JP2022097521A - Anti-obesity potential of garcinol - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide means for the therapeutic management of obesity, and means for the modification of gut microbiota.

SOLUTION: Disclosed are compositions containing garcinol for the therapeutic management of obesity. More specifically, the invention relates to the use of garcinol for a) maintaining energy balance in mammalian adipose cellular systems, b) managing hypercholesterolemia and c) reducing weight gain in mammals. The modification of gut microbiota and the increase of beneficial microbe, Akkermansia muciniphila by garcinol are also disclosed.

SELECTED DRAWING: Figure 1b

COPYRIGHT: (C)2022,JPO&INPIT

Description

Mutual Reference of Related Patent Applications The present invention claims the priority of US Provisional Patent Application No. 625199949 filed June 15, 2017 and US Provisional Patent Application No. 62523611 filed June 22, 2017. It is a PCT application to be filed.
The present invention generally relates to compositions for weight management. Specifically, the present invention relates to compositions containing garcinol for the control of obesity, hypercholesterolemia, and alteration of the gut microbiota.

Obesity is considered to be a major health risk in the development of various disorders such as hypertension, type 2 diabetes, heart disease, stroke, osteoarthritis, and psychiatric disorders. Worldwide, more than 1 in 10 people are obese, and in the United States, about 36% of adults are obese (https://www.medicalnewstorey.com/articles/319902.php, May 10, 2018). Access on the day). Obesity results from an imbalance between the energy content of food eaten and the energy consumed by the body to sustain life and to perform manual labor. Such an energy balance framework is a potentially powerful tool for investigating weight control.

The conversion of white adipose tissue to brown or beige / bright adipose tissue has been reported as an effective mechanism for increasing energy consumption by utilizing excessive energy abundance. The role of brown adipose tissue (BAT) is well described in the following prior art:

Drugs and / or natural molecules that promote the conversion of white adipocytes to brown adipocytes are effective in the treatment / management of obesity-related conditions. However, we span various time scales to explain the components involved in energy expenditure, as well as the natural course of conditions such as obesity, and to estimate the scale and potential success of therapeutic interventions. Needs a better understanding of their interactions (Kevin D. Hall, Steven B. Heymsfield, Joseph W. Kemnitz, Samuel Klein, Dale A. Schoeller, and John R. Speakman, Energy balance and its components: implications for body weight regulation, Am J Clin Nutr. 2012 Apr; 95 (4): 989-994).
Recently, it has been observed that the gut microbiota is altered in conditions such as obesity and type II diabetes. Administration of probiotics to obese individuals resulted in effective weight loss. One particular gut microbiota, Akkermansia muciniphila, is inversely associated with obesity, diabetes, cardiac metabolic disorders, and mild inflammation (Cani et al., Next-Generation Beneficial Microbes:). The Case of Akkermansia muciniphila, Front. Microbiol., 22 September 2017, https://doi.org/10.3389/fmicb.2017.01765). A. It has been shown that the relative abundance of musiniphila increased by more than 100-fold after ingestion of prebiotics (Everard et al., 2014 Microbiome of prebiotic-treated mice reveals novel targets involved in host response during obesity). . ISME J. 8, 2116-2130. doi: 10.1038 / ismej2014.45). According to the research, A. It has also been shown that the number of muciniphila is lower in obese and type 2 diabetic mice and is increased with anti-diabetic treatment (Cani et al., Next-Generation Beneficial Microbes: The Case of Akkermansia muciniphila, Front. Microbiol., 22 September 2017, https://doi.org/10.3389/fmicb.2017.01765). According to another study, A. Musinifira treatment was observed to reverse hyperfat diet-induced metabolic disorders, including increased body fat mass, metabolic toxinemia, adipose tissue inflammation, and insulin resistance (Amandine Everard, Clara Belzer, Lucie Geurts). , Janneke P. Ouwerkerk, Celine Druart, Laure B. Bindels, Yves Guiot, Muriel Derrien, Giulio G. Muccioli, Nathalie M. Delzenne, Willem M. de Vos and Patrice D. Cani, Cross-talk between Akkermansia muciniphila and intestinal epithelium controls diet-induced obesity, PNAS May 28, 2013. 110 (22) 9066-9071). Therefore, increasing the viable count of Akkermansia muciniphila may be an effective treatment for the management of obesity, diabetes, and other metabolic disorders. The probiotic and beneficial effects of Akkermansia muciniphila are well documented in the following prior art literature.

Natural molecules are currently being widely evaluated for the management of obesity and related disorders. Extracts from Garcinia cambogia have been reported to have potential for weight loss. US Pat. No. 7063861 discloses a weight loss composition comprising garcinol and hydroxycitric acid (HCA), optionally with anthocyanins. US Pat. No. 8,329,743 also discloses a weight loss preparation containing garcinol, pterostilbene, and anthocyanins. US Pat. No. 7063861 shows that the combination of garcinol and HCA increases the bioavailability of HCA that provides anti-obesity effects. Thus, the anti-obesity effect of garcinol itself has not been reported and cannot be predicted from the prior art literature. In addition, Heo et al. (Gut microbiota Modulated by Probiotics and Garcinia cambogia Extract Correlate with Weight Gain and Adipocyte Sizes in High Fat-Fed Mice Sci Rep. Regulation of intestinal microflora with Cambodian extract and A. It reports an increase in Musinifira. The present invention solves the above problems by disclosing the anti-obesity effect of garcinol and the ability to regulate the intestinal microbiome.
A main object of the present invention is to disclose the anti-obesity effect of garcinol by providing weight loss and energy balance.
It is another object of the present invention to disclose the ability of garcinol to modify the intestinal microbiome to increase the viable count of the probiotic bacterium Akkermansia muciniphila.
Disclosing the lipid-lowering effect of garcinol is yet another object of the present invention.
The present invention serves such an object as described above and provides additional related advantages.

The present invention relates to a garcinol composition for obesity management. More specifically, the invention a presents the use of garcinol for a) maintaining energy balance in mammalian adipocyte systems, b) managing hypercholesterolemia, and c) reducing weight gain in mammals. Regarding. Also disclosed is the modification of the gut microbiota and the increase in the beneficial microorganism Akkermansia muciniphila by garcinol.

