Patent Application
20200009211
The present invention relates to a curcumin-containing preparation and the like.
Various studies have confirmed the safety and efficacy of curcumin. It has also been reported that curcumins are considered to have various physiological effects, such as suppression of cholesterol elevation, suppression of blood pressure elevation, suppression of blood glucose elevation, anti-allergy, suppression of body fat, and the like.
To expect such physiological effects, the ingestion of large amounts of curcumin is necessary.
Curcumins are components contained in, for example, edible plants. Although curcumins can be ingested, for example, in usual meals, ingesting curcumins in the form of curcumin-containing tablets or like solid preparations is convenient and efficient.
However, most curcumins are poorly soluble in water. Therefore, even if a curcumin-containing solid preparation is ingested, curcumins are dissolved and absorbed into the body fluid at slow rates.
To solve this problem, for example, Patent Document 1 suggests a preparation for oral administration comprising a curcuminoid and an essential oil of turmeric.
Further, several strategies have been attempted to improve the bioavailability of curcumin, such as regulation of curcumin administration route and media, blockade of metabolic pathways by co-administration with other drugs, modification of the bond and structure of curcumin, and the like.
Despite these attempts, the use of curcumin for human health is actually limited. This strongly suggests the need for further improvements in the bioavailability of curcumin.
Therefore, the development of a new technique has been desired, in view of the efficient ingestion of curcumins.
Patent Document 1: JP2012-188450A
An object of the present invention is to provide a preparation for treating or preventing diseases or symptoms that benefit from curcumin absorption into cells.
The inventors of the present invention carried out extensive research to solve the problems, and found that efficient curcumin ingestion (incorporation into cells etc.) becomes possible by a preparation comprising a solid composition comprising:
Based on this finding, the inventors also found that the problems can be solved by this preparation, and completed the present invention.
The present invention encompasses the following aspects.
A preparation for treating or preventing a disease or symptom that benefits from curcumin absorption into cells, the preparation comprising a solid composition comprising:
The preparation according to item 1, wherein the disease or symptom is at least one member selected from the group consisting of:
The preparation according to item 1, wherein the treatment or prevention of a disease or symptom is at least one member selected from the group consisting of:
The preparation according to item 1, wherein the treatment or prevention of a disease or symptom is at least one member selected from the group consisting of suppression of cholesterol elevation, suppression of triglyceride elevation, suppression of chylomicron elevation, suppression of blood pressure elevation, suppression of blood glucose elevation, anti-allergy, and suppression of body fat.
The preparation according to any one of items 1 to 4, wherein the hydrophilic polymer is at least one member selected from the group consisting of polyvinylpyrrolidone, hydroxypropyl cellulose, and hydroxypropyl methylcellulose.
The preparation according to any one of items 1 to 5, wherein the nonionic surfactant is a polyglycerol fatty acid ester.
The preparation according to any one of items 1 to 6, wherein the preparation is a preparation for oral administration, a preparation for gastrointestinal administration, a preparation for transdermal administration, or a preparation for pulmonary administration.
The preparation according to any one of items 1 to 7, wherein the preparation is a pharmaceutical product, quasi-drug, health food, food with function claims, dietary supplement, food with nutrient function claims, nutritional supplement, food for special dietary use, or a food for specified health use.
A method for treating or preventing a disease or symptom that benefits from curcumin absorption into cells, the method comprising administering a solid composition comprising:
to a subject in need thereof.
The method according to item Al, wherein the disease or symptom is at least one member selected from the group consisting of:
The method according to item A1, wherein the treatment or prevention of a disease or symptom is at least one member selected from the group consisting of:
The method according to item A1, wherein the treatment or prevention of a disease or symptom is at least one member selected from the group consisting of suppression of cholesterol elevation, suppression of triglyceride elevation, suppression of chylomicron elevation, suppression of blood pressure elevation, suppression of blood glucose elevation, anti-allergy, and suppression of body fat.
The method according to any one of items A1 to A4, wherein the hydrophilic polymer is at least one member selected from the group consisting of polyvinylpyrrolidone, hydroxypropyl cellulose, and hydroxypropyl methylcellulose.
The method according to any one of items A1 to A5, wherein the nonionic surfactant is a polyglycerol fatty acid ester.
The method according to any one of items A1 to A6, wherein the administration is oral administration, administration through gastrointestinal tract, transdermal administration, or pulmonary administration.
The method according to any one of Items A1 to A7, wherein a pharmaceutical product, quasi-drug, health food, food with function claims, dietary supplement, food with nutrient function claims, nutritional supplement, food for special dietary use, or a food for specified health use is administered in the method.
A solid composition for treating or preventing a disease or symptom that benefits from curcumin absorption into cells, the solid composition comprising:
The solid composition according to item B1, wherein the disease or symptom is at least one member selected from the group consisting of:
The solid composition according to item B1, wherein the treatment or prevention of a disease or symptom is at least one member selected from the group consisting of:
The solid composition according to item B1, wherein the treatment or prevention of a disease or symptom is at least one member selected from the group consisting of suppression of cholesterol elevation, suppression of triglyceride elevation, suppression of chylomicron elevation, suppression of blood pressure elevation, suppression of blood glucose elevation, anti-allergy, and suppression of body fat.
