Berberrubine-related compound

Abstract

Provided are: a berberrubine-related compound having useful physiological activities; and a composition containing the berberrubine-related compound.　The composition contains quinoid-type berberrubine. Also provided are a berberrubine acetic acid adduct and an aqueous solution thereof. Also provided is a composition containing a berberrubine acetic acid adduct or an aqueous solution thereof. Also provided is a method for producing a berberrubine acetic acid adduct or an aqueous solution thereof. Also provided are a composition containing quinoid-type berberrubine and a composition containing a berberrubine acetic acid adduct or an aqueous solution thereof, each of which can be used as a pharmaceutical composition, a food composition, a cosmetic composition, or an agricultural composition.

Description

Berberrubin-related compounds

[0001] The present disclosure relates to berberrubine-related compounds, and in particular to a composition containing quinoid berberrubine, a berberrubine acetic acid adduct and a method for producing the same, and a composition containing the berberrubine acetic acid adduct.

[0002] Berberubine is a derivative of berberine, an alkaloid found in plants such as coptis and ambrosia. Berberine is a component of herbal medicines that has stomachic, anti-inflammatory, and tranquilizing effects, and berberubine is thought to be a metabolite of berberine in the body.

[0003] Non-Patent Document 1 describes that berberrubine can take the form of (i) a quinoid type, (ii) an enol type, or (iii) a zwitterionic type.

[0004] Spinozzi, S. et al., Journal of Natural Products, 77, 766-722, 2014

[0005] The present disclosure provides berberrubine-related compounds having useful physiological activities and compositions containing the same.

[0006] The present inventors have surprisingly found that compositions containing quinoid berberrubine have useful physiological activities, that a berberrubine acetic acid adduct can be obtained by adding acetic acid to quinoid berberrubine, and that compositions containing the berberrubine acetic acid adduct have high water solubility and useful physiological activities, and have thereby completed the present invention. Specifically, the present invention provides the following aspects.

[0007] [1] Formula (I): [2] A composition comprising a quinoid type berberrubine represented by the following formula (I):

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[0008] According to the present invention, it is possible to provide berberrubine-related compounds having useful physiological activities and compositions containing the same.

[0009] Figure 1A shows the solid state of quinoid berberrubine as determined by CPMAS. １３ Figure 1B shows the C NMR spectrum of quinoid berberrubine obtained by the DDMAS method. １３ 2A is a C NMR spectrum of the solid state of the berberrubine acetic acid adduct by CPMAS. １３ 2B is a C NMR spectrum of the solid state of the berberrubine acetic acid adduct obtained by the DDMAS method. １３ 3A is a C NMR spectrum of berberrubine acetate obtained by CPMAS. １３ 3B is a C NMR spectrum of berberrubine acetate obtained by the DDMAS method. １３Figure 4C NMR spectra. Figure 4A shows the time course of oxygen consumption rate (OCR) in C2C12 cells to which quinoid berberrubine was added. The time point 24 hours after addition was set as time 0 (min). An ATP synthase inhibitor (oligomycin), an uncoupler (carbonyl cyanide-m-chlorophenylhydrazone (CCCP)), and a mitochondrial complex inhibitor (rotenone and antimycin) were added at 20, 40, and 70 minutes (black arrows), respectively. Figure 4B shows the time course of oxygen consumption rate (OCR) in C2C12 cells to which berberrubine acetic acid adduct was added. The time point 24 hours after addition was set as time 0 (min). At 20, 40, and 70 minutes (black arrows), an ATP synthase inhibitor (oligomycin), an uncoupler (carbonyl cyanide-m-chlorophenylhydrazone (CCCP)), and mitochondrial complex inhibitors (rotenone and antimycin) were added, respectively. Figure 4C shows the OCR (corresponding to basal mitochondrial respiration, ATP production, and maximum mitochondrial respiration) at 30, 60, and 90 minutes (white arrows) in Figure 4A. Figure 4D shows the OCR (corresponding to basal mitochondrial respiration, ATP production, and maximum mitochondrial respiration) at 30, 60, and 90 minutes (white arrows) in Figure 4B. Figure 5 shows the results of detecting the formation of mitochondrial respiratory chain supercomplexes in C2C12 cells treated with quinoid berberrubine or berberrubine acetate adduct. Figure 6A shows a comparison of the expression levels of mitochondrial proteins OPA1, HSP60, and Mfn2 with the expression levels of cellular proteins Tub (α-tubulin) or Actin in primary cultured mouse fetal pituitary cells administered with a berberrubine acetic acid adduct. Figure 6B shows the results of primary cultured mouse fetal pituitary cells administered with a berberrubine acetic acid adduct, stained with Hoechst for nuclei and Tom20 for mitochondria, and observed under a fluorescence microscope (magnification: 60x). Figure 7 shows the results of Pink1 KO MEF cells and MEF cells administered with quinoid-type berberrubine or a berberrubine acetic acid adduct, stained with Hoechst for nuclei and Tom20 for mitochondria, and observed under a fluorescence microscope (magnification: 60x).A: Wild-type MEF cells (control, administered with DMSO only); B: Wild-type MEF cells administered with quinoid-type berberrubine; C: Wild-type MEF cells administered with the berberrubine acetic acid adduct. D: MEF_pink1 KO cells (control, administered with DMSO only); E: MEF_pink1 KO cells administered with quinoid-type berberrubine; F: MEF_pink1 KO cells administered with the berberrubine acetic acid adduct. Figure 8A shows the ratio of HeLa cell counts after administration of quinoid-type berberrubine (quinoid-type berberrubine) to the control HeLa cell counts (administered with DMSO only). Figure 8B shows the ratio of HeLa cell counts after administration of the berberrubine acetic acid adduct (berberrrubine acetic acid adduct) to the control HeLa cell counts (administered with water only). Figure 9 shows the ratio of MDS clone counts to blood cell counts in bone marrow transplant (BMT) mice administered berberrubine acetic acid adduct. Figure 10A shows the results of Western blotting analysis using antibodies against S396-phosphorylated tau (pS396), S422-phosphorylated tau (pS422), and full-length tau (JM) after adding quinoid berberrubine to Neuro2A cells containing intracellular aggregates of tau protein. Figure 10B shows the results of Western blotting analysis using antibodies against S396-phosphorylated tau (pS396), S422-phosphorylated tau (pS422), and full-length tau (JM) after adding berberrubine acetic acid adduct to Neuro2A cells containing intracellular aggregates of tau protein. Figure 10C shows the results of quantifying and comparing the tau protein detected in Figure 10A. Figure 10D shows the results of quantifying and comparing the tau protein detected in Figure 10B. Figure 11A shows snapshots of a video taken with a fluorescence microscope (magnification: 20x) of the motility of mouse sperm administered with the berberrubine acetic acid adduct or a control, after staining mitochondria. Figure 11B is a graph showing the number of sperm in the image of Figure 11A in which mitochondrial staining by MitoTracker was confirmed.Figure 11C is a snapshot of a video taken with an optical microscope (magnification: 20x) of mouse sperm administered with the berberrubine acetic acid adduct or a control. Figure 12 shows the effect of quinoid berberrubine on UV-irradiated mouse skin. + indicates a mouse administered with quinoid berberrubine, and - indicates a mouse not administered with quinoid berberrubine. Figure 13 shows the effect of quinoid berberrubine on the accumulation of mouse visceral fat. + indicates a mouse administered with quinoid berberrubine, and - indicates a mouse not administered with quinoid berberrubine. Figure 14 shows the results of Hoechst staining of nuclei and TMRM staining of mitochondria in brown adipocytes treated with or without quinoid berberrubine or the berberrubine acetic acid adduct, and observation of these cells under a fluorescence microscope (magnification: 60x). Figure 15A shows the growth of pea seedlings 3 days after administration of an aqueous solution of the berberrubine acetic acid adduct or the same amount of water. Figure 15B shows the growth of pea seedlings 9 days after administration of an aqueous solution of the berberrubine acetic acid adduct or an equal volume of water. Figure 15C shows the growth of pea seedlings 12 days after administration of an aqueous solution of the berberrubine acetic acid adduct or an equal volume of water. Figure 16 shows echocardiograms of mice administered with the berberrubine acetic acid adduct and control mice. Heart rate (HR), blood pressure (BP), left ventricular inflow / outflow ratio (E / A), heart rate, left ventricular end-diastolic diameter (LVDd), left ventricular end-systolic diameter (LVDs), left ventricular ejection fraction (LVEF), % left ventricular fractional shortening (%FS), and left ventricular filling pressure (E / e'). n = 7-9, error bars indicate standard deviation. Figure 17 shows heart weight, heart weight / body weight ratio, and lung weight / body weight ratio of mice administered with the berberrubine acetic acid adduct and control mice. n = 7-9, error bars indicate standard deviation. Figure 18 shows the expression of heart failure markers and myocardial fibrosis markers in mice administered with the berberrubine acetic acid adduct and control mice. n=7-9, error bars indicate standard deviation. Figure 19 shows the effect of the berberrubine acetic acid adduct on mitochondrial mRNA expression. n=6-9, error bars indicate standard deviation. Figure 20 shows echocardiograms of mice administered with the berberrubine acetic acid adduct or quinoid berberrubine and control mice.Heart rate, left ventricular end-diastolic dimension (LVDd), left ventricular end-systolic dimension (LVDs), left ventricular ejection fraction (LVEF), % left ventricular fractional shortening (%FS), and left ventricular filling pressure (E / e') are shown. n = 7-9, and error bars indicate standard deviation. * indicates p<0.05. Figure 21 shows the expression of heart failure markers and myocardial fibrosis markers in mice administered with berberrubine acetic acid adduct or quinoid-type berberrubine and control mice. n = 7-9, and error bars indicate standard deviation. *, **, and *** indicate p<0.05, p<0.01, and p<0.001, respectively. Figure 22 shows the effect on Mitol mRNA expression in mice administered with berberrubine acetic acid adduct or quinoid-type berberrubine. n = 7-9, and error bars indicate standard deviation. *, **, and *** indicate p<0.05, p<0.01, and p<0.001, respectively. ns indicates no significant difference. Figure 23A shows the time course of oxygen consumption rate (OCR) of mitochondria isolated from cardiomyocytes of control and berberrubine-treated mice treated with berberrubine acetic acid adduct or quinoid berberrubine. n=5, error bars indicate standard deviation. Measurements were started at 0 min, and ADP was added at 7 min, an enzyme inhibitor (oligomycin) at 15 min, a chemical uncoupler (carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP)) at 20 min, and mitochondrial complex inhibitors (rotenone and antimycin) at 25 min. The time period from 0 to 7 min was designated as state 2, from 7 to 15 min as state 3, from 15 to 20 min as state 4o, and from 20 to 25 min as state 3u. Figure 23B shows OCR in states 2, 3, 4o, and 3u in Figure 23A. Figure 24A shows the effects of the berberrubine acetate adduct on human dermal fibroblast (NHDF) cells and mitochondrial myopathy, encephalomyopathy, lactic acidosis, and stroke-like episodes syndrome (MELAS)-derived cells F954 (3243A>G, 92%) and F969 (3243A>G, 34.5%). The experimental start date was set as day 0, and water (control) or 10 μM berberrubine acetate adduct was administered on day 1, with oxygen consumption rate (OCR) measured on day 8.The measurement was started at 0 min, and an enzyme inhibitor (oligomycin) was added at 19 min, a chemical uncoupler (carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP)) at 45 min, and mitochondrial complex inhibitors (rotenone and antimycin) at 72 min. Figure 24B shows the basal respiration (the difference between OCR at 10 min and OCR at 90 min), maximum respiration (the difference between OCR at 63 min and OCR at 90 min), and spare respiratory capacity (the difference between OCR at 63 min and OCR at 10 min) from Figure 24A. Figure 25 shows the effect of berberrubine acetic acid adduct in a mouse inflammatory carcinogenesis model. Figure 26 shows the survival time and survival rate of mice administered with berberrubine acetic acid adduct and control mice.

[0010] [Explanation of Terms] Unless otherwise specified, each term in this disclosure has the following meaning.

[0011] In the present disclosure, "berberrubine" includes quinoid berberrubine, enol berberrubine, and zwitterionic berberrubine. Quinoid berberrubine has the IUPAC name 10-methoxy-5,6-dihydro-9H-[1,3]dioxolo[4,5-g]isoquinolino[3,2-a]isoquinolin-9-one and is represented by the following formula (1): The IUPAC name of the enol type berberrubine is 5,6-dihydro-9-hydroxy-10-methoxybenzo[g]-1,3-benzoxolo[5,6-a]quinolizinium, and is represented by the following formula (2): The IUPAC name of zwitterionic berberrubine is 10-methoxy-5,6-dihydro-[1,3]dioxolo[4,5-g]isoquinolino[3,2-a]isoquinolin-7-ium-9-olate, and is represented by the following formula (3): It is expressed as:

[0012] "Berberrubine-related compounds" include berberrubine, berberrubine derivatives, berberrubine precursors, as well as salts, acid adducts, and hydrates thereof.

