KR102534846B1 - Seaweed extract with antibacterial activity against mycobacterium tuberculosis - Google Patents

Abstract

The present invention relates to a method for producing a seaweed extract with antibacterial activity against Mycobacterium tuberculosis. According to the present invention, a seaweed extract is produced which is a dry powder extracted and concentrated from one or more seaweed powders, selected from a group of Colpomenia bullosa, Scytosiphon canaliculatus, Scytosiphon lomentaria, Sargassum horneri, Sargassum muticum, Sargassum thunbergii, Padina arborescens, Sargassum micracanthum, Myagropsis myagroides, Laurencia okamurae, Ulva intestinalis, Phyllospadix japonica, through a series of steps using 70 % ethanol or a distilled water extraction solvent. In addition, as the seaweed extract is designed to inhibit the growth of Mycobacterium tuberculosis, problems caused by conventional chemotherapy drugs, drug-resistant tuberculosis, or emergence of drug-resistant strains are significantly overcome.

Description

Seaweed extract having antibacterial activity against Mycobacterium tuberculosis

The present invention relates to a seaweed extract having antibacterial activity against Mycobacterium tuberculosis, and more particularly, by providing a seaweed extract having an antibacterial activity against Mycobacterium tuberculosis from seaweed collected from nature, thereby preventing chemotherapeutic agents, drug-resistant tuberculosis, or drugs. It has a significant effect of improving various problems according to the emergence of resistant strains.

In general, tuberculosis is a chronic infection caused by Mycobacterium tuberculosis, which occurs when the body's resistance is weakened. Tuberculosis is asymptomatic in about 1/3 of humanity, so the risk of developing tuberculosis is higher in patients with reduced immune function. It increases.

Mycobacterium tuberculosis can cause symptoms by invading most tissues or organs, such as the kidneys, nerves, and bones. Tuberculosis occurs mainly in the lung tissue because the tuberculosis bacillus discharged from the cough, runny nose, and phlegm of an infected person is easily transmitted through the air. It proliferates by using nutrients from its host, the human body, and symptoms such as coughing, expectoration, chills, cold sweats, and weight loss appear. Once the symptoms develop, it is a disease with a very high fatality rate if left untreated.

Tuberculosis is diagnosed with an X-ray test, and if diagnosed with tuberculosis, it is treated with anti-tuberculosis drugs. Anti-tuberculosis drugs are oral first-line anti-tuberculosis drugs that take isoniazid and rifampin for a long period of time (6 months to 2 years), and kanamycin, streptomycin, levofloxacin, prontionamide, cycloserine, and linezolid to prevent insufficient treatment or resistance. , clofazimine and other anti-tuberculosis drugs are used in combination.

Compositions for treatment of various types of treatment for tuberculosis are being studied and produced based thereon.

With reference to the conventionally known technology, the composition and action of the anti-tuberculosis composition are schematically as follows.

Korean Patent No. 10 - 1352806 discloses that it is used together with one or more chemotherapeutic agents selected from isoniazid and rifampin, which are effective for the treatment of Mycobacterium tuberculosis infection, in preventing or delaying the reactivation of tuberculosis in mammals with latent infection of Mycobacterium tuberculosis. A pharmaceutical composition comprising an Mtb72f fusion protein or an immunogenic fragment thereof from the species Mycobacterium in the Mycobacterium tuberculosis group, and an adjuvant, wherein said one or more chemotherapeutic agents are initially administered to a mammal over a predetermined period of time, Then, it provides a method for preventing or treating Mycobacterium tuberculosis infection in which the pharmaceutical composition is administered.

In addition, Korean Patent Registration No. 10 - 1118942 discloses a 2,3-dihydro-6-nitroimidazo[2,1-b]oxazole compound, an optically active form thereof, and a pharmacologically acceptable salt thereof in the group consisting of Provided is an antituberculosis composition containing at least one selected from the group consisting of rifampicin, rifabutin, rifapentine, isoniazid, ethambutol, pyrazinamide and salts thereof.

