Collagen synthesis promoter containing zinc pyrrolidone carboxylate as its active ingredient

Abstract

Use of zinc pyrrolidone carboxylate as a collagen synthesis promoter.

Description

field of expertise

[0001] The present invention relates to a collagen synthesis promoter comprising zinc as an active ingredient and suitable for preventing wrinkles, and to a composition comprising the same. State of the art

[0002] Many substances have been shown to prevent or improve wrinkles. However, there is no single, completely safe substance that can prevent or improve wrinkles, and many studies are still ongoing. There are various reports on the causes of wrinkles, and dry skin is one such cause. It is a well-known fact that reduced moisture content in the stratum corneum leads to the appearance of fine lines. Therefore, moisturizing components are used to maintain a high level of moisture in the stratum corneum. NMF (natural moisturizing factor) is a well-known moisturizing component found in the skin's own stratum corneum. Furthermore, some amino acids, which are constituent building blocks of the stratum corneum, have also been shown to maintain a high level of moisture in the stratum corneum.Furthermore, the prevention and improvement of wrinkles based on promoting the turnover of cutaneous keratinocytes with cell-activating components, such as placenta extract, vitamins and the like, has also been proposed.

[0003] In addition to dryness, a decrease in the amount of collagen in the dermis is considered a major cause of wrinkles and sagging skin associated with skin aging. Collagen in the skin is a component of the dermis's extracellular matrix and plays a vital role as connective tissue. The amount of collagen is an important factor in ensuring the skin's softness and firmness.

[0004] To increase the amount of collagen in skin tissue, it is important to inhibit UV-irradiated collagenase activity, suppress collagen degradation, and promote collagen synthesis. Collagen synthesis is known to be increased by the effects of vitamins, hormones, cytokines, and similar substances.

[0005] Products intended to promote collagen synthesis, such as collagen protein, collagen-forming amino acid compositions, and similar substances, are commercially available. However, their collagen synthesis-promoting effect has not been proven.

[0006] Furthermore, while retinoids, retinol, and retinoic acid are known to promote collagen synthesis, teratogenicity and side effects have also been described, and therefore there is a need for a safe and highly effective collagen synthesis promoter.

[0007] Ubiquinone is a compound discovered in 1957 and isolated from the intracellular mitochondria of bovine heart. It is also known as coenzyme Q10 or simply CoQ10. Ubiquinone is produced in the body and is found in many organs. One of its functions is energy production in the mitochondria. In the mitochondria, a high-energy molecule called ATP is produced using sugars, lipids, and other substances via the electron transport system and the tricarboxylic acid cycle. In other words, ubiquinone acts in the electron transport system and contributes to ATP production. It is said that the amount of ubiquinone produced in the heart decreases to about half the level at age 20 after the age of 40. Regarding the skin, there is limited detailed data available on ubiquinone. However, as with other organs, it is assumed that ubiquinone concentrations decrease with age.

[0008] Another effect of ubiquinone is its antioxidant effect. Since the skin is frequently exposed to ultraviolet rays and similar substances, an oxidation reaction is constantly induced within it. It is believed that many ubiquinones are produced and used to prevent this oxidation reaction. In 2004, ubiquinone became available for cosmetic use and has since been incorporated into numerous cosmetic products. However, aside from promoting skin cell turnover by boosting energy production and its antioxidant effect, many aspects of ubiquinone's specific effects on the skin remain unclear.

[0009] On the other hand, zinc has been reported to suppress diaper rash, as zinc oxide, a type of inorganic zinc salt, is known to do so (e.g., FEBS Letters, Vol. 384, pages 92-96, 1996). However, the compound is sparingly soluble in water and oily components, and therefore difficult to use as a component in cosmetics and topical skin preparations.

[0010] Zinc glycinate, a type of organic zinc salt, is known to promote the induction of metallothionein (JP-A-2005-247729). This has a suppressive effect on skin damage caused by UV radiation, such as inhibiting melanin synthesis, preventing cytotoxicity, and similar effects, achieved through antioxidant activity, free radical scavenging, and the like. The effect of zinc glycinate on collagen synthesis is unknown.

[0011] Zinc pyrrolidone carboxylate and the zinc salt of an acylamino acid, which are also a type of organic zinc salt, are known to suppress UV-induced inflammatory factors such as AP-1, NF-κB, and the like (JP-A-2006-022090, WO2005 / 123062). However, the effect of zinc pyrrolidone carboxylate and the zinc salt of an acylamino acid on collagen synthesis is unknown.

[0012] It is known that other organic and inorganic zinc salts are added to cosmetic formulations to provide astringent, anti-inflammatory, whitening, UV protection, deodorant, and antimicrobial effects. However, using them near the concentration range where an effect is expected is problematic in cosmetics due to the risk of skin irritation. Furthermore, since the aforementioned effects on the skin are absent at low concentrations, zinc salts are not actively used in cosmetics.