[0013] In the present disclosure, a "berberrubine acetic acid adduct" is a compound in which acetic acid and / or acetate ions are added to berberrubine. Within the scope of this definition, the equivalent ratio of berberrubine to acetic acid and / or acetate ions is not limited. In the present disclosure, a berberrubine acetic acid adduct in which 1 equivalent of acetic acid and / or acetate ions is added to 1 equivalent of berberrubine is also referred to as a berberrubine acetate salt. When the equivalents of berberrubine and acetic acid and / or acetate ions for a certain berberrubine acetic acid adduct are determined based on experimental values, experimental error is taken into consideration when determining whether the berberrubine acetic acid adduct can be considered a berberrubine acetate salt. For example, when the equivalents of berberrubine and acetic acid and / or acetate ions ... １３ When determined by C NMR, a berberrubine acetic acid adduct determined to have one equivalent of acetic acid and / or acetate ion added to one equivalent of berberrubine can be considered to be a berberrubine acetate salt. Here, numerical values ​​can be interpreted, for example, by applying normal rounding, so that a berberrubine acetic acid adduct determined to have 0.5 to 1.4 equivalents of acetic acid and / or acetate ion added to one equivalent of berberrubine can be considered to be a berberrubine acetate salt, and a berberrubine acetic acid adduct determined to have 0.9 to 1.1 equivalents of acetic acid and / or acetate ion added to one equivalent of berberrubine can also be considered to be a berberrubine acetate salt.

[0014] The berberrubine acetic acid adduct used in the present disclosure is typically represented by the formula (i) (2): (ii) a berberrubine acetic acid adduct in which acetic acid and / or acetate ions are added to an enol-type berberrubine represented by formula (3): or (iii) a mixture thereof.

[0015] Unless otherwise specified, the term "berberine" used in this disclosure refers to berberine chloride. The IUPAC name for berberine chloride is 9,10-Dimethoxy-5,6-dihydro[1,3]dioxolo[4,5-g]isoquinolino[3,2-a]isoquinolin-7-ium chloride. Berberine chloride is commercially available (e.g., from Nacalai Tesque, Inc. or Tokyo Chemical Industry Co., Ltd. (TCI)). It can also be isolated from natural sources or artificially synthesized by known methods.

[0016] As used in this disclosure, the singular forms "a," "an," and "the" can include plural referents unless clearly indicated otherwise. As used in this disclosure, "and / or" includes both "and" and "or" relationships. "And" can also be expressed as "and" or "and." "Or" can also be expressed as "or." As used in this disclosure, "comprising" includes "consisting essentially of," "consisting essentially of," and "consisting," and "consisting essentially of" includes "consisting essentially of" and "consisting," and "consisting essentially of" includes "consisting essentially of."

[0017] Each numerical range in the present disclosure is intended to include the upper and lower limit values ​​indicated by "to" and "from." For example, the description "A to B" or "A to B" using numerical values ​​A and B means A or more and B or less. Furthermore, the descriptions "A to B," "A to B," or "A or more and B or less" in the numerical ranges described in stages in the present disclosure independently include both "A or more is preferred" and "B or less is preferred," and these lower or upper limit values ​​may be replaced with the upper or lower limit value of another numerical range. Furthermore, the lower or upper limit value of a numerical range described in the present disclosure is a numerical value within that numerical range and may be replaced with a numerical value shown in the examples.

[0018] Unless otherwise specified, all numbers expressing features, items, quantities, parameters, characteristics, periods, etc. used in the specification and claims are understood to be modified in all instances by the term "about." As used herein, the term "about" means that the so-specified feature, item, quantity, parameter, characteristic, or period encompasses a range above and below plus or minus 10 percent of the value of the stated feature, item, quantity, parameter, characteristic, or period. At least, and without limiting the application of the doctrine of equivalents to the scope of the claims, each numerical indicator should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Any numerical range or value inherently includes a range of error necessarily resulting from the standard deviation found in their respective testing measurements. Unless otherwise specified, each individual value of a numerical range herein is incorporated herein by reference as if it were individually recited herein.

[0019] [Embodiments] Hereinafter, embodiments of the present disclosure will be described in detail, but the scope of the present invention is not limited to the embodiments described herein, and various modifications can be made without departing from the spirit of the present invention. The embodiments may be implemented singly or in combination. For definitions and details of the embodiments, please refer to the "Explanation of Terms" above.

[0020] <Composition containing quinoid-type berberrubine> In one embodiment, a composition containing quinoid-type berberrubine is a compound of the following formula (I): A composition comprising a quinoid-type berberrubine represented by the formula:

[0021] The quinoid berberrubine may be isolated quinoid berberrubine, or may be a naturally occurring compound, an artificially synthesized compound, or a compound obtained by artificially modifying a naturally occurring raw material.

[0022] Quinoid berberrubine can be obtained, for example, by heating berberine chloride at 190°C for 3 hours under reduced pressure. Berberine chloride can be isolated from natural sources or artificially synthesized by known methods, and is also commercially available (e.g., from Nacalai Tesque, Inc. and Tokyo Chemical Industry Co., Ltd. (TCI)). Quinoid berberrubine is also commercially available (e.g., from Aurora Fine Chemicals and AMBINTER).

[0023] A composition containing quinoid berberrubine may contain quinoid berberrubine in any proportion. In certain embodiments, the content of quinoid berberrubine in a composition containing quinoid berberrubine may be 0.00001% by mass or more, 0.0001% by mass or more, 0.001% by mass or more, 0.01% by mass or more, 0.1% by mass or more, 1% by mass or more, 2% by mass or more, 5% by mass or more, 10% by mass or more, 20% by mass or more, 30% by mass or more, 40% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more, 80% by mass or more, 90% by mass or more, 95% by mass or more, 98% by mass or more, 99% by mass or more, or 100% by mass. Furthermore, a composition containing quinoid berberrubine may contain various additives within a range that does not impair the effects of the present invention. Non-limiting examples of additives include buffers, pH adjusters, surfactants, excipients, preservatives, antiseptics, and medicinal ingredients.

[0024] <Berberrubine Acetic Acid Adduct and Method for Producing the Same> In one embodiment, a berberrubine acetic acid adduct and a method for producing the same are provided.

[0025] Berberrubine acetic acid adduct can be produced by a production method comprising adding acetic acid to quinoid berberrubine. For example, 400 mL of acetic acid is added to quinoid berberrubine prepared from 50.0 g of berberine chloride, and the mixture is stirred at 40°C for 1 hour. 500 mL of diethyl ether is then added, and the mixture is stirred for an additional 3 hours. The mixture is then suction filtered, and the residue is dried under reduced pressure at room temperature to obtain the berberrubine acetic acid adduct as orange-yellow crystals.

[0026] Berberrubine acetic acid adduct is represented by the following formula (2): or an enol-type berberrubine represented by the following formula (3): The berberrubine acetic acid adduct may be a berberrubine acetic acid adduct in which acetic acid and / or acetate ions are added to a zwitterionic berberrubine represented by the following formula:

[0027] The berberrubine acetic acid adduct is a berberrubine acetic acid adduct in which one or more equivalents of acetic acid and / or acetate ions have been added to one equivalent of berberrubine. In one embodiment, the berberrubine acetic acid adduct is a berberrubine acetic acid adduct in which more than one equivalent, more than two equivalents, more than three equivalents, more than four equivalents, more than five equivalents, more than six equivalents, or more than seven equivalents of acetic acid and / or acetate ions have been added to one equivalent of berberrubine, or a berberrubine acetic acid adduct in which one or more equivalents, two or more equivalents, three or more equivalents, four or more equivalents, five or more equivalents, six or more equivalents, or seven or more equivalents of acetic acid and / or acetate ions have been added to one equivalent of berberrubine. For example, the berberrubine acetic acid adduct may be 2, 3, 4, 5, 6, 7, 8, or 9 equivalents in the range of 1 to 10 equivalents relative to one equivalent of berberrubine, or may be a range combining any of these values ​​as the upper and lower limits. For example, the berberrubine acetic acid adduct is a berberrubine acetic acid adduct to which 2 to 10 equivalents, 2 to 9 equivalents, 3 to 8 equivalents, or 4 to 7 equivalents of acetic acid and / or acetate ions have been added. Berberrubine acetic acid adducts to which acetic acid and / or acetate ions have been added in an amount within the above ranges per equivalent of berberrubine tend to be highly soluble in water.

[0028] Berberrubine acetic acid adduct: (i) a compound represented by the following formula (4): (ii) a compound represented by the following formula (5): The berberrubine acetic acid adduct may be a compound represented by formula (5), wherein n is 1 or more, or (iii) a mixture of compound (i) and compound (ii). n is not limited to an integer. In one embodiment, n is greater than 1, greater than 2, greater than 3, greater than 4, greater than 5, greater than 6, or greater than 7, or 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, or 7 or more. In another embodiment, n is in the range of 1 to 10, 2 to 9, 3 to 8, or 4 to 7. Berberrubine acetic acid adducts in which n is within these ranges tend to have high solubility in water.

[0029] The structure of the berberrubine acetic acid adduct is １３ This can be confirmed by C NMR. １３ As the C NMR method, for example, a crossed electrode magic angle spinning (CPMAS) method or a dipole decoupling magic angle spinning (DDMAS) method can be used. In addition, in the berberrubine acetic acid adduct, the equivalent of acetic acid and / or acetate ion added to 1 equivalent of berberrubine, and n in [Chemical Formula 4] and [Chemical Formula 5] are the solid １３ In C NMR analysis, the berberrubine equivalent can be calculated from the integral of the peak at the 8th carbon of berberrubine at 26 ppm or 27-28 ppm, and the acetic acid and / or acetate ion equivalent can be calculated from the integral of the peak at the CH carbon atom of acetic acid at 20-25 ppm, and then divided by the berberrubine equivalent. Alternatively, the berberrubine equivalent can be calculated from the integral of the peaks at the two carbon atoms of berberrubine at 55 ppm, and the acetic acid and / or acetate ion equivalent can be calculated from the integral of the peak at the C=O carbon atom of acetic acid at 180 ppm, and then divided by the berberrubine equivalent.

[0030] The solubility of the berberrubine acetic acid adduct in water may be 0.01 g / mL or more, 0.02 g / mL or more, 0.05 g / mL or more, 0.1 g / mL or more, 0.2 g / mL or more, 0.5 g / mL or more, or 1.0 g / mL or more. As described above, the solubility of the berberrubine acetic acid adduct in water can be adjusted by the amount of acetic acid and / or acetate ions added to berberrubine. Berberrubine acetic acid adducts in which one or more equivalents of acetic acid and / or acetate ions are added to one equivalent of berberrubine, or berberrubine acetic acid adducts in which n in [Chemical Formula 4] and [Chemical Formula 5] is 1 or more, tend to have high solubility in water. The solubility of the berberrubine acetic acid adduct (or other berberrubine-related compounds) in water or other solvents (e.g., methanol, dimethyl sulfoxide (DMSO), ethanol, or chloroform) can be determined by the following procedure, which is a partial modification of the method described in the General Rules of the Japanese Pharmacopoeia. A solvent is added to 1 g of a berberrubine-related compound, and the mixture is stirred for 30 seconds every 5 minutes at room temperature. After 30 minutes, it is confirmed whether the quinoid berberrubine and berberrubine-related compound have dissolved. The amount of solvent is appropriately adjusted to determine the minimum amount (mL) of solvent that can completely dissolve the berberrubine-related compound. The solubility (g / mL) of the berberrubine-related compound in each solvent is calculated from the berberrubine-related compound (1 g) and the minimum amount (mL) of solvent used. Berberrubine acetic acid adducts with a water solubility within the above-mentioned range tend to be easily administered or ingested by living organisms and also tend to be easily incorporated into pharmaceutical compositions, food compositions, agricultural compositions, etc.

[0031] <Aqueous Solution of Berberrubine Acetate Adduct> In one embodiment, an aqueous solution of berberrubine acetic acid adduct is provided. In the berberrubine acetic acid adduct, the added acetic acid and / or acetate ions are liberated in aqueous solution. Furthermore, the berberrubine acetic acid adduct is typically an acetic acid adduct of enol berberrubine and / or zwitterionic berberrubine. Therefore, the aqueous solution of berberrubine acetic acid adduct is an aqueous solution containing enol berberrubine and / or zwitterionic berberrubine.

[0032] The aqueous solution of the berberrubine acetic acid adduct may be an aqueous solution in which 0.0001 g or more, 0.001 g or more, 0.01 g or more, 0.05 g or more, or 0.1 g or more of the berberrubine acetic acid adduct is dissolved per mL of water, or 0.0001 to 1.0 g, 0.001 to 1.0 g, 0.01 to 1.0 g, or 0.05 to 1.0 g of the enol berberrubine and / or zwitterionic berberrubine per mL of water. The molar concentration of the enol berberrubine and / or zwitterionic berberrubine in this aqueous solution may be 0.0002 M or more, 0.002 M or more, 0.02 M or more, 0.1 M or more, or 0.2 M or more, or 0.0002 M to 2 M, 0.002 M to 2 M, 0.02 M to 2 M, or 0.1 M to 2 M.

[0033] The aqueous solution of the berberrubine acetic acid adduct may contain any other components as long as the effects of the present invention are not impaired. Non-limiting examples of other components include buffers, pH adjusters, surfactants, excipients, preservatives, antiseptics, and medicinal ingredients. Aqueous solutions containing berberrubine acetic acid adducts dissolved in these ranges are useful for administration to or ingestion by living organisms, and for incorporation into pharmaceutical compositions, food compositions, agricultural compositions, and the like.

