JP7680768B2 - Composition containing ingredients derived from Japanese pepper or connarus - Google Patents

Description

Patent Law Article 30, Paragraph 2 applied. Published at the Abstracts of the 58th Japanese Society of Nutrition and Food Science Kinki Branch Conference on October 21, 2019.

Application of Article 30, Paragraph 2 of the Patent Act. Presented at the 58th Kinki Branch Meeting of the Japanese Society of Nutrition and Food Science on November 16, 2019.

Application of Article 30, Paragraph 2 of the Patent Act February 3, 2020, Announced in the Master's Thesis Abstract of the Graduate School of Health and Nutrition, Kobe Women's University, 2019

Application of Article 30, Paragraph 2 of the Patent Act. Presented at the Master's Thesis Presentation Session of the Graduate School of Health and Nutrition, Kobe Women's University, on February 3, 2020.

Article 30, paragraph 2 of the Patent Act applies. Published on February 3, 2020, in the Master's thesis "Study on the preventive effect of grape sansho extract on muscle atrophy" at the Graduate School of Health and Nutrition, Kobe Women's University, 2019.

This application relates to a composition containing ingredients derived from Japanese pepper or connarus.

The aging of the population is progressing in developed countries around the world. In order for the elderly to stay healthy both physically and mentally, it is important to maintain muscle and bones. However, in the elderly, muscle and bone loss occurs not only due to aging, but also as a symptom of disuse syndrome caused by reduced physical activity or long-term inactivity. Therefore, a composition for strengthening muscle and bones, especially in the elderly, was needed.

The inventors discovered that Japanese pepper, a spice that has long been used as a food and herbal medicine in the Orient, and Connarus arvensis, a plant of the Bucconaceae family native to Brazil, are useful for preventing or treating disuse syndrome, and thus completed this invention.

The present application provides, for example, the following items:
(Item 1)
A composition containing an ingredient derived from the plant body of Zanthoxylum japonicum or Connarus japonicus for building muscle or bone.
(Item 2)
2. The composition of claim 1, wherein the plant matter is selected from the group consisting of fruits, inner bark, mature leaves, and seeds.
(Item 3)
3. The composition according to item 1 or 2, wherein the Japanese pepper is Budou Japanese pepper or Asakura Japanese pepper.
(Item 4)
The composition according to any one of items 1 to 3, wherein the ingredient is a crushed plant body of Zanthoxylum japonicum or Connarus japonicum.
(Item 5)
The composition according to any one of items 1 to 4, wherein the ingredient is derived from an extract obtained by extracting a plant body of Zanthoxylum japonicum or Zanthoxylum conulum with a solvent containing water.
(Item 6)
The composition according to any one of items 1 to 5, wherein the ingredient is a concentrate or a dried product of an extract obtained by extracting a plant body of Zanthoxylum japonicum or Zanthoxylum conulum with a solvent containing water.
(Item 7)
7. The composition according to any one of items 1 to 6, for preventing or treating a disease selected from the group consisting of diet-induced bone atrophy, disuse syndrome, aging-related bone atrophy, and postmenopausal osteoporosis.
(Item 8)
The composition according to any one of items 1 to 7, wherein the disease is disuse syndrome.
(Item 9)
9. The composition according to any one of items 1 to 8, which is intended to be taken orally.
(Item 10)
10. A food product comprising the composition according to any one of items 1 to 9.
(Item 11)
A method for producing the composition according to any one of items 1 to 9, comprising the steps of:
The production method includes a step of extracting crushed plant bodies of Zanthoxylum japonicum or Zanthoxylum conulum with a solvent containing water.

A cross-section of a Japanese pepper tree, showing the location of the inner bark. Freeze-dried samples. A method for creating a rat model of disuse syndrome. The rats were suspended from the floor with their tails clipped, and disuse syndrome was induced in the hind limbs by raising them without allowing their hind limbs to touch the floor. Effects of hot water extract of fruit or inner bark of Japanese pepper on body weight of disuse syndrome model rats. Values are expressed as mean ± SD for 6-7 rats. Tukey's test with a significance level of 5% was performed on other groups. *: p<0.05 (vs. water-hindlimb suspension (-) group). Water intake of disuse syndrome model rats given water or hot water extract of Japanese pepper. Values are expressed as mean ± SD for 6-7 rats. Tukey's test with a significance level of 5% was performed on other groups. Effects of hot water extract of fruits or inner bark of Japanese pepper on soleus muscle weight in disuse syndrome model rats. Values are expressed as mean ± SD for 6-7 rats. Tukey's test with a significance level of 5% was performed on other groups. *: p<0.05 (vs. water-hindlimb suspension (-) group). Cross-sectional image of HE-stained soleus muscle (x10). Effects of hot water extracts of Budo Sansho, Asakura Sansho, or Connarus on bone strength in rats with disuse syndrome. The a and b above the error bars indicate significant differences at a risk level of 5% compared to the Cont(-) group (water-hindlimb suspension (-) group) or the Cont(+) group (water-hindlimb suspension (+) group), respectively.

[Definition of terms]
It should be understood that the terms used in this specification are used in the sense commonly used in the art unless otherwise specified. Therefore, unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by those skilled in the art to which this application belongs. In case of conflict, the specification of this specification (including definitions) shall prevail.

In this specification, "about" refers to ±10% of the indicated value, unless otherwise specified.

In this specification, "sansho" refers to a deciduous shrub of the Rutaceae family that has a unique fragrance, mainly in its leaves and fruit, and is used as a spice. In addition, in this specification, "connarus" refers to plants of the genus Connarus in the family Matricaria, such as Connarus rube (scientific name: Connarus ruber, also known as Connarus ruber).

In this specification, "bone atrophy" refers to a state in which bone tissue is reduced. It occurs when the bone resorption mechanism is higher than normal, or when the bone addition mechanism is lower than normal. In this specification, "muscle atrophy" refers to a decrease in muscle volume. Muscle atrophy occurs due to atrophy or a decrease in the number of muscle fibers that make up the muscle, or due to degeneration or damage to the motor nerves that control the muscle.

In this specification, "disuse syndrome" refers to a group of various symptoms resulting from a state of reduced physical activity, and therefore "disuse bone atrophy" and "disuse muscle atrophy" refer to bone atrophy and muscle atrophy resulting from a state of reduced physical activity.

