KR102552365B1 - Co-amorphous state of curcumin with improved water solubility, and method for manufacturing the same - Google Patents

Abstract

A co-amorphous curcumin complex with poor solubility and a preparation method thereof are disclosed.
The co-amorphous curcumin complex according to the present invention includes a curcumin compound and an amino acid, and has a solubility of 1 μg/ml or more.

Description

Co-amorphous curcumin complex with poor solubility and manufacturing method thereof

The present invention relates to a co-amorphous curcumin complex having poor solubility by combining a curcumin compound with an amino acid, L-lysine, and a preparation method thereof.

Amorphous means a solid state in which molecular interactions exist but no crystalline arrangement is achieved. The amorphous form has a higher energy level than the crystalline form, so it has a higher solubility than the crystalline form. However, due to the high energy level, thermodynamic stability is low, so it has a problem in that the phase transitions to the crystalline form very quickly.

Co-amorphous forms an amorphous solid through an intermolecular bond such as a new hydrogen bond or ionic bond or pi-pi bond between two or more molecules, and can show the effect of suppressing the phenomenon of phase transition to crystalline form. In addition, cavity amorphous is a solid state that can overcome poor solubility through high solubility. The reason why the cavity amorphous form can suppress the phase transition to the crystalline form is that the glass transition temperature of the amorphous form is higher than that of the amorphous form of the drug itself.

The formation of cavity amorphous requires the use of a crystallization method that rapidly reaches a high degree of supersaturation by controlling the crystallization rate very rapidly. Typically, a method of rapidly inducing crystallization rate to a very extreme level, such as vacuum evaporation crystallization, supercritical crystallization, liquid nitrogen crystallization, and freeze-evaporation crystallization, is used.

In this regard, a cocrystal is a form in which two or more different molecules form a crystal structure in a constant stoichiometric ratio.

In the formation of co-crystals, since hydrogen bonds due to interactions between drug molecules and co-molecules are very diverse, it is very difficult to design and control an amorphous form even if this extreme crystallization method is used.

On the other hand, curcumin (Cucumin) is a perennial herb belonging to the ginger family called turmeric (turmeric, turmeric). Curcumin is a yellow-colored spice and is a natural substance contained in 5-10% of the root of a medicinal plant traditionally used to treat inflammation and is cultivated in South and Southeast Asia. Structurally, curcumin is a yellow spice of polyphenols in which two reversible forms of enol-keto interconversion exist.

In addition to curcumin, in turmeric, there are compounds in which the methyl of curcumin's methoxy functional group is missing and two compounds are separated. These curcumin-derived compounds are called curcuminoids. Among them, curcumin shows the best pharmacological activity.

Curcumin is a substance with mainly antioxidant, anti-inflammatory, and action effects, and has been widely used for a long time for medicinal purposes, as a food preservative, and as a natural pigment. And according to current academic reports, curcumin is effective for cardiovascular disease, Alzheimer's disease, metabolic disease, depression, scleroderma, psoriasis, and arthritis. Recently, reports that curcumin is useful for killing various cancer cells are widely known. In addition, curcumin exhibits an effect of promoting blood vessel regeneration, so turmeric extract is sometimes used as a raw material for functional cosmetics.

However, curcumin is classified as a biopharmaceutical classification system (BCS), class IV due to its low water solubility and oral absorption of less than 1%.

Cryst. Growth Des. 2014, 14, 30543061, curcumin has three crystalline forms: form-I, form-II, and form-II. Among them, form-I was reported as the most stable crystalline form. It is then stable in the order of form-II and form-III. Therefore, form-I, the final stable crystalline form, has been marketed and is currently used as a health functional food ingredient.

However, since all three crystalline forms of curcumin are poorly soluble in water, only solubility exists in 40% ethanol aqueous solution. This can be confirmed through Table 1.

[Table 1]

Also, as reported, curcumin form-I, which is currently commercially available, has very low solubility as 0.6 μg/mL.

Therefore, domestic and Japanese hangover cures do not exist in beverages in which curcumin is completely dissolved, and are sold in the form of ingestion/absorption in a suspension state. In addition, since the oral absorption rate of curcumin is less than 1%, it is obvious that the product cannot properly exhibit the effect of relieving a hangover. In addition, there are endless efforts to use curcumin as a pharmaceutical raw material and health functional food all over the world, but it is currently showing limitations in its development due to inability to overcome its poor solubility and oral absorption rate.

