Substrate-specific adhesive peptides

Abstract

The invention relates to peptides comprising or consisting of an amino acid sequence of 5 to 50 amino acids which has, in N- to C-terminus orientation, the amino acid sequence X5X6X7X8X9, where each of the five residues X represents an amino acid of a particular group, and / or the amino acid sequence has at least 85% sequence identity to one of the amino acid sequences mentioned in SEQ ID NOs: 1 - 17.

Description

[0001] Substrate-specific adhesive peptides

[0002] The present application relates to peptides with a specific binding capacity to keratin fibers. The peptides according to the invention comprise or consist of an amino acid sequence of 5 to 50 amino acids, characterized in that the amino acid residues at positions X5, X6, X7, X8, and X9 are each selected from a group of very specific amino acids, and / or that the peptides have a sequence identity of at least 85% with the amino acid sequences listed in SEQ ID NOs: 1-17.

[0003] Further subjects of the present application are the nucleic acid sequences which encode the corresponding peptides, an agent for treating keratinic fibers, in particular human hair, which agent contains at least one of the peptides according to the invention, and a method for treating keratinic fibers, in which an agent which contains at least one peptide according to the invention is applied to the keratinic fibers and optionally rinsed out again after a contact time.

[0004] In the field of hair cosmetics, the surface treatment of keratin fibers is a central topic. The defined deposition of substances on the hair surface can, for example, increase its diameter and thus its volume; conditioning ingredients can form a barrier against external influences; or the formation of films can improve combability and manageability. In recent decades, a wide range of different substance classes have been used to specifically modify the hair surface, ranging from polymers and emulsifiers to cationic surfactants and monomeric active ingredients. As the topic of sustainability has recently become increasingly important, the current focus is primarily on new compounds of biological origin.Adhesive peptides are of particular interest in this context because they are not only biodegradable, but as chains of linked amino acids they also closely resemble the composition of keratin fibers.

[0005] For hair treatment, the main focus is on finding new peptides that bind specifically to the surface of keratin fibers and remain there for extended periods, but have little or no substantivity on other substrates such as textiles or skin. By binding to the hair surface, such peptides can specifically modify the structure of the surface or be used as anchor molecules to bind other active ingredients to the surface. The objective of this application was therefore to find new peptides with the highest possible substantivity or specific binding capacity to keratin fibers, particularly human hair. The new peptides should adhere as well and as long as possible to different hair types. Furthermore, the peptides should develop less substantivity or adhesive capacity on other substrates such as textiles or skin.

[0006] During the development work leading to this invention, it was discovered that the special peptides described below surprisingly have particularly good adhesion to keratin fibers.

[0007] A first subject of the present application is therefore a peptide comprising or consisting of an amino acid sequence of 5 to 50 amino acids, wherein

[0008] (a) the amino acid sequence in N- to C-terminal orientation has the following sequence, (X a )nX5X6X7X8X9(Xb)m where each X a independently of one another is any amino acid, n is an integer from 0 to 45, preferably from 0 to 4,

[0009] X5 is an amino acid selected from the group consisting of P, H, T, N, R, C and A,

[0010] Xe is an amino acid selected from the group consisting of T, R, M, G, F, P, N, S, L, Y and W,

[0011] X7 is an amino acid selected from the group consisting of R, N, S, L, I, Q, W, G, A and Y,

[0012] Xe is an amino acid selected from the group consisting of K, T, N, A, L, F, D, R and G,

[0013] X9 is an amino acid selected from the group consisting of R, L, I, A, Q, P, H, G, W and M, each Xb is independently any amino acid, and m is an integer from 0 to 45, preferably from 0 to 7, particularly preferably from 0 to 3, with the proviso that when X5 stands for P, X9 does not simultaneously stand for P, and / or b) the amino acid sequence has at least 85%, preferably at least 86%, more preferably at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with one of the amino acid sequences mentioned in SEQ ID NOs: 1 - 17.

[0014] Peptides A peptide in the context of the present invention is understood to be a preferably linear polymer composed of amino acids, preferably the 20 proteinogenic L-amino acids, which comprises 5 to 50 amino acids linked to one another via peptide bonds. According to the invention, the peptides of the invention have an amino acid sequence of 5 to 50 amino acids. The amino acids are given in the context of this invention in a one-letter code, where, for example, C stands for cysteine, R for arginine, A for alanine and L for leucine. It is further understood that, unless otherwise stated, the amino acids in an amino acid sequence disclosed herein are linked via peptide bonds and, unless otherwise stated, the sequence is listed in N- to C-terminal orientation.

[0015] The peptides of the invention can, in various embodiments, be chemically synthesized and / or produced recombinantly by protein design. Short peptides are now easily synthesized, for example, by solid-phase synthesis. Longer peptides and polypeptides, on the other hand, are often also produced recombinantly in the host organism.

[0016] In the context of the present invention, the term “N-terminus” or “N-terminal” describes the end of the amino acid chain of the peptide according to the invention, which has a free amino group.

[0017] In the context of the present invention, the term “C-terminus” or “C-terminal” describes the end of the amino acid chain of the peptide according to the invention which has a free carboxyl group.

[0018] The term “in N- to C-terminal orientation” in the context of this invention refers to an amino acid sequence in which the order of the amino acids is described from the N-terminus to the C-terminus.

[0019] Typical acidic or negatively charged amino acids (depending on the pH value) are D and E.

[0020] Positively charged or basic amino acids (depending on pH) typically include R, K, and H.

[0021] Amino acids such as G, A, C, I, L, M, F, V, P, S, T, W, Y, N and Q are typically uncharged, i.e. neutral, amino acids.

[0022] When reference is made herein to an "any" amino acid, this typically means one of the 20 naturally occurring proteinogenic amino acids, i.e., one of glycine (G), alanine (A), valine (V), leucine (L), isoleucine (I), phenylalanine (F), serine (S), threonine (T), proline (P), methionine (M), cysteine ​​(C), histidine (H), lysine (K), arginine (R), glutamine (Q), asparagine (N), aspartic acid (D), glutamic acid (E), tyrosine (Y), and tryptophan (W). The amino acids are typically L-amino acids unless otherwise stated. In alternative embodiments, the peptide may also consist of D-amino acids, although it may be preferred that D- and L-amino acids do not occur simultaneously within the peptides described herein.

[0023] The peptide(s) according to the invention comprise or consist of an amino acid sequence of 5 to 50 amino acids.

[0024] Peptides consisting of an amino acid sequence of 5 to 50 amino acids have a chain that is 5 to 50 amino acids long. Peptides comprising an amino acid sequence of 5 to 50 amino acids have a chain of at least 5 amino acids to at least 50 amino acids, but can also have an even higher molecular weight and comprise a chain that is more than 50 amino acids long.

[0025] Preferably, the peptide has an amino acid sequence which has a length of 10 to 24 amino acids, for example 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 amino acids, in particular 8 to 18 amino acids, most preferably 12 amino acids.

[0026] In principle, better adhesion to the hair surface was observed with the shorter peptides compared to the longer peptides. It is suspected that the defined sequence X5X6X7X8X9, or the positions directly adjacent to this sequence described below, is one of the sites through which the peptide interacts with specific positions or amino acid sequences on the hair surface, so that the adhesion to the hair surface increases the larger the weight fraction of the interacting amino acids in relation to the total weight of the peptide.