[0034] <Method for Producing Berberrubine Acetic Acid Adduct and Its Aqueous Solution> In one embodiment, a method for producing a berberrubine acetic acid adduct or an aqueous solution thereof is provided. The berberrubine acetic acid adduct can be suitably produced by a production method comprising adding acetic acid to quinoid berberrubine. Here, acetic acid may be added directly to quinoid berberrubine powder or to a quinoid berberrubine solution.

[0035] In one embodiment, acetic acid is added as a liquid to quinoid berberrubine. For example, 400 mL of acetic acid is added to 40 g of quinoid berberrubine, and the mixture is stirred at 40° C. for 1 hour. Then, 500 mL of diethyl ether is added, and the mixture is stirred for an additional 3 hours. The mixture is then suction filtered, and the residue is dried at room temperature under reduced pressure (0.1 kPa) for 2 hours to obtain a berberrubine acetic acid adduct as orange-yellow crystals. Here, the berberrubine acetic acid adduct is typically a berberrubine acetic acid adduct in which more than one equivalent of acetic acid and / or acetate ions are added to one equivalent of berberrubine.

[0036] In another embodiment, acetic acid is added to quinoid berberrubine as acetic acid vapor. For example, a beaker containing quinoid berberrubine is left standing for one week in a sealed container saturated with acetic acid vapor while stirring with a spatula every other day, and then washed with diethyl ether and suction filtered to obtain the berberrubine acetic acid adduct as orange-yellow crystals. This berberrubine acetic acid adduct is typically a berberrubine acetic acid adduct in which more than one equivalent of acetic acid and / or acetate ions are added to one equivalent of berberrubine, and the amount of acetic acid and / or acetate ions added to berberrubine is approximately the same as that in the berberrubine acetic acid adduct prepared by the above-mentioned method in which acetic acid is not added as acetic acid vapor.

[0037] Berberrubine acetate salt (a berberrubine acetate adduct in which 1 equivalent of acetic acid and / or acetate ions are added to 1 equivalent of berberrubine) can be obtained as reddish-orange crystals by drying the berberrubine acetate adduct at a temperature higher than room temperature (e.g., 80°C) under reduced pressure for 5 hours. Furthermore, the amount of acetic acid and / or acetate ions added to berberrubine can be adjusted by appropriately adjusting the temperature and time.

[0038] Conventionally, the addition of acetic acid and / or acetate ions to a compound has been achieved by adding acetic acid to a solution of the compound in an organic solvent (e.g., diethyl ether or tetrahydrofuran) and precipitating the acetic acid adduct of the compound as an acetate salt insoluble in the organic solvent. This conventional method generally fails to produce acetic acid adducts of compounds with more than one equivalent of acetic acid and / or acetate ions added. On the other hand, the method of this embodiment differs from conventional methods in that it does not require dissolving quinoid berberrubine in an organic solvent. Furthermore, it is possible to prepare berberrubine acetic acid adducts with more than one equivalent of acetic acid and / or acetate ions added. Furthermore, it is also possible to adjust the amount of acetic acid and / or acetate ions added to berberrubine.

[0039] An aqueous solution of a berberrubine acetic acid adduct (including berberrubine acetate salt) can be prepared by dissolving the berberrubine acetic acid adduct (including berberrubine acetate salt) in water.

[0040] <Composition Comprising a Berberrubine Acetic Acid Adduct or an Aqueous Solution Thereof> In one embodiment, a composition containing a berberrubine acetic acid adduct or an aqueous solution thereof is provided. The composition containing a berberrubine acetic acid adduct or an aqueous solution thereof may contain the berberrubine acetic acid adduct in any proportion as long as the effects of the present invention are not impaired. In one embodiment, the content of the berberrubine acetic acid adduct may be 0.00001% by mass or more, 0.0001% by mass or more, 0.001% by mass or more, 0.01% by mass or more, 0.1% by mass or more, 1% by mass or more, 2% by mass or more, 5% by mass or more, 10% by mass or more, 20% by mass or more, 30% by mass or more, 40% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more, 80% by mass or more, 90% by mass or more, 95% by mass or more, 98% by mass or more, 99% by mass or more, or 100% by mass, based on the total mass of the composition containing the berberrubine acetic acid adduct. Furthermore, a composition containing a berberrubine acetic acid adduct or an aqueous solution thereof may contain any other components within the scope of not impairing the effects of the present invention. Non-limiting examples of other components include buffers, pH adjusters, surfactants, excipients, preservatives, antiseptics, and medicinal ingredients.

[0041] <Uses of the Composition> In one embodiment, a composition containing quinoid berberrubine and a composition containing a berberrubine acetic acid adduct or an aqueous solution thereof may be used for the same or different purposes. Furthermore, a composition containing quinoid berberrubine or a berberrubine acetic acid adduct or an aqueous solution thereof may be used for multiple purposes or for a single purpose.

[0042] A composition containing quinoid-type berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof can be administered to a subject in any form. For example, when the subject is a human or non-human animal, the composition of the present disclosure can be administered, for example, orally, intravenously, intramuscularly, intrathecally, sublingually, rectally, ophthalmically, otically, transauricularly, or transdermally. However, when the composition is a food composition, the subject can orally ingest it. When the subject is a plant, the composition can be administered to the plant by, for example, spraying or applying it to the plant or adding it to soil as fertilizer. The composition of the present disclosure may also be administered to a subject in vitro. In this case, the subject may be mitochondria, cells, or tissue. The mitochondria may be isolated from cells, tissue, or an individual; the cells may be isolated from tissue or an individual; and the tissue may be isolated from an individual. The cells may be primary cultured cells, subcultured cells, or cell lines. When administered to a subject in vitro, the compositions of the present disclosure can be administered to the subject by adding them directly to the subject or by adding them to the medium in which the subject is cultured.

[0043] (Pharmaceutical Composition) In one embodiment, a composition containing quinoid berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be a pharmaceutical composition. Pharmaceutical compositions include, for example, compositions contained in prescription drugs, over-the-counter drugs, or quasi-drugs. Furthermore, pharmaceutical compositions may be intended for use in humans or non-human animals.

[0044] The pharmaceutical composition is preferably a composition for activating mitochondria, treating or preventing a mitochondrial disease, treating or preventing cancer, treating or preventing a muscular disease, treating or preventing a neurological disease, treating or preventing a heart disease, improving or preventing cardiac hypertrophy, improving infertility, activating sperm activity, preventing the occurrence of wrinkles in the skin, reducing the depth and / or number of wrinkles, inhibiting an increase in the depth and / or number of wrinkles, inhibiting skin darkening, improving or preventing obesity, slowing the progression of aging, or extending lifespan.

[0045] A pharmaceutical composition may be formulated together with one or more other compositions, or may be formulated separately from one or more other compositions and used together.

[0046] "Treatment" includes the reduction, alleviation, or relief of disease symptoms, and "prevention" includes protection against future onset of a disease or symptom and the inhibition of progression. Desirable therapeutic effects of treatment include alleviation of symptoms, improvement of direct or indirect pathological consequences of a disease, reduction in the rate of progression of worsening symptoms, recovery or alleviation of the disease state, and improvement in prognosis. For example, treatment or prevention of cancer includes administering to a subject with cancer or diagnosed with cancer a pharmaceutical composition containing quinoid berberrubine or berberrubine acetic acid adduct, or an aqueous solution thereof, for the purpose of achieving at least one desired therapeutic effect, such as a reduction in the number of cancer cells, a reduction in tumor size, a reduction in the rate of cancer cell invasion into peripheral organs, or a reduction in tumor metastasis or tumor growth rate.

[0047] Pharmaceutical compositions can be in the form of, for example, tablets, granules, powders, capsules, emulsions, suspensions, syrups, or injections such as sterile solutions and suspensions. When two or more pharmaceutical compositions are formulated separately to form two or more formulations, the individual formulations can be administered simultaneously, separately at a certain time interval, or consecutively. The two or more formulations can also be administered at different times per day. Furthermore, pharmaceutical compositions can be administered systemically or locally, orally or parenterally. When active ingredients are formulated separately to form two or more formulations, the individual formulations can be administered by different routes.

[0048] (Food Composition) In one embodiment, a composition containing quinoid berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be a food composition. Food compositions include, in addition to general foods and beverages, compositions contained in, for example, nutritional foods and beverages, specialty foods and beverages, nutritional supplements and nutritional supplement beverages (supplements), and food additives. Furthermore, food compositions may be intended for use by humans or non-human animals.

[0049] The food composition is preferably a composition for activating mitochondria, improving or preventing symptoms associated with mitochondrial diseases, improving or preventing symptoms associated with cancer, improving or preventing symptoms associated with muscle diseases, improving or preventing symptoms associated with neurological diseases, improving or preventing symptoms associated with heart diseases, improving or preventing cardiac hypertrophy, improving symptoms associated with infertility, stimulating sperm activity, preventing the occurrence of wrinkles in the skin, reducing the depth and / or number of wrinkles, inhibiting an increase in the depth and / or number of wrinkles, inhibiting skin darkening, improving or preventing obesity, slowing the progression of aging, or extending lifespan.

[0050] The food composition may be formulated together with one or more other compositions, or may be formulated separately from one or more other compositions and used in combination.

[0051] The food composition can be in the form of a beverage, solid, liquid, paste, powder, etc. For example, it can be a beverage, soup, processed meat product, processed vegetable product, processed fruit product, paste-like seasoning, concentrated paste food, granular seasoning, concentrated granular food, supplement, etc. Here, processed products refer to natural food ingredients that have been processed and / or cooked, and include frozen foods, retort foods, canned foods, bottled foods, etc. Supplements (nutritional supplements, nutritional supplement drinks) can be in the form of, for example, liquid drinks, capsules, tablets, powders, etc. The food composition of the present disclosure, or a food containing the food composition, may be included in a food product that claims to have the following effects: mitochondrial activation, improvement or prevention of symptoms associated with mitochondrial disease, improvement or prevention of symptoms associated with cancer, improvement or prevention of symptoms associated with muscle disease, improvement or prevention of symptoms associated with neurological disease, improvement or prevention of symptoms associated with heart disease, improvement or prevention of cardiac hypertrophy, improvement of symptoms associated with infertility, activation of sperm activity, prevention of the occurrence of wrinkles in the skin, reduction of wrinkle depth and / or number, inhibition of increase in wrinkle depth and / or number, inhibition of skin darkening, improvement or prevention of obesity, inhibition of the aging process, or extension of lifespan. The food composition or food product may be provided in a sealed form in a bag, container, or the like. The bag and container used in the present disclosure may be any bag and container commonly used for food.

[0052] (Cosmetic Composition) In one embodiment, a composition containing quinoid berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be a cosmetic composition. Cosmetic compositions include, for example, compositions intended for the beautification of skin, hair, and nails. Furthermore, the cosmetic composition may be intended for use by humans or non-human animals.

[0053] Preferably, the cosmetic composition is a composition for activating mitochondria, for preventing the occurrence of wrinkles in the skin, for reducing the depth and / or number of wrinkles, for inhibiting the increase in the depth and / or number of wrinkles, for inhibiting darkening of the skin, or for slowing the progression of aging.

[0054] The cosmetic composition may be formulated together with one or more other compositions, or may be formulated separately from one or more other compositions and used in combination.

[0055] Cosmetic compositions are compositions intended for the purpose of beautifying, for example, skin, hair, nails, etc., and can be formulated in a form that allows for the care of these areas. For example, they may be liquids, lotions, creams, patches, oils, sprays, liquid cleansers, solid soaps, etc. Furthermore, products may also include cosmetic liquids, beauty serums, moisturizing liquids, moisturizing creams, soaps, body soaps, bath salts, sunscreens, shaving lotions, depilatories, shampoos, conditioners, hair tonics, hair dyes, etc.

[0056] (Agricultural Compositions) In one embodiment, a composition containing quinoid berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be an agricultural composition. Agricultural compositions include, for example, compositions for activating mitochondria, compositions containing nutrients or nutritional supplements for agricultural crops, etc., compositions for promoting or suppressing the growth of agricultural crops, etc., compositions used to enhance or suppress the physiological functions of agricultural crops, etc., and compositions for treating or preventing diseases of agricultural crops, etc. Agricultural crops, etc. include trees, bamboo, ornamental plants, aquatic plants, and agricultural and forestry products.

[0057] The agricultural composition is preferably a composition for promoting the growth of branches and leaves, the growth of flowers and fruits, and the establishment of roots, for delaying the progression of aging, or for extending the lifespan of plants such as agricultural crops.

[0058] The agricultural composition may be formulated together with one or more other compositions, or may be formulated separately from one or more other compositions and used in combination.

[0059] The agricultural composition can be formulated as a solid such as a powder, granule, encapsulated granule, fine granule, microgranule capsule, microcapsule, pellet, dispersible concentrate, tablet, flake, or as a liquid in which the active ingredient is dissolved in a liquid component. The product may be a pesticide, fertilizer, fertilized potting soil, etc.