[0013] DE102004039550 A1 describes a cosmetic or dermatological topical composition containing (2-hydroxyethyl)urea in a suitable cosmetic or dermatological carrier and at least one active ingredient selected from: a) monomers, oligomers and polymers of amino acids, NC 2-C 24-acyl amino acids, the esters and / or the physiologically acceptable metal salts of these substances, b) DNA or RNA oligonucleotides, c) natural betaine compounds, d) vitamins, provitamins and vitamin precursors of groups A, B, C, E, H and K and the esters of the aforementioned substances, e) α-hydroxycarboxylic acids, α-ketocarboxylic acids, β--Hydroxycarboxylic acids and their ester, lactone or salt form, f) flavonoids and flavonoid-rich plant extracts, g) isoflavonoids and isoflavonoid-rich plant extracts, h) polyphenols and polyphenol-rich plant extracts, i) ubiquinone and ubiquinol and their derivatives, j) silymarin, k) naturally occurring xanthine derivatives selected from caffeine, theophylline, theobromine and aminophylline, i) ectoine, m) inorganic and organic UV filter substances, n) self-tanning agents, o) skin-lightening agents, p) skin-soothing agents, q) sebum-regulating agents, r) and polysaccharides containing deoxy sugars or deoxy sugar building blocks.;

[0014] EP1640041 A2 describes a cosmetic or dermatological composition for topical treatment of the skin, containing in a suitable cosmetic or dermatological carrier a) at least one active ingredient that stimulates collagen synthesis and b) at least one active ingredient that increases and / or improves the interaction between the extracellular matrix and the fibroblasts.

[0015] US20050271606 A1 describes a method for treating or preventing inflammation, comprising the administration of a composition that includes zinc pyrrolidone carboxylate. Disclosure of the invention

[0016] In an attempt to improve wrinkles through collagen synthesis rather than by providing moisture or promoting skin turnover, the inventors conducted intensive studies and found that introducing zinc into human dermal fibroblasts promotes the expression of an ascorbic acid transporter, while simultaneously achieving a collagen synthesis-promoting effect.

[0017] The ascorbic acid transporter is a membrane protein that promotes the uptake of ascorbic acid by a cell (see Hiroyasu Tsukaguchi et al. (1999), Nature 399, 70-75). Increased expression of an ascorbic acid transporter enhances the intracellular uptake of ascorbic acid by dermal fibroblasts, which is essential for collagen synthesis, and promotes collagen synthesis.

[0018] Furthermore, they surprisingly discovered that the addition of zinc pyrrolidone carboxylate, a type of zinc salt, to ubiquinone, which has a conventionally unknown collagen synthesis-promoting effect, produces a highly remarkable synergistic boost to collagen synthesis.

[0019] One objective of the present invention is to provide, based on the above-mentioned result, a collagen synthesis promoter with a superior effect on collagen synthesis and a cosmetic composition containing the same. Means to solve the problems

[0020] Accordingly, the present invention relates to a composition for promoting collagen synthesis comprising zinc as an active ingredient, and to a composition for promoting collagen synthesis comprising ubiquinone and zinc pyrrolidone carboxylate as active ingredients.

[0021] With regard to skin irritation, the efficiency of intracellular introduction, and the like, a truly practical method for introducing zinc requires the conversion of zinc into a zinc salt and its application to the skin in a given concentration, which in particular includes the following invention. (1) Use of zinc pyrrolidone carboxylate as a collagen synthesis promoter. (2) Use of zinc pyrrolidone carboxylate and ubiquinone as collagen synthesis promoters. (3) Use according to claim (2), wherein ubiquinone and zinc pyrrolidone carboxylate are contained in a weight ratio of 50:1 to 0.1:1. (4) Use of a composition to promote collagen synthesis comprising zinc pyrolidone carboxylate and ubiquinone as active ingredients. (5) Use according to claim (4), comprising zinc pyrrolidone carboxylate in a concentration of 1 µM to 100 mM in the composition. (6) Use of a composition to promote collagen synthesis comprising 0.0001 to 10 wt% ubiquinone and 0.000035 to 3.5 wt% zinc pyrrolidone carboxylate. (7) Use of a composition according to one or more of claims 4 to 6, further comprising an N-acylic acid amino acid ester or an amino acid derivative. (8) Use of a composition according to one or more of claims 4 to 7, wherein the composition is an ointment, skin toner, lotion, cream, emulsion, serum, face mask, foundation, liquid hand soap, shampoo or hair lotion. (9) Use of a composition according to one or more of claims 4 to 8, wherein collagen is synthesized via the enhanced expression of an ascorbic acid transporter in human fibroblasts. Brief description of the drawings Fig. 1 confirms the collagen synthesis-promoting effect of a zinc salt using human dermal fibroblasts. Fig.2 confirms the collagen synthesis-promoting effect of zinc pyrrolidone carboxylate using human dermal fibroblasts. Fig. 3 confirms the enhancing effect of zinc pyrrolidone carboxylate on the expression of ascorbic acid transporter mRNA. Fig. 4 confirms the collagen synthesis-promoting effect of zinc pyrrolidone carboxylate and ubiquinone using human dermal fibroblasts. Fig. 5 confirms the collagen synthesis-promoting effect of a preparation containing zinc pyrrolidone carboxylate in three-dimensionally cultured human skin. Detailed description of the invention; Best mode for carrying out the invention

[0022] The present invention is explained in detail below.

[0023] The zinc to be used in the present invention requires, with regard to skin irritation, the efficiency of intracellular delivery, and the like, the actual conversion of zinc into a zinc salt and its application to the skin in a given concentration. In this case, either organic or inorganic zinc salts can be used. Organic zinc salts can include zinc pyrrolidone carboxylate, zinc gluconate, zinc phenolsulfonate, zinc lactate, zinc salts of various amino acids, a zinc salt of acylamino acid, and the like. Inorganic zinc salts can include zinc sulfate, zinc chloride, zinc oxide, and the like.

[0024] Examples of zinc salts of an amino acid include zinc salts of various amino acids, such as zinc glycinate, zinc alaninate, zinc valinate, zinc leucineate, zinc isoleucine, zinc phenylalaninate, zinc methioninate, zinc tryptophanate, zinc asparaginate, zinc glutamate, zinc serine, zinc threoninate, zinc cysteineate, zinc tyrosinate, zinc aspartate, zinc glutamate, zinc lysinate, zinc argininate, zinc histidine, and the like.

