EP1349564A2 - Lokale applikation von peptiden zur regulation der zellproliferation und zelldifferenzierung - Google Patents

Lokale applikation von peptiden zur regulation der zellproliferation und zelldifferenzierung

Info

Publication number
EP1349564A2
EP1349564A2 EP01946566A EP01946566A EP1349564A2 EP 1349564 A2 EP1349564 A2 EP 1349564A2 EP 01946566 A EP01946566 A EP 01946566A EP 01946566 A EP01946566 A EP 01946566A EP 1349564 A2 EP1349564 A2 EP 1349564A2
Authority
EP
European Patent Office
Prior art keywords
lipid
proliferation
peptide
skin
hair
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP01946566A
Other languages
English (en)
French (fr)
Inventor
Michael F Holick
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Publication of EP1349564A2 publication Critical patent/EP1349564A2/de
Withdrawn legal-status Critical Current

Links

Classifications

    • 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
    • C07K14/575—Hormones
    • C07K14/635—Parathyroid hormone, i.e. parathormone; Parathyroid hormone-related peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/29—Parathyroid hormone, i.e. parathormone; Parathyroid hormone-related peptides
    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Liposomes
    • A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers
    • A61K9/1272—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q7/00—Preparations for affecting hair growth
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q7/00—Preparations for affecting hair growth
    • A61Q7/02—Preparations for inhibiting or slowing hair growth