[0060] (Composition for Activating Mitochondria) In one embodiment, a composition containing quinoid-type berberrubine or a berberrubine acetic acid adduct or an aqueous solution thereof may be a composition for activating mitochondria. Activating mitochondria may mean improving mitochondrial function (e.g., respiratory capacity) or suppressing a decline in mitochondrial function. Mitochondria are intracellular organelles of eukaryotes and perform important functions in the healthy life activities of eukaryotes, such as efficiently producing ATP through aerobic respiration. By effectively activating mitochondria, the composition of the present disclosure can maintain individuals, tissues, and cells in a healthy state and can prevent, ameliorate, or treat mitochondrial diseases, as described below, and various diseases and symptoms caused by at least one of the onset factors of a decline in mitochondrial function. Furthermore, the composition of the present disclosure can be preferably used as a pharmaceutical composition for activating mitochondria, a food composition, a cosmetic composition, or an agricultural composition.

[0061] (Composition for Mitochondrial Disease) In one embodiment, a composition containing quinoid berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be a composition for treating or preventing mitochondrial disease. Mitochondrial disease is a disease caused by abnormal mitochondrial function. Within this definition, mitochondrial disease includes any pathological condition, but non-limiting examples include, for example, neurological diseases, muscular diseases, cardiac diseases, or infertility, which are described below and in which impaired mitochondrial function is at least one of the causes of the onset. For example, the mitochondrial disease may be Parkinson's disease or mitochondrial myopathy, encephalomyopathy, lactic acidosis, and stroke-like episodes syndrome (MELAS). MELAS is a type of mitochondrial disease characterized by recurrent stroke-like episodes and is typically caused by a point mutation (3243A>G mutation) in mtDNA. The composition of the present disclosure can be preferably used as a pharmaceutical composition or a food composition for treating or preventing mitochondrial disease.

[0062] (Composition for Neurological Disease) In one embodiment, a composition containing quinoid berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be a composition for treating or preventing a neurological disease. A neurological disease is a disease in which neurological damage occurs due to a lesion in the nerve. Within this definition, neurological diseases include any pathological condition, but non-limiting examples include Parkinson's disease, dementia, Guillain-Barré syndrome, Pick's disease, and Huntington's disease. Non-limiting examples of dementia include Alzheimer's disease. Neurological diseases may be caused by at least one cause of mitochondrial dysfunction, but are not limited to this.

[0063] The treatment or prevention of a neurological disease may be the improvement or prevention of at least one clinical symptom observed in the neurological disease. The treatment or prevention of Parkinson's disease may be the improvement or prevention of one or more symptoms observed in Parkinson's disease, such as tremor (tremors), bradykinesia, rigidity (muscle stiffness), and impaired posture (proneness to falling). The treatment or prevention of dementia may be the improvement or prevention of one or more symptoms observed in dementia, such as memory impairment, aphasia, and executive dysfunction. In some embodiments, the treatment or prevention of dementia may be the suppression of a pathological condition seen in dementia, such as the accumulation of tau protein in brain neurons, or the removal of at least a portion of tau protein accumulated in brain neurons.

[0064] Furthermore, the treatment or prevention of a neurological disease may involve suppressing or at least partially removing the accumulation of tau protein, or suppressing the phosphorylation of S396 and / or S422 of tau protein. Furthermore, the treatment or prevention of a neurological disease may involve inhibiting the activity of GSK3β or JNK. The composition of the present disclosure can be preferably used as a pharmaceutical composition or a food composition for the treatment or prevention of a neurological disease.

[0065] (Composition for Muscle Disease) In one embodiment, a composition containing quinoid berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be a composition for treating or preventing a muscle disease.

[0066] Muscle diseases are disorders that cause damage due to muscle lesions. Muscles include, for example, skeletal muscles, visceral muscles, smooth muscles, and cardiac muscles. Within this definition, muscle diseases include any pathological condition, but non-limiting examples include Parkinson's disease, frailty, locomotive syndrome, sarcopenia, amyotrophic lateral sclerosis, myasthenia gravis, muscular dystrophy, polymyositis, cardiomyopathy (cardiomyopathy disease accompanied by cardiac dysfunction), and mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episode syndrome (MELAS). Muscle diseases may be caused by, but are not limited to, reduced mitochondrial function. Note that muscle diseases in the present disclosure also include concepts aimed at promoting health, such as frailty, locomotive syndrome, and sarcopenia, and concepts aimed at preventing progression to more severe physical conditions.

[0067] Frailty is a condition characterized by various changes in organ function and a decline in organ reserve with aging, leading to increased vulnerability to external stress. Sarcopenia is a syndrome characterized by a progressive and systemic loss of muscle mass and strength, and is associated with physical dysfunction, reduced quality of life, and the risk of death. Locomotive syndrome is a condition in which musculoskeletal disorders limit daily life, leading to the need for care or assistance or increasing the risk of such. Sarcopenia is a concept encompassed within locomotive syndrome, and both sarcopenia and locomotive syndrome are encompassed within frailty. Mitochondrial dysfunction has been recognized as one of the mechanisms underlying the onset and progression of sarcopenia. The compositions of the present disclosure can be preferably used as pharmaceutical compositions or food compositions for the treatment or prevention of frailty, locomotive syndrome, and sarcopenia.

[0068] (Composition for Cancer) In one embodiment, a composition containing quinoid-type berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be a composition for treating or preventing cancer. Cancer refers to any cancer, including solid cancers and blood cancers, and non-limiting examples include epithelial cancer, carcinoma, cervical cancer, sarcoma, and myelodysplastic syndrome (MDS). Non-limiting examples of myoblastoma include fibrosarcoma, odontogenic sarcoma, malignant fibrous histiocytoma, dermatofibrosarcoma, liposarcoma, rhabdomyosarcoma, leiomyosarcoma, angiosarcoma, Kaposi's sarcoma, lymphangiosarcoma, synovial sarcoma, alveolar soft part sarcoma, extraskeletal chondrosarcoma, and extraskeletal osteosarcoma. The cancer may also be an inflammatory cancer. Inflammatory cancer may be a cancer accompanied by inflammation or may be a cancer caused by inflammation. Inflammation includes chronic inflammation and acute inflammation. Inflammatory cancer includes malignant cancers such as metastatic and / or invasive cancers. Inflammatory cancer may be, for example, cancer caused by inflammation-associated carcinogenesis. The composition of the present disclosure can be preferably used as a pharmaceutical composition or a food composition for treating or preventing cancer.

[0069] (Composition for Cardiac Disease / Cardiac Hypertrophy) In one embodiment, a composition containing quinoid-type berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be a composition for treating or preventing cardiac disease and / or a composition for ameliorating or preventing cardiac hypertrophy. Cardiac disease includes any heart-related disease or symptom within the scope of the effects of the present invention, including, but not limited to, heart failure, cardiomyopathy, valvular disease, atrial septal defect, cardiac tumor, arrhythmia, cardiac hypertrophy, ischemic heart disease, etc. Heart failure refers to various systemic disorders resulting from impaired cardiac function, and its presence and progression can be effectively evaluated using heart failure markers, as described below. Cardiomyopathy refers to a myocardial disease accompanied by cardiac dysfunction, and its presence and progression can be effectively evaluated using myocardial fibrosis markers, as described below. Cardiac hypertrophy refers to a condition in which the myocardium and / or ventricular lumen become enlarged, which tends to impair cardiac function and / or increase the risk of the aforementioned heart diseases. Furthermore, the heart disease may include cardiac hypertrophy as at least one of its symptoms, or may have cardiac hypertrophy as at least one of its causes.

[0070] The composition of the present disclosure may reduce the expression of a heart failure marker in an individual, tissue, or cell. In this case, the composition of the present disclosure may reduce the expression of the heart failure marker in the individual, tissue, or cell by preferably 5% or more, more preferably 10% or more, compared to the expression of the heart failure marker in an individual, tissue, or cell to which the composition of the present disclosure is not administered. Any heart failure marker may be used as long as the effects of the present invention are achieved. Non-limiting examples include one or more mRNAs of Nppa, Nppb, and Myh7. Any myocardial fibrosis marker may be used as long as the effects of the present invention are achieved. Non-limiting examples include one or more mRNAs of Collagen 1 and Ctgf.

[0071] The composition of the present disclosure may reduce the expression of myocardial fibrosis markers in an individual, tissue, or cell. In this case, the composition of the present disclosure may reduce the expression of myocardial fibrosis markers in an individual, tissue, or cell by preferably 5% or more, more preferably 10% or more, compared to the expression of myocardial fibrosis markers in an individual, tissue, or cell to which the composition of the present disclosure is not administered. The composition of the present disclosure can be preferably used as a pharmaceutical composition or a food composition for the treatment or prevention of heart disease.

[0072] (Composition for Infertility) In one embodiment, a composition containing quinoid berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be a composition for improving infertility. The composition for improving infertility may be a composition for improving male infertility, and the composition for improving male infertility may be a composition for activating sperm activity. At least one of the causes of infertility may be, but is not limited to, a decrease in mitochondrial function. The composition of the present disclosure can be preferably used as a pharmaceutical composition or a food composition for improving infertility.

[0073] (Cosmetic Composition) In one embodiment, a composition containing quinoid berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be a composition intended for cosmetic purposes on areas such as skin, hair, and nails, and may preferably be a composition for improving skin conditions or suppressing or preventing deterioration of the condition.

[0074] The skin conditions include wrinkles, darkening (age spots), warts, sagging, dryness, etc., and their deterioration means, for example, with respect to wrinkles, the occurrence of wrinkles, an increase in the depth and / or number of wrinkles; with respect to darkening, the occurrence of darkening, a darkened area becoming darker and / or an enlargement of the darkened area; with respect to warts, the occurrence of warts, an increase in the size and / or number of warts; with respect to sagging, the occurrence of sagging, an increase in the size and / or number of sagging; with respect to dryness, the occurrence of dryness and / or an enlargement of the dry area.

[0075] For example, a composition containing quinoid berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be a composition for preventing the occurrence of wrinkles in the skin, a composition for reducing the depth and / or number of wrinkles, and / or a composition for inhibiting an increase in the depth and / or number of wrinkles, or a composition for inhibiting skin darkening (the occurrence of darkening, the darkening of darkened areas, and / or the expansion of darkened areas).

[0076] The target skin is human or non-human animal skin, preferably human mammal skin. The target skin area includes the hands, face (forehead, cheeks, eye contours (around the eyeballs), corners of the eyes, under the eyes, eyelids, etc.), neck, legs, feet, arms, elbows, abdomen, back, etc. The wrinkles or darkening are not limited by their cause and may be caused by ultraviolet exposure, aging, side effects of medication, etc.

[0077] The composition of the present disclosure can be preferably used as a pharmaceutical composition, a food composition, or a cosmetic composition for cosmetic purposes.

[0078] (Composition for ameliorating or preventing obesity) In one embodiment, a composition containing quinoid-type berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be a composition for ameliorating or preventing obesity. A composition for ameliorating or preventing obesity may be a composition for reducing or preventing the accumulation of body fat. The body fat may be visceral fat or subcutaneous fat, i.e., the obesity may be visceral fat obesity or subcutaneous fat obesity. The reduction in body fat accumulation may be due to, for example, a reduction in white adipocytes. The reduction in white adipocytes is preferably due to the activation of brown adipocytes. The activation of brown adipocytes is preferably the activation of mitochondrial function in brown adipocytes. The composition of the present disclosure can be preferably used as a pharmaceutical composition or a food composition for the amelioration or prevention of obesity.

[0079] (Composition for Delaying Aging) In one embodiment, a composition containing quinoid berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be a composition for delaying the progression of aging. Aging refers to the progression of changes in physical characteristics, declines in tissue and cell function, and the like that accompany aging. Non-limiting examples of aging include an increased risk of age-related diseases, muscle weakness, skin aging (increased depth and / or number of wrinkles, increased age spots, warts, sagging, etc.), and hair aging (graying, loss of volume, and deterioration of hair quality). Non-limiting examples of diseases that increase the risk of disease with aging include mitochondrial diseases, cancer, muscle diseases, neurological diseases, and heart diseases. The composition of the present disclosure can be preferably used as a pharmaceutical composition, food composition, or cosmetic composition for delaying the progression of aging.

[0080] (Composition for extending lifespan) In one embodiment, a composition containing quinoid berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof may be a composition for extending lifespan or survival time. Here, lifespan may be the entire survival period from birth to death of an individual, tissue, and / or cell that is not due to external causes of death (e.g., a traffic accident). Survival time may be the entire survival period from birth to death of a tissue and / or cell, or may be the survival period from the start of administration of a composition containing quinoid berberrubine or berberrubine acetic acid adduct or an aqueous solution thereof to death.

[0081] The composition of the present disclosure can extend the lifespan or survival period of an individual, tissue, and / or cell by preferably 5 weeks or more, more preferably 10 weeks or more, and particularly preferably 20 weeks or more, compared to the lifespan or survival period of an individual, tissue, and / or cell to which the composition of the present disclosure is not administered. Furthermore, the composition of the present disclosure can extend the lifespan or survival period of a human to which the composition of the present disclosure is not administered by preferably 150 weeks or more, more preferably 300 weeks or more, and particularly preferably 600 weeks or more. Furthermore, the composition of the present disclosure can extend the lifespan or survival period of an individual, tissue, and / or cell to which the composition of the present disclosure is not administered by preferably 1.05 times or more, more preferably 1.1 times or more, and particularly preferably 1.2 times or more.