[0025] The zinc salt of an acylamino acid is a zinc salt of a compound in which an acyl group is bonded to an amino group of an amino acid. An example of an acylamino acid is a compound with the following structural formula:

[0026] In the above-mentioned formula (I), R1 is an acyl group with 2 to 22 carbon atoms, R2 is a hydrogen atom or a straight-chain or branched alkyl group with 1 to 6 carbon atoms, R3 is a side chain of valine, leucine, isoleucine, phenylalanine, methionine, tryptophan, asparagine, glutamine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamic acid, lysine, arginine, histidine or alanine, which is an amino acid, or a hydrogen atom, and n is an integer of 0 or 1.

[0027] From the perspective of its effect in terms of promoting increased collagen synthesis, the following applies: Since zinc pyrrolidone carboxylate, which has a blood flow-promoting effect, is expected to transport beneficial substances, such as ascorbic acid and the like, in the blood vessels to the application site, it causes a more noticeable influence of the effect promoting the expression of the ascorbic acid transporter on the collagen synthesis-promoting effect.

[0028] Zinc pyrrolidone carboxylate (hereinafter also referred to as PCA zinc salt or PCA-Zn salt) is a zinc salt of 2-pyrrolidone-5-carboxylic acid and can exist in the form of various hydrates. The zinc pyrrolidone carboxylate to be used in the examples is a dihydrate and has the following structure.

[0029] The zinc pyrrolidone carboxylate to be used can be the D-form, L-form, and / or DL-form (a mixture of the D-form and L-form). For example, an L-PCA zinc salt and a DL-PCA zinc salt can be used. These can be used alone or in a mixture. When the DL-form is used, there is no particular restriction on the ratio of D-form to L-form.

[0030] The collagen synthesis promoter of the present invention is used by adding it to a composition, for example, a cosmetic, a topical skin preparation, and the like. In this case, it is important to adjust the amount to be added to a very small quantity of about 1 µM to 100 mM (0.000035 wt.% to 3.5 wt.% as PCA zinc salt), preferably 1 µM to 10 mM (0.000035 wt.% to 0.35 wt.% as PCA zinc salt), and particularly preferably 1 µM to 1 mM (0.000035 wt.% to 0.035 wt.% as PCA zinc salt). An amount of not less than 3.5 wt.% causes problems with regard to skin feel, skin irritation, or the development of a feeling of friction, and the like. An experiment using cultured skin cells shows that, adversely, a collagen synthesis-promoting effect does not occur at high concentrations.

[0031] When the collagen synthesis promoter of the present invention is added to a composition, for example, a cosmetic, a skin preparation for external use, and the like, a component that is generally used as a cosmetic or skin preparation for external use may also be added alongside these components, as long as the effect of the present invention is not impaired.

[0032] A second embodiment of the present invention relates to a collagen synthesis promoter comprising ubiquinone and zinc pyrrolidone carboxylate as active ingredients. As explained above, zinc pyrrolidone carboxylate can be used alone as a collagen synthesis promoter. When used in combination with ubiquinone, it can act as a collagen synthesis promoter with a surprising synergistic effect.

[0033] As a result, an improvement in wrinkles can be expected through collagen synthesis rather than through the promotion of skin turnover or an antioxidant effect.

[0034] In the present invention, "ubiquinone" means ubiquinones in which the number of isoprene units in the side chain of 2,3-dimethoxy-5-methyl-6-(poly)prenyl-1,4-benzoquinone is 10. For the effect of the present invention to be appropriately realized, the amount of ubiquinone added is preferably not less than 0.0001 wt.%, particularly preferably not less than 0.01 wt.%, and most preferably not less than 0.03 wt.%. If the amount of ubiquinone added is unnecessarily high, crystallization and yellowing of the product may occur. Therefore, it is preferably not more than 10 wt.%, particularly preferably not more than 1 wt.%, and most preferably not more than 0.1 wt.%. In order for the effect of the zinc pyrrolidone carboxylate to be appropriately effective, the mixing ratio of ubiquinone and zinc pyrrolidone carboxylate is preferably 50:1 to 0.1:1.

[0035] Another embodiment of the present invention relates to a composition comprising simultaneously the collagen synthesis promoter of the present invention and one or more species selected from the group consisting of N-acyl amino acid esters, amino acids and an amino acid derivative.

[0036] The N-acyl amino acid ester that can be added to the composition of the present invention is represented by the following structure: wherein X and Y can be the same or different and each is at least one species selected from a sterol ester-forming residue, a straight-chain or branched liquid higher alkyl alcohol or alkenyl alcohol having 8 to 30 carbon atoms, and a solid, straight-chain or branched higher alcohol ester-forming residue having 12 to 38 carbon atoms, the COR bonded to a nitrogen atom being a straight-chain acyl group having 8 to 22 carbon atoms, n = 1 or 2, a and b = 0 to 10, and b = 0 to 10.

[0037] In particular, an N-acyl-amino acid diester is preferred, and specific preferred examples include N-lauroylglutamic acid di(chole-sterylbehenyloctyldodecyl), N-lauroylglutamic acid di(cholesteryloctyldodecyl), N-lauroylglutamic acid di(phytosteryl-2-octyldodecyl), N-lauroylglutamic acid di(octyldodecylphytosterylbehenyl) and the like, which are commercially available from Ajinomoto Co., Inc. under the trade names “Eldew CL-301”, “Eldew CL-202”, “Eldew PS-203” and “Eldew PS-304”, respectively.