Recently, research results on co-crystals of choline alfoscerate and curcumin, and co-crystals of L-carnitine and curcumin have been reported. However, there was no clear improvement in solubility in this study, so no improved results on oral absorption were reported.

In addition, the result of improving the water solubility was insufficient, and only the increase in dissolution through solubility in 40% ethanol-water was reported. In addition, it did not show an increase in oral absorption rate according to an increase in water solubility.

In addition, in an effort to increase the water solubility of curcumin, a study case using clathrate has been reported. However, even with the research results through inclusion, there is a limit to increasing the water solubility of curcumin.

As such, there is currently no case of successfully overcoming the poor solubility of curcumin through a change in the properties of a solid.

An object of the present invention is to provide a co-amorphous curcumin complex having poor solubility.

In addition, an object of the present invention is to use a composition for prevention, a composition for treatment, a food for prevention, a food for treatment, a cosmetic for prevention, or/and a cosmetic for treatment of at least one of antioxidant, anti-inflammatory, arthritis, inflammatory disease, and neoplastic disease. It is to provide a joint amorphous curcumin complex that can be.

Another object of the present invention is to provide a method for preparing a co-amorphous curcumin complex with a simple manufacturing process.

The objects of the present invention are not limited to the above-mentioned objects, and other objects and advantages of the present invention not mentioned above can be understood by the following description and will be more clearly understood by the examples of the present invention. It will also be readily apparent that the objects and advantages of the present invention may be realized by means of the instrumentalities and combinations indicated in the claims.

The co-amorphous curcumin complex according to the present invention includes a curcumin compound and an amino acid, and has a solubility of 1 μg/ml or more.

A method for preparing a co-amorphous curcumin complex according to the present invention includes (a) mixing a curcumin compound, an organic solvent, and an amino acid; (b) heating and stirring the mixed mixture; (c) cooling the stirred mixture; (d) evaporating the organic solvent of the cooled mixture; and (e) vacuum-drying the product of step (d) to obtain a co-amorphous curcumin complex, wherein the co-amorphous curcumin complex has a solubility of 1 μg/ml or more.

The co-amorphous curcumin complex according to the present invention provides an effect with poor solubility.

In particular, the co-amorphous curcumin complex of the present invention shows a value increased by more than 160 times compared to the aqueous solubility of general curcumin.

Accordingly, the co-amorphous curcumin complex according to the present invention is a composition for preventing at least one of antioxidant, anti-inflammatory, arthritis, inflammatory disease, and neoplastic disease, a composition for treatment, a food for prevention, a food for treatment, a cosmetic for prevention, or/ and therapeutic cosmetics are expected to be highly applicable.

In addition to the effects described above, specific effects of the present invention will be described together while explaining specific details for carrying out the present invention.

1 is a photograph of a solubility comparison test to confirm that 30 mg of the Handok ReadyQ content of the co-amorphous curcumin complex and curcumin crystalline form according to the present invention is dissolved in 250 ml of water.
2 and 3 are graphs of the absorption rate in the body of the co-amorphous curcumin complex according to the present invention and general curcumin.
4 is a graph of X-ray diffraction patterns of the co-amorphous curcumin complex powder according to the present invention over time.
5 is an X-ray diffraction pattern graph of the co-amorphous curcumin complex powder according to the present invention.
6 is a differential scanning calorimetry analysis result of the joint amorphous curcumin complex according to the present invention.
FIG. 7 is edisylic acid, FIG. 8 is creatine, FIG. 9 is naphadisylic acid, and FIG. 10 is differential scanning calorimetry analysis results for arginine.

The above objects, features and advantages will be described later in detail with reference to the accompanying drawings, and accordingly, those skilled in the art to which the present invention belongs will be able to easily implement the technical spirit of the present invention. In describing the present invention, if it is determined that the detailed description of the known technology related to the present invention may unnecessarily obscure the subject matter of the present invention, the detailed description will be omitted. Hereinafter, preferred embodiments according to the present invention will be described in detail with reference to the accompanying drawings. In the drawings, the same reference numerals are used to indicate the same or similar components.

