TWI650128B - Matrix metalloproteinase inhibitor from symbiotic fermented product and application thereof - Google Patents

Abstract

The present invention relates to a matrix metalloproteinase inhibitor comprising a symbiotic yeast product, and the symbiotic yeast product is obtained by using a plurality of fermentation strains in a vegetative medium through a symbiotic fermentation step and a solid-liquid separation step. The obtained matrix metalloproteinase inhibitor can be applied to a skin external composition or to prepare an oral composition or a pharmaceutical composition which inhibits matrix metalloproteinase.

Description

Matrix metalloprotein derived from symbiotic yeast Enzyme inhibitors and their applications

The present invention relates to a matrix metalloproteinase inhibitor, and more particularly to a matrix metalloproteinase inhibitor derived from a symbiotic yeast product and its use in an oral composition, a skin external composition, and a pharmaceutical composition.

Matrix metalloproteinases are a general term for a group of matrix metallopoproteins (MMPs) that have the ability to cleave collagen and extracellular matrix (ECM). At least 28 different classes of matrix metalloproteinases have been discovered, each of which can decompose different extracellular matrices.

In addition to the degradation of extracellular matrix (ECM) and basement membrane, MMPs play an important role in the regulation of the synthesis and secretion of cytokines, growth factors, hormones and cell adhesion molecule receptors, and thus participate in development, evolution, and morphogenesis. , tissue remodeling, angiogenesis, arthritis, cardiovascular disease, stroke, multiple sclerosis, neurodegenerative diseases, allergies, and cancer A series of physiological and pathological processes. In the above MMP, type 2 MMP (MMP-2) belongs to gelatin enzyme, type 9 MMP (MMP-9) is particularly associated with tumor metastasis and angiogenesis, and MMP-2 and MMP-9 can promote microvascular regeneration and decomposition. Tissue, helping tumor cells to spread, migrate and infiltrate.

Past studies have shown that matrix metalloproteinases can be regulated by endogenous inhibitors such as matrix metalloproteinase inhibitors (TIMPs), alpha 2 macroglobulin heparin, and RECK proteins, thereby regulating the secretion of matrix metalloproteinases. However, the diversity of biological functions of endogenous inhibitors and the difficulty in systemic protein administration make endogenous inhibitors a major obstacle in clinical applications, so researchers try to screen from natural sources. A matrix metalloproteinase inhibitor is produced.

Among the products of natural origin, extracts such as Astragalus, Cinnamon, Euonymus, Magnolia, Uncaria, and sponge have the activity of inhibiting the expression of matrix metalloproteinases. However, the above-mentioned naturally-derived products having inhibitory matrix metalloproteinases are not highly selective for MMPs, have low oral availability, are unstable in metabolism, have high biotoxicity, have large side effects in clinical trials, and fail to achieve the desired effects.

In view of this, there is an urgent need to develop a product of natural origin to provide a product that inhibits the expression of matrix metalloproteinases.

Accordingly, one aspect of the present invention provides a matrix metalloproteinase inhibitor derived from a symbiotic yeast product comprising a symbiotic yeast product, and the symbiotic yeast product is a plurality of species in a vegetable medium.酦 The yeast strain is obtained by a symbiotic fermentation step and a solid-liquid separation step.

Another aspect of the present invention provides a skin external composition comprising the above-mentioned matrix metalloproteinase inhibitor as an active ingredient.

Still another aspect of the present invention provides a matrix metalloproteinase inhibitor for the preparation of an oral composition for inhibiting a matrix metalloproteinase, which comprises the above matrix metalloproteinase inhibitor as an active ingredient.

A further aspect of the present invention provides a matrix metalloproteinase inhibitor for preparing a pharmaceutical composition for inhibiting matrix metalloproteinases, wherein the matrix metalloproteinase inhibitor is used as an active ingredient, and the symbiotic fermentation product inhibits matrix metal The performance of proteases.

According to the above aspect of the present invention, a matrix metalloproteinase inhibitor derived from a symbiotic fermentation product is proposed, which is obtained from a phytochemical medium by a symbiotic fermentation step and a solid-liquid separation step in a vegetable medium. The symbiotic fermentation step is carried out at 37 ° C for 70 hours to 100 hours, and the aforementioned fermentation strain is composed of a plurality of lactic acid bacteria strains and at least one non-lactobacillus strain.