On the other hand, in another prior art, a technology related to an anti-tuberculosis composition using a natural substance as a raw material is known. For example, Korean Patent Registration No. 10 - 0656969 discloses Japanese pepper tree, coriander tree, oriental herb pepper, flowering plant, or cheoncho. A pharmaceutical composition for the treatment of tuberculosis comprising a crude extract as an active ingredient, a crude pepper extract extracted from water, lower alcohol or a mixed solvent thereof, and containing 0.1 to 80% by weight of the crude extract as an active ingredient. are starting

As another example, Korean Patent Publication No. 10 - 2013 - 0029209 discloses the step of preparing chrysanthemum, water, ethanol or a mixture of water and ethanol and normal hexane, ether, chloroform, methylene chloride, normal butanol and methanol in a container. Introducing with any one solvent among organic solvents, extracting chrysanthemum extract by raising the temperature of the container, filtering, concentrating, and drying the chrysanthemum extract into powder, dispersing the powder with purified water and degreasing to separate the solvent A method for preparing an anti-tuberculosis composition comprising the step is also disclosed.

As another anti-tuberculosis composition having antibacterial activity against Mycobacterium tuberculosis using a natural substance, Korean Patent No. 10-1765988 "Anti-tuberculosis composition for treatment and prevention of tuberculosis containing an extract or a fraction thereof" is known.

Korean Registered Patent No. 10 - 1352806 (2014.02.17) Korean Registered Patent No. 10 - 1118942 (2012.02.27) Korean Registered Patent No. 10 - 0656969 (2006.12.13) Korean Patent Publication No. 10 - 2013 - 0029209 (2013.03.22) Korean Registered Patent No. 10 - 1765988 (2017.08.01)

Conventionally, chemotherapeutic agents such as isoniazid and rifampin have been basically used for the treatment and improvement of tuberculosis.

However, according to a report recently published by the World Health Organization (WHO), the prevalence of anti-tuberculosis drug-resistant tuberculosis is increasing worldwide, raising serious concern. The need for the development of new anti-tuberculosis agents for tuberculosis is emerging.

Based on this point, it is considered as a variety of therapeutic extracts of natural origin, among which marine organisms are known as a rich source of polyunsaturated fatty acids (PUFAs), polysaccharides, natural pigments (NPs), enzymes and bioactive peptides. In particular, marine brown algae, a sulfated polysaccharide, have been reported to exhibit various biological activities such as antiviral, antifungal, immunomodulatory, antioxidant, antitumor, antiinflammatory, antiangiogenic, and anticoagulant, and are therefore very suitable for application to antituberculosis extracts. judged as

In addition, in the case of various types of marine organisms, the cost of procurement of the materials is low, mass production is possible, and generally inedible marine organisms can be used, so the economic feasibility is very high and the utilization of discarded marine resources is increased. can make it

Accordingly, the present invention provides a seaweed extract having an antibacterial activity against Mycobacterium tuberculosis from such seaweed to make meaningful efforts to overcome various side effects and resistance problems of conventional chemotherapeutic agents and to secure economic feasibility.

Accordingly, the present invention was invented to solve the problems of the prior art as described above,
Washing seaweed collected from the sea, freeze-drying, and pulverizing to obtain seaweed powder (S10);
Obtaining a primary seaweed extract as a supernatant excluding seaweed residues by adding 70% ethanol to the seaweed powder obtained in the step of obtaining the seaweed powder (S10), stirring, and centrifuging (S20);
Adding 70% ethanol to the seaweed residue from the step of obtaining the first seaweed extract (S20), stirring, and centrifuging to obtain a second seaweed extract as a supernatant (S30);
The first seaweed extract obtained in the step of obtaining the first seaweed extract (S20) and the second seaweed extract obtained in the step of obtaining the second seaweed extract (S30) were mixed and vacuumed to 70% of the weight of the mixture. Concentrating (S40);
Freeze-drying the seaweed extract concentrated in the vacuum concentration step (S40) to obtain a seaweed extract having a growth inhibitory action against Mycobacterium tuberculosis (S50);
In the step of obtaining the seaweed powder (S10), the seaweed is at least one selected from the group consisting of long bule end, smooth round head, constricted end round head, hoesaeng cap, and mongdan leaf cap, Preparation of a seaweed extract having antibacterial activity against Mycobacterium tuberculosis make up the way
In addition, washing seaweed collected from the sea, freeze-drying, and pulverizing to obtain seaweed powder (S110);
Obtaining a first seaweed extract by adding tertiary distilled water to the seaweed powder obtained in the step of obtaining the seaweed powder (S110) and heating at 90 to 100 ° C (S120);
Cooling the first seaweed extract obtained in the step of obtaining the first seaweed extract (S120) at room temperature, separating the seaweed residues with a sieve, and obtaining a second seaweed extract as a supernatant (S130);
vacuum-concentrating the secondary seaweed extract obtained in the step of obtaining the secondary seaweed extract (S130) to 50% by weight (S140);
Freeze-drying the seaweed extract concentrated in the vacuum concentration step (S140) to obtain a seaweed extract having a growth inhibitory action against Mycobacterium tuberculosis (S150);
In the step of obtaining the seaweed powder (S110), the seaweed has antibacterial activity against Mycobacterium tuberculosis, characterized in that it is at least one selected from the group consisting of streptococci, sphagnum, small thorns, solitary larvae, double-headed lice, intestinal greens, and crab seas. Eggplant constitutes a method for producing a seaweed extract.

