U.S. patent application number 12/175994 was filed with the patent office on 2009-01-22 for cosmetic or dermatological preparations comprising oligopeptides for lightening the skin of age marks and/or for preventing tanning of the skin, in particular tanning of the skin caused by uv radiation.
This patent application is currently assigned to BEIERSDORF AG. Invention is credited to Vincent J. Hearing, Heiner Max, Uwe SCHONROCK.
Application Number | 20090022674 12/175994 |
Document ID | / |
Family ID | 22455656 |
Filed Date | 2009-01-22 |
United States Patent
Application |
20090022674 |
Kind Code |
A1 |
SCHONROCK; Uwe ; et
al. |
January 22, 2009 |
COSMETIC OR DERMATOLOGICAL PREPARATIONS COMPRISING OLIGOPEPTIDES
FOR LIGHTENING THE SKIN OF AGE MARKS AND/OR FOR PREVENTING TANNING
OF THE SKIN, IN PARTICULAR TANNING OF THE SKIN CAUSED BY UV
RADIATION
Abstract
Cosmetic or pharmaceutical preparations which are distinguished
by an effective content of one or more monomeric or homo- or
heterodimeric or homo- or heterotrimeric or homo- or
heterotetrameric oligopeptides, where these oligopeptides are based
on a Structure Val-Val-Arg-Pro SEQ ID NO: 1 as homo- or
heteromonomer.
Inventors: |
SCHONROCK; Uwe; (Nahe,
DE) ; Max; Heiner; (Hamburg, DE) ; Hearing;
Vincent J.; (Clarksburg, MD) |
Correspondence
Address: |
GREENBLUM & BERNSTEIN, P.L.C.
1950 ROLAND CLARKE PLACE
RESTON
VA
20191
US
|
Assignee: |
BEIERSDORF AG
Hamburg
DE
|
Family ID: |
22455656 |
Appl. No.: |
12/175994 |
Filed: |
July 18, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
09132799 |
Aug 13, 1998 |
|
|
|
12175994 |
|
|
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Current U.S.
Class: |
424/59 |
Current CPC
Class: |
A61Q 19/02 20130101;
C07K 7/06 20130101; A61K 8/64 20130101; A61Q 17/04 20130101; C07K
5/101 20130101 |
Class at
Publication: |
424/59 |
International
Class: |
A61K 8/64 20060101
A61K008/64; A61Q 17/04 20060101 A61Q017/04 |
Claims
1. A cosmetic or dermatological topical water-in-oil preparation
for cosmetic and topical dermatological lightening of the skin or
preventing tanning of the skin caused by UV radiation which
comprises of one or more monomeric oligopeptides selected from the
group consisting of: ##STR00012## wherein: R represents a branched
or unbranched, saturated or unsaturated alkyl radical having
C.sub.1-C.sub.30 carbon atoms, R' and R'' independently of one
another may be selected from the group consisting of hydrogen and
branched or unbranched, saturated of unsaturated alkyl radical
having C.sub.1-C.sub.30 carbon atoms, one or more cosmetically or
dermatologically acceptable active ingredients, auxiliaries and/or
additives; and a cosmetically or dermatologically acceptable
carrier.
2. The topical water-in-oil preparation of claim 1, wherein the
oligopeptides(s) is/are present in cosmetic or dermatological
topical preparations in concentration of 0.000001-10% by weight,
based on the total weight of the preparations.
3. The topical water-in-oil preparation of claim 2, wherein the
oligopeptides(s) is/are present in the cosmetic or dermatological
topical preparations in concentrations of 0.0001-1% by weight based
on the total weight of the preparations.
4. The topical water-in-oil preparation of claim 3, wherein the
oligopeptides(s) is/are present in the cosmetic or dermatological
topical preparations in concentrations of 0.0001-0.1% by weight
based on the total weight of the preparations.
5. The cosmetic or dermatological topical water-in-oil preparation
of claim 1 wherein the oligopeptide is ##STR00013##
6. The cosmetic or dermatological topical water-in-oil preparation
of claim 1 wherein the oligopeptide is ##STR00014##
7. The cosmetic or dermatological topical water-in-oil preparation
of claim 1 wherein the oligopeptide is ##STR00015##
8. The cosmetic or dermatological topical water-in-oil preparation
of claim 1 wherein the oligopeptide is ##STR00016##
9. The cosmetic or dermatological topical water-in-oil preparation
of claim 6 wherein R' and R'' is hydrogen.
10. The cosmetic or dermatological topical water-in-oil preparation
of claim 7 wherein R is methyl.
11. The cosmetic or dermatological topical water-in-oil preparation
of claim 7 wherein R is an n-C.sub.15 or n-C.sub.17 alkyl
radical.
12. The cosmetic of dermatological topical water-in-oil preparation
of claim 8 wherein R is methyl, R' is hydrogen and R'' is hydrogen.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of application Ser. No.
09/132,799, filed Aug. 13, 1998, which is incorporated by reference
herein in its entirety.
[0002] The present invention relates to cosmetic or dermatological
preparations comprising oligopeptides, for using active ingredients
known per se, for cosmetic and topical dermatological lightening of
the skin or for preventing tanning of the skin, in particular
tanning of the skin caused by TV radiation.
[0003] In a preferred embodiment, the present invention relates to
cosmetic and dermatological preparations for the prophylaxis and
treatment of cosmetic or dermatological changes in the skin such
as, for example, undesired pigmentation, for example local
hyperpigmentations and malpigmentations (for example lentigo,
freckles), but also for the purely cosmetic lightening of large
areas of skin whose pigmentation is quite adequate for the
individual skin type.
[0004] The factors which are responsible for skin pigmentation are
the melanocytes which are found in the lowest layer of the
epidermis, the stratum basale, next to the basal cells, as
pigment-forming cells which occur either singly or more or less in
clusters, depending on the skin type Melanocytes contain
melanosomes as characteristic cell organelles, and these form
melanin at higher rates when stimulated by UV radiation. The
melanin is transported into the keratinocytes and leads to a more
or less pronounced tan or brown skin colour.
[0005] Melanin is the end product of an oxidative process in which
tyrosine is converted with the aid of the enzyme tyrosinase via
3,4-dihydroxyphenylalanine (dopa), dopa-quinone, leucodopachrome,
dopachrome, 5,6-dihydroxyindole and indole-5,6-quinone to give,
finally, melanin.
[0006] Hyperpigmentation problems of the skin have a multiplicity
of causes and/or accompanying symptoms of a large number of
biological processes, for example UV radiation (for example,
freckles, ephelides), genetic predisposition, malpigmentation of
the skin in the course of wound healing and/or wound scarring or
skin ageing (for example lentigines seniles).
[0007] Active ingredients and preparations which counteract skin
pigmentation are known. Products which are essentially used in
practice are based on hydroquinone but, on the one hand, only
become effective after several weeks' use and, on the other hand,
unduly long use of such products is unacceptable for toxicological
reasons. Inhibition of tyrosinase by substances such as kojic acid,
ascorbic acid and azelaic acid and their derivatives is also well
known, but has cosmetic and dermatological disadvantages.
[0008] It was an object of the present invention to remedy these
shortcomings.
[0009] The publication WO-97/00892 describes the depigmenting
action of the signal proteins of agoutis (rodent family of the
Dasyproctidae, occurrence: approx. 20 species in the woodlands of
Central America).
