about 10 % (w/w) of the composition. In some embodiments, the non -ionic surfactant or combination of non-ionic surfactants are present in an amount of about 0.5 % to about 1 %, about 0.5 % to about 1.5 %, about 0.5 % to about 2 %, about 0.5 % to about 3 %, about 0.5 % to about 4 %, about 0.5 % to about 5 %, about 0.5 % to about 6 %, about 0.5 % to about 7 %, about 0.5 % to about 8 %, about 0.5 % to about 10 %, about 1 % to about 1.5 %, about 1 % to about 2 %, about 1 % to about 3 %, about 1 % to about 4 %, about 1 % to about 5 %, about 1 % to about 6 %, about 1 % to about 10 %, about
  • the non -ionic surfactant or combination of non -ionic surfactants are present in an amount of about 0.5 %, about 1 %, about 1.5 %, about 2 %, about 3 %, about 4 %, about 5 %, about 6 %, about 7 %, about 8 %, or about 10 %. In some embodiments, the non -ionic surfactant or combination of non-ionic surfactants are present in an amount of at least about 0.5 %, about 1 %, about 1.5 %, about 2 %, about 3 %, about 4 %, about 5 %, about 6 %, about 7 %, or about 8 %.
  • the non-ionic surfactant or combination of non-ionic surfactants are present in an amount of at most about 1 %, about 1.5 %, about 2 %, about 3 %, about 4 %, about 5 %, about 6 %, about 7 %, about 8 %, or about 10 %.
  • the composition comprises a non-ionic surfactant in the oil-in water emulsion, the lipid bilayer, or both. In some embodiments, the composition comprises a non-ionic surfactant in the oil-in-water emulsion. In some embodiments, the composition comprises a non-ionic surfactant in the lipid bilayer. In some embodiments, the composition comprises a non-ionic surfactant in the oil-in-water emulsion and the lipid bilayer, wherein the composition comprises two or more different non -ionic surfactants.
  • the penetration enhancing agent comprises a salicylate ester or a nicotinate ester.
  • the ester is a Ci-C 6 alkyl ester or a benzyl ester.
  • the penetration enhancing agent comprises methyl salicylate or benzyl nicotinate.
  • the penetration enhancing agent is a nicotinate ester present in an amount of up to about 0.1 %, 0.5%, 1%, 2%, or 3% (w/w) of the composition.
  • the nicotinate ester is present in an amount of from about 0.1% to about 3%, about 0.1% to about 2%, or about 0.1% to about 1%.
  • the composition comprises an ionic surfactant.
  • the ionic surfactant is a cationic surfactant.
  • the cationic surfactant is a mono-cationic surfactant, a di-cationic surfactant, or a poly -cationic surfactant.
  • the mono-cationic surfactant is used in the composition to form a submicron emulsion prior to formation of a final lipid vesicle composition provided herein (e.g., before the lipid forming vesicles are added).
  • the mono-cationic surfactant is net-mono-cationic (e.g., a phosphate salt comprising two side chains each with a single cationic functionality, which is partially neutralized by a phosphate anion).
  • the mono-cationic surfactant is a fatty-amide derived propylene glycol-diammonium phosphate ester.
  • Fatty-amide derived propylene glycol- diammonium phosphate esters are phospholipids which comprise at least one propylene glycol phosphoester linked to a quaternary ammonium group, which is in turn linked with a fatty acid amide.
  • a fatty -amide derived propylene glycol- diammonium phosphate ester is linoleamidopropyl PG-dimonium chloride phosphate. Similar compounds with different fatty acid amide groups attached are also known.
  • the fatty-amide derived propylene glycol-diammoniom phosphate ester has the structure: wherein n is an integer from 1 to 3, m is an integer from 0 to 2, wherein the sum of m and n is 3; X is a cation selected from a proton, sodium, potassium, magnesium, and calcium; and R is an acyl group of a C 8 -C 30 fatty acid.
  • the fatty acid is a C 12 -C 24 fatty acid. In some embodiments, the fatty acid is an unsaturated fatty acid. In some embodiments, the fatty acid is linoleic acid. In some embodiments, the mono-cationic penetration enhancing agent is linoleamidopropyl PG- dimonium chloride phosphate (e.g., ArlasilkTM PTM, ArlasilkTMEFA).
  • the fatty amide derived propylene glycol -diammonium phosphate ester is present in an amount of about 1 % to about 10 % (w/w) of the composition. In some embodiments, the fatty amide derived propylene glycol -diammonium phosphate ester is present in an amount of about 1 % to about 2 %, about 1 % to about 3 %, about 1 % to about 4 %, about 1 % to about 5 %, about 1 % to about 6 %, about 1 % to about 7 %, about 1 % to about 8 %, about 1 % to about 9 %, about 1 % to about 10 %, about 2 % to about 3 %, about 2 % to about 4 %, about 2 % to about 5 %, about 2 % to about 6 %, about 2 % to about 7 %, about 2 % to about 8 %, about 2 % to about 9 %, about 2 % to about 10 %, about
  • the fatty amide derived propylene glycol-diammonium phosphate ester is present in an amount of about 1 %, about 1.5%, about2 %, about2.5%, about3 %, about3.5%, about4 %, about4.5%, about 5 %, about 5.5%, about 6 %, about 6.5%, about 7 %, about 7.5%, about 8 %, about 8.5%, about 9 %, about 9.5%, or about 10 % (w/w) of the composition.
  • the fatty amide derived propylene glycol-diammonium phosphate ester is present in an amount of at least about 1 %, about 1.5%, about2 %, about2.5%, about3 %, about 3.5%, about 4 %, about 4.5%, about 5 %, about 5.5%, about 6 %, about 6.5%, about 7 %, about 7.5%, about 8 %, about 8.5%, about 9 %, or about 9.5%,.
  • the fatty amide derived propylene glycol -diammonium phosphate ester is present in an amount of at most about 1.5%, about 2 %, about 2.5%, about 3 %, about3.5%, about4 %, about4.5%, about 5 %, about 5.5%, about6 %, about6.5%, about7 %, about7.5%, about8 %, about8.5%, about9 %, about9.5%, or about 10 %.
  • the cationic surfactant is a di-cationic penetration enhancing agent.
  • the di-cationic surfactant is a gemini surfactant.
  • a gemini surfactant is a surfactant comprising two quaternary amines represented by the formula , wherein each of A and C is independently an optionally substituted C 6 -C 2 4 alkyl group, each R is independently optionally substituted Ci-C 6 alkyl, andB is an optionally substituted C 2 -Ci 0 alkylene chain.
  • the each of A and C is a C 6 -C 2 saturated or unsaturated hydrocarbon.
  • each of A and C is a C 6 -C 24 saturated hydrocarbon.
  • each R is methyl.
  • B is a saturated C 2 -Ci 0 alkylene chain.
  • gemini surfactants follow the nomenclature X-Y-Z, wherein each of X, Y, and Z is an integer representing the number of carbon atoms of each substituent, and Y is the spacer between the two quaternary amines.
  • a 12-3-12 gemini surfactant has the formula CH 3 (CH 2 )11- P4 + (CH 3 ) 2 ]-(CH 2 ) 3 -[N + (CH 3 ) 2 ]-(CH 2 ) II CH 3.
  • the gemini surfactant is a 10-2-10, 12-2-12, 14-2-14, 10-3-10, 12-3-12, 14-3-14, 10-4-10, 12-4-12, or 14-4-14 gemini surfactant.
  • the gemini surfactant is a 12-3-12 gemini surfactant.
