CN114031668B - Acetyl tetrapeptide-5 analogue with anti-wrinkle effect and moisturizing peptide containing same - Google Patents
Acetyl tetrapeptide-5 analogue with anti-wrinkle effect and moisturizing peptide containing same Download PDFInfo
- Publication number
- CN114031668B CN114031668B CN202210024803.5A CN202210024803A CN114031668B CN 114031668 B CN114031668 B CN 114031668B CN 202210024803 A CN202210024803 A CN 202210024803A CN 114031668 B CN114031668 B CN 114031668B
- Authority
- CN
- China
- Prior art keywords
- moisturizing
- acetyl tetrapeptide
- wrinkle effect
- analogue
- peptide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
- C07K5/0205—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -NH-(X)3-C(=0)-, e.g. statine or derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/37—Esters of carboxylic acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
- A61K8/733—Alginic acid; Salts thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/005—Preparations for sensitive skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/59—Mixtures
- A61K2800/592—Mixtures of compounds complementing their respective functions
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Emergency Medicine (AREA)
- Crystallography & Structural Chemistry (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Cosmetics (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention provides an acetyl tetrapeptide-5 analogue with an anti-wrinkle effect and a moisturizing peptide containing the same, belongs to the field of polypeptide cosmetics, and particularly relates to the moisturizing peptide with the anti-wrinkle effect, which is prepared by taking the acetyl tetrapeptide-5 analogue as a main active substance, and is added with benzyl carbamate modified alginic acid with the weight content of 1.2-4% and 0.8-1.5% of isooctyl ferulate.
Description
Technical Field
The invention belongs to the field of polypeptide cosmetics, and particularly relates to an acetyl tetrapeptide-5 analogue with an anti-wrinkle effect and a moisturizing peptide containing the same.
Background
Aging is a process essential for human beings, and eyes are the parts of the face which are most susceptible to aging and are exposed to the age at the earliest. With aging, the skin of a person gradually loses elasticity, cells age, water is lost, and wrinkles and pouches are formed around the eyes. With the improvement of living standard of people, various cosmetics for preventing wrinkles and removing pouches are developed in succession.
Acetyl tetrapeptide-5, also known as ophthalmic serine peptide, is one of the most widely used cosmetic peptides. The acetyl tetrapeptide-5 can improve microcirculation of eye skin by inhibiting angiotensin converting enzyme, inhibit glycosylation by increasing SOD activity, prevent protein crosslinking, improve skin elasticity, resist wrinkle, reduce permeability of blood vessel, reduce water exudation, and relieve eye edema.
An anti-wrinkle cosmetic related to acetyl tetrapeptide-5 is disclosed, for example, patent CN112494354A discloses a composition containing acetyl tetrapeptide-5 and an eye cream containing the same, which can obviously improve puffiness of pouches, reduce volume of pouches, enable skin around eyes to be firmer, achieve the effect of removing pouches after long-term use, and prevent pouches from forming. With the research and development of acetyl tetrapeptide-5 cosmetics, the demand of acetyl tetrapeptide-5 is increasing year by year, and the current cosmetics containing acetyl tetrapeptide-5 still have certain defects, such as skin permeability, stability and the like, so that the acetyl tetrapeptide-5 cannot meet the requirements of the current cosmetics. Thus, it is necessary to develop a compound having a structure similar to acetyl tetrapeptide-5.
Disclosure of Invention
The invention aims to at least solve one of the technical problems in the prior art, and therefore, the invention provides an acetyl tetrapeptide-5 analogue which has the effects of resisting wrinkles and removing pouches, and has improved stability compared with acetyl tetrapeptide-5; the invention also provides the moisturizing peptide based on the acetyl tetrapeptide-5 analogue, which has the advantages of good skin permeation and absorption effect, high stability and excellent effects of resisting wrinkles and removing pouches.
In order to achieve the above purpose, the technical scheme provided by the invention is as follows.
Subject one, an acetyl tetrapeptide-5 analogue, the structural formula of which is shown in formula (I):
the acetyl tetrapeptide-5 analogs of the present invention are preferably prepared by the following method:
performing esterification reaction on histidine and methanol to obtain a compound 1, and performing acylation reaction on the compound 1 and tyrosine to obtain a compound 2; and then, carrying out acylation reaction on histidine and phthalyl sulphathiazole to obtain a compound 3, carrying out acylation reaction on the compound 3 and a compound 2 to obtain a compound 4, and finally carrying out saponification reaction under an alkaline condition to obtain a target product.
The chemical reaction formula of the acetyl tetrapeptide-5 analogue is as follows:
further, in the chemical reaction formula, the reaction conditions of the steps are as follows: (i) methanol, thionyl chloride and dimethylformamide at the temperature of 70-80 ℃; (ii) tyrosine, 1-hydroxybenzotriazole, dicyclohexylcarbodiimide, acetone/tetrahydrofuran mixed solution and room temperature; (iii) phthalyl sulphathiazole, dicyclohexyl carbodiimide, N-hydroxysuccinimide, acetone/tetrahydrofuran mixed solution and room temperature; (iv) dicyclohexylcarbodiimide, 1-hydroxybenzotriazole, tetrahydrofuran, room temperature; (v) sodium hydroxide and ethanol at the temperature of 70-90 ℃.