In this specification, "postmenopausal osteoporosis" refers to a condition of bone loss that occurs rapidly in women after menopause, and is associated with a decrease in secretion of the female hormone estrogen.

[Embodiment]
Preferred embodiments of the present invention will be described. It should be understood that the embodiments provided below are provided for a better understanding of the present invention, and are not intended to limit the scope of the present invention to the following description.

(composition)
In one aspect, the present application provides a composition for muscle or bone enhancement, comprising an ingredient derived from the plant body of Zanthoxylum japonicum or Connarus japonicum. In one embodiment, muscle or bone enhancement may be suppression of muscle or bone mass loss, or may be increase of muscle or bone mass. Muscle or bone enhancement may be suppression of deterioration of muscle or bone quality, or may be improvement of muscle or bone quality. The mechanism of muscle or bone enhancement may be reduction of decomposition and absorption of muscle or bone, or promotion of muscle or bone formation.

In one embodiment, the present application provides a composition containing an ingredient derived from a plant body. In this specification, the ingredient derived from a plant body refers not only to a specific compound contained in the plant body, but also to any form of substance obtained by processing or treating the raw plant body. In one embodiment, the ingredient derived from a plant body includes a substance obtained by subjecting the plant body to drying, crushing, extraction, concentration, or any combination of these treatments. The ingredient derived from a plant body may contain an active ingredient in the plant body to achieve the desired effect of the present application. In one embodiment, the plant body may include any part or combination of a plant, such as petals, stamens, pistils, sepals, seeds, buds, outer bark, cork tissue, inner bark, leaves, guard cells, palisade tissue, spongy tissue, meristematic tissue, reticulum, inner coat, and vascular bundles (xylem, phloem, etc.). In one embodiment, components derived from a plant include, but are not limited to, plant cell walls, cell membranes, cytosols, mitochondria, chloroplasts, plastids, nuclear membranes, endoplasmic reticulum, vacuoles, nuclei, cytoskeleton, Golgi apparatus, and vesicles.

In one embodiment, the plant may be a Japanese pepper plant, and Japanese pepper may be a plant of the genus Zanthoxylum in the family Rutaceae. The lineages of the Zanthoxylum genus Zanthoxylum include Zanthoxylum piperitum (L.) DC., Zanthoxylum piperitum (L.) DC. f. brevispinum (Makino) Makino, Zanthoxylum piperitum (L.) DC. f. hispidum Hayashi, Zanthoxylum piperitum (L.) DC. f. inerme (Makino) Makino, also written as "Asakura Zanthoxylum", Zanthoxylum piperitum (L.) DC. f. ovatifoliolatum (Nakai) Makino, Zanthoxylum piperitum (L.) DC. f. rotundatum ... Examples of zansho include, but are not limited to, Asakura zansho or Budo zansho, which is derived from Asakura zansho, and Takahara zansho which grows wild in the Takahara River basin in Gifu Prefecture. Preferred are Asakura zansho or Budo zansho. Furthermore, other species of plants in the genus Zanthoxylum include Zanthoxylum ailanthoides Siebold et Zucc., Zanthoxylum ailanthoides Siebold et Zucc. f. pubescens (Hatus.) H.Ohba, and Zanthoxylum ailanthoides Siebold et Zucc. var. ailanthoides f. espinosum Zanthoxylum amamiense Ohwi, Zanthoxylum armatum DC. var. armatum, Zanthoxylum armatum DC. var. subtrifoliatum (Franch.) Kitam. Zanthoxylum beecheyanum K.Koch var. alatum (Nakai) H.Hara, Zanthoxylum beecheyanum K.Koch var. beecheyanum, Zanthoxylum bungeanum Maxim., Zanthoxylum dimorphophyllum Hemsl., Zanthoxylum fauriei (Nakai) Ohwi, Zanthoxylum integrifoliolum (Merr.) Merr., Zanthoxylum nitidum (Roxb.) DC., Zanthoxylum pteropodum Hayata, Zanthoxylum scandens Blume, Zanthoxylum schinifolium Siebold et al. Zucc.), Zanthoxylum schinifolium Siebold et Zucc. var. inerme (Nakai) T.B.Lee, Zanthoxylum schinifolium Siebold et Zucc. var. okinawense (Nakai) Hatus. ex Shimabuku, Zanthoxylum simulans Hance, Zanthoxylum yakumontanum (Sugim.) Nagam., and Zanthoxylum wutaiense I.S.Chen may be used. The common edible parts of Japanese pepper are the fruit and leaves (known as "kinome"), but in the present application, either the edible or inedible parts may be used.

In one embodiment, the plant is a Connarus plant. Connarus, as defined above, refers to a plant of the genus Connarus in the family Mammalianaceae, and examples thereof include Connarus rube (scientific name: Connarus ruber, also called Connarus ruber). The Connarus plant that can be used in the composition of the present application is not particularly limited, and for example, above-ground parts, roots, or mixtures thereof can be used, but among these, it is preferable to use above-ground parts in order to increase production efficiency. As the above-ground parts, it is preferable to use leaves, branches, bark, stems, fruits, etc., and it is particularly preferable to use bark.

In one embodiment, the components derived from the plant body can be obtained by subjecting the plant body to operations such as drying, crushing, cutting, dissolving, decomposing, denaturing, powdering, or extraction. For example, after drying, Japanese pepper or conalus may be crushed using a coarse crusher, or may be crushed without drying. Drying may be performed in the sun, using a dryer, or by freeze-drying. Known drying methods include natural drying, heat drying, water-absorbent drying, airflow drying, reduced pressure drying, vacuum drying, freeze-drying, high-frequency drying, air-conditioning drying, freeze-drying, and spray drying. In addition, the plant body may be subjected to a pretreatment such as degreasing with a non-polar solvent such as hexane, and then crushed to obtain the components derived from the plant body. The size adjustment by crushing is known to those skilled in the art, and may be roughly crushed to about several mm, may be crushed to several hundred μm, or may be finely crushed to about 10 μm so that it does not feel rough when dissolved in water and drunk.