[0010] Surprisingly, the object is achieved by cosmetic or
pharmaceutical preparations which are distinguished by an effective
content of one or more monomeric or homo- or heterodimeric or homo-
or heterotrimeric or homo- or heterotetrameric oligopeptides,
[0011] (1) Where these oligopeptides are based on a structure
Val-Val-Arg-Pro (or VVRP) SEQ ID NO: 1 as homo- or heteromonomer,
[0012] (2) where, in the sequence Val-Val-Arg-Pro, in each case one
valine may be replaced by an amino acid selected from the group
consisting of glycine, alanine, leucine, isoleucine, serine,
threonine and methionine, preferably alanine, while the remainder
of the sequence is retained, [0013] (3) or where structures as
mentioned above are present in the monomeric or homo- or
heterodimeric or homo- or heterotrimeric or homo- or
heterotetrameric oligopeptides, with the difference that the
C-terminus and/or the N-terminus is complemented by up to 5 amino
acids, as shown by the structure
##STR00001##
[0013] where .psi. and .OMEGA. independently of one another may
represent amino acid sequences of 0 up to 5 amino acids, [0014] (4)
or where amino acid sequences shown under items (1) and (2) occur
as single or repetitive structural motifs in monomeric or
homo-/hetero-di-, -tri- or -tetrameric oligopeptides or proteins
with a molecular weight of between approx 0.5 and 100 kdalton,
[0015] (5) or where the monomeric or homo-/hetero-di-, -tri-, or
-tetrameric oligopeptides or proteins are acylated on the
N-terminus, as shown by the structure
##STR00002##
[0015] where .psi. and .OMEGA. independently of one another may
represent amino acid sequences of 0 up to 5 amino acids and R
represents a branched or unbranched, saturated or unsaturated
hydrocarbon radical, in particular an alkyl radical having 1 to 30
carbon atoms, [0016] (6) or where the monomeric or
homo-/hetero-di-, -tri-, or -tetrameric oligopeptides or proteins
are amidated on the C-terminus, as shown by the structure
##STR00003##
[0016] where .psi. and .OMEGA. independently of one another may
represent amino acid sequences of 0 up to 5 amino acids and where
R' and R'' independently of one another may be selected from the
group consisting of hydrogen and the branched or unbranched,
saturated or unsaturated hydrocarbon radicals, in particular the
alkyl radical having 1 to 30 carbon atoms, [0017] (6) or where the
monomeric or homo-/hetero-di-, -tri-, or -tetrameric oligopeptides
or proteins are amidated on the C-terminus and acylated on the
N-terminus, as shown by the structure
##STR00004##
[0017] where .psi. and .OMEGA. independently of one another may
represent amino acid sequences of 0 up to 5 amino acids, R
represents a branched or unbranched saturated or unsaturated
hydrocarbon radical, in particular an alkyl radical having 1 to 30
carbon atoms, and where R' and R'' independently of one another may
be selected from the group consisting of hydrogen and the branched
or unbranched, saturated or unsaturated hydrocarbon radicals, in
particular the alkyl radical having 1 to 30 carbon atoms, [0018]
(8) or where amino acid sequences occur in monomeric or
homo-/hetero-di-, -tri- or -tetrameric oligopeptides or proteins as
single or repetitive structural motifs, as shown by the
structures
##STR00005##
[0018] where .psi. may represent a single bond or an amino acid
sequence of up to 5 amino acids, .OMEGA. may represent an amino
acid sequence of up to 15 amino acids, n represents a number from 1
to 25, R represents a branched or unbranched saturated or
unsaturated hydrocarbon radical, in particular an alkyl radical of
1 to 30 carbon atoms, and where R' and R'' independently of one
another may be selected from the group consisting of hydrogen and
the branched or unbranched, saturated or unsaturated hydrocarbon
radicals, in particular the alkyl radical having 1 to 30 carbon
atoms.
[0019] Also considered to be an advantageous embodiment of the
present invention is the use of one or more monomeric or homo- or
heterodimeric or homo- or heterotrimeric or homo- or
heterotetrameric oligopeptides, [0020] (1) where these
oligopeptides are based on a structure Val-Val-Arg-Pro (or VVRP)
SEQ ID NO: 1 as homo- or neteromonomer, [0021] (2) where, in the
sequence Val-Val-Arg-Pro, in each case one valine may be replaced
by an amino acid selected from the group consisting of glycine,
alanine, leucine, isoleucine, serine, threonine and methionine,
preferably alanine, while the remainder of the sequence is
retained, [0022] (3) or where structures as mentioned above are
present in the monomeric or homo- or heterodimeric or homo- or
heterotrimeric or homo- or heterotetrameric oligopeptides, with the
difference that the C-terminus and/or the N-terminus is
complemented by up to 5 amino acids, as shown by the structure
##STR00006##
[0022] where .psi. and .OMEGA. independently of one another may
represent amino acid sequences of 0 up to 5 amino acids, [0023] (4)
or where amino acid sequences shown under items (1) and (2) occur
as single or repetitive structural motifs in monomeric or
homo-/hetero-di-, -tri- or -tetrameric oligopeptides or proteins
with a molecular weight of between approx. 0.5 and 100 kdalton,
[0024] (5) or where the monomeric or homo-/hetero-di-, -tri-, or
-tetrameric oligopeptides or proteins are acylated on the
N-terminus, as shown by the structure
##STR00007##
[0024] where .psi. and .OMEGA. independently of one another may
represent amino acid sequences of 0 up to 5 amino acids and R
represents a branched or unbranched saturated or unsaturated
hydrocarbon radical, in particular an alkyl radical having 1 to 30
carbon atoms, [0025] (6) or where the monomeric or
homo-/hetero-di-, -tri-, or -tetrameric oligopeptides or proteins
are amidated on the C-terminus, as shown by the structure
##STR00008##
[0025] where .psi. and .OMEGA. independently of one another may
represent amino acid sequences of 0 up to 5 amino acids and R
represents a branched or unbranched saturated or unsaturated
hydrocarbon radical, in particular an alkyl radical having 1 to 30
carbon atoms, [0026] (6) or where the monomeric or
homo-/hetero-di-, -tri-, or -tetrameric oligopeptides or proteins
are amidated on the C-terminus, as shown by the structure
##STR00009##
[0026] where .psi. and .OMEGA. independently of one another may
represent amino acid sequences of 0 up to 5 amino acids, R
represents a branched or unbranched saturated or unsaturated
hydrocarbon radical, in particular an alkyl radical having 1 to 30
carbon atoms, and where R' and R'' independently of one another may
be selected from the group consisting of hydrogen and the branched
or unbranched, saturated or unsaturated hydrocarbon radicals, in
particular the alkyl radical having 1 to 30 carbon atoms, [0027]
(8) or where amino acid sequences occur in monomeric or
homo-/hetero-di-, -tri- or -tetrameric oligopeptides or proteins as
single or repetitive structural motifs, as shown by the
structures
##STR00010##
[0027] where .psi. may represent a single bond or an amino acid
sequence of up to 5 amino acids, .OMEGA. may represent an amino
acid sequence of up to 15 amino acids, n represents a number from 1
to 25, R represents a branched or unbranched saturated or
unsaturated hydrocarbon radical, in particular an alkyl radical of
1 to 30 carbon atoms, and where R' and R'' independently of one
another may be selected from the group consisting of hydrogen and
the branched or unbranched, saturated or unsaturated hydrocarbon
radicals, in particular the alkyl radical having 1 to 30 carbon
atoms.
[0028] The amino acid sequences which optionally complement the
peptide sequence on the C- or N-terminus are preferably constituted
of proteinogenic amino acids.
[0029] The proteinogenic amino acids and their corresponding three-
and single-letter codes are shown in Table 1.
TABLE-US-00001 TABLE Three- and single-letter codes for amino acids
Amino acid L-alanine Ala A L-valine Val V L-leucine Leu L
L-isoleucine Ile I L-proline Pro P L-tryptophan Typ L-phenylalanine
Phe F L-methionine Met M glycine Gly G L-serine Ser S L-tyrosine
Tyr Y L-threonine Thr T L-cysteine Cys C L-asparagine Asn N
L-glutamine Gln Q L-aspartic acid Asp D L-glutamic acid Glu E
L-lysine Lys K L-arginine Arg R L-histidine His H
[0030] L-Selenocysteine may replace L-cysteine, and
L-seleno-methionine L-methionine. In addition, individual, several
or else all positions may also be replaced by the corresponding
stereoisomers in the D-configuration.
