CN109731001B

CN109731001B - New application of new agaro-oligosaccharide and composition containing new agaro-oligosaccharide

Info

Publication number: CN109731001B
Application number: CN201910112630.0A
Authority: CN
Inventors: 张春毅; 曾润颖; 侯艳平; 王斐; 曾翰庭
Original assignee: Lannao Technology Xiamen Co ltd
Current assignee: Lannao Technology Xiamen Co ltd
Priority date: 2019-02-13
Filing date: 2019-02-13
Publication date: 2021-02-26
Anticipated expiration: 2039-02-13
Also published as: CN109731001A

Abstract

The invention relates to a novel application of novel agaro-oligosaccharide as a hair restorer or a hair conditioner and a composition containing the novel agaro-oligosaccharide, in particular to the application of the novel agaro-oligosaccharide in preparing products for improving and/or promoting hair growth or eliminating and/or relieving symptoms or conditions related to poor hair growth conditions. The new agaro-oligosaccharide has the advantages of clear mechanism, clear effect, no side effect, high safety and obvious effect. The composition containing the new agaro-oligosaccharide provided by the invention has remarkable anti-hair loss and hair growth effects, and has no irritation, no side effect and high safety.

Description

New application of new agaro-oligosaccharide and composition containing new agaro-oligosaccharide

Technical Field

The present invention relates to a new use of new agaro-oligosaccharide, in particular, it relates to a new use of new agaro-oligosaccharide as active component for improving hair growth, more specifically, it relates to a new use for preventing alopecia and promoting hair growth.

Background

Alopecia and baldness caused by the same are common phenomena, and mental stress caused by the same brings about a lot of troubles and obstacles to human relationships and social life. With the acceleration of pace of life, the increase of living pressure and environmental pressure, people suffering from alopecia are younger and younger at present, and the alopecia of women is gradually increased not only for men, so that a plurality of treatment medicines, hair care external products and hair growth external products for preventing alopecia are provided.

The reasons for alopecia are complex and are the combined action of various factors. The most common type of hair loss is "androgenic alopecia", otherwise known as "seborrheic alopecia", the main cause of which is that sebum secretion is regulated by androgens, and when sebum is abnormally secreted or abnormally produced, hair health is affected, thus causing hair loss. At present, the problems of scalp grease are generally solved by methods such as androgen control or cleaning in the medicines and hair care products. For example, taking a bautamide tablet (i.e., N- (1, 1-dimethylethyl) -3-oxo-4-aza-5 α -st-1-en-17 β -amide) inhibits androgen levels, promotes hair growth, and treats androgenic alopecia and baldness. Topical minoxidil (6- (1-piperidinyl) -2, 4-pyrimidinediamine, 3-oxide) is used to treat androgenic alopecia. The two medicines have obvious treatment effect, but simultaneously, the level of androgen is reduced, so that corresponding side effects such as erectile dysfunction, hyposexuality, rash, inflammation, pruritus and the like on external application are also generated.

Some cosmetics achieve hair loss prevention or hair growth promotion effects by adding plant extracts. For example, chinese patent document CN105640803A discloses an anti-dandruff, anti-alopecia and hair care product and a preparation method thereof, which utilizes plant essential oil to disinfect and resist bacteria and balance the oil secretion on the surface layer of skin; chinese patent document CN101810553A discloses a hair loss preventing and hair growth promoting composition, which comprises various plant extracts such as angelica sinensis, polygonum multiflorum, radix sileris, radix sophorae flavescentis, rhizoma zingiberis, notopterygium root and the like. According to the theory of traditional Chinese medicine, the extracts generally have the efficacies of dispelling wind, removing dampness, promoting blood circulation, reducing pathogenic fire and the like, but the functional components of the extracts are unclear, so the action principle is unclear, and the extracts are easy to cause adverse reactions such as allergy, irritation, dermatitis and the like due to the complex components. Therefore, the development of a new product having the effects of protecting hair, preventing hair loss, and promoting hair growth is still expected by those skilled in the art.