The crushed material obtained may be classified by sieving using particle size, or by classification using the difference in the moving distance against wind or centrifugal force. The particle size of the powder obtained by classification may be such that the entire amount passes through a No. 18 (850 μm) sieve and 5% or less of the total amount remains on a No. 30 (500 μm) sieve, or it may be finer, with 10% or less of the total amount passing through a No. 200 (75 μm) sieve. In addition, only specific plant bodies may be selected and crushed during the crushing process, or only specific plant bodies may be selected after crushing. In one embodiment, the composition of the present application or the components derived from the plant of the present application may contain water, and for example, the moisture content may be 15% by weight or less, 10% by weight or less, 5% by weight or less, etc. In one embodiment, the composition of the present application or the component derived from the plant body of the present application may be granules made by crushing powder into a granular form, and additives such as binders and disintegrants may be added when making the granules. Binders and disintegrants used in granulation are known to those skilled in the art, and include cellulose and its derivatives (such as crystalline cellulose, hydroxypropyl cellulose, and carmellose), starch and its derivatives (such as starch, hydroxypropyl starch, dextran, and dextrin), natural polymer compounds (such as gum arabic, agar, and sodium alginate), and synthetic polymers (such as povidone and polyvinyl alcohol).

In one embodiment, the components derived from the plant body can be obtained by extracting them from the plant body (e.g., crushed matter). As the extraction solvent, it is preferable to use a polar solvent, such as water, an aqueous solvent, a hydrophilic organic solvent, etc., and it is preferable to extract them alone or in combination of two or more at room temperature or at a temperature below the boiling point of the solvent. Water that can be used as the extraction solvent includes pure water, tap water, well water, mineral water, mineral water, hot spring water, spring water, fresh water, etc., as well as water that has been subjected to various treatments. Treatments that can be applied to water include, for example, purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, and buffering. Therefore, water that can be used as the extraction solvent in this application also includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered physiological saline, supercritical water, and subcritical water. In addition, an aqueous solvent refers to an aqueous solution in which substances other than hydrophilic organic solvents (e.g., water-soluble salts such as table salt) are dissolved in such water at a concentration that does not lose the properties of the extraction solvent. The extraction solvent is preferably water, more preferably hot water.

The obtained extract may be subjected to treatment such as dilution, concentration, drying, or purification according to a conventional method, to obtain a concentrate of the extract, a dried product, or a crude or purified product thereof. Purification can be performed, for example, by activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, etc., and ion exchange resin treatment is particularly preferable. The obtained extract may be used as it is as the composition of the present application, but may also be made into a concentrate or dried product. In one embodiment, the drying method is known to those skilled in the art, and includes natural drying, heat drying, water absorbent drying, air flow drying, reduced pressure drying, vacuum drying, freeze drying, high frequency drying, air conditioning drying, freeze drying, and spray drying. An example of the drying method is a method in which an excipient such as dextran or dextrin is added to the extract, and then the extract is dried by a vacuum drying method, a freeze drying method, or a spray drying method. The dried product obtained by such a drying method may be dissolved in water again. In one embodiment, the concentration method is known to those skilled in the art, and includes heat concentration, reduced pressure concentration, spray concentration, ventilation concentration, and freezing concentration. The concentrate obtained by such a concentration method may be diluted again with water.

In one embodiment, the composition of the present application may be used to prevent or treat a disease selected from the group consisting of diet-induced bone atrophy, disuse syndrome (such as disuse bone atrophy and disuse muscle atrophy), aging bone atrophy, and postmenopausal osteoporosis. In one embodiment, the composition of the present application may be used to prevent or treat disuse syndrome. In a specific embodiment, the composition of the present application may be used to prevent or treat disuse bone atrophy and/or disuse muscle atrophy. Disuse bone atrophy and disuse muscle atrophy are symptoms of disuse syndrome, and are seen in people whose activity has decreased due to mental or physical disorders, or in hospitalized patients who cannot move their entire body or part of it due to cast immobilization or bed rest instructions, and are particularly seen in elderly people who have become less active due to a decline in physical ability. In addition, bone and muscle atrophy may occur in people who work mainly at a desk and have little physical activity, and in people who spend long periods in low-gravity conditions (e.g., underwater and in space) where stimulation to muscles and bones is reduced. Disuse syndrome caused by reduced activity leads to a decline in physical abilities such as physical strength, which leads to further decline in activity. Elderly people are prone to this vicious cycle, and it is important to prevent disuse syndrome before it occurs.

In one embodiment, the composition of the present application can be used to prevent or treat diet-induced bone atrophy. Diet-induced bone atrophy is bone atrophy caused by nutritional deficiency, particularly calcium insufficiency. Calcium insufficiency can occur not only due to a lack of calcium in the diet, but also due to insufficient absorption of calcium in the intestine or excretion of absorbed calcium. Vitamin D is necessary for calcium absorption in the intestine, but in order to synthesize vitamin D in the body, it is necessary to receive ultraviolet rays contained in sunlight. Therefore, if ultraviolet rays are prevented from reaching the skin by sunscreen, etc., vitamin D may be insufficient and calcium absorption may be insufficient. In addition, smoking weakens the intestine's nutrient absorption function, and excessive intake of caffeine, alcohol, or salt increases calcium excretion, causing calcium deficiency.

In one embodiment, the subject in need of administration of the composition of the present application is determined, for example, by the following parameters. According to the Ministry of Health, Labor and Welfare, the following physical activity goals for health are recommended: (1) 40 minutes of physical activity per day for those aged 65 or older, (2) 60 minutes of physical activity per day and 60 minutes of exercise per week for those aged 18 to 64, and (3) 60 minutes or more of exercise per day for those under 18. In one embodiment, the subject is a subject who is only engaged in physical activity below the above goals. Without wishing to be bound by a particular theory, it is said that in the human body, if a specific muscle is not used completely, 1 to 3% of muscle strength will decrease in one day, 10 to 15% in one week, and about 50% in one month. In one embodiment, the subject is a subject who does not move a muscle (e.g., a specific muscle) for a specific period of time, such as one day, one week, or one month, or who is at risk of not moving the muscle.

