Protein hydrolysates

Abstract

The subject of the application is protein hydrolyzates having specific amounts of the amino acids arginine and lysine, which demonstrate improved properties in hair treatment compositions and in cleansing compositions for skin and hair, and also enzymatic methods for the preparation thereof, and the cosmetic compositions comprising same.

Claims

1. A protein hydrolyzate consisting of free and peptide-bound amino acids, wherein 8.0-25 wt % of the amino acids are arginine and 6.0-25 wt % of the amino acids are lysine.

2. The protein hydrolyzate according to claim 1, wherein 5.0 to 25 wt % of the amino acids are lysine.

3. The protein hydrolyzate according to claim 1, wherein 8.2 to 25% of the amino acids are arginine.

4. The protein hydrolyzate according to claim 1, wherein 8.2 to 8.5 wt % of the amino acids are arginine and 6.0 to 6.5 wt % of the amino acids are lysine.

5. A method for preparing a protein hydrolyzate according to claim 1, wherein a mixture of soy and rice proteins in a weight ratio of 2:1 to 4:1 is enzymatically hydrolyzed in the presence of at least one endopeptidase.

6. The method for preparing a protein hydrolyzate according to claim 5, wherein a mixture of soy and rice proteins in a weight ratio of 2:1 to 4:1 is enzymatically hydrolyzed in the presence of at least one endopeptidase and subsequently hydrolyzed once again in the presence of an exopeptidase.

7. A conditioner in a hair treatment composition, comprising the protein hydrolyzate of claim 1.

8. A method for improving the internal hair structure of hair, comprising treating the hair with the protein hydrolyzate of claim 1.

9. A cosmetic composition comprising the protein hydrolyzate according to claim 1, wherein the hydrolyzate is present in an amount of 0.002 to 0.4 wt % based on the active substance content.

10. A cosmetic composition for hair treatment comprising the protein hydrolyzates according to claim 1, wherein the composition additionally comprises cationic or pseudo-cationic surfactants or cationic polymers.

11. A cosmetic composition for cleansing skin and/or hair comprising the protein hydrolyzates according to claim 1, wherein the composition additionally comprises anionic surfactants and amphoteric or zwitterionic surfactants.

Description

EXAMPLES

A) Preparation Examples of the Inventive Protein Hydrolyzates

Inventive Example 1

(1) 5 g of rice protein were suspended in 100 g of water. 15 g of soy protein were added thereto and the pH was adjusted to pH 8.5 using sodium hydroxide solution. 300 mg of protease (from Bacillus Stilistirr; Alcalase® from Novozymes) were subsequently added to the protein mixture and the mixture was stirred for 2 hours.

Inventive Example 2

(2) The mixture obtained according to example 1 was adjusted to pH 7 and subsequently 300 mg of exopeptidase (from Aspergillus oryzae; e.g. Flavourzyme® from Novozymes) were added thereto and the mixture was stirred for a further 4 hours.

(3) The mixtures obtained according to example 1 or 2 were worked up before being used. To this end, they had undissolved protein fragments removed from them using centrifugation or filtration, they were concentrated, and were adjusted to pH 4.2 for stabilization. The active substance content of the protein solution used in water was 20 wt %.

(4) The mixture obtained according to inventive example 2, which had undissolved protein fragments removed therefrom using centrifugation, has the following amino acid composition:

(5) Composition of Amino Acids in Wt %:

(6) TABLE-US-00001 Amino acids Example 2 Alanine 4.48 Arginine 8.44 Aspartic acid 11.83 Cysteine/cystine 1.54 Glutamic acid 19.67 Glycine 4.27 Histidine 2.61 Isoleucine 4.29 Leucine 7.38 Lysine 6.17 Methionine 1.11 Phenylalanine 4.94 Proline 5.03 Serine 5.48 Threonine 3.93 Tyrosine 3.88 Valine 4.95

Comparative Examples

Comparative Example 1: Soy Hydrolyzate (not According to the Invention)

(7) Example 1 was repeated, but only with soy protein and without rice protein.

(8) Amino acid composition: 7.8% arginine, 6.2% lysine

Comparative Example 2: Rice Hydrolyzate (not According to the Invention)

(9) Example 1 was repeated, but only with rice protein and without soy protein.