According to an embodiment of the present invention, the lactic acid bacteria strain comprises a plurality of plural Bifidobacterium spp. strains and/or Lactobacillus spp. strains.

According to an embodiment of the present invention, the above-described system of the genus Bifidobacterium bifurcated bifidobacteria (B.bifidum; BCRC 14615); and one of the group consisting of any combination thereof, Bifidobacterium longum (BCRC 14602 B.longum) .

According to an embodiment of the present invention, the Lactobacillus strain is a Lactobacillus delbrueckii subsp. bulgaricus (BCRC 10696), a Lactobacillus subfamily subspecies ( L. paracasei subsp. paracasei ; BCRC 17475) , a group consisting of Lactobacillus brevis ( L. brevis ; BCRC 12187), and Lactobacillus sp. (BCRC 17003) and any combination thereof.

According to an embodiment of the present invention, the at least one non-lactobacillus strain is composed of a strain of Streptococcus spp. and a strain of Saccharomyces spp .

According to an embodiment of the present invention, the Streptococcus sp. strain is Streptococcus salivarius subsp. thermophilus (BCRC 13869).

According to an embodiment of the present invention, the above-mentioned yeast strain is Saccharomyces cerevisiae (BCRC 21680).

According to an embodiment of the invention, the vegetable medium comprises soybean, sesame, alfalfa, sucrose, sodium chloride and water.

According to an embodiment of the present invention, the vegetable medium comprises 1.0 to 10.0% by weight of soybean, 1.0 to 5.0% by weight of sesame, 1.0 to 5.0% by weight of cerium, 1.0 to 5.0% by weight of sucrose, and 0.1 to 2.0% by weight. Sodium chloride.

According to another aspect of the present invention, a matrix metalloproteinase inhibitor is proposed for the preparation of an oral composition having a matrix metalloproteinase inhibitor, which comprises the above-mentioned matrix metalloproteinase inhibitor as an active ingredient.

Applying the matrix metalloproteinase inhibitor of the present invention, which comprises a symbiotic fermentation product, and the symbiotic fermentation product is planted The physical medium is obtained by a plurality of fermentation strains through a symbiotic fermentation step and a solid-liquid separation step, and the obtained matrix metalloproteinase inhibitor inhibits the expression of the matrix metalloproteinase, and can be used as an active ingredient and applied to the skin external composition, and the preparation is inhibited. An oral composition or pharmaceutical composition of a matrix metalloproteinase.

The above and other objects, features, advantages and embodiments of the present invention will become more <RTIgt; 1 and HPLC spectra of the symbiotic fermentation products obtained in Comparative Example 2.

2A to 2C are symbiotic yeast products showing the experimental examples (Fig. 2A), Comparative Example 1 (Fig. 2B), and Comparative Example 2 (Fig. 2C) of Example 1 having a concentration of 1.0 to 10% according to the present invention. Gelatin zymogram colloidal photograph of matrix metalloproteinases (MMP-2 and MMP-9) after co-culture with 3T3 fibroblasts.

As described above, the present invention provides a matrix metalloproteinase inhibitor derived from a symbiotic yeast product comprising a symbiotic yeast product, and the symbiotic yeast product is a plurality of fermentation strains used in a vegetable medium. It is obtained by a symbiotic fermentation step and a solid-liquid separation step.

In the present invention, the term "symbiotic fermentation product" as used herein refers to a product obtained by culturing various strains in vitro in a fermentation substrate for symbiotic fermentation, followed by solid-liquid separation, etc., wherein Contains residual components of cells, metabolites, etc.

In one embodiment, the aforementioned symbiotic fermentation product can be obtained, for example, in a vegetable culture medium, inoculated with a plurality of fermentation bacterium, and subjected to a symbiotic fermentation step and a solid-liquid separation step. It is explained here that not the symbiotic fermentation product naturally inhibits the expression of matrix metalloproteinase, which needs to be confirmed by experiments. Taking lactic acid bacteria fermentation products as an example, studies have shown that lactic acid bacteria fermentation products cannot inhibit the expression of type 2 matrix metalloproteinases and/or type 9 matrix metalloproteinases contained in cell culture fluids in vitro.

