Hybrid polymers containing polysaccharide polymers and peptide chains

Abstract

The present invention relates to hybrid polymers comprising polysaccharide polymers and peptide chains, blends, hydrogels and preparations comprising said hybrid polymers, and the use of said hybrid polymers as rheological modifiers.

Description

Technical Field

[0001] The present invention relates to hybrid polymers containing polysaccharide polymers and peptide chains, blends containing such hybrid polymers, hydrogels and preparations, and the use of said hybrid polymers as rheology modifiers.

Background Art

[0002] The cleaning and care of the skin and hair are very important for general hygiene, for example for the removal of unwanted substances such as sebum, oil, dirt, makeup, etc., or for moisturizing, complexion or protection. Many cosmetics require a specific minimum viscosity in order to achieve ease of application to the substrate and / or retention on the substrate to be treated. Many cosmetics contain viscosity increasing agents or rheology modifiers. These are often referred to as thickening agents, thickeners or gelling agents. Examples of thickening agents used in cosmetics or personal care products include polyethylene glycol, polyacrylic acid, vegetable gums, polycarboxylate (Carbopol), and homopolymers and copolymers based on 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS).

[0003] Many of the ingredients used in cosmetics are traditionally derived from crude oil. Environmental and economic factors limit the use of products derived from this limited resource. It is desirable to identify more sustainable and biodegradable but gentle and effective materials. In fact, consumers are interested in natural products, including products with a high proportion of ingredients derived from natural components and / or renewable materials. Consumers perceive products derived from natural materials as being gentler and more environmentally friendly.

[0004] Ingredients derived from renewable materials have various other advantages, such as increased biodegradability and more sustainable availability. Compounds derived from plant-based resources are particularly useful because the compound source can be simply regrown.

[0005] WO2018 / 108663 (Patent Document 1), WO2018 / 108664 (Patent Document 2), WO2018 / 108665 (Patent Document 3), and WO2018 / 108667 (Patent Document 4) disclose water-soluble and / or water-swellable hybrid polymers comprising polysaccharide polymers and synthetic polymers. [Prior art documents] [Patent Documents]

[0006] [Patent Document 1] WO2018 / 108663 [Patent Document 2] WO2018 / 108664 [Patent Document 3] WO2018 / 108665 [Patent Document 4] WO2018 / 108667 [Overview of the Initiative] [Problems that the invention aims to solve]

[0007] There is a continuing need for polymer-based rheological modifiers that can provide the superior performance of modern polymers, offering enhanced biodegradability and greater sustainable use of natural-based polymers. [Means for solving the problem]

[0008] This study revealed that hybrid polymers containing polysaccharide polymers and peptide chains are useful as rheological modifiers.

[0009] This invention relates to a hybrid polymer comprising the following: (a) Polysaccharide polymers; and (b) Peptide chain.

[0010] The hybrid polymers of the present invention exhibit excellent performance as rheological modifiers. They form hydrogels with desired properties. Advantageously, the hybrid polymers of the present invention have a high renewable carbon content and are readily biodegradable; that is, they are sustainable.

[0011] The hybrid polymer of the present invention comprises (a) a polysaccharide polymer and (b) a peptide chain. Preferably, the total amount of (a) the polysaccharide polymer and (b) the peptide chain in the hybrid polymer is at least 50% by weight, more preferably at least 60% by weight, more preferably at least 70% by weight, more preferably at least 75% by weight, more preferably at least 80% by weight, more preferably at least 85% by weight, even more preferably at least 90% by weight, even more preferably at least 95% by weight, even more preferably at least 97% by weight, even more preferably at least 98% by weight, and particularly preferably at least 99% by weight, based on the total weight of the hybrid polymer.

[0012] The hybrid polymer of the present invention includes a polysaccharide polymer.

[0013] Preferably, the polysaccharide polymer is water-soluble and / or water-swellable. In at least one embodiment, when immersed in water, the polysaccharide polymer absorbs water and / or forms a gel or gum. In at least one embodiment, the polysaccharide polymer is a natural gum. Natural gums are useful because they are generally water-soluble due to the presence of an excess number of OH groups that form hydrogen bonds with water molecules. In at least one embodiment, the polysaccharide polymer is a natural gum derived from plants.

[0014] In preferred embodiments, the polysaccharide polymer is selected from dextran, dextrin, hyaluronic acid, chitosan, xanthan gum, fenugreek gum, tara gum, locust bean gum, carrageenan, guar gum, alginate, agar, agarose, starch, amylose, amylopectin, tragacanth gum, tamarind kernel gum, arabic gum, cherry gum, karaya gum, okra gum, cassia gum, chicle gum, konjac gum, ghati gum, pectin, sclerotium gum, gellan gum, paramylon, paramylum, curdlan, cellulose, diutan gum, inulin, derivatives thereof, and mixtures thereof.

[0015] In a more preferred embodiment, the polysaccharide polymer is selected from dextran, dextrin, xanthan gum, tara gum, locust bean gum, carrageenan, guar gum, alginate, starch, amylose, amylopectin, tamarind kernel gum, gum arabic, karaya gum, konjac gum, pectin, sclerotium gum, gellan gum, diutan gum, inulin, derivatives thereof, and mixtures thereof.

[0016] In a more preferred embodiment, the polysaccharide polymer is selected from dextran, dextrin, xanthan gum, tara gum, locust bean gum, carrageenan, guar gum, starch, amylose, amylopectin, konjac gum, inulin, derivatives thereof, and mixtures thereof.

[0017] In a particularly preferred embodiment, the polysaccharide polymer is dextran.

[0018] Derivatives of the above polysaccharide polymers can also be used. "Derivative" means that the polysaccharide polymer has been subjected to one or more suitable physical, chemical or enzymatic processes by which the polysaccharide polymer is converted into the derivative of the polysaccharide polymer. Examples of such processes include the following: - Acid treatment of the polysaccharide polymer by reaction with an acid (e.g., hydrochloric acid, phosphoric acid or sulfuric acid) - Alkali treatment of the polysaccharide polymer by reaction with a base (e.g., sodium hydroxide or potassium hydroxide) - Bleached polysaccharide polymers by reaction with peracetic acid, hydrogen peroxide, sodium hypochlorite, sulfur dioxide, sulfite, potassium permanganate or ammonium persulfate - Enzymatic modification of the polysaccharide polymer by treatment with an enzyme - Oxidized polysaccharide polymers by oxidation (e.g., by sodium hypochlorite) - Acetylated polysaccharide polymers by esterification with, for example, an anhydride - Hydroxypropyl polysaccharide polymers by reaction with propylene oxide - Hydroxyethyl polysaccharide polymers by reaction with ethylene oxide - Carboxymethylation of the polysaccharide polymer - Glycol polysaccharide polymers Preferred derivatives of the polysaccharide polymer are selected from carboxymethyl polysaccharides, hydroxyethyl polysaccharides, carboxymethyl hydroxyethyl polysaccharides, carboxypropyl polysaccharides, glycol polysaccharides, and mixtures thereof.

[0019] Examples of preferred derivatives of the polysaccharide polymer are carboxymethyl cellulose, hydroxyethyl cellulose, carboxymethyl hydroxyethyl cellulose, carboxypropyl cellulose, carboxymethyl chitosan, glycol chitosan or mixtures thereof.

[0020] In a preferred embodiment, the polysaccharide polymer has a weight-average molecular weight of 3,000 to 1,200,000,000 g / mol, preferably 10,000 to 5,000,000 g / mol, more preferably 30,000 to 2,000,000 g / mol, even more preferably 50,000 to 1,000,000 g / mol, and particularly preferably 100,000 to 500,000 g / mol.

[0021] In a preferred embodiment, the polysaccharide polymer has a weight-average molecular weight of 3,000 to 1,000,000 g / mol, preferably 10,000 to 500,000 g / mol, more preferably 20,000 to 300,000 g / mol, even more preferably 30,000 to 200,000 g / mol, and particularly preferably 40,000 to 150,000 g / mol.

[0022] In a preferred embodiment, the polysaccharide polymer has a weight-average molecular weight of 15,000 to 75,000 g / mol, preferably 20,000 to 70,000 g / mol, more preferably 20,000 to 60,000 g / mol, more preferably 30,000 to 60,000 g / mol, even more preferably 30,000 to 50,000 g / mol, and even more preferably 35,000 to 45,000 g / mol, for example, 40,000 g / mol.

[0023] In a preferred embodiment, the polysaccharide polymer has a weight-average molecular weight of 100,000 to 200,000 g / mol, preferably 120,000 to 180,000 g / mol, more preferably 130,000 to 170,000 g / mol, and even more preferably 140,000 to 160,000 g / mol, for example, 150,000 g / mol.

[0024] In a preferred embodiment, the polysaccharide polymer has a weight-average molecular weight of at least 180,000 g / mol, preferably at least 200,000 g / mol.

[0025] When used herein, the molecular weight, including the weight-average molecular weight of the polysaccharide polymer, is determined using gel permeation chromatography (GPC). The PSS SECcurity consists of a pump, autosampler, and column oven. 2 GPC experiments are performed using the apparatus. A SUPREMA LIN XL column with dimensions of 300 × 8 mm and an average particle size of 10 μm is used at a flow rate of 1.0 mL / min and a column temperature of 25°C. 0.1 M NaNO3 is used as the eluent.

[0026] The hybrid polymer of the present invention contains peptide chains.

[0027] The peptide is preferably a dipeptide, tripeptide, oligopeptide, polypeptide, or protein. What peptides and proteins are is known to those skilled in the art. A dipeptide contains two amino acids. A tripeptide contains three amino acids. The oligopeptide preferably contains 4 to 9 amino acids. The polypeptide preferably contains at least 10 amino acids, more preferably 10 to 99 amino acids. A protein preferably contains at least 100 amino acids, more preferably 100 to 30,000 amino acids, even more preferably 100 to 1,000 amino acids, and particularly preferably 100 to 300 amino acids. The peptide may also be a hydrolyzed protein. What hydrolyzed proteins, also called protein hydrolysates, are is known to those skilled in the art. Hydrolyzed proteins can be obtained by hydrolyzing a protein into peptides and amino acids. Such hydrolysis can be chemical or enzymatic. Enzymatic hydrolysis is preferred because it allows for controlled hydrolysis by targeting specific peptide bonds.

[0028] The amino acids in a peptide chain can have any stereochemical orientation. For example, each amino acid can be independently an L-amino acid or a D-amino acid. In one embodiment, all amino acids in the peptide chain are L-amino acids. In another embodiment, all amino acids in the peptide chain are D-amino acids.

[0029] The amino acids in the peptide chain may optionally be in salt form. Those skilled in the art know that this can depend, for example, on the environment of the peptide chain. For example, an acidic environment may result in the protonation of the basic functional group of the amino acid side chain. For example, a basic environment may result in the deprotonation of the acidic functional group of the amino acid side chain.

[0030] In a preferred embodiment, the peptide chains form aggregates with each other.

[0031] In a preferred embodiment, the peptide chains are self-assembling. While we do not wish to be constrained by theory, the peptide chains may form non-covalent bonds with each other. The peptide chains may form supramolecular assemblies, which may result in non-covalent bonds or supramolecular crosslinks that can lead to gelation. In one embodiment, the peptide chains may form nanofiber or nanofibril structures. In one embodiment, the peptide chains may form beta-sheets.