In addition, when a component is described as "connected", "coupled" or "connected" to another component, the components may be directly connected or connected to each other, but other components may be "interposed" between each component. ", or each component may be "connected", "coupled" or "connected" through other components. will be.

Hereinafter, a joint amorphous curcumin complex having poor solubility and a preparation method thereof according to some embodiments of the present invention will be described.

In the present invention, a curcumin compound and an amino acid, L-lysine, are combined to provide a co-amorphous curcumin complex with poor solubility. Curcumin's solubility in water is only about 0.6 μg/mL, making it difficult to use it as a raw material for drugs, foods, or health functional foods.

In order to overcome this solubility problem, the co-amorphous curcumin complex of the present invention provides an effect of about 160 times or more increased solubility in water to about 100 μg/mL or more.

The co-amorphous curcumin complex according to the present invention includes a curcumin compound and an amino acid, and has a form in which one molecule of a curcumin compound and one molecule of an amino acid are bonded.

The curcumin compound refers to a compound including curcumin and derivatives thereof. Preferably, the curcumin complex has a form in which curcumin and L-lysine are bound.

In particular, the co-amorphous curcumin complex having such a structure has a solubility of 1 μg/ml or more, preferably 50 μg/ml or more, and more preferably 100 μg/ml or more to 3000 μg/ml or less.

The formation of a cavity amorphous form is not achieved through a very simple and conventional experimental method, but is formed only when the intermolecular interactions between many drug molecules and co-molecules are properly performed. And co-amorphous uses a crystallization method that rapidly controls all crystallization rates to reach supersaturation very quickly or to induce very high supersaturation.

The vacuum evaporation method, which is a crystallization method used in the present invention, is a crystallization method that quickly reaches supersaturation or induces very high supersaturation by rapidly controlling the crystallization rate.

However, even if this crystallization method is used, not all amorphous co-crystals are formed. The reason is that co-crystals are formed through various hydrogen bonds through intermolecular interactions with various conformers, so even if the crystallization rate is rapidly controlled, the change in supersaturation for the formation of an amorphous form is not Each is different.

Therefore, the design and control of cavity amorphous is very complex and requires engineering thoughts and views.

In this regard, in the present invention, a co-crystal with improved water solubility was prepared for the purpose of improving poor solubility. Various conformers were screened to prepare co-crystals. Here, the co-former refers to an inactive molecule among the molecules constituting the crystal of the co-crystal drug.

L-lysine, edisylic acid, napadisilic acid, histidine, aspartic acid, fumaric acid, L-pyroglutamic acid, and citric acid were selected as co-formers and screening was attempted.

As a result, a hollow amorphous curcumin complex was formed only with L-lysine. On the other hand, components such as edisilic acid and naphadisilic acid did not form a hollow amorphous form.

L-lysine has excellent solubility in water and is an amino acid that has both amine (NH ₂ ) and carboxylic acid (COOH) for hydrogen bonding.

In the present invention, a method for reproducibly preparing a co-amorphous curcumin complex using L-lysine was established. The co-amorphous curcumin complex thus prepared showed an effect of increasing water solubility by 166 times or more than that of curcumin.

Since the cavity-amorphous curcumin complex can be sufficiently dissolved in about 250ml of water when ingesting an oral agent on an adult basis, it is possible to predict an increase in absorption of curcumin in the body when the cavity-amorphous curcumin complex is ingested.

That is, the co-amorphous curcumin complex has improved water solubility and can improve absorption in the body, and the co-amorphous curcumin complex has excellent thermodynamic stability because there is no phase transition to a crystalline form.

The co-amorphous curcumin complex of the present invention includes a compound represented by Formula 1 below. And co-amorphous curcumin complex is a novel solid form.

[Formula 1]

The co-amorphous curcumin complex represented by Formula 1 binds 1 to 1.5 equivalents of L-lysine to 1 equivalent of curcumin.

If the amount of L-lysine added exceeds 1.5 equivalents, the amount of solvent used increases 10 to 100 times, and 100% amorphous formation may not be possible.

In the co-amorphous curcumin complex, one molecule of curcumin and one molecule of L-lysine form a co-amorphous form at a ratio of 1:1 by hydrogen bonding.