[0038] In the present invention, the N-acyl amino acid ester also contributes to improving the moisturizing properties and skin texture, as well as to improving stability. To ensure that this effect is appropriately achieved, the amount added is preferably not less than 0.01 wt.%, particularly preferably not less than 0.1 wt.%, and most preferably not less than 0.5 wt.%. If the amount of the N-acyl amino acid ester added is unnecessarily high, stickiness may develop. Therefore, it is preferably not more than 20 wt.%, particularly preferably not more than 10 wt.%, and most preferably not more than 5 wt.%.

[0039] The amino acid or amino acid derivative is added to a composition to enhance its moisturizing properties. Examples of amino acids include alanine, arginine, asparagine, aspartic acid, citrulline, cysteine, cystine, glutamine, glutamic acid, glycine, histidine, oxyproline, isoleucine, leucine, lysine, methionine, ornithine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, and the like. The L-form, D-form, and / or DL-form, or a salt thereof, may be used. Examples of such a salt include a sodium salt, potassium salt, triethanolamine salt, and the like.

[0040] The amino acid derivative is the aforementioned amino acid that has undergone cyclization, acylation, or esterification. Examples include acetylglutamic acid, acetylmethionine, acetylcysteine, N,N-diacetyl-L-cystin dimethyl ester, acylglutamate, acylglycinate, acylalaninate, acylarginine ethyl ester, and the like. However, N-acyl amino acid esters are not included.

[0041] Examples of components commonly used in cosmetics or topical skin preparations include antioxidants, anti-inflammatories, UV absorbers, brighteners, cell stimulants, moisturizers, chelating agents, oily starting materials, surfactants, solvents, polymers, powdered substances, pigments, flavorings, transdermal absorption promoters, steroid hormones, and the like.

[0042] The form of the composition containing the aforementioned zinc salt is not subject to any particular restrictions and can be any form such as a liquid, paste, gel, solid, powder, and the like. Furthermore, the composition of the present invention can be used for oils, lotions, creams, emulsions, gels, shampoos, hair rinses, hair conditioners, varnishes, foundations, lipsticks, face powders, face masks, ointments, tablets, injectable solutions, granules, capsules, perfumes, powders, colognes, toothpastes, soaps, aerosols, cleansing foams, and the like, as well as for anti-aging agents, dermatitis agents, bath preparations, hair growth agents, skin serums, sunscreens, agents for photosensitivity (e.g., xeroderma pigmentosum, photodermatosis, etc.), agents for photoallergy, agents for optical immunosuppression, and agents for skin damage (e.g., injury, chapping, cracks, etc.).), antiseptic, antibacterial agent, insecticide, pesticide, keratolytic, exfoliating agent, acne treatment, treatment for various skin diseases (e.g., keratosis, xeroderma, ichthyosis, psoriasis, etc.) and the like.

[0043] Furthermore, other common components may also be added to the composition, provided that the effect of the present invention is not impaired. Examples of other common components in the composition of the present invention are preservatives, anti-bleaching agents, buffers, acne medications, anti-dandruff / anti-itch agents, adiaphoretic deodorant, burn medications, mite and lice treatments, corneal softeners, xeroderma medications, antiviral agents, hormones, vitamins, amino acids and peptides, proteins, astringents, cooling agents and stimulants, animal / plant components, antibiotics, fungicides, hair growth agents, and the like. Examples

[0044] The present invention is explained in more detail below with reference to the examples (synthesis examples, experimental examples, and preparation examples), which are not to be considered limiting. In these examples, the amount to be added is given in wt.%. Experimental Example 1: Collagen synthesis-promoting effect of zinc salt

[0045] To confirm the collagen synthesis-promoting effect of zinc salt, two types of zinc salt and two types of sodium salt were prepared, and their collagen synthesis-promoting effect on normal human dermal fibroblasts (NHDF) was evaluated. Confluent normal human dermal fibroblasts were detached, spread in a 96-well plate in DMEM (Dulbecco's modified Eagle medium containing 10% FBS), and cultured aseptically for 24 h under saturated steam at 37 °C and 5% CO₂. After 24 h, the medium was removed, and DMEM containing a sample adjusted to a predetermined concentration (50 µM, with 250 µM sodium ascorbate and 2% FBS) was added. The cells were cultured aseptically for 48 h under saturated steam at 37 °C and 5% CO₂.After 48 hours, the culture supernatant was collected, and the amount of procollagen type I released into the liquid by the human dermal fibroblasts was quantitatively assessed by ELISA.

[0046] The results are in Fig. Figure 1 shows that, as the graph illustrates, both types of zinc salts increased the amount of synthesized collagen in the medium over 48 hours compared to the control. However, the corresponding sodium salt did not increase the amount of synthesized collagen in the medium. Experimental Example 2. Blood flow-enhancing effect of PCA-Zn salt