[0032] In a preferred embodiment, at least 50%, preferably at least 60%, more preferably at least 70%, even more preferably at least 80%, even more preferably at least 90%, even more preferably at least 95%, and particularly preferably at least 98% of the amino acids in the peptide chain are protein-constituting amino acids. In a preferred embodiment, all amino acids in the peptide chain are protein-constituting amino acids. Preferably, the protein-constituting amino acids are selected from leucine, isoleucine, phenylalanine, valine, tyrosine, tryptophan, lysine, aspartic acid, glutamic acid, arginine, histidine, methionine, serine, threonine, glycine, alanine, proline, cysteine, asparagine, glutamine, selenocysteine, and pyrrolicin. More preferably, the protein constituent amino acids are selected from leucine, isoleucine, phenylalanine, valine, tyrosine, tryptophan, lysine, aspartic acid, glutamic acid, arginine, histidine, methionine, serine, threonine, glycine, alanine, proline, cysteine, asparagine, and glutamine.

[0033] The peptide chain may optionally contain additional amino acids F. In several embodiments, up to 50%, preferably up to 40%, more preferably up to 30%, even more preferably up to 20%, even more preferably up to 10%, even more preferably up to 5%, and particularly preferably up to 2% of the amino acids in the peptide chain are additional amino acids F. In several embodiments, the peptide chain does not contain additional amino acids F. Preferably, the additional amino acids F are amino acids other than protein constituent amino acids. Preferably, the additional amino acids F are fluorine-containing amino acids, hydroxyl-containing amino acids other than protein constituent amino acids, boronic acid-containing amino acids, anthracenyl-containing amino acids, and C2-C2 amino acids other than protein constituent amino acids. 10 amino acids having alkyl groups, C2-C 10 Amino acids having an alkenyl group, and C2-C 10The further amino acid F is selected from amino acids having an alkynyl group. Preferably, the further amino acid F is selected from azidohomoalanine, acridinylalanine, phenylselenocysteine, sulfoserine, p-iodophenylalanine, bipyridylalanine, dansylalanine, o-nitrobenzylcysteine, 7-nitroindolinyl amino acids, propargylglycine, azidonorleucine, 5-bromotryptophan, L-4'-deoxy-4'-iodophenylalanine, tetrazinalanine, dipyridyltetraginserine, hydroxyproline, betaalanine, citrulline, ornithine, norleucine, 3-nitrotyrosine, nitroarginine, and pyroglutamic acid.

[0034] In a preferred embodiment, at least 25%, preferably at least 30%, of the amino acids in the peptide chain are identical. In a preferred embodiment, at least 25%, preferably at least 30%, of the amino acids in the peptide chain are identical and preferably selected from hydrophobic amino acids. In a more preferred embodiment, at least 25%, preferably at least 30%, of the amino acids in the peptide chain are identical and preferably selected from hydrophobic amino acids. Preferably, the hydrophobic amino acid is selected from leucine, isoleucine, phenylalanine, valine, tyrosine, and tryptophan. More preferably, the hydrophobic amino acid is selected from leucine, isoleucine, phenylalanine, and valine. Even more preferably, the hydrophobic amino acid is selected from leucine, isoleucine, and phenylalanine. Particularly preferably, the hydrophobic amino acid is isoleucine.

[0035] In a preferred embodiment, at least 15%, preferably at least 20%, of the amino acids in the peptide chain are identical. In a preferred embodiment, at least 15%, preferably at least 20%, of the amino acids in the peptide chain are identical and preferably selected from charged amino acids. In a more preferred embodiment, at least 15%, preferably at least 20%, of the amino acids in the peptide chain are identical and preferably selected from charged amino acids. Preferably, the charged amino acid is selected from lysine, aspartic acid, glutamic acid, arginine, and histidine. More preferably, the charged amino acid is selected from lysine, aspartic acid, and glutamic acid. Particularly preferably, the charged amino acid is lysine.

[0036] In a preferred embodiment, at least 10%, preferably at least 20%, of the amino acids in the peptide chain are selected from amino acids having an amide group in their side chain. Preferably, the amino acids having an amide group in their side chain are selected from asparagine, glutamine, and mixtures thereof.

[0037] In a preferred embodiment, the peptide chain contains at least one methionine.

[0038] In a preferred embodiment, the peptide chain contains at least one amino acid selected from serine, threonine, and mixtures thereof. In a more preferred embodiment, the peptide chain contains at least one serine.

[0039] In a preferred embodiment, at least 40%, preferably at least 45%, of the amino acids in the peptide chain are selected from hydrophobic amino acids. In a preferred embodiment, at least 40%, preferably at least 45%, of the amino acids in the peptide chain are selected from hydrophilic amino acids. In a more preferred embodiment, at least 40%, preferably at least 45%, of the amino acids in the peptide chain are selected from hydrophobic amino acids, and at least 40%, preferably at least 45%, of the amino acids in the peptide chain are selected from hydrophilic amino acids.

[0040] Preferably, the hydrophobic amino acid is selected from leucine, isoleucine, phenylalanine, valine, tyrosine, tryptophan, glycine, alanine, proline, and methionine. More preferably, the hydrophobic amino acid is selected from leucine, isoleucine, phenylalanine, valine, tyrosine, tryptophan, and methionine. Even more preferably, the hydrophobic amino acid is selected from leucine, isoleucine, phenylalanine, valine, and methionine. Particularly preferably, the hydrophobic amino acid is selected from isoleucine, phenylalanine, and methionine.

[0041] Preferably, the hydrophilic amino acid is selected from serine, threonine, cysteine, asparagine, glutamine, lysine, aspartic acid, glutamic acid, arginine, and histidine. More preferably, the hydrophilic amino acid is selected from serine, threonine, cysteine, asparagine, glutamine, lysine, aspartic acid, and glutamic acid. Particularly preferably, the hydrophilic amino acid is selected from serine, cysteine, asparagine, glutamine, lysine, and glutamic acid.

[0042] In a preferred embodiment, at least 80%, preferably at least 90%, of the amino acids in the peptide chain are arranged so that hydrophobic amino acids and hydrophilic amino acids alternate.

[0043] In a particularly preferred embodiment, the peptide chain is selected from the peptide chain of formula P1, the peptide chain of formula P2, the peptide chain of formula P3, their salts, their tautomers, salts of their tautomers, and mixtures thereof:

[0044] [ka] Peptide P1 may be characterized by the above structure, or it may be a salt thereof, a tautomer thereof, or a salt of a tautomer thereof.

[0045] Therefore, peptide P1 has the structure CKIKISQINM (Sequence ID 1) in single-letter notation.

[0046] The amino acids of peptide P1 can have any stereochemical orientation. For example, each amino acid can be independently an L-amino acid or a D-amino acid. In one embodiment, all amino acids of peptide P1 are L-amino acids. In one embodiment, all amino acids of peptide P1 are D-amino acids.

[0047] Peptide P2 may be characterized by the above structure, or it may be a salt thereof, a tautomer thereof, or a salt of a tautomer thereof.

[0048] Therefore, peptide P2 has the structure CEIEISQINM (Sequence ID 2) in single-letter notation.

[0049] The amino acids of peptide P2 can have any stereochemical orientation. For example, each amino acid can be independently an L-amino acid or a D-amino acid. In one embodiment, all amino acids of peptide P2 are L-amino acids. In one embodiment, all amino acids of peptide P2 are D-amino acids.

[0050] Peptide P3 may be characterized by the above structure, or it may be a salt thereof, a tautomer thereof, or a salt of a tautomer thereof.

[0051] Peptide P3 has the structure CKFKFQF (Sequence ID 3) in single-letter notation.

[0052] The amino acids of peptide P3 can have any stereochemical orientation. For example, each amino acid can be independently an L-amino acid or a D-amino acid. In one embodiment, all amino acids of peptide P3 are L-amino acids. In one embodiment, all amino acids of peptide P3 are D-amino acids.

[0053] In a preferred embodiment, the peptide chain is a depsipeptide chain. A depsipeptide is known to those skilled in the art. A depsipeptide is a peptide in which one or more amide groups in the peptide chain are replaced by ester groups. Preferably, one to three amide groups in the peptide chain are replaced by ester groups. More preferably, one or two amide groups in the peptide chain are replaced by ester groups. Particularly preferably, one amide group in the peptide chain is replaced by an ester group. In a preferred embodiment, the ester group is part of the structural unit of formula D:

[0054] [ka] In a preferred embodiment, the peptide chain contains at least one structural unit of formula D. Preferably, the structural unit of formula D is located in the center of the peptide chain.

[0055] In a particularly preferred embodiment, the peptide chain is selected from the peptide chain of formula P4, the peptide chain of formula P5, their salts, their tautomers, salts of their tautomers, and mixtures thereof:

[0056] [ka] Depsipeptide P4 may be characterized by the above structure, or it may be a salt thereof, a tautomer thereof, or a salt of a tautomer thereof. Therefore, depsipeptide P4 may also be represented by the following structure: [CKIKI]-O-CH2-C(NH2)-CO-[QINM] Therefore, depsipeptide P4 contains two peptide portions, each represented by a single letter: sequence CKIKI (sequence number 4) and QINM (sequence number 5). In the structure above, the peptide portions are indicated by parentheses ("[...]").

[0057] Depsipeptide P5 may be characterized by the above structure, or it may be a salt thereof, a tautomer thereof, or a salt of a tautomer thereof. Therefore, depsipeptide P5 may also be represented by the following structure: [CEIEI]-O-CH2-C(NH2)-CO-[QINM] Therefore, the depsipeptide P5 contains two peptide moieties, each represented by a single letter: sequence CIEII (sequence number 6) and QINM (sequence number 5). In the structure above, the peptide moieties are indicated by parentheses ("[...]").

[0058] The amino acids in the peptide portions of depsipeptides P4 and P5 can have any stereochemical orientation. For example, each amino acid can be independently an L-amino acid or a D-amino acid. In one embodiment, all amino acids in one peptide portion are L-amino acids. In one embodiment, all amino acids in one peptide portion are D-amino acids. In one embodiment, all amino acids in both peptide portions of a single depsipeptide are D-amino acids. In one embodiment, all amino acids in both peptide portions of a single depsipeptide are L-amino acids.

[0059] The structural units of formula D can be rearranged into the structural units of formula S:

[0060] [ka] A peptide chain containing structural units of formula D can be rearranged into a peptide chain containing structural units of formula S. Such rearrangement can be initiated by adjusting the pH, for example, by increasing the pH. In one embodiment, rearrangement can be initiated by adjusting the pH from an acidic pH (e.g., pH 2) to a higher pH, for example, a neutral pH (e.g., pH 7). Rearrangement can change the geometric shape of the peptide chain, for example, from a twisted to a linear shape. Rearrangement can change the solubility of the peptide chain. Rearrangement can cause aggregation of the peptide chain.

[0061] In one embodiment, the peptide chain contains at least one structural unit of formula L:

[0062] [ka] Preferably, the structural unit of formula L is located in the center of the peptide chain.

[0063] The structural units of formula L can be rearranged into the structural units of formula M:

[0064] [ka] A peptide chain containing structural units of formula L can be rearranged into a peptide chain containing structural units of formula M. Such rearrangement can be initiated by adjusting the pH, for example, by increasing the pH. In one embodiment, rearrangement can be initiated by adjusting the pH from an acidic pH (e.g., pH 2) to a higher pH, for example, a neutral pH (e.g., pH 7). Rearrangement can change the geometric shape of the peptide chain, for example, from a twisted to a linear shape. Rearrangement can change the solubility of the peptide chain. Rearrangement can cause aggregation of the peptide chain.