[0027] Therefore, it is particularly preferred if the peptide according to the invention consists of an amino acid sequence of 5 to 50 amino acids, where

[0028] (a) the amino acid sequence in N- to C-terminal orientation has the following sequence, (X a )nX5X6X7X8X9(Xb)m where each X aindependently of one another is any amino acid, n is an integer from 0 to 45, preferably from 0 to 4,

[0029] X5 is an amino acid selected from the group consisting of P, H, T, N, R, C and A,

[0030] Xe is an amino acid selected from the group consisting of T, R, M, G, F, P, N, S, L, Y and W,

[0031] X7 is an amino acid selected from the group consisting of R, N, S, L, I, Q, W, G, A and Y, X8 is an amino acid selected from the group consisting of K, T, N, A, L, F, D, R and G,

[0032] X9 is an amino acid selected from the group consisting of R, L, I, A, Q, P, H, G, W and M, each Xb is independently any amino acid, and m is an integer from 0 to 45, preferably from 0 to 7, more preferably from 0 to 3, with the proviso that when X5 is P, X9 does not simultaneously also represent P.

[0033] The invention relates to a peptide comprising or consisting of an amino acid sequence of 5 to 50 amino acids in length, preferably of at least 8, 9, 10, 11, or 12 amino acids in length. Preferred lengths are 8 to 40, preferably 9 to 35, more preferably 10 to 30, and particularly preferably 11 to 25 amino acids. For example, the peptide can also have a length of 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 amino acids, in particular 12 to 18 amino acids.

[0034] Explicitly particularly preferably, the peptide consists of an amino acid sequence which has a length of 10 to 24 amino acids, for example 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 amino acids, in particular 8 to 18 amino acids, very particularly preferably 12 amino acids.

[0035] For example, if the peptide comprises at least 5 amino acids, then in embodiment (a) it is characterized in that it has an amino acid sequence in N- to C-terminal orientation,

[0036] (X a )nX5X6X7X8X9(Xb)m where n stands for the number 0, the residues X5, X6, X7, X8 and X9 each stand for an amino acid selected from the aforementioned specific groups, and m also stands for the number 0.

[0037] If the peptide consists of an amino acid sequence of 5 amino acids, then it consists of the above-mentioned chain of the 5 defined amino acids XsXeX XsXg.

[0038] If the peptide comprises at least 6 amino acids, then in embodiment (a) it is characterized in that it has an amino acid sequence in N- to C-terminal orientation, (X a)nX5X6X7X8X9(Xb)m where n is the number 0 or 1, the residues X5, X6, X7, X8 and X9 each represent an amino acid selected from the aforementioned specific groups, and m is the number 0 or 1, the sum of n+m being 1. The peptide then comprises either a residue Xa or a residue Xb. If the peptide consists of an amino acid sequence of 6 amino acids, then it consists of the above-mentioned chain of 6 defined amino acids, and therefore has or comprises the chain X a X5X6X7X8X9 or nX5X6X7X8X9Xb If the peptide comprises at least 50 amino acids, then in embodiment (a) it is characterized in that it has an amino acid sequence in the N- to C-terminal orientation, (X a)nX5X6X7X8X9(Xb)m where n is an integer from 0 to 45, residues X5, X6, X7, X8, and X9 each represent an amino acid selected from the aforementioned specific groups, and m is a number from 0 to 45, the sum of n+m being 45. The peptide then comprises a total of 45 residues Xa and Xb, where each Xa and each Xb independently represents any amino acid. This means that the 45 amino acid residues are located before and / or after the sequence X5X6X7X8X9.

[0039] Peptides, which include or consist of an amino acid sequence of 7 to 49 amino acids, are constructed in a similar way.

[0040] The residue Xa stands independently for any amino acid, ie Xa stands, for example, for an amino acid from the group of glycine (G), alanine (A), valine (V), leucine (L), isoleucine (I), phenylalanine (F), serine (S), threonine (T), proline (P), methionine (M), cysteine ​​(C), histidine (H), lysine (K), arginine (R), glutamine (Q), asparagine (N), aspartic acid (D), glutamic acid (E), tyrosine (Y) and tryptophan (W).

[0041] The index number n stands for an integer from 0 to 45. Preferably, n stands for an integer from 0 to 4. This means that in front of the chain of amino acids X5X6X7X8X9 (i.e. in the direction of the N-terminal end of the amino acid sequence) there is particularly preferably either no further amino acid or that 1 to 3 further amino acids are located there. If, for example, there are three further amino acids in front of the chain of amino acids X5X6X7X8X9, then the peptide in N- to C-terminal orientation comprises the sequence

[0042] XaX a X a X5X6X7X8X9(Xb)m

[0043] X5 is an amino acid selected from the group consisting of P (proline), H (histidine), T (threonine), N (asparagine), R (arginine), C (cysteine), and A (alanine). It has been found that peptides in particular possess particularly high substantivity to the surface of the keratin fiber, where X5 is an amino acid selected from the group consisting of P, N, and C, particularly preferably P.

[0044] In a particularly preferred embodiment, a peptide according to the invention is therefore characterized in that

[0045] (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (X a )nX5X6X7X8X9(Xb)m where Xs is an amino acid selected from the group consisting of P, N, and C, particularly preferably P.

[0046] Xe is an amino acid selected from the group consisting of T (threonine), R (arginine), M (methionine), G (glycine), F (phenylalanine), P (proline), N (asparagine), S (serine), L (leucine),

[0047] Y (tyrosine) and W (tryptophan).

[0048] In this context, it was found that especially those peptides have a particularly high substantivity to the surface of the keratin fiber in which Xe is an amino acid selected from the group consisting of T, R, M and P, preferably from the group consisting of T, R and M, particularly preferably from the group consisting of T and R.

[0049] In a particularly preferred embodiment, a peptide according to the invention is therefore characterized in that

[0050] (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (X a )nX5X6X7X8X9(Xb)m where

[0051] Xe is an amino acid selected from the group consisting of T, R, M and P, preferably from the group consisting of T, R and M, particularly preferably from the group consisting of T and R.

[0052] X7 is an amino acid selected from the group consisting of R (arginine), N (asparagine), S (serine), L (leucine), I (isoleucine), Q (glutamine), W (tryptophan), G (glycine), A (alanine) and Y (tyrosine).

[0053] If the amino acid sequence of the peptides at position X7 contained an amino acid from the group of R and L, particularly preferably R, the substantivity of the peptides to human hair was particularly high.

[0054] For this reason, a peptide according to the invention is characterized in a further particularly preferred embodiment in that

[0055] (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (X a )nX5X6X7X8X9(Xb)m where

[0056] X7 is an amino acid selected from the group consisting of R and L, particularly preferably R.

[0057] X8 is an amino acid selected from the group consisting of K (lysine), T (threonine), N (asparagine), A (alanine), L (leucine), F (phenylalanine), D (aspartic acid), R (arginine), and G (glycine). Particularly good results were obtained when X8 is an amino acid selected from the group consisting of K, T, N, and F, preferably from the group consisting of K, T, and N, particularly preferably from the group consisting of K and T.

[0058] In a further particularly preferred embodiment, a peptide according to the invention is characterized in that

[0059] (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (X a )nX5X6X7X8X9(Xb)m where

[0060] X8 is an amino acid selected from the group consisting of K, T, N and F, preferably from the group consisting of K, T and N, particularly preferably from the group consisting of K and T.

[0061] At position X9 there is an amino acid selected from the group consisting of R (arginine), L (leucine), I (isoleucine), A (alanine), Q (glutamine), P (proline), H (histidine), G (glycine) and W (tryptophan). If X9 stands for an amino acid selected from the group consisting of R, L, A and P, preferably from the group consisting of R, L and P, particularly preferably from the group consisting of R and L, most preferably R, a particularly good durability on hair strands was observed for the peptides according to the invention.