Definitions

  • This invention relates to the regulation of cell differentiation and proliferation, e.g., for treating hyperproliferative skin disorder, such as psoriasis, for enhancing wound healing, for stimulating hair growth, and inhibiting hair growth by topical administration of parathyroid hormone (PTH), parathyroid related peptide (PTHrP), or fragment, analog or derivative thereof, and salts thereof, encapsulated by particular liposomes.
  • hyperproliferative skin disorder such as psoriasis
  • PTHrP parathyroid hormone
  • PTHrP parathyroid related peptide
  • salts thereof encapsulated by particular liposomes.
  • U.S. Pat. Nos. 5,527,772, 5,840,690 and 6,066,618 describe methods of inhibiting proliferation and enhancing differentiation of mammalian cells, inducing proliferation of mammalian cells, enhancing wound healing, and stimulating hair growth using a peptide which has a 10% or greater homology to a region of human PTH or human PTHrP.
  • Certain fragments and analogs e.g. PTH (1-34), PTH (3-34) and PTHrP (1-34)
  • PTHrP 1-34
  • Other fragments and analogs e.g. PTH (7-34) and PTHrP
  • the agonists are useful for treatment of hyperproliferative skin diseases such a psoriasis and the antagonists are useful for wound healing, particularly wounds of the skin, enhancing or maintaining hair growth, particularly following chemotherapeutic treatment of a mammal, and stimulating epidermal regrowth.
  • Methods of administration include oral, nasal, intravenous, subcutaneous, parenteral and intraperitoneal administration.
  • the peptides may be administered by subcutaneous pumps, patches, tapes, or by liposomal carriers.
  • PTH and PTHrP analogs and derivatives thereof have been made. See U.S. Pat. Nos. 4,086,196, 4,423,037, 4,771,124, 4,833,125, 4,968,669, 5,001,223, 5,087,562, 5,093,233, 5,116,952, 5,149,779, 5,171,670,
  • Active vitamin D compounds are useful for treating hype roliferative skin diseases and other conditions.
  • a large number of such active vitamin D compounds are known. See U.S. Patent Nos. 5,457,217, 5,414,098, 5,384,313, 5,373,004, 5,371,249, 5,430,196, 5,260,290, 5,393,749, 5,395,830, 5,250,523,
  • the invention provides two important therapeutic methods one involving inhibition of cell proliferation and enhancement of skin cell differentiation (the agonist activity), which is useful in the treatment of psoriasis, ichthyosis, actinic keratoses, skin cancer, inhibiting hair growth or preventing hair regrowth.
  • a second method involves enhancement of cell proliferation (the antagonist activity), which is useful in wound healing, stimulating epidermal regrowth and hair growth.
  • the invention provides methods for enhancing wound healing and hair growth based on in vivo wound healing activity or in vitro or in vivo hair growth activity rather than strict agonist or antagonist activity in vitro.
  • the first method of the invention generally involves inhibiting proliferation and enhancing differentiation of mammalian skin cells by contacting the cell with a liposomal preparation comprising a peptide (preferably at least 3, and more preferably at least 8, amino acids long) which has 10% or greater (more preferably, 50% or greater, and most preferably 75% or greater) sequence identity with a region (preferably within the amino- terminal 34 amino acid region) of human PTH or human PTHrP, and which is capable of inhibiting proliferation or enhancing the differentiation in vitro of cultured human keratinocytes; or in vivo in mouse skin by inhibiting skin cell proliferation or hair cycle progression or hair growth.
  • a liposomal preparation comprising a peptide (preferably at least 3, and more preferably at least 8, amino acids long) which has 10% or greater (more preferably, 50% or greater, and most preferably 75% or greater) sequence identity with a region (preferably within the amino- terminal 34 amino acid region) of human PTH or human PTHrP, and which is capable of inhibiting proliferation or enhancing
  • the peptide is hPTH (1-84), hPTH (1-34), hPTH (3-34), hPTHrP (1-34), hPTHrP (1-141), hPTHrP (1-139) or hPTHrP (1-173).
  • This method has particular application in the treatment of hyperproliferative skin disorders such as psoriasis.
  • the method may also be useful in the treatment of certain skin cancers, by the inhibition of cancer cell proliferation and by the induction of differentiation and inhibition of hair growth.
  • the second method of the invention generally involves enhancing proliferation of mammalian skin cells by contacting the skin cells with a liposomal preparation comprising a peptide (preferably at least 3, and more preferably at least 8, amino acids long) which has 10% or greater (more preferably, 50% or greater, and most preferably 75% or greater) sequence identity with a region (preferably within the amino-terminal 34 amino acid region) of hPTH or hPTHrP, and which is capable of blocking the differentiation or the inhibition of proliferation in vitro of cultured human keratinocytes by PTH (1-34) or l,25(OH)2D3 or PTHrP (1-34); or in vivo in mouse skin by stimulating skin cell proliferation or accelerating hair cycle progression or stimulating hair growth.
  • a liposomal preparation comprising a peptide (preferably at least 3, and more preferably at least 8, amino acids long) which has 10% or greater (more preferably, 50% or greater, and most preferably 75% or greater) sequence identity with a region (preferably within the amino-termin
  • the peptide is PTH (7-34), hPTH (5-34) or hPTHrP (5-34).
  • proliferation of mammalian skin cells is enhanced by contacting the cell or wound with a liposomal preparation comprising a peptide (preferably at least 3, and more preferably at least 8, amino acids long) which has 10% or greater (more preferably, 50% or greater, and most preferably, 75% or greater) sequence identity with a region (preferably, within the amino-terminal 34 amino acid region) of hPTH or hPTHrP, and which is capable of enhancing wound healing in an in vivo skin punch assay.