[0047] The difference between the PCA zinc salt and other zinc salts was confirmed based on their blood flow-enhancing effect (increased blood volume). A zinc salt and a sodium salt of each sample were prepared, applied to the human forearm, and the blood flow-enhancing effect was visually assessed. The samples used were (1) PCA zinc salt, (2) PCA sodium salt, (3) zinc gluconate, (4) sodium gluconate, (5) zinc sulfate, and (6) disodium sulfate. The concentration was 5% by weight, and the pH was adjusted to 5.5–6.5. The above-mentioned samples (80 µl) were applied to the forearm in the form of a semi-closed patch. After 30 minutes, the patch was removed and visually assessed. Visual observation and scoring were also performed after 30 and 50 minutes according to Table 1. The results are shown in Table 2. The results show the mean score obtained. Table 1 Points 0 no change 1 very weak promotion of blood flow in part of the application site 2 weak promotion of blood flow in part of the application site 3 weak promotion of blood flow at the application site 4 Promoting blood flow at the application site Table 2 sample 0 minutes after removal of the plaster 30 minutes after removing the plaster 50 minutes after removal of the plaster 5 wt% PCA-2-Zn salt 2,5 2 1 5 wt% PCA sodium salt 2,5 2,5 1 5% by weight zinc gluconate 0 0 0 5% w / w sodium gluconate 0 0 0 5 wt% zinc sulfate 0 0 0 5 wt% disodium sulfate 0 0 0

[0048] Samples (1) and (2) showed a temporary increase in blood flow. However, no increase in blood flow was observed in any of the other samples during the evaluation period. The results indicated that PCA-Zn salt, when applied to the skin, induces a temporary increase in blood flow compared to the other Zn salts.

[0049] Therefore, the PCA-Zn salt is expected to enhance the collagen production-promoting effect more than the other Zn salts, as it transports beneficial substances, such as ascorbic acid and the like, in the blood vessels to the application site.

[0050] Experimental Example 3. Collagen synthesis-promoting effect of zinc pyrrolidone carboxylate on human dermal fibroblasts

[0051] Normal human dermal fibroblasts were spread in a 6-well plate in DMEM (Dulbecco's modified Eagle medium containing 10% FBS). The cells were cultured for 24 h under saturated steam at 37°C and 5% CO2. After 24 h, the medium in the 6-well plate was replaced with DMEM (containing 10% FBS and 500 µM ascorbic acid) spiked with a sample (PCA zinc salt, etc.) at a concentration of 10 µM. The sample was exchanged daily for a total of five times (8 days). 24 h after the last sample exchange, the medium was aspirated, and the well was rinsed with PBS. The resulting collagen plate was enzymatically degraded and dissolved, and the amount of type I collagen was quantitatively assessed by ELISA.

[0052] As in Fig.Figure 2 shows that the amount of collagen type I increased by 180% upon addition of zinc pyrrolidone carboxylate (10 µM) compared to the control (without addition of zinc pyrrolidone carboxylate). Experimental Example 4. The expression of ascorbic acid transporter mRNA enhanced by zinc pyrrolidone carboxylate

[0053] Normal human dermal fibroblasts were spread in a Petri dish in DMEM (Dulbecco's modified Eagle medium containing 10% FBS). The cells were cultured for 1 day under saturated steam at 37 °C and 5% CO2. A sample (PCA zinc salt, etc.) was added to the Petri dish at a concentration of 1 µM or 50 µM, and the cells were cultured in an incubator for 24 h. After culture, the medium was aspirated, and the dish was rinsed with PBS. RNA in the cells was extracted according to the instructions for the RNeasy Mini Kit (manufactured by Qiagen). From the obtained RNA, cDNA was synthesized using a conventional method, and an RT-PCR of the ascorbic acid transporter (SVCT) was performed using a method known from the literature (Christopher P. Corpe et al. (2005), J. Biol. Chem. 280, 5211-5220) (Primer: Sense primer: 5'-CTGAGCTCATGGCGATC-TAC-3' (SEQ ID No. 1), Antisense primer: 5'-CATGTCAGGTAGTGCTGTAGCCCCA-3' (SEQ ID No. 2)).The recovered DNA was subjected to electrophoresis, and the expression level of SVCT RNA was assessed according to the presence or absence of the sample. The electrophoretic band was evaluated using a Lumino Image Analyzer (LAS-3000 / manufactured by Fujifilm Corporation) and numerical values ​​standardized with G3PDH.

[0054] As in Fig. As shown in Figure 3, it was confirmed that the addition of zinc pyrrolidone carboxylate (1 µM, 50 µM) increased the expression level of the ascorbic acid transporter RNA by 150% with the addition of 1 µM and by 200% with the addition of 50 µM compared to the control without addition. This suggests that the ascorbic acid transporter in NHDF increased, which in turn increased the intracellular ascorbic acid concentration, which in turn increased the amount of collagen synthesized in NHDF. Experimental example 5. Collagen synthesis-promoting effect of zinc pyrrolidone carboxylate and ubiquinone on human dermal fibroblasts

[0055] Normal human dermal fibroblasts were spread in a 6-well plate in DMEM (Dulbecco's modified Eagle medium containing 10% FBS). The cells were cultured for 24 h in an incubator under saturated steam at 37°C and 5% CO2. After 24 h, the medium in the 6-well plate was replaced with DMEM (containing 10% FBS and 500 µM ascorbic acid) containing sample (1) PCA zinc salt (30 µM) + ubiquinone (30 µM), sample (2) PCA zinc salt (30 µM), and sample (3) ubiquinone (30 µM). The sample was exchanged daily for a total of five times (8 days). 24 h after the last sample exchange, the medium was aspirated, and the well was rinsed with PBS. The obtained collagen plate was enzymatically decomposed and dissolved, and the amount of collagen type I was quantitatively assessed by ELISA.

[0056] As in Fig. As shown in Figure 4, zinc pyrrolidone carboxylate alone exhibited approximately 300% of the collagen matrix production-promoting effect, and ubiquinone alone showed approximately 250% of the promoting effect compared to the control (without the addition of a sample). When zinc pyrrolidone carboxylate and ubiquinone were administered in combination, the promotion of collagen synthesis increased to approximately 400%, and a stronger collagen synthesis-promoting effect than with zinc pyrrolidone carboxylate alone was confirmed.

