KR20170132394A - Pharmaceutical composition containing collinin with antitubercular activity and preparation method for the same - Google Patents

Abstract

The present invention relates to an antituberculous pharmaceutical composition containing a choline chloride isolated from a sanchoan tree or a pharmaceutically acceptable salt thereof, and a method for preparing the antituberculous pharmaceutical composition, wherein the single substance has an antituberculous effect against a Mycobacterium tuberculosis including multidrug resistance and super TB, Or INH, and may be useful as a therapeutic agent for pulmonary tuberculosis.

Description

TECHNICAL FIELD [0001] The present invention relates to an anti-tuberculosis pharmaceutical composition comprising cholinin and a method for producing the same,

The present invention relates to an anti-tuberculosis pharmaceutical composition comprising Collinin and a method for producing the same.

Tuberculosis is a disease that has a history of human history. TB has been confirmed thousands of years ago, and it is one of the oldest diseases that has not been conquered to date.

Although the number of tuberculosis-related deaths has risen to more than 2 million annually worldwide, the development of new therapies has not been developed since the 1960s, and the resistance to drugs due to treatment has become increasingly difficult to treat. Recently, the number of patients with tuberculosis and the number of deaths have declined slightly, but the number of patients with multidrug-resistant and super tuberculosis who are difficult to treat are increasing.

Drug-resistant tuberculosis (MDR-TB) is a type of tuberculosis that is resistant to rifampin and ainas, which are the most basic treatments of tuberculosis drugs. Major causes of tuberculosis are tuberculosis, And the patient is not able to manage the patient sufficiently in many cases.

Super tuberculosis, which is more serious than multidrug-resistant tuberculosis, refers to tuberculosis resistant to both anti-tuberculosis agents rifampin and aiina as well as secondary anti-tuberculosis injections and quinolone antibiotics.

These multidrug-resistant patients and super tuberculosis patients do not have proper therapeutic drugs, and thus serious social problems arise. As a result, patients with tuberculosis may die because of the lack of therapeutic drugs.

It is a global trend to develop drugs using natural products, and it has the advantages of natural pharmaceutical products, which are relatively less toxic than synthetic drugs and shorten development period and costs.

Korean Patent No. 10-1493461, Announcement of Feb. 20, 201, Pharmaceutical Composition for the Treatment of Tuberculosis Using Dioxypilgraminin as an Active Ingredient of White Rice Extract Korean Patent Laid-Open No. 10-2006-0026814, published on Mar. 24, 2006, a pharmaceutical composition for treating tuberculosis and a health functional food

An object of the present invention is to provide an anti-tuberculosis pharmaceutical composition and a method for producing the same using an anti-tuberculosis activity of an effective single component isolated from a natural product.

In order to achieve the above object, a pharmaceutical composition for treating or preventing tuberculosis according to an embodiment of the present invention comprises collinin or a pharmaceutically acceptable salt thereof.

The tuberculosis may include multidrug-resistant tuberculosis and super tuberculosis.

The colinin or a pharmaceutically acceptable salt thereof may have improved anti-TB activity against rifampin (RIF) or isoniazid (INH) against extensively drug-resistant MTB.

The tuberculosis may be pulmonary tuberculosis, tuberculosis tuberculosis, bone tuberculosis, throat tuberculosis, lymph node tuberculosis, breast tuberculosis or spinal tuberculosis.

According to another embodiment of the present invention, there is provided a method for preparing a pharmaceutical composition for treatment or prevention of tuberculosis, comprising the steps of: preparing a crude extract of Panicum japonica extract by immersing the pan plant in an alcohol-containing extraction solvent; A fractionation step of preparing a methylene chloride fraction of an Anguilla japonica extract by applying a fraction solvent containing water and methylene chloride to the Anguilla japonica extract; And an active component separation step of fractionating the methylene chloride fraction by chromatography to obtain a colinin.

Hereinafter, the present invention will be described in more detail.

Throughout the present invention, the term "combination thereof " included in the expression of the machine form means a mixture or combination of one or more elements selected from the group consisting of the constituents described in the expression of the form of a marker, And the like.

In the entirety of the present invention, the description of "A and / or B" means "A, B, or A and B".