  • the gemini surfactant is present in an amount of about 0.1 % to about 1.5 % (w/w) of the composition. In some embodiments, the gemini surfactant is present in an amount of about 0.1 % to about 0.2 %, about 0.1 % to about 0.3 %, about 0.1 % to about 0.5 %, about 0.1 % to about 0.7 %, about 0.1 % to about 0.9 %, about 0.1 % to about 1 %, about 0.1 % to about 1.2 %, about 0.1 % to about 1.5 %, about 0.2 % to about 0.3 %, about 0.2 % to about 0.5 %, about 0.2 % to about 0.7 %, about 0.2 % to about 0.9 %, about 0.2 % to about 1 %, about 0.2 % to about 1.2 %, about 0.2 % to about 1.5 %, about 0.3 % to about 0.5 %, about 0.2 % to about 0.7 %, about 0.2 % to
  • the gemini surfactant is present in an amount of about 0.1 %, about 0.2 %, about 0.3 %, about 0.5 %, about 0.7 %, about 0.9 %, about 1 %, about 1.2 %, or about 1.5 %. In some embodiments, the gemini surfactant is present in an amount of at least about 0.1 %, about 0.2 %, about 0.3 %, about 0.5 %, about 0.7 %, about 0.9 %, about 1 %, or about 1.2 %.
  • the gemini surfactant is present in an amount of at most about 0.2 %, about 0.3 %, about 0.5 %, about 0.7 %, about 0.9 %, about 1 %, about 1.2 %, or about 1.5 %.
  • the cationic surfactant comprises a poly cationic group.
  • the poly cationic group is a polymer wherein each monomer of the polymer comprises a charged group (e.g., an amino group).
  • the poly cationic group is polylysine.
  • the polycationic group is polyarginine.
  • the cationic surfactant may comprise an amino acid and a fatty acid.
  • the cationic surfactant may be derived from an amino acid and a fatty acid.
  • the amino acid may comprise a cationic amino acid, such as lysine, arginine, or histidine.
  • the fatty acid may comprise a undecyloyl, lauroyl, tridecyloyl, myristoyl, palmitoyl, or stearoyl group
  • the cationic surfactant may comprise lauroyl lysine or lauroyl arginine.
  • the polylysine has a molecular weight of from about 1 kDa to about 10 kDa, from about 1 kDa to about 5 kDa, or from about 3 kDa to about 5 kDa. In some embodiments, the polylysine is present in an amount of from about 0.01% to about 1%, from about 0.01% to about0.5%, from about 0.01% to about 0.2%, from about0.05% to about 1%, from about 0.05% to about 0.5%, or from about 0.05% to about 0.2% (w/w) of the composition.
  • the vesicle composition comprises additional components. In some embodiments, these additional components improve one or more properties of the vesicles without dramatically altering the delivery of an active ingredient.
  • the active ingredient comprises a peptide antagonist of a melanocortin 1 receptor. In some embodiments, the peptide antagonist is present in the composition in an amount of about 0.1 % to about 1.5 % (w/w).
  • the peptide antagonist is present in an amount of about 0.1 % to about 0.2 %, about 0.1 % to about 0.3 %, about 0.1 % to about 0.5 %, about 0.1 % to about 0.7 %, about 0.1 % to about 0.9 %, about 0.1 % to about 1 %, about 0.1 % to about 1.2 %, about 0.1 % to about 1.5 %, about 0.2 % to about 0.3 %, about 0.2 % to about 0.5 %, about 0.2 % to about 0.7 %, about 0.2 % to about 0.9 %, about 0.2 % to about 1 %, about 0.2 % to about 1.2 %, about 0.2 % to about 1.5 %, about 0.3 % to about 0.5 %, about 0.3 % to about 0.7 %, about 0.3 % to about 0.9 %, about 0.3 % to about 1 %, about 0.2 % to about 1.2 %, about 0.2 % to about 1.5
  • the peptide antagonist is present in an amount of about 0.1 %, about 0.2 %, about 0.3 %, about 0.5 %, about 0.7 %, about 0.9 %, about 1 %, about 1.2 %, or about 1.5 %. In some embodiments, the peptide antagonist is present in an amount of at least about 0.1 %, about 0.2 %, about 0.3 %, about 0.5 %, about 0.7 %, about 0.9 %, about 1 %, or about 1.2 %.
  • the peptide antagonist is present in an amount of at most about 0.2 %, about 0.3 %, about 0.5 %, about 0.7 %, about 0.9 %, about 1 %, about 1.2 %, or about 1.5 %.
  • the vesicle composition further comprises one or more viscosity enhancing agents.
  • the viscosity enhancing agents thicken the composition for increased stability and/or feel to a user of the vesicle composition.
  • the viscosity enhancing agents also act as surfactants.
  • the viscosity enhancing agent comprises one or more of a fatty alcohol, a wax, a fatty ester of glycerol, or any combination thereof.
  • the fatty alcohol is a C 8 -C 2 o fatty alcohol.
  • the fatty alcohol is cetyl alcohol.
  • the cetyl alcohol is Crodacol C95.
  • the wax is a naturally occurring or synthetic wax.
  • the wax is beeswax.
  • the wax is synthetic beeswax.
  • the synethetic beeswax is syncrowaxTMBB4.
  • the synthetic beeswax is non-animal derived beeswax.
  • the non-animal derived beeswax is syncrowaxTM SB1.
  • the wax comprises jojoba wax, candelilla wax, carnaubawax, or vegetable oil.
  • the fatty ester of glycerol is a monoester.
  • the monoester is an ester of a C8-C24 fatty acid.
  • the fatty ester of glycerol is glycerol monostearate.
  • the viscosity enhancing agents are present in an amount of from about 0.5% to about 10% (w/w) of the composition. In some embodiments, the viscosity enhancing agents are present in an amount of from about 0.5% to about 5%, about 0.5 % to about 5%, about 0.5 % to about 4%, about 0.5 % to about 3%, or from about 0.5% to about 2% (w/w) of the composition. In some embodiments, the viscosity enhancing agents comprise a fatty alcohol in an amount of up to about 2 %, a wax in an amount of up to about 2%, and a fatty ester of glycerol in an amount of up to about 5 %.
  • the fatty alcohol is present in an amount of from about 0.1 to about 1.5%. In some embodiments, the fatty alcohol is present in an amount of about 0.4%. In some embodiments, the wax is present in an amount of from about 0.1% to about 1%. In some embodiments, the wax is present in an amount of about 0.2 %. In some embodiments, two or more waxes are each present in an amount of about 0.2 %. In some embodiments, the fatty ester of glycerol is present in an amount of from about 0.5 % to about 2 %.
  • the vesicle composition further comprises one or more of a thickener, a preservative, a moisturizer, an emollient, a humectant, or any combination thereof.
  • the vesicle composition further comprises a thickener.
  • the vesicle composition further comprises a preservative.
  • the vesicle composition further comprises a moisturizer.
  • the vesicle composition further comprises an emollient.
  • the emollient may be derived from an amino acid and a fatty acid.
  • the amino acid may comprise lysine, arginine, histidine, glutamate, aspartate, or sarcosinate.
  • the emollient may comprise lauroyl lysine, lauroyl arginine, isopropyl lauroyl sarcosinate, phytosteryl lauroyl glutamate, behenyl lauroyl glutamate, octyldodecyl lauroyl glutamate, or cholesteryl lauroyl glutamate.
  • the vesicle composition further comprises a humectant.
  • the vesicle composition further comprises an antimicrobial.
  • the antimicrobial is a paraben ester.
  • the antimicrobial is methylparaben or propylparaben, or a combination thereof.
  • the antimicrobial is present in an amount of up to about 1%, up to about 0.9%, up to about 0.8%, up to about 0.7%, up to about 0.6%, up to about 0.5%, up to about 0.4%, up to about 0.3%, up to about 0.2% (w/w) of the composition.
  • the vesicle composition further comprises a thickener.
  • the thickener is an inert polymer material.
  • the thickener is a siloxane polymer.
  • the thickener poly dimethyl siloxane (PDMS).
