US20170182109A1 - Appetite-suppressing agent - Google Patents

Appetite-suppressing agent Download PDF

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US20170182109A1
US20170182109A1 US15/455,586 US201715455586A US2017182109A1 US 20170182109 A1 US20170182109 A1 US 20170182109A1 US 201715455586 A US201715455586 A US 201715455586A US 2017182109 A1 US2017182109 A1 US 2017182109A1
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appetite
component
liquid component
molecular weight
indian mulberry
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Masatomo Mori
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Yada Toshihiko
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Masatomo Mori
Toshihiko Yada
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Assigned to MORI, MASATOMO, YADA, Toshihiko reassignment MORI, MASATOMO ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: MORI, MASATOMO
Assigned to MORI, MASATOMO reassignment MORI, MASATOMO ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: YADA, Toshihiko
Publication of US20170182109A1 publication Critical patent/US20170182109A1/en
Priority to US16/451,214 priority Critical patent/US11260098B2/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/74—Rubiaceae (Madder family)
    • A61K36/746—Morinda
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/30—Dietetic or nutritional methods, e.g. for losing weight
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • the present findings show an appetite-suppressing composition. More specifically, the present invention is associated with an appetite-suppressing composition derived from a specific plant•fruit.
  • Obesity is defined by overweight, which is obviously associated with high incidences of lifestyle related disorders such as hyperglycemia, hypertension and hyperlipidemia.
  • Japan Society for the Study of Obesity states that obesity is defined by 25 or more of body mass index (BMI), and that obesity disease is diagnosed by the case, in which obesity shows at least one health problem including hyperglycemia, hypertension, hyperlipidemia, fatty liver, a sleep apnea syndrome, hyperuricemia, a coronary artery disorder, a cerebrovascular disorder, a menstrual disorder, an orthopedic disorder, and obesity-related nephropathy.
  • Obesity disease is also diagnosed by the other case, in which obesity shows a certain value or more of the visceral fat amount (Non-Patent Document 1).
  • non-Patent Document 1 Furthermore, on the basis of obesity due to an increase in visceral fat, metabolic syndrome is diagnosed and significantly associated with cerebrovascular and cardiovascular disorders. Therefore, it is important that obesity disease and metabolic syndrome should be treated prior to development
  • Body weight is maintained at a constant level by the mechanism of energy homeostasis, to which appetite and energy expenditure mainly contribute.
  • Obesity is caused by dysregulation of energy homeostasis, i.e., increased appetite (overeating) over a long-period of time than energy expenditure.
  • Appetite is mainly regulated by the hypothalamus in the brain, and it has recently become known that appetite is substantially regulated by active molecules, which are synthesized and secreted from peripheral adipocytes and affect on the brain hypothalamus.
  • Non-Patent Document 2 nesfatin-1 suppressed differentiation and proliferation of adipocytes
  • the approximately 30%-components are known to be derived from certain kinds of plants.
  • a component showing the same activity as nesfatin-1 contained in a plant and/or fruit which may be commercially available as a food stuff or drinking material, its component is expected to show appetite suppression, resulting in improvement of obesity, from the aforementioned point of view.
  • a liquid component extracted from plants and fruits shows the appetite-suppressing activity such as nesfatin-1.
  • Non-Patent Document 1 Contemporary clinical obesity, Summary of criteria for diagnosis of obesity: Nippon Rinsho 72: 13-18, 2014.
  • Non-Patent Document 2 Identification of nesfatin-1 as a satiety molecule in the hypothalamus: Nature 443: 709-712, 2006.
  • Non-Patent Document 3 Morinda citrifolia L. (Noni): a review of the scientific validation for its nutritional and therapeutic properties. Journal Diabetes and Endocrinology 3: 77-91, 2012.
  • the present invention is devised in view of the current states of the aforementioned techniques of a related art, and it is devised to provide an appetite-suppressing composition derived from a specific plant fruit and to determine the effect of the composition.
  • the present invention devised to achieve the aforementioned purpose is related to the following [1] to [14].