Furthermore, the diagnostic criteria for primary osteoporosis set out by the Japanese Society for Bone and Mineral Research and the Japanese Osteoporosis Society state that (1) a patient has a fragility fracture (a fracture caused by a minor external force such as a fall from a standing position) in the vertebral body or proximal femur and has a bone mineral density that is less than 80% of the YAM (young adult average: the average bone mineral density in the hip bone in people aged 20-44 or the proximal femur in people aged 20-39), or (2) a patient has no fragility fracture and has a bone mineral density that is 70% or less of the YAM or -2.5 SD or less. Therefore, in one embodiment, the subject meets the above diagnostic criteria for primary osteoporosis. According to the guideline for determining vitamin D insufficiency/deficiency set by the Japan Society for Bone and Mineral Research and the Japan Osteoporosis Society, in measuring vitamin D concentration based on serum 25(OH)D value, (1) a serum 25(OH)D value of 30 ng/mL or more is deemed to be vitamin D sufficient, (2) a serum 25(OH)D value of 20 ng/mL or more and less than -30 ng/mL is deemed to be vitamin D insufficient, and (3) a serum 25(OH)D value of less than 20 ng/mL is deemed to be vitamin D deficient. As described above, vitamin D is an essential nutrient for calcium intake in the body, and it has been reported that a group of subjects in a vitamin D deficient state has a significantly higher number of osteoporosis subjects than a group of subjects in a vitamin D deficient or sufficient state. Therefore, one of the subjects who require administration of the composition of the present application is a subject who falls into the above vitamin D deficient or insufficient state, and particularly a subject who falls into a vitamin D deficient state.

In one embodiment, the composition of the present application may be used to prevent or treat aging bone atrophy. It is generally known that in humans, bone mass reaches its maximum in the 20s, and bone mass begins to decrease from the 40s. This is thought to be due to an imbalance between old bone resorption and new bone formation. In particular, it is known that postmenopausal women are more likely to develop postmenopausal osteoporosis, in which bone atrophy occurs due to a decrease in the secretion of estrogen, also known as the female hormone, and the inhibition of bone resorption by estrogen is released. In one embodiment, the composition of the present application is intended for ingestion by humans (e.g., women) aged 40 or older. In one embodiment, the composition of the present application is intended for ingestion by postmenopausal women. In one embodiment, the composition of the present application may also be used to prevent or treat postmenopausal osteoporosis.

It is known that Conarus extract prevents the glycation of proteins, including collagen. Without wishing to be bound by any particular theory, it is believed that Conarus extract inhibits the glycation of collagen in bones, thereby inhibiting bone resorption, since bones are made of collagen and calcium.

(Food)
In one embodiment, the composition of the present application may be used in food for human consumption. Therefore, in one embodiment, the composition of the present application may be prepared to remove, decompose, denature or neutralize plant parts and/or plant components that are repulsive to human consumption due to taste, smell, texture or hygiene. For example, the outer bark of the plant is hard and may be unsuitable as food, so it may be removed. For example, components such as lignin contained in the plant may be bitter and may be unsuitable as food, so it may be removed. Those skilled in the art may remove plant parts and/or plant components by any suitable method. The composition of the present application may be provided as a food by itself, or may be provided as a food in combination with other food ingredients. In one aspect, the present application provides a food comprising the above composition. For example, a food comprising crushed Zanthoxylum or Connarus plant body, an extract obtained by extracting Zanthoxylum or Connarus plant body with a solvent containing water, or a concentrate or dried product of this extract may be included. In one embodiment, the food of the present application is a crushed Zanthoxylum or Connarus plant body, or a food comprising the same. Generally, Japanese pepper is used in a variety of ways as a spice, so it is conceivable to mix the composition of the present application into seasonings, furikake, spice mixes, curry powder or curry roux, tsukudani, etc. Furthermore, like spices such as ginger or herbs, it is conceivable to mix the composition into bread, confectionery, soft drinks, alcoholic beverages, etc.

In one embodiment, the food of the present application is an extract obtained by extracting the plant body of Zanthoxylum japonicum or Zanthoxylum connarus with a solvent containing water, or a food containing the same. An example of an extract obtained by extracting the plant body of Zanthoxylum japonicum or Zanthoxylum connarus with a solvent containing water is a beverage similar to tea, which uses hot water as a solvent. Therefore, a processed product in which crushed plant body of Zanthoxylum japonicum or Zanthoxylum connarus is packed in a pack or the like to facilitate hot water extraction also falls under the food containing the composition of the present application, and the hot water extract also falls under the food of the present application. In one embodiment, the food of the present application is a concentrate or dried product of an extract of Zanthoxylum japonicum or Zanthoxylum connarus, or a food containing the same. The concentration or drying may be for the purpose of improving the convenience of storage and transportation, and the concentrated or dried food may be diluted or reconstituted with water before being ingested, or may be mixed with other foods before being ingested. In one embodiment, the extract may be obtained by adding an excipient such as dextran or dextrin, and then drying the extract by a vacuum drying method, a freeze drying method, a spray drying method, or the like, and then drying the extract to a powder. The powder obtained by such a drying method may be dissolved in water again. In one embodiment, crushed Zanthoxylum or Zanthoxylum conulata plant bodies, an extract obtained by extracting Zanthoxylum or Zanthoxylum conulata plant bodies with a solvent containing water, or a concentrate or dried product of this extract may be steamed, and the resulting steam may be ingested.

Various foods can be used as food ingredients to be combined. For example, seafood, meat, vegetables, fruits, seasonings, and beverages. Not only fresh foods but also pre-processed foods can be used. Here, seafood can be not only fish meat but also fish eggs. Examples of meat include beef, pork, and chicken. Vegetables and fruits are mainly the parts to be eaten (e.g., fruit flesh), but may also include the peel. Examples of seasonings include soy sauce, miso, and sauce. Examples of beverages include coffee, black tea, green tea, soft drinks, fruit drinks, vegetable drinks, and dairy drinks. Not only a single ingredient but also multiple ingredients may be used as the food ingredient of the present invention. In addition, additives such as salt, sugar, acidulant, flavoring, coloring, various vitamins and minerals, protein, and dietary fiber may be added as necessary.

It is preferable to subject the food raw material to the necessary pretreatment in advance so that it is easy to mix the composition of the present application with the food raw material to be combined. For example, if the food raw material used is a vegetable, the pretreatment here includes preparatory work such as removing parts that are not suitable for eating, such as the skin or seeds, shredding to an appropriate size, and heating in advance by steaming, boiling, etc. Furthermore, if the food raw material used is meat, the pretreatment here also includes work of processing the meat into minced meat, etc.