Pharmaceutically acceptable salts in the present invention refer to salts which retain the biological effectiveness and properties of the parent compounds and which are not deleterious to the biological or other direction when a single dosage is administered. For example, the above "veterinarily acceptable salt thereof" or "a salt thereof" may be a base addition salt of a specific compound and may be prepared from inorganic and organic bases.

The terms "about "," substantially ", etc. used to the extent that they are used in the present invention are used in their numerical values or in close proximity to their numerical values when the manufacturing and material tolerances inherent in the meanings mentioned are presented, Accurate or absolute numbers are used to help prevent unauthorized exploitation by unauthorized intruders of the referenced disclosure.

The singular < RTI ID = 0.0 > expression < / RTI > in the present invention is interpreted to mean a singular or plural that is to be construed in context unless otherwise stated.

Rice and Zanthoxylumschinifolium Siebold & Zucc.) Is a deciduous shrub that grows about 3m in height. Its fruit is about 5mm in diameter. It grows in October when it gets red. It grows well mainly on the shade of the foot of the mountain, and it is distributed all over the country. Young leaves and fruit are used as a spice for their unique fragrance and sour taste. In Dongbokgop, it warms the efficacy of Sanchodong, makes the taste spicy, makes the hair fall off, keeps the hair from falling, brightens the eyes, It was said to be healing and being a chinchhana. In modern pharmacology, colinin, a quarinine compound isolated from the Sancho wood, has been reported to have some physiological activities such as oral microbial inhibition.

Recently, antituberculous activity against Panax ginseng extract has been reported, but its active ingredient is hydroxy-β-sanshool, which is different from this technology.

The inventors of the present invention, while repeatedly studying a new active substance for anti-tuberculosis derived from a natural product, have succeeded in separating an excellent anti-tuberculosis active substance from an extract of a sancho wood (especially a leaf of a sancho) and found that this substance is resistant to multi- The present inventors completed the present invention by confirming that they exhibit excellent activity in multidrug resistant tuberculosis and super tuberculosis. Specifically, the inventors of the present invention prepared crude extracts using the above-mentioned Sancho tree leaves, and isolated a single compound as an anti-tuberculosis active substance from methylene chloride fractions of fractions thereof. As a result of examining the structure of the active substance, (Collinin). The present invention has been completed based on this finding.

The pharmaceutical composition for treating or preventing tuberculosis of the present invention contains collinin (Formula 1) or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

The above-mentioned pharmaceutically acceptable salt is a salt of the compound represented by the above-mentioned formula (1) which has an effective activity of anti-tuberculosis at a concentration with no or little toxicity. It is an organic or inorganic addition salt of a collinin derivative represented by the formula And includes solvates, hydrates, isomers and the like.

As the inorganic acid, hydrochloric acid, bromic acid, nitric acid, sulfuric acid, perchloric acid, phosphoric acid and the like can be used as the free acid. Examples of the organic acid include citric acid, acetic acid, lactic acid, maleic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, maleic acid, tartaric acid, succinic acid, malonic acid, succinic acid, malonic acid, glutamic acid, aspartic acid, oxalic acid, P-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid or malonic acid. These salts also include alkali metal salts (sodium salts, potassium salts, etc.) and alkaline earth metal salts (calcium salts, magnesium salts, etc.).

For example, the acid addition salt may be selected from the group consisting of acetate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camylate, citrate, eddylate, Hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, malate, glucoside, gluconate, gluconate, glucuronate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / Hydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydroxyacetate, Lactate, stearate, succinate, tartrate, tosylate, trifluoroacetate Diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, zinc salts, and the like. Preferred is hydrochloride or trifluoroacetate.

The acid addition salt according to the present invention can be obtained by a conventional method, for example, by dissolving the collinin represented by Chemical Formula 1 in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile and the like, The resulting precipitate may be filtered and dried. Alternatively, the precipitate may be dried by evaporating the solvent and excess acid under reduced pressure, or may be crystallized in an organic solvent.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or an alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt. The corresponding silver salt can also be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable salt (such as silver nitrate).