  • An appetite-suppressing composition characterized by containing a component derived from Indian mulberry ( Morinda citrifolia ) as an effective component.
  • An appetite-suppressing composition characterized by being a liquid component derived from a fruit of Indian mulberry ( Morinda citrifolia ).
  • a food product which consists of or contains the appetite-suppressing composition as described in any one of [1] to [11].
  • the present invention provides the appetite-suppressing composition with excellent effects that possess and/or consists of an effective liquid component derived from, for example, a fruit of Indian mulberry.
  • the administration of the appetite-suppressing composition, as shown in the present invention demonstrates a significant reduction of food intake, resulting in prevention and amelioration of obesity and health problems involving obesity disease and metabolic syndrome. Furthermore, the intake of the appetite-suppressing composition, as shown in the present invention, more easily induces dieting of obese human subjects.
  • FIG. 1 shows the concentration dilution curve of the undiluted solution of liquid component of Indian mulberry, in which nesfatin-1 was detected by using the Nesfatin-1 ELISA kit.
  • FIG. 2 shows the body weight and the amounts of daily food intake before experiment as shown in FIGS. 3 and 4 .
  • FIG. 3 shows the changes in the cumulative amounts of food intake in mice 0.5 ⁇ 3 hours after the administration of the diluted and undiluted solution of liquid component of Indian mulberry.
  • FIG. 4 shows the changes in the cumulative amounts of food intake in mice 6 ⁇ 48 hours after the administration of the diluted and undiluted solution of liquid component of Indian mulberry.
  • FIG. 5 shows the body weight and the amounts of daily food intake before experiment as shown in FIGS. 6 and 7 .
  • FIG. 6 shows the changes in the cumulative amounts of food intake in mice 0.5 ⁇ 3 hours after the administration of the undiluted solution of liquid component of
  • Indian mulberry and the residual liquid containing the >3,000 component that was obtained using the molecular weight cut-off filter device were obtained using the molecular weight cut-off filter device.
  • FIG. 7 shows the changes in the cumulative amounts of food intake in mice 12 ⁇ 48 hours after the administration of the undiluted solution of liquid component of Indian mulberry and the residual liquid containing the >3,000 component.
  • FIG. 8 shows the body weight and the amounts of daily food intake before experiment as shown in FIGS. 9 and 10 .
  • FIG. 9 shows the changes in the cumulative amounts of food intake in mice 0.5 ⁇ 3 hours after the administration of the diluted and undiluted solution of the residual liquid containing the >3,000 component.
  • FIG. 10 shows the changes in the cumulative amounts of food intake in mice 12 ⁇ 24 hours after the administration of the diluted and undiluted solution of the residual liquid containing the >3,000 component.
  • FIG. 11 shows the body weight and the amounts of daily food intake before experiment as shown in FIGS. 12 and 13 .
  • FIG. 12 shows the changes in the cumulative amounts of food intake in mice 0.5 ⁇ 3 hours after the administration of the undiluted solution of liquid component of Indian mulberry and the filtrated liquid containing the ⁇ 3,000 component that was obtained using the molecular weight cut-off filter device.
  • FIG. 13 shows the changes in the cumulative amounts of food intake in mice 12 ⁇ 24 hours after the administration of the undiluted solution of liquid component and the filtrated liquid containing the ⁇ 3,000 component.
  • FIG. 14 shows the preference ratio for the intake of saccharine solution in the conditioned mice after the administration of saline, the undiluted solution of liquid component of Indian mulberry, the residual liquid containing the >3,000 component, the filtrated liquid containing the ⁇ 3,000 component, and lithium chloride.
  • Non-Patent Document 2 A novel molecule, which was found as a secretory protein showing expression in both brain hypothalamus and adipocytes, and activation by the peroxisome proliferator-activated receptor ⁇ , i.e., a master regulator of metabolism, was named as nesfatin by the inventors of the present invention (Non-Patent Document 2). According to processing of nesfatin, nesfatin-1 and nesfatin-2/3 were generated. Nesfatin-1 was present in rat spinal fluids and human blood. When nesfatin-1 was centrally administered to the rat ventricle, food intake was significantly suppressed, resulting in reduction of body weight.