The solid composition according to any one of items B1 to B4, wherein the hydrophilic polymer is at least one member selected from the group consisting of polyvinylpyrrolidone, hydroxypropyl cellulose, and hydroxypropyl methylcellulose.
The solid composition according to any one of items B1 to B5, wherein the nonionic surfactant is a polyglycerol fatty acid ester.
The solid composition according to any one of items B1 to B6, wherein the solid composition is a solid composition for oral administration, a solid composition for gastrointestinal administration, a solid composition for transdermal administration, or a solid composition for pulmonary administration.
The solid composition according to any one of items B1 to B7, wherein the solid composition is a pharmaceutical product, quasi-drug, health food, food with function claims, dietary supplement, food with nutrient function claims, nutritional supplement, food for special dietary use, or a food for specified health use.
A composition for the manufacture of a preparation for treating or preventing a disease or symptom that benefits from curcumin absorption into cells, the composition comprising a solid composition comprising:
The composition according to item Cl, wherein the disease or symptom is at least one member selected from the group consisting of:
The composition according to item C1, wherein the treatment or prevention of a disease or symptom is at least one member selected from the group consisting of:
The composition according to item C1, wherein the treatment or prevention of a disease or symptom is at least one member selected from the group consisting of suppression of cholesterol elevation, suppression of triglyceride elevation, suppression of chylomicron elevation, suppression of blood pressure elevation, suppression of blood glucose elevation, anti-allergy, and suppression of body fat.
The composition according to any one of items C1 to C4, wherein the hydrophilic polymer is at least one member selected from the group consisting of polyvinylpyrrolidone, hydroxypropyl cellulose, and hydroxypropyl methylcellulose.
The composition according to any one of items C1 to C5, wherein the nonionic surfactant is a polyglycerol fatty acid ester.
The composition according to any one of items C1 to C6, wherein the preparation is a preparation for oral administration, a preparation for gastrointestinal administration, a preparation for transdermal administration, or a preparation for pulmonary administration.
The composition according to any one of items C1 to C7, wherein the preparation is a pharmaceutical product, quasi-drug, health food, food with function claims, dietary supplement, food with nutrient function claims, nutritional supplement, food for special dietary use, or a food for specified health use.
When orally administered or ingested, the preparation of the present invention allows curcumin to dissolve at a high rate into the body fluid (preferably the intestinal fluid) for a prolonged period of time, to thereby enable efficient ingestion of the curcumin.
The preparation of the present invention more easily enables curcumin contained therein to be incorporated into cells.
More specifically, the present invention makes it possible to provide a curcumin-containing preparation that enables efficient ingestion of curcumin.
Further, such a curcumin-containing preparation also enables provision of an excellent composition for treating or preventing diseases or symptoms that benefit from curcumin absorption into cells.
The symbols and abbreviations used in this specification can be assumed to have their ordinary meanings used in the technical field to which the present invention pertains, as understood from the context of the specification, unless otherwise specified.
In the specification, the terms “containing” and “comprising” are used to include meanings of the phrase “consisting essentially of” and the phrase “consisting of.”
The step, treatment, or operation disclosed in the specification can be performed at room temperature, unless otherwise specified. In this specification, room temperature refers to a temperature in the range of 10 to 40° C.
The preparation of the present invention comprises a solid composition containing:
The preparation of the present invention encompasses a preparation essentially consisting of the solid composition, and a preparation consisting of the solid composition.
In general, curcumins are crystalline, and are thus poorly soluble in water or insoluble in water.
“Poorly water-soluble” as used herein specifically means that the solubility in pure water is 0.1 mass % or less at 25° C., or that the octanol/water partition coefficient (logP) falls within the range of −1.0 to 4.0. The logP value can be determined by high-performance liquid chromatography according to JIS Z 7260-117 (2006).
The logP value is defined by the following formula:
logP=log(Coc/Cwa)
Coc: concentration of the test substance in the 1-octanol layer
Cwa: concentration of the test substance in the aqueous layer
The “curcumin” used in the present invention may have a solubility of 0.2 mg/100 mL or less with respect to the second dissolution medium of the Japanese Pharmacopoeia, 16th edition, which is determined in accordance with the method prescribed in the Japanese Pharmacopoeia dissolution test.
The curcumin contained in the solid composition may be, for example, an extract derived from a natural product (e.g., turmeric extract), or a synthetic product.
The curcumin contained in the solid composition may have a keto form, an enol form, or a mixture thereof.
The curcumin content in the solid composition is preferably within the range of 1 to 60 mass %, more preferably within the range of 5 to 50 mass %, further preferably 7 to 40 mass %, and even more preferably within the range of 10 to 35 mass %.
Although the solid composition may contain crystalline curcumin, the amount or the proportion of the crystalline curcumin relative to the entire solid composition or total curcumins is preferably small.
The amorphous state of the curcumin contained in the solid composition can be confirmed by powder X-ray diffraction, differential scanning calorimetry, or like methods. Further, the amount of the amorphous curcumin can be calculated from the peak areas of differential scanning calorimetry.
It is particularly preferable that the solid composition is substantially or entirely free of crystalline curcumin. In other words, the curcumin contained in the solid composition of the present invention is preferably substantially amorphous.