L-lysine, which has such high water solubility, forms a joint amorphous form due to hydrogen bonding with curcumin. Due to the interaction with L-lysine, when the co-former L-lysine is dissolved, curcumin is also dissolved in water, so the water solubility is increased.

The co-amorphous curcumin complex was dissolved in 250 ml of water with its water solubility increased by about 160 times compared to normal curcumin. This allows the co-amorphous curcumin complex to have an optimized form capable of increasing oral absorption.

The co-amorphous curcumin complex according to the present invention has an amorphous diffraction peak in X-ray diffraction analysis.

In addition, the co-amorphous curcumin complex may be used in at least one preventive composition, therapeutic composition, preventive food, therapeutic food, preventive cosmetic, or/and therapeutic cosmetic of one or more of antioxidant, anti-inflammatory, arthritis, inflammatory disease, and neoplastic disease. It can be included, and its utilization is very high in many fields.

Neoplastic diseases include gastrointestinal stromal tumors (GISTs), small cell lung cancer, non-small cell lung cancer, acute myelocytic leukemia, acute lymphocytic leukemia, and myelodysplastic syndrome ( myelodyplastic syndrome), chronic myelogenous leukemia, colorectal carcinoma, gastric carcinoma, testicular cancer, glioblastoma, astrocytoma or mastocytosis.

The food may be any type of food, such as functional health food, nutritional supplement, nutritional supplement, pharmafood, health food, nutraceutical, designer food, food additive, and the like.

In the present invention, a pharmaceutically useful novel co-amorphous curcumin complex of curcumin is prepared, and a method for preparing a very pure co-amorphous curcumin complex in high yield without performing a separate process of attaching and detaching salts in the manufacturing process. has been established.

In the present invention, the co-amorphous curcumin complex is prepared through the solvent evaporation method, which is a crystallization method. There is a vacuum evaporation method in which vacuum conditions are added to the solvent evaporation method.

The preparation method of the co-amorphous curcumin complex of the present invention is performed through mixing, heating, cooling, evaporation and vacuum drying.

First, a curcumin compound, an organic solvent, and L-lysine as an amino acid are mixed.

After mixing the solid powder curcumin and the organic solvent, L-lysine is added.

Based on the volume of the organic solvent, curcumin may be added at 1 to 100 (w/v)%. Preferably it is 30 to 70 (w / v)%, more preferably it may be added at 40 to 50 (w / v)%.

When preparing the co-amorphous curcumin complex, the co-amorphous curcumin complex was obtained in high yield and an organic solvent effective in removing L-lysine used in excess was used.

The organic solvent includes at least one of methanol, ethanol, isopropyl alcohol, acetone, tetrahydrofuran, dimethyl acetamide, dimethyl sulfoxide, dimethyl formamide, chloroform, methyl ethyl ketone, ethyl acetate, methylene chloride and acetonitrile can do. More preferably, one or more of methanol, ethanol, isopropyl alcohol, and acetone may be included, and most preferably methanol may be included.

1 to 1.5 equivalents of L-lysine as an amino acid are mixed with 1 equivalent of the curcumin compound.

If the equivalent ratio is out of 1 to 1.5 equivalents, it may be difficult to obtain a co-amorphous curcumin complex.

The L-lysine to be added may be added at once, or may be added in separate portions and mixed.

L-lysine may be added to the mixed curcumin compound and the organic solvent in an amount of 1/3 to 2/3. Preferably, it may be divided and added at intervals of 10 to 30 minutes.

The reason why L-lysine is added separately is that when the solid powder curcumin and L-lysine are mixed, the mixing ratio of L-lysine is important in forming an equivalent ratio of 1:1 with curcumin.

Therefore, the co-amorphous curcumin complex can be formed most effectively when 1/3 to 2/3 of L-lysine is divided and mixed rather than adding L-lysine to the solid powder curcumin at once.

Then, the temperature of the mixed mixture was raised to reflux and stirred.

The heating time needs to be adjusted so that it takes more than 1 hour to reach the reflux temperature. The stirring time at reflux temperature should not exceed 3 hours.

Preferably, the temperature is raised for 1 to 3 hours.