Further, the preparation method of the acetyl tetrapeptide-5 analogue specifically comprises the following steps:
step one, mixing histidine with methanol, adding thionyl chloride and dimethylformamide at 0-4 ℃, stirring for 10-30 min, heating to 70-80 ℃, stirring for reaction for 5-8 h, carrying out reduced pressure distillation to remove methanol, and crystallizing with ethanol to obtain a compound 1;
dissolving tyrosine in an acetone/tetrahydrofuran mixed solution, adding 1-hydroxybenzotriazole and dicyclohexylcarbodiimide at 0-4 ℃, stirring for 20-30 min, then heating to room temperature, adding the compound 1, continuously stirring and reacting for 6-10 h, carrying out reduced pressure distillation to remove acetone and tetrahydrofuran, filtering, extracting the filtrate for several times by using a sodium bicarbonate solution and ethyl acetate, washing the extract, and drying to obtain a compound 2;
dissolving histidine in an acetone/tetrahydrofuran mixed solution, adding dicyclohexylcarbodiimide and N-hydroxysuccinimide at 0-4 ℃, stirring for 10-20 min, adding phthalyl sulfathiazole, heating to room temperature, stirring for reaction for at least 8 h, adjusting the pH to 5-6, extracting with ethyl acetate, cooling to 0-4 ℃, refrigerating overnight, crystallizing to separate out a precipitate, filtering, and drying to obtain a compound 3;
dissolving the compound 2 in tetrahydrofuran, adding dicyclohexylcarbodiimide and 1-hydroxybenzotriazole at 0-4 ℃, heating to room temperature, adding the compound 3, stirring for reacting for 8-12 h, removing the solvent through reduced pressure distillation after the reaction is finished, adding a sodium bicarbonate solution and ethyl acetate for extraction for several times, washing and drying the extract, and recrystallizing dichloromethane to obtain a compound 4;
and step five, dissolving the compound 4 in absolute ethyl alcohol, dropwise adding a sodium hydroxide solution, heating to 70-90 ℃ after dropwise adding, stirring for reaction for 0.5-4 h, evaporating the solvent to dryness, dissolving and pulping with ethyl alcohol again, filtering, and distilling under reduced pressure to obtain the target compound.
The invention firstly uses histidine, tyrosine and phthalein sulfathiazole as reactants to carry out acylation reaction to obtain acetyl tetrapeptide-5 analogue, compared with acetyl tetrapeptide-5, the acetyl tetrapeptide-5 analogue replaces serine and N-acetyl-beta-alanine in the structure of the acetyl tetrapeptide-5 by tyrosine and phthalyl sulfathiazole, so that the obtained acetyl tetrapeptide-5 analogue not only enhances the effects of resisting wrinkles and removing pouches, the stability of the acetyl tetrapeptide-5 analogue is improved probably because the sulfonamide group is introduced into the structure of the acetyl tetrapeptide-5 analogue, so that the degradation of the amide group in the acetyl tetrapeptide-5 analogue can be reduced, the low temperature resistance and the high temperature resistance are improved, the acetyl tetrapeptide-5 analogue can be stored at the temperature of-10 to-15 ℃ and the temperature of 40 to 45 ℃, and the stability is improved.
The second subject provides the application of the acetyl tetrapeptide-5 analogue in the preparation of the moisturizing peptide with the anti-wrinkle effect.
The application comprises improving the anti-wrinkle and eye pouch-removing effects of the moisturizing peptide with the anti-wrinkle effect.
The use also includes improving the stability of moisturizing peptides with anti-wrinkle efficacy.
And the third topic is that the moisturizing peptide with the anti-wrinkle effect takes the acetyl tetrapeptide-5 analogue as a main component, and the addition amount of the moisturizing peptide accounts for 3-8% of the weight of the moisturizing peptide.
The moisturizing peptide with the anti-wrinkle effect is further added with benzyl carbamate modified sodium alginate with the weight content of 1.2-4% and isooctyl ferulate with the weight content of 0.8-1.5%.
The weight ratio of the benzyl carbamate modified sodium alginate to the isooctyl ferulate is 2-3: 1.
The preparation method of the benzyl carbamate modified sodium alginate comprises the following steps:
dissolving sodium alginate in deionized water to prepare a solution, adjusting the pH value to 3-4, adding 4-nitrophenyl carbonate while stirring, standing for 8-15 min, adding benzyl carbamate, reacting at 50-70 ℃ for 1.5-5 h, cooling to room temperature after the reaction is finished, adding carbodiimide hydrochloride and N-hydroxysuccinimide, adding ethylenediamine, reacting for 1-4 h, dialyzing, and freeze-drying to obtain the benzyl carbamate modified sodium alginate.
The mass ratio of the sodium alginate to the benzyl carbamate is 1: 2.4-3.2.
The sodium alginate is a natural marine organism polysaccharide extracted from marine brown algae, is a common additive in cosmetics, and has certain antibacterial, anti-inflammatory, anti-wrinkle and anti-aging effects, benzyl carbamate is used for modifying the sodium alginate, the molecular weight of the modified sodium alginate is increased, the water solubility is improved, the modified sodium alginate and the isooctyl ferulic acid are added into the moisturizing peptide with the anti-wrinkle effect, the moisturizing effect of the moisturizing peptide can be obviously improved, and the skin absorption is promoted, which probably is that the acetyl tetrapeptide-5 analogue can be promoted to penetrate through a monolayer cell layer due to the certain synergistic effect of the modified sodium alginate and the isooctyl ferulic acid, the permeation and absorption performance of the skin is improved, and the effect of the moisturizing peptide is increased; in addition, the stability of the moisturizing peptide can be further improved, and the storage stability is improved.
The moisturizing peptide with the anti-wrinkle effect specifically comprises the following components in percentage by weight: 3-8% of acetyl tetrapeptide-5 analogue, 1.2-4% of benzyl carbamate modified sodium alginate, 0.8-1.5% of isooctyl ferulate, 10-20% of glycerol, 10-25% of ethanol, 0.1-5% of auxiliary agent and the balance of deionized water.