The amount of total curcumins in the solid composition (the “total curcumins” include curcumins and crystalline curcumins) is preferably within the range of 1 to 60 mass %, more preferably within the range of 5 to 50 mass %, further preferably 7 to 40 mass %, and even more preferably within the range of 10 to 35 mass %.
The hydrophilic polymer used in the present invention is not necessarily hydrophilic or water-soluble under every condition. The hydrophilic polymer is preferably hydrophilic or water-soluble at least at the pH in the intestinal tract.
The hydrophilic polymer used in the present invention is preferably a solid at room temperature.
The hydrophilic polymer used in the present invention preferably has a glass transition temperature (Tg) of preferably about 50° C. or more, more preferably about 80° C. to about 180° C. The determination of the glass transition temperature (Tg) can be performed according to JIS K 7121: 2012.
The solid composition may contain one hydrophilic polymer, or two or more hydrophilic polymers.
Examples of the hydrophilic polymer used in the present specification include the followings.
In one preferable embodiment of the present invention, the solid composition may contain, as the hydrophilic polymer, at least one member selected from the group consisting of polyvinylpyrrolidone, hydroxypropyl cellulose, and hydroxypropyl methylcellulose; and may further contain other hydrophilic polymers.
In one particularly preferable embodiment of the present invention, the solid composition may contain at least a polyvinylpyrrolidone as the hydrophilic polymer, and may further contain other hydrophilic polymers.
In another preferable embodiment of the present invention, the hydrophilic polymer is at least one member selected from the group consisting of polyvinylpyrrolidone, hydroxypropyl cellulose, and hydroxypropyl methylcellulose.
In another particularly preferable embodiment, the hydrophilic polymer is-polyvinylpyrrolidone.
The hydrophilic polymer content of the solid composition is preferably within the range of 5 to 90 mass %, more preferably within the range of 20 to 90 mass %, and even more preferably within the range of 40 to 90 mass %.
The nonionic surfactant contained in the solid composition is a nonionic surfactant that is at least one member selected from the group consisting of polyglycerol fatty acid esters, sucrose fatty acid esters, and lecithins.
Examples of polyglycerol fatty acid esters used in the present invention include esters of (a) polyglycerols having an average degree of polymerization of 2 or more (preferably 3 to 15, more preferably 3 to 10), and (b) fatty acids having 8 to 18 carbon atoms (e.g., caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, and linoleic acid).
Specific examples of polyglycerol fatty acid esters used in the present invention include diglycerol monolaurate, diglycerol monostearate, diglycerol monooleate, decaglycerol monolaurate, decaglycerol monostearate, and decaglycerol monooleate.
In the present invention, the polyglycerol fatty acid esters can be used singly, or in a combination of two or more.
The HLB value of sucrose fatty acid esters used in the present invention is preferably 5 or more, more preferably 7 or more, further preferably 10 or more, and further more preferably 12 or more.
The fatty acid of the sucrose fatty acid ester used in the present invention preferably has at least 12 carbon atoms, and more preferably 12 to 20 carbon atoms.
Specific examples of sucrose fatty acid esters preferably used in the present invention include sucrose laurate, sucrose myristate, sucrose palmitate, sucrose stearate, sucrose oleate, sucrose behenate, and sucrose erucate.
In the present invention, the sucrose fatty acid esters can be used singly, or in a combination of two or more.
The lecithin used in the present invention is an adduct of a phosphoric acid derivative of di-fatty acid ester of glycerol (diglyceride). Lecithin is widely distributed in plant and animal bodies.
Examples of the lecithin used in the present invention include egg yolk lecithin contained in egg yolk, soybean lecithin contained in soybeans, and sunflower lecithin contained in sunflowers.
Examples of the lecithin used in the present invention include fractionated lecithin obtained by extracting an active ingredient from a lecithin described above, enzymatically modified lecithin obtained by treating lecithin with an enzyme, and enzymatically decomposed lecithin.
Specific examples of the lecithin used in the present invention include lecithin, enzymatically decomposed lecithin (phosphatidic acid), lysolecithin, soybean lecithin (soybean phospholipid), and egg yolk lecithin.
Lecithins that can be used in the present invention are commercially available. For example, SLP-White (trade name, produced by Tsuji Oil Mill Co., Ltd.) can be used.
In the present invention, lecithins can be used singly, or in a combination of two or more.
Particularly suitable examples of the nonionic surfactant contained in the solid composition include polyglycerol fatty acid esters.
The solid composition may contain one or more nonionic surfactants.
In a preferred embodiment of the present invention, the nonionic surfactant is a polyglycerol fatty acid ester.
The nonionic surfactant content in the solid composition is preferably within the range of 5 to 90 mass %, more preferably within the range of 5 to 60 mass %, and further preferably within the range of 10 to 40 mass %.
If necessary, the solid composition may contain components other than those mentioned above, as long as the effects of the present invention are not significantly impaired.
Examples of such components include excipients, fillers, extenders, binders, disintegrators, surfactants, seasonings, flavoring agents, and lubricants.
As long as the effect of the present invention is not significantly impaired, the types and amounts of such components may be suitably selected and designed based on common general technical knowledge.
The preparation of the present invention may be used as a pharmaceutical product, quasi-drug, health food, food with function claims, dietary supplement (supplement), food with nutrient function claims, nutritional supplement, food for special dietary use, a food for specified health use, or the like.