[0057] This revealed that a previously unknown combination of ubiquinone and zinc pyrrolidone carboxylate had a collagen synthesis-promoting effect. Since zinc pyrrolidone carboxylate linked the energy-production-enhancing effect of ubiquinone with the promotion of collagen synthesis, the simultaneous use of the two successfully led to a synergistic increase in the promotion of collagen synthesis. Experimental example 6. Collagen synthesis-promoting effect in three-dimensionally cultured human skin by a preparation containing zinc pyrrolidone carboxylate

[0058] Using a three-dimensional cultured model of human skin, consisting of a stratum corneum, an epidermis, and a dermis, and forming a collagen matrix, 100 µl of Cream A, containing PCA zinc salt (0.001 wt%), ubiquinone, N,N'-diacetylcysteine ​​dimethyl ester (DACDM), and several types of amino acids (see Tables 7 and 8), was applied to the surface of a stratum corneum. For comparison, 100 µl of Cream B, which did not contain four of the components of Cream A (PCA zinc salt, ubiquinone, DACDM, and the amino acid group), was applied, and as a control, 100 µl of phosphate buffer (PBS) was applied to the surface of a stratum corneum in the same manner. The dermis side of the skin model was filled with a skin model medium, and the medium was changed daily. After application of the sample, the skin model was cultured in an incubator under saturated steam at 37°C and 5% CO2.Three days after sample application, the upper layer of the sample and the medium of the lower layer were removed, and the skin was rinsed with water and freeze-dried. After freeze-drying, the dermis layer alone was immersed in water for one day. After one day, the procedure used in Experimental Example 2 above was applied to the infiltrated dermis layer, the collagen matrix was enzymatically degraded and dissolved, and the amount of type I collagen was quantitatively assessed by ELISA.

[0059] As in Fig. As shown in Figure 5, in a three-dimensional cultured human skin model to which cream A containing zinc pyrrolidone carboxylate was applied, the amount of type I collagen of the collagen matrix in the dermis layer increased by about 170% compared to the control and to cream B. Synthesis example 1. Synthesis of zinc pyrrolidone carboxylate

[0060] A DL-PCA zinc salt was synthesized according to the procedure described in JP-A-3-168240 by reacting DL-PCA with zinc oxide in water at 100 °C for 2 h, stirring the mixture at room temperature for 5 h, and recovering the precipitated crystals by filtration. An L-PCA zinc salt was synthesized according to the following procedure. An aqueous solution of sodium L-glutamate monohydrate (61.1 g) was heated in an autoclave at 180 °C for 2 h, yielding a 50 wt% solution of sodium pyrrolidone carboxylate. A 50 wt% solution of sodium pyrrolidone carboxylate (100.0 g, 0.33 mol, pH 7.7, optical purity 84%, L / D ratio = 92 / 8) was adjusted to pH 5.2 by adding nitric acid (purity 60 wt%, 2.7 g). An aqueous solution of zinc sulfate heptahydrate (47.6 g, 0.17 mol) in water (34.2 g) was added to the sodium pyrrolidone carboxylate solution (pH 4.1).The solution was mixed for 30 min at room temperature (pH 3.7), yielding crystals which were obtained by filtration. The crystals were washed with water (21.9 g) to give zinc pyrrolidone carboxylate dihydrate (32.0 g, 0.09 mol, yield 55%). The optical purity was 99.8% (L / D ratio 99.9 / 0.1). Preparation examples 1 to 40

[0061] The following are preparation examples using various preparations. These preparations were made using a conventional method. The amount to be added is given in wt.%.

[0062] The following are examples of how to prepare an ointment. Table 3 Preparation example 1 (ointment) Preparation example 2 (ointment) Preparation example 21 (ointment) Preparation example 22 (ointment) L-PCA zinc salt 0,001% - 0,001% 0,002% Zinc gluconate - 0,002% - - Ubiquinone - - 0,03% 0,02% Benzalkonium chloride 0,1% 0,1% 0,1% 0,1% urea 20,0% 20,0% 20,0% 20,0% White petroleum jelly 15,0% 15,0% 15,0% 15,0% Thin liquid paraffin 6,0% 6,0% 6,0% 6,0% Cetyl 3,0% 3,0% 3,0% 3,0% Stearyl alcohol 3,0% 3,0% 3,0% 3,0% Glyceryl monostearate 5,0% 5,0% 5,0% 5,0% aroma eq eq eq eq preservative eq eq eq eq buffer 1,0% 1,0% 1,0% 1,0% Purified water rest rest rest rest

[0063] The following are examples of how to prepare skin toners. Table 4 Preparation example 3 (skin toner) Preparation example 23 (skin toner) L-PCA zinc salt 0,03% 0,03% Ubiquinone - 0,03% Glycolic acid 5,0% 5,0% Glycerin 3,0% 3,0% Sorbit 2,0% 2,0% Polyoxyethylene(20)oleyl ether 1,0% 1,0% Ethanol 15,0% 15,0% Zinc paraphenolsulfonate 0,2% 0,2% buffer 0,1% 0,1% aroma 0,2% 0,2% preservative eq eq Purified water rest rest Table 5 Preparation example 4 (skin toner) Preparation example 5 (skin toner) Preparation example 24 (skin toner) Preparation example 25 (skin toner) DL-PCA zinc salt 0,005% - 0,005% - L-PCA zinc salt - - - 0,005% Zinc glycinate - 0,005% - - Ubiquinone - - 0,03% 0,01% citric acid 1,0% 1,0% 1,0% 1,0% urea 4,0% 4,0% 4,0% 4,0% Salicylic acid 2,0% 2,0% 2,0% 2,0% Lactic acid 2,0% 2,0% 2,0% 2,0% Glycerin 2,0% 2,0% 2,0% 2,0% Betaine 2,0% 2,0% 2,0% 2,0% Hyaluronic acid 0,1% 0,1% 0,1% 0,1% Ethanol 15,0% 15,0% 15,0% 15,0% buffer 0,1% 0,1% 0,1% 0,1% aroma 0,2% 0,2% 0,2% 0,2% preservative eq eq eq eq Purified water rest rest rest rest