[0062] In a further particularly preferred embodiment, a peptide according to the invention is characterized in that

[0063] (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (X a)nX5X6X7X8X9(Xb)m where

[0064] X9 is an amino acid selected from the group consisting of R, L, A and P, preferably from the group consisting of R, L, and P, particularly preferably from the group consisting of R and L, most preferably R.

[0065] The residue Xb stands independently for any amino acid, ie Xb stands, for example, for an amino acid from the group of glycine (G), alanine (A), valine (V), leucine (L), isoleucine (I), phenylalanine (F), serine (S), threonine (T), proline (P), methionine (M), cysteine ​​(C), histidine (H), lysine (K), arginine (R), glutamine (Q), asparagine (N), aspartic acid (D), glutamic acid (E), tyrosine (Y) and tryptophan (W).

[0066] In a further embodiment, particularly preferred is a peptide comprising or consisting of an amino acid sequence of 5 to 50 amino acids, wherein

[0067] (a) the amino acid sequence in N- to C-terminal orientation has the following sequence, (Xa )nX5X6X7X8X9(Xb)m where each X a independently of one another is any amino acid, n is an integer from 0 to 45, preferably from 0 to 4,

[0068] X5 is an amino acid selected from the group consisting of P, N, and C,

[0069] Xe is an amino acid selected from the group consisting of T, R, M and P,

[0070] X7 is an amino acid selected from the group consisting of R and L

[0071] Xe is an amino acid selected from the group consisting of K, T, N and F,

[0072] X9 is an amino acid selected from the group consisting of R, L, A and P, each Xb is independently any amino acid, and m is an integer from 0 to 45, preferably from 0 to 7, more preferably from 0 to 3, with the proviso that when X5 is P, X9 does not simultaneously also represent P.

[0073] In a further embodiment, particularly preferred is a peptide comprising or consisting of an amino acid sequence of 5 to 50 amino acids, wherein

[0074] (a) the amino acid sequence in N- to C-terminal orientation has the following sequence, (X a )nX5X6X7X8X9(Xb)m where each X a independently of one another is any amino acid, n is an integer from 0 to 45, preferably from 0 to 4,

[0075] X5 stands for the amino acid P,

[0076] Xe is an amino acid selected from the group consisting of T and R,

[0077] X7 stands for the amino acid R,

[0078] Xe is an amino acid selected from the group consisting of K and T,

[0079] X9 is an amino acid selected from the group consisting of R and L, each Xb is independently any amino acid, and m is an integer from 0 to 45, preferably from 0 to 7, more preferably from 0 to 3.

[0080] The index number m stands for an integer from 0 to 45. Preferably, m stands for an integer from 0 to 7. Most preferably, m stands for an integer from 0 to 3. For example, if m stands for the number 3, this means that after the chain of amino acids X5X6X7X8X9 (i.e. towards the C-terminal end of the amino acid sequence) there are three further amino acids of your choice, so that the peptide then has the sequence (X a )nX5X6X7XsX9XbXbXb.

[0081] Furthermore, for the peptides according to the invention of embodiment (a), the proviso is that if X5 stands for P (proline), X9 does not simultaneously stand for P (proline).

[0082] All of the aforementioned features can be implemented individually or in any combination. However, certain combinations of the aforementioned radicals and index numbers are particularly well suited to solving the problem of the invention and are therefore particularly well suited.

[0083] Of the peptides investigated for this invention, those peptides that have shown particularly good adhesion to human hair comprise an amino acid sequence of 7 to 16 amino acids, with at least one further specific amino acid located both before and after the sequence X5X6X7X8X9. The particularly preferred peptides in embodiment (a) have the following sequence in N- to C-terminal orientation (X a )nX4X5X6X7X8X9Xl0(Xb)m

[0084] This means that the amino acid at position X4 is no longer just any amino acid (Xa), but rather an amino acid X4 selected from the group consisting of R (arginine), S (serine), I (isoleucine), V (valine), H (histidine), F (phenylalanine), T (threonine), G (glycine), L (leucine), E (glutamic acid), Q (glutamine) and K (lysine).

[0085] Furthermore, the amino acid at position X10 no longer represents any amino acid (Xb), but rather an amino acid X10 selected from the group consisting of I (isoleucine), K (lysine), S (serine), Q (glutamine), R (arginine), L (leucine), H (histidine), A (alanine), and F (phenylalanine). In this context, n preferably represents the number 0 to 3, particularly preferably the number 3, and m represents the number 0 to 6, particularly preferably the number 0 to 2, most preferably the number 2.

[0086] In a further particularly preferred embodiment, a peptide according to the invention in embodiment (a) is characterized in that

[0087] (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (X a )nX4X5X6X7X8X9Xl0(Xb)m where n stands for the number 0 to 3, preferably for the number 3,

[0088] X4 is an amino acid selected from the group consisting of R, S, I, V, H, F, T, G, L, E, Q and K,

[0089] X10 is an amino acid selected from the group consisting of I, K, S, Q, R, L, H, A and F, and m is the number 0 to 6, preferably the number 0 to 2, particularly preferably the number 2.

[0090] The substantivity to human hair could be further increased when X4 is an amino acid selected from the group consisting of R, S, I, F and G, preferably from the group consisting of R, S, I and F, particularly preferably from the group consisting of R, S and I. Likewise, positive effects with regard to the adhesion to human hair were achieved when X10 was an amino acid selected from the group consisting of I, K and R, preferably from the group consisting of I and K, particularly preferably I.

[0091] In a further particularly preferred embodiment, a peptide according to the invention in embodiment (a) is characterized in that

[0092] (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (X a )nX4X5X6X7X8X9Xl0(Xb)m where

[0093] X4 is an amino acid selected from the group consisting of R, S, I, F and G, preferably from the group consisting of R, S, I and F, particularly preferably from the group consisting of R, S and I, and / or

[0094] X10 is an amino acid selected from the group consisting of I, K and R, preferably from the group consisting of I and K, particularly preferably I.

[0095] Of the series of peptides investigated for this invention, those peptides that have demonstrated even better adhesion to human hair include an amino acid sequence of 9 to 16 amino acids, with at least one other specific amino acid located both before and after the sequence X4X5X6X7X8X9X10. The particularly preferred peptides in embodiment (a) have the following sequence in N- to C-terminal orientation: (Xa) 0X3X4X5X6X7X8X9X10X11 (Xb)m.

[0096] This means that the amino acid at position X3 is no longer just any amino acid (Xa), but rather an amino acid X3 selected from the group consisting of P (proline), R (arginine), I (isoleucine), N (asparagine), C (cysteine), L (leucine), A (alanine), V (valine) and Q (glutamine).

[0097] Furthermore, the amino acid at position X11 no longer stands for any amino acid (Xb), but for an amino acid X11, which is selected from the group consisting of Xn is an amino acid selected from the group consisting of K (lysine), R (arginine), I (isoleucine), N (asparagine), G (glycine), E (glutamic acid), V (valine), A (alanine), S (serine) and W (tryptophan). In this context, n preferably stands for the number n stands for the number 0 to 2, preferably for the number 2, and m stands for the number 0 to 5, preferably for the number 0 or 1, particularly preferably for the number 1.