[0065] In one embodiment, one or more amide groups in the peptide chain are replaced by thioester groups. Preferably, one to three amide groups in the peptide chain are replaced by thioester groups. More preferably, one or two amide groups in the peptide chain are replaced by thioester groups. Particularly preferably, one of the amide groups in the peptide chain is replaced by a thioester group. In a preferred embodiment, the thioester group is part of the structural unit of formula T:

[0066] [ka] In one embodiment, the peptide chain contains at least one structural unit of formula T. Preferably, the structural unit of formula T is located in the center of the peptide chain.

[0067] The structural units of formula T can be rearranged into the structural units of formula C:

[0068] [ka] A peptide chain containing structural units of formula T can be rearranged into a peptide chain containing structural units of formula C. Such rearrangement can be initiated by adjusting the pH, for example, by increasing the pH. In one embodiment, rearrangement can be initiated by adjusting the pH from an acidic pH (e.g., pH 2) to a higher pH, for example, a neutral pH (e.g., pH 7). Rearrangement can change the geometric shape of the peptide chain, for example, from a twisted to a linear shape. Rearrangement can change the solubility of the peptide chain. Rearrangement can cause aggregation of the peptide chain.

[0069] The peptide chain may optionally contain N-protected versions of structural units of formulas D, L, or T. Cleavage of the N-protecting group can induce rearrangement to structural units of formulas S, M, or C, respectively. The N-protecting group can be cleaved, for example, by UV irradiation. Therefore, rearrangement can be induced, for example, by UV radiation.

[0070] The peptide chain may optionally be interrupted by ether or polyether crosslinking, by ethylene glycol or polyethylene glycol crosslinking, or by C3-C6 alkylene crosslinking.

[0071] The peptide chain may be of plant, animal, or human origin, or it may be, for example, a viral peptide, protein or fragment thereof, or a bacterial peptide, protein or fragment thereof. In one preferred embodiment, the peptide chain is of plant origin.

[0072] In a preferred embodiment, the peptide chain is selected from pea protein, hydrolyzed pea protein, soy protein, hydrolyzed soy protein, broad bean protein, hydrolyzed broad bean protein, chickpea protein, for example aquafaba, hydrolyzed chickpea protein, wheat protein, hydrolyzed wheat protein, rice protein, hydrolyzed rice protein, whey protein, hydrolyzed whey protein, baobab protein, hydrolyzed baobab protein, collagen protein, hydrolyzed collagen protein, plant collagen-like protein, hydrolyzed plant collagen-like protein, hemp seed protein, hydrolyzed hemp seed protein, jojoba protein, hydrolyzed jojoba protein, keratin protein, hydrolyzed keratin protein, lupine protein, hydrolyzed lupine protein, oat protein, hydrolyzed oat protein, quinoa protein, hydrolyzed quinoa protein, and mixtures thereof.

[0073] In a more preferred embodiment, the peptide chain is selected from pea protein, hydrolyzed pea protein, soy protein, hydrolyzed soy protein, broad bean protein, hydrolyzed broad bean protein, chickpea protein, for example aquafaba, hydrolyzed chickpea protein, wheat protein, hydrolyzed wheat protein, rice protein, hydrolyzed rice protein, whey protein, hydrolyzed whey protein, oat protein, hydrolyzed oat protein, quinoa protein, hydrolyzed quinoa protein, and mixtures thereof.

[0074] In a more preferred embodiment, the peptide chain is selected from pea protein, hydrolyzed pea protein, oat protein, hydrolyzed oat protein, and mixtures thereof.

[0075] In one embodiment, the peptide chain is selected from proteins. Preferably, the peptide chain is selected from pea protein, soybean protein, broad bean protein, chickpea protein, for example, aquafaba, wheat protein, rice protein, whey protein, baobab protein, collagen protein, plant collagen-like protein, hemp seed protein, jojoba protein, keratin protein, lupine protein, oat protein, quinoa protein, and mixtures thereof. More preferably, the peptide chain is selected from pea protein, soybean protein, broad bean protein, chickpea protein, for example, aquafaba, wheat protein, rice protein, whey protein, oat protein, quinoa protein, and mixtures thereof. Even more preferably, the peptide chain is selected from pea protein, oat protein, and mixtures thereof.

[0076] In another embodiment, the peptide chain is selected from hydrolyzed proteins. Preferably, the peptide chain is selected from hydrolyzed pea protein, hydrolyzed soy protein, hydrolyzed broad bean protein, hydrolyzed chickpea protein, hydrolyzed wheat protein, hydrolyzed rice protein, hydrolyzed whey protein, hydrolyzed baobab protein, hydrolyzed collagen protein, hydrolyzed plant collagen-like protein, hydrolyzed hemp seed protein, hydrolyzed jojoba protein, hydrolyzed keratin protein, hydrolyzed lupine protein, hydrolyzed oat protein, hydrolyzed quinoa protein, and mixtures thereof. More preferably, the peptide chain is selected from hydrolyzed pea protein, hydrolyzed soy protein, hydrolyzed broad bean protein, hydrolyzed chickpea protein, hydrolyzed wheat protein, hydrolyzed rice protein, hydrolyzed whey protein, hydrolyzed oat protein, hydrolyzed quinoa protein, and mixtures thereof. Even more preferably, the peptide chain is selected from hydrolyzed pea protein, hydrolyzed oat protein, and mixtures thereof.

[0077] In a preferred embodiment, the peptide chain contains 2 to 5000, preferably 3 to 4000, more preferably 4 to 3000, more preferably 5 to 2000, more preferably 5 to 1000, even more preferably 6 to 500, even more preferably 6 to 100, and particularly preferably 7 to 50, for example, 7 to 10 amino acids.

[0078] In a preferred embodiment, the peptide chain contains 2 to 100, preferably 3 to 50, more preferably 4 to 30, even more preferably 5 to 20, and particularly preferably 6 to 15, for example, 7 to 10 amino acids.

[0079] In one embodiment, the peptide chain contains 8 or fewer amino acids. In another embodiment, the peptide chain contains 10 or more amino acids.

[0080] In a preferred embodiment, the peptide chain contains 100 to 5000, preferably 200 to 4000, more preferably 300 to 3000, even more preferably 400 to 2000, and particularly preferably 500 to 1000 amino acids.

[0081] In a preferred embodiment, the peptide chain has a weight-average molecular weight of 150 to 600,000 g / mol, preferably 200 to 400,000 g / mol, more preferably 300 to 300,000 g / mol, more preferably 400 to 200,000 g / mol, more preferably 500 to 100,000 g / mol, even more preferably 600 to 50,000 g / mol, even more preferably 700 to 10,000 g / mol, and particularly preferably 750 to 5,000 g / mol, for example 800 to 1,500 g / mol.

[0082] In a preferred embodiment, the peptide chain has a weight-average molecular weight of 150 to 10000 g / mol, preferably 300 to 5000 g / mol, more preferably 500 to 4000 g / mol, even more preferably 600 to 3000 g / mol, and particularly preferably 700 to 2000 g / mol, for example, 800 to 1500 g / mol.

[0083] In a preferred embodiment, the peptide chain has a weight-average molecular weight of 10,000 to 600,000 g / mol, preferably 20,000 to 400,000 g / mol, more preferably 30,000 to 300,000 g / mol, even more preferably 40,000 to 200,000 g / mol, and particularly preferably 50,000 to 100,000 g / mol.

[0084] In a preferred embodiment, the peptide chain has a molecular weight of 150 to 600,000 g / mol, preferably 200 to 400,000 g / mol, more preferably 300 to 300,000 g / mol, more preferably 400 to 200,000 g / mol, more preferably 500 to 100,000 g / mol, even more preferably 600 to 50,000 g / mol, even more preferably 700 to 10,000 g / mol, and particularly preferably 750 to 5,000 g / mol, for example 800 to 1,500 g / mol.

[0085] In a preferred embodiment, the peptide chain has a molecular weight of 150 to 10000 g / mol, preferably 300 to 5000 g / mol, more preferably 500 to 4000 g / mol, even more preferably 600 to 3000 g / mol, and particularly preferably 700 to 2000 g / mol, for example, 800 to 1500 g / mol.

[0086] In a preferred embodiment, the peptide chain has a molecular weight of 10,000 to 600,000 g / mol, preferably 20,000 to 400,000 g / mol, more preferably 30,000 to 300,000 g / mol, even more preferably 40,000 to 200,000 g / mol, and particularly preferably 50,000 to 100,000 g / mol.

[0087] The peptides used in this invention may be commercially available, described in the literature, manufactured biotechnically, or produced according to methods known in the art. For example, the peptides used in this invention can be manufactured by solid-phase peptide synthesis or liquid-phase peptide synthesis. Generally, peptides are manufactured from corresponding amino acids using known techniques (e.g., carboxylic acid activation, coupling reagents) and protecting group strategies. For example, the peptides used in this invention, such as P1-P5, can be manufactured according to the method described in Gacanin, J. et al., Adv. Mater. 2019, 31, 1805044 (Autonomous Ultrafast Self-Healing Hydrogels by pH-Responsive Functional Nanofiber Gelators as Cell Matrices).

[0088] In one preferred embodiment, the peptide chain is of synthetic origin. In one preferred embodiment, the peptide chain is of natural origin.

[0089] In a preferred embodiment, the peptide chain is linked to the polysaccharide polymer via a linker. Any linker can be used.

[0090] In a preferred embodiment, the linker arises from the incorporation of a compound having at least two electrophilic functional groups.

[0091] Preferably, the compound having at least two electrophilic functional groups has 1 to 20 carbon atoms and 0 to 10 heteroatoms, more preferably 2 to 12 carbon atoms and 1 to 7 heteroatoms, even more preferably 3 to 9 carbon atoms and 2 to 5 heteroatoms, and particularly preferably 4 to 7 carbon atoms and 3 to 4 heteroatoms.

[0092] Preferably, at least two electrophilic functional groups are selected from epoxy groups, C=C double bonds, carbonyl groups, C=C double bonds substituted with at least one carbonyl group, and mixtures thereof.

[0093] Preferably, the compound having at least two electrophilic functional groups is selected from glycidyl methacrylate, maleimide, and mixtures thereof. Particularly preferred is glycidyl methacrylate as the compound having at least two electrophilic functional groups.

[0094] In one embodiment, the linker is not obtained from the incorporation of glycidyl methacrylate.

[0095] In one embodiment, the hybrid polymer of the present invention comprises (a) a polysaccharide polymer, (b) a peptide chain, and (c) optionally a linker. Preferably, the total amount of (a) the polysaccharide polymer, (b) the peptide chain, and (c) optionally a linker in the hybrid polymer is at least 50% by weight, more preferably at least 60% by weight, more preferably at least 70% by weight, more preferably at least 75% by weight, more preferably at least 80% by weight, more preferably at least 85% by weight, even more preferably at least 90% by weight, even more preferably at least 95% by weight, even more preferably at least 97% by weight, even more preferably at least 98% by weight, and particularly preferably at least 99% by weight, based on the total weight of the hybrid polymer.

[0096] In one embodiment, the hybrid polymer of the present invention comprises (a) a polysaccharide polymer, (b) a peptide chain, and (c) a linker. Preferably, the total amount of (a) the polysaccharide polymer, (b) the peptide chain, and (c) the linker in the hybrid polymer is at least 50% by weight, more preferably at least 60% by weight, more preferably at least 70% by weight, more preferably at least 75% by weight, more preferably at least 80% by weight, more preferably at least 85% by weight, even more preferably at least 90% by weight, even more preferably at least 95% by weight, even more preferably at least 97% by weight, even more preferably at least 98% by weight, and particularly preferably at least 99% by weight, based on the total weight of the hybrid polymer.