The preparation of the present invention may be a preparation for oral administration, a preparation to be applied to the oral cavity, a preparation to be applied to bronchus and lung, a preparation to be applied to eyes, a preparation to be applied to ears, a preparation to be applied to nose, a preparation to be applied to rectum, a preparation to be applied to vagina, or a preparation to be applied to skin.
Further, the preparation of the present invention may preferably be a preparation for oral administration, a preparation for gastrointestinal administration, a preparation for transdermal administration, or a preparation for pulmonary administration; and more preferably a preparation for oral administration.
Examples of suitable forms of the preparation include tablets (e.g., orally disintegrating tablets, chewable tablets, effervescent tablets, dispersible tablets, soluble tablets), capsules, granules (e.g., effervescent granules), powdered drug, liquids and solutions for oral administration (e.g., elixirs, suspensions, emulsions, limonades), syrups (e.g., preparations for syrups), jellies for oral administration, tablets for oro-mucosal application (e.g., troches, sublingual tablets, buccal tablets, mucoadhesive tablets, medicated chewing gum), sprays for oro-mucosal application, semi-solid preparations for oro-mucosal application, preparations for gargle, dialysis agents (e.g., peritoneal dialysis agents, hemodialysis agents), inhalant solutions (e.g., dry powder inhalers, inhalation solutions, inhalation aerosols), suppositories, semi-solid preparations for rectal application, enemas for rectal application, ophthalmic ointments, ear preparations, nasal preparations (e.g., nasal dry powder inhalers, nasal liquids and solutions), vaginal tablets, vaginal suppositories, solid preparations for cutaneous application (e.g., powders for cutaneous application), liquids and solutions for cutaneous application (e.g., liniments, lotions), and sprays (e.g., aerosols for cutaneous application, pump sprays).
When the preparation is a cosmetic or a similar product, examples of suitable forms thereof include aqueous lotions (e.g., lotions), emulsions, and creams.
Further, examples of the preparations of the present invention include dental care products (e.g., toothpaste) and oral care products (e.g., mouthwash).
These preparations may be produced based on common technical knowledge related to the manufacture of preparations containing a solid composition or the manufacture of preparations in the form of a solid composition, according to the dosage form of the preparation.
The solid composition can be used as a material for producing such a preparation.
The content of the solid composition in the preparation of the present invention may vary depending on the dosage form of the preparation.
For example, the lower limit thereof may be 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 mass %.
For example, the upper limit thereof may be 20, 30, 40, 50, 60, 70, 80, 90, or 100 mass %.
The content may be, for example, 10 to 90 mass %, 20 to 80 mass %, 30 to 70 mass %, or 40 to 60 mass %.
The amount of the preparation (e.g., oral curcumin preparation) of the present invention to be administered or ingested may vary according to the age, body weight, and condition of the user; form of administration; treatment period; and the like. For example, the WHO Technical Report series shows that the ADI of curcumin is 0 to 3 mg/body weight (kg)/day, and that the NOAEL of curcumin is 250 to 320 mg/body weight (kg)/day (WHO Technical Report Series: 1237259778265_0.pdf, page 33). The preparation in an amount within this range can be preferably administered or ingested once a day, or in divided doses several times a day (e.g., twice, three times, four times, or five times).
The solid composition obtained by the present invention can be used not only for pharmaceutical products, foods, or the like, but can also be used by incorporation into, for example, cosmetics. Examples of the form of such cosmetics include skin care cosmetics such as lotions, creams, skin lotions, emulsions, and serums; hair care products, such as shampoos; mouthwashes; and the like. Further, any components that are commonly used in the field of cosmetics can be used in combination.
Examples of surfactants include anionic surfactants such as glycerol fatty acid esters, propylene glycol fatty acid esters, sorbitan fatty acid esters, polyoxyethylene alkyl ethers, polyoxyethylene alkyl phenyl ethers, polyoxyethylene fatty acid esters, carboxylic acid salts, and sulfonic acid salts; and cationic surfactants such as amine salts and ammonium salts. For example, at least one of such surfactants can be used in combination with the solid composition of the present invention.
When administered or ingested orally or via the gastrointestinal tract, the solid composition of the present invention exhibits a high ability to dissolve curcumin into the aqueous medium (e.g., body fluid such as gastric fluid, intestinal fluid, or saliva) in the body for a prolonged period of time, thus enabling efficient ingestion of the curcumin.
In other words, the composition of the present invention is exposed to the aqueous medium (e.g., body fluid such as gastric fluid, intestinal fluid, or saliva) in the body, and the curcumin contained in the composition is highly absorbed to the cells of the body via the medium.
The solid composition of the present invention may preferably be used to treat or prevent diseases or symptoms that benefit from curcumin absorption into cells.
In this specification, the term “treatment” may also mean alleviation of the symptoms that have already occurred.
In this specification, the term “prevention” may also mean alleviation of the symptoms that may occur.
The diseases or symptoms of the present invention may be at least one member selected from the group consisting of:
The diseases or the symptoms of the present invention may be at least one member selected from the group consisting of:
Examples of the hangover symptoms after alcohol ingestion include nausea, headache, and stomach discomfort, as generally understood.