[0082] Furthermore, the composition of the present disclosure can extend the 50% survival time of an individual, tissue, and / or cell by preferably 5 weeks or more, more preferably 10 weeks or more, and particularly preferably 20 weeks or more, compared to the 50% survival time of an individual, tissue, and / or cell not administered with the composition of the present disclosure. The composition of the present disclosure can extend the 50% survival time of a human by preferably 150 weeks or more, more preferably 300 weeks or more, and particularly preferably 600 weeks or more, compared to the 50% survival time of a human not administered with the composition of the present disclosure. The composition of the present disclosure can extend the 50% survival time of an individual, tissue, and / or cell by preferably 1.05 times or more, more preferably 1.1 times or more, and particularly preferably 1.2 times or more, compared to the 50% survival time of an individual, tissue, and / or cell not administered with the composition of the present disclosure. The 50% survival time may be the 50% survival time from the birth of the individual, tissue, and / or cell, or may be the 50% survival time from the start of administration of the composition of the present disclosure.

[0083] The composition of the present disclosure can be preferably used as a pharmaceutical composition for extending lifespan or as a food composition.

[0084] (Composition for Promoting Plant Growth) In one embodiment, a composition containing quinoid berberrubine or a berberrubine acetic acid adduct or an aqueous solution thereof may be a composition for promoting plant growth. The target plant may be either a seed plant or a non-seed plant, but is preferably a seed plant. The seed plant may be either an angiosperm or a gymnosperm. Gymnosperms may be plants classified in the Cycadidae, Ginkgoidea, Pinidae, Taxidae, Gnetophyta, etc., and angiosperms may be plants classified in the Monocotyledonous, Dicotyledonous, etc. The promotion of plant growth may be one or more effects related to growth, such as promoting the growth of branches and leaves, flower and fruit growth, and / or root development, of plants such as agricultural crops. The composition of the present disclosure can be preferably used as an agricultural composition for promoting plant growth.

[0085] Further embodiments of the present disclosure are exemplified below. [1] A compound of the following formula (I): A composition comprising a quinoid berberrubine represented by the formula (I). [2] The composition according to [1], wherein the compound represented by formula (I) is isolated quinoid berberrubine. [3] The composition according to [1] or [2], which is a pharmaceutical composition. [4] The composition according to any of [1] to [3], which is a food composition. [5] The composition according to any of [1] to [4], which is a cosmetic composition. [6] The composition according to any of [1] to [5], which is an agricultural composition. [7] The composition according to any of [1] to [6], which is a composition for activating mitochondria. [8] The composition according to any of [1] to [7], which is a composition for treating or preventing mitochondrial disease. [9] The composition according to any of [1] to [8], which is a composition for treating or preventing Parkinson's disease.

[10] The composition according to any of [1] to [9], which is a composition for treating or preventing mitochondrial myopathy, encephalomyopathy, lactic acidosis, and stroke-like episodes syndrome (MELAS).

[11] The composition according to any of [1] to

[10] , which is a composition for treating or preventing cancer.

[12] The composition according to

[11] , wherein the cancer is epithelial cancer, carcinoma, cervical cancer, sarcoma, or myelodysplastic syndrome (MDS).

[13] The composition according to any one of [1] to

[12] , wherein the composition is for treating or preventing a muscular disease.

[14] The composition according to

[13] , wherein the muscular disease is frailty, locomotive syndrome, or sarcopenia.

[15] The composition according to any one of [1] to

[14] , wherein the composition is for treating or preventing a neurological disease.

[16] The composition according to

[15] , wherein the neurological disease is dementia.

[17] The composition according to any one of [1] to

[16] , wherein the composition is for treating or preventing a cardiac disease.

[18] The composition according to

[17] , wherein the cardiac disease is heart failure.

[19] The composition according to

[17] or

[18] , wherein the cardiac disease is cardiomyopathy.

[20] The composition according to any one of [1] to

[20] , wherein the composition is for ameliorating or preventing cardiac hypertrophy.

[21] The composition according to any one of [1] to

[20] , which is a composition for improving infertility.

[22] The composition according to any one of [1] to

[21] , which is a composition for activating sperm activity.

[23] The composition according to any one of [1] to

[22] , which is a composition for preventing the occurrence of wrinkles on the skin, reducing the depth and / or number of wrinkles, and / or inhibiting an increase in the depth and / or number of wrinkles.

[24] The composition according to any one of [1] to

[23] , which is a composition for inhibiting or preventing skin darkening.

[25] The composition according to any one of [1] to

[24] , which is a composition for improving or preventing obesity.

[26] The composition according to any one of [1] to

[25] , which is a composition for slowing the progression of aging.

[27] The composition according to any one of [1] to

[26] , which is a composition for extending lifespan.

[28] The composition according to any one of [1] to

[27] , which is a composition for promoting plant growth.

[29] A compound represented by the following formula (I):

[30] A method for producing a berberrubine acetic acid adduct, comprising adding acetic acid to a quinoid type berberrubine represented by the formula (I):

[31] (i) A method for producing a berberrubine acetic acid adduct, comprising adding acetic acid as acetic acid vapor, according to

[29] . (ii) adding acetic acid to a quinoid-type berberrubine represented by the formula (2): to generate a berberrubine-acetic acid adduct; and (ii) dissolving the generated berberrubine-acetic acid adduct in water.

[32] The method according to

[31] , wherein in (i), acetic acid is added as acetic acid vapor to generate the berberrubine-acetic acid adduct.

[33] Berberrubine-acetic acid adduct.

[34] A berberrubine-acetic acid adduct of the following formula (2): or an enol-type berberrubine represented by the following formula (3):

[35] The berberrubine acetic acid adduct according to

[33] or

[34] , in which acetic acid and / or acetate ions are added to a zwitterionic berberrubine represented by the following formula (4):

[36] (i) The berberrubine acetic acid adduct according to

[33] or

[34] , in which 1 equivalent or more of acetic acid and / or acetate ions are added to 1 equivalent of berberrubine: (ii) a compound represented by the following formula (5): [In formula (5), n is 1 or more], or (iii) a mixture of compound (i) and compound (ii).

[37] The berberrubine acetic acid adduct according to any of

[33] to

[36] , which has a solubility in water of 0.01 g / mL or more.

[38] An aqueous solution of the berberrubine acetic acid adduct according to any of

[33] to

[37] .

[39] A composition comprising the berberrubine acetic acid adduct according to any of

[33] to

[37] or the aqueous solution according to

[38] .

[40] The composition according to

[39] , which is a pharmaceutical composition.

[41] The composition according to

[39] or

[40] , which is a food composition.

[42] The composition according to any of

[39] to

[41] , which is a cosmetic composition.

[43] The composition according to any of

[39] to

[42] , which is an agricultural composition.

[44] The composition according to any of

[39] to

[43] , which is a composition for activating mitochondria.

[45] The composition according to any one of

[39] to

[44] , which is a composition for treating or preventing a mitochondrial disease.

[46] The composition according to any one of

[39] to

[42] , which is a composition for treating or preventing Parkinson's disease.

[47] The composition according to any one of

[39] to

[46] , which is a composition for treating or preventing mitochondrial myopathy, encephalomyopathy, lactic acidosis, and stroke-like episodes syndrome (MELAS).

[48] The composition according to any one of

[39] to

[47] , which is a composition for treating or preventing cancer.

[49] The composition according to

[48] , wherein the cancer is epithelial cancer, carcinoma, cervical cancer, sarcoma, or myelodysplastic syndrome (MDS).

[50] The composition according to any one of

[39] to

[49] , which is a composition for treating or preventing a muscular disease.

[51] The composition according to

[50] , wherein the muscular disease is frailty, locomotive syndrome, or sarcopenia.

[52] The composition according to any one of

[39] to

[51] , which is a composition for treating or preventing a neurological disease.

[53] The composition according to

[52] , wherein the neurological disease is dementia.

[54] The composition according to any one of

[39] to

[53] , which is a composition for treating or preventing a cardiac disease.

[55] The composition according to

[54] , wherein the cardiac disease is heart failure.

[56] The composition according to

[54] or

[55] , wherein the heart disease is cardiomyopathy.

[57] The composition according to any of

[39] to

[56] , wherein the composition is for improving or preventing cardiac hypertrophy.

[58] The composition according to any of

[39] to

[57] , wherein the composition is for improving infertility.

[59] The composition according to any of

[39] to

[58] , wherein the composition is for activating sperm activity.

[60] The composition according to any of

[39] to

[59] , wherein the composition is for preventing the occurrence of wrinkles in the skin, reducing the depth and / or number of wrinkles, and / or inhibiting an increase in the depth and / or number of wrinkles.

[61] The composition according to any of

[39] to

[60] , wherein the composition is for inhibiting or preventing skin darkening.

[62] The composition according to any of

[39] to

[61] , wherein the composition is for improving or preventing obesity.

[63] The composition according to any of

[39] to

[62] , wherein the composition is for slowing the progression of aging.

[64] The composition according to any one of

[39] to

[63] , which is a composition for extending lifespan.

[65] The composition according to any one of

[39] to

[64] , which is a composition for promoting plant growth.

[66] A method for treating a disease in a subject in need thereof, comprising administering to the subject a composition according to any one of [1] to

[28] or

[39] to

[65] .

[67] A method for ameliorating symptoms in a subject in need thereof, comprising administering to the subject a composition according to any one of [1] to

[28] or

[39] to

[65] .

[68] A compound of the following formula (I):

[69] Use of quinoid berberrubine represented by the following formula (I): in the preparation of a pharmaceutical composition.

[70] Use of quinoid berberrubine represented by the following formula (I): in the preparation of a food composition.

[71] Use of a quinoid type berberrubine represented by the following formula (I): in the preparation of a cosmetic composition.

[72] Use of a quinoid berberrubine represented by the following formula (I): in the preparation of an agricultural composition.

[73] A quinoid-type berberrubine represented by the following formula (I):

[74] A quinoid-type berberrubine represented by the following formula (I):

[75] A quinoid berberrubine represented by the following formula (I):

[76] Use of the berberrubine acetic acid adduct according to any one of

[33] to

[37] or the aqueous solution according to

[38] in the preparation of a pharmaceutical composition.

[77] Use of the berberrubine acetic acid adduct according to any one of

[33] to

[37] or the aqueous solution according to

[38] in the preparation of a cosmetic composition.

[78] Use of the berberrubine acetic acid adduct according to any one of

[33] to

[37] or the aqueous solution according to

[38] in the preparation of an agricultural composition.

[79] The berberrubine acetic acid adduct according to any one of

[33] to

[37] or the aqueous solution according to

[38] for use in the treatment of a disease in a subject in need thereof.

[80] The berberrubine acetic acid adduct according to any one of

[33] to

[37] or the aqueous solution according to

[38] for use in ameliorating symptoms in a subject in need thereof.

[81] The berberrubine acetic acid adduct according to any one of

[33] to

[37] or the aqueous solution according to

[38] for use for cosmetic purposes in a subject in need thereof.

[82] The berberrubine acetic acid adduct according to any one of

[33] to

[37] or the aqueous solution according to

[38] , for agricultural use in a subject in need thereof.

[0086] The present invention will be described below with reference to examples, but these examples are not intended to limit the scope of the present invention. Commercially available reagents, instruments, etc. referred to in the examples were used in accordance with the manufacturer's instructions or standard procedures unless otherwise specified.

[0087] Example 1 Preparation of Berberrubin-Related Compounds Commercially available berberine chloride (manufactured by Nacalai Tesque, Inc.) was heated at 190°C under reduced pressure for 3 hours to prepare quinoid berberrubine. The quinoid berberrubine thus prepared had the appropriate structure, as shown in Figures 1A and 1B, by solid analysis using the CPMAS and DDMAS methods. １３ The result was confirmed by C NMR. 400 mL of acetic acid was added to quinoid berberrubine prepared from 50.0 g of berberine chloride. Subsequently, after stirring at room temperature for 1 hour, 500 mL of diethyl ether was added. After stirring for another 3 hours, the residue was collected by suction filtration. The residue was dried at room temperature under reduced pressure for 2 hours to obtain berberrubine acetic acid adduct as orange-yellow crystals. The berberrubine acetic acid adduct was separated into solids by the CPMAS method and the DDMAS method, as shown in Figures 2A and 2B, respectively. １３ By C NMR, (i) the compound of the following formula (4): (ii) a compound represented by the following formula (5): [where n is 1 or more], or (iii) a mixture of compound (i) and compound (ii). That is, the berberrubine acetic acid adduct prepared in Example 1 is an acetic acid adduct of enol berberrubine and / or zwitterionic berberrubine. Here, n is the number of the solid １３ In C NMR, the equivalent of berberrubine was calculated from the integral of the peak of the 8-carbon atom of berberrubine at 26 ppm or 27-28 ppm, and the equivalent of acetic acid and / or acetate ion was calculated from the integral of the peak of the CH3 carbon atom of acetic acid at 20-25 ppm, and then the equivalent of acetic acid and / or acetate ion was calculated by dividing the equivalent of acetic acid and / or acetate ion by the equivalent of berberrubine. Furthermore, the berberrubine acetate adduct was further dried under reduced pressure at 80°C for 5 hours to obtain berberrubine acetate (berberrbine acetate adduct, where n = 1 in the above formulas (4) and (5)) as reddish-orange crystals. The structure of berberine acetate was obtained by CPMAS and DDMAS analysis of the solids shown in Figures 3A and 3B. １３The experimental values ​​of n calculated for the above-mentioned berberrubine acetic acid adduct and berberrubine acetate salt are shown in Table 1.