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Therefore, the present invention can achieve the object of producing a seaweed extract useful for improving various problems caused by the emergence of conventional chemotherapeutic agents, drug-resistant tuberculosis, or drug-resistant strains through growth inhibitory action against Mycobacterium tuberculosis. .

The present invention provides a seaweed extract having an antibacterial activity against Mycobacterium tuberculosis from naturally collected seaweed.

In particular, the present invention is antiviral, antifungal, immunomodulatory, antioxidant, antitumor, There is an effect of providing an extract having an antibacterial activity against Mycobacterium tuberculosis by using natural seaweed having various biological activities such as anti-inflammatory.

In addition, the present invention minimizes problems such as various side effects of conventional chemotherapeutic agents, such as peripheral neuropathy, hepatotoxicity, skin rash, and headache, and as a natural agent, it is possible to relieve and improve tuberculosis symptoms without fear of resistance. There is an advantage.

1 is a graph showing the results of measuring the antibacterial activity of Mycobacterium tuberculosis by FU of a seaweed extract having antibacterial activity against Mycobacterium tuberculosis according to the present invention by ethanol extraction as the first method and hot water extraction as the second method.
Figure 2 is a flow chart showing a first method of extracting seaweed having antibacterial activity against Mycobacterium tuberculosis according to the present invention.
Figure 3 is a flow chart showing a second method of extracting seaweed having antibacterial activity against Mycobacterium tuberculosis according to the present invention.

Hereinafter, the configuration and action according to a preferred embodiment of the seaweed extract having antibacterial activity against Mycobacterium tuberculosis of the present invention will be described in detail with reference to the accompanying drawings.

1 is a graph showing the results of measuring the antibacterial activity of Mycobacterium tuberculosis by FU of a seaweed extract having antibacterial activity against Mycobacterium tuberculosis according to the present invention by ethanol extraction, which is the first method, and hot water extraction, which is the second method, according to the present invention; FIG. is a flow chart showing manufacturing steps of the first method of the present invention, and FIG. 3 is a flowchart showing manufacturing steps of the second method of the present invention, which are described sequentially and together.

In the following description, detailed descriptions of parts that can be easily implemented by those skilled in the art may be omitted.

In addition, since the following description describes the present invention with respect to preferred embodiments, the present invention is not limited by the following examples, and it is natural that various modifications may be provided without departing from the scope of the present invention. will be.

The basic idea of the invention is that the seaweed extract having antibacterial activity against Mycobacterium tuberculosis, to which the technology of the present invention is applied, relates to a seaweed extract having an antibacterial activity against Mycobacterium tuberculosis from a number of naturally collected species, in particular seaweeds defined below. .

Marine organisms are rich sources of polyunsaturated fatty acids (PUFAs), polysaccharides, natural pigments (NPs), enzymes and physiologically active peptides, and in particular, seaweeds are reported to have various biological activities.

Seaweeds are classified into brown algae, red algae, and green algae according to habitat and color.

Brown algae such as japonica, rhubarb, hijab, seaweed, and kelp contain ingredients such as alginic acid and sulfide polysaccharide (fucoidan), and are known to have antiviral, immunomodulatory, antioxidant, antitumor, antiinflammatory, antiangiogenic, and anticoagulant activities. there is. It is known that red algae such as agar-agar, carrageenan, and seaweed contain antibacterial, antifungal, and anticancer substances, and green algae such as green algae, cheongcheong, and cheongtae contain a large amount of beta-carotene, an antioxidant.