[0064] The following are examples of how to prepare a lotion. Table 6 Preparation example 6 (lotion) Preparation example 7 (lotion) Preparation example 26 (lotion) Preparation example 27 (lotion) L-PCA zinc salt 0,01% - 0,01% 0,005% Zinc chloride salt - 0,005% - - Ubiquinone - - 0,03% 0,01% Lactic acid 0,1% 0,1% 0,1% 0,1% fruit acid 0,1% 0,1% 0,1% 0,1% Glycerin 4,0% 4,0% 4,0% 4,0% kaolin 1,0% 1,0% 1,0% 1,0% Calamin 0,7% 0,7% 0,7% 0,7% Camphor 0,2% 0,2% 0,2% 0,2% Ethanol 14,0% 14,0% 14,0% 14,0% aroma eq eq eq eq Purified water rest rest rest rest

[0065] The following are examples of how to prepare a cream. Table 7 Preparation example 8 (cream) Preparation example 9 (cream) Preparation example 28 (cream) Preparation example 29 (cream) L-PCA zinc salt 0,01% - 0,01% 0,02% Zinc gluconate - 0,02% - - Ubiquinone - - 0,005% 0,02% Resorcin 0,1% 0,1% 0,1% 0,1% Kojic acid 1,0% 1,0% 1,0% 1,0% Stearic acid 2,0% 2,0% 2,0% 2,0% Polyoxyethylene(25) cetyl ether 3,0% 3,0% 3,0% 3,0% Glyceryl monostearate 2,0% 2,0% 2,0% 2,0% Octyldodecanol 10,0% 10,0% 10,0% 10,0% Cetyl 6,0% 6,0% 6,0% 6,0% reduced lanolin 4,0% 4,0% 4,0% 4,0% Squalane 9,0% 9,0% 9,0% 9,0% 1,3-Butylene glycol 6,0% 6,0% 6,0% 6,0% Polyethylene glycol (1500) 4,0% 4,0% 4,0% 4,0% preservative eq eq eq eq aroma eq eq eq eq Purified water rest rest rest rest Table 8 Preparation example 10 (cream) Preparation example 30 (cream) DL-PCA zinc salt 0,01% 0,01% Ubiquinone - 0,01% Glycolic acid 2,0% 2,0% solid paraffin 5,0% 5,0% beeswax 10,0% 10,0% vaseline 15,0% 15,0% liquid paraffin 41,0% 41,0% 1,3-Butylene glycol 4,0% 4,0% Glyceryl monostearate 2,0% 2,0% Monolauric acid sorbitan polyoxyethylene(20) 2,0% 2,0% borax 0,2% 0,2% preservative eq eq aroma eq eq antioxidant eq eq Purified water rest rest

[0066] The following are examples of how to prepare an emulsion. Table 9 Preparation example 11 (emulsion) Preparation example 12 (emulsion) Preparation example 31 (emulsion) Preparation example 32 (emulsion) L-PCA zinc salt 0,02% - 0,02% 0,01% Zinc sulfate - 0,01% - - Ubiquinone - - 0,01% 0,02% Lactic acid 2,0% 2,0% 2,0% 2,0% Stearyl alcohol 0,5% 0,5% 0,5% 0,5% hydrogenated palm oil 3,0% 3,0% 3,0% 3,0% liquid paraffin 35,0% 35,0% 35,0% 35,0% Dipropylene glycol 6,0% 6,0% 6,0% 6,0% Polyethylene glycol (400) 4,0% 4,0% 4,0% 4,0% Sorbitans sesquioleate 1,6% 1,6% 1,6% 1,6% Polyoxyethylene(20) oleyl ether 2,4% 2,4% 2,4% 2,4% Carboxyvinylpolymer 1,5% 1,5% 1,5% 1,5% Potassium hydroxide 0,1% 0,1% 0,1% 0,1% Chelating agents eq eq eq eq preservative eq eq eq eq aroma eq eq eq eq Purified water rest rest rest rest

[0067] The following are examples of serum preparation. Table 10 Preparation example 13 (Serum) Preparation example 14 (Serum) Preparation example 33 (Serum) Preparation example 34 (Serum) L-PCA zinc salt 0,05% - 0,05% 0,005% zinc lactate - 0,005% - - Ubiquinone - - 0,05% 0,03% fruit acid 0,5% 0,5% 0,5% 0,5% Dipropylene glycol 5,0% 5,0% 5,0% 5,0% Polyethylene glycol (400) 5,0% 5,0% 5,0% 5,0% Ethanol 10,0% 10,0% 10,0% 10,0% Carboxyvinylpolymer 0,5% 0,5% 0,5% 0,5% sodium alginate 0,5% 0,5% 0,5% 0,5% Potassium hydroxide 0,2% 0,2% 0,2% 0,2% Monostearic acid polyoxyethylene(20)sorbitan 1,0% 1,0% 1,0% 1,0% Monooleic acid sorbitol 0,5% 0,5% 0,5% 0,5% Oleyl alcohol 0,5% 0,5% 0,5% 0,5% Placental extract 0,2% 0,2% 0,2% 0,2% Acetic acid dl-α-tocopherol 0,2% 0,2% 0,2% 0,2% aroma eq eq eq eq preservative eq eq eq eq Antibleach eq eq eq eq Purified water rest rest rest rest