The pharmaceutical composition may have anti-tuberculosis activity against various resistant Mycobacterium tuberculosis as well as general tuberculosis. The resistant Mycobacterium tuberculosis may be selected from the group consisting of multidrug-resistant tuberculosis (MDR), extensively drug-resistant tuberculosis (XDR), isoniazid-resistant tuberculosis (INH-r), rifampin-resistant tuberculosis (RIF-r), pyrazinamide- tuberculosis, Strep-r, and combinations thereof.

Particularly, the above-mentioned collinin or a pharmaceutically acceptable salt thereof is effective for treating tuberculosis which is resistant to rifampin (RIF) and isoniazid (INH), which is prescribed as a primary tuberculosis drug, It is highly effective as a novel anti-tuberculosis drug and has advantages over MDR and XDR, which are more effective than existing drugs RIF and INH.

In addition, antitubercular pharmaceutical compositions containing the above-mentioned choline or a pharmaceutically acceptable salt thereof as an active ingredient can be practically used as new medicines and materials according to existing drug resistance.

Furthermore, the above pharmaceutical composition is intended to be used as a therapeutic agent for anti-tuberculosis by isolating an active substance useful for tuberculosis from a natural substance having relatively high stability, and has a merit that it is relatively stable compared to a pre-synthetic drug even though it is a single substance.

The tuberculosis may be pulmonary tuberculosis, tuberculosis, bone tuberculosis, throat tuberculosis, lymphedema tuberculosis, breast tuberculosis or spinal tuberculosis, and may be pulmonary tuberculosis in particular.

The pharmaceutical composition may further comprise a pharmaceutically acceptable carrier or excipient. For example, the pharmaceutical compositions herein may be formulated as tablets, capsules, elixirs, suspensions, syrups, and the like in admixture with conventional pharmaceutical carriers and excipients syrup, and / or wafers, but the present invention is not limited thereto.

The pharmaceutical composition may contain, but is not limited to, about 0.1 to 90% by weight, more typically about 10 to 30%, of active compound.

The pharmaceutical composition may be in the form of corn starch or gelatin, lactose, sucrose, microcrystalline cellulose, kaolin, mannitol, dicalcium phosphate, sodium chloride, and / or alginic acid but are not limited to, common carriers such as alginic acid and excipients.

Croscarmellose, microcrystalline cellulose, corn starch, sodium starch glycolate and / or alginic acid may be used as the disintegrant commonly used in the formulation of the pharmaceutical composition, but are not limited thereto.

In general, the liquid composition may optionally be mixed with a non-aqueous solvent such as, for example, ethanol, glycerin, sorbitol, and polyethylene glycol, together with a suspending agent, a solubilizer (such as cyclodextrin), a preservative, a surfactant, a wetting agent, , A suspension or solution of a compound or a pharmaceutically acceptable salt in a suitable liquid carrier (s) such as an oil or water. On the other hand, liquid formulations may be prepared from, but not limited to, reconstitutable powders. For example, powders containing active compounds, suspensions, sucrose and sweeteners may be reconstituted to form suspensions with water, and syrups may be prepared from powders containing the active ingredient, sucrose and sweetener, .

Compositions in tablet form may be prepared using any suitable pharmaceutical carrier conventionally used in preparing solid compositions. Examples of such carriers may include, but are not limited to, binders such as magnesium stearate, starch, lactose, sucrose, microcrystalline cellulose and polyvinylpyrrolidone. Tablets may also include a color film coating, or may include, but are not limited to, color as a part of the carrier.

Additionally, the active compound may be, but is not limited to, a tablet containing a hydrophilic or hydrophobic matrix and being formulated in a controlled release dosage form.

Compositions in the form of capsules may be prepared, for example, by incorporating the active compounds and excipients into hard gelatine capsules using conventional encapsulation procedures, but are not limited thereto. Alternatively, semi-solid matrices of active compounds and high molecular weight polyethylene glycols can be prepared and filled into hard gelatine capsules, or solutions or edible oils of the active compounds in polyethylene glycol, for example liquids A suspension of paraffin or fractionated coconut oil may be prepared and filled into soft gelatin capsules, but this is not so limited.