[0097] In a preferred embodiment, 1 to 100%, preferably 3 to 50%, more preferably 5 to 40%, even more preferably 8 to 30%, and particularly preferably 10 to 25% of the monosaccharide units of the polysaccharide polymer are optionally modified with peptide chains via linkers. Modification of the monosaccharide units / polysaccharide polymer typically occurs via modifiable groups of the monosaccharide units / polysaccharide polymer. Preferably, the modifiable group is a hydroxyl group. The modifiable group may also be, for example, an amino group. In the case of dextran, the monosaccharide units are glucose units, and the modifiable group is a hydroxyl group.

[0098] In a preferred embodiment, the peptide chain is optionally linked to the polysaccharide polymer via a linker, either via the N-terminal amino acid or the C-terminal amino acid of the peptide chain.

[0099] Any amino acid can be used as the N-terminal or C-terminal amino acid of the peptide chain. In one embodiment, the N-terminal or C-terminal amino acid of the peptide chain is cysteine. In one embodiment, the N-terminal amino acid of the peptide chain is cysteine. In one embodiment, the C-terminal amino acid of the peptide chain is cysteine.

[0100] In one embodiment, the N-terminal amino acid of the peptide chain is not lysine.

[0101] In one embodiment, the C-terminal amino acid of the peptide chain is not cysteine. In one embodiment, the C-terminal amino acid of the peptide chain is selected from hydrophobic amino acids. Preferably, the hydrophobic amino acids are selected from leucine, isoleucine, phenylalanine, valine, tyrosine, tryptophan, glycine, alanine, proline, and methionine. In one embodiment, the C-terminal amino acids of the peptide chain are selected from methionine and phenylalanine.

[0102] The N-terminal amino acid of the peptide chain may be optionally capped, for example, it may be optionally acetylated. The N-terminal amino acid of the peptide chain may be optionally protected with a protecting group, for example, a fluorenyl methoxycarbonyl (Fmoc) group. The N-terminal amino acid of the peptide chain may be optionally capped (for example, with a C3-C6 alkyl group). The C-terminal amino acid of the peptide chain may be optionally capped, for example, it may be optionally amidated.

[0103] This invention also relates to blends including the following: (a) 1 to 99% by weight, preferably 20 to 80% by weight, more preferably 30 to 70% by weight, even more preferably 40 to 65% by weight, and the hybrid polymer of the present invention, based on the total weight of the blend; and (b) One or more polysaccharide polymers and / or one or more peptide chains in an amount of 1 to 99% by weight, preferably 20 to 80% by weight, more preferably 30 to 70% by weight, even more preferably 35 to 60% by weight, and especially preferably 40 to 50% by weight, based on the total weight of the blend.

[0104] Preferred hybrid polymers are further described above.

[0105] In a preferred embodiment, the blend of the present invention includes: (a) 1 to 99% by weight, preferably 20 to 80% by weight, more preferably 30 to 70% by weight, even more preferably 40 to 65% by weight, and the hybrid polymer of the present invention, based on the total weight of the blend; and (b) One or more polysaccharide polymers in an amount of 1 to 99% by weight, preferably 20 to 80% by weight, more preferably 30 to 70% by weight, even more preferably 35 to 60% by weight, and especially preferably 40 to 50% by weight, based on the total weight of the blend.

[0106] Preferred polysaccharide polymers are further described above.

[0107] Preferably, the polysaccharide polymer is selected from dextran, dextrin, hyaluronic acid, chitosan, xanthan gum, fenugreek gum, tara gum, locust bean gum, carrageenan, guar gum, alginate, agar, agarose, starch, amylose, amylopectin, tragacanth gum, tamarind kernel gum, gum arabic, cherry gum, karaya gum, okra gum, cassia gum, chicle gum, konjac gum, ghati gum, pectin, sclerotium gum, gellan gum, paramylon, paramylum, curdlan, cellulose, diutan gum, inulin, derivatives thereof, and mixtures thereof.

[0108] More preferably, the polysaccharide polymer is selected from dextran, dextrin, xanthan gum, tara gum, locust bean gum, carrageenan, guar gum, alginate, starch, amylose, amylopectin, tamarind kernel gum, gum arabic, karaya gum, konjac gum, pectin, sclerotium gum, gellan gum, diutan gum, inulin, derivatives thereof, and mixtures thereof.

[0109] More preferably, the polysaccharide polymer is selected from dextran, dextrin, xanthan gum, tara gum, locust bean gum, carrageenan, guar gum, starch, amylose, amylopectin, konjac gum, inulin, derivatives thereof, and mixtures thereof.

[0110] Particularly preferred is the polysaccharide polymer being xanthan gum.

[0111] In a preferred embodiment, the blend of the present invention includes: (a) 1 to 99% by weight, preferably 20 to 80% by weight, more preferably 30 to 70% by weight, even more preferably 40 to 65% by weight, and the hybrid polymer of the present invention, based on the total weight of the blend; and (b) One or more peptides in an amount of 1 to 99% by weight, preferably 20 to 80% by weight, more preferably 30 to 70% by weight, even more preferably 35 to 60% by weight, and especially preferably 40 to 50% by weight, based on the total weight of the blend.

[0112] Preferred peptides are further described above.

[0113] The present invention also relates to the use of the hybrid polymer or blend of the present invention as a rheological modifier. In preferred embodiments, the rheological modifier is a thickener.

[0114] Preferably, the hybrid polymer or blend of the present invention is used as a rheological modifier in cosmetic preparations. In preferred embodiments, the cosmetic preparation is a skincare preparation or a haircare preparation. More preferably, the hybrid polymer or blend of the present invention is used as a rheological modifier in skincare preparations or haircare preparations. Particularly preferably, the hybrid polymer or blend of the present invention is used as a rheological modifier in skincare preparations. Also particularly preferably, the hybrid polymer or blend of the present invention is used as a rheological modifier in haircare preparations.

[0115] Preferably, the hybrid polymer or blend of the present invention is used as a thickening agent in cosmetic preparations. In preferred embodiments, the cosmetic preparation is a skincare preparation or a haircare preparation. More preferably, the hybrid polymer or blend of the present invention is used as a thickening agent in skincare preparations or haircare preparations. Particularly preferably, the hybrid polymer or blend of the present invention is used as a thickening agent in skincare preparations. Also particularly preferably, the hybrid polymer or blend of the present invention is used as a thickening agent in haircare preparations.

[0116] The present invention also relates to a hydrogel comprising the hybrid polymer or blend of the present invention, water, optionally a pH adjuster, and optionally an ionic strength adjuster.

[0117] In one embodiment, the hydrogel comprises the hybrid polymer or blend of the present invention, water, and a pH adjuster or ionic strength adjuster.

[0118] In one embodiment, the hydrogel comprises the hybrid polymer or blend of the present invention, water, and a pH adjuster.

[0119] In one embodiment, the hydrogel comprises the hybrid polymer or blend of the present invention, water, and an ionic strength modifier.

[0120] In one embodiment, the hydrogel comprises the hybrid polymer or blend of the present invention, water, a pH adjuster, and an ionic strength adjuster.

[0121] Preferred hybrid polymers are further described above.

[0122] In a preferred embodiment, the hydrogel contains 0.1 to 10% by weight, preferably 0.2 to 5% by weight, more preferably 0.3 to 3% by weight, even more preferably 0.4 to 2% by weight, and particularly preferably 0.5 to 1% by weight of a hybrid polymer or blend, based on the total weight of the hydrogel.

[0123] Preferably, the hydrogel contains a pH adjuster. The pH adjuster may be, for example, an acid, a base, a buffer, or a combination thereof. Preferred acid examples include hydrochloric acid, acetic acid, trifluoroacetic acid, citric acid, formic acid, vitamin C, or a combination thereof. Preferred base examples include sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate (sodium hydrogen carbonate), potassium bicarbonate (potassium hydrogen carbonate), or a combination thereof. Preferred buffer examples include phosphate buffer, citrate buffer, acetate buffer, or a combination thereof.

[0124] A particularly preferred pH adjusting agent is a buffer. A particularly preferred buffer is a phosphate buffer. A particularly preferred pH adjusting agent is a phosphate buffer.

[0125] Optionally, the hydrogel may contain an ionic strength modifier. The ionic strength modifier may be, for example, a salt. Preferred salts include sodium chloride, potassium chloride, or a combination thereof. A particularly preferred ionic strength modifier is sodium chloride.

[0126] Preferably, the hydrogel has a pH of 3 to 9, more preferably 4 to 8.5, even more preferably 5 to 8, and particularly preferably 5.5 to 7.5.

[0127] In a preferred embodiment, the hydrogel further comprises an active ingredient. Preferably, the active ingredient is selected from vitamins, humectants, anti-aging agents, anti-wrinkle agents, anti-inflammatory agents, amino acids, peptides, and mixtures thereof. More preferably, the active ingredient is selected from vitamins, humectants, anti-aging agents, anti-wrinkle agents, anti-inflammatory agents, and mixtures thereof.

[0128] Furthermore, the present invention relates to a preparation comprising the following: (a) 0.1 to 10% by weight, preferably 0.2 to 5% by weight, more preferably 0.3 to 3% by weight, even more preferably 0.4 to 2% by weight, and particularly preferably 0.5 to 1% by weight of the hybrid polymer or blend of the present invention, based on the total weight of the preparation; and (b) One or more further components in an amount of 90-99.9% by weight, preferably 95-99.8% by weight, more preferably 97-99.7% by weight, even more preferably 98-99.6% by weight, and particularly preferably 99-99.5% by weight, based on the total weight of the preparation.

[0129] In a preferred embodiment, the preparation is a cosmetic preparation, preferably a skincare preparation or a haircare preparation. In one preferred embodiment, the preparation is a skincare preparation. In another preferred embodiment, the preparation is a haircare preparation.

[0130] In a preferred embodiment, the preparations include shampoo, body wash, facial cleanser, face mask, bubble bath, intimate wash, bath oil, cleansing milk, micellar water, makeup remover, cleansing wipes, hair mask, fragrance, liquid soap, shaving soap, shaving foam, cleansing foam, day cream, anti-aging cream, body milk, body lotion, body mousse, face serum, eye cream, sunscreen lotion, sun cream, face cream, aftershave lotion, pre-shaving cream, hair removal cream, skin whitening gel, self-tanning cream, anti-acne gel, mascara, foundation, primer, concealer, blush, bronzer, blemish cream, and more. BB cream, eyeliner, night cream, eyebrow gel, highlighter, lip stain, hand sanitizer, hair oil, nail polish remover, skin conditioner, hair conditioner, hair styling gel, hair styling cream, anti-frizz serum, scalp treatment, hair colorant, split end solution, deodorant, antiperspirant, baby cream, insect repellent, hand cream, sunscreen gel, foot cream, exfoliator, body scrub, cellulite treatment, bar soap, cuticle cream, lip balm, hair treatment, eyeshadow, bath additive, body mist, eau de toilette, mouthwash, toothpaste, lubricating gel, moisturizer, serum, toner, aqua sherbet The group consists of sorbet, cream gel, styling mousse, dry shampoo, lipstick, lip gloss, body oil, shower milk, illuminator, lip crayon, hairspray, combing cream, and sunblock.