[0088]

[0089] Example 2: Solubility of Berberrubine-Related Compounds The solubility of quinoid-type berberrubine, berberrubine acetic acid adduct, and berberine acetate salt in water, methanol, dimethyl sulfoxide (DMSO), ethanol, and chloroform was measured. Measurements were performed according to the procedure described in the General Rules of the Japanese Pharmacopoeia, with some modifications. A solvent (water, methanol, dimethyl sulfoxide (DMSO), ethanol, or chloroform) was added to 1 g of a berberrubine-related compound (quinoid-type berberrubine, berberrubine acetic acid adduct, or berberine acetate salt), and the mixture was stirred for 30 seconds every 5 minutes at room temperature. After 30 minutes, the solution was checked for dissolution of each berberrubine-related compound. The minimum volume (mL) of solvent required to completely dissolve the berberrubine-related compound was determined by appropriately adjusting the amount of solvent. The solubility (g / mL) of the berberrubine-related compound in each solvent was calculated from the berberrubine-related compound (1 g) and the minimum volume (mL) of solvent used. The results are shown in Table 2.

[0090]

[0091] As shown in Table 2, the berberrubine acetic acid adduct was extremely soluble in water. The solubility of the berberrubine acetic acid adduct in water was approximately 36 times that of berberine acetate and approximately 2,300 times that of quinoid berberrubine.

[0092] High water solubility of a drug is important when administering or ingesting a drug to a living body, or when incorporating the drug into a composition containing the drug, such as a pharmaceutical composition, food composition, cosmetic composition, agricultural composition, etc. Therefore, the highly water-soluble berberrubine acetic acid adduct of the present disclosure is useful in these applications.

[0093] Example 3: Effects on mitochondrial basal respiration, ATP synthesis, and maximum respiration. A 3.5 mg / mL quinoid berberrubine DMSO solution was added to C2C12 cells in Dulbecco's modified Eagle's (DMEM) medium containing 10% fetal bovine serum (FBS), penicillin, and streptomycin to a final concentration of 30 μM (FIG. 4A). A 3.5 mg / mL aqueous solution of berberrubine acetic acid adduct was also added to the C2C12 cells to a final concentration of 30 μM (FIG. 4B). As a control, the same amount of water was added to the C2C12 cells. The time point 24 hours after the addition of the berberrubine acetic acid adduct was designated time 0 (minutes). An ATP synthase inhibitor (oligomycin, final concentration 10 μM) was added at 20 minutes, an uncoupler (carbonyl cyanide-m-chlorophenylhydrazone (CCCP), final concentration 10 μM) at 40 minutes, and a mitochondrial complex inhibitor (rotenone and antimycin, final concentration 10 μM) at 70 minutes were further added. During this time, the oxygen consumption rate (OCR) of C2C12 cells was measured over time using an extracellular flux analyzer.

[0094] The OCR during the period from the addition of the ATP synthase inhibitor to the addition of the uncoupler, the OCR during the period from the addition of the uncoupler to the addition of the mitochondrial complex inhibitor, and the OCR during the period after the addition of the mitochondrial complex inhibitor correlate with the amounts of basal mitochondrial respiration, ATP synthesis, and maximum respiration, respectively. Therefore, the OCR at 30, 60, and 90 minutes was used as an index to evaluate the amounts of basal mitochondrial respiration, ATP synthesis, and maximum respiration.

[0095] As shown in Figure 4C, addition of quinoid berberrubine significantly increased mitochondrial basal respiration, ATP synthesis, and maximal respiration by approximately 4-fold, compared to the control. Furthermore, as shown in Figure 4D, addition of the berberrubine acetate adduct significantly increased mitochondrial basal respiration, ATP synthesis, and maximal respiration by approximately 1.6-fold, 1.6-fold, and 1.5-fold, respectively.

[0096] These results indicate that the addition of both quinoid berberrubine and the berberrubine acetic acid adduct improves mitochondrial function, and therefore quinoid berberrubine and the berberrubine acetic acid adduct can have beneficial effects on diseases and symptoms caused by mitochondrial disorders.

[0097] Furthermore, the above results indicate that the addition of quinoid berberrubine improves mitochondrial ATP synthesis by approximately 2.5 times, maximal respiration by approximately 2.6 times, and basal respiration by approximately 2.5 times compared to the addition of berberrubine acetate adduct. Therefore, quinoid berberrubine can have more beneficial effects on diseases and symptoms caused by mitochondrial disorders than berberrubine acetate adduct.

[0098] Furthermore, the berberrubine acetic acid adduct used here is an acetic acid adduct of enol berberrubine and / or zwitterionic berberrubine, as described above. Furthermore, since the acetic acid (or acetate ion) contained in the berberrubine acetic acid adduct is liberated in the sample (water), the berberrubine acetic acid adduct acts as enol berberrubine and / or zwitterionic berberrubine. Therefore, compared with enol berberrubine and / or zwitterionic berberrubine, quinoid berberrubine can provide more advantageous effects against diseases and symptoms caused by mitochondrial disorders.

[0099] Example 4 Effect on the formation of mitochondrial respiratory chain supercomplexes C2C12 cells transfected with green fluorescent protein (GFP)-bound complex I and red fluorescent protein (RFP)-bound complex IV were cultured in Dulbecco's modified Eagle's (DMEM) medium containing 10% fetal bovine serum (FBS), penicillin, and streptomycin. A 3.5 mg / mL DMSO solution of quinoid berberrubine or a 3.5 mg / mL aqueous solution of berberrubine acetic acid adduct was added to the cells at a final concentration of 3, 10, 30, or 60 μM. After 24 hours, the formation of mitochondrial respiratory chain supercomplexes was detected by detecting FRET between GFP and RFP, which occurs when complex I and complex IV associate.

[0100] As shown in Figure 5, the addition of quinoid berberrubine or berberrubine acetic acid adduct promoted the formation of mitochondrial respiratory chain supercomplexes in a concentration-dependent manner.

[0101] The formation of mitochondrial respiratory chain supercomplexes is known to improve mitochondrial function, for example, by increasing the amount of energy produced by mitochondria and suppressing the production of reactive oxygen species (ROS) in cells. In other words, the above results indicate that quinoid-type berberrubine and berberrubine-acetic acid adducts improve mitochondrial function. Therefore, quinoid-type berberrubine and berberrubine-acetic acid adducts can provide beneficial effects against diseases and symptoms caused by mitochondrial dysfunction. Furthermore, quinoid-type berberrubine and berberrubine-acetic acid adducts can maintain cells in a healthy state by suppressing the production of reactive oxygen species (ROS), and can, for example, delay the progression of aging.

[0102] Example 5 Effect on Mouse Pituitary Cells Pituitary glands isolated from C57BL / 6NJcl mouse fetuses were minced with scissors in isolation buffer (0.123 M sodium chloride, 5 mM glucose, 5 mM potassium chloride, 1.3 mM calcium chloride, 100 mM HEPES-NaOH (pH 7.4), 4% BSA) containing 1 mg / mL collagenase, and the minced pituitary glands were heated in a water bath at 37°C for 30 minutes. After removing aggregates using a cell strainer, the cells were collected by centrifugation at 1,000 × g for 5 minutes and suspended in DMEM medium containing 20% ​​FBS to prepare primary cultures of pituitary cells. A 3.5 mg / mL aqueous solution of berberrubine acetic acid adduct was added to the primary cultured cells to a final concentration of 0, 30, 60, or 90 μM. After 48 hours of incubation after addition, mitochondrial proteins OPA1, HSP60, and Mfn2, as well as cellular proteins Tub (α-tubulin) or Actin (actin) were detected by Western blotting.

[0103] As shown in Figure 6A, the expression ratios of the mitochondrial proteins OPA1, HSP60, and Mfn2 relative to the cellular proteins Tub and actin increased in a concentration-dependent manner with berberrubine acetic acid adduct. The ratios of OPA1 to Tub expression (OPA1 / Tub), HSP60 to Tub expression (HSP60 / Tub), and Mfn2 to Actin expression (Mfn2 / Actin) reflect the mitochondrial content in cells. These results indicate that berberrubine acetic acid adduct concentration-dependently increases the mitochondrial content in cells. Meanwhile, the cell proliferation rate in primary cultured cells treated with berberrubine acetic acid adduct was comparable to that in primary cultured cells without berberrubine acetic acid adduct.

[0104] Furthermore, as shown in Figure 6B, when primary cultured cells to which the berberrubine acetic acid adduct had been added were stained with Hoechst for nuclei and Tom20 for mitochondria, and observed under a fluorescence microscope (magnification: 60x), no changes were observed compared to primary cultured cells to which the berberrubine acetic acid adduct had not been added, and no characteristics observed in cancer cells or senescent cells that are not found in healthy cells, such as enlarged nuclei, were observed.

[0105] These results indicate that berberrubine acetic acid adducts increase the mitochondrial content in cells without affecting healthy cell growth or morphology.

[0106] Example 6 Effect on Parkinson's Disease Mouse embryonic fibroblast (MEF) cells and PINK1-knockout MEF cells were cultured for 24 hours in low-glucose medium (Nacalai Tesque, Inc.) containing 10% fetal bovine serum (FBS), penicillin, and streptomycin. A 3.5 mg / mL DMSO solution of quinoid-type berberrubine or a 3.5 mg / mL aqueous solution of berberrubine acetate adduct was added to a final concentration of 30 μM. Forty-eight hours after addition, Tom20, a mitochondrial outer membrane protein, was detected by immunofluorescence staining. An anti-Tom20 antibody (Santa Cruz, sc-11415) was used to detect Tom20. PINK1 is a causative gene for Parkinson's disease, and PINK1 deficiency causes mitochondrial dysfunction associated with Parkinson's disease. Therefore, PINK1-knockout MEF cells can be used as a model for patients with Parkinson's disease. As shown in Figure 7, when quinoid berberrubine or berberrubine acetic acid adduct was administered to MEF cells, the intracellular mitochondrial content increased compared to controls (DMSO or water). Similarly, when quinoid berberrubine or berberrubine acetic acid adduct was added to PINK1-knockout MEF cells, the intracellular mitochondrial content also increased.

[0107] These results indicate that quinoid berberrubine and berberrubine acetic acid adducts can suppress mitochondrial dysfunction associated with Parkinson's disease by increasing the intracellular mitochondrial content, and therefore quinoid berberrubine and berberrubine acetic acid adducts can be effectively used in the treatment or prevention of Parkinson's disease.

[0108] Example 7 Inhibitory Effect on Cancer Cell Proliferation A 3.5 mg / mL quinoid berberrubine solution in DMSO or a 3.5 mg / mL aqueous solution of berberrubine acetate adduct was added to HeLa cells in Dulbecco's modified Eagle's (DMEM) medium containing 10% fetal bovine serum (FBS), penicillin, and streptomycin to a final concentration of 30 μM. 24 hours, 48 ​​hours, and 72 hours after addition, the cells were harvested, stained with trypan blue, and counted. HeLa cells are a cell line derived from human uterine cancer cells and are widely used as a cancer model cell line.

[0109] As shown in Figure 8A, the ratio of HeLa cell numbers after administration of quinoid berberrubine to control HeLa cell numbers decreased over time. In other words, the addition of quinoid berberrubine reduced the proliferation rate of HeLa cells compared to the control (DMSO). Also, as shown in Figure 8B, the ratio of HeLa cell numbers after administration of the berberrubine acetic acid adduct to control HeLa cell numbers decreased over time. In other words, the addition of an aqueous solution of the berberrubine acetic acid adduct reduced the proliferation rate of HeLa cells compared to the control (water).

[0110] These results indicate that quinoid berberrubine and berberrubine acetic acid adduct have the effect of suppressing the proliferation of cancer cells and are effective in treating or preventing cancer.

[0111] [Example 8] Effect on myelodysplastic syndrome (MDS) c-Kit cells collected from 7-week-old mice were infected with a retroviral vector incorporating a CBL mutant RNUX1 mutant co-introduced clone. These cells were transplanted via the tail vein into 7-week-old mice that had been sublethally irradiated, to generate hematopoietic stem cell transplant (BMT) mice with myelodysplastic syndrome (MDS).

[0112] As shown in Figure 9, the ratio of MDS clones (clones co-introduced with CBL mutation and RNUX1 mutation) to blood cell counts in BMT mice administered the berberrubine acetic acid adduct was significantly reduced compared to the control group (P<0.01). These results indicate that berberrubine acetic acid adduct is effective in treating or preventing cancer, including MDS.