  • the PDMS is present in an amount of up to about 5%, up to about 4%, up to about 3%, up to about 2%, or up to about 1%. In some embodiments, the PDMS is present in an amount of from about 0.1% to about 2% (w/w) of the composition.
  • the vesicle composition further comprises a humectant.
  • the composition comprises glycerol.
  • the glycerol is present in an amount of from about 0.5 % to about 25 %, about 0.5 % to about 20%, about 0.5 % to about 15 %, or about 0.5 % to about 10 % (w/w) of the composition.
  • the glycerol is present in an amount of about 1 % to about 10 %.
  • the glycerol is present in an amount of about 1 % to about 2 %, about 1 % to about 4 %, about 1 % to about 6 %, about 1 % to about 8 %, about 1 % to about 10 %, about 2 % to about 4 %, about 2 % to about 6 %, about 2 % to about 8 %, about 2 % to about 10 %, about 4 % to about 6 %, about 4 % to about 8 %, about 4 % to about 10 %, about 6 % to about 8 %, about 6 % to about 10 %, or about 8 % to about 10 %.
  • the glycerol is present in an amount of about 1 %, about 2 %, about 4 %, about 6 %, about 8 %, or about 10 %. In some embodiments, the glycerol is present in an amount of at least about 1 %, about 2 %, about 4 %, about 6 %, or about 8 %. In some embodiments, the glycerol is present in an amount of at most about 2 %, about 4 %, about 6 %, about 8 %, or about 10 %. [0298] In some embodiments, the vesicle composition further comprises a preservative. In some embodiments, the preservative is a paraben ester.
  • the preservative is methylparaben or propylparaben, or a combination thereof. In some embodiments, the preservative is present in an amount of up to about 1%, up to about 0.9%, up to about 0.8%, up to about 0.7%, up to about 0.6%, up to about 0.5%, up to about 0.4%, up to about 0.3%, up to about 0.2% (w/w) of the composition. In some embodiments, the preservative is a cosmetic preservative, such as Euxyl® PE 9010 or Spectrastat®. In some embodiments, the preservative comprises a phenoxyethanol/ethylhexylglycerin mixture.
  • the preservative comprises a blend of caprylhydroxamic acid, caprylyl glycol, and glycerin. In some embodiments, the preservative is present in an amount of up to about 2%, up to about 1.5 %, or up to about 1% (w/w) of the composition. In some embodiments, the preservative is present in an amount of from about 0.1% to about 2%, from about 0.1% to about 1.5%, or from about 0.1% to about 1%. In some embodiments, the preservative is present in an amount of about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, or 1.5%..
  • an additional component in the lipid vesicle composition may comprise a molecule that can directly or indirectly reduce melanogenesis.
  • the molecule may directly or indirectly inhibit tyrosinase.
  • the additional component is an antioxidant.
  • the antioxidant may comprise vitamin C or a derivative thereof.
  • the antioxidant may comprise ascorbic acid, ascorbyl palmitate, sodium ascorbyl phosphate, magnesium ascorbyl phosphate, retinyl ascorbate, aminopropyl ascorbyl phosphate (K3 Vita-C), tetrahexyldecyl ascorbate, ascorbyl methylsilanol pectinate (ascorbosilane SP), or a combination thereof.
  • the antioxidant such as a vitamin C or derivative thereof, may also aid in collagen synthesis.
  • the additional component may comprise resorcinol (e.g., 4-butyl resorcinol).
  • 4-butyl resorcinol may be used to reduce hyperpigmentation in a layer of the skin, such as the epidermis.
  • resorcinol is present in the composition in an amount of from about 0.005 % to about 0.1%.
  • resorcinol is present in the composition in an amount of from about 0.005 % to about 0.01 %, about 0.005 % to about 0.02 %, about 0.005 % to about 0.05 %, about 0.005 % to about 0.07 %, about0.005 % to about 0.1 %, about 0.01 % to about0.02 %, about 0.01 % to about 0.05 %, about 0.01 % to about 0.07 %, about 0.01 % to about 0.1 %, about 0.02 % to about 0.05 %, about 0.02 % to about 0.07 %, about 0.02 % to about 0.1 %, about 0.05 % to about 0.07 %, about 0.05 % to about 0.1 %, or about 0.07 % to about 0.1 %.
  • resorcinol is present in the composition in an amount of about 0.005 %, about 0.01 %, about 0.02 %, about 0.05 %, about 0.07 %, or about 0.1 %. In some embodiments, resorcinol is present in the composition in an amount of at least about 0.005 %, about 0.01 %, about 0.02 %, about 0.05 %, or about 0.07 %. In some embodiments, resorcinol is present in the composition in an amount of at most about 0.01 %, about 0.02 %, about 0.05 %, about 0.07 %, or about 0.1 %.
  • an additional component may comprise one or more vitamins or derivatives thereof.
  • the one or more vitamins or derivatives thereof may provide one or more desired effects (e.g., smoothing, toning, moisturizing, brightening, reducing acne, eczema, wrinkles, etc.) when applied to the skin or epidermis.
  • the additional component may comprise vitamin B, vitamin C (ascorbic acid), vitamin A (retinol), vitamin E, vitamin D, vitamin F, vitamin K or any derivative thereof.
  • the additional component may comprise vitamin B3 (niacin).
  • a vitamin B3 derivative may comprise niacinamide.
  • the additional component may comprise provitamin B5 (panthenol).
  • the one or more vitamins or derivatives thereof are present in the composition in an about from about 0.1 % to about 2.5 % (w/w). In some embodiments, the one or more vitamins or derivatives thereof is present in an amount of about 0.
  • the one or more vitamins or derivatives thereof is present in an amount of about 0.1 %, about 0.2 %, about 0.3%, about 0.5 %, about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, about2 %, or about2.5 %. In some embodiments, the one or more vitamins or derivatives thereof is present in an amount of at least about 0.1 %, about 0.2 %, about 0.3%, about 0.5 %, about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, or about 2 %.
  • the one or more vitamins or derivatives thereof is present in an amount of at most about 0.2 %, about 0.3%, about 0.5%, about 0.8 %, about 1 %, about 1.2 %, about 1.5 %, about 2 %, or about 2.5 %.
  • the lipid vesicle composition provided herein comprise an anionic polymer material.
  • the anionic polymer material is desirably one which is compatible with delivery beneath the surface of the skin of a subject.
  • the anionic polymer material is one which acts as a volumizer or filler after delivery beneath the surface of the skin.
  • the anionic polymer material acts as a support for another layer of skin (e.g., the epidermis) in order to correct depressions of the skin or restore facial volume.
  • the anionic polymer material comprises an anionic polysaccharide.
  • the anionic polysaccharide is non-sulfatedglycosaminoglycan.
  • the anionic polymeric material is a naturally occurring substance.
  • the anionic polymeric material naturally occurs in a human.
  • the anionic polymer material naturally occurs in connective or epithelial tissue in a human.
  • the anionic polymeric material is hyaluronic acid, or a pharmaceutically acceptable salt thereof.
  • the anionic polymer material may not be crosslinked in the lipid vesicle composition as described herein.
  • the hyaluronic acid is a pharmaceutically acceptable salt of hyaluronic acid.
  • the salt is the sodium salt, the potassium salt, the magnesium salt, or any combination thereof.
  • the salt is the sodium salt.
  • the anionic polymer material has a molecular weight of from about 5 kDato about 500 kDa. In some embodiments, the molecular weight is the weight average molecular weight. In some embodiments, the anionic polymeric material has a molecular weight of about 5 kDa to about 500 kDa.