[0098] In a further particularly preferred embodiment, a peptide according to the invention in embodiment (a) is characterized in that

[0099] (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (Xa) 0X3X4X5X6X7X3X9X10X11 (Xb)m where n is the number 0 to 2, preferably the number 2,

[0100] X3 is an amino acid selected from the group consisting of P, R, I, N, C, L, A, V and Q,

[0101] X11 is an amino acid selected from the group consisting of K, R, I, N, G, E, V, A, S and W, and m is the number 0 to 5, preferably the number 0 or 1, particularly preferably the number 1.

[0102] Particularly positive effects with regard to substantivity to the hair fiber were observed when X3 was an amino acid selected from the group consisting of P, R and N, preferably from the group consisting of P and R, most preferably P.

[0103] Also positive effects on adhesion to human hair were found when Xu was an amino acid selected from the group consisting of K and R, most preferably K.

[0104] In a further particularly preferred embodiment, a peptide according to the invention is characterized in that

[0105] (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (Xa) 01X3X4X5X6X7X3X9X10X11 (Xb)m where

[0106] X3 is an amino acid selected from the group consisting of P, R and N, preferably from the group consisting of P and R, most preferably P, and / or

[0107] X11 is an amino acid selected from the group consisting of K and R, most preferably K.

[0108] The very best results were obtained with peptides comprising an amino acid sequence of 12 amino acids, where the amino acid sequence (a) has the sequence X1X2X3X4X5X6X7X8X9X10X11X12 in the N- to C-terminal orientation. In this case, the sequence X5X6X7X8X9 is preceded (i.e., toward the N-terminal end) by not four random amino acids, but by the defined amino acids X1X2X3X4. X3 and X, as well as their preferred and particularly preferred embodiments, have already been defined.

[0109] Xi is an amino acid selected from the group consisting of Q (glutamine), R (arginine), H (histidine), N (asparagine), I (isoleucine), T (threonine), A (alanine), D (aspartic acid), and G (glycine). X2 is an amino acid selected from the group consisting of H (histidine), N (asparagine), M (methionine), S (serine), E (glutamic acid), W (tryptophan), P (proline), V (valine), A (alanine), and F (phenylalanine).

[0110] Beyond (i.e., toward the C-terminal end) of the sequence X5X6X7X8X9, the peptide no longer contains any three amino acids, but rather the defined amino acids X10X11X12. X10 and Xu, as well as their preferred and particularly preferred embodiments, have already been defined.

[0111] X12 is an amino acid selected from the group consisting of P (proline), R (arginine), Q (glutamine), I (isoleucine), T (threonine), E (glutamic acid), N (asparagine), L (leucine), A (alanine) and G (glycine).

[0112] In a further particularly preferred embodiment, a peptide according to the invention is characterized in that

[0113] (a) the amino acid sequence in N- to C-terminal orientation has the following sequence

[0114] X1X2X3X4X5X6X7X8X9X10X11X12 where

[0115] Xi is an amino acid selected from the group consisting of Q, R, H, N, I, T, AD and G, and

[0116] X2 is an amino acid selected from the group consisting of H, N, M, S, E, W, P, V, A and F, and

[0117] X12 is an amino acid selected from the group consisting of P, R, Q, I, T, E, N, L, A and G.

[0118] Particularly positive effects with regard to substantivity to the hair fiber were observed when Xi was an amino acid selected from the group consisting of Q, R, I and A, preferably from the group consisting of Q, R and I, particularly preferably from the group consisting of Q and I.

[0119] It also had positive effects with regard to adhesion to human hair when X2 is an amino acid selected from the group consisting of H, N, S and P, preferably from the group consisting of H, N and S, particularly preferably from the group consisting of H and N.

[0120] Furthermore, good adhesion to the hair fiber was observed when X12 is an amino acid selected from the group consisting of P, R, Q and T, preferably from the group consisting of P, R and Q, particularly preferably from the group consisting of P and R.

[0121] In a further particularly preferred embodiment, a peptide according to the invention is characterized in that (a) the amino acid sequence in N- to C-terminal orientation has the following sequence X1X2X3X4X5X6X7X8X9X10X11X12 where

[0122] Xi is an amino acid selected from the group consisting of Q, R, I and A, preferably from the group consisting of Q, R and I, particularly preferably from the group consisting of Q and I, and / or

[0123] X2 is an amino acid selected from the group consisting of H, N, S and P, preferably from the group consisting of H, N and S, particularly preferably from the group consisting of H and N, and / or

[0124] X12 is an amino acid selected from the group consisting of P, R, Q and T, preferably from the group consisting of P, R and Q, particularly preferably from the group consisting of P and R.

[0125] In the context of embodiment (b), the peptide according to the invention comprises a

[0126] Amino acid sequence of 5 to 50 amino acids, or it consists of this amino acid sequence, wherein the amino acid sequence has at least 85%, preferably at least 86%, more preferably at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with one of the amino acid sequences mentioned in SEQ ID NOs: 1 - 17.

[0127] The peptide according to the invention preferably comprises or consists of an amino acid sequence which has at least 85%, preferably at least 86%, more preferably at least 87%, even more preferably at least 88%, even more preferably at least 89%, even more preferably at least 90%, even more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, even more preferably at least 99%, in particular 100% sequence identity with one of the following amino acid sequences: QHPRPTRKRIKP (SEQ ID No: 1), QNRSPRRTRKRR (SEQ ID No: 2), QNRIPTRTRKKQ (SEQ ID No: 3), RNPIPMRNLKRQ (SEQ ID No: 4), HHRRHGRTIIIR (SEQ ID No: 5), HMISTMNAASRR (SEQ ID No: 6), RSIVTFSLRQNRC (SEQ ID No: 7), NERHNRRLQGRI (SEQ ID No: 8),ISNFNPRFPRRT (SEQ ID No: 9), TWRTRNLKISEE (SEQ ID No: 10), APNGCRLNAKRR (SEQ ID No: 11), HVCLRSIDHSVN (SEQ ID No: 12), RALRALQALQALEALC (SEQ ID No: 13), RALEALWRALEALC (SEQ ID No: 14), DHAQRYGAGHSG (SEQ ID No: 15), HFVKTPARWAWG (SEQ ID No: 16) and / or GHQGHWYGMFRA (SEQ ID No: 17).

[0128] The peptides with the amino acid sequences of SEQ ID NOs: 1-17 have demonstrated very good durability even on hair of different types and ethnic origins. However, particularly good results were obtained with certain amino acid sequences. The peptides according to one of SEQ ID NOs: 1-11, and even more preferably according to one of SEQ ID NOs: 1-4, 9, and 11, have proven particularly suitable. The very best results were obtained with SEQ ID NOs: 1 and / or SEQ ID NOs: 2.

[0129] In a further very particularly preferred embodiment, a peptide according to the invention is characterized in that the peptide has the amino acid sequence according to one of SEQ ID NOs: 1-11, preferably according to one of SEQ ID NOs: 1-4, 9 and 11, which has at least 85%, preferably at least 90%, more preferably at least 95% and very particularly preferably 100% sequence identity to the specified sequences.