[0131] In a more preferred embodiment, the preparations include body wash, facial cleanser, face mask, bubble bath, intimate wash, bath oil, cleansing milk, micellar water, makeup remover, cleansing wipes, fragrance, liquid soap, shaving soap, shaving foam, cleansing foam, day cream, anti-aging cream, body milk, body lotion, body mousse, face serum, eye cream, sunscreen lotion, sun cream, face cream, aftershave lotion, pre-shaving cream, hair removal cream, skin whitening gel, self-tanning cream, anti-acne gel, mascara, foundation, primer, concealer, blush, bronzer, blush The group consists of BB cream, eyeliner, night cream, eyebrow gel, highlighter, lip stain, hand sanitizer, nail polish remover, skin conditioner, scalp treatment, deodorant, antiperspirant, baby cream, insect repellent, hand cream, sunscreen gel, foot cream, exfoliator, body scrub, cellulite treatment, bar soap, cuticle cream, lip balm, eyeshadow, bath additive, body mist, eau de toilette, mouthwash, toothpaste, lubricating gel, moisturizer, serum, toner, aqua sherbet, cream gel, lipstick, lip gloss, body oil, shower milk, illuminator, lip crayon, and sunblock.

[0132] In a more preferred embodiment, the preparation may include body wash, facial cleanser, face mask, intimate wash, cleansing milk, micellar water, makeup remover, cleansing wipes, liquid soap, shaving soap, shaving foam, cleansing foam, day cream, anti-aging cream, body milk, body lotion, body mousse, face serum, eye cream, sunscreen lotion, sun cream, face cream, aftershave lotion, pre-shaving cream, hair removal cream, skin whitening gel, self-tanning cream, mascara, and face cream. The group consists of foundations, primers, concealers, blushes, bronzers, blemish balm (BB) creams, night creams, highlighters, lip stains, hand sanitizers, nail polish removers, skin conditioners, deodorants, antiperspirants, baby creams, insect repellents, hand creams, sunscreen gels, foot creams, exfoliators, body scrubs, cellulite treatments, cuticle creams, lip balms, lubricating gels, moisturizers, toners, cream gels, lipsticks, lip glosses, body oils, shower milks, illuminators, lip crayons, and sunblocks.

[0133] The following examples are intended to illustrate the present invention and are not intended to limit it thereto. [Examples]

[0134] example P1-P5 refer to peptides (P1-P3) and depsipeptides (P4-P5). When the pH was increased to neutral, P1 was formed from depsipeptide P4. When the pH was increased to neutral, P2 was formed from depsipeptide P5. Furthermore, P1 and P2 can be synthesized directly, for example, via solid-phase peptide synthesis, without the intermediate depsipeptide step. All amino acids used are L-amino acids. P1-P5 were prepared according to methods known in the art.

[0135] Unless otherwise specified, RT refers to room temperature (20-25°C).

[0136] 1 Synthesis 1.1 Dextran-methacrylate (1a, 1b, 2): Functionalization of dextran with glycidyl methacrylate (GMA, 3) (4, 5) In a typical reaction, dextran (15 g, molecular weight (MW) 40,000 g / mol, 4) was placed in a flask under a nitrogen atmosphere and dissolved by adding dry dimethyl sulfoxide (DMSO, 90 mL), stirring until dissolution was complete (approximately 30 minutes). The mixture was heated to 40°C in an oil bath with stirring to aid dissolution. 4-(dimethylamino)pyridine (DMAP, 2.25 g, 0.2 equivalents, 6) was separately dissolved in dry DMSO (24 mL). Subsequently, GMA (4.65 mL, 0.4 equivalents, 3) was added to the DMAP (6) solution, and the mixture was added to the dextran solution. The reaction mixture was stirred at 40°C for 20 hours. All steps were carried out under a nitrogen atmosphere. The reaction mixture was purified by dialysis (molecular weight cutoff (MWCO) 3.5 kDa) in deionized water for 5 days, which included water changes every 2-3 hours on the first day and twice a day for the remainder of the day. After freeze-drying for 4 days, product (1a) was obtained as a white solid. 1 GMA functionalization was quantified by 1H NMR (DMSO-d6). This reaction modified approximately 21% of the glucose units in the dextran with methacrylate functional groups. The above method was applied to dextran with various molecular weights, e.g., MW 40 kg / mol (4) and 150 kg / mol (5), yielding their respective methacrylated species, i.e., 1a, 1b, or 2. Variations in dextran molecular weight and degree of methacrylate were achieved by changing the free form, as summarized in Table 1.

[0137] [Table 1]

[0138] 1.2 Peptide-dextran hybrids General synthetic explanation - Using a hybrid H1 based on peptide P4 (CKIKI*SQINM, where the asterisk (*) indicates the position of the ester bond within the depsipeptide: C: cysteine, K: lysine, I: isoleucine, S: serine, Q: glutamine, N: asparagine, M: methionine) as an example: GMA-functionalized dextran (1, 40 kDa, 10 mg) was dissolved in 0.1% trifluoroacetic acid (7, TFA) in water (Milli-Q, 0.2 mL) (A). Separately, tris-(2-carboxyethyl)-phosphine (8, TCEP, 2.4 mg, 0.15 equivalents) was dissolved in 0.1% TFA / H2O (0.3 mL) and added to peptide (P4, 21.0 mg, 0.3 equivalents). The mixture (B) was incubated in a shaker at room temperature for 30 minutes. Solution B was then added to solution A and stirred overnight (18 h) at 66°C under a nitrogen atmosphere (shaker 1000 rpm). The reaction product was purified by ultrafiltration in 0.1% TFA / H2O at 4000 rpm and 25°C using a Vivaspin tube (5x, MWCO 5 kDa, PES). After 3 days of freeze-drying, the hybrid (H1) was obtained as a white powder. For larger batch sizes, the freeze-drying time was extended. The amounts of biopolymer, peptide, TCEP, and solvent used were varied to achieve different functionalization efficiencies, as shown in Table 2.

[0139] [Table 2]

[0140] 1.3 Hydrogel Manufacturing a) Peptide-dextran hybrid i) Direct addition of buffer to hybrid (solid): The hydrogel was prepared by directly adding the respective amounts of buffer to the hybrid powder. For example, to obtain a 1 wt% hydrogel, 0.3 mg of 30 μL of 100 mM phosphate buffer pH 7.4 was added to the hybrid. Gelation occurred upon gentle mixing.

[0141] ii) pH switch of pre-dissolved hybrid (acidic solution): Preferably, the hydrogel was obtained by pre-dissolving the hybrid in an acidic solution and adjusting the pH to approximately pH 7. For example, to obtain a 1 wt% hydrogel, 0.3 mg of the hybrid was dissolved in 15 μL of 0.1% TFA / H2O (pH 2). Subsequently, 15 μL of 300 mM phosphate buffer pH 9 was added to adjust the pH to neutral (pH 7), and gelation began. No further mixing or stirring was necessary. Addition in the reverse order was also possible. Alternatively, 0.8 mg of the hybrid was dissolved in 20 μL of 0.1% TFA / H2O (pH 2), and gelation began when 60 μL of 300 mM phosphate buffer pH 9 was added to adjust the pH to neutral.

[0142] iii) Optimized pH switch for pre-dissolved hybrid (acidic solution): Most preferably, the hydrogel was obtained by pre-dissolving the hybrid in an acidic solution as described in ii) and subsequently incubating it at approximately 55°C (50-60°C) for approximately 30-45 minutes. Subsequently, the pH was adjusted to approximately pH 7 by adding a buffer preheated to the same temperature as the hybrid, i.e., 55°C. For example, to obtain a 1 wt% hydrogel, 20 mg of the hybrid was dissolved in 1 mL of 0.1% TFA / H2O (pH 2) and heated at 55°C for 30-45 minutes. Subsequently, gelation began when 1 mL of 300 mM phosphate buffer pH 9 (55°C) was added to adjust the pH to neutral. No further mixing or stirring was required.

[0143] iv) Variations of iii) for incorporating preservatives: Optimized pH switch for pre-dissolved hybrid (acidic solution) with gel preservation: Hydrogels were obtained by pre-dissolving the hybrid in an acidic solution, as described in iii). Subsequently, the pH was adjusted to approximately pH 7 by adding a buffer preheated to the same temperature as the hybrid, i.e., 55°C. The buffer was supplemented with a preservative (e.g., Nipaguard DMDMH Plus, 0.6 wt%). For example, to obtain a 1 wt% hydrogel, 30 mg of the hybrid was dissolved in 1.5 mL of 0.1% TFA / H2O (pH 2) and heated at 55°C for 30-45 minutes. Subsequently, 1.5 mL of 300 mM phosphate buffer pH 9, supplemented with Nipaguard DMDMH Plus (0.6 wt%, 7.8 μL) (55°C), was added to adjust the pH to neutral, and gelation began. No further mixing or stirring was required.

[0144] v) Alternative buffers can be used to adjust the pH to neutral. For example, instead of 300 mM phosphate buffer (pH 9), 300 mM phosphate buffer pH 7 can be used to switch the pH from acidic to neutral. Similarly, other acids, such as vitamin C, can be used to dissolve the hybrid at pH 2.

[0145] b) Peptide-dextran hybrid and polysaccharide complex hydrogel i. Xanthan gum: Optimized pH switch Hydrogels were obtained by pre-dissolving the hybrid in an acidic solution and then incubating at 55°C (50-60°C) for 30-45 minutes (see 1.3aiii). To prepare the composite hydrogel, xanthan gum (X.gum) hydrogel was added. Subsequently, the pH was adjusted to approximately pH 7 by adding phosphate buffer. For example, to obtain a 0.8 wt% composite hydrogel (see Table 3:1), 10 mg of the hybrid was dissolved in 1 mL of 0.1% TFA / H2O (pH 2) and heated at 55°C for 30-45 minutes. Next, 1 mL of 1 wt% X.gum hydrogel was added to the solution and mixed thoroughly. Subsequently, 0.5 mL of 300 mM phosphate buffer pH 9 was added to adjust the pH to neutral, and gelation of the hybrid began. Further mixing or stirring was not necessary.

[0146] The most preferred hybrid to X.gum ratio was 1:1 (by weight). However, alternative ratios may be selected to adjust the gel properties, as summarized in Table 3:

[0147] [Table 3]

[0148] ii. Dextran (1) Dextran (4, e.g., 10 mg or 20 mg) was dissolved together with the hybrid (20 mg) in 0.1% TFA / H2O (total volume 1 mL, pH 2), heated to 55°C, and 300 mM phosphate buffer pH 9 (1 mL) was added to adjust the pH to neutral. The procedure was carried out according to 1.3aiii.

[0149] (2) The hybrid (e.g., 10 mg or 20 mg) was dissolved in 0.1% TFA / H2O (1 mL, pH 2), heated to 55°C, and dextran (20 mg) pre-dissolved in 300 mM phosphate buffer pH 9 (1 mL) was added to adjust the pH to neutral. The procedure was carried out according to 1.3aiii.

[0150] (3) Solution A was prepared by adding 0.1% TFA / H2O (7.5 μL) to H1 (0.15 mg). Solution B was prepared by adding 0.1% TFA / H2O (7.5 μL) to dextran (4, 0.15 mg). Next, Solution B was added to Solution A while mixing. Subsequently, phosphate buffer (15 μL, 300 mM, pH 9) was added while mixing. The procedure was carried out according to 1.3aii.

[0151] iii. Hyaluronic acid Hydrogels were obtained by pre-dissolving the hybrid in an acidic solution and then incubating at 55°C (50-60°C) for 30-45 minutes (see 1.3aiii). Hyaluronic acid was added to prepare the composite hydrogel. Subsequently, the pH was adjusted to approximately pH 7 by adding phosphate buffer. For example, to obtain the composite hydrogel, 10 mg of hybrid H6 was dissolved in 1 mL of 0.1% TFA / H2O (pH 2) and heated at 55°C for 30-45 minutes. Next, 1 mL of pre-dissolved hyaluronic acid (10 mg, H2O) was added to the solution and mixed thoroughly. Subsequently, 0.5 mL of 300 mM phosphate buffer pH 9 was added to adjust the pH to neutral, and gelation of the hybrid began. Further mixing or stirring was not necessary.