  • a liposomal preparation comprising a peptide (preferably at least 3, and more preferably at least 8, amino acids long) which has 10% or greater (more preferably, 50% or greater, and most preferably, 75% or greater) sequence identity with a region (preferably, within the amino-terminal 34 amino acid region) of hPTH or hPTHrP, and which is capable of enhancing wound healing
  • the peptide is hPTH (1-84), hPTH (1-34), hPTH (7-34), hPTH (5-34), hPTH (5-36), hPTHrP (1-34), or hPTHrP (7-34).
  • these related methods have particular application in the enhancement of wound healing and also have applications in the promotion of skin growth in patients with burns or skin ulcerations as well as in the stimulation of epidermal regrowth in people who have decreased epidermal cell proliferation due to aging.
  • Hair growth is stimulated by administering to a mammal a liposomal preparation comprising a peptide (preferably at least 3, and more preferably at least 8, amino acids long) which has 10% or greater (more preferably, 50% or greater, and most preferably, 75% or greater) sequence identity with a region (preferably, within the amino-terminal 34 amino acid region) of hPTH or hPTHrP, and which is capable of stimulating hair growth in vitro or in vivo.
  • the peptide is hPTH (7-34), hPTH (5- 34) or hPTH (5-36). This method has applications in the promotion of new hair growth or stimulation of the rate of hair growth, e.g., following chemotherapeutic treatment or for treating a form of alopecia, e.g., male or female pattern baldness.
  • the invention relates to a method of inhibiting proliferation or enhancing differentiation of a mammalian skin or hair cell, the method comprising topically admimstering to the mammalian skin or hair cell in need of inhibited proliferation or enhanced differentiation with a proliferation-inhibiting or differentiation-enhancing amount of a peptide or a salt or derivative thereof encapsulated within a liposome, wherein the peptide is at least 3 amino acids long, has at least 10% sequence identity with the 34 amino acid N-terminal region of hPTH or hPTHrP, and is capable of inhibiting proliferation or enhancing differentiation in vitro of cultured human keratinocytes; or in vivo in mouse skin by inhibiting skin cell proliferation or hair cycle progression or hair growth; wherein the liposome comprises at least two distinct lipids, a primary lipid and a secondary lipid, the primary lipid constituting the greatest proportion, by weight, of any single lipid material forming the bilayers of said vesicle, the primary lipid being selected
  • the invention also relates to a method of inhibiting proliferation or enhancing differentiation of a skin or hair cell of a mammal, comprising administering to the mammal in need thereof a proliferation-inhibiting or differentiation-enhancing amount of a peptide or a salt or derivative thereof and an active vitamin D compound, wherein the peptide is at least 3 amino acids long, has at least 10% sequence identity with the 34 amino acid N- terminal region of hPTH or hPTHrP, and is capable of inhibiting proliferation or enhancing differentiation in vitro of cultured human keratinocytes, or in vivo in mouse skin by inhibiting skin cell proliferation or hair cycle progression or hair cell growth.
  • the invention also relates to a method of inducing proliferation of a mammalian skin or hair cell, the method comprising topically administering to the mammalian skin or hair cell in need of proliferation with a proliferation- inducing amount of a peptide or a salt or derivative thereof encapsulated within a liposome, wherein the peptide is at least 3 amino acids long, has at least 10% sequence identity with the 34 amino acid N-terminal region of hPTH or hPTHrP, and is capable of blocking the inhibition of proliferation or stimulation of differentiation in vitro of cultured human keratinocytes by PTH (1-34), l,25(OH)2D3 or PTHrP (1-34), or in vivo in mouse skin by stimulating skin cell proliferation or accelerating hair cycle progression or stimulating hair growth; wherein said liposome comprises at least two distinct lipids, a primary lipid and a secondary lipid, the primary lipid constituting the greatest proportion, by weight, of any single lipid material forming the bilayers of said
  • the invention also relates to a composition
  • a composition comprising a proliferation- inhibiting or differentiation-enhancing amount of a peptide or a salt or derivative thereof encapsulated within a liposome, wherein the peptide is at least 3 amino acids long, has at least 10% sequence identity with the 34 amino acid N-terminal region of hPTH or hPTHrP, and is capable of inhibiting proliferation or enhancing differentiation in vitro of cultured human keratinocytes, or in vivo in mouse skin by inhibiting skin cell proliferation or hair cycle progression or hair growth; wherein said liposome comprises at least two distinct lipids, a primary lipid and a secondary lipid, the primary lipid constituting the greatest proportion, by weight, of any single lipid material forming the bilayers of said vesicle, the primary lipid being selected from the group consisting of C 12 -C 18 fatty alcohols, C 12 -C lg glycol monoesters, C 12 -C 18 gyceryl mono-and diester
  • the invention also relates to a composition