[0031] Examples of especially advantageous oligopeptides according
to the invention are Val-Val-Arg-Pro SEQ ID NO: 1,
Val-Val-Arg-Pro-Pro SEQ ID NO: 2, and Val-Val-Arg-Pro-Pro-Pro SEQ
ID NO: 3.
[0032] Particularly advantageous according to the invention are the
products of the oligopeptides Val-Val-Arg-Pro SEQ ID NO: 1,
Val-Val-Arg-Pro-Pro SEQ ID NO: 2, and Val-Val-Arg-Pro-Pro-Pro SEQ
ID NO: 3 which are acylated on the N-terminus and/or amidated on
the C-terminus.
[0033] Preferred acylated oligopeptides are those which are
acylated on the N-terminus with unbranched alkanoyl groups.
[0034] Particularly preferred are
##STR00011##
[0035] R representing the stearoyl or the palmitoyl radical.
[0036] It may be preferable to synthesize the peptides of the
present invention using recombinant DNA methods. Alternatively, it
may be preferable to synthesize the peptides of the present
invention using the well-known chain elongation techniques such as
solid-phase synthesis, as on a Merrifield resin or the like.
[0037] To synthesize a peptide using recombinant DNA, one typically
synthesizes a double-stranded DNA chain which encodes the desired
amino acid sequence. The degeneracy of the genetic code permits a
wide variety of codon combinations to be used to form the DNA chain
that encodes the product peptide. Certain particular codons are
more efficient for peptide expression in certain types of organisms
and the selection of codons preferably is made according to those
codons which are most efficient for expression in the type of
organism which is to serve as the host for the recombinant vector.
However, any correct set of codons should encode the desired
product, even if slightly less efficiently. Codon selection may
also depend upon vector construction considerations, for example,
it may be necessary to avoid creating a particular restriction site
in the DNA chain if, subsequent to insertion of the synthetic DNA
chain, the vector is to be manipulated using a restriction enzyme
that cleaves at such a site. Also, it is necessary to avoid placing
restriction sites in the DNA chain if the host organism which is to
be transformed with the recombinant vector containing the DNA chain
is known to produce a restriction enzyme that would cleave at such
a site within the DNA chain.
[0038] In addition to the encoding sequences, the DNA chain that is
synthesized may contain additional sequences, depending upon vector
construction considerations. Typically, a DNA chain is synthesized
with linkers at its ends to facilitate insertion into restriction
sites within a cloning vector. The DNA chain may be constructed so
as to encode the desired sequence as a portion of a fusion peptide
and if so, it may contain terminal sequences that encode amino acid
residue sequences that serve as proteolytic processing sites,
whereby the desired peptide may be proteolytically cleaved from the
remainder of the fusion peptide. The terminal portions of the
synthetic DNA chain may also contain appropriate start and stop
signals.
[0039] To assemble the desired DNA chain, oligonucleotides are
constructed by conventional methods such as procedures described in
T. Maniatis et al. COLD SPRING HARBOR LABORATORY MANUAL, Cold
Spring Harbor, N.Y. (1982) (hereinafter CSH). Sense and antisense
oligonucleotide chains up to about 70 nucleotide residues long are
synthesized preferably on automated synthesizers such as the
Applied Biosystems in model 380B DNA synthesizer. The
oligonucleotide chains are constructed so that the sense and
antisense oligonucleotides associate with each other through
hydrogen bonding between complementary base pairs and thereby form
double-stranded chains. These oligonucleotides are then ligated to
the vector.
[0040] The cloning vector that is to be recombined to incorporate
the DNA chain is selected appropriate to its viability and
expression in a host organism or cell line, and the manner of
insertion of the DNA chain depends upon factors particular to the
host. For example, if the DNA chain is to be inserted into a vector
for insertion into a prokaryotic cell, such as E. coli, the DNA
chain will be inserted 3' of a promoter sequencer a Shine-Delgarno
sequence (or ribosome binding site) that is within a 5'
non-translated portion and an ATG start codon. The ATG start codon
is appropriately spaced from the Shine-Delargo sequence, and the
encoding sequence is placed in correct reading frame with the ATG
start codon. The cloning vector also provides a 3' non-translated
region and a transcription termination site. A translation
termination site could be provided by either the synthetic DNA or
the cloning vector. For insertion into a eukaryotic cell, such as a
yeast cell or a cell line obtained from a higher animal, the
encoding oligonucleotide sequence is appropriately spaced from a
capping site and in correct reading frame with an ATG start signal.
The cloning vector also provides a 3' non-translated region and a
polyadenylation site. A translation termination site could be
provided by either the synthetic DNA or the vector.
[0041] Derivatives of prokaryotic vectors, such as pBR322, pMB9,
Col El, pCRI, RP4 and lambda-phage, are available for inserting a
DNA chain of the length necessary to encode peptides of interest
with substantial assurance of at least some expression of the
encoded peptide. Typically, such vectors are constructed or
modified to have a unique restriction site(s) appropriately
positioned relative to a promoter, such as the lac promoter. The
DNA chain may be inserted with appropriate linkers into such a
restriction site, with substantial assurance of production of
peptide in a prokaryotic cell line transformed with the recombinant
vector. To assure the proper reading frame, linkers of various
lengths may be provided at the ends of the peptide-encoding
sequence. Cassettes, which include sequences, such as the 5' region
of the lac Z gene (including the operator, promoter, transcription
start site, Shine Delgarno sequence and translation initiation
signal), the regulatory region from the tryptophan gene (trp
operator, promoter, ribosome binding site and translation
initiator) and a fusion gene containing these two promoters, called
the trp-lac or commonly called the Tac promoter, are available into
which a synthetic DNA chain may be conveniently inserted.
[0042] Similarly, eukaryotic transformation vectors, such as the
cloned bovine papilloma virus genome, the cloned genomes of the
murine retroviruses, and eukaryotic cassettes, such as the pSV-2
gpt system (described by Mulligan and Berg, NATURE, Vol. 277, pp.
108-114, 1979), the Okayama-Berg cloning system (MOL. CELL BIOL.,
Vol. 2, pp. 161-170, 1982) and the expression cloning vector
recently described by Genetics Institute (SCIENCE, Vol. 228, pp.
810-815, 1985). These provide substantial assurance of at least
some expression of the peptide in the transformed eukaryotic cell
line.
[0043] Another way to produce desired peptides is to produce the
peptide initially as a segment of a gene-encoded fusion peptide. In
such a case, the DNA chain is constructed so that the expressed
peptide has enzymatic processing sites flanking the peptide
sequence or, more commonly, processing site at one side of the
desired peptide. A peptide-encoding DNA chain may be inserted for
example, into the beta-galactosidase gene for insertion into E.
coli, in which case, the expressed fusion peptide is subsequently
cleaved with appropriate proteolytic enzymes to release the peptide
from beta-galactosidase peptide sequences.
[0044] Alternatively, the peptides can be synthesized by suitable
chain elongation or coupling-type methods, such as by exclusively
solid-phase techniques, by partial solid-phase techniques, by
fragment condensation or by classical solution couplings. The
techniques of exclusively solid-phase synthesis are set forth in
the textbook "Solid-Phase Peptide Synthesis" Stewart & Young,
Pierce Chemical Company, Rockford, Ill., 1984 and are exemplified
by the disclosure of U.S. Pat. No. 4,105,603, issued Aug. 8, 1978,
incorporated herein by reference. The fragment condensation method
of synthesis is exemplified in U.S. Pat. No. 3,972,859 issued Aug.
3, 1976, incorporated herein by reference. Other available
syntheses are exemplified by U.S. Pat. No. 3,842,067, issued Oct.