In one embodiment, the components derived from the plant body of Zanthoxylum japonicum or Connarus japonicum can be prepared and provided in any dosage form, such as powder, granules, or liquid, in accordance with conventional methods using pharma- ceutically acceptable carriers such as dextran, dextrin, or cyclodextrin, or any other auxiliary agent, and can be used by blending with other compositions, or as tablets, capsules, granules, pills, powders, lozenges, liquids, elixirs, suspensions/emulsions, spirits, syrups, aromatic perfumes, or lemonades. In this case, examples of auxiliary agents that can be used include excipients, binders, disintegrants, lubricants, stabilizers, and odorants.

(Manufacturing method)
In one aspect, the present application provides a method for producing the composition described herein. In one embodiment, the method may include a step of crushing the plant body of Zanthoxylum japonicum or Connarus japonicum. In one embodiment, the plant body of Zanthoxylum japonicum or Connarus japonicum may be dried and then crushed, crushed and then dried, or a combination of these. Drying may be performed in the sun, using a dryer, or by freeze-drying. Known drying methods include natural drying, heat drying, water-absorbent drying, airflow drying, reduced pressure drying, vacuum drying, freeze-drying, high-frequency drying, air-conditioning drying, freeze-drying, and spray drying. In addition, the plant body may be crushed after pretreatment such as degreasing with a non-polar solvent such as hexane to obtain a component derived from the plant body. Crushing methods are known to those skilled in the art, and include methods using a ball mill, a roller mill, a hammer mill, a mortar and pestle, and a jet mill.

The crushed material obtained may be classified by sieving using particle size, or by classification using the difference in the distance traveled by wind or centrifugal force, to remove foreign matter and adjust the particle size. Classification methods are known to those skilled in the art and are broadly divided into methods using gravity or centrifugal force and methods using sieving, and examples of classification methods include those using a gravity classifier, a centrifugal classifier, a low-tap sieve shaker, an electromagnetic sieve shaker, or a sonic sieve shaker. In addition, in the crushing process, only a specific part of the plant body may be selected and crushed, or only a specific part may be selected after crushing. If necessary, the components derived from the plant of the present application may be mixed, kneaded, or stirred with other components. Methods of mixing, kneading, and stirring are known to those skilled in the art, and are broadly classified into a diffusion type in which the contents are mixed by rotating a container, and a convection type in which the contents are mixed by rotating a blade or screw, and include methods using a V-type mixer, a rotary oscillating mixer, a screw mixer, a ribbon mixer, and a paddle mixer. In one embodiment, the composition of the present application may contain moisture before or after crushing, and the moisture content is preferably 15% by weight or less, more preferably 10% by weight or less, and even more preferably 5% by weight or less. In one embodiment, additives such as a binder and a disintegrant may be added to the obtained powder to form granules. Methods for preparing granules are known to those skilled in the art, and include wet granulation methods (extrusion granulation method, fluidized bed granulation method, spray granulation method, rolling granulation method, stirring granulation method, and crushing granulation method), and dry granulation method (crushing granulation method).

In one embodiment, the present application provides a method for producing the above composition, comprising the step of extracting the plant body of Zanthoxylum japonicum or Connarus japonicum with a solvent containing water. As described above, it is preferable to use a polar solvent as the extraction solvent, for example, water, an aqueous solvent, and a hydrophilic organic solvent, and it is preferable to extract using one or more of these in combination at room temperature or at a temperature below the boiling point of the solvent. In one embodiment, a polar solvent may be used as the extraction solvent, and examples of hydrophilic organic solvents that can be used as the extraction solvent include lower aliphatic alcohols having 1 to 5 carbon atoms, such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones, such as acetone and methyl ethyl ketone; and polyhydric alcohols having 2 to 5 carbon atoms, such as 1,3-butylene glycol, propylene glycol, and glycerin.

When a mixture of two or more of the above polar solvents is used as the extraction solvent, the mixing ratio can be adjusted as appropriate. For example, when using a mixture of water and a hydrophilic organic solvent, a lower aliphatic alcohol, it is preferable to mix 1 to 90 parts by mass of the lower aliphatic alcohol with 10 parts by mass of water, when using a mixture of water and a lower aliphatic ketone, it is preferable to mix 1 to 40 parts by mass of the lower aliphatic ketone with 10 parts by mass of water, and when using a mixture of water and a polyhydric alcohol, it is preferable to mix 1 to 90 parts by mass of the polyhydric alcohol with 10 parts by mass of water.

The extraction process is not particularly limited as long as the soluble components contained in the extraction raw material can be dissolved in the extraction solvent, and can be carried out according to a conventional method. For example, the extraction raw material can be immersed in an extraction solvent in an amount 5 to 100 times (by mass) the amount of the extraction raw material, and the soluble components can be extracted at room temperature, under reflux heating, or under supercritical or subcritical conditions, and then the extraction residue can be removed by filtration to obtain an extract.

In one embodiment, the present application provides a method for producing the above-mentioned composition, comprising a step of concentrating or drying an extract obtained by extracting a plant body of Zanthoxylum or Connarus with a solvent containing water. Concentration methods are known to those skilled in the art, and can be carried out, for example, by removing impurities by activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, etc. Ion exchange resin treatment is particularly preferred. In addition, the solvent may be reduced by heating concentration, vacuum concentration, spray concentration, ventilation concentration, or freeze concentration. The concentrate obtained by such a concentration method may be diluted again with water. In one embodiment, the obtained extract may be dried. Drying methods are known to those skilled in the art, and include natural drying, heating drying, water absorbent drying, airflow drying, vacuum drying, vacuum drying, freeze drying, high frequency drying, air conditioning drying, freeze drying, and spray drying. An example of a drying method is a method in which an excipient such as dextran or dextrin is added to the extract, and then the extract is dried by a vacuum drying method, a freeze drying method, a spray drying method, or the like. The dried product obtained by such a drying method may be dissolved in water again.

[Example 1: Examination of the effect of grape pepper extract in preventing disuse syndrome]
(the purpose)
The following experiment was carried out to investigate the contribution of Japanese pepper extract to the prevention of disuse syndrome in the elderly. In particular, the efficacy of the non-edible part of Japanese pepper, which has not been studied much, was also examined.