  • a composition comprising a proliferation- inducing amount of a peptide or a salt or derivative thereof encapsulated within a liposome, wherein the peptide is at least 3 amino acids long, has at least 10% sequence identity with the 34 amino acid N-terminal region of hPTH or hPTHrP, and is capable of blocking the inhibition of proliferation or stimulation of differentiation in vitro of cultured human keratinocytes by PTH (1-34), l,25(OH)2D3 or PTHrP (1-34), or in vivo in mouse skin by stimulating skin cell proliferation or accelerating hair cycle progression or stimulating hair growth; wherein said liposome comprises at least two distinct lipids, a primary lipid and a secondary lipid, the primary lipid constituting the greatest proportion, by weight, of any single lipid material forming the bilayers of said vesicle, the primary lipid being selected from the group consisting of C 12 -C 18 fatty alcohols, C 12 -
  • the invention also relates to a composition
  • a composition comprising a proliferation- inhibiting or differentiation-enhancing amount of a peptide or a salt or derivative thereof and an active vitamin D compound, optionally encapsulated within a liposome, wherein the peptide is at least 3 amino acids long, has at least 10% sequence identity with the 34 amino acid N-terminal region of hPTH or hPTHrP, and is capable of inhibiting proliferation or enhancing differentiation in vitro of cultured human keratinocytes, or in vivo in mouse skin by inhibiting skin cell proliferation or hair cycle progression or hair growth.
  • Fig. 1 depicts a bar graph showing the macroscopic effects of topical PTH (7-34) in Novasome on C57BL/6 mice (7 days).
  • Fig. 2 depicts a bar graph showing the effects of PTH (7-34) in
  • Fig. 3 depicts a bar graph showing the topical effects of PTH (1-34) in Novasome on BRDU stained hair follicle cells.
  • Fig. 4 depicts a bar graph showing the effect of 60 days of topical PTH (1-34) on tritiated thymidine incorporation into epidermal DNA in SKH-1 hairless mice.
  • Fig. 5 depicts a bar graph showing the effect of 60 days of topical PTH (7-34) in Novasome on tritiated thymidine incorporation into epidermal DNA in SKH-1 hairless mice.
  • the peptides used in the methods of the invention are all easily synthesized, using recombinant DNA or solid phase peptide synthesis techniques, and some are available commercially as well, or can be derived from commercially available peptides. For example, there is reproduced below a section of the Bach Chem catalog, listing a number of available human, rat, and bovine analogs and fragments. (The Peninsula Laboratory catalog also lists available fragments.)
  • Trp-Leu-Arg-Lys-Lys-Leu-Gln-Asp-Nal-His-Asn-Tyr- ⁇ H 2 (SEQ ID NO:22)
  • Trp-Leu-Arg-Lys-Lys-Leu-Gln-Asp-Nal-His-Asn-Tyr-NH 2 (SEQ ID NO:23)
  • a preferred first step is to choose a peptide which includes a fragment which has at least 10%, and more preferably 50% or greater, sequence identity with an 8 or greater amino acid long fragment within the amino terminal 34 amino acid region of hPTH or hPTHrP.
  • sequence identity refers to a measure of the identity of nucleotide sequences or amino acid sequences. In general, the sequences are aligned so that the highest order match is obtained. "Identity" per se has an art-recognized meaning and can be calculated using published techniques. (See, e.g. : Computational Molecular Biology, Lesk,
  • identity is well known to skilled artisans (Carillo, H. & Lipton, D., SIAM J Applied Math 48:1013 (1988)). Methods commonly employed to determine identity or similarity between two sequences include, but are not limited to, those disclosed in Guide to Huge Computers, Martin J. Bishop, ed., Academic Press, San Diego, 1994, and Carillo, H. & Lipton, D., SIAM J Applied Math 48:1013 (1988). Methods to determine identity and similarity are codified in computer programs.
  • Preferred computer program methods to determine identity and similarity between two sequences include, but are not limited to, GCG program package (Devereux, J., et al, Nucleic Acids Research 12(i):381 (1984)), BLASTP, BLASTN, FASTA (Atschul, S.F., et al, J Molec Biol 275:403 (1990)). Therefore, as used herein, the term "identity" represents a comparison between a test and reference polypeptide. More specifically, reference test polypeptide is defined as any polypeptide that is 10% or more identical to a reference polypeptide. As used herein, the term at least 10% identical to refers to percent identities from 10 to 99.99 relative to the reference polypeptides.
  • Identity at a level of 10% or more is indicative of the fact that, assuming for exemplification purposes a test and reference polynucleotide length of 100 amino acids, that no more than 90% (i.e., 90 out of 100) amino acids in the test polypeptides differ from that of the reference polypeptides.
  • differences may be represented as point mutations randomly distributed over the entire length of the amino acid sequence of the invention or they may be clustered in one or more locations of varying length up to the maximum allowable amino acid difference. Differences are defined as amino acid substitutions, or deletions.
  • the fragment can be modified in any of a variety of standard chemical ways, e.g., the carboxy-terminal amino acid residue can be made into a terminal amide group; the amino-terminal residue can be modified with groups to, e.g., enhance lipophilicity; the peptide can be chemically glycosylated to increase solubility or in vivo half-life; and D-amino acids can be substituted for L-isomers in the peptide.
  • Candidate peptides may be tested for suitability as inhibitors of cell proliferation and enhancers of differentiation using cultured human keratinocytes, as described in U.S. Pat. Nos. 5,527,772, 5,840,690 and 6,066,618. Briefly, those peptides which inhibit proliferation and induce differentiation in cultured keratinocytes are those potentially useful as therapeutic agents in treating disorders, e.g., psoriasis and cancer, where suppression of cell proliferation is desired.
  • Candidate peptides may be tested for suitability as enhancers of cell proliferation using cultured human keratinocytes or in vivo mouse model.
