METHOD FOR COUPLING KERATIN-BINDING POLYPEPTIDES WITH EFFECTOR MOLECULES WHICH SUPPORT CARBOXYLIC GROUPS OR SULFONIC ACID GROUPS

• US 20090156485A1
• Filed: 11/15/2006
• Published: 06/18/2009
• Est. Priority Date: 11/24/2005
• Status: Abandoned Application

• Current Assignee: BASF AG (BASF SE)
• Original Assignee: BASF SE
• Inventors: Barg, Heiko, Liebmann, Burghard, Ptock, Arne, Volkert, Martin, Reents, Heike
• Application Number:
  US12/094,803
• Publication Number:
  US 20090156485A1
• Time in Patent Office: Days
• US Class Current: 514/1.100
• CPC Class Codes: A61K 2800/57 Compounds covalently linked to a(n inert) carrier molecule, e.g. conjugates, pro-fragrances
• CPC Class Codes: A61K 2800/94 Involves covalent bonding to the substrate
• CPC Class Codes: A61K 47/62 the modifying agent being a protein, peptide or polyamino acid
• CPC Class Codes: A61K 47/6843 the antibody targeting a material from animals or humans
• CPC Class Codes: A61K 8/64 Proteins; Peptides; Derivatives or degradation products thereof
• CPC Class Codes: A61Q 1/02 Preparations containing skin colorants, e.g. pigments preparations in powder form A61Q1/12
• CPC Class Codes: A61Q 17/00 Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
• CPC Class Codes: A61Q 17/04 Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
• CPC Class Codes: A61Q 19/00 Preparations for care of the skin
• CPC Class Codes: A61Q 19/002 Aftershave preparations
• CPC Class Codes: A61Q 19/004 Aftersun preparations
• CPC Class Codes: A61Q 19/04 for chemically tanning the skin topical sun or radiation screening or tanning preparations A61Q17/04
• CPC Class Codes: A61Q 19/10 Washing or bathing preparations
• CPC Class Codes: A61Q 5/02 Preparations for cleaning the hair
• CPC Class Codes: A61Q 5/06 Preparations for styling the hair, e.g. by temporary shaping or colouring
• CPC Class Codes: A61Q 5/12 Preparations containing hair conditioners

Abstract

The invention relates to a method of producing keratin-binding effector molecules and to intermediates and end-products of the method according to the invention and to the use of the keratin-binding effector molecules produced according to the invention in dermocosmetics.

Claims

1. A method of producing a keratin-binding effector molecule comprising coupling an effector molecule (i) carrying at least one carboxyl or sulfonic acid group onto a keratin-binding polypeptide (ii) using a linker molecule (iii) which has at least two coupling functionalities which can enter into bonds chosen from the group consisting of amide, thioester, thioether, ester, sulfonic acid ester and sulfonamide bonds, and

(a) in a first coupling step, firstly binding the effector molecule (i) to the linker molecule (iii) via the carboxyl or sulfonic acid group by means of an ester or sulfonamide bond, and

(b) in another coupling step, coupling the reaction product from (a) to the keratin-binding polypeptide (ii) via a still free coupling functionality of the linker molecule (iii).

2. The method according to claim 1, where the coupling of the linker molecule (iii) with the effector molecule (i) described in (a) is a carbodiimide- or acid-chloride-mediated esterification reaction.

3. The method according to claim 1, where the effector molecule (i) is chosen from the group consisting of dyes, photoprotective agents, vitamins, provitamins, carotenoids, antioxidants and peroxide decomposers.

4. The method according to claim 1, where the keratin-binding polypeptide (ii) has a binding affinity to human skin, hair or nail keratin.

5. The method according to claim 1, where the keratin-binding polypeptide (ii) used

(a) comprises at least one of the sequences according to SEQ ID NO.:

2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 146, 150, 153, 156, 157, 158, 160, 162, 164, 166, 168 or 170, or

(b) corresponds to a polypeptide which is at least 40% identical to at least one of the sequences according to SEQ ID NO.;

2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 146, 150, 153, 156, 157, 158, 160, 162, 164, 166, 168 or 170 and is able to bind keratin.