The auxiliary agent is any one of an emulsifier, a co-emulsifier, a thickener and a pH regulator.
The moisturizing peptide with the anti-wrinkle effect provided by the invention has the advantages that through the synergistic effect of the components, the stability is high, the skin absorption effect is good, the moisturizing, anti-wrinkle and eye pouch removing effects are realized, wrinkles and eye pouches around eyes can be removed, the generation of wrinkles and eye pouches around the eyes can be prevented, the metabolism of the skin is promoted, the generation of new cells of the skin is stimulated, the skin is tender and glossy, the side effect is small, the safety is high, and the moisturizing peptide can be used for a long time.
The preparation method of the moisturizing peptide with the anti-wrinkle effect comprises the following steps:
1) mixing the auxiliary agents to prepare a phase A;
2) adding acetyl tetrapeptide-5 analogue, benzyl carbamate modified sodium alginate and isooctyl ferulate into deionized water, heating to dissolve, cooling to room temperature, adding glycerol and ethanol, and stirring to obtain phase B;
3) mixing the phase A and the phase B, homogenizing in vacuum, and stirring to obtain the moisturizing peptide with the anti-wrinkle effect.
Further, the heating and mixing temperature in the step 2) is 60-80 ℃.
And fourthly, application of the moisturizing peptide with the anti-wrinkle effect in preparation of eye cream and face cream.
Compared with the prior art, the invention has the technical advantages that:
1) the moisturizing peptide with the anti-wrinkle effect is high in stability, good in skin absorption effect, high in safety and capable of replenishing water, moisturizing, resisting wrinkles and removing pouches, and can be used for a long time;
2) acetyl tetrapeptide-5 analogue is added into the moisturizing peptide as a main active ingredient, so that the effects of resisting wrinkles and removing pouches are enhanced, and the stability is improved compared with that of acetyl tetrapeptide-5;
3) the benzyl carbamate modified sodium alginate and the isooctyl ferulate are added into the moisturizing peptide according to a certain proportion, so that the moisturizing effect of the moisturizing peptide is improved, the permeation and absorption effect on skin is improved, and the stability of the moisturizing peptide can be enhanced.
Drawings
FIG. 1 is a schematic representation of acetyl tetrapeptide-5 analogs of example 113C-NMR chart;
FIG. 2 is a FTIR plot of acetyl tetrapeptide-5 analogs of example 1;
fig. 3 is a graph showing the results of a skin permeation effect test of a moisturizing peptide having an anti-wrinkle effect;
FIG. 4 is a diagram showing the result of a pouch-removing effect test of moisturizing peptides having an anti-wrinkle effect;
fig. 5 is a schematic view showing the anti-wrinkle effect test result of the moisturizing peptide having anti-wrinkle effect.
Detailed Description
In order to make the technical solutions of the present invention more apparent to those skilled in the art, the following examples are given for illustration. It should be noted that the following examples are not intended to limit the scope of the claimed invention.
The starting materials, reagents or apparatuses used in the following examples are conventionally commercially available or can be obtained by conventionally known methods, unless otherwise specified.
Example 1: moisturizing peptide with anti-wrinkle effect
The moisturizing peptide with the anti-wrinkle effect comprises the following components in percentage by weight: 6% of acetyl tetrapeptide-5 analogue, 2.8% of benzyl carbamate modified sodium alginate, 1% of isooctyl ferulate, 14% of glycerol, 18% of ethanol, 1% of glycerol stearate, 1% of citric acid and the balance of deionized water.
The acetyl tetrapeptide-5 analogue is prepared by the following method:
step one, mixing 15.5 g of histidine with 60 g of methanol, adding 0.3 g of thionyl chloride and 0.1 g of dimethylformamide at 0 ℃, stirring for 20 min, heating to 75 ℃, stirring for reaction for 6 h, distilling under reduced pressure to remove methanol, and crystallizing with ethanol to obtain 16.9 g of compound 1;
step two, dissolving 2.5 g of tyrosine in 400 g of acetone/tetrahydrofuran mixed solution with the mass ratio of 2: 1, adding 0.3 g of 1-hydroxybenzotriazole and 0.2 g of dicyclohexylcarbodiimide at 0 ℃, stirring for 25 min, raising the temperature to room temperature, adding the compound 1, continuing stirring for reaction for 8 h, carrying out reduced pressure distillation to remove acetone and tetrahydrofuran, filtering, extracting the filtrate for several times by using sodium bicarbonate solution and ethyl acetate, washing the extract, and drying to obtain 32.9 g of a compound 2;
step three, dissolving 15.5 g of histidine in 230 g of acetone/tetrahydrofuran mixed solution with the mass ratio of 2: 1, adding 0.5 g of dicyclohexylcarbodiimide and 0.2 g N-hydroxysuccinimide at 0 ℃, stirring for 15 min, adding 52.4 g of phthaleinathiazole, heating to room temperature, stirring for reaction for 12 h, adjusting the pH to 5.4, extracting with ethyl acetate, cooling to 0 ℃, refrigerating overnight, crystallizing, precipitating, filtering and drying to obtain 50.3 g of a compound 3;
step four, adding 32.9 g of the compound 2 into 500 g of tetrahydrofuran, adding 0.5 g of dicyclohexylcarbodiimide and 0.3 g of 1-hydroxybenzotriazole at 0 ℃, heating to room temperature, adding 50.3 g of the compound 3, stirring for reaction for 10 hours, removing the solvent by reduced pressure distillation after the reaction is finished, adding a sodium bicarbonate solution and ethyl acetate for extraction for several times, washing the extract, drying, and recrystallizing dichloromethane to obtain 79.5 g of a compound 4;
step five, dissolving 79.5 g of the compound 4 in 800 g of absolute ethyl alcohol, dropwise adding a sodium hydroxide solution with the mass fraction of 10%, heating to 80 ℃ after dropwise adding, stirring for reaction for 3 h, evaporating the solvent, dissolving in ethanol again, pulping, filtering, and distilling under reduced pressure to obtain 74.8 g of acetyl tetrapeptide-5 analogue with the molecular weight of 840.2 measured by GPC.