  • nesfatin-1 In contrast, when the expression of nesfatin was decreased in the hypothalamus, food intake was elevated, leading to increases in body weight. Furthermore, the intracerebral administration of nesfatin-1 increased energy expenditure. On the other hand, nesfatin-2/3 did not exhibit the activity of appetite suppression or body weight reduction. Subsequent studies demonstrated that nesfatin suppressed proliferation of adipocytes and that the intraperitoneal administration of nesfatin-1 induced appetite suppression in a dose-dependent manner.
  • liquid component and/or drinking water derived from plants and/or fruits which are commercially available.
  • the ELISA kit with high sensitivity and high specificity for nesfatin-1, considerable efforts were devoted to detect nesfatin-1 using these liquid components.
  • the liquid component extracted from Indian mulberry (scientific name, Morinda citrifolia; general name, Indian mulberry) possessed nesfatin-1 and nesfatin-1-like molecules at a relatively high concentration. Accordingly, the undiluted solution of liquid component of Indian mulberry showed a significant suppression of food intake, whose suppression was not a non-specific effect.
  • Indian mulberry Morinda citrifolia
  • Morinda citrifolia is a small evergreen tree belonging to Rubiaceae, and it is known to be naturally found in part of Ryuku islands and Ogasawara islands in Japan.
  • a liquid component may be mentioned.
  • Preferred examples include a liquid component extracted from a fruit of Indian mulberry, in particular, an extract after fermenting the fruit for a suitable period, for example, 3 months or longer period.
  • the appetite-suppressing composition of the present invention shows, as an effective component, the aforementioned liquid component extracted from Indian mulberry, for example. It is also possible that the liquid component was prepared as a supernatant by means of centrifugation, or was fractionated to obtain the components showing the different molecular weights such as >3,000 daltons and ⁇ 3,000 daltons, using the molecular weight cut-off filter device. It is also possible to adjust the concentration of these components using saline and buffer solutions.
  • the examples as the forms and systems of administration, using the appetite-suppressing composition of the present invention include an oral administration (employing a tablet, a capsule, a granule, a powder, a syrup, an enteric-dissolved preparation, a troche, or a drink preparation), a parenteral preparation like an injection solution, a suppository, a transdermally-absorbed preparation, and a preparation for outer application.
  • an oral administration employing a tablet, a capsule, a granule, a powder, a syrup, an enteric-dissolved preparation, a troche, or a drink preparation
  • a parenteral preparation like an injection solution, a suppository, a transdermally-absorbed preparation, and a preparation for outer application.
  • those formulations are prepared by using only the appetite-suppressing component of the present invention, or by appropriately combining its component with a certain vehicle (for example, saccharides such as sorbitol, glucose, lactose, dextrin, or starch, inorganic materials such as calcium carbonate, crystalline cellulose, saline, distilled water, sesame oil, corn oil, olive oil, safflower oil, and mixture of amino acids), a binder, a smoothing agent, an extender, a disintegrant, a surface active agent, a lubricant, a dispersant, a suspending agent, an emulsifying agent, a buffering agent, a preservative, a flavor, a fragrance, a coating agent, a carrier, a diluent, and an anti-oxidizing agent.
  • a certain vehicle for example, saccharides such as sorbitol, glucose, lactose, dextrin, or starch, inorganic materials such as calcium
  • any form may be employed.
  • the preferred form is the oral administration using an enteric-dissolved preparation, and the doses of oral administration are not particularly limited as long as the effect obtained.
  • the amounts of intake•administration may vary depending on a health condition, body weight, sex, or age of subjects, and other factors.
  • the doses of administration are comparable to those of 600 mg to 3,600 mg (per kg of body weight) in terms of the amount of liquid component.