  • Those peptides which block the effect of agonist peptides or l,25(OH)2D3 on cultured keratinocyte proliferation are those potentially useful as therapeutic agents in treating disorders, e.g., wounds, burns, or skin ulcerations, where maintenance or stimulating of cell proliferation is desired.
  • Candidate peptides may be tested for their ability to enhance wound healing by carrying out a skin punch biopsy test, as described in U.S. Pat. Nos. 5,527,772, 5,840,690 and 6,066,618.
  • Candidate peptides may be tested for suitability as stimulators of hair growth using an in vitro hair growth assay, as described in U.S. Pat. Nos. 5,527,772, 5,840,690 and 6,066,618.
  • Those peptides which stimulate hair growth in vitro are those potentially useful for the stimulation of hair growth in vivo, e.g., for the stimulation or maintenance of hair growth during or following chemotherapy or to treat a form of alopecia, e.g., male pattern baldness.
  • in vivo assays may be carried out as described herein and in Schilli, M.B. et al, J. Invest. Dermatol 05:928-932 (1997); Holick, M.F., et al, Proc. Natl. Acad. Sci. P7:8014-8016 (1994); Paus, R. and Cotsarelis,
  • Peptides which block antiproliferative compounds can also be useful in conjunction with chemotherapeutic agents in the treatment of skin cancer; many chemotherapeutic agents are effective only against dividing cells, and the blocking peptides can have the effect of inducing division of otherwise dormant cells, rendering them vulnerable to the chemotherapy. Blocking peptides can also be useful in promoting growth of new cells, e.g., skin cells, in topical skin creams.
  • Differentiation-inducing peptides can be used as immunostimulants, by inducing maturation of monocytes and lymphocytes bearing PTH receptors, while blocking peptides can be used to inhibit lymphocyte maturation, and thus can be used to treat conditions, e.g., autoimmune diseases such as juvenile diabetes, rheumatoid arthritis, and allograft rejection, where mature lymphocytes are a causative agent.
  • the peptides are administered in therapeutically effective amounts to mammals in need of them.
  • the peptides may be administered as part of liposomal preparations described in U.S. Pat. 5,260,065.
  • Such liposomes comprise at least two distinct lipids, a primary lipid and a secondary lipid, the primary lipid constituting the greatest proportion, by weight, of any single lipid material forming the bilayers of said vesicle, the primary lipid being selected from the group consisting of C 12 -C 18 fatty alcohols, C 12 -C 18 glycol monoesters, C 12 -C ⁇ 8 gyceryl mono-and diesters, and mixtures thereof, and the primary lipid further having the property that it will to form a lipid vesicle in the absence of the secondary lipid, and the secondary lipid being present in an amount sufficient to allow formation of the lipid vesicles, the secondary lipid being selected from the group consisting of quaternary dimethyldiacyl amines, poly
  • the preferred primary lipids are C 12 -C 18 fatty alcohols, glyceryl mono- and distearate, glyceryl dilaurate, and glycol stearate. While any of the secondary lipids could be used with any of the primary lipids, preferred combinations include polyoxyethylene 10-20 acyl alcohols or quaternary dimethyldiacyl amines as the secondary lipids to be used in conjunction with the fatty alcohols. Matching chain lengths in terms of carbon content and unsaturations is an important factor to consider for selection of the secondary lipid.
  • acyl alcohols and dimethyldiacyl (specifically distearyl) amines are also useful with the glycol stearate, glyceryl monostearate, glyceryl distearate and the glyceryl dilaurate.
  • the glyceryl distearate and glyceryl dilaurate may also use sodium laurate sarcosinates, as well as other matching sarcosinate salts (all being water soluble), or lauryl sarcosinates as secondary lipids.
  • a sterol such as cholesterol is a particularly useful additive.
  • the addition of cholesterol appears to make the vesicles population more uniform in terms of size and shape. Even cholesterol is not sufficient, in itself, to allow vesicle formation. This is contrast to the materials described in U.S. Pat. No.4,917,951 which only require cholesterol to make vesicles. In certain circumstances, cholesterol will allow these materials which will not otherwise form a lamellar phase to form a lamellar phase but they cannot be formed into vesicles without the addition of the secondary lipid.
  • some of the most preferred secondary lipids e.g., dimethyldistearyl amine, water soluble polyoxyethylene acyl alcohols, and acyl sarcosinate salts, will not form vesicles or lamellar phases either.
  • Example 1 of U.S. Pat. 5,260,065 a variety of materials may be blended in order to make vesicles.
  • Table 1 shows the composition, water uptake level, and oil uptake under hot and cold loading techniques of five different compositions.
  • none of the primary lipids used e.g., glyceryl dilaurate (GDL), glyceryldistearate (GDS), cetyl alcohol (CA), stearyl alcohol (SA), or glycol stearate (GS) will form vesicles or lamellar phase on their own.
  • GDL glyceryl dilaurate
  • GDS glyceryldistearate
  • CA cetyl alcohol
  • SA stearyl alcohol
  • GS glycol stearate
  • the first compound shown in Table 1 is a blend of glyceryl dilaurate, dimethyldicetyl quaternary amine (C16Q), and cholesterol (Choi) in a 1.0:0.05:0.05 molar ratio.
  • the water uptake is 13.5 ml/ml of lipid and the hot load and cold loading values were >7.2 and >2.7 ml of oil/ml of lipid, respectively.
  • the vesicles were made by blending the two lipids and the cholesterol at 70°-75° C. with the aqueous phase at 65° C. According to U.S. Pat.
  • the second compounded tested was a blend of glyceryl distearate, Polyoxyethylene 10 stearyl alcohol (POE10SA), and cholesterol in a 1.0:0.5:0.25 molar ratio.
  • This blended material had a water uptake of 12.5 ml/ml lipid and the oil uptake for either hot and cold loading was >6.5 ml/ml using the same techniques previously described.