Stirring at reflux temperature can be carried out for 1 to 3 hours, preferably for 30 minutes to 2 hours, and more preferably for 30 minutes to 1 hour.

The stirred mixture is then cooled.

The stirred mixture, that is, the refluxed reaction solution is cooled and stirred.

에서 실시하고, 바람직하게는 15~30

에서 실시하며, 보다 바람직하게는 15~20

에서 실시할 수 있다.Cooling is 15~40

Carried out in, preferably 15 to 30

It is carried out in, more preferably 15 to 20

can be carried out in

Stirring is carried out for 1 to 12 hours, preferably for 1 to 8 hours, and more preferably for 3 to 5 hours.

Subsequently, the cooled mixture is stirred while evaporating the organic solvent.

More than 1/2 of the weight of the organic solvent in the cooled mixture may be evaporated.

Evaporation is carried out at 30 to 70 ° C, preferably at 30 to 60 ° C, more preferably at 30 to 50 ° C.

Stirring is carried out for 1 to 12 hours, preferably for 1 to 8 hours, and more preferably for 3 to 5 hours.

In addition, the product obtained by evaporating the organic solvent may be washed with an organic solvent after filtering under reduced pressure.

The organic solvent is at least one of methanol, ethanol, isopropyl alcohol, acetone, tetrahydrofuran, dimethyl acetamide, dimethyl sulfoxide, dimethyl formamide, chloroform, methyl ethyl ketone, ethyl acetate, methylene chloride, and acetonitrile. can include

The organic solvent is added in an amount of 1 to 80 (v/v)%, preferably 30 to 70 (v/v), based on the volume of the organic solvent in the step of mixing the curcumin compound, the organic solvent, and L-lysine as an amino acid. /v)%, more preferably 30 to 50 (v / v)%.

Finally, the product of step (d) is vacuum-dried to obtain a co-amorphous curcumin complex.

The co-amorphous curcumin complex may have a solubility of 1 μg/ml or more.

Vacuum drying can be carried out for 8 to 12 hours at 30 ~ 65 ℃. Preferably vacuum drying is carried out at 40 to 55 ° C, more preferably at 45 to 50 ° C.

The co-amorphous curcumin complex obtained by vacuum drying at the above temperature and time has a water content of 1% or less and a purity of 99.5% or more by HPLC.

As described above, the co-amorphous curcumin complex prepared according to the production method of the present invention is a combination of one molecule of curcumin and one molecule of L-lysine, and is a completely different subject from a simple inclusion of curcumin and L-lysine.

Curcumin incorporation using the currently reported L-lysine polymer has limitations in increasing solubility because it is generated through electrostatic forces between loose molecules.

On the other hand, the co-amorphous curcumin complex of the present invention has more improved water solubility because it is co-amorphous through intermolecular bonds. Accordingly, there is an advantage of improving the absorption of curcumin in the body.

In this regard, the co-amorphous curcumin complex can be used in drugs or health functional foods exhibiting antioxidant and anti-inflammatory effects, and is expected to be very useful.

The specific examples of the co-amorphous curcumin complex with poor solubility and the preparation method thereof are as follows.

[Example 1] Preparation of co-amorphous curcumin complex

After adding 10 g of curcumin and 500 mL of methanol, stir at room temperature for 20 minutes. Thereafter, 1.2 equivalents of L-lysine was added 1/3 each, and the temperature was gradually raised for 1 hour to reach the reflux temperature, followed by further stirring for 30 minutes. After slowly cooling to 20 degrees, the mixture was stirred for 3 hours. Then, while stirring at 50 degrees for 3 hours, 450 mL of methanol was evaporated to precipitate a solid. The precipitated solid was filtered under reduced pressure, washed with 10 mL of isopropyl alcohol, and vacuum-dried at 45 degrees for 16 hours or more to obtain a novel curcumin/L-lysine co-amorphous form (99.2% purity, HPLC) in 90% yield.