  • the anionic polymeric material has a molecular weight of about 5 kDato about 10 kDa, about 5 kDato about 20 kDa, about 5 kDato about 50 kDa, about 5 kDa to about 100 kDa, about 5 kDa to about 200 kDa, about 5 kDa to about 250 kDa, about 5 kDato about 300 kDa, about 5 kDato about 400 kDa, about 5 kDato about500 kDa, about 10 kDato about20 kDa, about 10 kDato about 50 kDa, about lOkDato about 100 kDa, about 10 kDato about200kDa, about 10 kDato about250kDa, about lOkDato about 300 kDa, about 10 kDato about 400 kDa, about 10 kDato about 500 kD
  • the anionic polymeric material has a molecular weight of about 5 kDa, about 10 kDa, about 20 kDa, about 50 kDa, about 100 kDa, about 200 kDa, about 250 kDa, about 300 kDa, about 400 kDa, or about 500 kDa. In some embodiments, the anionic polymeric material has a molecular weight of at least about 5 kDa, about 10 kDa, about 20 kDa, about 50 kDa, about 100 kDa, about 200 kDa, about 250 kDa, about 300 kDa, or about 400 kDa.
  • the anionic polymeric material has a molecular weight of at most about 10 kDa, about 20 kDa, about 50 kDa, about 100 kDa, about200 kDa, about250 kDa, about300 kDa, about400 kDa, or about 500 kDa.
  • the lipid vesicle composition comprises a first and a second anionic polymer material. In some embodiments, the lipid vesicle composition further comprises a third anionic polymer material. [0306] In some embodiments, the first and the second anionic polymer material are the same type. In some embodiments, each of the first and the second anionic polymer material is an anionic polysaccharide. In some embodiments, each of the first and the second anionic polymer is hyaluronic acid.
  • each anionic polymer material has a different molecular weight.
  • the first anionic polymer material has a molecular weight of up to about 75 kDa and the second anionic polymer material has a molecule weight of greater than about 75 kDa.
  • the first anionic polymer material has a molecular weight of up to about 75 kDa and the second anionic polymer material has a molecular weight of greater than about 75 kDa.
  • each component may be included in a different amount.
  • the first and second anionic polymer material are present in about the same amount.
  • the ratio of the fist and the second anionic material is about 10:1, 9:1. 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 3:2, 2:1, or about 1 :1.
  • each of the anionic polymer materials can be of the same type (e.g., three different molecular weights of hyaluronic acid).
  • the composition comprises a first, second, and a third anionic polymer material, wherein the first anionic polymer material has a molecular weight of from about 5 kDa to about 20kDa, the second anionic polymer has a molecular weight of from about 20 kDa to about 75 kDa, and the third anionic polymer material has a molecular weight of greater than about 75 kDa.
  • each of the three anionic polymer materials is present in about the same amount.
  • the anionic polymer material is present in an amount of from about 0.01 mg/mLto about 10 mg/mL. In some embodiments, the anionic polymer material is present in an amount of about 0.01 mg/mL to about 0.05 mg/mL, about 0.01 mg/mLto about 0.1 mg/mL, aboutO.Ol mg/mLto about 0.5 mg/mL, aboutO.Ol mg/mLto about 1 mg/mL, about 0.01 mg/mL to about 1.25 mg/mL, aboutO.Ol mg/mL to about 1.5 mg/mL, aboutO.Ol mg/mLto about 1.75 mg/mL, aboutO.Ol mg/mLto about2 mg/mL, aboutO.Ol mg/mLto about5 mg/mL, aboutO.Ol mg/mLto about 10 mg/mL, about 0.05 mg/mLto about 0.1 mg/mL, about 0.05 mg/mL to about 0.5 mg/mL,
  • the anionic polymer material is present in an amount of about 0.01 mg/mL, about 0.05 mg/mL, about 0.1 mg/mL, about 0.5 mg/mL, about 1 mg/mL, about 1.25 mg/mL, about 1.5 mg/mL, about 1.75 mg/mL, about 2 mg/mL, about 5 mg/mL, or about 10 mg/mL.
  • the anionic polymer material is present in an amount of at least about 0.01 mg/mL, about 0.05 mg/mL, about 0.1 mg/mL, about 0.5 mg/mL, about 1 mg/mL, about 1.25 mg/mL, about 1.5 mg/mL, about 1.75 mg/mL, about 2 mg/mL, or about 5 mg/mL. In some embodiments, the anionic polymer material is present in an amount of at most about 0.05 mg/mL, about 0.1 mg/mL, about 0.5 mg/mL, about 1 mg/mL, about 1.25 mg/mL, about 1.5 mg/mL, about 1.75 mg/mL, about2 mg/mL, about5 mg/mL, or about 10 mg/mL.
  • the additional components comprise purified water.
  • purified water is present in an amount of about 50% to 80% (w/w).
  • purified water is present in an amount of about 50 % to about55 %, about50 % to about 60 %, about 50 % to about 65 %, about 50 % to about 70 %, about 50 % to about 75 %, about 50 % to about 80 %, about 55 % to about 60 %, about 55 % to about 65 %, about 55 % to about 70 %, about 55 % to about 75 %, about 55 % to about 80 %, about 60 % to about 65 %, about 60 % to about 70 %, about 60 % to about 75 %, about 60 % to about 80 %, about 65 % to about 70 %, about 65 % to about 75 %, about 65 % to about 80 %, about 70 % to about 75 %, about 70 % to about 80 %, about 70 % to about 75 %, about 70 %
  • purified water is presentin an amount of about 50 %, about 55 %, about 60 %, about 65 %, about 70 %, about 75 %, or about 80 %. In some embodiments, purified water is present in an amount of at least about 50 %, about 55 %, about 60 %, about 65 %, about 70 %, or about 75 %. In some embodiments, purified water is present in an amount of atmostabout55 %, about 60 %, about65 %, about70 %, about 75 %, or about 80 %.
  • compositions for the delivery of peptide antagonists are provided below.
  • the delivery of the peptide antagonist comprises a method of preparing a lipid vesicle composition, the method comprising: a) preparing an oil -in-water emulsion comprising the peptide antagonist of a melanocortin 1 receptor, by mixing oil components of the oil-in-water emulsion with aqueous components of the oil -in-water emulsion; b) solubilizing vesicle forming lipids in an acceptable solvent other than water; c) adding the oil-in-water emulsion to the solubilized vesicle forming lipids; and d) mixing the oil-in-water emulsion and the solubilized vesicle forming lipids under mixing conditions effective to form the lipid vesicles comprising a lipid bilayer comprising vesicle forming lipids, and an oil-in-water emulsion entrapped in the lipid vesicles.
  • the aqueous components comprise the peptide antagonist of a melanocortin 1 receptor.
  • the oil components and/or the aqueous components of the oil-in-water emulsion comprises the one or more surfactants.
  • the embodiments below may additional comprise any of the other ingredients or components provided herein.
  • Peptide Composition 1 In one aspect, provided herein, is a lipid vesicle composition comprising:
  • lipid vesicles each comprising a lipid bilayer comprising vesicle forming lipids, wherein the vesicle forming lipids are present in an amount of from about 5% to about 20%;
  • a peptide antagonist of a melanocortin 1 receptor in an amount of from about 0.1 mg/mL to about 50 mg/mL entrapped in the lipid bilayer and/or the oil-in-water emulsion, wherein the composition further comprises: a fatty amide derived propylene glycol-diammonium phosphate ester in an amount of from about 1% to about 10%; and a non-ionic surfactant in an amount of from about 0.1% to about 3%.
  • the oil component is present in an amount of from about 2.5% to about 20 %.
  • the lipid vesicle composition comprises the peptide antagonist in an amount of about 0.1 mg/mL to about 0.5 mg/mL, about 0.1 mg/mL to about 1 mg/mL, about 0.1 mg/mL to about 2 mg/mL, about 0.1 mg/mL to about 3 mg/mL, about 0.1 mg/mL to about 4 mg/mL, about 0.1 mg/mL to about 5 mg/mL, about 0.1 mg/mL to about 10 mg/mL, about 0.1 mg/mL to about20 mg/mL, about 0.1 mg/mL to about50 mg/mL.