The seaweed extract having antibacterial activity against Mycobacterium tuberculosis of the present invention is provided as a concentrated dry powder after extracting powder from seaweed using an extraction solvent, thereby inhibiting the growth of Mycobacterium tuberculosis, a major Mycobacterium tuberculosis. configure to do

The seaweeds are rhubarb (Ulleungdo), rhubarb (Yeongdeok), seaweed, half cap, half worm, half thorn, half half thorn, half half thorn, half thorn, half thorn, half agar, dog worm, ring knot crab, soft sticky, double-headed iseosil . It is composed of at least one selected from the group consisting of flesh, wrinkled hair, small bead coral end, gaumu, slippery nuari, hole galae, spiny seaweed, and Sakai stem end.

Rhubarb (Eisenia bicyclis) is a seaweed of the brown algae kelp family, and was used in the present invention collected from Ulleungdo and Yeongdeok, Gyeongsangbuk-do.

Seaweed, half cap, half thorn cap, half single leaf cap, half leaf cap, half leaf cap, half cap of hoesaeng, half half leaf cap, earthworm, long bulge end, smooth ring, narrow ring, and hibiscus are also brown algae plants, containing a large amount of fucoidan, and rami It is a known fact that it is rich in ingredients such as narin, polyphenol, histamine, alginic acid, vitamin C, and iodine, and has antioxidant, anti-allergic, antiviral, immunity enhancing, anti-inflammatory, and antibacterial physiological activities.

Furthermore, black gab, gab gab, agar-agar, ring knot crab, yellow sticky, double-headed seosil, gae-seo-sil, cow hair red thread, red black flesh, wrinkled hair, small bead coral end, gaewoo radish, slippery red algae seaweed It is known to contain a large amount of carrageenan, and is rich in alginic acid, polysaccharide sulfate, laminin, agarose, porphyran, vitamin A and minerals, and has antiviral, antioxidant, and immune-enhancing physiological activities.

Green seaweed, seaweed of green algae, contains a large amount of polyphenols, and is rich in iodine, vitamins and minerals, and is known to have antioxidant physiological activity.

Crab seaweed and seaweed horsetail are marine flowering plants that contain luteolin, a kind of flavonoid, in their roots and leaf stems, and are known to have antioxidant and anti-inflammatory physiological activities through DPPH radical scavenging activity and SOD-like activity experiments.

Sea urchin is rich in various nutrients and contains large amounts of alpha-tocopherol and carotenoids, which are known to have antioxidant and liver detoxifying physiological activities.

The seaweed extract of the present invention is extracted with any one extraction solvent selected from 70% ethanol or hot water (distilled water). Hereinafter, a method for preparing a seaweed extract having antibacterial activity against Mycobacterium tuberculosis to which the technology of the present invention is applied is described as a solvent It is explained separately for each part and identified based on examples.

[Extraction method using ethanol extraction]

The preferred and verified steps of the method of extracting the seaweed extract having antibacterial activity against Mycobacterium tuberculosis using 70% ethanol extraction solvent are as follows.

(a) In the step of obtaining seaweed powder, seaweed collected from the sea is washed, lyophilized using conventional equipment and methods, and then ground to obtain seaweed powder (S10).

(b) In the step of obtaining the primary seaweed extract, 70% ethanol is added to the seaweed powder, stirred, and then centrifuged to obtain the primary seaweed extract as a supernatant excluding seaweed residues (S20).

(c) In the step of obtaining the secondary seaweed extract, 70% ethanol is added to the seaweed residue, stirred, and then centrifuged to obtain a secondary seaweed extract as a supernatant (S30).

(d) In the step of vacuum concentration, the primary seaweed extract and the secondary seaweed extract are mixed and vacuum concentrated to 70% of the weight of the mixture (S40).

(e) In the step of obtaining the seaweed extract having antibacterial activity against Mycobacterium tuberculosis, the final seaweed extract is obtained by freeze-drying the seaweed extract concentrated above (S50).

[Extraction method using hot water extraction]

Preferred and verified steps of the present invention for a method suitable for extracting a seaweed extract having antibacterial activity against Mycobacterium tuberculosis using a hot water extraction solvent are as follows.

(a) In the step of obtaining seaweed powder, seaweed collected from the sea is washed and lyophilized, and then ground to obtain seaweed powder (S110).

(b) In the step of obtaining the primary seaweed extract, tertiary distilled water is added to the seaweed powder and heated at 90 to 100° C. to obtain the primary seaweed extract (S120).

(c) In the step of obtaining the secondary seaweed extract, after cooling the primary seaweed extract at room temperature, seaweed residues are separated using a sieve, and the secondary seaweed extract is obtained as a supernatant (S130).