(method)
Sample preparation As samples of Japanese pepper used in the experiment, fruits, mature leaves, inner bark, and seeds of Budou-Zanthyo were prepared. Figure 1 is a cross-sectional view of a Japanese pepper tree, showing the inner bark of the Japanese pepper. Mr. Takemasa Nitta kindly provided us with immature fruits (harvested in May), mature leaves (harvested in September), inner bark (harvested in September), and seeds (harvested in August) of Budou-Zanthyo produced in Wakayama Prefecture. Each Japanese pepper sample was freeze-dried immediately after harvest (freeze dryer FDU-2200 type: Tokyo Rikakikai Co., Ltd., Tokyo) and used in the experiment. Figure 2 shows each Japanese pepper sample after freeze-drying. Of each Japanese pepper sample, the immature fruit or inner bark sample was crushed with a mill mixer (FM-50: Sun Co., Ltd.), weighed out 5.0 g, mixed with 100 ml of tap water, and heated at 80°C or higher for 30 minutes to obtain a hot water extract. These hot water extracts were then diluted 10 times with tap water to obtain sample extracts.

Disuse syndrome model rats As shown in Figure 3, the rats were suspended by the tail using a tail suspension clip for disuse muscle atrophy research (PTC2010 type: Yamashita Giken Co., Ltd., Tokushima) with only the front legs touching the floor. The hind legs were not touching the floor, and the rats were raised with only the front legs, inducing disuse syndrome of the hind legs. The model was created and the animals were raised under the guidance and supervision of a veterinarian.

Breeding: 25 male Wistar rats aged 9 weeks were purchased from CLEA Japan, and were allowed to freely consume commercial solid feed CE-2 (CLEA Japan, Osaka) and tap water for one week as an acclimation period. After the acclimation period, the rats were divided into four groups from the age of 10 weeks, including a non-hindlimb suspension group that consumed water (water-hindlimb suspension (-): normal breeding, 6 rats), a hindlimb suspension group that consumed water (water-hindlimb suspension (+): positive control group for disuse syndrome, 6 rats), a hindlimb suspension group that consumed hot water extract of immature fruit (fruit-hindlimb suspension (+): 6 rats), and a hindlimb suspension group that consumed hot water extract of inner bark (inner bark-hindlimb suspension (+): 7 rats). The rats were given CE-2 feed and were allowed to freely consume tap water or each hot water extract as a drink. The three hindlimb-suspended groups were housed in large plastic cages equipped with hindlimb suspension devices, while the non-hindlimb-suspended groups were housed in similarly sized large plastic cages. The animals were housed individually for 10 days. During the housekeeping period, the animals were weighed once a week and their water intake was measured every other day. The housekeeping conditions were a room temperature of 22±2°C, humidity of 40-60°C, and lights on from 7:00 a.m. to 7:00 p.m. The animals were housed by a specialist and approved by the Animal Experiment Management Committee of Katagiri Veterinarian's Office (approval number: KWU19-01).

Dissection and Measurement Items After the breeding period was completed, whole blood was collected from the abdominal aorta under triple anesthesia, and major organs were observed and autopsied. Blood was centrifuged to obtain serum, and albumin (ALB), creatine phosphokinase (CPK), urea nitrogen (BUN), and alanine aminotransferase (ALT) were measured using an animal blood chemistry automatic analyzer (Fuji Dry Chem 3500V: Fuji Film Co., Ltd., Tokyo). The soleus muscle was extracted, weighed, and then immediately fixed, stained with Hematoxilin-Eosin (HE), and the maximum muscle circumference and maximum muscle cross-sectional area were calculated. In addition, after weighing the femur, the bone length was measured using a digital caliper (Digi Canon: Nakamura Seisakusho Co., Ltd., Tokyo), and bone strength (energy required to break bone (mJ), maximum bending load (N)) was measured using a bone strength tester (TK-252D: Muromachi Kikai Co., Ltd., Kyoto).

Staining After collecting the soleus muscle from the right side of each rat, it was immediately fixed in 10% neutral buffered formalin solution and entrusted to Kyodo Pathology Co., Ltd. to prepare paraffin blocks. The prepared blocks were thinly sliced, scooped with slide glass, and dried to prepare sections. The sections were immersed in xylene I, xylene II, xylene III, 100% ethanol I, 100% ethanol II, 100% ethanol III, 90% ethanol, and 70% ethanol, in that order, for 3 minutes each, and deparaffinized. The sections were washed with running water for 3 minutes and stained with hematoxylin solution for 10 minutes. The sections were washed with running water for about 5 minutes and stained with eosin solution for 10 minutes. The sections were immersed in 70% ethanol, 80% ethanol, 90% ethanol, 100% ethanol I, 100% ethanol II, and 100% ethanol III, in that order, for 3 minutes each, and then immersed in xylene I, xylene II, and xylene III, for 3 minutes each. This was observed using a microscope digital system, Moticam 2 (Shimadzu Rika Co., Ltd., Tokyo), and the maximum muscle circumference and maximum muscle cross-sectional area were calculated.

Statistical Processing The results were shown as the mean ± standard deviation, and the difference in the mean values was examined using multiple comparison (Tukey's test) with a significance level of 5%, using statistical analysis software IBM SPSS Statistics 19 (SPSS).

(result)
Effect on the physical condition of rats First, the effect of the experiment on the physical condition of the rats was examined. Figures 4 and 5 show the change in body weight and water intake of the rats during the experimental period, respectively. The body weight of the rats increased during the experimental period, but compared with the normally raised non-hindlimb-suspended group, the three hindlimb-suspended groups showed a tendency for the weight gain to be suppressed, and this was significant in the fruit-hindlimb-suspended (+) group and the inner bark-hindlimb-suspended (+) group. In addition, there was no change in water intake before and after suspension in any group. Table 1 shows the weight of the liver and kidney, and there was no change among the four groups. Table 2 shows the blood biochemistry properties (CPK: creatine phosphokinase, ALB: albumin, BUN: urea nitrogen, and ALT: alanine aminotransferase), and there was no change in any of the four items in the four groups. ALB is known as an index of eating disorder, and it was difficult to measure the amount of food intake in the cage used in this study, but since there was no difference in ALB, it was indicated that there was no difference in the amount of food intake. Furthermore, BUN and ALT are indicators of liver and kidney function, and since there were no differences in these, it was shown that there was no effect on liver function or kidney function.