The benzyl carbamate modified sodium alginate is prepared by the following method:
dissolving 8 g of sodium alginate in 1L of deionized water to prepare a solution, adjusting the pH value to 3.5 by using hydrochloric acid, adding 0.5 g of 4-nitrophenyl carbonate while stirring, standing for 10 min, adding 24 g of benzyl carbamate, placing the mixture in a constant-temperature water bath kettle at 60 ℃, reacting for 4 h, cooling to 30 ℃ after the reaction is finished, adding 0.8 g of carbodiimide hydrochloride and 0.4 g N-hydroxysuccinimide, then adding ethylenediamine, reacting for 4 h, dialyzing, and freeze-drying to obtain the benzyl carbamate modified sodium alginate.
The moisturizing peptide with the anti-wrinkle effect is prepared by the following steps:
1) mixing glyceryl stearate and citric acid to obtain phase A;
2) adding acetyl tetrapeptide-5 analogue, benzyl carbamate modified sodium alginate and isooctyl ferulate into deionized water, heating to dissolve, cooling to room temperature, adding glycerol and ethanol, and stirring to obtain phase B;
3) mixing the phase A and the phase B, homogenizing in vacuum, and stirring to obtain the moisturizing peptide with the anti-wrinkle effect.
Example 2: moisturizing peptide with anti-wrinkle effect
The moisturizing peptide with the anti-wrinkle effect comprises the following components in percentage by weight: 6% of acetyl tetrapeptide-5, 2.8% of benzyl carbamate modified sodium alginate, 1% of isooctyl ferulate, 14% of glycerol, 18% of ethanol, 1% of citric acid and the balance of deionized water.
The preparation method of the benzyl carbamate modified sodium alginate is the same as that in example 1, and the preparation method of the moisturizing peptide is the same as that in example 1.
Example 3: moisturizing peptide with anti-wrinkle effect
The moisturizing peptide with the anti-wrinkle effect comprises the following components in percentage by weight: 6% of acetyl tetrapeptide-5 analogue, 2.8% of sodium alginate, 1% of isooctyl ferulate, 14% of glycerol, 18% of ethanol, 1% of citric acid and the balance of deionized water.
The acetyl tetrapeptide-5 analogue is the same as in example 1, and the preparation method of the moisturizing peptide is the same as in example 1.
Example 4: moisturizing peptide with anti-wrinkle effect
The moisturizing peptide with the anti-wrinkle effect comprises the following components in percentage by weight: 6% of acetyl tetrapeptide-5 analogue, 1.9% of benzyl carbamate modified sodium alginate, 1.9% of isooctyl ferulate, 14% of glycerol, 18% of ethanol, 1% of citric acid and the balance of deionized water.
The acetyl tetrapeptide-5 analogue is the same as in example 1, the benzyl carbamate modified sodium alginate is the same as in example 1, and the preparation method of the moisturizing peptide is the same as in example 1.
Example 5: moisturizing peptide with anti-wrinkle effect
The moisturizing peptide with the anti-wrinkle effect comprises the following components in percentage by weight: 6% of acetyl tetrapeptide-5 analogue, 2.6% of benzyl carbamate modified sodium alginate, 1.3% of isooctyl ferulate, 14% of glycerol, 18% of ethanol, 1% of citric acid and the balance of deionized water.
The acetyl tetrapeptide-5 analogue is the same as in example 1, the benzyl carbamate modified sodium alginate is the same as in example 1, and the preparation method of the moisturizing peptide is the same as in example 1.
Example 6: moisturizing peptide with anti-wrinkle effect
The moisturizing peptide with the anti-wrinkle effect comprises the following components in percentage by weight: 6% of acetyl tetrapeptide-5 analogue, 2.85% of benzyl carbamate modified sodium alginate, 0.95% of isooctyl ferulate, 14% of glycerol, 18% of ethanol, 1% of citric acid and the balance of deionized water.
The acetyl tetrapeptide-5 analogue is the same as in example 1, the benzyl carbamate modified sodium alginate is the same as in example 1, and the preparation method of the moisturizing peptide is the same as in example 1.
Example 7: moisturizing peptide with anti-wrinkle effect
The moisturizing peptide with the anti-wrinkle effect comprises the following components in percentage by weight: 6% of acetyl tetrapeptide-5 analogue, 3.2% of benzyl carbamate modified sodium alginate, 0.8% of isooctyl ferulate, 14% of glycerol, 18% of ethanol, 1% of citric acid and the balance of deionized water.
The acetyl tetrapeptide-5 analogue is the same as in example 1, the benzyl carbamate modified sodium alginate is the same as in example 1, and the preparation method of the moisturizing peptide is the same as in example 1.
Example 8: moisturizing peptide with anti-wrinkle effect
The moisturizing peptide with the anti-wrinkle effect comprises the following components in percentage by weight: 6% of acetyl tetrapeptide-5 analogue, 2.8% of benzyl carbamate modified sodium alginate, 14% of glycerol, 18% of ethanol, 1% of citric acid and the balance of deionized water.