[0130] Determination of nucleic acid or amino acid sequences

[0131] The identity of nucleic acid or amino acid sequences can be determined, for example, by sequence comparison. This sequence comparison is based on the state-of-the-art and commonly used BLAST algorithm (see, for example, Altschul, SF, Gish, W., Miller, W., Myers, EW & Lipman, DJ (1990) "Basic local alignment search tool." J. Mol. Biol. 215:403-410, and Altschul, Stephan F., Thomas L. Madden, Alejandro A. Schaffer, Jinghui Zhang, Hheng Zhang, Webb Miller, and David J. Lipman (1997): "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs"; Nucleic Acids Res., 25, pp. 3389-3402) and is essentially performed by matching similar sequences of nucleotides or amino acids in the nucleic acid or amino acid sequences. A tabular assignment of the relevant positions is called alignment.Another state-of-the-art algorithm is the FASTA algorithm. Sequence alignments, especially multiple sequence alignments, are created using computer programs. Commonly used algorithms include the Clustal series (see, for example, Chenna et al. (2003): Multiple sequence alignment with the Clustal series of programs. Nucleic Acid Research 31, 3497-3500), T-Coffee (see, for example, Notredame et al. (2000): T-Coffee: A novel method for multiple sequence alignments. J. Mol. Biol. 302, 205-217), or programs based on these programs or algorithms. Furthermore, sequence comparisons (alignments) are possible using the computer program Vector NTIR Suite 10.3 (Invitrogen Corporation, 1600 Faraday Avenue, Carlsbad, California, USA) with the specified standard parameters, whose AlignX module for sequence comparisons is based on ClustalW.Unless otherwise stated, sequence identity reported herein is determined using the BLAST algorithm.

[0132] Such a comparison also allows a statement to be made about the similarity of the compared sequences. This is usually expressed as percent identity, i.e., the proportion of identical nucleotides or amino acid residues at the same or corresponding positions in an alignment. The broader term "homology" in amino acid sequences includes conserved amino acid substitutions, i.e., amino acids with similar chemical activity, since these usually exert similar chemical activities within the peptide / protein. Therefore, the similarity of the compared sequences can also be expressed as percent homology or percent similarity. Identity and / or homology statements can be made for entire peptides, polypeptides, or genes, or only for individual regions. Homologous or identical regions of different nucleic acid or amino acid sequences are therefore defined by similarities in the sequences.Such regions often exhibit identical functions. They can be small and comprise only a few nucleotides or amino acids. However, such small regions often perform essential functions for the overall activity of the peptide / protein. It may therefore be useful to refer sequence matches only to individual, possibly small regions. Unless otherwise stated, statements of identity or homology in this application refer to the entire length of the respective nucleic acid or amino acid sequence.

[0133] The peptide or protein concentration can be determined using known methods, for example the BCA method (bicinchoninic acid; 2,2'-biquinolyl-4,4'-dicarboxylic acid) or the biuret method (AG Gornall, CS Bardawill, and MM David, J. Biol. Chem., 177 (1948), pp. 751-766). Those skilled in the art of peptide and protein technology are familiar with a variety of suitable methods for determining peptide or protein concentration that can be applied within the scope of this invention.

[0134] Peptides according to the invention may contain amino acid modifications, particularly amino acid substitutions, insertions, or deletions. Such peptides are further developed, for example, through targeted genetic modification, i.e., through mutagenesis techniques, and optimized for specific applications or with regard to specific properties (e.g., with regard to their stability, binding, etc.).

[0135] For example, targeted mutations such as substitutions, insertions, or deletions can be introduced into known molecules to alter specific properties. This can be achieved by altering the surface charges and / or the isoelectric point of the molecules, and thus their interactions with a surface. For example, the net charge of the peptides can be altered to influence substrate binding. Alternatively or additionally, one or more corresponding mutations can increase, for example, the stability or adsorption of the peptide. Advantageous properties of individual mutations, e.g., individual substitutions, can complement each other.

[0136] The following convention is used to describe substitutions that affect exactly one amino acid position (amino acid exchanges): first, the naturally occurring amino acid is named using the internationally used one-letter code, followed by the corresponding sequence position, and finally the inserted amino acid. Multiple exchanges within the same peptide chain are separated by slashes. In the case of insertions, additional amino acids are named after the sequence position. In the case of deletions, the missing amino acid is replaced by a symbol, such as an asterisk or a dash, or an A is indicated in front of the corresponding position. For example, P9T describes the substitution of proline at position 9 with threonine, P9TH the insertion of histidine after the amino acid threonine at position 9, and P9* or AP9 the deletion of proline at position 9.This nomenclature is known to those skilled in the field of enzyme technology.

[0137] Thus, the invention also encompasses peptides which are characterized in that they are obtainable from a peptide as described above as the starting molecule, for example from a molecule having one of the amino acid sequences according to SEQ ID NOs: 1-17, preferably according to one of SEQ ID NOs 1-11, particularly preferably according to one of SEQ ID NOs: 1-4, 9 and 11, on which, for example, one or more amino acid substitutions, including single or multiple conservative amino acid substitutions, have been carried out, wherein the resulting peptide has at least 85% sequence identity with one of the amino acid sequences according to SEQ ID NOs: 1-17, preferably 1-11, particularly preferably 1-4, 9 and 11. The term "conservative amino acid substitution" means the exchange (substitution) of one amino acid residue for another amino acid residue, wherein this exchange does not lead to a change in the polarity or charge at the position of the exchanged amino acid, e.g.the exchange of one nonpolar amino acid residue for another nonpolar amino acid residue. Conservative amino acid substitutions within the scope of the invention include, for example: G=A=S, I=V=L=M, D=E, N=Q, K=R, Y=F, S=T, G=A=I=V=L=M=Y=F=W=P=S=T. However, it may be preferred that such exchanges do not have glycine or tyrosine as the target amino acid, or, for example, an amino acid that has a low alpha-helix-forming potential.

[0138] In preferred embodiments, the peptide of the invention can also be modified. Preferred modifications can include, for example, coupling the peptide of the invention with certain other molecules or chemical groups, for example, organic (macro)molecules, e.g., via a covalent bond or a linker via a suitable amino acid of the chain and / or N- and / or C-terminal.

[0139] If the peptide of the invention is coupled to at least one other (macro)molecule, it can also be referred to as a peptide derivative. The peptide of the invention is then derivatized. In such embodiments, the peptide can act as an adhesion tag, causing the binding of a molecule coupled to it to the desired surface. Such molecules can also be referred to as conjugates.

[0140] In some embodiments, the said peptides according to the invention, which may, for example, have the amino acid cysteine ​​N- or C-terminally for coupling purposes, are coupled with biotin (functionally modified), preferably at a suitable amino acid of the chain and / or N- and / or C-terminally.

[0141] Production of the peptides

[0142] The peptides according to the invention can be prepared chemically by known methods of peptide synthesis, for example by solid-phase synthesis according to Merrifield.

[0143] However, it is preferred to produce the peptides according to the invention using recombinant processes. According to the invention, this refers to all genetic engineering or microbiological processes based on the introduction of the genes for the peptides of interest into a host organism suitable for production and their transcribing and translating by this host organism (collectively referred to as biotechnological processes within the scope of this invention). For example, the genes in question are introduced via vectors, in particular expression vectors; but also via those that allow the gene of interest to be inserted into an already existing genetic element in the host organism, such as the chromosome or other vectors. The functional unit consisting of gene and promoter, and possibly other genetic elements, is typically referred to as an expression cassette. However, it does not necessarily have to be present as a physical unit.

[0144] The peptides according to the invention are particularly preferably produced as polypeptides (multimers) and subsequently cleaved into the functional peptides. Very particularly preferred multimers have 2 to 10 peptide units (each according to the invention), which are each separated from one another by spacers of 0 to 4 amino acids in length (for example 1, 2, 3 or 4 amino acids). These spacers can consist, for example, of the amino acids glycine (G), alanine (A) and serine (S). Alternatively, the spacers can also be or comprise cleavage sites for specific proteases / peptidases, in particular endopeptidases, or can form such cleavage sites together with parts of the peptide according to the invention.