15, 1974 and U.S. Pat. No. 3,862,925 issued Jan. 28, 1975, both
incorporated herein by reference.
[0045] Common to coupling-type syntheses is the protection of the
labile side chain groups of the various amino acid moieties with
suitable protecting groups which will prevent a chemical reaction
from occurring at that site until the group is ultimately removed.
Usually also common is the protection of an alpha-amino group by an
amino-terminus blocking group on an amino acid or a fragment while
that entity reacts at the carboxyl group, followed by the selective
removal of the amino-terminus blocking group to allow subsequent
reaction to take place at that location. Accordingly, it is common
that, as a step in the synthesis, an intermediate compound is
produced which includes each of the amino acid residues located in
its desired sequence in the peptide chain with side-chain
protecting groups or terminus blocking group linked to the
appropriate residues.
[0046] In selecting a particular side chain protecting group to be
used in the synthesis of the peptides, the following rules are
followed: (a) the protecting group should be stable to the reagent
and under the reaction conditions selected for removing the
.alpha.-amino protecting group at each step of the synthesis, (b)
the protecting group should retain its protecting properties and
not be split off under coupling conditions, and (c) the side chain
protecting group should be removable, upon the completion of the
synthesis containing the desired amino acid sequence, under
reaction conditions that will not alter the peptide chain.
[0047] The peptides are preferably prepared using solid phase
synthesis, such as that described by Merrifield, J. AM. CHEM. SOC.,
Vol. 85, pp. 2149 (1963), although other equivalent chemical
syntheses known in the art can also be used as previously
mentioned. Solid-phase synthesis is commenced from the C-terminal
end of the peptide by coupling a protected a-amino acid to a
suitable resin. Such a starting material can be prepared by
attaching a-amino-protected Val by an ester linkage to a
chloromethylated resin or a hydroxymethyl resin, or by an amide
bond to a butylated hydroxy anisole (BHA) resin or MBHA resin. The
preparation of the hydroxymethyl resin is described by Bodansky et
al., CHEM. IND (London) Vol. 38, pp. 1597-98 (1966).
Chloromethylated resins are commercially available from Bio Rad
Laboratories, Richmond, Calif. and from Lab. Systems, Inc. The
preparation of such a resin is described by Stewart et al, "Solid
Phase Peptide Synthesis" (Freeman & Co., San Francisco 1969),
Chapter 1, pp. 1-6. BHA and MBHA resin supports are commercially
available and are generally used only when the desired polypeptide
being synthesized has an a-carboxamide at the C-terminal.
[0048] Activating reagents used in solid phase synthesis of the
peptides are well known in the peptide synthesis art. Examples of
suitable activating reagents are (1) carbodiimides, such as
N,N-diisopropylcarbodiimide, N,N-dicyclohexylcarbodiimide (DCCI);
(2) cyanamides such as N,N-dibenzylcyanamide; (3) keteimines; (4)
isoxazolium salts, such as N-ethyl-5-phenyl
isoxazolium-3-sulfonate; (5) monocyclic nitrogen-containing
heterocyclic amides of aromatic character containing one through
four nitrogens in the ring, such as imidazolides, pyrazolides, and
1,2,4-triazolides. Specific heterocyclic amides that are useful
include N,N-carbonyl diimidazole, N,N-carbonyl-di-1,2,4-triazole;
(6) alkoxylated acetylene, such as ethoxyacetylene; (7) reagents
which form a mixed anhydride with the carboxyl moiety of the amino
acid, such as ethylchloroformate and isobutylchloroformate and (8)
reagents which form an active ester with the carboxyl moiety of the
amino acid, such as nitrogen-containing heterocyclic compounds
having a hydroxy group on one ring nitrogen, e.g.
N-hydroxyphthalimide, N-hydroxysuccinimide and
1-hydroxybenzotriazole (HOBT). Other activating reagents and their
use in peptide coupling are described by Schroder & Lubke
supra, in Chapter III and by Kapoor, J. PHAR. SCI., Vol. 59, pp.
1-27 (1970).
[0049] Each protected amino acid or amino acid sequence is
introduced into the solid phase reactor in about a two-fold or more
excess, and the coupling may be carried out in a medium of
dimethylformamide (DMF)--CH.sub.2 C.sub.12 (1:1) or in DMF or
CH.sub.2Cl.sub.2 alone. In cases where incomplete coupling occurs,
the coupling procedure is repeated before removal of the a-amino
protecting group prior to the coupling of the next amino acid. If
performed manually, the success of the coupling reaction at each
stage of the synthesis is monitored by the ninhydrin reaction, as
described by E. Kaiser et al., ANAL. BIOCHEM., Vol. 34, pp. 595
(1970).
[0050] After the desired amino acid sequence has been completed,
the intermediate peptide is removed from the resin support by
treatment with a reagent, such as liquid hydrogen fluoride, which
not only cleaves the peptide from the resin but also cleaves all
remaining side chain protecting groups X.sup.2, X.sup.3, X.sup.4,
X.sup.5, X.sup.1, X.sup.7, X.sup.8 and X.sup.9 and the
.alpha.-amino protecting group X.sup.1 to obtain the peptide.
[0051] As an alternative route, the intermediate peptide may be
separated from the resin support by alcoholysis after which the
recovered C-terminal alkyl ester is converted to the acid by
hydrolysis. Any side chain protecting groups may then be cleaved as
previously described or by other known procedures, such as
catalytic reduction (e.g. Pd on BaSO.sub.4). When using hydrogen
fluoride for cleaving, anisole and methylethyl sulphide are
included in the reaction vessel for scavenging.
[0052] For example, single peptides can be prepared advantageously
with the aid of an automatic MilliGen 9050 peptide synthesizer,
overlapping peptides with the aid of a multiple peptide synthesizer
SMPS 350 (ZINSSER Analytik) by means of solid-phase synthesis (R.
B. Merrifield (1966) J. Am. Chem. Soc. 85, 2149) following the
Fmoc/But strategy (E. Atherton, R. C. Sheppard; "Solid-Phase
Peptide Synthesis: A Practical Approach", IRL Press, Oxford,
England (1989).
[0053] Coupling and cleavage steps may be checked by means of
on-line UV monitoring. A substance which can advantageously be used
as solid-phase matrix for free peptides is, for example, a
p-alkoxybenzyl alcohol-modified polystyrene resin (S.-S. Wang; J.
Am. Chem. Soc. 95 (1973), p. 1328), as solid-phase matrix for
peptide amides Rink amide resin (H. Rink; Tetrahedron Lett. 28
(1987), p. 3787). TBTU
(2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium
tetrafluoroborate) may advantageously be employed as coupling
reagent (R. Knorr, A. Trzeciak, W. Bannwarth, D. Gillesen;
Tetrahedron Lett. 30 (1989), p. 1927). Usual coupling times are in
each 10-30, in particular approx. 20, minutes. After the synthesis
has ended, the peptides may be cleaved from the resin, for example
with the aid of trifluoroacetic acid and, if desired, added
substances (for example phenol, thioanisole, but also water) and
simultaneously freed from all protective groups.
[0054] The reaction solution may give a crude product, for example
by dropwise addition to ice-cold diethyl ether, which can be
purified, if desired, by washing with a suitable washing fluid, for
example cold diethyl ether, and then lyophilized. The crude peptide
is then subjected to ultrapurification to a purity of >98%,
advantageously by means of preparative reversed-phase
high-performance liquid chromatography (RP-HPLC) on a C18-column
(250.times.10 mm) using an acetonitrile/0.05% trifluoroacetic
acid/water system.
[0055] Peptides which are acylated by the N-terminus can be
obtained in high yields by reacting the N-terminally deprotected
peptides which are bound to the resin with acid chlorides or acid
anhydrides (for example palmitoyl chloride or lauryl chloride) in a
suitable solvent (for example
dimethylformamide/N-methylmorpholine).