[0113] Example 9 Effect on Expression and Phosphorylation of Neurological Disease-Related Proteins ΔN-P301L tau-CRY2olig was transfected into Neuro2A cells. One day after transfection, the cell cycle was arrested by the addition of nocotazole, and intracellular tau protein aggregation was induced by 24 hours of blue light irradiation. A 3.5 mg / mL quinoid-type berberrubine solution in DMSO or a 3.5 mg / mL aqueous solution of berberrubine acetic acid adduct was added to the tau protein-aggregated Neuro2A cells to a final concentration of 30 μM. After 8 hours of incubation, intracellular tau protein was detected by Western blotting using an antibody against S396-phosphorylated tau (pS396), an antibody against S422-phosphorylated tau (pS422), an antibody against full-length tau (JM), and an antibody against the C-terminal region of tau (tauC). The tau protein was detected at around 100 kDa.

[0114] As shown in Figure 10A, the addition of quinoid berberrubine reduced the expression level of S396-phosphorylated tau. More specifically, as shown in Figure 10C, the addition of quinoid berberrubine reduced the ratio of the expression level of S396-phosphorylated tau to the expression level of full-length tau (left graph in Figure 10C (vertical axis "pS396-tau / JM")) by approximately 35% compared to the control (administration of DMSO only) (p<0.01). On the other hand, no significant change was observed in the ratio of the expression level of S422-phosphorylated tau to the expression level of full-length tau (center graph in Figure 10C (vertical axis "pS422-tau / JM")). Furthermore, no significant change was observed in the expression level of full-length tau (right graph in Figure 10C). Phosphorylation of S396 of tau protein is known to be mediated by GSK3β. In other words, these results indicate that the addition of quinoid berberrubine inhibited the phosphorylation of tau protein by GSK3β.

[0115] Furthermore, as shown in Figure 10B, the addition of berberrubine acetic acid adduct decreased the expression level of S422-phosphorylated tau. More specifically, as shown in Figure 10D, the addition of berberrubine acetic acid adduct decreased the ratio of the expression level of S422-phosphorylated tau to the expression level of full-length tau (middle graph in Figure 10D (vertical axis "pS422-tau / JM")) by about 35% compared to the control (administration of water only) (p<0.01). On the other hand, no significant change was observed in the ratio of the expression level of S396-phosphorylated tau to the expression level of full-length tau (left graph in Figure 10D (vertical axis "pS396-tau / JM")). Furthermore, no significant change was observed in the expression level of full-length tau (right graph in Figure 10C).

[0116] It is known that phosphorylation of the S422 position of tau protein is mediated by JNK. Thus, these results indicate that the addition of berberrubine acetic acid adduct inhibited the phosphorylation of tau protein by the berberrubine acetic acid adduct.

[0117] Both phosphorylation of tau protein by GSK3β and phosphorylation of tau protein by JNK are known to cause neurological diseases, such as dementia (particularly Alzheimer's dementia). Therefore, both quinoid berberrubine and berberrubine acetate adducts can provide advantageous effects in the treatment or prevention of neurological diseases. Furthermore, the berberrubine acetate adduct used here is, as described above, an acetate adduct of enol berberrubine and / or zwitterionic berberrubine. Furthermore, since the acetic acid (or acetate ion) contained in the berberrubine acetate adduct is liberated in the sample (water), the berberrubine acetate adduct acts as enol berberrubine and / or zwitterionic berberrubine. Therefore, quinoid berberrubine can provide more advantageous effects on diseases and symptoms caused by mitochondrial disorders than enol berberrubine and / or zwitterionic berberrubine.

[0118] On the other hand, quinoid berberrubine and berberrubine acetic acid adducts (i.e., enol berberrubine and / or zwitterionic berberrubine) are thought to have different mechanisms of action because they inhibit the phosphorylation of tau protein. More specifically, quinoid berberrubine is thought to inhibit the activity of GSK3β, and berberrubine acetic acid adducts (i.e., enol berberrubine and / or zwitterionic berberrubine) are thought to inhibit the activity of JNK.

[0119] Example 10: Effect on sperm. Male C57BL / 6NJcl mice aged 9 to 10 weeks were orally administered 1.25% agar containing 0.35 mg / mL quinoid berberrubine or 1.25% agar without quinoid berberrubine for two weeks. Oral administration was performed by placing a container containing the agar in the mouse's cage, allowing the mouse to constantly ingest the agar from the container (hereinafter, oral administration of agar to mice was performed in the same manner). Mice at 20 weeks of age were put to sleep with isoflurane (Kanto Scientific) and then euthanized by cervical dislocation, and the testes and epididymis were removed. The removed epididymis was minced with scissors in 1 mL of phosphate-buffered saline (PBS) and allowed to stand at room temperature for 1 hour to prepare a sperm sample. 20 μL of the sample was transferred to a glass-bottom dish, and mitochondria and nuclei were stained with MitoTracker (Thermo Fisher Scientific) and Hoechst (Nacalai Tesque, Inc.), respectively.

[0120] Observation under a fluorescence microscope revealed that sperm in mice administered with quinoid berberrubine exhibited more active sperm motility than in controls. The observation results were recorded as a video. FIG. 11A is a snapshot of the recorded video. The stained sperm shown in FIG. 11A were more active in mice administered with quinoid berberrubine than in controls. Observation under a fluorescence microscope also revealed that the number of sperm whose mitochondria were stained with MitoTracker in mice administered with quinoid berberrubine increased approximately two-fold compared to controls (FIG. 11B). MitoTracker is a detection reagent that detects mitochondria depending on the mitochondrial membrane potential. It is also known that an increase in mitochondrial membrane potential is due to the activation of mitochondrial function. Therefore, these results indicate that quinoid berberrubine activated mitochondrial function.

[0121] Similarly, in mice orally administered with a 2 mg / mL aqueous solution of berberrubine acetic acid adduct, sperm motility was observed to be higher than in the control group. The observation results were recorded as a video. Figure 11C shows a snapshot of the recorded video.

[0122] These results indicate that quinoid-type berberrubine and berberrubine acetic acid adduct improve sperm motility and sperm mitochondrial function, and therefore, quinoid-type berberrubine and berberrubine acetic acid adduct can have beneficial effects in the treatment or prevention of infertility.

[0123] Example 11: Effect on skin Eight-week-old C57BL / 6NJcl mice were orally administered 1.25% agar containing 0.35 mg / mL quinoid berberrubine or 1.25% agar without quinoid berberrubine (control) for two months. During the same period, the mouse skin was irradiated with UV light three times a week for eight weeks. The irradiation dose was 1 mW / cm in the first week. ２ for 70 seconds (70 mJ / cm ２ ), and the second week was 1 mW / cm ２ for 80 seconds (80 mJ / cm ２ ), and the third week was 1 mW / cm ２ for 90 seconds (90 mJ / cm ２ ), and 1 mW / cm from the fourth week onwards ２ for 100 seconds (100 mJ / cm ２ ) At the end of the same period, the skin of the mice was visually observed. As shown in Figure 12, the mice administered with quinoid berberrubine had fewer wrinkles and shallower wrinkles than the mice not administered with quinoid berberrubine. Furthermore, the mice administered with quinoid berberrubine showed less darkening of the skin than the mice not administered with quinoid berberrubine.

[0124] These results indicate that quinoid berberrubine inhibits the formation of wrinkles in the skin, reduces the number and depth of wrinkles, and inhibits skin darkening.

[0125] Example 12 Effect on visceral fat reduction 50-week-old C57BL / 6NJcl mice were orally administered 1.25% agar containing 0.35 mg / mL quinoid berberrubine or 1.25% agar without quinoid berberrubine (control) for two months. At the end of the period, accumulation of visceral fat was observed in the mice. As shown in Figure 13, visceral fat accumulation was suppressed in mice administered with quinoid berberrubine compared to mice not administered with quinoid berberrubine.

[0126] These results indicate that quinoid berberrubine can suppress the accumulation of body fat, including visceral fat, and have beneficial effects in improving or preventing obesity.

[0127] Example 13 Effect on Brown Adipocytes Brown adipocytes isolated from C57BL / 6NJcl mouse fetuses were minced with scissors in isolation buffer (0.123 M sodium chloride, 5 mM glucose, 5 mM potassium chloride, 1.3 mM calcium chloride, 100 mM HEPES-NaOH (pH 7.4), 4% BSA) containing 1 mg / mL collagenase and heated in a water bath at 37°C for 30 minutes. After removing aggregates using a cell strainer, cells were collected by centrifugation at 1000 × g for 5 minutes and suspended in DMEM medium containing 20% ​​FBS to prepare primary cultures of brown adipocytes. A 3.5 mg / mL DMSO solution of quinoid berberrubine or a 3.5 mg / mL aqueous solution of berberrubine acetic acid adduct was added to the primary cultured cells to a final concentration of 30 μM. As a control, primary cultured cells were cultured for 48 hours with the same amount of DMSO as the quinoid berberrubine DMSO solution. After the addition, the primary cultured cells were stained with Hoechst (Nacalai Tesque) for nuclei and with tetramethylrhodamine methyl ester (TMRM, Thermo Fisher Scientific) for mitochondria, and then observed under a fluorescence microscope (magnification: 60x).

[0128] As shown in Figure 14, primary cultured cells treated with quinoid-type berberrubine and berberrubine acetate adduct showed strong mitochondrial staining by TMRM. TMRM is a cell-permeable fluorescent dye that accumulates in active mitochondria with normal membrane potential, and the signal becomes brighter when cells are healthy and have functioning mitochondria. Therefore, these results indicate that quinoid-type berberrubine and berberrubine acetate adduct increased active mitochondria with normal membrane potential in brown adipocytes. Furthermore, as shown in Figure 14, primary cultured cells treated with quinoid-type berberrubine and berberrubine acetate adduct, like the control, did not exhibit characteristics absent from healthy cells, such as nuclear enlargement, which are seen in cancer cells and senescent cells.

[0129] These results indicate that quinoid berberrubine and berberrubine acetic acid adducts have beneficial effects on diseases and symptoms caused by mitochondrial disorders by increasing active mitochondria. Furthermore, activation of mitochondrial function in brown adipocytes is known to reduce white adipocytes, such as subcutaneous fat. Therefore, these results indicate that berberrubine and berberrubine acetic acid adducts have beneficial effects on improving or preventing obesity.

[0130] Furthermore, as shown in Figure 14, the addition of quinoid berberrubine resulted in a greater increase in the amount of active mitochondria with normal membrane potential than the addition of the berberrubine acetic acid adduct. Therefore, quinoid berberrubine can have more advantageous effects on diseases and symptoms caused by mitochondrial disorders, or in the improvement or prevention of obesity, than the berberrubine acetic acid adduct.

[0131] Furthermore, the berberrubine acetic acid adduct used here is an acetic acid adduct of enol berberrubine and / or zwitterionic berberrubine, as described above. Furthermore, acetic acid (or acetate ions) is liberated from enol berberrubine and / or zwitterionic berberrubine in the sample (water). Therefore, compared with enol berberrubine and / or zwitterionic berberrubine, quinoid berberrubine can provide more advantageous effects on diseases and symptoms caused by mitochondrial disorders, or in the improvement or prevention of obesity.

[0132] Example 14 Effect on the Growth of Pea Seedlings Pea seedlings were sown in 10 cm Petri dishes containing a 0.035 mg / mL aqueous solution of the berberrubine-acetic acid adduct or water, and the growth of the seedlings was observed daily. As shown in Figure 15, the seedlings grown in the aqueous solution of the berberrubine-acetic acid adduct germinated more frequently and subsequently grew more rapidly than the seedlings grown in water.

[0133] These results indicate that the berberrubine acetic acid adduct has the effect of promoting plant growth and is useful as an agricultural composition.

[0134] Example 15 Effect on the Heart (1) From the age of 24 months onwards, C57BL / 6N24 mice were administered the berberrubine acetic acid adduct by allowing them to freely ingest a 0.5 mg / mL aqueous solution of the berberrubine acetic acid adduct or water (control) as drinking water.

[0135] The results of cardiac ultrasound examination by M-mode echocardiography on mice 12 weeks after the start of administration are shown in Figure 16. As shown in Figure 16, compared to controls, individuals administered the berberrubine acetic acid adduct had decreased left ventricular end-diastolic dimension (LVDd), left ventricular end-systolic dimension (LVDs), left ventricular diastolic function, and left ventricular filling pressure (E / e'), and increased left ventricular ejection fraction (LVEF) and % left ventricular fractional shortening (%FS). These results indicate that the berberrubine acetic acid adduct effectively improves cardiac function.

[0136] Furthermore, the autopsy findings for mice 12 weeks after the start of administration are shown in Figure 17. As shown in Figure 17, the ratio of heart weight to body weight was reduced in individuals administered the berberrubine acetic acid adduct compared to the control. On the other hand, no significant change was observed in the ratio of lung weight to body weight. These results indicate that the berberrubine acetic acid adduct is effective in improving and / or preventing cardiac hypertrophy.

[0137] Furthermore, the results of quantitative RT-PCR quantification of heart failure markers and myocardial fibrosis markers in mice 12 weeks after the start of administration are shown in Figure 18. As shown in Figure 18, the expression levels of heart failure markers (Nppa, Nppb, and Myh7) and myocardial fibrosis markers (Collagen1 and Ctgf) were reduced in individuals administered the berberrubine acetic acid adduct compared to controls. These results indicate that the berberrubine acetic acid adduct effectively reduces the risk of heart failure and myocardial fibrosis.