FIG. 5 is an oil-O-red staining of adipocytes showing a dose-dependent reduction in lipid accumulation in adipocytes with garcinol. It is a graph display of the inhibition rate of adipogenesis by galcinol. FIG. 6 is a graph showing a decrease in the expression of genes associated with adipogenesis in the garcinol-treated group. FIG. 6 is a graph showing increased expression of genes associated with brown adipose conversion and fat utilization in the garcinol-treated group. FIG. 6 is a graph showing changes in body weight of animals treated with different concentrations of garcinol over 4 months. FIG. 6 is a graph showing the final body weight of animals treated with different concentrations of garcinol for 120 days. It is a representative image showing different fat pad areas in the body of a mouse. FIG. 5 shows changes in the mass of adipose tissue around the peritoneum, mesentery, and perigonadal treated with different concentrations of garcinol. FIG. 6 is a graph showing the percentage reduction in visceral fat in a dose-dependent manner in animals treated with different concentrations of garcinol. FIG. 6 is a graph showing a decrease in the expression of genes associated with adipogenesis in the adipose tissue of animals treated with different concentrations of garcinol. FIG. 6 is a graph showing a decrease in the expression of genes associated with adipogenesis in the adipose tissue of animals treated with different concentrations of garcinol. FIG. 6 is a graph showing increased expression of genes associated with brown adipose tissue conversion and fat utilization in adipose tissue of animals treated with different concentrations of garcinol. FIG. 6 is a graph showing the serum total cholesterol and triglyceride levels of animals treated with different concentrations of garcinol. FIG. 6 is a graph showing the levels of LDL and VLDL in the sera of animals treated with different concentrations of garcinol. FIG. 6 is a graph showing the levels of HDL in the sera of animals treated with different concentrations of garcinol. It is a figure which shows the experimental design about the anti-obesity study with garcinol in the HFD (high-fat diet) -induced obesity mouse. It is a representative image showing the effect of garcinol on HFD-induced obesity in C57BL / 6 mice. Image A is a representative photograph of each group of mice at the end of the 13th week. Image B is a photograph of perigonadal adipose tissue, and image C is a photograph of the liver. FIG. 6 is a graphical representation of the body weight of animals treated with various concentrations of garcinol. Body weight is monitored weekly and the average body weight of each group is expressed as mean ± SE, p <0.05; a, b, c, and d are significantly different between each group. It is a photograph of adipose tissue around the gonads, retroperitoneum, and mesentery of animals treated with garcinol. It is a graph display of the adipose tissue mass of the animal to which garcinol was administered. It is a representative image of each study group about the pathological evaluation by H & E staining in the perigonadal adipose tissue. FIG. 6 is a graph showing the frequency percentage of adipocyte size in animals treated with garcinol. Adipocyte size was quantified under a microscope from a representative section. It is a figure which shows the change of the taxonomic composition of the colony bacterial community in the animal which received the garcinol. FIG. 16a shows changes in the relative abundance of the phylum of the fecal microflora, and FIG. 16b shows the relative abundance of the genus. It is a figure which shows the change of the taxonomic composition of the colony bacterial community in the animal which received the garcinol. FIG. 16a shows changes in the relative abundance of the phylum of the fecal microflora, and FIG. 16b shows the relative abundance of the genus. It is a figure which shows the principal coordinate analysis (PCoA) plot which shows the standardized relative abundance of the whole sample (A) genus (B). 6 is a heat map showing the abundance of 50 operational taxonomic units (OTUs) significantly altered by garcinol in HFD-fed mice. It is a figure which shows the effect of garcinol on the protein expression of adipocyte-specific factor and AMPK signaling in HFD feeding C57BL / 6 mouse peri-glandular adipose tissue. Protein levels of p-AMPK (Thr172), AMPK, Pref-1, SREBP-1, and PPARγ were determined by Western blot analysis with specific antibodies. β-actin was used as a load control. FIG. 6 is a graph representation of the level of adipocyte-specific factors and protein expression of AMPK signaling in HFD-fed C57BL / 6 mouse peri-glandular adipose tissue. Values indicate the relative density of protein bands standardized for β-actin. ^* p <0.05; ^** p <0.005; comparison with HFD treatment only. FIG. 6 is a graphical representation of the body weight of animals treated with various concentrations of rucinol and garcinol blends. It is a representative photograph of each group of mice at the end of the study period. FIG. 6 is a graph representation of perigonadal fat mass in animals treated with different concentrations of rucinol and a blend of rucinol. FIG. 6 is a graph representation of retroperitoneal fat mass in animals treated with different concentrations of rucinol and a blend of rucinol. FIG. 6 is a graph representation of mesenteric fat mass in animals treated with different concentrations of rucinol and a blend of rucinol.

In the most preferred embodiment, the present invention is a method for the treatment and management of obesity in a mammal, wherein an effective concentration of a garcinol-containing composition is administered to the mammal, and in the mammal, a) inhibition of adipogenesis. b) Disclose methods that include steps that result in weight and visceral fat loss. In a related embodiment, inhibition of adipogenesis is, but is not limited to, peroxisome proliferator-activated receptor gamma (PPARγ), CCAAT / enhancer binding protein alpha (cEBPα), first apoptotic signal (FAS). ), Adipocyte protein 2 (AP2), resistin, and leptin are brought about by downregulation of genes selected from the group. In a related embodiment, inhibition of lipogenesis is, but is not limited to, phosphoadenosine monophosphate-activated protein kinase (p-AMPK), AMP-activated protein kinase (AMPK), and pre-lipogenic factor 1 (PREF-). It is brought about by the upward regulation of genes selected from the group consisting of 1). In another related embodiment, the visceral fat is selected from the group consisting of mesenteric fat, peritoneal fat, and peri-gonadal fat. In a related embodiment, the composition is formulated with a pharmaceutically / nutritionally acceptable excipient, adjunct, diluent, or carrier, tablets, capsules, syrups, gummy, powders, suspensions. Orally administered in the form of emulsions, syrups, candy, and groceries.

In another preferred embodiment, the invention is a method of achieving energy balance in a mammalian adipocyte system, wherein an effective amount of a garcinol-containing composition targeted to premammalian adipocytes and / or adipocytes is administered. , (A) the effect of increasing inhibition of fat production, and (b) specifically recruiting brown adipocytes or brown-like (beige or bright) adipocytes, (c) brown-like phenotype in white adipocyte storage sites. Methods comprising achieving the effect of increasing expression of factors that function individually or in combination to induce (beige or bright adipocytes), resulting in fat utilization and energy balance effects in said mammal. Disclose. In a related embodiment, the factor comprises a transmembrane protein mitochondrial uncoupling protein (UCP-1), a transcription co-regulator that controls genes involved in energy metabolism, protein 16. PR domain (PR domain proteining protein 16) (PRDM16) and peroxysome growth factor activated receptor gamma coactivator 1-alpha (Peroxysome proteinor-activated receptor gamma coactivator 1-alpha) (PG) (PG) Bone-forming protein 7 (BMP7), which is a secreted protein, is mentioned. In a related embodiment, the composition is formulated with a pharmaceutically / nutritionally acceptable excipient, adjunct, diluent, or carrier, tablets, capsules, syrups, gummy, powders, suspensions. Orally administered in the form of emulsions, syrups, candy, and groceries.