[0152] 2 Method / Characteristics analysis 2.1 NMR To quantify the functionalization rate of polysaccharides in GMA or peptides, 1 1H NMR was performed. DMSO-d6 was used for the analysis. Spectra were recorded using a Bruker 300MHz or 400MHz NMR spectrometer.

[0153] 2.2 Rheology The rheological properties of the hydrogels and their preparations were analyzed using a DHR3 rheometer (TA Instruments) equipped with a temperature controller. Experiments were performed using: a) an 8 mm parallel plate geometry with a solvent reservoir to prevent hydrogel drying, where a 30 μL volume of hydrogel resulted in a gap size of approximately 0.50 mm; b) a 20 mm cone plate geometry (2.007°) with a solvent reservoir to prevent hydrogel drying; or c) a 40 mm cone plate geometry. In general, the mechanical properties were analyzed at 20°C. Furthermore, the mechanical properties at temperatures of 5–60°C were also analyzed.

[0154] We used several different measurement settings: (i) Time sweep: It was found that the linear viscoelastic region includes the range of 0.05–1% strain and 1–1.6 Hz frequency. Therefore, vibration time sweep measurements were performed at fixed strain and fixed frequency within this range, at different temperatures and time intervals.

[0155] (ii) Strain sweep 1: A vibrational strain sweep was performed at a fixed frequency of 1 Hz at 25°C or as indicated, for strains between 0.01 and 1000%.

[0156] (iii) Strain sweep 2: A vibrational strain sweep was performed at 20°C with a fixed frequency of 1 Hz for strains between 0.1% and 100%.

[0157] (iv) Thixotropy Measurement 1: To analyze the self-healing ability of the hydrogel, a series of combined measurements were performed: a vibration-strain sweep at 25°C with a fixed frequency of 1 Hz (0.01–1000%) followed by a vibration-type sweep measurement at 25°C for 800 or 1000 seconds with a fixed strain of 0.1% and a frequency of 1 Hz. In particular, the repair efficiency of the hydrogel and preparations was quantified from the storage modulus obtained from this experiment. In this regard, the mean hydrogel recovery during the initial hydrogel recovery (first data point obtained, approximately 13 seconds) and the subsequent vibration-time sweep (0.1% strain, 800 seconds) was analyzed for each strain sweep.

[0158] (v) Thixotropy Measurement 2: To analyze the self-healing ability of the hybrid X.gum composite hydrogel, a series of measurements were performed, consisting of a vibrational strain sweep (0.01-100%) at 20°C with a fixed frequency of 1 Hz, followed by a vibrational type sweep measurement of 0.1% fixed strain and a frequency of 1 Hz for 300 seconds. In particular, the repair efficiency of the hydrogel and preparation was quantified from the storage modulus obtained from this experiment. In connection with this, the average hydrogel recovery during the subsequent vibrational time sweep (0.1% strain, 800 seconds) was analyzed for each strain sweep.

[0159] (vi) Rheology recovery test: A combination of measurements using continuous vibration time sweeps alternating between high strain (200%, 120 seconds) and low strain (0.1%, 120 seconds) at a fixed frequency of 1 Hz and 20°C. Repeated up to 10 times.

[0160] (vii) Gradient change in flow rate: Rheological experiment with continuous flow rate. Gradient change in flow rate from initial stress 0 to 60 Pa over 180 seconds at 20°C or 25°C.

[0161] (viii) Temperature sweep: Vibrational temperature sweep measurements were performed with fixed strain (0.05%) and fixed frequency (1 Hz). First, a temperature sweep was performed with a starting temperature of 5°C and an ending temperature of 60°C, followed by a reverse temperature sweep from 60°C to 5°C; both measurements were performed in 5°C temperature steps.

[0162] (ix) Temperature analysis: Continuous measurement of vibration time sweep (0.05% strain, 1 Hz, 2400 seconds, 20°C); temperature sweep from 20°C to 60°C (start temperature 20°C and end temperature 60°C with a temperature step of 5°C, 0.05% strain, 1 Hz), reverse temperature sweep from 60°C to 20°C (start temperature 60°C and end temperature 20°C with a temperature step of 5°C, 0.05% strain, 1 Hz), time sweep (immersion time 180 seconds, 0.05% strain, 1 Hz, 400 seconds, 20°C), thixotropy measurement (continuous alternating vibration amplitude sweep 0.01%~1000%, 1 Hz, time sweep 0.05% strain, 1 Hz, 1000 seconds; both at 20°C).

[0163] Furthermore, we conducted several experiments using these experimental setups: x) Study on the effects of peptide graft amino acid sequences on gel mechanical properties such as stiffness, crossover strain, thixotropy, and recovery (variations of hydrophobic amino acids including positively or negatively charged amino acids, or aromatic residues). xi) Study of the effect of peptide graft density on gel mechanical properties such as stiffness, crossover strain, thixotropy, and recovery (H1-H8, e.g., 0%-20% glucose units per dextran molecule with peptide functionalization) xii) Study of the effect of polysaccharide backbone length on gel mechanical properties such as stiffness, crossover strain, thixotropy, and recovery (e.g., dextran 40 kg / mol or 150 kg / mol) xiii) Study on the effect of temperature on gel mechanical properties such as stiffness, crossover strain, thixotropy, and recovery (T5℃~60℃) xiv) Study on the effect of pH on gel mechanical properties such as stiffness, crossover strain, thixotropy, and recovery (pH 5 to pH 9) To evaluate the effect of pH changes on the mechanical properties of hydrogels, hydrogels were prepared at pH 7 via a pH switch as described above. Next, the hydrogels were washed with buffer until the desired pH was reached, and rheological properties were analyzed. For example, a 1 wt% hydrogel was prepared by introducing 0.3 mg of H1 into 15 μL of 0.1% TFA / H2O and inducing gelation by adding 15 μL of 300 mM phosphate buffer pH 9. Gelation was completed. After 10 minutes, the gel was washed with 150 mM phosphate buffer pH 7 (for control), 100 mM acetate buffer pH 4.75, or 150 mM phosphate buffer pH 9.

[0164] xv) Study of the effect of salt on gel mechanical properties such as stiffness, crossover strain, thixotropy, and recovery (NaCl, 0 or 5% by weight) Furthermore, in addition to hydrogels derived from peptide-dextran hybrids, some of the above experiments were also performed on composite materials: xvi) Study of the effects of adding peptides (ungrafted) with different amino acid sequences on gel mechanical properties such as stiffness, crossover strain, thixotropy, and recovery. Here, peptides were introduced either as a newly prepared peptide solution (monomer) in the pH 2 phase or as pre-incubated fibrils in the pH 9 phase.

[0165] xvii) Study on the effects of adding free polysaccharides on gel mechanical properties such as stiffness, crossover strain, thixotropy, and recovery. 2.3 Tilting / Overturn Test in Glass Vials A 2.5 mL volume of hydrogel was prepared in a 5 mL total volume small glass vial according to the preparation protocol described herein. During gelation, the glass vial was tilted 180° to demonstrate the presence or absence of flow in the liquid or weak gel / preparation.

[0166] 2.4 Uptake of active substances a) Vitamin C (10, final concentration 5% by weight) i) Most preferably, H6 (10 mg) and vitamin C (125 mg) were dissolved in H2O (Milli-Q, 0.5 mL), and xanthan gum (9, 1 wt% in water, 1 mL) was added while mixing. Subsequently, phosphate buffer (300 mM, pH 9, 2 mL) was added to the mixture to adjust the pH to neutral.

[0167] ii) H6 (10 mg) was dissolved in vitamin C solution (125 mg in 0.1% TFA / H2O, 1 mL). Xanthan gum (1% by weight in water, 1 mL) was added while mixing. Subsequently, phosphate buffer (300 mM, pH 9, 2 mL) was added to the mixture to adjust the pH to neutral.

[0168] iii) H6 (10 mg) was dissolved in vitamin C solution (125 mg in 0.1% TFA / H2O, 1 mL) and heated at approximately 55°C for 30 minutes. Xanthan gum (1% by weight in water, 1 mL) was added while mixing. Subsequently, phosphate buffer (300 mM, pH 9, 2 mL) was added to the mixture to adjust the pH to neutral.

[0169] b) Peptides i. Monomers Solution A was prepared by adding 0.1% TFA / H2O (7.5 μL) to H1 (0.15 mg). Solution B was prepared by adding 0.1% TFA / H2O (7.5 μL) to either P4 or P5 (0.15 mg), respectively. Next, Solution B was added to Solution A while mixing. Subsequently, phosphate buffer (15 μL, 300 mM, pH 9) was added while mixing. During the pH switch process, the depsipeptides rearrange into their linear counterparts; that is, P1 is formed from P4, while P2 is formed from P5 by the switch, and aggregation begins.

[0170] ii. Pre-assembled fibrils Solution A was prepared by adding 0.1% TFA / H2O (7.5 μL) to H1 (0.15 mg). Solution B was prepared by adding 0.1% TFA / H2O (7.5 μL) to P4 (0.15 mg). Solution B was then introduced into phosphate buffer (15 μL, 300 mM, pH 9) while mixing, and incubated at room temperature for 5 minutes without stirring or shaking. During the pH switch process, the depsipeptides rearrange into their linear counterparts; that is, the switch causes P1 to form from P4, and aggregation begins. Next, this mixture was added to solution A while mixing.

[0171] 2.5 Cell viability The cell viability of hydrogels and their components was evaluated by performing the CellTiter-Glo® Luminescent Cell Viability Assay. The test was performed using hydrogel extracts pre-incubated in cell medium, referring to DIN EN ISO 10993-5 (Biologische Beurteilung von Medizinprodukten - Teil 5: Pruefungen auf In-vitro-Zytotoxizitaet). Extracts were prepared and cells were seeded on day 1:1 wt% hydrogel (H1 or H5 / H6) was prepared according to 1.3ai or 1.3aii and left for gelation (15 minutes, room temperature). Cell medium (DMEM with 1% penicillin / streptomycin, 1% non-essential amino acids (NEAA), and 5% fetal bovine serum (FCS) fully added; 200 μL) was added. Soluble components (dextran 40 kg / mol(4), dextran-GMA(1), P4) were pre-dissolved in buffer according to 1.3ai or 1.3aii, similar to the gel hybrid, to obtain 1% or 0.5% by weight, and cell medium was added (200 μL). For the control, cell medium was used as a blind value (Blindwert), the positive control was doxorubicin 500 μM (in cell medium), the control was buffer (same concentration as in the extract preparation: 20 μL buffer (100 mM phosphate buffer pH 7.4) + 200 μL cell medium), and cell medium was used as a blank (no cells). The extract, sample solution and control were incubated overnight at 37°C. Furthermore, cells were seeded according to the manufacturer's protocol (half-area plate, 5000 A549 cells per well (50 μL; 1 × 10⁶)). -5Cells (per mL), fully added DMEM, incubated overnight (37°C, 5% CO2). On day 0, the cell medium was removed from the wells and replaced with extracts, samples or controls in solution (50 μL per well), which were then added to the cells (triple-duplicate) and incubated for 24 hours (37°C, 5% CO2). On day 1, the CellTiter-Glo® Luminescent Cell Viability Assay was performed according to the manufacturer's protocol.