In one embodiment, the above vegetable medium may include, for example, soybean, sesame, alfalfa, sucrose, sodium chloride, and water. In one example, the above vegetable medium may comprise 1.0 to 10.0 weight percent soybean, 1.0 to 5.0 weight percent sesame, 1.0 to 5.0 weight percent bismuth, 1.0 to 5.0 weight percent sucrose, and 0.1 to 2.0 weight percent chlorine. Sodium, the rest is water. In another illustration, the above vegetable medium may comprise 1.0 to 6.0 weight percent soybean, 1.0 to 3.0 weight percent sesame, 1.0 to 3.0 weight percent bismuth, 2 to 5 weight percent sucrose, and 0.2 to 1.0 weight percent Sodium chloride, the balance is water.

Here, it is explained that if the amount of each component of the vegetable culture medium is outside the above range, it is not expected to obtain an symbiotic fermentation product which inhibits the expression of the matrix metalloproteinase without being confirmed by experiments.

In one embodiment, the fermentation strain is selected from at least 8 of a population consisting of a plurality of lactic acid bacteria strains and at least one non-lactobacillus strain. In an example, the inoculum amount of the above fermented strain in the vegetable medium may be, for example, 3 to 5 weight percent.

According to an embodiment of the present invention, the lactic acid bacteria strain is composed of a plurality of plural strains of Bifidobacterium spp. and Lactobacillus spp. strains.

In the above embodiment, the Bifidobacterium may be bifurcated by the bifidobacteria (B.bifidum; BCRC 14615) and Bifidobacterium longum e.g. (B.longum; BCRC 14602) as one of the group consisting of.

In the above embodiment, the Lactobacillus strains may for example be selected from the group consisting of Lactobacillus delbrueckii subsp. Bulgaricus (L.delbrueckii subsp bulgaricus;. BCRC 10696 , Lactobacillus paracasei subsp (L.paracasei subsp paracasei;. BCRC 17475 ), a group of Lactobacillus brevis ( L. brevis ; BCRC 12187) and Lactobacillus sp. (BCRC 17003).

In the above examples, at least one non-lactobacillus strain is composed of a strain of Streptococcus spp. and a strain of Saccharomyces spp .

In the above examples, the Streptococcus strain is Streptococcus salivarius subsp. thermophilus (BCRC 13869).

In the above examples, the Saccharomyces strain is Saccharomyces cerevisiae (BCRC 21680).

In one embodiment, the aforementioned symbiotic fermentation step can be carried out, for example, at 37 ° C for 70 hours to 100 hours, and preferably at 37 ° C for 72 hours to 96 hours to obtain a fermentation product. In another illustration, the solid-liquid separation step may be, for example, a filtration step or a centrifugation step to remove the solids of the fermented product to obtain a symbiotic fermentation product.

In other embodiments, between the symbiotic fermentation step and the solid-liquid separation step, the termination of the fermentation step is more selectively performed. In one example, the termination of the fermentation step can be carried out, for example, at 90 ° C for 15 minutes.

In one embodiment, the symbiotic fermentation product obtained above can be selectively subjected to conventional post-treatment steps, such as a concentration step, a decolorization step, and/or a microbial detection step, depending on actual needs.

The symbiotic yeast product obtained above is evaluated by an in vitro activity test and confirmed to inhibit the expression of matrix metalloproteinase, so that it can be further applied to compositions of various dosage forms. Suitable compositions can include, but are not limited to, oral compositions, topical compositions for skin, and/or pharmaceutical compositions. In one embodiment, the oral composition, the external composition for skin, and/or the pharmaceutical composition of the above-described symbiotic yeast product are symbiotic yeast products as an active ingredient, and the composition of the above dosage form inhibits matrix metalloproteinase which performed. It is stated herein that, in general, it is known that those having a matrix metalloproteinase expression in a naturally-derived product have a dose-and time-dependent correlation in the degree of inhibition of expression. However, the symbiotic yeast product obtained by the above-mentioned method has good water solubility as a matrix metalloproteinase inhibitor, has no dose-dependent cytotoxicity, and can completely inhibit matrix metal without dose and time dependence. Protease expression, thus providing a matrix metalloproteinase inhibitor with high biosafety, stability, bioabsorbability and effectiveness.