[0138] Furthermore, the expression of mitochondrial mRNA in mice 12 weeks after the start of administration was quantified by quantitative RT-PCR and the results are shown in Figure 19. As shown in Figure 19, the expression levels of mitochondrial mRNA (Pgc1a, Mitol, Tfam, Nrf1, Nrf2, Mfn1, Mfn2, Pink1, Park2, and Drp1) were increased in individuals administered the berberrubine acetic acid adduct compared to controls. These results indicate that the berberrubine acetic acid adduct activates mitochondria in the heart.

[0139] The above results indicate that berberrubine acetic acid adduct is effective in maintaining or improving healthy cardiac function, in improving or preventing cardiac hypertrophy, and in treating and / or preventing heart disease.

[0140] [Example 16] Effect on the Heart (2) C57BL / 6N24 mice were administered the following drinking water and food from the age of 24 months onwards. (1) Normal water as drinking water and normal food (CE-2, CLEA Japan, Inc., the same below) (control). (2) 0.5 mg / mL aqueous solution of berberrubine acetic acid adduct as drinking water and normal food as food (administration of berberrubine acetic acid adduct). (3) Normal water as drinking water and normal food containing 0.5 mg / mL quinoid berberrubine as food (administration of quinoid berberrubine).

[0141] The mice were examined by cardiac ultrasound using M-mode echocardiography 12 weeks after the start of administration, and the results are shown in FIG.

[0142] As shown in Figure 20, in both individuals administered berberrubine acetic acid adduct and quinoid berberrubine, compared to controls, left ventricular end-diastolic dimension (LVDd), left ventricular end-systolic dimension (LVDs), and an index representing left ventricular filling pressure (E / e') were reduced, and left ventricular ejection fraction (LVEF) and % left ventricular fractional shortening (%FS) were increased. These results indicate that berberrubine acetic acid adduct and quinoid berberrubine effectively improve cardiac function.

[0143] Furthermore, the results of quantitative RT-PCR quantification of heart failure markers and myocardial fibrosis markers in mice 12 weeks after the start of administration are shown in Figure 21. As shown in Figure 21, the expression levels of heart failure markers (Nppa, Nppb, and Myh7) and myocardial fibrosis markers (Collagen1 and Ctgf) were reduced in individuals administered the berberrubine acetic acid adduct and quinoid berberrubine compared to controls. These results indicate that the berberrubine acetic acid adduct and quinoid berberrubine effectively reduce the risk of heart failure and myocardial fibrosis.

[0144] Furthermore, the expression of mitochondrial mRNA, Mitol, in mice 12 weeks after the start of administration was quantified by quantitative RT-PCR and the results are shown in Figure 22. As shown in Figure 22, the expression level of Mitol increased in both mice administered the berberrubine acetic acid adduct and quinoid berberrubine compared to the control. These results indicate that the berberrubine acetic acid adduct and quinoid berberrubine effectively activate mitochondria in the heart.

[0145] In addition, mitochondria were isolated from mouse cardiomyocytes 12 weeks after the start of administration by standard methods, and the oxygen consumption rate (OCR) was measured using a flux analyzer. The results are shown in Figures 23A and 23B. As shown in Figures 23A and 23B, the oxygen consumption rate (OCR) of the berberrubine acetic acid adduct and quinoid berberrubine was elevated compared to the control in all three states: State 2 (ADP-limited state; indicating basal mitochondrial respiration rate), State 3 (activated respiratory state; indicating mitochondrial energy production capacity), State 4o (oligomycin-induced state; indicating mitochondrial non-phosphorylated oxygen consumption (proton leak)), and State 3u (uncoupled state; indicating mitochondrial maximum oxygen consumption capacity). These results indicate that the berberrubine acetic acid adduct and quinoid berberrubine effectively activate mitochondria in the heart.

[0146] The above results indicate that berberrubine acetic acid adduct and quinoid berberrubine are effective in maintaining or improving healthy cardiac function, in improving or preventing cardiac hypertrophy, and in treating and / or preventing heart disease.

[0147] Example 17 Effect on Mitochondrial Encephalomyopathy, Lactic Acidosis, and Stroke-like Episodes Syndrome (MELAS) Human dermal fibroblast (NHDF) cells and mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes syndrome (MELAS)-derived cells F954 (3243A>G, 92%) and F969 (3243A>G, 34.5%) were cultured on day 1, with the analysis starting day designated as day 0. An aqueous solution of the berberrubine acetic acid adduct or water was added to a final concentration of 30 μM to the cell culture medium, and oxygen consumption rate (OCR) was measured on day 8 using a flux analyzer.

[0148] As shown in Figures 24A and 24B, when an aqueous solution of berberrubine acetic acid adduct was added, the basal respiration, maximum respiration, and spare respiratory capacity of NHDF cells, F954 cells, and F969 cells increased compared to when water was added, indicating enhanced mitochondrial function. The degree of increase was greater in MERAS-derived cells, F954 cells and F969 cells, compared to NHDF cells. These results indicate that berberrubine acetic acid adduct activates mitochondria and is particularly effective in the treatment and / or prevention of MERAS.

[0149] Example 18 Effect on Inflammatory Cancer The effect of the berberrubine acetic acid adduct on inflammatory cancer was analyzed using a mouse inflammatory carcinogenesis model. Specifically, the berberrubine acetic acid adduct was administered to 57BL / 6 mice from 5 weeks of age onward by allowing them to freely consume a 1 mg / mL aqueous solution of the berberrubine acetic acid adduct or water as drinking water. Seven days after the start of administration, C57BL / 6 mice were co-implanted with regressed QR-32 cells and a gelatin sponge subcutaneously into the right flank of the mouse body according to the method described in Okada et al., Br. J. Cancer, Vol. 66, pp. 635-639 (1992), to allow tumor formation. The size of the tumors formed in the mice after co-implantation was observed.

[0150] As shown in Figure 25, 36 weeks after co-transplantation, tumors remained in 5 of 7 mice (71%) that had been given water as drinking water. In contrast, 36 weeks after co-transplantation, tumors disappeared in all mice that had been given an aqueous solution of the berberrubine acetic acid adduct as drinking water (0 of 6 mice (0%)).

[0151] These results indicate that the berberrubine acetic acid adduct effectively inhibits inflammatory cancer. Similar analyses are possible for the effects of quinoid berberrubine on inflammatory cancer.

[0152] Example 19 Lifespan Extension Effect From 8 weeks of age onwards, mice were fed a high-fat diet (High Fat Diet 32, CLEA Japan, Inc.), and from 60 weeks of age onwards, they were allowed to freely drink a 0.5 mg / mL aqueous solution of berberrubine acetic acid adduct or water (control), and then their survival time was analyzed.

[0153] As shown in Figure 26, administration of the berberrubine acetic acid adduct extended survival time compared to the control. That is, all 13 control individuals tested at 92 weeks of age died (survival rate: 0%). On the other hand, of the individuals administered with the berberrubine acetic acid adduct, 13 of 14 individuals tested at 92 weeks of age (survival rate: 93%) survived. Furthermore, 50% survival was achieved at 75 weeks of age in the control individuals, whereas it was achieved at 105 weeks of age in the individuals administered with the berberrubine acetic acid adduct. That is, the berberrubine acetic acid adduct extended the 50% survival rate by 30 weeks.

[0154] These results indicate that the berberrubine acetic acid adduct effectively extends the lifespan.

[0155] The disclosures of all patents, patent applications and publications cited in this disclosure are hereby incorporated by reference in their entireties.

[0156] The composition of the present disclosure can be suitably used as, for example, a pharmaceutical industrial product, a food industrial product, a cosmetic industrial product, or an agricultural industrial product. Therefore, the present invention has industrial applicability.

Claims

1. A compound of formula (I): A composition comprising a quinoid type berberrubine represented by the formula:

2. The composition of claim 1, wherein the compound represented by formula (I) is an isolated quinoid type berberrubine.

3. The composition of claim 1 which is a pharmaceutical composition.

4. The composition of claim 1 which is a food composition.

5. The composition of claim 1 which is a cosmetic composition.

6. The composition of claim 1, which is an agricultural composition.

7. The composition according to claim 1, which is a composition for activating mitochondria.

8. The composition according to claim 1, which is a composition for treating or preventing a mitochondrial disease.

9. The composition according to claim 1, which is a composition for treating or preventing Parkinson's disease.

10. The composition according to claim 1, which is a composition for treating or preventing mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes syndrome (MELAS).

11. The composition according to claim 1, which is a composition for treating or preventing cancer.

12. The composition of claim 11, wherein the cancer is an epithelial cancer, carcinoma, cervical cancer, sarcoma, or myelodysplastic syndrome (MDS).

13. The composition according to claim 1, which is a composition for treating or preventing a muscle disease.

14. The composition according to claim 13, wherein the muscle disease is frailty, locomotive syndrome, or sarcopenia.

15. The composition according to claim 1, which is a composition for treating or preventing a neurological disease.

16. The composition according to claim 15, wherein the neurological disease is dementia.

17. The composition according to claim 1, which is a composition for treating or preventing heart disease.

18. The composition according to claim 17, wherein the heart disease is heart failure.

19. The composition according to claim 17, wherein the heart disease is cardiomyopathy.

20. The composition according to claim 1, which is a composition for improving or preventing cardiac hypertrophy.

21. The composition according to claim 1, which is a composition for improving infertility.

22. The composition according to claim 1, which is a composition for increasing sperm activity.

23. The composition according to claim 1, which is a composition for preventing the occurrence of wrinkles in the skin, for reducing the depth and / or number of wrinkles, and / or for inhibiting the increase in the depth and / or number of wrinkles.

24. The composition according to claim 1, which is a composition for inhibiting or preventing skin darkening.

25. The composition according to claim 1, which is a composition for improving or preventing obesity.

26. The composition of claim 1, which is a composition for slowing the aging process.

27. The composition of claim 1, which is a composition for extending life.

28. The composition of claim 1, which is a composition for promoting plant growth.

29. A compound represented by the following formula (I): A method for producing a berberrubine acetic acid adduct, comprising adding acetic acid to a quinoid type berberrubine represented by the following formula:

30. The process of claim 29, comprising adding acetic acid as acetic acid vapor.

31. (i) A compound represented by the following formula (I): (ii) adding acetic acid to a quinoid type berberrubine represented by the formula:

32. The process according to claim 31, wherein in (i), acetic acid is added as acetic acid vapor to produce a berberrubine acetic acid adduct.

33. Berberrubine acetate adduct.

34. The following equation (2): or an enol type berberrubine represented by the following formula (3): The berberrubine acetic acid adduct according to claim 33, wherein acetic acid and / or acetate ions are added to a zwitterionic berberrubine represented by the formula:

35. The berberrubine acetic acid adduct according to claim 33, in which one or more equivalents of acetic acid and / or acetate ions are added to one equivalent of berberrubine.

36. (i) The following formula (4): (ii) a compound represented by the following formula (5): The berberrubine acetic acid adduct according to claim 33, which is a compound represented by the formula (5): [wherein n is 1 or more]; or (iii) a mixture of compound (i) and compound (ii).

37. The berberrubine acetic acid adduct according to claim 33, having a solubility in water of 0.01 g / mL or more.

38. An aqueous solution of the berberrubine acetic acid adduct described in claim 33.

39. A composition comprising the berberrubine acetic acid adduct of claim 33 or the aqueous solution of claim 38.

40. The composition of claim 39 which is a pharmaceutical composition.

41. The composition of claim 39 which is a food composition.

42. The composition of claim 39 which is a cosmetic composition.

43. The composition of claim 39 which is an agricultural composition.

44. The composition of claim 39, which is a composition for activating mitochondria.

45. The composition of claim 39, which is a composition for treating or preventing a mitochondrial disease.

46. ​​The composition of claim 39, which is for the treatment or prevention of Parkinson's disease.

47. The composition according to claim 39, which is a composition for treating or preventing mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes syndrome (MELAS).

48. The composition of claim 39, which is a composition for treating or preventing cancer.

49. The composition of claim 48, wherein the cancer is an epithelial cancer, carcinoma, cervical cancer, sarcoma, or myelodysplastic syndrome (MDS).

50. The composition of claim 39, which is for treating or preventing a muscle disease.

51. The composition described in claim 50, wherein the muscle disease is frailty, locomotive syndrome, or sarcopenia.

52. The composition according to claim 39, which is for treating or preventing a neurological disease.

53. The composition of claim 52, wherein the neurological disease is dementia.

54. The composition of claim 39, which is for the treatment or prevention of heart disease.

55. The composition of claim 54, wherein the heart disease is heart failure.

56. The composition of claim 54, wherein the heart disease is cardiomyopathy.

57. The composition of claim 39, which is a composition for improving or preventing cardiac hypertrophy.

58. The composition of claim 39, which is a composition for improving infertility.

59. The composition described in claim 39, which is a composition for increasing sperm activity.

60. The composition of claim 39, which is a composition for preventing the occurrence of wrinkles in the skin, for reducing the depth and / or number of wrinkles, and / or for inhibiting the increase in the depth and / or number of wrinkles.

61. The composition of claim 39, which is a composition for inhibiting or preventing skin darkening.

62. The composition according to claim 39, which is a composition for improving or preventing obesity.

63. The composition of claim 39, which is a composition for slowing the aging process.

64. The composition of claim 39, which is a composition for extending life.

65. The composition of claim 39, which is a composition for promoting plant growth.