[0145] Using methods that are generally known today, such as chemical synthesis or the polymerase chain reaction (PCR) in conjunction with standard molecular biological and / or protein chemical methods, it is possible for a person skilled in the art to produce the corresponding nucleic acids up to complete genes based on known DNA and / or amino acid sequences and then to use these for the synthesis of peptides and polypeptides in suitable host cells.

[0146] In particularly preferred embodiments, the peptide is produced by biotechnological processes as defined above.

[0147] For the purposes of the present invention, vectors are understood to be elements consisting of nucleic acids that contain a gene of interest as a characteristic nucleic acid region. They are capable of establishing this gene in a species or cell line over several generations or cell divisions as a stable genetic element that replicates independently of the rest of the genome. Vectors, particularly when used in bacteria, are special plasmids, i.e., circular genetic elements. In genetic engineering, a distinction is made between vectors that serve for storage and thus, to a certain extent, also for genetic engineering work, the so-called cloning vectors, and those that fulfill the function of implementing the gene of interest in the host cell, i.e., enabling the expression of the respective peptide. These vectors are referred to as expression vectors.

[0148] The nucleic acid encoding a peptide according to the invention or a multimer of such a peptide, which constitutes a further aspect of the invention, can, for example, be cloned into a vector. The molecular biological dimension of the invention thus consists in vectors containing the genes for the corresponding peptides. These can include, for example, vectors derived from bacterial plasmids, viruses, or bacteriophages, or predominantly synthetic vectors or plasmids with elements of various origins. With the additional genetic elements present, vectors are capable of establishing themselves as stable units in the respective host cells over several generations. It is irrelevant for the purposes of the invention whether they establish themselves extrachromosomally as independent units or integrate into a chromosome. Which of the numerous systems known from the prior art is chosen depends on the individual case.The decisive factors may include, for example, the achievable copy number, the available selection systems, including in particular antibiotic resistance, or the cultivability of the host cells capable of taking up the vectors.

[0149] The vectors provide suitable starting points for molecular biological and biochemical studies of the gene in question or associated peptide, for further developments according to the invention, and ultimately for the amplification and production of peptides according to the invention. They represent further aspects of the present invention.

[0150] Preferred embodiments of the present invention are cloning vectors. In addition to storage, biological amplification, or selection of the gene of interest, these vectors are suitable for characterizing the gene in question, for example, by creating a restriction map or sequencing. Cloning vectors are also preferred embodiments of the present invention because they represent a transportable and storable form of the claimed DNA. They are also preferred starting points for molecular biological techniques that are not dependent on cells, such as the polymerase chain reaction.

[0151] Expression vectors are chemically similar to cloning vectors, but differ in those partial sequences that enable them to replicate in host organisms optimized for peptide production and to express the contained gene there. Preferred embodiments are expression vectors that themselves carry the genetic elements necessary for expression. Expression is influenced, for example, by promoters that regulate gene transcription. Thus, expression can occur through the natural promoter originally located upstream of this gene, but also after genetic fusion, through a promoter of the host cell provided on the expression vector, or through a modified or completely different promoter from another organism.

[0152] Preferred embodiments are expression vectors that can be regulated by changes in culture conditions or the addition of specific compounds, such as cell density or specific factors. Embodiments of the present invention can also be cell-free expression systems in which peptide biosynthesis is reproduced in vitro. Such expression systems are also established in the prior art.

[0153] The in vivo synthesis of a peptide according to the invention, i.e., by living cells, requires the transfer of the corresponding gene into a host cell, the so-called transformation. In principle, all organisms are suitable as host cells, i.e., prokaryotes, eukaryotes, or cyanophyta. Preferred host cells are those that are genetically easy to handle, for example, with regard to transformation with the expression vector and its stable establishment, for example, unicellular fungi such as yeasts or bacteria. Furthermore, preferred host cells are characterized by good microbiological and biotechnological handling. This applies, for example, to easy culturability, high growth rates, low requirements for fermentation media, and good production and secretion rates for foreign peptides.Often, the optimal expression systems for a specific case must be determined experimentally from the wealth of different systems available according to the state of the art. Each peptide of the invention can be obtained from a variety of host organisms in this way.

[0154] A further aspect of the present invention relates to such host cells. Preferred embodiments are host cells whose activity can be regulated by genetic regulatory elements, which are provided, for example, on the expression vector, but can also be present in these cells from the outset. These cells can be stimulated to express, for example, by the controlled addition of chemical compounds that serve as activators, by changing the cultivation conditions, or upon reaching a certain cell density. This enables very economical production of the peptides of interest.

[0155] Preferred host cells are prokaryotic or bacterial cells. Bacteria are generally characterized by shorter generation times and lower demands on cultivation conditions compared to eukaryotes. This allows for the establishment of cost-effective methods for obtaining peptides according to the invention. In Gram-negative bacteria, such as E. coli, a large number of peptides are secreted into the periplasmic space, i.e., the compartment between the two membranes enclosing the cells. This can be advantageous for specific applications. Gram-positive bacteria, such as Bacilli or Actinomycetes or other members of the Actinomycetales, in contrast, do not have an outer membrane, so secreted peptides are immediately released into the nutrient medium surrounding the cells, from which, according to another preferred embodiment, the expressed peptides according to the invention can be directly purified.A variation of this principle is represented by expression systems in which additional genes, for example those provided on other vectors, influence the production of peptides of the invention. These can be modifying gene products or those that are to be co-purified with the peptide of the invention.

[0156] Due to the extensive experience gained, for example, with regard to molecular biological methods and culturability with coliform bacteria, these represent preferred embodiments of the present invention. Particular preference is given to those of the genera Escherichia coli, in particular non-pathogenic strains suitable for biotechnological production.

[0157] Representative representatives of these genera are the K12 derivatives and the B strains of Escherichia coli. Strains that can be derived from these using known genetic and / or microbiological methods and can thus be considered their derivatives. These are of the greatest importance for genetic and microbiological work and are preferably used for the development of the methods according to the invention. Such derivatives can, for example, be modified by deletion or insertion mutagenesis with regard to their culture requirements, have different or additional selection markers, or express different or additional peptides. In particular, these can be derivatives that express other commercially interesting peptides in addition to the peptide produced according to the invention.

[0158] Also preferred are microorganisms characterized by being obtained after transformation with one of the vectors described above. These can be, for example, cloning vectors that have been introduced into any bacterial strain for storage and / or modification. Such steps are common in the storage and further development of relevant genetic elements. Since the relevant genetic elements can be directly transferred from these microorganisms into Gram-negative bacteria suitable for expression, the preceding transformation products also represent realizations of the relevant subject matter of the invention.

[0159] Eukaryotic cells can also be suitable for producing peptides according to the invention. Examples include fungi such as Actinomycetes or yeasts such as Saccharomyces or Kluyveromyces. This can be particularly advantageous, for example, if the peptides are to undergo specific modifications during their synthesis that enable such systems. These include, for example, the binding of low-molecular-weight compounds such as membrane anchors or oligosaccharides. In the context of this invention, this would be an example of a functionally modified peptide. The host cells of the process according to the invention are cultivated and fermented in a conventional manner, for example, in discontinuous or continuous systems. In the first case, a suitable nutrient medium is inoculated with the recombinant bacterial strains, and the product is harvested from the medium after a period to be determined experimentally.Continuous fermentations are characterized by reaching a steady state in which cells partially die but also regrow over a comparatively long period of time and at the same time product can be removed from the medium.