In another preferred embodiment, the invention is a method of modifying the intestinal microbial flora in a mammal, wherein an effective amount of a garcinol-containing composition is administered to the mammal to bring about a change in the intestinal microbial flora. Disclose methods, including. In a related embodiment, the gut microbiota is the phylum Deferribacters, the phylum Pseudomonadota, the phylum Bacteroidota, the phylum Verrucomicrobiota, and the phylum Firmicutes (Firmicutes). Will be done. In another related embodiment, the intestinal microbial flora is Lactobacillus, Bacteroides, Clostridium, Anaerobranca, Disgonomonas, Dysgononas, and Dysgononas. ), Ruminococcus, Bacteroides, Oscillosspira, Parabacterrodies, Ackermanisa, and Blautia. More specifically, the intestinal microbial flora includes Parabacteroides goldsteini, Bacteroides caccace, Johnsonella Ignavas (Johnsonella ignavela), Brunelava ignavela, and Brautia lignawe. （Ｄｙｓｇｏｎｏｍｏｎａｓ ｗｉｍｐｅｎｎｙｉ）、ブラウティア・ハンセンニ（Ｂｌａｕｔｉａ ｈａｎｓｅｎｎｉ）、アナエロブランカ・ザバルチンニ（Ａｎａｅｒｏｂｒａｎｃａ ｚａｖａｒｚｉｎｎｉ）、オシロスピラ・エアエ（Ｏｓｃｉｌｌｏｓｐｉｒａ ｅａｅ）、ムシスピリルム・シェドレリー（Ｍｕｃｉｓｐｉｒｉｌｌｕｓ ｓｃｈａｅｄｌｅｒｉ）、ブラウティア・コッコイデス（Ｂｌａｕｔｉａ ｃｏｃｃｏｉｄｅｓ）、アナエロツルンカス・コリホミニス（Ａｎａｅｒｏｔｒｕｎｃｕｓ ｃｏｌｉｈｏｍｉｎｉｓ）、ブチリビブリオ・プロテオクラスティカス（Ｂｕｔｙｒｉｖｉｂｒｏ ｐｒｏｔｅｏｃｌａｓｔｉｃｕｓ）、アッカーマンシア・ムシニフィラ（Ａｋｋｅｒｍａｎｓｉａ ｍｕｃｉｎｉｐｈｉｌａ）、ラクノスピラ・ペクチノスチザ（Ｌａｃｈｎｏｓｐｏｒａ ｐｅｃｔｉｎｏｓｃｈｉｚａ）、ペドバクター・クワンジャンゲンシス（Ｐｅｄｏｂａｃｔｅｒ ｋｗａｎｇｙａｎｇｅｎｓｉｓ）、アルカリフィルス・クロトンアトキシダンス（Ａｌｋａｌｉｐｈｉｌｕｓ ｃｒｏｔｏｎａｔｏｘｉｄａｎｓ）、ラクトバチルス・サリバリウス（ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｌｉｖａｒｉｕｓ）、アナエロビブリオ・リポリティカス（Ａｎａｅｒｉｖｉｂｒｉａ ｌｉｐｏｌｙｔｉｃｕｓ）、ロドサーマス・クラルス（Ｒｈｏｄｏｔｈｅｒｍｕｓ ｃｌａｒｕｓ）、バクテロイデス・ステルコリロソリス（Ｂａｃｔｅｒｏｉｄｅｓ ｓｔｅｒｃｏｒｉｒｏｓｏｒｉｓ）、ルミノコッカス・フラベファシエンス(Ruminococcus flavefaciens), Bacteroides xylanisolvens, Ruminococcus gnavus, Clostridium termitidis. ｄｉｓ）、クロストリジウム・アルカリセルロシー（Ｃｌｏｓｔｒｉｄｉｕｍ ａｌｋａｌｉｃｅｌｌｕｌｏｓｉ）、エムチシシア・オリゴラフィカ（Ｅｍｔｉｃｉｃｉａ ｏｌｉｇｏｒａｐｈｉｃａ）、シュードブチリビブリオ・キシラニボランス（Ｐｓｅｕｄｏｂｕｔｙｒｉｖｉｂｒｏ ｘｙｌａｎｉｖｏｒａｎｓ）、アクチノマイセス・ナツラエ（Ａｃｔｉｎｏｍｙｃｅｓ ｎａｔｕｒａｅ）、ペプトニフィラス・コキシー（Ｐｅｐｔｏｎｉｐｈｉｌｕｓ ｃｏｘｉｉ）、 And Dorichospermum curvum. In a related embodiment, alteration of the intestinal microbial flora is a disease selected from the group consisting of obesity, cardiovascular complications, inflammatory bowel disease, Crohn's disease, celiac disease, metabolic syndrome, liver disease, and neuropathy. It is effective in the treatment management of. In a related embodiment, the composition is formulated with a pharmaceutically / nutritionally acceptable excipient, adjunct, diluent, or carrier, tablets, capsules, syrups, gummy, powders, suspensions. Orally administered in the form of emulsions, syrups, candy, and groceries.

In another preferred embodiment, the invention is a method for increasing the viable count of Akkermansia muciniphila in the intestine of a mammal by administering to the mammal an effective amount of a garcinol-containing composition. Disclose a method, including steps that result in an increase in colonies. In a related embodiment, an increase in the number of colonies of Akkermansia muciniphila causes weight loss through the AMPK signaling pathway by causing endocannabinoid release. In a related embodiment, the composition is formulated with a pharmaceutically / nutritionally acceptable excipient, adjunct, diluent, or carrier, tablets, capsules, syrups, gummy, powders, suspensions. Orally administered in the form of emulsions, syrups, candy, and groceries.

In another preferred embodiment, the present invention is a method of treating and managing hyperlipidemia in mammals, wherein an effective concentration of garcinol-containing composition is administered in the blood of said mammal (i) total blood. Decreasing the amount of cholesterol levels; (ii) decreasing the concentration of low-density lipoprotein (LDL) and very low-density lipoprotein (VLDL); (iii) increasing the concentration of high-density lipoprotein (HDL) That; as well as (iv) disclose methods comprising the steps that have the effect of lowering the concentration of serum triglycerides. In a related embodiment, the medical cause of hyperlipidemia is obesity. In a related embodiment, the composition is formulated with a pharmaceutically / nutritionally acceptable excipient, adjunct, diluent, or carrier, tablets, capsules, syrups, gummy, powders, suspensions. Orally administered in the form of emulsions, syrups, candy, and groceries.
In another preferred embodiment, the invention discloses a garcinol-containing composition for use as a prebiotic agent.
Hereinafter, specific exemplary examples are included that articulate the most preferred embodiments.

(Example 1)
Anti-obesity effect of garcinol-Sami Labs Limited, Bangalore, India and Srimad Andavan Arts & Science College, Tiruchyrapalli, India research on fat-producing solutions in culture 3T3L1 and animal tissue. Preparation of 10 mg / ml garcinol (20%) stock was prepared in DMSO and filtered through a 0.2 μm syringe filter. The stock was diluted 1000-fold in DMEM to obtain a final concentration of 10 μg / ml and serially diluted. Insulin (Hi Media) was used as a solution at a concentration of 20 mg / ml. This was diluted to 1 μg / ml in DMEM. A stock of IBMX (Sigma) -5 mM was prepared in DMEM and diluted 10-fold for use at a final concentration of 0.5 mM. A stock of dexamethasone (Sigma) -10 μM was prepared in DMEM and diluted 40-fold to obtain a final concentration of 0.25 μM.

Cell culture Mouse 3T3-L1 preadipocytes were cultured at 37 ° C. and 5% CO2 in DMEM containing 25 mM glucose with 10% heat-inactivated fetal bovine serum and antibiotics. When the cells were 70-80% confluent, they were trypsinized, washed and seeded in 6-well plates at a density of 2 × 10 ⁶ cells per well. Two days after reaching confluence (day 0), 1 μg / mL insulin, 0.25 μM dexamethasone, 0.5 mM 1-methyl-3-isobutyl-xanthine (1 μg / mL insulin, 0.25 μM dexamethasone, 0.5 mM 1-methyl-3-isobutyl-xanthine) in DMEM medium containing 10% fetal bovine serum (FBS). IBMX) and different concentrations of garcinol (20%) were added to induce cells to differentiate. From day 3 to day 7, cells were maintained in advanced medium supplemented with 1 μg / mL insulin and different concentrations of garcinol (20%). Untreated cells and undifferentiated cells grown in FCS medium were used as adipogenic positive and negative controls for this experiment. The amount of triglyceride accumulated in adipocytes was quantified by Oil Red O staining.