The acetyl tetrapeptide-5 analogue is the same as in example 1, the benzyl carbamate modified sodium alginate is the same as in example 1, and the preparation method of the moisturizing peptide is the same as in example 1.
Example 9: moisturizing peptide with anti-wrinkle effect
The moisturizing peptide with the anti-wrinkle effect comprises the following components in percentage by weight: 6% of acetyl tetrapeptide-5 analogue, 1% of ferulic acid isooctyl ester, 14% of glycerol, 18% of ethanol, 1% of citric acid and the balance of deionized water.
The acetyl tetrapeptide-5 analogue is the same as in example 1, the benzyl carbamate modified sodium alginate is the same as in example 1, and the preparation method of the moisturizing peptide is the same as in example 1.
Example 10: moisturizing peptide with anti-wrinkle effect
The moisturizing peptide with the anti-wrinkle effect comprises the following components in percentage by weight: 6% of acetyl tetrapeptide-5 analogue, 2.8% of benzyl carbamate modified sodium alginate, 1% of isooctyl ferulate, 14% of glycerol, 18% of ethanol, 1% of glycerol stearate, 1% of citric acid and the balance of deionized water.
The acetyl tetrapeptide-5 analogue is prepared by the following method:
step one, mixing 15.5 g of histidine with 60 g of methanol, adding 0.3 g of thionyl chloride and 0.1 g of dimethylformamide at 0 ℃, stirring for 20 min, heating to 75 ℃, stirring for reaction for 6 h, distilling under reduced pressure to remove methanol, and crystallizing with ethanol to obtain 16.9 g of compound 1;
step two, dissolving 2.5 g of tyrosine in 400 g of acetone/tetrahydrofuran mixed solution with the mass ratio of 2: 1, adding 0.3 g of 1-hydroxybenzotriazole and 0.2 g of dicyclohexylcarbodiimide at 0 ℃, stirring for 25 min, raising the temperature to room temperature, adding the compound 1, continuing stirring for reaction for 8 h, carrying out reduced pressure distillation to remove acetone and tetrahydrofuran, filtering, extracting the filtrate for several times by using sodium bicarbonate solution and ethyl acetate, washing the extract, and drying to obtain 32.9 g of a compound 2;
step three, dissolving 15.5 g of histidine in 230 g of acetone/tetrahydrofuran mixed solution with the mass ratio of 2: 1, adding 0.5 g of dicyclohexylcarbodiimide and 0.2 g N-hydroxysuccinimide at 0 ℃, stirring for 15 min, adding 52.4 g of phthaleinathiazole, heating to room temperature, stirring for reaction for 12 h, adjusting the pH to 5.4, extracting with ethyl acetate, cooling to 0 ℃, refrigerating overnight, crystallizing, precipitating, filtering and drying to obtain 50.3 g of a compound 3;
step four, adding 32.9 g of the compound 2 into 500 g of tetrahydrofuran, adding 0.5 g of dicyclohexylcarbodiimide and 0.3 g of 1-hydroxybenzotriazole at 0 ℃, heating to room temperature, adding 50.3 g of the compound 3, stirring for reaction for 10 hours, removing the solvent by reduced pressure distillation after the reaction is finished, adding a sodium bicarbonate solution and ethyl acetate for extraction for several times, washing the extract, drying, and recrystallizing dichloromethane to obtain 79.5 g of a compound 4;
step five, dissolving 79.5 g of the compound 4 in 800 g of absolute ethyl alcohol, dropwise adding a sodium hydroxide solution with the mass fraction of 10%, heating to 80 ℃ after dropwise adding, stirring for reaction for 3 h, evaporating the solvent, dissolving in ethanol again, pulping, filtering, and distilling under reduced pressure to obtain 74.8 g of acetyl tetrapeptide-5 analogue with the molecular weight of 840.2 measured by GPC.
The benzyl carbamate modified sodium alginate is prepared by the following method:
dissolving 8 g of sodium alginate in 1L of deionized water to prepare a solution, adjusting the pH value to 3.5 by using hydrochloric acid, adding 0.5 g of 4-nitrophenyl carbonate while stirring, standing for 10 min, adding 24 g of benzyl carbamate, placing the mixture in a constant-temperature water bath kettle at 60 ℃, reacting for 4 h, cooling to 30 ℃ after the reaction is finished, adding 0.8 g of carbodiimide hydrochloride and 0.4 g N-hydroxysuccinimide, then adding ethylenediamine, reacting for 4 h, dialyzing, and freeze-drying to obtain the benzyl carbamate modified sodium alginate.
The moisturizing peptide with the anti-wrinkle effect is prepared by the following steps:
1) mixing glyceryl stearate and citric acid to obtain phase A;
2) adding acetyl tetrapeptide-5 analogue, benzyl carbamate modified sodium alginate and isooctyl ferulate into deionized water, heating to dissolve, cooling to room temperature, adding glycerol and ethanol, and stirring to obtain phase B;
3) mixing the phase A and the phase B, homogenizing in vacuum, and stirring to obtain the moisturizing peptide with the anti-wrinkle effect.
Experimental example 1: structural characterization of acetyl tetrapeptide-5 analogs
(1) Nuclear magnetic characterization of13C-NMR）
Using nuclear magnetic resonance spectrometer (NMR), with CDCl3As a solvent, to acetyl tetrapeptide-5 analogs13C-NMR characterization. The results are shown in FIG. 1.
From the observation of fig. 1, the peaks near 167.45, 171.28, 173.02 were assigned to the chemical shift of C = O carbon in the amide group, the chemical shift of C = O carbon in the carboxyl group near 174.45 was assigned to histidine, and the chemical shifts of carbon on the thiazole ring in phthalsulothiazole were assigned near 112.01, 136.85, 171.65.