[0172] 2.6 Hydrogel Degradation Assay: Biodegradation A biodegradation test (Biological Oxygen Demand (BODn) method) was conducted according to OECD 301F. During the measurement, the BOD-Direct Plus System was placed in an incubator heated to 20°C. Sodium benzoate (50 mg / L) was used as the standard. The concentration of the test substance (H5) was approximately 50 mg / L. The test was conducted for 28 days using inoculum in a sealed bottle.

[0173] 2.7 Long-term temperature stability of hydrogels (macroscopic) Hydrogel (H6, 1 mL, 1 wt%) was prepared in a glass vial according to 1.3aiv and incubated at 4°C, room temperature (approximately 20°C), or 37°C.

[0174] 2.8 Molecular weight The molecular weight of the polysaccharide polymer was measured using gel permeation chromatography (GPC).

[0175] PSS SECcurity consists of a pump, autosampler, and column oven. 2 GPC experiments were performed using the apparatus. A 300×8mm column with an average particle size of 10μm, SUPREMA LIN XL (PSS Polymer Standards Service GmbH, Mainz, Germany), was used at a flow rate of 1.0 mL / min and a column temperature of 25°C. 0.1M NaNO3 was used as the eluent.

[0176] A sample with a concentration of 1 mg / ml was filtered through a 0.45 μm HA filter before measurement. The injection volume was 50 μL. Detection was performed using an RI detector.

[0177] Data collection and evaluation were performed using PSS WINGPC UniChrom (PSS Polymer Standards Service GmbH, Mainz, Germany). Calibration was performed using dextran standards (molecular weight range from 298,000 g / mol to 180 g / mol) (PSS Polymer Standards Service GmbH, Mainz, Germany).

[0178] Generally, calibration is performed by using appropriate standards. Methods for selecting appropriate standards are known to those skilled in the art.

[0179] In the experimental section of this patent application, refer to “Dextran 40 kg / mol,” “Dextran (...Molecular weight (MW) 40,000 g / mol, 4),” “Dextran...MW 40 kg / mol (4),” “Dextran 4,” or variations thereof. This dextran is commercially available, and its packaging indicates a molecular weight of 40 kg / mol (determined by its intrinsic viscosity). However, when the molecular weight of this dextran is determined according to the method described herein (Chapter 2.8), i.e., using gel permeation chromatography (GPC) as described herein (Chapter 2.8), the following results are obtained: Mn = 15099 g / mol, Mw = 33693 g / mol.

[0180] In the experimental section of this patent application, we refer to “dextran 150 kg / mol,” “dextran···MW150 kg / mol(5),” “dextran 5,” or their variations. This dextran is commercially available, and its packaging indicates a molecular weight of 150 kg / mol (determined by its intrinsic viscosity). However, when the molecular weight of this dextran is determined according to the method described herein (Chapter 2.8), i.e., using gel permeation chromatography (GPC) as described herein (Chapter 2.8), the following results are obtained: Mn = 20312 g / mol, Mw = 156959 g / mol. A bimodal distribution of molecular weight was observed.

[0181] 3 Preparations 3.1 Volume: 5 mL

[0182] [Table 4]

[0183] procedure: I) Dissolve B at approximately 55°C for 30 minutes. II) Melt A at approximately 80°C. III) Heat C to 80°C. Add III) to IV)II) while stirring and emulsify (on a water bath). V) Add IV) to I) while stirring. VI) Gently stir and add D to V). VII) Allow to cool without stirring.

[0184] [Table 5]

[0185] procedure: I) Dissolve B at approximately 55°C for 30 minutes. II) Melt A at approximately 80°C. III) Heat C to 55°C for 30 minutes. IV) Add III) to I) while stirring and emulsify (on a water bath). V) Add II) to IV) while stirring. VI) While stirring, add D to V). VII) Allow to cool without stirring.

[0186] [Table 6]

[0187] procedure: I) Dissolve B at approximately 55°C for 30 minutes. II) Melt A at approximately 80°C. III) Heat C to 80°C. Add III) to IV)II) while stirring and emulsify (on a water bath). V) Add IV) to I) while stirring. VI) Gently stir and add D to V). VII) Allow to cool without stirring.

[0188] 3.2 Volume: 25 mL

[0189] [Table 7]

[0190] procedure: I) Dissolve B at approximately 55°C for 60 minutes. II) Melt A at approximately 80°C. III) Heat C to 80°C. IV) and II) are mixed with III), and then homogenized using a high-shear mixer at 16,000 rpm. V) Cool to approximately 20°C while stirring with a magnetic stirrer. VI) Add V) to I) while stirring. VII) Add D to V) while stirring. VIII) Add E while stirring. IX) Allow to cool without stirring.

[0191] 4 Results 4.1 NMR (GMA functionalization and peptide binding) Polysaccharides of different molecular weights, namely dextran, were functionalized to varying degrees with GMA to obtain biopolymer platforms for further peptide bonding. The degree of GMA modification of the polymers was determined using 1H-NMR analysis with DMSO-d6 as the solvent. The results are summarized in Table 1, showing the percentage of GMA-functionalized repeat units, i.e., glucose units in the case of dextran. The calculation is based on the integral of the vinyl signal (approximately 5.5–6.5 ppm) of GMA bound to the biopolymer compared to the integral of the proton signal (approximately 4–5 ppm) of the glucose repeat unit.

[0192] The degree of dextran-GMA functionalization by peptides was calculated from the integral decrease in the vinyl signal (approximately 5.5–6.5 ppm) in the hybrid. The results are summarized in Table 2.

[0193] 4.2 Rheology 4.2.1 Peptide-dextran hybrids A) Adjustability of G' Peptide-dextran hybrids resulted in hydrogels characterized by the moduloability of G' through the type of grafted peptide, the number of peptide grafts (i.e., the peptide functionalization rate), and the weight % of each peptide-dextran hybrid. This was demonstrated with different peptide grafts on polysaccharides, i.e., the dextran backbone.

[0194] i) Dextran skeleton at 40 kg / mol

[0195] [Table 8]

[0196] ii) 150 kg / mol dextran skeleton

[0197] [Table 9]

[0198] B) The self-repair behavior of 1 wt% peptide-dextran hybrid hydrogels was demonstrated by peptide grafting onto a dextran backbone (H1, H9) at 40 kg / mol or 150 kg / mol.

[0199] [Table 10]

[0200] The H1 and H9 hybrid hydrogel exhibited regenerative behavior after mechanical stress.

[0201] C) pH stability and regeneration behavior of peptide-dextran hybrid hydrogel (40 kg / mol dextran skeleton with peptide graft, 1 wt% hydrogel)

[0202] [Table 11]

[0203] D) Salt stability and regeneration behavior of peptide-dextran hybrids (40 kg / mol or 150 kg / mol dextran backbone with peptide grafts, 1 wt% hydrogel)

[0204] [Table 12]

[0205] E) Temperature stability of peptide-dextran hybrids using analysis of regeneration behavior Samples were prepared according to 1.3ai, and rheological measurements were performed using setting 2.2a in accordance with method 2.2ix.

[0206] Mechanical properties of H1 with respect to temperature (40 kg / mol dextran skeleton with peptide graft, 1 wt% hydrogel) i) Heating from 20°C to 60°C

[0207] [Table 13]

[0208] ii) Cooling from 60°C to 20°C

[0209] [Table 14]

[0210] iii) Regeneration behavior after heating to 60°C and cooling back to 20°C: G' and G'' are given as values ​​for 0.1% strain against temperature.

[0211] [Table 15]

[0212] The regeneration behavior of the peptide-dextran hybrid was maintained even after heating (60°C) and cooling (20°C) cycles.

[0213] 4.2.2 Composite materials A) The possibility of tunable mechanical properties (analysis of G', G'' and regeneration behavior) by preparing different composite materials by adding an unmodified polysaccharide backbone (i.e., dextran) or free peptides. The base material is H1.

[0214] [Table 16]

[0215] The addition of negatively charged self - aggregating peptide (e.g., P5) to a hybrid (e.g., H1) having a positively charged self - aggregating peptide graft was beneficial to the regeneration efficiency of the hydrogel after high - strain conditions. In particular, the complex of the hybrid and dextran was able to maintain a similar regeneration efficiency compared to the hybrid alone, while G’ decreased.

[0216] B) Complexes of peptide - dextran hybrids and other polysaccharides, e.g., H5 with xanthan gum

[0217]

Table 17

[0218]

Table 18

[0219] 4.2.3 Preparations

[0220]

Table (19)

[0221] 4.3 Active substances Vitamin C A homogeneous distribution was observed throughout the transparent hydrogel. The hydrogel could be inverted by a glass - tilt test after gelling overnight at room temperature.

[0222] 4.4 Biodegradability The peptide - dextran hybrid H5 is "readily biodegradable" according to the biodegradation test conducted in accordance with OECD301F.

[0223] 4.5 Cell viability Since cell viability was observed to exceed 70% in all cases, it was found that the hydrogel and its components exhibited excellent cytocompatibility and had no cytotoxic effects on A549 cells within the selected experimental settings.

[0224] [Table 20]

[0225] 4.6 Long-term temperature stability of hydrogels (macroscopic) The gel looked good to the naked eye, even after several weeks and months.

[0226] 5. Further synthesis and characterization of hybrid polymers Functionalization of different polysaccharides with glycidyl methacrylate (GMA): In a typical reaction, polysaccharides (15 g) were placed in a flask under a nitrogen atmosphere, dissolved by adding dry dimethyl sulfoxide (DMSO, 90 mL), and stirred (for about 30 minutes). The mixture was heated to 40°C with stirring. 4-(dimethylamino)pyridine (DMAP, 2.25 g) was separately dissolved in dry DMSO (24 mL). Subsequently, GMA (4.7 or 2.3 mL, as shown in Table 21) was added to the DMAP solution, and the mixture was added to the polysaccharide solution. The reaction was stirred at 40°C for 20 hours. All steps were carried out under a nitrogen atmosphere. The reaction was purified by dialysis (molecular weight cutoff (MWCO) 3.5 kDa) in deionized water for 5 days, which included water changes every 2-3 hours on the first day and twice a day for the remainder of the day. The dissolved product was freeze-dried. The product was obtained as a white solid.

[0227] [Table 21]

[0228] Synthesis of hybrid polymers: Peptide P4 is defined in Chapter 1.2.

[0229] Example 5.1) GMA-functionalized tara gum (tara gum-GMA1, 9.7 mg) was dissolved in 0.1% trifluoroacetic acid (TFA) in water (Milli-Q, 0.3 mL) (A). Separately, tris-(2-carboxyethyl)-phosphine (TCEP, 3.6 mg) was dissolved in 0.1% TFA / H2O (0.32 mL) and added to peptide (P4, 22.0 mg), and the mixture (B) was incubated in a shaker at room temperature for 30 minutes. Next, solution B was added to solution A and stirred overnight (18 h) at 66°C under a nitrogen atmosphere (shaker 1000 rpm). The reaction product was purified by ultrafiltration in 0.1% TFA / H2O at 10,000 rpm and 22-25°C using a Vivaspin tube (5x, MWCO 5 kDa, PES). After freeze-drying for three days, the hybrid (Tara gum-Hybrid 1) was obtained as a white powder.