The treatment or prevention of the diseases or the symptoms of the present invention may be at least one member selected from the group consisting of: suppression of cholesterol elevation, suppression of triglyceride elevation, suppression of chylomicron elevation, suppression of blood pressure elevation, suppression of blood glucose elevation, anti-allergy, and suppression of body fat.
The solid composition can be produced, for example, through a method comprising mixing:
In the mixing step, the above components can be mixed simultaneously or successively.
The mixing step can be preferably performed without using a solvent such as an organic solvent.
Even when the solvent is used, the above components, such as curcumin, do not have to be completely dissolved in the solvent.
This allows the composition of the present invention to be produced at low cost, without using a large container or the like.
The step of mixing the components and the step of converting the crystalline curcumin to amorphous curcumin can be separate steps, or they can be partially or completely in common.
A higher conversion of crystalline curcumin to amorphous curcumin is preferable. Converting all or substantially all of the crystalline curcumin to amorphous curcumin is particularly preferable.
The solid composition can be produced, for example, by solvent precipitation methods, spray-drying methods, freeze-drying methods, drying under reduced pressure, or kneading methods; or a combination of these methods.
The solid composition is preferably produced by a production method comprising the step of kneading:
In the kneading step, the crystalline curcumin, the hydrophilic polymer, and the nonionic surfactant are preferably kneaded simultaneously.
The kneading converts a part of the crystalline curcumin to amorphous curcumin, or preferably converts all or substantially all of the crystalline curcumin to amorphous curcumin.
The kneading can be preferably performed, for example, by using a single-screw extruder, an intermeshing screw extruder, or a multi-screw extruder (e.g., a twin-screw extruder). The kneading can also be preferably performed by kneading with a relatively weak force, such as kneading by hand using a spatula or the like on a hot plate.
In this kneading, for example, the mixture is kneaded while heated to the temperature at which the components are dissolved; then, after the components are dissolved, the mixture is cooled to room temperature. The resulting solid composition is pulverized into a powder using a pulverizer to obtain the composition of the present invention.
The primary particle diameter of the solid composition may be appropriately selected according to the form of the preparation of the present invention.
For example, the lower limit of the primary particle diameter of the solid composition may be 0.1 μm, 0.5 μm, 1 μm, 5 μm, 10 μm, 50 μm, or 100 μm.
For example, the upper limit of the primary particle diameter of the solid composition may be 0.5 μm, 1 μm, 5 μm, 10 μm, 50 μm, 100 μm, or 200 μm.
For example, the primary particle diameter may be in the range of 0.1 to 500 μm, 0.5 to 500 μm, 0.5 to 200 μm, 1 to 100 μm, or 10 to 100 μm.
In the preparation of the present invention, the primary particles of the solid composition may constitute secondary particles or the preparation itself according to the dosage form of the preparation, as can be generally understood by a person skilled in the art.
Further, in the preparation of the present invention, the solid composition may not be in the form of particles; and may be in the form of, for example, an uniform tablet, as can be generally understood by a person skilled in the art.
The solid composition is preferably produced by, for example, a method comprising the steps of:
Examples of drying methods include spray-drying methods, freeze-drying methods, vacuum-drying methods, drum-drying methods, far-infrared drying methods, and the like. Spray-drying methods are particularly preferable.
The present invention is described in more detail below with reference to Examples. However, the scope of the present invention is not limited to these Examples.
The symbols and abbreviations in the Examples are defined as follows.
In the Examples, fine granules refer to fine-grained agents, as described in the Japanese Pharmacopoeia, 17th edition; i.e., a preparation of which the total amount passes through a No. 18 (850 μm) sieve, and of which 10% or less of the total amount remains on a No. 30 (500 μm) sieve.
In the Examples, “%” can be understood to mean mass %, based on common technical knowledge and the context, unless otherwise specified.
Compositions having the formulations shown below in Table 1 were individually kneaded with heating to the melting temperature. After melting, each composition was cooled to room temperature, and formed into a powder using a pulverizer. The powder thus obtained was used. However, in the preparation of the composition of Comparative Example 3, the components were merely mixed without heating, and the resulting mixture was used as a test sample.
The kneading with heating was performed by setting a hot plate at 240° C., and kneading each composition by hand using a spatula or the like until the composition was melted.
The components used in the Examples or Comparative Examples are described below. Except for PGFE(A), commercially available products were purchased and used. PGFE(A) is a polyglycerol myristic acid ester of HLB12.