Seaweed extracts have a variety of functions, polysaccharides being key active ingredients. The seaweed oligosaccharide further prepared from the polysaccharide by the enzymolysis method has the advantages of small molecular weight, good solubility and various physiological activities. For example, new agaro-oligosaccharides prepared from red algae have been found to have physiological activities in various fields, including moisturizing, whitening, etc. in skin care products; regulating crop growth, preserving, etc. in agricultural application; the health food has effects of resisting oxidation, protecting liver, reducing blood sugar, reducing blood lipid, etc. The new agaro-oligosaccharide is extracted from red algae, is distinguished according to the degree of polymerization, and mainly comprises new agarobiose, new agarotetraose, new agarohexaose, new agarooctaose, new agarodecose, new agarododecaose and the like. However, the functions of the new agaro-oligosaccharide which is a single component or a mixed component or the oligosaccharide from other seaweeds for protecting hair and preventing and growing hair are not reported.

Disclosure of Invention

The purpose of the present invention is to provide a novel use of novel agaro-oligosaccharides as an active ingredient for improving hair growth, particularly for preventing hair loss and promoting hair growth. The present inventors have surprisingly found that the use of novel agaro-oligosaccharides or compositions comprising novel agaro-oligosaccharides is effective in improving the hair growth state, and in particular, in preventing hair loss and promoting hair growth.

To this end, in a first aspect, the present invention provides the use of a novel agaro-oligosaccharide as an active ingredient in the manufacture of a product for improving and/or promoting hair growth, or eliminating and/or alleviating a condition or condition associated with a poor hair growth condition.

Further, the new agaro-oligosaccharide comprises one or more of new agarobiose, new agarotetraose, new agarohexaose, new agarooctaose, new agarodecose and new agarododecaose. In one embodiment, the neoagaro-oligosaccharide is neoagarobiose, or neoagarotetraose, or neoagarohexaose, or neoagarooctaose, or neoagarodecose, or neoagarotetraose and neoagarohexaose, or neoagarooctaose, and neoagarooctaose.

Further, the condition or state associated with poor hair growth state refers to alopecia or slow hair growth rate.

Further, the alopecia refers to androgenetic alopecia.

Further, the product is a topical application product.

Further, the product is a topical application product for external application to the skin.

Further, the product is a pharmaceutical or cosmetic.

Further, the product is in the form of a solution, a cream, a paste, a cream, or a gel.

Furthermore, the new agaro-oligosaccharide accounts for 0.01 to 1 weight percent of the total weight of the product, preferably 0.01 to 0.5 weight percent, and more preferably 0.1 to 0.5 weight percent.

Further, the product comprises: (1) neoagaro-oligosaccharide, and (2) physiologically acceptable adjuvants.

Further, the physiologically acceptable auxiliary materials comprise small molecule polyol and nonionic surfactant.

Further, the small molecular polyol is one or more of glycerol, propylene glycol, butanediol and pentanediol, and is preferably glycerol.

Further, the nonionic surfactant is one or more of polyethylene glycol (PEG), tween and SPAN, preferably PEG, and further preferably PEG-400.

In a second aspect, the present invention provides a composition, characterized in that it comprises: (1) a novel agaro-oligosaccharide, and (2) a pharmaceutically or cosmetically acceptable excipient, wherein the novel agaro-oligosaccharide is present in the composition at a concentration that promotes hair growth, improves hair condition, and the composition is a pharmaceutical or cosmetic.

Further, the new agaro-oligosaccharide accounts for 0.01 to 1 weight percent of the total weight of the composition, preferably 0.01 to 0.5 weight percent, and more preferably 0.1 to 0.5 weight percent.

Further, the pharmaceutically or cosmetically acceptable auxiliary materials include small molecule polyols and nonionic surfactants.