[Example 2] Preparation of co-amorphous curcumin complex

After adding 10 g of curcumin and 1,000 mL of ethanol, stir at room temperature for 20 minutes. Thereafter, 1.2 equivalents of L-lysine was added 1/3 each, and the temperature was gradually raised for 1 hour to reach the reflux temperature, followed by further stirring for 30 minutes. After slowly cooling to 20 degrees, the mixture was stirred for 3 hours. Thereafter, 950 mL of ethanol was evaporated while stirring at a temperature of 50 degrees for 3 hours to precipitate a solid. The precipitated solid was filtered under reduced pressure, washed with 10 mL of isopropyl alcohol, and vacuum-dried at 45 degrees for 16 hours or more to obtain a novel curcumin/L-lysine co-amorphous form (99.2% purity, HPLC) in 90% yield.

[Example 3] Preparation of co-amorphous curcumin complex

After adding 10 g of curcumin and 500 ml of acetone, stir at room temperature for 20 minutes. Thereafter, 1.2 equivalents of L-lysine was added 1/3 each, and the temperature was gradually raised for 1 hour to reach the reflux temperature, followed by further stirring for 30 minutes. After slowly cooling to 20 degrees, the mixture was stirred for 3 hours. Thereafter, 450 ml of acetone was evaporated while stirring at 50 degrees for 3 hours to precipitate a solid. The precipitated solid was filtered under reduced pressure, washed with 10 ml of isopropyl alcohol, and vacuum-dried at 45 degrees for 16 hours or more to obtain a new curcumin/L-lysine co-amorphous form (purity: 99.3%, HPLC) with a yield of 88.4%.

[Example 4] Preparation of co-amorphous curcumin complex using evaporation crystallization

After adding 10 g of curcumin and 500 ml of acetone, stir at room temperature for 20 minutes. Thereafter, 1.2 equivalents of L-lysine was added to dissolve all of the mixture, followed by stirring for 2 hours. The stirred solution was completely evaporated using a concentrator (water bath temperature: 50 degrees) to obtain a solid. After adding 30 ml of isopropyl alcohol to the obtained solid, the mixture was stirred for 10 minutes. Thereafter, after filtering under reduced pressure, washing with 10 ml of isopropyl alcohol and vacuum drying at 45 degrees for 16 hours or more to obtain a novel curcumin/L-lysine co-amorphous form (99.15% purity, HPLC) in 94% yield.

[Example 5] Preparation of co-amorphous curcumin complex using evaporation crystallization

After adding 10 g of curcumin and 500 ml of methanol, stir at room temperature for 20 minutes. Thereafter, 1.2 equivalents of L-lysine was added to dissolve all of the mixture, followed by stirring for 2 hours. The stirred solution was completely evaporated using a concentrator (water bath temperature: 50 degrees) to obtain a solid. After adding 30 ml of isopropyl alcohol to the obtained solid, the mixture was stirred for 10 minutes. Thereafter, after filtering under reduced pressure, washing with 10 ml of isopropyl alcohol and vacuum drying at 45 degrees for 16 hours or more to obtain a novel curcumin/L-lysine co-amorphous form (99.15% purity, HPLC) in 94% yield.

[Example 6]

2.0 equivalent of L-lysine was added 1/3 each, and a novel curcumin/L-lysine co-amorphous form (purity 99.2%, HPLC) was obtained in 90% yield under the same conditions as in Example 1.

[Experimental Example 1] Evaluation of solubility of co-amorphous curcumin complex

The water solubility and solubility of the co-amorphous curcumin complex prepared in Examples 1 to 5 and curcumin form-I at pH 6.8 in the intestine were compared and tested.

Analysis conditions are shown in Table 2 below, and the results are summarized in Table 3 below.

Table 2 below is an HPLC test method for solubility analysis of co-amorphous curcumin/L-lysine.

[Table 2]

[Table 3]

As shown in Table 3, it was confirmed that the aqueous solubility of the amorphous curcumin/L-lysine co-crystal and the solubility in the small intestine pH 6.8 were increased by about 160 times higher than that of curcumin.

Curcumin hangover cure product name ‘Ready-Q’ contains 30mg of curcumin.

That is, 30 mg of curcumin was not completely dissolved in 250 ml of water, so the product was in a cloudy state. This is because, as shown in Table 3, only 0.15 mg of curcumin can be dissolved in 250 ml of water.

Additionally, in order to confirm the solubility of the joint amorphous curcumin complex of the present invention, the joint amorphous curcumin complex of the present invention and curcumin were added to a 250 ml Erlenmeyer flask, and the solubility was confirmed, as shown in FIG. 1 .