Curcumin material (purity: containing 90% or more curcumin, 4% or more bisdemethoxycurcumin, and 0.1% or more demethoxycurcumin) (bulk powder)
Kollidon K30 (trade name, BASF A.G.): PVP (polyvinylpyrrolidone)
PGFE(A): PGFE (polyglycerol fatty acid ester)
Ryoto Polyglyester 1-50SV (trade name, produced by Mitsubishi-Chemical Foods Corporation): PGFE (decaglycerol stearic acid ester)
Ryoto Polyglyester M-10D (trade name, Mitsubishi-Chemical Foods Corporation): PGFE (decaglycerol myristic acid ester)
NIKKOL HCO-60 (trade name, Nikko Chemicals Co., Ltd.): polyoxyethylene hydrogenated castor oil
NIKKOL TS-10V (trade name, Nikko Chemicals Co., Ltd.): polyoxyethylene sorbitan higher fatty acid ester (Polysorbate 60)
NIKKOL TO-10V (trade name, Nikko Chemicals Co., Ltd.): polyoxyethylene sorbitan higher fatty acid ester (Polysorbate 80)
NIKKOL TMGS-15V (trade name, Nikko Chemicals Co., Ltd.): polyoxyethylene glyceryl monostearate
X-ray diffraction measurement showed that the peak of crystalline curcumin disappeared completely or partially in Examples 1 to 9, Comparative Example 1, and Comparative Examples 4 to 7; and confirmed that the compositions obtained in these Examples and Comparative Examples contained amorphous curcumin. Differential scanning calorimetry showed that the peak of crystalline curcumin was reduced in Comparative Example 2, and the results confirmed that the composition obtained in Comparative Example 2 contained amorphous curcumin. Since the composition of Comparative Example 3 is merely a mixture of the components, the composition is assumed to contain crystalline curcumin.
A dissolution test was performed in accordance with the test method described in the Japanese Pharmacopoeia, 16th edition using the following materials under the following conditions. Analysis was performed by sampling a small amount of each test liquid at various times.
The test results are shown below.
Samples shown in Table 2 were used to test the changes over time of curcumin dissolution into artificial intestinal fluid, in comparison with a solid composition containing no surfactants, a solubilized preparation, and a solid composition prepared without heating. Table 3 and
Samples shown in Table 4 were used to test the changes over time of curcumin dissolution into artificial intestinal fluid, in comparison with other nonionic surfactants. The results are shown in Table 5 and
Samples shown in Table 6, which comprised various types of polyglycerol fatty acid esters, were used to test the changes over time of curcumin dissolution into artificial intestinal fluid. The results are shown in Table 7 and
Samples shown in Table 8, which comprised various amounts of polyglycerol fatty acid ester, were used to test the changes over time of curcumin dissolution into artificial intestinal fluid. The results are shown in Table 9 and
Samples shown in Table 10 were used to test the changes over time of curcumin dissolution into artificial intestinal fluid from compositions prepared using a sugar ester or lecithin, in comparison with the composition prepared using a polyglycerin fatty acid ester. The results are shown in Table 11 and
Compositions were prepared using HPC or HPMC in place of PVP in accordance with the production method described above, and subjected to the same tests as described above. The results showed that the compositions prepared using HPC or HPMC exhibited low ability to dissolve curcumin into body fluids (preferably intestinal fluid), as compared with the composition prepared using PVP; however, the same tendency as the composition using PVP was confirmed, and the dissolution of curcumin into body fluids (preferably intestinal fluid) was maintained for a prolonged period of time.
The changes over time of blood curcumin concentration in rats to which the amorphous preparation of Example 1 was administered was investigated by the following test method. As a comparative example, a bulk curcumin powder was administered.
Animals: Three SD rats (male, 7 weeks old, fasted for 14 to 16 hours before administration) per group were used.
Administration: 100 mg/kg (in terms of curcumin), single oral administration (sonde method)
Blood sampling: Jugular venous blood sampling immediately before administration; and 0.5, 1, 2, 4, 8, and 24 hours after administration
Analysis: 25 μl of plasma was enzymatically treated with β-glucuronidase. After extraction of curcumin with acetonitrile, the solvent was evaporated to dryness. The resulting product was rediluted with methanol, and measured by UV detection (420 nm).
In Test Example 8, the following preparations were used as samples.
Preparations 1 to 3 were produced in the same manner as the preparation of Example 1, except that the mixing ratio of the components was changed.
A cytotoxicity test was performed using the following samples under the following conditions, by the following methods.
Samples were diluted with PBS (phosphate-buffered physiological saline) to 3 mg/ml in terms of curcumin to prepare diluted samples.
Cells were seeded in a medium. After 24 hours of culture, the diluted samples were added in predetermined amounts. After 24 hours of culture, a WST8 reagent was added, and absorbance was measured.
Cells: B16F10 (skin cancer cells (metastatic cells)), HaCaT (human epidermal keratinocytes)
Medium: DMEM (high glucose), 10% FCS, 1% Ab
Culturing conditions: 37° C., 5% CO2, 24 hours
Samples: Preparation 1, Preparation 2, Preparation A, and Curcumin bulk powder.
Concentration of each sample added: 5, 10, and 20 μg/ml (in terms of curcumin).
As can be understood from these results, the preparation of the present invention showed a concentration-dependent efficacy in killing skin cancer cells in the tested concentration range, and acted more strongly on skin cancer cells than on normal cells.
This indicates that curcumin in the preparation of the present invention is readily absorbed by cells; and that the preparation of the present invention is effective for treating tumors, and has fewer side effects.
A cytotoxicity test was performed in the same manner as in Test Example 8-1, except that the following cells and samples were used.
This indicates that curcumin in the preparation of the present invention is readily absorbed by cells, and that the preparation of the present invention is effective for treating tumors.
A cytotoxicity test was performed in the same manner as in Test Example 8-1, except that the following cells and samples were used.
This indicates that curcumin in the preparation of the present invention is easily absorbed by cells, and that the preparation of the present invention is effective for treating tumors.
A cytotoxicity test was performed in the same manner as in Test Example 8-1, except that the following cells and samples were used.