  • the lipid vesicle composition comprises the peptides antagonist in an amount of about 0.1 mg/mL, about 0.5 mg/mL, about 1 mg/mL, about2 mg/mL, about 3 mg/mL, about 4 mg/mL, about 5 mg/mL, about 10 mg/mL, about 20 mg/mL, or about 50 mg/mL. In some embodiments, the lipid vesicle composition comprises the peptides antagonist in an amount of about 1 mg/mL, about 2 mg/mL, about 3 mg/mL, about 4 mg/mL, or about 5 mg/mL.
  • the composition further comprises a fatty acylated amino acid in an amount of from about 0.5 % to about 3%.
  • the fatty acylated amino acid is monoloauryl lysine.
  • the lipid vesicle composition further comprises viscosity enhancing agents in an amount of from about 0.5% to about 5%.
  • the viscosity enhancing agents comprise one or more of a fatty alcohol, a wax, a fatty ester of glycerol, or any combination thereof.
  • the non-ionic surfactant comprises a PEG ether of a fatty alcohol.
  • the lipid vesicle composition further comprises an anionic polymer material in an amount of from about 0.01 mg/mL to about 10 mg/mL entrapped in the lipid bilayer, the oil-in-water emulsion, or a combination thereof.
  • the lipid vesicle composition comprises the anionic polymer material in an amount of about 0.01 mg/mL to about0.05 mg/mL, about 0.01 mg/mL to about 0.1 mg/mL, about 0.01 mg/mL to about 0.5 mg/mL, about 0.01 mg/mL to about 1 mg/mL, about 0.01 mg/mL to about 1.25 mg/mL, about 0.01 mg/mL to about 1.5 mg/mL, about 0.01 mg/mL to about 1.75 mg/mL, about 0.01 mg/mL to about 2 mg/mL, about 0.01 mg/mL to about 5 mg/mL, about 0.01 mg/mL to about 10 mg/mL, about 0.1 mg/mL to about 0.5 mg/mL, about 0.1 mg/mL to about 1 mg/mL, about 0.1 mg/mL to about 1.25 mg/mL, about 0.1 mg/mL to about 1.5 mg/mL, about 0.1 mg/mL to about 1.75 mg/mL, about 0.
  • the lipid vesicle composition comprises the anionic polymer material in an amount of about 0.01 mg/mL, about 0.05 mg/mL, about 0.1 mg/mL, about 0.5 mg/mL, about 1 mg/mL, about 1.25 mg/mL, about 1.5 mg/mL, about 1.75 mg/mL, about 2 mg/mL, about 5 mg/mL, or about 10 mg/mL.
  • Peptide Composition 2 In one aspect, provided herein, is a lipid vesicle composition comprising:
  • lipid vesicles each comprising a lipid bilayer comprising vesicle forming lipids, wherein the vesicle forming lipids are present in an amount of from about 2% to about 20%;
  • composition further comprises: a PEG fatty acid ester in an amount of from about 0.1% to about 2%; a polysorbate in an amount of from about 0.5 % to about 3%; and a sorb ate ester in an amount of from about 0.1% to about 2%.
  • the oil component is present in an amount of from about 2.5% to about 20 %.
  • the lipid vesicle composition comprises the peptide antagonist in an amount of about 0.1 mg/mL to about 0.5 mg/mL, about 0.1 mg/mL to about 1 mg/mL, about 0.1 mg/mL to about 2 mg/mL, about 0.1 mg/mL to about 3 mg/mL, about 0.1 mg/mL to about 4 mg/mL, about 0.1 mg/mL to about 5 mg/mL, about 0.1 mg/mL to about 10 mg/mL, about 0.1 mg/mL to about20 mg/mL, about 0.1 mg/mL to about50 mg/mL.
  • the lipid vesicle composition comprises the peptides antagonist in an amount of about 0.1 mg/mL, about 0.5 mg/mL, about 1 mg/mL, about2 mg/mL, about 3 mg/mL, about 4 mg/mL, about 5 mg/mL, about 10 mg/mL, about 20 mg/mL, or about 50 mg/mL. In some embodiments, the lipid vesicle composition comprises the peptides antagonist in an amount of about 1 mg/mL, about 2 mg/mL, about 3 mg/mL, about 4 mg/mL, or about 5 mg/mL.
  • the lipid vesicle composition further comprises viscosity enhancing agents in an amount of from about 0.5% to about 5%.
  • the viscosity enhancing agents comprise one or more of a fatty alcohol, a wax, a fatty ester of glycerol, or any combination thereof.
  • the PEG fatty acid ester comprises PEG4-dilaurate.
  • the polysorbate is polysorbate 80.
  • the sorbate ester is sorbitan palmitate.
  • the lipid vesicle composition further comprises an anionic polymer material in an amount of from about 0.01 mg/mL to about 10 mg/mL entrapped in the lipid bilayer, the oil-in-water emulsion, or a combination thereof.
  • the lipid vesicle composition comprises the anionic polymer material in an amount of about 0.01 mg/mL to about0.05 mg/mL, about 0.01 mg/mL to about 0.1 mg/mL, about 0.01 mg/mL to about 0.5 mg/mL, about 0.01 mg/mL to about 1 mg/mL, about 0.01 mg/mL to about 1.25 mg/mL, about 0.01 mg/mL to about 1.5 mg/mL, about 0.01 mg/mL to about 1.75 mg/mL, about 0.01 mg/mL to about 2 mg/mL, about 0.01 mg/mL to about 5 mg/mL, about 0.01 mg/mL to about 10 mg/mL, about 0.1 mg/mL to about 0.5 mg/mL, about 0.1 mg/mL to about 1 mg/mL, about 0.1 mg/mL to about 1.25 mg/mL, about 0.1 mg/mL to about 1.5 mg/mL, about 0.1 mg/mL to about 1.75 mg/mL, about 0.
  • the lipid vesicle composition comprises the anionic polymer material in an amountof about 0.01 mg/mL, about0.05 mg/mL, about 0.1 mg/mL, about 0.5 mg/mL, about 1 mg/mL, about 1.25 mg/mL, about 1.5 mg/mL, about 1.75 mg/mL, about 2 mg/mL, about 5 mg/mL, or about 10 mg/mL.
  • Peptide Composition 3 In one aspect, provided herein, is a lipid vesicle composition comprising:
  • lipid vesicles each comprising a lipid bilayer comprising vesicle forming lipids, wherein the vesicle forming lipids are present in an amount of from about 5% to about 20%;
  • composition further comprises: and a fatty amide derived propylene glycol -diammonium phosphate ester in an amountof from about 1% to about 10%; a PEG ether of a fatty alcohol in an amount of from about 0.1% to about 3%; a polysorbate in an amount of from about 0.5 % to about 3%; and a sorb ate ester in an amount of from about 0.1% to about 2%.
  • the oil component is present in an amount of from about 2.5% to about 20 %.
  • the lipid vesicle composition comprises the peptide antagonist in an amount of about 0.1 mg/mL to about 0.5 mg/mL, about 0.1 mg/mL to about 1 mg/mL, about 0.1 mg/mL to about 2 mg/mL, about 0.1 mg/mL to about 3 mg/mL, about 0.1 mg/mL to about 4 mg/mL, about 0.1 mg/mL to about 5 mg/mL, about 0.1 mg/mL to about 10 mg/mL, about 0.1 mg/mL to about20 mg/mL, about 0.1 mg/mL to about50 mg/mL.