(2) Infrared Spectrum characterization (FTIR)
And performing infrared characterization on the acetyl tetrapeptide-5 analogue by using an infrared spectrometer. The results are shown in FIG. 2.
Observed from FIG. 2, 3407 cm-1、3343 cm-1The vicinity is the telescopic vibration absorption peak of amide N-H, 3090 cm-1Near benzene ring C-H stretching vibration absorption peak, 1720 cm-1Near the peak of absorption of stretching vibration of carboxyl group C = O, 1645 cm-1The vicinity of the peak is 1528 cm, which is the absorption peak of the telescopic vibration of amide C = O-1Near by the plane oscillation peak of thiazole ring C = N and amido H, 1460 cm-11300 cm near the C = C skeleton absorption peak of stretching vibration of benzene ring-1The vicinity is a characteristic peak of S = O in the sulfonamide group.
Experimental example 2: effect test of moisturizing peptide having anti-wrinkle efficacy
(1) Skin penetration effect test:
the moisturizing peptides with the anti-wrinkle effect prepared in examples 1 to 9 were tested by a vertical diffusion experimental method, which specifically includes: fresh pigskin was cut into slices of 2 cm diameter and 1 mm thickness using PBS buffer pH =7.4 (0.27 g KH)2PO4、1.42 g Na2HPO4Adding 8 g of NaCl and 0.2 g of KCl into 800 mL of deionized water, fully stirring and dissolving, then adding concentrated hydrochloric acid to adjust the pH value to 7.4, and finally fixing the volume to 1L), soaking for 12 h, and placing in a vertical diffusion cell for skin permeation experiments, wherein the temperature of the experiments is constant at 36 ℃, and the test time is 0.5 h and 3 h; measuring the concentration of acetyl tetrapeptide-5 analogue or acetyl tetrapeptide-5 in the diffusion cell by ultraviolet spectrophotometer to determine the cumulative diffusion, according to Fick's law of diffusion, the unit cumulative diffusion M (μ g/cm)2) The calculation formula of (a) is shown as follows:
wherein, Cn-the drug concentration measured at n time points; v-sample volume; v0-total diffusion cell volume; S-Total area of skin tested. The test results are shown in FIG. 3.
As can be seen from fig. 3, the unit cumulative diffusion amounts of the moisturizing peptides prepared in examples 1 and 2 are high at 0.5 h and 3 h, and the unit cumulative diffusion amounts of the moisturizing peptides prepared in examples 1 and 2 are slightly different from each other, the unit cumulative diffusion amounts of the moisturizing peptides prepared in examples 3 to 9 are significantly different from each other, and the unit cumulative diffusion amounts of the moisturizing peptides prepared in examples 3 and 8 and 9 are relatively lowest, which indicates that a certain synergistic effect exists between benzyl carbamate-modified sodium alginate and acetyl ferulate, and thus the active ingredient can be promoted to permeate through the skin, and the skin permeation performance can be improved.
(2) Testing the pouch removing effect:
the moisturizing peptides with the anti-wrinkle effect prepared in the examples 1-9 are used as samples, 45 subjects with the age of 25-35 years are organized to carry out the pouch removing effect test, the subjects are randomly divided into 9 groups, 5 persons in each group, and the test conditions are as follows: keeping the temperature at 25 ℃ and the humidity at 50%; after a subject cleans the face with warm water, the test is started after the subject has a rest in a constant-temperature and constant-humidity room for 0.5-1 h, the lower eyelid and the external canthus of the subject are selected, a proper amount of sample is taken to follow the direction of the texture of the skin of the eye, the eye is massaged from the inner side to the outer side slightly, the eye is used once every day, the volume of the eye bag is tested after the eye bag is used for 30 days, the reduction rate of the volume of the eye bag is (the volume of the eye bag before the use-the volume of the eye bag after the use)/the volume of the eye bag before the use multiplied by 100%, the average value of each group is taken, and the test result is shown in figure 4.
As can be seen from fig. 4, when moisturizing peptide 30 d prepared in example 1 was used, the pouch volume was reduced by 41.23%, the pouch was significantly reduced, and the skin around the eyes became tight, and when moisturizing peptide 30 d prepared in example 2 was used, the pouch volume reduction rate was low and the skin relaxation state around the eyes was improved, compared to example 1, indicating that the pouch removing effect of acetyl tetrapeptide-5 analog was better than that of acetyl tetrapeptide-5; from the data of examples 3-9, it can be seen that when only benzyl carbamate modified sodium alginate or isooctyl ferulate is added to the moisturizing peptide (examples 8 and 9), or unmodified sodium alginate is added (example 3), the reduction rate of the pouch volume is lower than that of example 1, which indicates that the addition of benzyl carbamate modified sodium alginate and isooctyl ferulate in a certain proportion to the moisturizing peptide can promote the osmotic absorption of active ingredients, thereby improving the pouch removing effect.
(3) Anti-wrinkle Effect test
Selecting 45 subjects, averagely dividing into 9 groups, taking the moisturizing peptides with the anti-wrinkle effect prepared in examples 1-9 as samples, and using the test parts of the moisturizing peptides as cheeks and canthus on two sides for 1 time in the morning and evening of each day, and cleaning the face; after 30 days of use, the wrinkle average depth value was measured using a MicroSkin II multifunction skin mirror image analysis system, and the wrinkle average depth reduction rate was calculated, and the wrinkle average depth reduction rate = (average wrinkle depth before use-average wrinkle depth after use)/average wrinkle depth before use × 100%, and averaged. The results are shown in FIG. 5.