RNA Extraction After the second progression on day 7, cells were harvested and total RNA was extracted using the Trizol method. The extracted RNA was treated with Deoxyribonuclease I to remove any contaminating DNA and again extracted using phenol: chloroform: isoamyl alcohol extraction (24:25: 1). RNA quality was determined by checking the absorbance at 260/280 nm using Nanodrop (Thermo).
Gene Expression Studies in Mouse Fat Pads Frozen fat pads from treated and untreated animals were collected and frozen for later RNA. Approximately 100 mg of tissue was homogenized in ice and extracted with 1 ml Trizol as previously described.

Quantitative real-time PCR
2 μg of total RNA was obtained using SuperScript III First-Strand Synthesis System (Life Technologies) for cDNA synthesis. Quantitative RT-PCR analysis was performed on Roche Light cycler 96 using the SYBR Green Master Mix (Thermo Scientific) to determine the expression of brown adipose-specific genes. β-actin was used as a housekeeping gene. The relative RNA abundance of the BAT gene was standardized for the housekeeping β-actin gene and expressed as ΔΔCT (equivalent to a multiple change converted by Log ₂ ).

Primer sequence: Primers used to determine the expression of brown adipose-specific genes associated with adipogenesis are shown in Table 1.

Results Lipids accumulated in adipocytes were quantified by Oil Red O staining. Garcinol showed a dose-dependent reduction in lipid accumulation in adipocytes (Fig. 1), with 5 μg / ml and 10 μg / ml showing the highest inhibition of lipid accumulation at 47.8% and 47.2% (Fig. 1). 1b).

With respect to genes involved in adipogenesis, PPARγ is considered to be a major regulator of adipogenesis. Decreased expression of PPARγ reduces the expression of other adipogenesis-specific genes. In this study, garcinol showed a dose-dependent reduction in PPARγ expression and expression of genes associated with adipogenesis and fatty acid synthesis such as cEBPα, FAS, and AP2 (Fig. 2), with garcinol in a dose-dependent manner. It has been shown to inhibit adipogenesis.

Garcinol also significantly increased brown adipose tissue-specific genes. Expression of UCP1, PRDM16, PGC1α, and BMP7 was increased in a dose-dependent manner by garcinol (FIG. 3), where garcinol converted the white adipose tissue storage site to brown or bright / beige adipose tissue, thereby. It suggests that it is effective in increasing energy expenditure due to fat utilization and lipolysis.

Effect of Garcinol on High-Fat Diet-Induced Obesity in C57 Mice Methods Animals-6-8 week old C57 / BL6 mice were used in this study in 8 animals / group (4 males and 4 females). Animals are air-conditioned with adequate fresh air supply (12-15 air exchanges per hour), room temperature 20.2-23.5 ° C, and relative humidity 58-64%, 12 hours of fluorescent light. And were bred under standard experimental conditions in a 12 hour dark cycle. Temperature and relative humidity were recorded once daily.
Prey animals were fed a normal diet (9 kcal / day) and a high-fat diet (50 kcal / day) throughout the acclimation and experimental period.
Water was fed to the animals with a high-fat diet throughout the acclimatization and experimental periods. Water from a water strainer with a purifier was provided in an animal feeding bottle with a stainless steel shipper tube.

All studies were performed after obtaining the required approval from the Commission (approval number: 790/03 / ac / CPCSEA) in accordance with the CPCSEA ethical guidelines.
a. Committee for the Management and Monitoring of Experiments on Animals for Laboratory Animal Facilities published in the December 15, 1998 Gazette of India (CPCSE) (CPCSE) (CPCSE) According to the recommendation of.
b. The CPCSEA approval number for this study (anti-obesity activity) is SAC / IAEC / BC / 2017 / IP. -001.

The design of the study group is shown in Table 2.

Animal body weights were recorded on all days of the experimental period. At the end of the experimental period, animals were sacrificed by cervical dislocation. Blood was collected and serum was separated by centrifugation and used for analysis of biochemical parameters. Organs such as the liver, kidneys, spleen, and pancreas and adipose tissue (retroperitoneal fat, perigonal fat, and mesenteric fat) were resected and washed in phosphate buffered saline.

Efficacy measurement The following parameters were measured in the above group.
Measurement of body weight Determination of organ mass Cholesterol estimation (Zak et al., (2009) A new method for the determination of serum cholesterol. J Clin Endocrinol Metab., 94 (7), 2215-2220)
Foster LB and Dunn RT (1973) Stable reagents for determination of serum triglycerides by a colorimetric Hantzsch condensation method. Clin Chem, 196, 338-340)
Estimating HDL Cholesterol (Burstein et al., (1970). Determination of HDL cholesterol. J.lipid Res., 11, 583)
Determination of LDL and VLDL (Friedewald et al., (1972) Estimation of the concentration of Low Density Lipoprotein cholesterol in plasma without use of preparative centrifuge. J. Clin Chem .; 18: 499)

Results Weight Results showed that garcinol inhibits weight gain in a dose-dependent manner in animals fed a high-fat diet for 120 days (FIGS. 4a and 4b). The percentage changes in body weight are shown in the table below.

Decreased fat storage sites Fat depletion in different fat pad areas of mice (Fig. 5) was also evaluated. The masses of the retroperitoneal, perigonadal, and mesenteric fat storage sites after 120 days of administration of garcinol are tabulated as follows.

ガルシノール処置は、異なる脂肪パッド領域における脂肪蓄積を有意に低下させた（図６）。内臓脂肪のパーセンテージは、ガルシノール処置により低下し（図７）、１０ｍｇ／ｋｇ体重の用量が最大効果を示した。

Garcinol treatment significantly reduced fat accumulation in different fat pad areas (Fig. 6). The percentage of visceral fat was reduced by garcinol treatment (Fig. 7), with a dose of 10 mg / kg body weight showing maximum effect.

Organ Mass Garcinol administration does not adversely affect organ mass, suggesting that Garcinol does not induce any adverse effects in critical organs (Table 5).

Gene expression: Decreased expression of genes associated with lipogenesis was observed in the fat pads of animals treated with garcinol. Similar to the mouse 3T3-L1 cell line, garcinol administration significantly reduced the expression of PPARγ, AP2, FAS, resistin, and leptin in the fat pad region (FIGS. 8a and 8b). Similarly, garcinol administration effectively increased the expression of brown adipose-specific genes in the mouse fat pad region (Fig. 9).
Lipid profile: A high-fat diet increased the levels of total cholesterol, LDL, VLDL, and triglycerides in the serum of study animals. A high-fat diet co-administered with garcinol significantly reduced serum total cholesterol and triglycerides (FIG. 10a), LDL and VLDL (FIG. 10b) and increased HDL levels (FIG. 10c).