Tablet binders include, but are not limited to, acacia, methylcellulose, sodium carboxymethylcellulose, poly-vinylpyrrolidone, hydroxypropylmethylcellulose, sucrose, starch and / or ethylcellulose. Lubricants that may be used include, but are not limited to, magnesium stearate or other metal stearate, stearic acid, silicone fluids, talc, waxes, oils and / or colloidal silicas. have. Fragrances such as peppermint, oil of wintergreen, cherry flavor, and the like may also be used, but may not be limited thereto. In addition, it may be desirable to add a coloring agent to make the dosage form seem more attractive or to help identify the product.

The pharmaceutical composition of the present invention which is activated upon parenteral administration can be formulated for intramuscular, intrathecal, or intravenous administration.

Typical intramuscular or intrathecal dosage compositions may be, but are not limited to, suspensions or solutions of the active ingredient in oils such as arachis oil or sesame oil. Typical intramuscular or submucosal administration compositions may be, for example, but are not limited to, a sterile isotonic aqueous solution containing the active ingredient and dextrose or sodium chloride, or dextrose and sodium chloride. Other examples include, but are not limited to, lactated Ringer's injection, Lactic Ringer & Dextrose Injection, Normosol-M and Dextrose, Isolyte E, . Optionally, a co-solvent such as polyethylene glycol; Chelating agents such as ethylenediaminetetraacetic acid; Antioxidants such as sodium metabisulphite may be included in the formulation, but are not limited thereto. Optionally, the solution may be freeze-dried and reconstituted with a suitable solvent immediately prior to administration, but may not be limited thereto.

The pharmaceutical composition may be in the form of a powder or a solution, but is not limited thereto. The pharmaceutical composition may be orally administered, intraperitoneally injected, intravenously injected, intramuscularly injected, and / or subcutaneously injected, but is not limited thereto.

A method of preparing a pharmaceutical composition for treating or preventing tuberculosis containing collinin according to another embodiment of the present invention comprises the steps of immersing a sancho plant in an extraction solvent containing alcohol, A step of preparing a crude extract of Panax notoginsenoside L.; A fractionation step of preparing a methylene chloride fraction of an Anguilla japonica extract by applying a fraction solvent containing water and methylene chloride to the Anguilla japonica extract; And an active component separation step of fractionating the methylene chloride fraction by chromatography to obtain a colinin.

The step of preparing the crude extract is a step of preparing a crude extract by immersing an anchovy tree in an extraction solvent containing an alcohol. For example, the extract of Zanthoxylumschinifolium Dried pulverized product of leaves, roots, stems, fruits, and combinations thereof can be applied as the sowing wood to be immersed. Specifically, dried and ground pulverized sow leaves can be applied to the sowed leaves of Siebold & Zucc. . The crude extract can be prepared by extracting at room temperature with the extract obtained by immersing it in an extraction solvent. The extraction solvent to be used is an alcohol having 1 to 5 carbon atoms, hexane, chloroform, alcohol, water, May be applied. Specifically, the extraction solvent may be water, methanol, ethanol, alcohol, and mixtures thereof.

The crude extract may be extracted at a extraction temperature of 15 to 100 ° C. for 3 to 5 days using a solvent having a volume of 3 to 20 times the dry weight of the pulverized product. And extracting the liquid phase crude extract in one to five consecutive times.

The sancho wood crude extract may be applied as a crude extract of the liquid phase, as it is, or may be applied as an extract of the sancho wood extract obtained by removing the solid content and concentrating under reduced pressure.

The step of fractionation is a step of recovering the methylene chloride fraction by applying a fraction solvent to the above-mentioned P. acidosis extract. The distilled water and fraction solvent may be applied at a volume ratio of 1: 0.5 to 1.5, and the fractionated solvent may be a mixture of distilled water and distilled water. , N-hexane, methylene chloride, ethyl acetate, n-butanol, and a combination thereof can be applied, and they can be applied sequentially, more specifically, water and methylene chloride can be applied.

The active ingredient separation step is a step of fractionating the methylene chloride fraction by chromatography to obtain a collinin. Specifically, a small fraction was obtained using a mixed solvent concentration gradient of n-hexane-ethyl acetate, and the small fraction was again subjected to reverse phase column chromatography using a mixed solvent of methyl alcohol and water to obtain a single compound, Collinin, .