The following examples are used to illustrate the application of the present invention, and are not intended to limit the present invention. Those skilled in the art can make various changes without departing from the spirit and scope of the present invention. Retouching.

Example 1: Preparation of symbiotic fermentation products

1. Preparation of plant culture medium

First, the plant culture media of Examples and Comparative Examples 1 to 2 were prepared according to Table 1.

Prior to fermentation, the vegetable medium listed in Table 1 can be first sterilized by conventional autoclaving, and the sterilization process conditions can be carried out, for example, at 121 ° C for 40 minutes. Then, 8 different fermentation strains were inoculated separately into the plant medium of Table 1, wherein the total inoculum of the fermentation strain in the vegetable medium may be 3% (w/v) to 5% (w/v). .

In this embodiment, the bifurcated Po yeast strains include Bifidobacterium (B.bifidum; BCRC 14615), Bifidobacterium longum (B.longum; BCRC 14602), Lactobacillus delbrueckii subsp. Bulgaricus (L.delbrueckii subsp . bulgaricus; BCRC 10696), Lactobacillus paracasei subsp (L.paracasei subsp paracasei;. BCRC 17475 ), Lactobacillus brevis (L.brevis; BCRC 12187), Lactobacillus (Lactobacillus sp;. BCRC17003), thermophilic Streptococcus salivarius subsp. thermophilus (BCRC 13869) and Saccharomyces cerevisiae (BCRC 21680).

Next, the inoculated plant medium is subjected to a symbiotic fermentation step, which is carried out at 37 ° C for 72 hours to 96 hours. The resulting symbiotic yeast product obtains the resulting symbiotic fermentation product and performs subsequent evaluation after the termination of the fermentation step and the solid-liquid separation step.

Example 2: Evaluation of symbiotic fermentation products by high performance liquid chromatography

1. Analytical product extraction of symbiotic fermentation products

The symbiotic fermentation products obtained in the experimental examples of the first embodiment, the comparative examples 1 and the comparative example 2 were first extracted with 50% ethyl acetate, and then filtered with a plurality of layers (for example, 6 layers) of gauze, and baked at 45 ° C. After drying for 6 hours, the extracted product of the symbiotic fermentation product test sample was taken, and 1 mg of the sample was taken and dissolved in 2 mL of DMSO for high performance liquid chromatography (HPLC) analysis.

2. Analytical method for extracting products from symbiotic fermentation products

The HPLC analysis conditions are as follows, and the results are shown in Figure 1:

(i) Analysis column: Cosmosil C18

(ii) UV-VIS detector: Hitachi L-7420

(iii) Detection wavelength: 243 nm

(iv) Mobile phase: Eluent A: methanol/acetic acid (100/0.5)

(v) Eluent B: methanol/water/acetic acid (80/20/0.5)

(vi) Gradient elution conditions: the methanol concentration increases linearly from 80% to 84% in 0 to 15 minutes, then increases to 86% in 15 to 30 minutes, and increases to 88% in 30 to 40 minutes. It is further increased to 94% in 0 to 50 minutes and further increased to 100% in 50 to 70 minutes.

(vii) Flow rate: 0.7 mL/min.

Please refer to FIG. 1 , which is a HPLC chart of the symbiotic fermentation product obtained in the experimental example of the first embodiment, the comparative example 1 and the comparative example 2. It can be seen from the results of Fig. 1 that the peak pattern of the different plant culture media is different after being treated by the steps of symbiotic fermentation. The peak pattern of the symbiotic fermentation product of the experimental group 1 can be used as a reference and as a quality control standard for the production of symbiotic fermentation products in the future.