Conclusion:
Garcinol treatment showed a dose-dependent inhibition of adipogenesis in vitro and induced a conversion of white adipose tissue to brown or bright / beige adipose tissue, thereby increasing fat utilization and energy metabolism. In vivo results show that garcinol is effective in significantly reducing body weight and visceral fat accumulation at 10 mg / kg, and reduces lipogenesis-specific gene expression in the fat pad in mouse fat pads, resulting in brown adipose tissue. It has been shown to increase specific genes. Garcinol administration also results in a decrease in visceral fat and organ mass, indicating that galcinol promotes lipolysis and energy metabolism. Overall, garcinol induces weight loss, lowers visceral fat, and maintains the health of vital organs.

(Example 2)
Anti-obesity effect of garcinol-Research methodologies performed in National Taiwan University, Taipei, Taiwan Reagents and antibodies AMPK and p-AMPK (Thr172) antibodies were purchased from Cell Signaling Technology, Beverly, MA. The SREBP-1 antibody was procured from Santa Cruz Biotechnology (Santa Cruz, CA, USA). PPARγ and Pref-1 antibodies were purchased from abcam (Cambridge, England). Mouse β-actin monoclonal antibody was obtained from Sigma Chemical Co (St. Louis, MO, USA). Bio-Rad protein assay dye reagents were purchased from Bio-Rad Laboratories (Munich, Germany). Xylene and hematoxylin and eosin (H & E) stains were obtained from Surgipath (Peterborough, UK). Cholesterol used as part of the animal diet was obtained from Acros Organics (Bridgewater, NJ, USA). Garcinol was added to Sabinsa Corp. Procured from (East Windsor, NJ, USA). The purity of garcinol was determined by high performance liquid chromatography (HPLC) to be greater than 99%.

Animal Breeding and Experimental Design Five-week-old male C57BL / 6 mice were purchased from BioLASCO Experimental Animals Center (Taiwan Co., Ltd, Taiwan, Taiwan, Taiwan) and under controlled atmosphere (25 ± 1 ° C, 50% relative humidity). , 12 hours light / dark cycle. After 1 week of acclimation, the animals were subjected to the normal diet (ND, 15% energy as fat) group, HFD (50% energy as fat) group, and 0. They were randomly assigned to the HFD group with 1% or 0.5% garcinol. The experimental design is summarized in Figure 11. The experimental diet was modified from the Purina 5001 diet (LabDiet, PMI Nutrition International, St. Louis, MO, USA). Animals had free access to food and water. The food cup was refilled with fresh food daily. All animal experiment protocols used in this study were approved by the Instituteal Animal Care and Use Committee of the National Taiwan University (IACUC, NTU). At the end of the study, animals were slaughtered and dissected by CO ₂ choking and their whole body mass, as well as liver, kidney, spleen, adipose tissue (peri-gonadal fat, retroperitoneal fat, and mesenteric fat), and Selected tissues containing serum were immediately collected, weighed and photographed.

Histopathological test Peri-glandular fat and a portion of the middle lobe of the liver were excised, fixed in 10% buffered formalin, dehydrated with a series of ethanol solutions and treated for paraffin embedding. It was cut into 3-5 μm thick sections, deparaffinized, rehydrated, stained with H & E and subjected to micrograph evaluation. Adipocyte size was determined using ImageJ software (ImageJ, WS, ImageJ, US National Institutes of Health, Bethesda, MD, USA).

Biochemical analysis Blood samples were collected from the left ventricle under anesthesia. The sample was mixed in 10 μL of sodium heparin and centrifuged at 3500 rpm for 10 minutes at 4 ° C. Plasma was then stored at −80 ° C. until use. Levels of glutamate pyruvate transaminase (GPT), total cholesterol (TC), TG, high density lipoprotein (HDL), and low density lipoprotein (LDL) at National Laboratory Animal Center, NLAC (Taipei, Taiwan) 7080 Biochem (Hitachi, Tokyo, Japan) analyzed according to the manufacturer's instructions.

16S rDNA gene sequencing and analysis Total DNA was extracted from fresh fecal samples. Purified DNA was eluted using the innuSPEED Store DNA kit (Anallytic Jena AG, Jena, Germany) according to the manufacturer's protocol. Caporaso et al. (Caporaso, JG, Lauber, CL, Walters, WA, Berg-Lyons, D. et al., Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample. Proc.Natl.Acad.Sci US A Using the PCR primer sequence from 2011, 108 Suppl 1, 4516-4522), the 16S rRNA variable region was amplified and the PCR conditions were set to Tung et al. (Tung, YC, Lin, YH, Chen, HJ, Chou, SC et). al., Piceatannol Exerts Anti-Obesity Effects in C57BL / 6 Mice through Modulating Adipogenic Proteins and Gut Microbiota. Molecules. 2016, 21) and Chou et al. (Chou, YC, Suh, JH, Wang, Y., Pahwa, M. et. Al., Boswellia serrata resin extract alleviates azoxymethane (AOM) / dextran sodium sulfate (DSS)-induced colon tumorigenesis. Mol. Nutr Food Res. 2017, 61). Then, using the unit duplication sequence, an index labeling library was constructed with the Illumina DNA Library Preparation Kit (Illumina, San Diego, CA, USA). Illumina MiniSeq NGS System (Illumina) was used to analyze more than 100,000 reads in paired-end sequencing (2 x 150 bp), and the metagenomics workflow was a database of 16S rRNA data. Organisms were classified from the unit replication sequence using. The classification was based on the Greengenes database (https://greengenes.lbl.gov/). The output of the workflow was the classification of readings at several taxonomic levels: kingdom, phylum, class, order, family, genus, and species.

Protein preparation and Western blot tissue, ice-cold lysis buffer (10% glycerol, 1% TritonX-100, 1 mM Na ₃ VO ₄ , 1 mM EGTA) containing one Protease Inhibitor Cocktail Table (Roche, Indianapolis, IN, USA). Homogenize on ice for 1 hour in 10 mM NaF, 1 mM Na ₄ P ₂ O ₇ , 20 mM Tris buffer (pH 7.9), 100 μM β-glycerophosphate, 137 mM NaCl, and 5 mM EDTA), then at 17,500 g. It was centrifuged at 4 ° C. for 30 minutes. The protein concentration was measured by the Bio-Rad protein assay (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Statistical analysis A statistical assessment of the significance of differences between groups of mice was assessed using Student's t-test. For experiments comparing multiple groups, the differences were analyzed by performing one-way analysis of variance (ANOVA) and Duncan's post-test. Data were presented as the mean ± SE for the indicated number of independently performed experiments, with a p-value <0.05 considered to be statistically significant. Principal component analysis (PCA) was performed to visualize the differences between the samples.

Results Weight Gain Results showed that 13 weeks of HFD feeding resulted in a significant increase in body weight and liver mass along with perigonal, retroperitoneal, and mesenteric fat accumulation. Diets supplemented with 0.1% and 0.5% garcinol reduced body weight in a dose-dependent manner. Simultaneous treatment with a high dose of garcinol (0.5%) and an HFD diet inhibited weight gain and there was no difference between the HFD + 0.5% garcinol group and the ND group (FIGS. 12 and 13).

Effect on White Adipose Tissue Adipocyte Size and Liver Homeostasis Garcinol at a concentration of 0.5% compared the fat mass of all three white adipose tissues with the HFD group, 85.1 with respect to the mass around the gonads. %, 92.4% for retroperitoneal mass, and 77.7% for mesenteric mass, dramatically reduced (FIGS. 14a and 14b).