Effect on disuse bone atrophy Next, the effect of ingestion of Japanese pepper hot water extract on disuse bone atrophy was examined. Table 3 shows the femoral bone survey items (bone weight, bone length, energy required to break bone, and maximum bending load). Compared with the normally raised water-hindlimb suspension (-) group, the water-hindlimb suspension (+) group, which is a positive control for disuse syndrome, had a significant decrease in bone weight, and the other hindlimb suspension groups also had a similar decrease in bone weight, indicating the symptoms of disuse bone atrophy due to hindlimb suspension. However, in the fruit-hindlimb suspension (+) group, which was given the hot water extract of the fruit while being suspended from the hindlimb, the energy required to break the bone was significantly increased compared to the water-hindlimb suspension (+) group, and was comparable to the normal water-hindlimb suspension (-) group (water-hindlimb suspension (-) group, 63.8±7.4mJ; water-hindlimb suspension (+) group, 38.4±11.7mJ; fruit-hindlimb suspension (+) group, 60.3±16.7mJ). In addition, the inner bark-hindlimb suspension (+) group, which was given the hot water extract of the inner bark, also showed a tendency to increase the energy required to break the bone compared to the water-hindlimb suspension (+) group (49.9±16.3mJ). This indicates that the ingestion of the hot water extract of the fruit suppressed the occurrence of disuse bone atrophy. A similar tendency was also observed in the item of maximum bending load (stiffness). In other words, the group that ingested the hot water extract of Japanese pepper had an increased maximum bending load compared to the group that ingested only water, and the value was close to the maximum bending load in healthy rats. These results indicate that ingesting the hot water extract of Japanese pepper inhibits disuse bone atrophy.

Effect on disuse muscle atrophy The effect of ingestion of Japanese pepper hot water extract on disuse muscle atrophy was investigated. Figure 6 shows the weight of the soleus muscle in each group. Compared with the normally reared water-hind limb suspension (-) group, the muscle mass was reduced in the hind limb suspension (+) group, indicating symptoms of disuse muscle atrophy due to hind limb suspension. However, there was a tendency for the reduction in muscle mass to be suppressed by ingestion of Japanese pepper hot water extract. Figure 7 shows an image of the HE-stained soleus muscle cross section, and Table 4 shows the maximum muscle circumference and maximum muscle cross-sectional area of the soleus muscle. Again, compared with the normally reared water-hind limb suspension (-) group, atrophy of muscle fibers was confirmed in the 3 groups of the hind limb suspension (+) group, but there was a tendency for muscle atrophy to be suppressed by ingestion of Japanese pepper hot water extract.

(Consideration)
Most research on Sansho has been limited to sanshool, the compound that gives it its characteristic aroma and pungent flavor, and its derivatives.

When it is used as a food product, it is preferable to use an extract method that does not use organic solvents, etc., and to make it in the form of an easy-to-ingest beverage. Therefore, the preventive effect against disuse syndrome was investigated using a hot water extract of Japanese pepper. However, the preventive effect against disuse syndrome observed in this specification is not limited to hot water extracts of Japanese pepper, and is expected to be observed in any substance derived from Japanese pepper. In addition to the immature fruit that is generally used for food, the inner bark, which is a non-edible part, was selected, and hot water extracts of these were administered to hindlimb-suspended rats, a model of disuse syndrome. The results of blood chemistry analysis, body weight measurement, and water intake measurement showed that the amount of food and water intake did not change in each rat group, and body weight also increased. There was no change in liver function or kidney function, and no adverse effect on physical condition was observed due to the intake of hot water extract of Japanese pepper.

When femur characteristics were examined, bone mass was reduced in the hindlimb suspension group compared to the normal group, but the energy required for bone fracture and maximum bending load were reduced to a smaller extent in the group that had been fed Japanese pepper hot water extract, indicating the inhibitory effect of Japanese pepper hot water extract on disuse bone atrophy. Next, when soleus muscle characteristics were examined, muscle mass, maximum circumference, and maximum cross-sectional area were reduced in the hindlimb suspension group compared to the normal group, but the extent of reduction tended to be smaller in the group that had been fed Japanese pepper hot water extract, suggesting the inhibitory effect of Japanese pepper hot water extract on disuse muscle atrophy. Muscle atrophy is also observed when energy intake is reduced or protein catabolism is difficult, but since there was no difference in blood albumin concentration (ALB) between the groups, it is indicated that the muscle atrophy shown is due to immobility.

In this experiment, it was demonstrated that hot water extract of Japanese pepper has the effect of inhibiting disuse bone atrophy. The results of this study will lead to new uses for Japanese pepper.

[Example 2: Effects of hot water extract of Japanese pepper fruit and hot water extract of Connarus on soleus muscle and femur strength in disuse syndrome model rats]
(the purpose)
The above Example 1 showed that the hot water extract of Budou Sansho suppressed the decrease in femoral strength in rats with artificially induced disuse syndrome. In this Example, experiments were carried out using Asakura Sansho, a Japanese pepper of the same genus, and Connarus, a plant of the Mammalian family.

(method)
Disuse syndrome model rats Twenty-one 9-week-old male Wistar rats were purchased from Kiwa Animal Experiment Institute Co., Ltd. and were acclimated for one week. After acclimation, the rats were classified into four hindlimb suspension groups and one non-hindlimb suspension group. The hindlimb suspension groups were further classified into the water-hindlimb suspension (+) group (n=3, also denoted as Cont(+)) administered with tap water, the grape-hindlimb suspension (+) group (n=3, also denoted as Grape(+)) administered with Budo Sansho hot water extract, the Asakura-hindlimb suspension (+) group (n=6, also denoted as Asakura(+)) administered with Asakura Sansho hot water extract, and the Conalus-hindlimb suspension (+) group (n=6, also denoted as Conalus(+)) administered with Conalus hot water extract (n=3, 3, 6, and 6, respectively). The non-hindlimb-suspended group, in which hindlimb suspension was not performed, was administered tap water and designated as the water-hindlimb-suspended (-) group (n=3; also referred to as Cont(-)).