6. The method according to claim 1, wherein the keratin-binding polypeptide (ii) used is encoded by a nucleic acid molecule comprising at least one nucleic acid molecule chosen from the group consisting of:

a) a nucleic acid molecule which encodes a polypeptide comprising the sequence shown in SEQ ID NO.;

2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 146, 150, 153, 156, 157, 158, 160, 162, 164, 166, 168 or 170;

b) a nucleic acid molecule which comprises at least one polynucleotide of the sequence shown in SEQ ID NO.;

1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 145, 149, 152, 159, 161, 163, 165, 167 or 169;

c) a nucleic acid molecule which encodes a polypeptide according to the sequences SEQ ID NO.;

2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 146, 150, 153, 156, 157, 158, 160, 162, 164, 166, 168 or 170;

d) a nucleic acid molecule with a nucleic acid sequence corresponding to at least one of the sequences according to SEQ ID NO.;

1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 145, 149, 152, 159, 161, 163, 165, 167 or 169 or a nucleic acid molecule derived therefrom by substitution, deletion or insertion which encodes a polypeptide which is at least 40% identical to at least one of the sequences according to SEQ ID NO.;

2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 146, 150, 153, 156, 157, 158, 160, 162, 164, 166, 168 or 170 and is able to bind to keratin;

e) a nucleic acid molecule which encodes a polypeptide which is recognized by a monoclonal antibody directed toward a polypeptide which is encoded by the nucleic acid molecules according to (a) to (c);

f) a nucleic acid molecule encoding for a keratin-binding protein which, under stringent conditions, hybridizes with a nucleic acid molecule according to (a) to (c);

g) a nucleic acid molecule encoding for a keratin-binding protein which can be isolated from a DNA bank using a nucleic acid molecule according to (a) to (c) or part fragments thereof comprising at least 15 nucleotides as probe under stringent hybridization conditions, and

h) a nucleic acid molecule which can be produced by backtranslating one of the amino acid sequences shown in the sequences SEQ ID No.;

NO;

2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 146, 150, 153, 156, 157, 158, 160, 162, 164, 166, 168 or 170.

7. The method according to claim 1, where the linker molecule (iii) has at least two different coupling functionalities.

8. The method according to claim 7, where the linker molecule (iii) has a maleimide group.

9. The method according to claim 8, wherein the linker molecule (iii) is a linker molecule of formula 1

10. The method according to claim 9, where the linker molecule (iii) is a maleimidoalkanol.

11. The method according to claim 10, where the linker molecule (iii) is maleimidopentanol.

12. The method according to claim 1, where a) the keratin-binding polypeptide used comprises one of the sequences according to SEQ ID NO:

2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 146, 150, 153, 156, 157, 158, 160, 162, 166, 168 or 170, and

b) the linker molecule (iii) used is maleimidopentanol, and

c) the effector molecule (i) is 2-(4-diethylamino-2-hydroxybenzoyl)benzoic acid.

14. A keratin-binding effector molecule produced according to claim 12.

19. A dermocosmetic comprising the keratin-binding effector molecule of claim 14.

21. The dermocosmetic of claim 19, wherein the dermocosmetic is a skin protection composition, skincare composition, skin-cleansing composition, hair protection composition, haircare composition, hair-cleansing composition, hair colorant, or a decorative cosmetic.

13. A keratin-binding effector molecule where the effector molecule (i) is coupled indirectly to the keratin-binding polypeptide (ii) via a linker molecule (iii), with the proviso that the linker molecule (iii) is not a maleimide, the keratin-binding polypeptide (ii) does not correspond to the SEQ ID NO.:

166, and effector molecule (ii) is not a fluorescent dye.

18. A dermocosmetic comprising the keratin-binding effector molecule of claim 13.

20. The dermocosmetic of claim 18, wherein the dermocosmetic is a skin protection composition, skincare composition, skin-cleansing composition, hair protection composition, haircare composition, hair-cleansing composition, hair colorant, or a decorative cosmetic.

15. (canceled)

16. (canceled)

17. A compound of formula 2,