The average adipocyte size in perigoneral adipose tissue was evaluated by H & E staining, and the results revealed that adipocytes were enlarged in HFD-fed mice compared to adipocytes in ND mice. rice field. The increase in adipocyte size was significantly reduced in garcinol-treated mice (Fig. 15). Garcinol (0.5%) was able to prevent HFD-induced adipocyte expansion, and adipocytes were distributed in a size of 2000 μm ² . Importantly, adipocyte size can be prevented or inhibited in a dose-dependent manner by garcinol (Table 6).

４つの群の間での差の有意性は、一元配置ＡＮＯＶＡおよびＤｕｎｃａｎの多範囲検定により分析された。異なる文字を有する値は、各群間で有意に異なっている（ｐ＜０．０５）。

The significance of the differences between the four groups was analyzed by one-way ANOVA and Duncan multi-range tests. Values with different letters are significantly different between the groups (p <0.05).

データは、平均±ＳＥとして提示されている。４つの群の間での差の有意性は、一元配置ＡＮＯＶＡおよびＤｕｎｃａｎの多範囲検定により分析された。同じ行において同じ上付き文字を共有しない値は、群の間で有意に異なっている。ｐ＜０．０５；ａ、ｂ、ｃ、およびｄは、各群間で有意に異なっている。 Lipid Profiles: Plasma lipid profiles have also been analyzed and presented in Table 7.

The data are presented as mean ± SE. The significance of the differences between the four groups was analyzed by one-way ANOVA and Duncan multi-range tests. Values that do not share the same superscript on the same line are significantly different between groups. p <0.05; a, b, c, and d are significantly different between the groups.

Mice treated with garcinol at 0.1% and 0.5% significantly reduced serum levels of both TC and TG. With respect to LDL and HDL, garcinol (0.1% and 0.5%) was able to reduce HFD-induced LDL levels in a dose-dependent manner. Since the TC reduction was brought about by HFD, the HFD group increased not only LDL levels, but also HDL levels. Therefore, we used the LDL / HDL ratio to represent this change. High and low doses of garcinol can significantly reduce the LDL / HDL ratio as compared to the HFD group.

Garcinol assessed the overall composition of the bacterial community in different groups that reversed HFD-induced dysbiosis by analyzing the degree of taxonomic similarity between the metagenus samples at the genus level. Bacterial communities were clustered using PCA, which identified microbial communities based on HFD diet / garcinol treatment. The intestinal microflora of obese humans and HFD-fed mice is characterized by an increase in Firmicutes (F / B ratio) relative to Bacteroides (Brun, P., Castagliuolo, I., Di, L., V, Buda, A. et al., Increased intestinal permeability in obese mice: new evidence in the pathogenesis of nonalcoholic steatohepatitis. Am.J Physiol Gastrointest.Liver Physiol 2007, 292, G518-G525). The results showed that the phylum level of the HFD group had a higher F / B ratio compared to the ND group (FIG. 16a). Interestingly, Garcinol treatment reduced the F / B ratio by significantly increasing the Bacteroides phylum community. In addition, galcinol treatment increased the number of Belcomicrobium phylum communities (Fig. 16b). A pairwise comparative PCA based on the UniFrac of the community structure disclosed the distribution of the microbial community among the four mouse groups. The main finding of PCA was that different diets promoted the development of various gut microbiota communities. HFD-fed mice formed different clusters than ND-fed mice, and HFD-fed mice were also different from galcinol-treated mice (FIGS. 17a, b, and c). However, microbial communities in mice treated with high doses of garcinol (0.5%) formed clusters in close proximity to those in ND mice, which caused garcinol to have a significant effect on intestinal microbial community composition. It also shows that it reversed HFD-induced dysbiosis.

Effect of Garcinol Administration on Intestinal Microbial Community Composition To further investigate whether changes in the intestinal microbial flora were induced by garcinol supplementation, we then determined the genus level of the intestinal microbial flora. A heat map was used to represent the abundance of 50 OTUs significantly altered by garcinol in HFD-fed mice (FIG. 18). The results showed that HFD-fed mice increased the Brautia community, which was dramatically reduced in both the high-dose and low-dose garcinol-treated groups. By this study, Blautier spp. And Enterobacter spp. However, it was pointed out that it is associated with obesity caused by HFD in a mouse model (Becker, N., Kunath, J., Loh, G., and Blaut, M. Human intestinal microbiota: characterization of a simplified and stable. gnotobiotic rat model. Gut Microbes. 2011, 2, 25-33; Fei, N. and Zhao, L. An opportunistic pathogen isolated from the gut of an obese human causes obesity in germfree mice. ISME.J 2013, 7, 880- 884). Interestingly, the genera Bacteroides, Bacteroides, and Akkermansia also increased dramatically in Garcinol-fed mice. The genera Bacteroides and Bacteroides belong to the phylum Bacteroides, and the genus Akkermansia belongs to the phylum Belcomicrobium: this is why the F / B ratio moved as it did after induction with Garcinol treatment. It explains that. In the heat map, we found that the assemblages of Anaeroblanca zabaltinni, Blautia coccoides, and Butilivibrio proteoclasticus had increased post-HFD numbers, but garcinol administration was not only for those bacteria. It also reduced Osirospira aire, Musispyrilum shedrery, Anaeroturunkas colihominis, and Lachnospiraceae pectinostiza. In addition, Garcinol increased the numbers of Akkermansia muciniphila, Bacteroides stellolilosolis, and Bacteroides xylanisolvens, which decreased in the ND and HFD groups.

Anaeroblanca zabaltinni, Blautier coccoides, and Butilivibrio proteoclasticus belong to the Firmicutes phylum; Anaeroblanca zabaltinni positively correlates with IBD patients and Blautier coccoides is increased in the HFD-induced mouse model. Butilivibrio proteoclasticus is extremely sensitive to the toxic effects of unsaturated fatty acids and has been associated with obesity. On the other hand, Bacteroides stellolilosolis and Bacteroides xylanisolvens belong to the Bacteroides phylum, and Akkermansia muciniphila belongs to the Belcomicrobium phylum. Andoh et al. (Andoh, A., Nishida, A., Takahashi, K., Inatomi, O. et al., Comparison of the gut microbial community between obese and lean peoples using 16S gene sequencing in a Japanese population. J Clin. Biochem .Nutr 2016, 59, 65-70) performed a 16S rRNA sequence analysis of the intestinal microflora profile of obese and lean Japanese populations, who found that Bacteroides stellolilosolis was lean Japanese. Found to exist in. Liu et al. (Liu, R., Hong, J., Xu, X., Feng, Q. et al., Gut microbiome and serum metabolome alterations in obesity and after weight-loss intervention. Nat.Med 2017, 23, 859- 868) conducted metabolome-wide related studies and serum metabolomics profiling in young Chinese individuals of lean and obese. They linked intestinal microbial flora changes to circulating amino acids and obesity, indicating that Bacteroides xylanisolvens is significantly abundant in lean controls.