[0230] Example 5.2) GMA-functionalized guar gum (guar gum-GMA2, 11 mg) was dissolved in 0.1% trifluoroacetic acid (TFA) in water (Milli-Q, 0.22 mL) (A). Separately, tris-(2-carboxyethyl)-phosphine (TCEP, 2.64 mg) was dissolved in 0.1% TFA / H2O (0.32 mL) and added to peptide (P4, 22.0 mg), and the mixture (B) was incubated in a shaker at room temperature for 30 minutes. Next, solution B was added to solution A and stirred overnight (18 h) at 66°C under a nitrogen atmosphere (shaker 1000 rpm). The reaction product was purified by ultrafiltration in 0.1% TFA / H2O at 10,000 rpm and 22-25°C using a Vivaspin tube (5x, MWCO 5 kDa, PES). After freeze-drying for 4 days, the hybrid (guar gum-hybrid 2) was obtained as a white powder.

[0231] Example 5.3)GMA-functionalized guar gum (guar gum-GMA1, 7.4 mg) was dissolved in 0.1% trifluoroacetic acid (TFA) in water (Milli-Q, 0.148 mL) (A). Separately, tris-(2-carboxyethyl)-phosphine (TCEP, 1.77 mg) was dissolved in 0.1% TFA / H2O (0.248 mL) and added to peptide (P4, 14.8 mg), and the mixture (B) was incubated in a shaker at room temperature for 30 minutes. Next, solution B was added to solution A and stirred overnight (18 h) at 66°C under a nitrogen atmosphere (shaker 1000 rpm). The reaction product was purified by ultrafiltration in 0.1% TFA / H2O at 10,000 rpm and 22-25°C using a Vivaspin tube (5x, MWCO 5 kDa, PES). After freeze-drying for 4 days, the hybrid (guar gum-hybrid 1) was obtained as a white powder.

[0232] Example 5.4) GMA-functionalized xanthan gum (xanthan gum-GMA, 5 mg) was dissolved in 0.1% trifluoroacetic acid (TFA) in water (Milli-Q, 0.1 mL) (A). Separately, tris-(2-carboxyethyl)-phosphine (TCEP, 1.2 mg) was dissolved in 0.1% TFA / H2O (0.2 mL) and added to peptide (P4, 10.0 mg), and the mixture (B) was incubated in a shaker at room temperature for 30 minutes. Next, solution B was added to solution A and stirred overnight (18 h) at 66°C under a nitrogen atmosphere (shaker 1000 rpm). The reaction product was purified by ultrafiltration in 0.1% TFA / H2O at 10,000 rpm and 22-25°C using a Vivaspin tube (5x, MWCO 5 kDa, PES). After lyophilization for 4 days, the hybrid (xanthan gum-hybrid) was obtained as a white powder.

[0233] Example 5.5)The GMA-functionalized tara gum (tara gum-GMA2, 10 mg) was dissolved in 0.1% trifluoroacetic acid (TFA) in water (MilliQ, 0.2 mL) (A). Separately, tris-(2-carboxyethyl)-phosphine (TCEP, 2.4 mg) was dissolved in 0.1% TFA / H2O (0.3 mL) and added to the peptide (P4, 20.0 mg), and the mixture (B) was incubated in a shaker at room temperature for 30 minutes. Then, solution B was added to solution A, and the mixture was stirred overnight (18 h) at 66 °C under a nitrogen atmosphere (shaker 1000 rpm). The reaction product was purified by ultrafiltration using a Vivaspin tube (5x, MWCO 5 kDa, PES) at 10,000 rpm in 0.1% TFA / H2O at 22 - 25 °C. After freeze-drying for 4 days, the hybrid (tara gum-hybrid2) was obtained as a white powder.

[0234] Hydrogel preparation: The hybrid was pre-dissolved in an acidic solution, and a hydrogel was obtained by adjusting the pH to approximately pH 7. For example, to obtain a 1 wt% hydrogel, 0.3 mg of the hybrid was dissolved in 15 μL of 0.1% TFA / H2O (pH 2). Subsequently, when 15 μL of 300 mM phosphate buffer pH 9 was added to adjust the pH to neutral (pH 7), gelation started.

[0235] Characterization / Rheology: The hydrogel was prepared directly on a rheology plate (8 mm, plate / plate geometry with a solvent trap), and the following measurements were performed: (α) Time sweep (gelation curve): Oscillatory time sweep measurements were performed at 20 °C with a fixed strain and fixed frequency within the range of 0.05 - 1% strain and 1 - 1.6 Hz frequency. (β) Thixotropy measurement 3: To analyze the strain resistance (oscillatory strain sweep) and self-healing ability of the hydrogel, a continuous combined measurement of an oscillatory strain sweep (0.01 - 1000%) at 20 °C with a fixed frequency of 1 Hz and a subsequent oscillatory type sweep measurement at 20 °C for 300 seconds with a fixed strain of 0.1% and a frequency of 1 Hz was performed.

[0236] In particular, the regenerative capacity of the hydrogel was qualitatively evaluated after the removal of high strain, with a "yes" rating if G' > G'' and a "no" rating if G' < G''.

[0237] The results are shown in Table 22.

[0238] [Table 22]

[0239] As shown in Table 22, the hybrid polymers and hydrogels of the present invention have excellent rheological properties. G' and G'' are significantly increased compared to polysaccharide polymers without peptide chains. Various polysaccharide polymers, including charged and uncharged polysaccharide polymers, can be used.

[0240] Example Composition The following example compositions include any of the hybrid polymers of the present invention, Hybrid H, particularly Hybrid Polymers H1 to H11 (as defined in Table 2) or any of the hybrid polymers listed in Table 22 (Tara Gum Hybrid 1, Tara Gum Hybrid 2, Xanthan Gum Hybrid 1, Guar Gum Hybrid 1, or Guar Gum Hybrid 2). Where used herein, % refers to weight percent based on the total weight of the composition.

[0241] [Table 23]

[0242] [Table 24]

[0243] [Table 25]

[0244] Table 26

[0245] Table 27

[0246] Table 28

[0247] Table 29

[0248] Table 30

[0249] Table 31

[0250] Table 32

[0251] Table 33

[0252] Table 34

[0253] Table 35

[0254] Table 36

[0255] Table 37

[0256] Table 38

[0257] Table 39

[0258] Table 40

[0259] Table 41

[0260] Table 42

[0261] Table 43

[0262] Table 44

[0263] Table 45

[0264] Table 46

[0265] Table 47

[0266] Table 48

[0267] Table 49

Claims

1. below: (a) Polysaccharide polymers; and (b) Peptide chain, A hybrid polymer containing [this component].

2. The hybrid polymer according to claim 1, wherein the polysaccharide polymer is selected from dextran, dextrin, hyaluronic acid, chitosan, xanthan gum, fenugreek gum, tara gum, locust bean gum, carrageenan, guar gum, alginate, agar, agarose, starch, amylose, amylopectin, tragacanth gum, tamarind kernel gum, gum arabic, cherry gum, karaya gum, okra gum, cassia gum, chicle gum, konjac gum, ghati gum, pectin, sclerotium gum, gellan gum, paramylon, paramylam, curdlan, cellulose, diutan gum, inulin, derivatives thereof, and mixtures thereof.

3. The hybrid polymer according to claim 1 or 2, wherein at least 25%, preferably at least 30%, of the amino acids in the peptide chain are identical, preferably selected from hydrophobic amino acids, the hydrophobic amino acid being preferably selected from leucine, isoleucine, phenylalanine, valine, tyrosine, and tryptophan, more preferably selected from leucine, isoleucine, phenylalanine, and valine, even more preferably selected from leucine, isoleucine, and phenylalanine, and particularly preferably the hydrophobic amino acid being isoleucine.

4. The hybrid polymer according to any one of claims 1 to 3, wherein at least 15%, preferably at least 20%, of the amino acids in the peptide chain are identical, preferably selected from charged amino acids, the charged amino acid is preferably selected from lysine, aspartic acid, glutamic acid, arginine, and histidine, more preferably selected from lysine, aspartic acid, and glutamic acid, and particularly preferably the charged amino acid is lysine.

5. The hybrid polymer according to any one of claims 1 to 4, wherein at least 10%, preferably at least 20%, of the amino acids in the peptide chain are selected from amino acids having an amide group in their side chain, and the amino acids having an amide group in their side chain are preferably selected from asparagine, glutamine, and mixtures thereof.

6. The hybrid polymer according to any one of claims 1 to 5, wherein the peptide chain comprises at least one amino acid selected from serine, threonine, and mixtures thereof, preferably at least one serine.

7. The hybrid polymer according to any one of claims 1 to 6, wherein the peptide chain is a depsipeptide chain.

8. The hybrid polymer according to any one of claims 1 to 7, wherein the peptide chain is selected from pea protein, hydrolyzed pea protein, soy protein, hydrolyzed soy protein, broad bean protein, hydrolyzed broad bean protein, chickpea protein, for example, aquafaba, hydrolyzed chickpea protein, wheat protein, hydrolyzed wheat protein, rice protein, hydrolyzed rice protein, whey protein, hydrolyzed whey protein, baobab protein, hydrolyzed baobab protein, collagen protein, hydrolyzed collagen protein, plant collagen-like protein, hydrolyzed plant collagen-like protein, hemp seed protein, hydrolyzed hemp seed protein, jojoba protein, hydrolyzed jojoba protein, keratin protein, hydrolyzed keratin protein, lupine protein, hydrolyzed lupine protein, oat protein, hydrolyzed oat protein, quinoa protein, hydrolyzed quinoa protein, and mixtures thereof.

9. The hybrid polymer according to any one of claims 1 to 8, wherein 1 to 100%, preferably 3 to 50%, more preferably 5 to 40%, even more preferably 8 to 30%, and particularly preferably 10 to 25% of the monosaccharide units of the polysaccharide polymer are optionally modified with the peptide chain through a linker.

10. The hybrid polymer according to any one of claims 1 to 9, wherein the peptide chain is optionally linked to the polysaccharide polymer via a linker through the N-terminal amino acid of the peptide chain.

11. Blends including the following: (a) a hybrid polymer as defined in any one of claims 1 to 10, in an amount of 1 to 99% by weight, preferably 20 to 80% by weight, more preferably 30 to 70% by weight, even more preferably 40 to 65% by weight, and particularly preferably 50 to 60% by weight, based on the total weight of the blend; and (b) 1 to 99% by weight, preferably 20 to 80% by weight, more preferably 30 to 70% by weight, even more preferably 35 to 60% by weight, and especially preferably 40 to 50% by weight, of the total weight of the blend, one or more polysaccharide polymers and / or one or more peptides.

12. Use of a hybrid polymer as defined in any one of claims 1 to 10 or a blend as defined in claim 11 as a rheological modifier.

13. A hydrogel comprising a hybrid polymer as defined in any one of claims 1 to 10 or a blend as defined in claim 11, water, optionally a pH adjuster, and optionally an ionic strength adjuster.

14. The hydrogel according to claim 13, further comprising an active ingredient, wherein the active ingredient is preferably selected from vitamins, humectants, anti-aging agents, anti-wrinkle agents, anti-inflammatory agents, amino acids, peptides, and mixtures thereof.

15. Preparations including the following: (a) 0.1 to 10% by weight, preferably 0.2 to 5% by weight, more preferably 0.3 to 3% by weight, even more preferably 0.4 to 2% by weight, and particularly preferably 0.5 to 1% by weight, of the total weight of the preparation, a hybrid polymer as defined in any one of claims 1 to 10 or a blend as defined in claim 11; and (b) One or more further components in an amount of 90 to 99.9% by weight, preferably 95 to 99.8% by weight, more preferably 97 to 99.7% by weight, even more preferably 98 to 99.6% by weight, and particularly preferably 99 to 99.5% by weight, based on the total weight of the preparation.

16. The preparation according to claim 15, wherein the preparation is a cosmetic preparation, preferably a skin care preparation or a hair care preparation.

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