Further, the nonionic surfactant is one or more of PEG, Tween and SPAN, preferably PEG, and further preferably PEG-400.

In a preferred embodiment, the glycerol represents from 1 to 6% by weight, preferably from 1 to 4% by weight, of the total weight of the composition; the PEG-400 is present in an amount of 0.5 to 5 wt%, preferably 0.5 to 2 wt%, based on the total weight of the composition.

In a specific embodiment, the composition consists of neoagarobiose, glycerol, PEG-400, and water.

Further, the composition is in the form of a solution, a cream, a paste, a cream, or a gel.

In a third aspect, the present invention provides the use of the novel agaro-oligosaccharides as a hair restorer or conditioner.

Further, the novel agaro-oligosaccharide is used as an active ingredient for protecting hair, preventing alopecia, and/or promoting hair growth.

Furthermore, the new agaro-oligosaccharide is used as an active component for improving the activity of the hair papilla cells; in particular, neoagaro-oligosaccharides are used to increase the viability of damaged hair papilla cells caused by androgens (such as dihydrotestosterone).

Further, the novel agaro-oligosaccharide is used for promoting the capability of hair papilla cells to secrete cytokines.

Further, the new agaro-oligosaccharide is used for promoting the growth of hair length and the increase of hair weight.

Neoagarobiose, neoagarotetraose, neoagarohexaose, neoagarooctaose, neoagarodecose and neoagarodecobiose belong to the class of neoagaro-oligosaccharides, are oligosaccharides extracted from red algae seaweed, can be prepared from red algae polysaccharide by an enzymatic hydrolysis method, and can be obtained commercially at present. The new agaro-oligosaccharide can be used in the fields of food, medicines, cosmetics and the like, and has the functions of moisturizing, resisting oxidation and the like when being used in cosmetics, but the functions of protecting hair, preventing hair loss and growing hair of a single component or a mixed component of the new agaro-oligosaccharide are not reported before the application date of the invention. The invention respectively treats new agaro-oligosaccharides with single components: neoagarobiose, neoagarotetraose, neoagarohexaose, neoagarooctaose, neoagarodecose, and neoagaro-oligosaccharides containing multiple components: experiments are respectively carried out on neoagarotetraose, neoagarohexaose, neoagarooctaose, neoagarodecose and neoagarodecose, and the cell model verifies that the neoagaro-oligosaccharide has the functions of preventing hair loss and growing hair.

When the neoagaro-oligosaccharide is further applied to an animal model, aiming at the skin environment (skin contains grease), two components of small-molecular polyol and surfactant are selected and compounded with neoagarobiose, and the penetration and dispersion of the neoagarobiose on natural skin are promoted by utilizing the compatibility of alcohol to water and oil and the dispersion characteristic of the surfactant, so that the neoagarobiose can effectively play a role when being applied to the natural skin.

The small molecular polyol used in the invention is a polyol with a carbon chain length of 10 or less. The small molecular polyol used for compounding can be selected from small molecular polyols which have the compatible characteristics of water and oil and the moisturizing property and are commonly used in medicines or cosmetics, and the small molecular polyols comprise glycerol, propylene glycol, butanediol, pentanediol and the like; surfactants may include polyethylene glycol, tween, SPAN, and the like. Glycerol and polyethylene glycol are preferred in the present invention due to a combination of safety, non-irritation, and approved use. The glycerol has the compatible characteristic of water and oil, is beneficial to the penetration of the new agaro-oligosaccharide, and simultaneously has the moisturizing effect and is beneficial to increasing the water required by the normal metabolism of the hair; third, glycerin is a well-recognized raw material for pharmaceuticals and cosmetics with high safety. Polyethylene glycol is a safe nonionic surfactant, is helpful for reducing surface tension, enhancing contact between the new agaro-oligosaccharide and glycerol on skin, promoting penetration of the new agaro-oligosaccharide and fully playing a role in promoting hair growth of the new agaro-oligosaccharide.