As a result, it was confirmed that 30 mg of the amorphous curcumin co-crystal of the present invention was all dissolved in 250 ml of water, but curcumin was not.

In addition, with respect to 1 equivalent of the curcumin compound, Example 6 exceeding 1.5 equivalents of amino acids shows a slightly lower water solubility than Examples 1 to 5.

This shows that 1 to 1.5 equivalents of amino acids are advantageous for preparing a curcumin complex with poor solubility.

It has been confirmed that absorption in the body can also be increased by ingesting the co-amorphous curcumin complex with improved solubility of the present invention, and an improvement in absorption rate can be predicted.

[Experimental Example 2] In vivo absorption test of co-amorphous curcumin complex using Caco-2 cell system

The absorption rate test was conducted according to the Caco-2 cell system recommended by the US Food and Drug Administration (FDA).

The Caco-2 permeability assay is a widely adopted and used experimental method that can effectively predict the intestinal absorption rate of many candidate substances (test substances) synthesized in the process of new drug development.

The Caco-2 permeability assay measures the flux rate of a test substance passing through a monolayer of a polarized Caco-2 cell, and uses the generated data to predict drug absorption in vivo.

The Caco-2 cell line is a cell derived from human colon carcinoma and is characterized by the formation of tight junctions between the cells and many microvilli on the apical surface. By evaluating transport in two directions, Apical to Basolateral (A-B) and Basolateral to Apical (B-A) across the cell monomembrane, the efflux ratio, which is an indicator of whether the test substance is active efflux, can be confirmed.

In order to evaluate the intestinal absorption of the co-amorphous curcumin complex, the permeability test of colonocyte Caco-2 monolayer tissue was performed.

Natural curcumin and the co-amorphous curcumin complex prepared in Example 1 were prepared and used at 10 μM and 100 μM, respectively.

The procedure to determine the apparent permeability of curcumin, curcumin complexes across Caco-2 followed the general protocol.

Transport assays were performed in a transwell system at pH 7.4 and 37 degrees.

Before the experiment, the Caco-2 cell monolayer and the top section of the basolateral chamber were washed twice with warmed HBSS solution to remove traces of DMEM and washed. The plate was then incubated with HBSS for 20 minutes at 37 degrees in a CO ₂ incubator, and the TEER value of the monolayer was measured before removing the HBSS. Only cells with an initial TEER value of 300 Ωcm ² or more were used.

In the efflux experiment, a medium containing the prepared sample was placed inside and the monolayer was incubated for 180 minutes. For permeation measurements, aliquots of the culture were released to the apical side of the cells at designated times, and samples were collected again for immediate analysis.

The apparent permeability coefficient (Papp) was calculated by the following formula (dC/dt) × (Vr/AC0). where dc/dt is the flux rate through the monolayer (μg/(mL×s)), Vr is the volume of the receiver chamber (mL), A is the surface area of the cell monolayer and C0 is the initial concentration of the donor fluid (μg/ mL).

[Table 4]

As a result of the absorption rate test according to the test method in Table 4, the co-amorphous curcumin complex provided a curcumin concentration nearly 9.5 times higher than that of normal curcumin in the basolateral aspect (transepithelial transport) as shown in FIG. 2 .

From the results of FIG. 3 , the fact that a larger amount of cavity amorphous curcumin complex was detected in the basolateral region means that it has a very high transport rate compared to normal curcumin.

The absorption concentration was 35.23 μg min ^-1 cm ^-2 for the co-amorphous curcumin complex and 4.98 μg min ^-1 cm ^-2 for normal curcumin, confirming that the co-amorphous curcumin complex was generated due to a higher flux.

Intestinal absorption (Papp) values of the co-amorphous curcumin complex and curcumin were 1.88 × 10 ^-6 cm/s and 0.52 × 10 ^-6 cm/s, respectively.

[Experimental Example 3] Powder X-ray diffraction (PXRD)

PXRD analysis was performed on a (D8 Advance) X-ray powder diffractometer using Cu Kα radiation. The instrument was equipped with tube power, and the amount of current was set to 45 kV and 40 mA. The divergence and scattering slits were set to 1°, and the light-receiving slits were set to 0.2 mm. A θ-2θ continuous scan was used from 5° to 35° 2θ at 3°/min (0.4 sec/0.02° interval).