  • the third material tested was a blend of cetyl alcohol, polyoxyethylene 10 cetyl alcohol (POE10CA), and cholesterol in a 1:0.2:0.1 molar ratio. Water uptake was 9.5 ml/ml and both hot and cold oil uptake was >4.2 ml/ml lipid.
  • the fourth combination tested was a blend of stearyl alcohol, dimethyldistearyl quaternary amine (C18Q), and cholesterol on a 1:0.2:0.1 ratio. Water uptake was 13.5 ml/ml and oil uptake on both a hot and cold basis was >6.5 ml/ml lipid.
  • the fifth compound tested was a blend of glycol stearate, polyoxyethylene 10 stearyl alcohol, and cholesterol in a 1:0.2:0.1 ratio. Again, the water uptake was approximately 13.5 ml/ml and the oil uptake was >6.5 ml/ml under both hot and cold loading techniques.
  • retinoic acid a water insoluble material in a water immiscible carrier, was used in lieu of the mineral oil of Example 1 in the amorphous central cavity of the paucilamellar lipid vesicles.
  • Retinoic acid has a substantial number of dermatological uses including, potentially, the reduction of facial wrinkles. Table 2
  • Table 2 shows the formulas for two different retinoic acid formulations, one using a cetyl alcohol/polyoxyethylene 10 cetyl alcohol blend and the other using a glycol stearate/polyoxyethylene 10 stearyl alcohol blend as the vesicles formers.
  • Table 2 shows the formulas for two different retinoic acid formulations, one using a cetyl alcohol/polyoxyethylene 10 cetyl alcohol blend and the other using a glycol stearate/polyoxyethylene 10 stearyl alcohol blend as the vesicles formers.
  • both formulas include cholesterol while one uses a mixture petrolatum and paraffin wax as a carrier for the retinoic acid while the other uses a soybean oil carrier.
  • the retinoic acid was dissolved in the carrier at 65°-75° C.
  • the lipids and the cholesterol were then heated and blended to homogeneity and the retinoic acid mixture was added and blended therein.
  • an aqueous phase consisting of the deionized water was then heated to approximately 65° C. and the resulting phases were shear mixed to form the vesicles.
  • the syringe method described in Example 1 could be used, a
  • Example 3 of U.S. Pat. No. 5,260,065 two different formulations for encapsulating anthralin, an antipsoriatic, were tested. Table 3 lists the ingredients used in these formulations. According to the present invention, a peptide agonist or antagonist may be substituted for anthralin.
  • this pefrolatum/paraffin wax mixture appears to be particularly advantageous in that micro-crystals form rather than the macroscopic crystals which normally appear when anthralin cools.
  • the glyceryl distearate, cholesterol and dimethyldistearyl ammonium chloride are blended together at approximately 75° C. until clear and the anthralin solution (forming a water immiscible phase) is then mixed therein.
  • the aqueous phase is formed by heating the deionized water to approximately 65° C.
  • the aqueous phase and the lipid phase are then shear mixed, using a NovaMixTM machine as described in Example 2, to form vesicles.
  • formulation D the cetyl alcohol, polyoxyethylene 10 cetyl alcohol and the cholesterol are blended together at an elevated temperature, the anthralin solution is mixed in, and the aqueous which consists merely of the deionized water is shear mixed using the NovaMixTM machine to form the vesicles.
  • the difference in the procedure is that the non-ionic lipids of formulation D cannot be carried in the aqueous solution as is the ionic sodium lauryl sarcosinate of formulation C.
  • Example 4 of U.S. Pat. 5,260,065 three different materials, Vitamin E acetate, levamisole base, and a butter flavor oil were carried in the central cavity of vesicles of the invention. Table 4 shows the formulas for these vesicles. According to the present invention, a peptide agonist or antagonist may be substituted for vitamin E acetate.
  • Vitamin E 2.2 g
  • formulation E uses glyceryl distearate, polyoxyethylene 10 stearyl alcohol, and cholesterol as the lipophilic phase which are blended at 70° C. to obtain a clear, homogeneous solution.
  • Vitamin E acetate was dissolved therein and the mixture was hydrated with 65° C. water using the NovaMixTM machine as described in Example 2.
  • formulation F used a levamisole base (a sheep dip) in soybean oil at 75° C. to form the water immiscible phase.
  • the glycol stearate, polyoxyethylene stearyl alcohol and cholesterol were heated together at 75° C. to obtain a clear, homogeneous solution and the levamisole/soybean oil mixture was blended therewith.
  • the deionized water was heated to approximately 65° C. and used as a hydrating solution for the lipids, again using the previously described NovaMixTM machine.
  • formulation H uses the paraffin wax/petrolatum carrier for the retinoic acid, with the retinoic acid being dissolved in the carrier at approximately 65°-75° C. According to U.S. Pat. 5,260,065, formulation H uses the paraffin wax/petrolatum carrier for the retinoic acid, with the retinoic acid being dissolved in the carrier at approximately 65°-75° C. According to U.S. Pat.
  • the lipophilic phase is formed of glyceryl distearate, cholesterol, and the dimethyl distearyl ammonium chloride.
  • the carrier containing the retinoic acid is blended into the lipophilic phase and is hydrated with the deionized water using the NovaMixTM machine as described in Example 2.
  • formulations I and J use the soybean oil carrier and the same materials except for the secondary lipid.
  • the secondary lipid which forms part of the initial lipophilic phase, is dimethyl dicetyl ammonium chloride while in formulation J, the secondary lipid, which is incorporated into the aqueous phase, is sodium oleate.
  • the retinoic acid is dissolved in the soybean oil at elevated temperatures, the soybean oil is blended into the lipophilic phase, and the combined phase is then hydrated using the aqueous phase.