The auxiliary materials which can be used for compounding the new agaro-oligosaccharide are various, in order to provide a composition which is convenient to be further compounded with other auxiliary materials to form various products, the invention preferably selects a composition which is composed of the new agaro-disaccharide, the glycerol, the PEG-400 and the water and has the hair loss prevention and hair growth promotion effects, can be applied to the raw materials of hair care products, and can also be directly used as the hair care products.

In summary, compared with the prior art, the invention has the following advantages:

(1) the invention provides a new application of new agaro-oligosaccharide, and the new agaro-oligosaccharide has high safety, no side effect and no irritation when being used for preventing alopecia and growing hair. The existing products for preventing hair loss and nourishing hair mainly comprise finasteride, minoxidil and plant extracts, and the finasteride and minoxidil have strong side effects; plant extracts are prone to produce irritation due to their complex composition.

(2) The new agaro-oligosaccharide has clear mechanism and definite effect on the anti-hair loss and hair growth, and the new agaro-oligosaccharide is proved to be capable of relieving the inhibition of dihydrotestosterone on hair papilla cells and improving the activity of the hair papilla cells for the first time. The plant extract widely used in the prior art has complex components, some components even have bad or negative effects on hair loss prevention and hair care, the hair loss prevention and hair growth promotion effects are reduced, the extract effects in different batches are unstable, the cost is increased, and waste is caused.

(3) The invention provides a composition containing new agaro-oligosaccharide, which has the effects of protecting hair, preventing alopecia and promoting hair growth, and has the advantages of remarkable effect, no irritation, no side effect and high safety when the composition is used for preventing alopecia and growing hair.

Drawings

Various other advantages and benefits will become apparent to those of ordinary skill in the art upon reading the following detailed description of the preferred embodiments. The drawings are only for purposes of illustrating the preferred embodiments and are not to be construed as limiting the invention. Also, like reference numerals are used to refer to like parts throughout the drawings. In the drawings:

FIG. 1 is a graph of the effect of different concentrations of DHT on dermal papilla cell viability;

FIG. 2 shows the effect of new agaro-oligosaccharides with different polymerization degrees and different concentrations on the viability of hair papilla cells

FIG. 3 shows the effect of new agaro-oligosaccharides with different polymerization degrees on the expression level of VEGF when they act on an androgen alopecia in vitro cell model;

FIG. 4 shows the effect of DHT on the expression levels of DKK-1, TGF- β 2, β -catenin and AR when DHT acts on dermal papilla cells

FIG. 5 shows the effect of neoagarobiose (DP2) on the expression levels of DKK-1, TGF- β 2, β -catenin and AR when applied to an in vitro cell model of androgenetic alopecia;

FIG. 6 shows the effect of Neoagartetrakis and hexaose (DP4-6) on the expression levels of DKK-1, TGF- β 2, β -catenin and AR when they are applied to an in vitro cell model of androgenetic alopecia;

FIG. 7 shows the effect of Neoagarooctase, Neoagarodecose, and Neoagarodecabiose (DP8-12) on the expression levels of DKK-1, TGF- β 2, β -catenin, and AR in an in vitro cell model of androgenetic alopecia;

fig. 8 shows the results of transdermal permeation of neoagarobiose and compositions thereof.

Detailed Description

Exemplary embodiments of the present disclosure will be described in more detail below with reference to the accompanying drawings. While exemplary embodiments of the present disclosure are shown in the drawings, it should be understood that the present disclosure may be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.

The invention is described generally and/or specifically for the materials used in the tests and the test methods, in the following examples,% means wt%, i.e. percent by weight, unless otherwise specified. The reagents or instruments used are not indicated by manufacturers, and are all conventional reagent products which can be obtained commercially.