(10) Amino acid composition: 8.1% arginine, 4.4% lysine

Comparative Example 3: Keratin Hydrolyzate (not According to the Invention)

(11) 100 g of water were initially charged, and adjusted with a base to ˜pH 9.4. Subsequently, 3.75 g of keratin wool were stirred in and, by addition of hydrogen peroxide H.sub.2O.sub.2, the oxidative pretreatment of the wool was started (duration: ˜10 hours). Subsequently, 0.10 g of a protease (e.g. from Bacillus licheniformis; Alcalase from Novozymes), was added. After complete hydrolysis, the remaining water-insoluble wool residues were separated off using filtration and the resulting product was adjusted to pH 4.2 for stabilization.

(12) Amino acid composition: 9.3% arginine, 2.5% lysine

Comparative Example 4: Wheat Hydrolyzate (not According to the Invention)

(13) Example 1 was repeated, but only with wheat protein.

(14) Amino acid composition: 3.1% arginine, 1.4% lysine

B) Use Examples

B1) Hair Repair Using DSC (Differential Scanning Calorimetry) Method

(15) The DSC method is a common thermal analysis method for measuring amounts of absorbed/emitted heat of a sample on heating. Because of the different heat flows between sample and reference during the temperature change program (heating rate), conclusions can be drawn on the denaturation temperatures of the treated hair (sample) relative to untreated hair (reference). The apparatus DSC TA Instruments Q 100 with Autosampler was used. The measurement was carried out according to the method of F. J. Wortmann et al., from: J. Cosmet. Sci., 53, 219-228 (July/August 2002); in a deviation from this, measurement was carried out with a heating rate of 2° C./min.

(16) As sample, Caucasian hair that had been ultra-bleached three times, treated with a 5 wt % aqueous solution of the soy/rice hydrolyzate obtained according to inventive example 2, was tested. 5 wt % aqueous solutions of comparative examples 1 to 4 served as benchmark:

(17) Results:

(18) TABLE-US-00002 DSC in ° C. Inventive 5.68 ± 0.20 Comparative 1 (keratin) 3.73 ± 0.10 Comparative 2 (wheat) 4.06 ± 0.14 Comparative 3 (soy) 3.64 ± 0.10 Comparative 4 (rice) 4.26 ± 0.10

(19) Damaged hair which was treated with the inventive protein hydrolyzate according to example 2 demonstrates a higher decomposition temperature, i.e. an improvement in the denaturation temperature of the hair, which is attributed to an improvement in the hair structure.

B2) Method for Determining Hair Breakage (Hair Breakage Test)

(20) The resistance of the hair to mechanical stress was determined by what is referred to as the “hair breakage test”. To this end, the hair tresses were treated with a conditioner comprising keratin hydrolyzate (composition, see below). After leaving to act for 3 minutes, rinsing was carried out for 1 minute and the process was repeated once. Subsequently, the hair tresses were placed on a grid and dried for 1 hour with a blowdryer. The hair tresses were then combed 50 000 times with a combing machine. The broken hair fragments were weighed and related to the weight of the hair tresses.

(21) The following was tested in a conditioner of the following composition: the values are in wt %:

(22) TABLE-US-00003 Placebo (without Comparative Inventive INCI protein hydrolyzate) example example Lanette ® O Cetyl Stearyl Alcohol 4.5 4.5 4.5 Myritol ® 312 Caprylic/Capric Triglyceride 2 2 2 Dehyquart ® A Cetyl Trimethyl Ammonium 2 2 2 CA Chloride Dehyquart ® F Distearoylethyl Hydroxy- 1.4 1.4 1.4 75 ethylmonium Methosulfate (and) Cetearyl Alcohol Comparative 1 example 1 Example 2 1 Sodium benzoate 0.3 0.3 0.3 Perfume 0.3 0.3 0.3 Water 89.5 88.5 88.5

(23) Statistical significance (p=0.05), calculated according to Tukey's HSD test.

(24) Result:

(25) TABLE-US-00004 Comparative eample 1 Eample 2 Hair breakage vs. placebo No change −20%

(26) The hair treated with the inventive protein hydrolyzate according to example 2 demonstrates 20% less hair breakage than the hair with placebo treatment and than the hair tresses treated with keratin hydrolyzate according to comparative example 1.