Example 3: Evaluation of the expression of matrix metalloproteinases by symbiotic fermentation products

Cell culture

First, mouse 3T3 fibroblasts (BCRC 60071) were cultured in Dulbecco's Modified Eagle's Medium containing 10% fetal calf serum (Hyclone) and 2.5% bovine calf serum (Hyclone). DMEM) medium and added 3.7% (w/v) sodium bicarbonate, 0.03% (w/v) glutamate (L-glutamine), 100 units of penicillin per ml and 100 mg of streptavidin Antibiotics such as streptomycin are cultured in a 10% CO ₂ incubator at 37 ° C, usually inoculated in a 75 cm ² culture dish at a cell density of 1 × 10 ⁶ cells, subcultured every three to four days and replaced with fresh Medium. After that, about the 10th generation of 3T3 cells were taken and cell experiments of symbiotic fermentation products at different concentrations were performed.

2. Matrix metalloproteinase expression inhibition assay (gelatin-based zymography)

3T3 mouse fibroblasts (BCRC60071) were co-cultured with the symbiotic yeast products of the experimental examples of Examples 1 and 1 and 2, or co-cultured with the aforementioned symbiotic fermentation products (control group). . After 1 or 2 days of culture (24 or 48 hours), the cell culture medium of each group is taken and concentrated to a concentration of about 100 times the original concentration using a commercially available concentrated centrifuge tube, such as Centricon 10 (Millipore), and the protein concentration is Bio. -Rad protein determination assay kit, about the same amount (10g) of protein, non-denatured in 10% Tris-glycine colloid (containing 0.1% collagen) (non-denaturing) protein electrophoresis analysis. After the electrophoresis was completed, the electrophoretic film was treated with 2.5% Triton X-100 for 30 minutes at room temperature to ease the rocking oscillation. The electrophoretic film was then subjected to developing buffer (Bio-Rad) at room temperature. The reaction was carried out for 30 minutes and then reacted at 37 ° C for at least 24 hours.

After the reaction was completed, the electrophoretic film was dyed with 0.1% Coomassie blue (dissolved in 9.2% acetic acid and 45.4% methanol) for 20 minutes at room temperature, then 9.2% acetic acid and 45.4% methanol. clear Wash for 10 minutes and repeat 2 times. After that, it is washed with 10% acetic acid and 10% methanol until the white bands representing the strong expression of matrix metalloproteinases appear, that is, the analysis step of the effect of the extract product of the symbiotic fermentation product on the performance of the matrix metalloproteinase is completed. The results are shown in Figures 2A to 2C.

2A to 2C, which show the symbiotic fermentation of the experimental example (Fig. 2A), the comparative example 1 (Fig. 2B), and the comparative example 2 (Fig. 2C) of Example 1 according to the present invention at a concentration of 1.0 to 10%. Gelatin zymogram colloidal photograph of matrix metalloproteinases (MMP-2 and MMP-9) after co-culture with 3T3 fibroblasts. In Figures 2A to 2C, the Mth lane represents the protein marker, and the C0, _C24 , and _C48 lanes show the MMP expression of the untreated 3T3 mouse fibroblasts after ₀ , ₂₄ , and ₄₈ hours of culture, respectively.

2A to 2C, the symbiotic fermentation product of the experimental example (Fig. 2A) of the first embodiment of the present invention is at a concentration of 2.5 to 10% (24, 48 hours) for the enzyme of matrix metalloproteinase (MMP). Both -2 and MMP-9) showed significant inhibition and even almost completely inhibited their performance. Here, FIG. 2A shows not only the symbiotic fermentation product of the experimental example of Example 1 in a low dose (2.5%) experimental example ( FIG. 2A ), but also within 24 hours. Completely inhibited the expression of MMP-2 and MMP-9, and significantly inhibited the expression of MMP-2 and MMP-9 at 2.5~10%, and the inhibitory effect was not dose-dependent between 2.5 and 10%. . The inhibitory effect of the symbiotic fermentation product of the experimental example of the present invention on MMP-2 and MMP-9 is apparently more effective than other matrix metalloproteinase inhibitors known to have a dose- and time-dependent natural source product. As for the symbiotic fermentation product of Comparative Example 1 (Fig. 2B) at a concentration of 10% (treatment) At 24 and 48 hours, the expression of matrix metalloproteinase enzymes (MMP-2 and MMP-9) was inhibited. The symbiotic fermentation product of Comparative Example 2 (Fig. 2C) was at a concentration of 2.5-10% (treatment 24, 48 hours) Inhibition of matrix metalloproteinase enzymes (MMP-2 and MMP-9), but the inhibitory effect was far less than the experimental example (Fig. 2A).