[0160] Fermentation processes are well known in the art and represent the actual large-scale production step; followed by a suitable purification method.

[0161] All fermentation processes based on one of the above-described processes for producing the recombinant peptides represent correspondingly preferred embodiments of this subject matter of the invention.

[0162] In this case, the optimal conditions for the production processes used, for the host cells and / or the peptides to be produced must be determined experimentally on the basis of the previously optimized culture conditions of the strains in question according to the knowledge of the person skilled in the art, for example with regard to fermentation volume, media composition, oxygen supply or stirrer speed.

[0163] Fermentation processes characterized by a feed-in strategy are also possible. In this case, the media components consumed by the ongoing cultivation are added; this is also referred to as a supplementary feeding strategy. This can achieve significant increases in both cell density and dry biomass, and / or, above all, the activity of the peptide of interest.

[0164] Analogously, the fermentation can also be designed in such a way that undesirable metabolic products are filtered out or neutralized by adding buffer or appropriate counterions.

[0165] The produced peptide can subsequently be harvested from the fermentation medium. This fermentation process is preferred over product preparation from dry matter, but requires the provision of suitable secretion markers and transport systems. Without secretion, purification of the peptide from the cell mass may be necessary. Various methods are known for this, such as precipitation, e.g., with ammonium sulfate or ethanol, or chromatographic purification, if necessary to homogeneity. However, the majority of the described technical processes should be able to operate with an enriched, stabilized preparation.

[0166] All of the elements already outlined above can be combined into processes for producing peptides according to the invention. These processes for producing the peptides according to the invention represent further aspects of the present invention. A multitude of possible combinations of process steps are conceivable for each peptide according to the invention. Optimal conditions can be determined experimentally for each specific case.

[0167] Nucleic acid sequences

[0168] A further aspect of the present invention are (isolated) nucleic acid sequences which encode a peptide according to the invention or which encode a peptide comprising or consisting of an amino acid sequence according to one of SEQ ID NOs: 1-17, preferably 1-11, particularly preferably 1-4, 9 and 11, which have at least 85%, preferably at least 86%, more preferably at least 87%, more preferably at least 88%, even more preferably at least 89%, even more preferably at least 90%, even more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, even more preferably at least 99% and in particular 100% sequence identity with the said sequences.

[0169] A second subject matter of the present invention is a nucleic acid sequence coding for a peptide according to one of claims 1 to 13, preferably for a peptide comprising or consisting of an amino acid sequence according to one of SEQ ID NOs: 1-17.

[0170] Agents for treating keratin fibers

[0171] A further subject of the present invention is an agent for treating keratin fibers, in particular human hair, containing in a cosmetic carrier at least one peptide as disclosed in detail in the description of the first subject of the invention.

[0172] Keratin fibers, keratin-containing fibers, or keratin fibers are understood to mean fur, wool, feathers, and especially human hair. Although the agents according to the invention are primarily suitable for treating keratin fibers, especially human hair, in principle there is nothing to prevent their use in other areas as well.

[0173] The agent contains the peptide(s) in a cosmetic carrier, preferably in a suitable aqueous, alcoholic, or aqueous-alcoholic carrier. For hair treatment, such carriers can be, for example, creams, emulsions, gels, or surfactant-containing foaming solutions, such as shampoos, foam aerosols, foam formulations, or other preparations suitable for application to the hair. Agents for treating keratin fibers, in particular human hair, are particularly preferably creams or emulsions.

[0174] In a preferred embodiment, the cosmetic carrier is a water-containing cosmetic carrier. In a further preferred embodiment, the cosmetic carrier is a carrier containing water and ethanol.

[0175] In a further preferred embodiment, the cosmetic carrier is a carrier which contains water and a preservative, preferably a preservative from the group consisting of sodium sulfite, potassium sulfite, potassium hydrogen sulfite, sodium hydrogen sulfite, zinc pyrithione, benzoic acid, salicylic acid, sorbic acid, formic acid, propionic acid and the physiologically acceptable salts of the aforementioned acids, 2-phenoxyethanol, benzyl alcohol and 1-phenoxypropan-2-ol.

[0176] Depending on the desired coverage of the surface, the peptide(s) according to the invention can be present in the product in various amounts. The product preferably contains—based on the total weight of the product—one or more peptides according to the invention in a total amount of 0.0001 to 20 wt.%, preferably 0.01 to 15 wt.%, more preferably 0.1 to 10 wt.%, and most preferably 0.2 to 7.5 wt.%.

[0177] Methods for treating keratin fibers

[0178] A further subject of the present invention is a method for treating keratin fibers, in particular human hair, wherein a cosmetic agent containing at least one peptide, as disclosed in detail in the description of the first subject of the invention, is applied to the keratin fibers and optionally rinsed out again after a contact time.

[0179] The contact time of the agent on the keratin fibers or hair can be, for example, 1 to 60 minutes, preferably 15 to 55 minutes, and most preferably 30 to 50 minutes. After this period, the agent can be rinsed out of the keratin fibers with water or with water and shampoo. In another embodiment, however, a leave-on application of the agent is conceivable, in which the keratin fibers still coated with the agent are dried.

[0180] For the further embodiments of the agent and method according to the invention, what has been said regarding the peptides according to the invention applies mutatis mutandis.

[0181] Examples

[0182] 1. peptides used and their peptide sequences

[0183] Table 1

[0184] Some of the peptides originate from E. coli display database screenings (e.g. P8C), other peptides are either further developments of these or pure designer peptides (e.g. RAL2QEC and TF12 (RALF-W)).

[0185] Negative control

[0186] Table 2

[0187] The peptides used as negative controls are peptides not according to the invention that have no or almost no adhesion to the hair.

[0188] 2. Investigation of the adhesion of the synthesized peptides with regard to their substrate specificity on different hair types

[0189] The hair strands of different types were washed, rinsed and dried with a 10% aqueous sodium lauryl sulfate solution, which was brought to a pH of 10 with lactic acid.

[0190] The hair was then cut into small pieces, and 10 to 11 mg were weighed out in each portion.

[0191] Each of the peptides was dissolved in water (0.01–0.3 mg / ml). A 300 μl (microliter) portion of this solution was added to each portion of cut hair. This mixture was incubated for one hour at room temperature with continuous shaking in a thermomixer (700 rpm).

[0192] 50 pl (microliters) of the supernatant peptide solution were pipetted into a microtiter plate, and this solution was spiked with 15 pl micrograms of fluorescamine.

[0193] Fluorescamine is the organic spiro compound 4-phenylspiro-[furan-2(3H),1-phthalan]-3,3'-dione (CAS No. 38183-12-9), which is used in protein analysis for the fluorescent labeling of amino acids and peptides. A free amino group of the protein is derivatized with fluorescamine. Fluorescamine does not exhibit fluorescence without derivatization; the derivative can be excited at 360 nm and emits at 465 nm.

[0194] Fluorescence measurements were then performed at 465 nm. After appropriate calibration, the concentration of the remaining peptide in the supernatant peptide solution can be calculated from the intensity of the fluorescence signal. The more peptides bind to the hair surface and are thus removed from the supernatant peptide solution, the lower the fluorescence signal in the solution. Accordingly, the higher the fluorescence signal, the poorer the adhesion of the peptides to the hair surface.

[0195] The adhesion of the peptides to the hair fiber, expressed as a percentage, was calculated using the following formula: conc.(Peptide)Start = Concentration of the peptide used in the aqueous solution conc.(Peptide)End = Concentration of the peptide in the aqueous solution of the supernatant after incubation

[0196] Four different hair types were examined in the measurements: Kerling hair type 3, Kerling hair type 9, Indian hair (root area) and Indian hair (tip area).