Example 1 Effect of Neoagaro-oligosaccharides on in vitro cell model of androgen alopecia

The method comprises the steps of taking hair papilla cells as a test object, stimulating and establishing an androgen alopecia in-vitro cell model by using Dihydrotestosterone (DHT) with a certain concentration, treating the androgen alopecia in-vitro model by using neoagaro-oligosaccharide, and evaluating the hair regeneration promoting effect of the neoagaro-oligosaccharide by detecting the content changes of Vascular Endothelial Growth Factor (VEGF), DKK-1, transforming growth factor (TGF-beta 2), beta-catenin (beta-catenin) and Androgen Receptor (AR) secreted by the hair papilla cells by using minoxidil as a positive control.

The new agaro-oligosaccharides used in this example had the following three types: (1) neoagarobiose (DP 2); (2) neoagarotetraose and neoagarohexaose (DP4-6), wherein the ratio of neoagarotetraose is 55% and the ratio of neoagarohexaose is 45%; (3) neoagarooctase, neoagarodecose and neoagarodecabiose (DP8-12), wherein the content of neoagarooctase is 45%, the content of neoagarocatadose is 30%, and the content of neoagarodecabiose is 25%

The specific test steps are as follows:

(I) construction of in vitro model of androgenetic alopecia

The hair papilla cells are taken as research objects, the hair papilla cells are treated by Dihydrotestosterone (DHT) with different concentrations, and the optimal stimulation concentration of the DHT is determined according to the change conditions of cell activity, cell factors VEGF, TGF-beta 2, beta-catenin, DKK1 and androgen receptor AR, wherein the optimal stimulation concentration of the DHT is a concentration which can enable the expression quantity of several secretory factors to change significantly (P is less than 0.01) on the premise of not damaging the cells (the cell activity is more than or equal to 90%). The method comprises the following specific steps:

(1) subjecting hair papilla cells to treatment at a ratio of 1.5X 10⁵The cells were seeded at a density of 96-well plates and placed at 37 ℃ in a 5% CO atmosphere₂Incubating in an incubator for 24 h;

(2) discarding the culture medium of cells cultured for 24h, replacing with new culture medium, adding DHT (0nM, 25nM, 50nM, 100nM, 200nM, 400nM, 600nM, 800nM, 1000nM) with different concentrations, and culturing for 24 h;

(3) and (3) cell viability detection: after the culture in step (2), the supernatant was discarded, 200. mu.L of MTT working solution with a concentration of 0.5mg/mL was added, and incubation was carried out at 37 ℃ in the absence of light. Discarding the supernatant after 4h, adding 150 μ L of dimethyl sulfoxide to each well, and reading OD with microplate reader₄₉₀The value is obtained. With blank control (0nM DHT added)The cell OD value is 100% of cell activity, the cell activity value is calculated, and the cell activity value after being treated by DHT with different concentrations is shown in figure 1.

As can be seen from FIG. 1, the viability of dermal papilla cells was maintained at 90% or more when the concentration of Dihydrotestosterone (DHT) was not higher than 800nM, and was reduced to 82.26% when the concentration of DHT reached 1000 nM. And the DHT concentration is 800nM, the expression level of the first five secreted factors can be changed significantly (P < 0.01), and the optimal DHT stimulation concentration is 800nM according to the above experimental results.

(II) detecting cytotoxicity of new agaro-oligosaccharides with different polymerization degrees on hair papilla cells

The toxic effect of the new agaro-oligosaccharides with different concentrations on the hair papilla cells is detected by an MTT method, and the safe working concentration range of the sample is screened out for carrying out subsequent efficacy evaluation experiments. The detection method comprises the following steps:

TABLE 1 table for concentration settings of new agaro-oligosaccharides

(1) Cell culture medium was added to 96-well plates at 1.5X 10⁵The cells were seeded at a density of 96-well plates and placed at 37 ℃ in a 5% CO atmosphere₂Incubating in an incubator for 24 h;