B3) Conditioner with Protein Hydrolyzate

(27) A conditioner with the following composition was prepared (values in wt %):

(28) TABLE-US-00005 Conditioner according to INCI the invention Cosmedia ® Triple C Polyquaternium 37(and) Dicaprylylcarbonate 0.40 (and) Lauryl Glucoside Dehyquart ® F 75/ Distearoylethyl Hydroxyethylmonium 2.50 Methosulfate (and) Cetearyl Alcohol Lanette ® O Cetyl stearyl alcohol 3.50 Cutina ® HVG Hydrogenated Vegetable Glycerides 1.00 Cetiol ® C 5 Coco Caprylate 2.00 Cetiol ® SB 45 Butyrospermum Parkii Butter 0.70 Lipofructyl ® Argan LS 9779 Argania Spinosa Kernel Oil 0.20 Water 83.90 Dehyquart ® A-CA Cetyl Trimethyl Ammonium Chloride 3.00 Example 2 2.00 Perfume 0.30 Sodium benzoate 0.50 Citric acid q.s.

B4) Leave-in Serum Conditioner

(29) TABLE-US-00006 INCI Inventive Water 91.53 Example 2 2.00 Melhydran ® LS 9876 Aqua(and)Butylene Glycol(and)Mel 1.00 Extract (and) Glycerin (and) Urea D-Panthenol ® 75 W Panthenol 0.67 Glycerol Glycerol 3.00 Sodium benzoate 0.5 Eumulgin ® CO 40 PEG-40 Hydrogenated Castor Oil 0.5 Copherol ® 1250 C Tocopheryl Acetate 0.5 Perfume 0.3 Citric acid q.s

B5) Application in Hair Shampoos

(30) The following formulations were prepared. The values are in wt %.

(31) TABLE-US-00007 Shampoo 1 Ingredients INCI % Texapon ® N 70 Sodium Laureth Sulfate 14.30 Dehyton ® PK 45 Cocamidopropylbetaine 5.40 Dehyquart ® Guar N Guar Hydroxypropyltrimonium Chloride 0.20 Lamesoft ® Care PEG-4 Distearyl Ether (and) Sodium 3.00 Laureth Sulfate (and) Distearyl Ether (and) Dicaprylyl Ether Dehydol ® LS 2 Laureth-2 1.00 Deo N Water, demin. Aqua 74.60 Example 2 0.50 Sodium Chloride Sodium Chloride 0.50 Sodium Benzoate Sodium Benzoate 0.50 Citric Acid Citric Acid qs

(32) TABLE-US-00008 Shampoo 2 Ingredients INCI % Texapon ® N 70 Sodium Laureth Sulfate 14.30 Dehyton ® PK 45 Cocamidopropylbetaine 5.40 Water, demin. Aqua 77.30 Example 2 0.50 Sodium Chloride Sodium Chloride 2.00 Sodium Benzoate Sodium Benzoate 0.50 Citric Acid Citric Acid qs

(33) TABLE-US-00009 Shampoo 3 Ingredients INCI % Texapon ® N 70 Sodium Laureth Sulfate 14.30 Dehyton ® PK 45 Cocamidopropylbetaine 5.40 Dehyquart ® Guar N Guar Hydroxypropyltrimonium Chloride 0.20 Water, demin. Aqua 76.70 Example 2 1.00 Sodium Chloride Sodium Chloride 1.60 Perfume Cotton Touch Perfume 0.30 Sodium Benzoate Sodium Benzoate 0.50 Citric Acid Citric Acid qs

(34) Comb tests on dry hair demonstrated a reduction in the required combability of 20% compared to the placebo shampoo without inventive protein hydrolyzate.

(35) TABLE-US-00010 Shampoo4 “Extra Care” Ingredients INCI % Water Water 68.12 Dehyquart ® Guar N Guar Hydroxypropyltrimonium 0.20 Chloride Dehyton ® PK 45 Cocamidopropylbetaine 2.70 Luviquat ® UltraCare Polyquaternium-44 1.15 Texapon ® N 70 Sodium Laureth Sulfate 12.90 Plantapon ® LC 7 Laureth-7 Citrate 1.00 Plantacare ® 818 UP Coco-Glucoside 0.40 Lamesoft ® PO 65 Coco-Glucoside, Glyceryl Oleate 5.00 Cetiol ® LDO Dicaprylyl Ether, Lauryl Alcohol 0.70 Example 2 2.00 D-Panthenol 75 W Panthenol 1.33 Euperlan ® PK 710 Benz Glycol Distearate, Sodium Laureth 3.00 Sulfate, Cocamide MEA Sodium Benzoate Sodium Benzoate 0.50 Perfume Perfume qs Magnesium Chloride Magnesium Chloride 1.00 Hexahydrate Citric Acid (50% solution) Citric Acid qs Sodium Chloride Sodium Chloride qs