(d) In the step of vacuum concentration, the secondary seaweed extract is vacuum-concentrated to 50% by weight (S140).

(e) In the step of obtaining the seaweed extract having antibacterial activity against Mycobacterium tuberculosis, the concentrated seaweed extract above is freeze-dried to obtain the final seaweed extract (S150).

Hereinafter, an embodiment including the present invention will be configured, and the effects thereof will be described in experimental and specific details.

<Example 1 - Seaweed raw material preparation>

36 kinds of seaweeds collected from the East Coast (Pohang, Uljin, Yeongdeok, and Ulleungdo) were thoroughly washed, freeze-dried, and pulverized to obtain powder.

The seaweeds used in the present invention are rhubarb (Ulleungdo), rhubarb (Yeongdeok), seaweed, cap half, streptomycete, buckwheat horse, small spiny cap half, solitary cap half, champagne, agar-agar, dog beet, ringed crab, sticky, Double-headed seosil, Gaeseosil, leaf green, intestine green, crab sea end, leech end, sea urchin, long bulregi end, double-leaved hat half, smooth ring, constricted ring, hoesaeng hat half, mongdang leaf hat half, hibiscus, cow hair red thread It is composed of at least one selected from the group consisting of , reddish black flesh, wrinkled hair, small bead coral end, gaew radish, slippery nuari, holey seaweed, spiny green seaweed, and Sakai stem end.

<Example 2 - Preparation of Seaweed Extract with 70% Ethanol Extraction Solvent>

After adding 250 ml of 70% ethanol as an extraction solvent to 25 g of seaweed raw material powder,

After primary extraction by stirring at 25° C. and 100 to 120 rpm, the supernatant is obtained by centrifugation at 8,000 rpm for 10 minutes.

Secondary extraction was performed under the same conditions by adding 250 ml of 70% ethanol to the residue again.

The primary seaweed extract and the secondary seaweed extract were mixed and concentrated to 70% or more in a vacuum concentrator under reduced pressure, and then lyophilized to recover an ethanol-solvent seaweed extract powder.

<Example 3 - Preparation of Seaweed Extract with Hot Water Extraction Solvent>

2L of tertiary distilled water as an extraction solvent was added to 40g of seaweed raw material powder and heated at about 90 to 100°C for 30 minutes.

After cooling to room temperature, the residue is filtered through a bag-type filter sieve such as a so-called 'Dashi Bag', which is a food filter that is easily available on the market and suitable for application to the present invention, and then the supernatant is obtained. .

After concentrating the supernatant to 50% or more in a vacuum concentrator under reduced pressure, lyophilization was performed to recover the hot-water solvent seaweed extract powder.

<Example 4 - Yield Measurement>

In Example 2 and Example 3, the yields of the seaweed extract powder in ethanol solvent and the seaweed extract powder in hot water solvent obtained relative to the weight of the seaweed raw material powder were measured.

The measurement results are as shown in Table 1 below.

No.

Seaweed raw material name
70% EtOH seaweed extract yield

Hydrothermal Seaweed Extract Yield 20-1 Rhubarb (Ulleungdo) 15.6 32.2 20-2 Rhubarb (Yeongdeok) 5.8 17.5 20-3 Seaweed 438 10.4 20-4 mother and child class 18.6 26.7 20-5 locust 16.0 26.7 20-6 Buchaetmal 17.8 16.7 20-7 half a thorn hat 6.0 12.8 20-8 Lonely Dog Hat Class 6.4 8.3 20-9 true gamble 2.7 7.8 20-10 agar-agar 7.2 8.3 20-11 dog gambling 8.8 12.2 20-12 Ring-jointed crab feet 8.3 16.3 20-13 sticky 8.0 17.3 20-14 double room 12.3 26.4 20-15 writing room 3.5 24.0 20-16 green leaf 8.5 30.4 20-17 blue intestine 4.5 21.5 20-18 Gebada horse 20.8 26.2 20-19 eel horse 18.2 18.7 20-20 sea urchin 19.8 29.7 21-1 end of long trouble 1.3 2.0 21-2 half-leaf hat 5.5 5.8 21-3 smooth hook 3.2 4.0 21-4 narrow hook 2.3 1.0 21-5 hoesaeng hat <0.5 1.1 21-6 Mongdang Leaf Hat 2.3 1.1 21-7 seaweed 6.2 2.6 21-8 cow hair red thread 4.0 12.5 21-9 red black meat <0.5 12.0 21-10 wrinkled hair <0.5 26.8 21-11 small bead coral <0.5 2.5 21-12 Gawoomu <0.5 5.3 21-13 Slippery Nuari 1.3 78.0 21-14 go hole 1.8 28.0 21-15 spiny blue 2.2 8.5 21-16 Sakai big words 3.0 75.0