(2) preparing liquid: preparing a cell culture solution: experimental groups: according to the concentrations recorded in Table 1, the culture solution was used to prepare new agaro-oligosaccharides (DP2, DP4-6, DP8-12) with different degrees of polymerization to the desired concentrations; positive control group (PC group): preparing a culture solution containing 4% DMSO; blank group: the culture solution is not treated for standby;

(3) administration: discarding the cell culture solution cultured for 24h, respectively adding the experimental group culture solution, the PC group culture solution and the blank group culture solution prepared in the step (2), and continuously culturing for 24 h;

(4) and (3) detection: after 24h of cell incubation, discarding the supernatant, adding 200 μ L of MTT working solution with concentration of 0.5mg/mL, incubating at 37 ℃ in dark place, discarding the supernatant after 4h of incubation, adding 150 μ L of dimethyl sulfoxide to each well, and reading OD (optical density) by using a microplate reader₄₉₀The value is obtained. Calculating cell viability values of the experimental group and the PC group by taking the OD value of the cells of the blank control as 100% of cell viability, wherein the cell cytotoxicity detection results are shown in a table 2; the effect of different concentrations of neoagaro-oligosaccharides on the viability of hair papilla cells is shown in fig. 2.

TABLE 2 cytotoxicity assay results

From the cytotoxicity results (Table 2, FIG. 2), it was found that the new agaro-oligosaccharides with different degrees of polymerization had a comparable effect on the viability of hair papilla cells, and that the viability of hair papilla cells was maintained at 90% or more when the concentration thereof was within the range of 5000ppm, and was reduced to 90% or less when the concentration thereof was 10000 ppm. According to the results, the maximum safe administration concentration of the novel agaro-oligosaccharide on the hair papilla cells was 5000ppm, and it was finally determined that the hair papilla cells were administered by selecting three concentrations of 200ppm, 1000ppm and 5000ppm within the safe administration concentration range of the novel agaro-oligosaccharide.

(III) testing the effect of the new agaro-oligosaccharide on the in vitro cell model of androgen alopecia

Subjecting hair papilla cells to treatment at a ratio of 1.5X 10⁵The cells/well were seeded at a density in 6-well plates and placed at 37 ℃ with 5% CO₂Incubate for 24 h. The blank control group is not treated, 800nM DHT is added into the negative control group (NC) for stimulation, 500 mu M minoxidil and 800nM DHT are added into the positive control group (PC) at the same time, 800nM DHT and new agaro-oligosaccharides with different polymerization degrees are added into the experimental group at the same time, incubation is continued for 24h, supernatant and cells are collected respectively, and VEGF in the supernatant is detected by adopting an ELISA method; collecting cells, and detecting DKK-1, TGF-beta 2, beta-catenin and AR by RT-PCR. Graph Pad Prism was used for mapping and the results are expressed as Mean ± SD.

The detection result of VEGF is shown in figure 3, when the statistical analysis is carried out by using the T-Test method, the significance of the NC group is represented by # compared with the blank, the P-value < 0.05 is represented by #, and the P-value < 0.01 is represented by # #; significance was shown as P-value < 0.05 and P-value < 0.01 for each experimental group and PC group compared to NC group.

The results of DKK-1, TGF-beta 2, beta-catenin and AR detection are shown in FIGS. 4-7. By using 2^-△△CTResults were calculated by the method, and when statistical analysis was performed by the T-Test method, in FIG. 4, blank group mRNA amplification multiples were normalized, and compared with the NC group, significance was represented by #, # for P-value < 0.05, and # for P-value < 0.01. In fig. 5 to 7, the mRNA amplification fold of the NC group was normalized, and the significance of the PC group and each experimental group was shown as x, P-value < 0.05 and P-value < 0.01, respectively, compared to the NC group.