This indicates that curcumin in the preparation of the present invention is readily absorbed by cells, and that the preparation of the present invention is effective for treating tumors.
A cytotoxicity test was performed using the following samples under the following conditions, by the following method.
A LDH-Cytotoxic Test Wako kit was used for this test. Samples were diluted with PBS to 3 mg/ml in terms of curcumin to prepare diluted samples.
Cells were seeded in a medium. After 24 hours of culture, the diluted samples were added in predetermined amounts.
After 24 hours of culture, a coloring reagent was added, and the cells were allowed to stand for 45 minutes. Absorbance (570 nm) was measured within 90 min after adding a reaction-stop solution.
Cell: B16F10 (skin cancer cells (metastatic cells))
Medium: DMEM (high glucose), 10% FCS, 1% Ab
Culture conditions: 37° C., 5% CO2, 24 hours
Preparation 3 and Preparation A
Concentration of sample added: 30 μg/ml (in terms of curcumin)
Coloring reagents: nitroblue tetrazolium, diaphorase, NAD
Reaction-stop reagent: hydrochloric acid (1 mol/L)
This indicates that curcumin in the preparation of the present invention is readily absorbed by cells, and that the preparation of the present invention is effective for treating tumors.
An insulin secretion test was performed using MIN6 cells (mouse pancreatic β cells) and the following samples under the following conditions, by the following method.
Samples were diluted with PBS to 3 mg/ml in terms of curcumin to prepare diluted samples.
Cells were seeded in a medium, and the diluted samples were added.
After 24 hours of culture, the cells were washed 3 times with KRBH buffer (0 mM glucose), and then incubated with KRBH buffer (0 mM glucose) for 1 hour.
Further, after washing once with KRBH buffer (0 mM glucose), the cells were incubated with KRBH buffer (25 mM glucose) for 24 hours.
The supernatant was collected, and insulin levels were measured by ELISA. (Absorbance measured at 450 nm.)
Cells: MIN6 cells (Mouse Pancreatic β Cells)
Media: DMEM (high glucose), 10% FCS, 1% Ab, 70 μM 2-ME
Culture conditions: 37° C., 5% CO2
Preparation 1
Concentration of the sample added: 1 μg/ml (in terms of curcumin)
As can be understood from the results, the preparation of the present invention increased insulin secretion in MIN6 cells (mouse pancreatic β-cells) by adding glucose.
The results show that curcumin in the preparation of the present invention is readily absorbed by cells, and that the preparation of the present invention is effective for treating diabetes.
Acute toxicity tests (organ weight measurement, biochemical testing, and blood cell testing) were performed using the following samples under the following conditions, by the following method.
The following samples were individually administered to the tail vein of mice (BALB/c). After 24 hours, the mice were dissected and subjected to organ weight measurement, biochemical testing, and blood cell testing. The biochemical testing was performed by measurement with a Fuji Dri-Chem, and the blood cell testing was performed by measurement with an XT-2000i multi-item automatic blood cell analyzer.
Amounts added: 0.25 mg/kg, 5 mg/kg, and 100 mg/kg (in terms of curcumin)
5 mg/kg is an amount that is almost equivalent to the ADI defined by WHO, and 100 mg/kg is about 30 times the ADI defined by WHO.
Control: containing all of the components of Preparation 3 other than curcumin
As can be understood from these results, even when curcumin was overdosed, the preparation of the present invention did not bring about any acute toxicity that would affect organ weight, biochemical markers, or blood cells.
A preparation comprising a CUR material, Kollidon K30, and PGFE (A) at a mixing ratio of 16:49:35 was produced by the same production method as for the preparation of Example 1.
A curcumin administration test was performed using HFD (high-fat diet)-challenged mice and the following samples under the following conditions, by the following methods.
In this test, total cholesterol (TCHO), chylomicron (CM), and LDL cholesterol (LDL) were measured.
Mice (C57BL/6, 5 weeks old, male, 6 to 7 mice per group) were allowed to freely drink an aqueous liquid of one of the samples described below, together with a high-fat diet (however, for the non-treated group described below, a normal diet) for 12 weeks, and evaluated for various parameters.
This test was performed using the test groups described below.
The following materials were given to the test groups.
Normal diet: D12450B (Research Diets)
High-fat diet: D12451 (Research Diets)
Bezafibrate: a 0.081 mass % aqueous Bezafibrate (Wako Pure Chemical Industries) liquid
Preparation A: a 0.04 mass % (in terms of curcumin) aqueous liquid of Preparation A (fine granules)
Example 1: a 0.04 mass % (in terms of curcumin) aqueous liquid of the preparation of Example 1
After the feeding period, blood was sampled and centrifuged at 3000G at 4° C. for 15 minutes to obtain plasma.
The plasma was measured by a Fuji Dri-Chem 4000V biochemical automatic analyzer (Fujifilm, Tokyo, Japan).
The results are shown in Tables 12 to 14, and
As can be understood from these results, it became clear that the preparation of Example 1 has an inhibitory effect on the elevation of THO, CM, and LDL.
The curcumin biodistribution after administration of curcumin preparations was evaluated under the following conditions, by the following methods.
Test samples were orally administered to rats (SD rats, 7 weeks old, male, fasted for 14 to 16 hours before administration, 3 mice per group), and the curcumin concentration in each organ was analyzed after 24 hours.