  • the lipid vesicle composition comprises the peptides antagonist in an amount of about 0.1 mg/mL, about 0.5 mg/mL, about 1 mg/mL, about2 mg/mL, about 3 mg/mL, about 4 mg/mL, about 5 mg/mL, about 10 mg/mL, about 20 mg/mL, or about 50 mg/mL. In some embodiments, the lipid vesicle composition comprises the peptides antagonist in an amount of about 1 mg/mL, about 2 mg/mL, about 3 mg/mL, about 4 mg/mL, or about 5 mg/mL.
  • the lipid vesicle composition further comprises viscosity enhancing agents in an amount of from about 0.5% to about 5%.
  • the viscosity enhancing agents comprise one or more of a fatty alcohol, a wax, a fatty ester of glycerol, or any combination thereof.
  • the polysorbate is polysorbate 80.
  • the sorbate ester is sorbitan palmitate.
  • the PEG ether of the fatty alcohol is diethylene glycol monooleyl ether.
  • the lipid vesicle composition further comprises an anionic polymer material in an amount of from about 0.01 mg/mL to about 10 mg/mL entrapped in the lipid bilayer, the oil-in-water emulsion, or a combination thereof.
  • the lipid vesicle composition comprises the anionic polymer material in an amount of about 0.01 mg/mL to about0.05 mg/mL, about 0.01 mg/mL to about 0.1 mg/mL, about 0.01 mg/mL to about 0.5 mg/mL, about 0.01 mg/mL to about 1 mg/mL, about 0.01 mg/mL to about 1.25 mg/mL, about 0.01 mg/mL to about 1.5 mg/mL, about 0.01 mg/mL to about 1.75 mg/mL, about 0.01 mg/mL to about 2 mg/mL, about 0.01 mg/mL to about 5 mg/mL, about 0.01 mg/mL to about 10 mg/mL, about 0.1 mg/mL to about 0.5 mg/mL, about 0.1 mg/mL to about 1 mg/mL, about 0.1 mg/mL to about 1.25 mg/mL, about 0.1 mg/mL to about 1.5 mg/mL, about 0.1 mg/mL to about 1.75 mg/mL, about 0.
  • the lipid vesicle composition comprises the anionic polymer material in an amountof about 0.01 mg/mL, about0.05 mg/mL, about 0.1 mg/mL, about 0.5 mg/mL, about 1 mg/mL, about 1.25 mg/mL, about 1.5 mg/mL, about 1.75 mg/mL, about 2 mg/mL, about 5 mg/mL, or about 10 mg/mL.
  • lipid vesicle compositions are contemplated for cosmetic uses in a subject, for indications including but not limited to for preventing, reducing, or both, the appearance of skin discoloration due to pigmentation .
  • the lipid vesicle compositions provided herein are contemplated for pharmaceutical use in a subject, for indications including but not limited to: the skin discoloration, e.g., pigmentation or hyperpigmentation, comprises melanin hyperpigmentation, chloasma, melasma, age spots, freckles, or a combination thereof.
  • the skin discoloration results from a disorder, e.g., an adrenal disorder.
  • the MC1R peptide antagonists of the disclosure are used to treat skin discoloration caused by Addison's disease.
  • the subject is a mammal.
  • the mammal is a human.
  • the human subject is a pediatric or adult subject, of any age.
  • the present disclosure also relates to methods for using cosmetic or pharmaceutical compositions comprising a peptide antagonist.
  • the disclosure comprises a method of preventing or reducing skin discoloration in a subject, the method comprising administering a lipid composition to said subject, wherein the lipid vesicle composition comprises: (a) lipid vesicles each comprising a lipid bilayer comprising vesicle forming lipids, (b) an oil -in-water emulsion entrapped in the lipid vesicles, and stabilized by one or more surfactants; (c) a peptide antagonist of a melanocortin 1 receptor entrapped in the lipid bilayer and/or the oil -in-water emulsion.
  • the skin discoloration is melanin hyperpigmentation, chloasma, melasma, age spots, freckles, or a combination thereof.
  • the disclosure relates to methods for using the cosmetic or pharmaceutical composition to prevent or reduce skin discoloration, comprising applying an effective amount of the cosmetic or pharmaceutical composition to the skin of the subject.
  • the skin discoloration comprises melanin hyperpigmentation, post-inflammatory hyperpigmentation, chloasma, melasma, age spots (e.g., liver spots, senile lentigines, solar lentigines, sunspots), freckles, or a combination thereof.
  • the skin discoloration results from a disorder, e.g., an adrenal disorder.
  • the MC1R peptide antagonists of the disclosure are used to treat skin discoloration caused by Addison's disease.
  • the cosmetic or pharmaceutical composition is used to diminish or visible remove
  • the cosmetic or pharmaceutical composition diminish color in dark spots. In some embodiments, the cosmetic or pharmaceutical composition provides a more even tone across the face of the subject.
  • the lipid vesicle composition is topically applied to a subject.
  • Topical application as referred to herein can refer to application onto one or more surface, e.g., keratinous tissue.
  • the topical composition is administered to the skin of a subject.
  • the skin is the facial skin of the subject.
  • Topical application may relate to direct application to the desired area.
  • a topical cosmetic or pharmaceutical composition or preparation can be applied by, e.g., pouring, dropping, or spraying, when present as a liquid or aerosol composition; smoothing, rubbing, spreading, and the like, when in ointment, lotion, cream, gel, or a like composition; dusting, when a powder; or by any other appropriate means.
  • the lipid vesicle composition is formulated in a form suitable for topical application.
  • the lipid vesicle composition is formulated as a cream, a lotion, a suspension, or an emulsion.
  • the lipid vesicle composition is formulated as a cream.
  • the lipid vesicle composition is formulated as a lotion.
  • the lipid vesicle composition is formulated as a suspension.
  • the subject uses or is treated with a topical application comprising an effective amount of the lipid vesicle composition one time or more during a course of usage or treatment, e.g., 1 -3 times per day, 1 - 21 times per week, 1 time per day, 2 times per day, or 3 times per day.
  • a subject uses oris treated with an effective amount of the lipid vesicle composition about 1 time per week to about 12 times per week.
  • a subject uses oris treated with an effective amount of the lipid vesicle composition at least about 1 time per week.
  • a subject uses oris treated with an effective amount of the lipid vesicle composition at most about 12 times per week. In some embodiments, a subject uses or is treated with an effective amount of the lipid vesicle composition about 1 time per week to about 2 times per week, about 1 time per week to about 3 times per week, about 1 time per week to about 4 times per week, about 1 time per week to about 5 times per week, about 1 time per week to about 6 times per week, about 1 time per week to about 7 times per week, about 1 time per week to about 8 times per week, about 1 time per week to about 9 times per week, about 1 time per week to about 10 times per week, about 1 time per week to about 11 times per week, about 1 time per week to about 12 times per week, about 2 times per week to about 3 times per week, about 2 times per week to about 4 times per week, about 2 times per week to about 5 times per week, about 2 times per week to about 6 times per week, about 2 times per week to about 7 times per week, about 2 times
  • a subject uses oris treated with an effective amount of the lipid vesicle composition about 1 time per week, about 2 times perweek, about 3 times per week, about 4 times per week, about 5 times perweek, about 6 times perweek, about 7 times perweek, about 8 times perweek, about 9 times perweek, about 10 times perweek, about 11 times perweek, about 12 times perweek, about 13 times perweek, or about 14 times perweek.
  • one or more layers of a lipid vesicle composition of the disclosure is applied to the skin of the subject at a given time.
  • a subsequent layer may be applied after a previous layer of the lipid vesicle composition is fully absorbed into the skin of the subject.
  • the lipid vesicle composition may take a couple of seconds (e.g., one second, two seconds, three second, five seconds, ten seconds, fifteen seconds, thirty seconds, etc.) to fully absorb into the skin of the subject.
  • one, two, three, four, five, six, or seven layers of the lipid vesicle composition is applied to the skin of the subject at a given time.