  • the appetite-suppressing composition can be taken or administered depending on any intake•administration schedule, it should be considered to be administered on several separated times per day, and its administration should be continued from several weeks to several months and more.
  • subjects taking the appetite-suppressing composition of the present invention are not particularly limited to subjects with obesity who require the composition.
  • preferred are subjects with obesity who exhibit obesity disease and metabolic syndrome, subjects with obesity who tend to show health problems associated with obesity, mammals other than human subject who desire prevention and amelioration of lifestyle-related disorders, and subjects under dieting and physical exercise therapy for body slimming.
  • the appetite-suppressing composition of the present invention having the constitution, as described above, clearly showed appetite suppression. Particularly, the administration of the residual liquid containing the >3,000 component induced the enhanced appetite suppression with a long-period of efficiency.
  • the appetite-suppressing composition of the present invention is also useful upon mixing with food products including a processed food, a health food and a health drink like a so-called food for specified health uses.
  • examples of materials mixed with the appetite-suppressing composition of the present invention include a processed wheat-flour product represented by bread and noodles; a processed rice product like rice gruel, rice cooked with seasoned ingredients; snacks like biscuit, cake, jelly, chocolate, senbei (rice cracker), and ice cream; a processed soybean product such as tofu and processed tofu product; drinks such as a soft drink, a fruit juice drink, a vegetable drink, a squeezed liquid of mixed fruits, a squeezed liquid of mixed vegetables; a milk drink, and a carbonate drink; a dairy product such as yoghurt, cheese, butter, or milk; a seasoning such as soy sauce, sauce, miso, mayonnaise, or dressing; a meat or a processed meat product such as ham, bacon, or sausage; a processed sea food product such as hanpen (floated-type kamaboko), chikuwa (fish stick), fish can; a seasoning; and a fry oil.
  • an oral•enteric nutrition food product or a functional food product such as a tablet food like capsule, a concentrated fluid food, a natural fluid food, a semi-digested nutrition food, an elemental nutrition food, or a drink nutrition food can be produced.
  • animal foods include a food for small animal like rabbit, rat, and mouse, and a pet food used for dog, cat, small bird, or squirrel.
  • Feeds and food products as various forms can be produced by suitably mixing the appetite-suppressing composition of the present invention with other food materials and/or substances including a solvent, a softening agent, an oil, an emulsifying agent, a preservative, a fragrance, a stabilizing agent, a coloring agent, an anti-oxidizing agent, a moisturizing agent, and a thickening agent.
  • Food stuff or drink as materials 22 kinds of liquids extracted from the plants•fruits as shown below, which are commercially available as food stuff and drink in public markets in the international street of Okinawa ken, were obtained. Each of liquid extracts was subjected to centrifuge at 1,500 rpm for 15 minutes to obtain a supernatant (this supernatant is described as the undiluted solution of liquid component). Twenty two materials were as follows;
  • nesfatin-1 was measured in each of the undiluted solutions of liquid component.
  • the concentrations of leptin and adiponectin, all of which are involved in the appetite regulation in the brain hypothalamus and are present in human blood and adipocytes were particularly measured by using an ELISA kit specific to each molecule, according to the methods manufactured by Mercodia.
  • ANOVA One-way analysis of variance
  • the constituents contained in the undiluted solution of liquid component obtained as mentioned in Method 1 were fractionated into two kinds of liquid components; one component, containing nesfatin-1, showing a molecular weight of 3,000 daltons or more (the same meaning is applied to >3,000 component), and the other component showing a low molecular weight of 3,000 daltons or less (the same meaning is applied to ⁇ 3,000 component).
  • the undiluted solution of liquid component was centrifuged at 3,600 rpm for 120 minutes using the above molecular weight cut-off filter device. The following centrifugation at 3,600 rpm for 60 minutes was carried out to fractionate and collect the filtrated liquid that was passed through the filter.
  • the protein concentrations were measured in the undiluted solution of liquid component and the residual liquid component containing the >3,000 component.