Dosage: 100 mg/kg in terms of curcumin
Administration method: single oral administration (sonde method)
Twenty-four hours after the administration, the rats were perfused with 50 ml of PBS or more to remove blood, and then dissected to remove organs.
Each organ was homogenized with 4 ml of 0.1% formic acid methanol per gram of the organ, and 500 μl of the homogenate was centrifuged at 10000G for 10 minutes to obtain a supernatant.
After the supernatant was evaporated to dryness under nitrogen and re-diluted with 80% methanol, the diluted supernatant was analyzed by UV detection (420 nm).
The results are shown in Tables 15 to 16, and
The tissue distribution after long-term ingestion of the preparation of the present invention was evaluated under the following conditions, by the following method.
Mice (BALB/c mice, 6 weeks old, male, 4 mice per group) were allowed to freely drink an aqueous liquid (concentration: 0.1% in terms of curcumin) of a curcumin preparation (preparation A or the preparation of Example 1) for 3 months, and the curcumin concentration in each organ was then analyzed.
After the mice were allowed to freely drink an aqueous solution of a curcumin preparation under the same conditions as above for 3 months, and then allowed to drink water in place of the aqueous solution for 24 hours, the curcumin concentration in each organ was analyzed.
After 3 months of the drinking, the mice were perfused with 5 ml of PBS or more to remove blood, and then dissected to remove organs.
Each organ was homogenized with 4 ml of 0.1% formic acid methanol per gram of the organ.
500 μl of each homogenate was centrifuged at 10000G for 10 minutes to obtain a supernatant.
After the supernatant was evaporated to dryness under nitrogen and re-diluted with 80% methanol, the diluted supernatant was measured by UV detection (420 nm).
The results are shown in Tables 17 to 20, and
A curcumin administration test was performed using HFD (high-fat diet)-challenged mice and the following samples under the following conditions, by the following method.
In this test, triglyceride (TG) was measured.
Mice (C57BL/6, 5 weeks old, male, 8 to 9 mice per group) were allowed to freely drink an aqueous liquid of the sample described below, together with a high-fat diet (however, for mice in the non-treated group described below, a normal diet) for 12 weeks, and evaluated for various parameters.
(1) Feed
Normal diet: D12450B (Research Diets)
High-fat diet: D12451 (Research Diets)
(2) Ingested Sample
Preparation B: Doctor's Best Curcumin Phytosome with Meriva (a commercially available curcumin preparation)
Example 10: 0.1 mass % (in terms of curcumin) aqueous liquid of Preparation A (fine granules) (test group)
After the feeding period, the blood was sampled and centrifuged at 3000G at 4° C. for 15 minutes to obtain plasma.
The plasma was measured by a Fuji Dri-Chem 4000V biochemical automatic analyzer (Fujifilm, Tokyo, Japan).
After the mice subjected to the test of Example 10 were perfused with 5 ml of PBS or more to remove blood, liver weight was measured.
Using an RNeasy Plus Mini Kit (Qiagen, Calif., USA), mRNA was extracted from the liver obtained by dissection. The mRNA was reverse-transcribed into cDNA using a High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific, Mass., USA).
Using the obtained cDNA as a template, a reaction liquid was prepared using a GeneAce SYBR qPCR Mix a Low ROX (Nippon Gene, Tokyo, Japan). The mRNA expression level of ACOX1 was determined by real-time PCR using a CFX384 (Bio-Rad Laboratories, CS, USA).
The liver weight increase and the ACOX1 expression level increase shown in the results suggest that the curcumin preparation of the present invention promoted PPAR-α activation.
The urinary curcumin level was evaluated by the following method.
Each curcumin preparation was orally, individually administered to rats at a dose of 100 mg/kg in terms of curcumin.
Urine was sampled over time, and the curcumin concentration in urine was analyzed.
a) 10 μL of a 0.1 M acetate buffer (pH 5.0) was added to the sampled urine.
For the measurement of the concentration of curcumin, inclusive of glucuronide conjugate, 25 μL of β-glucuronidase was then added, and the resulting mixture was mixed by a 10 s vortex and enzymatically treated.
b) Subsequently, 50 μL of a 200 ng/mL emodin solution was added.
c) Subsequently, 500 μL of an ethyl acetate/methanol=95/5 solution was added, and the resulting mixture was vortexed for 1 minute.
d) Subsequently, the resulting mixture was centrifuged at 10000G at 4° C. for 5 minutes, and the supernatant was collected in a 2-mL tube.
The procedures c) and d) were further repeated twice, and the collected supernatants were combined in a 2-mL tube.
The combined supernatants were evaporated to dryness under nitrogen.
On the day of analysis, 200 μL of 80% methanol was added to the nitrogen-dried sample, and the resulting mixture was vortexed for 1 minute.
The resulting mixture was centrifuged at 10000G at 4° C. for 5 minutes, and the supernatant was collected.
The supernatant was filtered through a 0.45 μm membrane filter, and analyzed by UV detection (wavelength 420 nm).
| Number | Date | Country | Kind |
|---|---|---|---|
| 2017-041173 | Mar 2017 | JP | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/JP2018/008186 | 3/2/2018 | WO | 00 |