As can be seen from fig. 5, the average wrinkle depth was reduced by about 14.8% after using the moisturizing peptide 30 d prepared in example 1, and wrinkles were significantly improved, whereas the average wrinkle depth reduction rate was significantly lower than that of example 1 after using the moisturizing peptide 30 d prepared in example 2, which indicates that the anti-wrinkle effect of the acetyl tetrapeptide-5 analogue was significantly improved compared to that of acetyl tetrapeptide-5; the data obtained in the embodiments 3-9 show that the addition of benzyl carbamate modified sodium alginate and isooctyl ferulate in the moisturizing peptide has a certain influence on the anti-wrinkle effect, and when benzyl carbamate modified sodium alginate and isooctyl ferulate are added in the moisturizing peptide in a certain mass ratio, the skin permeation and absorption performance is good, and the anti-wrinkle effect is obviously improved.
Experimental example 3: stability test of moisturizing peptides with anti-wrinkle efficacy
The moisturizing peptides with the anti-wrinkle effect prepared in the examples 1-9 are used as samples, and are respectively placed in a thermostat at 40 ℃ or a refrigerator at-15 ℃ for 24 hours, then the temperature is returned to the room temperature, and whether the oil-water separation phenomenon occurs or not is observed.
Tests show that after the moisturizing peptides prepared in examples 1, 4-7 are placed at 40 ℃ or-15 ℃ for 24 hours, the moisturizing peptides are recovered to room temperature without oil-water separation, and the heat-resistant and cold-resistant stability is high; after the moisturizing peptide prepared in the embodiment 2 is placed at 40 ℃ or-15 ℃ for 24 hours, the moisturizing peptide is restored to room temperature, and an obvious oil-water separation phenomenon appears, which shows that compared with acetyl tetrapeptide-5, N-acetyl-beta-alanine is replaced by sulfanilamide thiazole in the acetyl tetrapeptide-5 analogue, and the heat resistance and cold resistance stability of the moisturizing peptide are improved; after the moisturizing peptides prepared in examples 3, 8 and 9 are placed at 40 ℃ or-15 ℃ for 24 hours, the moisturizing peptides are restored to room temperature, and the phenomenon of oil-water separation is also obvious, and the phenomenon of oil-water separation is more obvious after the moisturizing peptides are placed at-15 ℃, which shows that the stability of the moisturizing peptides can be further improved by simultaneously adding benzyl carbamate modified sodium alginate and isooctyl ferulate in a certain proportion.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Claims (10)
1. An acetyl tetrapeptide-5 analog having the formula:
2. a process for preparing an acetyl tetrapeptide-5 analog according to claim 1, comprising:
performing esterification reaction on histidine and methanol to obtain a compound 1, and performing acylation reaction on the compound 1 and tyrosine to obtain a compound 2; then, performing acylation reaction on histidine and phthalyl sulphathiazole to obtain a compound 3, performing acylation reaction on the compound 3 and a compound 2 to obtain a compound 4, and finally performing saponification reaction under an alkaline condition to obtain a target product;
the chemical reaction formula is as follows:
3. use of an acetyl tetrapeptide-5 analogue according to claim 1 for the preparation of a moisturizing peptide with anti-wrinkle effect.
4. A moisturizing peptide with an anti-wrinkle effect, characterized in that the acetyl tetrapeptide-5 analogue of claim 1 or 2 is used as a main effective component, and the addition amount of the acetyl tetrapeptide-5 analogue is 3-8% of the weight of the moisturizing peptide.
5. The moisturizing peptide with anti-wrinkle effect according to claim 4, further comprising benzyl carbamate-modified sodium alginate in an amount of 1.2-4% by weight and isooctyl ferulate in an amount of 0.8-1.5% by weight.
6. The moisturizing peptide with anti-wrinkle effect as claimed in claim 5, wherein the weight ratio of benzyl carbamate modified sodium alginate to isooctyl ferulate is 2-3: 1.
7. The moisturizing peptide with anti-wrinkle effect as claimed in claim 5, wherein the preparation method of benzyl carbamate modified sodium alginate comprises the following steps:
dissolving sodium alginate in deionized water to prepare a solution, adjusting the pH value to 3-4, adding 4-nitrophenyl carbonate while stirring, standing for 8-15 min, adding benzyl carbamate, reacting at 50-70 ℃ for 1.5-5 h, cooling to room temperature after the reaction is finished, adding carbodiimide hydrochloride and N-hydroxysuccinimide, adding ethylenediamine, reacting for 1-4 h, dialyzing, and freeze-drying to obtain the benzyl carbamate modified sodium alginate.
8. The moisturizing peptide with anti-wrinkle effect according to claim 5, further comprising glycerol, ethanol, an auxiliary agent, deionized water; the auxiliary agent is any one of an emulsifier, a co-emulsifier, a thickener and a pH regulator.
9. The method for preparing the moisturizing peptide with anti-wrinkle effect according to any one of claims 4 to 8, comprising:
1) mixing the auxiliary agents to prepare a phase A;
2) adding acetyl tetrapeptide-5 analogue, benzyl carbamate modified sodium alginate and isooctyl ferulate into deionized water, heating to dissolve, cooling to room temperature, adding glycerol and ethanol, and stirring to obtain phase B;
3) mixing the phase A and the phase B, homogenizing in vacuum, and stirring to obtain the moisturizing peptide with the anti-wrinkle effect.