Since the symbiotic fermentation product of the experimental example of Example 1 can significantly inhibit the expression of MMP-2 and MMP-9, it has a better protective effect on collagen, that is, it has the effect of delaying the destruction of collagen by matrix metalloproteinase due to aging factors. The anti-aging ability can be used as an active ingredient and applied to a skin external composition such as a cosmetic composition.

Next, the symbiotic fermentation product of the experimental example of Example 1 was administered at a low dose (3.5 g/kg body weight/day) and a medium dose (7 g/ according to the health food safety assessment method announced by the Food and Drug Administration of the Ministry of Health and Welfare of Taiwan. The dose of kg body weight/day and high dose (11.67g/kg body weight/day) was administered orally to the test animals (male and female rats, at least three replicates) using a gastric tube for 28 days of subchronic toxicity. The trial did not cause death and adverse clinical signs. Daily weight gain and water, feed consumption, morphology, behavioral activity, and organ tissue weight did not produce significant dose-related changes. The results of blood analysis and biochemical index test showed that there was no significant difference between the symbiotic fermentation product of the experimental example of Feeding Example 1 and the comparative example or the control group, which confirmed that there was no obvious subchronic toxicity and met the food safety standard (not shown). The results show that the no-observed-adverse-effect-level (NOAEL) of the symbiotic fermentation product of the experimental example of Example 1 was 11.67 g/kg/day.

Since the symbiotic fermentation product of the experimental example of Example 1 can significantly inhibit the expression of MMP-2 and MMP-9 and meet the food safety standards, it can be used as an active ingredient and used for preparing an oral composition which inhibits the expression of matrix metalloproteinase. The drug and/or the pharmaceutical composition is even used for inhibiting the outward spread, migration and infiltration of tumor cells.

In summary, the above examples demonstrate that the present invention is derived from a matrix metalloproteinase inhibitor of symbiotic fermentation products, which is successfully obtained by a plurality of fermentation strains through a symbiotic fermentation step and a solid-liquid separation step, and the obtained The symbiotic fermentation product does inhibit the expression of matrix metalloproteinases, and further inhibits the outward diffusion, migration and infiltration of tumor cells, and can be applied to external compositions for skin, and to prepare oral compositions or pharmaceutical compositions having inhibitory matrix metalloproteinases.

It should be noted that the present invention is exemplified by a specific process, a specific analytical method or a specific instrument, and illustrates the symbiotic fermentation product of the present invention for inhibiting the expression of matrix metalloproteinase and its application, but any of the technical fields of the present invention. It is to be understood by those skilled in the art that the present invention is not limited thereto, and that the symbiotic fermentation product of the present invention which inhibits matrix metalloproteinase expression may be used in other processes, other analytical methods, or the like without departing from the spirit and scope of the present invention. The instrument is carried out.

It can be seen from the above examples that the matrix metalloproteinase inhibitor derived from the symbiotic yeast product of the present invention and the use thereof have the advantage that the matrix metalloproteinase inhibitor is a symbiotic fermentation step and a solid-liquid separation using a plurality of fermentation strains. In the step, the obtained symbiotic yeast product inhibits the expression of matrix metalloproteinase, and can be applied to the external composition of skin, and the preparation has inhibition An oral composition or pharmaceutical composition of a matrix metalloproteinase.

While the invention has been described above in terms of several embodiments, it is not intended to limit the scope of the invention, and the invention may be practiced in various embodiments without departing from the spirit and scope of the invention. The scope of protection of the present invention is defined by the scope of the appended claims.