[0056] The cosmetically or pharmaceutically acceptable
oligopeptides used in accordance with the invention, hereinbelow
also collectively termed "active ingredient used in accordance with
the invention", irrespective of whether an individual substance or
an isomer mixture or a mixture of a variety of individual
substances is present, have shown to be outstanding active
ingredients against undesired pigmentation, in particular local
hyperpigmentation, both preventatively and therapeutically.
[0057] The content of active ingredient used in accordance with the
invention in the cosmetic or topical dermatological preparations
may, in accordance with the invention, amount to 0.000001-10% by
weight, preferably 0.00001-1% by weight, in particular 0.0001-0.1%
by weight, based on the total weight of the preparations.
[0058] Surprisingly, it has emerged that the objects on which the
invention is based are achieved by the active ingredient used in
accordance with the invention. When using the active ingredient
used in accordance with the invention, or cosmetic or topical
dermatological preparations with an effective content of active
ingredient used in accordance with the invention, effective
prophylaxis against undesired pigmentation is possible. According
to the invention, it is, in particular, extremely advantageous to
use the active ingredient used in accordance with the invention, or
cosmetic or topical dermatological preparations with an effective
content of active ingredient used in accordance with the invention,
for the cosmetic or dermatological treatment of undesired skin
pigmentation, that is, for example, Lentigines seniles.
[0059] The prophylaxis and/or the cosmetic or dermatological
treatment with the active ingredient used in accordance with the
invention, or with the cosmetic or topical dermatological
preparations with an effective content of active ingredient used in
accordance with the invention, is effected in the customary manner
by applying the active ingredient used in accordance with the
invention, or the cosmetic or topical dermatological preparations
with an effective content of active ingredient used in accordance
with the invention, to the affected parts of the skin.
[0060] The active ingredient used in accordance with the invention
may advantageously be incorporated into customary cosmetic and
dermatological preparations, which may exist in a variety of forms.
For example, they may be a solution, a water-in-oil (W/O) type
emulsion, an oil-in-water (O/W) type emulsion, or a multiple
emulsion, for example a water-in-oil-in-water (W/O/W) or
oil-in-water-in-oil (O/W/O) emulsion, a hydrodispersion or
lipodispersion, a gel, a solid stick or else an aerosol.
[0061] Emulsions in accordance with the present invention, for
example in the form of a cream, a lotion or a cosmetic milk, are
advantageous and comprise, for example, fats, oils, waxes and/or
other lipids, as well as water and one or more emulsifiers as they
are usually used for such a type of formulation.
[0062] It is also possible and advantageous in accordance with the
present invention to incorporate the active ingredient used in
accordance with the invention into aqueous systems or surfactant
preparations for cleansing skin and hair.
[0063] Naturally, the expert knows that high-quality cosmetic
compositions are unthinkable without the customary adjuvants and
additives in most cases. These include, for example, thickeners,
fillers, fragrance, colours, emulsifiers, additional active
ingredients such as vitamins or proteins, sunscreens, stabilizers,
insect repellents, alcohol, water, salts, antimicrobially,
proteolytically or keratolytically active substances, and the
like.
[0064] The same also applies analogously to corresponding
requirements regarding the formulation of medicinal
preparations.
[0065] Medicinal topical compositions in accordance with the
present invention comprise, as a rule, one or more medicaments in
active concentration. For the sake of simplicity, reference is made
to the legal provisions of the Federal Republic of Germany to
differentiate neatly between cosmetic and medicinal use and
corresponding products (for example Kosmetikverordnung [cosmetics
regulation], Lebensmittelgesetz [food act] and Arzneimittelgesetz
[pharmaceuticals act]).
[0066] It is also advantageous to add the active ingredient used in
accordance with the invention to preparations which already
comprise other active ingredients for other purposes, as
additive.
[0067] Accordingly, cosmetic or topical dermatological compositions
in accordance with the present invention may be used, for example,
as a protective skin cream, cleansing milk, sun protection lotion,
nutrient cream, day cream or night cream and the like, depending on
their composition. If appropriate, it is possible and advantageous
to use the compositions according to the invention as basis for
pharmaceutical formulations.
[0068] If appropriate, other advantageous cosmetic and
dermatological preparations are those which are present in the form
of a sun protection product. These preferably comprise, in addition
to the active ingredient used in accordance with the invention, at
least one additional UVA filter and/or at least one UVB filter
and/or at least one inorganic pigment.
[0069] However, also advantageous in accordance with the present
invention is to create cosmetic and dermatological preparations
whose main purpose is not the protection from sunlight, but which
nevertheless comprise substances which protect from UV. For
example, UV-A and/or UV-B filters are usually incorporated into day
creams.
[0070] Preparations according to the invention may advantageously
comprise substances which absorb UV radiation in the UVB range, the
total amount of filters being, for example, 0.1% by weight to 30%
by weight, preferably 0.5 to 10% by weight, in particular 1 to 6%
by weight, based on the total weight of the preparations.
[0071] The UVB filters may be soluble in oil or in water. Examples
of substances which are soluble in oil are, for example: [0072]
3-benzylidenecamphor and its derivatives, for example
3-(4-methylbenzylidene)camphor, [0073] 4-aminobenzoic acid
derivatives, preferably 2-ethylhexyl 4-dimethylaminobenzoate, amyl
4-dimethylaminobenzoate; [0074] cinnamic esters, preferably
2-ethylhexyl 4-methoxycinnamate, isopentyl 4-methoxycinnamate;
[0075] salicylic esters, preferably 2-ethylhexyl salicylate,
4-isopropylbenzyl salicylate, homomethyl salicylate; [0076]
benzophenone derivatives, preferably
2-hydroxy-4-methoxybenzophenone,
2-hydroxy-4-methoxy-4'-methyl-benzophenone,
2,2'-dihydroxy-4-methoxybenzophenone; [0077] benzalmalonic esters,
preferably di(2-ethylhexyl) 4-methoxybenzalmalonate; [0078]
2,4,6-trianilino-(p-carbo-2'-ethyl-1'-hexyloxy)-1,3,5-triazine
[0079] Advantageous substances which are soluble in water are:
[0080] 2-phenylbenzimidazole-5-sulphonic acid and its salts, for
example sodium, potassium or triethanolammonium salts, [0081]
sulphonic acid derivatives of benzophenones, preferably
2-hydroxy-4-methoxybenzophenone-5-sulphonic acid and its salts;
[0082] sulphonic acid derivatives of 3-benzylidenecamphor such as,
for example, 4-(2-oxo-3-bornylidene-methyl)benzenesulphonic acid,
2-methyl-5-(2-oxo-3-bornylidenemethyl)sulphonic acid and their
salts.
[0083] Naturally, the list of the abovementioned UVB filters which
may be used according to the invention is not intended to be
limiting.
[0084] The invention also relates to the combination of a UVA
filter according to the invention with a UVB filter, and/or a
cosmetic or dermatological preparation according to the invention
which also comprises a UVB filter.
[0085] It may also be advantageous to employ, in preparations
according to the invention, UVA filters which cosmetic and/or
dermatological preparations usually comprise. Such filters are
preferably dibenzoylmethane derivatives, in particular
1-(4'-tert-butylphenyl)-3-(4'-methoxyphenyl)propane-1,3-dione and
1-phenyl-3-(41-isopropylphenyl)propane-1,3-dione. The invention
also relates to preparations which comprise these combinations. The
same amounts of UVA filters which have been mentioned for UVB
filters may be used.
[0086] Cosmetic and/or dermatological preparations in accordance
with the present invention may also comprise inorganic pigments
which are usually used in cosmetology for protecting the skin from
UV rays. They are oxides of titanium, zinc, iron, zirconium,
silicon, manganese, aluminium, cerium and mixtures of these, and
modifications where the oxides are the active agents. Especially
preferably, they are pigments based on titanium dioxide. The
amounts mentioned for the above combinations may be used.