<Example 5 - Measurement of Mycobacterium tuberculosis activity by MIC50>

The antibacterial activity of Mycobacterium tuberculosis was measured using the ethanol solvent seaweed extract powder and the hot water solvent seaweed extract powder obtained in Examples 2 and 3 above.

Seaweed extract powder was dissolved in DMSO (dimethyl sulfoxide) and used for antibacterial activity.

The antibacterial activity of Mycobacterium tuberculosis was confirmed by measuring the MIC50 (Minimal Inhibitory Concentration, 50% minimum inhibitory concentration for bacterial growth) value, and was measured using Resazurin Microtiter Assay (REMA) for seaweed extract powder.

1) 98 ul of Mueller Hington broth 2 (MH2) medium was added to the first well of a 96-well microtiter plate.

2) 50 ul of medium was added to the remaining wells. About 2 ul of seaweed extract was added to the first well to make the total volume 100 ul. (Initial extract concentration 100 ug/ml)

3) Using the 2-fold serial dilution method, 50 ul of medium from the first well containing the natural product was transferred to the next well using a multi-channel pipette and diluted 2-fold.

4) Mycobacterium tuberculosis mc ² 6230 was prepared to a final concentration of DO ₆₀₀ = 0.005, put into all wells in a volume of 50 ul, covered with a plate, and cultured at 30 ° C for 5 days.

5) Resazurin (0.025% [wt/vol]) was added to each well, and each plate was incubated overnight.

6) The MIC value was calculated using Prism 6 (GraphPad Software, Inc, La Jolla, CA) as, for example, 560/em - 590 nm.

The results of the antibacterial activity of Mycobacterium tuberculosis by MIC50 are shown in Table 2 below.

The control group used bedaquiline (BDQ), a commercially available drug used to treat multidrug-resistant tuberculosis, and the MIC50 (uM) value of betaquiline was 0.29.

No.
MIC50 (μg/ml)
70% EtOH Seaweed Extract Hydrothermal Seaweed Extract 20-1 inactive inactive 20-2 inactive inactive 20-3 inactive inactive 20-4 inactive inactive 20-5 inactive 0.71 20-6 inactive 2.51 20-7 inactive 22.84 20-8 inactive 10.52 20-9 inactive inactive 20-10 inactive inactive 20-11 inactive Not dissolved 20-12 inactive inactive 20-13 inactive Not dissolved 20-14 inactive 22.27 20-15 inactive inactive 20-16 inactive inactive 20-17 27.77 (but inactive) 47.74 20-18 inactive 21.98 20-19 inactive inactive 20-20 inactive inactive 21-1 47.68 inactive 21-2 inactive inactive 21-3 10.27 inactive 21-4 5.09 inactive 21-5 21.28 inactive 21-6 10.43 Not dissolved 21-7 inactive Not dissolved 21-8 inactive inactive 21-9 inactive inactive 21-10 inactive Not dissolved 21-11 Not dissolved Not dissolved 21-12 inactive inactive 21-13 inactive inactive 21-14 inactive inactive 21-15 inactive Not dissolved 21-16 inactive inactive

<Example 6 - Measurement of Mycobacterium tuberculosis activity by RFU>

The antibacterial activity of Mycobacterium tuberculosis was measured using the ethanol solvent seaweed extract powder and hot water solvent seaweed extract powder obtained in Examples 2 and 3 above.

Seaweed extract powder was dissolved in DMSO (dimethyl sulfoxide) and used for antibacterial activity.

The antibacterial activity of Mycobacterium tuberculosis was confirmed by measuring the relative fluorescence unit (RFU) value. The culture method was carried out in the same manner as in Example 5 above.

RFU was measured using a SpectraMax® M3 Multi-Mode Microplate Reader (Molecular Devices, CA, USA) (eg 560/em - 590 nm).

The results of the antibacterial activity of Mycobacterium tuberculosis by RFU are shown in the graph of FIG. 1 below.