  • concentrations of potassium were measured by ion electrode selection method in the undiluted solution of liquid component, the residual liquid containing the >3,000 component, and the filtrated liquid containing the ⁇ 3,000 component.
  • concentrations of nesfatin-1 were measured by using the nesfatin-1 ELISA kit, as mentioned in Method 2.
  • amino acids in the undiluted solution of liquid component were measured by an automatic amino acid analyzing method.
  • Preference effect relating to conditioned taste aversion Appetite can be non-specifically suppressed by a certain kind of material, which induces vomiting, throwing-up, unpleasant feeling, or taste disorder.
  • a conditioned taste aversive test (a conditioned taste aversion preference test) was carried out in mice. For 5 days, each of mice was adapted to two bottles containing water, allowing to the drinking schedule of 2 hours per day (10:00 am to 12:00).
  • the administered concentration of each liquid component was a dose exhibiting approximately 1 ⁇ 2-food suppression of the saline-injected group as followed; a 1 ⁇ 4-dilution of the undiluted solution of liquid component obtained as mentioned in Method 1, a 1/16-dilution of the residual liquid containing the >3,000 component obtained as mentioned in Method 4, and the undiluted filtrated solution containing the ⁇ 3,000 component.
  • a lithium chloride solution was used at a dose of 3 nmo/kg of body weight.
  • the control was the group administered with saline.
  • the data are expressed as mean value ⁇ standard deviation. Determination of a significant difference among each group was carried out by One-way analysis of variance (ANOVA).
  • Detection 1 of nesfatin-1 in the undiluted solution of liquid component Many of the undiluted solutions of liquid component were used to detect nesfatin-1, and it was found for the first time that only the undiluted solution of liquid component of a fruit of Indian mulberry contained nesfatin-1 at relatively high concentrations. Another undiluted solutions of liquid component did not contain nesfatin-1.
  • the fruit of Indian mulberry was a product obtained by fermentation over 3 months approximately.
  • FIG. 1 shows the concentration dilution curve of the liquid component of a fruit of Indian mulberry, which was measured for nesfatin-1 using the nesfatin-1 ELISA kit.
  • the white column indicates the saline-injected group
  • the oblique-line column indicates the group administered with a 1/16-dilution
  • the striped column indicates the group administered with a 1 ⁇ 4-dilution
  • the black column indicates the group administered with the undiluted solution of liquid component.
  • * and ** Asterisks indicate significant differences (P ⁇ 0.05 and P ⁇ 0.01, respectively) compared with each of the comparable groups.
  • the residual liquid containing the >3,000 component which was obtained by fractionating the undiluted solution of liquid component of Indian mulberry using a molecular weight cut-off filter device, showed the protein concentrations of 189 to 219 mg/ml, whereas the undiluted solution of liquid component showed the protein concentrations of 50 to 58 mg/ml.
  • the potassium concentrations were 46.6 mg in the undiluted solution of liquid component and were 4.1 mg in the residual liquid containing the >3,000 component, showing that the residual ratio of potassium was 8.8% on average. Accordingly, the potassium concentrations in the filtrated liquid containing the ⁇ 3,000 component were 44.2 mg, showing that the filtrated ratio of potassium was calculated to be 94.8% on average.
  • the nesfatin-1 concentrations in the filtrated liquid containing the ⁇ 3,000 component ranged 0.5 to 10.3 ng/ml (average, 3.1 ng/ml), indicating that the filtrated concentration of nesfatin-1 was only 5.6% of the concentration of the undiluted solution of liquid component.
  • Amino acids in the undiluted solution of liquid component showing a concentration of 1 mg/ml over were as follows; glutamic acid with molecular weight 147 daltons (1.89 mg/ml), asparaginic acid with 133 daltons (1.37 mg/ml), arginine with 174 daltons (1.07 mg/ml), and alanine with 89 daltons (1.02 mg/ml).
  • the vertical axis represents the cumulative amounts of food intake (g) and the horizontal axis indicates the time (h) after administration.