(36) TABLE-US-00011 Shampoo 5 “Thickening Shampoo” Ingredients INCI % Texapon ® N 70 Sodium Laureth 14.30 Sulfate Dehyton ® PK 45 Cocamidopropylbetaine 5.40 Plantapon ® LC 7 Laureth-7 Citrate 2.00 Gluadin ® WQ PP Laurdimonium Hydroxypropyl 0.50 Hydrolyzed Wheat Protein Example 2 2.00 D-Panthenol 75 W Panthenol 1.33 Euperlan ® PK 710 Benz Glycol Distearate, Sodium Laureth 3.00 Sulfate, Cocamide MEA Perfume Perfume qs Water Water 68.97 Sodium Benzoate Sodium Benzoate 0.50 Magnesium Chloride Magnesium Chloride 1.00 Hexahydrate Dehydol LS 2 Deo N Laureth-2 1.00 Citric Acid (50% solution) Citric Acid qs Sodium Chloride Sodium Chloride qs

B6) Conditioner as Gel

(37) TABLE-US-00012 Conditioning Gel Ingredients INCI % Water, demin. Aqua 80.90 Example 2 1.50 Ethanol Alcohol 15.00 Dehyquart ® Guar HP Guar Hydroxypropyltrimonium 1.50 Chloride Citric Acid Citric Acid 0.30 Luviquat ® Supreme AT 1 Polyquaternium-68 2.00 Perfume Cotton Touch Perfume 0.30

B7) Styling Mousse

(38) TABLE-US-00013 Ingredients INCI % Water, demin. Aqua 82.70 Example 2 2.00 Luviquat ® Supreme AT 1 Polyquaternium-68 7.50 Luviquat ® PQ 11 AT 1 Polyquaternium-11 7.50 Cremophor ® A 25 Ceteareth-25 0.50 Perfume Cotton Touch Perfume 0.10 Sodium Benzoate Sodium Benzoate 0.30 Citric Acid (20% solution) Citric Acid 1.40

B8) Conditioner for Natural Cosmetics

(39) TABLE-US-00014 Ingredients INCI % Plantaquat ® NC Cetearyl Alcohol (and) Lecithin (and) 9.0 Sodium Cetearyl Sulfate (and) Olus Oil [EU] Lanette ® O Cetearyl Alcohol 2.0 Cetiol ® C5 Coco-Caprylate 2.0 Cegesoft ® SB Butyspermum Parkii Butter 2.0 Example 2 2.0 Sodium Benzoate Sodium Benzoate 0.5 Perfume Perfume q.s. Water Water add 100

B9) Hair and Body Wash

(40) TABLE-US-00015 Hair and Body Wash -Cleansing composition 1 for natural cosmetics Ingredients INCI % Water, demin. Water 68.20 Sulfopon ® 1216 G Sodium Coco-Sulfate 8.00 Plantacare ® 818 UP Coco-Glucoside 13.30 Euperlan ® GREEN Lauryl Glucoside (and) Stearyl Citrate 2.00 Lamesoft ® PO 65 Coco-Glucoside (and) Glyceryl Oleate 2.00 Example 2 0.50 Sodium Benzoate Sodium Benzoate 0.50 Perfume Perfume q.s. Apple juice 100%, direct Pyrus Malus (Apple) Juice 3.00 juice Citric Acid (50%) Citric Acid 1.20 Sodium Chloride Sodium Chloride 1.30

(41) TABLE-US-00016 Hair and Body Wash -Cleansing composition 2 for natural cosmetics Ingredients INCI % Water, demin. Water 70.10 Sulfopon ® 1216 G Sodium Coco-Sulfate 8.00 Plantacare ® 818 UP Coco-Glucoside 13.30 Lamesoft ® PO 65 Coco-Glucoside (and) Glyceryl Oleate 2.00 Example 2 0.50 Sodium Benzoate Sodium Benzoate 0.50 Perfume Perfume q.s. Apple juice 100%, direct Pyrus Malus (Apple) Juice 3.00 juice Citric Acid (50%) Citric Acid 1.20 Sodium Chloride Sodium Chloride 1.30