10. Use of the moisturizing peptide with anti-wrinkle effect according to any one of claims 4 to 8 in the preparation of eye cream and face cream.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210024803.5A CN114031668B (en) | 2022-01-11 | 2022-01-11 | Acetyl tetrapeptide-5 analogue with anti-wrinkle effect and moisturizing peptide containing same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210024803.5A CN114031668B (en) | 2022-01-11 | 2022-01-11 | Acetyl tetrapeptide-5 analogue with anti-wrinkle effect and moisturizing peptide containing same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114031668A CN114031668A (en) | 2022-02-11 |
| CN114031668B true CN114031668B (en) | 2022-04-12 |
Family
ID=80141649
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210024803.5A Active CN114031668B (en) | 2022-01-11 | 2022-01-11 | Acetyl tetrapeptide-5 analogue with anti-wrinkle effect and moisturizing peptide containing same |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114031668B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2907006A1 (en) * | 2006-10-17 | 2008-04-18 | Life Lab Sarl | Cosmetic composition, useful in facial care, comprises allantoin, acetyl tetrapeptide-5 and a mixture of palmitoyl oligopeptide/palmitoyl tetrapeptide-7 |
| CN112494354A (en) * | 2020-12-09 | 2021-03-16 | 珠海为你而美丽化妆品有限公司 | Composition containing acetyl tetrapeptide-5 and eye cream containing composition |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2896646A1 (en) * | 2012-12-27 | 2014-07-03 | Hayashibara Co., Ltd. | Skin-exterior anti-ageing composition and production method therefor |
| CN113786361B (en) * | 2021-09-26 | 2024-01-26 | 湖北省麦诗特生物科技有限公司 | Eye cream composition with pouch removing effect and preparation method thereof |
| CN113893215B (en) * | 2021-10-09 | 2023-08-08 | 广州市涵美化妆品有限公司 | Eye tightening and repairing composition, application and eye cream and preparation method thereof |
-
2022
- 2022-01-11 CN CN202210024803.5A patent/CN114031668B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2907006A1 (en) * | 2006-10-17 | 2008-04-18 | Life Lab Sarl | Cosmetic composition, useful in facial care, comprises allantoin, acetyl tetrapeptide-5 and a mixture of palmitoyl oligopeptide/palmitoyl tetrapeptide-7 |
| CN112494354A (en) * | 2020-12-09 | 2021-03-16 | 珠海为你而美丽化妆品有限公司 | Composition containing acetyl tetrapeptide-5 and eye cream containing composition |
Non-Patent Citations (1)
| Title |
|---|
| 美容肽在化妆品中的功效;施跃英等;《中国化妆品》;20160801;第80-82页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114031668A (en) | 2022-02-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5933525B2 (en) | Indigo extract manufacturing method | |
| CN111110584A (en) | Skin care composition with moisturizing and anti-aging effects and preparation method thereof | |
| CN115583988B (en) | Synthetic peptides, compositions and uses thereof | |
| JP2018162325A (en) | Cosmetic compositions comprising marine plants | |
| CN116120402A (en) | Cyclic peptide and cosmetic composition or medicinal composition and application thereof | |
| CN114031668B (en) | Acetyl tetrapeptide-5 analogue with anti-wrinkle effect and moisturizing peptide containing same | |
| CN110075004A (en) | A kind of medical reparation facial mask of compound biological enzyme and preparation method thereof | |
| CN110075003A (en) | A kind of moisturizing content in freckle cream and preparation method thereof containing beta glucan | |
| CN107308035A (en) | The beautiful skin BB frosts of one kind maintenance | |
| Houston et al. | Influence of temperature upon tricaine methane sulphonate uptake and induction of anesthesia in rainbow trout (Salmo gairdneri) | |
| CN111135118A (en) | Anti-aging polypeptide composition | |
| CN114903810B (en) | Peptide-containing anti-wrinkle composition and application thereof | |
| CN110934772A (en) | Wrinkle-removing and anti-aging composition and application thereof in cosmetics | |
| CN110227050A (en) | A kind of anti-aging maintenance sustained release Essence and the anti-aging Face-protecting mask using its preparation | |
| CN114344191A (en) | Moisturizing mask liquid and mask | |
| CN113730289A (en) | Eye and lip care composition with multiple repairing effects and application thereof | |
| CN113876612A (en) | A facial cream composition containing liposome for increasing skin collagen density and its preparation method | |
| CN117338628B (en) | Skin tightening and anti-wrinkle composition and application thereof | |
| CN112842920A (en) | Amino acid facial cleanser with liquid crystal structure | |
| CN106265119B (en) | Polypeptide firming and moisturizing mask and preparation method thereof | |
| CN115581635B (en) | Face cream with good transdermal absorption effect and preparation method thereof | |
| CN111514060B (en) | Liquid foundation with moisturizing and anti-aging effects | |
| CN114344237B (en) | Plant composite maltotetraose moisturizing repair agent and preparation method thereof | |
| CN115040467B (en) | Composition for relieving and resisting allergy, preparation method and application thereof | |
| KR102523118B1 (en) | Hair care composition containing isobutane |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CP01 | Change in the name or title of a patent holder |
Address after: 518000 floor 5, building 206 plant (Building B, Hengyu Center), No. 19, Dengliang Road, Dengliang community, Nanshan street, Nanshan District, Shenzhen, Guangdong Province Patentee after: Shenzhen Branch of Zhejiang Peptide Biology Co.,Ltd. Address before: 518000 floor 5, building 206 plant (Building B, Hengyu Center), No. 19, Dengliang Road, Dengliang community, Nanshan street, Nanshan District, Shenzhen, Guangdong Province Patentee before: Zhejiang Pai peptide biology Co.,Ltd. Shenzhen Branch |
|
| CP01 | Change in the name or title of a patent holder |