As a control group, Bedaquiline (BDQ), a commercially available drug used to treat multidrug-resistant tuberculosis, was used.

As a result of measuring the antibacterial activity of Mycobacterium tuberculosis by MIC50 and RFU using the present invention as described above, hot-water extracts of streptococcus, buchat end, small spiny cap, solitary cap, double-headed seosil, intestine green, crab sea end, and long bullegi end , It was confirmed that the 70% ethanol extract of the smooth-ringed, constricted-ringed, black-headed, and black-headed-leaved caps exhibited higher antibacterial activity against Mycobacterium tuberculosis mc2 6230 than betaquiline.

As described above, the seaweed extract having antibacterial activity against Mycobacterium tuberculosis according to the present invention provides a seaweed extract having an antibacterial activity against Mycobacterium tuberculosis from naturally collected seaweed.

In particular, the present invention is antiviral, antifungal, immunomodulatory, antioxidative, antibacterial, antiviral, antifungal, immunomodulatory, anti-oxidant, anti-tuberculosis, or drug-resistant tuberculosis caused by chemotherapeutic agents such as isoniazid and rifampin, which are conventionally used as anti-tuberculosis agents, or various problems caused by the emergence of drug-resistant strains. It is shown that there is an effect of providing extracts having antibacterial activity against Mycobacterium tuberculosis from 36 kinds of natural seaweeds having various biological activities such as tumor and anti-inflammation.

In addition, the seaweed extract having antibacterial activity against Mycobacterium tuberculosis of the present invention minimizes side effects such as peripheral neuropathy, hepatotoxicity, skin rash, headache, etc. of conventional chemotherapeutic agents, and relieves and improves tuberculosis symptoms without fear of resistance as a natural agent. There is a usefulness that can promote the effect to a considerable extent.

Therefore, it is expected that the seaweed extract having antibacterial activity against Mycobacterium tuberculosis according to the present invention can be widely used in pharmaceuticals, quasi-drugs, and health functional foods for the prevention, improvement, or treatment of tuberculosis.

Not applicable.

Claims (7)

delete delete delete delete Washing seaweed collected from the sea, freeze-drying, and pulverizing to obtain seaweed powder (S10);
Obtaining a primary seaweed extract as a supernatant excluding seaweed residues by adding 70% ethanol to the seaweed powder obtained in the step of obtaining the seaweed powder (S10), stirring, and centrifuging (S20);
Adding 70% ethanol to the seaweed residue from the step of obtaining the first seaweed extract (S20), stirring, and centrifuging to obtain a second seaweed extract as a supernatant (S30);
The first seaweed extract obtained in the step of obtaining the first seaweed extract (S20) and the second seaweed extract obtained in the step of obtaining the second seaweed extract (S30) were mixed and vacuumed to 70% of the weight of the mixture. Concentrating (S40);
Freeze-drying the seaweed extract concentrated in the vacuum concentration step (S40) to obtain a seaweed extract having a growth inhibitory action against Mycobacterium tuberculosis (S50);
In the step of obtaining the seaweed powder (S10), the seaweed is at least one selected from the group consisting of long bule end, smooth round head, constricted end round head, hoesaeng cap, and mongdan leaf cap, Preparation of a seaweed extract having antibacterial activity against Mycobacterium tuberculosis method. Washing seaweed collected from the sea, freeze-drying, and pulverizing to obtain seaweed powder (S110);
obtaining a first seaweed extract by adding tertiary distilled water to the seaweed powder obtained in the step of obtaining the seaweed powder (S110) and heating at 90 to 100° C. (S120);
Cooling the first seaweed extract obtained in the step of obtaining the first seaweed extract (S120) at room temperature, separating the seaweed residues with a sieve, and obtaining a second seaweed extract as a supernatant (S130);
vacuum-concentrating the secondary seaweed extract obtained in the step of obtaining the secondary seaweed extract (S130) to 50% by weight (S140);
Freeze-drying the seaweed extract concentrated in the vacuum concentration step (S140) to obtain a seaweed extract having a growth inhibitory action against Mycobacterium tuberculosis (S150);
In the step of obtaining the seaweed powder (S110), the seaweed has antibacterial activity against Mycobacterium tuberculosis, characterized in that at least one selected from the group consisting of streptococci, spiny worms, small spiny caps, solitary dog caps, double-headed lice, intestinal greens, and crab sea ends. Eggplant seaweed extract manufacturing method. delete

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