[0197] Table 3: Adhesion values ​​of the tested peptides on the four hair types examined

[0198] Comparison - negative control

[0199] Particularly good results were obtained with peptides G06 (No. 1), BioQR12 (No. 2), ConsS5 (No. 3), A09S5 (No. 4), BP2 (No. 9), and BP4 (No. 11). These peptides showed very good adhesion to hair of various types, and at least one hair type measured an adhesion of over 90%.

[0200] Within this list, peptides G06 (No. 1), BioQR12 (No. 2), ConsS5 (No. 3), and A09S5 (No. 4) showed even better performance.

[0201] The peptides G06 (No. 1) and BioQR12 (No. 2) showed the very best substantivity on all hair types, as they not only adhered well to Kerling-type hair, but also had a very high substantivity on Indian hair.

[0202] The peptides PepE (No. 18), AL-S1 (No. 19), and MS-S1 (No. 20) showed no significant adhesion to either Kerling 3 or Kerling 9 hair. Therefore, measurements on Indian hair were not performed for these negative controls.

Claims

Patent claims 1 . Peptide comprising or consisting of an amino acid sequence of 5 to 50 amino acids, wherein (a) the amino acid sequence in N- to C-terminal orientation has the following sequence, (X a )nX5X6X7X8X9(Xb)m where each X a independently of one another is any amino acid, n is an integer from 0 to 45, preferably from 0 to 4, X5 is an amino acid selected from the group consisting of P, H, T, N, R, C and A, Xe is an amino acid selected from the group consisting of T, R, M, G, F, P, N, S, L, Y and W, X7 is an amino acid selected from the group consisting of R, N, S, L, I, Q, W, G, A and Y, Xe is an amino acid selected from the group consisting of K, T, N, A, L, F, D, R and G, X9 is an amino acid selected from the group consisting of R, L, I, A, Q, P, H, G, W and M, each Xb is independently any amino acid, and m is an integer from 0 to 45, preferably from 0 to 7, particularly preferably from 0 to 3, with the proviso that when X5 stands for P, X9 does not simultaneously stand for P, and / or b) the amino acid sequence has at least 85%, preferably at least 86%, more preferably at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with one of the amino acid sequences mentioned in SEQ ID NOs: 1 - 17.

2. Peptide according to claim 1, characterized in that (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (Xa)nX5X6X7X8X9(Xb)m where X5 is an amino acid selected from the group consisting of P, N, and C, particularly preferably P.

3. Peptide according to one of claims 1 to 2, characterized in that (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (Xa)nX5X6X7X8X9(Xb)m where Xe is an amino acid selected from the group consisting of T, R, M and P, preferably from the group consisting of T, R and M, particularly preferably from the group consisting of T and R.

4. Peptide according to one of claims 1 to 3, characterized in that (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (X a )nX5X6X7X8X9(Xb)m where X7 is an amino acid selected from the group consisting of R and L, particularly preferably R.

5. Peptide according to one of claims 1 to 4, characterized in that (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (X a )nX5X6X7X8X9(Xb)m where X8 is an amino acid selected from the group consisting of K, T, N and F, preferably from the group consisting of K, T and N, particularly preferably from the group consisting of K and T.

6. Peptide according to one of claims 1 to 5, characterized in that (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (X a )nX5X6X7X8X9(Xb)m where X9 is an amino acid selected from the group consisting of R, L, A and P, preferably from the group consisting of R, L and P, particularly preferably from the group consisting of R and L, most preferably R.

7. Peptide according to any one of claims 1 to 6, comprising or consisting of an amino acid sequence of 7 to 16 amino acids, wherein (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (Xa )nX4X5X6X7X8X9Xl0(Xb)m where n stands for the number 0 to 3, preferably for the number 3, X4 is an amino acid selected from the group consisting of R, S, I, V, H, F, T, G, L, E, Q and K, X10 is an amino acid selected from the group consisting of I, K, S, Q, R, L, H, A and F, and m stands for the number 0 to 6, preferably the number 0 to 2, particularly preferably the number 2.

8. Peptide according to claim 7, characterized in that (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (X a )nX4X5X6X7X8X9Xl0(Xb)m where X4 is an amino acid selected from the group consisting of R, S, I, F and G, preferably from the group consisting of R, S, I and F, particularly preferably from the group consisting of R, S and I, and / or X10 is an amino acid selected from the group consisting of I, K and R, preferably from the group consisting of I and K, particularly preferably I.

9. Peptide according to any one of claims 1 to 8, comprising or consisting of an amino acid sequence of 9 to 16 amino acids, wherein (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (Xa) 0X3X4X5X6X7X3X9X10X11 (Xb)m where n is the number 0 to 2, preferably the number 2, X3 is an amino acid selected from the group consisting of P, R, I, N, C, L, A, V and Q, X11 is an amino acid selected from the group consisting of K, R, I, N, G, E, V, A, S and W, and m is the number 0 to 5, preferably the number 0 or 1, particularly preferably the number 1.

10. Peptide according to claim 9, characterized in that (a) the amino acid sequence in N- to C-terminal orientation has the following sequence (Xa) 01X3X4X5X0X7X8X9X10X11 (Xb)m where X3 is an amino acid selected from the group consisting of P, R and N, preferably from the group consisting of P and R, most preferably P, and / or X11 is an amino acid selected from the group consisting of K and R, most preferably K.

11. Peptide according to any one of claims 1 to 10, comprising or consisting of an amino acid sequence of 12 amino acids, wherein (a) the amino acid sequence in N- to C-terminal orientation has the following sequence X1X2X3X4X5X6X7X8X9X10X11X12 where Xi is an amino acid selected from the group consisting of Q, R, H, N, I, T, AD and G, and X2 is an amino acid selected from the group consisting of H, N, M, S, E, W, P, V, A and F, and X12 is an amino acid selected from the group consisting of P, R, Q, I, T, E, N, L, A and G.

12. Peptide according to claim 11, characterized in that (a) the amino acid sequence in N- to C-terminal orientation has the following sequence X1X2X3X4X5X6X7X8X9X10X11X12 where Xi is an amino acid selected from the group consisting of Q, R, I and A, preferably from the group consisting of Q, R and I, particularly preferably from the group consisting of Q and I, and / or X2 is an amino acid selected from the group consisting of H, N, S and P, preferably from the group consisting of H, N and S, particularly preferably from the group consisting of H and N, and / or X12 is an amino acid selected from the group consisting of P, R, Q and T, preferably from the group consisting of P, R and Q, particularly preferably from the group consisting of P and R.

13. Peptide according to one of claims 1 to 12, characterized in that the peptide has the amino acid sequence according to one of SEQ ID NOs: 1-11, preferably according to one of SEQ ID NOs: 1-4, 9 and 11, which have at least 85%, preferably at least 90%, more preferably at least 95% and most preferably 100% sequence identity to the specified sequences.

14. Nucleic acid sequence coding for a peptide according to any one of claims 1 to 13, preferably for a peptide comprising or consisting of an amino acid sequence according to any one of SEQ ID NOs: 1-17.

15. Agent for treating keratin fibers, in particular human hair, containing in a cosmetic carrier at least one peptide according to one of claims 1 to 13.

16. A method for treating keratin fibers, in particular human hair, wherein a cosmetic agent containing at least one peptide according to one of claims 1 to 13 is applied to the keratin fibers and optionally rinsed out again after a contact time.

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