Several studies have highlighted the effects of the mucin-degrading bacterium Akkermansia muciniphila, which is more abundant in the mucosa of healthy subjects, in the mucosa of diabetic patients or animals (Liou, AP., Paziuk). , M., Luevano, J.M., Jr., Machineni, S. et al., Conserved shifts in the gut microbiota due to gastric bypass reduce host weight and adiposity. Sci Transl.Med 2013, 5, 178ra41). Many studies have demonstrated the dietary effects of Akkermansia muciniphila and how it also inhibits obesity. Dietary supplementation of HFD with grape polyphenols resulted in a decrease in F / B ratio and dramatic changes in the structure of the gut microbiota, including the bloom of Akkermansia muciniphila (Roopchand, D.E., Carmody, R.N., Kuhn, P., Moskal, K. et al., Dietary Polyphenols Promote Growth of the Gut Bacterium Akkermansia muciniphila and Attenuate High-Fat Diet-Induced Metabolic Syndrome. Diabetes 2015, 64, 2847-2858). All these studies support the suggestion that Akkermansia muciniphila has a potential role as a probiotic with anti-obesity effects, and therefore we show that garcinol has a prebiotic role. Advocate that.

Garcinol treatment was performed by Akkermansia spp. We affected the AMPK signaling pathway by increasing the number of and inducing endocannabinoid expression. We further investigated the molecular mechanism by which garcinol exerts an anti-obesity effect. Protein levels of AMPK, p-AMPK, PPARγ, preadipocyte factor 1 (Pref-1), and SREBP-1 in HFD-fed C57BL / 6 mice are shown in FIG. HFD feeding produced a decrease in AMPK in white adipose tissue compared to AMPK in the ND group, which was increased by administration of low doses of garcinol (0.1%). Interestingly, high doses of garcinol (0.5%) did not increase AMPK protein levels or p-AMPK protein levels. We have found that this is Akkermansia spp. It was estimated that it may be related to. 4 weeks of A. HFD-induced obese mice. Administration of Musinifera improved the content of endocannabinoids, including 2-AG, 2-PG, and 2-OG (Everard, A., Belzer, C., Geurts, L., Ouwerkerk, JP et al., Cross-talk between Akkermansia muciniphila and intestinal epithelium controls diet-induced obesity. Proc.Natl.Acad.Sci US A 2013, 110, 9066-9071). Within the intestinal tissue, an increase in 2-AG reduces metabolic toxinemia and systemic inflammation by increasing goblet cells and Treg populations. However, in perigoneral adipose tissue, an increase in 2-AG also stimulates preadipocyte differentiation (via upregulation of adipocyte PPARγ levels) and above stimulation of lipoprotein lipase and FAS levels and glucose uptake. Storage of adipose tissue by enhancing novel fatty acid synthesis (through regulation), reducing fatty acid oxidation (through inhibition of AMPK), and enhancing triacylglycerol accumulation (through inhibition of adipose degradation). Increased capacity. 2-AG is a phospholipid-derived lipid containing an arachidonic acid chain within its chemical structure. 2-AG is also an intermediate in triacylglycerol and phospholipid metabolism, and therefore mice treated with HFD can readily supply substrates for 2-AG production. Pref-1 has been identified as an inhibitor of adipocyte differentiation that is highly expressed in preadipocytes and disappears during differentiation. Garcinol treatment causes an increase in protein levels of Pref-1 in epididymal adipose tissue, suggesting that garcinol may function to maintain preadipocyte status in HFD-fed mice.

Conclusion The results show that Garcinol treatment results in unexpected changes in the composition of the intestinal microbial flora in mice undergoing HFD, which changes can affect the underlying molecular mechanism. In addition, these findings reinforce the notion that changes in the gut microbial community aimed at increasing the Akkermansia population can prevent HFD-induced obesity.

(Example 3)
Comparative Evaluation of Weight Loss of Garcinol and Compositions Containing Garcinol, Pterostilbene, and Anthocyanins The present invention compares to compositions containing garcinol, pterostilbene, and anthocyanins (Garcinol Blend (GB)) in mammals. We studied the anti-obesity effect of stilbene. The study was performed in vivo in 5 week old C57BL / 6 male mice. A total of 42 mice were included in this study in 6 groups of 7 mice each. The groups were divided as shown in Table 8.

体重は毎週、モニターされ、各群（ｎ＝７）の平均体重は、平均±ＳＥとして表された。６つの群の間での差の有意性は、一元配置ＡＮＯＶＡおよびＤｕｎｃａｎの多範囲検定により分析された。ｐ＜０．０５、ａ、ｂ、およびｃは、各群間で有意に異なっている。 The high-fat diet (HFD) group was given a 45% high-fat diet for 16 weeks to induce obesity, while at the same time receiving the test substance as shown in the table above. The normal group was given a normal diet for 16 weeks.

Body weight was monitored weekly and the average body weight for each group (n = 7) was expressed as mean ± SE. The significance of the differences between the 6 groups was analyzed by one-way ANOVA and Duncan multi-range tests. p <0.05, a, b, and c are significantly different between the groups.

The results showed that the mouse group fed with HFD + 0.5% garcinol showed the most significantly reduced body weight and prevented weight gain compared to the HFD feeding group and the HFD + GB group (FIG. 19a and FIG. 19a and). 19b). Mice treated with HFD + 0.5% garcinol showed the least weight gain compared to the other groups (Table 9), which is an unexpected finding and cannot be predicted by one of ordinary skill in the art.

各群（ｎ＝７）の平均体重は、平均±ＳＥとして表されている。６つの群の間での差の有意性は、一元配置ＡＮＯＶＡおよびＤｕｎｃａｎの多範囲検定により分析された。同じ行において同じ上付き文字を共有しない値は、群の間で有意に異なっている。ｐ＜０．０５、ａ、ｂ、およびｃは、各群間で有意に異なっている。 The effects of rucinol and garcinol blends on reducing the mass of adipose tissue around the peritoneum, retroperitoneum, and mesentery were also evaluated. The results showed that 0.5% rucinol significantly reduced the mass of adipose tissue around the gonads, retroperitoneum, and mesentery compared to the garcinol blend (FIGS. 20a, b, c). ).

The average body weight of each group (n = 7) is expressed as an average ± SE. The significance of the differences between the 6 groups was analyzed by one-way ANOVA and Duncan multi-range tests. Values that do not share the same superscript on the same line are significantly different between groups. p <0.05, a, b, and c are significantly different between the groups.

CONCLUSIONS Mice fed HFD + 0.5% rucinol showed a significant reduction in body weight compared to the rucinol blend. This is an unexpected finding and cannot be predicted by those skilled in the art.

From the above examples, it is clear that garcinol results in an inhibition of adipogenesis and promotes weight loss in a dose-dependent manner as compared to a garcinol blend containing pterostilbene and anthocyanins. Garcinol also modifies the gut microbiota and increases the viable colonies of beneficial microorganisms-Akkermansia muciniphila, thereby maintaining and improving overall health and comfort. The present invention confirms that garcinol is an effective anti-obesity molecule and can be effectively administered alone or in combination with other weight-loss components for the management of obesity and related disorders. ..
Although the present invention has been described with respect to preferred embodiments, it should be clearly understood by those skilled in the art that the invention is not limited thereto. Rather, the scope of the invention should be construed only in relation to the appended claims.