As can be seen from fig. 3, VEGF secretion was significantly reduced in the NC group compared to the blank group, indicating that DHT stimulation conditions were effective in this experiment. Compared with an NC group, the three concentrations of the neoagaro-oligosaccharide DP2 in the experimental group have obvious improvement effect on the secretion of VEGF, the improvement effect is more obvious along with the improvement of the concentration of the neoagaro-disaccharide, and when the concentration of the neoagaro-oligosaccharide DP2 reaches 5000ppm, the content of the VEGF even exceeds that of minoxidil; the new agaro-oligosaccharides DP4-6 and DP8-12 have no significant influence on the secretion amount of VEGF under the condition of low concentration (200ppm), and when the concentration of the two is increased, the secretion amount of VEGF is also significantly increased. This shows that the new agaro-oligosaccharides DP2, DP4-6 and DP8-12 can promote the secretion of VEGF in hair papilla cells, effectively relieve the inhibition of DHT on the cells, and DP2 has better performance than DP4-6 and DP 8-12.

As can be seen from FIG. 4, compared with the blank group, 800nM DHT has a significant inhibitory effect on the expression of beta-catenin gene in dermal papilla cells; the expression of TGF beta 2, DKK1 and AR genes of hair papilla cells is obviously improved, and the experiment proves that the DHT stimulation condition is effective.

From fig. 5, it can be seen that the new agaro-oligosaccharide DP2 can stimulate the expression of β -catenin gene in an androgen alopecia model and inhibit the expression of TGF- β 2, DKK1 and AR receptor, and the expression level is significantly different from that of the negative control group. As can be seen from FIG. 6, the new agaro-oligosaccharide DP4-6 has significant inhibitory effect on the expression of TGF-beta 2, DKK1 and AR receptors, but has no significant effect on the expression of beta-catenin gene. From FIG. 7, it can be seen that the novel agaro-oligosaccharide DP8-12 has significant inhibitory effect on the expression of DKK1 and AR receptors, but has no significant effect on the expression of beta-catenin and TGF-beta 2 genes.

By combining the analysis, the new agaro-oligosaccharides with three polymerization degrees have repairing effect on an androgen alopecia model caused by DHT, can improve the expression quantity of secretion factors related to alopecia in the model and show the effects of preventing alopecia and growing hair, wherein the new agaro-oligosaccharide DP2 has optimal performance, has significant influence on VEGF, beta-catenin, TGF-beta 2, DKK1 and AR, and has secondary effect (no significant influence on partial indexes) on the new agaro-oligosaccharides DP4-6 and DP 8-12. Therefore, neoagarobiose is preferably used as the main active ingredient of the hair loss preventing and growing composition.

EXAMPLE 2 transdermal penetration test

The percutaneous permeation test aims at investigating the permeability of the neoagarobiose on skin, and due to the barrier effect of cells, the neoagarobiose with high concentration is adopted to improve the content of the neoagarobiose in a sample after permeation, so that the detection accuracy is ensured. Meanwhile, glycerol and a non-ionic surfactant (such as polyethylene glycol) are added into the neoagarobiose to enhance the percutaneous absorption performance, and the selection and concentration of a penetration enhancer are designed as shown in the table 3.

TABLE 3 selection and concentration design of penetration enhancers

Combination 1	1% Neoagarobiose
		Combination
2	1% Neoagarobiose + 1% Glycerol
		Combination
3	1% Neoqiong disaccharide + 2% glycerol
		Combination
4	1% Neoagarobiose + 1% PEG400
		Combination 5	1% Neoagarobiose + 2% PEG400
Combination
	6	1% neoagarobiose, 1% glycerol and 1% PEG400
Combination 7			1% neoagarobiose, 2% glycerol and 1% PEG400