[0087] The cosmetic and dermatological preparations according to
the invention may comprise cosmetically active ingredients,
cosmetic auxiliaries and/or cosmetic additives conventionally used
in such preparations, for example antioxidants, preservatives,
bactericides, fragrances, antifoams, colorants, colour pigments,
thickeners, surfactants, emulsifiers, plasticizers, moisturizers
and/or humectants, fats, oils, waxes or other customary
constituents of a cosmetic or dermatological formulation such as
alcohols, polyols, polymers, foam stabilizers, electrolytes,
organic solvents or silicone derivatives.
[0088] It is also advantageous to add customary antioxidants to the
preparations in accordance with the present invention. Advantageous
antioxidants which may be used in accordance with the invention are
all those antioxidants which are suitable or conventional for
cosmetic and/or dermatological applications.
[0089] The antioxidants are advantageously selected from the group
consisting of amino acids (for example glycine, histidine,
tyrosine, tryptophan) and their derivatives, imidazoles (for
example urocaninic acid) and their derivatives, peptides such as
D,L-carnosine, D-carnosine, L-carnosine and their derivatives (for
example anserine), carotenoids, carotenes (for example
.alpha.-carotene, .beta.-carotene, lycopine) and their derivatives,
liponic acid and its derivatives (for example dihydrolipoic acid),
aurothioglucose, propylthiouracil and other thiols (for example
thioredoxin, glutathione, cysteine, cystine, cystamine and their
glycosyl, N-acetyl, methyl, ethyl, propyl, amyl, butyl and lauryl,
palmitoyl, oleyl, .gamma.-linoleyl, cholesteryl and glyceryl
esters) and their salts, dilauryl thiodipropionate, distearyl
thiodipropionate, thiodipropionic acid and its derivatives (esters,
ethers, peptides, lipids, nucleotides, nucleosides and salts) and
sulphoximine compounds (for example buthionine sulphoximines,
homocysteine sulphoximine, buthionine sulphones, penta-, hexa-,
heptathionine sulphoximine) at very low tolerated doses (for
example pmol to .mu.mol/kg), furthermore (metal)chelating agents
(for example .alpha.-hydroxy fatty acids, palmitic acid, phytic
acid, lactoferrin), .alpha.-hydroxy acids (for example citric acid,
lactic acid, malic acid), humic acid, bile acid, bile extracts,
bilirubin, biliverdin, EDTA, EGTA and their derivatives,
unsaturated fatty acids and their derivatives (for example
.gamma.-linolenic acid, linolic acid, oleic acid), folic acid and
its derivatives, alaninediacetic acid, flavonoids, polyphenols,
catechols, ubiquinone and ubiquinol and their derivatives, vitamin
C and derivatives (for example ascorbyl palmitate, Mg-ascorbyl
phosphate, ascorbyl acetate), tocopherols and derivatives (for
example vitamin E acetate), and coniferyl benzoate of benzoin
resin, rutinic acid and its derivatives, ferulic acid and its
derivatives, butylhydroxytoluene, butylhydroxyanisole,
nordihydroguaiacic acid, nordihydroguairetic acid,
trihydroxybutylrophenone, uric acid and its derivatives, mannose
and its derivatives, zinc and its derivatives (for example ZnO,
ZnSO.sub.4) selenium and its derivatives (for example selenium
methionine), stilbene and its derivatives (for example stilbene
oxide, trans-stilbene oxide) and those derivatives of the
abovementioned active ingredients which are suitable according to
the invention (salts, esters, ethers, sugars, nucleotides,
nucleosides, peptides and lipids).
[0090] The amount of antioxidants (one or more compounds) in the
preparations is preferably 0.001 to 30% by weight, especially
preferably 0.05-20% by weight, in particular 1-10% by weight, based
on the total weight of the preparation.
[0091] If the antioxidant(s) is/are vitamin E and/or its
derivatives, it is advantageous to choose the respective
concentrations from the range 0.001-10% by weight, based on the
total weight of the formulation.
[0092] If the cosmetic or dermatological preparation according to
the present invention represents a solution or emulsion or
dispersion, the following may be used as solvents: [0093] water or
aqueous solutions; [0094] oils such as triglycerides of capric or
caprylic acid, preferably castor oil; [0095] fats, waxes and other
natural and synthetic lipids, preferably esters of fatty acids with
alcohols of low C number, for example with isopropanol, propylene
glycol or glycerol, or esters of fatty alcohols with alkanolic
acids of low C number or with fatty acids; [0096] alcohols, diols
or polyols of low C number and their ethers, preferably ethanol,
isopropanol, propylene glycol, glycerol, ethylene glycol, ethylene
glycol monoethyl ether or ethylene glycol monobutyl ether,
propylene glycol monomethyl ether, propylene glycol monoethyl ether
or propylene glycol monobutyl ether, diethylene glycol monomethyl
ether or diethylene glycol monoethyl ether, and analogous
products.
[0097] In particular, mixtures of the abovementioned solvents are
used. In the case of alcoholic solvents, water may be a further
constituent.
[0098] The oil phase of the emulsions, oleogels or hydro- or
lipodispersions in accordance with the present invention is
advantageously selected from the group of the esters of saturated
and/or unsaturated, branched and/or unbranched alkanecarboxylic
acids with a chain length of 3 to 30 C atoms and saturated and/or
unsaturated branched and/or unbranched alcohols with a chain length
of 3 to 30 C atoms, from the group of the esters of aromatic
carboxylic acids and saturated and/or unsaturated, branched and/or
unbranched alcohols with a chain length of 3 to 30 C atoms. In this
case, such ester oils may be selected advantageously from the group
consisting of isopropyl myristate, isopropyl palmitate, isopropyl
stearate, isopropyl oleate, n-butyl stearate, n-hexyl laurate,
n-decyl oleate, isooctyl stearate, isononyl stearate, isononyl
isononanoate, 2-ethylhexyl palmitate, 2-ethylhexyl laurate,
2-hexyl-decyl stearate, 2-octyldodecyl palmitate, oleyl oleate,
oleyl erucate, erucyl oleate, erucyl erucate, and synthetic,
semisynthetic and natural mixtures of such esters, for example
jojoba oil.
[0099] Furthermore, the oil phase may advantageously be selected
from the group of the branched and unbranched hydrocarbons and
hydrocarbon waxes, the silicone oils, the dialkyl ethers, the group
of the saturated or unsaturated branched or unbranched alcohols and
of the fatty acid triglycerides, viz. the triglycerol esters of
saturated and/or unsaturated, branched and/or unbranched
alkanecarboxylic acids with a chain length of 8 to 24, in
particular 12-18, C atoms. For example, the fatty acid
triglycerides may advantageously be selected from the group of the
synthetic, semisynthetic and natural oils, for example olive oil,
sunflower oil, soya oil, peanut oil, rapeseed oil, almond oil, palm
oil, coconut oil, palm kernel oil and the like.
[0100] Any mixtures of such oil and wax components may also
advantageously be employed in accordance with the present
invention. If appropriate, it may also be advantageous to employ
waxes, for example cetyl palmitate, as the only lipid component of
the oil phase.
[0101] The oil phase is advantageously selected from the group
consisting of 2-ethylhexyl isostearate, octyldodecanol, isotridecyl
isononanoate, isoeicosan, 2-ethylhexyl cocoate, C.sub.12-15-alkyl
benzoate, caprylic/capric acid triglyceride, dicaprylyl ether.
[0102] Especially advantageous mixtures are those of
C.sub.12-15-alkyl benzoate and 2-ethylhexyl isostearate, those of
C.sub.12-15-alkyl benzoate and isotridecyl isononanoate and those
of C.sub.12-15-alkyl benzoate, 2-ethylhexyl isostearate and
isotridecyl isononanoate.