The pharmaceutical composition for the treatment or prevention of tuberculosis of the present invention has relatively high activity in various tuberculosis strains including tubercle bacilli which are the cause of multi-drug resistant and super tuberculosis, and thus has a high utility as a new anti-tuberculosis drug. Drug-prescribed rifampin and Iina have better antitubercular activity. Furthermore, the above pharmaceutical composition is intended to be used as a therapeutic agent for anti-tuberculosis by isolating an active substance useful for tuberculosis from a natural substance having relatively high stability, and has a merit that it is relatively stable compared to a pre-synthetic drug even though it is a single substance.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily carry out the present invention. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

Example  1: Sancho wood ( Zanthoxylumschinifolium Siebold & Zucc .) Extraction and purification of the colinin from the extract Purified

1-1. Sancho tree leaf extract and Fraction  Produce

4.4 kg of Sancho leaf (Gyeongbuk, Gyeongsan) was extracted with 22 liters of 80% methyl alcohol for 72 hours at room temperature. The extract was filtered using a funnel and filter paper, concentrated under reduced pressure to give 80% methyl alcohol (Hereinafter referred to as " ZS ") dry weight of 1 kg.

1 kg of the crude extract was suspended in 5 L of distilled water and the polar fractions were sequentially sequenced three times in the order of normal hexane, methylene chloride, ethyl acetate and n-butanol. The methylene chloride fraction (hereinafter referred to as "ZSM" ) Was concentrated under reduced pressure to obtain 311.0 g.

1-2. Sancho tree leaves Methylene chloride From the fraction  Separation of compounds

The methylene chloride fractions obtained from 1-1. Of Example 1 were separated into small fractions using open-column chromatography according to the method described below.

Silica gel (230-400 mesh) was suspended and filled in a glass column having a diameter of 9 cm and eluted with normal hexane to obtain a homogeneous state. Then, a sample (311.0 g of methylene chloride fraction of Sancho tree leaf) was adsorbed onto silica gel, Lt; / RTI >

And then subjected to open column chromatography using a mixed solvent concentration gradient of n-hexane-ethyl acetate (n-hexane: ethyl acetate = 100: 1 → 1: 100) to obtain 23 small fractions (hereinafter referred to as "ZSM-1 to ZSM- Name card).

Next, 3.0 g of ZSM-11 fraction was subjected to reverse phase column chromatography (YMC gel ODS-A, 12 nm, S-75 μm) with methyl alcohol / water mixture solvent (4: 6 → 1: And then purified (separation yield of the substance from the herbal medicine: 0.011%).

Example  2: Identification of the structure of isolated compounds

Structural analysis was performed using a Varian Unity INOVA 400 Spectrometer ( ¹ HNMR (400 MHz) and ¹³ C NMR (100 MHz) to determine the structure of the compound isolated and purified in Example 1 above, Results were obtained.

_{^{1 HNMR (CDCl 3, 400MHz)}} ; δ6.24 (1H, d, J = 9.6Hz, H-3), 7.61 (1H, d, J = 9.6 Hz, H-4), 7.13 (1H, d, J = 8.8 Hz, H-5) , 6.86 (1H, d, J = 8.8 Hz, H-6), 4.68 (2H, d, J = 6.4 Hz, H-1 '), 5.48 (1H, t, J = H-2 '), 2.13-2.04 (4H, m, H-4' and H-5 '), 5.07-5.04 8 '), 1.74 (3H, s, H-9'), 1.59 (3H, s, H-10 '), 3.97 (3H, s, -OCH 3); _{^{13 CNMR (CDCl 3, 100MHz)}} ; δ160.9 (C-2), 113.9 (C-3), 143.9 (C-4), 113.6 (C-4a), 122.9 (C-5), 110.4 (C- 6), 155.2 (C-7), 142.0 (C-8), 148.4 (C-9), 66.5 C-4 '), 26.4 (C-5'), 123.9 (C-6 '), 132.1 _{10 '), 61.6 (-OCH 3} )

This compound was identified as a geranyl coumarin-based collinin having the structure of the following formula (1), and it was confirmed that it coincided with data of already known collinin (IS Chen et al., Phytochemistry, 39 (5), 1091-1097, 1995).