  • formulation J forms anionic vesicles while formulation I forms cationic vesicles.
  • a pharmacologically inert topical carrier such as one comprising a gel, an ointment or a cream, including such carriers as water, glycerol, alcohol, propylene glycol, fatty alcohol, triglycerides, fatty acid ester or mineral oils.
  • liquid petrolatum isopropylpalmitate, polyethylene glycol ethanol 95%, polyoxyethylene monolaurate 5% in water, sodium lauryl sulfate 5% in water, and the like.
  • Materials such as antioxidants, humectants, viscosity stabilizers and the like may be added, if necessary.
  • the peptides can be provided in the form of pharmaceutically acceptable salts.
  • preferred salts are those of therapeutically acceptable organic acids, e.g., acetic, lactic, maleic, citric, malic, ascorbic, succinic, benzoic, salicylic, methanesulfonic, toluenesulfonic, or pamoic acid, as well as polymeric acids such as tannic acid or carboxymethyl cellulose, and salts with inorganic acids such as hydrohalic acids, e.g, hydrochloric acid, sulfuric acid, or phsophoric acid.
  • organic acids e.g., acetic, lactic, maleic, citric, malic, ascorbic, succinic, benzoic, salicylic, methanesulfonic, toluenesulfonic, or pamoic acid
  • polymeric acids such as tannic acid or carboxymethyl cellulose
  • salts with inorganic acids such as hydrohalic acids,
  • Dosage will be dependent upon the age, health, and weight of the recipient; kind of concurrent treatment, if any; frequency of treatment; and the nature of the effect desired. Generally, daily dosage will be from about 0.0001 micrograms/kg to 100 micrograms/kg, preferably 0.001 to 10.0 micrograms/kg.
  • the topical dosage will be from about 0.01 micrograms/cm 2 to 100 micrograms/cm 2 , preferably 0.1 to 10 micrograms/cm 2 .
  • the liposomal formulations may be applied by one or more applications per day.
  • the invention also relates to compositions comprising a proliferation- inhibiting or differentiation-enhancing amount of a peptide or a salt or derivative thereof, an active vitamin D compound and a pharmaceutical carrier, wherein the peptide is at least 3 amino acids long, has at least 10% sequence identity with the 34 amino acid N-terminal region of hPTH or hPTHrP, and is capable of inhibiting proliferation or enhancing differentiation in vitro of cultured human keratinocytes, or in vivo in mouse skin by inhibiting skin cell proliferation or hair cycle progression or hair growth.
  • active vitamin D compounds are known which can be used in the practice of the present invention. See U.S. Patent Nos.
  • a preferred active vitamin D compound is calcipotriene.
  • any conventional liposome may be used including the liposomes described in U.S. Pat. 4,235,871, 4,241,046, 4,247,411, 4,356,167, 4,377,567, 4,544,545, 4,551,288, 4,610,868, 4,731,210, 4,744,989, 4,772,471, 4,897,308, 4,917,951, 5,021,200, 5,032,457, and 5,260,065.
  • the invention relates as well to a method of inhibiting proliferation or enhancing differentiation of a skin or hair cell of a mammal, comprising administering to the mammal in need thereof a proliferation-inhibiting or differentiation-enhancing amount of a peptide or a salt or derivative thereof and an active vitamin D compound, wherein the peptide is at least 3 amino acids long, has at least 10% sequence identity with the 34 amino acid N- terminal region of hPTH or hPTHrP, and is capable of inhibiting proliferation or enhancing differentiation in vitro of cultured human keratinocytes, or in vivo in mouse skin by inhibiting skin cell proliferation or hair cycle progression or hair cell growth.
  • the peptide and the active vitamin D compound may be administered as part of single or separate pharmaceutical compositions. Either one or both of the peptide and active vitamin D compound may be administered topically or parenterally. In a preferred embodiment, the peptide is administered first followed by the active vitamin D compound.
  • PTH (1-34) and PTH (7-34) was formulated in Novasome A (obtained from IGI, Inc., Buena, NJ).
  • the formulation included dissolving the PTH analog in doubly distilled water at a concentration of 1 mg/100 ⁇ l. This solution was mixed with 3 ml of Novasome A.
  • Example 2 Effect of Liposome Encapsulated Peptides on Skin and Hair Growth in Mice
  • C57 BL/6 were purchased from Jackson Labs and were in their telogen state of hair development. The animals were depilated as previously described (1). Groups of six animals received daily a topical application of either 10 ⁇ g of PTH (1-34) in 30 ⁇ l of Novasome or their depilatated backs, 10 ⁇ g of PTH
  • SKH-1 hairless mice received topically daily for 60 days either 10 ⁇ g of PTH (1-34) formulated in Novasome A, PTH (7-34) formulated in Novasome A, or Novasome A without any PTH analog to serve as the control group.
  • the animals received 3 H-tritiated thymidine for the evaluation of epidermal proliferation as previously described.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Chemical & Material Sciences (AREA)
  • Epidemiology (AREA)
  • Dermatology (AREA)
  • Endocrinology (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Biophysics (AREA)
  • Birds (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Toxicology (AREA)
  • Biochemistry (AREA)
  • Dispersion Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Immunology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicinal Preparation (AREA)
  • Cosmetics (AREA)
EP01946566A 2000-06-22 2001-06-20 Lokale applikation von peptiden zur regulation der zellproliferation und zelldifferenzierung Withdrawn EP1349564A2 (de)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US21324700P 2000-06-22 2000-06-22
US213247P 2000-06-22
PCT/US2001/019650 WO2001098348A2 (en) 2000-06-22 2001-06-20 Regulation of cellproliferation and differentiation using topically applied peptides