  • the lipid vesicle composition is applied to the skin of the subject one or more times a day (e.g., 1-3 times per day, 1 time per day, 2 times per day, 3 times per day, etc.).
  • the lipid vesicle composition is applied to the skin of the subject one or more times a week (e.g., 1-21 times per week, 1-14 times per week, 1- 7 times per week, etc.). In some embodiments, the lipid vesicle composition is applied to the skin of the subject daily. In some embodiments, one or more layers of the lipid vesicle composition is applied to the skin of the subject once a day for one or more days. In some embodiments, two or more layers of the lipid vesicle composition is applied to the skin of the subject once a day for one or more days. In some embodiments, three or more layers of the lipid vesicle composition is applied to the skin of the subject once a day for one or more days.
  • one or more layers of the lipid vesicle composition is appliedto the skin of the subject twice a day for one or more days. In some embodiments, two or more layers of the lipid vesicle composition is appliedto the skin of the subject twice a day for one or more days.
  • three or more layers of the lipid vesicle composition is applied to the skin of the subject twice a day for one or more days.
  • the lipid vesicle composition is applied to the skin of the subject for at least one day, two days, three days, four days, five days, six days, one week, two weeks, three weeks, one month, two months, three months, six months, one year.
  • the lipid vesicle composition is applied to the skin of the subject for more than one day, two days, three days, four days, five days, six days, one week, two weeks, three weeks, one month, two months, three months, six months, nine months, or one year.
  • one or more layers of the lipid vesicle composition is applied to the skin of the subject twice a day for several days, and thereafter is applied three times a day.
  • five layers of the lipid vesicle composition is applied to the skin of the subject twice a day for five days (e.g., morning and night), and thereafter one to three layers of the lipid vesicle composition is applied to the skin of the subject three times a day (e.g., morning, noon and night).
  • a lipid vesicle composition ofthe disclosure is administered to a subject, for indications including but not limited to: prevention or temporary improvement of the appearance of skin discoloration e.g., dark spots, melanin hyperpigmentation, post-inflammatory hyperpigmentation, chloasma, melasma, age spots (e.g., liver spots, senile lentigines, solar lentigines, sunspots), freckles, or a combination thereof.
  • a lipid vesicle composition ofthe disclosure is administered to a subject, for indications including but not limited to: improvement of the appearance of skin discoloration.
  • a lipid vesicle composition of the disclosure is used with other products, including, but not limited to sunscreen, moisturizers, face creams (e.g., BB cream, CC cream, night cream, etc.), mists, foundations, concealers, highlighters, primers, etc.
  • a topical cosmetic composition of the disclosure is self -applied by a subject. In some embodiments, a topical cosmetic composition of the disclosure is not self- applied by a subject. In some embodiments, a topical cosmetic composition of the disclosure is administered by a subject. In certain embodiments, including pharmaceutical embodiments, a cosmetic or pharmaceutical composition of the disclosure is applied or administered by a medical professional, e.g., in a medical office setting.
  • compositions of the disclosure as described above are prepared by mixing oil components of the oil-in-water emulsion with aqueous components of the oil -in-water emulsion wherein either the oil components or aqueous components of the oil -in-water emulsion comprises one or more surfactants for emulsification of the oil component with the aqueous component of the oil-in-water emulsion.
  • the surfactant is mixed with the aqueous component and added to the oil for formation of an emulsion.
  • the oil -in water emulsion is then mixed with the solubilized vesicle-forming lipid and, if added, other lipid components under mixing conditions effective to form the lipid vesicles (e.g., multisomes).
  • one or more penetration enhancing agents and the one or more compounds are added to oil component of the oil -in-water emulsion, to the aqueous component of the oil-in- water emulsion or both.
  • the one or more penetration enhancing agents and/or the one or more compounds can be added to the lipid component.
  • a method of preparing a lipid vesicle composition comprising a) preparing an oil-in-water emulsion comprising an active ingredient, by mixing oil components of the oil-in-water emulsion with aqueous components of the oil-in-water emulsion; b) solubilizing vesicle forming lipids in an acceptable solvent other than water; c) adding the oil-in-water emulsion to the solubilized vesicle forming lipids; and d) mixing the oil-in-water emulsion and the solubilized vesicle forming lipids under mixing conditions effective to form the lipid vesicles comprising a lipid bilayer comprising vesicle forming lipids, and an oil-in-water emulsion entrapped in the lipid vesicles.
  • a method of preparing a lipid vesicle composition comprising: a) preparing an oil-in-water emulsion comprising the peptide antagonist of a melanocortin 1 receptor, by mixing oil components of the oil -in-water emulsion with aqueous components of the oil-in-water emulsion, wherein the oil components and/or the aqueous components of the oil-in-water emulsion comprises the one or more surfactants; b) solubilizing vesicle forming lipids in an acceptable solvent other than water; c) adding the oil-in-water emulsion to the solubilized vesicle forming lipids; and d) mixing the oil-in-water emulsion and the solubilized vesicle forming lipids under mixing conditions effective to form the lipid vesicles comprising a lipid bilayer comprising vesicle
  • the active ingredient is one or more peptides provided herein.
  • the one or more peptides comprises an amino acid sequence at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% sequence homology to the amino acid sequence of any one of SEQ ID NO: 1 -90. In some embodiments, the one or more peptides comprises an amino acid sequence identical to the amino acids of any one of SEQ ID NO: 1 -90. In some embodiments, the one or more peptides comprises an amino acid sequence at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% sequence homology to the amino acid sequence of any one of SEQ ID NO: 4-19 or 44-65.
  • the one or more peptides comprises an amino acid sequence identical to the amino acids of any one of SEQ ID NO: 4-19 or 44-65.
  • the active ingredient is an anionic polymer material provided herein.
  • the lipid vesicle may comprise of one or more active ingredients.
  • the one or more active ingredients comprise one or more peptides.
  • the one or more active ingredients comprise an anionic polymer material.
  • the one or more active ingredients comprise one or more peptides and an anionic polymer material.
  • the anionic polymer material is hyaluronic acid.
  • the method further comprises adding one or more of the additional components provided herein (e.g., penetration enhancing agents, viscosity enhancing agents, etc.).
  • mixing oil components of the oil -in-water emulsion with aqueous components of the oil -in-water emulsion vesicles of step a) and/or the mixing conditions of step e) comprises using agitation such as homogenization or emulsification, or micro -emulsion techniques which do not involve agitation.
  • the mixing comprises high pressure homogenizing.
  • the high pressure homogenizing provides relatively precise control over the composition of the lipid vesicles. High pressure homogenizing is suitable for small molecules and peptides or proteins that are resistant to shearing.
  • the composition that is formed is any one of the lipid vesicle compositions described herein.
  • other lipid components are added at any one of the steps.
  • Peptide antagonists of the disclosure having amino acid sequences as set forth in SEQ ID NOS: 11, 16, 25, 27, 32, 35, and 40 (each having a C-terminal -NH2) were screened at five concentrations, as well as a positive control, SEQ ID NO: 1 (Melanostatine-5). Inhibition curves were obtained by determining the reduction of bound cr-MSH in the presence of an amount of, and/or by increasing amounts of, each peptide antagonist. Non-specific binding of peptide was estimated separately in the presence of a vast excess (1 mM) of unlabeled cr-MSH, which occupies all the available receptor binding sites.