  • the white column indicates the saline-injected group
  • the black column indicates the group administered with the undiluted solution of liquid component
  • the striped column indicates the group administered with the residual liquid containing the >3,000 component.
  • * and ** Asterisks indicate significant differences (p ⁇ 0.05 and P ⁇ 0.01. respectively) compared with the group administered with saline ( FIG. 6 ) and each of the comparable groups ( FIG. 7 ).
  • Appetite-suppressing effect-3 (enhancing activity): The residual liquid containing the >3,000 component obtained from the liquid component of Indian mulberry, and its 1 ⁇ 4- and 1/16-diluted solutions in saline were intraperitoneally administered to mice, and food intake of each of mice was measured. As shown in FIGS. 9 and 10 , the administration of the residual liquid containing the >3,000 component suppressed food intake in a dose-dependent manner. The concentration showing 1 ⁇ 2-suppression of food intake during 1 to 2 hours after administration was found to be a 1/16-dilution, indicating that the appetite-suppressing effect was approximately 4 times stronger than the undiluted solution of liquid component. In addition, the appetite-suppressing activity remained until 24 hours after administration.
  • FIGS. 9 and 10 The changes in the amount of food intake over time after administration are illustrated in FIGS. 9 and 10 .
  • the vertical axis represents the cumulative food intake (g) and the horizontal axis indicates the time (h) after administration.
  • the white column indicates the saline-injected group
  • the oblique-line column indicates the group administered with a 1/16-dilution
  • the striped column indicates the group administered with a 1 ⁇ 4-dilution
  • the black column indicates the group administered with the residual liquid containing the >3,000 component.
  • * and ** Asterisks indicate significant differences (P ⁇ 0.05 and P ⁇ 0.01, respectively) compared with the saline-injected group.
  • Appetite-suppressing effect-4 (component with low molecular weight):
  • the group administered with the filtrated liquid containing the ⁇ 3,000 component obtained from the liquid component of Indian mulberry exhibited significant appetite suppression as illustrated in FIGS. 12 and 13 . Its suppressing activity was nearly equivalent to the activity observed in the group administered with the un diluted solution of liquid component.
  • the filtered liquids contained components with low molecular weight such as potassium and amino acids. However, as illustrated in FIG. 13 , the activity of appetite suppression induced by the filtrated liquid containing the ⁇ 3,000 component was not observed after 24 hours of administration, indicating the activity similar to the undiluted solution of liquid component.
  • the body weight (BW) and 24 hour-food intake before experiment are illustrated in FIG. 11 .
  • the changes in food intake over time after administration are illustrated in FIGS. 12 and 13 .
  • the vertical axis represents the cumulative amounts of food intake (g) and the horizontal axis indicates the time (h) after administration.
  • the white column indicates the saline-injected group
  • the black column indicates the group administered with the undiluted solution of liquid component
  • the striped column indicates the group administered with the filtrated liquid containing the ⁇ 3,000 component.
  • * and ** Asterisks indicate significant differences (P ⁇ 0.05 and P ⁇ 0.01, respectively) compared with the saline-injected group.
  • the white column indicates the saline-injected group (A)
  • the black column indicates the group administered with the undiluted solution of liquid component (B)
  • the oblique-line column indicates the group administered with the residual liquid containing the >3,000 component (C)
  • the striped column indicates the group administered with the filtrated liquid containing the ⁇ 3,000 component (D)
  • the grey column indicates the group administered with lithium chloride (E). ** Asterisk indicates a significant difference (P ⁇ 0.01) compared with the saline-injected group.
  • the filtrated liquid containing the ⁇ 3,000 component which was prepared by means of the device of molecular weight cut-off filter, showed the significant appetite suppression, this filtrated liquid did not show any enhancing effect or long-term acting effect in suppressing appetite.
  • the present findings showed for the first time that the filtrated liquid containing the ⁇ 3,000 component, such as several amino acids derived from a fruit of Indian mulberry, possessed a appetite-suppressing effect without showing a non-specific activity.
  • the present invention is very excellent.

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