Claims (11)

A matrix metalloproteinase inhibitor derived from a symbiotic yeast product, which is obtained by a phytochemical fermentation step and a solid-liquid separation step in a vegetative medium, wherein the symbiotic fermentation step The strain is carried out at 37 ° C for 70 hours to 100 hours, and the fermentation strains are composed of a plurality of strains of lactic acid bacteria and at least one strain of non-lactobacillus strains, which are composed of a plurality of strains of Bifidobacterium (. Bifidobacterium spp) (. Lactobacillus spp) and a plurality of kinds of strains of Lactobacillus strain composition, the plurality of lines by the genus Bifidobacterium bifurcated bifidobacteria (B.bifidum; BCRC 14615), Bifidobacterium longum (B. longum; BCRC 14602) consisting of, the plurality of lines of Lactobacillus delbrueckii subsp. bulgaricus (L.delbrueckii subsp bulgaricus;.. BCRC 10696), Lactobacillus paracasei subsp (L.paracasei subsp paracasei; BCRC 17475 ), Lactobacillus brevis (L.brevis; BCRC 12187) and Lactobacillus (Lactobacillus sp;. BCRC17003) composed of the at least one non-line by a strain of Lactobacillus Streptococcus (Streptococcus spp) and yeast strains. (. Saccharomyces spp) composed of strain of the genus, the Streptococcus strain is Streptococcus thermophilus (Streptococcus salivarius subsp thermophilus;. BCRC 13869), and the Saccharomyces cerevisiae strain (Saccharomyces cerevisiae; BCRC 21680), and The matrix metalloproteinase inhibitor inhibits the expression of type 2 matrix metalloproteinases and/or matrix metalloproteinases type 9. According to the scope of claim 1 A matrix metalloproteinase inhibitor of the bacterium fermentation product, wherein the fermentation bacterium is inoculated in an amount of from 3 to 5 weight percent of the plant medium. A matrix metalloproteinase inhibitor derived from a symbiotic yeast product according to claim 1, wherein the vegetable medium comprises soybean, sesame, alfalfa, sucrose, sodium chloride and water. A matrix metalloproteinase inhibitor derived from a symbiotic yeast product according to claim 3, wherein the vegetable medium comprises 1.0 to 10.0% by weight of the soybean, 1.0 to 5.0% by weight of the sesame, 1.0 to 5.0% by weight of the cerium, 1.0 to 5.0% by weight of the sucrose, and 0.1 to 2.0% by weight of the sodium chloride. A matrix metalloproteinase inhibitor derived from a symbiotic yeast product according to claim 3, wherein the vegetable medium comprises 1.0 to 6.0% by weight of the soybean, 1.0 to 3.0% by weight of the sesame, 1.0 to 3.0% by weight of the bismuth, 2 to 5% by weight of the sucrose, and 0.2 to 1.0% by weight of the sodium chloride. A matrix metalloproteinase inhibitor for use in the preparation of an oral composition having an inhibitory effect on a matrix metalloproteinase, which is an active ingredient of the matrix metalloproteinase inhibitor according to any one of claims 1 to 5, and An effective dose of the matrix metalloproteinase inhibitor to the topical composition of the skin is from 3.5 g/kg body weight/day to 11.67 g/kg body weight/day. Matrix metal according to item 6 of the patent application scope A protease inhibitor is used to prepare an oral composition having an inhibitory effect on a matrix metalloproteinase, wherein the matrix metalloproteinase inhibitor inhibits the expression of a matrix metalloproteinase type 2 and/or a matrix metalloproteinase type 9. A topical composition for skin, comprising the matrix metalloproteinase inhibitor according to any one of claims 1 to 5, wherein the matrix metalloproteinase inhibitor is an active ingredient, and the matrix metalloproteinase inhibitor is One of the oral compositions has an effective dose of 2.5 to 10%. The composition for external use on skin according to the invention of claim 8, wherein the matrix metalloproteinase inhibitor inhibits the expression of the type 2 matrix metalloproteinase and/or the matrix metalloproteinase type 9. A matrix metalloproteinase inhibitor for use in the preparation of a pharmaceutical composition having a matrix metalloproteinase inhibitor, which is an active ingredient of the matrix metalloproteinase inhibitor according to any one of claims 1 to 5. A matrix metalloproteinase inhibitor according to claim 10, for use in the preparation of a pharmaceutical composition having a matrix metalloproteinase inhibitor, wherein the matrix metalloproteinase inhibitor inhibits matrix metalloproteinase type 2 and/or matrix metal of type 9 The performance of proteases.