  • the IC50 values were determined (Eurofins or AAT Bioquest IC 50 calculator). As shown in Table 4, the IC 50 value for the Melanostatine-5 control (SEQ ID NO: 1) was 120 nM. The test peptides had IC 50 values ranging from below 74 nm (markedly below Melanostatine-5), to greater than 1 uM. Table 4. Binding of Peptide Antagonists to MC1R
  • Group B peptide antagonists of the disclosure having amino acid sequences as set forth in Table 2 are screened at 10 mM, as well as one or more positive control selected from SEQ ID NO: 1 (Melanostatine-5), the 132-amino acid natural MC1R antagonist Agouti Signaling Protein (ASIP) (SEQ ID NO: 91), mature ASIP (SEQ ID NO: 92), a fragment of ASIP, e.g., ASIP 107-132 (SEQ ID NO: 2), or a variant of ASIP or fragment thereof, e.g., ASIP-YY (SEQ ID NO: 93), ASIP-YY 107-132 subdomain (SEQ ID NO: 3), or ASIP-YY 93-132 subdomain (SEQ ID NO: 94).
  • ASIP Agouti Signaling Protein
  • Inhibition curves are obtained by determining the reduction of bound cr-MSH in the presence of an amount of, and/or by increasing amounts of, each peptide antagonist. Non-specific binding of peptide is stimulated separately in the presence of a vast excess (1 mM) of unlabeled cr-MSH, which occupies all the available receptor binding sites.
  • formulations comprising MC1R peptide antagonists of the disclosure, e.g., any peptide having an amino as set forth in Table 1 and Table 2, are evaluated in comparison to controls (e.g., no treatment or treatment with agonist cr-MSH) and positive controls for lightening (e.g., formulations comprising a peptide having a sequence set forth in any of SEQ ID NOS: 1-3, 91, 92, 93 and 94, or a small molecule tyrosinase inhibitor such as Kojic Acid).
  • controls e.g., no treatment or treatment with agonist cr-MSH
  • positive controls for lightening e.g., formulations comprising a peptide having a sequence set forth in any of SEQ ID NOS: 1-3, 91, 92, 93 and 94, or a small molecule tyrosinase inhibitor such as Kojic Acid.
  • Pigmentation is evaluated over the course of 2-3 weeks using a tristimulus chromometer to measure brightness (L*) in MelanoDerm tissue produced with normal human melanocytes from Black, Asian, or Caucasian donors. In parallel to measurements taken with the chromameter, total melanin content of tissues is also quantified.
  • Multisomes Biphasic vesicles with multiple/synergistic penetration enhancers (Multisomes) were formulated using the procedure described herein.
  • the multisomes may comprise a peptide antagonist of a melanocortin 1 receptor, such as those described herein.
  • the general procedure for multisome preparation was as follows:
  • Some of the multisome formulations that were prepared comprised a peptide.
  • the peptide was a melanocortin receptor 1 antagonist.
  • the melanocortin receptor 1 antagonist was mixed into the water phase in the procedure described herein.
  • the mixture was intermittently vortexedand heated for 5 sec/5 sec for 8-10 cycles until a uniform creamy lotion formed.
  • FIG. 1 A pictorial representation of this process is shown in FIG. 1.
  • FIG. 2. A flow chart of this exemplary process is shown in FIG. 2.
  • the following methods are used to characterize the formulations prepared using the method of Example 4 well as the performance of the formulation.
  • the formulations prepared using Example 4 contain peptide antagonist of melanocortin 1 receptors, such as any one of SEQ ID NOs: 4-90.
  • Size (hydrodynamic diameter), polydispersity index and zeta (V) potential measurements are carried out on formulations prepared with non-labelled peptides using a dynamic light scattering (DLS) instrument, for example, the Zetasizer NanoZS (Malvern Instruments, Worcestershire, UK). Aliquots of formulations are assessed for particle size distribution and subsequently diluted with water for zeta potential measurements. Measurements are carried out in triplicates.
  • DLS dynamic light scattering
  • diffusion cells for example, 9mm diameter Bronaugh -type teflon flow through diffusion cells with an exposed surface area of about 0.6 cm 2 .
  • the cell holder is maintained at about 32°C by a circulating water bath heater.
  • a perfusion fluid is used in the diffusion cell, for example, degassed phosphate-buffered saline (PBS) buffer with 0.05% sodium- azide pH 7.2, maintained at 37°C, with a flow rate of 1 mL/h.
  • PBS degassed phosphate-buffered saline
  • Skin samples are removed from the freezer and cut into about 1cm x 1cm square pieces and mounted into the diffusion cells epidermis facing up.
  • Application is performed for periodically and transdermal fractions are collected into tissue culture tubes using a programmed fraction collector .
  • Transdermal fraction analysis The transdermal fractions are collected periodically for analysis, for example hourly for 24h. The samples are sent for analysis to a laboratory.
  • the skin samples from the diffusion cell study are cleansed by the usual protocol to remove residual bound cream, i.e. after the skin samples are removed from the diffusion cells and washed with water, and patted dry with a kimwipe.
  • the cleansed skin discs are stored at about - 20°C.
  • Multisome formulation optimization and characterization All formulations are lotion or cream consistency suitable for topical application. The formulations are physically stable showing no separation, sedimentation or other signs of stability issues for >1 mo of storage at about 4°C.
  • formulations are shown to be poly disperse with ranging vesicle sizes as shown in light microscopic images.
  • the microscopic observations confirm the formation of multisomes with the typical biphasic vesicle morphology and the uniform distribution of vesicles throughout the formulation for one or more peptide-containing and blank (no peptide) formulations.
  • Each of the one or more peptide-containing formulations are similar with respect to size distribution compared to their respective blank formulations, but overall, the blank formulations have narrower size distribution compared to the peptide formulations. Zetasizer data show consistent results with the microscopic observations, typical of multisomes.
  • Lipid vesicle formulations are prepared with one or more peptide and applied to skin samples. Blank versions of each formulation are prepared as controls, and a solution of the one or more peptides in water is prepared as an additional control. Each formulation is tested in triplicates and blank formulations, skin without any formulations applied, and the one or more peptide solution as free, non-encapsulated peptides are used for background fractions for the analysis.
  • transdermal fractions are collected for further analysis by mass spectrometry by a laboratory.
  • the total amount (Qt (24h)) of the peptides delivered through the diameter skin disk applied in the diffusion cells and the delivery rates for each formulation are measured.
  • a lipid vesicle formulation of one or more peptides of the disclosure is tested for safety and efficacy for facial application of dark spots in a randomized, double-blind human clinical trial. An amount of the lipid vesicle formulation, or placebo (blank lipid vesicle) is applied to dark spots on the faces of individuals on Day 1.
  • Secondary outcome Percentage of Participants Achieving Satisfied or Very Satisfied by Subject Assessment of Satisfaction of Appearance of Dark Spots (participant assessment).
  • Peptide antagonists of the disclosure having amino acid sequences as set forth in SEQ ID NOS: 11, 16, 25, 27, 32, 35, and 40 were screened at five concentrations. Inhibition curves were obtained by determining the reduction of bound cr-MSH in the presence of an amount of, and/or by increasing amounts of, each peptide antagonist.
  • Example 8 Evaluation of the Efficacy of Peptides for Facial Application
  • a lipid vesicle formulation comprising a peptide of Example 7 was tested for efficacy for facial application of dark spots in a human clinical trial of 15 subjects.
  • 50% of the subjects were Caucasian and 50% were non-Caucasian, including Hispanic, Asian, African American, East Indian, native America, and Middle Easter ethnicities.
  • the skincare regiment for the subjects included the use of a cleaner on the face, followed by application of the lipid vesicle formulation, and further followed by the application of a moisturizer to the skin of the face.
  • the skincare regiment was followed twice a day, in the morning and at night.
  • the morning skincare regiment further included the application of a sunscreen with SPF50+to the skin of the face.
  • the skincare routine was followed for two weeks.
  • the skin of the faces of the subjects were analyzed after two weeks using the Visia® CR skin analysis imaging system.
  • An exemplary resultfrom a subject is shown in FIG. 3. As shown, the dark spots on the face of the subject was visibly reduced after two weeks.

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