[0103] Amongst the hydrocarbons, liquid paraffin, squalane and
squalene may advantageously be used according to the present
invention.
[0104] The oil phase may furthermore advantageously comprise cyclic
or linear silicone oils, or consist entirely of such oils, but it
is preferred to use an additional content of another oil phase
components, apart from the silicone oil(s).
[0105] Cyclomethicone (octamethylcyclotetrasiloxane) is
advantageously employed as silicone oil to be used according to the
invention. However, other silicone oils may also be used
advantageously in accordance with the present invention, for
example hexamethylcyclotrisiloxane, polydimethylsiloxane,
poly(methylphenylsiloxane).
[0106] Especially advantageous mixtures are furthermore those of
cyclomethicone and isotridecyl isononanoate and of cyclomethicone
and 2-ethylhexyl isostearate.
[0107] If appropriate, the aqueous phase of the preparations
according to the invention advantageously comprises [0108]
alcohols, diols or polyols of low C number, and their ethers,
preferably ethanol, isopropanol, propylene glycol, glycerol,
ethylene glycol, ethylene glycol monoethyl ether or ethylene glycol
monobutyl ether, propylene glycol monomethyl ether, propylene
glycol monoethyl ether or propylene glycol monobutyl ether,
diethylene glycol monomethyl ether or diethylene glycol monoethyl
ether and analogous products, furthermore alcohols of low C number,
for example ethanol, isopropanol, 1,2-propanediol, glycerol, and,
in particular, one or more thickeners which may advantageously be
selected from the group consisting of silicon dioxide, aluminium
silicates, polysaccharides and their derivatives, for example
hyaluronic acid, xanthan gum, hydroxy-propylmethylcellulose,
especially advantageously from the group of the polyacrylates,
preferably a polyacrylate from the group of the so-called
Carbopols, for example type 980, 981, 1382, 2984 and 5984
Carbopols, in each case singly or in combination.
[0109] Gels used according to the invention usually comprise
alcohols of low C number, for example ethanol, isopropanol,
1,2-propanediol, glycerol and water, or an abovementioned oil in
the presence of a thickener, which is preferably silicon dioxide or
an aluminium silicate in the case of oily-alcoholic gels and
preferably a polyacrylate in the case of aqueous-alcoholic or
alcoholic gels.
[0110] Solid sticks comprise, for example, natural or synthetic
waxes, fatty alcohols or fatty acid esters.
[0111] Customary basic materials which are suitable for use as
cosmetic sticks in accordance with the present invention are liquid
oils (for example liquid paraffin, castor oil, isopropyl
myristate), semi-solid constituents (for example petrolatum,
lanolin), solid constituents (for example beeswax, ceresine and
micro-crystalline waxes, or ozocerite) and waxes of high melting
point (for example carnauba wax, candelilla wax).
[0112] Suitable propellants for cosmetic and/or dermatological
preparations in accordance with the present invention which can be
sprayed from aerosol containers are the customary known volatile,
liquefied propellants, for example hydrocarbons (propane, butane,
isobutane), which may be employed singly or as a mixture with each
other. Pressurized air may also be used advantageously.
[0113] The person skilled in the art will, of course, be familiar
with the fact that there are non-toxic propellants, which would be
suitable in principle for putting into practice the present
invention in the form of aerosol preparations; however, it is
recommended to manage without these--in particular
fluorohydrocarbons and fluorochlorohydrocarbons (FCHCs)--due to
their unacceptable effect on the environment or other accompanying
circumstances.
[0114] Cosmetic preparations in accordance with the present
invention may also be present as gels which comprise not only an
effective amount of active ingredient according to the invention
and conventionally used solvents therefor, preferably water, but
also organic thickeners, for example gum arabic, xanthan gum,
sodium alginate, cellulose derivatives, preferably
methyl-cellulose, hydroxymethylcellulose, hydroxyethylcellulose,
hydroxypropylcellulose, hydroxypropylmethyl-cellulose, or inorganic
thickeners, for example aluminium silicates such as, for example,
bentonites, or a mixture of polyethylene glycol and polyethylene
glycol stearate or polyethylene glycol distearate. The gel
comprises the thickener for example in an amount of between 0.1 and
30% by weight, preferably between 0.5 and 15% by weight.
[0115] The examples which follow are intended to illustrate the
present invention.
PREPARATION EXAMPLE 1
Synthesis in a Reaction Vessel (Suitable for Larger Batches)
[0116] In a reaction vessel, 0.5 g of Rink amide resin (0.2 mmol/g)
are washed thoroughly with DMF and then treated with 5 ml of 20%
(v/v) piperidine/DMF solution. After 3 minutes, the batch is
filtered and is treated for a further 5 minutes with 5 ml of 20%
piperidine/DMF. After thorough washing with DMF (approx. 50 ml),
the resin is treated with 108.3 mg of Fmoc-Pro-OH and 128.1 mg of
TBTU in 2 ml of DMF. After addition of 1.25 ml of 0.7 M
N-methylmorpholine solution, the mixture is shaken for 30 minutes.
After washing of the resin, completeness of the reaction is checked
by a Kaiser test.
[0117] After the Fmoc protective group has been eliminated using
20% piperidine/DMF, 259.5 mg of Fmoc-Arg(Pbf)-OH (0-4 mmol), 135.8
mg (0.4 mmol) of Fmoc-Val-OH and a further 135.8 mg (0.4 mmol) of
Fmoc-Val-QH are coupled as described in the scheme above. Each
time, the coupling efficiency is checked with the aid of the Kaiser
test. After deblocking of the Fmoc protective group, the resin is
washed thoroughly with DMF and finally dried. The protective group
and the peptide are cleaved from the resin by 2 ml of
trifluoroacetic acid/thioanisole/phenol/water 90:5:3:2 (v,v). After
the mixture has reacted for 2 hours, it is filtered and the
filtrate is directly added dropwise to 30 ml of ice-cooled diethyl
ether. After centrifugation, the precipitate is washed thoroughly
with diethyl ether and recentrifuged. The procedure is repeated
three times. After dissolving the pellet in tert.-butanol/water
(4:1, v/v), the solution is lyophilized.
[0118] The colourless lyophilisate is taken up in 5 ml of deionized
water and again freeze-dried.
PREPARATION EXAMPLE 2
Synthesis of H--VVRP--NH.sub.2 (Synthesis in Synthesizer)
[0119] A high-pressure glass column of the MilliGen 9050 peptide
synthesizer is packed with 0.5 g of Rink amide resin (0.2 mmol/g)
to which 2.5 g of glass beads (0.150-212 .mu.m, Sigma) had been
admixed. The N-terminal Fmoc protective group is then removed by
adding 14.8 ml of 20% (v/v) piperidine/dimethylformamide (flow rate
7.4 ml/min). After washing with dimethylformamide (5 min at 7.4
ml/min), 108.3 mg (0.4 mmol) of Fmoc-proline and 128 mg of TBTU
(0.4 mmol) in 1.25 ml of 0.7 M N-methylmorpholine solution are
dissolved in DMF. After 3 minutes, this preactivated solution is
pumped onto the column by the recycling method and coupled for 20
minutes. After washing with DMF, the N.sup..alpha.-Fmoc group is
again eliminated by adding 20% piperidine in DMF. 259.5 mg of
Fmoc-Arg(Pbf)-OH (0.4 mmol) are coupled as above as the next amino
acid derivative. After coupling Fmoc-Val-OH and a further
Fmoc-Val-OH (in each case 135.8 mg, 0.4 mmol), the Fmoc group is
again eliminated, and the resin is washed thoroughly with DMF,
removed from the column and dried thoroughly. The protective group
and the peptide are cleaved from the resin by 2 ml of
trifluoroacetic acid/thioanisole/phenol/water 90:5:3:2 (v/v). After
the mixture has reacted for 2 hours, it is filtered and the
filtrate is directly added dropwise to 30 ml of ice-cooled diethyl
ether. After centrifugation, the precipitate is washed thoroughly
with diethyl ether and recentrifuged. The procedure is repeated
three times. After dissolving the pellet in tert.-butanol/water
(4:1, v/v), the solution is lyophilized. The colourless
lyophilisate is again taken up in 5 ml of deionized water and again
freeze-dried.