Breeding The animals were fed a commercially available solid feed CE-2 (CLEA Japan, Osaka), and were given free access to feed and drinking water. After habituation, the hindlimb-suspended group was suspended by the tail using a tail suspension clip for disuse muscle atrophy research (RTC2010 type: Yamashita Giken Co., Ltd., Tokushima). The animals were bred with only the forelimbs, without the hindlimb touching the floor, to induce disuse muscle atrophy in the hindlimb. Model creation and animal breeding were carried out under the guidance and supervision of a veterinarian. Both the hindlimb-suspended group and the non-hindlimb-suspended group were bred in large plastic cages, and were individually bred for about 10 to 12 days according to a previous report. During the breeding period, the amount of water consumed was measured every other day, and the body weight was measured once a week. The animals were bred in a breeding room with a room temperature of 22±2°C and lights on from 7:00 a.m. to 7:00 p.m. This experiment was carried out with the approval of the Animal Experiment Management Committee of Katagiri Veterinarian's Office.

Sample preparation Grape sansho fruits from Wakayama Prefecture were provided by a partner farm. Asakura sansho from Hyogo Prefecture was purchased directly from the farm. Each sample was completely dried in a vacuum at minus 80 degrees or less using a freeze dryer (FDU-2200 type: Tokyo Rikakikai Co., Ltd.) and stored in a desiccator until the hot water extract was prepared. The inner bark of the original Conalus rube from Brazil was used as the Conalus. The hot water extract to be administered to the animals was prepared immediately before administration. That is, each sample was crushed in a mill mixer (FM-50: Sun Co., Ltd., Tokyo), 5.00 g was weighed out, mixed with 100 ml of tap water, and heated to 80 degrees or more for 30 minutes. The hot water extract was diluted 10 times with tap water and administered.

Dissection and Measurement Items After the breeding period, whole blood was collected from the abdominal aorta under triple anesthesia, and major organs were observed and autopsy was performed. The soleus muscle was removed and weighed, then immediately fixed in 10% neutral buffered formalin and stained with HE (Hematoxylin-eosin). The femur was removed and stripped of muscle tissue, and the bone length was measured using a digital caliper (Digi-Canon: Nakamura Seisakusho Co., Ltd., Tokyo). The maximum bending load (N) and the energy required to break the bone (mJ) were measured using a bone strength tester (TK-252D: Muromachi Kikai Co., Ltd., Kyoto). The measurement was performed by displacing the plunger from above at a speed of 10 mm/min at the midpoint between the two supports, with the distance between both ends being constant.

Statistical processing The results are shown as the mean ± standard deviation, and the difference between the mean values was examined using the statistical analysis software IBMSPSS Statistics23 (SPSS, USA) and confirmed by multiple comparison (Tukey-Kremar). The risk level was set at 5%.

(result)
Induction of disuse syndrome No difference was observed between the body weights of each group at the time of necropsy at the end of the breeding period (Table 5, left). The weight of the soleus muscle extracted after the whole blood collection was significantly lower in all four hindlimb suspension groups compared to the water-hindlimb suspension (-) group, which was a non-hindlimb suspension group, indicating that disuse muscle atrophy was induced by hindlimb suspension (Table 5, center). In addition, there was no significant difference in femur weight between the hindlimb suspension groups and the water-hindlimb suspension (-) group (Table 5, right).

Effects on disuse bone atrophy The results of the bone fracture test using the excised femurs are shown in Figure 8. The maximum bending load (N), which indicates the degree of bone flexibility, was significantly lower in each of the four hindlimb suspension groups compared to the non-hindlimb suspension group, water-hindlimb suspension (-), suggesting that the decrease in maximum bending load may be due to the effect of hindlimb suspension. In addition, among the four hindlimb suspension groups, the maximum bending load was significantly higher in the grape-hindlimb suspension (+) group, the Asakura-hindlimb suspension (+) group, and the Conarus-hindlimb suspension (+) group compared to the water-hindlimb suspension (+) group. This suggests that the degree of bone flexibility may have been improved by administration of the Budo Sansho hot water extract, the Asakura Sansho hot water extract, or the Conarus hot water extract.

(Consideration)
This experiment was conducted as a follow-up study on the muscle atrophy prevention effect of Budo Sansho conducted in Example 1. Looking at the results of this experiment alone, the group administered Asakura Sansho hot water extract showed a similar tendency to the group administered Budo Sansho hot water extract. The preventive effect of disuse syndrome was observed in both Zanthoxylum species, suggesting that the preventive effect of disuse syndrome is a property shared by all Zanthoxylum species.

In addition, when bone strength was measured by bone fracture, the maximum bending load (N), which indicates the degree of bone flexibility, was significantly higher in the grape-hindlimb suspension (+) group, Asakura-hindlimb suspension (+) group, and Conarus-hindlimb suspension (+) group compared to the water-hindlimb suspension (+) group. This result indicates that not only sansho (especially Budo sansho and Asakura sansho), but also Conarus has a preventive effect on disuse bone atrophy caused by hindlimb suspension.

Claims (8)

A composition containing an ingredient derived from the Connarus plant for preventing or treating disuse syndrome, wherein the plant contains the inner bark, the disuse syndrome includes disuse bone atrophy, and the ingredient comprises crushed material of the plant, or an extract obtained by extracting the crushed material with a solvent containing water, or a concentrate or dried product of the extract . The composition of claim 1, wherein the plant further comprises fruits, mature leaves, and/or seeds. The composition of claim 1 or 2, wherein the ingredients include ground Connarus plant matter. The composition according to claim 1 or 2 , wherein the component comprises an extract obtained by extracting the crushed material with a solvent containing water. The composition according to claim 1 or 2 , wherein the ingredient comprises a concentrate or a dried product of an extract obtained by extracting the crushed material with a solvent containing water. The composition according to any one of claims 1 to 5, which is intended for oral intake. A food for preventing or treating disuse syndrome, comprising the composition according to any one of claims 1 to 6, wherein the disuse syndrome includes disuse bone atrophy. A method for producing the composition according to any one of claims 1 to 6 or the food according to claim 7, comprising the steps of:
A manufacturing method comprising a step of extracting crushed material of the Conalus plant with a solvent containing water.