Claims (17)

A method for the therapeutic management of obesity in mammals, wherein an effective concentration of a garcinol-containing composition is administered to the mammal to result in a) inhibition of adipogenesis, b) reduction of body weight and visceral fat in the mammal. A method, including steps. The method of claim 1, wherein the inhibition of adipogenesis is brought about by downregulation of a gene selected from the group consisting of PPARγ, cEBPα, FAS, AP2, resistin, and leptin. The method of claim 1, wherein the inhibition of lipogenesis is brought about by upregulation of a gene selected from the group consisting of p-AMPK, AMPK, and PREF-1. The method of claim 1, wherein the visceral fat is selected from the group consisting of mesenteric fat, peritoneal fat, and peri-gonadal fat. A method of achieving energy balance in a mammalian adipocyte system, wherein an effective amount of a garcinol-containing composition targeted to premammalian adipocytes and / or adipocytes is administered (a) to increase inhibition of adipocyte production. To specifically recruit effects and (b) brown adipocytes or brown-like (beige or bright) adipocytes and (c) induce a brown-like phenotype (beige or bright adipocytes) at the white adipocyte storage site. A method comprising achieving the effect of increasing the expression of adipocytes that function individually or in combination to bring about the effects of adipocyte utilization and energy balance in said mammals. 5. The method of claim 5, wherein the secretory factor comprises mitochondria UCP-1, PRDM16, PGC-1α, and BMP7. A method of modifying the intestinal microbial flora in a mammal, comprising the step of administering to the mammal an effective amount of a garcinol-containing composition to result in a change in the intestinal microbial flora. The gut microbiota is selected from Deferribacters, Pseudomonadota, Bacteroidotas, Verrucomicrobiota, and Firmicutes7. The method described in. The intestinal microbial flora includes the genus Lactobacillus, the genus Bacteroides, the genus Clostridium, the genus Anaerobranca, the genus Disgonomonas, the genus Dysgonomonas, and the genus Johnsononas. The method according to claim 7, wherein the method is selected from the genera Bacteroides, Oscillosspira, Parabacterroides, Akkermanisa, and Blautia. The intestinal microflora is Parabacteroides goldsteini, Bacteroides caccae, Johnsonella Ignava, Brutia winelaxi・ハンセンニ（Ｂｌａｕｔｉａ ｈａｎｓｅｎｎｉ）、アナエロブランカ・ザバルチンニ（Ａｎａｅｒｏｂｒａｎｃａ ｚａｖａｒｚｉｎｎｉ）、オシロスピラ・エアエ（Ｏｓｃｉｌｌｏｓｐｉｒａ ｅａｅ）、ムシスピリルム・シェドレリー（Ｍｕｃｉｓｐｉｒｉｌｌｕｓ ｓｃｈａｅｄｌｅｒｉ）、ブラウティア・コッコイデス（Ｂｌａｕｔｉａ ｃｏｃｃｏｉｄｅｓ）、アナエロツルンカス・コリホミニス（Ａｎａｅｒｏｔｒｕｎｃｕｓ ｃｏｌｉｈｏｍｉｎｉｓ）、ブチリビブリオ・プロテオクラスティカス（Ｂｕｔｙｒｉｖｉｂｒｏ ｐｒｏｔｅｏｃｌａｓｔｉｃｕｓ）、アッカーマンシア・ムシニフィラ（Ａｋｋｅｒｍａｎｓｉａ ｍｕｃｉｎｉｐｈｉｌａ）、ラクノスピラ・ペクチノスチザ（Ｌａｃｈｎｏｓｐｏｒａ ｐｅｃｔｉｎｏｓｃｈｉｚａ）、ペドバクター・クワンジャンゲンシス（Ｐｅｄｏｂａｃｔｅｒ ｋｗａｎｇｙａｎｇｅｎｓｉｓ）、アルカリフィルス・クロトンアトキシダンス（Ａｌｋａｌｉｐｈｉｌｕｓ ｃｒｏｔｏｎａｔｏｘｉｄａｎｓ） 、ラクトバチルス・サリバリウス（ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｌｉｖａｒｉｕｓ）、アナエロビブリオ・リポリティカス（Ａｎａｅｒｉｖｉｂｒｉａ ｌｉｐｏｌｙｔｉｃｕｓ）、ロドサーマス・クラルス（Ｒｈｏｄｏｔｈｅｒｍｕｓ ｃｌａｒｕｓ）、バクテロイデス・ステルコリロソリス（Ｂａｃｔｅｒｏｉｄｅｓ ｓｔｅｒｃｏｒｉｒｏｓｏｒｉｓ）、ルミノコッカス・フラベファシエンス（Ｒｕｍｉｎｏｃｏｃｏｃｃｕｓ ｆｌａｖｅｆａｃｉｅｎｓ）、バクテロイデス-Bacteroides xylanisolvens, Ruminococcus gnavus, Clostridium termitidis, Crossトリジウム・アルカリセルロシー（Ｃｌｏｓｔｒｉｄｉｕｍ ａｌｋａｌｉｃｅｌｌｕｌｏｓｉ）、エムチシシア・オリゴラフィカ（Ｅｍｔｉｃｉｃｉａ ｏｌｉｇｏｒａｐｈｉｃａ）、シュードブチリビブリオ・キシラニボランス（Ｐｓｅｕｄｏｂｕｔｙｒｉｖｉｂｒｏ ｘｙｌａｎｉｖｏｒａｎｓ）、アクチノマイセス・ナツラエ（Ａｃｔｉｎｏｍｙｃｅｓ ｎａｔｕｒａｅ）、ペプトニフィラス・コキシー（Ｐｅｐｔｏｎｉｐｈｉｌｕｓ ｃｏｘｉｉ）、およびドリコスペルマThe method of claim 7, wherein the method is selected from the group consisting of Clostridium curvum. Modification of the intestinal microflora is effective in the treatment management of diseases selected from the group consisting of obesity, cardiovascular complications, inflammatory bowel disease, Crohn's disease, celiac disease, metabolic syndrome, liver disease, and neuropathy. The method according to claim 7. A method for increasing the viable count of Akkermansia muciniphila in the intestine of a mammal, comprising the step of administering to the mammal an effective amount of a garcinol-containing composition to result in an increase in the colonies of the bacterium. .. 12. The method of claim 12, wherein an increase in the number of colonies of Akkermansia muciniphila causes weight loss through the AMPK signaling pathway by causing endocannabinoid release. A method of treating and managing hyperlipidemia in mammals, the administration of an effective concentration of garcinol-containing composition to (i) reduce the amount of total blood cholesterol levels in the blood of said mammal; (ii). ) Decrease the concentration of low-density lipoprotein (LDL) and very low-density lipoprotein (VLDL); (iii) increase the concentration of high-density lipoprotein (HDL); and (iv) increase the concentration of serum triglyceride. A method that includes steps that have the effect of lowering. 14. The method of claim 14, wherein the medical cause of hyperlipidemia is obesity. Garcinol-containing composition for use as a prebiotic agent. The composition is formulated with pharmaceutically / nutritionally acceptable excipients, auxiliaries, diluents, or carriers, tablets, capsules, syrups, gummy, powders, suspensions, emulsions, chews. 16. The composition of claim 16, which is orally administered in the form of tablets, candy, and groceries.

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