ABBREVIATIONS
[0120] Fmoc: 9-fluorenylmethoxycarbonyl residue [0121] Rink amide
resin: 4-(2',4'-dimethylphenyl-Fmoc-aminomethyl)-phenoxy resin
[0122] Fmoc-Arg(Pbf)-OH:
N.sup..alpha.-Fmoc-N.sup.G-2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulph-
onyl-arginine [0123] TBTU:
2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium
tetrafluoroborate [0124] DMF dimethylformamide
EXAMPLE 1
TABLE-US-00002 [0125] W/O cream % by weight Liquid paraffin (DAB 9)
10.00 Petrolatum 4.00 Wool wax alcohol 1.00 PEG-7-hydrogenated
castor oil 3.00 Aluminium stearate 0.40 H-VVRP-NH.sub.2 0.01
Glycerol 2.00 Preservatives, colours, fragrance q.s. Water to
100.00
EXAMPLE 2
TABLE-US-00003 [0126] W/O lotion % by weight Liquid paraffin (DAB
9) 20.00 Petrolatum 4.00 Glucose sesquiisostearate 2.00 Aluminium
stearate 0.40 H-VVRP-NH.sub.2 0.005 .alpha.-Tocopheryl acetate 1.00
Glycerol 5.00 Preservatives, colours, fragrance q.s. Water to
100.00
EXAMPLE 3
TABLE-US-00004 [0127] O/W lotion % by weight Liquid paraffin (DAB
9) 8.00 Isopropyl palmitate 3.00 Petrolatum 4.00 Cetylstearyl
alcohol 2.00 PEG 40 castor oil 0.50 Sodium cetyl stearyl sulphate
0.50 Sodium carbomer 0.40 H-VVRP-NH.sub.2 0.001 Glycerol 3.00
.alpha.-Tocopherol 0.20 Octyl methoxycinnamate 5.00
Butylmethoxydibenzoylmethane 1.00 Preservatives, colours, fragrance
q.s. Water to 100.00
EXAMPLE 4
TABLE-US-00005 [0128] O/W cream % by weight Liquid paraffin (DAB 9)
7.00 Avocado oil 4.00 Glyceryl monostearate 2.00
Acetyl-VVRP-NH.sub.2 0.01 Titanium dioxide 1.00 Sodium lactate 3.00
Glycerol 3.00 Preservatives, colours, fragrance q.s. Water to
100.00
EXAMPLE 5
TABLE-US-00006 [0129] Liposome-containing gel % by weight Lecithin
6.00 Shea butter 3.00 H-VVRP-NH.sub.2 0.005 .alpha.-Tocopherol 0.20
Biotin 0.08 Sodium citrate 0.50 Glycine 0.20 Urea 0.20 Sodium PCA
0.50 Hydrolysed collagen 2.00 Xanthan gum 1.40 Sorbitol 3.00
Preservatives, colours, fragrance q.s. Water to 100.00
EXAMPLE 6
TABLE-US-00007 [0130] Gel % by weight Carbopol 934 P 2.00
Triethanolamin 3.00 Acetyl-VVRP-NH.sub.2 0.001 .alpha.-Tocopheryl
acetate 0.20 Polyoxyethylene sorbitan fatty 0.50 acid ester (Tween
20) Glycerol 2.00 Sodium PCA 0.50 Hydrolysed collagen 2.00
Preservatives, colours, fragrance q.s. Water to 100.00
EXAMPLE 7
TABLE-US-00008 [0131] Sun protection emulsion % by weight
Cyclomethicone 2.00 Cetyldimethicone copolyol 0.20 PEG 22-Dodecyl
copolymer 3.00 Liquid paraffin (DAB 9) 2.00 Caprylic/capric acid
triglyceride 5.80 Octyl methoxycinnamate 5.80
Butylmethoxydibenzoylmethane 4.00 Acetyl-VVRP 0.001
.alpha.-Tocopheryl acetate 0.50 ZnSO.sub.4 0.70 Na.sub.4EDTA 0.30
Preservatives, colours, fragrance q.s. Water to 100.00
EXAMPLE 8
TABLE-US-00009 [0132] Sun protection emulsion % by weight
Cyclomethicone 2.00 Cetylstearyl alcohol + PEG 40 2.50 hydrogenated
castor oil + sodium cetyl stearyl sulphate Glyceryl lanolate 1.00
Caprylic/capric acid triglyceride 0.10 Laurylmethicone copolyol
2.00 Octyl stearate 3.00 Castor oil 4.00 Glycerol 3.00
Acrylamide/sodium acrylate 0.30 copolymer
Hydroxypropylmethylcellulose 0.30 Octyl methoxycinnamate 5.00
Butylmethoxydibenzoylmethane 0.50 H-VVRP-NH.sub.2 0.001
.alpha.-Tocopheryl acetate 1.00 Na.sub.3HEDTA 1.50 Preservatives,
colours, fragrance q.s. Water to 100.00
EXAMPLE 9
TABLE-US-00010 [0133] Sun protection emulsion % by weight
Cyclomethicone 2.00 Cetylstearyl alcohol + PEG 40 2.50 hydrogenated
castor oil + sodium cetyl stearyl sulphate Glyceryl lanolate 1.00
Caprylic/capric acid triglyceride 0.10 Laurylmethicone copolyol
2.00 Octyl stearate 3.00 Castor oil 4.00 Glycerol 3.00
Acrylamide/sodium acrylate 0.30 copolymer
Hydroxypropylmethylcellulose 0.30 Octyl methoxycinnamate 5.00
Butylmethoxydibenzoylmethane 0.75 Acetyl-VVRP-NH.sub.2 0.001
Na.sub.3HEDTA 1.50 Preservatives, colours, fragrance q.s. Water to
100.00
EXAMPLE 10
TABLE-US-00011 [0134] Massage cream % by weight Stearyl alcohol
2.00 Petrolatum 4.00 Dimethicone 2.00 Isopropyl palmitate 6.00
Cetyl stearyl alcohol 4.00 PEG 40 hydrogenated castor oil 2.00
.alpha.-Tocopherol 0.50 H-VVRP-NH.sub.2 0.005 Glycerol 3.00
Preservatives, colours, fragrance q.s. Water to 100.00
EXAMPLE 11
TABLE-US-00012 [0135] Hair tonic % by weight Ethanol 40.00
Diisopropyl adipate 0.10 PEG 40 hydrogenated castor oil 0.20
H-VVRP-NH.sub.2 0.001 .alpha.-Tocopheryl acetate 0.10
Preservatives, colours, fragrance q.s. Water to 100.00
EXAMPLE 12
TABLE-US-00013 [0136] Spray formulation % by weight
.alpha.-Tocopherol 0.10 Acetyl-VVRP-NH.sub.2 0.001 Ethanol 28.20
Preservatives, colours, fragrance q.s. Propane/butane 25/75 to
100.00
Sequence CWU 1
1
314PRTArtificial SequenceDescription of Artificial Sequence
structural base for oligopeptides 1Val Val Arg Pro 125PRTArtificial
SequenceDescription of Artificial Sequence structural base for
oligopeptides 2Val Val Arg Pro Pro 1 536PRTArtificial
SequenceDescription of Artificial Sequence Structural base for
oligopeptides. 3Val Val Arg Pro Pro Pro 1 5
* * * * *