The treated SD rat skin was fixed in a diffusion cell of a transdermal diffusion apparatus (model RYJ-12B). 500 mu L of physiological saline containing 1% neoagarobiose is dripped on rat skin to avoid generating bubbles. A preservative film is added on the supply chamber to prevent the water from evaporating. 6mL of physiological saline is added into the receiving tank, the circulating water is kept at 37 (+ -0.5) DEG C, and the stirring speed is 300 rpm. And extracting all liquid from the receiving pool when the liquid permeates for 0.5, 1, 2, 3, 4 and 5 hours respectively, placing the liquid at 4 ℃ to be tested, simultaneously supplementing equivalent isothermal fresh physiological saline into the receiving pool, and setting three times of experiments. The percutaneous absorption capacity of the neoagarobiose containing different penetration enhancers was measured according to the above-mentioned procedures. And (3) concentrating all samples to be detected to 1mL by using a rotary evaporator, determining the content of the neoagarobiose in the samples by using a DNS method, and calculating the permeation quantity of a unit area. The results of transdermal permeation of neoagarobiose and compositions thereof are shown in fig. 8.

As can be seen from fig. 8, the permeation of neoagarobiose alone is slow, the permeation promoting effect is obvious when glycerin and a nonionic surfactant are added into the neoagarobiose, and the permeation promoting effect is better than that of glycerin or PEG400 alone when glycerin and PEG400 are simultaneously present, wherein the combination 7 (2% glycerin + 1% PEG400) shows the best permeation promoting effect, so that the neoagarobiose can rapidly and efficiently permeate through the skin of animals.

Example 3 Hair growth test in mice

18 guinea pigs were randomly divided into 3 groups (blank, test 1, test 2) of 6 animals each, wherein the blank was saline and the test 1 was neoagarobiose (containing 5000ppm of neoagarobiose)Sugar), test 2 group was composition group (5000ppm neoagarobiose + 2% glycerol + 1% PEG 400). Injecting appropriate amount of anesthetic into guinea pig, and after anesthesia, placing the same part of 3 × 3cm back of each guinea pig²The body hair is removed, the skin is wiped clean with physiological saline without damaging the skin, and the guinea pig is returned to the cage. After 24 hours, 500. mu.L of each hair removal part was applied uniformly 1 time per day for 30 consecutive days, 10 hairs were cut out from the administration site of guinea pigs at random 30 days after the administration, the length and weight of the hairs were measured, and the hair growth of each group of guinea pigs was compared with the average value thereof as an index, and the hair growth effect of mice was measured as shown in Table 4.

TABLE 4 Hair growth Effect of Neoagarobiose and compositions thereof on mice

As can be seen from the above table, neoagarobiose and its composition have a significant effect of promoting hair growth in depilated mice, compared to physiological saline. The hair growth effect of the composition containing the glycerol and the polyethylene glycol is more obvious, the hair growth rate reaches 53.5 percent in 30 days, and the hair weight is increased by 56.8 percent. This shows that substances such as glycerol and polyethylene glycol can effectively promote the diffusion of neoagarobiose in skin tissues, and the composition has a remarkable hair growth promoting effect.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.

Claims

1. Use of novel agaro-oligosaccharides as active ingredient for the preparation of a product for improving and/or promoting hair growth, or eliminating and/or alleviating hair loss or slow hair growth rate, said product being a product for topical application.

2. The use according to claim 1, characterized in that said neoagaro-oligosaccharides represent 0.01 to 1% by weight of the total weight of the product.

3. Use according to claim 1, characterized in that the product comprises physiologically acceptable adjuvants.

4. Use according to claim 3, characterized in that the physiologically acceptable adjuvants comprise small molecule polyols and non-ionic surfactants.

5. Use of neoagaro-oligosaccharides for the preparation of a hair tonic or hair conditioner wherein the neoagaro-oligosaccharides are used for increasing hair papilla cell viability, said hair tonic or hair conditioner being a product for topical application.

6. Use according to claim 5, wherein the neoagaro-oligosaccharides are used to increase the viability of damaged hair papilla cells caused by androgens.