[0068] The following are examples of how to prepare a face mask. Table 11 Preparation example 15 (face mask) Preparation example 16 (face mask) Preparation example 35 (face mask) Preparation example 36 (face mask) DL-PCA zinc salt 0,03% - 0,03% - Zinc gluconate - 0,03% - - L-PCA zinc salt - - - 0,03% Ubiquinone - - 0,03% 0,01% Isopropanol 2,0% 2,0% 2,0% 2,0% Polyvinyl alcohol 15,0% 15,0% 15,0% 15,0% Carboxymethylcellulose 5,0% 5,0% 5,0% 5,0% 1,3-Butylene glycol 5,0% 5,0% 5,0% 5,0% Ethanol 12,0% 12,0% 12,0% 12,0% Polyoxyethylene(20) oleyl ether 0,5% 0,5% 0,5% 0,5% aroma eq eq eq eq preservative eq eq eq eq buffer eq eq eq eq Purified water rest rest rest rest

[0069] The following are examples of how to prepare foundation. Table 12 Preparation example 17 (Foundation) Preparation example 37 (Foundation) DL-PCA zinc salt 0,1% 0,001% Ubiquinone - 0,03% Salicylic acid 0,5% 0,5% liquid paraffin 10,0% 10,0% Monooleic acid polyoxyethylene(20)-sorbitan 3,5% 3,5% Propylene glycol 3,0% 3,0% titanium oxide 9,0% 9,0% kaolin 24,0% 24,0% Talk 42,0% 42,0% Color pigment 3,0% 3,0% aroma eq eq preservative eq eq antioxidant eq eq

[0070] The following are examples of how to prepare liquid hand soap. Table 13 Preparation example 18 (liquid hand soap) Preparation example 38 (liquid hand soap) L-PCA zinc salt 0,1% 0,01% Ubiquinone - 0,5% Sodium lauryl sulfate 30,0% 30,0% Betaine 3,0% 3,0% Glycerol fatty acid esters 1,0% 1,0% Phenoxyethanol 1,0% 1,0% EDTA 0,1% 0,1% Purified water rest rest

[0071] The following are examples of how to prepare shampoo. Table 14 Preparation example 19 (shampoo) Preparation example 39 (shampoo) DL-PCA zinc salt 0,1% 0,1% Ubiquinone - 0,5% Polyoxyethylene(3)lauryl ether-sulfuric acid-triethanolamine 3,0% 3,0% Polyoxyethylene(3)lauryl ether sodium sulfate 6,0% 6,0% Sodium lauryl sulfate 1,5% 1,5% Lauric acid diethanolamide 3,0% 3,0% Lauryldimethylaminoacetic acid betaine 2,5% 2,5% cationic cellulose 0,2% 0,2% Distearic acid ethylene glycol 2,0% 2,0% aroma eq eq preservative eq eq Chelating agents eq eq buffer eq eq Purified water rest rest

[0072] The following are examples of how to prepare a hair lotion. Table 15 Preparation example 20 (hair lotion) Preparation example 40 (hair lotion) DL-PCA zinc salt 0,05% 0,05% Ubiquinone - 0,05% Lactic acid 0,02% 0,02% Oleyl alcohol 0,2% 0,2% liquid paraffin 0,5% 0,5% Ethanol 5,0% 5,0% Sorbit 4,0% 4,0% Polyoxyethylene(20)lauryl ether 2,5% 2,5% Monolauric acid sorbitan 0,5% 0,5% pigment 0,1% 0,1% preservative 0,1% 0,1% aroma 0,1% 0,1% Purified water rest rest Commercial applicability

[0073] The present invention can be used as a collagen synthesis promoter that is highly effective in the field of cosmetics. It has a collagen synthesis-promoting effect at a level equal to or greater than that of conventionally known retinoids, retinol, and retinoic acid, is free of teratogenicity and side effects, and is a safe collagen synthesis promoter. The following is a sequence protocol as an electronic document. This can be accessed in both DEPATISnet and the DPMA register.

Claims

[1] Use of zinc pyrrolidone carboxylate as a collagen synthesis promoter. [2] Use of zinc pyrrolidone carboxylate and ubiquinone as collagen synthesis promoters. [3] Use according to claim 2, wherein ubiquinone and zinc pyrrolidone carboxylate are contained in a weight ratio of 50:1 to 0.1:

1. [4] Use of a composition to promote collagen synthesis, comprising zinc pyrrolidone carboxylate and ubiquinone as active ingredients. [5] Use according to claim 4, comprising zinc pyrrolidone carboxylate in a concentration of 1 µM to 100 mM in the composition. [6] Use of a composition to promote collagen synthesis comprising 0.0001 to 10 wt% ubiquinone and 0.000035 to 3.5 wt% zinc pyrrolidone carboxylate. [7] Use of a composition according to one or more of claims 4 to 6, further comprising an N-acyl amino acid ester or an amino acid derivative. [8] Use of a composition according to one or more of claims 4 to 7, wherein the composition is an ointment, skin toner, lotion, cream, emulsion, serum, face mask, foundation, liquid hand soap, shampoo or hair lotion. [9] Use of a composition according to one or more of claims 4 to 8, wherein collagen is synthesized via the enhanced expression of an ascorbic acid transporter in human fibroblasts.

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