[Experimental Example 4] Phase transition monitoring through PXRD of co-amorphous curcumin complex powder in water

In order to confirm whether the co-amorphous curcumin complex prepared in Example 1 undergoes a phase transition to a crystalline form, it was sampled every hour while stirring in water for 12 hours, and confirmed through PXRD analysis.

As a result of PXRD analysis and monitoring in FIG. 4 , it was confirmed that the co-amorphous curcumin complex powder did not undergo a phase transition to a crystalline form by 12 hours and was thermodynamically stable.

Therefore, it was confirmed that the co-amorphous curcumin complex is a thermodynamically stable amorphous form.

Therefore, the joint amorphous curcumin complex of the present invention is a new composition that overcomes the poor solubility, which is a problem of curcumin, and is expected to be a new improved drug, food, and health functional food raw material.

[Comparative Example 1] Screening of co-amorphous curcumin complex

L-lysine, edisylic acid, napadisilic acid, histidine, aspartic acid, fumaric acid, L-pyroglutamic acid, and citric acid, which are conformers, were selected and screened.

As a result, it was confirmed through the PXRD pattern of FIG. 5 and the calorimetry curve of differential scanning calorimetry in FIG. 6 that curcumin co-amorphous was formed only in the L-lysine of the present invention.

FIG. 7 is edisylic acid, FIG. 8 is creatine, FIG. 9 is naphadisylic acid, and FIG. 10 is differential scanning calorimetry analysis results for arginine.

Referring to FIGS. 7 to 10 , all but L-lysine could not form a joint amorphous form.

As described above, the present invention has been described with reference to the drawings illustrated, but the present invention is not limited by the embodiments and drawings disclosed in this specification, and various modifications are made by those skilled in the art within the scope of the technical idea of the present invention. It is obvious that variations can be made. In addition, although the operational effects according to the configuration of the present invention have not been explicitly described and described while describing the embodiments of the present invention, it is natural that the effects predictable by the corresponding configuration should also be recognized.

Claims (11)

It is a combined form of a curcumin compound and an amino acid,
The amino acid includes L-lysine,
Co-amorphous curcumin complex having a solubility of 1 μg/ml or more.
According to claim 1,
A joint amorphous curcumin complex in which 1 to 1.5 equivalents of amino acids are bound to 1 equivalent of the curcumin compound.
According to claim 1,
Co-amorphous curcumin complex in which the diffraction peak has an amorphous form in X-ray diffraction analysis.
According to claim 1,
Antioxidant, anti-inflammatory, arthritis, inflammatory diseases, co-amorphous curcumin complex contained in at least one preventive composition, therapeutic composition, preventive food, therapeutic food, preventive cosmetic or / and therapeutic cosmetic of one or more of neoplastic diseases.
(a) mixing a curcumin compound, an organic solvent, and an amino acid;
(b) heating and stirring the mixed mixture;
(c) cooling the stirred mixture;
(d) evaporating the organic solvent of the cooled mixture; and
(e) vacuum-drying the product of step (d) to obtain a co-amorphous curcumin complex in which a curcumin compound and an amino acid are bonded;
The amino acid includes L-lysine,
The method of preparing a joint amorphous curcumin complex, wherein the joint amorphous curcumin complex has a solubility of 1 μg / ml or more.
According to claim 5,
In step (a), 1 to 1.5 equivalents of amino acids are mixed with respect to 1 equivalent of the curcumin compound.
According to claim 5,
In the step (c), a method for preparing a joint amorphous curcumin complex, which is cooled at 15 to 40 ° C. for 1 to 8 hours.
According to claim 5,
In step (d), at least 1/2 of the weight of the organic solvent in the cooled mixture is evaporated.
According to claim 5,
The organic solvent is at least one of methanol, ethanol, isopropyl alcohol, acetone, tetrahydrofuran, dimethyl acetamide, dimethyl sulfoxide, dimethyl formamide, chloroform, methyl ethyl ketone, ethyl acetate, methylene chloride, and acetonitrile. A method for producing a co-amorphous curcumin complex comprising:
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