[Chemical Formula 1]

Example  3: Identification of anti-tuberculosis activity of compounds

Anti tuberculosis efficacy was measured using REMA (Resazurin microtiter assay) and proceeded as follows.

Resazurin solution was prepared by adding 4 mg of resazurin sodium salt to 40 ml of sterilized water to make 0.01%, then filtered with 0.45 m filter, and stored in the refrigerator. 100 μl of each of the above compounds was added to 96 wells containing 100 μl of 7H9 media (7H9 media), followed by serial dilution (1.5 ㎍ / ml to 50 ㎍ / ml), followed by addition of the tubercle H37Ra (5x10 cells / ml) And H37Rv (5x10 cells / ml), respectively. One week later, 30 res of resazurin solution was added to each well to confirm the anti-tubercular activity. Isoniazid (INH, INA) and rifampin (RIF) were added at 0.5 to 2 / / ml as the positive control, and DMSO was used as the negative control.

MDR (multidrug-resistant MTB, KMRC 00116-00250), extensively drug-resistant MTB, KMRC 00203-00197, rifampin-resistant MTB and KMRC 00121-00341, (Streptomycin-resistant MTB, KMRC 00122-00123), and PZA-r (pyrazinamide-resistant MTB, KMRC 00130-00064) The anti-tuberculosis activity of the compound was confirmed, and the same as above. Isoniazid (INH, INA) and rifampin (RIF) were used as positive control.

The results of the above experiments are summarized in MICs (minimum inhibitory concentrations, μg / ml) and are shown in Table 1 below.

Active compound to be identified MICs (μg / ml) H37Ra H37Rv MDR XDR Examples (Colinin) 6.25-12.5 6.25-12.5 6.25-12.5 6.25-12.5 Positive control (RIF) 0.05-0.1 0.1-0.195 12.5-25 > 100 Positive control (INH) 0.006-0.012 0.006-0.012 25-50 > 100 Active compound to be identified MICs (μg / ml) RIF-r INH-r STR-r PZA-r Examples (Colinin) 12.5-25 6.25-12.5 6.25-12.5 6.25-12.5 Positive control (RIF) 50-100 0.05-0.1 0.05-0.1 0.1-0.195 Positive control (INH) 50-100 > 100 50-100 50-100

As shown in the above Table 1, the compounds of the examples in which anti-tuberculin activity was confirmed in the present invention exhibit relatively uniform anti-tuberculin activity in resistant tuberculosis, multidrug-resistant tuberculosis, and super mycobacteria, , Which is a result of confirming that MDR and XDR are superior to the existing drugs, rifampin and INH, among the anti-tuberculous activities as a single substance of the colinin.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, Of the right.

Claims (5)

A pharmaceutical composition for the treatment or prophylaxis of tuberculosis, which contains Collinin or a pharmaceutically acceptable salt thereof. The method according to claim 1,
Wherein said tuberculosis comprises multidrug-resistant tuberculosis and super tuberculosis. The method according to claim 1,
Wherein said colinin or a pharmaceutically acceptable salt thereof has an anti-tuberculosis activity that is superior to rifampin (RIF) or isoniazid (INH) against extensively drug-resistant MTB. ≪ / RTI > The method according to claim 1,
Wherein the tuberculosis is tuberculosis, tuberculosis, bone tuberculosis, throat tuberculosis, lymphedema tuberculosis, breast tuberculosis or spinal tuberculosis. A step of preparing a crude extract of Panicum japonica extract by immersing the sancho plant in an alcohol-containing extraction solvent;
A fractionation step of preparing a methylene chloride fraction of an Anjunctum saplings extract by applying a fraction solvent containing water and methylene chloride to the Anjungano extract; And
And separating the methylene chloride fraction by chromatography to obtain a colinin,
A method for the preparation of a pharmaceutical composition for the treatment or prophylaxis of tuberculosis, which comprises preparing a pharmaceutical composition for treating or preventing tuberculosis containing Collinin.