Publications (1)

Publication Number Publication Date
EP1349564A2 true EP1349564A2 (de) 2003-10-08

Family

ID=22794320

Family Applications (1)

Application Number Title Priority Date Filing Date
EP01946566A Withdrawn EP1349564A2 (de) 2000-06-22 2001-06-20 Lokale applikation von peptiden zur regulation der zellproliferation und zelldifferenzierung

Country Status (4)

Country Link
EP (1) EP1349564A2 (de)
AU (1) AU2001268594A1 (de)
CA (1) CA2413860A1 (de)
WO (1) WO2001098348A2 (de)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2003207512B2 (en) 2002-01-10 2008-06-12 Osteotrophin Llc Treatment of bone disorders with skeletal anabolic drugs
CN1822853A (zh) * 2003-07-15 2006-08-23 加拿大国家研究委员会 用于治疗特征在于过度增殖皮肤细胞的疾病的人类甲状旁腺激素环状类似物
FR2857597B1 (fr) 2003-07-18 2008-01-25 Inst Europeen Biolog Cellulair Utilisation de conjugues peptidiques pour la preparation de compositions destinees au traitement preventif et curatif de l'alopecie.
FR2857667B1 (fr) 2003-07-18 2007-07-13 Inst Europ Biolog Cellulaire Nouveaux conjugues peptidiques utiles dans le traitement preventif et curatif de l'alopecie
CN109182250A (zh) * 2018-09-30 2019-01-11 山东省立医院 一种小鼠耳蜗毛细胞培养方法

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4692433A (en) * 1983-10-12 1987-09-08 The Regents Of The University Of California Method and composition for regulating serum calcium levels of mammals
US5527772A (en) * 1987-10-20 1996-06-18 Holick; Michael F. Regulation of cell proliferation and differentiation using peptides
US5260065A (en) * 1991-09-17 1993-11-09 Micro Vesicular Systems, Inc. Blended lipid vesicles
DE69429337T3 (de) * 1993-04-02 2012-08-30 Anticancer Inc. Verfahren zur verabreichung von förderlichen zusammensetzungen auf die haarfollikel
US5744128A (en) * 1995-05-03 1998-04-28 Holick; Michael F. Use of emu oil for stimulating skin and hair growth
US6300341B1 (en) * 1998-09-30 2001-10-09 The Procter & Gamble Co. 2-substituted heterocyclic sulfonamides

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0198348A2 *

Also Published As

Publication number Publication date
AU2001268594A1 (en) 2002-01-02
WO2001098348A2 (en) 2001-12-27
WO2001098348A3 (en) 2003-07-31
CA2413860A1 (en) 2001-12-27

Similar Documents

Publication Publication Date Title
US6066618A (en) Regulation of cell proliferation and differentiation using peptides
US20070128265A1 (en) Regulation of cell proliferation and differentiation using topically applied peptides
EP1509243B1 (de) Topische arzneimittelabgabe mit phosphatidylcholin
US5958384A (en) Use of emu oil for stimulating skin and hair growth
US5650166A (en) Moisturizing composition for the simultaneous treatment of the surface layers and deep layers of the skin, and use thereof
US20070224150A1 (en) Growth factor for hair and skin treatment
US20070081963A1 (en) Composition for improving skin conditions comprising human growth hormone as an active ingredient
US20080020993A1 (en) Regulation of cell proliferation and differentiation using topically applied nucleic acid molecules
US5252559A (en) His-Gly-Gly peptide and derivatives thereof for hair growth
US5068315A (en) Composition for the regulation of hair growth
EP1349564A2 (de) Lokale applikation von peptiden zur regulation der zellproliferation und zelldifferenzierung
EP1222913B1 (de) Verfahren zur Enthaarung
EP1513547A2 (de) Verwendung von parathormone-peptid-analoga zur behandlung von scheidenatrophie
JP2802084B2 (ja) ペプチドを用いた細胞の増殖と分化の調節
CN1070369C (zh) 毛发生长的改变
Hintz et al. Synthetic hGH causes both increased somatomedin levels and insulin resistance in humans
Stebbing et al. Efficacy and pharmacokinetics of recombinant DNA derived human growth hormone in rats and monkeys
Bierich et al. Stunted growth due to peripheral resistance to somatomedin, a new syndrom
Sobel et al. Cell mediated immunity to pituitary extracts in hypopituitarism
Aubert et al. Ontogeny of hypothalamic GnRH and pituitary GnRH recepotors in fetal rats
WO1992004039A1 (en) Regulation of cell proliferation and differentiation using peptides
JPH09500090A (ja) 毛髪成長を促進する方法及びルトゾミア・タンパク質を含む毛髪活性化組成物

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20021227

AK Designated contracting states

Kind code of ref document: A2

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE TR

AX Request for extension of the european patent

Extension state: AL LT LV MK RO SI

REG Reference to a national code

Ref country code: HK

Ref legal event code: DE

Ref document number: 1060507

Country of ref document: HK

REG Reference to a national code

Ref country code: HK

Ref legal event code: WD

Ref document number: 1060507

Country of ref document: HK

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20120103